Southern Medical University 19 articles published in JoVE Behavior Laparoscopic Anterior Right Hepatectomy: A Single-Center Experience Cheng Zhang1, Chang Li1, Chunming Wang1, Lei Cai1, Guolin He1, Shunjun Fu1, Mingxin Pan1 1Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University Here we present a step-by-step protocol for conducting laparoscopic anterior right hepatectomy and compare its clinical effects and postoperative outcomes with those of conventional hepatectomies. Analysis of the data of 82 patients with hepatocellular carcinoma revealed that laparoscopic anterior right hepatectomy had better clinical outcomes and survival rates than the conventional hepatectomy. Biology Isolation and Purification of Bacterial Extracellular Vesicles from Human Feces Using Density Gradient Centrifugation Yicong Xue*1, Xixin Huang*1, Zihao Ou*1, Yuanyuan Wu1, Qianbei Li1, Xinyue Huang1, Minghui Wen1, Yan Yang1, Bo Situ1, Lei Zheng1 1Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University This study describes a method to isolate and purify bacterial extracellular vesicles (BEVs) enriched from human feces via density gradient centrifugation (DGC), identifies the physical characteristics of BEVs from morphology, particle size, and concentration, and discusses the potential applications of the DGC approach in clinical and scientific research. Medicine Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue Jinru Song1, Yue Zhang1, Huihui Pan1, Xueyan Xu1, Cheng-Cheng Deng1, Bin Yang1 1Dermatology Hospital, Southern Medical University This study describes an optimized protocol for establishing primary fibroblasts from keloid tissues that can effectively and steadily provide pure and viable fibroblasts. Immunology and Infection Quantitative Polymerase Chain Reaction (qPCR)-Based Rapid Diagnosis of Helicobacter pylori Infection and Antibiotic Resistance Liang Wang*1,2,3, Jin-Xin Lai*1, Yu-Ting Si*1,4, Xu-Xia Cui1,4, Zeeshan Umar1,5, Xiao-Jun Ru1, Xin-Yu Zhang1, Zheng-Kang Li1, Alfred Chin Yen Tay5,6,7,8, Barry J. Marshall5,6,7,8, Guang-Hua Li1, Bing Gu1 1Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, 2Division of Microbiology and Immunology, School of Biomedical Sciences, The University of Western Australia, 3Center for Precision Health, School of Medical and Health Sciences, Edith Cowan University, 4Medical Technology School of Xuzhou Medical University, 5Marshall Laboratory of Biomedical Engineering, School of Medicine, Shenzhen University, 6The Marshall Centre for Infectious Diseases Research and Training, The University of Western Australia, 7Marshall International Digestive Diseases Hospital, Zhengzhou University, 8Marshall Medical Research Center, Fifth Affiliated Hospital of Zhengzhou University The protocol presents a noninvasive method for the rapid diagnosis of Helicobacter pylori stomach infections through the string test and determines its antibiotic resistance to clarithromycin and levofloxacin using quantitative polymerase chain reaction (qPCR). Medicine Plasma Polishing as a New Polishing Option to Reduce the Surface Roughness of Porous Titanium Alloy for 3D Printing Zhiwei Lin*1,2, Lincong Luo*3, Dongxin Lin*4, Yuping Deng4,5, Yang Yang4, Xuecheng Huang6, Tingrui Wu1, Wenhua Huang1,2,4 1Orthopedic Center, Affiliated Hospital of Guangdong Medical University, 2The School of Basic Medical Sciences, Guangdong Medical University, 3The School of Basic Medical Sciences, Fujian Medical University, 4Guangdong Engineering Research Center for Translation of Medical 3D Printing Application, Guangdong Provincial Key Laboratory of Digital Medicine and Biomechanics, National Key Discipline of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, 5Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, 6Shenzhen Hospital of Guangzhou University of Chinese Medicine (Futian) Plasma polishing is a promising surface processing technology, especially suitable for 3D printing of porous titanium alloy workpieces. It can remove semi-molten powders and ablative oxide layers, thereby effectively reducing surface roughness and improving surface quality. Genetics Pre-Implantation Genetic Testing for Aneuploidy on a Semiconductor Based Next-Generation Sequencing Platform Chengming Xu*1, Riqing Wei*2, Hui