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Dan Ma,1,* Yunhui Fang,2,* ZiWei Wang,1 Mosang Yu,1 Xin Xin Zhou1 1Department of Gastroenterology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou City, People’s Republic of China; 2State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou City, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xin Xin Zhou, Department of Gastroenterology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou City, 310003, People’s Republic of China, Email zhouxinxin@zju.edu.cnBackground and Aim: Antibiotic resistance of Helicobacter pylori is increasing worldwide, lowering its efficacy in eradication therapy and posing a serious threat to human health. This study evaluated H. pylori resistance to antibiotics in the southeast region of China and explored factors related to eradication failure guided by antimicrobial susceptibility testing (AST).Methods: In this retrospective study, patients who tested positive underwent gastroscopy, and H. pylori infection was confirmed by histological staining and H. pylori culture. We determined the rate of H. pylori antibiotic resistance, success rate of AST-guided eradication therapy, and risk factors associated with treatment failure.Results: Among the 210 enrolled patients, 188 (89.5%) had successful cultures, and 183 (87.1%) underwent AST. The most common antibiotic resistance was to metronidazole and clarithromycin (89.6%), followed by levofloxacin (68.3%), and amoxicillin (14.2%). Furazolidone (3.0%) and tetracycline (0.5%) showed relatively low resistance rates. There were no statistically significant differences in the rates of resistance to MET, LEV, or AMX between naive and non-naive patients. However, CLA resistance rates in non-naive patients were significantly higher than those in naive patients. The overall success rate of AST-guided therapy was high and showed no significant difference between first-line and rescue therapy. Sex, age, prior therapy, and proton pump inhibitors (PPIs) or potassium-competitive acid blockers (P-CABs) use were not significantly associated with an increased risk of eradication failure in AST-guided therapy.Keywords: Antibiotic resistance, Culture, Helicobacter pylori, Eradication therapy.

Jiamin Yu,1 Yongxia Chen,1 Xiaoyan Pan,1 Ji Chen,1 Zhenhua Mai,1 Yuanli Zhang,1 Xiaoyan Wang,2– 4 Gaosheng Zhou,5 Sayed Adam Bukhari,6 Daqing Ma,6,7 Liehua Deng1 1Department of Critical Care Medicine, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524000, People’s Republic of China; 2Doctoral Scientific Research Center, Lianjiang People’s Hospital, Zhanjiang, 524400, People’s Republic of China; 3Affiliated Lianjiang People’s Hospital, Guangdong Medical University, Zhanjiang, 524400, People’s Republic of China; 4Laboratory of Southern Marine Science and Engineering, Zhanjiang, 524023, People’s Republic of China; 5Department of Critical Care Medicine, The First College of Clinical Medical Science, China Three Gorges University, Yichang Central People’s Hospital, Yichang, Hubei, 443003, People’s Republic of China; 6Division of Anaesthetics, Pain Medicine & Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea and Westminster Hospital, London, SW10 9NH, UK; 7Perioperative and Systems Medicine Laboratory, The Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, People’s Republic of ChinaCorrespondence: Liehua Deng, Email glinson@126.comPurpose: The early diagnosis of septic cardiomyopathy remains a challenge. The present work aims to evaluate the diagnostic and prognostic value of plasma soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) levels in septic cardiomyopathy when compared with traditional myocardial biomarkers.Methods: In the 143 sepsis enrolled patients, 67 and 76 patients were classified as non-septic cardiomyopathy and septic cardiomyopathy, respectively. Their blood samples were harvested up to 14th day after hospital admission for measurements of sTREM-1 and other biomarkers, such as N-terminal pronatriuretic peptide (NT-proBNP), highly sensitive troponin (TNT-HS), myoglobin (MYO), creatine kinase isoenzyme (CK-MB), etc. All the data were collected at 8:00 a.m. The area under the receiver operating characteristic curve was obtained to assess the diagnostic accuracy of those biomarkers. The Log rank test was utilized to evaluate the prognostic value of sTREM-1 on septic cardiomyopathy.Results: Circulating sTREM-1 showed a high specificity (88.1%) and moderate sensitivity (64.5%) to distinguish patients with septic cardiomyopathy in the 143 septic patients. The diagnostic efficiency of sTREM-1 was higher than inflammatory biomarkers and traditional myocardial markers. Logistic regression revealed that plasma sTREM-1 was an independent predictor of septic cardiomyopathy. Furthermore, in the whole septic cardiomyopathy cohorts, the sTREM-1 levels in the non-survivors were significantly higher than those of survivors during ICU stay. In addition, the left ventricular systolic dysfunction had a high odds ratio (3.968) to predict 90-day mortality in septic patients with cardiomyopathy.Conclusion: High plasma sTREM-1 level may be a diagnostic marker in predicting ICU poor outcome of patients with septic cardiomyopathy.Keywords: sepsis, septic cardiomyopathy, diagnosis, sTREM-1

Yifei Lu,1,* Minhua Ye,2,* Dehua Ma,2 Yu Chen2 1Department of Cardiology, Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, Linhai, People’s Republic of China; 2Department of Thoracic Surgery, Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University, Linhai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu Chen; Dehua Ma, Email chenyu5835@126.com; madh@enzemed.comObjective: To explore the association between survival status and the number of resected lymph nodes in node-negative esophageal squamous cell carcinoma(ESCC) after surgical treatment.Methods: This was a retrospective observational cohort study and data were obtained from the Surveillance, Epidemiology, and End Results program (SEER) and TaiZhou hospital in China. The data for subjects with negative lymph nodes and no distant metastasis (pN0M0) after post-operative pathology were screened. The nonlinear relationship between resected lymph node number and survival status in node-negative ESCC was conducted using restricted cubic spline regression analysis. The association between the number of resected lymph nodes and survival status in node-negative ESCC was evaluated by Cox proportional hazards regression models. Subgroup analysis based on different subgroups was also performed.Results: A total of 999 subjects were included in the study. Restricted cubic spline regression was used to show a U shaped association between the number of resected lymph nodes and survival status in node-negative ESCC, with low count associated with a decreased survival. To elucidate the association, we adjusted for age, sex, race, T stage, TNM (tumor node metastasis classification), location, grade, chemotherapy, and radiotherapy. As the resected lymph node number increased by one node, the survival status was improved by 2% (Hazard ratio(HR) = 0.98, 95% confidence interval (CI) 0.98– 0.99). Sensitivity analysis indicate that the effect size and direction in different subgroups are consistent, the results is stability in SEER.Conclusion: A low count of resected lymph nodes correlated with reduced survival in patients with ESCC, where resecting 25 to 28 or more nodes is considered optimal. Larger prospective studies are warranted to confirm these findings.Keywords: number of lymph node resection, survival status, cohort study, nonlinear relationship, node-negative esophageal squamous cell carcinoma

Xingtong Chen,1,&ast; Qian Chen,1– 3,&ast; Zhigang Qin,1,&ast; Azeem Alam,3 Hailin Zhao,3 Raha West,3 Xianzhe Liu,1 Jieyu Li,1 Xin Li,1 Bin Yi,1 Daqing Ma,2,3 Jianteng Gu1 1Department of Anesthesiology, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 2Perioperative and Systems Medicine Laboratory, National Clinical Research Center for Child Health, Children’s Hospital, Zhejiang University School of Medicine, Zhejiang, People’s Republic of China; 3Division of Anaesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, London, UK&ast;These authors contributed equally to this workCorrespondence: Jianteng Gu, Department of Anesthesiology, Southwest Hospital, Third Military Medical University (Army Medical University), 30 Gaotanyan Road, Chongqing, 400038, People’s Republic of China, Tel +86 23 68766122, Fax +86 23 65463270, Email jiantenggu@hotmail.com Daqing Ma, Division of Anaesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, London, UK, Email d.ma@imperial.ac.ukBackground: Dexmedetomidine (Dex) may have anti-inflammatory properties and potentially reduce the incidence of postoperative organ injury.Objective: To investigate whether Dex protects pulmonary and renal function via its anti-inflammatory effects in elderly patients undergoing prolonged major hepatobiliary and pancreatic surgery.Design and Setting: Between October 2019 and December 2020, this randomized controlled trial was carried out at a tertiary hospital in Chongqing, China.Patients: 86 patients aged 60– 75 who underwent long-duration (> 4 hrs) hepatobiliary and pancreatic surgery without significant comorbidities were enrolled and randomly assigned into two groups at a 1:1 ratio.Interventions: Patients were given either Dex or an equivalent volume of 0.9% saline (Placebo) with a loading dose of 1 μg kg− 1 for 10 min, followed by 0.5 μg kg− 1 hr− 1 for maintenance until the end of surgery.Main Outcome Measures: The changes in serum concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) were primary outcomes.Results: At one hour postoperatively, serum IL-6 displayed a nine-fold increase (P< 0.05) in the Placebo group. Administration of Dex decreased IL-6 to 278.09 ± 45.43 pg/mL (95% CI: 187.75 to 368.43) compared to the Placebo group (P=0.019; 432.16 ± 45.43 pg/mL, 95% CI: 341.82 to 522.50). However, no significant differences in TNF-α were observed between the two groups. The incidence of postoperative acute kidney injury was twice as high in the Placebo group (9.30%) compared to the Dex group (4.65%), and the incidence of postoperative acute lung injury was 23.26% in the Dex group, lower than that in the Placebo group (30.23%), although there was no statistical significance between the two groups.Conclusion: Dex administration in elderly patients undergoing major hepatobiliary and pancreatic surgery reduces inflammation and potentially protects kidneys and lungs.Registration: Chinese Clinical Trials Registry, identifier: ChiCTR1900024162, on 28 June 2019.Keywords: dexmedetomidine, elderly patients, long-duration hepatobiliary and pancreatic laparotomy, perioperative renal function, perioperative pulmonary function

Jun Shao,1 Xiya Wang,1 Haiyuan Yu,1 Wei Ding,1 Bin Xu,1 Dongsheng Ma,1 Xuechun Huang,2 Hongqing Yin1 1Department of Medical Ultrasound, Affiliated Kunshan Hospital of Jiangsu University, Kunshan, 215300, People’s Republic of China; 2Department of Medical Ultrasound, Affiliated Changzhou No. 2 People’s Hospital of Nanjing Medical University, Changzhou, 213000, People’s Republic of ChinaCorrespondence: Xuechun Huang; Hongqing Yin, Email ylgc20162022@163.com; sjwwy163@163.comBackground: The advantages of the dissecting the metastatic lymph nodes posterior to the right recurrent laryngeal nerve (LN-prRLN) remain a great deal of controversies in papillary thyroid carcinoma (PTC) patients without clinical evidence. The purpose of our retrospective research was to investigate the predictive factors of the LN-prRLN in cN0 PTC patients.Methods and Materials: Altogether 251 consecutive cN0 PTC participants accepted unilateral or bilateral thyroidectomy accompanied with LN-prRLN dissection between June 2020 and May 2023 were included in the research. Then, univariate and multivariate logical regression analysis were conducted to analyze the relationship between the LN-prRLN and these predictive factors, and a predictive model was also developed. Surgical complications of LN-prRLN dissection were also presented.Results: The rate of LN-prRLN was 17.9% (45/251) in cN0 PTC patients after the analysis of postoperative histology. The age < 55 years, multifocality, microcalcification, and BRAFV600E mutation were identified to be predictive factors of LN-prRLN in cN0 PTC patients. The risk score for LN-prRLN was calculated: risk score = 1.192 × (if age < 55 years) + 0.808 × (if multifocality) + 1.196 × (if microcalcification in nodule) + 0.918 × (if BRAFV600E mutation in nodule). The rates of the transient hypoparathyroidism and hoarseness were 1.2% (3/251) and 2.0% (5/251), respectively.Conclusion: The age < 55 years, multifocality, microcalcification, and BRAFV600E mutation are independent predictors of the LN-prRLN in cN0 PTC patients. An effective predictive model was established for predicting the LN-prRLN in cN0 PTC patients, with the aim to better guide the surgical treatment of PTC. A thorough inspection of the lateral compartment is recommended in PTC patients with risk factors. The multicenter research with long-term follow-up should be carried out to ascertain the optimal surgical approach for patients with PTC.Keywords: papillary thyroid carcinoma, ultrasound, BRAFV600E mutation, lymph nodes posterior to the right recurrent laryngeal nerve, predictive model

Danxu Ma,1 Abulaihaiti Maimaitimin,2 Yun Wang3 1Department of Anesthesiology and Pain Medicine, Beijing Chaoyang Hospital, Capital Medical University, Beijing, People’s Republic of China; 2Department of Rheumatology and Immunology, Moyu Uighur Medicine Hospital, Xinjiang, People’s Republic of China; 3Department of Anesthesiology, Beijing Friendship Hospital, Capital Medical University, Beijing, People’s Republic of ChinaCorrespondence: Yun Wang, Department of Anesthesiology, Beijing Friendship Hospital, Capital Medical University, No. 95 Yongan Road, Xicheng District, Beijing, 100050, People’s Republic of China, Email wangyun129@ccmu.edu.cnObjective: Cervicogenic headache (CEH) is a condition resulting from upper cervical spine dysfunction and associated structural and soft tissue abnormalities, significantly impacting patients’ quality of life. To acquire better therapeutic results, we presented a novel ultrasound-guided “three in one” approach plus interfascial plane (IFP) blocks for the treatment of CEH. This approach allows for the modulation of C2 dorsal root ganglion (DRG), third occipital nerve (TON), and C3 medial branch with one-point puncture. Additionally, it allows for IFP blocks between the upper neck and occipital muscles within the same scanning plane.Patients and Methods: We evaluated patients diagnosed with CEH from July 2021 to December 2022 in our pain clinic. We included those who did not respond to conservative treatment and single occipital nerve block, therefore received nerve block or pulsed radiofrequency (PRF) using the “Three in One” approach plus IFP blocks. The accuracy of the ultrasound-guided C2 DRG puncture procedures was confirmed through fluoroscopy with C-arm and the sensory testing of PRF. The therapeutic effect of these interventions was assessed using the numerical rating scale (NRS) scores during telephone follow-ups at 1, 3, and 6 months.Results: Utilizing the “Three in One” approach, a total of 5 patients diagnosed with CEH underwent nerve block plus IFP blocks, while 2 patients underwent PRF plus IFP blocks. Employing ultrasound-guided C2 DRG puncture procedures, the needle tip’s correct placement was confirmed through both fluoroscopy and sensory testing of PRF. Notably, none of the cases experienced any complications associated with the approach. Subsequent follow-up assessments revealed an improvement in the NRS scores for CEH in all patients.Conclusion: The ultrasound-guided “Three in One” approach plus IFP blocks may be a potential effective method for the treatment of CEH.Keywords: cervicogenic headache, interfacial plane blocks, pulsed radiofrequency, C2 dorsal root ganglion, third occipital nerve

