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Chinese PLA general hospital

20 ARTICLES PUBLISHED IN JoVE

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Biology

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
Li Wang 1, Graeme K. Carnegie 1
1Department of Pharmacology, University of Illinois at Chicago

Flow cytometric analysis of Bimolecular Fluorescence Complementation provides a high throughput quantitative method to study protein-protein interaction. This methodology can be applied to mapping protein binding sites and for screening factors that regulate protein-protein interaction.

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Biology

Massively Parallel Reporter Assays in Cultured Mammalian Cells
Alexandre Melnikov 1, Xiaolan Zhang 1, Peter Rogov 1, Li Wang 1, Tarjei S. Mikkelsen 1
1Broad Institute

The genetic reporter assay is a well-established and powerful tool for dissecting the relationship between DNA sequences and their gene regulatory activities. Coupling candidate regulatory elements to reporter genes that carry identifying sequence tags enables massive parallelization of these assays.

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Bioengineering

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
Yunfeng Chen *1, Baoyu Liu *2, Lining Ju *3, Jinsung Hong *1, Qinghua Ji 4,5, Wei Chen 6, Cheng Zhu 1
1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 3Charles Perkins Centre, The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University

We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte.

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Developmental Biology

Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5
Li Wang 1, Ziqing Liu 1, Chaoying Yin 1, Yang Zhou 1, Jiandong Liu 1, Li Qian 1
1Department of Pathology and Laboratory Medicine, McAllister Heart Institute, University of North Carolina, Chapel Hill

We describe here a protocol for the generation of iCMs using retrovirus-mediated delivery of Gata4, Tbx5 and Mef2c in a polycistronic construct. This protocol yields a relatively homogeneous population of reprogrammed cells with improved efficiency and quality and is valuable for future studies of iCM reprogramming.

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Immunology and Infection

Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells
Arash Nanbakhsh 1, Brad Best 2, Matthew Riese 3, Sridhar Rao 4, Li Wang 5, Jeffrey Medin 6, Monica S. Thakar 6, Subramaniam Malarkannan 1,5,6,7
1Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, The Blood Center of Wisconsin, 2Vector Core Lab, Blood Research Institute, The Blood Center of Wisconsin, 3Laboratory of Lymphocyte Biology, Blood Research Institute, The Blood Center of Wisconsin, 4Laboratory of Stem Cell Transcriptional Regulation, Blood Research Institute, The Blood Center of Wisconsin, 5Department of Microbiology and Immunology, The Medical College of Wisconsin, 6Department of Pediatrics, The Medical College of Wisconsin, 7Department of Medicine, The Medical College of Wisconsin

The goal of this study was to formulate technologies that allow for successful gene transduction in primary natural killer (NK) cells. The dextran-mediated lentiviral transduction of human or mouse primary NK cells results in higher gene expression efficiencies. This method of gene transduction will vastly improve NK cell genetic manipulation.

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Bioengineering

Microhoneycomb Monoliths Prepared by the Unidirectional Freeze-drying of Cellulose Nanofiber Based Sols: Method and Extensions
Zheng-Ze Pan 1,2, Hirotomo Nishihara 3, Wei Lv 1, Cong Wang 1,2, Yi Luo 1,2, Liubing Dong 1,2, Houfu Song 1,4, Wenjie Zhang 2, Feiyu Kang 1,2,4, Takashi Kyotani 3, Quan-Hong Yang 1,4,5
1Engineering Laboratory for Functionalized Carbon Materials and Shenzhen Key Laboratory for Graphene-based Materials, Graduate School at Shenzhen, Tsinghua University, 2School of Materials Science and Engineering, Tsinghua University, 3Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 4Tsinghua-Berkeley Shenzhen Institute (TBSI), Tsinghua University, 5School of Chemical Engineering and Technology, Tianjin University

Here, we present a general protocol to prepare a variety of microhoneycomb monoliths (MHMs) in which fluid can pass through with an extremely low pressure drop. MHMs obtained are expected to be used as filters, catalyst supports, flow-type electrodes, sensors and scaffolds for biomaterials.

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Neuroscience

Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats
Dandan Li *1,2, Hóngyi Zhào *2,3, Wei Wei 2, Nan Liu 2, Yonghua Dr. Huang 2
1Department of Neurology, Second Hospital of Shanxi Medical University, 2Department of Neurology, PLA Army General Hospital, 3Department of Neurology, NO 261 Hospital of PLA

We present a protocol to induce and detect CMHs caused by LPS injection in Sprague-Dawley rats, which may be utilized in future research investigations on the pathogenesis of CMHs.

