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Tao Zhang,1 Beiyin Lu,1 Zhongheng Yan,2 Xiaojun Huang,3 Wei Lu3,4 1Department of Health Policy and Management, Hangzhou Normal University, Hangzhou, People’s Republic of China; 2Office of the Vice President, Central Hospital of Hainan Province Western, Haikou, People’s Republic of China; 3School of Management, Hainan Medical University, Haikou, People’s Republic of China; 4Department of Science and Education, Hainan Women and Children’s Health Care Centre, Haikou, People’s Republic of ChinaCorrespondence: Wei Lu, School of Management, Hainan Medical University, No. 3 Xueyuan Road, Longhua District, Haikou, 5711993, People’s Republic of China, Tel +86 15971499800, Fax +8602767813016, Email hy0205044@hainmc.edu.cnPurpose: Based on the diagnosis-related groups payment, China developed an innovative episode-based payment scheme, called “payment method by disease types with point counting”, to control health expenditures inflation. This study aimed to investigate the impacts of this new payment method on volume, expenditures, and efficiency in Chinese public hospitals.Methods: The study sample consisted of 7 tertiary hospitals and 14 secondary hospitals in Jinhua (intervention group) and 4 tertiary hospitals and 14 secondary hospitals in Taizhou (control group). Monthly data points were collected for each sampled hospital from June 2016 to June 2019 using a self-administered questionnaire with impact evaluation indicators. Controlled interrupted time-series analysis was employed to estimate the effect of the new payment method.Results: The significant slowing trends in inpatient expenditures per visit (tertiary hospitals: β7=− 123.16, p=0.042; secondary hospitals: β7=− 89.24, p=0.021) and out-of-pocket payments (tertiary hospitals: β7=− 4.18, p=0.027; secondary hospitals: β7=− 4.87, p=0.019) were observed after policy intervention. However, outpatient expenditures per visit in tertiary (β7=1.67, p=0.018) and secondary hospitals (β7=1.24, p=0.003) rose faster with the new payment method. Additionally, payment reform also caused an increase in the number of inpatient visits (β7=100.01, p=0.038) and reduced the length of stay (β7=− 0.10, p=0.036) in tertiary hospitals.Conclusion: The introduction of payment method by disease types with point counting causes the cost containment for inpatient care, whereas the increase in outpatient expenditures. The findings suggest this new payment scheme has the potential for rollout in other areas, but the cost-shifting from the inpatient to outpatient setting should be prevented.Keywords: episode-based payment, expenditures, public hospitals, controlled interrupted time series, China

Meng Zhang,1,2,&ast; Zhao Wang,2,&ast; Jiaxi Wang,1 Hairong Lv,2 Xia Xiao,2 Wenyi Lu,2 Xin Jin,2 Juanxia Meng,2 Yedi Pu,2 MingFeng Zhao2 1First Central Clinical College, Tianjin Medical University, Tianjin, 300192, People’s Republic of China; 2Department of Hematology, Tianjin First Central Hospital, Tianjin, 300192, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: MingFeng Zhao, Department of Hematology, Tianjin First Central Hospital, No. 24, Fukang Road, Nankai District, Tianjin, 300192, People’s Republic of China, Tel +16622082788, Email mingfengzhao@sina.comBackground: It was crucial to use empirical antibiotics in febrile neutropenia (FN) patients. However, most patients still died from infection due to poor efficacy. Metagenomic next-generation sequencing (mNGS) is a rapid microbiological diagnostic method. The value of mNGS in patients with FN remains to be studied, especially after empiric antibiotic treatment.Methods: We retrospectively analyzed the differences between mNGS and the traditional methods in 192 patients with hematological malignancies who have received empiric antibiotic treatment. Samples were collected when patient had chills or half an hour before peak body temperature. And we compared the differences between FN and non-FN patients, mainly including types of pathogens and the diagnostic value of different pathogens.Results: Despite receiving empirical treatment, the pathogen detection rate of mNGS was significantly higher than the traditional method (80.21% vs 25.00%, P< 0.001). And it has obvious advantages in detecting mixed pathogens infection (80.21% vs 4.17%, P< 0.001). Then, we found that mNGS saw more pathogens in the FN than in the non-FN group, especially fungus. 21/33 (63.63%) of FN patients was diagnosed with fungal infections. The fungal detection rate in FN was significantly higher than non-FN group (32.35% vs 12.22%, P=0.001). Besides, the sensitivity of mNGS was higher than the traditional methods in both FN and non-FN group (P< 0.001), but no significant difference in specificity (P> 0.05). In the FN group, empiric antibiotic treatment of 46/102 (45.10%) patients did not treat all the pathogens detected by mNGS. After adjusting the antimicrobial regimen according to the results of mNGS, the effective rate at 72 hours and 7 days was 22/46 (47.83%) and 24/102 (52.17%), respectively.Conclusion: mNGS had a significant impact on the diagnosis of infection and the second-line antimicrobial therapy in FN. mNGS plays a more important role in FN patients, especially in the diagnosis of fungal infections.Purpose: Firstly, we compared the difference between mNGS and the traditional methods in the diagnosis of infection. Secondly, we assessed the value of mNGS in FN patients by comparing it with non-FN patients, including types of pathogens and the diagnostic value of different pathogens. In order to show that mNGS plays a more important role in FN.Keywords: metagenomic next-generation sequencing, febrile neutropenia, infection, fever, empiric antibiotic treatment

Wenbin Lu,1,&ast; Keqin Zhang,1,2,&ast; Xinning Chang,1 Xiya Yu,1 Jinjun Bian1 1Faculty of Anesthesiology, Changhai Hospital, Naval Medical University/Second Military Medical University, PLA, Shanghai, 200433, People’s Republic of China; 2Department of Anesthesiology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200437, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiya Yu; Jinjun Bian, Faculty of Anesthesiology, Changhai Hospital, Naval Medical Unversity, 168, Changhai Road, Shanghai, 200433, People’s Republic of China, Tel/Fax +86-021-31161886 ; +86-021-31161841, Email yuxiyash@163.com; jinjunbicu@163.comPurpose: Postoperative cognitive decline (POCD) is highly prevalent in elderly patients who received surgery. The systemic immune-inflammation index (SII) has been shown to be an independent predictor of many diseases associated with inflammation, but the relationship between the SII and POCD is unknown. We aimed to investigate the association between POCD and SII levels to examine the potential of SII in predicting POCD in elderly patients.Patients and Methods: The present study was carried out among elderly patients who underwent elective orthopedics operation in our hospital, and SII, neutrophil to lymphocyte ratio (NLR), monocyte to lymphocyte ratio (MLR) were calculated from the admission blood sample. POCD was measured by Mini-mental State Examination (MMSE) in elderly patients. The association between SII levels and POCD was analyzed by binary logistic regression analysis.Results: Finally, 19 (25%) of 76 patients were diagnosed with POCD. Compared with Non-POCD patients, POCD patients showed significantly higher levels of NLR, MLR, SII, especially SII at admission. SII was independently associated with the occurrence of POCD through the logistic regression analysis. Receiver operating characteristic curve analysis indicated that preoperative SII was a significant predictor for POCD, and the area under the curve was 0.909.Conclusion: Our data suggest that preoperative NLR, MLR, SII levels in the blood are related to the occurrence of POCD. Preoperative SII level is a prognostic biomarker of POCD in elderly patients after orthopedics operation. More clinical studies are needed to further verify the value of SII in POCD.Keywords: systemic immune-inflammation index, postoperative cognitive decline, geriatric, neutrophil to lymphocyte ratio, monocyte to lymphocyte ratio

Zeyu Wang,1,&ast; Yonghe Zhou,2,3,&ast; Pengzhi Yu,2,3 Yonggang Liu,3,4 Mei Mei,5 Zhuo Bian,2 Wei Shao,2 Jinxia Lv,2 Xin Li,2 Wei Lu,1,3,6 Liang Xu3,6 1Department of Hepatobiliary Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin’s Clinical Research Center for Cancer, Tianjin, 300060, People’s Republic of China; 2Ultrasound department, Tianjin Second People’s Hospital, Tianjin, 300192, People’s Republic of China; 3Tianjin Research Institute of Liver Diseases, Tianjin, 300192, People’s Republic of China; 4Pathology Department, Tianjin Second People’s Hospital, Tianjin, 300192, People’s Republic of China; 5Department of Gastroenterology, Tianjin Haihe Hospital, Tianjin, 300350, People’s Republic of China; 6Department of Hepatology, Tianjin Second People’s Hospital, Tianjin, 300192, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Lu; Liang Xu, Email luwei1966@126.com; xuyangliang2004@sina.comBackground: At present, there is a lack of cheap, effective and convenient detection methods for hepatitis B-related liver fibrosis, especially in the developing area.Aim: To evaluate the non-invasive methods for the significant and advanced fibrosis stage in chronic hepatitis B virus (HBV) patients in basic hospitals and to assess their diagnostic utility.Methods: The study included 436 consecutive naive HBV individuals who had their livers biopsied. They were examined in one week using aspartate aminotransferase-to-aspartate aminotransferase ratio (AAR), age-platelet index (API), aspartate aminotransferase-to-platelet ratio index (APRI), fibrosis-4 (FIB-4), Forns, gamma-glutamyl transpeptidase-to-platelet ratio (GPR), S-index and transient elastography (TE). Scheuer scoring system was used to determine the histologic fibrosis grades (S0–S4). The diagnostic effectiveness was assessed using AUROCs and the DeLong test, both of which were based on statistical comparisons.Results: For both substantial (≧S2) and advanced (≧S3) fibrosis phases, TE had good diagnostic performance in determining the hepatic fibrosis. Similar diagnostic performance was shown with Forns and S-index when it came to detecting fibrosis stages lower than S3. One model’s diagnostic value was not significantly improved by combining serum models. Correlation coefficients between clinical features and fibrosis phases were greatest for Forns (r = 0.397), S-index (r = 0.382) and TE (r = 0.535) when compared to other variables.Conclusion: This investigation showed that Forns and S-index may be helpful strategies for detecting advanced fibrosis in HBV patients admitted to community hospitals.Keywords: hepatitis B virus, non-invasive, basic hospital, transient elastography, Forns, S-index

Jingxia Sun, Wensheng Lu Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Jingxia Sun, Jianhao Huang, Wensheng Lu Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Jiaxia Han, Wensheng Lu Department of Endocrinology, People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Qiu Wang, Xiuxian Huang, Wensheng Lu Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of ChinaCorrespondence: Wensheng Lu, Email Lwswxqz@163.com

