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49 ARTICLES PUBLISHED IN JoVE

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Biology

The use of SC1 (Pluripotin) to Support mESC Self-renewal in the Absence of LIF
Wen Xiong 1, Yan Gao 1, Xun Cheng 1, Charles Martin 1, Dongmei Wu 1, Shuyuan Yao 1, Min-Ju Kim 2, Yang Liu 1
1Research and Development, Stemgent, 2Product Marketing, Stemgent

SC1 functions through dual inhibition of Ras- GAP and ERK1. We tested the function of SC1 in supporting mouse ES cell self-renewal in the absence of LIF and showed that SC1 is able to maintain self-renewal of mouse ES cell cultures.

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Neuroscience

Automated Interactive Video Playback for Studies of Animal Communication
Trisha Butkowski 1, Wei Yan 1, Aaron M. Gray 2, Rongfeng Cui 2, Machteld N. Verzijden 2, Gil G. Rosenthal 2
1Department of Visualization, Texas A&M University (TAMU), 2Department of Biology, Texas A&M University (TAMU)

Video playback is a widely used technique in animal behavior. We created and evaluated a program that applies rules-based, interactive playback of 3-D computer animations in response to real-time, automated data on subject behavior.

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Immunology and Infection

An Analytical Tool-box for Comprehensive Biochemical, Structural and Transcriptome Evaluation of Oral Biofilms Mediated by Mutans Streptococci
Marlise I. Klein 1, Jin Xiao 1,2, Arne Heydorn 3, Hyun Koo 1,4
1Center for Oral Biology, University of Rochester Medical Center, 2State Key Laboratory of Oral Diseases, Sichuan University, 3Department of General Medicine, Glostrup Hospital, Glostrup, Denmark, 4Department of Microbiology and Immunology, University of Rochester Medical Center

Biofilms formed on tooth surfaces are highly complex and exposed to constant innate and exogenous environmental challenges, which modulate their architecture, physiology and transcriptome. We developed a toolbox to examine the composition, structural organization and gene expression of oral biofilms, which can be adapted to other areas of biofilm research.

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Medicine

Collection and Extraction of Saliva DNA for Next Generation Sequencing
Michael R. Goode 1, Soo Yeon Cheong 1, Ning Li 1, William C. Ray 1,2, Christopher W. Bartlett 1,3
1Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital, 2The Interdisciplinary Graduate Program in Biophysics, The Ohio State University, 3Department of Pediatrics, The Ohio State University

DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.

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Bioengineering

Protocol for Biofilm Streamer Formation in a Microfluidic Device with Micro-pillars
Mahtab Hassanpourfard 1, Xiaohui Sun 2, Amin Valiei 1, Partha Mukherjee 3, Thomas Thundat 1, Yang Liu 2, Aloke Kumar 4
1Department of Chemical and Material Engineering, University of Alberta, 2Department of Civil and Environmental Engineering, University of Alberta, 3Department of Mechanical Engineering, Texas A&M University, 4Department of Mechanical Engineering, University of Alberta

Protocols for the study of biofilm formation in a microfluidic device that mimics porous media are discussed. The microfluidic device consists of an array of micro-pillars and biofilm formation by Pseudomonas fluorescens in this device is investigated.

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Medicine

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury
Klaus Zweckberger 1, Yang Liu 1, Jian Wang 1, Nicole Forgione 1, Michael G. Fehlings 1,2,3
1Department of Genetics and Development, Toronto Western Research Institute and Spinal Program, University Health Network, Krembil Neuroscience Centre, 2Department of Surgery, University of Toronto, 3Institute of Medical Sciences, University of Toronto

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

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Bioengineering

Microbiologically Induced Calcite Precipitation Mediated by Sporosarcina pasteurii
Swayamdipta Bhaduri 1, Nandini Debnath 1, Sushanta Mitra 2, Yang Liu 3, Aloke Kumar 1
1Department of Mechanical Engineering, University of Alberta, 2Department of Mechanical Engineering, York University, 3Department of Civil and Environmental Engineering, University of Alberta

Protocols for microbiologically induced calcite precipitation (MICP) using the bacterium Sporosarcina pasteurii are presented here. The precipitated calcium carbonate was characterized through optical microscopy and scanning electron microscopy (SEM). It is also shown that exposure to MICP increases the compressive strength of sponge.

