Army Medical University (Third Military Medical University)

5 ARTICLES PUBLISHED IN JoVE

Biology

Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

Qianfan Bai^*1, Zhiqiang Bai^*1, Shaohai Xu³, Lei Sun^1,2

¹Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, ²Institute of Molecular and Cell Biology, Singapore, ³Division of Bioengineering, School of Chemical & Biomedical Engineering, Nanyang Technological University

An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.

Cancer Research

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

Jenny Lazarus¹, Yagiz Akiska¹, Mirna Perusina Lanfranca¹, Lawrence Delrosario¹, Lei Sun¹, Daniel Long², Jiaqi Shi³, Howard Crawford², Marina P. Di Magliano¹, Weiping Zou¹, Timothy Frankel¹

¹Department of Surgery, University of Michigan, ²Department of Molecular and Cellular Physiology, University of Michigan, ³Department of Pathology, University of Michigan

Multiplex fluorescent immunohistochemistry is an emerging technology that enables the visualization of multiple cell types within intact formalin-fixed, paraffin embedded (FFPE) tissue. Presented are guidelines for ensuring a successful 7-color multiplex with instructions for optimizing antibodies and reagents, preparing slides, design and tips for avoiding common problems.

Genetics

Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator

Kayli Sieber¹, Maya Saar¹, Comzit Opachaloemphan², Matthew Gallitto³, Huan Yang², Hua Yan^1,4

¹Department of Biology, University of Florida, ²Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, ³Department of Radiation Oncology, Columbia University Medical Center, ⁴Center for Smell and Taste, University of Florida

Many characteristics of insect eusociality rely on within-colony communication and division of labor. Genetic manipulation of key regulatory genes in ant embryos via microinjection and CRISPR-mediated mutagenesis provides insights into the nature of altruistic behavior in eusocial insects.

Medicine

Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects

Xi Ling¹, Peng Zou¹, Lin Ao¹, Niya Zhou¹, Xiaogang Wang², Lei Sun¹, Huan Yang¹, Jinyi Liu¹, Jia Cao¹, Qing Chen¹

¹Institute of Toxicology, College of Preventive Medicine, Army Medical University (Third Military Medical University), ²Department of Chemical Defense Medicine, College of Military Preventive Medicine, Army Medical University (Third Military Medical University)

The present protocol provides a practical solution that allows for the measurement of apoptosis, mitochondrial membrane potential, and DNA damage in human sperm with a single cytometer.

Medicine

Transoral Robotic Total Thyroidectomy and Bilateral Central Regional Lymph Node Dissection for Papillary Thyroid Carcinoma

Shu Zhang¹, Gang Zhang¹, Jing Xu¹, Yan Jiang¹, Yan Xu¹

¹Department of Breast and Thyroid Surgery, Daping Hospital, Army Medical University (Third Military Medical University)

The transoral robotic total thyroidectomy and bilateral central regional lymph node dissection using three ports without axillary incision are feasible and safe for early-stage papillary thyroid carcinoma. Here we present the operative techniques of this operation.