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73 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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Medicine

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)
Yana Yunusova 1,2, Jordan R. Green 3, Jun Wang 3, Gary Pattee 4, Lorne Zinman 2,5
1Department of Speech-Language Pathology, University of Toronto, 2ALS/ MN Clinic, Sunnybrook Health Science Centre, 3Department of Special Education and Communication Disorders, University of Nebraska-Lincoln, 4Department of Neurology, Munroe-Meyer Institute, University of Nebraska Medical Center, 5Department of Neurology, University of Toronto

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

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Medicine

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD
Min Jia 1, Fei Meng 2, Stanley E. Smerin 1, Guoqiang Xing 1, Lei Zhang 1, David M. Su 2, David Benedek 1, Robert Ursano 1, Yan A. Su 2, He Li 1
1Department of Psychiatry, Center for the Study of Traumatic Stress, Uniformed Services University of the Health Sciences, Bethesda, Maryland, 2Department of Gene and Protein Biomarkers, GenProMarkers, Inc.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

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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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Developmental Biology

Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells
Ieng Lam Lei 1, Lei Bu 2, Zhong Wang 1
1Cardiac Surgery, University of Michigan, 2Leon H Charney Division of Cardiology, New York University School of Medicine

In this protocol, derivation of cardiac progenitor cells from both mouse and human embryonic stem cells will be illustrated. A major strategy in this protocol is to enrich cardiac progenitor cells with flow cytometry using fluorescent reporters engineered into the embryonic stem cell lines.

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Bioengineering

Porous Silicon Microparticles for Delivery of siRNA Therapeutics
Jianliang Shen *1,2, Xiaoyan Wu *1,3, Yeonju Lee 1, Joy Wolfram 1,4, Zhizhou Yang 1, Zong-Wan Mao 2, Mauro Ferrari 1,5, Haifa Shen 1,6
1Department of Nanomedicine, Houston Methodist Research Institute, 2MOE Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry and Chemical Engineering, Sun Yat-sen University, 3Pediatrics Department of Union Hospital, Huazhong University of Science and Technology, 4CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience & Technology of China, 5Department of Medicine, Weill Cornell Medical College, 6Department of Cell and Developmental Biology, Weill Cornell Medical College

Delivery remains the main challenge for the therapeutic implementation of small interfering RNA (siRNA). This protocol involves the use of a multifunctional and biocompatible siRNA delivery platform, consisting of arginine and polyethylenimine grafted porous silicon microparticles.

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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

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
Yunfeng Chen *1, Baoyu Liu *2, Lining Ju *3, Jinsung Hong *1, Qinghua Ji 4,5, Wei Chen 6, Cheng Zhu 1
1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 3Charles Perkins Centre, The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University

We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte.

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Bioengineering

Fabrication of a Bioactive, PCL-based "Self-fitting" Shape Memory Polymer Scaffold
Lindsay N. Nail 1, Dawei Zhang 2,3, Jessica L. Reinhard 1, Melissa A. Grunlan 1,2
1Department of Biomedical Engineering, Texas A&M University, 2Department of Material Science and Engineering, Texas A&M University, 3Institute of Advanced Materials and Technology, University of Science & Technology Beijing

Scaffolds capable of fitting within cranio-maxillofacial (CMF) bone defects while exhibiting osteoconductivity and bioactivity are of interest. This protocol describes the preparation of a shape memory scaffold based on polycaprolactone diacrylate (PCL-DA) using a solvent-casting particulate-leaching (SCPL) method employing a fused salt template and application of a bioactive polydopamine coating.

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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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Chemistry

Flow-pattern Guided Fabrication of High-density Barcode Antibody Microarray
Lisa S. Ramirez 1,2, Jun Wang 1,2
1Department of Chemistry, University at Albany, State University of New York, 2Multiplex Biotechnology Laboratory, Cancer Research Center

This protocol outlines the fabrication of a large-scale, multiplexed two-dimensional DNA or antibody array, with potential applications in cell signaling studies and biomarker detection.

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Biochemistry

Real-time Tracking of DNA Fragment Separation by Smartphone
Chunxian Tao 1, Bo Yang 1, Zhenqing Li 1, Dawei Zhang 1, Yoshinori Yamaguchi 1,2,3
1Shanghai Key Lab of Modern Optical System, University of Shanghai for Science and Technology, 2Department of Applied Physics, Graduate School of Engineering, Osaka University, 3East China University of Science and Technology, Department of Physics Faculty of Science

Traditional slab gel electrophoresis (SGE) experiments require a complicated apparatus and high chemical consumption. This work presents a protocol that describes a low-cost method to separate DNA fragments within a short timeframe.

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Environment

Preparation of Authigenic Pyrite from Methane-bearing Sediments for In Situ Sulfur Isotope Analysis Using SIMS
Zhiyong Lin 1,3, Xiaoming Sun 1,2,3,4, Jörn Peckmann 5, Yang Lu 2,3, Harald Strauss 6, Li Xu 2,3, Hongfeng Lu 7, Barbara M.A. Teichert 6
1School of Earth Sciences and Engineering, Sun Yat-sen University, 2School of Marine Sciences, Sun Yat-sen University, 3Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, 4South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, 5Institut für Geologie, Universität Hamburg, 6Institut für Geologie und Paläontologie, Westfälische Wilhelms-Universität Münster, 7Guangzhou Marine Geological Survey

Analyses of the sulfur isotopic composition (δ34S) of pyrite from methane-bearing sediments have typically focused on bulk samples. Here, we applied secondary ion mass spectroscopy to analyze the δ34S values of various pyrite generations to understand the diagenetic history of pyritization.