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Medicine

Two Techniques to Create Hypoparathyroid Mice: Parathyroidectomy Using GFP Glands and Diphtheria-Toxin-Mediated Parathyroid Ablation
Ruiye Bi 1,2, Yi Fan 2,3, En Luo 2,4, Quan Yuan 2,4, Michael Mannstadt 1
1Endocrine Unit, Massachusetts General Hospital, Harvard Medical School, 2West China School of Stomatology, Sichuan University, 3Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, 4State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University

Mice with acquired hypoparathyroidism would be useful for studying novel drug therapies for hypoparathyroidism. Two procedures to create such mice are demonstrated. The GFP-PTX mouse is generated by surgical parathyroidectomy guided by green fluorescing parathyroid glands. A second, non-surgical approach is based on parathyroid-specific expression of the diphtheria toxin receptor.

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Education

Facile Preparation of (2Z,4E)-Dienamides by the Olefination of Electron-deficient Alkenes with Allyl Acetate
Liyuan Ding 1, Chunbing Yu 1, Zhenqiang Zhao 2, Feifei Li 1, Jian Zhang 1, Guofu Zhong 1
1College of Materials, Chemistry and Chemical Engineering, Hangzhou Normal University, 2Shijiazhuang Mechanized Infantry Academy

The ruthenium-catalyzed olefination of electron-deficient alkenes with allyl acetate is described here. By using aminocarbonyl as a directing group, this external oxidant-free protocol has high efficiency and good stereo- and regioselectivity, opening a novel synthetic route to (Z,E)-butadiene skeletons.

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Cancer Research

Evaluating In Vitro DNA Damage Using Comet Assay
Yanxin Lu 1,2, Yang Liu 1, Chunzhang Yang 1
1Neuro-Oncology Branch, Center for Cancer Research, National Cancer Institute, 2Basic Medical Science Department, Zunyi Medical College-Zhuhai Campus

The comet assay is an efficient method to detect DNA damage including single and double-stranded DNA breaks. We describe alkaline and neutral comet assays to measure DNA damage in cancer cells to evaluate the therapeutic effect of chemotherapy.

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Engineering

An Available Technique for Preparation of New Cast MnCuNiFeZnAl Alloy with Superior Damping Capacity and High Service Temperature
Dong Li 1,3, Wenbo Liu 1,2, Ning Li 1, Zhenyu Zhong 1, Jiazhen Yan 1, Sanqiang Shi 2
1School of Manufacturing Science and Engineering, Sichuan University, 2Department of Mechanical Engineering, Hong Kong Polytechnic University, 3Department of Mechanical and Electrical Engineering, Chengdu Aeronautic Polytechnic

Here we present a protocol to obtain a novel Mn-Cu-based alloy with excellent comprehensive performances by a high-quality smelting technology and reasonable heat treatment methods.

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Neuroscience

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method
Weiyue Zhang *1, Wei Zhang *1, Ning Dai *1, Chenxia Han 2, Fengzhi Wu 1, Xu Wang 1, Libo Tan 1, Jie Li 1, Feng Li 1, Qingjia Ren 3
1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, 2Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, 3Institute of Tibetan Medicine, Tibetan Traditional Medical College

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

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Immunology and Infection

Design of Cecal Ligation and Puncture and Intranasal Infection Dual Model of Sepsis-Induced Immunosuppression
Zhihan Wang *1,2, Qinqin Pu *1,2, Ping Lin *2,3, Changlong Li 1, Jianxin Jiang 3, Min Wu 2
1West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, 2Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, 3State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Surgery Research, Daping Hospital, Army Medical University

This protocol describes techniques to measure infectious outcomes underlying secondary hospital-acquired infections in the immunosuppressive condition, first by establishing cecal ligation/puncture mice then challenging them with intranasal infection to create a clinically relevant model of immunosuppression sepsis.

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Biology

A Labor-saving and Repeatable Touch-force Signaling Mutant Screen Protocol for the Study of Thigmomorphogenesis of a Model Plant Arabidopsis thaliana
Kai Wang 1, Kayin Law 1, Manhin Leung 1, Waishing Wong 1, Ning Li 1,2
1Division of Life Science, Energy Institute, Institute for the Environment, The Hong Kong University of Science and Technology, 2Shenzhen Research Institute, The Hong Kong University of Science and Technology

A gentle touch-force loading machine is built from human hair brushes, robotic arms and a controller. The hair brushes are driven by robotic arms installed on the machine and move periodically to apply touch-force on plants. The strength of machine-driven hair touches is comparable to that of manually applied touches.

