Huazhong University of Science and Technology 24 articles published in JoVE Biology Dual-Dye Optical Mapping of Hearts from RyR2R2474S Knock-In Mice of Catecholaminergic Polymorphic Ventricular Tachycardia Yangpeng Li*1,2, Jun Yang*1, Rui Zhang1, Tangting Chen1, Shiyu Zhang1, Yuqing Zheng1, Qiang Wen3, Tao Li1, Xiaoqiu Tan1,2, Ming Lei1,4, Xianhong Ou1 1Key Laboratory of Medical Electrophysiology of Ministry of Education, Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease, Institute of Cardiovascular Research, Southwest Medical University, 2Department of Cardiology, the Affiliated Hospital of Southwest Medical University, 3Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 4Department of Pharmacology, University of Oxford This protocol introduces dual-dye optical mapping of mouse hearts obtained from wild-type and knock-in animals affected by catecholaminergic polymorphic ventricular tachycardia, including electrophysiological measurements of transmembrane voltage and intracellular Ca2+ transients with high temporal and spatial resolution. Medicine Isolation and Identification of Limbal Niche Cells Guanyu Su1, Wei Wang1, Lingjuan Xu1, Rong Liu1, Yongyao Tan1, Bihui Jin1, Chunxiu You1, Xiao Zhou1, Yihong Xiong2, Huatao Xie3, Guigang Li1 1Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2Institute of Medicine Nursing, Hubei University of Medicine, 3Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology Here, we present a protocol to isolate and identify the human limbal niche cells. Engineering A Flexible Wearable Supernumerary Robotic Limb for Chronic Stroke Patients Hongge Ru1, WeiJian Gao2, Weixuan Ou2, Xingyue Yang1, Andong Li1, Zhongzheng Fu1, Jun Huo1, Bo Yang1, Yanzhao Zhang3, Xiling Xiao3, Zhaohui Yang3, Jian Huang1 1Key Laboratory for Image Processing and Intelligent Control of Education Ministry of China, School of Artificial Intelligence and Automation, Huazhong University of Science and Technology, 2School of Mechanical Science and Engineering, Huazhong University of Science and Technology, 3Wuhan Union Hospital, Huazhong University of Science and Technology Tongji Medical College First Clinical College Union Hospital This protocol introduces a flexible wearable supernumerary robotic limb tailored to assist in finger rehabilitation for stroke patients. The design incorporates a bending sensor to facilitate seamless human-robot interaction. Validation through experiments involving both healthy volunteers and stroke patients underscores the efficacy and dependability of the proposed study. Medicine Clinical Application of 24 G Cannula Needle and 3-0 Polypropylene Suture in Vas Deferens Exploration Jin Wang*1, Wenjia Li*2, Yao He1, Yuanbin Xia1, Lin Sun1, Youpeng Zhang1, Zhaohui Zhu1 1Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology This protocol presents the clinical application of a 24 G cannula and 3-0 polypropylene suture as a simple and effective method for the exploration of the vas deferens. Cancer Research Development of a 68Gallium-Labeled D-Peptide PET Tracer for Imaging Programmed Death-Ligand 1 Expression Lulu Zhang*1, Siqi Zhang*2, Wenyu Wu1, Xingkai Wang2, Jieting Shen2, Dongyuan Wang4, Kuan Hu*2,3, Ming-Rong Zhang*3, Feng Wang*1, Rui Wang*2 1Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, 2State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 3Department of Advanced Nuclear Medicine Sciences, Institute for Quantum Medical Science, National Institutes for Quantum Science and Technology, 4Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology This study developed a noninvasive and real-time method to evaluate the distribution of programmed death-ligand 1 in the whole body, based on positron emission tomographic imaging of [68Ga] D-dodecapeptide antagonist. This technique has advantages over conventional immunohistochemistry and improves the efficiency of identifying appropriate patients who will benefit from immune checkpoint blockade therapy. Engineering A Teleoperated Robotic System-Assisted Percutaneous Transiliac-Transsacral Screw Fixation Technique Binbin Liu1, Fei Xu1, Yang Liu1, Tianqi Wang2, Yajing Cao2, Zehang Zheng1, Hanqing Xu1, Chuang Huang1, Zhengqiang Luo1 1Department of Orthopedics, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, 2Wuhan United Imaging Healthcare Surgical Technology Co. Ltd. Teleoperated robotic system-assisted percutaneous transiliac-transsacral screw fixation is a feasible technique. Screw channels can be implemented with high accuracy owing to the excellent freedom of movement and stability of the robotic arms. Biology A Modified Technique for Transverse Aortic Constriction in Mice Kudusi Abuduwufuer*1,2, James Jiqi Wang*1,2, Huihui Li*1,2, Chen Chen1,2 1Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2Hubei Key Laboratory of Genetics and Molecular Mechanism of Cardiological Disorders, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology The present protocol describes a modified and simplified technique with a minimally invasive transverse aortic constriction (TAC) procedure using a self-made retractor. This