Wenzhou Medical University 10 articles published in JoVE Medicine Cognitive Function and Upper Limb Rehabilitation Training Post-Stroke Using a Digital Occupational Training System Zhao Yao1, Tianfang Zhang2, Fenfen Chen1,3, Weiyi Shi2, Jinjin zheng2, Ziwei Zhang2, Zuobing Chen1,2 1The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, 2Department of Rehabilitation Medicine, The First Affiliated Hospital, School of Medicine, Zhejiang University, 3Department of Rehabilitation Medicine, Taizhou Hospital Affiliated to Wenzhou Medical University The current protocol outlines how the VR-based digital occupational training system enhances the rehabilitation of patients with cognitive impairment and upper limb dysfunction following a stroke. Neuroscience Targeted Knockdown of Genes in the Choroid Plexus Yuwen Yang*1, Chenxing Qi*1, Lanxin Hu1, Cheng Zheng1, Xuhang Li1, Wu Zheng1, Yiyun Weng2, Haiyan Lin3,4 1State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University, 2Department of Neurology, the First Affiliated Hospital of Wenzhou Medical University, 3Rehabilitation Medicine Center, The Second Affiliated Hospital of Wenzhou Medical University, 4Integrative & Optimized Medicine Research Center, China-USA Institute for Acupuncture and Rehabilitation, Wenzhou Medical University Here, we describe a method to selectively alter gene expressions in the choroid plexus while avoiding any impact in other brain areas. Biology Visualizing Mitophagy with Fluorescent Dyes for Mitochondria and Lysosome Bilin Liu1,2, Anqi Li1, Yuan Qin3, Lei Chen1, Meng Gao1, Guohua Gong1,2 1Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, 2Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 3Department of Pharmacy, Shanghai East Hospital, Tongji University Mitophagy is the primary mechanism of mitochondrial quality control. However, the evaluation of mitophagy in vivo is hindered by the lack of reliable quantitative assays. Presented here is a protocol for the observation of mitophagy in living cells using a cell-permeant green-fluorescent mitochondria dye and a red-fluorescent lysosome dye. Neuroscience In Vivo Methods to Assess Retinal Ganglion Cell and Optic Nerve Function and Structure in Large Animals Qian Ye*1, Zhonghao Yu*1, Tian Xia*1, Shengjian Lu1, Jiaying Sun1, Mengyun Li1, Yu Xia1, Si Zhang1, Wencan Wu1, Yikui Zhang1 1The Eye Hospital, School of Ophthalmology & Optometry, Wenzhou Medical University Here we demostrate several in vivo tests (flash visual evoked potential, pattern electroretinogram and optic coherence tomography) in goat and rhesus macaque to understand the structure and function of the optic nerve and its neurons. Immunology and Infection Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin Di Zhang*1,2, Kefang Liu*3,4, Dan Lu*2,5, Pengyan Wang1,2, Qingxu Zhang1,2, Peipei Liu2, Yingze Zhao2, Yan Chai4, Jianxin Lyu1, Jianxun Qi4, William J. Liu1,2 1School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 2NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 3Faculty of Health Sciences, University of Macau, 4CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 5Savaid Medical School, University of Chinese Academy of Sciences The protocol describes experimental methods to obtain stable major histocompatibility complex (MHC) class I through potential β2-microglobulin (β2m) substitutions from different species. The structural comparison of MHC I stabilized by homologous and heterologous β2m were investigated. Medicine Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets Yuanzheng Peng*1, Zhicheng Zou*1, Jiao Chen1, Hancheng Zhang1, Ying Lu1, Rito Bittino2, Hongxing Fu3, David K. C. Cooper4, Shan Lin1, Mengtao Cao1, Yifan Dai5, Zhiming Cai1, Lisha Mou1 1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Health Science Center, 2Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, 3School of Pharmaceutical Sciences, Wenzhou Medical University, 4Xenotransplantation Program/Department of Surgery, University of Alabama at Birmingham, 5Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed. Biology A Method for Islet Transplantation to the Omentum in Mouse Ying Lu*1, Shangyou Zou*1, Suzanne Bertera*2, Rita Bottino2, David K.C. Cooper3, Zhengzhao Liu1, Yi Huang1, Chengjun Wang1, Chungu Hong1, Tian He1, Hancheng Zhang1, Qin Huo4, Hongxing Fu5, Zhiming Cai1, Lisha Mou1 1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen University School of Medicine, Shenzhen University Health Science Center, 2Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, 3Xenotransplantation Program/Department of Surgery, The University of Alabama at Birmingham, 4College of Life Science and Oceanography, Shenzhen University, 5School of Pharmcy, Wenzhou Medical College A method for the omental transplantation of islets in a mouse is introduced. The isolated islets are mixed with hydrogel and the mixture is placed into the omental pouch of a diabetic mouse. Then, the blood glucose is monitored, and immuno-histochemical analysis is performed. Immunology and Infection Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice Yongli Zhang*1,2, Hangjie Zhang*2, Wenqiang Ma*3, Kefang Liu1,2, Min Zhao4, Yingze Zhao2, Xuancheng Lu5, Fuping Zhang6, Xiangdong Li3, George F. Gao1,2,4,6, William J. Liu1,2 1School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 2NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 3State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, 4Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science, Chinese Academy of Sciences, 5Laboratory Animal Center, Chinese Center for Disease Control and Prevention, 6CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation. Developmental Biology In Vitro Growth of Mouse Preantral Follicles Under Simulated Microgravity Shen Zhang1, Yonggen Wu1, Yimin Weng2, Zhihui Xu1, Wenmin Chen3, Dahan Zheng4, Wei Lin5, Jun Liu6, Ying Zhou1,7 1Reproductive Medicine Center, The First Affiliated Hospital of Wenzhou Medical University, 2Department of Orthopaedics, The Second Affiliated Hospital of Wenzhou Medical University, 3Department of Obstetrics, The First Affiliated Hospital of Wenzhou Medical University, 4School of Laboratory Medicine and Life Science, Wenzhou Medical University, 5School of Pharmaceutical Science, Wenzhou Medical University, 6Stem Cells and Genetic Engineering Group, AgriBioscience Research Centre, Department of Economic Development, Jobs, Transport and Resources, 7Department of Histology and Embryology, Wenzhou Medical University A highly promising technique to generate tissue constructs without using matrix is to culture cells in a simulated microgravity condition. Using a rotary culture system, we examined ovarian follicle growth and oocyte maturation in terms of follicle survival, morphology, growth, and oocyte function under the simulated microgravity condition. Developmental Biology Rapid Acquisition of 3D Images Using High-resolution Episcopic Microscopy Haochuan Zhang*1,2,3, JunGang Huang*2,3,4, Xin Liu2,3, Ping Zhu4, Zhongrong Li1, Xue Li2,3 1Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University, 2 We describe a detailed protocol using high-resolution episcopic microscopy to acquire three-dimensional (3D) images of mouse embryos. This improved protocol utilizes a modified tissue preparation method to enhance penetration of the fluorescent dye, thereby permitting morphometric analysis of both small and large-sized specimens.