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Behavior

Home-Based Prescribed Pulmonary Exercise in Patients with Stable Chronic Obstructive Pulmonary Disease
Xiaodan Liu 1,2, Peijun Li 3, Jian Li 3, Lu Xiao 1, Ning Li 3, Yufan Lu 3, Zhengrong Wang 3, Jianqing Su 3, Zhenwei Wang 4, Chunlei Shan 1,2, Weibing Wu 3
1School of Rehabilitation Science, Shanghai University of Traditional Chinese Medicine, 2Institute of Rehabilitation Medicine, Shanghai Academy of Traditional Chinese Medicine, 3Department of Sports Medicine, Shanghai University of Sport, 4Department of Respiratory Medicine, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine

Presented here is a protocol to investigate the effects of home-based prescribed pulmonary exercise in stable chronic obstructive pulmonary disease (COPD) patients, which is modified based on traditional Chinese exercises according to dyspnea and limited exercise capacity observed in COPD patients.

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Medicine

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Di Liu *1, Xizhu Xu *2, Yuejin Li 2, Jie Zhang 1, Xiaoyu Zhang 1, Qihuan Li 1, Haifeng Hou 2, Dong Li 2, Wei Wang 1,2,3, Youxin Wang 1
1Beijing Key Laboratory of Clinical Epidemiology, School of Public Health, Capital Medical University, 2School of Public Health, Shandong First Medical University & Shandong Academy of Medical Sciences, 3School of Medical and Health Sciences, Edith Cowan University

Immunoglobulin G (IgG) N-glycan is characterized using hydrophilic interaction chromatography UPLC. In addition, the structure of IgG N-glycan is clearly separated. Presented here is an introduction to this experimental method so that it can be widely used in research settings.

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Behavior

Impact of High-intensity Interval Exercise and Moderate-Intensity Continuous Exercise on the Cardiac Troponin T Level at an Early Stage of Training
Haifeng Zhang 1,2, Jinlei Nie 3, Zhaowei Kong 4, Xiangui Zhu 1, Yang Liu 1, Qingde Shi 3
1Physical Education College, Hebei Normal University, 2Hebei Provincial Key Lab of Measurement and Evaluation in Human Movement and Bio-Information, 3School of Health Sciences and Sports, Macao Polytechnic Institute, 4Faculty of Education, University of Macau

Here, we present protocols of high-intensity interval and moderate-intensity continuous exercise to observe the response of circulating cardiac troponin T (cTnT) concentration to acute exercise over 10 days. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise.

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Neuroscience

Time-lapse Live Imaging and Quantification of Fast Dendritic Branch Dynamics in Developing Drosophila Neurons
Chengyu Sheng 1, Uzma Javed 1, Justin Rosenthal 1, Jun Yin 1, Bo Qin 1, Quan Yuan 1
1National Institute of Neurological Disorders and Stroke, National Institutes of Health

Here, we describe the method we employed to image highly motile dendritic filopodia in a live preparation of the Drosophila larval brain, and the protocol we developed to quantify time-lapse 3D imaging datasets for quantitative assessments of dendrite dynamics in developing neurons.

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Cancer Research

Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane
Moe Ishihara *1, Junhui Hu *1, Xiaoyu Zhang 2, YongHyeon Choi 3, Anthony Wong 4, Celine Cano-Ruiz 5, Rongwei Zhao 6, Ping Tan 7, Jonathan L. Tso 1, Lily Wu 1,8
1Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, 2Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, 3Department of Bioengineering, Hanyang University, 4Department of Ecology and Evolutionary Biology, University of California, Los Angeles, 5Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, 6School of Life Sciences, Beijing Normal University, 7Department of Urology, West China Hospital, Sichuan University, 8Department of Urology, David Geffen School of Medicine, University of California, Los Angeles

Metastatic clear cell renal cell carcinoma is a disease without a comprehensive animal model for thorough preclinical investigation. This protocol illustrates two novel animal models for the disease: the orthotopically implanted mouse model and the chicken chorioallantoic membrane model, both of which demonstrate lung metastasis resembling clinical cases.