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Neuroscience

Intrathecal Delivery of Antisense Oligonucleotides in the Rat Central Nervous System
Yi Chen *1, Curt Mazur *2, Yi Luo *1, Linhong Sun 1, Mingdi Zhang 1, Alex McCampbell 1, Giulio Srubek Tomassy 1
1Biogen, Inc., 2Ionis Pharmaceuticals

Here, we describe a method for delivering drugs to the rat central nervous system by implanting a catheter into the lumbar intrathecal space of the spine. We focus on the delivery of antisense oligonucleotides, though this method is suitable for delivery of other therapeutic modalities as well.

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Medicine

Traditional Trail Making Test Modified into Brand-new Assessment Tools: Digital and Walking Trail Making Test
Wei Wei *1, Hóngyi Zhào *1,2, Yu Liu 2, Yonghua Huang 1
1Department of Neurology, The Seventh Medical Center of PLA Army General Hospital, 2Department of Neurology, NO 984 Hospital of PLA

Here, we present a protocol to show how to perform two types of cognitive assessment tools derived from the paper-pencil version of the Trail Making Test.

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Bioengineering

Using Multilayered Hydrogel Bioink in Three-Dimensional Bioprinting for Homogeneous Cell Distribution
Nan Chen 1,2, Kai Zhu 1,2, Shiqiang Yan 3,4, Junmin Li 1,2, Tianyi Pan 4, Mieradilijiang Abudupataer 1,2, Fazle Alam 4, Xiaoning Sun 1,2, Li Wang 3,4, Chunsheng Wang 1,2
1Department of Cardiac Surgery and Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 2Shanghai Institute of Cardiovascular Diseases, 3Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, 4Institutes of Biomedical Sciences and Department of Systems Biology for Medicine, Shanghai Medical College, Fudan University

Here, we developed a novel multilayered modified strategy for liquid-like bioinks (gelatin methacryloyl with low viscosity) to prevent the sedimentation of encapsulated cells.

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Medicine

Digital Handwriting Analysis of Characters in Chinese Patients with Mild Cognitive Impairment
Hóngyi Zhào *1,2, Yanhai Zhang *3, Cuiqiao Xia 1, Yu Liu 2, Zhiyi Li 2, Yonghua Huang 1
1Department of Neurology, Chinese PLA General Hospital, 2Department of Neurology, NO 984 Hospital of PLA, 3Department of Cadre Ward, Xijing 986 Hospital, Fourth Military Medical University

This study proposed a digital handwriting analysis of characters in individuals with mild cognitive impairment to find more information than is revealed by traditional pencil–paper handwriting analysis.

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Bioengineering

Microfluidic Model to Mimic Initial Event of Neovascularization
Ping Zhao 1, Xing Zhang 1, Xiao Liu 1, Li Wang 4, Haoran Su 1, Liyi Wang 1, Dongrui Zhang 1, Xiaoyan Deng 3, Yubo Fan 1,2
1Beijing Advanced Innovation Centre for Biomedical Engineering, Key Laboratory for Biomechanics and Mechanobiology of Chinese Education Ministry, School of Biological Science and Medical Engineering, Beihang University, 2School of Engineering Medicine, Beihang University, 3Artificial Intelligence Key Laboratory of Sichuan Province, School of Automation and Information Engineering, Sichuan University of Science and Engineering, 4Beijing Research Center of Urban System Engineering

Here, we provide a microfluidic chip and an automatically controlled, highly efficient circulation microfluidic system that recapitulates the initial microenvironment of neovascularization, allowing endothelial cells (ECs) to be stimulated by high luminal shear stress, physiological level of transendothelial flow, and various vascular endothelial growth factor (VEGF) distribution simultaneously.

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Medicine

Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis
Li Huang 1, Wei Chen 1, Jiancong Chen 1, Dong Chen 1, Kunsong Zhang 1, Jianpeng Cai 1, Hong Peng 1, Chensong Huang 1, Guangyan Zeng 1, Mingjian Ma 1, Jiahua Liang 1, Borui Xu 1, Xiaoyu Yin 1, Jiaming Lai 1, Lijian Liang 1
1Department of Pancreatobiliary Surgery, The First Affiliated Hospital, Sun Yat-sen University

This protocol presents Indocyanine Green-guided video-assisted retroperitoneal debridement (ICG-guided VARD) for treating severe acute necrotizing pancreatitis.

