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Soochow University

21 ARTICLES PUBLISHED IN JoVE

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Biology

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure
Kenneth K. Kwong 1,2, Lenuta Kloetzer 1,2,3,4, Kelvin K. Wong 5,6, Jia-Qian Ren 1,2, Braden Kuo 1,2,3,4, Yan Jiang 7, Y. Iris Chen 1,2, Suk-Tak Chan 1,2,8, Geoffrey S. Young 9, Stephen T.C. Wong 5,6
1Department of Radiology, Massachusetts General Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 3Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, 4Department of Medicine, Massachusetts General Hospital, Harvard Medical School, 5Center for biotechnology and Informatics, The Methodist Hospital Research Institute, 6Department of Radiology, The Methodist Hospital, Weill Cornell Medical College, 7Bejing University of Chinese Medicine, 8Department of Health Technology and Informatics, The Hong Kong Polytechnic University, 9Department of Radiology, Brigham and Women's Hospital, Harvard Medical School

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

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Biology

In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes
Yanning Wu 1, Shuo Wang 1, Chunying Li 1,2,3
1Department of Biochemistry & Molecular Biology, Wayne State University School of Medicine, 2Cardiovascular Research Institute, Wayne State University School of Medicine, 3Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine

Cystic fibrosis transmembrane conductance regulator (CFTR), an epithelial chloride channel, has been reported to interact with various proteins and regulate important cellular processes; among them the CFTR PDZ motif-mediated interactions have been well documented. This protocol describes methods we developed to assemble a PDZ-dependent CFTR macromolecular signaling complex in vitro.

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Behavior

Creating Dynamic Images of Short-lived Dopamine Fluctuations with lp-ntPET: Dopamine Movies of Cigarette Smoking
Evan D. Morris 1,2,3,4, Su Jin Kim 1,3, Jenna M. Sullivan 1,3,4, Shuo Wang 3,4, Marc D. Normandin 5, Cristian C. Constantinescu 6, Kelly P. Cosgrove 1,2,3
1Diagnostic Radiology, Yale University, 2Psychiatry, Yale University, 3Yale PET Center, Yale University, 4Biomedical Engineering, Yale University, 5Nuclear Medicine, Massachusetts General Hospital, 6Radiological Sciences, University of California, Irvine

We present a novel PET imaging approach for capturing dopamine fluctuations induced by cigarette smoking. Subjects smoke in the PET scanner. Dynamic PET images are modeled voxel-by-voxel in time by lp-ntPET, which includes a time-varying dopamine term. The results are 'movies' of dopamine fluctuations in the striatum during smoking.

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Developmental Biology

Isolation and Cryopreservation of Neonatal Rat Cardiomyocytes
Adam C. Vandergriff 1,2, Michael Taylor Hensley 1,2, Ke Cheng 1,2,3
1Department of Molecular Biomedical Sciences and Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, 2Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill, North Carolina State University, 3The Cyrus Tang Hematology Center, Soochow University

The isolation of neonatal rat cardiomyocytes is a time consuming and unpredictable procedure. This study describes methods for cryopreservation and thawing of neonatal rat cardiomyocytes that allows for more efficient use of cells. The thawed NRCMs can be used for various experiments without the need for performing isolations each time.

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Biochemistry

Regeneration of Arrayed Gold Microelectrodes Equipped for a Real-Time Cell Analyzer
Zhihui Xu *1, Yiyan Song *1, Huijun Jiang 2, Yan Kong 3, Xiaoming Li 3, Jin Chen 1, Yuan Wu 4
1School of Public Health, Nanjing Medical University, 2School of Pharmacy, Nanjing Medical University, 3State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing Tech University, 4Department of Medical Oncology, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University

This protocol describes a general strategy to regenerate commercial arrayed gold microelectrodes equipped for a label-free cell analyzer aimed at saving on the high running costs ofmicrochip-based assays. The regeneration process includes trypsin digestion, rinsing with ethanol and water, and a spinning step, which enables repeated usage of microchips.

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Biology

In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability
Ban-Ruo Li *1,2, Jia Wu *1,2, Hua-Shan Li 1,2, Zhi-Hui Jiang 1,2, Xiu-Min Zhou 1,2, Cai-Hua Xu 1,2, Ning Ding 1,2, Juan-Min Zha 1,2, Wei-Qi He 1,2
1Jiangsu Key Laboratory of Neuropsychiatric Diseases and Cambridge-Suda (CAM-SU) Genome Resource Center, Soochow University, 2Department of Oncology, The First Affiliated Hospital of Soochow University

Two methods are presented here to determine intestinal barrier function. An epithelial meter (volt/ohm) is used for measurements of transepithelial electrical resistance of cultured epithelia directly in tissue culture wells. In mice, the FITC-dextran gavage method is used to determine the intestinal permeability in vivo.

