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Chang-Sheng Ma,1,2,* Ya-Ping Ma,1,3,* Bo Han,1,2,* Wan-Li Duan,1 Shu-Chen Meng,1 Min Bai,1 Hao Dong,1 Li-Ying Zhang,1 Meng-Yuan Duan,1,2 Jing Liu,1 Ai-Jun Deng,2 Mao-Tao He1,2 1Department of Basic Medical Sciences, Shandong Second Medical University, Weifang, Shandong, People’s Republic of China; 2Department of Ophthalmology, Affiliated Hospital of Shandong Second Medical University, Weifang, People’s Republic of China; 3Department of Pathology, The 942Hospital of the People’s Liberation Army Joint Logistic Support Force, Yinchuan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Mao-Tao He, Department of Basic Medical Sciences, Shandong Second Medical University, Weifang, Shandong, People’s Republic of China, Tel +86-18209617186, Fax +86-0536-3081201, Email hemaotao@sdsmu.edu.cn Ai-Jun Deng, Department of Ophthalmology, Affiliated Hospital of Shandong Second Medical University, Weifang, People’s Republic of China, Email dengaijun@hotmail.comPurpose: Ischemic stroke is a refractory disease wherein the reperfusion injury caused by sudden restoration of blood supply is the main cause of increased mortality and disability. However, current therapeutic strategies for the inflammatory response induced by cerebral ischemia-reperfusion (I/R) injury are unsatisfactory. This study aimed to develop a functional nanoparticle (MM/ANPs) comprising apelin-13 (APNs) encapsulated in macrophage membranes (MM) modified with distearoyl phosphatidylethanolamine-polyethylene glycol-RVG29 (DSPE-PEG-RVG29) to achieve targeted therapy against ischemic stroke.Methods: MM were extracted from RAW264.7. PLGA was dissolved in dichloromethane, while Apelin-13 was dissolved in water, and CY5.5 was dissolved in dichloromethane. The precipitate was washed twice with ultrapure water and then resuspended in 10 mL to obtain an aqueous solution of PLGA nanoparticles. Subsequently, the cell membrane was evenly dispersed homogeneously and mixed with PLGA-COOH at a mass ratio of 1:1 for the hybrid ultrasound. DSPE-PEG-RVG29 was added and incubated for 1 h to obtain MM/ANPs.Results: In this study, we developed a functional nanoparticle delivery system (MM/ANPs) that utilizes macrophage membranes coated with DSPE-PEG-RVG29 peptide to efficiently deliver Apelin-13 to inflammatory areas using ischemic stroke therapy. MM/ANPs effectively cross the blood-brain barrier and selectively accumulate in ischemic and inflamed areas. In a mouse I/R injury model, these nanoparticles significantly improved neurological scores and reduced infarct volume. Apelin-13 is gradually released from the MM/ANPs, inhibiting NLRP3 inflammasome assembly by enhancing sirtuin 3 (SIRT3) activity, which suppresses the inflammatory response and pyroptosis. The positive regulation of SIRT3 further inhibits the NLRP3-mediated inflammation, showing the clinical potential of these nanoparticles for ischemic stroke treatment. The biocompatibility and safety of MM/ANPs were confirmed through in vitro cytotoxicity tests, blood-brain barrier permeability tests, biosafety evaluations, and blood compatibility studies.Conclusion: MM/ANPs offer a highly promising approach to achieve ischemic stroke-targeted therapy inhibiting NLRP3 inflammasome-mediated pyroptosis. Keywords: apelin-13, cerebral ischemia-reperfusion injury, macrophage membrane, ischemic stroke therapy, pyroptosis

Abstract The vertebrate stress response (SR) is mediated by the hypothalamic–pituitary–adrenal (HPA) axis and contributes to generating context appropriate physiological and behavioral changes. Although the HPA axis plays vital roles both in stressful and basal conditions, research has focused on the response under stress. To understand broader roles of the HPA axis in a changing environment, we characterized an adaptive behavior of larval zebrafish during ambient illumination changes. Genetic abrogation of glucocorticoid receptor (nr3c1) decreased basal locomotor activity in light and darkness. Some key HPI axis receptors (mc2r [ACTH receptor], nr3c1), but not nr3c2 (mineralocorticoid receptor), were required to adapt to light more efficiently but became dispensable when longer illumination was provided. Such light adaptation was more efficient in dimmer light. Our findings show that the HPI axis contributes to the SR, facilitating the phasic response and maintaining an adapted basal state, and that certain adaptations occur without HPI axis activity.

Feng Pan,1,* Ai-ting Lu,1,2,* Xiaowei Mao,1,* Fang Hu,1 Hai Zhang,1 Baohui Han1 1Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 2Department of Pulmonary and Critical Care Medicine, Regional Medical Center for National Institute of Respiratory Diseases, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Baohui Han, Email 18930858216@163.comObjective: To investigate the knowledge of pulmonary rehabilitation (PR) among physicians involved in pulmonary disease management.Methods: This multi-regional cross-sectional survey was conducted from December 12, 2019 to January 22, 2020. The participants were enrolled and an electronic questionnaire was exclusively sent to the members of the Lung Cancer Special Committee of the China Medicine Education Association through the WeChat platform. Multivariable logistic regression analysis was performed to explore the associated factors of high PR knowledge scores (≥ 18 points).Results: From the 858 valid questionnaires, the routine implementation of PR was only reported for 16.95% of physicians. The main reason hindering the implementation of PR for patients was the limited knowledge and awareness of PR among the physicians involved (69.1%). A total of 618 and 240 physicians had high and low knowledge scores, respectively. Multivariable analysis suggests that the self-perception of PR knowledge (OR = 1.89, 95% CI: 1.32– 2.771, P = 0.001) was independently associated with high knowledge scores, while having no theoretical knowledge of PR was associated with poor knowledge scores (OR = 0.43, 95% CI: 0.26– 0.72, P = 0.001).Conclusion: Inadequate knowledge of pulmonary rehabilitation is evident among physicians who are involved in pulmonary disease management in China. This underscores the need for more comprehensive and standardized training to bolster their awareness and effective utilization of pulmonary rehabilitation.Keywords: chronic obstructive pulmonary disease, pulmonary rehabilitation, lung cancer, knowledge, oncologists

Yuhan Teng,1 Yining Jian,2 Xinyue Chen,3 Yang Li,4 Bing Han,2 Lei Wang3 1Department of Clinical Medicine, Southwest Medical University, Luzhou, Sichuan, People’s Republic of China; 2Department of Public Health, China Medical University, Shenyang, Liaoning, People’s Republic of China; 3Department of General Practice, the First Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of China; 4Department of General Practice, Hunnan Zhujia Community Health Service Center, Shenyang, Liaoning, People’s Republic of ChinaCorrespondence: Lei Wang, Department of General Practice, The First Hospital of China Medical University, 122 N Nanjing St., Heping District, Shenyang, 110001, People’s Republic of China, Fax +86-24-83283333, Email wanglei_tz@163.comPurpose: To predict the future number of patients with chronic obstructive pulmonary disease (COPD) in China and compare the three prediction models.Methods: A generalized additive model (GAM), autoregressive integrated moving average (ARIMA) model, and curve-fitting method were used to fit and predict the number of patients with COPD in China. Data on the number of patients with COPD in China from 1990 to 2019 were obtained from the Global Burden of Disease (GBD) database. The coefficient of determination (R2), mean absolute error (MAE), mean absolute percentage error (MAPE), root mean squared error (RMSE), relative error of prediction, Akaike Information Criterion (AIC), and Bayesian Information Criterion (BIC) were used to evaluate and compare the fitting effect, prediction effect, and reliability of the three models.Results: The GAM, ARIMA, and curve-fitting methods could predict future trends in COPD in China. The performance of the GAM is the best among the three models, whereas the curve fitting method is the worst, and the ARIMA (0,1,2) model is in between. The prediction results of the three models showed that the number of patients with COPD in China is expected to increase from 2020 to 2025.Conclusion: GAM and AIRMA models are recommended for predicting the future prevalence of COPD in China. The number of patients with COPD in China is expected to increase in the next few years. The prevention and control of COPD in China still needs to be strengthened. Using appropriate models to predict future trends in COPD will provide support for health policymakers.Keywords: generalized additive model, ARIMA model, curve fitting method, COPD, prediction

Yue Cao, Yue-Yue Feng, Wei Du, Jing Li, Ya-Lan Fei, Hao Yang, Meng Wang, Shi-Jie Li, Xian-Jin Li, Bing Han Division of Cardiology, Xuzhou Central Hospital, Xuzhou, People’s Republic of ChinaCorrespondence: Bing Han, Division of Cardiology, Xuzhou Central Hospital, 199 South Jiefang Road, Xuzhou, 221009, People’s Republic of China, Tel +86-516-83985069, Fax +86-516-83956012, Email hbing777@hotmail.comPurpose: To investigate the reasons for elderly atrial fibrillation (AF) patients not continuing their oral anticoagulation (OAC) treatment and the factors that influence this behavior.Methods: Elderly AF patients (aged≥ 75 years) hospitalized from December 2019 to May 2022 were consecutively enrolled. Clinical, demographic, and concomitant medication data were collected. The endpoint was defined as OAC discontinuation for more than 30 days or a switch to an alternative therapy. Predictors of OAC non-persistence were investigated using a multivariable Cox regression model.Results: This study included 560 participants (51.1% men, mean age 80.9± 0.2 years). During a median follow-up of 20 months, medication persistence was observed in 322 patients (57.5%). Non-persistence was found to be significantly higher with warfarin than with NOAC (48.8% vs 33.6%, p = 0.006). In the multivariate analysis, OAC non-persistence was independently predicted by a history of permanent pacemaker implantation, the use of antiplatelet drugs, employee Medicare, living with children, college degree or above, and persistent AF (HR = 1.580, 1.586, 0.604, 0.668, 0.028, 0.769, p < 0.05, respectively). Treatment discontinuation within 3 months of discharge was observed in a large number of patients (81.8%). Medication discontinuation due to bleeding was more frequently observed in patients who continued for longer than 3 months (p < 0.001), while discontinuation due to patient preference was more frequent in those with shorter durations (≤ 3 months) (p = 0.049). Patient preference was the second leading cause of non-persistence in patients, regardless of whether they were taking warfarin or NOAC.Conclusion: OAC non-persistence remains high among elderly AF patients during long-term follow-up, with a significant proportion discontinuing shortly after discharge. This pattern of non-persistence is heavily influenced by demographic factors and patient preference. Further interventions should be developed based on the reasons and risk factors to improve persistence and initiated early in the treatment process.Keywords: atrial fibrillation, the elderly, oral anticoagulants, non-persistence

Guo-Yi Shen,1,&ast; Peng-Jie Yang,2,&ast; Wen-Shan Zhang,1 Jun-Biao Chen,1 Qin-Yong Tian,1 Yi Zhang,1 Bater Han2 1Department of Cardiothoracic Surgery, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou, 363000, People’s Republic of China; 2Department of Thoracic Surgery, Inner Mongolia Cancer Hospital & Affiliated People’s Hospital of Inner Mongolia Medical University, Huhhot, Inner Mongolia Autonomous Region, 010020, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yi Zhang, Department of Cardiothoracic Surgery, Zhangzhou Affiliated Hospital of Fujian Medical University, No. 59, Shengli West Road, Zhangzhou Xiangcheng District, Zhangzhou, 363000, People’s Republic of China, Tel +86-13906969033, Email zhangyi625@126.com Bater Han, Department of Thoracic Surgery, Inner Mongolia Cancer Hospital & Affiliated People’s Hospital of Inner Mongolia Medical University, No. 42, Zhaowuda Road, Saihan District, Huhhot, Inner Mongolia Autonomous Region, 010020, People’s Republic of China, Tel +86-13948120033, Email baterhanhbb@126.comBackground: Dysregulation of lipid metabolism is common in cancer. However, the molecular mechanism underlying lipid metabolism in esophageal squamous cell carcinoma (ESCC) and its effect on patient prognosis are not well understood. The objective of our study was to construct a lipid metabolism-related prognostic model to improve prognosis prediction in ESCC.Methods: We downloaded the mRNA expression profiles and corresponding survival data of patients with ESCC from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. We performed differential expression analysis to identify differentially expressed lipid metabolism-related genes (DELMGs). We used Univariate Cox regression and least absolute shrinkage and selection operator (LASSO) analyses to establish a risk model in the GEO cohort and used data of patients with ESCC from the TCGA cohort for validation. We also explored the relationship between the risk model and the immune microenvironment via infiltrated immune cells and immune checkpoints.Results: The result showed that 132 unique DELMGs distinguished patients with ESCC from the controls. We identified four genes (ACAA1, ACOT11, B4GALNT1, and DDHD1) as prognostic gene expression signatures to construct a risk model. Patients were classified into high- and low-risk groups as per the signature-based risk score. We used the receiver operating characteristic (ROC) curve and the Kaplan-Meier (KM) survival analysis to validate the predictive performance of the 4-gene signature in both the training and validation sets. Infiltrated immune cells and immune checkpoints indicated a difference in the immune status between the two risk groups.Conclusion: The results of our study indicated that a prognostic model based on the 4-gene signature related to lipid metabolism was useful for the prediction of prognosis in patients with ESCC.Keywords: biomarkers, esophageal squamous cell carcinoma, immune microenvironment, lipid metabolism, prognosis

