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Zhejiang University School of Medicine

44 ARTICLES PUBLISHED IN JoVE

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Biology

Human Pancreatic Islet Isolation: Part I: Digestion and Collection of Pancreatic Tissue
Meirigeng Qi 1, Barbara Barbaro 1, Shusen Wang 1, Yong Wang 1, Mike Hansen 1, Jose Oberholzer 1
1Department of Surgery, University of Illinois, Chicago

Achieving high quality and appropriate quantity of human islets is one of the prominent prerequisites for successful islet transplantation. In this video, we describe step by step the procedures for human pancreatic islet isolation (part I: digestion and collection of pancreatic tissue) using a modified automated method.

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Biology

Human Pancreatic Islet Isolation: Part II: Purification and Culture of Human Islets
Meirigeng Qi 1, Barbara Barbaro 1, Shusen Wang 1, Yong Wang 1, Mike Hansen 1, Jose Oberholzer 1
1Department of Surgery, University of Illinois, Chicago

Achieving high quality and appropriate quantity of human islets is one of the prominent prerequisites for successful islet transplantation. In this video, we describe step by step the procedures for human pancreatic islet isolation (part II: purification and culture of human islets) using a modified automated method.

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Biology

A Multi-Parametric Islet Perifusion System within a Microfluidic Perifusion Device
Adeola F. Adewola 1, Yong Wang 1, Tricia Harvat 1, David T. Eddington 2, Dongyoung Lee 1, Jose Oberholzer 1,2
1Department of Surgery, University of Illinois, Chicago, 2Department of Bioengineering, University of Illinois, Chicago

A microfluidic islet perifusion device was developed for the assessment of dynamic insulin secretion of multiple islets and simultaneous fluorescence imaging of calcium influx and mitochondrial potential changes.

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Immunology and Infection

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques
Hong He 1, Amy N. Courtney 1, Eric Wieder 2, K. Jagannadha Sastry 1
1Department of Immunology, MD Anderson Cancer Center - University of Texas, 2Department of Medicine, University of Miami

We demonstrate the utility of multicolor flow cytometry for detailed phenotypic and functional characterization of total as well as memory subsets of CD4+ and CD8+ T cells in rhesus macaques, the ideal model for HIV/AIDS vaccine studies.

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Bioengineering

Quantitative and Temporal Control of Oxygen Microenvironment at the Single Islet Level
Joe Fu-Jiou Lo 1, Yong Wang 2,3, Zidong Li 1, Zhengtuo Zhao 1, Di Hu 1, David T. Eddington 3, Jose Oberholzer 2,3
1Department of Mechanical Engineering, University of Michigan-Dearborn, 2Department of Surgery/Transplant, University of Illinois at Chicago, 3Department of Bioengineering, University of Illinois at Chicago

Microfluidic oxygen control confers more than just convenience and speed over hypoxic chambers for biological experiments. Especially when implemented via diffusion through a membrane, microfluidic oxygen can provide simultaneous liquid and gas phase modulations at the microscale-level. This technique enables dynamic multi-parametric experiments critical for studying islet pathophysiology.

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Biology

Fluorescence Imaging with One-nanometer Accuracy (FIONA)
Yong Wang *1,2, En Cai *1,2, Janet Sheung 1,2, Sang Hak Lee 1,2, Kai Wen Teng 2,3, Paul R. Selvin 1,2,3
1Department of Physics, University of Illinois at Urbana-Champaign, 2Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, 3Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign

Single fluorophores can be localized with nanometer precision using FIONA. Here a summary of the FIONA technique is reported, and how to carry out FIONA experiments is described.

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Chemistry

Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion
Siauw H. Ng 1, Yu Shi 1, Nicole E. Heshka 1, Yi Zhang 1, Edward Little 1
1CanmetENERGY, Natural Resources Canada

This paper presents an experimental method to produce biofuels and biochemicals from canola oil mixed with a fossil-based feed in the presence of a catalyst at mild temperatures. Gaseous, liquid, and solid products from a reaction unit are quantified and characterized. Conversion and individual product yields are calculated and reported.

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Engineering

Micro/Nano-scale Strain Distribution Measurement from Sampling Moiré Fringes
Qinghua Wang 1, Shien Ri 1, Hiroshi Tsuda 1
1Research Institute for Measurement and Analytical Instrumentation, National Institute of Advanced Industrial Science and Technology (AIST)

A sampling moiré technique featuring 2-pixel and multi-pixel sampling methods for high-accuracy strain distribution measurements at the micro/nano-scale is presented here.