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JoVE Journal

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
Zackary N. Scholl 1, Qing Li 1, Eric Josephs 1, Dimitra Apostolidou 1, Piotr E. Marszalek 1
1Mechanical Engineering and Materials Science, Duke University

We describe the detailed procedures and strategies to measure the mechanical properties and mechanical unfolding pathways of single protein molecules using an atomic force microscope. We also show representative results as a reference for selection and justification of good single protein molecule recordings.

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Behavior

Bouncing Ball with a Uniformly Varying Velocity in a Metronome Synchronization Task
Yingyu Huang *1, Li Gu *1, Junkai Yang *1, Xiang Wu 1
1Department of Psychology, Sun Yat-Sen University

The purpose of this protocol is to introduce the application of a bouncing ball with a uniformly varying velocity in a metronome synchronization task.

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Medicine

Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening
Hui Jiang *1, Yicun Wang *1, Zhantao Deng *1,2,3, Jiewen Jin 2,3, Jia Meng 1, Shuo Chen 1, Jun Wang 1, Yang Qiu 1, Ting Guo 1, Jianning Zhao 1
1Department of Orthopedics, Jinling Hospital, School of Medicine, Nanjing University, 2Center for Translational Medicine, Nanjing University Medical School, 3Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School

This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening.

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Medicine

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients
Xiang Wu 1,2, Guoyi Gao 1,2, Junfeng Feng 1,2, Qing Mao 1,2, Jiyao Jiang 1,2
1Department of Neurosurgery, Renji Hospital, Shanghai Jiaotong University School of Medicine, 2Shanghai Institute of Head Trauma

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

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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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Developmental Biology

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Qinggang Dai *1, Yujiao Han *2, Furong Xie *1, Xuhui Ma 3, Zhan Xu 2, Xiao Liu 1, Weiguo Zou 2, Jun Wang 1
1Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, 2State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 3Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine

This manuscript describes a protocol to isolate and culture osteoclasts in vitro from mouse bone marrow, and to study the role of the mammalian/mechanistic target of rapamycin complex 1 in osteoclast formation.

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Chemistry

Fabrication of Spherical and Worm-shaped Micellar Nanocrystals by Combining Electrospray, Self-assembly, and Solvent-based Structure Control
Xinyi Ding 1,2,3, Yuxiang Sun 1,2,3, Yanming Chen 1,2,3, Wanchuan Ding 1,2,3, Steven Emory 1,2,3,4, Tianhao Li 1,2,3, Zixing Xu 1,2,3, Ning Han 1,2,3, Jun Wang 1,2,3, Gang Ruan 1,2,3
1Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University, 2Institute of Materials Engineering, College of Engineering and Applied Sciences, Nanjing University, 3Collaborative Innovation Center of Chemistry for Life Sciences, Nanjing University, 4Department of Chemistry, Western Washington University

The present work describes a method to fabricate micellar nanocrystals, an emerging major class of nanobiomaterials. This method combines top-down electrospray, bottom-up self-assembly, and solvent-based structure control. The fabrication method is largely continuous, can produce high quality products, and possesses an inexpensive means of structure control.

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Medicine

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate (HoLEP).
Huan Xu *1, Xiang Wan *1, Meng Gu 1, Yanbo Chen 1, Qiling Shi 1, Qi Chen 1, Zhong Wang 1
1Department of Urology, Shanghai 9th People's Hospital, Shanghai Jiaotong University School of Medicine

Here we present a safe and efficient protocol, holmium laser enucleation of the prostate, to treat benign prostatic hyperplasia.

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Engineering

Determination of the Excitation and Coupling Rates Between Light Emitters and Surface Plasmon Polaritons
Zhaolong Cao 1,2, Min Lin 2, Daniel Ong 2
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.

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Cancer Research

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro
Yupeng Yao 1, Weifan Lin 1, Yan Zhang 1
1Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University

A method is shown here for the preparation of the tongue extracellular matrix (TEM) with efficient decellularization. The TEM can be used as functional scaffolds for the reconstruction of a tongue squamous cell carcinoma (TSCC) model under static or stirred culture conditions.

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Biology

Implantation of Electrospun Vascular Grafts with Optimized Structure in a Rat Model
Kang Qin 1, Yifan Wu 1, Yiwa Pan 1, Kai Wang 1, Deling Kong 1, Qiang Zhao 1
1State Key Laboratory of Medicinal Chemical Biology, Key Laboratory of Bioactive Materials (Ministry of Education), College of Life Sciences, Nankai University

Here, we present a modified electrospinning method to fabricate PCL vascular grafts with thick fibers and large pores, and describe a protocol to evaluate the in vivo performance in a rat model of abdominal aorta replacement.

