The Chinese University of Hong Kong 25 articles published in JoVE Medicine Erosion Identification in Metacarpophalangeal Joints in Rheumatoid Arthritis using High-Resolution Peripheral Quantitative Computed Tomography Yousif Al-Khoury1,2, Stephanie Finzel3, Camille Figueiredo4, Andrew J. Burghardt5, Kathryn S. Stok6, Lai-Shan Tam7, Isaac Cheng7, Justin J. Tse1, Sarah L. Manske1 1Department of Radiology, McCaig Institute for Bone and Joint Health, University of Calgary, 2Department of Biomedical Engineering, University of Calgary, 3University Medical Center Freiburg, 4Department of Rheumatology, University of São Paulo, 5Department of Radiology and Biomedical Imaging, University of California, San Francisco, 6Department of Biomedical Engineering, University of Melbourne, 7Department of Medicine and Therapeutics, The Chinese University of Hong Kong Bone erosions are an important pathological feature of rheumatoid arthritis. The purpose of this work is to introduce a training tool to provide users with guidance on identifying pathological cortical breaks on high resolution peripheral quantitative computed tomography images for erosion analysis. Biology Development of a Rabbit Chronic-Like Rotator Cuff Injury Model for Study of Fibrosis and Muscular Fatty Degeneration Ke Li1,2,3, Xu Zhang1,2, Dan Wang1,2,3,4,5, Dai Fei Elmer Ker1,2,3,4,5 1School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, 2Institute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong Kong, 3Center for Neuromusculoskeletal Restorative Medicine, The Chinese University of Hong Kong, 4Key Laboratory for Regenerative Medicine, Ministry of Education, School of Biomedical Sciences, The Chinese University of Hong Kong, 5Department of Orthopaedics and Traumatology, Faculty of Medicine, The Chinese University of Hong Kong This study details procedures for establishing a chronic-like rabbit rotator cuff (RC) injury. Specifically, the injury is created in the subscapularis (SSC) muscle-tendon/myotendinous unit to mimic human RC anatomy and pathophysiology, including severe muscle fatty degeneration (FD). This protocol can be applied to study RC injuries and assess regenerative therapies. Bioengineering Prospective, Randomized, and Controlled Study of a Human Umbilical Cord Mesenchymal Stem Cell Injection for Treating Diabetic Foot Ulcers Jingyu Zhang*1,2, Bingkun Zhao*1, Wuhan Wei*1,2, Dan Wang3,4,5, Haoyu Wang1,2, Aijun Zhang1, Changbo Tao1, Xueyang Li1, Qiang Li1, Peisheng Jin1 1Department of Plastic Surgery, Affiliated Hospital of Xuzhou Medical University, 2Graduate School, Xuzhou Medical University, 3Institute for Tissue Engineering and Regenerative Medicine, The Chinese University of Hong Kong, 4School of Biomedical Sciences, The Chinese University of Hong Kong, 5Center for Neuromusculoskeletal Restorative Medicine, Hong Kong Science Park The present protocol describes a prospective, randomized, controlled clinical study that evaluates a human umbilical cord mesenchymal stem cell injection for the treatment of chronic diabetic foot ulcers. Bioengineering Creation of a Knee Joint-on-a-Chip for Modeling Joint Diseases and Testing Drugs Meagan J. Makarcyzk1,2, Zhong Alan Li1,3, Ilhan Yu1, Haruyo Yagi1, Xiurui Zhang1, Lauren Yocum1, Eileen Li1, Madalyn R. Fritch1, Qi Gao4, Bruce A. Bunnell5, Stuart B. Goodman4,6, Rocky S. Tuan1,8, Peter G. Alexander1,7, Hang Lin1,2,7 1Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, 2Department of Bioengineering, University of Pittsburgh Swanson School of Engineering, 3Department of Neurobiology, University of Pittsburgh School of Medicine, 4Department of Orthopaedic Surgery, Stanford University, 5Department of Microbiology, Immunology, and Genetics, University of North Texas Health Science Center, 6Department of Bioengineering, Stanford University, 7McGowan Institute for Regenerative Medicine, University of Pittsburgh School of Medicine, 8The Chinese University of Hong Kong We provide detailed methods for generating four types of tissues from human mesenchymal stem cells, which are used to recapitulate the cartilage, bone, fat pad, and synovium in the human knee joint. These four tissues are integrated into a customized bioreactor and connected through microfluidics, thus generating a knee joint-on-a-chip. Cancer Research Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma Kel Vin Tan*1,2, Xinxiang Yang*3, Chung Ying Chan2, Jingjing Shi1, Hing-Chiu Chang4, Keith Wan-Hang Chiu1,5, Kwan Man3 1Department of Diagnostic Radiology, School of Clinical Medicine, LKS Faculty of