Wenjun Lu,1,&ast; Quedan Qiu,2,&ast; Keda Chen,2 Rongqing Zhao,2 Qingcao Li,2 Qiaoping Wu2 1Intensive Care Units of Ningbo Medical Centre Lihuili Hospital, Ningbo University, Ningbo, Zhejiang Province, People’s Republic of China; 2Clinical Laboratory of Ningbo Medical Centre Lihuili Hospital, Ningbo University, Ningbo, Zhejiang Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qingcao Li; Qiaoping Wu, Tel +86-574-55835786, Fax +86-574-55835781, Email lqc_lab@163.com; lhlyywqp@163.comBackground: Integrons are the main mode of horizontal transmission of drug-resistance genes and are closely related to drug resistance in clinical bacteria. In this study, the distributions of class 1, 2, and 3 integron gene cassettes were investigated in 150 Proteus mirabilis (P. mirabilis) isolates from patients, and molecular characterization of functional class 2 integrons was further analyzed.Methods: Class 1, 2, and 3 integrons were screened by polymerase chain reaction (PCR) in 150 clinical P. mirabilis isolates. The variable regions of the integrons were determined by restriction analysis and sequencing. Internal stop codons mutations in class 2 integrons and their common promoters were also determined by sequencing. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) was used to analyze the phylogenetic relations of class 2 integron-positive isolates.Results: Class 1 integrons were detected in 69 (46%) of 150 P. mirabilis isolates, and six different gene cassette arrays were detected, with the most prevalent being dfrA32-aadA2. Class 2 integrons were detected in 61 (40.7%) of 150 P. mirabilis isolates, and three different gene cassette arrays were detected, including sat2-aadA1, which was detected for the first time in a class 2 integron. Nearly similar ERIC-PCR fingerprinting patterns were detected in 45 (73.8%) of 61 class 2 integron-positive isolates. The functional class 2 integron was detected in three P. mirabilis isolates having the same gene cassette, dfrA1-sat2-aadA1, in the variable region and four novel open reading frames with unknown functions. Same PintI2 and Pc promoters were detected in these three functional class 2 integron isolates, as was found in other class 2 integron isolates. However, these three strains did not totally show identical homology and drug sensitivity.Conclusion: Although functional class 2 integrons have low distribution and relatively conserved molecular characteristics, they can still form clinical dissemination and drug resistance expression.Keywords: Proteus mirabilis, integrons, premature stop codon, promoter regions, antibacterial drug resistance

Na Wang,1 Minghua Zhan,1,2 Jianhua Liu,3 Yao Wang,4 Yongwang Hou,5 Caiqing Li,1 Jia Li,1 Xuying Han,1 Jinlu Liu,1 Yong Chen,6 Jingjing Fan,6 Jianhua Tang,7 Wenhua Lu,8 Xinran Zhong,1 Zhihua Zhang,3 Wei Zhang1,4 1Microbiology Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 2Clinical Laboratory, Peking University People’s Hospital, Beijing, 100730, People’s Republic of China; 3Respiratory Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 4Clinical Laboratory, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 5Clinical Laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 6Infectious Disease Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 7Clinical Pharmacy Department, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, People’s Republic of China; 8Dental Department, Beijing Fengtai Tieying Community Health Service Center, Beijing, 100730, People’s Republic of ChinaCorrespondence: Wei Zhang; Zhihua Zhang, Tel +86-15369318318, Email 15369318318@163.com; zzh19641229@163.comPurpose: In this study, we analyzed the clinical distribution and drug resistance of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains, the minimum inhibitory concentrations (MIC), MIC50 and MIC90, and geographical distribution in Hebei Province, China. We aimed to provide epidemiological research data, formulate appropriate combined treatment schemes, reasonably select antibiotics, and standardize nosocomial infection control schemes.Patients and Methods: A total of 6328 strains of CRKP were collected from 2017 to 2019. The MIC was determined for the drug sensitivity test, and the experimental data were statistically analyzed using WHONET5.6.Results: The detection rate of CRKP increased annually from 13.4% in 2017 to 14.5% in 2018, and 14.6% in 2019. The ratio of males to females was approximately 2:1; 53.6% were elderly, 39% were adults, 4.8% were minors, and 2.5% were newborns. The specimens collected were mainly sputum (70.9%). The resistance rate of CRKP to carbapenems and other β-lactam antibiotics was found to be increasing, with resistance rates generally greater than 90%. The resistance rate to aminoglycoside antibiotics decreased yearly to approximately 50%, and the resistance rate to quinolones remained unchanged at approximately 80%. From 2017 to 2019, the resistance rate of CRKP in Hebei Province to various antibiotics was high, and the resistance rate to β-lactam antibiotics increased each year.Conclusion: The situation of CRKP resistance is severe in Hebei Province, China. The resistance rate to most antibiotics is very high and shows an upward trend. Among them, the resistance rate to polymyxin is low; however, few resistant strains do exist. MIC50 and MIC90 are higher than their MICs. It mainly causes lung infection in elderly men. This study is helpful to improve the diagnosis, treatment, and prevention of CRKP infection in our province.Keywords: carbapenems, clinical distribution, drug resistance, minimum inhibitory concentration, geographic distribution, Klebsiella pneumoniae

Wang Li,1,2,&ast; Mei Mei,1,3– 5,&ast; Tian Liu,1,&ast; ShuWen Zhang,1 ZeYu Wang,1 YuHong Suo,1 Shuai Wang,1 Yang Liu,1 NingNing Zhang,1 Wei Lu1 1Department of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University, Tianjin, 300060, People’s Republic of China; 2NHC Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin, 300134, People’s Republic of China; 3Graduate School of Tianjin Medical University, Tianjin, 300070, People’s Republic of China; 4Tianjin First Central Hospital, Tianjin, 300192, People’s Republic of China; 5Department of Gastroenterology, Tianjin Haihe Hospital, Tianjin, 300350, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Lu; NingNing ZhangDepartment of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University, Tianjin, 300060, People’s Republic of ChinaEmail mail4luwei@163.com; mail4ningning@163.comBackground: Hepatocellular carcinoma has been identified to be among the most prevalent malignancies in the world and has an unfavorable prognosis. Immune checkpoints perform an essential function in many biological processes and are associated with the survival of cancer patients. The function of immune checkpoints remains unknown.Methods: We used bioinformatic methods to examine the prognostic value of immune checkpoints and the corresponding link to immune infiltration in HCC. qRT-PCR was used to validate the expression of immune checkpoints and their prognostic significance in HCC.Results: The level of mRNA of SIGLEC15, PDCD1LG2, LAG3, PDCD1, CTLA4 as well as PDCD1LG2 was increased in HCC tissues as opposed to liver tissues. Immune checkpoints were shown to participate in the activation of the apoptotic pathway in HCC patients. The elevated expression of PDCD1 and PDCD1LG2 were shown to have a favorable recurrence-free survival (RFS), progression-free survival (PFS), disease-specific survival (DSS), and overall survival (OS). PDCD1, PDCD1LG2, and pT stage were independent variables that affect the HCC patients’ prognoses as revealed by the multivariate and univariate analyses. A prediction nomogram indicated that the calibration plots for OS rates over three and five years had a stronger predictive performance in the TCGA HCC cohort in contrast with an ideal model. Positive correlations were observed between the PDCD1 and PDCD1LG2 expression and immune biomarkers, immune cells, chemokine receptors, as well as chemokines.Conclusion: The present research performed a thorough examination of the prognostic significance of immune checkpoints in HCC and its correlation with immune infiltration, which suggested that PDCD1 and PDCD1LG2 were prognostic biomarkers in HCC and related to the immune infiltration.Keywords: HCC, PDCD1, PDCD1LG2, immune checkpoints, prognosis biomarker

Samuel James Brake,1 Mathew Suji Eapen,1 Kielan Darcy McAlinden,1 James Markos,1,2 Greg Haug,1,2 Josie Larby,1,2 Collin Chia,1,2 Ashutosh Hardikar,1,3 Gurpreet Kaur Singhera,4,5 Tillie L Hackett,4,5 Wenying Lu,1 Sukhwinder Singh Sohal1 1Respiratory Translational Research Group, Department of Laboratory Medicine, School of Health Sciences, College of Health and Medicine, University of Tasmania, Launceston, TAS, 7248, Australia; 2Department of Respiratory Medicine, Launceston General Hospital, Launceston, TAS, 7250, Australia; 3Department of Cardiothoracic Surgery, Royal Hobart Hospital, Hobart, TAS, 7000, Australia; 4Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, BC, Canada; 5Department of Medicine, University of British Columbia Centre for Heart Lung Innovation, St. Paul’s Hospital, Vancouver, BC, CanadaCorrespondence: Sukhwinder Singh SohalRespiratory Translational Research Group, Department of Laboratory Medicine, School of Health Sciences, College of Health and Medicine, University of Tasmania, Locked Bag – 1322, Newnham Drive, Launceston, TAS, 7248, AustraliaTel +61 3 6324 5434Email sssohal@utas.edu.auBackground: Smokers and patients with COPD are highly susceptible to SARS-CoV-2 infection, leading to severe COVID-19.Methods: This cross-sectional study involved resected lung tissues from 16 patients with GOLD stage I or II COPD; of which 8 were current smokers COPD (COPD-CS), and 8 ex-smokers COPD (COPD-ES), 7 normal lung function smokers (NLFS), 9 patients with small airways disease (SAD), and 10 were never-smoking normal controls (NC). Immunostaining for ACE2, Furin, and TMPRSS2 was performed and analysed for percent expression in small airway epithelium (SAE) and counts for positively and negatively stained type 2 pneumocytes and alveolar macrophages (AMs) were done using Image ProPlus V7.0. Furthermore, primary small airway epithelial cells (pSAEC) were analysed by immunofluorescence after exposure to cigarette smoke extract (CSE).Results: ACE2, Furin, and TMPRSS2 expression significantly increased in SAE and type 2 pneumocytes in all the subjects (except Furin for NLFS) compared to NC (p < 0.001). Similar significance was observed for ACE2 positive AM (p < 0.002), except COPD-ES, which decreased in ACE2 positive AMs (p < 0.003). Total type 2 pneumocytes and AMs significantly increased in the pathological groups compared to NC (p < 0.01), except SAD (p = 0.08). However, AMs are significantly reduced in COPD-ES (p < 0.003). Significant changes were observed for tissue co-expression of Furin and TMPRSS2 with ACE2 in SAE, type 2 pneumocytes and AMs. These markers also negatively correlated with lung function parameters, such as FEV1/FVC % predicted, FEF25-75%, DLCO% predicted. A strong co-localisation and expression for ACE2 (p < 0.0001), Furin (p < 0.01), and TMPRSS2 (p < 0.0001) was observed in pSAEC treated with 1% CSE than controls.Discussion: The increased expression of ACE2, TMPRSS2 and Furin, in the SAE, type 2 pneumocytes and AMs of smokers and COPD are detrimental to lung function and proves that these patient groups could be more susceptible to severe COVID-19 infection. Increased type 2 pneumocytes suggest that these patients are vulnerable to developing post-COVID-19 interstitial pulmonary fibrosis or fibrosis in general. There could be a silently developing interstitial pathology in smokers and patients with COPD. This is the first comprehensive study to report such changes.Keywords: COVID-19, SARS-CoV-2, smoking, COPD, ACE2, type 2 pneumocytes, alveolar macrophages, epithelium

Haoming Chen,&ast; Jianmang Yu,&ast; Lihua Hang,&ast; Shuai Li, Weikang Lu, Zhenkai Xu Department of Anesthesiology, Kunshan First People’s Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, 215300, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lihua HangDepartment of Anesthesiology, Kunshan First People’s Hospital Affiliated to Jiangsu University, Suzhou, Jiangsu Province, Peoples Republic of ChinaTel +86-18013273169Email zjhanglihua@foxmail.comIntroduction: Bone cancer pain (BCP) seriously affects the quality of life of patients with advanced cancer, but effective treatment methods are lacking. This study mainly investigates the role of EZH2 in a well-established BCP model induced by Walker 256 breast cancer cells in rats.Methods: Female Sprague–Dawley rats of the same age weighing approximately 160 g were selected for the experiment. The BCP model was established by injecting inactivated Walker 256 breast cancer cells into the tibia. von Frey filaments were used to measure the paw withdrawal threshold, and bone destruction in the rat was observed using x-ray. The spinal EZH2 and H3K27Tm levels were measured using Western blotting and RT–qPCR analysis. Intrathecal injection of an EZH2 inhibitor was performed to examine the role of EZH2 in trigeminal BCP.Results: Experimental results showed that injecting Walker 256 breast cancer cells into the tibia induced bone cancer pain. Spinal EZH2 and H3K27Tm levels were significantly increased over time in BCP rats. An intrathecal injection of 3-deazaneplanocin A (DZNep), a selective EZH2 inhibitor, downregulated the expression of EZH2 and attenuate the BCP-induced mechanical allodynia state.Conclusion: Intrathecal injection of DZNep relieve bone cancer pain in rats. EZH2 expressed in spinal cord tissue may be involved in the process of bone cancer pain in rats.Keywords: enhancer of zeste homologue 2, bone cancer pain, histone methyltransferase, 3 deazaneplanocin A, intrathecal injection

