Platycodin D protects oral epithelial cells against infection of Candida albi-cans / 中国病理生理杂志

AIM: To investigate the effect of platycodin D on Candida albicans infection in oral epithelial cells.METHODS:The viability of the oral squamous carcinoma KB cells was detected by MTT assay after treated with different concentrations of platycodin D .The KB cells were infected with Candida albicans, and then were incubated with platycodin D at different concentrations .Adherent numbers of the Candida albicans were counted by Gram staining , and the bacterial activity and conversion were measured by Trypan blue staining .Furthermore , the protein levels of IL-18 and hu-man β-defensin 2 ( HBD-2) were analyzed by ELISA , and the expression of HBD-2 at mRNA and protein levels was deter-mined by RT-qPCR and Western blot , respectively .RESULTS:The viability of the KB cells was not affected by platycod-in D at the concentrations used .The adherent numbers , bacterial activity and conversion were decreased by treatment with platycodin D in a dose-dependent manner .In addition, the protein level of IL-18 in the culture supernatant and the mRNA expression of HBD-2 in the KB cells were also reduced after platycodin D treatment .CONCLUSION:Platycodin D has a bacteriostasis effect and prevents oral epithelial cells from Candida albicans infection.

Construction of three lentiviral vectors expressing HPV16 E5, E6 or E7 oncogene and respective infection of oral epithelial cells with the vectors / 安徽医科大学学报

Objective To provide the basis for the study of HPV16 E5 gene inthe pathogenesis of oral squamous cell carcinoma by constructing three lentiviral vectors expressing HPV 16 E5, E6 or E7 oncogene and transfecting o-ral epithelial cells with the vectors.Methods HPV16 E5, E6 or E7 oncogene was amplified using PCR and liga -ted into the lentiviral vector pLVX-AcGFP-N1 respectively to construct vectors pLVX -AcGFP-E5, pLVX-AcGFP-E6 and pLVX-AcGFP-E7, then respectively cotransfected 293T cells with packaging plasmids , viral supernatant was collected to respectively transfect oral epithelial cells .Afterpuromycin screening,oral epithelial cells with HPV 16 E5, E6,or E7 oncogene transfection were constructed , then reverse transcription PCR and western blotassays were performed for verifying HPV16 E5, E6 or E7 expression.Results pLVX-AcGFP-E5, pLVX-AcGFP-E6 and pLVX-AcGFP-E7 were successfully constructed, oral epithelial cells expressing HPV 16 E5, E6 or E7 oncogene wereacquired, HPV16 E5,E6 or E7 expression was confirmed in oral epithelial cellsthrough reverse transcription PCR and western blot assays.Conclusion Three lentiviral vectors expressing HPV16 E5, E6 or E7 oncogene can successfully infect oral epithelial cells .

Peptidoglycan Induces the Production of Interleukin-8 via Calcium Signaling in Human Gingival Epithelium

The etiology of periodontal disease is multifactorial. Exogenous stimuli such as bacterial pathogens can interact with toll-like receptors to activate intracellular calcium signaling in gingival epithelium and other tissues. The triggering of calcium signaling induces the secretion of pro-inflammatory cytokines such as interleukin-8 as part of the inflammatory response; however, the exact mechanism of calcium signaling induced by bacterial toxins when gingival epithelial cells are exposed to pathogens is unclear. Here, we investigate calcium signaling induced by bacteria and expression of inflammatory cytokines in human gingival epithelial cells. We found that peptidoglycan, a constituent of gram-positive bacteria and an agonist of toll-like receptor 2, increases intracellular calcium in a concentration-dependent manner. Peptidoglycan-induced calcium signaling was abolished by treatment with blockers of phospholipase C (U73122), inositol 1,4,5-trisphosphate receptors, indicating the release of calcium from intracellular calcium stores. Peptidoglycan-mediated interleukin-8 expression was blocked by U73122 and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). Moreover, interleukin-8 expression was induced by thapsigargin, a selective inhibitor of the sarco/endoplasmic reticulum calcium ATPase, when thapsigargin was treated alone or co-treated with peptidoglycan. These results suggest that the gram-positive bacterial toxin peptidoglycan induces calcium signaling via the phospholipase C/inositol 1,4,5-trisphosphate pathway, and that increased interleukin-8 expression is mediated by intracellular calcium levels in human gingival epithelial cells.

The exploration of STR genotyping of minute cells by MiniFiler kit / 中国法医学杂志

Objective Explore the feasibility of minute cells STR typing.Methods Minute cells which were obtained by micromanipulation method were amplified with MiniFiler kit,and detected by ABI 3130 genetic analyzer.Results Ten cells can be successfully genotyped.Complete genotyping results can be obtained for one,three or five cells but with stochastic effect.Conclusion Instability is observed in minute cells genotyping,so it can not be used in actual case work.Maybe increase the quantity of DNA template can improve the success rate.

Effect of Candida albicans on cell cycle distribution of KB cells / 实用口腔医学杂志

Objective:To investigate the effects of Candida albicans(C. albicans) on the cell cycle distribution of oral epithelial KB cells in vitro. Methods:KB cells at 3?106/ml were seeded and cultured for 48 h, then the bacterial filament and the spore of C. albicans at 102 CFU/ml were added to the cultures respectively. KB cell culture without C. albicans was used as the control. After 48 h culture cell suspensions were prepared and subjected to FACS flow cytometry for detecting the cell cycle distribution. Results:Filament of C. albicans decreased the percentage of G0/G1 cells, increased G2/M cell population and proliferation index of the cells(P