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Southern Medical University

45 ARTICLES PUBLISHED IN JoVE

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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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Neuroscience

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)
Ying Wang 1, Nicholas Baker 1, Gro V. Amdam 1,2
1School of Life Sciences, Arizona State University , 2Department of Chemistry, Biotechnology, and Food Science, Norwegian University of Life Sciences

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

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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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Immunology and Infection

In Vivo Investigation of Antimicrobial Blue Light Therapy for Multidrug-resistant Acinetobacter baumannii Burn Infections Using Bioluminescence Imaging
Yucheng Wang 1,2,3, Olivia D. Harrington 1, Ying Wang 1, Clinton K. Murray 4, Michael R. Hamblin 1, Tianhong Dai 1
1Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, 2Department of Medical Oncology, Beijing Institute of Translational Medicine, Chinese Academy of Sciences, 3Cancer Center, Aviation General Hospital, Beijing, 4Infectious Disease Service, Brooke Army Medical Center

Infections caused by multidrug-resistant (MDR) bacterial strains have emerged as a serious threat to public health, necessitating the development of alternative therapeutics. We present a protocol to evaluate the effectiveness of antimicrobial blue light (aBL) therapy for MDR Acinetobacter baumannii infections in mouse burns by using bioluminescence imaging.

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Developmental Biology

Use of a Recombinant Mosquito Densovirus As a Gene Delivery Vector for the Functional Analysis of Genes in Mosquito Larvae
Pei-Wen Liu 1, Jia-Bao Xu 1, Yun-Qiao Dong 2, Xiao-Guang Chen 1, Jin-Bao Gu 1
1Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, 2Reproductive Medical Centre of Guangdong Women and Children's Hospital

We report using an artificial intronic small RNA expression strategy to develop a non-defective recombinant Aedes aegypti densovirus (AaeDV) in vivo delivery system. A detailed procedure for the construction, packaging, and quantitative analysis of the rAaeDV vectors as well as for larval infection is described.

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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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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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Neuroscience

Auditory Brainstem Response and Outer Hair Cell Whole-cell Patch Clamp Recording in Postnatal Rats
Aoshuang Chang 1,2, Cuixian Li 1, Jianfeng Huang 1,3, Wenlu Pan 1, Yinghong Tian 4, Jie Tang 1
1Department of Physiology, School of Basic Medical Sciences, Southern Medical University, 2School of Basic Medical Sciences, Guizhou Medical University, 3Department of Laboratory Medicines, Guangzhou General Hospital of Guangzhou Military Region, Southern Medical University, 4Experiment Teaching Center, School of Basic Medical Sciences, Southern Medical University

This protocol describes methods to record the auditory brainstem response from postnatal rat pups. To examine the functional development of outer hair cells, the experimental procedure of whole-cell patch clamp recording in isolated outer hair cells is described step-by-step.

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Cancer Research

A Comprehensive Procedure to Evaluate the In Vitro Performance of the Putative Hemangioblastoma Neovascularization Using the Spheroid Sprouting Assay
Ying Wang 1, DanQi Chen 1, MingYu Chen 1, KaiYuan Ji 1, DeXuan Ma 1, LiangFu Zhou 1
1Department of Neurosurgery, Huashan Hospital, Fudan University

This paper presents a comprehensive procedure to evaluate in vitro whether classic tumor angiogenesis exists in hemangioblastomas (HBs) and its role in HBs. The results highlight the complexity of HB-neovascularization and suggest that this common form of angiogenesis is only a complementary mechanism in the HB-neovascularization.

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Engineering

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7δ/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 (STO) Single-crystal Substrates
Ying Wang 1,2, Zhen Li 1,2, YongSheng Liu 1, Yijie Li 2, Linfei Liu 2, Da Xu 2, Xiaojing Luo 1, Tian Gao 1,3, Yanyan Zhu 1,4, Luozeng Zhou 3, Jianming Xu 3
1Department of Physics, Mathematics, Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, 2Key Laboratory of Artificial Structure and Quantum Control, Ministry of Education, Department of Physics, Shanghai Jiao Tong University, 3Shanghai Institute of Space Power-sources, 4Shanghai Key Laboratory of High Temperature Superconductors, Shanghai University

Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF)-sputtering.

