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32 ARTICLES PUBLISHED IN JoVE

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Biology

Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region
Kin Chiu 1, Wui Man Lau 1, Ho Tak Lau 1, Kwok-Fai So 1, Raymond Chuen-Chung Chang 1
1Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong - HKU

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

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Biology

Intravitreous Injection for Establishing Ocular Diseases Model
Kin Chiu 1, Raymond Chuen-Chung Chang 1, Kwok-Fai So 1
1The University of Hong Kong - HKU

Intravitreous injection is a widely used technique in visual sciences research for ocular diseases or as direct application of local treatment. This video demonstrated a protocol for intravitreous injection using a 1ml syringe with glass pipette. Useful tips about avoiding massive bleeding and lens damage are given.

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Biology

Laser-Induced Chronic Ocular Hypertension Model on SD Rats
Kin Chiu 1, Raymond Chang 1, Kwok-Fai So 1
1Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong - HKU

Glaucoma is one of the major causes of blindness in the world. Elevated intraocular pressure is a major risk factor. Laser photocoagulation induced ocular hypertension is one of the well established animal models. This video demonstrates how to induce ocular hypertension by Argon laser photocoagulation in rat.

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Biology

Retrograde Labeling of Retinal Ganglion Cells by Application of Fluoro-Gold on the Surface of Superior Colliculus
Kin Chiu 1, Wui-Man Lau 1, Sze-chun Yeung 1, Raymond Chuen-Chung Chang 1, Kwok-Fai So 1
1The University of Hong Kong - HKU

This video describes the method of retrograde labeling of RGC by applying fluoro-gold (FG) on the surface of superior colliculus (SC). Technique involves drilling the skull, aspirating the cortex, and applying gelatin sponge over entire dorsal surface of SC.

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Neuroscience

Morphometric Analyses of Retinal Sections
Tin Fung Chan 1, Kin Chiu 1, Carmen Ka Ming Lok 1, Wing Lau Ho 1, Kwok-Fai So 1,2,3, Raymond Chuen-Chung Chang 1,2,3
1Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong, 2Research Centre of Heart, Brain, Hormone and Healthy Aging, LKS Faculty of Medicine, The University of Hong Kong, 3State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong

This video demonstrates three types of morphometric analyses of the retina, which include measuring the inner nuclear layer thickness, quantifying the number of retinal ganglion cells (RGCs) and measuring the sizes of RGCs. The technique can offer a simple but scientific platform for morphometric analyses.

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Chemistry

Preparation and Characterization of SDF-1α-Chitosan-Dextran Sulfate Nanoparticles
Andrew R. Bader 1, Tina Li 1, Weiping Wang 2, Daniel S. Kohane 2, Joseph Loscalzo 1, Ying-Yi Zhang 1
1Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, 2Laboratory for Biomaterials and Drug Delivery, Department of Anesthesiology, Division of Critical Care, Boston Children's Hospital

The objective of this protocol is to incorporate SDF-1α, a stem cell homing factor, into dextran sulfate-chitosan nanoparticles. The resultant particles are measured for their size and zeta potential, as well as the content, activity, and in vitro release rate of SDF-1α from the nanoparticles.

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Biology

Characterization of Calcification Events Using Live Optical and Electron Microscopy Techniques in a Marine Tubeworm
Vera B. S. Chan 1, Takashi Toyofuku 2, George Wetzel 3, Laxmikant Saraf 3, Vengatesen Thiyagarajan 4, Andrew S. Mount 1
1Department of Biological Sciences, Clemson University, 2Department of Marine Biodiversity Research (BioDive), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 3Advanced Material Research Laboratory (AMRL), Clemson University, 4Swire Institute of Marine Sciences and School of Biological Sciences, The University of Hong Kong

We demonstrate the use of various microscopy methods that are useful in observing the calcification of a tubeworm, Hydroides elegans, as well as locating and characterizing the first calcified material. Live microscopy and electron microscopy are used together to provide functional and material information that are important in studying biomineralization.