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Biology

Fecal Microbiota Transplantation and Detection of Prevalence of IgA-Coated Bacteria in the Gut
Qiuping Zhang 1, Qian Lu 2, Yubin Luo 1
1Department of Rheumatology and Immunology, West China Hospital, Sichuan University, 2Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University

We established an efficient way to deplete intestinal bacteria in three days, and subsequently transplant fecal microbiota via gavage of fecal fluid prepared from fresh or frozen intestinal contents in mice. We also present an optimized method to detect the frequency of IgA-coated bacteria in the gut.

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Engineering

Measurement of Chladni Mode Shapes with an Optical Lever Method
Rong Feng *1, Yan Luo *1, Yixue Dong 1, Mengke Ma 2, Yuqi Wang 2, Jing Zhang 3, Wenjiang Ma 3, Donghuan Liu 4
1School of Materials Science and Engineering, University of Science and Technology Beijing, 2School of Civil and Resource Engineering, University of Science and Technology Beijing, 3Basic Experimental Center for Natural Science, University of Science and Technology Beijing, 4School of Mathematics and Physics, University of Science and Technology Beijing

A simple method of measuring the Chladni mode shape on an elastic plate by the principle of an optical lever is proposed.

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Behavior

Tactile Semiautomatic Passive-Finger Angle Stimulator (TSPAS)
Wu Wang 1, Jiajia Yang 2,4, Yinghua Yu 2,3,4, Qiong Wu 5,2, Satoshi Takahashi 2, Yoshimichi Ejima 2, Jinglong Wu 6,2
1Cognitive Neuroscience Laboratory, Graduate School of Natural Science and Technology, Okayama University, 2Cognitive Neuroscience Laboratory, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, 3Center for Information and Neural Networks, National Institute of Information and Communications Technology, 4Section on Functional Imaging Methods, National Institute of Mental Health, 5School of Education, Suzhou University of Science and Technology, 6Beijing Institute of Technology

Presented is the tactile semiautomated passive-finger angle stimulator TSPAS, a new way to assess tactile spatial acuity and tactile angle discrimination using a computer-controlled tactile stimulus system that applies raised angle stimuli to a subject's passive fingerpad, while controlling for movement speed, distance, and contact duration.

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Immunology and Infection

Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines
Huaping Zheng *1, Linna Gu *1, Zhen Wang *1, Hong Zhou *1, Chen Zhang 1, Xiu Teng 1, Zhonglan Hu 1, Xiaoqiong Wei 1, Xiao Liu 1, Fanlian Zeng 1, Qixiang Zhao 1, Yan Hao 1, Yawen Hu 1, Xiaoyan Wang 1, Jing Hu 1, Jiadong Yu 1, Wenlin Wu 1, Yifan Zhou 1, Kaijun Cui 2, Nongyu Huang 1, Jiong Li 1
1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University and Collaborative Innovation Center for Biotherapy, 2Department of Cardiovascular Medicine, West China Hospital, Sichuan University

This paper presents a method of establishing an in vitro psoriasiform cutaneous inflammatory model at the transcription level using a combination of five cytokines (IL-17A, IL-22, IL-1α, TNF-α, OSM) on HaCaT cell line.

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Biology

Microsurgical Obstruction of Testes Fusion in Spodoptera litura
Xiaolin He *1, Qianqian Ma *1, Baozhu Jian 1, Yucheng Liu 1, Qiong Wu 1, Meixin Chen 1, Qili Feng 1, Ping Zhao 2, Lin Liu 1
1South China Normal University-Shipai Campus: South China Normal University, 2Southwest University

Aluminum foil was microsurgically inserted between the testes of Spodoptera litura to obstruct the fusion of testis. The procedure includes freezing, fixing, disinfection, incision, placing the barrier, suturing, postoperative feeding, and inspection. This approach provides a method to interfere with tissue formation.

