The experimental study of rAAV2 transfecting neural stem cells-derived neurospheres / 中国应用生理学杂志

<p><b>AIM</b>To investigate the abilities of recombinant adeno-associated virus type 2 (rAAV2) transfecting neurospheres.</p><p><b>METHODS</b>The rAAV2 conjugated with FITC (rAAV2-FITC) was added into the culture medium of neurospheres and 30 minutes later the neurospheres were detected with a fluorescence microscopy to determine if the AAV can combine with neurospheres. The rAAV2 containing GFP reporter gene (rAAV2-GFP) was incubated with the neurospheres for a month and then detected the ability of transfecting neurospheres. The neurospheres transfected with rAAV2-containing GFP were transplanted to the brain of rats. A month later the rats were sacrificed and the brains were removed to detect if there are expressions of the reporter gene. The neurospheres were transfected with rAAV2 containing hypoxia responds elements (HRE) and vascular endothelium growth factor(VEGF) gene and reporter gene GFP (rAAV2-HRE-VEGF-GFP) and then cultured in low oxygen density environments. Seventy-two hours later the neurospheres were detected through a fluorescence microscopy.</p><p><b>RESULTS</b>The neurospheres incubated with rAAV2-FITC present bright green fluorescence. GFP, the reporter gene, can be seen clearly 1 month after being transfected with rAAV2-GFP. The same green fluorescence protein can be observed ex vivo as well. The fluorescence can be seen in neurospheres transfected by rAAV2-HREVEGF-GFP only in low oxygen density.</p><p><b>CONCLUSION</b>The rAAV2 can transfect neurospheres specifically and efficiently. Reporter gene can be expressed in the neurospheres in vivo and ex vivo. Expression of reporter gene can be adjusted by HRE.</p>

Effects of scorpion venom active peptides on the concentration of PGI_2 and NO secreated by human umbilical vein endothelical cells / 中国临床药理学与治疗学

Aim To further research into the antithrombotic mechanism of scorpion venom active peptides (SVAP). Methods Human umbilical vein endothelial cells (HUVEC) were cultured with enzyme digestive method. After the cultured HUVC was incubated in conditioned media for 1 hour, the effects of SVAP on the concentration of 6-Keto-PGF 1? and NO of HUVEC were determined with radioactive-immunolygic and nitrate reduction enzyme method respectively. Results As compared with control, SVAP in the doses of 1,5,10, 20 mg?L -1 had the distinctive increase of 54%, 68%,72%,79% of the concentration of 6-Keto-PGF 1? and SVAP in the doses of 10, 20 mg?L -1 had the significantly increased of 27%, 46% of the concentration of NO. Regression anylysis showed that the release levels of PGI 2 and NO in HUVEC induced by SVAP was of positive correlation. Conclusion Antithrombotic mechanism of SVAP is related to the increase of PGI 2 and NO released from HUVEC and synergistic and mediating action between NO and PGI 2.