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Medicine

Minimally Invasive Transverse Aortic Constriction in Mice
Aung Moe Zaw 1, Connor M. Williams 2, Helen K. W. Law 3, Billy Kwok Chong Chow 1
1School of Biological Sciences, The University of Hong Kong, 2College of Arts and Science, New York University, 3Department of Health Technology and Informatics, Faculty of Health and Social Sciences, The Hong Kong Polytechnic University

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

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JoVE Core

Using Wavelet Entropy to Demonstrate how Mindfulness Practice Increases Coordination between Irregular Cerebral and Cardiac Activities
Hin Hung Sik 1, Junling Gao 1,2, Jicong Fan 1, Bonnie Wai Yan Wu 1, Hang Kin Leung 1, Yeung Sam Hung 2
1Centre of Buddhist Studies, The University of Hong Kong, 2Department of Electrical and Electronic Engineering, The University of Hong Kong

This manuscript describes how to use the wavelet entropy index to analyze high-density electroencephalography (EEG) and electrocardiography (ECG) data. We show that the irregularity of cerebral and cardiac activities became more coordinated during mindfulness-based stress reduction practice.

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Developmental Biology

Hypoxic Preconditioning of Marrow-derived Progenitor Cells As a Source for the Generation of Mature Schwann Cells
Yat-Ping Tsui *1, Alan Kwan-Long Mung *1, Ying-Shing Chan 1, Daisy Kwok-Yan Shum 1, Graham Ka-Hon Shea 2
1School of Biomedical Sciences, The University of Hong Kong, 2Department of Orthopaedics and Traumatology, The University of Hong Kong

Marrow stromal cells (MSCs) with neural potential exist within the bone marrow. Our protocol enriches this population of cells via hypoxic preconditioning and thereafter directs them to become mature Schwann cells.

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JoVE Core

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)
Anson H. L. Tang 1, Queenie T. K. Lai 1, Bob M. F. Chung 2, Kelvin C. M. Lee 1, Aaron T. Y. Mok 1, G. K. Yip 1, Anderson H. C. Shum 2, Kenneth K. Y. Wong 1, Kevin K. Tsia 1
1Department of Electrical and Electronic Engineering, The University of Hong Kong, 2Department of Mechanical Engineering, The University of Hong Kong

This protocol describes the implementation of an asymmetric-detection time-stretch optical microscopy system for single-cell imaging in ultrafast microfluidic flow and its applications in imaging flow cytometry.

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Developmental Biology

Zika Virus Infection of Cultured Human Fetal Brain Neural Stem Cells for Immunocytochemical Analysis
Erica L. McGrath 1,2, Junling Gao 1, Ping Wu 1,3
1Department of Neuroscience and Cell Biology, University of Texas Medical Branch, 2Department of Neurology, Johns Hopkins University, 3Beijing Institute for Brain Disorders, Capital Medical University

This article details the methods that are used to expand human fetal brain neural stem cells in culture, as well as how to differentiate them into various neuronal subtypes and astrocytes, with an emphasis on the use of neural stem cells to study Zika virus infection.

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Neuroscience

Screening of Axonal Degeneration in Carpal Tunnel Syndrome Using Ultrasonography and Nerve Conduction Studies
Xue Deng 1, Lai-Heung Phoebe Chau 2, Suk-Yee Chiu 2, Kwok-Pui Leung 3, Yong Hu 1, Wing-Yuk Ip 1
1Department of Orthopedics & Traumatology, The University of Hong Kong, 2Clinical Electro-diagnostic Unit, Tung Wah Hospital, 3Department of Medicine, The University of Hong Kong

Here we present a protocol using nerve conduction studies and ultrasound to screen potential axonal degeneration associated with carpal tunnel syndrome. The criteria for differentiation are established. Compared to conventional approaches, this method is noninvasive, convenient, and efficient, with an overall satisfactory accuracy, sensitivity, and specificity.

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Behavior

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice
Ran You 1,2,3, Yan Liu 3, Raymond Chuen-Chung Chang 3
1Nanjing Key Laboratory of Pediatrics, Children's Hospital of Nanjing Medical University, 2Nanjing Key Laboratory of Pediatrics, Nanjing Medical University, 3Laboratory of Neurodegenerative Disease, Li Ka Shing Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region (HK SAR)

A comprehensive behavioral test battery of motor skills, mood—including social interaction, depression, and anxiety—and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

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Neuroscience

An In Vitro Model for Studying Tau Aggregation Using Lentiviral-mediated Transduction of Human Neurons
Brent Aulston 1, Qing Liu 1, Patrick Reilly 1, Shauna H. Yuan 1
1Department of Neurosciences, University of California, San Diego

This protocol details a procedure in which human neuronal cultures are transduced with lentiviral constructs coding for mutant human tau. Transduced cultures display tau aggregates and associated pathologies.