Xiao Han,1 Qianjuan Zhang,2 Chengcheng Wang,3 Bingjuan Han4 1Department of Pediatrics, Jining First People’s Hospital, Jining, Shandong, 272011, People’s Republic of China; 2Department of Children’s Medical Rehabilitation Center, Jinan Maternity and Child Care Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, 250001, People’s Republic of China; 3Department of Pediatric Surgery, Jining First People’s Hospital, Jining, Shandong, 272011, People’s Republic of China; 4Department of Children’s Health Prevention, The Second Children & Women’s Healthcare of Jinan City, Jinan, Shandong, 271100, People’s Republic of ChinaCorrespondence: Bingjuan Han, Email hbj208@163.comPurpose: Mowat-Wilson syndrome (MWS) is an autosomal dominant disease caused by a pathogenic variant of the ZEB2 gene. The main clinical manifestations include special facial features, Hirschsprung disease (HSCR), global developmental delay and other congenital malformations. Here, we summarize the clinical characteristics and genetic mutation analysis of three Chinese patients with MWS.Patients and Methods: The clinical characteristics of the patients were monitored and the treatment effect was followed up. DNA was extracted from peripheral blood and analyzed by sequencing. Whole exome sequencing was then performed.Results: Three novel ZEB2 gene mutations were identified in 3 patients (c.1147_1150dupGAAC, p.Q384Rfs&ast;7, c.1137_1146del TAGTATGTCT, p.S380Nfs &ast;13 and c.2718delT, p.A907Lfs&ast;23). They all had special facial features, intellectual disability, developmental delay, microcephaly, structural brain abnormalities and other symptoms. After long-term regular rehabilitation treatment, the development quotient of each functional area of the patient was slightly improved.Conclusion: Our study expanded the mutation spectrum of ZEB2 and enriched our understanding of the clinical features of MWS. It also shows that long-term standardized treatment is of great significance for the prognosis of patients.Keywords: Mowat-Wilson syndrome, ZEB2, gene mutation, rehabilitation treatment, long-term follow-up

Zheyu Zhou,1,&ast; Shuya Cao,2,&ast; Chaobo Chen,3,4,&ast; Jun Chen,5 Xiaoliang Xu,6 Yang Liu,4 Qiaoyu Liu,6 Ke Wang,2 Bing Han,4 Yin Yin6 1Department of General Surgery, Nanjing Drum Tower Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Graduate School of Peking Union Medical College, Nanjing, 210008, People’s Republic of China; 2Hepatobiliary Center, The First Affiliated Hospital of Nanjing Medical University; Key Laboratory of Liver Transplantation, Chinese Academy of Medical Science; NHC Key Laboratory of Living Donor Liver Transplantation (Nanjing Medical University), Nanjing, 210029, People’s Republic of China; 3Department of General Surgery, Xishan People’s Hospital of Wuxi City, Wuxi, 214105, People’s Republic of China; 4Department of Hepatobiliary and Transplantation Surgery, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, 210008, People’s Republic of China; 5Department of Pathology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, 210008, People’s Republic of China; 6Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yin Yin, Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, No. 218, Jixi Road, Hefei, 230022, People’s Republic of China, Email jyinjyin@sina.com Bing Han, Department of Hepatobiliary and Transplantation Surgery, The Affiliated Drum Tower Hospital of Nanjing University Medical School, No. 321, Zhongshan Road, Nanjing, 210008, People’s Republic of China, Email hanbing_nju@163.comBackground: Edmondson-Steiner (E-S) grade is a pathological indicator of the degree of hepatocellular carcinoma (HCC) differentiation, and E-S grade III–IV is a poor prognostic factor for HCC patients. Predicting poorly differentiated HCC has essential significance for clinical decision-making. Although some studies have developed predictive models based on magnetic resonance imaging (MRI) and radiomics, radiomic features that require specific software for analysis are impractical for clinical work. This study aims to develop a novel and user-friendly nomogram model to predict E-S grade III–IV.Patients and Methods: Medical data on patients meeting the inclusion criteria were obtained from the Nanjing Drum Tower Hospital HCC database (January 2020 to December 2022). Univariate analysis was used to screen for risk factors associated with E-S grade III–IV. A novel nomogram was established based on the subsequent multivariate logistic regression analysis. The performance of the established model was evaluated through diagnostic ability, calibration, and clinical benefits.Results: Overall, 240 HCC patients were included in this study. Among them, 103 were highly differentiated (E-S grade I–II) HCC and 137 were poorly differentiated (E-S grade III–IV) HCC. A nomogram model that integrated alpha-fetoprotein (AFP), des-γ-carboxy prothrombin (DCP), hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibodies (HCVAb), aspartate aminotransferase to lymphocyte ratio index (ALRI), and macrovascular invasion was established. The novel model had a good diagnostic performance with an area under the curve (AUC) value of 0.763. Meanwhile, the model had a diagnostic accuracy of 72.5%, a sensitivity of 78.1%, and a specificity of 65.1%. The calibration curve showed good calibration of the nomogram model (mean absolute error = 0.043), and the decision curve analysis (DCA) demonstrated that the clinical benefit was provided.Conclusion: Our developed nomogram model could successfully predict E-S grade III–IV in HCC patients, which may be helpful in clinical decision-making.Keywords: hepatocellular carcinoma, Edmondson-Steiner grade, nomogram, prediction

Wenjuan Yang,1,2 Bing Han,1 Bei Zheng,1 Ying Li,1 Tiefei Yao,3 Mei Han,3 Gonghua Li,1 Meiling Zhang1 1Department of Pharmacy, Tongde Hospital of ZheJiang Province, Hangzhou, Zhejiang, People’s Republic of China; 2Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, Zhejiang, People’s Republic of China; 3Department of Geriatric Medicine, Tongde Hospital of ZheJiang Province, Hangzhou, Zhejiang, People’s Republic of ChinaCorrespondence: Gonghua Li; Meiling Zhang, Department of Pharmacy, Tongde Hospital of ZheJiang Province, Hangzhou, Zhejiang, People’s Republic of China, Tel +86-0571-89972239, Email ligonghua88@163.com; zml9998@sina.comAbstract: Coronavirus disease 2019 (COVID-19) emergence in late 2019, and wide spread quickly in the world. In China, the COVID-19 epidemic situation entered a low level now. With the arrival of flu season, the number of patients with respiratory symptoms is increasing. We reported three cases of patients who co-infected with SARS-CoV-2 and influenza A virus (IAV), and they were all treated with nirmatrelvir-ritonavir (NMV/r) and baloxavir marboxil. Due to the overlapping clinical features between the two diseases, it is important to identified them and gave the antiviral therapy timely.Keywords: COVID-19, influenza, co-infection, baloxavir marboxil, nirmatrelvir-ritonavir

Xiaoyu Pan,1,&ast; Huazheng Zhu,2,&ast; Jinglong Du,1 Guangtao Hu,1 Baoru Han,1 Yuanyuan Jia1 1College of Medical Informatics, Chongqing Medical University, Chongqing, People’s Republic of China; 2College of Intelligent Technology and Engineering, Chongqing University of Science and Technology, Chongqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuanyuan Jia, Email yyjia@cqu.edu.cnAim: The Coronavirus Disease 2019 (COVID-19) pandemic has increased the public health burden and brought profound disaster to humans. For the particularity of the COVID-19 medical images with blurred boundaries, low contrast and different infection sites, some researchers have improved the accuracy by adding more complexity. Also, they overlook the complexity of lesions, which hinder their ability to capture the relationship between segmentation sites and the background, as well as the edge contours and global context. However, increasing the computational complexity, parameters and inference speed is unfavorable for model transfer from laboratory to clinic. A perfect segmentation network needs to balance the above three factors completely. To solve the above issues, this paper propose a symmetric automatic segmentation framework named MS-DCANet. We introduce Tokenized MLP block, a novel attention scheme that uses a shift-window mechanism to conditionally fuse local and global features to get more continuous boundaries and spatial positioning capabilities. It has greater understanding of irregular lesion contours. MS-DCANet also uses several Dual Channel blocks and a Res-ASPP block to improve the ability to recognize small targets. On multi-modality COVID-19 tasks, MS-DCANet achieved state-of-the-art performance compared with other baselines. It can well trade off the accuracy and complexity. To prove the strong generalization ability of our proposed model, we apply it to other tasks (ISIC 2018 and BAA) and achieve satisfactory results.Patients: The X-ray dataset from Qatar University which contains 3379 cases for light, normal and heavy COVID-19 lung infection. The CT dataset contains the scans of 10 patient cases with COVID-19, a total of 1562 CT axial slices. The BAA dataset is obtained from the hospital and includes 387 original images. The ISIC 2018 dataset is from the International Skin Imaging Collaborative (ISIC) containing 2594 original images.Results: The proposed MS-DCANet achieved evaluation metrics (MIOU) of 73.86, 97.26, 89.54, and 79.54 on the four datasets, respectively, far exceeding other current state-of-the art baselines.Conclusion: The proposed MS-DCANet can help clinicians to automate the diagnosis of COVID-19 patients with different symptoms.Keywords: multi-modality COVID-19 lesion segmentations, laboratory to clinic, multi-scale feature learning, depthwise separable convolution

Ling Sun,1,&ast; Bing Han,2,&ast; Yu Wang,1,&ast; Wenwu Zhu,2 Jianguang Jiang,1 Ailin Zou,1 Boyu Chi,1,3 Lipeng Mao,1,3 Yuan Ji,1 Qingjie Wang,1 Liming Tang4 1Department of Cardiology, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, 213003, People’s Republic of China; 2Department of Cardiology, Xuzhou Central Hospital, Xuzhou Clinical School of Nanjing Medical University, Xuzhou, Jiangsu, 221009, People’s Republic of China; 3Dalian Medical University, Dalian, Liaoning, 116000, People’s Republic of China; 4Department of Gastrointestinal Disease, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, 213003, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Liming Tang, Center of Gastrointestinal Disease, Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, 29 Xinglong Alley, Changzhou, Jiangsu, 213003, People’s Republic of China, Email drtangliming@163.com Yuan Ji, Department of Cardiology, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, 213003, People’s Republic of China, Email jiyuan1213@aliyun.comObjective: In this study, a risk score for ventricular arrhythmias (VA) were evaluated for predicting the risk of ventricular arrhythmia (VA) of acute myocardial infarction (AMI) patients.Methods: Patients with AMI were divided into two sets according to whether VA occurred during hospitalization. Another cohort was enrolled for external validation. The area under the curve (AUC) of receiver operating characteristic (ROC) was calculated to evaluate the accuracy of the model.Results: A total of 1493 eligible patients with AMI were enrolled as the training set, of whom 70 (4.7%) developed VA during hospitalization. In-hospital mortality was significantly higher in the VA set than in the non-VA set (31.4% vs 2.7%, P=0.001). The independent predictors of VA in patients with AMI including Killip grade ≥ 3, STEMI patients, LVEF < 50%, frequent premature ventricular beats, serum potassium < 3.5 mmol/L, type 2 diabetes, and creatinine level. The AUC of the model for predicting VT/VF in the training set was 0.815 (95% CI: 0.763– 0.866). A total of 1149 cases were enrolled from Xuzhou Center Hospital as the external validation set. The AUC of the model in the external validation set for predicting VT/VF was 0.755 (95% CI: 0.687– 0.823). Calibration curves indicated a good consistency between the predicted and the observed probabilities of VA in both sets.Conclusion: We have established a clinical prediction risk score for predicting the occurrence of VA in AMI patients. The prediction score is easy to use, performs well and can be used to guide clinical practice.Keywords: ventricular tachycardia, ventricular flutter and fibrillation, risk stratification, scoring system, acute myocardial infarction

Jun Lu,1– 4,&ast; Yuqing Lou,1,&ast; Yanwei Zhang,1,&ast; Runbo Zhong,1 Wei Zhang,1 Xueyan Zhang,1 Huimin Wang,1 Tianqing Chu,1 Baohui Han,1– 3 Hua Zhong1,3 1Department of Pulmonary Medicine, Shanghai Chest Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 2Shanghai Institute of Thoracic Oncology, Shanghai Chest Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 3Translational Medical Research Platform for Thoracic Oncology, Shanghai Chest Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 4Department of Bio-Bank, Shanghai Chest Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Baohui Han; Hua Zhong, Department of Pulmonary Medicine, Shanghai Chest Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200030, People’s Republic of China, Email xkyyhan@gmail.com; 18930858216@163.com; eddiedong8@hotmail.comBackground: Nanocarrier platforms have been indicated to have great potential in clinical practice to treat non-small cell lung cancer (NSCLC). Our previous Phase III clinical study revealed that polymeric micellar paclitaxel (Pm-Pac) is safe and efficacious in advanced NSCLC patients. However, the histopathological-toxicological profile of Pm-Pac in mammals remains unclear.Methods: We examined the Pm-Pac-induced antitumour effect in both A549/H226 cells and A549/H226-derived xenograft tumour models.. And then, we evaluated the short-term and long-term toxicity induced by Pm-Pac in healthy Sprague‒Dawley (SD) rats. The changes in body weight, survival, peripheral neuropathy, haematology, and histopathology were studied in SD rats administered Pm-Pac at different dosages.Results: In the A549-derived xenograft tumour model, better therapeutic efficacy was observed in the Pm-Pac group than in the solvent-based paclitaxel (Sb-Pac) group when an equal dosage of paclitaxel was administered. Toxicity assessments in healthy SD rats indicated that Pm-Pac caused toxicity at an approximately 2- to 3-fold greater dose than Sb-Pac when examining animal body weight, survival, peripheral neuropathy, haematology, and histopathology. Interestingly, based on histopathological examinations, we found that Pm-Pac could significantly decrease the incidences of paclitaxel-induced brain and liver injury but could potentially increase the prevalence of paclitaxel-induced male genital system toxicity.Conclusion: This study introduces the toxicological profile of the engineered nanoparticle Pm-Pac and provides a novel perspective on the Pm-Pac-induced histopathological-toxicological profile in a rat model.Graphical Abstract: Keywords: nanomedicine, polymeric micellar paclitaxel, NSCLC, toxicological profile