Di Ma, Bingfen Li, Xiaoyan Liu, Ying Sun, Jingli Sun Department of Gynaecology and Obstetrics, General Hospital of the Northern Theater Command, Shenyang, People’s Republic of ChinaCorrespondence: Jingli Sun, Department of Gynaecology and Obstetrics, General Hospital of the Northern Theater Command, Shenyang, People’s Republic of China, Email zg3416@sina.comPurpose: We aimed to understand the current situation of smartphone addiction in pregnant women, and explored the moderating effect of self-efficacy between pregnancy stress and smartphone addiction.Patients and Methods: This study adopted a longitudinal design to collect pregnant women from the obstetrics and gynecology department of a tertiary hospital in Shenyang in 2020 from early pregnancy (T1) to their late pregnancy (T2). A total of 342 questionnaires were collected, including the Smartphone Addiction Scale-Short Version (SAS-SV), the Pregnancy Pressure Scale (PPS), and the Chinese version of the General Self-Efficacy Scale (GSES). Multiple hierarchical regression and simple slope test were used to test the moderating effect of self-efficacy.Results: Smartphone addiction in T2 (44.74) was more sever than in T1 (33.11). The interaction item of T1 pregnancy stress and T2 self-efficacy was positively correlated with smartphone addiction (β=0.205, P< 0.01) and explained an additional 3.2% variance (ΔR2=0.032, P< 0.01). The influence of pregnancy stress on smartphone addiction was gradually decreased in the low, mean, and high groups of self-efficacy.Conclusion: Smartphone addiction in late pregnancy was more severe than that in early pregnancy, possibly due to increased pregnancy stress. The self-efficacy of pregnant women could reduce the impact of pregnancy stress on smartphone addiction. Medical staff can alleviate the bad behavior by improving their self-efficacy.Keywords: late pregnancy, pregnancy stress, smartphone addiction, self-efficacy, moderating effect

Tongguo Miao,1 Xianzhe Lou,2 Shiming Dong,1 Xiaoxiao Zhang,1 Weiwei Guan,1 Ying Zhang,1 Lu Li,1 Xiwei Yuan,1 Dong Ma,2,&ast; Yuemin Nan1,&ast; 1Department of Traditional and Western Medical Hepatology, Hebei Medical University Third Hospital & Hebei International Joint Research Center for Liver Cancer Molecular Diagnosis, Hebei International Science and Technology Cooperation Base, Shijiazhuang, Hebei Province, 050051, People’s Republic of China; 2Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, Hebei Medical University, Shijiazhuang, Hebei Province, 050017, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuemin Nan, Department of Traditional and Western Medical Hepatology, Hebei Medical University Third Hospital & Hebei International Joint Research Center for Liver Cancer Molecular Diagnosis, Hebei International Science and Technology Cooperation Base, No. 139, Ziqiang Road, Shijiazhuang, Hebei Province, 050051, People’s Republic of China, Tel +86 18533112266, Email nanyuemin@163.com Dong Ma, Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, Hebei Medical University, No. 361, Zhongshan East Road, Shijiazhuang, Hebei Province, 050017, People’s Republic of China, Tel +86 15081650349, Email madong119@hebmu.edu.cnPurpose: The incidence of non-B and non-C hepatocellular carcinoma (NBNC-HCC) is increasing globally. Metabolically associated fatty liver disease (MAFLD) has been a contributing factor to this rising trend in NBNC-HCC incidence. The monocyte-to-high-density lipoprotein-cholesterol ratio (MHR) is a new prognostic marker that connects systemic inflammation with disorders of lipid metabolism. Therefore, MHR may be a potential prognostic predictor of patients with MAFLD-related HCC (MAFLD-HCC). This study aims to investigate the relationship between the MHR and prognosis of patients with MAFLD-HCC and construct a novel prognostic prediction tool for MAFLD-HCC.Patients and Methods: This retrospective study of patients with MAFLD-HCC included training (n = 112) and internal validation (n = 37) cohorts. Univariate and multivariate Cox proportional hazard regression analysis was conducted to identify independent risk factors of survival. A visual nomogram was constructed to assess the performance of the two groups. Furthermore, receiver operating characteristic (ROC) curves and calibration curves were used to verify the prognostic discriminative ability of this nomogram, even in the MHR, ALBI grade, and MHR–ALBI model.Results: Univariate and multivariate analyses revealed that extrahepatic metastases, Vascular invasion, Barcelona staging B, C, D, elevated ALBI Grade 3, C-reactive protein (CRP), and MHR were independent risk factors for the prognosis of MAFLD-HCC. Moreover, calibration plots showed good discrimination and consistency when the significant factors were entered into the nomogram. Meanwhile, the MHR strongly correlated with the prognosis of cancer under a background of MAFLD-HCC, with a sensitivity of 88.89% and a specificity of 79.61%. Importantly, the performance of the MHR alone (AUC = 86.2) was not only superior to the ALBI grade (AUC = 63.8) but was comparable to the combination of MHR and ALBI (AUC = 88.5).Conclusion: The novel nomogram demonstrated good value in predicting the overall survival of patients with MAFLD-HCC. The MHR may be a potential predictor of prognosis.Keywords: lipid metabolism, MAFLD-HCC, prognostic prediction, nomogram, MHR

Ning Zhang,1,&ast; Tong Liu,1,&ast; Jianan Wang,2,&ast; Yingying Xiao,1 Ying Zhang,1 Jun Dai,1 Zhihong Ma,1,3 Donglai Ma4 1School of Basic Medicine, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 2Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 3Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns, Shijiazhuang, Hebei, 050200, People’s Republic of China; 4School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhihong Ma; Donglai Ma, Hebei University of Chinese Medicine, Xingyuan Road 3, Shijiazhuang, Hebei, 050200, People’s Republic of China, Tel +86 31189926238, Email mazh1969@163.com; mdl_hebei@aliyun.comBackground: Metabolic associated fatty liver disease (MAFLD) is a chronic disease characterized by excessive lipid deposition in the liver without alcohol or other clear liver-damaging factors. AMP-activated protein kinase (AMPK)/silencing information regulator (SIRT)1 signaling pathway plays an important role in MAFLD development. Si-Ni-San (SNS), a traditional Chinese medicine, has shown reducing hepatic lipid deposition in MAFLD rats, however, the underlying mechanisms of SNS are barely understood.Purpose: The aim of this research was to investigate the mechanisms of SNS in reducing hepatic lipid deposition in MAFLD rats by regulating AMPK/SIRT1 signaling pathways.Methods: The components of SNS were determined by high performance liquid chromatography with mass spectrometry (HPLC-MS) analysis. MAFLD rats were induced by high-fat and high-cholesterol diet (HFHCD), and treated by SNS. SNS-containing serum and Compound C (AMPK inhibitor) were used to treat palmitic acid (PA)-induced HepG2 cells. To elucidate the potential mechanism, lipid synthesis-related proteins (SREBP-1c and FAS), fatty acid oxidation (PPARα and CPT-1), and AMPK/SIRT1 signaling pathway (p-AMPK and SIRT1) were assessed by Western blot.Results: SNS improved serum lipid levels, liver function and reduced hepatic lipid deposition in MAFLD rats. SNS-containing serum reduced lipid deposition in PA-induced HepG2 cells. SNS could up-regulate protein expressions of PPARα, CPT-1, p-AMPK and SIRT1, and down-regulate protein expressions of SREBP-1c and FAS. Similar effects of SNS-containing serum were observed in PA-induced HepG2 cells. Meanwhile, Compound C weakened the therapeutic effects of SNS-containing serum on lipid deposition.Conclusion: SNS could reduce hepatic lipid deposition by inhibiting lipid synthesis and promoting fatty acid oxidation, which might be related with activating the AMPK/SIRT1 signaling pathway. This study could provide a theoretical basis for the clinical use of SNS to treat MAFLD.Graphical Abstract: Keywords: Si-Ni-San, MAFLD, lipid deposition, lipid synthesis, fatty acid oxidation, AMPK

Yunxiao Wu,1 Yan Wang,2,3 Changming Wang,4 Fujun Zhao,5 Dandi Ma,6 Zhifei Xu,6,7 Xin Ni5,7 1Beijing Key Laboratory of Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, People’s Republic of China; 2Center on Aging Psychology, CAS Key Laboratory of Mental Health, Institute of Psychology, Beijing, People’s Republic of China; 3Department of Psychology, University of Chinese Academy of Sciences, Beijing, People’s Republic of China; 4Department of Neurosurgery, Xuanwu Hospital, Capital Medical University, Beijing, People’s Republic of China; 5Department of Otolaryngology, Head and Neck Surgery, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, People’s Republic of China; 6Respiratory Department 1, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, People’s Republic of China; 7MOE Key Laboratory of Major Diseases in Children, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, People’s Republic of ChinaCorrespondence: Zhifei Xu, Respiratory Department 1, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, 56 Nanlishi Road, Xicheng, Beijing, 100045, People’s Republic of China, Tel +8613521660077, Email zhifeixu@aliyun.com Xin Ni, Department of Otolaryngology, Head and Neck Surgery, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, 56 Nanlishi Road, Xicheng, Beijing, 100045, People’s Republic of China, Tel +8613370115099, Email nixin@bch.com.cnPurpose: To explore the characteristics of the attentional network and related factors in children with sleep-disordered breathing (SDB).Patients and Methods: A total 228 children (200 children aged 6– 10 years with snoring or mouth breathing, admitted to our hospital from May 2020 to July 2022, and 28 healthy children recruited from the community as the control group) were enrolled. All participants underwent polysomnography (PSG) and completed the ADHD rating scale and child version of the Attention Network Test. According to their SDB history and obstructive apnea hypopnea index (OAHI), the participants were divided into control (n = 28), primary snoring (PS; n = 67) and obstructive sleep apnea (OSA; n = 133) groups.Results: The OSA and PS groups were younger than controls (P < 0.05). The proportion of boys was higher in the OSA than control group (P < 0.05). Body mass index was higher in the OSA than control and PS groups (P < 0.01). Attention deficit and hyperactive impulsivity scores were independently associated with the OAHI (P < 0.001). The efficiency of the alerting network was higher in the OSA than in controls (P = 0.020), but was not correlated with OAHI after adjusting for age, sex and SDB history duration (P > 0.05).Conclusion: Children with OSA have impaired attention, characterized by excessive alerting network activation. However, alerting network efficiency did not change linearly with disease severity. More research is needed to elucidate the neural mechanisms underlying attention deficits in pediatric OSA.Keywords: child, obstructive sleep apnea, attention, alerting network

Hao Wang, Jiwei Zhang, Dongyue Ma, Ziwei Zhao Department of Andrology, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, People’s Republic of ChinaCorrespondence: Jiwei Zhang, Department of Andrology, Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing, 100091, Email zhangjiweixyyy@163.comAbstract: Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is one of the most common diseases in urology, and its prevalence has been increasing, causing serious urogenital problems in men. Several targeted oral drug therapies exist to treat the pelvic pain experienced in this disease. However, these therapies may have potential adverse effects, and due to the unique location of the prostate, delivery of medications to the target lesion is difficult. Consequently, many patients seek alternative therapies. Acupuncture is a well-established treatment method in traditional Chinese medicine that can improve CP/CPPS symptoms and reduce pain. However, previous systematic reviews overlooked the significance of the characteristics and mechanisms of acupuncture therapy. This study aimed to summarize the characteristics and benefits of acupuncture therapy for CP/CPPS based on relevant literature and predict and analyze its related mechanisms.Keywords: acupuncture, chronic prostatitis, chronic pelvic pain syndrome, mechanism, traditional Chinese medicine, pain

Diming Wang,1,&ast; Wei Ye,2,&ast; Dongmei Chen,1 Qingming Shi,1 Dongchun Ma3 1Department of Oncology, Anhui Chest Hospital, Hefei, 230022, People’s Republic of China; 2Department of Pathology, Anhui Chest Hospital, Hefei, 230022, People’s Republic of China; 3Department of Thoracic Surgery, Anhui Chest Hospital, Hefei, 230022, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Dongchun Ma, Department of Thoracic Surgery, Anhui Chest Hospital, Hefei, 230022, People’s Republic of China, Email madongchun126@126.com Qingming Shi, Department of Oncology, Anhui Chest Hospital, Hefei, 230022, People’s Republic of China, Email shqm0324@163.comAbstract: The transformation of lung adenocarcinoma to small cell lung cancer (SCLC) following treatment with epidermal growth factor (EGFR) receptor tyrosine kinase inhibitors (TKIs) is a relatively common phenomenon. However, transformation of non-small cell lung cancer (NSCLC) to SCLC following treatment with immunotherapy is very rare. Here, we report a case of a 56-year-old patient diagnosed with driver gene mutation-negative lung squamous cell carcinoma (SCC). He received four cycles of immunotherapy with sugemalimab and chemotherapy with albumin paclitaxel in combination with carboplatin, and a partial response was achieved. Subsequently, the patient received 5 cycles of immunotherapy with sugemalimab. However, he developed rapid progression of mediastinal lymph nodes, and biopsy results showed transformation to SCLC. His tumor did not respond to the next line of carboplatin combined with etoposide, and he died six months after the discovery of SCLC transformation. In conclusion, SCLC transformation is also an important resistance mechanism for lung SCC patients treated with immunotherapy and predicts a very poor outcome. Repeat biopsy is needed for advanced lung SCC that has progressed with immunotherapy.Keywords: lung squamous cell carcinoma, immunotherapy, small cell transformation