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Neuroscience

A Pilot Study on the Repetitive Transcranial Magnetic Stimulation of Aβ and Tau Levels in Rhesus Monkey Cerebrospinal Fluid
Ling-Yi Liao 1,2,3, Ying-Qian Zhang 4, Benson Wui-Man Lau 5, Qiong Wu 6, Zi-Yan Fan 1,3, Qiang Gao 1,3, Zhi-Hui Zhong 6,7
1Department of Rehabilitation Medicine, West China Hospital, Sichuan University, 2Department of Rehabilitation Medicine, Daping Hospital, Army Medical University, 3Key Laboratory of Rehabilitation Medicine in Sichuan Province, West China Hospital, Sichuan University, 4Department of physiology, Southwest Medical University, 5Department of Rehabilitation Sciences, The Hong Kong Polytechnic University, 6Laboratory of Nonhuman Primate Disease Modeling Research, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, 7Sichuan Kangcheng Biotech Co., Inc.

Here, we describe the procedure for a pilot study to explore the effect of repetitive transcranial magnetic stimulation with different frequencies (1 Hz/20 Hz/40 Hz) on Aβ and tau metabolism in rhesus monkey cerebrospinal fluid.

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Immunology and Infection

Analysis of Granulocyte-Macrophage Colony-Stimulating Factor-Producing T Helper Cells in a Mouse Model of Contact Hypersensitivity
Xiu Teng 1,2, Li-Ping Li 1,2, Meng Tang 1,2, Hui-Fang Li 3, Xiang-Yi Ren 3, Ling Jiao 1,2, Sha-Sha Wu 1,2, Da-Chao Mou 1,2, Zhi-Yong Miao 1,2
1Laboratory of Human Disease and Immunotherapies, West China Hospital, Sichuan University, 2Institute of Immunology and Inflammation, Frontiers Science Center for Disease-Related Molecular Network, West China Hospital, Sichuan University, 3Core Facilities of West China Hospital, Sichuan University

Here, we present a simple and standardized method of analyzing the granulocyte-macrophage colony-stimulating factor-producing T helper subset in vivo.

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Biochemistry

A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors
Lin Cheng *1, Lantian Su *1, Xiaowen Tian *1, Fan Xia 2, Chang Zhao 1, Wei Yan 1, Zhenhua Shao 1
1Division of Nephrology and Kidney Research Institute, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, 2Department of Neurosurgery, West China Hospital, Sichuan University

S1P exerts its diverse physiological effects through the S1P receptors (S1PRs) subfamily. Here, a pipeline is described to expound on the structures and function of S1PRs.

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Cancer Research

Measurement of the Compressibility of Cell and Nucleus Based on Acoustofluidic Microdevice
Qibin Fu 1, Yan Zhang 1, Tuchen Huang 1, Yang Liu 1
1Sino-French Institute of Nuclear Engineering and Technology, Sun Yat-sen University

Here a protocol is presented to build a fast and non-destructive system for measuring cell or nucleus compressibility based on acoustofluidic microdevice. Changes in mechanical properties of tumor cells after epithelial-mesenchymal transition or ionizing radiation were investigated, demonstrating the application prospect of this method in scientific research and clinical practice.

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Medicine

Technical Applications of Microelectrode Array and Patch Clamp Recordings on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Shane Rui Zhao 1,2, Gema Mondéjar-Parreño 1,2, Dong Li 1,2, Mengcheng Shen 1,2, Joseph C. Wu 1,2,3
1Stanford Cardiovascular Institute, Stanford University School of Medicine, 2Division of Cardiovascular Medicine, Department of Medicine, Stanford University School of Medicine, 3Department of Radiology, Stanford University School of Medicine

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have emerged as a promising in vitro model for drug-induced cardiotoxicity screening and disease modeling. Here, we detail a protocol for measuring the contractility and electrophysiology of hiPSC-CMs.

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Medicine

Transoral Endoscopic Thyroidectomy Vestibular Approach for Thyroid Lobectomy
Zhujuan Wu *1, Ye Tian *2, Ziwei Chen 1, Anping Su 2
1West China School of Medicine, Sichuan University, 2Sichuan University West China Hospital

Here, we present a protocol to describe the methodology for the transoral endoscopic thyroidectomy vestibular approach in detail.

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Medicine

A Personalized 3D-Printed Model for Preoperative Evaluation in Thyroid Surgery
Pengyu Li *1, YIFei Chen *2, Wanjun Zhao 1, Zhimian Huang 3, Jingqiang Zhu 1
1Department of Thyroid Surgery, West China Hospital, Sichuan University, 2West China School/Hospital of Stomatology, Sichuan University, 3Encomomic School of New York University Shanghai

Here, a new method of establishing a personalized 3D-printed model for preoperative evaluation of thyroid surgery is proposed. It is conducive to preoperative discussion, reducing the difficulty of thyroid surgery.