Medicine, The University of Hong Kong, 2Department of Oncology, MRC Oxford Institute for Radiation Oncology, University of Oxford, 3Department of Surgery, School of Clinical Medicine, HKU-SZH & LKS Faculty of Medicine, The University of Hong Kong, 4Department of Biomedical Engineering, The Chinese University of Hong Kong, 5Department of Diagnostic and Interventional Radiology, Kwong Wah Hospital Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG). Medicine Application of Optical Coherence Tomography to a Mouse Model of Retinopathy Xiaoting Mai*1, Shaofen Huang*1, Wen Chen1, Tsz Kin Ng1,2,3, Haoyu Chen1 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. Biology Multiplexed Fluorescent Immunohistochemical Staining of Four Endometrial Immune Cell Types in Recurrent Miscarriage Yiwei Zhao1, Gene Chi Wai Man1,2, Loucia Kit Ying Chan1, Xi Guo1, Yingyu Liu3, Tao Zhang1, Joseph Kwong1,4, Chi Chiu Wang1,5, Xiaoyan Chen3, Tin Chiu Li1 1Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Prince of Wales Hospital, 2Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong, Prince of Wales Hospital, 3Department of Obstetrics and Gynaecology, Shenzhen Baoan Women’s and Children’s Hospital, Shenzhen University, 4School of Medicine, Faculty of Medicine and Health Sciences, Keele University, 5Li Ka Shing Institute of Health Sciences; School of Biomedical Sciences, The Chinese University of Hong Kong Despite the advancements in multiplex immunohistochemistry and multispectral imaging, characterizing the density and clustering of major immune cells simultaneously in the endometrium remains a challenge. This paper describes a detailed multiplex staining protocol and imaging for the simultaneous localization of four immune cell types in the endometrium. Biology Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [18F]FDG Micro-PET/MR Imaging Qing Liu*1,2, Kel Vin Tan*3, Hing-Chiu Chang3, Pek-Lan Khong3, Xiaoyan Hui1,2,4 1State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, 2Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 3Department of Diagnostic Radiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 4School of Biomedical Sciences, Institute of Vascular Medicine, Chinese University of Hong Kong Functional imaging and quantitation of thermogenic adipose depots in mice using a micro-PET/MR imaging-based approach. Biochemistry Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells Wai Wa Ray Chan*1, Yu Qi Zhai*1, Kwok-Fai Lau1 1School of Life Sciences, The Chinese University of Hong Kong This paper aims to present a protocol for preparing recycling endosomes from mammalian cells using sucrose density gradient ultracentrifugation. Behavior Evaluation of the Cognitive Performance of Hypertensive Patients with Silent Cerebrovascular Lesions Manman Zhang1,2, Junling Gao3, Bingjiao Xie2, Henry Ka Fung Mak4, Raymond Tak Fai Cheung2,5 1Department of Neurology, The First Affiliated Hospital of Wenzhou Medical University, 2Department of Medicine, The University of Hong Kong, 3Centre of Buddhist Studies, The University of Hong Kong, 4Department of Diagnostic Radiology, The University of Hong Kong, 5Research Centre of Heart, Brain, Hormone & Healthy Aging, The University of Hong Kong Here, we present a protocol to assess whether various types of silent cerebrovascular lesions are differentially associated with deficits in certain cognitive domains in a cohort of 398 hypertensive elderly Chinese, using a combination of neuropsychological tests and multi-sequence 3T MRI scanning. Developmental Biology An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons Wai Wa Ray Chan*1, Wen Li*1,2, Dik Long Dennis Chau*1, Kwok-Fai Lau1 1School of Life Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, 2Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen In this report, we describe a simple protocol for studying neurite outgrowth in embryonic rat cortical neurons by co-transfecting with EGFP and the protein of interest. Neuroscience Characterizing the Relationship Between Eye Movement Parameters and Cognitive Functions in Non-demented Parkinson's Disease Patients with Eye Tracking Oscar W.H. Wong1, Gabriel P.C. Fung2, Sandra Chan1 1Department of Psychiatry, The Chinese University of Hong Kong, 2Lab Viso Limited Here, we present a protocol to study the relationship between the eye movement parameters and cognitive functions in non-demented Parkinson's disease patients. The experiment used an eye tracker to measure the saccadic amplitude and fixation duration in a visual search task. The correlation with