Dengming Lai,1,&ast; Kun Zhu,2,&ast; Sisi Li,3,&ast; Yi Xiao,1 Qi Xu,4 Yisheng Sun,5 Pingping Yao,5 Daqing Ma,6,7 Qiang Shu7 1Department of Neonatal Surgery, Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, People’s Republic of China; 2Department of Pathology, The Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, People’s Republic of China; 3School of Medicine, Hangzhou City University, Hangzhou, People’s Republic of China; 4School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, People’s Republic of China; 5Key Laboratory of Vaccine, Prevention and Control of Infectious Disease of Zhejiang Province, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, People’s Republic of China; 6Division of Anaesthetics, Pain Medicine & Intensive Care, Department of Surgery & Cancer, Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, London, UK; 7Department of Thoracic and Cardiovascular Surgery, Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qiang Shu, Department of Thoracic and Cardiovascular Surgery, Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310052, People’s Republic of China, Tel +86-13906500193, Email shuqiang@zju.edu.cn Dengming Lai, Department of Neonatal Surgery, Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310052, People’s Republic of China, Tel +86-13567164728, Email dengming_lai@zju.edu.cnBackground: SARS-CoV-2-induced acute lung injury but its nucleocapsid (N) and/or Spike (S) protein involvements in the disease pathology remain elusive.Methods: In vitro, the cultured THP-1 macrophages were stimulated with alive SARS-CoV-2 virus at different loading dose, N protein or S protein with/without TICAM2-siRNA, TIRAP-siRNA or MyD88-siRNA. The TICAM2, TIRAP and MyD88 expression in the THP-1 cells after N protein stimulation were determined. In vivo, naïve mice or mice with depletion macrophages were injected with N protein or dead SARS-CoV-2. The macrophages in the lung were analyzed with flow cytometry, and lung sections were stained with H&E or immunohistochemistry. Culture supernatants and serum were harvested for cytokines measurements with cytometric bead array.Results: Alive SARS-CoV-2 virus or N protein but not S protein induced high cytokine releases from macrophages in a time or virus loading dependent manner. MyD88 and TIRAP but not TICAM2 were highly involved in macrophage activation triggered by N protein whilst both inhibited with siRNA decreased inflammatory responses. Moreover, N protein and dead SARS-CoV-2 caused systemic inflammation, macrophage accumulation and acute lung injury in mice. Macrophage depletion in mice decreased cytokines in response to N protein.Conclusion: SARS-CoV-2 and its N protein but not S protein induced acute lung injury and systemic inflammation, which was closely related to macrophage activation, infiltration and release cytokines.Keywords: SARS-CoV-2, S protein, N protein, macrophage, cytokine, acute lung injury

Cheng Ma,1– 4,&ast; Yin Cao,1,&ast; Guang Zhang,1– 3,&ast; Jiannan Qiu,2 Yan Zhou,1 Peng Wang,2 Shuo Wang,2 Dongliang Yan,1 Ding Ma,2 Chunping Jiang,1– 3 Zhongxia Wang1– 3 1Department of Hepatobiliary Surgery, Drum Tower Clinical College of Nanjing Medical University, Nanjing, People’s Republic of China; 2Department of Hepatobiliary Surgery, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, People’s Republic of China; 3Department of Tissue Engineering, Jinan Microecological Biomedicine Shandong Laboratory, Jinan, People’s Republic of China; 4Department of Gastrointestinal Surgery, Xuzhou Central Hospital, Xuzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhongxia Wang; Chunping Jiang, Email freud_t@126.com; chunpingjiang@163.comBackground: The prediction of prognosis of hepatocellular carcinoma (HCC) is of great significance in improving disease outcome and optimizing clinical management, while reliable prognostic indicators are lacking. This study was conducted to develop readily-to-use nomograms for prognosis prediction of HCC after hepatectomy.Materials and Methods: Data of eligible patients were collected and analyzed retrospectively. Independent prognostic factors were identified by Cox regression, and nomograms for the prediction of disease-free survival (DFS) and overall survival (OS) were developed. The performance of the nomograms was evaluated by receiver operating characteristics (ROC) curves, C-indexes and calibration curves and was verified by the validation cohort. The predictive value of the nomograms was also compared with the 8th edition of American Joint Committee on Cancer (AJCC) Tumor-Node-Metastasis (TNM) and the Barcelona Clinic Liver Cancer (BCLC) staging systems.Results: In total, 599 patients were enrolled in the analysis: 420 in the training cohort and 179 in the validation cohort. The optimal cut-off value of Gamma-Glutamyl Transpeptidase-to-Lymphocyte Ratio (GLR) was 19.5. GLR contributed significantly to the nomograms with good predictive power. In ROC analyses, the areas under curve (AUCs) of the nomograms for 1-, 3- and 5-year DFS and OS prediction were 0.758, 0.756, 0.734 and 0.810, 0.799, 0.758, respectively. The C-indexes of the DFS nomogram were 0.697 (95% CI 0.665– 0.729) in the training cohort and 0.710 (95% CI 0.664– 0.756) in the validation cohort. For OS prediction, the C-indexes were 0.741 (95% CI 0.704– 0.778) and 0.758 (95% CI 0.705– 0.811) in the training and validation cohorts, respectively. The calibration curves demonstrated satisfactory agreement between nomogram predictions and actual observations. The nomograms demonstrated superior predictive performance to the TNM and the BCLC staging systems.Conclusion: Our novel nomograms showed adequate performance in the prediction of HCC prognosis after hepatectomy, which may facilitate the risk stratification and individualized management of HCC patients.Keywords: hepatocellular carcinoma, nomogram, gamma-glutamyl transferase, lymphocyte, hepatectomy

Ruijuan Guo,1 Huanrong Qiu,1 Huili Li,2 Danxu Ma,2 Yun Guan,3 Yun Wang2 1Department of Anesthesiology, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050, People’s Republic of China; 2Department of Anesthesiology, Beijing Chaoyang Hospital, Capital Medical University, Beijing, 100020, People’s Republic of China; 3Department of Anesthesiology and Critical Care Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USACorrespondence: Yun Wang, Department of Anesthesiology, Beijing Chaoyang Hospital, Capital Medical University, No. 8, Gongtinan Road, Chaoyang District, Beijing, 100020, People’s Republic of China, Tel +86-010-85231330, Fax +86-10-65077808, Email wangyun129@ccmu.edu.cnPurpose: Subcutaneous infiltration of capsaicin, which initially activates transient receptor potential vanilloid 1 (TRPV1) receptors, can subsequently desensitize TRPV1-expressing nociceptors and induce analgesia in different pain models. Yet, whether the modulation of keratinocytes may also contribute to the analgesic action of capsaicin treatment remains unclear. In a rat model of postoperative pain, we tested the hypothesis that subcutaneous injection of capsaicin inhibited the proliferation of epidermal keratinocytes and their expression of pronociceptive inflammatory mediators after plantar incision.Methods: The plantar incision model was carried out in the current study. Behavioral tests were used to evaluate postoperative pain-related behaviors in rats. Immunohistochemistry was used to investigate epidermal keratinocytes proliferation and expression of pro-inflammatory mediators in keratinocytes in rats.Results: Behaviorally, plantar incision induced robust postoperative pain hypersensitivity. However, subcutaneous pretreatment of capsaicin (1%) but not the vehicle, prevented the development of postoperative pain. There was an increased proliferation of keratinocytes and the expressions of interleukin-1β (IL-1β) and tumour necrosis factor-alpha (TNF-α) in keratinocytes at 3 d and 7 d after plantar incision. However, these changes were also significantly attenuated by capsaicin pretreatment.Conclusion: Our findings suggest that capsaicin pretreatment may inhibit incision-induced keratinocytes proliferation and reduce their expression of pronociceptive inflammatory mediators under postoperative pain conditions, which represents a peripheral non-neuronal mechanism of capsaicin-induced analgesia.Keywords: capsaicin, keratinocyte, postoperative pain, inflammatory mediators

Hao Tian,1,&ast; Tingting Zhao,1,&ast; Yanling Li,1,&ast; Na Sun,1 Dandan Ma,1 Qiyun Shi,1 Guozhi Zhang,1 Qingqiu Chen,1 Kongyong Zhang,1 Ceshi Chen,2,&ast; Yi Zhang,1 Xiaowei Qi1 1Department of Breast and Thyroid Surgery, Southwest Hospital, Army Medical University, Chongqing, People’s Republic of China; 2Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiaowei Qi; Yi Zhang, Department of Breast and Thyroid Surgery, Southwest Hospital, Army Medical University, Gaotanyan Street 29, Chongqing, 400038, People’s Republic of China, Tel/Fax +86-23-68754160, Email qxw9908@foxmail.com; zhangyi1489@sina.comBackground: Epigenetic modification of chromatin is an important step in the regulation of gene expression. The chromobox family proteins (CBXs), as epigenetic modifier, may play a vital role in tumorigenesis and cancer progression. Herein we explored the correlation between CBXs and breast cancer (BC) via the bioinformatics approach and qRT-PCR validation.Methods: Several databases, including GEPIA, TCGA, GEO, K-M plotter, STRING, DAVID, cBioPortal, CIBERSORT, and HPA were employed to analyze the expression levels of CBXs and the correlations between CBXs and prognosis (overall and recurrence-free survival) in BC. We analyzed molecular functions, genetic variations, transcription factors of CBXs, and immune cell infiltration status. ROC curve analysis was performed to determine the predictive value of CBXs. RNA extracted from 11 human BC and paired adjacent normal tissues were subjected to qRT-PCR.Results: The mRNA expression level of CBX1– 5 was significantly upregulated, while that of CBX7 was significantly downregulated in BC; no expression disparities were observed in CBX6/8 expression. Further, high mRNA expression of CBX1/2/3/4/8 correlated with advanced BC, whereas high mRNA expression of CBX6/7 correlated with early BC. High mRNA expressions of CBX1/2/3/5 predict poor OS and RFS, while higher mRNA expressions of CBX6/7 predict better OS and RFS in patients with BC. ROC curve analysis revealed that CBX3 showed excellent discriminatory ability. Gene ontology enrichment analysis showed that CBXs primarily participated in SUMOylation and post-/transcriptional regulation. Moreover, they presented varying degrees of amplification in BC tissues and were related to the infiltration of various immune cells.Conclusion: CBXs can serve as putative biomarkers for BC. Further studies are warranted to determine the exact molecular mechanisms underlying the action of CBXs in BC, particularly CBX1/2/3/5/7.Keywords: chromobox family proteins, breast cancer, prognosis, bioinformatics, qRT-PCR

Fang Zhao,1 Deyong Ma,2 Tingting Wang,3 Yan Zhang,2 Ying Dong,4 Jian Zhao2 1Department of Obstetrics and Gynecology, Beijing Jishuitan Hospital, Beijing, 100035, People’s Republic of China; 2Department of Obstetrics and Gynecology, Peking University First Hospital of Obstetrics and Gynecology, Beijing, 100034, People’s Republic of China; 3Peking University Department of Medicine, Beijing, 100191, People’s Republic of China; 4Department of Pathology, Peking University First Hospital of Obstetrics and Gynecology, Beijing, 100034, People’s Republic of ChinaCorrespondence: Jian Zhao, Email JianZhao112@outlook.comObjective: The aim of this study was to clinically evaluate the application of liquid-based cytology P16, cytologic evaluation, and high-risk human papillomavirus (HR-HPV) testing in cervical cancer screening.Methods: This study screened 900 women, who attended the outpatient clinic, according to the exclusion criteria of study participants. The study participants’ screening results of liquid-based cytology P16, cytologic evaluation, and HR-HPV testing were analyzed. According to the pathological results of the biopsy, the efficacy of different screening strategies for the identification of high-grade lesions was evaluated.Results: The positive rate of p16 expression increased with the severity of cervical lesions. P16 had the highest sensitivity and negative predictive value in identifying high-grade lesions (98.45% and 99.67%, respectively). Liquid-based Papanicolaou test (LBP), on the other hand, had the lowest sensitivity (85.27%) but the highest specificity (85.88%). HR-HPV’s positive predictive value and accuracy rate were the lowest (32.77% and 70.03%, respectively). The difference was statistically significant (P < 0.05). Dual combinations of certain tests were set up for this study; P16+LBP, HPV+LBP, and P16+HPV had sensitivities of 98.45%, 96.90%, and 99.22%, and specificities of 80.29%, 63.42%, and 64.33%, respectively. The P16 screening rates of histological and liquid-based cytology approaches were 75.74%.Conclusion: Compared with traditional LBP+HPV, the application of a test that solely screen for P16 or the combined screening method that involves the screening of P16 is more effective in identifying high-grade lesions.Keywords: biomarkers, liquid-based cytology, p16, HR-HPV, screening of high-grade cervical lesions