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Cancer Research

Spontaneous Murine Model of Anaplastic Thyroid Cancer
Huayun Yan 1, Yingfang Ma 1, Xinyue Zhou 2, Yushuang He 3, Yang Liu 2, Carlos Caulin 4, Leiming Wang 5, Heng Xu 1,6, Han Luo 2,6
1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center, 2Division of Thyroid Surgery, Laboratory of Thyroid and Parathyroid Disease, Frontiers Science Center for Disease-Related Molecular Network, Department of General Surgery, West China Hospital, Sichuan University, 3Department of Ultrasound, West China Hospital of Sichuan University, Sichuan University, 4Department of Otolaryngology-Head & Neck Surgery and University of Arizona Cancer Center, University of Arizona, 5Center for Translational Medicine, Shenzhen Bay Laboratory, 6Division of Laboratory Medicine/Research Centre of Clinical Laboratory Medicine, West China Hospital, Sichuan University

Here, we present a standard pipeline to obtain murine ATC tumors by spontaneous genetically engineered mouse models. Further, we present tumor dynamics and pathological information about the primary and metastasized lesions. This model will help researchers to understand tumorigenesis and facilitate drug discoveries.

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Medicine

Generation of Hypoparathyroid Rats via Carbon-Nanoparticle-Assisted Parathyroidectomy
Yiming Song *1, Peiran Li *2, Ping Lyu 1, Yanshen Yu 1, Xinyu Chen 1, Chen Cui 3, Ruiye Bi 2, Yi Fan 1
1State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, 2State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Department of Orthognathic and TMJ Surgery, West China Hospital of Stomatology, 3Guangdong Province Key Laboratory of Stomatology, Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-Sen University

An animal model of acquired hypoparathyroidism (HypoPT) is crucial to understanding how HypoPT affects mineral ion homeostasis and to verify the effectiveness of novel treatments. Here, a technique is presented to generate an acquired hypoparathyroidism (AHypoPT) rat model by parathyroidectomy (PTX) using carbon nanoparticles.

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Medicine

Gasless Endoscopic Thyroidectomy via the Trans-Axillary Approach
Xun Zheng 1, Xiaofei Wang 1, Tao Wei 1
1Division of Thyroid Surgery, Department of General Surgery, West China Hospital, Sichuan University

This protocol presents the procedure for conducting thyroidectomy using the gasless endoscopic thyroidectomy trans-axillary approach (GETTA).

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Cancer Research

High-Resolution Ultrasonography for the Analysis of Orthotopic ATC Tumors in a Genetically Engineered Mouse Model
Yushuang He 1, Yan Luo 1, Zhihui Li 2, Han Luo 2, Huayun Yan 3, Yingfang Ma 3, Xinyue Zhou 2
1Department of Ultrasound, West China Hospital, Sichuan University, 2Department of Thyroid and Parathyroid Surgery, Laboratory of Thyroid and Parathyroid Disease, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, 3State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center

The present protocol describes high-frequency ultrasonography for visualizing the entire mouse thyroid gland and monitoring the growth of anaplastic thyroid carcinoma.

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Cancer Research

Establishment and Characterization of Patient-Derived Xenograft Models of Anaplastic Thyroid Carcinoma and Head and Neck Squamous Cell Carcinoma
Maodi Wu 1,3, Yang Liu 2, Yi Zhao 3, Yuyao Zhang 4, Liansi Huang 3, Qijun Du 3, Ting Zhang 1,3, Zhihui Zhong 1,3, Han Luo 2, Kai Xiao 1,3
1Precision Medicine Research Center, Sichuan Provincial Key Laboratory of Precision Medicine, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, 2Department of Thyroid Surgery, West China Hospital, Sichuan University, 3Sichuan Kangcheng Biotech Co., 4Sichuan Cancer Hospital

The present protocol establishes and characterizes a patient-derived xenograft (PDX) model of anaplastic thyroid carcinoma (ATC) and head and neck squamous cell carcinoma (HNSCC), as PDX models are rapidly becoming the standard in the field of translational oncology.