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Developmental Biology

Hemogenic Endothelium Differentiation from Human Pluripotent Stem Cells in A Feeder- and Xeno-free Defined Condition
Ryo Ohta *1, Ryohichi Sugimura *1, Akira Niwa 1, Megumu K. Saito 1
1Center for iPS cell research and application (CiRA)

Here, we present a protocol to precisely form hemogenic endothelium from human pluripotent stem cells in a feeder- and xeno-free condition. This method will have broad applications in disease modeling and screening for therapeutic compounds.

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Immunology and Infection

Measurement of Chitinase Activity in Biological Samples
Alyssa K. Amick 1, Qing Liu 1, Samir Gautam 1, Geoffrey Chupp 1, Charles S. Dela Cruz *1, Lokesh Sharma *1
1Pulmonary, Critical Care and Sleep Medicine, Yale University School of Medicine

Presented here is a simple method to measure chitinase activity in biological fluids such as bronchoalveolar lavage or serum.

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Behavior

Evaluation of the Cognitive Performance of Hypertensive Patients with Silent Cerebrovascular Lesions
Manman Zhang 1,2, Junling Gao 3, Bingjiao Xie 2, Henry Ka Fung Mak 4, Raymond Tak Fai Cheung 2,5
1Department of Neurology, The First Affiliated Hospital of Wenzhou Medical University, 2Department of Medicine, The University of Hong Kong, 3Centre of Buddhist Studies, The University of Hong Kong, 4Department of Diagnostic Radiology, The University of Hong Kong, 5Research Centre of Heart, Brain, Hormone & Healthy Aging, The University of Hong Kong

Here, we present a protocol to assess whether various types of silent cerebrovascular lesions are differentially associated with deficits in certain cognitive domains in a cohort of 398 hypertensive elderly Chinese, using a combination of neuropsychological tests and multi-sequence 3T MRI scanning.

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Medicine

Intratracheal Administration of Dry Powder Formulation in Mice
Yingshan Qiu 1, Qiuying Liao 1, Michael Y.T. Chow 1,2, Jenny K.W. Lam 1
1Department of Pharmacology and Pharmacy, LKS Faculty of Medicine, The University of Hong Kong, 2Advanced Drug Delivery Group, Sydney Pharmacy School, Faculty of Medicine and Health, The University of Sydney

Dry powder formulations for inhalation have great potential in treating respiratory diseases. Before entering human studies, it is necessary to evaluate the efficacy of the dry powder formulation in preclinical studies. A simple and noninvasive method of the administration of dry powder in mice through the intratracheal route is presented.

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Cancer Research

Tumor Allotransplantation in Drosophila melanogaster with a Programmable Auto-Nanoliter Injector
Shangyu Gong *1, Yichi Zhang *1, Hongcun Bao 1, Xianfeng Wang 1, Chih-Hsuan Chang 1, Yi-Chun Huang 1, Wu-Min Deng 1
1Department of Biochemistry and Molecular Biology, Tulane University School of Medicine

This protocol provides detailed guidance for the initial and continued generational allotransplantation of Drosophila tumors into the abdomen of adult hosts for studying various aspects of neoplasia. Using an autoinjector apparatus, researchers can achieve improved efficiency and tumor yields compared to those achieved by traditional, manual methods.

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Biology

Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [18F]FDG Micro-PET/MR Imaging
Qing Liu *1,2, Kel Vin Tan *3, Hing-Chiu Chang 3, Pek-Lan Khong 3, Xiaoyan Hui 1,2,4
1State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, 2Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 3Department of Diagnostic Radiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 4School of Biomedical Sciences, Institute of Vascular Medicine, Chinese University of Hong Kong

Functional imaging and quantitation of thermogenic adipose depots in mice using a micro-PET/MR imaging-based approach.