Lingwen Ying,1,&ast; Yong Zhang,2,&ast; Jun Yin,1,&ast; Yufei Wang,1 Wei Lu,1 Wei Zhu,1 Yuqian Bao,1 Jian Zhou1 1Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital; Shanghai Clinical Center for Diabetes; Shanghai Diabetes Institute; Shanghai Key Laboratory of Diabetes Mellitus, Shanghai, 200233, People’s Republic of China; 2Department of Immunology and Microbiology, Shanghai Jiao Tong University, School of Medicine, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jian ZhouDepartment of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, 600 Yishan Road, Shanghai, 200233, People’s Republic of ChinaTel +86-21-64369181Fax +86-21-64368031Email zhoujian@sjtu.edu.cnPurpose: We aimed to explore the immunological characteristics and cytokine profile of the initial stage of type 1 diabetes.Patients and Methods: In total, 123 age- and sex-matched subjects with newly diagnosed classic type 1 diabetes mellitus (T1ADM), fulminant type 1 diabetes mellitus (FT1DM), and normal glucose tolerance (NGT) were enrolled. Serum cytokine levels were measured using Milliplex MAP multifactor detection.Results: There was a significant increase in the levels of transforming growth factor β (TGFβ 1) and TGFβ 2 and decrease in programmed death-1 (PD-1), PD ligand 1 (PD-L1), pro-inflammatory cytokines, and anti-inflammatory cytokines in type 1 diabetes patients compared with the NGT subjects (all P < 0.05). There was no significant difference in C-reactive protein (CRP) and blood routine indicators between the two groups. Type 1 diabetes was further divided into T1ADM and FT1DM subgroups. FT1DM patients had much higher CRP levels than T1ADM patients (4.90 [0.95– 26.05] mg/L vs 0.39 [0.20– 0.74] mg/L, P < 0.01). Blood routine results showed that the number of leukocytes was significantly increased in FT1DM compared with that in T1ADM (9.2 [5.1– 18.8] × 109 cells/L vs 5.4 [4.5– 6.7] × 109 cells/L, P < 0.01). In FT1DM patients, neutrophil% was increased, and lymphocyte% was declined significantly, compared with that in T1ADM patients (neutrophil%: 80.2 [59.2– 85.2]% vs 59.5 [54.8– 64.0]%; lymphocyte%: 18.3 [10.1– 32.3]% vs 32.6 [26.8– 35.9]%; both P < 0.01). However, there was no difference between FT1DM and T1ADM in cytokine profile except for the decrease in CTLA-4 in T1ADM (P < 0.05).Conclusion: Compared with T1ADM, CRP and leukocytes’ levels were increased significantly in FT1DM, with an increase in neutrophil% and decline in lymphocyte%, suggesting that FT1DM may have more abrupt onset and occur as a more serious subtype of type 1 diabetes mellitus.Keywords: cytokine profile, fulminant type 1 diabetes mellitus, immune response, multifactor detection, type 1 diabetes mellitus

Hui Li,1,2,&ast; Xiao-Lei Fan,1,2,&ast; Yi-Nan Wang,1,2 Wei Lu,1,2 Haoyi Wang,1,2 Runzhi Liao,1,2 Min Zeng,1,2 Jun-Xiao Yang,1,2 Yihe Hu,1,2 Jie Xie1,2 1Department of Orthopedics, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 2Hunan Engineering Research Center of Biomedical Metal and Ceramic Implants, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jie Xie; Yihe HuDepartment of Orthopedics, Xiangya Hospital, Central South University, #87 Xiangya Road, Changsha, Hunan, 410008, People’s Republic of ChinaTel +86– 731– 89753006; +86– 731– 89753706Email dr_xiejie@163.com; csuhuyihe@163.comPurpose: Wear debris particle-induced periprosthetic osteolysis is a severe complication of total joint replacement that results in aseptic loosening and subsequent arthroplasty failure. No effective therapeutic agents or drugs have been approved to prevent or treat osteolysis; thus, revision surgery is often needed. Extracellular vesicles (EVs) are vital nanosized regulators of intercellular communication that can be directly applied to promote tissue repair and regeneration. In this study, we assessed the therapeutic potential of EVs from human urine-derived stem cells (USCs) (USC-EVs) in preventing ultrahigh-molecular-weight polyethylene (UHMWPE) particle-induced osteolysis.Methods: USCs were characterized by measuring induced multipotent differentiation and flow cytometry. USC-EVs were isolated and characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS) and Western blotting. RAW264.7 cells and bone marrow mesenchymal stem cells (BMSCs) were cultured with USC-EVs to verify osteoclast differentiation and osteoblast formation, respectively, in vitro. The effects of USC-EVs were investigated on a UHMWPE particle-induced murine calvarial osteolysis model by assessing bone mass, the inflammatory reaction, and osteoblast and osteoclast formation.Results: USCs differentiated into osteogenic, adipogenic and chondrogenic cells in vitro and were positive for CD44, CD73, CD29 and CD90 but negative for CD34 and CD45. USC-EVs exhibited a cup-like morphology with a double-layered membrane structure and were positive for CD63 and TSG101 and negative for calnexin. In vitro, USC-EVs promoted the osteogenic differentiation of BMSCs and reduced proinflammatory factor production and osteoclastic activity in RAW264.7 cells. In vivo, local injection of USC-EVs around the central sites of the calvaria decreased inflammatory cytokine generation and osteolysis compared with the control groups and significantly increased bone formation.Conclusion: Based on our findings, USC-EVs prevent UHMWPE particle-induced osteolysis by decreasing inflammation, suppressing bone resorption and promoting bone formation.Keywords: extracellular vesicles, urine-derived stem cells, UHMWPE, wear particle-induced osteolysis, anti-inflammatory

Min Xu,1,&ast; Dingchao Xiang,2,&ast; Wenhua Wang,1 Long Chen,1 Wei Lu,1 Feng Cheng3 1Department of Neurosurgery, Kunshan Hospital of Traditional Chinese Medicine, Kunshan Affiliated Hospital of Nanjing University of Chinese Medicine, Kunshan City, Jiangsu Province, 215300, People’s Republic of China; 2Department of Neurosurgery, Wuxi clinical medical school of Anhui Medical University, 904th Hospital of PLA(Taihu Hospital of Wuxi), Wuxi, 214000, People’s Republic of China; 3Department of Neurosurgery, Affiliated Kunshan Hospital of Jiangsu University, Kunshan, 215300, Jiangsu Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Feng ChengDepartment of Neurosurgery, Affiliated Kunshan Hospital of Jiangsu University, Kunshan, 215300, Jiangsu Province, People’s Republic of ChinaTel +86-13913208695Email fengchengjiangsu@163.comBackground: Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key regulator responsible for oxidative stress in brain injury. This study aimed to investigate the potential mechanism of miR-448-3p and Nrf2 in cerebral ischemia/reperfusion (I/R) injury.Methods: In vitro and in vivo cerebral I/R injury models were constructed, and Nrf2 expression levels were detected by qRT-PCR and Western blot. The potential miRNAs for Nrf2 were predicted by bioinformatic analysis. The binding interaction between miR-448-3p and Nrf2 was determined by luciferase reporter assay. The effects of miR-448-3p on neurological deficit, infarct volume, and brain water content in mice were tested. The effects of miR-448-3p on oxidative stress indicators (SOD activity, MDA content, and ROS production) were detected by commercial assay kits. The levels of HO-1 and cleaved caspase-3 were evaluated by Western blot. Cell viability was evaluated by MTT assay, and cell apoptosis was evaluated by TUNEL staining and flow cytometry.Results: Nrf2 was significantly downregulated and miR-448-3p was upregulated in cerebral I/R injury both in vivo and in vitro. MiR-448-3p downregulation efficiently attenuated brain injury and reduced oxidative stress and apoptosis. MiR-448-3p was identified to act as ceRNA of Nrf2 and negatively regulated Nrf2 expression, which was consistent with the animal studies. In addition, Nrf2 silencing obviously attenuated the neuroprotective effects of miR-448-3p inhibitor in vitro.Conclusion: MiR-448-3p participated in the regulation of cerebral I/R injury via inhibiting Nrf2.Keywords: ischemia/reperfusion, miR-448-3p, Nrf2, oxidative stress, apoptosis

Jianhao Huang,1,&ast; Weiwei Li,2,&ast; Suosu Wei,3,&ast; Xing Zhou,1 Yuechou Nong,1 Jingxia Sun,1 Zhenwei Zhai,1 Wensheng Lu1 1Department of Endocrinology and Metabolism, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of China; 2The Office of Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of China; 3Editorial Board of Chinese Journal of New Clinical Medicine, Guangxi Academy of Medical Sciences and the People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, 530021, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wensheng Lu Email lws2613676@sohu.comAim: The purpose of this study was to explore the association between estimated glomerular filtration rate (eGFR) and clinical outcomes in patients with diabetic foot osteomyelitis (DFO).Methods: This was a retrospective observational study. A total of 199 patients with DFO were recruited and divided into three groups by eGFR: normal kidney function group (eGFR ≥ 90), mildly decreased kidney function group (eGFR 60– 89) and moderately to severely decreased kidney function group (eGFR < 60). The patients were followed-up for a median of 36 months, and the study outcomes were all-cause mortality and major cardiovascular adverse events (MACE). Cox proportional hazard models were used to assess the association between eGFR and the outcomes, and a stratified analysis by sex was conducted.Results: During follow-up, all-cause mortality occurred in 51 (25.63%) patients among 199 participants, 54 (28.72%) had MACE in 188 participants and 26 (48.15%) of them died. After fully adjusting for potential confounders, compared to eGFR < 90 mL/min/1.73 m2, eGFR ≥ 90 mL/min/1.73 m2 had lower incidence of all-cause mortality (HR = 0.43, 95% CI: 0.22– 0.85; P = 0.015) and MACE (HR = 0.51, 95% CI: 0.27– 0.96; P = 0.038). Additionally, compared to eGFR < 90 mL/min/1.73 m2, eGFR ≥ 90 mL/min/1.73 m2 was independently associated with decreased risk of all-cause mortality (HR = 0.33; 95% CI 0.14– 0.76, P = 0.010) and MACE (HR = 0.27; 95% CI 0.11– 0.65, P = 0.004) in male, but not in female.Conclusion: In conclusion, decreased eGFR is a risk factor for all-cause mortality and MACE in individuals with DFO. Additionally, male with decreased eGFR had a higher risk of all-cause mortality and MACE, but female did not.Keywords: diabetic foot osteomyelitis, estimated glomerular filtration rate, prognosis