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Engineering

A Teleoperated Robotic System-Assisted Percutaneous Transiliac-Transsacral Screw Fixation Technique
Binbin Liu 1, Fei Xu 1, Yang Liu 1, Tianqi Wang 2, Yajing Cao 2, Zehang Zheng 1, Hanqing Xu 1, Chuang Huang 1, Zhengqiang Luo 1
1Department of Orthopedics, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, 2Wuhan United Imaging Healthcare Surgical Technology Co. Ltd.

Teleoperated robotic system-assisted percutaneous transiliac-transsacral screw fixation is a feasible technique. Screw channels can be implemented with high accuracy owing to the excellent freedom of movement and stability of the robotic arms.

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Biology

The Establishment of a Murine Mandibular Molar Extraction Socket Healing Model
Changhao Yu 1,2, Fanyuan Yu 1,2, Feifei Li 1,2, Ling Ye 1,2
1State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, 2Department of Endodontics, West China Stomatology Hospital, Sichuan University

This protocol demonstrates step-by-step details of how to extract the mandibular first molar in the mouse. It provides an alternative method for researchers focusing on jawbone healing and regeneration.

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Biology

Unique Approach for Isolating Rat Bone Marrow Neutrophils with Comparable Capacity
Xiangfeng Gong *1, Yutao Sun *1, Xiaoxin Zhang 2,3, Zhenghua Xiao 1,4, Li He 1,4, Chaoyi Qin 1,4
1Department of Cardiovascular Surgery, West China Hospital, Sichuan University, 2Department of Integrated Traditional Chinese and Western Medicine, Sichuan Provincial Pancreatitis Centre, West China Hospital, Sichuan University, 3West China-Liverpool Biomedical Research Centre, West China Hospital, Sichuan University, 4Cardiovascular Surgery Research Laboratory, West China Hospital, Sichuan University

This research outlines two techniques for isolating abundant neutrophil extracellular traps (NETs) from rat bone marrow. One method combines a commercial neutrophil isolation kit with density gradient centrifugation, while the other employs only density gradient centrifugation. Both approaches yield functional NETs surpassing those from peripheral blood neutrophils.

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Medicine

Establishment of an Oronasal Fistula Mice Model
Jiali Chen 1, Jiayi Yin 1, Shiming Zhang 1, Shiyong Zhuang 2, Renjie Yang 3, Yichen Xu 4, Qian Zheng 1, Bing Shi 1, Hanyao Huang 1
1State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases and Department of Oral Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, 2State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases and Department of Head and Neck Oncology, West China Hospital of Stomatology, Sichuan University, 3State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases and Eastern Clinic, West China Hospital of Stomatology, Sichuan University, 4State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases and Department of Oral Prosthodontics, West China Hospital of Stomatology, Sichuan University

This article outlines a step-by-step procedure for establishing a mice model with an oronasal fistula. The oronasal fistula was created by employing heated ophthalmologic cautery to damage the midline portion of the hard palate, resulting in the formation of an opening between the oral and nasal cavities.

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Biology

Freezing Injury in Mouse Masseter Muscle to Establish an Orofacial Muscle Fibrosis Model
Xu Cheng *1, Yixuan Huang *2, Yanan Li 2, Jingtao Li 2, Yan Wang 2
1State Key Laboratory of Oral Diseases, West China School of Stomatology, Sichuan University, 2Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University

The goal of this protocol is to establish an orofacial muscle fibrosis model. Comparison of the histology between mice masseter and tibialis anterior muscle after freezing injury confirmed masseter muscle fibrosis. This model will facilitate further investigation into the mechanism underlying orofacial muscle fibrosis.

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Cancer Research

Multiplex Immunohistochemistry Staining for Paraffin-embedded Lung Cancer Tissue
Ying Yang 1, Hong Huang 1, Li Li 2, Yongfeng Yang 1
1Department of Respiratory and Critical Care Medicine, Center of Precision Medicine, West China Hospital, Sichuan University, 2Institute of Clinical Pathology, West China Hospital, Sichuan University

This experimental protocol describes and optimizes a multiplex immunohistochemistry (IHC) staining method, mainly by optimizing single-channel antibody incubation conditions and adjusting the settings of antibodies and channels to solve the problems of autofluorescence and channel crosstalk in lung cancer tissues of clinical origin.