Qian Chen,1,2,&ast; Zhigang Qin,1,&ast; Yibing Sun,3 Xiangfeng Liu,1 Aurelie Pac Soo,2 Enqiang Chang,2 Qizhe Sun,2 Bin Yi,1 Dong-Xin Wang,3 Hailin Zhao,2 Daqing Ma,2 Jianteng Gu1 1Department of Anaesthesiology, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 2Division of Anaesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, London, UK; 3Department of Anaesthesiology and Critical Care Medicine, Peking University First Hospital, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jianteng Gu, Department of Anaesthesiology, Southwest Hospital, Third Military Medical University, 30 Gaotanyan Road, Chongqing, People’s Republic of China, Tel +86 23 68765366, Fax +86 2365463270, Email jiantenggu@hotmail.com Daqing Ma, Division of Anaesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea and Westminster Hospital, London, UK, Tel +44 020 3315 8495, Fax +44 020 3315 5109, Email d.ma@imperial.ac.ukPurpose: This study aims to investigate the cytoprotective and anti-inflammatory effects of an α2-adrenoreceptor (α2-AR) agonist, dexmedetomidine (Dex), on lipopolysaccharides (LPS)-induced acute lung injury and underlying mechanisms with focus on alveolar macrophage polarization modulation.Methods: C57BL/6 mice were intraperitoneally injected LPS (10 mg/kg) with or without Dex (25 μg/kg) and/or α2-AR antagonist atipamezole (Atip, 500 μg/kg). Lung tissues were then analysed to determine injuries. In vitro, human pulmonary epithelial cells (A549) and mice alveolar macrophages (MH-S) were exposed to LPS (10 ng/mL) with or without different concentrations of Dex (0.1– 100 nM). Alveolar macrophage polarization, NLRP3 inflammasome activation and inflammatory responses were determined. PTEN/Akt signaling and its downstream transcriptional factors as targets for macrophage polarization were assessed.Results: Dex treatment significantly reduced pro-inflammatory M1 macrophage polarization and NLRP3 inflammasome activation in the lungs relative to the mice treated with LPS. The similar pattern reduction of NLRP3 inflammasome activation by Dex was also found in A549 cells. Atip partly reversed the anti-inflammatory effects of Dex. In cultured alveolar macrophages, Dex reduced LPS-mediated expression of IL-1, − 6 and TNF-α receptors while promoting alveolar macrophages differentiation towards a M2 anti-inflammatory phenotype. Additionally, LPS increased Akt signaling activation in a time-dependent manner, which was further activated by Dex via inhibiting phosphatase and tensin homolog (PTEN). The action of Dex on Akt signaling shifted alveolar macrophages from M1 to M2 phenotype through increasing STAT6 and IRF4 transcriptional factors.Conclusion: Dex protected against LPS-induced lung injury and suppressed LPS-induced pulmonary inflammatory responses by attenuating the NLRP3 inflammasome activation and promoting anti-inflammatory M2 macrophage polarization.Keywords: sepsis, acute lung injury, macrophage polarization, dexmedetomidine, Akt signaling

Yajing Fu,1 Dan Ma,2 Fengyan Fan,3 Tongke Sun,1 Ruiqi Han,1 Yanran Yang,1 Jun Zhang1 1Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China; 2Department of Orthodontics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, People’s Republic of China; 3Department of Orthodontics, Hangzhou Stomatological Hospital, Hangzhou, People’s Republic of ChinaCorrespondence: Jun Zhang, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, No. 44-1 Wenhua Road West, Jinan, People’s Republic of China, Tel +86 139 5310 9816, Email zhangj@sdu.edu.cnPurpose: Stem cells from the apical papilla (SCAPs) are promising seed cells for tissue regeneration medicine and possess the osteogenic differentiation potential. Wnt5a, a typical ligand of the noncanonical Wnt pathway, exhibits diverse roles in the regulation of osteogenesis. The transcriptional co-activator with PDZ-binding motif (TAZ, WWTR1) is a core regulator in the Hippo pathway and regulates stem behavior including osteogenic differentiation. This study aims to examine how Wnt5a regulates SCAPs osteogenesis and explore the precise mechanistic relationship between Wnt5a and TAZ.Methods: SCAPs were isolated from developing apical papilla tissue of extracted human immature third molars in vitro. ALP staining, ALP activity and Alizarin red staining were used to evaluate osteogenic capacity. Osteogenic-related factors were assessed by qRT-PCR or Western blotting. Additionally, the receptor tyrosine kinase-like orphan receptor 2 (ROR2) was detected by immunocytofluorescence staining and silenced by small interfering RNA to verify the function of Wnt5a/ROR2 in TAZ-mediated osteogenesis. And we constructed TAZ-overexpression and β-catenin-overexpression SCAPs generated by lentivirus to explore the precise mechanistic relationship between Wnt5a and TAZ.Results: Wnt5a (100ng/mL) significantly suppressed ALP activity, mineralization nodules formation, expression of osteogenic-related factors. Meanwhile, it decreased the expression of TAZ mRNA and protein. TAZ overexpression promoted osteogenesis of SCAPs while Wnt5a could block TAZ-mediated osteogenesis. Furthermore, ROR2 siRNA (siROR2) was found to upregulate TAZ and canonical Wnt pathway signaling related molecules such as β-catenin, GSK3β and p-GSK3β. The suppression of Wnt5a/ROR2 on osteogenesis was significantly reversed by β-catenin overexpression through Wnt5a/ROR2/β-catenin/TAZ pathway.Conclusion: Taken together, the present study demonstrates that Wnt5a suppresses TAZ-mediated osteogenesis of SCAPs and there may be a Wnt5a/ROR2/β-catenin/TAZ pathway regulating osteogenesis of SCAPs. Moreover, Wnt5a could be a candidate for regulators in tissue regeneration.Graphical Abstarct: Keywords: Wnt5a, TAZ, stem cells from the apical papilla, osteogenesis, β-catenin

Maoti Wei,1,&ast; Li Dong,2,&ast; Fenghua Wang,1 Kai Cui,3 Delong Ma,2 Ning Yang,4 Yuming Li4 1Center of Clinical Epidemiology, TEDA International Cardiovascular Hospital, Tianjin, People’s Republic of China; 2Department of Pediatrics, Center Hospital of Beikuo Town, Rizhao City, Shandong Province, People’s Republic of China; 3Department of Disease Prevention and Control, Hospital of Gaoxing Town, Rizhao City, Shandong Province, People’s Republic of China; 4Department of Hypertension, TEDA International Cardiovascular Hospital, Tianjin, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ning Yang; Yuming Li Email yangningzxdl@sina.com; cardiolab@live.comObjective: To understand the prevalence and control of type 2 diabetes mellitus (T2DM) in a rural area, a cross-sectional study was carried out.Methods: Blood glucose was determined in residents over 60 years during the National Basic Public Health Service project carrying on in a rural town of Shandong province. Combined with detail information of diabetic history, the prevalence rate of this disease was calculated with a cutoff of more than 7.0 mmol/L of fasting plasma glucose (FPG). Furthermore, according to FPG levels, the control rate of diabetes was evaluated. Basic information and clinical laboratory tests were analyzed with proper methods for univariate analysis. Multivariable logistic analysis was used to explore multiple variables associated with the control effect of T2DM.Results: The prevalence rate of T2DM was 13.5% (95% CI 12.4%∼ 14.7%) without age difference in residents over 60-year old (X2= 1.078, P = 0.898). And the rate was higher in the female (16.1%, 95% CI 14.4%∼ 17.8%) than that in the male (11.3%, 95% CI 9.7%∼ 12.9%). The control rate of diabetes was 44.5% (95% CI 39.4– 49.6%, cutoff < 6.1 mmol/L for FPG) and 59.6% (95% CI 54.5– 64.6%, cutoff < 7.0 mmol/L for FPG), respectively. Most T2DM residents had balance foods, and a high proportion of drugs or insulin were used; however, the proportion of keeping exercise and modifying behaviors was low. Nearly 80% of the diabetic individuals used at least one drug or insulin. However, the more drugs or insulin used, the higher the FPG level was.Conclusion: A relative high level of T2DM prevailed in the rural area under fast economy-transforming. T2DM was controlled poor and measures should be further optimized to achieve ideal effect, especially for the use of anti-diabetic drugs.Keywords: rural area, type 2 diabetes mellitus, the control of diabetes

Chunlei Wu, Denhua Ma, Yu Chen Department of Cardiac Surgery, Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, Linhai, 317000, People’s Republic of ChinaCorrespondence: Yu Chen People’s Republic of ChinaEmail chenyu5835@126.comPurpose: Here, we sought to determine the association between pulse pressure difference and the incidence of type 2 diabetes mellitus (T2DM) in Chinese people.Methods: This study involved 211,814 participants among whom 4156 had been diagnosed with T2DM. The correlation between pulse pressure difference and T2DM incidence in Chinese people was determined by multivariate analysis. A smooth curve fitting diagram was then used to explore correlation between pulse pressure difference and T2DM incidence. Finally, the inflection point in the correlation between pulse pressure difference and the T2DM incidence was located by piecewise linear regression.Results: To understand the relationship, adjustments were made for sex, age, total serum cholesterol (TC), fasting blood glucose (FPG), triglyceride (TG), alanine aminotransferase (ALT), family history of diabetes, body mass index (BMI), blood urea nitrogen (BUN), drinking status, and smoking status. Diabetes incidence increased by 0.3% [HR 1.003 (1.001, 1.005), p = < 0.05] for every 1mmHg increase in pulse pressure difference. Smooth curve analysis showed that, when pulse pressure difference was ≤ 35mmHg, diabetes incidence negatively correlated to pulse pressure difference [HR 0.972 (0.953, 0.972) p = 0.053]. However, when pulse pressure difference was > 35mmHg, diabetes incidence increased with increasing pulse pressure difference [HR 1.044 (1.042, 1.047) p = < 0.001]. And between pulse pressure difference and fasting blood glucose in the final visit, the blood glucose level increased with the elevation of pulse pressure.Conclusion: The risk of diabetes was lowest at about 35mmHg pulse pressure difference.Keywords: pulse pressure difference, diabetes mellitus incidence, Chinese population, cohort study

Junshuai Xue,1,&ast; Delin Ma,2,&ast; Jianjun Jiang,1 Yang Liu1 1Department of General Surgery, Vascular Surgery, Shandong University Qilu Hospital, Jinan City, Shandong Province, People’s Republic of China; 2Department of General Surgery, Shandong University Qilu Hospital, Jinan City, Shandong Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yang LiuDepartment of General Surgery, Vascular Surgery, Shandong University Qilu Hospital, No. 107, Road Wen Hua Xi, Jinan City, Shandong Province, 250012, People’s Republic of ChinaTel +86 18560088317Email liuyang_sdu@126.comAbstract: Venous thromboembolism (VTE), which includes deep vein thrombosis (DVT) and pulmonary embolism (PE), has been an important cause of sudden in-hospital death. Studies have shown that the immune/inflammatory response plays an important role in the pathogenesis of vascular disease, with representative markers in the blood including the neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), monocyte/lymphocyte ratio (MLR), systemic immune/inflammatory index (SII), etc. However, there is a variety of immune/inflammatory indicators. Moreover, most previous studies have been single-center investigations involving one or two indicators, with varying nature of cases, number of cases and study objectives, thereby making it difficult to reach consensus conclusions with good clinical guidelines. This article reviews the clinical value of immunoinflammatory indicators for VTE based on previous studies, including the diagnostic and prognostic capabilities. In conclusion, NLR provides promising predictive capability for the onset and prognosis of VTE and deserves extensive application in clinical practice. PLR also has certain diagnostic and prognostic value, but further studies are warranted to identify its reliability and stability. Monocytes, eosinophils and platelet-related indicators show some clinical association with VTE, although the predictive capabilities are mediocre. SII is of promising potential value for VTE and deserves further investigations. This review will provide new clues and valuable clinical guidance for the diagnosis and therapy of VTE.Keywords: venous thromboembolism, immune/inflammation biomarker, diagnosis, prognosis

Guiying Jiang,1,* Xueqian Wang,1,* Ying Zhou,1 Chenming Zou,2,3 Ling Wang,1 Wei Wang,1 Danya Zhang,1 Hanjie Xu,1 Jie Li,1 Fei Li,1 Danfeng Luo,1 Xiangyi Ma,1 Ding Ma,1 Songwei Tan,2 Rui Wei,1 Ling Xi1 1Department of Obstetrics & Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei Province, People’s Republic of China; 2Tongji School of Pharmacy, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei Province, People’s Republic of China; 3School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ling Xi; Rui WeiDepartment of Obstetrics & Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei, People’s Republic of ChinaTel/Fax +86 27 83662688Email lxi@tjh.tjmu.edu.cn; weirui2018@tjh.tjmu.edu.cnBackground and Purpose: Cisplatin–paclitaxel (TP) combination chemotherapy as the first-line therapy for numerous cancers is hindered by its inadequate accumulation in tumors and severe side effects resulting from non-specific distribution. The aim of this study is to explore whether TMTP1-modified, cisplatin and paclitaxel prodrugs co-loaded nanodrug could improve cervical cancer chemotherapy and relieve its side effects through active and passive tumor targeting accumulation and controlled drug release.Methods: TDNP, with capacities of active targeting for tumors and controlled drug release, was prepared to co-deliver cisplatin and paclitaxel prodrugs. The characteristics were investigated, including the diameter, surface zeta potential, stability and tumor microenvironment (TME) dependent drug release profiles. Cellular uptake, cytotoxicity, drug accumulation in tumors, antitumor effects and safety analysis were evaluated in vitro and in vivo.Results: The oxidized cisplatin and the paclitaxel linked to the polymer achieved a high loading effciency of over 80% and TME-dependent sustained drug release. Moreover, TMTP1 modification enhanced cellular uptake of TDNP and further improved the cytotoxicity of TDNP in vitro. In vivo, TDNP showed an extended blood circulation and increased accumulation in SiHa xenograft models with the aid of TMTP1. More importantly, TDNP controlled tumor growth without life-threatening side effects.Conclusion: Our study provided a novel TP co-delivery platform for targeted chemotherapy of cervical cancer, which was promising to improve the therapeutic effcacy of TP and may also have application in other tumors.Keywords: TME-responsive, targeted co-delivery, combined chemotherapy, cervical cancer