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Medicine

Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes
Jing Gao *1, Xiaolu Shi *2, Hong He 1, Juhong Zhang 1, Ding Lin 3, Guosheng Fu 1, Dongwu Lai 1
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.

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Bioengineering

Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications
Taylor B. Dorsey 1,2,3, Alexander Grath 1,2, Cancan Xu 4, Yi Hong 4, Guohao Dai 1,2,3
1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 3Department of Bioengineering, Northeastern University, 4Department of Bioengineering, University of Texas at Arlington

In this method, we use photopolymerization and click chemistry techniques to create protein or peptide patterns on the surface of polyethylene glycol (PEG) hydrogels, providing immobilized bioactive signals for the study of cellular responses in vitro.

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Behavior

Use of a Video Scoring Anchor for Rapid Serial Assessment of Social Communication in Toddlers
Natasha Marrus 1, Stefanie Kennon-McGill 1, Brooke Harris 2, Yi Zhang 1, Anne L. Glowinski 1, John N. Constantino 1
1Department of Psychiatry, Washington University School of Medicine, 2Veteran's Administration Palo Alto Health Care System

Here we present a protocol for administration of the video-referenced rating of reciprocal social behavior (vrRSB) using a paper or online version. This survey quantifies RSB, a prerequisite for social competence, in toddlers through "video-referenced" items whereby a subject is compared to a reference child shown in a three-minute video.

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JoVE Core

Measuring the Carotid to Femoral Pulse Wave Velocity (Cf-PWV) to Evaluate Arterial Stiffness
Hongwei Ji 1, Jing Xiong 1, Shikai Yu 1, Chen Chi 1, Bin Bai 1, Jiadela Teliewubai 1, Yuyan Lu 1, Yi Zhang 1, Yawei Xu 1
1Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine

This protocol describes a method to standardize the measurements of carotid-femoral pulse wave velocity to evaluate arterial stiffness.

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Medicine

A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish
Xiao Ma *1,2,3, Yonghe Ding *2,3, Yong Wang 2,3,4, Xiaolei Xu 1,2,3
1Clinical and Translational Sciences Track, Mayo Clinic Graduate School of Biomedical Sciences, 2Department of Biochemistry and Molecular Biology, Mayo Clinic, 3Division of Cardiovascular Diseases, Mayo Clinic, 4Institute of Life Science, Beijing University of Chinese Medicine

A method to generate a doxorubicin-induced cardiomyopathy model in adult zebrafish (Danio rerio) is described here. Two alternative ways of intraperitoneal injection are presented and conditions to reduce variations among different experimental groups are discussed.

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Education

A Clinical Trial Assessing the Safety, Efficacy, and Delivery of Olive-Oil-Based Three-Chamber Bags for Parenteral Nutrition
Zhenyi Jia 1, Jun Yang 1, Huanlong Qin 1,2
1Department of General Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 2Department of General Surgery, Shanghai Tenth People's Hospital Affiliated to Tongji University

Here, we present a protocol for a study comparing the efficacy, safety, and delivery of olive-oil-based 3CB and soybean-oil-based CoB formulations in adults requiring parenteral nutrition. The results revealed that olive-oil-based 3CBs is non-inferior and well tolerated compared to soybean formulations.

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Medicine

A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts
John Harry Caufield 1,2, David A. Liem 1,2,3, Anders O. Garlid 1,2, Yijiang Zhou 4, Karol Watson 1,3, Alex A. T. Bui 1,5,6,7, Wei Wang 1,7,8,9, Peipei Ping 1,2,3,7,8
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.

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Cancer Research

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad
Gan-Lin Zhang 1, Yi Zhang 1, Ke-Xin Cao 1, Xiao-Min Wang 1
1Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University

Here, we present a protocol to implant breast cancer cells into the mammary fat pad in a simple, less invasive, and easy-to-handle way, and this mouse orthotopic breast cancer model with a proper mammary fat pad environment can be used to investigate various aspects of cancer.

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Chemistry

Reactive Inkjet Printing and Propulsion Analysis of Silk-based Self-propelled Micro-stirrers
David A. Gregory 1, Piyush Kumar 1, Ana Jimenez-Franco 1, Yi Zhang 1, Yu Zhang  2, Stephen J. Ebbens 1, Xiubo Zhao 1
1Department of Chemical and Biological Engineering, University of Sheffield, 2School of Engineering, University of Liverpool

This protocol demonstrates the ability to utilize reactive inkjet printing to print self-motile biocompatible and environmentally friendly micro-stirrers for use in biomedical and environmental applications.