Hao Wang, Xin Yin, Tianyi Fang, Shenghan Lou, Bangling Han, Jialiang Gao, Yufei Wang, Daoxu Zhang, Xibo Wang, Zhanfei Lu, Junpeng Wu, Jiaqi Zhang, Yimin Wang, Yao Zhang, Yingwei Xue Department of Gastroenterological Surgery, Harbin Medical University Cancer Hospital, Harbin, 150081, People’s Republic of ChinaCorrespondence: Yingwei Xue, Department of Gastroenterological Surgery, Harbin Medical University Cancer Hospital, Harbin, 150081, People’s Republic of China, Email xueyingwei@hrbmu.edu.cnBackground: Aging has a negative impact on the immune function of patients. The purpose of this study was to construct an age-related specific immune index according to the immune aging phenomenon of gastric cancer (GC) and explore its prognostic value.Methods: This study retrospectively analyzed patients who underwent radical GC surgery in the Department of Gastrointestinal Surgery, Affiliated Cancer Hospital of Harbin Medical University, from August 2014 to December 2016 and divided them into a training cohort and a validation cohort. A new immune score, the GC-specific immune index (GSII), was developed as a series of lymphocyte subsets associated with the prognosis of patients with GC. Then, the receiver operating characteristic (ROC) curve was used to compare the prediction performance. The Kaplan‒Meier method and Log rank test were used to analyze the overall survival of patients. Cox hazard regression models were used to identify independent risk factors associated with prognosis. Finally, a nomogram model was constructed by combining the GSII and clinicopathological characteristics, and the calibration chart, consistency index, and decision curve were used to test the performance of the model.Results: Aging did not significantly affect CD8 cell counts but decreased CD4 and CD19 cell counts. Based on the Cox analysis, the GSII of patients ≤ 60 years old was 0.079×lg CD4+0.348×lg CD19, and the GSII of patients > 60 years old was 0.058×lg CD4. A decreased GSII was indicative of a poor prognosis and was an independent risk factor associated with patient outcomes. The nomogram constructed based on the GSII and clinicopathological features accurately predicted patient prognosis. Furthermore, the GSII was well validated in the validation cohort.Conclusion: The GSII constructed for the special immune aging phenomenon of GC can accurately predict patient prognosis.Keywords: gastric cancer, lymphocyte subsets, age, prognosis

Yuan Zhang,1,&ast; Hongli Yue,1,&ast; Yirui Qin,1 Jiajing Wang,1 Chenyang Zhao,1 Miao Cheng,1 Bo Han,2 Ruquan Han,1 Weihua Cui1,&ast; 1Department of Anesthesiology, Beijing Tian Tan Hospital, Capital Medical University, Beijing, People’s Republic of China; 2Department of Neurosurgery, Beijing Tian Tan Hospital, Capital Medical University, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ruquan Han; Weihua Cui, Department of Anesthesiology, Beijing Tian Tan Hospital, Capital Medical University, No. 119, Nan Si Huan Xi Lu, Fengtai District, Beijing, 100070, People’s Republic of China, Tel +8613701285393 ; Tel +8613701285393, Fax +861059976658, Email ruquan.han@ccmu.edu.cn; weihuacui@ccmu.edu.cnPurpose: Patients undergoing intraspinal tumor resection usually experience severe postoperative pain. Inadequate postoperative analgesia usually leads to severe postsurgical pain, which could cause patients to suffer from many other related complications. Recently, an increasing number of studies have found that gabapentin can relieve hyperalgesia, postoperative pain, and postoperative inflammation. However, there have been no reports on the use of gabapentin combined with sufentanil preoperatively for acute pain following intraspinal tumor resection.Study Design and Methods: This is a protocol for a randomized, placebo-controlled, and double-blinded trial. One-hundred and sixty-eight participants with chronic pain related to the intraspinal tumor will be randomized into the gabapentin and placebo groups in a 1:1 ratio. In the gabapentin group, patients will be given 300 mg gabapentin orally 36 h, 24 h, and 12 h before surgery; the placebo group will receive a placebo orally at the same time points preoperatively. To estimate the efficacy and safety endpoints, all the researchers and patients will be blinded until the completion of this study. The primary outcome will be the consumption of sufentanil within 48 h postoperatively. The secondary outcomes include the visual analog scale pain score and Von Frey mechanical pain threshold 36 h and 24 h before and 24 h and 48 h after surgery, the incidence of postoperative nausea, vomiting, and drowsiness, the length of hospital stay and medical expenses.Discussion: This trial is to evaluate the efficacy and safety of gabapentin combined with sufentanil for postoperative analgesia in patients who complain of pain before intraspinal tumor resection. The findings will provide a new strategy for multimode perioperative analgesia management in these patients.Keywords: gabapentin, intraspinal tumor resection, multimode perioperative analgesia, opiates

Chunxue He,1,2,&ast; Mingqiang Gong,3,4,&ast; Gengxiao Li,1,2 Yunxia Shen,2 Longyin Han,5 Bin Han,6 Mingwu Lou2 1Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, People’s Republic of China; 2Department of Radiology, Longgang District Central Hospital of Shenzhen, Shenzhen, People’s Republic of China; 3The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, People’s Republic of China; 4Department of Acupuncture, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, People’s Republic of China; 5Department of Neurology, Beijing Longfu Hospital, Beijing, People’s Republic of China; 6Department of Rehabilitation Medicine, Longgang District Central Hospital of Shenzhen, Guangdong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bin Han; Mingwu Lou, Tel +86 13928488998 ; +86 13808854650, Email hhbb08@163.com; mingwulou@sina.comPurpose: To investigate white matter alterations in post-stroke cognitive impairment (PSCI) patients at the subacute stage employing diffusion kurtosis and tensor imaging.Methods: Thirty PSCI patients at the subacute phase and 30 healthy controls (HC) underwent diffusion kurtosis imaging (DKI) scans and neuropsychological assessments. Based on the tract-based spatial statistics and atlas-based ROI analysis, fractional anisotropy (FA), mean diffusivity (MD), mean kurtosis (MK), kurtosis fractional anisotropy (KFA), axial kurtosis (AK), and radial kurtosis (RK) were compared in specific white matter fiber bundles between the groups (with family-wise error correction). Adjusting for age and gender, a partial correlation was conducted between neurocognitive assessments and DKI metrics in the PSCI group.Results: In comparison with the HC, PSCI patients significantly showed decreased MK, RK, and FA and increased MD values in the genu of corpus callosum, anterior limb internal capsule, and left superior corona radiata. In addition, DKI detected more white matter region changes in MK (31/48), KFA (40/48), and RK (25/48) than DTI with FA (28/48) and MD (21/48), which primarily consisted of the right cingulum, right superior longitudinal fasciculus, and left posterior limb of internal capsule. In the left anterior limb of internal capsule, MK and RK values were significantly negatively correlated with TMT-B (r = − 0.435 and − 0.414, P < 0.05), and KFA values (r = − 0.385, P < 0.05) of corpus callosum negatively associated with TMT-B.Conclusion: Combing DTI, DKI, and neuropsychological tests, we found extensive damaged white matter microstructure and poor execution performance in subacute PSCI patients. DKI could detect more subtle white matter changes than DTI metrics. Our findings provide added information for exploring the mechanisms of PSCI and conducting cognitive rehabilitation in the subacute stage.Keywords: diffusion kurtosis imaging, diffusion tensor imaging, subacute ischemic stroke, cognitive impairment

Bin Han,1,&ast; Feng Zang,2,&ast; Juan Liu,2 Songqin Li,2 Weihong Zhang,2 Yongxiang Zhang,2 Zhanjie Li2 1Department of Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, Jiangsu, People’s Republic of China; 2Department of Infection Control, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, Jiangsu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yongxiang Zhang; Zhanjie Li, Tel +86-13951951616 ; +86-18052106999, Email zyx3019@sina.cn; LZJ070591@163.comPurpose: To improve the ability of infection prevention and control (IPC) of medical staff during the COVID-19 epidemic period, the “four-step” mode of whole staff training and assessment was used.Methods: During the period from March 9 to March 18, 2020, 5425 medical staff from The First Affiliated Hospital of Nanjing Medical University were selected as the objects of this study. There are four stages in the training assessment mode. The first stage is the basic assessment stage; the second stage releases the electronic version of the knowledge point manual; the third stage conducts online exercises; the fourth stage conducts the final assessment.Results: In the first stage, the participation rate of medical staff was 95.04%. In the fourth stage, the participation rate of medical staff was 98.01%. The average score of female medical staff in the first stage and the fourth stage was higher than that of males (P< 0.001). The average score of medical staff under 30 years old in the first stage and the fourth stage was higher than that of other age groups (P< 0.05). In the fourth stage, the correct rate of each part of exercises in the knowledge points of IPC was higher than that in the first stage (P< 0.001). In the two stages, the two parts of “COVID-19 prevention and control” and “multi-drug resistant bacteria prevention and control” had the highest accuracy, while the “disinfection and sterilization” and “infectious disease management” had the lowest accuracy (P< 0.001).Conclusion: The “four-step” infection control training assessment mode has realized “full participation” and “effective training”, and the level of medical staff’s IPC has been significantly improved.Keywords: training and assessment tool, infection prevention and control, COVID-19, four-step, questionnaire

Hai-Juan Zhu,1,2,&ast; Yan He,3,&ast; Sheng-You Wang,2 Bo Han,2 Ye Zhang1 1Department of Anesthesiology and Perioperative Medicine, The Second Hospital of Anhui Medical University, Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education Institutes, Anhui Medical University, Hefei, 230601, People’s Republic of China; 2Department of Anesthesiology, Anhui Maternal and Child Health Care Hospital, Maternal and Child Health Care Hospital of Anhui Medical University, Hefei, 230001, People’s Republic of China; 3Department of Anesthesiology, Wannan Medical College, Wuhu, Anhui Province, 241002, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ye ZhangDepartment of Anesthesiology and Perioperative Medicine, The Second Hospital of Anhui Medical University, Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education Institutes, Anhui Medical University, Hefei, 230601, People’s Republic of China, Tel +86-551-63869485; +86-13966768081, Fax +86 551-63869400, Email zhangy@ahmu.edu.cnPurpose: This study aimed to explore the efficacy and safety of chloroprocaine for activating labor analgesia and the optimal concentration compared to lidocaine.Patients and Methods: Ninety-six nulliparous parturients were randomly assigned to three groups: LD group, patients received the conventional initial dose of 6 mL of 1% lidocaine; CP1.5 group, patients received 6 mL of 1.5% chloroprocaine as the initial dose; and CP1.2 group, patients received 7.5 mL of 1.2% chloroprocaine as initial dose. Labor analgesia was maintained in all patients via a programmed intermittent epidural bolus (PIEB). The primary outcome was the analgesia onset time. Secondary outcomes included the visual analog scale (VAS) scores, the interval and duration of uterine contractions during the first 12 contractions, failure to reach adequate analgesia, labor and neonatal outcomes, maternal satisfaction and adverse effects.Results: Parturients in the CP1.5 and CP1.2 groups achieved a shorter onset time than those in the LD group (hazard ratio (HR) = 6.540; 95% confidence interval (CI), 3.503– 12.210; P < 0.001 and HR = 3.460; 95% CI, 1.905– 6.282; P < 0.001, respectively). The median time (95% CIs) to adequate analgesia was 12.0 (10.9– 13.1), 7.0 (6.2– 7.8) and 8.0 (7.5– 8.5) minutes in the LD, CP1.5 and CP1.2 groups, respectively. PIEB in the CP1.5 group was associated with lower VAS scores, patient-controlled epidural analgesia (PCEA) boluses, and analgesic consumption; a shorter time from epidural initiation to the first PCEA demand; and higher maternal satisfaction scores than the other two groups (P < 0.01). The interval and duration of uterine contractions, labor and newborn outcomes and adverse effects were comparable among the three groups.Conclusion: Chloroprocaine provided a faster onset of labor analgesia than lidocaine. Thus, 6 mL of 1.5% chloroprocaine might be a superior volume and concentration regimen to 7.5 mL of 1.2% chloroprocaine for activating labor analgesia.Clinical Trial Registration Statement: The study was registered prior to subject enrollment at www.chictr.org.cn (ChiCTR2100049113).Keywords: epidural, labor analgesia, onset time, activation, chloroprocaine, lidocaine