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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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Developmental Biology

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome
Ping Xue 1, Zhiqiang Wang 1, Xiaomin Fu 1,2, Junjiang Wang 1,3, Gopika Punchhi 1, Andrew Wolfe 1,4, Sheng Wu 1,4,5
1Department of Pediatrics, Johns Hopkins University School of Medicine, 2Department of Health, Beijing Military General Hospital, 3Southern Medical University, 4Department of Molecular and Cellular Physiology, Johns Hopkins University School of Medicine, 5Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine

We describe the development of a lean PCOS-like mouse model with dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

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Medicine

Mechanical Micronization of Lipoaspirates for Regenerative Therapy
Huidong Zhu 1, Jinbo Ge 1, Xihang Chen 1, Feng Lu *1, Junrong Cai *1
1Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University

Here, we present a protocol to obtain stromal vascular fraction from adipose tissue through a series of mechanical processes, which include emulsification and multiple centrifugations.

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Immunology and Infection

Human Serum Anti-aquaporin-4 Immunoglobulin G Detection by Cell-based Assay
Caiyun Liu 1, Mingqin Zhu 1, Ying Wang 1
1Department of Neurology and Neuroscience Center, The First Hospital of Jilin University

Cell-based assay is a widely used method to detect serum anti-aquaporin-4 immunoglobulin G. This method could be applied to clinical diagnosis and scientific researches of neuromyelitis optical spectrum disorders.

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Biology

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice
Qiancheng Wang 1, Kaitong Chen 1, Hairuo Lin 1, Mingyuan He 2, Xiaoxia Huang 1, Hailin Zhu 1, Yulin Liao 1
1Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 2The Third Clinical Medical College, Guangzhou Medical University

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

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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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Genetics

A Bioinformatics Pipeline to Accurately and Efficiently Analyze the MicroRNA Transcriptomes in Plants
Ying Wang *1,2, Zheng Kuang *1,2, Lei Li 2, Xiaozeng Yang 1
1Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, 2State Key Laboratory of Protein and Plant Gene Research, Peking-Tsinghua Center for Life Sciences, School of Advanced Agricultural Sciences and School of Life Sciences, Peking University

A bioinformatics pipeline, namely miRDeep-P2 (miRDP2 for short), with updated plant miRNA criteria and an overhauled algorithm, could accurately and efficiently analyze microRNA transcriptomes in plants, especially for species with complex and large genomes.

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Immunology and Infection

Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models
Soichi Sano 1, Ying Wang 1, Megan A. Evans 1, Yoshimitsu Yura 1, Miho Sano 1, Hayato Ogawa 1, Keita Horitani 1, Heather Doviak 1, Kenneth Walsh 1
1Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

Described are protocols for the highly efficient genome editing of murine hematopoietic stem and progenitor cells (HSPC) by the CRISPR/Cas9 system to rapidly develop mouse model systems with hematopoietic system-specific gene modifications.

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Developmental Biology

An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo
Yunyun Jiang 1, Jiahao Chen 1, Jin Xu 2
1Department of Developmental Biology, School of Basic Medical Sciences, Southern Medical University, 2Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology

To test the effect of a chemokine on macrophage recruitment in vivo, the whole mount in situ hybridization was used to detect the ectopic expression of the chemokine, and immunostaining was used to label macrophages. Live imaging was used for real-time observation of macrophage migration.

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Behavior

Quantitative Static and Dynamic Assessment of Balance Control in Stroke Patients
Qiang Lin *1,2,3,4, Yuxin Zheng *1,2,3,4, Peixi Lian 3,4, Yuanyuan Guo 1,3,4, Haojie Huang 5, Zhiwei Luo 1,2, Wanying Li 1,2, Hongxin Chen 1,2,3,4, Sufen Li 1, Shijuan Lang 1,2,4, Yumei Chen 6, Haining Ou *1,2,3,4, Junjie Liang *1,2,3,4
1Department of Rehabilitation, The Fifth Affiliated Hospital of Guangzhou Medical University, 2The Rehabilitation Medicine Lab of Guangzhou Medical University, 3Experiment Education Model Center of Rehabilitation Medicine, Guangzhou Medical University, 4Department of Rehabilitation Therapy, Guangzhou Medical University, 5College of Physical Education and Sports, Beijing Normal University, 6Department of Rehabilitation, Nanfang Hospital, Southern Medical University

Presented here is a quantitative, clinical balance assessment method suitable for stroke patients with balance disorders.

