JoVE Logo

Sign In

Capital Medical University

65 ARTICLES PUBLISHED IN JoVE

image

Biology

Drosophila Pupal Abdomen Immunohistochemistry
Wei Wang 1, John H. Yoder 1
1Department of Biological Sciences, University of Alabama

Antibody staining of the Drosophila pupae can enhance genetic analyses of adult abdominal developmental genetics. We present our protocol for dissection, fixation and antibody staining of staged Drosophila pupal abdomen.

image

Medicine

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window
Lifeng Zhang *1, Andree Lapierre *1, Brittany Roy 1, Maili Lim 1, Jennifer Zhu 1, Wei Wang 1, Stephen B. Sampson 1, Kyuson Yun 1, Bonnie Lyons 1, Yun Li 1, Da-Ting Lin 1
1The Jackson Laboratory

By combining a polished and reinforced thin-skull (PoRTS) cranial window and glioblastoma (GBM) cell injection, we can observe glioma initiation and growth from injected GBM cells in the brain of a live mouse longitudinally.

image

Neuroscience

Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons
Yun Li 1, Brittany D. Roy 1, Wei Wang 1, Lifeng Zhang 1, Stephen B. Sampson 1, Da-Ting Lin 1
1The Jackson Laboratory

By tagging the extracellular domains of membrane receptors with superecliptic pHluorin, and by imaging these fusion receptors in cultured mouse neurons, we can directly visualize individual vesicular insertion events of the receptors to the plasma membrane. This technique will be instrumental in elucidating the molecular mechanisms governing receptor insertion to the plasma membrane.

image

Medicine

RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma
Hongwei Wang 1, Mindy Xiao-Ming Wang 1, Nan Su 1, Li-chong Wang 1, Xingyong Wu 1, Son Bui 1, Allissa Nielsen 1, Hong-Thuy Vo 1, Nina Nguyen 1, Yuling Luo 1, Xiao-Jun Ma 1
1Advanced Cell Diagnostics, Inc.

The presence of high-risk HPV in head and neck tumor tissue is associated with favorable outcomes. The recently developed RNA in situ hybridization technique called RNAscope allows direct visualization of HPV E6/E7 mRNA in FFPE tissue sections.

image

Immunology and Infection

Generation of Lymphocytic Microparticles and Detection of their Proapoptotic Effect on Airway Epithelial Cells
Chun Yang *1, Wei Xiong *2, Qian Qiu 2, Houda Tahiri 1, Carmen Gagnon 1, Guoxiang Liu 2, Pierre Hardy 1
1Departments of Pediatrics and Pharmacology, University of Montréal, 2Department of Pulmonology, Chongqing Southwest Hospital, Third Military Medical University

Cell membrane–shed microparticles (MPs) are active biological vesicles that can be isolated and their pathophysiological effects investigated in various models. Here we describe a method for generating MPs derived from T lymphocytes (LMPs) and for demonstrating their proapoptotic effect on airway epithelial cells.

image

Chemistry

Facile and Efficient Preparation of Tri-component Fluorescent Glycopolymers via RAFT-controlled Polymerization
Wei Wang 1, John M. Lester 1, Anthony E. Amorosa 2, Deborah L. Chance 3, Valeri V. Mossine 1, Thomas P. Mawhinney 1,4
1Department of Biochemistry, University of Missouri, 2Department of Biological Sciences, University of Missouri, 3Department of Molecular Microbiology & Immunology, University of Missouri, 4Department of Child Health, University of Missouri

An efficient, three-step synthesis of RAFT-based fluorescent glycopolymers, consisting of glycomonomer preparation, copolymerization, and post-modification, is demonstrated. This protocol can be used to prepare RAFT-based statistical glycopolymers with desired structures.

image

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.

image

Bioengineering

Expanding Nanopatterned Substrates Using Stitch Technique for Nanotopographical Modulation of Cell Behavior
Kai Wang 1, Kam W. Leong 2, Yong Yang 1
1Department of Chemical and Biomedical Engineering, West Virginia University, 2Department of Biomedical Engineering, Columbia University

A protocol for producing a large area of nanopatterned substrate from small nanopatterned molds for study of nanotopographical modulation of cell behavior is presented.

image

Bioengineering

A Microfluidic System with Surface Patterning for Investigating Cavitation Bubble(s)–Cell Interaction and the Resultant Bioeffects at the Single-cell Level
Fenfang Li 1, Fang Yuan 2, Georgy Sankin 1, Chen Yang 1, Pei Zhong 1
1Mechanical Engineering and Materials Science, Duke University, 2Huacells Corp

