Effect of Yes-associated protein in regulation of electroacupuncture pretreatment on cerebral ischemia reperfusion injury rats / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) pretreatment on inflammatory reaction, apoptosis and expression of Yes-associated protein (YAP) of ischemic penumbra of cerebral cortex in cerebral ischemia reperfusion injury rats, and to explore the possible mechanism of its neuroprotection effect.@*METHODS@#A total of 84 SD rats were randomized into a sham operation group (12 rats), a model group (18 rats), an EA group (18 rats), an EA+YAP virus transfection group (18 rats) and an EA+virus control group (18 rats). Except for the sham operation group, thread embolization method was adopted to establish the middle cerebral artery occlusion (MCAO) model in rats of the other groups. EA was applied at "Baihui" (GV 20) and "Dazhui" (GV 14) for 30 min in the 3 EA intervention groups 2 h before model establishment, disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in intensity. Adenovirus transfection technique was used to induce gene silencing of YAP in the EA+YAP virus transfection group, and adenovirus vectors was injected as negative control in the EA+virus control group 4 d before model establishment. Twenty-four hours after model establishment, neurological function score was evaluated, the relative cerebral infarction area was observed by TTC staining, the apoptosis in the ischemic penumbra of cerebral cortex was detected by TUNEL staining, the levels of inflammatory factors IL-1β, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex was detected by ELISA method, the expression of YAP was detected by Western blot and immunofluorescence.@*RESULTS@#Compared with the sham operation group, the expression of YAP was increased in the model group (@*CONCLUSION@#Electroacupuncture pretreatment can effectively improve the ischemia reperfusion injury, its mechanism may be related to up-regulating the expression of YAP in the ischemic penumbra of cerebral cortex and relieving the apoptosis and inflammatory reaction.

Development of microfluidic chip with adjustable concentration and pressure gradient for 3D cell culture / 医用生物力学

Objective To develop a microfluidic device with the adjustable concentration and pressure gradient for 3D cell culture in hydrogel and set up an in vitro model with the capability to closely simulate in vivo microenvironment for cell growth. Methods The microfluidic chip, with a middle channel for 3D cell culture and two side channels for delivering cell culture medium, was designed and fabricated using standard soft lithography and replica molding techniques. Its capability to generate concentration gradient, interstitial flow and image cell in situ was demonstrated. Results A simple microfluidic chip for 3D cell culture in hydrogel with the capability to generate the concentration and pressure gradient was obtained. At a flow rate of 2 μL•min-1 in each side channel, the concentration gradients remained constant after 3 h. The interstitial flow across the gel scaffold was generated by a 100 Pa pressure difference between two-side channels with the pressure gradient of 0.11 Pa/μm. Human adult dermal microvascular endothelial cells (HMVEC) were maintained in 3D culture with collagen type I and observed with confocal microscopy. Conclusions The microfluidic chip is simple and easy to operate and it can simulate the complicated microenvironment in vivo. The chip also allows the multiparameter control of microenvironment, facilitating the better understanding of interaction between cells and microenvironment.

Expression of HPV16 L1 protein in insect cell suspension culture system / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the L1 protein of human papillomavirus type 16 (HPV16) in insect cell suspension culture system.</p><p><b>METHODS</b>Optimized the conditions of suspension culture, recombinant virus amplification and protein expression. Determined the virus tilter by plague analysis and detected the target protein by SDS-PAGE and Western blot; The formation of VLPs by HPV16 L1 protein was observed with TEM.</p><p><b>RESULTS</b>The Sf9 cells could grow better in suspension culture with seeding density of 5 x 10-5 cell/mL and the maximum expression quantity was obtained by infection of cells with rBacV/HPV16L1 (MOI =10) and harvesting after 72-84 h. HPV16L1 protein could assemble into VLPs in Sf9 cells observed with TEM.</p><p><b>CONCLUSION</b>The conditions of cell culture, virus amplification and protein expression were optimized. HPV16 L1 protein could assemble into VLPs in Sf9 cells, which would provide a foundation for further study of the vaccine and diagnosis kits.</p>

Analysis of HBV precore 1896 site mutation and its HBV genotype and other laboratory features / 第二军医大学学报

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P