Central South University 18 articles published in JoVE Neuroscience Rat Model of Closed-Head Mild Traumatic Injury and its Validation Yishu Liu*1, Tingting Wang*1, Changquan Zhang1, Jifeng Cai1 1Department of Forensic Science, School of Basic Medical Science, Central South University Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages. Immunology and Infection Evaluation of a Reliable Biomarker in a Cecal Ligation and Puncture-Induced Mouse Model of Sepsis Li Yaozhen1, Wu Kemin1, Ji Xiaoyu1, Ouyang Yang1, Tang Hongying1, Pan Baihong1 1Department of Vascular Surgery, Xiangya Hospital, Central South University This protocol presents the operative details of cecal ligation and puncture (CLP) in a mouse model of sepsis. CLP is one of the most widely used techniques to create an animal model of sepsis. Therefore, a standardized CLP protocol is required for the attainment of reliable research results. Medicine Ultrasound-Guided Induced Pluripotent Stem Cell-Derived Cardiomyocyte Implantation in Myocardial Infarcted Mice Xun Wu1, Kele Qin1, Di Wang2, Kun Xiang1, Jun Peng2, Jianjun Guo3, Jinfu Yang1,2, Chengming Fan1,2,3 1Department of Cardiovascular Surgery, Second Xiangya Hospital, Central South University, 2Hunan Provincial Key Laboratory of Cardiovascular Research, 3Hunan Fangsheng Pharmaceutical Co., Ltd. Ultrasound-guided cell delivery around the site of myocardial infarction in mice is a safe, effective, and convenient way of cell transplantation. Medicine In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter Zhaokai Li*1,2, Liu Liu*1, Zhong Wang1 1Department of Cardiac Surgery, Cardiovascular Center, The University of Michigan, Ann Arbor, 2Department of Cardiovascular Medicine, Xiangya Hospital, Central South University We describe here, the establishment and application of an Tg(Myh6-cre)1Jmk/J /Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze/J (referred to as αMHC-Cre/Rosa26A-Flox-Stop-Flox-GCaMP3 below) mouse reporter line for cardiac reprogramming assessment. Neonatal cardiac fibroblasts (NCFs) isolated from the mouse strain are converted into induced cardiomyocytes (iCMs), allowing for convenient and efficient evaluation of reprogramming efficiency and functional maturation of iCMs via calcium (Ca2+) flux. Medicine Cox-Maze IV Procedure Concomitant with Valvular Surgery In Situs Inversus Dextrocardia: A Single-Center Experience in China Cheng Luo*1, Chengming Fan*1, Hao Zhang1, Long Song1, Yuhong Liu1, Liming Liu1 1Department of Cardiovascular Surgery, the Second Xiangya Hospital, Central South University We summarize the Cox-Maze IV procedure concomitant with valvular surgery performed in patients with situs inversus dextrocardia at this institution. Biology A Bacterial Oral Feeding Assay with Antibiotic-Treated Mosquitoes Xinyi Liu1,2, Si Wu1,2, Wenqian Li1,2, Meihong Zhang1,2, Yan Wu1,2, Ning Zhou3, Pa Wu1,2 1Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, College of Life Science, Hunan Normal University, 2State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Provincial Key Laboratory for Microbial Molecular Biology, College of Life Science, Hunan Normal University, 3Department of Infectious Diseases, the Second Xiangya Hospital, Central South University This article presents a protocol to investigate the effect of individual mosquito gut bacteria, including isolation and identification of mosquito midgut cultivable microbes, antibiotic depletion of mosquito gut bacteria, and reintroduce one specific bacteria species. Immunology and Infection Isolation of Mouse Kidney-Resident CD8+ T cells for Flow Cytometry Analysis Wei Liao1,2, Chaoyu Ma1, Nu Zhang1 1Department of Microbiology, Immunology and Molecular Genetics, School of Medicine, University of Texas Health Science Center at San Antonio, 2Department of Dermatology, Hunan Key Laboratory of Medical Epigenomics, The Second Xiangya Hospital, Central South University Following viral infection, kidney harbors a relatively large number of CD8+ T cells and offers an opportunity to study non-mucosal TRM cells. Here, we describe a protocol to isolate mouse kidney lymphocytes for flow cytometry analysis. Immunology and Infection Optimization of the Cuff Technique for Murine Heart Transplantation Yunhan Ma*1,2, Baiyi Xie*1,2, Helong Dai*3,4,5, Chenxi Wang6, Shujiao Liu6, Tianshu Lan7, Shuangyue Xu1,2, Guoliang Yan1,2,6, Zhongquan Qi1,2,8 1Organ Transplantation Institute, School of Medicine, Xiamen University, 2Fujian Key Laboratory of Organ and Tissue Regeneration, 3Department of Kidney Transplantation, Center of Organ Transplantation, The Second Xiangya Hospital of Central South University, 4Clinical Research Center for Organ Transplantation in Hunan Province, 5Clinical Immunology Center, Central South University, 6School of Medicine, Xiamen University, 7Xiamen Medical College, 8Medical College, Guangxi University We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time. Immunology and Infection Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation Hamad H. Alanazi1,2, Li She1,3, Xiao-Dong Li1 1Department of Microbiology, Immunology and Molecular Genetics, Long School of Medicine, University of Texas Health San Antonio, 2Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, 3Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University Environmental allergens such as house dust mites (HDM) often contain microbial substances that activate innate immune responses to regulate allergic inflammation. The protocol presented here demonstrates the identification of dsRNA species in HDM allergens and characterization of their immunogenic activities in modulating eosinophilic lung inflammation. Behavior Assessment of Sexual Behavior of Male Mice Zi-Wei Liu1, Ning Jiang2, Xue Tao2, Xiao-Ping Wang1, Xin-Min Liu2, Shui-Yuan Xiao3 1Department of Psychiatry, The Second Xiangya