Da Ma, Zhi-Guang Yao, Yan-Ping Guo, Rui-Fa Wu Department of Pediatric General Surgery, Dongguan Eighth People’s Hospital (Dongguan Children’s Hospital), Dongguan, 523000, People’s Republic of ChinaCorrespondence: Da MaDepartment of Pediatric General Surgery, Dongguan Eighth People’s Hospital (Dongguan Children’s Hospital), Dongguan, People’s Republic of ChinaTel/Fax +86 769-86186253Email mada66_doc@163.comObjective: Cryptorchidism is the most common congenital disability and is defined as the absence of one or both testicles in the scrotum during fetal development. In patients with cryptorchidism, lowered serum inhibin B levels suggest testicular dysfunction. The practical application of serum inhibin B levels in childhood remains controversial. The purpose of this study was to observe the post-operational changes of serum inhibin B levels in patients with cryptorchidism and provide evidence that serum inhibin B is a potential marker for the efficacy of testicular surgery.Methods: A total of 83 male children with cryptorchidism were enrolled in this study. All the children had blood samples taken by venipuncture on the day of surgery. The serum inhibin B levels in the blood samples were measured using an ELISA kit. Paired t-tests were used to assess differences between the groups.Results: The results show that, compared with the preoperative values, there was a statistically significant increase in serum inhibin B values one month after the operation in each group (P < 0.05). Compared with the one-month post-operation results, there were no significant changes six months after operation in each group (P> 0.05). However, serum inhibin B values decreased significantly in all groups at 12 months post-operation (P < 0.05), suggesting that serum inhibin B values are significantly correlated with age. Correlation analysis showed that serum inhibin B was negatively correlated with age from 6 to 36 months, and the correlation coefficient was − 0.488.Conclusion: Serum inhibin B can be used as a short-term evaluation index of cryptorchidism surgery rather than as a long-term indicator.Keywords: serum inhibin B, children, cryptorchidism surgery

Bin Zheng,1,* Yakun Yang,1,* Jinghan Li,1 Jing Li,1 Saijie Zuo,1 Xi Chu,2 Shan Xu,3 Donglai Ma,1 Li Chu1,4 1School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 2Department of Pharmacy, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050011, People’s Republic of China; 3Hebei Province Hospital of Chinese Medicine, Affiliated Hospital of Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 4Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China*These authors contributed equally to this workCorrespondence: Donglai Ma; Li ChuSchool of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of ChinaTel/Fax +86 311 89926719Email mdl_hebei@aliyun.com; chuli0614@126.comPurpose: Magnesium isoglycyrrhizinate (MgIG), a single stereoisomer magnesium salt of glycyrrhizic acid, has beneficial effects on the cardiovascular system through anti-inflammatory, anti-oxidation, and anti-apoptotic actions. However, MgIG has not been shown to provide protection against cardiotoxicity induced by arsenic trioxide (ATO). This study aims to demonstrate the protection of MgIG against ATO-induced cardiac toxicity in mice and to investigate the underlying mechanism.Methods: A mouse cardiotoxicity model was established by administering 5 mg/kg ATO for 7 days. MgIG used in conjunction with the ATO to assess its cardioprotection.Results: MgIG administration could significantly reduce reactive oxygen species generation and the changes in tissue morphology. Also, MgIG administration increased the activity of antioxidase, such as superoxide dismutase, catalase, and glutathione peroxidase, and reduced malondialdehyde content and pro-inflammatory cytokine levels. Western blotting showed decreased expression of Bcl-2 associated X protein and Caspase-3, with increased expression of B-cell lymphoma 2. Importantly, MgIG administration increased nuclear factor-erythroid-2-related factor 2 (Nrf2) expression, while the expressions of nuclear factor kappa-B (NF-κB) and toll-like receptor-4 (TLR4) were significantly decreased.Conclusion: Our data showed that MgIG alleviates ATO-induced cardiotoxicity, which is associated to the anti-inflammation, anti-oxidation, and anti-apoptosis action, potentially through activation of the Nrf2 pathway and suppression of the TLR4/NF-κB pathway.Keywords: magnesium isoglycyrrhizinate, arsenic trioxide, cardiotoxicity, Nrf2, TLR4/NF-κB

Qingli Cui, Yanhui Hu, Dongyang Ma, Huaimin Liu Department of Integrated Traditional and Western Medicine, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou City, Henan Province, People’s Republic of ChinaCorrespondence: Huaimin LiuDepartment of Integrated Traditional and Western Medicine, The Affiliated Cancer Hospital of Zhengzhou University, 127 Dongming Road, Zhengzhou 450008, People’s Republic of ChinaEmail huaiminliu@sina.comObjective: Anlotinib, an oral small-molecular tyrosine kinase inhibitor (TKI) on tumor angiogenesis and growth, has a wide spectrum of inhibitory effects on targets such as vascular endothelial growth factor receptors 2/3 (VEGFR2/3), etc. The efficacy and safety of anlotinib in the treatment of platinum-resistant or platinum-refractory ovarian cancer were evaluated.Patients and Methods: Patients with platinum-resistant or platinum-refractory ovarian cancer that treated with anlotinib in the Affiliated Cancer Hospital of Zhengzhou University from May 2018 to March 2020 were included. Medical records were reviewed in terms of objective response, survival outcomes, and safety.Results: A total of 38 patients were analyzed. The median progression-free survival and the median overall survival were 7.7 months (95% CI: 6.7– 8.7) and 16.5 months (95% CI: 13.3– 19.7), respectively. About 17 patients received anlotinib monotherapy, and the median progression-free survival was 7.7 months (95% CI: 6.3– 9.1). A total of 19 cases received anlotinib plus chemotherapy with a median progression-free survival of 8.0 months (95% CI: 4.8– 11.2). A total of 2 cases received anlotinib plus anti-PD-1 antibody pembrolizumab, and 1 case had partial response, the other progressive disease. The objective response rate was 42.1% while the disease control rate was 86.8%. A total of 5 patients experienced dose reduction from 12 mg to 10 mg because of adverse effects. The most common adverse effects were hypertension (31.6%), fatigue (28.9%), anorexia (26.3%) and hand-foot syndrome (23.7%). No treatment-related deaths were recorded.Conclusion: Anlotinib produced moderate improvements in progression-free survival and overall survival in patients with platinum-resistant or platinum-refractory ovarian cancer. It indicates that anlotinib maybe a new treatment option for patients with platinum-resistant or platinum-refractory ovarian cancer.Keywords: angiogenesis, anlotinib, ovarian cancer, platinum-resistant

Luoning Gou,1,* Ming Xiang,2,* Xiao Ran,3,* Fen Wang,1 Shujun Zhang,1 Shusheng Li,3 Kun Dong,1 Xi Chen,1 Yangxin Huang,4 Chengzhen Meng,2 Qian Fan,2 Yan Yang,1 Xuefeng Yu,1 Delin Ma,1 Ping Yin2 1Department of Endocrinology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 2Department of Epidemiology and Biostatistics, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 3Department of Emergency Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 4College of Public Health, University of South Florida, Tampa, Florida, The United States*These authors contributed equally to this workCorrespondence: Ping Yin; Delin MaTongji Medical College, Huazhong University of Science and Technology, 13 Hongkong Road, Wuhan, 430030 Hubei, People’s Republic of ChinaTel + 86-27-83692832Fax +86-27-83692333Email pingyin2000@126.comDelin Ma Email maderine4@163.comPurpose: Recently, a cluster of pneumonia caused by SARS-CoV-2 were identified in Wuhan and spread throughout the world. More information about risk factors for mortality of critically ill patients infected with SARS-CoV-2 remain to be evaluated.Methods: We included adult patients confirmed with SARS-CoV-2 infection who were critically ill and admitted to the intensive care unit (ICU) of Tongji Hospital in Wuhan from Feb 4, 2020 to Feb 20, 2020. Data were collected and compared between patients who died and improved. Logistic regression was used to explore the risk factors for death of SARS-CoV-2-infected critically ill patients.Results: A total of 160 critically ill patients with SARS-CoV-2 infection were included, of which 146 patients with appeared outcomes were included into the final analysis. The random blood glucose, serum sodium and effective plasma osmolarity were higher in deceased patients, especially in patients with diabetes. There were 7 patients with diabetes with hyperosmolar status and all of them were deceased. Multivariable regression revealed that older age (odds ratio 4.28, 95% CI 1.01– 18.20; p = 0.049), higher C-reactive protein (odds ratio 1.01, 1.00– 1.03; p = 0.024), higher interleukin-6 (odds ratio 1.01, 1.00– 1.03; p = 0.0323), and d-dimer greater than 1 μg/mL (odds ratio 1.10, 1.01– 1.20; p = 0.032) at admission were associated with increased odds of death.Conclusion: In conclusion, hyperosmolarity needs more attention and may contribute to mortality in critically ill patients with COVID-19, especially in those with diabetes. Older age, inflammatory response, and thrombosis may be risk factors for death of critically ill patients with SARS-CoV-2 infection.Keywords: SARS-CoV-2, critically ill, hyperosmolarity, diabetes, risk factors, mortality

Xin Zheng,1,2,* Yun Chen,3,* Danyan Ma,4 Mulin Zhang,1,2 Yinxiang Huang,1,2 Meifeng Tong,1,2 Bing Yan,1,2 Shaowei Lin,5 Xiaohong Yan,6 Changqin Liu1– 3 1Department of Endocrinology and Diabetes, The First Affiliated Hospital of Xiamen University, Xiamen, China; 2Fujian Province Key Laboratory of Diabetes Translational Medicine, Xiamen Clinical Medical Center for Endocrine and Metabolic Diseases, Xiamen, China; 3The School of Clinical Medicine, Fujian Medical University, Fuzhou, China; 4School of Medicine, Xiamen University, Xiamen, China; 5Department of Nuclear Medicine, The First Affiliated Hospital of Xiamen University, Xiamen, China; 6Reproductive Medicine Center, The First Affiliated Hospital of Xiamen University, Xiamen, China*These authors contributed equally to this workCorrespondence: Changqin LiuDepartment of Endocrinology and Diabetes, The First Affiliated Hospital of Xiamen University, Xiamen, ChinaTel +86-592-2137796Fax +86-592-2137189Email liuchangqin@xmu.edu.cnXiaohong YanReproductive Medicine Center, The First Affiliated Hospital of Xiamen University, Xiamen, ChinaTel +86-592-2662318Fax +86-592-2662318Email xmyxh1224@sina.comObjective: The aim of the present study was to investigate the possible correlation between the percentage of daily energy intake from fat (PEF) with insulin resistance (IR) in women with polycystic ovary syndrome (PCOS).Methods: In this cross-sectional study, a total of 186 females with PCOS were screened. Daily dietary intake data were collected by a trained nutritionist using the 24-h dietary recall method over three consecutive days. A total of 111 subjects who had complete data were divided into two groups based on the percentage of daily energy intake from fat (PEF): the normal PEF (NPEF) group (PEF < 30%) and the high PEF (HPEF) group (PEF ≥ 30%). Pearson’s correlation analysis and stepwise multivariate linear regression analysis were used to analyze the correlation of PEF with homeostasis model assessment of insulin resistance (HOMA-IR).Results: The total prevalence rate of overweight/obesity was 80.2%. There were significant differences in waist circumference (WC), body mass index (BMI), fasting insulin, and HOMA-IR (P < 0.001) among the normal weight, the overweight, and the obese groups, but no significant differences were observed in total energy and dietary macronutrients intake in the three groups. The daily intake of fat and protein, fasting insulin, and HOMA-IR in the NPEF group were significantly higher than those in the HPEF group. Pearson’s correlation analysis showed PEF in PCOS women was negatively correlated with BMI (r= − 0.189, p=0.047) and HOMA-IR (log-transformed) (r= − 0.217, p=0.022). Further, stepwise multivariate linear regression analysis showed PEF was negatively correlated with HOMA-IR (p< 0.05).Conclusion: The percentage of daily energy intake from fat is negatively correlated with IR in women with PCOS.Keywords: the percentage of daily energy intake from fat, polycystic ovary syndrome, insulin resistance, daily dietary intake, obesity

Danxu Ma,1 Huili Li,1 Rong Shi,1 Yinan Yang,1 Haoda Liu,2 Xiaodong Ge1 1Department of Anesthesiology, Beijing Chaoyang Hospital, Capital Medical University, Beijing, People’s Republic of China; 2Department of Anesthesiology, Beijing Huairou Maternal and Child Health Care Hospital, Beijing, People’s Republic of ChinaCorrespondence: Xiaodong GeDepartment of Anesthesiology, Beijing Chaoyang Hospital, Capital Medical University, No. 8, Gongtinan Road, Chaoyang District, Beijing 100020, People’s Republic of ChinaEmail xiaodongge2005@163.comBackground: Persistent or recurring radicular pain after lumbar surgery is a clinical condition of failed back surgery syndrome (FBSS) that seriously affects the life quality of patients. Conventional medication and physiotherapy do not fully relieve this pain. A simpler, safer, and less invasive option is lumbar selective nerve root block or paravertebral block. Here, we share our experience regarding lumbar paravertebral block for a patient with FBSS, which successfully alleviated radicular pain after lumbar surgery.Case Presentation: An 80-year-old man with left lower limb radicular pain diagnosed as L4-5, L5-S1 intervertebral disc protrusion, spinal canal stenosis, and degenerative scoliosis underwent lumbar surgery. Four months after surgery, he experienced left lower limb radicular pain. After designing the puncture route based on X-ray film, we performed a combined ultrasound-guided L4 and L5 paravertebral block. With his improved pain control, his functional status and ability to perform daily activities also markedly improved.Conclusion: Real-time ultrasound-guided lumbar paravertebral block performed with a pre-designed route on X-ray film can provide a simple and safe way to relieve radicular pain in FBSS.Keywords: lumbar paravertebral block, failed back surgery syndrome, ultrasound-guided, pre-designed route