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Medicine

Transvenous Embolization of Carotid Cavernous Fistula through Inferior Petrosal Sinus with Detachable Coils and Ethylene Vinyl Alcohol Copolymer
Jun Gu *1, Min Yan *1, Weijian Fan 1, Wenchao Liu 1, Ming Wang 1, Shu Wan 2
1Department of Neurosurgery, 1st affiliated Hospital, Zhejiang University School of Medicine, 2Brain Center, Zhejiang Hospital

The purpose of the article is to present our experience in endovascular treatment via the inferior petrosal sinus of the carotid cavernous fistula with detachable coils and ethylene-vinyl alcohol copolymer.

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Genetics

In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions
Lina Yu *1, Wenxiu He *1, Jie Xie *1, Rui Guo *1, Juan Ni 2, Xia Zhang 3, Quishi Xu 1, Caifeng Wang 1, Qiuling Yue 1, Fangfang Li 1, Mengcheng Luo 4, Bo Sun 3, Lan Ye 1, Ke Zheng 1
1State Key Laboratory of Reproductive Medicine, Nanjing Medical University, 2The Affiliated Hospital of Hangzhou Normal University, 3School of Life Science and Technology, ShanghaiTech University, 4Department of Tissue and Embryology, School of Basic Medical Sciences, Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan University

Presented here are protocols for in vitro biochemical assays using biotin labels that may be widely applicable for studying protein-nucleic acid interactions.

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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 Zhou 2, Meijun Zhou 2, Qiujie Li 1, Jingyu Chen 3, Jun Yang 1,2
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.

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Cancer Research

Establishing a Competing Risk Regression Nomogram Model for Survival Data
Lunpo Wu 1,2, Chenyang Ge 3, Hongjuan Zheng 4, Haiping Lin 5, Wei Fu 6, Jianfei Fu 4
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.

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Chemistry

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
Yi Zhang 1, Kanako Kurosawa 1, Daisuke Nishiura 2, Mika Tei 1, Mikiko Tsudome 3
1SUGAR Program, X-star, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2Center for Mathematical Science and Advanced Technology, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 3Deep-Sea Nanoscience Research Group, Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

The overall goal of the protocol is to prepare over one million ordered, uniform, stable, and biocompatible femtoliter droplets on a 1 cm2 planar substrate that can be used for cell-free protein synthesis.

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JoVE Core

High-Temperature and High-Pressure In situ Magic Angle Spinning Nuclear Magnetic Resonance Spectroscopy
Nicholas R. Jaegers 1, Wenda Hu 2, Yong Wang 1, Jian Zhi Hu 1
1Pacific Northwest National Laboratory, 2Washington State University

The molecular structures and dynamics of solids, liquids, gases, and mixtures are of critical interest to diverse scientific fields. High-temperature, high-pressure in situ MAS NMR enables detection of the chemical environment of constituents in mixed phase systems under tightly controlled chemical environments.

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Developmental Biology

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila
Tongchao Li 1, Liqun Luo 1
1Department of Biology, Howard Hughes Medical Institute, Department of Biology, Stanford University

This protocol describes the dissection procedure, culture condition, and live imaging of an antennae-brain explant system for the study of the olfactory circuit assembly.

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Education

Improved Renal Denervation Mitigated Hypertension Induced by Angiotensin II Infusion
Ming Wang *1,2, Shuyi Zhang *1,2, Wanling Han 1,2, Maoqing Ye 1,2, Xinkai Qu 1,2, Wenzheng Han 1,2
1Department of Cardiology, Huadong Hospital Affiliated to Fudan University, 2Shanghai Key Laboratory of Clinical Geriatric Medicine

Here, we present a protocol for renal sympathetic denervation (RDN) in mice with hypertension induced by Angiotensin II infusion. The procedure is repeatable, convenient and allows to study the regulatory mechanisms of RDN on hypertension and cardiac hypertrophy.

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Cancer Research

An R-Based Landscape Validation of a Competing Risk Model
Haiping Lin *1,3, Hongjuan Zheng *2, Chenyang Ge 3, Lixian Ling 2, Ruihua Yin 2, Qinghua Wang 2, Xia Zhang 2, Shishi Zhou 2, Xiayun Jin 2, Xifeng Xu 2, Jianfei Fu 2
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.

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Biochemistry

Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles
Xinghui Song 1, Hao Shen 2, Yanwei Li 3, Yueting Xing 3, Jiajia Wang 3, Chun Guo 3, Yingying Huang 3, Jin Chen 2
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.