A microfluidic chip was fabricated to produce pairs of gold dots for tandem bubble generation and fibronectin-coated islands for single-cell patterning nearby. The resultant flow field was characterized by particle image velocimetry and was employed to study various bioeffects, including cell membrane poration, membrane deformation, and intracellular calcium response.

image

Developmental Biology

Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells
Yueying Li 2, Yong Hai 1, Jiayu Chen 3, Tie Liu 1
1Department of Orthopedics, Beijing Chao-Yang Hospital, Capital Medical University, 2Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, 3Clinical and Translational Research Center of Shanghai First Maternity & Infant Hospital, School of Life Sciences and Technology, Tongji University

We present a protocol to generate a chondrogenic lineage from human peripheral blood (PB) via induced pluripotent stem cells (iPSCs) using an integration-free method, which includes embryoid body (EB) formation, fibroblastic cells expansion, and chondrogenic induction.

image

Engineering

New Application of an Atmospheric Pressure Plasma Jet as a Neuro-protective Agent Against Glucose Deprivation-induced Injury of SH-SY5Y Cells
Xu Yan *1,3,4,5, Zhaozhong Meng *2, Jiting Ouyang 2, Yajun Qiao 2, Fang Yuan 1,3,4,5
1Department of Pathophysiology, Beijing Neurosurgical Institute/Beijing Tiantan Hospital, Capital Medical University, 2School of Physics, Beijing Institute of Technology, 3Beijing Key Laboratory of Central Nervous System Injury, Beijing Neurosurgical Institute, Beijing Tiantan Hospital, Capital Medical University, 4Beijing Key Laboratory of Translational Medicine for Cerebrovascular Disease, Beijing Tiantan Hospital, Capital Medical University, 5China National Clinical Research Center for Neurological Diseases, Beijing Tiantan Hospital, Capital Medical University

A protocol for the neuroprotective application of low-dose atmospheric pressure plasma treatment on glucose deprivation-induced SH-SY5Y injuries.

image

Immunology and Infection

An Efficient and Simple Method to Establish NK and T Cell Lines from Patients with Chronic Active Epstein-Barr Virus Infection
Chongfeng Xu 1, Junhong Ai 2, Qingxun Zhang 1, Ting Li 3, Xiaorong Wu 1, Zhengde Xie 2, Ziyuan Duan 1
1Genetic Resources Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 2MOE Key Laboratory of Major Diseases in Children, National Key Discipline of Pediatrics, National Clinical Research Center for Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, 3State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences

A simple method for obtaining NK and T cell clones from CAEBV patients was developed with high efficiency, a small amount of peripheral blood, and a low-dose of IL-2.

image

Biology

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents
Feng Pan *2, Jie Zhang *3, Yuyan Liu 1, Liangsheng Lu 1, Xuefeng Qiu 4, Kangtai Lv 5, Qipeng Zhang 1
1State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, 2Department of Andrology, The Affiliated Obstetrics and Gynecology Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, 3School of Pharmaceutical Sciences, Nanjing Tech University, 4Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, 5Department of Ultrasound, Nanjing Qixia District Maternity and Child Health Care Hospital

Intracavernosal pressure recording (ICP) is an important method to evaluate the erectile function of experimental animals. Here, a detailed protocol is demonstrated for the recording procedure of ICP by catheterizing the crura penis and then electrically stimulating the cavernous nerves in rats.

image

Developmental Biology

Zika Virus Infection of Cultured Human Fetal Brain Neural Stem Cells for Immunocytochemical Analysis
Erica L. McGrath 1,2, Junling Gao 1, Ping Wu 1,3
1Department of Neuroscience and Cell Biology, University of Texas Medical Branch, 2Department of Neurology, Johns Hopkins University, 3Beijing Institute for Brain Disorders, Capital Medical University

This article details the methods that are used to expand human fetal brain neural stem cells in culture, as well as how to differentiate them into various neuronal subtypes and astrocytes, with an emphasis on the use of neural stem cells to study Zika virus infection.

image

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.

image

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.

image

Genetics

Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice
Jing-Ying Guo 1, Lu He 1, Teng-Fei Qu 1, Yu-Ying Liu 2, Ke Liu 1, Guo-Peng Wang 1, Shu-Sheng Gong 1
1Department of Otolaryngology-Head and Neck Surgery, Beijing Friendship Hospital, Capital Medical University, 2Department of Otolaryngology-Head and Neck Surgery, Shanghai First People's Hospital, Shanghai Jiao Tong University

Here we describe canalostomy procedure which allows local drug delivery into the inner ears of adult and neonatal mice through the semicircular canal with minimum damage to hearing and vestibular function. This method can be used to inoculate viral vectors, pharmaceuticals, and small molecules into the mouse inner ear.

image

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.