Aobo Zhuang,1 Qian Wu,1 Hanxing Tong,1,2 Yong Zhang,1,2 Weiqi Lu1,2 1Department of General Surgery, South Hospital of Zhongshan Hospital/Shanghai Public Health Clinical Center, Fudan University, Shanghai, People’s Republic of China; 2Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Weiqi Lu; Yong Zhang Email lu.weiqi@zs-hospital.sh.cn; 13681971072@163.comBackground and Objectives: Surgical treatment is still the mainstay of curative therapy for retroperitoneal liposarcoma (RLPS), but often recurs after surgical resection. We aimed to establish a nomogram for postoperative recurrence of RLPS based on the Asian population.Methods: Patients after surgical resection at the South Hospital of Zhongshan Hospital/Shanghai Public Health Clinical Center between August 2011 and December 2020 were included. The enrolled patients are randomly divided into training set and test set according to the ratio of 7:3. Prognostic factors were chosen based on Akaike Information Criterion, and the nomogram was built based on Cox regression and then internally validated through calibration plots and concordance index (C-index).Results: A total of 447 patients were included. Gender, age, presentation status, organ invasion and FNCLCC grade were used to build nomogram. The calibration plots showed that RFS predicted probabilities are identical to the actual RFS rates. The C-index of the nomogram was 0.703 (95% CI 0.623– 0.783) in the training set and 0.695 (95% CI 0.565– 0.825) in test set.Conclusion: The nomogram we established can accurately predict postoperative recurrence of RLPS patients for Asian population.Keywords: retroperitoneal soft tissue sarcoma, liposarcoma, recurrence, nomogram, surgery

Xueli Lv,1,* Xiaoxiao Li,1,* Kedi Guo,1,* Tong Li,1 Yuping Yang,1 Wensi Lu,1 Shuting Wang,1 Su Liu1,2 1Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical University, Xuzhou, Jiangsu, People’s Republic of China; 2Department of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou Medical University, Xuzhou, Jiangsu, People’s Republic of China*These authors contributed equally to this workCorrespondence: Su LiuDepartment of Anesthesiology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou Medical University, Xuzhou, Jiangsu, 221000, People’s Republic of ChinaTel +86 18118309692Fax +0516-8346-9496Email 150040009@qq.comObjective: This study aimed to explore the effects of lidocaine on postoperative quality of recovery (QoR) and immune function in patients undergoing laparoscopic radical gastrectomy.Methods: In total, 135 patients were enrolled and were equally randomized to receive low-dose lidocaine (Group LL: 1.5 mg/kg bolus followed by an infusion at 1.0 mg/kg/hour) or high-dose lidocaine (Group HL: 1.5 mg/kg bolus followed by an infusion at 2.0 mg/kg/hour) or Controls (Group C: received a volume-matched normal saline at the same rate). The primary outcome was a QoR-40 score on postoperative day (POD) 1. Secondary outcomes were a QoR-40 score on POD 3, levels of inflammatory factors (IL-6, IL-10, TNF-α) and CD4+T cells, CD8+T cells proportions, and CD4+/CD8+ cell ratios and postoperative recovery of bowel function.Results: There were no statistically significant differences in patient characteristics at baseline. The total QoR-40 scores on POD 1 in Group HL (171.4± 3.89) were higher than those in Group LL (166.20± 4.05) and in Group C (163.40± 4.38) (adjusted P< 0.001). Differences in the dimension scores of QoR-40 for pain, physical comfort, and emotional state were significant across the three groups. Lidocaine administration significantly reduced the release of IL-6, IL-10, TNF-α, and attenuated immune changes induced by trauma. Kaplan–Meier curves showed that the median time to the first exhaust and defecation were shorter in the Group HL than in Groups LL and C (1.55 days vs 2.4 days vs 2.6 days, log rank P< 0.0001; and 2.86 days vs 3.22 days vs 3.46 days, log rank P=0.002, respectively). Additionally, patients in lidocaine groups required less remifentanil consumption and experienced lower pain intensity, compared with the control group.Conclusion: Systemic lidocaine improved postoperative recovery, alleviated inflammation and immunosuppression, and accelerated the return of bowel function, and is thus, worthy of clinical application.Clinical Trials Registration: ChiCTR2000028934.Keywords: lidocaine, QoR-40, postoperative quality of life, radical gastrectomy

Xuanwei Zheng,1– 3,* Qiaolin Wang,1– 3,* Yan Luo,1– 3 Wenhua Lu,1– 3 Liping Jin,1– 3 Menglin Chen,1– 3 Wu Zhu,1– 3 Yehong Kuang1– 3 1Department of Dermatology, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 2National Clinical Research Center for Geriatric Disorders (Xiangya Hospital), Changsha, 410008, People’s Republic of China; 3Hunan Engineering Research Center of Skin Health and Disease, Hunan Key Laboratory of Skin Cancer and Psoriasis (Xiangya Hospital), Changsha, 410008, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wu Zhu; Yehong KuangXiangya Hospital, Central South University, 87 Xiangya Road, Changsha, Hunan, People’s Republic of ChinaTel +86-13319509308; +86-13574171102Email zhuwu70@hotmail.com; yh_927@126.comBackground: The seasonal patterns of psoriasis have been observed in previous studies. However, no published data indicated the risk factors associated with the seasonal variation.Purpose: This study aimed to investigate potentially related factors associated with seasonal pattern of psoriasis and provide possible implications for alleviating psoriasis in clinical practice.Patients and Methods: The retrospective study was conducted in Chinese patients with psoriasis. Demographic and clinical information were collected. Multivariable logistic regression analyses (calculating adjusted odds ratios [AORs]) were used to analyze data.Results: We continually enrolled 2270 patients (1496 males and 774 females) with psoriasis based on inclusion criteria. Disease duration (AOR=1.06, 95% CI: 1.05– 1.07), hyperlipidemia (AOR=1.77, 95% CI: 1.06– 2.98) and smoking (AOR=1.40, 95% CI: 1.17– 1.68) were significantly associated with severe psoriasis in autumn/winter. Age (AOR=0.98, 95% CI:0.97– 0.99) and occupations with more sunlight exposure (AOR=0.78, 95% CI: 0.61– 0.99) were negatively associated with the seasonal aggravation. Subgroup analysis showed that occupations with more sunlight exposure (AOR=0.64, 95% CI: 0.43– 0.94) were protective factors only in late-onset psoriasis but not early-onset, while smoking (AOR=1.39, 95% CI: 1.11– 1.74) was risk factor in the early-onset psoriasis.Conclusion: Psoriatic patients who had occupation with more sunlight exposure were less likely to report aggravation of psoriasis in autumn/winter. On the contrary, smoking and hyperlipidemia were positively associated with the seasonal aggravation. Additional prospective study is needed to identify the causality.Keywords: psoriasis, seasonal variation, weather, occupational exposure, ultraviolet, smoking

Yan Sun,1,2 Lixin Yu,1,2 Wei Gao,1,2 Jinzhen Cai,1,2 Wentao Jiang,1,2 Wei Lu,3 Yihe Liu,1,2 Hong Zheng1,2 1Organ Transplantation Center, Tianjin First Central Hospital, Tianjin, 300192, People’s Republic of China; 2Tianjin Key Laboratory for Organ Transplantation, Tianjin, People’s Republic of China; 3Liver Cancer Treatment Centre, Tianjin Medical University Cancer Institute and Hospital, Tianjin, People’s Republic of ChinaCorrespondence: Yihe Liu; Hong ZhengOrgan Transplantation Center, Tianjin First Central Hospital, No. 24 of Fukang Street, Nankai District, Tianjin, 300192, People’s Republic of ChinaTel/Fax +8602223627027Email limyihe@163.com; zhemghomgi@21cn.comObjective: This study aimed to investigate the colonization and prevalence of carbapenem-resistant Enterobacteriaceae (CRE) in pediatric liver transplant recipients and analyze the high-risk factors and prognosis of CRE infection.Methods: A prospective study involving 152 pediatric patients undergoing liver transplantation was carried out. Anal swab bacteria cultures were collected when the patients entered the intensive care unit (ICU) and when they left in order to screen for intestinal CRE colonization. The results were grouped according to the occurrence of CRE infection following surgery, and the patients were divided into two groups: a CRE infection group and a non-CRE infection group. Univariate analysis and multiple logistic regression analysis were conducted to determine the independent risk factors of CRE infection and analyze the survival rate.Results: Of the 152 pediatric liver transplant recipients enrolled in the study, there were 13 cases of postoperative CRE infection and 139 cases of non-CRE infection. The incidence of preoperative CRE infection, preoperative cytomegalovirus (CMV) infection, and preoperative sepsis in the CRE infection group was significantly higher than in the non-CRE infection group (P < 0.005). Intraoperative bleeding volume and operation times in the CRE infection group were also significantly higher than in the non-CRE infection group (P < 0.05). Furthermore, postoperative ICU treatment time, postoperative occurrence of unplanned surgery, postoperative mechanical ventilation of more than 24 hours, and the incidence of pre-ICU CRE colonization in the CRE infection group were significantly higher than in the non-CRE infection group (P < 0.05). Finally, the difference between the CRE infection group and the non-CRE infection group in six-month survival rate following surgery was significant (P < 0.001).Conclusion: The independent risk factors of CRE infection following pediatric liver transplantation include preoperative CRE infection and pre-ICU CRE colonization. CRE infection progresses quickly, with a poor prognosis and a high mortality rate. The CRE screening of anal swabs is crucial for the early detection of CRE infection.Keywords: carbapenem-resistant Enterobacteriaceae, CRE, liver transplantation, intestinal colonization, CT screening, anal swabs

Mei Zhong,1,* Zhenwei Zhai,2,* Xing Zhou,1 Jingxia Sun,1 Hui Chen,1 Wensheng Lu1 1Department of Endocrinology, The Peoples Hospital of Guangxi Zhuang Autonomous Region, Guangxi, People’s Republic of China; 2Department of Endocrinology, Tongde Hospital of Yuncheng, Changzhi Medical College, Shanxi, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wensheng Lu Tel +86 17634045555Email lwswxqz@163.comAim: Gitelman syndrome (GS) is an autosomal recessive disorder characterized by hypokalemic metabolic alkalosis. In this study, we investigated the clinical presentation and sequenced 26 exons of SLC12A3 gene in a patient with a clinical suspicion of GS.Methods: Clinical work-up including clinical examination, electrocardiography (ECG), chest X-ray, bone mineral density (BMD), and ultrasound examination was conducted and all exons of SLC12A3 gene were analyzed by whole-exome sequencing.Results: The patient showed hypokalemia, hypomagnesemia, and metabolic alkalosis and was found to have four novel homozygous missense mutations including one known mutation (c.1456 G>A in exon 12) and three novel mutations (c.366A > G in exon 2, c.791C > G in exon 6 and c.1027C > T in exon 8).Conclusion: Four mutation sites of SLC12A3 gene were found in the patient, three of which have not been reported before. These results may be useful for better understanding the function of this gene and can assist clinicians with treatment decision-making.Keywords: Gitelman syndrome, clinical characteristics, SLC12A3 gene, exome sequencing, gene mutation