Hong-Peng Chen,1,2,* Xiao-Yan Wang,1,* Xiao-Yan Pan,1 Wang-Wang Hu,1 Shu-Ting Cai,1 Kiran Joshi,3 Lie-Hua Deng,1,2 Daqing Ma3 1Department of Critical Care Medicine, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, People’s Republic of China; 2Zhanjiang Key Laboratory of Organ Injury and Protection and Translational Medicine, Zhanjiang, Guangdong, People’s Republic of China; 3Division of Anesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea and Westminster Hospital, London, UK*These authors contributed equally to this workCorrespondence: Lie-Hua Deng; Daqing MaDepartment of Critical Care Medicine, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, ChinaEmail glinson@126.com; d.ma@imperial.ac.ukIntroduction: Because of its high morbidity and mortality, sepsis remains the leading cause of death in the ICU. Microparticles (MP) have been largely studied as potential diagnostic or prognostic markers in various diseases including sepsis.Objective: The biological and clinical relevance of neutrophil-derived microparticles (NDMPs) within the MP population remains unclear. The objective of this study was to elucidate the relationship between plasma NDMPs and the prognosis of patients with sepsis and/or septic shock.Methods: The study was designed as an observational, noninterventional clinical study. The cohort for this study included 40 sepsis and 40 septic shock patients together with 10 healthy controls admitted to the Intensive Care Unit (ICU) and the Health Surveillance Center in the Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, China, from January to November 2018, respectively. The degree of critical disease for sepsis and septic shock was evaluated, with data analyses conducted from 2018 to 2019.Results: On days 1, 3 and 5 post-admission a series of data including plasma NDMP levels, patient demographics, TNF-α levels, IL-6 levels, sTREM-1 levels, and the sepsis severity score measurements were collected. A survival curve was plotted against levels of plasma NDMPs. Levels of NDMPs were observed to be higher in the septic shock patients than in the sepsis patients on days 1, 3, and 5 post-ICU admission (p < 0.05). NDMP levels were significantly increased in sepsis and septic shock patients with a parallel increase in pro-inflammatory mediators and sepsis severity score (p < 0.05) as well as mortality.Conclusion: Our data suggest that NDMPs may be a biomarker of sepsis severity and mortality although its implications on sepsis prognosis warrant further study.Keywords: sepsis, septic shock, cell-derived microparticles, biomarker, prognosis

Rong Shi, Danxu Ma, Yinan Yang, Yun Wang Department of Anesthesia and Pain Medicine, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, People’s Republic of ChinaCorrespondence: Yun WangDepartment of Anesthesia and Pain Medicine, Beijing Chaoyang Hospital, Capital Medical University, No. 8, Gongtinan Road, Chaoyang District, Beijing 100020, People’s Republic of ChinaEmail wangyun129@ccmu.edu.cnBackground: Imaging-guided celiac plexus neurolysis using ultrasound (US) guidance via a transabdominal approach and endoscopic-ultrasound (EUS) has been increasingly applied for the treatment of pancreatic cancer-associated abdominal pain.Objective: To investigate the application of ultrasound-guided and fluoroscopy-assisted celiac plexus neurolysis in a patient with advanced pancreatic cancer suffering from refractory abdominal pain for which oral opioid treatment was ineffective.Case Report: We report a case of ultrasound-guided and fluoroscopy-assisted celiac plexus neurolysis in a patient with advanced pancreatic cancer with refractory abdominal pain. With the patient in the prone position, celiac plexus neurolysis was performed under real-time US guidance. The transducer was placed below the costal margin and a puncture needle with an ultrasound enhancement tip was inserted in-plane aiming for the lateral anterior end of the vertebral body. The correct needle tip position was confirmed by the C-arm with contrast material located anterior to the vertebral body and posterior to the diaphragm.Conclusion: We highlight the use of an US-guided and fluoroscopy-assisted posterior approach for use in celiac plexus neurolysis procedures, particularly in patients suffering from contraindications from the US or EUS-guided anterior approaches.Keywords: ultrasound, image-guidance, posterior approach, celiac plexus neurolysis, pancreatic cancer, abdominal pain

Yongheng Ye1 ,* Lingli Zhang2 ,* Yuhu Dai,1 Zhi Wang,3 Cuie Li,3 Yue Peng,4 Dong Ma,2 Peiheng He1 1Department of Orthopedic Surgery, Guangdong Provincial Key Laboratory of Orthopaedics and Traumatology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangzhou, Guangdong Province 510080, People’s Republic of China; 2Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou 510632, People’s Republic of China; 3Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong Province 510530, People’s Republic of China; 4Department of Otorhinolaryngology Head and Neck Surgery, Zhuhai People’s Hospital (Zhuhai Hospital Affiliated with Jinan University), Zhuhai, Guangdong Province 519000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Peiheng HeDepartment of Orthopedic Surgery, Guangdong Prov Key Laboratory Orthopaed & Traumatol, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong Province 510080, People’s Republic of ChinaEmail hepeiheng@mail.sysu.edu.cnDong MaKey Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou 510632, People’s Republic of ChinaEmail tmadong@jnu.edu.cnObjective: This study aimed to develop aptamer-anchored hyperbranched poly(amido amine) (HPAA) for the systemic delivery of miRNA-133a-3p and to evaluate its therapeutic potential against bone metastasis of prostate cancer in vivo and in vitro.Methods: A glutathione (GSH)-responsive cationic HPAA was prepared by the Michael addition reaction. Furthermore, HPAA-PEG was produced by PEGylation, and then the aptamer targeted to prostate-specific membrane antigen (PSMA) was conjugated to the HPAA-PEG. The obtained HPAA-PEG-APT could form nanocomplexes with miRNA-133a-3p through electrostatic adsorption.Results: The results of immunocytochemistry indicated that the complexes could target PSMA-expressing LNCaP cells. The ability of HPAA-PEG-APT to facilitate the delivery of miRNA-133a-3p into LNCaP cells was proven, and HPAA-PEG-APT/miRNA-133a-3p demonstrated enhanced antitumor activity, lower cytotoxicity and better biocompatibility in vitro. Moreover, in a mouse tibial injection tumor model, the intravenous injection of the HPAA-PEG-APT/miRNA-133a-3p complex significantly inhibited cancer growth and extended the survival time.Conclusion: This study provided an aptamer-anchored HPAA-loaded gene system to deliver miRNA-133a-3p for better therapeutic efficacy of bone metastasis of prostate cancer.Keywords: miRNA, aptamer, hyperbranched polyamide amine, bone metastasis, prostate cancer

Donghe Ma,1 Yan Zhang,2 Guanghua Chen,3 Jia Yan1 1Digestive Department, University-Town Hospital of Chongqing Medical University, Chongqing 401331, People’s Republic of China; 2Neurology Department, People’s Hospital of Hechuan Chongqing, Chongqing 401520, People’s Republic of China; 3Dermatology Department, Children’s Hospital of Chongqing Medical University, Chongqing 400014, People’s Republic of ChinaCorrespondence: Jia Yan Tel +86-17782351875Fax +86-23-65715900Email yanjia19850730@163.comPurpose: The objective of this study was to investigate the effect of miR-148a on the polarization and recruitment of tumor-associated macrophages (TAMs).Methods: In human monocyte THP-1 cells, M1 or M2 differentiation was induced by phorbol 12-myristate 13-acetate (PMA) with specific induction supplements and identified using flow cytometry and ELISA. To alter cellular miR-148a expression level, THP-1 cells were transfected with miR-148a mimics or inhibitors. A dual-luciferase assay was used to determine whether miR-148a could directly regulate the expression of signal regulatory protein α (SIRPα). Expression of miR-148a and SIRPα was detected with RT-PCR or Western blot. A co-culture system of THP-1 cells and colorectal cancer SW480 cells was used for TAM induction. The recruitment of macrophage to SW480 cells was measured using chemotaxis assay. In SW480 cells, apoptosis induced by macrophages was detected using flow cytometry and a xenograft assay. Macrophage infiltration was detected by immunofluorescence assay in tumor tissues.Results: miR-148a over-expression increased M1-related CD86 expression in THP-1 cells and promoted differentiation to M1-like macrophages. Inhibition of miR-148a increased M2-related CD206 expression and promoted differentiation to M2-like macrophages. In the co-culture system, THP-1 cells were induced to the M2-like state by SW480 cells. The level of miR-148a negatively correlated with the levels of M2-related cytokines. Additionally, miR-148a expression level was negatively associated with macrophage recruitment by colorectal cancer cells. Furthermore, in miR-148a over-expression, the number of macrophages recruited by SW480 cells was reduced. Meanwhile, cancer cell apoptosis induced by macrophages was enhanced. Thus, miR-148a expression was beneficial for the transformation of macrophages, from an immune-suppressive status to an immune-promoting status. These anti-cancer effects of miR-148a were related to the down-regulation of SIRPα in macrophages, directly targeted by miR-148a. A xenograft assay showed that the co-inoculation of macrophages over-expressing miR-148a reduced subcutaneous tumorigenesis and M2 macrophage infiltration.Conclusion: miR-148a promoted the differentiation of M0 macrophages into anti-tumor classical activation type M1, and reduced TAM recruitment by targeting SIRPα to inhibit colorectal cancer cell viability.Keywords: colorectal cancer, tumor-associated macrophage, microRNA-148a, polarization, signal regulatory protein alpha

Qi Jia,1 Fan Jiang,2 Daliang Ma,1 Jun Li,1 Fan Wang,1 Zhiqiang Wang1 1Encephalopathy Center, The Third People’s Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830000, People’s Republic of China; 2Neonatal Intensive Care Unit, The First People’s Hospital of Urumqi, Urumqi 830000, People’s Republic of ChinaCorrespondence: Zhiqiang WangEncephalopathy Center, The Third People’s Hospital of Xinjiang Uygur Autonomous Region, No. 168, Nanchang Road, Urumqi 830000, People’s Republic of ChinaTel/ Fax +86-9917530284Email xjwjwzq@163.comPurpose: To study the association between IL-6 level and seizure recurrence in patients with the first post-ischemic stroke seizure and assess its predictive value for seizure recurrence.Patients and Methods: A total of 2976 consecutive ischemic stroke patients were retrospectively enrolled. Among them, 209 (7.02%) patients with the first post-ischemic stroke seizure were included in this analysis. The IL-6 mRNA expression level was evaluated through quantitative real-time PCR (qRT-PCR) and the 2−ΔΔCt method. Demographic data and clinical characteristics were collected. Univariate analysis was performed with independent-samples t-test, Mann–Whitney U-test, or chi-square test. Multivariate analysis was conducted using a backward stepwise logistic regression model for variables with P< 0.10 in univariate analysis. The predictive value was assessed using a receiver operating characteristic (ROC) curve.Results: Among the patients included, 105 (50.24%) had recurrence of seizures, and 104 (49.76%) had no recurrence. Multivariate analysis demonstrated that the IL-6 mRNA expression level was independently correlated with seizure recurrence in patients with the first post-ischemic stroke seizure after adjusting for age, NIHSS scores, time of seizure, seizure type, lesion size, location of the offending lesion to different hemispheric lobes, cortical involvement, gender, electroencephalography (EEG) findings, and hemorrhagic transformation. When the IL-6 mRNA expression level was used to predict seizure recurrence, the area under the ROC curve (AUC) was 0.763 (SE=0.033, 95% CI=0.698– 0.829). The diagnostic power was moderate.Conclusion: IL-6 was independently correlated with seizure recurrence in patients with the first post-ischemic stroke seizure and might be a potential biomarker for prediction of seizure recurrence.Keywords: post-ischemic stroke seizure, seizure recurrence, IL-6, predictive value

Soyeon Bang,1 Junhong Li,2 Meiqin Zhang,3 Renjie Cui,3 Xingwen Wu,4 Zhaochen Xin,1 Duan Ma,3,5 Jin Zhang,3 Hongwei Zhang1 1Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; 2Department of Urology, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, People’s Republic of China; 3Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, Shanghai, People’s Republic of China; 4Department of Dentistry, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; 5Children’s Hospital, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Hongwei ZhangDepartment of General Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, People’s Republic of ChinaTel +86-13681973106Fax +86 21-64038472Email zhang.hongwei@zs-hospital.sh.cnDuan MaKey Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, 130 Dong an Road, Xuhui District, Shanghai 200032, People’s Republic of ChinaTel +86-13916126452Fax +86 21-54237441Email duanma@shmu.edu.cnBackground: Krüppel-like factor 16 (KLF16), a member of the KLF family, is involved in metabolism and regulation of the endocrine system and has emerging roles in tumor progression. However, the expression of KLF16 and its role in breast cancer are elusive.Methods: We investigated the expression and prognostic value of KLFs in breast cancer using data acquired from the TCGA BRCA dataset and the Kaplan–Meier plotter dataset. The protein levels of KLF16 in breast specimens were detected by immunohistochemistry (IHC). KLF16 silencing using shRNAs was performed to explore the effects of KLF16 on breast cancer cell growth, migration, and invasion. The expression of EMT markers in cells manipulated for KLF16 expression was assessed by Western blotting.Results: Using publicly available dataset and specimens from breast cancer patients, we found that the expression levels of KLF16 were significantly higher in tumor tissues and that high levels of KLF16 were associated with poor prognosis in breast cancer patients. Moreover, KLF16 expression levels had relation to several clinicopathological parameters of breast cancer, including the molecular subtype and histological grade. Importantly, knockdown of KLF16 dramatically suppressed cell proliferation both in vitro and in vivo. Also, KLF16 deletion impaired migration, and invasion in breast cancer cells, and suppressed epithelial–mesenchymal transition (EMT).Conclusion: Our results suggest that KLF16 has important oncogenic functions in breast cancer and that the expression levels of KLF16 are associated with prognosis in breast cancer patients. Our findings also suggest that KLF16 is involved in proliferation, migration, and invasion in breast cancer cells. Thus, KLF16 might be a promising prognostic marker and a therapeutic target for breast cancer.Keywords: breast cancer, EMT, Krüppel-like factor 16, metastasis, proliferation

Lin Zhu,1 Bo Yang,2 Dongxia Ma,3 Lan Wang,4 Wu Duan5 1Department of Pediatrics, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology, Wuhan 430030, People’s Republic of China; 2Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People’s Republic of China; 3Department of Allergy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 4Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology, Wuhan 430030, People’s Republic of China; 5Division of Endocrinology, Department of Internal Medicine, Qilu Hospital of Shandong University, Jinan 250012, People’s Republic of ChinaCorrespondence: Wu DuanDivision of Endocrinology, Department of Internal Medicine, Qilu Hospital of Shandong University, Jinan 250012, People’s Republic of ChinaTel/ Fax +86-531-8692-7544Email duan5wu@qq.comAbstract: Hydrogen sulfide (H2S) is now increasingly considered to be the third gasotransmitter alongside other gaseous signaling molecules, nitric oxide (NO) and carbon monoxide (CO). H2S is produced by a variety of endogenous enzymatic and non-enzymatic pathways and acts as a modulator of the physiological and pathological events of the body. Adipocytes express the cystathionine γ lyase (CSE)/H2S system, which modulates a variety of biological activities in adipose tissue (AT), including inflammation, apoptosis, insulin resistance, adipokine secretion and adipocyte differentiation. Abnormalities in the physiological functions of AT play an important role in the process of diabetes mellitus. Therefore, this review provides an overview of the general aspects of H2S biochemistry, the effect of H2S on AT function and diabetes mellitus and its molecular signalling mechanisms as well as the potential application of H2S in pharmacotherapy.Keywords: hydrogen sulfide, adipose tissue, diabetes mellitus, drug therapy