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Behavior

Simultaneous Eye Tracking and Single-Neuron Recordings in Human Epilepsy Patients
Shuo Wang 1, Nand Chandravadia 2, Adam N. Mamelak 2, Ueli Rutishauser 2,3,4
1Department of Chemical and Biomedical Engineering, and Rockefeller Neuroscience Institute, West Virginia University, 2Departments of Neurosurgery and Neurology, Cedars-Sinai Medical Center, 3Center for Neural Science and Medicine, Department of Biomedical Sciences, Cedars-Sinai Medical Center, 4Division of Biology and Biological Engineering, California Institute of Technology

We describe a method to conduct single-neuron recordings with simultaneous eye tracking in humans. We demonstrate the utility of this method and illustrate how we used this approach to obtain neurons in the human medial temporal lobe that encode targets of a visual search.

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Behavior

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze
Huiling Tian 1, Ning Ding 2, Mengwei Guo 1, Shun Wang 1, Zidong Wang 1, Hao Liu 1, Jiayi Yang 1, Yujie Li 3, Jingyu Ren 1, Jing Jiang 4, Zhigang Li 1
1School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, 2Guang'anmen Hospital of China Academy of Chinese Medical Science, 3The Third Affiliated Hospital of Beijng University of Chinese Medicine, 4School of Nursing, Beijing University of Chinese Medicine

Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer’s Disease model mice and to assess the effect of manual acupuncture for treating them is described.

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Biology

Quantification of Proliferative and Dead Cells in Enteroids
Hua-Shan Li *1, Shao-Fang Xu *1, Jian-Ying Sheng *1, Zhi-Hui Jiang 1, Jing Wang 1, Ning Ding 1, Tao Wang 1, Matthew A. Odenwald 2, Jerrold R. Turner 2,3, Wei-Qi He 1, Hong Xu 1, Juan-Min Zha 1
1Jiangsu Key Laboratory of Neuropsychiatric Diseases and Cambridge-Suda (CAM-SU) Genomic Resource Center, Medical College of Soochow University, Department of Oncology, The First Affiliated Hospital of Soochow University, 2Department of Pathology, University of Chicago, 3Department of Pathology, Brigham and Women's Hospital–Harvard Medical School

The presented protocol uses flow cytometry to quantify the number of proliferating and dead cells in cultured mouse enteroids. This method is helpful to evaluate the effects of drug treatment on organoid proliferation and survival.

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Cancer Research

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging
Nour Ghaddar 1,2, Shuo Wang 1, Véronique Michaud 1, Urszula Kazimierczak 1,3, Nicolas Ah-son 1, Antonis E. Koromilas 1,4
1Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, 2Division of Experimental Medicine, Faculty of Medicine, McGill University, 3Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, 4Gerald Bronfman Department of Oncology, Faculty of Medicine, McGill University

This protocol describes the steps taken to induce KRAS lung tumors in mice as well as the quantification of formed tumors by ultrasound imaging. Small tumors are visualized in early timepoints as B-lines. At later timepoints, relative tumor volume measurements are achieved by the measurement tool in the ultrasound software.

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Biology

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy
Ayse Sena Mutlu 1, Tao Chen 1, Dinghuan Deng 1, Meng C. Wang 1,2,3
1Huffington Center on Aging, Baylor College of Medicine, 2Department of Molecular and Human Genetics, Baylor College of Medicine, 3Howard Hughes Medical Institute, Baylor College of Medicine

Stimulated Raman scattering (SRS) microscopy allows selective, label-free imaging of specific chemical moieties and it has been effectively employed to image lipid molecules in vivo. Here, we provide a brief introduction to the principle of SRS microscopy and describe methods for its use in imaging lipid storage in Caenorhabditis elegans.

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Developmental Biology

Using Immunofluorescence to Detect PM2.5-induced DNA Damage in Zebrafish Embryo Hearts
Yujie Huang *1, Yizhou Tao *1, Chang Cai 1, Jin Chen 1, Cheng Ji 1, Stanley Aniagu 2, Yan Jiang 1, Tao Chen 1
1School of Public Health, Soochow University, 2Toxicology, Risk Assessment and Research Division, Texas Commission on Environmental Quality

This protocol uses an immunofluorescence assay to detect PM2.5-induced DNA damage in the dissected hearts of zebrafish embryos.

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Neuroscience

Isolation of Adult Human Astrocyte Populations from Fresh-Frozen Cortex Using Fluorescence-Activated Nuclei Sorting
Zarmeen Mussa 1,2, Jessica Tome-Garcia 1,2, Yan Jiang 3, Schahram Akbarian 2,4, Nadejda M. Tsankova 1,2
1Department of Pathology, Icahn School of Medicine at Mount Sinai, 2Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, 3Institutes of Brian Science, Fudan University, 4Department of Psychiatry, Icahn School of Medicine at Mount Sinai

We have developed a method that enriches for and isolates human astrocyte populations from fresh-frozen tissue for use in downstream molecular analyses.