image

Medicine

A Pleural Effusion Model in Rats by Intratracheal Instillation of Polyacrylate/Nanosilica
Wen Cao 1, Xiaoli Zhu 2, Ziren Tang 3, Yuguo Song 2
1Department of Ultrasound Medicine, Beijing Chaoyang Hospital, Capital Medical University, 2Department of Occupational Medicine & Clinical Toxicology, Beijing Chaoyang Hospital, Capital Medical University, 3Department of Emergency, Beijing Chaoyang Hospital, Capital Medical University

Here, we present a protocol to construct a pleural effusion model in rats by intratracheal instillation of polyacrylate/nanosilica.

image

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.

image

Engineering

Nerve Stem Cell Differentiation by a One-step Cold Atmospheric Plasma Treatment In Vitro
Zilan Xiong *1, Shasha Zhao *2, Xu Yan 3
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.

image

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.

image

Medicine

Cloud-Based Phrase Mining and Analysis of User-Defined Phrase-Category Association in Biomedical Publications
Dibakar Sigdel *1,2, Vincent Kyi *1,2, Aiden Zhang *1, Shaun P. Setty 3, David A. Liem 1,2,4, Yu Shi 5, Xuan Wang 5, Jiaming Shen 5, Wei Wang 1,6,7, JiaWei Han 5, Peipei Ping 1,2,4,6
1The NIH BD2K Center of Excellence in Biomedical Computing, University of California, Los Angeles, 2Department of Physiology, University of California, Los Angeles, 3Department of Pediatric and Adult Congenital Heart Surgery, Miller Children's and Women's Hospital and Long Beach Memorial Hospital, 4Department of Medicine/Cardiology, University of California, Los Angeles, 5NIH BD2K Program Centers of Excellence for Big Data Computing -- KnowEng Center, Department of Computer Science, University of Illinois at Urbana-Champaign (UIUC), 6Scalable Analytics Institute (ScAi), University of California, Los Angeles, 7Department of Computer Science, University of California, Los Angeles

We present a protocol and associated programming code as well as metadata samples to support a cloud-based automated identification of phrases-category association representing unique concepts in user selected knowledge domain in biomedical literature. The phrase-category association quantified by this protocol can facilitate in depth analysis in the selected knowledge domain.

image

JoVE Core

Morphological and Functional Evaluation of Ribbon Synapses at Specific Frequency Regions of the Mouse Cochlea
Shu-Kui Yu *1, Zheng-De Du *1, Qing-Ling Song 1, Teng-Fei Qu 1, Yue Qi 1, Wei Xiong 1, Lu He 1, Wei Wei 2, Shu-Sheng Gong 1, Ke Liu 1
1Department of Otolaryngology-Head and Neck Surgery, Beijing Friendship Hospital, Capital Medical University, 2Department of Otology, Shengjing Hospital of China Medical University

This manuscript describes an experimental protocol for evaluating the morphological characteristics and functional status of ribbon synapses in normal mice. The present model is also suitable for noise-induced and age-related cochlear synaptopathy-restricted models. The correlative results of previous mouse studies are also discussed.

image

JoVE Journal

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure
Honghui Mao *1,2, Xiuchao Wang *1,3, Wen Chen 4, FengYu Liu 5, You Wan 5, Sanjue Hu 1, Junling Xing 1,6
1Department of Neurobiology, School of Basic Medicine, Fourth Military Medical University, 2Department of Toxicology, School of Public Health, ShanXi Medical University, 3Department of Psychology, Fourth Military Medical University, 4Department of Neurobiology, School of Basic Medical Sciences, Advanced Innovation Center for Human Brain Protection, Capital Medical University, 5Neuroscience Research Institute, Key Lab for Neuroscience, Ministry of Education/National Health Commission, Peking University, 6Department of Radiation Biology, Faculty of Preventive Medicine, Fourth Military Medical University

Single-fiber recording is an effective electrophysiological technique that is applicable to the central and peripheral nervous systems. Along with the preparation of intact DRG with the attached sciatic nerve, the mechanism of conduction failure is examined. Both protocols improve the understanding of the peripheral nervous system's relationship with pain.

image

Medicine

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach
Ali R. Ahmadi 1, Le Qi 1, Kenichi Iwasaki 1, Wei Wang 1, Russell N. Wesson 1, Andrew M. Cameron 1, Zhaoli Sun 1
1Department of Surgery, Johns Hopkins University School of Medicine

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

image

Environment

A Uniaxial Compression Experiment with CO2-Bearing Coal Using a Visualized and Constant-Volume Gas-Solid Coupling Test System
Weitao Hou 1, Hanpeng Wang 1, Wei Wang 1, Zhongzhong Liu 1, Qingchuan Li 1
1Research Center of Geotechnical and Structural Engineering, Shandong University

