Primary culture and identification of rat glomerular microvascular endothelial cells / 生理学报

The aim of the present study was to establish a simple and efficient method for isolation and culture of primary rat glomerular microvascular endothelial cells in vitro. The bilateral kidneys were taken from 7-10-day old Sprague-Dawley rats, and the renal cortex was separated. Glomeruli were obtained by cutting and continuously passing 200-mesh and 300-mesh sieves. After type IV collagenase digestion for 15-20 min, renal microvascular globules were collected for inoculation and culture. The cultured cells were identified by cell morphology observation and immunocytochemical staining with factor VIII related antigen. The results showed that the renal microvascular globules were irregularly spherical, without cysts, and the capillary loop structure was clear; after 3 days of primary culture, short spindle-shaped cells crawled out around the renal microvascular globules and gradually formed cell colonies, showing an "island-like shape" distribution; 4-5 days later, the cell colonies fused with each other; 6 days later, the cells covered the bottom of the dish, showing a typical monolayer, paving stone-like, mosaic arrangement. The immunocytochemical staining of factor VIII related antigen showed that the cytoplasm was lightly stained brownish red, and factor VIII related antigen-positive rate of cells was nearly 100%. The above results suggested that this study successfully established a method combining continuous screening and collagenase digestion for culture of primary rat glomerular microvascular endothelial cells in vitro. It provides an important tool cell for studying the mechanism of the occurrence and development of diabetic nephropathy.

Improvement on the methods about the primary culture of rat brain microvascular endothelial cells and identification / 解剖学报

Objective To improve the method about the primary culture of rat brain microvascular endothelial cells in vitro. Methods The SD rats aged from 4 to 6 weeks were chosen as research object. After craniotomy, washing and cutting, sieving, density gradient concentration of BSA, digestion of type II collagenase and collagenase dispersive enzyme twice, the primary culture was carried out. The target cells were indentified by morphological abservation and immunocytochemical staining of facter VI. Results Cultured for 12 to 24 hours,the cells in vitro migrated outward from the microvascular section. The cells appeared polygonal-shaped, and proliferated in a clustered monolayer, the cell growth density reached 70% - 80% of the bottle bottom after 3 days, and arranged like cobbles. The correlation antigen of VI factor was positive, they reached confluence with over purity 99%. Conclusion The method is available that can successfully separate and cultivate microvascular endothelial cells of rat brains in vitro.

Research progress in roles of high-risk human papillomavirus E2 protein / 病毒学报

High-risk human papillomavirus (HPV) is the principal cause of various cancers including cervical cancer, anal cancer, vulvar cancer, and some head and neck cancers. In the viral life cycle, by interacting with both viral and host DNA and proteins, the HPV E2 protein plays a pivotal role in viral transcriptional regulation and DNA replication, and it is also associated with modification of various cellular processes, including host gene transcription, RNA processing, apoptosis, ubiquitination, and intracellular trafficking, to create a convenient environment for a replicative cycle of the virus and contribute to the HPV pathogenesis. Elucidating the roles of E2 protein throughout the viral life cycle will improve our understanding of the viral life cycle and pathogenesis and help us identify novel antiviral agents with therapeutic potential. This article reviews the research progress in the structure, roles, and activity of high-risk HPV E2 protein, particularly that of HPV-16.