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Bioengineering

Multiphoton Microscopic Observation of Vessels in Mouse Liver Tissue
Wen Rongrong *5, Li Ru *1, He Sixiao 1, Wang Ziqing 1, Huang Junhao 1, Zhao Liying 2, Tian Zhihui 3,4, Ma Qiang 1
1Department of Biopharmaceutics, School of Laboratory Medicine and Biotechnology, Southern Medical University, 2Department of General Surgery, Nanfang Hospital, Southern Medical University, 3Department of Stomatology, Nanfang Hospital, Southern Medical University, 4College of Stomatology, Southern Medical University, 5School of Laboratory Medicine and Biotechnology, Southern Medical University

In this experiment, a mouse is injected in its tail vein with Rhodamine B isothiocyanate–dextran that can stain blood vessels. After the liver is exposed and fixed, a specific part of the liver can be selected to observe the deep tissue in the living body using multiphoton microscopy.

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Medicine

Evaluation of Color Difference in Placenta with Twin Anemia Polycythemia Sequence
Depeng Zhao 1, Lisanne S.A. Tollenaar 2, Femke Slaghekke 2, Dick Oepkes 2, Tao Duan 3, Enrico Lopriore 4
1Department of Reproductive Medicine, Affiliated Shenzhen Maternity and Child Healthcare Hospital, Southern Medical University, 2Division of Fetal Medicine, Department of Obstetrics, Leiden University Medical Center, 3Fetal Medicine Unit & Prenatal Diagnosis Center, Department of Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, 4Division of Neonatology, Department of Pediatrics, Leiden University Medical Center

In monochorionic twins with twin anemia polycythemia sequence (TAPS), the donor twin and its corresponding placenta share are pale, while the recipient twin and its placental share have a plethoric aspect. Presented here is a protocol to quantify the color difference in maternal side of TAPS placenta after birth.

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Medicine

Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy
Yi Lao 1, Cankun Zheng 1, Hailin Zhu 1,2, Hairuo Lin 1, Xiaoxia Huang 1, Yulin Liao 1
1Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 2Department of Geriatrics, Pingxiang People's Hospital of Southern Medical University

This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning.

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JoVE Core

Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis
Eunbee Park 1, Megan A. Evans 4, Heather Doviak 4, Keita Horitani 4, Hayato Ogawa 4, Yoshimitsu Yura 4, Ying Wang 2, Soichi Sano 3,4, Kenneth Walsh 1,4
1Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, 2Department of Cardiology, Xinqiao Hospital, Army Medical University, 3Department of Cardiology, Osaka City University Graduate School of Medicine, 4Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

We describe three methods of bone marrow transplantation (BMT): BMT with total-body irradiation, BMT with shielded irradiation, and BMT method with no pre-conditioning (adoptive BMT) for the study of clonal hematopoiesis in mouse models.

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Genetics

A Virtual Machine Platform for Non-Computer Professionals for Using Deep Learning to Classify Biological Sequences of Metagenomic Data
Zhencheng Fang 1,2, Hongwei Zhou 1,3
1Microbiome Medicine Center, Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, 2Center for Quantitative Biology, Peking University, 3State Key Laboratory of Organ Failure Research, Southern Medical University

This tutorial describes a simple method to construct a deep learning algorithm for performing 2-class sequence classification of metagenomic data.

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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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Genetics

Pre-Implantation Genetic Testing for Aneuploidy on a Semiconductor Based Next-Generation Sequencing Platform
Chengming Xu *1, Riqing Wei *2, Hui Lin 1, Leiyu Deng 1, Li Wang 3, Deyang Li 4, Honghui Den 5, Wensong Qin 1, Ping Wen 1, Ying Liu 1, Yingsong Wu 2, Qiang Ma 2, Jinliang Duan 1
1Centre for Women, Children, and Reproduction, Guangxi Key Laboratory of Metabolic Diseases Research, 2Department of Biopharmaceutics, School of Laboratory Medicine and Biotechnology, Southern Medical University, 3Department of Obstetrics and Gynecology, Chinese PLA General Hospital, 4Clinical Laboratory, Northern Theater Air Force Hospital, 5Guangzhou Darui Reproduction Technology Co., Ltd.

The protocol presents the overall in-lab procedures required in pre-implantation genetic testing for aneuploidy on a semiconductor-based next-generation sequencing platform. Here we present the detailed steps of whole genome amplification, DNA fragment selection, library construction, template preparation, and sequencing working flow with representative results.