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Medicine

Development and Evaluation of a Rat Model of Full-Thickness Cartilage Defects
Haiyan Zhang *1, Ronghua Bao *2, Jiaan Xu 3, Yanzhi Ge 4, Zuxiang Chen 1, Mengqiang Fan 1, Guangping Yu 5, Li Zhou 1, Le Guo 3, Letian Shan 1, Hangxing Bao 1
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.

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Neuroscience

Real-Time Dynamic Collection of Hippocampal Extracellular Fluid from Conscious Rats Using a Microdialysis System
Xiaobo Wang 1,2, Na Xie 3, Yi Zhang 4, Xianli Meng 1,2,3, Ya Hou 3, Sanyin Zhang 1,2
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 4School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The protocol here provides a detailed real-time dynamic sampling of extracellular fluid from the hippocampus of awake rats using a microdialysis system.

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Medicine

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique
Ya Hou 1, Jinrong Bai 1,2, Yi Zhang 3, Xianli Meng 1,2,4, Sanyin Zhang 4, Xiaobo Wang 2
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 4Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine

The present protocol describes a simple and efficient method for the real-time and dynamic collection of rat heart blood using the microdialysis technique.

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Methods And Protocols For Live Imaging In Development
Tongchao Li 1,2,3
1Department of Neurobiology and Department of Neurosurgery of Second Affiliated Hospital, Zhejiang University School of Medicine, 2Liangzhu Laboratory, MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, 3NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University

Methods And Protocols For Live Imaging In Development

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JoVE Core

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules
Qinyun Du 1, Qingxiu He 2, Fengming Zhang 3, Jiao Mi 3, Yuying Li 3, Shaohui Wang 1, Yi Zhang 1
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 3Tibet Rhodiola Pharmaceutical Holding Co. Ltd

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

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Bioengineering

Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde
Yuelong Liang *1,2, Zhengze Huang *1, Yi Zhang *3,4, Yi Hong 3,4, Qijiang Mao 1,2,5, Xu Feng 1
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.

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Medicine

Voltage-Dependent Potassium Current Recording on H9c2 Cardiomyocytes via the Whole-Cell Patch-Clamp Technique
Hong Jiang *1, Yating Zhang *1, Ya Hou 1, Lijun Li 2, Sanyin Zhang 3,4, Yi Zhang 5, Xianli Meng 1,3,4, Xiaobo Wang 4
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Chengdu Techman Instrument Co., Ltd., 3Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 4Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 5School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes an efficient method for the real-time and dynamic acquisition of voltage-gated potassium (Kv) channel currents in H9c2 cardiomyocytes using the whole-cell patch-clamp technique.

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Neuroscience

Modeling the Functional Network for Spatial Navigation in the Human Brain
Fengxiang Zhang *1, Chenghui Zhang *1, Yi Pu 2, Xiang-Zhen Kong 1,3
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.

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Neuroscience

Ultrasound-Guided Needle Release Combined with Corticosteroid Injection for the Treatment of Supinator Syndrome
Zeng Zeng 1,2, Cong-Xian Chen 1
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.

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Medicine

Unilateral Biportal Endoscopy for the Treatment of Lumbar Disc Herniation
Zihao Yu 1,2,3,4, Chenyi Ye 1,2,3,4, Mhd Alaa Alhendi 5, Hua Zhang 1,2,3,4
1Department of Orthopedic Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, 2Orthopedics Research Institute of Zhejiang University, 3Key Laboratory of Motor System Disease Research and Precision Therapy of Zhejiang Province, 4Clinical Research Center of Motor System Disease of Zhejiang Province, 5Zhejiang University School of Medicine

Presented here is a step-by-step surgical protocol of unilateral biportal endoscopy (UBE), a minimally invasive approach for treating lumbar disc herniation. By using two ports, surgical accuracy and flexibility are enhanced. This article provides a comprehensive description of UBE and highlights the practical application of this approach in clinical practice.

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Developmental Biology

Generating Kidney Organoids in Suspension from Induced Pluripotent Stem Cells
Langping Gao *1,2, Yue Wang *3, Gang Wang 4, Hangdi Wu 2, Qingtao Yan 1, Jingjing Wang 1, Fei Liu 1, Haidong Fu 1, Wei Li 5, Lidan Hu 1, Jianhua Mao 1,2
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.

