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The First Affiliated Hospital of Soochow University

20 ARTICLES PUBLISHED IN JoVE

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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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Medicine

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots
Lu Xu 1, Qiwen Shen 1, Zhongqi Mao 2, L. James Lee 3, Ouliana Ziouzenkova 1
1Department of Human Sciences, The Ohio State University, 2Department of Minimally Invasive Surgery, The First Affiliated Hospital of Soochow University, 3NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

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Genetics

In Situ Labeling of Mitochondrial DNA Replication in Drosophila Adult Ovaries by EdU Staining
Zhe Chen 1, Hong Xu 1
1Lab of Molecular Genetics, National Heart, Lung and Blood Institute, National Institutes of Health

Drosophila oogenesis continues to be exceptionally useful in the study of mitochondrial proliferation and inheritance. This manuscript describes a detailed protocol used to label the replicating mitochondrial DNA (mtDNA) in Drosophila adult ovaries with 5-ethynyl-2´-deoxyuridine (EdU), which facilitates uncovering mechanisms associated with mitochondrial inheritance that were previously debatable.

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Genetics

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Muhammad Khairul Ramlee 1, Jing Wang 1, Alice M. S. Cheung 1, Shang Li 1
1Cancer & Stem Cell Biology Programme, Duke-NUS Medical School

The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.

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Neuroscience

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method
Thomas R. Groves 1,2,3, Jing Wang 1,2, Marjan Boerma 1,2, Antiño R. Allen 1,2,3
1Division of Radiation Health, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 3Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

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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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Neuroscience

Intracranial Pharmacotherapy and Pain Assays in Rodents
Erik Martinez 1, Haocheng Zhou 1, Jing Wang 1,2
1Department of Anesthesiology, Perioperative Care and Pain Medicine, New York University School of Medicine, 2Department of Neuroscience and Physiology, New York University School of Medicine

Here we present a protocol to perform intracranial pharmacological experiments followed by pain behavior assays in rodents. This protocol allows researchers to deliver molecular and cellular targets in the brain, for pharmacologic agents in the treatment of pain.

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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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Neuroscience

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation
Ming Luo 1, Xiangyue Tang 1, Juehua Zhu 2, Zhihua Qiu 1, Yongjun Jiang 1
1The Second Affiliated Hospital of Guangzhou Medical University, 2Department of Neurology, The First Affiliated Hospital of Soochow University

Presented here is a protocol for establishing acute pontine infarction in a rat model via electrical stimulation with a single pulse.

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Behavior

Implementation of a Real-Time Psychosis Risk Detection and Alerting System Based on Electronic Health Records using CogStack
Tao Wang 1, Dominic Oliver 2, Yamiko Msosa 1, Craig Colling 3, Giulia Spada 2, Łukasz Roguski 4, Amos Folarin 1, Robert Stewart 3,5, Angus Roberts 1,3, Richard J. B. Dobson 1,3,4,6, Paolo Fusar-Poli 2,3,7,8
1Department of Biostatistics and Health Informatics, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 2Early Psychosis: Interventions and Clinical-detection (EPIC) lab, Department of Psychosis Studies, Institute of Psychiatry, Psychology & Neuroscience, King's College London, 3National Institute for Health Research, Maudsley Biomedical Research Centre, South London and Maudsley National Health Service (NHS) Foundation Trust, 4Institute of Health Informatics, University College London, 5Department of Psychological Medicine, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 6Health Data Research UK London, University College London, 7OASIS service, South London and Maudsley National Health Service (NHS) Foundation Trust, 8Department of Brain and Behavioral Sciences, University of Pavia

We demonstrate how to deploy a real-time psychosis risk calculation and alerting system based on CogStack, an information retrieval and extraction platform for electronic health records.

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Medicine

Applying a Three-dimensional Uniaxial Mechanical Stimulation Bioreactor System to Induce Tenogenic Differentiation of Tendon-Derived Stem Cells
Ziming Chen *1, Peilin Chen *1, Rui Ruan *1, Lianzhi Chen 1, Jun Yuan 1, David Wood 1, Tao Wang 1, Ming Hao Zheng 1
1Centre of Orthopaedic Translational Research, Medical School, University of Western Australia

A three-dimensional uniaxial mechanical stimulation bioreactor system is an ideal bioreactor for tenogenic-specific differentiation of tendon-derived stem cells and neo-tendon formation.

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Medicine

Creating Radio-cephalic Arteriovenous Fistula in the Forearm with a Modified No-Touch Technique
Zhang Yuanyuan *1, Wang Xiaohe *1, Liu Zhen 1, Hou Guocun 1
1Department of Nephrology, The Affiliated Suzhou Science & Technology Town Hospital, Nanjing Medical University

We present a modified no-touch technique (MNTT) to create a radio-cephalic arteriovenous fistula (RC-AVF) in which the venous and arterial wall avoid devascularization and the radial artery does not sever.

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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

A Modified Technique for Arteriovenous Fistula Construction in Rabbits
Liu Zhen *1,2,3, Hou Guocun *3, Wang Xiaohe 3, Hu Jingfang 3, Li Jie 3, Wei Minggang 1,2
1Department of Traditional Chinese Medicine, The First Affiliated Hospital of Soochow University, 2Department of Nephrology, The First Affiliated Hospital of Soochow University, 3Department of Nephrology, Suzhou Science & Technology Town Hospital

The present protocol proposes the creation of an arteriovenous fistula in rabbits using a modified no-touch technique. The technique involves the side-to-side anastomosis of the common carotid artery and external jugular vein without the dissection of the perivenous tissues or cutting off the artery.

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Medicine

Establishing a Silicosis Rat Model via Exposure of Whole-Body to Respirable Silica
Fuyu Jin 1, Yaqian Li 1, Tian Li 1, Xinyu Yang 1, Wenchen Cai 1, Shifeng Li 1, Xuemin Gao 1, Fang Yang 1, Hong Xu 1, Heliang Liu 1
1Hebei Key Laboratory for Organ Fibrosis Research, School of Public Health, North China University of Science and Technology

This study describes a technique to establish a silicosis rat model with the inhalation of silica through the whole body in an inhalation chamber. The rats with silicosis could closely mimic the pathological process of human silicosis in an easy, cost-effective manner with good repeatability.

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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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