Zhejiang University School of Medicine 14 articles published in JoVE Bioengineering Cell Patterning Using Magnetic-Archimedes Strategy Xuhao Zhou1,2,3, Miribani Maitusong1,2,3, Tanchen Ren1,2,3, Yan Wu1,2,3 1Department of Cardiology of The Second Affiliated Hospital, Zhejiang University School of Medicine, 2State Key Laboratory of Transvascular Implantation Devices, 3Cardiovascular Key Laboratory of Zhejiang Province This protocol describes an ink-free, label-free, substrate-independent, high-throughput cell patterning method based on the Magnetic Archimedes effect. Biology Comparison of Two Representative Methods for Differentiation of Human Induced Pluripotent Stem Cells into Mesenchymal Stromal Cells Fangqin Wang1, Langping Gao2,3, Xudong Fu4, Qintao Yan2, Lidan Hu1,2, Jianhua Mao2,3 1Department of Nephrology, The Children's Hospital, Zhejiang University School of Medicine, 3Zhejiang University School of Medicine, 4Liangzhu Laboratory, Zhejiang University This protocol describes and compares two representative methods for differentiating hiPSCs into mesenchymal stromal cells (MSCs). The monolayer method is characterized by lower cost, simpler operation, and easier osteogenic differentiation. The embryoid bodies (EBs) method is characterized by lower time consumption. Neuroscience Modeling the Functional Network for Spatial Navigation in the Human Brain Fengxiang Zhang*1, Chenghui Zhang*1, Yi Pu2, Xiang-Zhen Kong1,3 1Department of Psychology and Behavioral Sciences, Zhejiang University, 2Department of Neuroscience, Max Planck Institute for Empirical Aesthetics, 3Department of Psychiatry of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine This paper presents an integrative approach to investigating the functional network for spatial navigation in the human brain. This approach incorporates a large-scale neuroimaging meta-analytic database, resting-state functional magnetic resonance imaging, and network modeling and graph-theoretical techniques. Developmental Biology Generating Kidney Organoids in Suspension from Induced Pluripotent Stem Cells Langping Gao*1,2, Yue Wang*3, Gang Wang4, Hangdi Wu2, Qingtao Yan1, Jingjing Wang1, Fei Liu1, Haidong Fu1, Wei Li5, Lidan Hu1, Jianhua Mao1,2 1Department of Nephrology, The Children's Hospital, Zhejiang University School of Medicine, 2Zhejiang University School of Medicine, 3Hubei Normal University, 4National Clinical Research Center of Kidney Diseases, Jinling Hospital, Nanjing University School of Medicine, 5Clinical Laboratory, The Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health This protocol presents a comprehensive and efficient method for producing kidney organoids from induced pluripotent stem cells (iPSCs) using suspension culture conditions. The primary emphasis of this study lies in the determination of the initial cell density and the WNT agonist concentration, thereby benefiting investigators interested in kidney organoid research. Neuroscience Ultrasound-Guided Needle Release Combined with Corticosteroid Injection for the Treatment of Supinator Syndrome Zeng Zeng1,2, Cong-Xian Chen1 1Cancer Center, Department of Ultrasound Medicine, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, 2Zhejiang University School of Medicine The deep branch of the radial nerve can easily be compressed at the arcade of Frohse due to its anatomical features. Ultrasound-guided needle release combined with corticosteroid injection is an effective and safe treatment for deep branch radial nerve adhesion. Medicine Development and Evaluation of a Rat Model of Full-Thickness Cartilage Defects Haiyan Zhang*1, Ronghua Bao*2, Jiaan Xu3, Yanzhi Ge4, Zuxiang Chen1, Mengqiang Fan1, Guangping Yu5, Li Zhou1, Le Guo3, Letian Shan1, Hangxing Bao1 1The First Affiliated Hospital, Zhejiang Chinese Medical University, 2Fuyang Orthopaedics and Traumatology Affiliated Hospital, Zhejiang Chinese Medical University, 3College of Pharmaceutical Science, Zhejiang Chinese Medical University, 4Affiliated Zhejiang Hospital, Zhejiang University School of Medicine, 5Xianju County Hospital of Chinese Medicine This protocol establishes a full-thickness cartilage defects (FTCD) model by drilling holes in the femoral trochlear groove of rats and measuring the subsequent pain behavior and histopathological changes. Biochemistry Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles Xinghui Song1, Hao Shen2, Yanwei Li3, Yueting Xing3, Jiajia Wang3, Chun Guo3, Yingying Huang3, Jin Chen2 1Central Laboratory of Women’s Hospital, Zhejiang University School of Medicine, 2School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, 3Core Facilities, Zhejiang University School of Medicine This protocol provides a rapid and size-specific isolation method for small extracellular vesicles by optimizing the size of the air spray nozzle, sheath fluid pressure, sample flow pressure, voltage, gain, and triggering threshold parameters. Bioengineering Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde Yuelong Liang*1,2, Zhengze Huang*1, Yi Zhang*3,4, Yi Hong3,4, Qijiang Mao1,2,5, Xu Feng1 1Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Zhejiang Province Medical Research Center of Minimally Invasive Diagnosis and Treatment of Abdominal Diseases, 3Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Department of Orthopedic Surgery of the Second Affiliated Hospital, Zhejiang University School of Medicine, 4Zhejiang University-University of Edinburgh Institute, Zhejiang University School of Medicine, Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, 5Zhejiang Provincial Key Laboratory of Laparoscopic Technology The protocol presented here shows the synthesis of a strong adhesive hydrogel gelatin o-nitrosobenzaldehyde (gelatin-NB). Gelatin-NB has rapid and efficient tissue adhesion ability, which can form a strong physical barrier to protect wound surfaces, so it is expected to be applied to the field of injury repair biotechnology. Cancer Research An R-Based Landscape Validation of a Competing Risk Model Haiping Lin*1,3, Hongjuan Zheng*2, Chenyang Ge3, Lixian Ling2, Ruihua Yin2, Qinghua Wang2, Xia Zhang2, Shishi Zhou2, Xiayun Jin2, Xifeng Xu2, Jianfei Fu2 1Department of Hepatobiliary Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 2Department of Medical Oncology, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 3Department of Colorectal Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine The present protocol describes codes in R for evaluating the discrimination and calibration abilities of a competing risk model, as well as codes for the internal and external validation of it. Cancer Research Establishing a Competing Risk Regression Nomogram Model for Survival Data Lunpo Wu1,2, Chenyang Ge3, Hongjuan Zheng4, Haiping Lin5, Wei Fu6, Jianfei Fu4 1Department of Gastroenterology, Second Affiliated Hospital of Zhejiang University School of Medicine, 2Institute of Gastroenterology, Zhejiang University, 3Department of Colorectal Surgery, Afiliated Jinhua Hospital, Zhejiang University School of Medicine, 4Department of Medical Oncology, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 5Department of Hepatobiliary Pancreatic Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 6Division of Oncology, Johns Hopkins University School of Medicine Presented here is a protocol to build nomograms based on the Cox proportional hazards regression model and competing risk regression model. The competing method is a more rational method to apply when competing events are present in the survival analysis. Developmental Biology Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development Yong Wang*1, Ding Zhang*2, Fang Zhou2, Meijun Zhou2, Qiujie Li1, Jingyu Chen3, Jun Yang1,2 1Department of Physiology and Department of Cardiology of the Second Affiliated Hospital, Zhejiang University School of Medicine, 2Department of Cell Biology, State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, 3Wuxi Lung Transplant Center, Wuxi People's Hospital affiliated to Nanjing Medical University Presented here is a protocol for whole-mount in situ RNA hybridization analysis in zebrafish and tube formation assay in patient-derived induced pluripotent stem cell-derived endothelial cells to study the role of endoglin in vascular formation. Biology Light Spot-Based Assay for Analysis of Drosophila Larval Phototaxis Yixuan Sun1, Peipei Zhou1, Qianhui Zhao1, Zhefeng Gong1 1Department of Neurobiology, Zhejiang University This protocol introduces a light-spot assay to investigate Drosophila larval phototactic behavior. In this assay, a light spot is generated as light stimulation, and the process of larval light avoidance is recorded by an infrared light-based imaging system. Medicine A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts John Harry Caufield1,2, David A. Liem1,2,3, Anders O. Garlid1,2, Yijiang Zhou4, Karol Watson1,3, Alex A. T. Bui1,5,6,7, Wei Wang1,7,8,9, Peipei Ping1,2,3,7,8 1The NIH BD2K Center of Excellence in Biomedical Computing, University of California, Los Angeles, 2Department of Physiology, University of California, Los Angeles, 3Department of Medicine/Cardiology, University of California, Los Angeles, 4Department of Cardiology, First Affiliated Hospital, Zhejiang University School of Medicine, 5Department of Radiological Sciences, University of California, Los Angeles, 6Department of Bioengineering, University of California, Los Angeles, 7Scalable Analytics Institute (ScAi), University of California, Los Angeles, 8Department of Bioinformatics, University of California, Los Angeles, 9Department of Computer Science, University of California, Los Angeles We present a protocol and associated metadata template for the extraction of text describing biomedical concepts in clinical case reports. The structured text values produced through this protocol can support deep analysis of thousands of clinical narratives. Medicine Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes Jing Gao*1, Xiaolu Shi*2, Hong He1, Juhong Zhang1, Ding Lin3, Guosheng Fu1, Dongwu Lai1 1Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Experimental Research Center, China Academy of Chinese Medical Sciences, 3Department of Cardiology, The Third Hospital of Hangzhou City Intracellular calcium recycling plays a critical role in regulation of systolic and diastolic function in cardiomyocytes. Here, we describe a protocol to evaluate sarcoplasmic reticulum Ca2+ reserve and diastolic calcium removal function in cardiomyocytes by a calcium imaging system.