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Guangzhou Medical University

20 ARTICLES PUBLISHED IN JoVE

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JoVE Core

Fabrication and Operation of a Nano-Optical Conveyor Belt
Jason Ryan 1, Yuxin Zheng 1, Paul Hansen 1, Lambertus Hesselink 2
1Electrical Engineering, Stanford University, 2Applied Physics, Stanford University

The scalability and resolution of conventional optical manipulation techniques are limited by diffraction. We circumvent the diffraction limit and describe a method of optically transporting nanoparticles across a chip using a gold surface patterned with a path of closely spaced C-shaped plasmonic resonators.

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Developmental Biology

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro
Zhaofeng Jia 1,2, Yujie Liang 3, Bin Ma 4,5, Xiao Xu 2,6, Jianyi Xiong 2, Li Duan 2, Daping Wang 1,2
1Guangzhou Medical University, 2Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopeadic Engineering, Department of Orthopedics, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), 3Department of Chemistry, The Chinese University of Hong Kong, 4School of Biomedical Engineering, Shanghai Jiao Tong University, 5Renji Hospital Clinical Stem Cell Research Center, Shanghai Jiao Tong University School of Medicine, 6Shantou University Medical College

We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.

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Neuroscience

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization
Xiao Xu 1, Yinwei You 1,2, Long Zhang 1
1Department of Entomology, China Agricultural University, 2Bio-tech Research Center, Shandong Academy of Agricultural Sciences

This paper describes a detailed and highly effective RNA in situ hybridization protocol particularly for low-level expressed Odorant Receptor (OR) genes, as well as other genes, in insect antennae using digoxigenin (DIG)-labeled or biotin-labeled probes.

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Developmental Biology

Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
Cui Lin 1, Dongxing Zhu 2, Greg Markby 1, Brendan M. Corcoran 3, Colin Farquharson 1, Vicky E. Macrae 1
1Developmental Biology, The Roslin Institute and R(D)SVS, University of Edinburgh, 2Guangzhou Institute of Cardiovascular Disease, The Second Affiliated Hospital, School of Basic Medical Sciences, Guangzhou Medical University, 3Clinical Sciences and R(D)SVS, University of Edinburgh

This protocol describes the isolation, culture, and calcification of rat-derived valve interstitial cells, a highly physiological in vitro model of calcific aortic valve disease (CAVD). Exploitation of this rat model facilitates CAVD research in exploring the cell and molecular mechanisms that underlie this complex pathological process.

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Biology

Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model
Zhaofeng Jia 1,2,3, Yujie Liang 4,5, Xingfu Li 2,3, Xiao Xu 1,2,3, Jianyi Xiong 2,3, Daping Wang 1,2,3, Li Duan 2,3
1Postgraduate institution, Guangzhou Medical University, 2Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, 3Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopaedic Engineering, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), 4Department of Chemistry, Chinese University of Hong Kong, 5Shenzhen Kangning Hospital, Shenzhen Mental Health Center

This article presents a simple and economic protocol for the straightforward isolation and purification of mesenchymal stem cells from New Zealand white rabbit synovial fluid.

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Immunology and Infection

A Silver Nanoparticle Method for Ameliorating Biliary Atresia Syndrome in Mice
Ming Fu 1, Zefeng Lin 2, Huiting Lin 2, Yanlu Tong 2, Hezhen Wang 2, Hongjiao Chen 2, Yan Chen 2, Ruizhong Zhang 2
1First Affiliated Hospital of Jinan University, 2Department of Pediatric Surgery, Guangzhou Women & Children's Medical Center, Guangzhou Medical University

This article describes in detail a method based on silver nanoparticles for ameliorating biliary atresia syndrome in an experimental biliary atresia mouse model. A solid understanding of the reagent preparation process and the neonatal mouse injection technique will help familiarize researchers with the method used in neonatal mouse model studies.