This protocol demonstrates how to prepare a briquette sample and conduct a uniaxial compression experiment with a briquette in different CO2 pressures using a visualized and constant-volume gas-solid coupling test system. It also aims to investigate changes in terms of coal’s physical and mechanical properties induced by CO2 adsorption.

image

Medicine

Three-Dimensional Printing Guide Template Assisted Percutaneous Vertebroplasty (PVP)
PeiLun Hu *1, JiSheng Lin *1, JunChuan Xu 1, Hai Meng 1, Nan Su 1, Yong Yang *1, Qi Fei 1
1Department of Orthopedics, Beijing Friendship Hospital, Capital Medical University

Herein, we present a three-dimensional printing guide template for percutaneous vertebraplasty. A patient with a T11 vertebral compression fracture was selected as a case study.

image

Medicine

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Di Liu *1, Xizhu Xu *2, Yuejin Li 2, Jie Zhang 1, Xiaoyu Zhang 1, Qihuan Li 1, Haifeng Hou 2, Dong Li 2, Wei Wang 1,2,3, Youxin Wang 1
1Beijing Key Laboratory of Clinical Epidemiology, School of Public Health, Capital Medical University, 2School of Public Health, Shandong First Medical University & Shandong Academy of Medical Sciences, 3School of Medical and Health Sciences, Edith Cowan University

Immunoglobulin G (IgG) N-glycan is characterized using hydrophilic interaction chromatography UPLC. In addition, the structure of IgG N-glycan is clearly separated. Presented here is an introduction to this experimental method so that it can be widely used in research settings.

image

Cancer Research

Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane
Moe Ishihara *1, Junhui Hu *1, Xiaoyu Zhang 2, YongHyeon Choi 3, Anthony Wong 4, Celine Cano-Ruiz 5, Rongwei Zhao 6, Ping Tan 7, Jonathan L. Tso 1, Lily Wu 1,8
1Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, 2Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, 3Department of Bioengineering, Hanyang University, 4Department of Ecology and Evolutionary Biology, University of California, Los Angeles, 5Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, 6School of Life Sciences, Beijing Normal University, 7Department of Urology, West China Hospital, Sichuan University, 8Department of Urology, David Geffen School of Medicine, University of California, Los Angeles

Metastatic clear cell renal cell carcinoma is a disease without a comprehensive animal model for thorough preclinical investigation. This protocol illustrates two novel animal models for the disease: the orthotopically implanted mouse model and the chicken chorioallantoic membrane model, both of which demonstrate lung metastasis resembling clinical cases.

image

JoVE Core

A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice
Jie Zhang 1,2, Fang Fang 1,3, Liang Li 1, Haoliang Huang 1, Hannah C. Webber 1, Yang Sun 1,4, Vinit B. Mahajan 1,4, Yang Hu 1
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.

image

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.

image

Behavior

A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice
Yi Xie 1, Sai-Yue Deng 1, Si-Miao Chen 1, Xue-Jiao Chen 1, Wen-Wen Lai 1, Li-Fang Huang 1, Li Ba 1, Wei Wang 1,2, Feng-Fei Ding 1,3
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.

image

Neuroscience

Detrusor Underactivity Model in Rats by Conus Medullaris Transection
Xin Zheng *1, Menghua Wu *1, Jian Song 2, Jimao Zhao 2
1Department of Urology, Beijing YouAn Hospital, Capital Medical University, 2Department of Urology, Beijing Friendship Hospital, Capital Medical University

We present a method for establishing a detrusor underactivity model by conus medullaris transection in rats. Detrusor underactivity was successfully stimulated in these animals. The model can be used for studying urinary tract function.

image

Neuroscience

Establishment of a Rat Model of Superior Sagittal-Sinus Occlusion via a Thread-Embolism Method
Weiyan Jiang *1, Congcong Jin *2, Weiming Xu *3, Yingxian Li 4, Yinghong Lin 5, Shengxiang Liang 6,7, Wei Wang 8
1Department of Clinical Laboratory, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 2Rehabilitation Department, Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine, 3Department of Neurosurgery, the Hospital of Changle, 4Pediatrics Ward 2, Shouguang Hospital of Traditional Chinese Medicine, 5Fujian University of Traditional Chinese Medicine, 6National-Local Joint Engineering Research Center of Rehabilitation Medicine Technology, Fujian University of Traditional Chinese Medicine, 7Rehabilitation Industry Institute, Fujian University of Traditional Chinese Medicine, 8Department of Neurosurgery, the First Affiliated Hospital of Wenzhou Medical University

Here, we establish a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis via a thread-embolization method, and the stability and reliability of the model were verified.

image

Medicine

Directed Induction of Retinal Organoids from Human Pluripotent Stem Cells
Xiao Zhang 1, Zi-Bing Jin 1
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Science Key Laboratory