Hospital, Central South University, National Clinical Research Center for Mental Disorders and National Technology Institute on Mental Disorders, Hunan Key Laboratory of Psychiatry and Mental Health, 2Research Center for Pharmacology & Toxicology, Institute of Medicinal Plant Development (IMPLAD), Chinese Academy of Medical Sciences and Peking Union Medical College, 3Department of Social Medicine and Health Management, Xiangya School of Public Health, Central South University This article describes how to perform sexual behavior tests in male mice. Cancer Research Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis Xianquan Zhan1,2,3,4, Huanni Li5, Shehua Qian1,2,3, Xiaohan Zhan1,2,3, Na Li1,2,3 1Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, 2Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, 3State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, 4National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, 5Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University This article presents a protocol of differential-speed centrifugation in combination with density gradient centrifugation to separate mitochondria from human ovarian cancer tissues and control ovarian tissues for quantitative proteomics analysis, resulting in a high-quality mitochondrial sample and high-throughput and high-reproducibility quantitative proteomics analysis of a human ovarian cancer mitochondrial proteome. Medicine Blood Collection Through Subclavian Vein Puncture in Mice Hui Yang*1, Chenlu Wu*1, Fengjiao Liu2, Mi Wang1, Pu Zou1, Yuhu He1, Qiming Liu1, Qin Zhou3, Shenghua Zhou1 1Department of Cardiology, The Second Xiangya Hospital, Central South University, 2Department of Cardiology, The Xiangya Hospital, Central South University, 3Department of General Surgery, The Second Xiangya Hospital, Central South University Here, we present a protocol to take blood samples from the subclavian vein of mice. Cancer Research Elastic Staining on Paraffin-embedded Slides of pT3N0M0 Gastric Cancer Tissue Guang Lei*1, Haiyan Yang*1, Ting Hong2, Xiangyu Zhang1, Nong Yang1, Yongchang Zhang1 1Department of Medical Oncology, Lung Cancer and Gastrointestinal Unit, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, 2Hunan ClinicaI Research Center in Gynecologic Cancer, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University Here, we present a protocol of elastic staining to identify elastic fibers in pT3N0M0 gastric cancer tissues on formalin-fixed, paraffin-embedded sections. Subsequently, we describe a method to determine whether tumor cells invade beyond the elastic lamina. Medicine Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats Hui Yang1, Pu Zou1, Jingyuan Chen1, Gaoxing Shi1, Chenlu Wu1, Mi Wang1, Qin Zhou2, Shenghua Zhou1 1Department of Cardiology, The Second Xiangya Hospital, Central South University, 2Department of General Surgery, The Second Xiangya Hospital, Central South University Here, we present a protocol to collect blood samples from the rat subclavian vein. Cancer Research Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome Xianquan Zhan1,2,3,4, Yuda Huang1,2,3, Ying Long1,2,3 1Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, 2Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, 3State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, 4The State Key Laboratory of Medical Genetics, Central South University Here, we present a two-dimensional gel electrophoresis (2DE) coupled with mass spectrometry (MS) to separate and identify human pituitary adenoma tissue proteome, which presents a good and reproducible 2DE pattern. Many proteins are observed in each 2DE spot when analyzing complex cancer proteome with the use of high-sensitivity MS. Neuroscience Partial Optic Nerve Transection in Rats: A Model Established with a New Operative Approach to Assess Secondary Degeneration of Retinal Ganglion Cells Fancheng Yan1, Sailiang Guo2, Yijie Chai2, Lan Zhang3, Kegao Liu3, Qingjun Lu4, Ningli Wang4, Shuning Li3 1Aier School of Ophthalmology, Central South University, Changsha, China, 2Institute of Immunology, Tsinghua University School of Medicine, Beijing, China, 3Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University; Beijing Ophthalmology & Visual Sciences Key Laboratory, Beijing, China, 4Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University; Beijing Ophthalmology & Visual Sciences Key Laboratory, Beijing, China Secondary degeneration of retinal ganglion cells (RGCs) occurs commonly in glaucoma. This study describes an innovative operative approach for partial optic nerve transection. The use of this space-saving operative approach extends the model's application range, and allows exploration of secondary injury mechanisms in RGCs in a new way. Biology Isolation and Primary Culture of Mouse Aortic Endothelial Cells Jie-Mei Wang1, Alex F. Chen2, Kezhong Zhang1 1Center for Molecular Medicine and Genetics, Wayne State University, 2Third Xiangya Hospital and the Institute of Vascular Disease and Translational Medicine, Central South University The vascular endothelial cells play a significant role in many important cardiovascular disorders. This article describes a simple method to isolate and expand endothelial cells from the mouse aorta without using any special equipment. Our protocol provides an effective means of identifying mechanisms in endothelial cell physiopathology. Medicine Technical Aspects of the Mouse Aortocaval Fistula Kota Yamamoto1,2, Xin Li1,3, Chang Shu3, Tetsuro Miyata2, Alan Dardik1,4 1The Department of Surgery and the Interdepartmental Program in Vascular Biology and Therapeutics, Yale University, 2Department of Vascular Surgery, The University of Tokyo, 3Department of Vascular Surgery, Central South University, 4VA Connecticut Healthcare Systems The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.