Boyue Han,1,2,&ast; Zhangyuan Gu,3,&ast; Zhebin Liu,1,2 Hong Ling1,2 1Department of Breast Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, 200032, People’s Republic of China; 2Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, People’s Republic of China; 3Department of Breast Surgery, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, 200040, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhebin Liu; Hong Ling, Email zhebinliu@shca.org.cn; linghong98@aliyun.comPurpose: The objective of this study was to compare the demographic characteristics, clinicopathological factors and survival outcomes between infiltrating lobular carcinoma (ILC) and infiltrating ductal carcinoma (IDC) using our single-center database.Methods: Seventeen thousand two hundred and three breast cancer patients were treated at Fudan University Shanghai Cancer Center (FUSCC) from January 2000 to December 2017. We identified 365 cases with ILC and 16,838 cases with IDC. The Pearson chi-square test was used to compare tumor characteristics, and the Kaplan–Meier methods were used to perform the survival analysis.Results: ILC had some distinctive characteristics from IDC such as older age (ranged from 61 to 80: ILC 26.8% vs IDC 19.9%, P < 0.001; over 80: ILC 1.6% vs IDC 0.8%, P < 0.001), larger tumor size (ranged from 2 to 5: ILC 45.2% vs IDC 37.1%, P = 0.011), much more hormone receptor expression (ILC 92.9% vs IDC 73.0%, P < 0.001), extremely less HER-2 expression (ILC 7.1% vs IDC 25.9%, P < 0.001). The overall survival and disease-free survival of ILC were worse than IDC (5-year OS, ILC 93.6% vs IDC 94.5%, P < 0.001; 5-year DFS, ILC 88.5% vs IDC 91.6%, P = 0.008). It was worth noting that the ILC patients had a worse overall survival than IDC patients after our propensity score matching study (P = 0.037). The univariate analysis concluded that positive HR (hormone receptor), high expression of Ki-67 and higher pathologic tumor stage were poor prognostic markers of ILC. Multivariate analysis demonstrated that tumor stage was a poor prognostic marker after adjustment for the effects of the above three factors. The most common primary site of metastasis was bone, but the proportion in the ILC group was much higher than that in the IDC group (56.25% vs 36.40%, P = 0.003).Conclusion: Compared with IDC, ILC survived worse and was more prone to bone metastasis. Therefore, a comprehensive understanding of ILC and specific treatments are needed for further research.Keywords: infiltrating lobular carcinoma, infiltrating ductal carcinoma, survival, prognosis

Hongtao Ding, Yong Hai, Yuzeng Liu, Li Guan, Aixing Pan, Xinuo Zhang, Bo Han, Yue Li, Peng Yin Department of Spine Surgery, Beijing Chao-Yang Hospital, Capital Medical University of China, Beijing, People’s Republic of ChinaCorrespondence: Yong Hai; Yuzeng Liu, Department of Spine Surgery, Beijing Chao-Yang Hospital, Capital Medical University of China, Beijing, People’s Republic of China, Tel +86 10-85231229 ; +86 13801221889 ; +86 13811552832, Email prof.haiyong@yahoo.com; yong.hai@ccmu.edu.cn; beijingspine2010@163.comStudy Design: This was a prospective randomized controlled trial study.Objective: To elucidate clinical and radiographic outcomes and complications of cortical bone trajectory (CBT)-screw fixation in patients with osteoporosis at 24-month follow-up and to compare the results with those after transforaminal lumbar interbody fusion (TLIF) using traditional pedicle screw (PS) fixation.Methods: We enrolled 124 patients and randomly assigned them to two groups (each group had 62 participants). The primary outcome was fusion rate. Secondary outcomes were VAS, Oswestry Disability Index (ODI), and Japanese Orthopaedic Association (JOA) scores, operation duration, incision length, estimated blood loss, drainage volume, radiological outcomes, and complications.Results: At the 6- and 12-month follow-up points, similar fusion rates were observed based on CT scans in both groups (P=0.583 and 0.583). CBT provided significantly better short-term functional status at 3 months postoperation on ODI and JOA scores (P=0.012 and 0) and similar improvements in pain intensity and functional status at other follow-up points. In addition, CBT resulted in significantly better surgical characteristics. Notably, CBT fixation led to lower incidence of screw loosening (P=0.006).Conclusion: CBT-screw fixation for single-level lumbar fusion in patients with osteoporosis provided improvement in clinical symptoms comparable to that of TLIF using PS fixation. Significantly better lumbar stability was found in the CBT group. We suggest that CBT-screw fixation is a reasonable and superior alternative to PS in TLIF in osteoporosis.Trial Registration Number: ChiCTR1900022658.Date of Registration: April 20, 2019.Keywords: cortical bone-trajectory screw, pedicle screw, lumbar fusion, osteoporosis, randomized controlled trial

Abstract Our goal is to explore quantitative motor features in critically ill patients with severe brain injury (SBI). We hypothesized that computational decoding of these features would yield information on underlying neurological states and outcomes. Using wearable microsensors placed on all extremities, we recorded a median 24.1 (IQR: 22.8–25.1) hours of high-frequency accelerometry data per patient from a prospective cohort (n = 69) admitted to the ICU with SBI. Models were trained using time-, frequency-, and wavelet-domain features and levels of responsiveness and outcome as labels. The two primary tasks were detection of levels of responsiveness, assessed by motor sub-score of the Glasgow Coma Scale (GCSm), and prediction of functional outcome at discharge, measured with the Glasgow Outcome Scale–Extended (GOSE). Detection models achieved significant (AUC: 0.70 [95% CI: 0.53–0.85]) and consistent (observation windows: 12 min–9 h) discrimination of SBI patients capable of purposeful movement (GCSm > 4). Prediction models accurately discriminated patients of upper moderate disability or better (GOSE > 5) with 2–6 h of observation (AUC: 0.82 [95% CI: 0.75–0.90]). Results suggest that time series analysis of motor activity yields clinically relevant insights on underlying functional states and short-term outcomes in patients with SBI.

Hui-Ling Chen,1,2,&ast; Yi-Bin Zeng,3,&ast; Zheng-Yan Zhang,1,2,&ast; Chao-Yue Kong,1,2 Shi-Long Zhang,1,2 Zhan-Ming Li,1,2 Jia-Ting Huang,1,2 Ya-Yun Xu,1,2 Yu-Qin Mao,1,2 Pei-Ran Cai,1,2 Bing Han,1,2 Wu-Qing Wang,3 Li-Shun Wang1,2 1Center for Traditional Chinese Medicine and Gut Microbiota, Minhang Hospital, Fudan University, Shanghai, 201100, People’s Republic of China; 2Institute of Fudan-Minhang Academic Health System, Minhang Hospital, Fudan University, Shanghai, 201100, People’s Republic of China; 3Dermatological Department, Minhang Hospital, Fudan University, Shanghai, 201100, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Li-Shun Wang; Wu-Qing WangCenter for Traditional Chinese Medicine and Gut Microbiota, Minhang Hospital, Fudan University, Shanghai, 201100, People’s Republic of ChinaTel +86 21-64923400-6244Email lishunwang@fudan.edu.cn; wuqingw2006@sina.comBackground: Psoriasis is a chronic autoinflammatory skin disease, and its aetiology remains incompletely understood. Recently, gut microbial dysbiosis is found to be tightly associated with psoriasis.Objective: We sought to reveal the causal role of gut microbiota dysbiosis in psoriasis pathogenesis and investigate the protective effect of healthy commensal bacteria against imiquimod -induced psoriasis-like skin response.Methods: By using fecal microbial transplantation (FMT), 16S rRNA gene-based taxonomic profiling and Lactobacillus supplement, we have assessed the effect of FMT from healthy individuals on psoriasis-like skin inflammation and associated immune disorders in imiquimod-induced psoriasis mice.Results: Here, by using psoriasis mice humanized with the stools from healthy donors and psoriasis patients, the imiquimod-induced psoriasis in mice with psoriasis patient stool was found to be significantly aggravated as compared to the mice with healthy donor stools. Further analysis showed fecal microbiota of healthy individuals protected against Treg/Th17 imbalance in psoriasis. Moreover, we found the gut and skin microbiome in mice receipted with gut microbiota of healthy individuals (HD) differed from those of mice receipted with gut microbiota of psoriasis patients (PSD). 16S rRNA sequencing revealed that Lactobacillus reuteri was greatly enriched in fecal and cutaneous microbiome of HD mice as compared to PSD mice. Intriguingly, supplement with Lactobacillus reuteri was sufficient to increase the expression of anti-inflammatory gene IL-10, reduce Th17 cells counts and confer resistance to imiquimod-induced inflammation on the mice with gut microbiota dysbiosis.Conclusion: Our results suggested that the gut microbiota dysbiosis is the potential causal factor for psoriasis and the gut microbiota may serve as promising therapy target for psoriasis patients.Keywords: gut microbiota, cutaneous microbiome, psoriasis, fecal microbiota transplantation, Lactobacillus reuteri, Th17

Rong Qiao,1,&ast; Mengxia Li,2,&ast; Runbo Zhong,1 Yujie Wei,3 Jun Wang,3 Zheng Zhang,3 Ling Wang,3 Tian Xu,4 Yue Wang,1 Liping Dai,5 Wanjian Gu,4 Baohui Han,1 Rongxi Yang2,3 1Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai, 200030, People’s Republic of China; 2Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing, 210000, People’s Republic of China; 3Nanjing TANTICA Biotechnology Co. Ltd, Nanjing, 210000, People’s Republic of China; 4Department of Clinical Laboratory, Jiangsu Province Hospital of Chinese Medicine, Nanjing, 210000, People’s Republic of China; 5Henan Institute of Medical and Pharmaceutical Sciences, Academy of Medical Science, Zhengzhou University, Zhengzhou, 450052, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Rongxi Yang; Baohui Han Fax +86 25 86527613Email rongxiyang@njmu.edu.cn; 18930858216@163.comPurpose: Lung cancer (LC) brings great burden to the society worldwide. Exploring novel biomarkers in vitro for the early detection of LC would be of great importance.Patients and Methods: We measured DNA methylation levels of 21 CpG sites within Patatin-like phospholipase domain containing 2 (PNPLA2) gene in the peripheral blood of 168 early-stage LC cases (94.0% LC at stage I) and 187 age- and gender-matched cancer-free controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression adjusted for covariates. Non-parametric tests were applied for the comparisons of stratified groups.Results: Hypomethylation of PNPLA2_CpG_8,10 and hypermethylation of PNPLA2_CpG_9 were correlated to the early-stage LC with the ORs of 1.44 (95% CI: 1.06– 1.96, P = 0.018) and 0.82 (95% CI: 0.69– 0.98, P = 0.029), respectively. The associations were still significant for the very early-stage LC patients (stage I). Further gender- and age-stratified analyses indicated that the association between hypomethylation of PNPLA2_CpG_8,10 and LC existed only in females and in subjects younger than 55 years. In addition, the association between LC and hypermethylation of PNPLA2_CpG_6 and PNPLA2_CpG_9 was also observed in the younger population.Conclusion: Taken together, our study has proved the hypothesis that the altered methylation in the peripheral blood may be correlated with the burden of cancer at an early stage. Here, we find a novel association between blood-based aberrant PNPLA2 methylation and LC at a very early stage and particularly for women at a younger age.Keywords: lung cancer, DNA methylation, peripheral blood, biomarker

Feng Pan,1,&ast; Ai-Ting Lu,1,2,&ast; Xiaowei Mao,1,&ast; Yefeng Chen,3 Yizhuo Zhao,1 Baohui Han1 1Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 2Department of Nursing, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 3Department of Respiratory Medicine, Shaoxing People’s Hospital (Shaoxing Hospital, Zhejiang University School of Medicine), Shaoxing, Zhejiang, 312000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Baohui Han; Yizhuo ZhaoDepartment of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of ChinaEmail 18930858216@163.com; zhyz920916@126.comBackground: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a valuable tool for diagnosing pulmonary disease due to its efficiency and safety. We retrospectively analyzed patients with mediastinal masses who underwent diagnostic EBUS-TBNA at Shanghai Chest Hospital, and evaluated the clinical accuracy of EBUS-TBNA in the diagnosis mediastinal masses.Method: From 2009 and 2014, patients who received EBUS-TBNA to diagnose a isolated mediastinal mass were enrolled. Clinical follow-up was performed to ascertain the patient’s final diagnosis.Results: Forty-six patients were enrolled in this study. Thirty-seven were diagnosed with an oncologic disease, 3 were diagnosed with a mediastinal infection, and 2 were found to have a mediastinal goiter. The overall sensitivity, specificity, positive predictive value, negative predictive value, diagnostic yield was 63.6%, 100%, 100%, 42.9%, and 71.4%, respectively.Conclusion: EBUS-TBNA is a safe and effective means of diagnosing mediastinal masses.Keywords: EBUS-TBNA, mediastinum, diagnosis

Lin Wei,1 Xiao Han,2 Xue Li,1 Bingjuan Han,1 Wenying Nie1 1Jinan Maternity and Child Care Hospital Affiliated to Shandong First Medical University, Jinan Maternal and Child Care Hospital, Jinan, 250001, Shandong Province, People’s Republic of China; 2Jinan Central Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250012, Shandong Province, People’s Republic of ChinaCorrespondence: Bingjuan Han; Wenying Nie No. 2, Jianguo Xiaojing 3rd Road, Shizhong District, Jinan, Shandong Province, People’s Republic of ChinaTel/Fax +86 531-82060983; +86 531-8902957Email hbj208@163.com; goodddd@163.comPurpose: Mowat–Wilson syndrome (MWS) is a rare complex malformation syndrome which is characterized by typical facial dysmorphism, moderate to severe intellectual disability, global developmental delay, and multiple congenital anomalies. Here, we summarize the clinical characteristics and gene mutation analysis of a Chinese boy with MWS.Patients and Methods: The clinical features of the patient were monitored. DNA extracted from peripheral blood was subjected to sequencing analysis. Then, the whole-exome sequencing was performed.Results: A novel deletion mutation (c.1137_1146del TAGTATGTCT) was identified in exon 8 of the ZEB2 gene. The deletion mutation was predicted to produce a truncated protein (p.S380Nfs*13), resulting in haploinsufficiency. The patient presented with short stature, microcephaly, congenital heart defects, cryptorchidism, corpus callosum agenesis, global developmental delay, and intellectual disability. Furthermore, he demonstrated bilateral sensorineural hearing loss. This manifestation is less common in MWS. It is first reported in Chinese patients with MWS. Clinical follow-up showed that the facial features of MWS developed with time. The facial features of the patient were not obvious except for the uplifted ear lobes at the age of 3 months. At the age of 22 months, the facial characteristics of the patient included ocular hypertelorism, frontal bossing, rounded nasal tip, sparse eyebrows, prominent chin, widely spaced teeth, and uplifted ear lobes with a central depression.Conclusion: A novel deletion mutation of the ZEB2 gene was identified. This work contributes to expanding the mutation spectra of MWS. Our results may reflect the variability of the phenotype in MWS.Keywords: Mowat–Wilson syndrome, ZEB2, gene mutation, phenotype