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Medicine

Adeno-Associated Virus-Mediated Delivery of CRISPR for Cardiac Gene Editing in Mice
Li Xu 1, Yandi Gao 1, Yeh Siang Lau 1, Renzhi Han 1
1Department of Surgery, Davis Heart and Lung Research Institute, Biomedical Sciences Graduate Program, Biophysics Graduate Program, Ohio State University Wexner Medical Center

Here we provide a detailed protocol to carry out in vivo cardiac gene editing in mice using recombinant Adeno-Associated Virus(rAAV)-mediated delivery of CRISPR. This protocol offers a promising therapeutic strategy to treat dystrophic cardiomyopathy in Duchenne muscular dystrophy and can be used to generate cardiac-specific knockout in postnatal mice.

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Cancer Research

A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
Yanlan Wang 1,2, Jiayu Liu 1,2, Haitao Pan 1,2, Jieyu Xing 1,2, Xiaoqiong Wu 1,2, Qing Li 1,2, Zhong Wang 1,2
1School of Pharmaceutical Sciences, Sun Yat-Sen University, 2Center for Cellular & Structural Biology, Sun Yat-Sen University

Here, we present a protocol to produce a bispecific antibody GPC3-S-Fab in Escherichia coli. The purified GPC3-S-Fab has potent cytotoxicity against GPC3 positive liver cancer cells.

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Immunohistochemical Staining of CD8 Protein in Tissue Sections from Patients with Tumors
Shiqiang Zhang *1, Jun Wang *2, Juan Fang *3, Binyuan Yan 1, Haiyun Xiong 1, Fu Shi 1, Guolong Liao 1, Xiaofang Lu 4, Jun Pang 1
1Department of Urology, the Seventh Affiliated Hospital, Sun Yat-sen University, 2Zhongshan School of Medicine, Sun Yat-sen University, 3Guanghua School of Stomatology, Sun Yat-sen University, 4Department of Pathology, the Seventh Affiliated Hospital, Sun Yat-sen University

The present study presents a protocol for immunohistochemistry (IHC) that can image specific antigens located on tissue sections and determine the relative expression level of a target protein or the distribution of target cells.

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Environment

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens
Cai Wang 1, Huifang Wang 1, Tao Ma 1,2, Qiang Xiao 3, Panrong Cao 4, Xuan Chen 5, Hongpeng Xiong 1, Wenquan Qin 1, Zhaohui Sun 1, Xiujun Wen 1
1Guangdong Key Laboratory for Innovation Development and Utilization of Forest Plant Germplasm, College of Forestry and Landscape Architecture, South China Agricultural University, 2College of Agriculture, South China Agricultural University, 3Tea Research Institute, Chinese Academy of Agricultural Sciences, 4College of Horticulture, South China Agricultural University, 5College of Coast and Environment, Louisiana State University

Here, we present a protocol to investigate the pupation preference of mature larvae of Ectropis grisescens in response to soil factors (e.g., substrate type and moisture content) using choice bioassays. We also present a protocol of no-choice bioassays to determine the factors that affect the pupation behaviors and survivorship of E. grisescens.

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Engineering

Measurement of Dynamic Force Acted on Water Strider Leg Jumping Upward by the PVDF Film Sensor
Lele Zhang 1, Meirong Zhao 1, Zhiyi Wang 1, Yunxi Li 1, Yinguo Huang 1, Yelong Zheng 1
1State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University

The protocol here is dedicated to investigating the free and quick maneuvering of water strider on water surface. The protocol includes observing the microstructure of legs and measuring the adhesion force when departing from water surface at different speeds.

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Genetics

Culturing and Manipulation of O9-1 Neural Crest Cells
Bao H. Nguyen 1, Mamoru Ishii 2, Robert E. Maxson 2, Jun Wang 1
1Molecular Physiology and Biophysics, Baylor College of Medicine, 2Department of Biochemistry and Molecular Biology, University of Southern California

O9-1 is a multipotent mouse neural crest cell line. Here we describe detailed step-by-step protocols for culturing O9-1 cells, differentiating O9-1 cells into specific cell types, and genetically manipulating O9-1 cells by using siRNA-mediated knockdown or CRISPR-Cas9 genome editing.

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Chemistry

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy
Zhong Wang 1, Benjamin F. Jeffries 1, Hannah R. Felstead 1, Neil J. Wells 1, Elisabetta Chiarparin 2, Bruno Linclau 1
1Chemistry, University of Southampton, 2Medicinal Chemistry, Oncology, IMED Biotech Unit, AstraZeneca

A novel and straightforward variation of the shake-flask method was developed for accurate lipophilicity measurement of fluorinated compounds by 19F NMR spectroscopy.

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Chemistry

Efficient Construction of Drug-like Bispirocyclic Scaffolds Via Organocatalytic Cycloadditions of α-Imino γ-Lactones and Alkylidene Pyrazolones
Yani Zhou 1, Nannan Chen 2, Yaping Cheng 2, Xiaoqing Cai 2
1School of Basic Medical Sciences, Lanzhou University, 2School of Pharmaceutical Sciences, Sun Yat-sen University

Enantiomerically enriched bispiro[γ-butyrolactone-pyrrolidin-4,4'-pyrazolone] skeletons are asymmetrically synthesized through a simple organocatalytic 1,3-dipolar cycloaddition reaction.