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Neuroscience

Modulation of the Neurophysiological Response to Fearful and Stressful Stimuli Through Repetitive Religious Chanting
Hin Hung Sik 1, Georgios T. Halkias 1, Chunqi Chang 2, Junling Gao 1, Hang Kin Leung 1, Bonnie Wai Yan Wu 1
1Buddhism and Science Research Lab, Centre of Buddhist Studies, The University of Hong Kong, 2School of Biomedical Engineering, Shenzhen University

The present event-related potential (ERP) study provides a unique protocol for investigating how religious chanting can modulate negative emotions. The results demonstrate that the late positive potential (LPP) is a robust neurophysiological response to negative emotional stimuli and can be effectively modulated by repetitive religious chanting.

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Biology

Establishing Human Lung Organoids and Proximal Differentiation to Generate Mature Airway Organoids
Cun Li 1,2, Man Chun Chiu 1,2, Yifei Yu 1,2, Xiaojuan Liu 1,2, Ding Xiao 1,2, Jingjing Huang 1,2, Zhixin Wan 1,2, Jie Zhou 1,2,3
1Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 2Centre for Virology, Vaccinology and Therapeutics, Hong Kong Science and Technology Park, 3State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong

The protocol presents a method to derive human lung organoids from primary lung tissues, expand the lung organoids and induce proximal differentiation to generate 3D and 2D airway organoids that faithfully phenocopy the human airway epithelium.

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Medicine

Establishment and Evaluation of a Porcine Vein Graft Disease Model
Xiaohui Li *1, Jinlin Hu *1, Weijiang Tan *2, Zhaoming Lin *1, Caiyi Zhu 2, Chengfeng Huang 1, Jiawen Huang 3, Yintian Liu 1, Qiuying Liao 1, Hua Lu 1, Xiaoshen Zhang 1
1Department of Cardiovascular Surgery, the First Affiliated Hospital of Jinan University, 2Guangdong Provincial Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, 3The First Affiliated Hospital of Jinan University

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.

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Cancer Research

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma
Kel Vin Tan *1,2, Xinxiang Yang *3, Chung Ying Chan 2, Jingjing Shi 1, Hing-Chiu Chang 4, Keith Wan-Hang Chiu 1,5, Kwan Man 3
1Department of Diagnostic Radiology, School of Clinical Medicine, LKS Faculty of Medicine, The University of Hong Kong, 2Department of Oncology, MRC Oxford Institute for Radiation Oncology, University of Oxford, 3Department of Surgery, School of Clinical Medicine, HKU-SZH & LKS Faculty of Medicine, The University of Hong Kong, 4Department of Biomedical Engineering, The Chinese University of Hong Kong, 5Department of Diagnostic and Interventional Radiology, Kwong Wah Hospital

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

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Medicine

Laparoscopic Non-Mesh Cerclage Pectopexy for Pelvic Organ Prolapse
Wenju Zhang *1, Willy Cecilia Cheon *2, Hextan Yuen Sheung Ngan *3, Yuzhen Wei 1, Chaoxia Lyu 1
1Department of Gynecology and Obstetrics, The University of Hong Kong-Shenzhen Hospital, 2Department of Gynecology and Obstetrics, Queen Elizabeth Hospital, 3Department of Obstetrics and Gynecology, Queen Mary Hospital, The University of Hong Kong

The present pilot study describes the development of laparoscopic non-mesh cerclage pectopexy to treat pelvic organ prolapse. The procedure can be used to prevent any complications associated with the use of mesh.

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Developmental Biology

Generation of Natural Killer Cells from Human Expanded Potential Stem Cells
Poon Yu Ching 1, Clement Wang 2, Su Hang 2, Pentao Liu 2,3, Ryohichi Sugimura 2,3
1Major in Molecular Biology and Biotechnology, School of Biological Sciences, The University of Hong Kong, 2School of Biomedical Sciences, The University of Hong Kong, 3Centre for Translational Stem Cell Biology

The present protocol shows how to differentiate CD3−/CD45+CD56+ cells with mild cytotoxicity from human expanded potential stem cells (hEPSCs) under both 3D and 2D culture conditions. This allows for routine phenotypical validation without the destruction of the complex microenvironment.

