The Influences On Lung Adenocarcinoma-related Staphylococcus Epidermidis In The Formation Of Biofilm By Accumulation Associated Protein And Transform Growth Factor-beta1

(Objective] To study the Influences on Lung adenocarcinoma-related Staphylococcus epidermidis in the formation of biofilm by Accumulation Associated Protein and Transform growth factor-beta1[Method] In this experiment:1. The cultivation of human lung adenocarcinoma A549cells in vitro, the stable after the wedding, centrifugal obtain its supernatant fluid, the supernatant fluid and different concentrations of transforming growth factor beta1(0,10,20,40ng/ml) work together to separated from human malignant pleural effusion mononuclear cells, after24hours, by flow cytometry instrument measure the levels of each related immune factors.2. Separation of non-small cell lung cancer patients chest drainage tube staphylococcus epidermisdis clinical strains of bacterial DNA extraction, PCR tests related protein (Aap) gene, establishing stable genetic aggregated proteins associated with negative and positive isolates.3. Use medical silicone rubber with46strains of Staphylococcus epidermidis clinical isolates cultured in a medium for30hours and test the bacterial biofilm formation by semi-quantitative adhesion measured strains.4. Take the train for24hours after four different concentrations of transforming growth factor beta1set of common culture supernatant, join epidermis staphylococcus genetic aggregated proteins associated with negative, positive standard strains and the staphylococcus epidermisdis aggregated proteins related to negative, positive clinical isolates, and to join the medical silicon rubber as a biological material, joint training after30hours. take out the biological materials, via a semi-quantitative test each bacterial biofilm formation, bacterial biofilm surface is observed by scan electron microscope. The results of the experiment using SPSS17.0statistical software for statistical processing.[Results]1. Under different concentrations of TGF-beta1factors stimulation,30hours after the flow cytometry instrument to detect CD3+, CD4+CD8+cell ratio in the10ng/ml group,20ng/ml group,40ng/ml group is lower than the blank control group(p<0.01); Through q test found CD3+, CD4+, CD8+cell ratio and TGF-beta1factor is negatively related to the effect dose relationship (p<0.05). CD4+/CD8+, CD4+CD25+cells percentage in the10ng/ml group,20ng/ml group,40 ng/ml group than in blank control group (p<0.01), by q Test found that CD4+/CD8+, CD4+CD25+cells percentage and TGF-beta1factor is positively related to the dose effect relationship (p<0.05). In the10ng/ml group,20ng/ml group and40in the ng/ml group CD3+CD25+cells percentage is lower than the blank control group, through the inspection found no means and TGF-beta1factor showed effect relationship (p>0.05).2different concentration TGF-beta factorl, cells after30hours after the inspection found that IL-2and TNF-a, IFN-gamma density in the10ng/ml group,20ng/ml group and40in ng/ml group is lower than the blank control group (p<0.01); Through q inspection found that IL-2and TNF-a, IFN-gamma concentration and TGF-beta1factor is negatively related to the dose effect relationship (p<0.05).Il-6concentration in the10ng/ml group, and20ng/ml group,40ng/ml group than in blank control group (p<0.01), by q inspection found that il-6concentrations and TGF-beta1factor is positively related to the dose effect relationship (p<0.05). Concentration of IL4, IL-10through the inspection found no q means and TGF-beta1factor showed dose effect relationship (p>0.05).3. Medical silicone rubber and Staphylococcus epidermidis aap gene negative, positive clinical isolates grown in culture30hours, the semi-quantitative adhesion test Staphylococcus epidermidis aap gene negative, positive clinical isolates film-forming: Fisher’s exact test foundthe the Aap genes exist biofilm formation is closely related (p <0.01), the Aap genes play a role in the formation of bacterial biofilms.4. Medical silicone rubber and the staphylococcus epidermisdis standard strains,6strains of staphylococcus epidermis Aap positive clinical isolates out weeks mononuclear cells and A549lung adenocarcinoma cancer cell supernatant TGF-beta1factors stimulation at different concentration (0ng/ml,10ng/ml, and20ng/ml.40ng/ml) under common30hours of training by semi-quantitative method measure results:10ng/ml, and20ng/ml,40ng/mlTGF-beta1factor stimulating SE986, SE842, SE317, SE276, RP62A bacterial biofilm thickness is greater than the blank control group (p<0.01). Through the epidermis staphylococcus group q inspection found that the thickness of the bacterial biofilm and TGF-beta1factor level is positively related top<0.05) in different TGF-beta1factor level set of bacterial biofilm thickness no correlation (p>0.05).standard of medical silicone rubber and the staphylococcus epidermisdis. Takes6strains of staphylococcus epidermis Aap clinical isolates weeks out negative mononuclear cells and A549lung adenocarcinoma cancer cell supernatant TGF-beta1factors stimulation at different concentration (0ng/ml,10ng/ml, and20ng/ml,40ng/ml) under30hours of training together by semi-quantitative method measurement results:10ng/ml, and20ng/ml.40ng/mlTGF-beta1factor under the stimulus of the thickness of epidermis staphylococcus and blank control group no difference(p>0.05). Through various levels TGF-beta1is no difference between groups (p>0.05). Experimental proof that keeps Aap negative TGF-beta1factors stimulation at different concentration (0ng/ml,10ng/ml. and20ng/ml,40ng/ml) were not formation of bacterial biofilm.[Conclusion]1..Transforming growth factor beta inhibit activation of T lymphocytes for lung cancer patients, cells decreased immune function. Cause the Thl/Th2balance disorders, leading to Thl-driven cellular immune function is suppressed.2. The App gene is closely related to biofilm formation。The Aap genes play a role in the formation of bacterial biofilms3.Transforming growth factor beta the agrin gene positive epidermal aureus formed the bacterial biofilms role, for the the agrin negative epidermal aureus had no effect on the formation of bacterial biofilms.