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Biology

Comparison of Two Representative Methods for Differentiation of Human Induced Pluripotent Stem Cells into Mesenchymal Stromal Cells
Fangqin Wang 1, Langping Gao 2,3, Xudong Fu 4, Qintao Yan 2, Lidan Hu 1,2, Jianhua Mao 2,3
1National Clinical Research Center for Child Health, The Children's Hospital, Zhejiang University School of Medicine, 2Department 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.

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Medicine

Bilateral Renal Ischemia-Reperfusion Model for Acute Kidney Injury in Mice
Junni Wang *1, Huanhuan Zhu *1, Jin Miao 1, Weiqiang Lin 2, Fei Han 1
1Kidney Disease Center, the First Affiliated Hospital, Zhejiang University School of Medicine, 2International Institute of Medicine, The Fourth Affiliated Hospital, Zhejiang University School of Medicine

The ischemia-reperfusion injury (IRI) model can be used at different stages of acute kidney injury (AKI) development, especially during AKI to chronic kidney disease (CKD) progression. Here, we describe the procedure for IRI model development in mice via a trans-abdominal approach, clamping renal pedicles via a vascular clip to induce ischemic injury.

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Bioengineering

Cell Patterning Using Magnetic-Archimedes Strategy
Xuhao Zhou 1,2,3, Miribani Maitusong 1,2,3, Tanchen Ren 1,2,3, Yan Wu 1,2,3
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.

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Medicine

Combination of Anatomical Plates and Rotator Cuff Reinforcement for Minimally Invasive Treatment of Greater Tuberosity Fractures of the Humerus
Xiao Lu *1, Xiaodi Zou *2, Ahmad Alhaskawi 3, Yanzhao Dong 3, Sohaib Hasan Abdullah Ezzi 4, Vishnu Goutham Kota 5, Mohamed Hasan Abdulla Hasan Abdulla 5, Sahar Ahmed Abdalbary 6, Hui Lu 3,7
1Department of Orthopedics, Sanmen People's Hospital, 2Department of Orthopedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, 3Department of Orthopedics, The First Affiliated Hospital, Zhejiang University, 4Department of Orthopedics, Third Xiangya Hospital, Central South University, 5Zhejiang University School of Medicine, 6Department of Orthopedic Physical Therapy, Faculty of Physical Therapy, Nahda University in Beni Suef, 7Alibaba-Zhejiang University Joint Research Center of Future Digital Healthcare, Zhejiang University

Here, we present a protocol to utilize anatomical plates alongside rotator cuff reinforcement in a minimally invasive surgery to efficiently treat humerus greater tuberosity fractures, promising quicker healing and improved stability.

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Medicine

A Minimally Invasive, Visualized Method for Nasojejunal Tube Placement
Kun Li *1, Jin Huang *2, Ahmad Alhaskawi 3, Bin Wang 2, Nianbin Ma 4, Chengjun Yao 5, Yanzhao Dong 3, QingFang Zhao 6, Xiaodi Zou 7, Hui Lu 3
1Department of Critical Care Medicine, The First Affiliated Hospital, Zhejiang University School of Medicine, 2Department of Emergency Medicine, People's Hospital of Anji County, 3Department of Orthopaedics, The First Affiliated Hospital, Zhejiang University School of Medicine, 4Department of Critical Care Medicine, People's Hospital of Anji County, 5Zhejiang University School of Medicine, 6Department of Plastic Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, 7Department of Orthopedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Xinhua Hospital of Zhejiang Province

This study introduces a bedside visualized method to improve nasojejunal tube placement in intensive care unit patients, enhancing efficiency and reducing patient discomfort.

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Medicine

A Modified Technique for Inducing Polycystic Ovary Syndrome in Mice
Bokang Zhou *1, Fei Guo *1, Yuhang Long 1, Junsen She 2,3, Ling Gao 1,2,4,5, Hefeng Huang 1,2,3,4,5
1Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Fudan University, 2Women's Hospital, Key Laboratory of Reproductive Genetics (Ministry of Education), Zhejiang University School of Medicine, 3International Institutes of Medicine, the Fourth Affiliated Hospital, Zhejiang University School of Medicine, 4Research Units of Embryo Original Diseases, Chinese Academy of Medical Sciences (No. 2019RU056), 5State Key Laboratory of Cardiology

This protocol introduces a technique for inducing polycystic ovary syndrome (PCOS) models in mice through controlled letrozole release using mini-pumps. Under adequate anesthesia, the mini-pump was implanted subcutaneously, and PCOS was successfully induced in the mice after a certain period of the mini-pump release.

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