procedure can be conducted without a ventilator or microscope and introduces pressure overload, eventually leading to cardiac hypertrophy or heart failure. Genetics Assisted Selection of Biomarkers by Linear Discriminant Analysis Effect Size (LEfSe) in Microbiome Data Fang Chang*1, Shishi He*2, Chenyuan Dang3 1Marine Resources Research Centre, Tianjin Research Institute for Water Transport Engineering, M.O.T., 2Key Laboratory of Regional Energy Systems Optimization, Ministry of Education, College of Environmental Science and Engineering, North China Electric Power University, 3School of Environmental Science and Engineering, Huazhong University of Science and Technology LEfSe (LDA Effect Size) is a tool for high-dimensional biomarker mining to identify genomic features (such as genes, pathways, and taxonomies) that significantly characterize two or more groups in microbiome data. Neuroscience A Stably Established Two-Point Injection of Lysophosphatidylcholine-Induced Focal Demyelination Model in Mice Xiao-Wei Pang1, Man Chen1, Yun-Hui Chu1, Yue Tang1, Chuan Qin1, Dai-Shi Tian1 1Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology The present protocol describes a two-point injection of lysophosphatidylcholine via a stereotaxic frame to generate a stable and reproducible demyelination model in mice. Medicine Interphase Fluorescence in situ Hybridization of Bone Marrow Smears of Multiple Myeloma Yalan Yu*1, Hui Shen*1, Li Liu1, Ping Luo1, Sanyun Wu1, Jing He1,2, Xiqin Tong1, Yufeng Shang1, Liang Shao1, Fuling Zhou1 1Department of Hematology, Zhongnan Hospital of Wuhan University, 2Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology Here, we present a protocol for improving the success of interphase fluorescence in situ hybridization detection on bone marrow smears from multiple myeloma patients. Neuroscience Isolation of Mouse Primary Microglia by Magnetic-Activated Cell Sorting in Animal Models of Demyelination Hang Zhang1, Sheng Yang1, Man Chen1, Dai-Shi Tian1, Chuan Qin1 1Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Here, we present a protocol to isolate and purify primary microglia in animal models of demyelinating diseases, utilizing columnar magnetic-activated cell sorting. Biology Assessing Energy Substrate Oxidation In Vitro with 14CO2 Trapping Chao Song1,2, Arianna Valeri1, Fanxin Long1 1 This protocol describes an easy-to-use method to examine substrate oxidation by tracking 14CO2 production in vitro. Medicine Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice Bingjie Lv*1, Jin Zhou*1, Shaolin He*1, Yuqi Zheng1, Wenlin Yang1, Shangwei Liu1, Chengxing Liu1, Boyuan Wang1, Dazhu Li1, Jibin Lin1 1Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology Here we describe a simple and reproducible method that can induce myocardial infarction or myocardial ischemia-reperfusion injury in mice by precision ligation of the left anterior descending coronary artery through micromanipulation. Neuroscience Free-floating Immunostaining of Mouse Brains Longlong Tu1, Nan Zhang1,2,3, Kristine M Conde1, Jonathan C Bean1, Chunmei Wang1, Yong Xu1,4 1USDA/ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 2Department of Endocrinology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 3Hubei Provincial Clinical Research Center for Diabetes and Metabolic Disorder, 4Department of Molecular and Cellular Biology, Baylor College of Medicine This protocol describes an efficient and reproducible approach for mouse brain histological studies, including perfusion, brain sectioning, free-floating immunostaining, tissue mounting, and imaging. Behavior A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice Yi Xie1, Sai-Yue Deng1, Si-Miao Chen1, Xue-Jiao Chen1, Wen-Wen Lai1, Li-Fang Huang1, Li Ba1, Wei Wang1,2, Feng-Fei Ding1,3 1Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China, 2Key Laboratory of Neurological Diseases of Chinese Ministry of Education, The School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000,China, 3Department of Pharmacology, School of Basic Medical Science; State Key Laboratory of Medical Neurobiology, Institutes of Brain Science and Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China Presented here is a protocol for chronic sleep fragmentation (CSF) model achieved by an electrically controlled orbital rotor, which could induce confirmed cognitive deficit and anxiety-like behavior in young wild-type mice. This model can be applied to explore the pathogenesis of chronic sleep disturbance and related disorders. Engineering In Situ Measurement of Vacuum Window Birefringence using 25Mg+ Fluorescence W. H. Yuan1, H. L. Liu1, W. Z. Wei1, Z. Y. Ma1, P. Hao1, Z. Deng1, K. Deng1, J. Zhang1, Z. H. Lu1 1MOE Key Laboratory of Fundamental Physical Quantities Measurement, Hubei Key Laboratory of Gravitation and Quantum Physics, PGMF and School of Physics, Huazhong University of Science and Technology Presented here is a method to measure the birefringence of vacuum windows by maximizing the fluorescence counts emitted by Doppler cooled 25Mg+ ions in an ion trap. The birefringence of