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Genetics

Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
Yan Yan *1, Chi-Chih Chang *1, Morten T. Venø 1,2, Colin R. Mothershead 1, Junyi Su 1, Jørgen Kjems 1,2
1The Interdisciplinary Nanoscience Centre, Aarhus University, 2Department of Molecular Biology and Genetics, Aarhus University

This protocol demonstrates how to purify extracellular microRNAs from cell culture media for small RNA library construction and next generation sequencing. Various quality control checkpoints are described to allow readers to understand what to expect when working with low input samples like exRNAs.

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Neuroscience

Diffusion Tensor Magnetic Resonance Imaging in Chronic Spinal Cord Compression
Weipeng Zheng *1, Xiuhang Ruan *2, Xinhua Wei 3, Fangtian Xu 4, Yuanping Huang 1, Ning Wang 1, Haoyi Chen 5, YingJie Liang 6, Wende Xiao 6, Xin Jiang 1, Shifeng Wen 6
1Department of Orthopedics, Guangzhou First People's Hospital, Guangzhou Medical University, 2Department of Radiology, Guangzhou First People's Hospital, Guangzhou Medical University, 3Department of Radiology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, 4Department of Orthopedics, The First Affiliated Hospital of Gannan Medical University, 5Department of Orthopedics, Guangzhou Chest Hospital, 6Department of Orthopedics, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou Medical University

Here, we present a protocol for the application of diffusion tensor imaging parameters to evaluate spinal cord compression.

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Biochemistry

Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers
Yi-Xin Huo 1,2, Bo Zheng 1, Ning Wang 1, Yunpeng Yang 1, Xinxin Liang 1, Xiaoyan Ma 1
1Key Laboratory of Molecular Medicine and Biotherapy, School of Life Sciences, Beijing Institute of Technology, 2UCLA Institute for Technology Advancement (Suzhou)

This study presents an alternative strategy to the conventional toxic analog-based method in identifying amino acid overproducers by using rare-codon-rich markers to achieve accuracy, sensitivity, and high-throughput simultaneously.

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Biology

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice
Qiancheng Wang 1, Kaitong Chen 1, Hairuo Lin 1, Mingyuan He 2, Xiaoxia Huang 1, Hailin Zhu 1, Yulin Liao 1
1Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 2The Third Clinical Medical College, Guangzhou Medical University

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

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Medicine

The Lower Body Positive Pressure Treadmill for Knee Osteoarthritis Rehabilitation
Junjie Liang *1,2, Yuanyuan Guo *1,2, Yuxin Zheng 1,2, Shijuan Lang 1, Hongxin Chen 1,2, Yaoyao You 1,2, Bryan O’Young 3,4, Haining Ou *1,2, Qiang Lin *1,2
1Department of Rehabilitation, The Fifth Affiliated Hospital of Guangzhou Medical University, 2Experiment Education Model Center of Rehabilitation Medicine, Guangzhou Medical University, 3Department of Physical Medicine and Rehabilitation, Geisinger Health System, 4Department of Rehabilitation Medicine, New York University School of Medicine

Here, based on a clinician’s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

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Biology

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae
Chuanxian Wei *1, Yan Yan *1, Xiaoli Miao 1, Renjie Jiao 1,2,3
1Sino-French Hoffmann Institute, School of Basic Medical Sciences, Guangzhou Medical University, 2The Second Affiliated Hospital of Guangzhou Medical University, 3The State Key Laboratory of Respiratory Disease, Guangzhou Medical University

Presented here are detailed methods for the dissection and lipid droplet staining of oenocytes in Drosophila larvae using BODIPY 493/503, a lipid droplet-specific fluorescent dye.