Using a self-organizing method, we develop a protocol with the addition of COCO that could significantly increase the generation of photoreceptors.

image

Medicine

Modifications of the Langendorff Method for Simultaneous Isolation of Atrial and Ventricular Myocytes from Adult Mice
Kui Wu 1, Lin-Ling Li 2, Yu-Kun Li 1, Xiao-Dong Peng 1, Meng-Xia Zhang 1, Ke-Sen Liu 1, Xue-Si Wang 1, Jia-Xue Yang 1, Song-Nan Wen 1, Yan-Fei Ruan 1, Nian Liu 1, Rong Bai 1
1Department of Cardiology, Bejing Anzhen Hospital, Capital Medical University, 2Department of Cardiology, Bejing Chuiyangliu Hospital

This protocol describes modifications of the Langendorff method including the depth of aorta cannulation for simultaneous Isolation of atrial and ventricular myocytes from adult mice.

image

Cancer Research

Reconstruct Human Retinoblastoma In Vitro
Xiao Zhang 1, Zi-Bing Jin 1
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Science Key Laboratory

We describe a method for generating human retinoblastoma (RB) by introducing biallelic RB1 mutations in human embryonic stem cells (hESC). RB cell lines could also be successfully cultured using the isolated RB in a dish.

image

Neuroscience

Modified Spared Nerve Injury Surgery Model of Neuropathic Pain in Mice
Liangliang He *1,2, Wenxing Zhao *1, Lingyi Zhang 2, Maalveka Ilango 2,3, Na Zhao 2, Liqiang Yang 1, Zhonghui Guan 2
1Department of Pain Management, Xuanwu Hospital, Capital Medical University, 2Department of Anesthesia and Perioperative Care, University of California San Francisco, 3University of Washington

The modified surgery is a simplified method for mouse or rat spared nerve injury model that requires only one ligation and one cut to injure both common peroneal and sural nerves.

image

Medicine

Application of Optical Coherence Tomography to a Mouse Model of Retinopathy
Xiaoting Mai *1, Shaofen Huang *1, Wen Chen 1, Tsz Kin Ng 1,2,3, Haoyu Chen 1
1Joint Shantou International Eye Center, Shantou University and the Chinese University of Hong Kong, 2Shantou University Medical College, 3Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong

Here, we describe an in vivo imaging technique using optical coherence tomography to facilitate the diagnosis and quantitative measurement of retinopathy in mice.

image

Developmental Biology

Generation of Human Patient iPSC-derived Retinal Organoids to Model Retinitis Pigmentosa
Chao Ma *1, Kangxin Jin *1, Zi-Bing Jin 1
1Beijing Tongren Hospital, Beijing Institute of Ophthalmology, Beijing Ophthalmology & Visual Science Key Laboratory, Beijing Tongren Eye Center, Capital Medical University

In this protocol, retinitis pigmentosa patient induced pluripotent stem cell (iPSC)-derived 3D retinal organoids were generated. Those organoids successfully recapitulated some clinical phenotypes of the retinitis pigmentosa disease.

image

Biology

A Mouse Model of Pulmonary Fibrosis Induced by Nasal Bleomycin Nebulization
Dingyun Song 1,2, Yicong Chen 1,3, Xin Wang 1,4, Xueying Chen 1,2, Shuwei Gao 1,3, Wenxiu Xu 1,3, Shengnan Yang 5, Zai Wang 6, Liang Peng 7, Huaping Dai 1,2
1Department of Pulmonary and Critical Care Medicine, Center of Respiratory Medicine, China-Japan Friendship Hospital, National Clinical Research Center for Respiratory Diseases, Institute of Respiratory Medicine, Chinese Academy of Medical Sciences, 2Graduate School of Peking Union Medical College, Chinese Academy of Medical Science and Peking Union Medical College, 3Capital Medical University, 4Beijing University of Chinese Medicine, 5Tianjin Chest Hospital, 6Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, 7Beijing Key Laboratory for Immune-Mediated Inflammatory Diseases, Institute of Clinical Medical Sciences, China-Japan Friendship Hospital

Various animal models of pulmonary fibrosis have been established using bleomycin to clarify the pathogenesis of pulmonary fibrosis and find new drug targets. However, most pulmonary fibrosis models targeting lung tissue have uneven drug administration. Here, we propose a model of uniform pulmonary fibrosis induced by nasal bleomycin nebulization.

image

Biology

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
Jinxi Yuan 1, Jie Zhang 1, Yan Zhang 1, WuYun QiQiGe 1, Wei Liu 2, Shanchun Yan 1, Guirong Wang 2
1Key Laboratory of Sustainable Forest Ecosystem Management - Ministry of Education, Northeast Forestry University, 2Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences

This paper presents the step-by-step protocols for CRISPR/Cas9 mutagenesis of the Oriental fruit fly Bactrocera dorsalis. Detailed steps provided by this standardized protocol will serve as a useful guide for generating mutant flies for functional gene studies in B. dorsalis.

image

Immunology and Infection

Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis
Linlin Zhang 1,2, Mengjia Liu 1,2, Meng Zhang 1,2, Junhong Ai 1,2, Jiao Tian 1,2, Ran Wang 1,2, Zhengde Xie 1,2
1Beijing Key Laboratory of Pediatric Respiratory Infectious Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, 2Research Unit of Critical Infection in Children, 2019RU016, Chinese Academy of Medical Sciences

We describe a method combining immunomagnetic beads and fluorescence-activated cell sorting to isolate and analyze defined immune cell subpopulations of peripheral blood mononuclear cells (monocytes, CD4+ T cells, CD8+ T cells, B cells, and natural killer cells). Using this method, magnetic and fluorescently labeled cells can be purified and analyzed.

image

Cancer Research

Surgical Technique for Superior Cervical Ganglionectomy in a Murine Model
Qi Wang 1, Chun-Hao Chen 1, Hongbo Xu 2, Sylvie Deborde 1,3, Richard J. Wong 1,3
1Department of Surgery, Memorial Sloan Kettering Cancer Center, 2Department of Otolaryngology-Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, 3David M. Rubenstein Center for Pancreatic Cancer Research, Memorial Sloan Kettering Cancer Center

The present protocol describes a mouse model of the ablation of adrenergic innervation by identifying and resecting the superior cervical ganglion.

image

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.

image

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.

image

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.

image

Immunology and Infection

Detection of Polyfunctional T Cells in Children Vaccinated with Japanese Encephalitis Vaccine via the Flow Cytometry Technique
Linlin Zhang *1,2, Meng Zhang *1,2,3, Mengjia Liu 1,2, Junhong Ai 1,2, Jiao Tian 1,2, Huizheng Ge 4, Ran Wang 1,2, Zhengde Xie 1,2
1Beijing Key Laboratory of Pediatric Respiratory Infectious Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, 2Research Unit of Critical Infection in Children, 2019RU016, Chinese Academy of Medical Sciences, 3Department of Pediatric Rehabilitation, Beijing Boai Hospital, School of Rehabilitation Medicine, Capital Medical University, China Rehabilitation Research Center, 4Center of Excellence (Beijing), Becton Dickinson Medical Devices (Shanghai) Co Ltd

The present protocol combines ex vivo stimulation and flow cytometry to analyze polyfunctional T cell (TPF) profiles in peripheral blood mononuclear cells (PBMCs) within Japanese encephalitis virus (JEV)-vaccinated children. The detection method and flow cytometry color scheme of JEV-specific TPFs were tested to provide a reference for similar studies.

image

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.

image

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.

image

Neuroscience

A Procedure for Mouse Dorsal Root Ganglion Cryosectioning
Liangliang He *1,2, Wenxing Zhao *1, Lingyi Zhang 2,3, Maalveka Ilango 2,4, Na Zhao 2, Liqiang Yang 1, Zhonghui Guan 2
1Department of Pain Management, Xuanwu Hospital, Capital Medical University, 2Department of Anesthesia and Perioperative Care, University of California San Francisco, 3Department of Anesthesiology, Sun Yat-Sen University, 4University of Washington

Presented here is the development for consistently acquiring high-quality dorsal root ganglion cryostat sections.

image

Medicine

A 3D Digital Model for the Diagnosis and Treatment of Pulmonary Nodules
Jingwen Xue 1, Fangliang Xing 2, Yang Liu 3, Tengxiao Liang 4
1Beijing University of Chinese Medicine, 2Beijing Intelligent Entropy Science & Technology Co Ltd., 3Center for Integrated Chinese and Western Medicine, Beijing You An Hospital, Capital Medical University, 4Fever Outpatient Clinic, Dongzhimen Hospital of Beijing University of Chinese Medicine

The objective of this study is to develop a novel 3D digital model of pulmonary nodules that serves as a communication bridge between physicians and patients and is also a cutting-edge tool for pre-diagnosis and prognostic evaluation.

image

Medicine

A Three-Dimensional Digital Model for Early Diagnosis of Hepatic Fibrosis Based on Magnetic Resonance Elastography
Yang Liu 1, Tengxiao Liang 2, Fangliang Xing 3, Wei Hou 4, Xiaofei Shang 1, Xiuhui Li 1
1Center for Integrated Chinese and Western Medicine, Beijing You An Hospital, Capital Medical University, 2Fever Clinics, Dongzhimen Hospital of Beijing University of Chinese Medicine, 3Beijing Intelligent Entropy Science& Technology Co Ltd., 4First Department of The Liver Disease Center, Beijing You An Hospital, Capital Medical University

