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Qilu hospital of shandong university

14 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)
Ronald W. Jensen 1, Jason Rivest 1, Wei Li 1, Varalakshmi Vissa 1
1Department of Microbiology, Immunology and Pathology, Colorado State University

Leprosy, caused by Mycobacterium leprae, is still endemic in many places. In order to learn about the spread and mode of transmission of leprosy, it is important to determine which strain of M. leprae has infected a patient. Variable numbers of tandem repeats (VNTR) typing is one such method.

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Medicine

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies
Wei Li 1, Shu Zhu 1, Yusong Zhang 1, Jianhua Li 1, Andrew E. Sama 1, Ping Wang 1, Haichao Wang 1
1The Feinstein Institute for Medical Research, North Shore – LIJ Health System

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

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JoVE Core

SSVEP-based Experimental Procedure for Brain-Robot Interaction with Humanoid Robots
Jing Zhao 1, Wei Li 1,2, Xiaoqian Mao 1, Mengfan Li 1
1School of Electrical Engineering and Automation, Tianjin University, 2Department of Computer & Electrical Engineering and Computer Science, California State University

The overall goal of this method is to establish an SSVEP-based experimental procedure by integrating multiple software programs to enable the study of brain-robot interaction with humanoid robots, which is prospective in assisting the sick and elderly as well as performing unsanitary or dangerous jobs.

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Medicine

Ferric Chloride-induced Murine Thrombosis Models
Wei Li 1,2, Marvin Nieman 3, Anirban Sen Gupta 4
1Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, 2Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, 3Department of Pharmacology, Case Western Reserve University, 4Department of Biomedical Engineering, Case Western Reserve University

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

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Neuroscience

Mitochondrial Ca2+ Retention Capacity Assay and Ca2+-triggered Mitochondrial Swelling Assay
Wei Li 1,2, Chen Zhang 1, Xiulian Sun 3
1Department of Neurology, Qilu Hospital of Shandong University, 2Department of Neurology, Qingdao Municipal Hospital, 3Brain Research Institute, Qilu Hospital of Shandong University

This protocol aims to describe a method to examine the Ca2+ retention capacity and Ca2+- triggered mitochondrial swelling of isolated mitochondria of SH-SY5Y cells step-by-step.

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Developmental Biology

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue
Zhenan Liu 1, Jie Wen 1,2, Xue Leng 1, Qian Zhou 1, Changkuo Zhou 3, Huaqiang Zhao 1, Xunwei Wu 1,2
1Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Laboratory for Tissue Engineering and Regeneration, School of Stomatology, Shandong University, 2Suzhou Institute of Shandong University, 3Department of Urology, Qilu Hospital of Shandong University

Here we present a protocol to efficiently isolate primary human keratinocytes from adult skin tissues. This method simplifies the conventional procedure by using the ROCK Inhibitor Y-27632 in the inoculation medium to spontaneously separate epidermal cells from dermal cells.

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Biochemistry

Preparation of Fungal and Plant Materials for Structural Elucidation Using Dynamic Nuclear Polarization Solid-State NMR
Alex Kirui *1, Malitha C. Dickwella Widanage *1, Frederic Mentink-Vigier 2, Ping Wang 3, Xue Kang 1, Tuo Wang 1
1Department of Chemistry, Louisiana State University, 2National High Magnetic Field Laboratory, 3Departments of Pediatrics, and Microbiology, Immunology and Parasitology, Louisiana State University Health Sciences Center

A protocol for preparing 13C,15N-labeled fungal and plant samples for multidimensional solid-state NMR spectroscopy and dynamic nuclear polarization (DNP) investigations is presented.

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Biochemistry

A Semi-Quantitative Drug Affinity Responsive Target Stability (DARTS) assay for studying Rapamycin/mTOR interaction
Chen Zhang *1, Min Cui *1, Yazhou Cui 1, Aubryanna Hettinghouse 1, Chuan-ju Liu 1,2
1Department of Orthopaedic Surgery, New York University Medical Center, 2Department of Cell Biology, New York University School of Medicine

In this study, we enhanced the data analysis capabilities of the DARTS experiment by monitoring the changes in protein stability and estimating the affinity of protein-ligand interactions. The interactions can be plotted into two curves: a proteolytic curve and a dose-dependence curve. We have used mTOR-rapamycin interaction as an exemplary case.