Lin1, Leiyu Deng1, Li Wang3, Deyang Li4, Honghui Den5, Wensong Qin1, Ping Wen1, Ying Liu1, Yingsong Wu2, Qiang Ma2, Jinliang Duan1 1Centre for Women, Children, and Reproduction, Guangxi Key Laboratory of Metabolic Diseases Research, 2Department of Biopharmaceutics, School of Laboratory Medicine and Biotechnology, Southern Medical University, 3Department of Obstetrics and Gynecology, Chinese PLA General Hospital, 4Clinical Laboratory, Northern Theater Air Force Hospital, 5Guangzhou Darui Reproduction Technology Co., Ltd. The protocol presents the overall in-lab procedures required in pre-implantation genetic testing for aneuploidy on a semiconductor-based next-generation sequencing platform. Here we present the detailed steps of whole genome amplification, DNA fragment selection, library construction, template preparation, and sequencing working flow with representative results. Bioengineering Fabricating Highly Open Porous Microspheres (HOPMs) via Microfluidic Technology Sheng-Chang Luo1,2, Ying Wang3, Ranjith Kumar Kankala1,2, Yu Shrike Zhang4, Ai-Zheng Chen1,2 1Institute of Biomaterials and Tissue Engineering, Huaqiao University, 2Fujian Provincial Key Laboratory of Biochemical Technology, Huaqiao University, 3Affiliated Dongguan Hospital, Southern Medical University, 4Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School The present protocol describes the fabrication of poly(lactic-co-glycolic acid)-based highly open porous microspheres (HOPMs) via the single-emulsion formulation based facile microfluidic technology. These microspheres have potential applications in tissue engineering and drug screening. Medicine Generation and Characterization of Right Ventricular Myocardial Infarction Induced by Permanent Ligation of the Right Coronary Artery in Mice Ruoxi Liao1, Mingyuan He2, Donghong Hu2, Chengzheng Gong1, Huiying Du1, Hairuo Lin2, Huijun Sun3 1The First Clinical College of Dalian Medical University, 2Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 3College of Pharmacy, Dalian Medical University There are several differences between the right and left ventricles. However, the pathophysiology of right ventricular infarction (RVI) has not been clarified. In the present protocol, a reproducible method for RVI mouse model generation is introduced, which may provide a means to explain the mechanism of RVI. Genetics A Virtual Machine Platform for Non-Computer Professionals for Using Deep Learning to Classify Biological Sequences of Metagenomic Data Zhencheng Fang1,2, Hongwei Zhou1,3 1Microbiome Medicine Center, Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, 2Center for Quantitative Biology, Peking University, 3State Key Laboratory of Organ Failure Research, Southern Medical University This tutorial describes a simple method to construct a deep learning algorithm for performing 2-class sequence classification of metagenomic data. Bioengineering Multiphoton Microscopic Observation of Vessels in Mouse Liver Tissue Wen Rongrong*5, Li Ru*1, He Sixiao1, Wang Ziqing1, Huang Junhao1, Zhao Liying2, Tian Zhihui3,4, Ma Qiang1 1Department of Biopharmaceutics, School of Laboratory Medicine and Biotechnology, Southern Medical University, 2Department of General Surgery, Nanfang Hospital, Southern Medical University, 3Department of Stomatology, Nanfang Hospital, Southern Medical University, 4College of Stomatology, Southern Medical University, 5School of Laboratory Medicine and Biotechnology, Southern Medical University In this experiment, a mouse is injected in its tail vein with Rhodamine B isothiocyanate–dextran that can stain blood vessels. After the liver is exposed and fixed, a specific part of the liver can be selected to observe the deep tissue in the living body using multiphoton microscopy. Medicine Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy Yi Lao1, Cankun Zheng1, Hailin Zhu1,2, Hairuo Lin1, Xiaoxia Huang1, Yulin Liao1 1Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 2Department of Geriatrics, Pingxiang People's Hospital of Southern Medical University This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning. Developmental Biology An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo Yunyun Jiang1, Jiahao Chen1, Jin Xu2 1Department of Developmental Biology, School