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Behavior

Quantitative Static and Dynamic Assessment of Balance Control in Stroke Patients
Qiang Lin *1,2,3,4, Yuxin Zheng *1,2,3,4, Peixi Lian 3,4, Yuanyuan Guo 1,3,4, Haojie Huang 5, Zhiwei Luo 1,2, Wanying Li 1,2, Hongxin Chen 1,2,3,4, Sufen Li 1, Shijuan Lang 1,2,4, Yumei Chen 6, Haining Ou *1,2,3,4, Junjie Liang *1,2,3,4
1Department of Rehabilitation, The Fifth Affiliated Hospital of Guangzhou Medical University, 2The Rehabilitation Medicine Lab of Guangzhou Medical University, 3Experiment Education Model Center of Rehabilitation Medicine, Guangzhou Medical University, 4Department of Rehabilitation Therapy, Guangzhou Medical University, 5College of Physical Education and Sports, Beijing Normal University, 6Department of Rehabilitation, Nanfang Hospital, Southern Medical University

Presented here is a quantitative, clinical balance assessment method suitable for stroke patients with balance disorders.

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Behavior

Effects of Surgical Masks on Cardiopulmonary Function in Healthy Subjects
Mei Li *1,2,4, Haining Ou *1,2,3,4, Qinyi Li 1,2,4, Junjie Liang 1,2,4, Weihao Liao 1,2,4, Shijuan Lang 1,2,4, Hongxin Chen 1,2,4, Yafei Wang 5, Kun Tang 1,2,4, Qiang Lin *1,2,3,4, Yuxin Zheng *1,2,4
1Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, 2The Rehabilitation Medicine Lab of Guangzhou Medical University, 3Guangzhou Key Laboratory of Enhanced Recovery after Abdominal Surgery, The Fifth Affiliated Hospital of Guangzhou Medical University, 4Department of Rehabilitation Therapy, Guangzhou Medical University, 5Department of Rehabilitation Medicine, The Sixth Affiliated Hospital of Sun Yat-sun University

We study the effect of surgical masks on cardiopulmonary function based on a cardiopulmonary exercise test (CPET). This study shows that surgical masks reduce cardiopulmonary exercise capacity and ventilation in healthy young subjects and wearing masks might affect aerobic exercise capacity more in female subjects than in male subjects.

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Developmental Biology

Using Chicken Embryo as a Powerful Tool in Assessment of Developmental Cardiotoxicities
Qixiao Jiang 1, Xiaohui Xu 1, Jamie C. DeWitt 2, Yuxin Zheng 1
1School of Public Health, Qingdao University, 2Department of Pharmacology & Toxicology, Brody School of Medicine, East Carolina University

Chicken embryos, as a classical developmental model, are used in our lab to assess developmental cardiotoxicities following exposure to various environmental contaminants. Exposure methods and morphological/functional assessment methods established are described in this manuscript.

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Biology

Large-Scale Preparation of Synovial Fluid Mesenchymal Stem Cell-Derived Exosomes by 3D Bioreactor Culture
Li Duan 1,2, Xingfu Li 1,2, Xiao Xu 1,2, Limei Xu 1,2, Daping Wang 1,2, Kan Ouyang 1,2, Yujie Liang 1,3
1Department of Orthopedics, The First affiliated hospital of Shenzhen University, Shenzhen Second People's Hospital, 2Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, Shenzhen Second People's Hospital, 3Department of Child and Adolescent Psychiatry, Shenzhen Kangning Hospital, Shenzhen Mental Health Center, Shenzhen Key Laboratory for Psychological Healthcare & Shenzhen Institute of Mental Health

Here, we present a protocol to produce a large number of GMP-grade exosomes from synovial fluid mesenchymal stem cells using a 3D bioreactor.

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Neuroscience

Motor Dual-Tasks for Gait Analysis and Evaluation in Post-Stroke Patients
Haining Ou *1,2,3,4, Shijuan Lang *1,2,3, Yuxin Zheng 1,2,3, Dongqing Huang 2, Sheng Gao 1, Meifeng Zheng 1,2, Biyi Zhao 2, Zulipiya Yiming 2, Yaxian Qiu 1,2, Qiang Lin *1,2,3,4, Junjie Liang *1,2,3
1Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, 2Department of Rehabilitation Medicine, Guangzhou Medical University, 3Rehabilitation Medicine Lab, Guangzhou Medical University, 4Guangzhou Key Laboratory of Enhanced Recovery after Abdominal Surgery, The Fifth Affiliated Hospital of Guangzhou Medical University

This paper presents a protocol specifically for dual motor task gait analysis in stroke patients with motor control deficits.