Xi Lin,1,2 Junwan Lu,1,2 Changrui Qian,1,2 Hailong Lin,1– 3 Qiaoling Li,1– 3 Xueya Zhang,1– 3 Hongmao Liu,1– 3 Zhewei Sun,1,2 Danying Zhou,1,2 Wei Lu,1,2 Mei Zhu,4 Hailin Zhang,2,3 Teng Xu,5 Kewei Li,1,2 Qiyu Bao,1– 3 Li Lin2,3 1Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China; 2Institute of Biomedical Informatics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China; 3The Second Affiliated Hospital and Children’s Hospital, Wenzhou Medical University, Wenzhou, 325027, People’s Republic of China; 4Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, Zhejiang, 310013, People’s Republic of China; 5Institute of Translational Medicine, Baotou Central Hospital, Baotou, 014040, People’s Republic of ChinaCorrespondence: : Li LinThe Second Affiliated Hospital and Yuying Children’s Hospital, Wenzhou Medical University, Wenzhou, 325027, People’s Republic of ChinaTel +86-577-88002134Email linli97@126.comQiyu BaoInstitute of Biomedical Informatics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of ChinaTel/Fax +86-577-86699398Email baoqy@genomics.cnPurpose: An increasing frequency of antibiotic resistance has been observed in both clinical and environmental Aeromonas hydrophila isolates in recent years. However, there are still very few in-depth studies regarding the role of plasmids in the antibiotic resistance of A. hydrophila. Hence, we investigated the molecular and functional characterization of a multidrug-resistant plasmid encoding an NDM-like metallo-β-lactamase, AFM-1, in the clinical A. hydrophila isolate SS332.Methods: The minimum inhibitory concentrations (MICs) of 24 antibiotics against A. hydrophila SS332 were measured by the agar dilution method. The genome of A. hydrophila SS332 was sequenced with PacBio and Illumina platforms. Six plasmid-borne antimicrobial resistance genes were chosen for cloning, including blaAFM-1, blaOXA-1, msr(E), mph(E), aac(6ʹ)-Ib10, and aph(3ʹ)-Ia. Phylogenetic analysis, amino acid sequence alignment, and comparative genomic analysis were performed to elucidate the active site requirements and genetic context of the blaAFM-1 gene.Results: A. hydrophila SS332 showed high levels of resistance to 15 antibiotics, especially those with MIC levels at or above 1024 μg/mL, including ampicillin, cefazolin, ceftriaxone, aztreonam, spectinomycin, and roxithromycin. Six plasmid-borne resistance genes from A. hydrophila were verified to be functional in E. coli DH5α. AFM-1 shared 86% amino acid identity with NDM-1 and showed resistance to ampicillin, cefazolin, cefoxitin, and ceftazidime. In addition, the blaAFM-1 gene was associated with three different novel ISCR19-like elements, designated ISCR19-1, ISCR19-2 and ∆ISCR19-3, which may be involved in the acquisition and mobilization of the blaAFM-1 gene.Conclusion: Our investigation showed that plasmid-borne resistance genes can contribute to antibiotic resistance in A. hydrophila SS332. A novel blaNDM-like gene, blaAFM-1, was verified to be functional and associated with novel ISCR19-like elements. This fact indicated the risk of spread of blaAFM-1 genes and ISCR19-like elements.Keywords: Aeromonas hydrophila, whole-genome sequencing, plasmid-borne resistance genes, blaAFM-1, ISCR19-like elements

Leixi Xue,1,* Yi Zhang,1,* Jiajun Xu,1 Wentian Lu,1 Qing Wang,1 Jinxiang Fu,2 Zhichun Liu1 1Department of Rheumatology and Immunology, The Second Affiliated Hospital of Soochow University, Suzhou, People’s Republic of China; 2Department of Hematology, The Second Affiliated Hospital of Soochow University, Suzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhichun LiuDepartment of Rheumatology and Immunology, The Second Affiliated Hospital of Soochow University, Sanxiang Road No. 1055, Suzhou, Jiangsu, 215004, People’s Republic of ChinaTel +86 13771994276Email liuzhichun5190@163.comJinxiang FuDepartment of Hematology, The Second Affiliated Hospital of Soochow University, Sanxiang Road No. 1055, Suzhou, Jiangsu, 215004, People’s Republic of ChinaTel +86 13962525217Email fjx3000@126.comPurpose: Current studies on the mechanism of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) in lupus nephritis (LN) mainly focus on the inflammatory pathway. Herein, we aimed to determine whether TWEAK could promote the progression of renal interstitial fibrosis by regulating peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) expression and intervening in lipid metabolism in LN.Materials and Methods: MRL/lpr mice, an animal model of lupus, were treated with the anti-TWEAK antibody or co-treated with adeno-associated virus-mediated PGC-1α short hairpin RNA (shRNA). In addition, human proximal tubular epithelial cells (HK2 cells) were treated with recombinant human TWEAK (rhTWEAK) or ammonium pyrrolidine dithiocarbamate (PDTC) in vitro.Results: The renal contents of free fatty acids and triglycerides were higher in MRL/lpr mice than in MRL/MpJ mice; however, these contents were decreased by treatment with the anti-TWEAK antibody. Based on immunofluorescence staining, the expression of PGC-1α was markedly more in the renal tubules of MRL/MpJ mice than in the glomeruli. However, treatment with anti-TWEAK antibody increased the levels of PGC-1α and its downstream target genes, which were remarkably lower in MRL/lpr mice than in MRL/MpJ mice. Anti-TWEAK antibody effectively eased renal interstitial fibrosis, which manifested as a decrease in the deposition of collagen fibers and the inhibition of type I collagen and fibronectin expression. However, the therapeutic effects of the anti-TWEAK antibody were abolished by PGC-1α shRNA. Treatment with rhTWEAK decreased PGC-1α expression in both dose- and time-dependent manners in HK2 cells in vitro. PDTC, an inhibitor of IκBα phosphorylation, suppressed the decrease in the PGC-1α protein level induced by rhTWEAK treatment.Conclusion: Our results suggest that TWEAK prevents renal tubular PGC-1α expression by promoting NF-κB activation, resulting in a deficiency in lipid metabolism and the progress of renal interstitial fibrosis. The upregulation of renal tubular PGC-1α expression to improve lipid metabolism is one of the mechanisms employed by the anti-TWEAK antibody to treat renal interstitial fibrosis.Keywords: TWEAK, lupus nephritis, renal interstitial fibrosis, PGC-1α, lipid metabolism

Yuejie Xu,1,* Lei Xu,1,* Weijing Zhao,1 Qing Li,1 Ming Li,1 Wei Lu,1 Hui Zeng,1 Jinhua Yan,2 Daizhi Yang,2 Wei Wu,3 Jianping Weng,2 Jiemin Pan,1 Fang Liu1 1Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai Clinical Center for Diabetes, Shanghai Key Clinical Center for Metabolic Disease, Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai, 200233, People’s Republic of China; 2Department of Endocrinology and Metabolic Disease, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, People’s Republic of China; 3Department of Pediatrics, Tongji Hospital, Wuhan Tongji Medical University, Wuhan, Hubei, 430030, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jiemin Pan; Fang LiuDepartment of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai Clinical Center for Diabetes, Shanghai Key Clinical Center for Metabolic Disease, Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, 600 Yishan Road, Xuhui District, Shanghai, 200233, People’s Republic of ChinaTel +86-18930173803Email lilypjm@hotmail.com; f-liu@sjtu.edu.cnPurpose: Smartphones have received increasing attention and achieved positive outcomes in diabetes intervention. The widespread use of WeChat in China provides an opportunity for self-management practices in patients with diabetes. Nevertheless, how to combine the strengths of the WeChat platform with traditional medical strategy remains to be explored. This study aimed to evaluate the efficacy of a novel flash glucose monitoring device combined with the WeChat platform in juvenile type 1 diabetes management.Patients and Methods: A total of 60 juvenile patients with type 1 diabetes were randomly assigned into three groups: a blood glucose self-monitoring group (group A), a flash glucose monitoring (group B), and a flash glucose monitoring combined WeChat-interactive management group (group C). The intergroup differences in demographics, biochemical indicators, and questionnaire scores of the Diabetes Monitoring and Treatment Satisfaction Questionnaire and Diabetes Specific Quality of Life assessment were compared at the baseline and after 6 months.Results: After the 6-month intervention, groups B and C showed significantly lower glycated hemoglobin A1c (HbA1c) levels compared to those observed at baseline (both P < 0.05), with the largest decrease observed in group C (group B vs group C, P =0.04). Hypoglycemic episodes per month decreased from baseline in groups B and C (both P < 0.05) and were more significant in group C (P < 0.001). In addition, the DMTSQ scores increased in the 6th month in all groups (all P < 0.05), and the largest rise in scores was found in group C, followed by groups B and A. The DQOL scores in groups B and C decreased significantly from the baseline (both P < 0.05), with no change in group A.Conclusion: Flash glucose monitoring combined with the WeChat-interactive system may help achieve sustained glycemic control and higher satisfaction in patients with juvenile type 1 diabetes.Trial Registration: This study was registered at chictr.org.cn, number ChiCTR1900025495. Registered 29 August 2019.Keywords: WeChat, type 1 diabetes, self-management, glycated hemoglobin A1c

Yunzhu Zhang,1 Wenxia Lu2 1Department of Obstetrics and Gynecology, Shanghai Pudong New District People’s Hospital, Shanghai, People’s Republic of China; 2Department of Medicine, Shanghai Tianyou Hospital, Shanghai, People’s Republic of ChinaCorrespondence: Wenxia Lu, Department of Medicine, Shanghai Tianyou Hospital, 528 Zhennan Road, Putuo District, Shanghai, People’s Republic of China, 200331, Tel/Fax +86- 21-66699900, Email drluwenxia@outlook.com

Ying Chu, Yi Chang, Wei Lu, Xiumei Sheng, Shengjun Wang, Huaxi Xu, Jie Ma Department of Immunology, Jiangsu Key Laboratory of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang 212013, People’s Republic of ChinaCorrespondence: Jie MaDepartment of Immunology, Jiangsu Key Laboratory of Laboratory Medicine, School of Medicine, Jiangsu University, 301 XueFu Road, Zhenjiang, Jiangsu Province 212013, People’s Republic of ChinaTel +86-1377-555-5586Email jsdxmajie@163.comAbstract: Autophagy is a critical cellular process that generally protects cells and organisms from harsh environment, including limitations in adenosine triphosphate (ATP) availability or a lack of essential nutrients. Metabolic reprogramming, a hallmark of cancer, has recently gained interest in the area of cancer therapy. It is well known that cancer cells prefer to utilize glycolysis rather than oxidative phosphorylation (OXPHOS) as their major energy source to rapidly generate ATP even in aerobic environment called the Warburg effect. Both autophagy and glycolysis play essential roles in pathological processes of cancer. A mechanism of metabolic changes to drive tumor progression is its ability to regulate autophagy. This review will elucidate the role and the mechanism of glycolysis in regulating autophagy during tumor growth. Indeed, understanding how glycolysis can modulate cellular autophagy will enable more effective combinatorial therapeutic strategies.Keywords: glycolysis, autophagy, lung cancer, lactate, metabolism