Bo Han,1,&ast; Tao Wang,1,&ast; Zhigang Xue,2 Tao Wen,1 Ling Lu,3 Jie Meng,1 Jian Liu,1 Sizhu Wu,3 Jianchun Yu,2 Haiyan Xu1 1Department of Biomedical Engineering, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Beijing, People’s Republic of China; 2Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 3State Key Laboratory of Organic-Inorganic Composites, Beijing University of Chemical Technology, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Haiyan Xu; Jianchun Yu Email xuhy@pumc.edu.cn; yu-jch@163.comIntroduction: Elemene (C15H24) is a sesquiterpene compound extracted from the rhizome of Curcuma herbs. In the past decades, the anti-tumor activity of elemene has been observed in vitro and in some clinical practices. However, pharmacological mechanisms of elemene are not demonstrated adequately, which may lead to improper clinical applications. This study aimed to investigate the anti-tumor effect of elemene nanoemulsion in the mouse model of triple-negative breast cancer (TNBC) and reveal the underlying mechanisms.Methods: The ESR measurement and quantum mechanics simulation were used to characterize the antioxidant ability of elemene nanoemulsion. The murine breast cancer cell line 4T1 cells were inoculated subcutaneously into the left fourth mammary fat pad of BalB/c mice to establish a TNBC mice model. The H&E staining, immunohistochemical staining, DHE staining and Western blot were employed to evaluate the therapeutic effects of the elemene nanoemulsion on the TNBC mice.Results: It was shown that the elemene nanoemulsion prolonged the survival of the triple-negative breast cancer-bearing mice and inhibited the metastasis to lung and liver while did not induce significant cytotoxicity to the tumor cells. Mechanistic studies demonstrated that the elemene nanoemulsion effectively scavenged the reactive oxygen species (ROS) in vitro and in vivo, which decreased the stabilization of hypoxia-inducible factor-1α (HIF-1α) and consequently reduced angiogenesis in the tumor microenvironment as well as decreased the level of NLRP3 inflammasomes and IL-1β production. In addition, the elemene nanoemulsion downregulated the level of IL-1β in the RAW264.7 cells in exposure with LPS.Conclusion: In conclusion, due to the ROS scavenging ability, elemene nanoemulsion effectively inhibited the metastasis of the breast cancer cells to lung and liver and consequently prolonged the survival of TNBC mice.Keywords: elemene, breast cancer, ROS, macrophages, tumor microenvironment

Qiuping Li, Xi Chen, Binru Han Department of Nursing, Xuanwu Hospital, Capital Medical University, Beijing, People’s Republic of ChinaCorrespondence: Binru HanXuanwu Hospital, Capital Medical University, Chang Chun St.45, Xicheng District, Beijing, 100053, People’s Republic of ChinaTel +86 10-83198219Fax +86 10-83198258Email kinv246@126.comBackground: Hemoglobin concentration differs by sex, possibly affecting any association between hemoglobin and frailty. This study aimed to evaluate the potential interaction effect of hemoglobin and sex on frailty in Chinese older inpatients.Methods: A cross-sectional study was conducted between February 2015 and November 2017 in a tertiary hospital. Frailty was defined by the Fried phenotype. Hemoglobin concentration was measured with a standard procedure. Covariates included demographics, clinical characteristics, and serum biomarkers. Logistic regression was applied to examine the association between hemoglobin concentration and frailty. The relative excess risk due to interaction (RERI), attributable proportion due to interaction (AP), and synergy index (SI) were used to evaluate the additive interaction.Results: A total of 619 older inpatients [mean age 69.26± 7.44 years; 334 men, 285 women] were included. The mean hemoglobin concentration was significantly lower in the elderly who were frail (11.9 g/L in frail versus 13.1g/L in non-frail; p< 0.001). In the multivariable regression models, lower hemoglobin in patients was significantly associated with frailty (adjusted odds ratio (OR) = 2.51, 95% CI:1.37, 4.60). The stratified analyses indicated that lower hemoglobin was associated with frailty among older inpatients with different characteristics. Female inpatients with lower hemoglobin had the highest risk of frailty (adjusted OR=6.43, 95%: 2.38, 17.3); there were interactions between hemoglobin and sex on the development of frailty (RERI=4.30, 95% CI=− 1.41, 10.01; AP=0.67, 95% CI=0.37, 0.97;SI=4.80, 95% CI=1.22, 18.84).Conclusions and Implications: Our study provided evidence that sex and lower hemoglobin have an interaction effect on frailty; it is suggested that clinicians may consider sex-specific strategies for the elderly to conform the concept of precision medicine.Keywords: hemoglobin concentration, frailty phenotype, older inpatient, interaction effect

Yanfen Cui, Baoai Han, He Zhang, Hui Liu, Fei Zhang, Ruifang Niu Public Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Breast Cancer Prevention and Therapy, Ministry of Education, Tianjin, 300060, People’s Republic of ChinaCorrespondence: Ruifang Niu; Fei Zhang Tel +86 13752668992; +86 18622221365Email rniu@tmu.edu.cn; feizhang03@tmu.edu.cnBackground and Aim: Uncontrolled proliferation is the most prominent biological feature of tumors. In order to rapidly proliferate, tumor cells regulate their metabolic behavior by controlling the expression of metabolism-related genes (MRGs) to maximize the utilization of available nutrients. In this study, we aimed to construct prognosis models for colorectal adenocarcinoma (COAD) and rectum adenocarcinoma (READ) using MRGs to predict the prognoses of patients.Methods: We first acquired the gene expression profiles of COAD and READ from the TCGA database, and then utilized univariate Cox analysis, Lasso regression, and multivariable Cox analysis to identify the MRGs for risk models.Results: Eight genes (CPT1C, PLCB2, PLA2G2D, GAMT, ENPP2, PIP4K2B, GPX3, and GSR) in the colon cancer risk model and six genes (TDO2, PKLR, GAMT, EARS2, ACO1, and WAS) in the rectal cancer risk model were identified successfully. Multivariate Cox analysis indicated that these two models could accurately and independently predict overall survival (OS) for patients with COAD or READ. Furthermore, functional enrichment analysis was used to identify the metabolism pathway of MRGs in the risk models and analyzed these genes comprehensively. Then, we verified the prognosis model in independent COAD cohorts (GSE17538) and detected the correlations of the protein expression levels of GSR and ENPP2 with prognosis for COAD or READ.Conclusion: In this study, 14 MRGs were identified as potential prognostic biomarkers and therapeutic targets for colorectal cancer.Keywords: metabolism-related gene, colon adenocarcinoma, rectum adenocarcinoma, prognosis, ENPP2, GSR

Shuai Liu,1 Pan Pan,2 Bing Han,3 Jingnan Wang,1 Meili Sun,1 Yuping Sun1 1Department of Oncology, Jinan Central Hospital Affiliated to Shandong University; Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, 250013, Shandong, People’s Republic of China; 2Department of Pathology, Jinan Central Hospital Affiliated to Shandong University; Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, 250013, Shandong, People’s Republic of China; 3Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan, 250012, Shandong, People’s Republic of ChinaCorrespondence: Yuping Sun Tel +86-13370582181Email 13370582181@163.comAbstract: Gastrointestinal stromal tumors (GISTs) are the dominant mesenchymal tumors of the digestive tract. Extragastrointestinal stromal tumors (EGISTs) usually originate outside the gastrointestinal tract without connection to the gastric or intestinal wall. However, EGISTs arising from the vaginal wall are very rare. Here, we report a case of EGIST that occurred in the vagina of a 60-year-old woman. The tumor was present in the posterior vaginal wall. It was surgically excised, and histological examination revealed spindle cell morphology with up to 14 mitoses per 50 high power field (HPF) and necrosis with the tumor-negative margins. Immunohistochemical analyses showed strongly positive CD34, CD117, and DOG-1 expression, but negative SMA, S-100, CD10, desmin, and actin expression. The patient underwent surgery and is currently being followed up. A literature review of EGSTs and treatments is also discussed in this report.Keywords: extragastrointestinal stromal tumors, vagina, c-kit, mutation

Qianhan Lin,1,2,* Xiaoxue Ma,1,2,* Shunxue Hu,3 Rui Li,1,2 Xuan Wei,1,2 Bing Han,1 Yanhui Ma,1 Peishu Liu,1,2,4 Yingxin Pang1,2,4 1Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of China; 2Key Laboratory of Gynecologic Oncology of Shandong Province, Jinan, 250012, Shandong, People’s Republic of China; 3Department of Pathology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of China; 4Shandong Engineering Laboratory for Urogynecology, Jinan, 250012, Shandong, People’s Republic of China*These authors contributed equally to this workCorrespondence: Peishu LiuDepartment of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of ChinaTel +8618560081988Email peishuliu@126.comYingxin PangDepartment of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of ChinaTel +8618560081127Email yingxinpang@126.comPurpose: Nucleolin (NCL) is a multifunctional protein with oncogenic properties. NCL expression levels have been linked to the outcomes of various malignancies, but the clinical value of NCL in patients with endometrial carcinoma (EC) remains unclear. Here, the expression of NCL in EC tissues and its associations with patient outcomes were assessed.Patients and Methods: Data on NCL mRNA expression in EC and adjacent nonneoplastic tissues from The Cancer Genome Atlas (TCGA) were analyzed. In addition, NCL protein expression in 82 endometroid endometrial adenocarcinoma tissues and 15 non-malignant tissues was detected by immunohistochemistry.Results: Elevated NCL expression was markedly correlated with serous endometrial carcinoma (P< 0.001), advanced stage (P=0.029), and grade 3 (P< 0.001). High NCL levels were associated with poorer overall survival (OS) and disease-free survival (DFS) compared with intermediate or low NCL levels (OS: P=0.001, DFS: P=0.006). The multivariate Cox proportional hazards model showed that NCL expression was an independent poor prognostic factor for DFS (HR=1.282, CI=1.027– 1.601, P=0.028). A similar correlation between high expression levels of NCL and unfavorable DFS was found in endometrioid endometrial adenocarcinoma (HR=1.411, CI=1.083– 1.840, P=0.011). Positive extra-nuclear NCL expression (HR=3.377, 95% CI=1.029– 11.186, P=0.046) and low nuclear NCL expression (HR=0.233, 95% CI=0.068– 0.796, P=0.020) were independent prognostic factors for DFS in endometrioid endometrial adenocarcinoma.Conclusion: Heterotopic NCL is a potential prognostic biomarker for EC. Inhibiting the distribution of NCL from the nucleus to the cytoplasm and membrane may be a promising therapeutic strategy to improve outcomes in patients with EC with high NCL expression.Keywords: nucleolin, endometrial carcinoma, prognostic marker, TCGA

Lei Wang,1 Wei Cai,2 Bing Han,1 Jue Zhang,1 Bing Yu,1 Ming Chen1 1Burn and Plastic Surgery, Zhongda Hospital Affiliated Southeast University, Nanjing, 210009, People’s Republic of China; 2Burn and Plastic Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, People’s Republic of ChinaCorrespondence: Wei CaiBurn and Plastic Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210009, People’s Republic of ChinaEmail caiwei8656@163.comBackground: Malignant melanoma was characterized by insensitive chemotherapy, drug resistance, and high metastatic ability, which resulted in the main reason for the mortality among skin-related cancers. The current agents were not sufficient to improve the treatment status of melanoma patients, and it was still needed to develop new chemotherapeutic drugs for melanoma. Our study aimed to study the anticancer effects and potential mechanisms of ouabain on melanoma cells.Methods: The inhibitory effects of ouabain were determined by CCK8 and colony formation assays, and the morphological changes of melanoma cells were observed by inverted microscope. The apoptosis induction and cell cycle distribution were detected by annexin V/PI double staining and PI staining, respectively. The expression of the biomarker proteins in apoptosis and G2/M phase were determined by Western blotting analysis. The effects of ouabain on the migration of melanoma cells were measured by transwell migration assay and wound closure analysis. The potential mechanisms of ouabain in melanoma cells were analyzed by transcriptome sequencing.Results: Our present study demonstrated that ouabain exhibited strong inhibitory effects on cell proliferation and triggered dramatical morphological changes of melanoma cells. Moreover, ouabain induced significant apoptosis in A375 rather than SK-Mel-28 cells via upregulation of bax expression and downregulation of bcl-2 expression. Consistently, ouabain treatment induced cell cycle arrest at G2/M phase in both A375 and SK-Mel-28 cells via upregulation of cyclin B1 and downregulation of cdc2 and cdc25c. Importantly, ouabain suppressed the migration of A375 and SK-Mel-28 cells. Furthermore, the transcriptome sequencing demonstrated that p53 and MAPK signaling pathway might play important roles in the inhibitory effects of ouabain.Conclusion: Our study revealed that ouabain exhibited dramatical anticancer effects, which provided a novel application for cardiac glycoside drugs in the clinical treatment of melanoma.Keywords: melanoma, ouabain, apoptosis, cell cycle, migration