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Medicine

Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct
Xiaochong Zhou 1, Haifeng Chen 1, Xing Wei 1, Yang He 2,4,5, Chunfang Xu 1, Zhen Weng 2,3,4,6
1Department of Gastroenterology, the First Affiliated Hospital of Soochow University, 2MOE Engineering Center of Hematological Disease, Soochow University, 3Cyrus Tang Hematology Center, Soochow University, 4National Clinical Research Center for Hematologic Diseases, the First Affiliated Hospital of Soochow University, 5MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, 6Collaborative Innovation Center of Hematology, Soochow University

A mouse model of severe acute pancreatitis is described herein. The procedure presented here is very rapid, simple, and accessible, thereby potentially allowing the study of the molecular mechanisms and different therapeutic interventions in acute pancreatitis in a convenient way.

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Medicine

Isolation of Monocyte-Macrophage Lineage Cells from Rat Bones by Secondary Adherence Method
Xiaoli Jin *1, Yang Li *2, Xuanwei Chen 1, Jin Chen 1, Jian Xu 1
1School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, 2School of Basic Medical Sciences, Fudan University

Here we present a protocol for the isolation of BMMs from SD rats, called the secondary adherence method.

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Developmental Biology

Isolation and Culture of Three Kinds of Umbilical Cord Mesenchymal Stem Cells
Yanjun Yang *1, Chunxue Zhang *2, Xiaoqiang Sheng 3
1Department of Obstetrics and Gynecology, the Third Affiliated Hospital, Soochow University, 2Department of Obstetrics and Gynecology, Nanjing First Hospital, Nanjing Medical University, 3Center for Reproductive Medicine and Obstetrics and Gynecology, Nanjing Drum Tower Hospital, Nanjing University Medical School

The present protocol describes the isolation and culture of mesenchymal stem cells from the umbilical cord arteries, vein, and Wharton's jelly.

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Developmental Biology

Mitochondrial Transfer from Mouse Adipose-Derived Mesenchymal Stem Cells into Aged Mouse Oocytes
Yanjun Yang 1, Chunxue Zhang 2, Xiaoqiang Sheng 3
1Department of Obstetrics and Gynecology, the Third Affiliated Hospital, Soochow University, 2Department of Obstetrics and Gynecology, Nanjing First Hospital, Nanjing Medical University, 3Center for Reproductive Medicine and Obstetrics and Gynecology, Nanjing Drum Tower Hospital, Nanjing University Medical School

Here, we describe the isolation of mitochondria from mouse adipose-derived mesenchymal stem cells, and then transfer the mitochondria into aged mouse oocytes to improve the quality of the oocytes.

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Biology

A Mouse Model of Ankle-Subtalar Complex Joint Instability
Shuo Wang *1,2, Peixin Liu *1,2, Chunzhuo Hua 1,2, Hongtao Zhang 1,2, Jia Yu 1,2
1Department of Orthopaedics, The First Affiliated Hospital of Soochow University, 2Orthopaedic Institute, Soochow University

The ankle-subtalar complex joint (ASCJ) is the core of the foot and plays a key role in balance control in daily activities. Sports injuries often lead to instability in this joint. Here, we describe a mouse model of ligament transection-induced instability of the ASCJ.

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Current Techniques and Models for Assessing the Developmental Toxicity of Environmental Pollutants
Yan Jiang 1,2, Stanley Aniagu 3, Tao Chen 1,2
1Suzhou Medical College, Soochow University, 2Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow University, 3Toxicology, Risk Assessment, and Research Division, Texas Commission on Environmental Quality

Current Techniques and Models for Assessing the Developmental Toxicity of Environmental Pollutants

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Behavior

A Simple Technique to Assay Locomotor Activity in Drosophila
Xiang Long 1, Huanhuan Du 1, Mengni Jiang 1, Hongrui Meng 1,2
1Institute of Neuroscience, Soochow University, 2Department of Neurology and Clinical Research Center of Neurological Disease, The Second Affiliated Hospital of Soochow University

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

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Biology

Preparation of a Single-Cell Suspension from Mouse Carotid Arteries for Single-Cell Sequencing
Fengchan Li 1, Zhen Zhu 1, Yun Du 1, Li Zhu 1,2,3, Chaojun Tang 1,2,3
1Cyrus Tang Hematology Center, Cyrus Tang Medical Institute, Soochow University, 2Collaborative Innovation Center of Hematology of Jiangsu Province, Soochow University, 3Suzhou Key Laboratory of Thrombosis and Vascular Biology, Soochow University

Here we describe a two-step cell digestion protocol for preparing a single-cell suspension of mouse carotid arteries.

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