Qiuke Xiao,* Jinwei Ran,* Weizhong Lu, Ruijie Wan, Lujue Dong, Zhenyu Dai Department of Orthopedics, Chongqing Traditional Chinese Medicine Hospital, No.4 Clinical Medicine School of Chengdu University of Traditional Chinese Medicine, Chongqing 400021, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhenyu DaiDepartment of Orthopedics, Chongqing Traditional Chinese Medicine Hospital, No.4 Clinical Medicine School of Chengdu University of Traditional Chinese Medicine, Jiangbei District, Chongqing 400021, People’s Republic of ChinaTel/Fax +86 23 6798 3672Email zhenyudai@cdutcm.edu.cnBackground: Increasing attention has been paid to posttraumatic affective disorders. However, orthopedic surgeons dealing with trauma often ignore the harm of such diseases.Objective: To investigate the point prevalence and influencing factors of acute stress disorder (ASD) in elderly patients with osteoporotic fractures (EPOFs) from the perspective of orthopedic surgeons.Patients and Methods: A total of 595 cases of EPOFs were treated at our hospital from January 1, 2018, to June 30, 2019. The patients meeting our inclusion criteria were assessed using a structured interview based on the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) criteria to verify the presence of ASD. After diagnosis, the participants were divided into two groups (those with and without ASD). The sociodemographic characteristics, disease characteristics, and Social Support Rating Scale (SSRS) scores were assessed. The chi-square test was used for univariate analysis, and multivariate analysis was performed using binary logistic regression.Results: Of the 524 participants, 32 (6.1%) met the criteria for the diagnosis of ASD. The results of the univariate analysis showed that gender, personality, living alone, monthly family income, initial fear, poor prognosis expectation, anxiety/depression, pain, and social support were associated with ASD in EPOFs (P< 0.05). The multivariate regression analysis showed that isolation, low monthly family income, introversion, poor prognosis expectation, previous traumatic history, and intense pain were the main influencing factors and risk factors (OR> 1) for ASD in EPOFs.Conclusion: Being female, living alone, introversion, poor family income, intense initial fear, poor prognosis expectation, anxiety/depression, intense pain perception and low social support were significantly related to the occurrence of ASD in EPOFs. To achieve optimal recovery in EPOFs, orthopedic surgeons should meet both the physiological and psychological needs of the patients.Keywords: acute stress disorder, gender, trauma, fracture, elderly

Wenjie Lu,1,* Rukun Zang,1,* Yuanna Du,1 Xinghua Li,1 Hongwei Li,1 Chuan Liu,2 Yipeng Song,1 Yuncheng Li,3,* Yang Wang1 1Department of Radiation Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, Yantai, Shandong, People’s Republic of China; 2Department of Otorhinolaryngology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, People’s Republic of China; 3Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yipeng Song; Yang WangDepartment of Radiation Oncology, Yantai Yuhuangding Hospital, Affiliated Hospital of Qingdao University, 20 Yuhuangdi East Road, Yantai, Shandong 264000, People’s Republic of ChinaTel +8613361328765; +8615318695096Email syp1972@sina.com; wangyang2003@126.comBackground: Mitochondrial fission regulator 2 (MTFR2) has been reported to promote proliferation, migration and invasion in tumors; however, little is known about its function in breast cancer. Thus, we investigated the effect of MTFR2 expression on prognosis of breast cancer.Methods: The expression of MTFR2 in breast cancer tissues was detected by immunohistochemistry, and overall survival (OS) and recurrence free survival (RFS) were evaluated by the Log rank test and Cox model.Results: We found that MTFR2 expression was significantly associated with clinical stage (P< 0.001), T classification (P=0.005), N classification (P=0.001), M classification (P=0.041), HER2 expression (P= 0.001), and molecular subtypes (P=0.002), respectively. Compared with low MTFR2 expression, the patients with higher expression of MTFR2 exhibited significantly shorter OS and RFS (All P < 0.001). Both univariate and multivariate analyses showed that MTFR2 was an independent prognostic factor for OS (HR, 2.8, 95% CI 1.1– 6.8, P = 0.023) and RFS (HR, 2.8, 95% CI 1.2– 6.4, P = 0.015) in breast cancer patients. Moreover, in HER2 positive and TNBC subtype, the associations between high MTFR2 expression and poor OS and RFS were more pronounced.Conclusion: Taken together, our results demonstrated that high MTFR2 expression was associated with poor prognosis of breast cancer patients, and such an association was more pronounced in the patients with aggressive tumors. Therefore, MTFR2 expression might be a potentially important prognostic biomarker and clinical target for patients with breast cancer.Keywords: MTFR2, breast neoplasms, prognosis, biomarker, survival analysis

Guoqing Ouyang,1 Guangdong Pan,1 Yongrong Wu,1 Qiang Liu,1 Wuchang Lu,1 Xiang Chen2 1Department of Hepatobiliary Surgery, Liuzhou People’s Hospital, Liuzhou, Guangxi, People’s Republic of China; 2Department of General Surgery, The Second Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of ChinaCorrespondence: Xiang ChenDepartment of General Surgery, The Second Xiangya Hospital, Central South University, Renmin Road 139, Changsha 410011, Hunan, People’s Republic of ChinaEmail chenxiangxt@csu.edu.cnPurpose: Gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP) were involved in the development and progression of cancers. This study aimed to evaluate the prognostic value of a preoperative GGT:ALP ratio (GAR) in hepatocellular carcinoma (HCC) patients with curative liver resection.Patients and Methods: A total of 380 HCC patients underwent curative liver resection before December 2017 and from January to December 2018 were included and stratified into training set and validation set, respectively. Prediction accuracy was evaluated by the area under the receiver operating characteristic curve (AUC). Factors determined to be significant for overall survival (OS) and tumor-free survival (TFS) by using Cox regression analysis. The Kaplan–Meier method and Log rank test were utilized for survival analysis.Results: The AUC of GAR was 0.70 (P < 0.001). An optimal cut-off value of 0.91 yielded a sensitivity of 78.1% and a specificity of 60.4% for GAR (P < 0.001), which stratified the HCC patients into high-risk (> 0.91) and low-risk (≤ 0.91) groups. Time-dependent ROC revealed that the AUCs for 1-, 3-, and 5-year OS predictions for GAR were 0.60, 0.69 and 0.62, respectively. In addition, GAR was identified as an independent risk factor for OS and TFS both in training and validation cohort by univariate and multivariate Cox regression analysis, as well as a good prognostic indicator for patients with Barcelona Clinic Liver Cancer stage C or without vascular invasion. Notably, the AUC of the GAR for survival was better than several potential prognostic indices (P < 0.05).Conclusion: We identified the GAR as a prognostic indicator in two independent cohorts of HCC patients with curative liver resection. The patients with decreased GAR score were significantly associated with better OS and TFS.Keywords: hepatocellular carcinoma, gamma-glutamyltransferase, alkaline phosphatase, overall survival, tumor-free survival, prognostic indicator

Liping Li, Dan Ye, Liang Liu, Xiaoju Li, Jun Liu, Shengtian Su, Wenjing Lu, Zhigao Yu Department of Oncology, Xiantao First People’s Hospital, Xiantao, Hubei, People’s Republic of ChinaCorrespondence: Zhigao YuDepartment of Oncology, Xiantao First People’s Hospital, No. 29, Middle Section of Mianzhou Avenue, Xiantao, Hubei 433000, People’s Republic of ChinaTel +86 13477410999Email zhongjiyi330108@126.comPurpose: Non-small cell lung cancer (NSCLC) is a typical epithelial lung cancer with high metastasis, incidence and mortality. In recent years, long noncoding RNA small nucleolar RNA host gene 7 (SNHG7) has been identified as significant regulator in different cancer types, including NSCLC. However, the underlying molecular mechanism of SNHG7 during NSCLC tumorigenesis and progression remains largely unclear.Methods: SNHG7 and miR-181a-5p expression in NSCLC tumors and cells were detected by qRT-PCR. Cell viability, migration, invasion and apoptosis were evaluated by CCK-8, transwell and flow cytometry assay, respectively. A549 and NCI-H1299 xenograft mice model was constructed by subcutaneously injecting cells stably transfected with sh-SNHG7 and sh-NC. The interaction between SNHG7 and miR-181a-5p was validated by luciferase reporter system, RIP and RNA pull down assay. Protein expression of cleaved caspase 3, proliferating cell nuclear antigen (PCNA), AKT, p-AKT, mammalian target of rapamycin (mTOR) and p-mTOR was analyzed by Western blot.Results: SNHG7 expression was up-regulated while miR-181a-5p expression was down-regulated in NSCLC tumors, especially those from patients at Phase III+IV, compared with normal tissues. However, SNHG7 depletion attenuated tumor growth in vitro and in vivo. Moreover, miR-181a-5p inhibitor abolished SNHG7 silencing induced inhibition on proliferation, migration and invasion in NSCLC. Subsequently, we found SNHG7 modulated cell progression by targeting miR-181a-5p and activating AKT/mTOR signaling pathway.Conclusion: SNHG7 accelerates proliferation, migration and invasion of NSCLC by suppressing miR-181a-5p through AKT/mTOR signaling pathway, thus presenting desirable biomarkers for NSCLC therapy.Keywords: NSCLC, SNHG7, miR-181a-5p, AKT/mTOR pathway, progression

Kelie Chen,1,2,* Yuequn Niu,3,* Shengchao Wang,1,* Zhiqin Fu,4 Hui Lin,1,2 Jiaoying Lu,2 Xinyi Meng,2 Bowen Yang,2 Honghe Zhang,1 Yihua Wu,1,2 Dajing Xia,1,2 Weiguo Lu1 1Department of Gynecologic Oncology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310058, People’s Republic of China; 2Department of Toxicology, School of Public Health, School of Medicine, Zhejiang University, Hangzhou 310058, People’s Republic of China; 3Department of Thoracic Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, People’s Republic of China; 4Department of Gynecological Oncology, Zhejiang Cancer Hospital, Hangzhou 310022, People’s Republic of China*These authors contributed equally to this workCorrespondence: Dajing XiaDepartment of Toxicology, School of Public Health, School of Medicine, Zhejiang University, 866 Yuhangtang Road, Hangzhou, Zhejiang 310058, People’s Republic of ChinaTel/ Fax +86-0571-88208140Email dxia@zju.edu.cnWeiguo LuDepartment of Gynecologic Oncology, Women’s Hospital, School of Medicine, Zhejiang University, 1 Xueshi Road, Hangzhou, Zhejiang 310006, People’s Republic of ChinaTel +86-571-8706-1501Fax +86-571-87061878Email lbwg@zju.edu.cnBackground: Heterogeneity plays an essential role in ovarian cancer. Patients with different clinical features may manifest diverse patterns in diagnosis, treatment, and prognosis. The aim of the present study was to identify a novel ovarian cancer–classification model through cluster analysis and assess its significance in prognosis.Methods: Among patients diagnosed with ovarian cancer in the Women’s Hospital School of Medicine, Zhejiang University between January 2014 and May 2019, 328 patients were included in a K-mean cluster analysis and 176 patients followed up. Major clinical indicators, overall survival, and recurrence-free survival in different subgroups were compared.Results: Two clusters for ovarian cancer were identified and grouped as noninflammatory (n=247) and inflammatory subtypes (n=81). Compared with the noninflammatory subgroup, the inflammatory subgroup presented a statistically significantly higher level of median CRP (median (IQR) 20.4 [7.8– 47.3] vs 1.2 [0.4– 3.5], p< 0.001), neutrophil percentage (median (IQR) 76.9 [72.6– 81.3] vs 66.2 [61.0– 72.0], p< 0.001), leukocyte count (median (IQR) 8.9 [7.0– 10.0] vs 6.0 [5.1– 7.2], p< 0.001), fibrinogen (median (IQR) 5.0 [4.4– 6.0] vs 3.4 [2.9– 3.9], p< 0.001), and platelet count (median (IQR) 324 [270– 405] vs 229 [181.5– 269], p< 0.001). During a median follow-up of 52 months, 21 participants (16.3%) died in the noninflammatory group, while 14 (29.8%) died in the inflammatory group (HR 2.15, 95% CI 1.09– 4.23; p=0.024). Death/recurrence was observed in 38 (29.5%) patients from the noninflammatory group and 25 (53.2%) from the inflammatory group (HR 2.32, 95% CI 1.40– 3.85; p< 0.001).Conclusion: Our study revealed a novel classification model of ovarian cancer that features inflammation. Inflammation predicts shorter survival and poorer prognosis, suggesting the significance of inflammation in the management of ovarian cancer.Keywords: ovarian cancer, classification, heterogeneity, cluster analysis, inflammation, prognosis