Cuiping Liu,1 Yizhou Jiang,2 Baosan Han3 1Department of Breast Surgery, Affiliated Hospital of Chifeng University, Chifeng, Inner Mongolia 024005, People’s Republic of China; 2Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, People’s Republic of China; 3Department of Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, People’s Republic of ChinaCorrespondence: Baosan HanXin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, No. 1665 Kongjiang Road, Yangpu District, Shanghai 200092, People’s Republic of ChinaTel +86-2165792951Email shhanbs311@126.comIntroduction: Triple-negative breast cancer (TNBC) is the most aggressive malignancy of breast cancer, which represents about 20% of all cases. The prognosis of TNBC remains unfavorable due to the lack of targeted therapy and chemoresistance. The aim of this study is to investigate the role of miR-613 in TNBC.Material and Methods: Quantitative RT-PCT was used to explore the expression of miR-613 in breast cancer clinical samples and cell lines. MTT, colony formation assay, spheroid formation assay and xenograft tumor growth assay were used to investigate the role of miR-613 in vitro and in vivo. Cell apoptosis and surface marker expression were measured by flow cytometry. Dual-luciferase reporter assay was used to explore the function of miR-613 in regulating FAM83A 3ʹUTR. Immunohistochemical staining was used to investigate the expression of FAM83A in TNBC tissues.Results: We found that miR-613 expression was significantly downregulated in breast cancer tissues and was even lower in TNBC compared with that in other types of breast cancer. A similar result was found in breast cancer cell lines. Further analysis indicated that miR-613 could suppress TNBC cell growth, chemoresistance and stem-cell-like phenotype. Moreover, we also demonstrated that miR-613 suppressed tumorigenesis in vivo. Mechanically, we explored the downstream target of miR-613 and identified that miR-613 could directly bind to the 3ʹUTR of FAM83A, which contributed to the miR-613 mediated tumor suppression. The expression of miR-613 and FAM83A was negatively correlated. Restoring the expression of FAM83A attributed to the chemoresistance and stemness of TNBC cells.Conclusion: We demonstrated that loss of miR-613 was critical for TNBC malignancy and restoring its expression could be served as a potential approach for TNBC treatment.Keywords: triple-negative breast cancer, miR-613, FAM83A, chemoresistance, stemness

Dong Liu,1– 3 Yanjun Chen,1,2 Fang Li,1,2 Cunwu Chen,1,2 Peipei Wei,1,2 Deli Xiao,4,5 Bangxin Han1,2 1School of Biological and Pharmaceutical Engineering, West Anhui University, Lu’an 237012, People’s Republic of China; 2Anhui Engineering Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources, Lu’an 237012, People’s Republic of China; 3School of Biomedical Sciences and Medical Engineering, Southeast University, Nanjing 210009, People’s Republic of China; 4Department of Analytical Chemistry, China Pharmaceutical University, Nanjing 210009, People’s Republic of China; 5Key Laboratory of Biomedical Functional Materials, China Pharmaceutical University, Nanjing 210009, People’s Republic of ChinaCorrespondence: Deli XiaoDepartment of Analytical Chemistry, China Pharmaceutical University, Nanjing 210009, People’s Republic of ChinaTel +86 25 86185160Email xiao49562001@163.comBangxin HanSchool of Biological and Pharmaceutical Engineering, West Anhui University, Lu’an 237012, People’s Republic of ChinaTel +86 564 3305073Email hanbx1978@sina.comPurpose: Pulmonary surfactants (eg, sinapultide) are widely used for the treatment of lung injury diseases; however, they generally induce poor therapeutic efficacy in clinics. In this study, sinapultide-loaded microbubbles (MBs) were prepared and combined with ultrasound (US) treatment as a new strategy for improved treatment of lung injury diseases.Methods: The combination treatment strategy of MBs combined with ultrasound was tested in a lipopolysaccharide (LPS)-induced mouse model of alveolar epithelial cells (AT II) and acute lung injury. Firstly, cytotoxicity, cytokines, and protein levels in LPS-mediated AT II cells were assessed. Secondly, the pathological morphology of lung tissue, the wet/dry (W/D) weight ratio, cytokines, and protein levels in LPS-mediated acute lung injury mice after treatment with the MBs were evaluated. Moreover, histology examination of the heart, liver, spleen, lung and kidney of mice treated with the MBs was performed to initially evaluate the safety of the sinapultide-loaded MBs.Results: Sinapultide-loaded MBs in combination with ultrasound treatment significantly reduced the secretion of inflammatory cytokines and increased the expression of surfactant protein A (SP-A) in AT II cells. Furthermore, the pathological morphology of lung tissue, the wet/dry (W/D) weight ratio, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and SP-A expression level of mice treated with MBs and ultrasound were significantly improved compared to those of non-treated mice. In addition, the histology of the examined organs showed that the MBs had a good safety profile.Conclusion: Sinapultide-loaded MBs combined with ultrasonic treatment may be a new therapeutic option for lung injury diseases in the clinic.Keywords: sinapultide, microbubbles, lipopolysaccharide, lung injury, safety

Yali Du,1 Yuzhou Huang,1 Wenzhe Zhou,1 Xinjian Liu,2 Fangfei Chen,1 Chen Yang,1 Miao Chen,1 Jing Ruan,1 Bing Han1 1Department of Hematology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, People’s Republic of China; 2Department of Hematology, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Institute of Hematology of Henan Province, Zhengzhou, People’s Republic of ChinaCorrespondence: Bing HanDepartment of Hematology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, People’s Republic of ChinaEmail hanbing_li@sina.com.cnBackground: For symptomatic non-severe aplastic anemia (NSAA) patients who cannot afford anti-thymocyte globulin (ATG) or allogeneic hematopoietic stem cell transplantation (HSCT), tacrolimus (FK) may be an option if these patients do not respond or become tolerant to cyclosporine A (CsA).Methods: We enrolled 101 NSAA patients who were refractory or intolerant to CsA with no chance of HSCT or ATG treatment and treated these patients with tacrolimus for at least 6 months, with follow-up for at least one year.Results: The overall response rate (ORR) was 38.6% (complete response: 9.9%; partial response: 28.7%), and the median time to optimal response was 6 (3～10) months. Thirty-two (31.7%) cases had elevated creatinine levels. Eight (7.9%) cases had elevations in AST/ALT. A total of 25.6% (10/39) of patients relapsed at the end of follow-up. Age (P=0.0005), FK concentration (4.0∼ 12 ng/mL, P=0.0005) and intolerance to CsA (P=0.012) were the independent risk factors for ORR. Treg cell levels pre-FK treatment were much lower than those of healthy controls (3.7± 0.6% vs 6.8± 0.7%, P=0.0004) but increased significantly after FK treatment (3.7± 0.6% vs 7.1± 0.8%, P=0.0039).Conclusion: Our data suggest that tacrolimus is a salvage treatment for patients with NSAA that is refractory or intolerant to CsA.Keywords: non-severe aplastic anemia, tacrolimus, cyclosporine A

Rong Qiao,1,* Feifei Di,2,* Jun Wang,2 Yujie Wei,2 Yanman Zhang,2 Tian Xu,3 Yue Wang,1 Wanjian Gu,3 Baohui Han,1 Rongxi Yang2,4 1Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai 200030, People’s Republic of China; 2Department of Research and Academic, Nanjing TANTICA Biotechnology Co. Ltd, Nanjing 210000, People’s Republic of China; 3Department of Clinical Laboratory, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210000, People’s Republic of China; 4Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 210000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Rongxi Yang; Baohui Han Email rongxiyang@njmu.edu.cn; 18930858216@163.comBackground: Early detection is essential to improve the survival and life quality of lung cancer (LC) patients. Changes of peripheral blood DNA methylation could be associated with malignancy but were mostly studied in Caucasians.Methods: Here, in a Chinese population, we performed mass spectrometry assays to investigate the association between very early stage LC and methylation levels of RAPSN in the peripheral blood by a case–control cohort using of 221 LC patients (93.2% LC at stage I) and 285 unrelated cancer free control individuals.Results: The odds ratios (ORs) of all CpG sites were evaluated for their risk to LC using inter-quartile analyses by logistic regression. In general, we observed an association between very early LC and decreased methylation of RAPSN_CpG_1.15 and RAPSN_CpG_3.4 (referring to Q4, OR range from 1.64 to 1.81, p< 0.05). Stratified by gender, while hypomethylation of RAPSN_CpG_1.15, RAPSN_CpG_3.4 and RAPSN_CpG_7.14 were associated with LC in males (referring to Q4, ORs range from 1.94 to 2.31, p< 0.05), RAPSN_CpG_2 and RAPSN_CpG_5 showed significantly lower methylation in female LC patients comparing to controls (referring to Q4, ORs range from 2.49 to 3.60, p< 0.05). The risk of RAPSN hypomethylation to LC was enhanced by aging, and typically for people older than 55 years (referring to Q4, ORs range from 2.17 to 3.61 in six out of all 10 analyzed CpG groups, p< 0.05).Conclusion: Our study reveals an association between RAPSN hypomethylation in peripheral blood and LC and suggests the occurrence of altered blood-based methylation at the early stage of cancer.Keywords: LC, early detection, DNA methylation, RAPSN, peripheral blood

Leishmaniasis is an infectious disease caused by protozoan parasites belonging to the genus Leishmania for which there are no approved human vaccines. Infections localise to different tissues in a species-specific manner with the visceral form of the disease caused by Leishmania donovani and L. infantum being the most deadly in humans. Although Leishmania spp. parasites are predominantly intracellular, the visceral disease can be prevented in dogs by vaccinating with a complex mixture of secreted products from cultures of L. infantum promastigotes. With the logic that extracellular parasite proteins make good subunit vaccine candidates because they are directly accessible to vaccine-elicited host antibodies, here we attempt to discover proteins that are essential for in vitro growth and host infection with the goal of identifying subunit vaccine candidates. Using an in silico analysis of the Leishmania donovani genome, we identified 92 genes encoding proteins that are predicted to be secreted or externally anchored to the parasite membrane by a single transmembrane region or a GPI anchor. By selecting a transgenic L. donovani parasite that expresses both luciferase and the Cas9 nuclease, we systematically attempted to target all 92 genes by CRISPR genome editing and identified four that were required for in vitro growth. For fifty-five genes, we infected cohorts of mice with each mutant parasite and by longitudinally quantifying parasitaemia with bioluminescent imaging, showed that nine genes had evidence of an attenuated infection although all ultimately established an infection. Finally, we expressed two genes as full-length soluble recombinant proteins and tested them as subunit vaccine candidates in a murine preclinical infection model. Both proteins elicited significant levels of protection against the uncontrolled development of a splenic infection warranting further investigation as subunit vaccine candidates against this deadly infectious tropical disease.

Bin Han1 ,* Zijie Pei2 ,* Lei Shi,3 Qian Wang,4 Chen Li,1 Boyuan Zhang,1 Xuan Su,1 Ning Zhang,1 Lixiao Zhou,1 Bo Zhao,5 Yujie Niu,3,6 Rong Zhang1,6 1Department of Toxicology, Hebei Medical University, Shijiazhuang 050017, Hebei, People’s Republic of China; 2Department of Pathology, Medical School, China Three Gorge University, Yichang 443002, People’s Republic of China; 3Occupational Health and Environmental Health, Hebei Medical University, Shijiazhuang 050017, Hebei, People’s Republic of China; 4Experimental Center, Hebei Medical University, Shijiazhuang 050017, Hebei, People’s Republic of China; 5Department of Laboratory Diagnosis, Hebei Medical University, Shijiazhuang 050017, Hebei, People’s Republic of China; 6Hebei Key Laboratory of Environment and Human Health, Hebei Medical University, Shijiazhuang 050017, Hebei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Rong ZhangDepartment of Toxicology, Hebei Medical University, 361 Zhongshan East Road, Shijiazhuang, Hebei 050017, People’s Republic of ChinaFax +86-311-86265605Email rongzhang@hebmu.edu.cnIntroduction: Because of the increased production and application of manufactured Nano-TiO2 in the past several years, it is important to investigate its potential hazards. TiO2 is classified by IARC as a possible human carcinogen; however, the potential mechanism of carcinogenesis has not been studied clearly. The present study aimed to investigate the mechanism of DNA damage in rat lung and extra-pulmonary organs caused by TiO2nanoparticles.Methods: In the present study, SD rats were exposed to Nano-TiO2 by intratracheal injection at a dose of 0, 0.2, or 1 g/kg body weight. The titanium levels in tissues were detected by ICP-MS. Western blot was used to detect the protein expression levels. The DNA damage and oxidative stress were detected by comet assay and ROS, MDA, SOD, and GSH-Px levels, respectively.Results: The titanium levels of the 1 g/kg group on day-3 and day-7 were significantly increased in liver and kidney as well as significantly decreased in lung compared to day-1. ROS and MDA levels were statistically increased, whereas SOD and GSH-Px levels were statistically decreased in tissues of rats in dose-dependent manners after Nano-TiO2 treatment. PI3K, p-AKT/AKT, and p-FOXO3a/FOXO3a in lung, liver, and kidney activated in dose-dependent manners. The levels of DNA damage in liver, kidney, and lung in each Nano-TiO2 treatment group were significantly increased and could not recover within 7 days. GADD45α, ChK2, and XRCC1 in liver, kidney, and lung of rats exposed to Nano-TiO2 statistically increased, which triggered DNA repair.Conclusion: This work demonstrated that Ti could deposit in lung and enter extra-pulmonary organs of rats and cause oxidative stress, then trigger DNA damage through activating the PI3K-AKT-FOXO3a pathway and then promoting GADD45α, ChK2, and XRCC1 to process the DNA repair.Keywords: Nano-TiO2, DNA damage, PI3K/AKT/FOXO3a signaling pathway, DNA repair, GADD45α/ChK2/XRCC1 signaling pathway