Jun Jiao,1 Xinlin Jiao,1 Qingqing Liu,1 Wenjie Qu,1 Daoxin Ma,2 Youzhong Zhang,1 Baoxia Cui1 1Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan 250012, People’s Republic of China; 2Hematology Oncology Center, Qilu Hospital, Shandong University, Jinan 250012, People’s Republic of ChinaCorrespondence: Baoxia Cui Tel +8618560081862Email cuibaoxia@sdu.edu.cnPurpose: Accumulating evidence indicates that circular RNAs (circRNAs) are closely involved in canceration and cancer progression. However, the role of circRNAs in cervical cancer (CC) is largely unknown. Here, we characterized the role of circRNA_101308 in CC.Materials and Methods: The expression of circRNA_101308 in CC tissues was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Then, circRNA_101308 was overexpressed in CC cells to detect its function by proliferation and apoptosis assays, Transwell assays and animal experiments. The potential mechanism of circRNA_101308 in CC was explored by RNA pull-down, Gene Ontology (GO) and pathway analyses.Results: CircRNA_101308 was significantly downregulated in CC tissues. The level of circRNA_101308 was much lower in CC patients with lymph node metastasis or deep myometrial invasion compared to those patients without lymph node metastasis and superficial myometrial invasion. CircRNA_101308 overexpression inhibited CC cell proliferation, invasion and migration. MiR-26a-5p, miR-196a-5p, miR-196b-5p, miR-335-3p, and miR-1307-3p were found to be sponged by circRNA_101308 in CC cells. Further, GO and pathway analyses predicted the potential functional processes and pathways of circRNA_101308 in CC.Conclusion: CircRNA_101308 is downregulated and acts as a tumor suppressor in CC. CircRNA_101308 can participate in many different processes by sponging different miRNAs in CC cells. This exploration of circRNA_101308 provides new directions for research on cancer development and the clinical treatment of CC.Keywords: circular RNA, cervical cancer, circRNA_101308, microRNA

Yanshuang Liu,1,2,* Yingran Liang,3,* Bin Zheng,3 Li Chu,3 Donglai Ma,3 Hongfang Wang,3 Xi Chu,4 Jianping Zhang2,5 1Department of Diagnostics, School of Integrated Chinese and Western Medicine, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 2Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns, Shijiazhuang 050200, Hebei, People’s Republic of China; 3Department of Pharmaceutics, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China; 4Department of Pharmacy, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050011, People’s Republic of China; 5Department of Pharmacology, School of Basic Medicine, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xi ChuThe Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, People’s Republic of ChinaTel/Fax +86 311 86095324Email chux2014@126.comJianping ZhangSchool of Basic Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, People’s Republic of ChinaTel/Fax +86 311 89926719Email zhangjianping14@126.comPurpose: Arsenic trioxide (ATO) has been shown to induce hepatic injury. Crocetin is a primary constituent of saffron, which has been verified to have antioxidant and anti-inflammatory effects. In the current experiment, we evaluated the efficacy of crocetin against ATO-induced hepatic injury and explored the potential molecular mechanisms in rats.Methods: Rats were pretreated with 25 or 50 mg/kg crocetin 6 h prior to treating with 5 mg/kg ATO to induce hepatic injury daily for 7 days.Results: Treatment with crocetin attenuated ATO-induced body weight loss, decreases in food and water consumption, and improved ATO-induced hepatic pathological damage. Crocetin significantly inhibited ATO-induced alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) increases. Crocetin prevented ATO-induced liver malondialdehyde (MDA) and reactive oxygen species (ROS) levels. Crocetin abrogated the ATO-induced decrease of catalase (CAT) and superoxide dismutase (SOD) activity. Crocetin was found to significantly restore the protein levels of interleukin 6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor-alpha (TNF-α). Furthermore, crocetin promoted the expression of nuclear factor erythroid 2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADP(H): quinone oxidoreductase 1 (NQO1).Conclusion: These findings suggest that crocetin ameliorates ATO-induced hepatic injury in rats. In addition, the effect of crocetin might be related to its role in antioxidant stress, as an anti-inflammatory agent, and in regulating the Nrf2 signaling pathway.Keywords: crocetin, arsenic trioxide, hepatotoxicity, oxidative stress, inflammation, Nrf2 signaling pathway

Luo Zhao, Yingzhi Qin, Dongjie Ma, Li Li, Zhijun Han, Shanqing Li, Hongsheng Liu Department of Thoracic Surgery, Peking Union Medical College Hospital, CAMS & PUMC, Beijing 100730, People’s Republic of ChinaCorrespondence: Hongsheng LiuDepartment of Thoracic Surgery, Peking Union Medical College Hospital, Beijing 100730, People’s Republic of ChinaTel +86 18519667758Fax +86 10 69152630Email pumchlhsnj@163.comPurpose: To assess the use of video-assisted thoracoscopic surgery to treat lung metastases from refractory choriocarcinoma.Patients and Methods: We reviewed patients diagnosed with refractory choriocarcinoma who underwent lung resection by video-assisted thoracoscopic surgery combined with chemotherapy between October 2013 and August 2019 at the Peking Union Medical College Hospital. The surgical records, pathologic findings and survival rates were analyzed.Results: The study included 73 patients who underwent 78 thoracoscopic surgeries. Most patients underwent lobectomy (48.7%), and 17 patients (21.8%) underwent resection of more than one lobe. The median operation time and bleeding volume were 95 minutes and 50 mL, respectively. The median duration of chest tube use and hospital stay were 3 days and 4 days, respectively. Postoperative complications were documented in 6 patients (7.7%). The thoracic lymph nodes were harvested in 51 patients (65.4%), but none of these patients had positive nodes. A total of 69.2% of the patients had positive pathologic findings. The mean follow-up time was 30 months. During follow-up, 11 patients experienced disease relapse, and 2 of them died because of brain metastasis. The overall disease-free rate was 83.6%, and the survival rate was 97.0% after excluding those lost to follow-up. Patients with decreased postoperative β-hCG showed a higher disease-free rate during follow up (P< 0.05).Conclusion: The minimally invasive video-assisted thoracoscopic approach is a valuable and safe treatment for refractory choriocarcinoma patients with lung metastases. Lymphadenectomy is not suggested for these patients. Patients with decreased postoperative β-hCG levels may achieve a much better prognostic result.Keywords: video-assisted thoracoscopic surgery, lung metastases, choriocarcinoma

Danyan Ma,1,* Xuanlin Zheng,2,* Lianqin Dong,3 Caiyu Zheng,4 Yun Chen,3 Zheng Chen,2 MingZhu Lin,2 Xuejun Li,5 Zhibin Li,6 Changqin Liu3,7 1School of Medicine, Xiamen University, Xiamen City, Fujian Province, People’s Republic of China; 2Department of Endocrinology and Diabetes, Xiamen Clinical Medical Center for Endocrine and Metabolic Diseases, Fujian Province Key Laboratory of Diabetes Translational Medicine, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of China; 3The School of Clinical Medicine, Fujian Medical University, Fuzhou, Fujian Province, People’s Republic of China; 4Fujian Medical University Xiamen Humanity Hospital, Xiamen City, Fujian Province, People’s Republic of China; 5Department of Endocrinology and Diabetes, Xiamen Clinical Medical Center for Endocrine and Metabolic Diseases, Fujian Province Key Laboratory of Diabetes Translational Medicine, Xiamen Diabetes Institute, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of China; 6Epidemiology Research Unit, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of China; 7Department of Endocrinology and Diabetes, Xiamen Clinical Medical Center for Endocrine and Metabolic Diseases, Fujian Province Key Laboratory of Diabetes Translational Medicine, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Changqin LiuDepartment of Endocrinology and Diabetes, Xiamen Clinical Medical Center for Endocrine and Metabolic Diseases, Fujian Province Key Laboratory of Diabetes Translational Medicine, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of ChinaTel +86-592-2137610Fax +86-592-2137557Email liuchangqin@xmu.edu.cnZhibin LiEpidemiology Research Unit, The First Affiliated Hospital of Xiamen University, Xiamen City, Fujian Province, People’s Republic of ChinaTel +86-592-2137364Fax +86-592-2137557Email zhibinli33@hotmail.comObjective: To explore the association of serum vitamin-D levels with the severity of obstructive sleep apnea (OSA) in patients with type 2 diabetes mellitus (T2DM).Methods: A cross-sectional study of 136 patients with T2DM who underwent overnight polysomnography (PSG) tests and serum 25‐hydroxyvitamin‐D3 (25(OH)D3) level detections was conducted. Multivariable linear regression and logistic regression analyses were performed to determine the associations of serum 25(OH)D3 levels with apnea–hypopnea index (AHI) and obstructive sleep apnea (OSA).Results: The prevalence rates of OSA were 84.4% for male and 65.2% for female patients, respectively (p = 0.011). With increasing severities of OSA categories (none, mild, moderate and severe), patients were more likely to be male and obese, but there was no significant difference in serum 25(OH)D3 level ((mean ± standard deviation) 21.8 ± 8.8, 27.7 ± 14.6, 24.2 ± 9.8 and 26.8 ± 6.2 ng/mL, respectively, p=0.086). Serum 25(OH)D3 level was not significantly correlated with AHI (log-transformed), with the correlation coefficient of 0.133 (p=0.124). With adjustment for potential confounding factors, multivariable linear regression and logistic regression analyses showed that serum 25(OH)D3 level was not significantly associated with either AHI (log-transformed) or the risk of OSA, with the standardized regression coefficient (95% confidence interval (CI)) of 0.098 (− 0.004– 0.014, p=0.252) and the adjusted odds ratio (95% CI) of 1.055 (0.991– 1.124, p=0.095), respectively.Conclusion: Serum 25(OH)D3 level was not significantly associated with either AHI or the risk of OSA in patients with T2DM.Keywords: obstructive sleep apnea, polysomnography, 25-hydroxyvitamin-D3, apnea–hypopnea index, type 2 diabetes mellitus

Song Yao,1,* Junwei Yan,2,* Ling Li,1 Dongchun Ma,1 Jie Liu,1 Qing Wang,1 Aimin Wang,1 Fangjin Bao,1 Zekun Zhang,1 Xundi Bao1 1Tuberculosis Prevention and Treatment Institute of Anhui Province (Anhui Chest Hospital), Hefei 230022, Anhui, People’s Republic of China; 2Affiliated Psychological Hospital of Anhui Medical University, Hefei Fourth People’s Hospital, Anhui Metal Health Center, Hefei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Junwei Yan No. 316, Huangshan Road, Hefei 230022, Anhui, People’s Republic of ChinaTel/Fax +86-551-63658250Email 594347807@qq.comBackground: Multidrug-resistant tuberculosis (MDR-TB) is currently a major problem in China. The prevention and treatment work for MDR-TB patients started late in Anhui province. To determine the prevalence of MDR-TB in sputum smear-positive TB patients (SSPTBPs) and analyze the risk factors for MDR-TB in Anhui province, we conducted an investigation of drug resistance among SSPTB outpatients from September 2015 to August 2016.Methods: A stratified cluster-randomized sampling method was used to obtain a representative sample. It was estimated that 2290 new cases and 440 previously treated cases of SSPTBPs needed to be recruited from 40 survey sites. Isolates were tested for resistance to six first-line and second-line anti-TB drugs. Information from patient questionnaire survey was used to identify factors linked to MDR-TB.Results: Finally, a total of 3047 SSPTBPs were recruited from 40 survey sites; of these, 2530 specimens were successfully cultured and had drug susceptibility testing done. The proportions of rifampin resistant (RR)-TB were 11.42% (289/2530, 95% CI: 10.18– 12.66%), 7.64% (163/2133, 95% CI: 6.38– 8.62%) and 31.74% (126/397, 95% CI: 27.38– 36.60%) in all cases, new cases and previously treated cases, respectively, and the proportions of confirmed MDR-TB were 7.63% (193/2530, 95% CI: 6.59– 8.66%), 4.97% (106/2133, 95% CI: 4.05– 5.89%) and 21.91% (87/397, 95% CI: 17.83– 26.00%), respectively. The ofloxacin resistance rate in previously treated SSPTBPs reached 21.66% (95% CI: 17.33– 26.75%). Patients who had received two or more anti-TB treatment courses were significantly associated with MDR-TB compared to patients who have received one anti-TB course.Conclusion: MDR-TB prevalence was high among SSPTBPs in Anhui province, and past anti-TB treatment course was associated with MDR-TB.Keywords: drug resistance, multidrug-resistant, Mycobacterium tuberculosis, tuberculosis

Jian Cao,1,2,* Zhixu Liu,1,* Dongyang Ma,2 Shunyao Shen,1 Xudong Wang1 1Department of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai JiaoTong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, The 940th Hospital of People’s Liberation Army, Lanzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xudong WangDepartment of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai JiaoTong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, 639 Zhizaoju Road, Shanghai 200011, People’s Republic of ChinaTel +86 21 53315159Email xudongwang70@hotmail.comPurpose: Although negative pressure wound therapy (NPWT) has been widely used in complicated wound care, there are still some obstacles regarding its use in the treatment of severe deep fascial space infections in the head and neck. The purpose of this study is to describe a new modified usage of NPWT and investigate the clinical efficacy of this system in a consecutive case series of severe deep fascial space infections.Methods: The investigators implemented a new modification of NPWT for the management of severe deep fascial space infections. In this new system, the half-plugged bar-shaped foam material was arranged along with the through-and-through side-holed latex drainage tube, which could maintain negative pressure in the distal part of the spaces, and the tube was easy to remove 5– 7 days after surgery. Twelve patients had severe deep fascial space infections in the head and neck with a direct threat to the airway. The median time of removal of the NPWT device, the median amount of drainage fluid and the median healing time were investigated.Results: A total of 7 male and 5 female patients with an average age of 63.2 years old were included in this study. The median time of removal of the NPWT device was 6 days (ranging from 4 to 7 days). The median amount of drainage fluid within 3 days after surgery was 420 mL (ranging from 280– 760 mL), and the median time for complete wound healing was 12 days (ranging from 10 to 21 days).Conclusion: The results of this study suggest that the modification of NPWT provides various advantages and leads to excellent clinical outcomes in the treatment of severe deep fascial space infections. Future studies will focus on the safety verification of portable usage and the cost effectiveness analysis of NPWT.Keywords: negative pressure wound therapy, severe deep fascial space infections, modified usage, head and neck