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Biology

An Efficient and Fast Method for Labeling and Analyzing Mouse Glomeruli
Lin Bai 1,2, Yaping Wu 2, Wenshu Dai 1, Qiuxiao Shi 2, Lei Wu 2, Jie Zhang 1,2, Lily Zheng 3
1Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, 2Core Facility of West China Hospital, Sichuan University, 3Regenerative Medicine Research Center, West China Hospital, Sichuan University

This study presents an easy-to-use, complete, and simple set of methods to label and analyze glomeruli from CUBIC-cleared mouse kidneys. Data such as glomerulus number and volume can be obtained easily and reliably using fluorescein isothiocyanate (FITC)-Dextran, light sheet fluorescence microscopy (LSFM), or common confocal microscopy and software such as Imaris.

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Medicine

Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research
Yawen Tang 1,2, Changhao Yu 1,2, Feifei Li 1, Haisheng Wang 1, Ling Ye 1,2
1State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, 2Department of Endodontics, West China Hospital of Stomatology, Sichuan University

This article presents a streamlined protocol for establishing a pulpitis model in mice using an innovative mouth-gag, followed by subsequent histological analysis.

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Medicine

The Establishment of a Murine Maxillary Orthodontic Model
Jianxin Liu 1,2, Changhao Yu 1,2, Feifei Li 1,2
1State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, 2Department of Endodontics, West China Stomatology Hospital, Sichuan University

Here we demonstrate step-by-step a manageable, orthodontic tooth movement protocol operated on a murine maxillary model. With the explicit explanation of each step and visual demonstration, researchers can master this model and apply it to their experimental needs with a few modifications.

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Medicine

The Establishment of Calvarial Suture-Bony Composite Defects in Rats: A Standardized Model for Suture-Regenerative Therapy Investigation
Jiayi Wu 1,2, Changhao Yu 1,2, Chuyi Han 1, Feifei Li 1, Haisheng Wang 1, Bei Yin 1,2
1State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, 2Department of Endodontics, West China Hospital of Stomatology, Sichuan University

We describe the detailed surgical procedures of calvarial suture-bony composite defects in rats, alongside the investigations into the short-term and long-term prognoses of the model. We aim to construct a standardized model for developing suture-regenerative therapies.

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Developmental Biology

Rapid Isolation of Stage I Oocytes in Zebrafish Devoid of Granulosa Cells
Qianwen Zheng 1, Xiaolong Xie 1, Yao Li 1, Chengbo Ai 1, Siyu Pu 1, Jing Chen 1
1Department of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital / West China School of Medicine, Sichuan University

This protocol describes a modified procedure for rapidly isolating clean stage I oocytes in zebrafish devoid of granulosa cells, thereby providing a convenient method for oocyte-specific research.

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Medicine

A Mouse Abdominal Aortic Aneurysm Model by Periadventitial Calcium Chloride and Elastase Infiltration
Jinting Ge *1, Chengxin Weng *1, Xiaoling Zhang 2, Jichun Zhao 1, Hua Zhang 2, Ding Yuan 1, Tiehao Wang 1
1Division of Vascular Surgery, Department of General Surgery, West China Hospital, Sichuan University, 2Key Laboratory of Birth Defects and Related Diseases of Women and Children of MOE, State Key Laboratory of Biotherapy, West China Second University Hospital, Sichuan University

This manuscript presents a protocol for establishing a mouse abdominal aortic aneurysm model using calcium chloride and elastase, combining the advantages of previous modeling methods. This model can be utilized to investigate the pathophysiological mechanisms underlying abdominal aortic aneurysms.

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Cancer Research

Effective Detection of Hoechst Side Population Cells by Flow Cytometry
Fangfang Wang 1,2, Xinyu Zhai 1, Tingting Guo 3, Hengyi Xiao 2, Jingcao Huang 1
1Institute of Hematology, Department of Hematology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, 2The Lab of Aging Research, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, 3Precision Medicine Research Laboratory, West China Hospital, Sichuan University

Here, we show that high-power 375 nm and 405 nm lasers can effectively excite Hoechst 33342 and serve as a viable alternative to the 355 nm laser for side population (SP) cell detection, thereby expanding the range of available lasers in flow cytometry applications.

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