Zihang Feng,1,* Wenxia Lu,2,* Yu Xie3 1Southern Medical University, Guangzhou, Guangdong Province, People’s Republic of China; 2Nanjing Medical University, Nanjing, Jiangsu Province, People’s Republic of China; 3Obstetrics and Gynecology Hospital of Fudan University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu Xie, Obstetrics and Gynecology Hospital of Fudan University, 419 Fangxie Road, Shanghai 200011, People’s Republic of China, Tel +86-021-33189900, Fax +86-021-63450900, Email yuxie18@fudan.edu.cn View the original paper by Mr Ashrafizadeh and colleagues

Shizhou Yang,1,* Tingting Chen,1,* Lu Huang,2 Shanshan Xu,1 Zhu Cao,1 Songfa Zhang,1 Junfen Xu,1 Yang Li,1 Yongfang Yue,1 Weiguo Lu,1,3 Xiaodong Cheng,1,3 Xing Xie1,3 1Department of Gynecologic Oncology, Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People’s Republic of China; 2Zhejiang Provincial People’s Hospital, Hangzhou, Zhejiang, People’s Republic of China; 3Center for Uterine Cancer Diagnosis & Therapy Research of Zhejiang Province, Hangzhou, Zhejiang, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xing Xie Tel +86 571 8706 1501Fax +86 571 8706 1878Email xiex@zju.edu.cnPurpose: To determine whether early proteins from high-risk human papillomavirus (HPV) have the capacity to maintain cellular stemness.Patients and methods: First, we isolated cancer stem cell like cells from two cervical cancer cell lines, SiHa and CaSki, using non-adhesive culture with serum-free medium. Second, we knocked down HPV16 E7 in SiHa sphere cells and overexpressed HPV16 E7 in U2OS sphere cells. Third, we used RNA-seq analysis and Western blotting to screen and identify the expression of differentially expressed genes in SiHa cells with HPV16 E7 knockdown.Results: We found that both SiHa and CaSki cells grew as cell spheres (oncospheres) and shared the properties of cancer stem cells, including high expression of stem cell marker OCT4 and SOX2, self-renew, and resistance to chemotherapeutic drugs. The stem-like properties were deprived when HPV16 E7 was knocked down in SiHa sphere cells and maintained when HPV16 E7 was over-expressed in U2OS sphere cells. APH1B was up-regulated, among differential expression genes, in SiHa cells with HPV16 E7 knockdown and modulated cellular stemness and SiHa sphere cells with APH1B knockdown regained the stem-like properties deprived by E7 inhibition.Conclusion: HPV16 E7 possesses the capacity to maintain cellular stemness and APH1B may participate in this process in cervical cancer sphere cells.Keywords: cervical cancer, HPV, cancer stem cell, APH1B

Jin Li1,*, Weiwei Hu2,*, Min Li1, Shaoli Deng1, Qing Huang1, Weiping Lu1 1Department of Clinical Laboratory Medicine, Institute of Surgery Research, Daping Hospital, Army Medical University, Chongqing 400042, People’s Republic of China; 2Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weiping LuDepartment of Clinical Laboratory Medicine, Institute of Surgery Research, Daping Hospital, Army Medical University, Chongqing 400042, People’s Republic of ChinaTel +86 236 875 7607Fax +86 236 871 6530Email luweiping19710416@163.comBackground: Metallo-β-lactamase-producing Pseudomonas aeruginosa (MBL-PA) are important causative agents of nosocomial infections and are associated with significant mortality rates, especially in intensive care units. The timely detection and typing of these strains is essential for surveillance, outbreak prevention and antibiotic therapy optimization. In this study, fifteen VIM-type and fifteen SPM-type MBL-PA strains were selected as strains to establish MALDI-TOF MS SuperSpectra.Methods: This study was undertaken to evaluate the utility of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with the VITEK MS plus system in the detection of VIM- and SPM-type MBL-PA isolates. For each species, we increased the reference spectra, and then, a SuperSpectrum was created based on the selection of 39 specific masses. In a second step, we validated the SuperSpectra with the remaining 50 isolates (25 isolates of VIM-type and 25 isolates of SPM-type).Results: Fifty MBL-PA strains were used as the validation strains, including twenty-five VIM-type and twenty-five SPM-type MBL-PA strains. Complete antimicrobial susceptibility testing and genotypic characterizations were performed for all isolates, which were subsequently identified using the newly created SuperSpectra databases following a previously reported method. The results showed that there was 92% agreement between the MBL profile generated by MALDI-TOF MS and that obtained using gene sequencing analysis methods.Conclusion: MALDI-TOF MS is a promising, rapid and economical method for detecting VIM- or SPM-type MBL-PA that could be successfully introduced into the routine diagnostic workflow of clinical microbiology laboratories.Keywords: VIM, SPM, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, SuperSpectra, VITEK MS plus system

Mathew Suji Eapen,1 Pawan Sharma,1–3 Archana Vijay Gaikwad,1 Wenying Lu,1 Stephen Myers,1 Philip M Hansbro4,5, Sukhwinder Singh Sohal11Respiratory Translational Research Group, Department of Laboratory Medicine, College of Health and Medicine, University of Tasmania, Launceston, TAS 7248, Australia; 2Medical Sciences, University of Technology Sydney, Sydney, NSW 2007, Australia; 3Woolcock Emphysema Centre, Woolcock Institute of Medical Research, University of Sydney, Sydney, NSW 2037, Australia; 4Priority Research Centre for Healthy Lungs, Hunter Medical Research Institute and the University of Newcastle, Newcastle, NSW 2308, Australia; 5Centre for Inflammation, Centenary Institute and University of Technology Sydney, Sydney, NSW 2007, AustraliaAbstract: COPD is a common and highly destructive disease with huge impacts on people and health services throughout the world. It is mainly caused by cigarette smoking though environmental pollution is also significant. There are no current treatments that affect the overall course of COPD; current drugs focus on symptomatic relief and to some extent reducing exacerbation rates. There is an urgent need for in-depth studies of the fundamental pathogenic mechanisms that underpin COPD. This is vital, given the fact that nearly 40%–60% of the small airway and alveolar damage occurs in COPD well before the first measurable changes in lung function are detected. These individuals are also at a high risk of lung cancer. Current COPD research is mostly centered around late disease and/or innate immune activation within the airway lumen, but the actual damage to the airway wall has early onset. COPD is the end result of complex mechanisms, possibly triggered through initial epithelial activation. To change the disease trajectory, it is crucial to understand the mechanisms in the epithelium that are switched on early in smokers. One such mechanism we believe is the process of epithelial to mesenchymal transition. This article highlights the importance of this profound epithelial cell plasticity in COPD and also its regulation. We consider that understanding early changes in COPD will open new windows for therapy.Keywords: epithelial-to-mesenchymal transition, EMT, cancer, fibrosis, inflammation, HuR, EGFR, MMP, TGFβ

Lijun Xue,1,2 Yang Yizhi Chen,1 Zhiyun Yan,3 Wei Lu,1 Dong Wan,4 Huifeng Zhu11College of Pharmaceutical Sciences and Traditional Chinese Medicine, Southwest University, Chongqing 400715, People’s Republic of China; 2College of Pharmaceutical Engineering, Chongqing Chemical Industry Vocational College, Chongqing, 400020, People’s Republic of China; 3Wuhan Wusteel Good Life Service Co. LTD, Wuhan, 430000, People’s Republic of China; 4Department of Emergency and Critical Care Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of ChinaAbstract: Staphylococcus aureus is an important and common Gram-positive bacteria which causes clinical infections and food-poisoning cases. Therapeutic schedules for treatment of S. aureus infections are facing a challenge because of the emergence of multidrug resistance strains. It is urgent to find new antiinfective drugs to control S. aureus infection. S. aureus strains are capable of producing the golden carotenoid pigment: staphyloxanthin, which acts as an important virulence factor and a potential target for antivirulence drug design. This review is aimed at presenting an updated overview of this golden carotenoid pigment of S. aureus from the biosynthesis of staphyloxanthin, its function, and the genes involved in pigment production to staphyloxanthin: a novel target for antivirulence therapy.Keywords: staphyloxanthin, target, antiinfective drug, antivirulence therapy

Qiao Liu,1,2,* Zhongqi Li,1,* Ye Ji,1 Leonardo Martinez,3 Ui Haq Zia,4 Arshad Javaid,4 Wei Lu,2 Jianming Wang1,51Department of Epidemiology, Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, People’s Republic of China; 2Department of Chronic Communicable Disease, Center for Disease Control and Prevention of Jiangsu Province, Nanjing, People’s Republic of China; 3Division of Infectious Diseases and Geographic Medicine, School of Medicine, Stanford University, Stanford, CA, USA; 4Faculty of Public Health and Social Sciences, Khyber Medical University, Peshawar, Pakistan; 5Key Laboratory of Infectious Diseases, School of Public Health, Nanjing Medical University, Nanjing, People’s Republic of China*These authors contributed equally to this workObjective: Forecasting the seasonality and trend of pulmonary tuberculosis is important for the rational allocation of health resources; however, this foresting is often hampered by inappropriate prediction methods. In this study, we performed validation research by comparing the accuracy of the autoregressive integrated moving average (ARIMA) model and the back-propagation neural network (BPNN) model in a southeastern province of China.Methods: We applied the data from 462,214 notified pulmonary tuberculosis cases registered from January 2005 to December 2015 in Jiangsu Province to modulate and construct the ARIMA and BPNN models. Cases registered in 2016 were used to assess the prediction accuracy of the models. The root mean square error (RMSE), mean absolute percentage error (MAPE), mean absolute error (MAE) and mean error rate (MER) were used to evaluate the model fitting and forecasting effect.Results: During 2005–2015, the annual pulmonary tuberculosis notification rate in Jiangsu Province was 56.35/100,000, ranging from 40.85/100,000 to 79.36/100,000. Through screening and comparison, the ARIMA (0, 1, 2) (0, 1, 1)12 and BPNN (3-9-1) were defined as the optimal fitting models. In the fitting dataset, the RMSE, MAPE, MAE and MER were 0.3901, 6.0498, 0.2740 and 0.0608, respectively, for the ARIMA (0, 1, 2) (0, 1, 1)12 model, 0.3236, 6.0113, 0.2508 and 0.0587, respectively, for the BPNN model. In the forecasting dataset, the RMSE, MAPE, MAE and MER were 0.1758, 4.6041, 0.1368 and 0.0444, respectively, for the ARIMA (0, 1, 2) (0, 1, 1)12 model, and 0.1382, 3.2172, 0.1018 and 0.0330, respectively, for the BPNN model.Conclusion: Both the ARIMA and BPNN models can be used to predict the seasonality and trend of pulmonary tuberculosis in the Chinese population, but the BPNN model shows better performance. Applying statistical techniques by considering local characteristics may enable more accurate mathematical modeling.Keywords: ARIMA, BPNN, tuberculosis, incidence, forecasting