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Cancer Research

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Yu Zhang 1, Yupeng Yao 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The bone extracellular matrix (BEM) model for osteosarcoma (OS) is well established and shown here. It can be used as a suitable scaffold for mimicking primary tumor growth in vitro and providing an ideal model for studying the histologic and cytogenic heterogeneity of OS.

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JoVE Core

Measuring the Interaction Force Between a Droplet and a Super-hydrophobic Substrate by the Optical Lever Method
Shuya Zhuang 1, Meirong Zhao 1, Zhiyi Wang 1, Lele Zhang 1, Yinguo Huang 1, Yelong Zheng 1
1State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University

The protocol aims to investigate the interaction between droplets and super-hydrophobic substrates in the air. This includes calibrating the measurement system and measuring the interaction force at super-hydrophobic substrates with different grid fractions.

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Behavior

Application of 3D Printing in the Construction of Burr Hole Ring for Deep Brain Stimulation Implants
Jiazhi Chen *1,2, Xinyu Chen *3, Siyuan Lv 2, Yuzhen Zhang 1, Hao Long 1, Kaijun Yang 1, Songtao Qi 1, Wangming Zhang 2, Jun Wang 1
1Department of Neurosurgery, Nanfang Hospital, Southern Medical University, 2Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, 3Faculty of Brain Sciences, University of College London

Here, we present a protocol to demonstrate 3D printing in the construction of deep brain stimulation implants.

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Biology

Isolation of Intact Eyeball to Obtain Integral Ocular Surface Tissue for Histological Examination and Immunohistochemistry
Dianlei Guo 1, Chunqiao Liu 1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University

This protocol describes a method for the isolation of the mouse eyeball with eyelid, ocular surface, anterior and posterior segments in relatively intact position.

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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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Cancer Research

Multidimensional Coculture System to Model Lung Squamous Carcinoma Progression
Shuang Chen 1, Andreas Giannakou 1, Jonathon Golas 1, Kenneth G. Geles 1
1Oncology R&D group, Pfizer Worldwide Research and Development

An in vitro model system was developed to capture tissue architectural changes during lung squamous carcinoma (LUSC) progression in a 3-dimensional (3D) co-culture with cancer-associated fibroblasts (CAFs). This organoid system provides a unique platform to investigate the roles of diverse tumor cell-intrinsic and extrinsic changes that modulate the tumor phenotype.

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Chemistry

Cercosporin-Photocatalyzed [4+1]- and [4+2]-Annulations of Azoalkenes Under Mild Conditions
Philippe Icyishaka *1, Chenxing Li *1, Liushen Lu 1, Wenhao Bao 1, Jia Li 1, Yan Zhang 2, Yijian Rao 1
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 2School of Pharmaceutical Science, Jiangnan University

New routes for the synthesis of nitrogen-containing heterocycles utilizing cercosporin as a metal-free photocatalyst were developed.

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Immunology and Infection

Modeling Hepatitis B Virus Infection in Non-Hepatic 293T-NE-3NRs Cells
Xiaoyue Sun *1,2,3, Xiaoqiang Yang *1,2,3,4, Chui Zeng 1,2,3, Fayed Attia Koutb Megahed 1,2,3,5, Qi Zhou 1,2,3, Tingdang Liu 1,2,3, Waqas Iqbal 1,2,3, Pingnan Sun 1,2,3, Xiaoling Zhou 1,2,3
1Stem Cell Research Center, Shantou University Medical College, 2The Center for Reproductive Medicine, Shantou University Medical College, 3Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, 4Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, 5Department of Nucleic Acid Researches, Genetic Engineering and Biotechnology Research Institute, General Autority - City of Scientific Researches and Technological Applications

This manuscript describes a detailed protocol for Hepatitis B virus (HBV) infection in novel engineered 293T cells (293T-NE-3NRs, expressing human NTCP, HNF4α, RXRα and PPARα) and traditional hepatic cells (HepG2-NE, expressing human NTCP).

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Medicine

Avoiding Ischemia Reperfusion Injury in Liver Transplantation
Yixi Zhang *1,2,3, Changjun Huang *1,2,3, Weiqiang Ju *1,2,3, Qiang Zhao 1,2,3, Maogen Chen 1,2,3, Linhe Wang 1,2,3, Dongping Wang 1,2,3, Linwei Wu 1,2,3, Zebin Zhu 4, Shanzhou Huang 5, Zhiheng Zhang 1,2,3, Caihui Zhu 1,2,3, Rongxing Xie 1,2,3, Yinghua Chen 1,2,3, Anbin Hu 1,2,3, Xiaofeng Zhu 1,2,3, Zhiyong Guo 1,2,3, Xiaoshun He 1,2,3
1Organ Transplant Center, The First Affiliated Hospital, Sun Yat-sen University, 2Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology, 3Guangdong Provincial International Cooperation Base of Science and Technology, 4Organ Transplant Center, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, 5Department of General Surgery, Guangdong General Hospital, Guangdong Academy of Medical Sciences

Presented here is a protocol to provide a step-by-step ischemia-free liver transplantation protocol under ex situ normothermic machine perfusion (37 °C) of human livers from donors to recipients.