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Genetics

Pre-Implantation Genetic Testing for Aneuploidy on a Semiconductor Based Next-Generation Sequencing Platform
Chengming Xu *1, Riqing Wei *2, Hui Lin 1, Leiyu Deng 1, Li Wang 3, Deyang Li 4, Honghui Den 5, Wensong Qin 1, Ping Wen 1, Ying Liu 1, Yingsong Wu 2, Qiang Ma 2, Jinliang Duan 1
1Centre for Women, Children, and Reproduction, Guangxi Key Laboratory of Metabolic Diseases Research, 2Department of Biopharmaceutics, School of Laboratory Medicine and Biotechnology, Southern Medical University, 3Department of Obstetrics and Gynecology, Chinese PLA General Hospital, 4Clinical Laboratory, Northern Theater Air Force Hospital, 5Guangzhou Darui Reproduction Technology Co., Ltd.

The protocol presents the overall in-lab procedures required in pre-implantation genetic testing for aneuploidy on a semiconductor-based next-generation sequencing platform. Here we present the detailed steps of whole genome amplification, DNA fragment selection, library construction, template preparation, and sequencing working flow with representative results.

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Cancer Research

Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells
Dongbo Xu 1, Li Wang 1, Kyle Wieczorek 1, Yanqing Wang 2, Xiaojing Zhang 2, David W. Goodrich 2, Qiang Li 1,2
1Department of Urology, Roswell Park Comprehensive Cancer Center, 2Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center

This protocol describes the process of the generation and characterization of mouse urothelial organoids harboring deletions in genes of interest. The methods include harvesting mouse urothelial cells, ex vivo transduction with adenovirus driving Cre expression with a CMV promoter, and in vitro as well as in vivo characterization.

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Behavior

Physical Activity Measurement in Children Accepting Table Tennis Training
Xin Zhang *1, Cuiqiao Xia *2, Xiuli Zhao 3, Yu Liu 3, Hóngyi Zhào 2,3, Yonghua Huang 2
1Department of Sport, Inner Mongolia Medical University, 2Department of Neurology, The Seventh Medical Center of PLA General Hospital, 3Department of Neurology, NO 984 Hospital of PLA

This study proposes an accelerometer-based method to objectively measure physical activity (PA) and leisure time physical activity (LTPA) in Chinese children accepting table tennis training in clubs.

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Biology

A Calcium Phosphate-Induced Mouse Abdominal Aortic Aneurysm Model
Siting Zhang *1, Zeyu Cai *1, Xu Zhang 1, Tianfeng Ma 2, Wei Kong 1
1Department of Physiology and Pathophysiology, School of Basic Medical Sciences; Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Peking University, 2Department of Vascular and Endovascular Surgery, the First Medical Centre, Chinese PLA General Hospital

This protocol describes a calcium phosphate-induced abdominal aortic aneurysm (AAA) mouse model to study the pathological features and molecular mechanisms of AAAs.

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Medicine

The Miniature Pig: A Large Animal Model for Cochlear Implant Research
Xiaojun Ji *1,2,3,4, Yi Luo *1,2,3,4, Weiwei Guo 1,2,3,4, Fei Ji 1,2,3,4, Shuolong Yuan 1,2,3,4, Liangwei Xu 1,2,3,4, Wei Chen *1,2,3,4
1College of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, 2National Clinical Research Center for Otolaryngologic Diseases, 3Key Lab of Hearing Science, Ministry of Education, 4Beijing Key Lab of Hearing Impairment for Prevention and Treatment

Miniature pigs (mini-pigs) are an ideal large animal model for research into cochlear implants. Cochlear implantation surgery in mini-pigs can be utilized to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches in a living system similar to human beings.

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Medicine

A Model of Long-Term Ventricular Fibrillation in Isolated Rat Hearts
Xiaoyi He 1,2, Libing Li 1, Wenhong Xu 1, Shengli Jiang 1,2
1Cardiovascular Surgery, First Medical Center, Chinese PLA General Hospital, 2Medical School of Chinese PLA

This protocol presents a model of long-term ventricular fibrillation in rat hearts induced by continuous stimulation with low-voltage alternating current. This model has a high success rate, is stable, reliable, and reproducible, has a low impact on cardiac function, and causes only mild myocardial injury.

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Medicine

Purification and Characterization of Extracellular Vesicles from Human Adipose-Derived Mesenchymal Stem Cells
Yuting Wang 1,2, Yan Han 1, Yudi Han 1
1Department of Plastic and Reconstructive Surgery, the First Medical Centre, Chinese PLA General Hospital, 2School of Medicine, Nankai University

This protocol describes all procedures, from culturing human adipose-derived mesenchymal stem cells (ADSCs) and collecting supernatant to extracting extracellular vesicles (EVs) using ultracentrifugation.

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