Wenxue Mao,1 Fan Wu,2 Robert J Lee,3 Weigen Lu,4 Jianxin Wang11Department of Pharmaceutics, School of Pharmacy, Fudan University & Key Laboratory of Smart Drug Delivery, Ministry of Education, Shanghai 201203, People’s Republic of China; 2College of Medicine, Des Moines University, Des Moines, IA 50312, USA; 3Division of Pharmaceutics and Pharmaceutical Chemistry, Ohio State University, Columbus, OH 43210, USA; 4China State Institute of Pharmaceutical Industry, Shanghai 201203, People’s Republic of ChinaBackground: Vincristine is a potent therapeutic agent with well-defined activity against hematologic malignancies and solid tumors. It is a cell-cycle specific drug with concentration and exposure duration dependent activity. When used by liposomal delivery, it exhibits enhanced anti-tumor activity. However, vincristine liposome formulation in the clinic is supplied as a 3-vial-kit due to lacking sufficient stability. So it has to be prepared in situ prior to use through a multi-step process.Purpose: The purpose here is to develop a more stable and ready-to-use liposomal formulation for vincritstine in one vial.Patients and methods: A series of preparations were investigated based on sphingomyelin/cholesterol/PEG2000-DSPE lipid composition, with different drug/lipid (D/L) ratios (1/10, 1/5, 1/2), using an active sucrose octasulfate triethylamine salt gradient loading method. In this work, compared to generic vincristine sulfate liposome injection (GVM), the stability both in vivo and in vitro and efficacy in vivo of novel vincristine liposomes were investigated.Results: It was shown that the degradation of vincristine during 2–8°C storage was significantly decreased from 8.2% in 1 month (GVM) to 2.9% in 12 months (D/L ratio 1/5). The half-time for sphingomyelin/cholesterol/PEG2000-DSPE liposomes in vivo could be adjusted from 17.4 h (D/L ratio 1/10) to 22.7 h (D/L ratio 1/2) in rats, while the half-time for GVM was only 11.1 h. The increase in drug retention contributed to the lower in vivo toxicity. The antitumor efficacy was evaluated using a human melanoma tumor model and showed remarkable improvement compared to GVM.Conclusion: The study demonstrates that the new formulation with the drug/lipid ratio of 1/5 owns a higher encapsulation efficiency, better stability, lower toxicity and superior antitumor efficacy, which is screened out for further development.Keywords: vincristine sulfate, liposome, TEA-SOS, storage stability, and anti-tumor efficacy

Yini Ma,1,2 Liying Guan,1 Yanxin Han,1 Yi Zhou,1 Xiaoming Li,1 Yumei Liu,1 Xiujuan Zhang,1 Weiying Zhang,1 Xiaohong Li,1 Shuhua Wang,1 Weidong Lu11Health Management Center, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, 250021, People’s Republic of China; 2Department of Nephrology, The Affiliated Hospital of Shandong Academy of Medical Sciences, Jinan 250031, People’s Republic of China Purpose: We have previously reported that PRDX2 plays an oncogenic role in colon cancer. In this study, the mRNA expression profile of PRDX2 in HCT116 cells was investigated. Furthermore, we selected Dynamin 3 (DNM3), which is up-regulated by siPRDX2, to investigate its expression pattern and functions in colon cancer.Patients and methods: PRDX2 siRNA was transfected into HCT116 cells and the mRNA profile was tested by RNA-Sequencing. The expression of interest proteins was determined by Western blot. DNM3 expression in colon cancer tissues and para-carcinoma tissues was evaluated by Western blot and immunohistochemistry assays. Full-length cDNA of DNM3 was cloned into pcDNA3.1 and introduced into HCT116 and HT29 cells. Cell proliferation was tested by CCK-8 and colony formation assays. Cell invasion and migration were tested by transwell assays. Gelatin zymography was utilized for detection of MMP9 activity. Cell apoptosis was investigated with Annexin V/PI staining and flow cytometry and visualized with Hoechst/PI staining assay. All statistical analysis was performed with SPSS 17.0 software.Results: PRDX2 knockdown led to 210 up-regulated genes and 16 down-regulated genes in HCT116 cells. We also found that DNM3 expression was up-regulated following PRDX2 silencing in HCT116 and HT29 cells. In colon cancer patients, DNM3 was down-regulated and showed a significant association with pathologic grading. DNM3 overexpression inhibited cell proliferation and induced apoptosis in HCT116 and HT29 cells. Cell migration and invasion were also down-regulated in DNM3 overexpressing colon cancer cells, which might be due to the inhibition of MMP9 proteolytic activities. After thorough investigation of the potential mechanism involved, we hypothesized that DNM3 overexpression induced activation of the mitochondrial apoptosis pathway and inhibition of the AKT pathway.Conclusion: These data suggest that DNM3 is down-regulated in colon cancer, serving as a tumor suppressor. Our study provides new sights into the prognostic value and therapeutic application of DNM3 in colon cancer.Keywords: mRNA expression profile, proliferation, invasion; migration, apoptosis, AKT

Jian Qiu,1,2 Yunfeng Fu,1 Jiewei Xu,3 Xiaohong Huang,2 Guorong Yao,2 Weiguo Lu1 1Department of Gynecologic Oncology, Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, China; 2Department of Obstetrics and Gynaecology, Huzhou Central Hospital, Huzhou, Zhejiang 313000, China; 3Department of General Surgery, Huzhou Central Hospital, Huzhou, Zhejiang 313000, China Purpose: The purpose of this study was to investigate the efficiency, complications, and subsequent fertility outcomes of two methods in treating patients with cesarean scar pregnancy (CSP) after receiving uterine artery embolization (UAE) treatment.Patients and methods: A total of 62 CSP patients who visited our hospital and underwent UAE from January 2013 to January 2018 were retrospectively analyzed. Patients were either treated by dilation and curettage (D&C) guided by ultrasonography or hysteroscopy. The differences of related clinical indicators, clinical efficacy, complications, and subsequent fertility outcomes between the two groups were analyzed.Results: The rates of therapeutic success of the ultrasonography group and hysteroscopy group were 84.6% and 95.7%, respectively, the difference was not statistically significant (P=0.243). However, the intraoperative blood loss, duration of hospitalization, and overall complications were significantly lower in hysteroscopy group compared with D&C guided by ultrasonography group (P

Wenbin Lu,1,* JingJing Ji,1,* Genshan Ma,1 Qiming Dai,1 Lijuan Chen,1 Pengfei Zuo,1 Yuanjin Zhao2 1Department of Cardiology, ZhongDa Hospital Affiliated with Southeast University, Nanjing, China; 2State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China *These authors contributed equally to this work Background: The two-dimensional incubation method is now the most commonly method for mesenchymal stem cell (MSC) production. however, gene expression and secretion of growth factors are relatively low; thus, the transplanted cells cannot be effectively utilized for potential clinical applications after acute myocardial infarction (AMI).Objectives: We aimed to investigate whether our newly made substrates of inverse opal with specific surface microstructures for MSC culturing can increase the viability of the cells and can contributes to decreased myocardial remodeling after transplanted to AMI mice. Methods: The inverse opal structure is fabricated by the convenient bottom-up approach of the self-assembly of colloidal nanoparticles. Mouse-derived MSCs were then cultured on the substrates when expanded at different times to investigate the cell growth status including morphology. Then the inverse opal substrates loaded MSCs were transplanted to AMI mice, cardiomyocyte apoptosis and LV remodeling were further compared. To explore the possible mechanisms of curation, the secretions and viability of MSCs on substrates were determined using mice ELISA kits and JC-1 mitochondrial membrane potential assay kits respectively at normal and hypoxic conditions. Results: 6 times expanded inverse opals allowed greatly the orderly growth of MSCs as compared to four (34% ± 10.6%) and two (20%±7.2%) times expanded as well as unexpanded (13%±4.1%) (P

Hao Cheng, Yong Zhang, Weiping Lu, Xianzhong Gao, Chenjie Xu, Hongguang Bao Department of Anesthesiology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China Background: Management of neuropathic pain is still a clinical challenge. Evidence has accumulated indicating that propolis is effective in attenuating neuropathic pain; however, the mechanism is not fully understood. Our present study investigated the effects and the possible mechanism of caffeic acid phenethyl ester (CAPE), the main ingredient of propolis, in improving neuropathic pain via its inhibition on p38/NF-κB signal pathway in microglia. Materials and methods: Chronic constriction injury (CCI) mice model and the microglial cell line BV-2 were used to investigate the effects and the mechanism of CAPE. Cell signaling was measured by real-time PCR, Western blotting and immunofluorescence assay. Results: CAPE relieved neuropathic pain behaviors induced by CCI in mice. CAPE also inhibited CCI-induced activation of microglia. Furthermore, CAPE suppressed the phosphorylation of p38 mitogen-activated protein kinase, inhibited the translocation of NF-κB and decreased the expression of proinflammatory cytokines tumor necrosis factor-α, IL-1β and IL-6. Conclusion: CAPE was found to be an effective and safe drug candidate for alleviating neuropathic pain by its powerful inhibition on the P38/NF-κB signal pathway. Keywords: caffeic acid phenethyl ester, NF-κB, neuropathic pain, microglia

Zhenyu Dai,1,* Yue Li,2,* Yonggang Yan,3 Ruijie Wan,1 Qiang Ran,1 Weizhong Lu,1 Bo Qiao,4 Hong Li3 1Department of Orthopedics, Chongqing Traditional Chinese Medicine Hospital, Chongqing, People’s Republic of China; 2Department of Clinical Laboratory, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, People’s Republic of China; 3College of Physical Science and Technology, Sichuan University, Chengdu, People’s Republic of China; 4Department of Orthopedics, the First Affiliated Hospital, Chongqing Medical University, Chongqing, People’s Republic of China *These authors contributed equally to this work Objective: To evaluate the internal fixation effect of nano-calcium-deficient hydroxyapatite/poly-amino acid (n-CDHA/PAA) composite screws in the intraarticular fracture model. Materials and methods: A total of 35 New Zealand White rabbits were used in a bilateral femoral intercondylar fracture model and randomly divided into two groups. n-CDHA/PAA screws were used in the experimental group, and medical metal screws were used in the control group. The fracture condition, range of motion, and the screw push-out strength were assessed, and an arthroscopic examination of knee joint was performed at 4, 8, and 12 weeks after surgery. The biodegradation of the n-CDHA/PAA screws in vivo was tested through weighing, and changes in screw structure were assessed by X-ray diffraction at 12 weeks after surgery. Results: The general situation of all animals was good and showed no incision infection and dehiscence after surgery. X-ray scanning showed that significant callus growth was present in both groups at 4 weeks after surgery, and there was no significant difference (P>0.05) in the Lane-Sandhu score between the experimental and control groups at all time points after surgery. There were no statistically significant differences (P>0.05) in the range of motion and Oswestry Arthroscopy Score of arthroscopic examination of the knee joints between the two groups. The screw push-out strength of the control group was stronger than that of the experimental group at 4 weeks after surgery (P0.05). The degradation tests showed that the n-CDHA/PAA screws degraded gradually after implantation, and the weight loss rate was approximately 16% at 12 weeks after surgery. The X-ray diffraction results showed that the crystal structure of the outer surface of the n-CDHA/PAA screw has changed at 12 weeks after surgery. Conclusion: The n-CDHA/PAA screw is an effective and safe implant as a potential internal fixation device for an intercondylar fracture of the femur, and its internal fixation effect was similar to that of medical metal screw. Keywords: n-CDHA/PAA screw, internal fixation, intraarticular fracture, biocompatibility, bioactivity