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Biology

Monitoring Dynamic Growth of Retinal Vessels in Oxygen-Induced Retinopathy Mouse Model
Yuan Ma 1, Tao Li 1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University

This protocol describes a detailed method for the preparation and immunofluorescence staining of mice retinal flat mounts and analysis. The use of fluorescein fundus angiography (FFA) for mice pups and image processing are described in detail as well.

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Cancer Research

Tumor Transplantation for Assessing the Dynamics of Tumor-Infiltrating CD8+ T Cells in Mice
Lisha Wang 1, Zhiming Wang 1, Junyi Guo 2, Huayu Lin 1, Shuqiong Wen 2, Qiao Liu 1, Yiding Li 3, Qing Wu 1, Leiqiong Gao 1, Xiangyu Chen 1, Luoyingzi Xie 1, Qin Tian 1, Jianfang Tang 1, Zhirong Li 1, Li Hu 1, Juan Wang 4, Lifan Xu 1, Qizhao Huang 5, Lilin Ye 1
1Institute of Immunology, Third Military Medical University, 2Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Stomatological Hospital, Sun Yat-Sen University, 3Shigatse Branch, Xinqiao Hospital, Third Military Medical University, 4Department of Emergency Medicine, Southwest Hospital, Third Military Medical University, 5Cancer Center, The General Hospital of Western Theater Command

Here, we present a tumor transplantation protocol for the characterization of tumor-inherent and periphery-derived tumor-infiltrated lymphocytes in a mouse tumor model. Specific tracing of the influx of recipient-derived immune cells with flow cytometry reveals the dynamics of the phenotypic and functional changes of these cells during antitumor immune responses.

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Medicine

In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter
Zhaokai Li *1,2, Liu Liu *1, Zhong Wang 1
1Department of Cardiac Surgery, Cardiovascular Center, The University of Michigan, Ann Arbor, 2Department of Cardiovascular Medicine, Xiangya Hospital, Central South University

We describe here, the establishment and application of an Tg(Myh6-cre)1Jmk/J /Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze/J (referred to as αMHC-Cre/Rosa26A-Flox-Stop-Flox-GCaMP3 below) mouse reporter line for cardiac reprogramming assessment. Neonatal cardiac fibroblasts (NCFs) isolated from the mouse strain are converted into induced cardiomyocytes (iCMs), allowing for convenient and efficient evaluation of reprogramming efficiency and functional maturation of iCMs via calcium (Ca2+) flux.

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Developmental Biology

An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells
Tram P. Le *1, Xiaolei Zhao *1, Shannon Erhardt 1,2, Jianhua Gu 3, Huie Wang 3, Tina O. Findley 1, Jun Wang 1,2
1Department of Pediatrics, McGovern Medical School, The University of Texas Health Science Center at Houston, 2The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston, 3SEM AFM Core in the Houston Methodist Hospital Research Institute

Detailed step-by-step protocols are described here for studying mechanical signals in vitro using multipotent O9-1 neural crest cells and polyacrylamide hydrogels of varying stiffness.

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Medicine

Robot-assisted Total Mesorectal Excision and Lateral Pelvic Lymph Node Dissection for Locally Advanced Middle-low Rectal Cancer
Chenhao Hu *1,2,3, Zhe Zhang *1,2,3, Lei Zhang 1,2,3, Ruihan Liu 2,3, Jun Yan 1,2,3, Qi Sun 1,2,3, Guanghui Wang 1, Junjun She 1,2,3
1Department of General Surgery, The First Affiliated Hospital of Xian Jiaotong University, 2Center for Gut Microbiome Research, Med-X Institute, The First Affiliated Hospital of Xian Jiao tong University, 3Department of High Talent, The First Affiliated Hospital of Xian Jiaotong University

The robotic technique described herein aims to detail a stepwise approach to robot-assisted total mesorectal excision and lateral pelvic lymph node dissection for locally advanced (T3/T4) rectal cancer located below the peritoneal reflection.

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Chemistry

Synthesis of Metal Nanoparticles Supported on Carbon Nanotube with Doped Co and N Atoms and its Catalytic Applications in Hydrogen Production
Pui-Ching Poon *1, Ka-Man Lee *1, Yuanhao Wang 2, William Wai Yan Lam 1, Peter Shu-Wai Leung 1, Xiao-Ying Lu 1, Weiqun Li 3, B. Layla Mehdi 3, Yang Lu 4, Chi-Wing Tsang 1, Eugene Yin Cheung Wong 2
1Faculty of Science and Technology, Technological and Higher Education Institute of Hong Kong (THEi), 2Department of Supply Chain and Information Management, The Hang Seng University of Hong Kong, 3Department of Mechanical, Materials and Aerospace Engineering, University of Liverpool, 4Laboratory of Building Environment and New Energy Resources, School of Civil Engineering, Faculty of Infrastructure Engineering, Dalian University of Technology

Here, we present a protocol to synthesize Co nanoparticles supported on carbon nanotubes with Co- and N- dopants for hydrogen productions.