The objective of this study was to develop a novel three-dimensional digital model for the early diagnosis of hepatic fibrosis, which includes the stiffness of each voxel in the patient's liver and can thus, be used to calculate the distribution ratio of the patient's liver at different fibrosis stages.

image

Medicine

Three-Dimensional Reconstruction for the Whole Lung with Early Multiple Pulmonary Nodules
Junyi Shi 1, Fangliang Xing 2, Yang Liu 3, Tengxiao Liang 4
1Beijing University of Chinese Medicine, 2Beijing Intelligent Entropy Science & Technology Co Ltd., 3Center for Integrated Chinese and Western Medicine, Beijing You An Hospital, Capital Medical University, 4Fever Outpatient Clinic, Dongzhimen Hospital of Beijing University of Chinese Medicine

This study introduces a three-dimensional (3D) reconstruction method for the entire lung in patients with early multiple pulmonary nodules. It offers a comprehensive visualization of nodule distribution and their interplay with lung tissue, simplifying the assessment of diagnosis and prognosis for these patients.

image

A Rat Model of Compound Acne
Lin Chen *1,2, Qi Jin *1,2, Tian Xia *2, Xin Zhang 2, Ruiyi Cai 1, Xinyi Wu 2, Yue-Han Song 2, Guang-Zhong Zhang 1
1Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, 2School of Traditional Chinese Medicine, Beijing University of Chinese Medicine

This protocol outlines the procedure for inducing acne inflammation in rat skin with oleic acid and Cutibacterium acnes.

image

Medicine

Investigating the Effect of Different Types of Exercise on Upper Limb Functional Recovery in Patients with Right Hemisphere Damage Based on fNIRS
Yu Wei *1,2, JiaLi Chen *3, Rui Fang *1,2, JingYa Liu 1,2, MengChen Feng 1,2, HuiRong Du 2, MeiQing Wang 2, Reziya Abulihaiti 2, Hua Ling 2,4, FuBiao Huang 1,2
1Department of Occupational Therapy, China Rehabilitation Research Center, 2Faculty of Rehabilitation, Capital Medical University, 3College of Sports Health, Tianjin Institute of Physical Education, 4China Rehabilitation Science Institute

Here, we investigate the effect of functional occupational therapy combined with assisted active or passive motion on the upper limb function of patients with right hemisphere damage and explore the effect of functional near-infrared spectroscopy on brain function remodeling.

image

Medicine

Examination of Oral Candida Infection in Primary Sjögren's Syndrome Patients
Xiaoyan Wang 1,2,3, Yeqing Song 4, Xinzhe Lou 1, Bingqing Han 5,6,7, Ruiqing Wu 1, Jing Xie 8,9, Xiang Lin 8,9, Hao Wang 1
1Department of Stomatology, Beijing Tiantan Hospital, Capital Medical University, 2Beijing Laboratory of Oral Health, Capital Medical University, 3Department of Biochemistry and Molecular Biology, School of Basic Medicine, Capital Medical University, 4Central Laboratory, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, 5Clinical Diagnosis Laboratory of Beijing Tiantan Hospital, Capital Medical University, 6Key Laboratory for Quality Control of In Vitro Diagnostics, NMPA, 7Beijing Engineering Research Center of Immunological Reagents Clinical Research, 8School of Chinese Medicine, The University of Hong Kong, 9Department of Rheumatology and Clinical Immunology, Beijing Tiantan Hospital, Capital Medical University

Here, we present a protocol for the examination of oral Candida infection in patients with primary Sjögren's syndrome, which can be used for timely treatment and, thereafter, avoiding related complications.

image

Bioengineering

Effects of Extracorporeal Membrane Oxygenation on the Coagulation System
Tianlong Wang *1, Weidong Yan *2, Mingru Zhang 3, Jing Wang 1, Han Zhang 1, Gang Liu 1, Jian Wang 1, Bingyang Ji 1
1Department of Cardiopulmonary Bypass, Fuwai Hospital, National Center for Cardiovascular Disease, State Key Laboratory of Cardiovascular Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, 2Department of Anesthesiology, Beijing Chao-Yang Hospital, Capital Medical University, 3Department of Anesthesiology, Beijing Tongren Hospital, Capital Medical University

We present a protocol for establishing a long-term awake extracorporeal membrane oxygenation (ECMO) model in sheep. Special attention is given to the management and evaluation of the coagulation system during the ECMO model.