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Biology

Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues
Chang Liu 1, Shuangshuang Wang 1, Man Liu 2, Fuxiang Bai 1, Zhihong Chen 3, Ping Wang 4, Jun Mi 1,5, Xunwei Wu 1
1Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 2Shijiazhuang Shimen Experimental School, 3Qilu Children's Hospital of Shandong University, 4Qilu Hospital of Shandong University, 5Shenzhen Research Institute of Shandong University

This paper reports that the addition of Y-27632 to TIVA medium can significantly increase the yield of melanocytes from adult skin tissues.

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Chemistry

Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium
Kiyoharu J. Miyagishima 1, Congxiao Zhang *2, Volha V. Malechka *3, Kapil Bharti 2, Wei Li 1
1Retinal Neurophysiology Section, National Eye Institute, National Institutes of Health, 2Ocular and Stem Cell Translational Research Section, National Eye Institute, National Institutes of Health, 3Human Visual Function Core, National Eye Institute, National Institutes of Health

Here, we present a method for recording light-evoked electrical responses of the retinal pigment epithelium (RPE) in mice using a technique known as DC-ERGs first described by Marmorstein, Peachey, and colleagues in the early 2000s.

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Engineering

Design and Development of a Three-Dimensionally Printed Microscope Mask Alignment Adapter for the Fabrication of Multilayer Microfluidic Devices
Celine R. Garcia *1, Zhenya Ding *1, Hilario C. Garza 1, Wei Li 1
1Department of Chemical Engineering, Texas Tech University

This project allows small laboratories to develop an easy-to-use platform for the fabrication of precise multilayer microfluidic devices. The platform consists of a three-dimensionally printed microscope mask alignment adapter using which multilayer microfluidic devices with alignment errors of <10 µm were achieved.

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Biology

Primary Culture of Porcine Retinal Pigment Epithelial Cells
Feng Wen *1,2,3, Yanzi Wang *1,2,3, Danxue He 1,2,3, Chunyan Liao 1,2,3, Weijie Ouyang 1,2,3, Zuguo Liu 1,2,3, Wei Li 1,2,3, Yi Liao 1,2,3
1Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, 2Department of Ophthalmology, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, 3Xiamen University Affiliated Xiamen Eye Center, School of Medicine, Xiamen University

Here, an easy-to-follow method to culture primary porcine retinal pigment epithelial cells in vitro is presented.

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Neuroscience

Establishing a Mouse Contusion Spinal Cord Injury Model Based on a Minimally Invasive Technique
Elham Yilizati-Yilihamu Elzat 1, Xiangchuang Fan 1, Zimeng Yang 1, Zhongze Yuan 1, Yilin Pang 2, Shiqing Feng 1,2
1Department of Orthopaedics, Qilu Hospital of Shandong University, 2Department of Orthopaedics, Tianjin Medical University General Hospital

Minimally invasive techniques and a simple laboratory device improve the reproducibility of the spinal cord injury model by reducing operative damage to the experimental animals and allowing anatomical morphology maintenance. The method is worthwhile because the reliable results and reproducible procedure facilitate investigations of the mechanisms of disease reparation.

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Neuroscience

Establishment of Central Cord Syndrome Model in C57BL/6J Mouse
Elham Yilizati-Yilihamu Elzat *1, Xiangchuang Fan *1, Shiqing Feng 1,2
1Department of Orthopaedics, Qilu Hospital of Shandong University, 2Department of Orthopaedics, Tianjin Medical University General Hospital

The present protocol simulating central cord syndrome (CCS) in mice has improved repeatability and minimized operation damage to the experimental animals, avoiding disrupting the anatomical structure excessively. The strategy in this study is advantageous because it allows for research into injury mechanisms by producing consistent results.

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