performance in multi-domain cognitive tasks was subsequently measured. Genetics A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene Huilin Wang*1, Xiaofan Zhu*2,3, Baoheng Gui*3,4, Wan Chee Cheung2, Mengmeng Shi2,3, Zhenjun Yang2,3, Ka Yin Kwok2, Ricky Lim5, Sanna Pietilä5, Yuanfang Zhu6,7, Kwong Wai Choy2,3 1Central Laboratory, Bao'an Maternity and Child Health Hospital, Jinan University, 2Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, 3Shenzhen Research Institute, The Chinese University of Hong Kong, 4Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, 5PerkinElmer Diagnostics, 6Department of Obstetrics and Gynecology, Bao'an Maternity and Child Health Hospital, Jinan University, 7Maternal-Fetal Medicine Institute, Bao'an Maternity and Child Health Hospital, Jinan University An accurate and robust polymerase chain reaction-based assay for quantifying cytosine-guanine-guanine trinucleotide repeats in the Fragile X mental retardation-1 gene facilitates molecular diagnosis and screening of Fragile X syndrome and Fragile X-related disorders with shorter turn-around time and investment in equipment. Genetics Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells Chi-Keung Cheng1, Terry H.Y. Wong1, Yuk-Lin Yung1, Nelson C.N. Chan1, Margaret H.L. Ng1,2 1Blood Cancer Cytogenetics and Genomics Laboratory, Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, 2State Key Laboratory in Oncology in South China, The Chinese University of Hong Kong Direct delivery of preassembled Cas9/guide RNA ribonucleoprotein complexes is a fast and efficient means for genome editing in hematopoietic cells. Here, we utilize this approach to delete a RUNX1 intronic silencer and examine the transcriptional responses in OCI-AML3 leukemic cells. Genetics Semiconductor Sequencing for Preimplantation Genetic Testing for Aneuploidy Baoheng Gui*1,2,3, Yingxin Zhang*4, Bo Liang5, Yvonne Ka Yin Kwok4, Wai Ting Lui4, Queenie Sum Yee Yeung4, Lingyin Kong6, Liming Xuan6, Jacqueline Pui Wah Chung4, Kwong Wai Choy3,4 1Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, 2Birth Defects Prevention and Control Institute of Guangxi Zhuang Autonomous Region, 3Shenzhen Research Institute, The Chinese University of Hong Kong, 4Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, 5State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 6Basecare Medical Device Co., Ltd Here, we introduce a semiconductor sequencing method for preimplantation genetic testing for aneuploidy (PGT-A) with the advantages of short turnaround time, low cost, and high throughput. Cancer Research Flow Cytometric Detection of Newly-formed Breast Cancer Stem Cell-like Cells After Apoptosis Reversal Yiyue Xu1,2, Chun So1, Hon-Ming Lam1,2, Ming-Chiu Fung1,2, Suk-Ying Tsang1,2,3,4 1School of Life Sciences, The Chinese University of Hong Kong, 2State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, 3Key Laboratory for Regenerative Medicine, Ministry of Education, The Chinese University of Hong Kong, 4Centre for Novel Biomaterials, The Chinese University of Hong Kong Here, we present a protocol to isolate apoptotic breast cancer cells by fluorescence-activated cell sorting and further detect the transition of breast non-stem cancer cells to breast cancer stem cell-like cells after apoptosis reversal by flow cytometry. Developmental Biology A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes Jiayin Yang*1,2,6, Lai-Yung Wong*2, Xiao-Yu Tian*3, Rui Wei1,2, Wing-Hon Lai2, Ka-Wing Au2, Zhiwei Luo4,5, Carl Ward4,5, Wai-In Ho2, David P. Ibañez4,5, Hao Liu4,5, Xichen Bao4,5, Baoming Qin4,5, Yu Huang3, Miguel A. Esteban4,5,7, Hung-Fat Tse1,2,6,7 1Department of Medicine, University of Hong Kong-Shenzhen Hospital, 2The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, 3School of Biomedical Sciences, Institute of Vascular Medicine, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, 4Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical University, 5Laboratory of RNA, Chromatin, and Human Disease, CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 6Research Centre of Heart, Brain, Hormone, and Healthy Ageing, Li Ka Shing Faculty of Medicine, University of Hong Kong, 7Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, University of Hong Kong and Guangzhou Institutes of Biomedicine and Health Here, we present a protocol to generate a human liver chimeric mouse model of familial hypercholesterolemia using human induced pluripotent stem cell-derived hepatocytes. This is a valuable model for testing new therapies for