image

Immunology and Infection

Postoperative Ileus Murine Model
Dan Tian *1,2,3,4,5, Bihan Liu *1,2,3,4,5, Shiyang Huang 2,3,4,5,6, Wen Shi 2,3,4,5,6, Yue Tian 2,3,4,5,6, Yingchi Yang 1,2,3, Dong Zhang 7,8, Zhongtao Zhang 1,2,3, Fandi Bu 9
1General Surgery Department, Beijing Friendship Hospital, Capital Medical University, 2National Clinical Research Center for Digestive Diseases, 3State Key Lab of Digestive Health, Beijing Friendship Hospital, Capital Medical University, 4Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University, 5Beijing Laboratory of Oral Health, Capital Medical University School of Stomatology, 6Beijing Clinical Research Institute, 7Medical Research Center, Beijing Chaoyang Hospital, Capital Medical University, 8Beijing Institute of Brain Disorders, Capital Medical University, 9Department of Hernia and Abdominal Wall Surgery, Beijing Chaoyang Hospital, Capital Medical University

We describe a murine model of postoperative ileus generated via intestinal manipulation. Gastrointestinal transit function, pathologic changes, and immune cell activation were assessed 24 h after surgery.

image

Medicine

Nine-Grid Area Division Method: A New Ideal Bone Puncture Region for Percutaneous Vertebroplasty in Lumbar Spine
Jisheng Lin *1, Weiyang Zuo *1, Peng Wu *2, Xiaoyu Li 3, Hai Meng 1, Jinjun Li 1, Qi Fei 1
1Department of Orthopedics, Beijing Friendship hospital, Capital Medical University, 2Defense Innovation Institute, Chinese Academy of Military Science, 3Department of Emergency Medicine, Aerospace Center Hospital

Herein, we present a "Nine-grid Area Division Method" for percutaneous vertebroplasty. A patient with an L1 vertebral compression fracture was selected as a case study.

image

Biology

Mitochondrial Transfer Via Tunneling Nanotubes Between Mesenchymal Stem Cells and Retinal Pigment Epithelium In Vitro
Jing Yuan 1, Zi-Bing Jin 1
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University

Here, we present a protocol including mitochondrial tracing, direct co-culture procedures of mesenchymal stem cells (MSCs) and retinal pigment epithelial cells (ARPE19), as well as the methods for observing and statistically analyzing tunneling nanotubes (TNT) formation and mitochondrial transfer to characterize mitochondrial exchange via TNTs between MSCs and ARPE19 cells.

image

Neuroscience

Assembling Retinal Organoids with Microglia
Jia Xu *1, Si-Jian Yu *1, Zi-Bing Jin 1
1Beijing Institute of Ophthalmology, Beijing Tongren Hospital, Capital Medical University

Microglia are unique resident immune cells in the retina, playing crucial roles in various retinal degenerative diseases. Generating a co-culture model of retinal organoids with microglia can facilitate a better understanding of the pathogenesis and development progress of retinal diseases.

image

Biology

In Vitro Model for Studying Differentiation and Changes of Multi-Omics on Murine Airway Epithelial Cells Stimulated with Cigarette Smoke Extract
Muzhi Zhang 1, Liuyinuo Zhao 1, Kaixin Li 1, Ruoyang Zhang 2,3, Zhe Lv 1, Jie Liu 1, Ye Cui 1, Wei Wang 1, Sun Ying 1
1Department of Immunology, School of Basic Medical Sciences, Capital Medical University, 2Department of Pulmonary and Critical Care Medicine, Center of Respiratory Medicine, China-Japan Friendship Hospital, 3National Center for Respiratory Medicine

Here, we describe an in vitro model for isolating and differentiating murine airway epithelial cells, focusing on their acclimation to chronic cigarette smoke extract (CSE). The model could be utilized to comprehensively characterize the multi-omics impact of CSE, which possibly provides insights into the cellular responses under chronic smoke exposure.

image

Neuroscience

Longitudinal In Vivo Imaging of Retinal Cells with Tailored Adeno-Associated Virus Serotypes via Confocal Scanning Laser Ophthalmoscopy
Tianmin Ren 1, Xiaowei Fan 1, Kailai Chen 2, Jiashuo Liu 2, Jingxue Zhang 1,3,4, Ningli Wang 1,3,4
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Laboratory, 2Second Clinical Medical School, Capital Medical University, 3Beijing Institute of Brain Disorders, Collaborative Innovation Center for Brain Disorders, Capital Medical University, 4Beijing Key Laboratory of Fundamental Research on Biomechanics in Clinical Application, Capital Medical University

This protocol describes the use of adeno-associated virus (AAV) vectors for cell-specific labeling and in vivo imaging using a confocal scanning laser ophthalmoscope (CSLO). This method enables the investigation of different retinal cell types and their contributions to retinal function and disease.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved