Effect of propofol on interleukin-1β-induced increase in monolayer permeability of human umbilical vein endothelial cells / 中华麻醉学杂志

Objective To evaluate the effect of propofol on interleukin-1β (IL-1β)-induced increase in monolayer permeability of human umbilical vein endothelial cells (HUVECs).Methods Primary HUVECs were cultured and purified by immuno-magnetic separation.The expression of VE-cadherin in endothelial cells was determined by immunofluorescence.The HUVEC monolayer permeability was detected by the Transwell system.The cells were seeded on the upper chamber (2 × 105 cells/well) and cultured for 3 days after confluence.The cells were treated in two ways.The cells were randomly divided into 6 groups (n =36 each) and 5 of the 6 groups treated with 1,2,5,10 and 20 ng/ml IL-1β for 24 h except for control group.The cells were also randomly divided into 5 groups (n =30 each) and 4 of the 5 groups were pretreated with 0,10,50 and 100 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h except for control group.The cells were radomly divided into 3 groups (n =18 each) and 2 of the 3 groups were pretreated with 50 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h or 30 min.The expression of occludin protien,p38 mitogen activiated protienkinase (p38 MAPK) and phosphorylated p38 MAPK (p-p38 MAPK) was determined by Western blot.Results Compared with control group,5,10 and 20 ng/ml IL-1β significantly increased HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.05 or 0.01).10,50 and 100 μmol/L propofol inhibited IL-1 β-induced increase in the permeability of HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.01).IL-1β could down-regulate HUVEC occludin protein expression,and activate p38MAPK signaling pathway,and propofol inhibited IL-1β-induced down-regulation of HUVEC occludin protein expression and activation of p38 MAPK signaling pathway (P ＜ 0.01).Conclusion Propofol can alleviate IL-1β-induced increase in the permeability of HUVEC monolayer via inhibiting activation of p38 MAPK signaling pathway.

A rapid and simple method for extracting mitochondrial DNA from human lung cancer tissue / 中国肺癌杂志

<p><b>BACKGROUND</b>The relationship between mitochondrial DNA (mtDNA) mutation and tumor is the hot point in cancer research field by now. The existent methods for extracting mtDNA are time-consuming and complicated. In order to further research the relationship of the mutation of mtDNA and development of lung cancer, it is necessary to establish a rapid and simple method for extracting mtDNA from lung cancer tissue.</p><p><b>METHODS</b>Lung cancer tissues were lysed in base-denaturalization liquid and mtDNA was directly extracted from them by chloroform:isoamyl alcohol (24:1). The mtDNA was confirmed by PCR.</p><p><b>RESULTS</b>The extracted mtDNA samples were not combinated with protein and 1528bp PCR products of (mtDNA) were detected from 40 samples of analyzing patients. About (7.97±0.12)μg of mtDNA could be extracted from 1 gram of cancer tissue.</p><p><b>CONCLUSIONS</b>The method is simple, rapid and efficient to extract (mtDNA) from human lung cancer tissues and it could be used to handle small amount tissues or large number of samples in clinical and scientific research.</p>