of Basic Medical Sciences, Southern Medical University, 2Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology To test the effect of a chemokine on macrophage recruitment in vivo, the whole mount in situ hybridization was used to detect the ectopic expression of the chemokine, and immunostaining was used to label macrophages. Live imaging was used for real-time observation of macrophage migration. Behavior Application of 3D Printing in the Construction of Burr Hole Ring for Deep Brain Stimulation Implants Jiazhi Chen*1,2, Xinyu Chen*3, Siyuan Lv2, Yuzhen Zhang1, Hao Long1, Kaijun Yang1, Songtao Qi1, Wangming Zhang2, Jun Wang1 1Department of Neurosurgery, Nanfang Hospital, Southern Medical University, 2Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, 3Faculty of Brain Sciences, University of College London Here, we present a protocol to demonstrate 3D printing in the construction of deep brain stimulation implants. Biology Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice Qiancheng Wang1, Kaitong Chen1, Hairuo Lin1, Mingyuan He2, Xiaoxia Huang1, Hailin Zhu1, Yulin Liao1 1Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 2The Third Clinical Medical College, Guangzhou Medical University Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided. Medicine Mechanical Micronization of Lipoaspirates for Regenerative Therapy Huidong Zhu1, Jinbo Ge1, Xihang Chen1, Feng Lu*1, Junrong Cai*1 1Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University Here, we present a protocol to obtain stromal vascular fraction from adipose tissue through a series of mechanical processes, which include emulsification and multiple centrifugations. Developmental Biology A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome Ping Xue1, Zhiqiang Wang1, Xiaomin Fu1,2, Junjiang Wang1,3, Gopika Punchhi1, Andrew Wolfe1,4, Sheng Wu1,4,5 1Department of Pediatrics, Johns Hopkins University School of Medicine, 2Department of Health, Beijing Military General Hospital, 3Southern Medical University, 4Department of Molecular and Cellular Physiology, Johns Hopkins University School of Medicine, 5Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine We describe the development of a lean PCOS-like mouse model with dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams. Neuroscience Auditory Brainstem Response and Outer Hair Cell Whole-cell Patch Clamp Recording in Postnatal Rats Aoshuang Chang1,2, Cuixian Li1, Jianfeng Huang1,3, Wenlu Pan1, Yinghong Tian4, Jie Tang1 1Department of Physiology, School of Basic Medical Sciences, Southern Medical University, 2School of Basic Medical Sciences, Guizhou Medical University, 3Department of Laboratory Medicines, Guangzhou General Hospital of Guangzhou Military Region, Southern Medical University, 4Experiment Teaching Center, School of Basic Medical Sciences, Southern Medical University This protocol describes methods to record the auditory brainstem response from postnatal rat pups. To examine the functional development of outer hair cells, the experimental procedure of whole-cell patch clamp recording in isolated outer hair cells is described step-by-step. Developmental Biology Use of a Recombinant Mosquito Densovirus As a Gene Delivery Vector for the Functional Analysis of Genes in Mosquito Larvae Pei-Wen Liu1, Jia-Bao Xu1, Yun-Qiao Dong2, Xiao-Guang Chen1, Jin-Bao Gu1 1Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, 2 We report using an artificial intronic small RNA expression strategy to develop a non-defective recombinant Aedes aegypti densovirus (AaeDV) in vivo delivery system. A detailed procedure for the construction, packaging, and quantitative analysis of the rAaeDV vectors as well as for larval infection is described. Immunology and Infection Intravital Imaging of Neutrophil Priming Using IL-1β Promoter-driven DsRed Reporter Mice Yi Yao1,2, Yun Liu1,3, Akira Takashima1 1Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, 2Department of Internal Medicine, Yale University School of Medicine, 3Department of Pathophysiology, Southern Medical University (China) This current protocol employs fluorescent reporters, in vivo labeling, and intravital imaging techniques to enable monitoring of the dynamic process of neutrophil priming in living animals.