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Medicine

A Neonatal BALB/c Mouse Model of Necrotizing Enterocolitis
Yan Tian *1, Junyu Huang *1, Ming Fu 1, Qiuming He 1, Jiale Chen 1, Yan Chen 1, Ruizhong Zhang 1, Wei Zhong 1
1Provincial Key Laboratory of Research in Structure Birth Defect Disease and Department of Pediatric Surgery, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight infants, with high mortality and unclear pathogenesis. The cause of NEC may be related to inflammatory immune regulatory system abnormalities. An NEC animal model is an indispensable tool for NEC disease immune research. NEC animal models usually use C57BL/6J neonatal mice; BALB/c neonatal mice are rarely used. Related studies have shown that when mice are infected, Th2 cell differentiation is predominant in BALB/c mice compared to C57BL/6J mice. Studies have suggested that the occurrence and development of NEC are associated with an increase in T helper type 2 (Th2) cells and are generally accompanied by infection. Therefore, this study used neonatal BALB/c mice to induce an NEC model with similar clinical characteristics and intestinal pathological changes as those observed in children with NEC. Further study is warranted to determine whether this animal model could be used to study Th2 cell responses in NEC.

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Medicine

Generation and Characterization of Right Ventricular Myocardial Infarction Induced by Permanent Ligation of the Right Coronary Artery in Mice
Ruoxi Liao 1, Mingyuan He 2, Donghong Hu 2, Chengzheng Gong 1, Huiying Du 1, Hairuo Lin 2, Huijun Sun 3
1The First Clinical College of Dalian Medical University, 2Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 3College of Pharmacy, Dalian Medical University

There are several differences between the right and left ventricles. However, the pathophysiology of right ventricular infarction (RVI) has not been clarified. In the present protocol, a reproducible method for RVI mouse model generation is introduced, which may provide a means to explain the mechanism of RVI.

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Neuroscience

Dual-Task Stroop Paradigm for Detecting Cognitive Deficits in High-Functioning Stroke Patients
Songjun Lin 1,2, Qiang Lin 3,4,5, Biyi Zhao 3,4,6, Yongchun Jiang 3,4, Wanqi Zhuang 3,4, Delong Chen 3,4, Yajie Zhang 3,4, Aijia Chen 3,4, Qianrong Zhang 7, Yuxin Zheng 3,4, Jianjun Wang 1, Fangqiu Xu 8, Xi Qin 3,4,9, Yefeng Cai 10
1Department of Neurology and Psychology, the Fourth Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen Traditional Chinese Medicine Hospital, 2Guangzhou University of Chinese Medicine, 3Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, 4Department of Rehabilitation Medicine, Guangzhou Medical University, 5Key Laboratory of Biological Targeting Diagnosis, Therapy and Rehabilitation of Guangdong Higher Education Institutes, The Fifth Affiliated Hospital of Guangzhou Medical University, 6School of Traditional Chinese Medicine, Jinan University, 7Health College of Guangdong Pharmaceutical University, 8Department of Clinical Medicine, Guangzhou Medical University, 9Department of Rehabilitation Medicine, Panzhihua Central Hospital, 10Department of Neurology and Psychology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine

Clinical assessment scales are notsensitive enough to cognitive dysfunction in high-functioning stroke patients. The dual-task paradigm presents advantages and potential in the assessment and cognitive training of cognitive dysfunction. The study here proposes a dual-task Stroop paradigm to identify cognitive dysfunction in high-functioning stroke patients.

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