Zhifeng Zhao, 1,* Miaomiao Liu, 1,* Yuanyuan Zhang, 1 Yingran Liang, 1 Donglai Ma, 1 Hongfang Wang, 1 Zhihong Ma, 2 Shengjiang Guan, 2 Zhonglin Wu, 3 Xi Chu, 3 Yue Lin, 2 Li Chu 1, 4 1School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, People’s Republic of China; 2School of Basic Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, People’s Republic of China; 3The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, People’s Republic of China; 4Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns, Shijiazhuang 050200, Hebei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li ChuSchool of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, People’s Republic of ChinaEmail chuli0614@126.comYue LinSchool of Basic Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, People’s Republic of ChinaTel/Fax +86 311 89926718Email yuelin8992@126.comPurpose: Monoammonium glycyrrhizinate (MAG) is an aglycone of glycyrrhizin that is found in licorice and is often used clinically as an injection to treat liver diseases. However, the effect of MAG injection on cardiac function and its possible cellular mechanisms remain unclear. We explored the protective effects of MAG against myocardial ischemic injury (MII) induced by isoproterenol (ISO), as well as the cellular mechanisms via molecular biology techniques and patch-clamp recording.Methods: A rat model of myocardial ischemia injury was induced by administering ISO (85 mg/kg) subcutaneously for 2 consecutive days. ECG, cardiac functional parameters, CK and LDH levels, SOD and GSH activities, MDA concentration, histological myocardium inspection, mitochondria ultrastructure changes, intracellular calcium concentrations were observed. Influences of MAG on ICa-L and contraction in isolated rat myocytes were observed by the patch-clamp technique.Results: MAG reduced damage, improved cardiac morphology, inhibited oxidative stress, decreased the generation of reactive oxygen species, and decreased intracellular Ca 2+ concentration. Exposure of the rats’ ventricular myocytes to MAG resulted in a concentration-dependent reduction in L-type calcium currents (ICa-L). MAG reduced ICa-L in a consistent and time-dependent fashion with a semi-maximal prohibitive concentration of MAG of 14 μM. MAG also shifted the I-V curve of ICa-L upwards and moved the activation and inactivation curves of ICa-L to the left.Conclusion: The findings indicate that MAG injection exerts a protective influence on ISO-induced MII by inhibiting oxidative stress and regulating Ca 2+ homeostasis by ICa-L.Keywords: cardiopretection, reactive oxygen species, calcium influx, isoproterenol, calcium concentration

Fan Yang,1,2 Xu Xia Wang,1,3 Dan Ma,1,2 Qun Cui,1,2 De Hua Zheng,1,2 Xiao Can Liu,1,2 Jun Zhang1,2 1Shandong Provincial Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, Shandong Province, People’s Republic of China; 2Department of Orthodontics, School of Stomatology, Shandong University, Jinan, Shandong Province, People’s Republic of China; 3Department of Oral and Maxillofacial Surgery, School of Stomatology, Shandong University, Jinan, Shandong Province, People’s Republic of ChinaCorrespondence: Jun ZhangDepartment of Orthodontics, School of Stomatology, Shandong University, 44 Wenhua West Road, Jinan 250012, People’s Republic of ChinaTel +86 139 5310 9816Fax +86-531-88565657Email zhangj@sdu.edu.cnPurpose: The aim of this study was to investigate the effects of triptolide on the tooth movement and root resorption in rats during orthodontic treatment.Material and methods: A total of 48 male Wistar rats were divided into three groups of 16 each. The right maxillary first molars of rats were drawn mesially by closed coil nickel–titanium spring with a force of 50 g. The two experimental groups received intraperitoneal injections of triptolide for 14 days at a dose of 15 μg/kg/day and 30 μg/kg/day, respectively. The control group received vehicle injections. After 14 days, the rats were humanely killed. The amount of tooth movement was measured. Eight rats from each group were randomly chosen for analysis of the percentage of root resorption area by scanning electron microscopy. For the remaining eight rats in each group, the H&E staining, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry analysis were performed.Results: The amount of tooth movement and the ratio of root resorption area were significantly decreased in the triptolide-treated rats. The number of TRAP-positive cells was significantly lower in triptolide-treated groups. Moreover, the expression of nuclear factor kappa B ligand (RANKL) was reduced. In contrast, the expression of osteoprotegerin was significantly up-regulated. In the tension side, the expressions of runt-related transcription factor 2 and osteocalcin were significantly enhanced by triptolide injection.Conclusion: Triptolide injection could arrest orthodontic tooth movement and reduce root resorption in rats via inhibition of osteoclastogenesis. In addition, triptolide may exert a positive effect on osteoblastogenesis.Keywords: triptolide, osteoclast, scanning electron microscope, tooth movement, bone formation

Wen-Xiu Yuan,1 Xu-Xia Wang,2 De-Hua Zheng,1 Dan Ma,1 Qun Cui,1 Fan Yang,1 Jun Zhang1 1Department of Orthodontics, School and Hospital of Stomatology, Shandong University and Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong Province, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Shandong University and Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong Province, People’s Republic of ChinaCorrespondence: Jun ZhangDepartment of Orthodontics, School and Hospital of Stomatology, Shandong University and Shandong Provincial Key Laboratory of Oral Tissue Regeneration, No. 44-1 Wenhua Road West, Jinan, Shandong Province, People’s Republic of ChinaTel +86 139 5310 9816Email zhangj@sdu.edu.cnObjectives: This study was performed to evaluate the effects of muscone on the proliferation, migration and differentiation of human gingival mesenchymal stem cells (GMSCs) and to explore the relevant mechanisms.Materials and methods: We performed studies to determine the effects and mechanisms of muscone on GMSC proliferation, migration and differentiation. We conducted CCK-8, colony formation, transwell chamber, scratch wound, alkaline phosphatase (ALP) staining and activity, and alizarin red and oil red O staining assays, as well as real-time quantitative polymerase chain reaction (qRT-PCR), to ascertain the effects of muscone on GMSC proliferation, migration and differentiation in vitro. The mechanism by which muscone influences the osteogenic and adipogenic differentiation of GMSCs was elucidated by qRT-PCR and Western blotting.Results: We found that muscone significantly promoted GMSC proliferation, chemotaxis, wound healing and fat droplet formation and inhibited ALP activity and mineral deposition. Notably, we observed that the Wnt/β-catenin pathway was closely related to the ability of muscone to inhibit the osteogenic differentiation and promote the adipogenic differentiation of GMSCs. The effect of muscone on the multidirectional differentiation capacity of GMSCs was significantly reversed by the agonist lithium chloride through the Wnt/β-catenin signaling pathway.Conclusion: Muscone effectively increased the proliferation and migration, promoted the adipogenic differentiation and inhibited the osteogenic differentiation of GMSCs by inhibiting the Wnt/β-catenin signaling pathway. These results may provide a theoretical basis for the application of GMSCs and muscone in tissue engineering and regenerative medicine.Keywords: muscone, gingival mesenchymal stem cells, GMSCs, differentiation, Wnt/β-catenin signaling pathway, proliferation, chemotaxis

Hongge Liang1, Di Ma2, Yan Xu1, Jing Zhao1, Minjiang Chen1, Xiaoyan Liu1, Wei Zhong1, Junling Li2, Mengzhao Wang1 1Department of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Department of Oncology, Chinese Academy of Medical Sciences Cancer Institute and Hospital, Beijing, People’s Republic of ChinaCorrespondence: Wei ZhongDepartment of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, No. 1 Shuanfuyuan Wangfujing Dongcheng District, Beijing 100730, People’s Republic of ChinaTel +86 0 106 915 5039Email zw_pumch@126.comMengzhao WangDepartment of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, No. 1 Shuanfuyuan Wangfujing Dongcheng District, Beijing 100730, People’s Republic of ChinaTel +86 0 106 915 5154Email mengzhaowang@sina.comBackground: Targeted therapy is an important treatment for advanced non-small cell lung cancer (NSCLC) patients with specific genetic mutations, crizotinib can prolong survival in advanced NSCLC patients with echinoderm microtubule-associated protein-like 4–anaplastic lymphoma kinase (EML4-ALK) rearrangement. We performed a retrospective analysis to investigate the association between the lactate dehydrogenase (LDH) levels and progression-free survival (PFS) in patients with EML4-ALK rearrangement NSCLC receiving treatment with crizotinib.Methods: Advanced (stage IIIb–IV) NSCLC patients with EML4-ALK rearrangement receiving treatment with crizotinib were enrolled between January 2007 and January 2016 at Peking Union Medical College and Cancer Hospital Chinese Academy of Medical Sciences.Results: Overall, 212 patients were enrolled. Kaplan–Meier univariate analysis showed that elevated pre-treatment LDH level (7.9 vs 14.1 months, HR =1.251, CI: 1.008–1.553, P=0.004) was significantly associated with shorter PFS, while the post-treatment mean-LDH level (13.3 vs 14.3 months, HR=1.439, 95% CI: 0.994–2.082, P=0.970) was not significantly associated with PFS. Cox proportional hazards model also identified that pre-treatment LDH level (HR=2.085, 95% CI: 1.150–3.781, P=0.016) was associated with the PFS. Logistic regression analysis showed that post-treatment LDH level was associated with creatine kinase (OR=6.712, 95% CI 3.395–13.273, P250 U/L) was significantly associated with shorter PFS in patients with EML4-ALK rearrangement NSCLC. Post-treatment elevated serum LDH level was not significantly associated with PFS, which related to adverse events including muscle damage and anemia.Keywords: non-small cell lung cancer, lactate dehydrogenase, crizotinib, echinoderm microtubule-associated protein-like 4–anaplastic lymphoma kinase, progression-free survival

De-Hua Zheng,1 Xu-Xia Wang,2 Dan Ma,1 Li-Na Zhang,3 Qing-Fang Qiao,1 Jun Zhang41Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, Shandong Province, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, School of Dentistry, Shandong University, Jinan, Shandong Province, People’s Republic of China; 3Department of Orthodontics, Liaocheng People’s Hospital, Liaocheng, Shandong Province, People’s Republic of China; 4Department of Orthodontics, School of Stomatology, Shandong University, Jinan, Shandong Province, People’s Republic of ChinaObjectives: The aim of this study is to examine the roles of erythropoietin (EPO) in regulating proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) and analyze the underlying signaling of these processes.Materials and methods: PDLSCs were isolated and characterized. The PDLSCs were transfected with β-catenin shRNA. qRT-PCR and Western blot analysis were used to examine the osteogenic effects of EPO on the expression of osteogenic-related genes and protein (Runx2, OCN and Osterix) in PDLSCs. Alizarin Red-S staining was used to detect mineralized nodule formation. In addition, the relationship between the Wnt/β-catenin pathway and the effect of EPO on the osteogenesis of PDLSCs was investigated.Results: The results suggested that EPO exerts positive osteogenic effects on PDLSCs. The results showed that EPO decreased the growth of PDLSCs slightly and increased alkaline phosphatase activity and calcium deposition in a dose-dependent manner. The expression of Runx2, Osterix and OCN was increased after EPO administration. EPO increases β-catenin and Cyclin D1 in PDLSCs. After transfected with β-catenin shRNA, the osteogenic effect of EPO on PDLSCs was attenuated.Conclusion: EPO promotes osteogenic differentiation of PDLSCs. The underlying mechanism may be activating Wnt/β-catenin signaling pathway.Keywords: erythropoietin, periodontal ligament stem cell, osteogenesis, Wnt/β-catenin

Yuanyuan Han,1,2 Xuxia Wang,2,3 Dan Ma,1,2 Xiaoxiao Wu,1,2 Panpan Yang,1,2 Jun Zhang1,2 1Department of Orthodontics, Faculty of Stomatology, Shandong University, Jinan, 2Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, 3Department of Oral and Maxillofacial Surgery, Faculty of Stomatology, Shandong University, Jinan, China Objectives: This study was performed to investigate the effects and mechanism of ipriflavone (IP) on the proliferation and osteoblastic differentiation of periodontal ligament cells in vitro and periodontal tissue remodeling following orthodontic tooth movement (OTM) in vivo. Materials and methods: Human periodontal ligament cells (hPDLCs) were cultured in vitro and cell counting kit-8, alkaline phosphatase (ALP) activity assay, plate clone formation assay, and alizarin red staining were used to test proliferation and osteogenic differentiation of hPDLCs. What is more, the expression of ALP, Runx2, and GPR30 was examined by real-time polymerase chain reaction and Western blot. To find out if PI3K/AKT signaling pathway was involved in the process, AKT and p-AKT were examined by Western blot. LY294002 (PI3K signaling pathway inhibitor) and small interfering RNA targeting GPR30 mRNA (siGPR30) were used to verify the function of GPR30-mediated PI3K/AKT pathway in this process. Twenty-four male Wistar rats were randomized into 2 groups, the control group with force application and the IP group with force application plus IP. Morphological changes in the periodontal tissue between roots of teeth were investigated using hematoxylin and eosin (HE) staining and bone morphogenetic protein-2 was detected to assess bone remodeling by immunohistochemical staining. Results: In vitro, 10-7 M IP was selected significantly promoting proliferation, ALP activity, colony forming efficiency, and mineral deposition (P