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Medicine

Facile Preparation and Photoactivation of Prodrug-Dye Nanoassemblies
Yichi Zhang 1,2,3, Kaiqi Long 1,2,3, Weiping Wang 1,2,3
1State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, 2Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 3Laboratory of Molecular Engineering and Nanomedicine, Dr. Li Dak-Sum Research Centre, The University of Hong Kong

This protocol describes the fabrication and characterization of a photoresponsive prodrug-dye nanoassembly. The methodology for drug release from the nanoparticles by light-triggered disassembly, including the light irradiation setup, is explicitly described. The drugs released from the nanoparticles following light irradiation exhibited excellent anti-proliferation effects on human colorectal tumor cells.

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Medicine

Ultrasonographic Evaluation of Salivary Glands for Sjogren's Syndrome: Diagnostic and Monitoring Insights
Iris Y. K. Tang 1, Carmen T. K. Ho 1, Teresa W. L. Li 1, Philip H. Li 1
1Division of Rheumatology and Clinical Immunology, Department of Medicine, Queen Mary Hospital, The University of Hong Kong

Salivary gland ultrasonography is a promising tool for the diagnosis and evaluation of disease activity and complications in Sjogren's syndrome (SS). The scanning technique for salivary gland ultrasound examination relevant to SS is described in the article.

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Neuroscience

Religious Chanting and Self-Related Brain Regions: A Multi-Modal Neuroimaging Study
Hin Hung Sik 1, Stavros Skouras 2, Junling Gao 1, Hang Kin Leung 1, Siu Man Ng 3, Kin Cheung Lee 1, Bonnie Wai Yan Wu 1
1Buddhist Practices and Counselling Science Lab, Centre of Buddhist Studies, The University of Hong Kong, 2Department of Biological and Medical Psychology, Faculty of Psychology, University of Bergen, 3Department of Social Work and Social Administration, The University of Hong Kong, Hong Kong

Here, we present a protocol to investigate the neurophysiological correlates of various meditation forms, including religious chanting. This method uniquely integrates fMRI eigenvector results for region selection in electroencephalogram (EEG) source analysis using k-means clustering. The results provide an in-depth understanding of the neural processes involved in repetitive religious chanting.

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Medicine

Examination of Oral Candida Infection in Primary Sjögren's Syndrome Patients
Xiaoyan Wang 1,2,3, Yeqing Song 4, Xinzhe Lou 1, Bingqing Han 5,6,7, Ruiqing Wu 1, Jing Xie 8,9, Xiang Lin 8,9, Hao Wang 1
1Department of Stomatology, Beijing Tiantan Hospital, Capital Medical University, 2Beijing Laboratory of Oral Health, Capital Medical University, 3Department of Biochemistry and Molecular Biology, School of Basic Medicine, Capital Medical University, 4Central Laboratory, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, 5Clinical Diagnosis Laboratory of Beijing Tiantan Hospital, Capital Medical University, 6Key Laboratory for Quality Control of In Vitro Diagnostics, NMPA, 7Beijing Engineering Research Center of Immunological Reagents Clinical Research, 8School of Chinese Medicine, The University of Hong Kong, 9Department of Rheumatology and Clinical Immunology, Beijing Tiantan Hospital, Capital Medical University

Here, we present a protocol for the examination of oral Candida infection in patients with primary Sjögren's syndrome, which can be used for timely treatment and, thereafter, avoiding related complications.

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Medicine

Laparoscopic Non-Mesh Cerclage Pectopexy with Uterine Preservation for Pelvic Organ Prolapse
Chaoxia Lyu *1, Willy Cecilia Cheon *2, Hextan Yuen Sheung Ngan *3, Yuzhen Wei 1, Wenju Zhang 1
1Department of Gynecology and Obstetrics, The University of Hong Kong-Shenzhen Hospital, 2Department of Gynecology and Obstetrics, Queen Elizabeth Hospital, 3Department of Obstetrics and Gynecology, Queen Mary Hospital, The University of Hong Kong

Here we introduce a novel approach known as laparoscopic non-mesh cerclage pectopexy with uterine preservation for pelvic organ prolapse (LNMCPUP) for patients who wish to preserve their uterus. Permanent cerclage sutures are used to suspend the uterus to the iliopectineal ligament through the round ligament.

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