vacuum windows will change the polarization states of the laser, which can be compensated by changing the azimuthal angles of external wave plates. Medicine A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice Jie Zhang1,2, Fang Fang1,3, Liang Li1, Haoliang Huang1, Hannah C. Webber1, Yang Sun1,4, Vinit B. Mahajan1,4, Yang Hu1 1Department of Ophthalmology, Stanford University School of Medicine, 2Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 3Department of Ophthalmology, Second Xiangya Hospital of Central South University, 4Department of Ophthalmology, Veterans Affairs Palo Alto Health Care Here, we present a protocol to induce ocular hypertension and glaucomatous neurodegeneration in mouse eyes by intracameral injection of silicone oil and the procedure for silicone oil removal from the anterior chamber to return elevated intraocular pressure to normal. Neuroscience Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5 Shuiqing Chi1, Mijing Fang1, Kang Li1, Li Yang2, Shao-tao Tang1 1Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2 This protocol describes the process of immunostaining rectal suction biopsies for calretinin, S100 protein, and protein gene product 9.5. This novel adjuvant diagnostic method for Hirschsprung's disease has preferable sensitivity and specificity rates. Behavior Modified Blood Collection from Tail Veins of Non-anesthetized Mice with a Vacuum Blood Collection System and Eyeglass Magnifier Xiaoyan Liu*1, Huanhuan Li*1, Jie Wu1, Yijun Yu1, Mingjing Zhang1, Wusong Zou1, Ye Gu1 1Heart Center, Wuhan Fourth Hospital, Puai Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology This study reports blood sampling from tail vein in mice using a vacuum extraction tube system with eyeglass magnifier. Our method is easy to practice and could be used for repeat blood sampling in mice. Engineering Nerve Stem Cell Differentiation by a One-step Cold Atmospheric Plasma Treatment In Vitro Zilan Xiong*1, Shasha Zhao*2, Xu Yan3 1State Key Laboratory of Advanced Electromagnetic Engineering and Technology, Huazhong University of Science and Technology, 2College of Life Science and Health, Wuhan University of Science and Technology, 3Department of Pathophysiology, Beijing Neurosurgical Institute/Beijing Tiantan Hospital, Capital Medical University This protocol aims to provide detailed experimental steps of a cold atmospheric plasma treatment on neural stem cells and immunofluorescence detection for differentiation enhancement. Bioengineering Light-sheet Fluorescence Microscopy for the Study of the Murine Heart Yichen Ding1, Zachary Bailey2, Victoria Messerschmidt2, Jun Nie3, Richard Bryant2, Sandra Rugonyi4, Peng Fei3, Juhyun Lee1,2, Tzung K. Hsiai1 1Department of Bioengineering, University of California Los Angeles, 2Department of Bioengineering, University of Texas at Arlington, 3School of Optical and Electronic Information, Huazhong University of Science and Technology, 4Department of Biomedical Engineering, OSHU This study uses a dual-sided illumination light-sheet fluorescence microscopy (LSFM) technique combined with optical clearing to study the murine heart. Biochemistry Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers Huijuan You*1, Shimin Le*2,3, Hu Chen2,4, Linyan Qin1, Jie Yan2,4,5 1School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, 2Mechanobiology Institute, National University of Singapore, 3Department of Physics, National University of Singapore, 4Research Institute for Biomimetics and Soft Matter, Fujian Provincial Key Lab for Soft Functional Materials Research, Department of Physics, Xiamen University, 5Center for Bioimaging Sciences, National University of Singapore A single-molecule magnetic tweezers platform to manipulate G-quadruplexes is reported, which allows for the study of G4 stability and regulation by various proteins. Medicine Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect Bo Zhang1,2, Jonathan P. Davis1, Mark T. Ziolo1 1Department of Physiology & Cell Biology, Davis Heart and Lung Research Institute, The Ohio State University, 2Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates. Bioengineering Porous Silicon Microparticles for Delivery of siRNA Therapeutics Jianliang Shen*1,2, Xiaoyan Wu*1,3, Yeonju Lee1, Joy Wolfram1,4, Zhizhou Yang1, Zong-Wan Mao2, Mauro Ferrari1,5, Haifa Shen1,6 1Department of Nanomedicine, Houston Methodist Research Institute, 2MOE Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry and Chemical Engineering, Sun Yat-sen University, 3Pediatrics Department of Union Hospital, Huazhong University of Science and Technology, 4CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience & Technology of China, 5Department of Medicine, Weill Cornell Medical College, 6Department of Cell and Developmental Biology, Weill Cornell Medical College Delivery remains the main challenge for the therapeutic implementation of small interfering RNA (siRNA). This protocol involves the use of a multifunctional and biocompatible siRNA delivery platform, consisting of arginine and polyethylenimine grafted porous silicon microparticles.