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Biology

Assessment of Cellular Bioenergetics in Mouse Hematopoietic Stem and Primitive Progenitor Cells using the Extracellular Flux Analyzer
Surinder Kumar 1, Morgan Jones 2, Qing Li 2,3,4, David B. Lombard 1,4,5
1Department of Pathology, University of Michigan, Ann Arbor, 2Department of Internal Medicine, University of Michigan, Ann Arbor, 3Department of Cellular and Developmental Biology, University of Michigan, Ann Arbor, 4Rogel Cancer Center, University of Michigan, Ann Arbor, 5Institute of Gerontology, University of Michigan, Ann Arbor

The method presented here summarizes optimized protocols for assessing cellular bioenergetics in non-adherent mouse hematopoietic stem and primitive progenitor cells (HSPCs) using the extracellular flux analyzer to measure the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) of HSPCs in real time.

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Medicine

Aggravation of Myocardial Ischemia upon Particulate Matter Exposure in Atherosclerosis Animal Model
Yuanmin Yang *1, Shuoqiu Deng *1, Shuiqing Qu 1, Yu Zhang 1, Zhongyuan Zheng 1, Lina Chen 1, Yujie Li 1
1China Academy of Chinese Medical Sciences

This protocol describes a composite animal model with exposure to particulate matter (PM) that aggravates myocardial ischemia with atherosclerosis.

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Medicine

Application of Straight-needle, Three-tailed, Knot-free, Peritoneal Sutures in Laparoscopic Transabdominal Preperitoneal Hernia Repair
Ning Ma *1, Zhen Zong *2, Fu-xing Tang 1, En-min Huang 1, Tao Ma 1, Wei-sheng Yang 1, Jia-lin Chen 1, Shuang Chen 1, Tai-cheng Zhou 1
1Department of Gastroenterological Surgery and Hernia Center, the Sixth Affiliated Hospital of Sun Yat-sen University, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, 2Department of Gastrointestinal Surgery, the Second Affiliated Hospital of Nanchang University

Here, we present a protocol to introduce a new peritoneal suture method. This method is called straight-needle, three-tailed, knot-free suture, and we will outline the manufacturing method and clinical application of this suture in detail.

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Medicine

Laparoscopic Radical Antegrade Modular Pancreatosplenectomy via Dorsal-Caudal Artery Approach for Pancreatic Neck-Body Cancer
Zhantao Shen 1,2, Guihao Chen 1,2, Chunbao Zhu 2, Xiang Wu 1,2, Yifeng Liu 1,2, Zhangyuanzhu Liu 1,2, Zhijian Tan 1,2, Xiaosheng Zhong 1,2
1The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, 2The Second Clinical College of Guangzhou University of Chinese Medicine

With advancements in laparoscopic techniques, laparoscopic radical antegrade modular pancreatosplenectomy (L-RAMPS) has been widely recognized. However, owing to several technical difficulties in this procedure, the artery-first approach in L-RAMPS still remains uncommon. Here, we developed the dorsal-caudal artery approach for L-RAMPS, which might be safe and beneficial for pancreatic neck tumors.

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Medicine

Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis
Li Huang 1, Wei Chen 1, Jiancong Chen 1, Dong Chen 1, Kunsong Zhang 1, Jianpeng Cai 1, Hong Peng 1, Chensong Huang 1, Guangyan Zeng 1, Mingjian Ma 1, Jiahua Liang 1, Borui Xu 1, Xiaoyu Yin 1, Jiaming Lai 1, Lijian Liang 1
1Department of Pancreatobiliary Surgery, The First Affiliated Hospital, Sun Yat-sen University

This protocol presents Indocyanine Green-guided video-assisted retroperitoneal debridement (ICG-guided VARD) for treating severe acute necrotizing pancreatitis.

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Medicine

Laparoscopy-endoscopy Cooperative Surgery for the Treatment of Gastric Gastrointestinal Stromal Tumors
Dongjie Yang *1,2, Xun Hou *1, Wentong Lan *3, Minxuan Shen 1, Shaohua Yang 4, Lele Zhang 1, Qinbo Cai 1, Yan Qian 1, Yuan Lin 5, Xia Feng 6, Yulong He 1,2,4
1Center for Gastrointestinal Surgery, The First Affiliated Hospital, Sun Yat-sen University, 2Guangdong Provincial Key Laboratory of Digestive Cancer Research, 3Department of Endoscopy, The First Affiliated Hospital, Sun Yat-sen University, 4Digestive Medicine Center, The Seventh Affiliated Hospital, Sun Yat-sen University, 5Department of Pathology, The First Affiliated Hospital, Sun Yat-sen University, 6Department of Anesthesiology, The First Affiliated Hospital, Sun Yat-sen University

Laparoscopy-endoscopy cooperative surgery is appropriate for the treatment of gastric gastrointestinal stromal tumors less than 5 cm in size. It can achieve the respective advantages of endoscopic and laparoscopic surgeries while avoiding their drawbacks.

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Biology

Three-Dimensional, Serum-Free Culture System for Lacrimal Gland Stem Cells
Hongzi Chen 1, Pan Huang 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The three-dimensional, serum-free culture method for adult lacrimal gland (LG) stem cells is well established for the induction of LG organoid formation and differentiation into acinar or ductal-like cells.