Bin Han,1 E Shaolong,1 Lan Luan,2 Nanyang Li,2 Xuefeng Liu1 1Department of Urology, Shengjing Hospital of China Medical University, Shenyang, People’s Republic of China; 2Department of Pathology, Central Hospital Affiliated to Shenyang Medical College, Shenyang, People’s Republic of ChinaCorrespondence: Xuefeng LiuDepartment of Urology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, People’s Republic of ChinaEmail xuefengliu78@163.comIntroduction: Accumulating evidence has demonstrated that circular RNAs (circRNAs) play a key role in the tumorigenesis of various types of cancers, including clear cell renal cell carcinoma (ccRCC).Materials and Methods: Reverse transcription-quantitative polymerase chain reaction was used to detect the expression of circRNA homeodomain interacting protein kinase 3 (circHIPK3) and microRNAs (miRNAs), including miR-508-3p. The clinical measurement of circHIPK3 was evaluated by Kaplan–Meier survival analysis and receiver operating characteristic analysis. Cell Counting Kit-8 and Transwell chamber assays were performed to determine the changes in the proliferative and metastatic ability of A498 and 786-O cells. C-X-C motif chemokine ligand 13 (CXCL13) protein expression was detected by Western blot analysis. The targeted binding effect between miR-508-3p and circHIPK3 or CXCL13 was confirmed by constructed luciferase and RNA immunoprecipitation (RIP) assays, respectively. Fluorescence in situ hybridization (FISH) assay was used to measure the subcellular localization of circHIPK3 and miR-508-3p.Results: It was found that circHIPK3 was markedly upregulated in ccRCC tissue and cell lines, and circHIPK3-upregulation was closely correlated with poor clinicopathological features in patients with ccRCC. It was found that both miR-508-3p and circHIPK3 were localized in the cytoplasm of ccRCC cells. The up- and downregulation of circHIPK3 positively regulated ccRCC cell proliferation and metastasis, and this regulatory effect was reversed by miR-508-3p. Through luciferase and RIP assays, it was confirmed that circHIPK3 could interacted with miR-508-3p. Furthermore, it was revealed that CXCL13, which was negatively correlated with miR-508-3p, was upregulated in ccRCC. It was also shown that CXCL13 was a downstream target of miR-508-3p. miR-508-3p suppressed ccRCC cell proliferation and metastasis by targeting CXCL13. Lastly, it was demonstrated that circHIPK3 promoted CXCL13 to facilitate ccRCC cell proliferation and metastasis by decoying miR-508-3p.Conclusion: In brief, the results of the present study showed that circHIPK3 promoted ccRCC cell proliferation and metastasis by altering miR-5083p/CXCL13 signaling. The present findings might provide a novel target for the molecular treatment of ccRCC.Keywords: circHIPK3, miR-508-3p, CXCL13, proliferation, metastasis, ccRCC

Hai Zhang,1,* Xiaohua Yang,2,* Fang Hu,1,* Changhui Li,1 Jianlin Xu,1 Wei Nie,1 Yinchen Shen,1 Yuqing Lou,1 Baohui Han,1 Hua Zhong,1 Xueyan Zhang1 1Department of Pulmonary, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, People’s Republic of China; 2Central Laboratory, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xueyan Zhang; Hua ZhongDepartment of Pulmonary, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, People’s Republic of ChinaTel +86 18017321319; +86 13818200560Fax +86-021-62821990Email zxychest@163.com; eddiedong8@hotmail.comBackground and Objective: The first-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have shown significant therapeutic effects on patients harboring sensitive EGFR mutations, while the mechanisms related to drug resistance have still remained elusive. This study aimed to indicate the relationship between the expression level of Wnt5a with therapeutic effects of first-generation EGFR-TKIs on lung adenocarcinoma patients harboring sensitive EGFR mutations.Methods: The medical records of 75 lung adenocarcinoma patients harboring sensitive EGFR mutations, who were admitted to our hospital and received first-generation EGFR-TKIs from June 1, 2010 to December 31, 2016, were analyzed. According to the efficacy of first-generation EGFR-TKIs, patients were divided into ineffective groups (progression-free survival (PFS) < 5 months) and effective groups (PFS > 26 months). Immunofluorescence staining, immunohistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) methods were utilized to detect the expression level of Wnt5a in the two groups.Results: Among 75 patients, 36 patients were sensitive to first-generation EGFR-TKIs (effective group) and 39 patients were resistant to first-generation EGFR-TKIs (ineffective group). The location of Wnt5a was detected by immunofluorescence staining. Immunohistochemical staining demonstrated that the expression level of Wnt5a in the ineffective group was significantly higher than that in the effective group (P=0.0216). Besides, results of RT-PCR showed that the relative expression level of Wnt5a was remarkably higher in the ineffective group than that in the effective group (P=0.0135).Conclusion: The expression level of Wnt5a was found to be associated with therapeutic effects of first-generation EGFR-TKIs in lung adenocarcinoma patients harboring sensitive EGFR mutations.Keywords: tyrosine-kinase inhibitors, Wnt5a, epidermal growth factor receptor mutation, therapeutic effect

Jie Qian,1,2,* Xueyan Zhang,2,* Bo Zhang,2 Bo Yan,2 Lin Wang,3 Ping Gu,2 Weimin Wang,2 Huimin Wang,2 Baohui Han2 1Department of Emergency Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 2Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China; 3Department of Laboratory Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Baohui HanDepartment of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, 241 # West Huaihai Road, Shanghai 230030, People’s Republic of ChinaTel +86 21 62821990 ext.1421Fax +86 21 32260806Email 18930858216@163.comPurpose: Hepatic injury is a common side effect following tyrosine kinase inhibitor (TKI) therapy and our understanding usually comes from clinical trials. In this retrospective study, we aimed to investigate the characteristics, risk factors and regimen-related differences of epidermal growth factor receptor (EGFR)-TKI-related hepatic toxicity in patients with advanced lung adenocarcinoma (LAD).Patients and Methods: Liver function tests were documented in 424 patients admitted into the Shanghai Chest Hospital between January 2014 and December 2016 with advanced (IIIB/IV) LAD who received first-line gefitinib, erlotinib or icotinib. Hepatotoxicity was graded according to the Common Terminology Criteria for Adverse Events (CTCAE), version 4.0. The clinical spectrum and onset time of hepatic injury were evaluated. The risk factors of hepatic dysfunction were determined using a logistic regression analysis.Results: A total of 87 (20.5%) patients experienced hepatotoxicity and 5.7% were of grade 3/4 liver dysfunction. The median onset time of hepatotoxicity was 7 weeks. Presence of hepatitis virus (HR: 2.593, 95% CI: 1.090– 6.170, P=0.031) and pretreatment liver impairment (HR: 3.460, 95% CI: 1.746– 6.855, P< 0.001) were risk factors associated with increased risk of hepatotoxicity. Gefitinib (HR: 1.872, 95% CI: 1.028– 3.412, P=0.040) and erlotinib (HR: 3.578, 95% CI: 1.683– 7.609, P=0.001) had increased risk of hepatotoxicity compared to icotinib.Conclusion: The different toxic profile of EGFR-TKIs should be taken into account in the choice of treatment based on the patients’ comorbidity.Keywords: lung adenocarcinoma, hepatotoxicity, tyrosine kinase inhibitor, gefitinib, erlotinib, icotinib

Wen-Jing Zhu,1 Ying Yan,2 Jiu-Wei Zhang,1 Yan-Dong Tang,3 Bo Han4 1Abdominal Ultrasonic Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, People’s Republic of China; 2Oncology Department, The First Hospital of Harbin, Harbin, Heilongjiang Province, People’s Republic of China; 3Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang Province, People’s Republic of China; 4Oncology Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, People’s Republic of ChinaCorrespondence: Bo HanOncology Department, The First Affiliated Hospital of Harbin Medical University, 23 Post Street, Harbin 150001, Heilongjiang Province, People’s Republic of ChinaEmail hanbo1964@163.comAim: This study aimed to investigate the effect and mechanism of miR-26a-5p on proliferation and apoptosis of hepatocellular carcinoma (HCC) cells.Methods: RT-PCR was used to analyze the expression of miR-26a-5p in HCC cells and its targeted gene HMGA2 mRNA determined by biological information prediction. The rate of proliferation, invasion, apoptosis, and expression levels of related proteins of HCC cells overexpressing miR-26a-5p or those after knocking down HMGA2 expression were detected by MTT, invasion and apoptosis rate tests. Moreover, the apoptosis-promoting protein bax was upregulated and the anti-apoptosis-related protein Bcl-2 was downregulated.Results: RT-qPCR results showed that the level of miR-26a-5p was downregulated in HCC tissues and cells, and the expression of HMGA2 was upregulated; besides, the expression of miR-26a-5p and HMGA2 was negatively correlated; miR-26a-5p was correlated with tumor diameter, differentiation degree, TNM staging and lymph node metastasis. Cell tests confirmed that miR-26a-5p functioned in tumor suppression, including inhibiting cell proliferation and invasion in two hepatocellular carcinoma cell lines and promoting apoptosis. Bioinformatics prediction and subsequent experiments proved that HMGA2 was the direct target of miR-26a-5p; moreover, after knocking down HMGA2 expression in HCC cells, cell proliferation and invasion ability were significantly inhibited, and apoptosis rate increased significantly.Conclusion: miR-26a-5p can inhibit the proliferation and invasion of HCC cells and promote their apoptosis by directly targeting HMGA2. Abnormal decrease of miR-26a-5p and increase of its target HMGA2 are important factors that may participate in the occurrence and development of HCC. miR-26a-5p may be a new potential target for its treatment.Keywords: miR-26a-5p, hepatocellular carcinoma cells, proliferation, apoptosis, mechanism research

Bing Han,1,* Yue Yang,1,2,* Jinglin Chen,1 Huan Tang,1 Yuxin Sun,1 Zheng Zhang,1 Zeng Wang,1 Yan Li,1 Yao Li,1 Xue Luan,1 Qianwen Li,1 Zhihui Ren,1 Xiaowei Zhou,1 Dengli Cong,1 Zhiyi Liu,1 Qin Meng,1 Fei Sun,1 Jin Pei1 1Department of Biopharmacy, School of Pharmaceutical Sciences, Jilin University, ChangChun, People’s Republic of China; 2Department of Pharmacy, Ministry of Health Service, Chinese PLA General Hospital, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jin Pei Tel +86 431 85619725Email peijin@jlu.edu.cnBackground: Breast cancer is the leading cause of cancer death in women. Chemotherapy to inhibit the proliferation of cancer cells is considered to be the most important therapeutic strategy. The development of long-circulating PEG and targeting liposomes is a major advance in drug delivery. However, the techniques used in liposome preparation mainly involve conventional liposomes, which have a short half-life, high concentrations in the liver and spleen reticuloendothelial system, and no active targeting.Methods: Four kinds of paclitaxel liposomes were prepared and characterized by various analytical techniques. The long-term targeting effect of liposomes was verified by fluorescence detection methods in vivo and in vitro. Pharmacokinetic and acute toxicity tests were conducted in ICR mice to evaluate the safety of different paclitaxel preparations. The antitumor activity of ES-SSL-PTX was investigated in detail using in vitro and in vivo human breast cancer MCF-7 cell models.Results: ER-targeting liposomes had a particle size of 137.93± 1.22 nm and an acceptable encapsulation efficiency of 88.07± 1.25%. The liposome preparation is best stored at 4°C, and is stable for up to 48 hrs. Cytotoxicity test on MCF-7 cells demonstrated the stronger cytotoxic activity of liposomes in comparison to free paclitaxel. We used the near-infrared fluorescence imaging technique to confirm that ES-SSL-PTX was effectively targeted and could quickly and specifically identify the tumor site. Pharmacokinetics and acute toxicity in vivo experiments were carried out. The results showed that ES-SSL-PTX could significantly prolong the half-life of the drug, increase its circulation time in vivo, improve its bioavailability and reduce its toxicity and side effects. ES-SSL-PTX can significantly improve the pharmacokinetic properties of paclitaxel, avoid allergic reaction of the original solvent, increase antitumor efficacy and reduce drug toxicity and side effects.Conclusion: ES-SSL-PTX has great potential for improving the treatment of breast cancer, thereby improving patient prognosis and quality of life.Keywords: breast cancer, estrogen receptors, paclitaxel, long-acting liposomes, targeted drug delivery

Zhenhua Cui,1 Bingbing Han,2 Xianren Wang,1 Zhiwen Li,1 Jianxin Wang,1 Yanfeng Lv1 1Department of Colorectal & Anal Surgery, The Second Hospital of Shandong University, Jinan 250033, People’s Republic of China; 2Microcirculation Laboratory, Shandong University of Traditional Chinese Medicine, Jinan, 250014, People’s Republic of ChinaCorrespondence: Yanfeng LvDepartment of Colorectal & Anal Surgery, The Second Hospital of Shandong University, No. 247 BeiYuan Street, Jinan 250033, People’s Republic of ChinaEmail ttdpilsxpsle11@163.comIntroduction: Long non-coding RNAs (lncRNAs) have obtained increasing attention due to their regulatory functions in many cancers. This work aimed to investigate the functional roles of lncRNA TTN-AS1 in colorectal cancer (CRC) and to explore the underlying mechanisms.Methods: The expression profiles of TTN-AS1 and miR-497 in CRC tissues and cell lines were determined by RT-qPCR analysis. MTT assay, transwell assay, western blot analysis, and xenograft tumors in nude mice were employed to analyze the effects of TTN-AS1 on the proliferation, migration, invasion, EMT, and in vivo tumorigenesis of CRC cells. Bioinformatics analysis and dual-luciferase reporter assay determined the direct binding relation between TTN-AS1 and miR-497 in CRC.Results: We observed a significant increase of TTN-AS1 expression level in CRC tissues and cell lines compared with normal counterparts. High expression of TTN-AS1 predicted a poor prognosis and was correlated with aggressive clinicopathological characteristics in CRC patients. Functionally, gain- and loss-of-function studies indicated that TTN-AS1 knockdown suppressed the proliferation, migration, invasion and epithelial–mesenchymal transition of CRC cells in vitro, whereas TTN-AS1 overexpression showed the complete opposite effects. Mechanistically, we found that TTN-AS1 could directly interact with miR-497, and co-transfection with miR-497 mimics blocked the activation of PI3K/Akt/mTOR signaling, and reversed the effects of TTN-AS1 overexpression in CRC cells.Conclusion: To conclude, our findings provide novel insight into CRC tumorigenesis and indicate that TTN-AS1 may serve as a potential therapeutic target for CRC treatment.Keywords: colorectal cancer, long non-coding RNA TTN-AS1, epithelial-mesenchymal transition, microRNA-497, PI3K/Akt/mTOR signaling

