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The Hong Kong Polytechnic University

8 ARTICLES PUBLISHED IN JoVE

Biology

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Kenneth K. Kwong^1,2, Lenuta Kloetzer^1,2,3,4, Kelvin K. Wong^5,6, Jia-Qian Ren^1,2, Braden Kuo^1,2,3,4, Yan Jiang⁷, Y. Iris Chen^1,2, Suk-Tak Chan^1,2,8, Geoffrey S. Young⁹, Stephen T.C. Wong^5,6

¹Department of Radiology, Massachusetts General Hospital, Harvard Medical School, ²Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, ³Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, ⁴Department of Medicine, Massachusetts General Hospital, Harvard Medical School, ⁵Center for biotechnology and Informatics, The Methodist Hospital Research Institute, ⁶Department of Radiology, The Methodist Hospital, Weill Cornell Medical College, ⁷Bejing University of Chinese Medicine, ⁸Department of Health Technology and Informatics, The Hong Kong Polytechnic University, ⁹Department of Radiology, Brigham and Women's Hospital, Harvard Medical School

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

Genetics

In Vitro Transcription Assays and Their Application in Drug Discovery

Xiao Yang¹, Cong Ma^1,2

¹School of Environmental and Life Sciences, University of Newcastle, ²Department of Applied Biology and Chemical Technology, The State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University

In this manuscript, we describe a protocol to functionally examine transcription and the inhibitory activity of antibacterial agents targeting bacterial transcription.

Medicine

Minimally Invasive Transverse Aortic Constriction in Mice

Aung Moe Zaw¹, Connor M. Williams², Helen K. W. Law³, Billy Kwok Chong Chow¹

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.

Neuroscience

Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture

Lin Jiajia^*1, Mak Shinghung^*2, Zheng Jiacheng¹, Wang Jialing¹, Xu Dilin¹, Hu Shengquan², Zhang Zaijun³, Wang Qinwen¹, Han Yifan^2,4, Cui Wei^1,2

¹Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, ²Department of Applied Biology and Chemistry Technology, Institute of Modern Chinese Medicine, The Hong Kong Polytechnic University, ³Institute of New Drug Research, Guangdong Provincial Key Laboratory of Pharmacodynamic, Constituents of Traditional Chinese Medicine & New Drug Research, College of Pharmacy, Jinan University, ⁴International Joint Laboratory (SYSU-PolyU HK) of Novel Anti-Dementia Drugs of Guangdong

This protocol describes how to accurately measure neuronal viability using Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining in cultured cerebellar granule neurons, a primary neuronal culture used as an in vitro model in neuroscience and neuropharmacology research.

Biochemistry

A11-positive β-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis

Huang Chunhui^1,2, Xu Dilin¹, Zhang Ke¹, Shentu Jieyi², Yan Sicheng¹, Wu Dapeng³, Wang Qinwen¹, Cui Wei^1,2

¹Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, ²Li Dak Sum Yip Yio Chin Kenneth Li Marine Biopharmaceutical Research Center, Ningbo University, ³School of Materials Science and Chemical Engineering, Ningbo University

This protocol describes how to prepare Aβ oligomers from a synthetic peptide in vitro and to evaluate relative amounts of Aβ oligomer by a dot blotting analysis.

Neuroscience

A Pilot Study on the Repetitive Transcranial Magnetic Stimulation of Aβ and Tau Levels in Rhesus Monkey Cerebrospinal Fluid

Ling-Yi Liao^1,2,3, Ying-Qian Zhang⁴, Benson Wui-Man Lau⁵, Qiong Wu⁶, Zi-Yan Fan^1,3, Qiang Gao^1,3, Zhi-Hui Zhong^6,7

¹Department of Rehabilitation Medicine, West China Hospital, Sichuan University, ²Department of Rehabilitation Medicine, Daping Hospital, Army Medical University, ³Key Laboratory of Rehabilitation Medicine in Sichuan Province, West China Hospital, Sichuan University, ⁴Department of physiology, Southwest Medical University, ⁵Department of Rehabilitation Sciences, The Hong Kong Polytechnic University, ⁶Laboratory of Nonhuman Primate Disease Modeling Research, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, ⁷Sichuan Kangcheng Biotech Co., Inc.

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

Medicine

A Protein Suspension-Trapping Sample Preparation for Tear Proteomics by Liquid Chromatography-Tandem Mass Spectrometry

Jimmy Sung-Hei Tse^*1, Ying-Hon Sze^*1,2, Jimmy Ka-Wai Cheung^1,2, King-Kit Li¹, Thomas Chuen Lam^1,2,3,4

¹Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, ²Centre for Eye and Vision Research (CEVR), ³Research Centre for SHARP Vision (RCSV), The Hong Kong Polytechnic University, ⁴Research Centre for Chinese Medicine Innovation (RCMI), The Hong Kong Polytechnic University

This protocol describes a method for collecting tear samples using Schirmer strips and an integrated quantitative workflow for discovering non-invasive tear protein biomarkers. The suspension trapping sample preparation workflow enables fast and robust tear sample preparation and mass spectrometric analysis, resulting in higher peptide recovery yields and protein identification than standard in-solution procedures.

Neuroscience

Simultaneous Focused Ultrasound Neuromodulation and Fiber Photometry Recording in Free-Moving Mouse

Na Li^*1,2, Zihao Chen^*1,3, Jiejun Zhu¹, Hanming Zheng^1,4, Jinzhao Xia^1,5, Zhen Yuan⁴, Chunlong Fei², Lei Sun³, Zhihai Qiu¹

¹Guangdong Institute of Intelligence Science and Technology, ²School of Microelectronics, Xidian University, ³Department of Biomedical Engineering, The Hong Kong Polytechnic University, ⁴Centre for Cognitive and Brain Sciences and Ministry of Education Frontiers Science Center for Precision Oncology, University of Macau, ⁵Deep Brain Technology Co., Ltd

The protocol includes transducer manufacturing, parameters reporting, surgical procedure, and signal recording for the entire operational workflow of concurrent focused ultrasound neuromodulation and fiber photometry recording in free-moving mice.