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Medicine

Microvascular Doppler Assisted Microsurgical Denervation of the Spermatic Cord in Treating Chronic Orchialgia
Jintao Zhuang *1, Mingkuan Zhou *1, Wuyuan Liao 1, Yupeng Gao 1, Xiangan Tu 1
1Department of Urology, The East Division of the First Affiliated Hospital, Sun Yat-sen University

Here, we present a protocol to treat chronic orchialgia by microsurgical denervation of the spermatic cord. Combined with the use of a microvascular doppler, the procedures can be more easily implemented.

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Medicine

Binocular Dynamic Visual Acuity in Eyeglass-Corrected Myopic Patients
Yuexin Wang *1, Yining Guo *1, Shanshan Wei 2, Yifei Yuan 1, Tingyi Wu 1, Yu Zhang 1, Yueguo Chen 1, Xuemin Li 1
1Department of Ophthalmology, Beijing Key Laboratory of Restoration of Damaged Ocular Nerves, Peking University Third Hospital, 2Beijing Tongren Eye Center, Beijing Tongren Hospital

The present research demonstrates a method to accurately examine dynamic visual acuity (DVA) in myopic subjects with eyeglass correction. Further analysis indicated that the closer the refraction state to emmetropia, the better the eyeglass-corrected binocular DVA is at both 40 and 80 degrees per second.

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Developmental Biology

Fixation of Embryonic Mouse Tissue for Cytoneme Analysis
Eric T. Hall 1, Christina A. Daly 1,2, Yan Zhang 1, Miriam E. Dillard 1, Stacey K. Ogden 1
1Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, 2St. Jude Graduate School of Biomedical Sciences

Here, an optimized step-by-step protocol is provided for fixation, immunostaining, and sectioning of embryos to detect specialized signaling filopodia called cytonemes in developing mouse tissues.

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Medicine

The Miniature Pig: A Large Animal Model for Cochlear Implant Research
Xiaojun Ji *1,2,3,4, Yi Luo *1,2,3,4, Weiwei Guo 1,2,3,4, Fei Ji 1,2,3,4, Shuolong Yuan 1,2,3,4, Liangwei Xu 1,2,3,4, Wei Chen *1,2,3,4
1College of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, 2National Clinical Research Center for Otolaryngologic Diseases, 3Key Lab of Hearing Science, Ministry of Education, 4Beijing Key Lab of Hearing Impairment for Prevention and Treatment

Miniature pigs (mini-pigs) are an ideal large animal model for research into cochlear implants. Cochlear implantation surgery in mini-pigs can be utilized to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches in a living system similar to human beings.

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Biology

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
Jinxi Yuan 1, Jie Zhang 1, Yan Zhang 1, WuYun QiQiGe 1, Wei Liu 2, Shanchun Yan 1, Guirong Wang 2
1Key Laboratory of Sustainable Forest Ecosystem Management - Ministry of Education, Northeast Forestry University, 2Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences

This paper presents the step-by-step protocols for CRISPR/Cas9 mutagenesis of the Oriental fruit fly Bactrocera dorsalis. Detailed steps provided by this standardized protocol will serve as a useful guide for generating mutant flies for functional gene studies in B. dorsalis.

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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

Clinical Application of Single-Surgeon, Three-Port, Laparoscopic Resection for Colorectal Cancer with Natural Orifice Specimen Extraction
Xiuqin Cai 1, Qilong Liu 1, Wenfeng Li 1, Junfei He 1, Shibin Yang 1
1Department of Gastrointestinal Surgery, The First Affiliated Hospital, Sun Yat-sen University

Here, we present a protocol to successfully perform single-surgeon, three-port, laparoscopic resection for colorectal cancer with natural orifice specimen extraction.

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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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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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Medicine

Fluorescent Laparoscopic Central Hepatectomy for Liver Cancer Using Indocyanine Green Negative Staining
Jinxing Wei *1, Haixiong Zhang *2, Jinyi Zhong 1, Jiyan Xie 3, Weibang Yang 4, Jinxin Duan 1, Wenfeng Zhuo 1, Ang Li 1, Changzhen Shang 1, Jun Min 1
1Department of Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 2Department of Hepatobiliary Surgery, Foshan Fosun Chancheng Hospital, 3Department of Gastrointestinal Surgery, The Eighth Affiliated Hospital of Sun Yat-sen University, 4Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Guangzhou Medical University

The present protocol describes fluorescent negative staining in laparoscopic central hepatectomy. This technique can make hepatectomy more accurate and precise.

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Medicine

Application of Laparoscopic Hepatectomy Combined with Intraoperative Microwave Ablation in Colorectal Cancer Liver Metastasis
Dawei Zhang *1, Zuxiao Chen *2, Lei Zhang 1, Xiangjun Qian 1, Xiaoming Huang 1, Zheyu Zheng 1, Weidong Pan 1
1Department of Pancreatic Hepatobiliary Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, 2Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Guangzhou Medical University

This protocol describes the laparoscopic resection of colorectal cancer liver metastases combined with ultrasound-guided microwave ablation. This technique can safely, effectively, and accurately treat refractory liver metastases <3 cm, reduce postoperative complications, and accelerate the postoperative rehabilitation of patients.