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Medicine

Generation and Characterization of Right Ventricular Myocardial Infarction Induced by Permanent Ligation of the Right Coronary Artery in Mice
Ruoxi Liao 1, Mingyuan He 2, Donghong Hu 2, Chengzheng Gong 1, Huiying Du 1, Hairuo Lin 2, Huijun Sun 3
1The First Clinical College of Dalian Medical University, 2Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, 3College of Pharmacy, Dalian Medical University

There are several differences between the right and left ventricles. However, the pathophysiology of right ventricular infarction (RVI) has not been clarified. In the present protocol, a reproducible method for RVI mouse model generation is introduced, which may provide a means to explain the mechanism of RVI.

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Bioengineering

Fabricating Highly Open Porous Microspheres (HOPMs) via Microfluidic Technology
Sheng-Chang Luo 1,2, Ying Wang 3, Ranjith Kumar Kankala 1,2, Yu Shrike Zhang 4, Ai-Zheng Chen 1,2
1Institute of Biomaterials and Tissue Engineering, Huaqiao University, 2Fujian Provincial Key Laboratory of Biochemical Technology, Huaqiao University, 3Affiliated Dongguan Hospital, Southern Medical University, 4Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School

The present protocol describes the fabrication of poly(lactic-co-glycolic acid)-based highly open porous microspheres (HOPMs) via the single-emulsion formulation based facile microfluidic technology. These microspheres have potential applications in tissue engineering and drug screening.

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Neuroscience

Using a Bipolar Electrode to Create a Temporal Lobe Epilepsy Mouse Model by Electrical Kindling of the Amygdala
Yongchang Lu *1,2, Yang Dai *1,2, Siqi Ou 1,2, Yujing Miao 1, Ying Wang 1,2, Quanlei Liu 1,2, Yihe Wang 1,2, Penghu Wei 1,2, Yongzhi Shan 1,2, Guoguang Zhao 1,2,3
1Department of Neurosurgery, Xuanwu Hospital Capital Medical University, 2China Clinical Research Center for Epilepsy Capital Medical University, 3China Beijing Municipal Geriatric Medical Research Center

The amygdala plays a key role in temporal lobe epilepsy, which originates in and propagates from this structure. This article provides a detailed description of the fabrication of deep brain electrodes with both recording and stimulating functions. It introduces a model of medial temporal lobe epilepsy originating from the amygdala.

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Medicine

Bloodless Laparoscopic Partial Splenectomy Assisted by Bipolar Radiofrequency Excision Hemostatic Device
Lei Cai *1, Cheng Zhang *1, Kai-hang Zhong 1, Guo-lin He 1, Yuan Cheng 1, Shun-jun Fu 1, Ming-xin Pan 1
1Department of Hepatobiliary Surgery II, General Surgery Center, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University

The present protocol describes that bipolar radiofrequency device-assisted laparoscopic partial splenectomy (LPS) is safe and effective. A bipolar radiofrequency device in LPS can reduce intraoperative bleeding and achieve the clinical effect of a "bloodless spleen incision", which is worthy of clinical application.

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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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Medicine

Plasma Polishing as a New Polishing Option to Reduce the Surface Roughness of Porous Titanium Alloy for 3D Printing
Zhiwei Lin *1,2, Lincong Luo *3, Dongxin Lin *4, Yuping Deng 4,5, Yang Yang 4, Xuecheng Huang 6, Tingrui Wu 1, Wenhua Huang 1,2,4
1Orthopedic Center, Affiliated Hospital of Guangdong Medical University, 2The School of Basic Medical Sciences, Guangdong Medical University, 3The School of Basic Medical Sciences, Fujian Medical University, 4Guangdong Engineering Research Center for Translation of Medical 3D Printing Application, Guangdong Provincial Key Laboratory of Digital Medicine and Biomechanics, National Key Discipline of Human Anatomy, School of Basic Medical Sciences, Southern Medical University, 5Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, 6Shenzhen Hospital of Guangzhou University of Chinese Medicine (Futian)

Plasma polishing is a promising surface processing technology, especially suitable for 3D printing of porous titanium alloy workpieces. It can remove semi-molten powders and ablative oxide layers, thereby effectively reducing surface roughness and improving surface quality.

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Medicine

Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue
Jinru Song 1, Yue Zhang 1, Huihui Pan 1, Xueyan Xu 1, Cheng-Cheng Deng 1, Bin Yang 1
1Dermatology Hospital, Southern Medical University

This study describes an optimized protocol for establishing primary fibroblasts from keloid tissues that can effectively and steadily provide pure and viable fibroblasts.