Bing Han,1,2,* Jiawei Huang,3,* Zhenjie Yang,1 Jiaqi Zhang,3 Xiaomin Wang,3 Ning Xu,3 Haining Meng,3 Junyu Wu,3 Qiao Huang,3 Xi Yang,3 Ruowu Shen,3 Chuandong Sun1 1Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, Qingdao City 266003, Shandong Province, People’s Republic of China; 2Asian Liver Center, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA; 3Department of Special Medicine, School of Basic Medical College, Qingdao University, Qingdao City 266021, Shandong Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Chuandong SunDepartment of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, No. 16 Jiangsu Road, Qingdao City 266003, Shandong Province, People’s Republic of ChinaTel +86 18661802672Email sunchuandong@hotmail.comRuowu ShenDepartment of Special Medicine, School of Basic Medical College, Qingdao University, Qingdao City 266021, Shandong Province, People’s Republic of ChinaTel +86 18661785993Email shenruowu@aliyun.comPurpose: To explore the effect of miR-449a inhibits migration and invasion by targeting Notch1 and regulating epithelial–mesenchymal transition (EMT) in hepatocellular carcinoma (HCC), and further study on the molecular mechanism.Patients and methods: The expression of miR-449a and Notch1 in HCC cells and tissues was detected by qRT-PCR. The HCC cell line HCCLM3 and SMMC-7721 were transfected with miR-449a. The invasion and migration of HCC cell lines were detected by transwell assay and wound healing assay. The Notch pathway and EMT related protein were detected with Western Blotting. The specific binding site of mir-449a on notch1 gene was detected by luciferase assay.Results: We found the expression of miR-449a was related to short-term recurrence of hepatocellular carcinoma after hepatectomy and acted as independent risk factors of DFS and OS. The expression of miR-449a decreased in tumor tissues and HCC cell lines, but the expression of Notch1 increased. The overexpressed miR-449a promoted the invasiveness in vitro by regulating EMT via Notch pathway. Mechanically, miR-449a inhibited the translation of Notch1 protein by binding to 3ʹ UTR of its mRNA directly.Conclusion: miR-449a is short-term recurrence-related miRNA and inhibits the invasion and metastasis ability of HCC cells by regulating EMT via Notch pathway. miR-449a may be a new effective therapeutic target for HCC.Keywords: signal pathway, microRNA, epithelial-mesenchymal transition, bioinformatics

Mao Zhang,1 Mei-Sze Chua,2 Jie Hu,3 Haoran Li,1 Shun Zhang,1 Liqun Wu,1 Bing Han1 1Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, People’s Republic of China; 2Asian Liver Center, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA; 3Department of General Surgery, The Third Xiangya Hospital of Central South University, Changsha, Hunan, People’s Republic of ChinaCorrespondence: Bing Han; Liqun WuDepartment of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, No.16 Jiangsu Road, Qingdao, Shandong 266003, People’s Republic of ChinaTel +86-186 6180 8308Fax +86-532-8291 1323Email wfkevinhan@163.com; wulq581031@163.comPurpose: In this study, we established the inflammatory factor grade system (IFGs) based on the hepatocellular carcinoma (HCC) microenvironment to investigate the role of inflammatory factor grade (IFG) in predicting the prognosis of patients with American Joint Committee on Cancer (AJCC) stage I-II.Patients and methods: We enrolled 87 HCC patients with AJCC stage I-II who underwent R0 resection between 2000 and 2012 and had paraffin-embedded specimens. Immunohistochemistry (IHC) was performed to investigate the expression of 12 inflammatory factors and then to establish the IFGs (grade A or B) based on the IHC data. Subsequently, Kaplan-Meier and Cox univariate/multivariate survival analyses were performed to examine the potential prognostic significance.Results: Higher IFG (IFG-B) is significantly associated with greater tumor size (P=0.037), and IFG-B predicts a worse disease-free survival (DFS, P

Tao Wang,1 Tao Wen,1 Hongmin Li,2 Bing Han,2 Suisui Hao,1 Chuan Wang,1 Qiang Ma,1 Jie Meng,1 Jian Liu,1 Haiyan Xu11Department of Biomedical Engineering, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 2Department of Hematology, Peking Union Medical College Hospital, Beijing, People’s Republic of ChinaBackground: Chronic myeloid leukemia (CML) is a myeloproliferative disorder due to the existence of BCR-ABL fusion protein that allows the cells to keep proliferating uncontrollably. Although tyrosine kinase inhibitors can inhibit the activity of BCR-ABL fusion protein to trigger the cells apoptosis, drug resistance or intolerance exists in part of CML patients. Arsenic sulfide in its raw form (r-As4S4) can be orally administrated and certain therapeutic effects have been found out in the treatment of hematologic malignancies through inducing cell apoptosis.Methods: In this work, a water-dissolvable arsenic sulfide nanoformualtion (ee-As4S4) composed of As4S4 particulates with 470 nm in diameter and encapsulated by a kind of hydrophilic polymer was fabricated and applied to the CML cell line K562, K562/AO2 and primary cells from the bone marrow of CML patients.Results: Results showed that instead of inhibiting the activity of BCR-ABL, ee-As4S4 induced direct degradation of BCR-ABL in K562 cells within 6 hr incubation, followed by the occurrence of erythroid differentiation in K562 after 72 hr incubation, evidenced by the significantly upregulated CD235a and benzidine staining, which was not detectable with r-As4S4. The ee-As4S4-induced erythroid differentiation was also observed in K562/AO2 cells and bone marrow mononuclear cells of CML patients. Mechanistic studies indicated that ee-As4S4 induced autophagy by downregulating the level of intracellular ROS and hypoxia-inducible factor-1α significantly, which led to the subsequent degradation of BCR-ABL. When the concentration was increased, ee-As4S4 induced much more significant apoptosis and cell cycle arrest than r-As4S4, and the cytotoxicity of the former was about 178 times of the latter.Conclusion: ee-As4S4 was capable of inducing significant erythroid differentiation of CML cells by inducing the direct degradation of BCR-ABL; the new effect could improve hematopoietic function of CML patients as well as inhibit the leukemic cell proliferation.Keywords: chronic myeloid leukemia, erythroid differentiation, As4S4, ROS

Binru Han, Qiuping Li, Xi ChenDepartment of Nursing, Xuanwu Hospital, Capital Medical University, Beijing, People’s Republic of ChinaPurpose: To determine the association between frailty and postoperative complications in elderly Chinese patients and to determine whether addition of frailty assessment improves the predictive ability of the American Society of Anesthesiologists (ASA) score, Physiological and Operative Severity Score for the Enumeration of Mortality and Morbidity (POSSUM), and Estimation of Physiologic Ability and Surgical Stress (E-PASS) score.Patients and methods: A prospective cohort study was conducted in a tertiary hospital. Elderly patients undergoing major thoracic or abdominal surgery were included. Frailty phenotype and ASA, POSSUM, and E-PASS scores were assessed. Demographic, preoperative, and surgical variables were extracted from medical records. Primary outcome measure was in-hospital Clavien-Dindo ≥ grade II complications. Multiple logistic regression was used to examine the association between frailty and complications. Receiver operating characteristic curves were used to explore the predictive ability of frailty.Results: Prevalence of frailty was 26.12%. Significant differences were observed between the frail and non-frail groups with respect to age, Activities of Daily Living, Charlson Comorbidity Index, respiratory function, presence of malignancy, serum albumin, prealbumin, and hemoglobin levels (P

Jiabin Liang,1 Bingbing Han,2 Yunhe Zhang,3 Qingfen Yue1 1Department of Gynaecology, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, Henan 471009, People’s Republic of China; 2Department of Reproductive Medicine, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, Henan 471009, People’s Republic of China; 3Department of Obstetrics and Gynecology, China Meitan General Hospital, Beijing 100028, People’s Republic of China Objective: The present study aimed to investigate the expression of Numb in ovarian cancer tissues and to assess the effect of Numb on cell proliferation, invasion, and EMT in ovarian cancer. Methods: Real-time PCR and Western blotting were used to detect the mRNA and protein expression of Numb, PAK1, β-catenin, and epithelial–mesenchymal transition (EMT)-related proteins. MTT was employed to check the effect of Numb on proliferation of ovarian cancer cells. Transwell assay was performed to examine the functions of Numb and PAK1 on migration and invasion of ovarian cancer cells. Results: The Numb expression was significantly downregulated while PAK1 and β-catenin were significantly upregulated in both ovarian cancer tissues and cell lines. Silencing of Numb promoted cell proliferation, migration, invasion, and EMT in ovarian cancer cell lines while overexpressed Numb reversed the above effects. Moreover, the EMT process induced by the inhibition of Numb was regulated through Numb-mediated PAK1/β-catenin signaling pathway. Conclusion: Numb was downregulated and associated with cell proliferation, invasion, and EMT in ovarian cancer through regulating PAK1/β-catenin signaling, providing a novel potential biomarker and potential therapeutic target for ovarian cancer. Keywords: Numb, ovarian cancer, PAK1, β-catenin, invasion, EMT

Lifeng Zhao,1 Xuesong Li,2 Guanying Liu,1 Baoling Han,1 Jian Wang,1 Xia Jiang1 1Department of Endocrinology, Tianjin First Center Hospital, Tianjin, China; 2First Center Clinical Medical Institute, Tianjin Medical University, Tianjin, China Objective: Recent controversial evidence suggests that maternal diabetes may increase the risk of attention deficit and hyperactivity disorder (ADHD) in offspring. To examine this potential association, a systematic literature search and meta-analysis was performed. Methods: OR or risk ratio (RR) from each study was obtained and combined for evaluating the risk. Six cohort studies and three case-control studies were included in the present study. Results: The meta-analysis of the highly heterogeneous case-control studies did not find significant association between maternal diabetes and ADHD risk (OR: 1.20, 95% CI: 0.96–1.49). The combining of the cohort studies demonstrated that offspring of diabetic mothers were at higher risk of ADHD (RR: 1.40, 95% CI: 1.27–1.54); however, publication bias was identified. When exposure was specified as gestational diabetes mellitus (GDM), GDM exposure increased the risk of ADHD for children by 164% (95% CI: 1.25–5.56) in a Caucasian population. Neither heterogeneity nor publication bias was detected. Conclusion: Maternal diabetes, especially GDM, is probably a risk factor for ADHD in the Caucasian population. More studies based on large sample size and different ethnicities are needed to confirm this association. Keywords: maternal diabetes, attention deficit hyperactivity disorder, meta-analysis

Le Wang,1,2,* Chaoqi Zhang,1,3,* Zhen Zhang,1 Bing Han,4 Zhibo Shen,1 Lifeng Li,1,3 Shasha Liu,1 Xuan Zhao,1 Fanglei Ye,2 Yi Zhang1,3,5 1Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China; 2Department of Otology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China; 3Cancer Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China; 4Department of General ICU, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China; 5Henan Key Laboratory for Tumor Immunology and Biotherapy, Zhengzhou, Henan 450052, China *These authors contributed equally to this work Background: There is a growing recognition that tumor-associated macrophages (TAMs) are recruited to the glioma environment, facilitating tumor proliferation and migration by creating an immunosuppressive microenvironment. CD68 has been widely reported as a specific marker of TAMs in cancer. Purpose: To clarify the role of CD68 in glioma, we investigated its function at the transcriptome level and relationship with clinical practice. Patients and methods: In total, 325 RNA-seq data from Chinese Glioma Genome Atlas (CGGA) and 697 RNA-seq data from The Cancer Genome Atlas (TCGA) network were enrolled in this study. CD68-specific findings were further analyzed with R language, and the prognostic impacts were validated through analyzing the overall survival (OS). Results: CD68 showed a positive correlation with the WHO grade of malignancy in glioma. Meanwhile, CD68 was predominantly expressed in IDH wide type and mesenchymal subtype. Gene ontology (GO) analysis revealed that CD68-related genes were closely related to inflammatory response and immune response. Moreover, seven cultures of metagenes further confirmed that CD68 was a specific marker for macrophages in inflammatory response and played an important role in suppressing T-cell-mediated immunity. The Pearson correlation test suggested that CD68 showed robust correlation with other markers of macrophages and immune checkpoints, including PD-1 and TIM-3. Clinically, a high expression level of CD68 in tumors exhibited a poor survival in glioma patients. Conclusion: Our results demonstrated that CD68 acted as an immune suppressor and contributed to glioma progression in the tumor microenvironment. These findings may expand our understanding of CD68-specific clinical and immune features in glioma. Keywords: CD68, tumor-associated macrophage, glioma, immunotherapy