hypercholesterolemia. Biology Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model Zhaofeng Jia1,2,3, Yujie Liang4,5, Xingfu Li2,3, Xiao Xu1,2,3, Jianyi Xiong2,3, Daping Wang1,2,3, Li Duan2,3 1Postgraduate institution, Guangzhou Medical University, 2Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, 3 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. Engineering Determination of the Excitation and Coupling Rates Between Light Emitters and Surface Plasmon Polaritons Zhaolong Cao1,2, Min Lin2, Daniel Ong2 1State Key Laboratory of Optoelectronic Materials and Technologies and Guangdong Province Key Laboratory of Display Material and Technology, School of Physics and Engineering, Sun Yat-sen University, 2Department of Physics, The Chinese University of Hong Kong This protocol describes the instrumentation for determining the excitation and coupling rates between light emitters and Bloch-like surface plasmon polaritons arising from periodic arrays. Medicine Detecting Anastasis In Vivo by CaspaseTracker Biosensor Ho Man Tang1,2, Ming Chiu Fung2, Ho Lam Tang3 1Institute for Basic Biomedical Sciences, Johns Hopkins University School of Medicine, 2School of Life Sciences, Chinese University of Hong Kong, 3Department of Neurosurgery, Johns Hopkins University School of Medicine Anastasis is technically challenging to detect in vivo because the cells that have reversed the cell death process can be morphologically indistinguishable from normal healthy cells. Here we describe protocols for detecting and tracking cells that undergo anastasis in live animals by using our newly developed in vivo CaspaseTracker biosensor system. Immunology and Infection A Novel Feeder-free System for Mass Production of Murine Natural Killer Cells In Vitro Patrick Ming-Kuen Tang1,2, Philip Chiu-Tsun Tang2, Jeff Yat-Fai Chung2, Jessica Shuk Chun Hung2, Qing-Ming Wang2, Guang-Yu Lian2, Jingyi Sheng2, Xiao-Ru Huang2, Ka-Fai To1, Hui-Yao Lan2 1Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, 2Li Ka Shing Institute of Health Sciences and Department of Medicine & Therapeutics, The Chinese University of Hong Kong Here, we present a protocol to mass-produce gene-silencing murine NK cells by using a feeder-free differentiation system for mechanistic study in vitro and in vivo. Medicine Using Retinal Imaging to Study Dementia Victor T.T. Chan1, Tiffany H.K. Tso1, Fangyao Tang1, Clement Tham1, Vincent Mok2,3,4, Christopher Chen5,6, Tien Y. Wong7,8, Carol Y. Cheung1 1Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, 2Department of Medicine & Therapeutics, The Chinese University of Hong Kong, 3Therese Pei Fong Chow Research Centre for Prevention of Dementia, The Chinese University of Hong Kong, 4Gerald Choa Neuroscience Centre, The Chinese University of Hong Kong, 5Memory Aging and Cognition Centre, National University Health System, 6Department of Pharmacology, National University of Singapore, 7Singapore Eye Research Institute, Singapore National Eye Centre, 8Duke-NUS Medical School, National University of Singapore The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia. Developmental Biology A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro Zhaofeng Jia1,2, Yujie Liang3, Bin Ma4,5, Xiao Xu2,6, Jianyi Xiong2, Li Duan2, Daping Wang1,2 1Guangzhou Medical University, 2 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. Biology In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila Ho Lam Tang1, Ho Man Tang1, Ming Chiu Fung2, J. Marie Hardwick1 1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 2School of Life Sciences, Chinese University of Hong Kong To detect healthy cells in whole animals that contain low levels of caspase activity, the highly sensitive biosensor designated CaspaseTracker was generated for Drosophila. Caspase-dependent biosensor activity is detected in long-lived healthy cells throughout the internal organs of adult animals reared under optimized conditions in the absence of death stimuli. Biology Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis Ho Lam Tang1,2,3, Ho Man Tang1,2,3, J. Marie Hardwick1, Ming Chiu Fung2 1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 2School of Life Sciences, Chinese University of Hong Kong, 3Center for Cell Dynamics, Department of Biological Chemistry, Johns Hopkins University School of Medicine The term anastasis refers to the phenomenon in which dying cells reverse a cell suicide process at a late stage, repair themselves, and ultimately survive. Here we demonstrate protocols for detecting and tracking cells that undergo anastasis.