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Medicine

Application of a New Mesh Fixation Method in Laparoscopic Incisional Hernia Repair
Ning Ma *1, Dai Tang *2, Fu-xing Tang 1, En-min Huang 1, Tao Ma 1, Wei-sheng Yang 1, Chuang-xiong Liu 1, Hao-nan Huang 1, Shuang Chen 1, Tai-cheng Zhou 1
1Department of Gastroenterological Surgery and Hernia Center, The Sixth Affiliated Hospital, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, Guangzhou, Sun Yat-sen University, 2Department of Gastrointestinal Surgery, the Third Affiliated Hospital of Zunyi Medical University (The First People's Hospital of Zunyi)

Presented here is a method for improving the mesh placement in laparoscopic incisional hernia repair, which can shorten the time required for mesh fixation and reduce the occurrence of postoperative chronic pain.

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Fundamental and State-of-the-Art Technologies In Retinal Studies and Disease Repair
Yuan Ma 1, Chunqiao Liu 1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University

Fundamental and State-of-the-Art Technologies In Retinal Studies and Disease Repair

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Medicine

Grade III Varicocele Surgical Treatment using Spermatic Vein-Superficial Abdominal Vein Shunt
Yadong Zhang 1,2, Xiangping Li 3, Deng Chunhua 1
1Department of Andrology, the First Affiliated Hospital, Sun Yat-sen University, 2Health Management Center, the First Affiliated Hospital, Sun Yat-sen University, 3Department of Urology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Southern Medical University

Here, we present a protocol for grade III varicocele surgical treatment that aims to rebuild the venous drainage of the spermatic cord by a spermatic vein-superficial abdominal vein shunt performed under the microscope.

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Neuroscience

A Procedure for Mouse Dorsal Root Ganglion Cryosectioning
Liangliang He *1,2, Wenxing Zhao *1, Lingyi Zhang 2,3, Maalveka Ilango 2,4, Na Zhao 2, Liqiang Yang 1, Zhonghui Guan 2
1Department of Pain Management, Xuanwu Hospital, Capital Medical University, 2Department of Anesthesia and Perioperative Care, University of California San Francisco, 3Department of Anesthesiology, Sun Yat-Sen University, 4University of Washington

Presented here is the development for consistently acquiring high-quality dorsal root ganglion cryostat sections.

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Medicine

Endoscopic Bilateral Nipple-sparing Mastectomy via a Single Axillary Incision with Immediate Pre-pectoral Implant-based Breast Reconstruction
Ze Huang 1, Zongyan Li 1, Xiaofang Zhong 1, Qingyu Yang 1, Lina Wei 1, Dongyuan Li 1, Haiyan Li 1,2
1Division of Breast Surgery, Department of General Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, 2Biomedical Innovation Center, The Sixth Affiliated Hospital, Sun Yat-sen University

Here, we present detailed surgical procedures for endoscopic bilateral nipple-sparing mastectomy via a single axillary incision with immediate pre-pectoral implant-based breast reconstruction. This procedure is a secure and viable method for breast cancer patients.

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Medicine

Combination of High Ligation and Intraoperative Embolization using Polidocanol for Treatment of Varicoceles
Dayu Han 1, Xin Feng 1, Zexin Guo 1, Jintao Zhuang 1, Qiyun Yang 1, Hongcai Cai 1, Chunhua Deng 1, Xiangzhou Sun 1
1Department of Andrology, The First Affiliated Hospital, Sun Yat-sen University

To reduce the postoperative recurrence rate of varicocelectomy, we combined high ligation of varicocele with intraoperative embolization. We injected polidocanol from the spermatic vein during surgery to embolize the branches of the spermatic vein and collateral veins. This is an alternative surgical method for the treatment of varicocele.

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Medicine

Application of Mid-Pancreatectomy with End-to-End Anastomosis in Pancreatic Benign Tumors
Zeyu Lin *1,2, Zejin Lin *1,2, Yongling Liang 1,2, Chengrui Zhong 1,2, Ziyan Mai 1,2, Zhiping Chen 1,2, Jiayan Wu 1,2, Taijun Yi 1,2, Guolin Li 1,2, Yunle Wan 1,2
1Department of General Surgery (Hepatobiliary, Pancreatic and Splenic Surgery), The Sixth Affiliated Hospital, Sun Yat-sen University, 2Biomedical Innovation Center, The Sixth Affiliated Hospital, Sun Yat-sen University

The present protocol describes the application of mid-pancreatectomy combined with end-to-end anastomosis in the surgical treatment of pancreatic benign tumors, which presents a feasible solution for managing such tumors while concurrently preserving pancreatic function.

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Medicine

Iatrogenic Injury Recapitulated: Electroexcision Technique for Urethral Stricture Modeling in Rats
Linlu She 1,2, Bin Niu 1,2, Xiang Wu 1,2, Yinghao Yin 1,2, Kangqiang Wen 1,2, Hu Li 1,2, Dongdong Xie 1,2, Xiaoping Zheng 1,2, Yingbo Dai 1
1Department of Urology, The Fifth Affiliated Hospital of Sun Yat-sen University, 2Guangdong Provincial Key Laboratory of Biomedical Imaging, The Fifth Affiliated Hospital, Sun Yat-Sen University

In this study, we present a novel, efficient, and stable rat model of urethral stricture created through electroexcision of the rat urethra, which effectively simulates iatrogenic injury observed in clinical settings.

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