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Behavior

Laparoscopic Anterior Right Hepatectomy: A Single-Center Experience
Cheng Zhang 1, Chang Li 1, Chunming Wang 1, Lei Cai 1, Guolin He 1, Shunjun Fu 1, Mingxin Pan 1
1Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University

Here we present a step-by-step protocol for conducting laparoscopic anterior right hepatectomy and compare its clinical effects and postoperative outcomes with those of conventional hepatectomies. Analysis of the data of 82 patients with hepatocellular carcinoma revealed that laparoscopic anterior right hepatectomy had better clinical outcomes and survival rates than the conventional hepatectomy.

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Biology

Isolation and Purification of Bacterial Extracellular Vesicles from Human Feces Using Density Gradient Centrifugation
Yicong Xue *1, Xixin Huang *1, Zihao Ou *1, Yuanyuan Wu 1, Qianbei Li 1, Xinyue Huang 1, Minghui Wen 1, Yan Yang 1, Bo Situ 1, Lei Zheng 1
1Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University

This study describes a method to isolate and purify bacterial extracellular vesicles (BEVs) enriched from human feces via density gradient centrifugation (DGC), identifies the physical characteristics of BEVs from morphology, particle size, and concentration, and discusses the potential applications of the DGC approach in clinical and scientific research.

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Immunology and Infection

Quantitative Polymerase Chain Reaction (qPCR)-Based Rapid Diagnosis of Helicobacter pylori Infection and Antibiotic Resistance
Liang Wang *1,2,3, Jin-Xin Lai *1, Yu-Ting Si *1,4, Xu-Xia Cui 1,4, Zeeshan Umar 1,5, Xiao-Jun Ru 1, Xin-Yu Zhang 1, Zheng-Kang Li 1, Alfred Chin Yen Tay 5,6,7,8, Barry J. Marshall 5,6,7,8, Guang-Hua Li 1, Bing Gu 1
1Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, 2Division of Microbiology and Immunology, School of Biomedical Sciences, The University of Western Australia, 3Center for Precision Health, School of Medical and Health Sciences, Edith Cowan University, 4Medical Technology School of Xuzhou Medical University, 5Marshall Laboratory of Biomedical Engineering, School of Medicine, Shenzhen University, 6The Marshall Centre for Infectious Diseases Research and Training, The University of Western Australia, 7Marshall International Digestive Diseases Hospital, Zhengzhou University, 8Marshall Medical Research Center, Fifth Affiliated Hospital of Zhengzhou University

The protocol presents a noninvasive method for the rapid diagnosis of Helicobacter pylori stomach infections through the string test and determines its antibiotic resistance to clarithromycin and levofloxacin using quantitative polymerase chain reaction (qPCR).

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Medicine

Duodenal-Jejunal Bypass Surgery in Diet-Induced Obese Diabetic Mice
Jipei He *1,2, Heng Li *1, Xiaojiang Dai 1, Zhengyong Xie 2, Zhigao Song 3, Xi Chen 2, Hongyan Huang 1, Yunfa Ding 1, Tengfei Qi 1, Qing Liu 4, Hongbin Zhang 2,5, Liangping Wu 1,6
1Jinshazhou Hospital of Guangzhou University of Chinese Medicine, 2General Hospital of Southern Theatre Command, 3Department of Cardiovascular Surgery, Zhujiang Hospital of Southern Medical University, 4Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 5School of Laboratory and Biotechnology, Southern Medical University, 6Guangzhou Hualiang Qingying Biotechnology Co. Ltd

Duodenal-jejunal bypass (DJB) surgery can improve glucose metabolism and reduce insulin resistance. Here, we present a protocol to establish a stable and reliable mouse model of DJB.

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Medicine

Multi-Gene Single Nucleotide Polymorphism Detection in Gastric Cancer Based on Ion Semiconductor Sequencing Platform
Kaihua Huang 1, Xiao Yan 1, Zhicheng Huang 1, Huayuan Liang 1, Zhiwei Li 1, Minghao Wang 1, Yong Wan 2, Suihai Wang 3,4, Liying Zhao 1
1Nanfang Hospital, Southern Medical University, 2Research and Development Department, Guangzhou Darui Biotechnology Co., Ltd, 3Key Laboratory of Antibody Engineering of Guangdong Higher Education Institutes, School of Laboratory Medicine and Biotechnology, Southern Medical University, 4Provincial Key Laboratory of Immune Regulation and Immunotherapy, Southern Medical University

This protocol proposes holistic laboratory procedures required to detect single nucleotide polymorphisms in gastric cancer samples based on an ion semiconductor sequencing platform. The target sequences, ligating adapters, library amplification and purification, and quality control criteria are also described in detail.

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Immunology and Infection

Rapid Detection of Bacterial Pathogens Causing Lower Respiratory Tract Infections via Microfluidic-Chip-Based Loop-Mediated Isothermal Amplification
Jin-Xin Lai *1,2, Yu-Rong Qin *1,3, Yi-Wen Liao *4, Yu-Ting Si 1, Quan Yuan 1, Shi-Mei Huang 2, Yu-Rong Tang 2, Ji-Liang Wang 2, Liang Wang 1
1Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, 2Department of Laboratory Medicine, Shengli Oilfield Central Hospital, 3Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, 4Dermatology Hospital, Southern Medical University

Various bacterial pathogens can cause respiratory tract infections and lead to serious health issues if not detected accurately and treated promptly. Rapid and accurate detection of these pathogens via loop-mediated isothermal amplification provides effective management and control of respiratory tract infections in clinical settings.

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Immunology and Infection

Accessing Early Differentiation of Virus-Specific Follicular Helper CD4+ T Cell in Acute LCMV-Infected Mice
Yao Lin *1,2, Shuai Yue *3, Yang Yang *4, Junjian He 5, Xiaofan Yang 6, Lilin Ye 5, Xiangyu Chen 7
1Department of Urology, South China Hospital, Medical School, Shenzhen University, 2Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Shenzhen University Medical School, 3Cancer Center, Daping Hospital & Army Medical Center of PLA, Third Military Medical University, 4Guangdong Provincial Key Laboratory of Immune Regulation and Immunotherapy, School of Laboratory Medicine and Biotechnology, Southern Medical University, 5Institute of Immunology, Third Military Medical University, 6Dermatology Hospital, Southern Medical University, 7Institute of Immunological Innovation and Translation, Chongqing Medical University

The current study showcases protocols for assessing the early fate commitment of virus-specific TFH cells and manipulating gene expression in these cells.

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Biology

Co-culture Model Using Two Types of Adherent Cell Lines
Zhuo Song *1, Lisha Zhao *1, Jingwen Hu *1, Xi Zheng 2, Yingyu Liu 1, Hao Wang 1, Xiaoyan Chen 1
1Maternal-Fetal Medicine Institute, Department of Obstetrics and Gynecology, Shenzhen Baoan Women's and Children's Hospital, Shenzhen University, 2The First Clinical Medicine College, Southern Medical University

The protocol shows a novel in vitro experimental model that can recapitulate the biology of two kinds of adherent cell lines with a three-dimensional (3D)-printed scaffold. The construction of this model and operating procedures, from cell preparation and cell culture to analysis and evaluation, are described.

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Bioengineering

A Medical Decision Support Platform for Identifying Thrombospondin 1 in Non-alcoholic Fatty Liver Disease via Immuno-Infiltration Analysis
Jianyu Huang 1,2,3, Pengfei Li 1,4, Jianing Chi 1,4, Jiaman Hu 1, Hua Cai 1, Ningxia Wu 1, Min Li 1, Zhongqiu Lin 1, Yutong Ji 1, Jiajia Xu 1, Peng Liu 5, Senthil Kumar Jagatheesaperumal 6, Victor Hugo C. de Albuquerque 7, Lin Xu 1,2,3,4
1Department of Geriatric Cardiology, General Hospital of Southern Theater Command, 2Guangzhou Key Laboratory of Cardiac Rehabilitation, 3Branch of National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital, 4The First School of Clinical Medicine, Southern Medical University, 5Zhujiang Hospital, Southern Medical University, Cardiovascular Department/The Second School of Clinical Medicine, Southern Medical University, 6Department of Electronics & Communication Engineering, Mepco Schlenk Engineering College, 7Department of Teleinformatics Engineering, Federal University of Ceará

This study investigated the relationships between non-alcoholic fatty liver disease (NAFLD) and myocardial infarction (MI) through co-expressed genes, identifying Thrombospondin 1 (THBS1) as a biomarker. Immuno-infiltration analysis revealed CD8+ T cells and neutrophils as key factors, with THBS1 showing potential as a diagnostic tool for NAFLD and MI.

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