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Objective To investigate the effect of Dectin-1 on the release of inflammatory factors interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in rat tracheal epithelial cells (RTECs) stimulated with heat-treated Candida glabrata (C.glabrata).Methods RTECs cultivated in vitro were randomly divided into three groups,including control group(RTECs + sterile normal saline),fungal stimulation group(RTECs + heat-treated C.glabrata),and inhibitor intervention group(RTECs + laminarin + heat-treated C.glabrata),cells were harvested after incubation for 0,2,4,6 hours respectively,cell survival rate was determined by MTT method,expression of Dectin-1 was analyzed by Western Blot,expression of IL-10 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).Results Heat-treated C.glabrata destroyed cell structure and reduced cell survival rate.At the beginning of the culture (0 h),cell survival rate,expressions of Dectin-1,IL-10 and TNF-α among three groups were all not significantly different(all P＞0.05).After incubation for 2,4,6 hours,expressions of Dectin-1,IL-10 and TNF-α in fungal stimulation group and inhibitor intervention group were both significantly higher than control group;expressions of Dectin-1,IL-10,and TNF-α in inhibitor intervention group was lower than fungal stimulation group(all P＜0.05).The expressions of IL-10 in inhibitor intervention group at 0 h and 2 h was not significantly different,expressions of Dectin-1,IL-10,and TNF-α in fungal stimulation group and inhibitor intervention group at different incubation periods were significantly different(all P＜0.05).Conclusion Dectin-1 is an important receptor for RTECs to recognize the heat-treated C.glabrata,it induces the release of IL-10 and TNF-α,and mediate the occurrence of inflammation.
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Objective To explore the effects of co‐cultured heat‐treated candida glabrata with rat tracheal epithelial (RTE) cells on the expression of Dectin‐1 and the production of IL‐6 and TNF‐α.Methods RTE cells in vitro were co‐cultured with heat‐treated candida glabrata bacteria liquid for 2 ,4 ,6 h ,while without co‐cultured RTE cells were used as control group .We observed the morphological changes of RTE cells ,detected the protein expressions of Dectin‐1 by Western blot ,used real‐time PCR to detecte the mRNA expressions of IL‐6 and TNF‐αand measured protein expression of IL‐6 and TNF‐αby ELISA .Results With the pass‐ing of time ,the RTE cells were damaged extensively and the expression of Dectin‐1 ,IL‐6 and TNF‐αbecame more and more signifi‐cant .Obviously ,there had significant difference in the expression of Dectin‐1 ,IL‐6 and TNF‐α between the co‐cultured 2 h group and the control group ,the co‐cultured 4 h group and the co‐cultured 2 h group ,the co‐cultured 6 h group and the co‐cultured 4 h group (P<0 .05) .Conclusion RTE cells have natural immune function .The Dectin‐1 involves in the recognition of heat‐treated Candida glabrata ,activating secretion of IL‐6 and TNF‐αand mediating inflammatory reaction .IL‐6 plays a negative regulation role .
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AIM:To investigate the role of phospholipase C epsilon 1 ( PLCE1 ) in modulating the apoptotic mechanism in lung adenocarcinoma A 549 cells.METHODS:PLCE1 inhibitor U-73122 was used to suppress the expres-sion of PLCE1.The expression of PLCE1 and p53 in A549 cells was evaluated by quantitative real-time PCR and Western blotting.Apoptosis was assessed by flow cytometry .RESULTS:A549 cells expressed high level of PLCE1 and low level of p53.Inhibition of PLCE1 markedly increased the expression of p 53, and increased the apoptosis of A 549 cells.CON-CLUSION:PLCE1 suppresses apoptosis of A549 cells via inhibiting the expression of p53.
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Objective To study the roles of Toll-like receptor 2(TLR2)and Toll-like receptor 4(TLR4)in the progress of pulmo-nary infection with candida glabrata in experimental mice .Methods The mice were divided into three groups ,the control group (group A) ,mice infected with candida glabrata(group B)and immunosuppressive mice infected with candida glabrata(group C) .6 mice in each group were killed at 1st and 3rd day after the success in pulmonary infection with candida glabrata .The lung tissues from each group were collected for pathological analysis and PT-PCR to detect the expression level of TLR2 and TLR4 .Protein TNF alpha(TNF-α)level was measured by ELISA method .Results The pathological analysis showed the structure was normal and there was no inflammatory reaction in lungs in group A .The infiltration of inflammatory cells ,weak injuries but no conidia or hyphae in lungs were found in group B .Most of the alveolar collapse and severe damage ,part of the expansion ,a large number of in-flammatory cells infiltration ,accompanying with conidia and hyphae accumulation were observed in group C .The expression levels of TLR2 mRNA and TNF-αprotein in mice lungs at the 1st day and the 3rd day in group C were significantly higher than in group A and B(P<0 .05) ,while group B was higher than group A(P<0 .05);The expression levels of TLR2 ,TLR4 mRNA and TNF-αprotein in group C at the 3rd day was higher than the 1st day(P<0 .01) ,meanwhile ,the expression of TLR4 was higher than in group A(P<0 .05) .Conclusion TNF-αand TLR2 expression maybe involve in the infection and process of candida glabrata ,and TLR4 may play a synergistic effect .
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Objective To analyze the influence of vasoactive drugs on hemodynamics in septic shock patients and to explore the guiding value of systemic vascular resistance index (SVRI) for the application of vasoactive drugs .Methods 21 cases of septic shock in ICU from March 2010 to April 2012 were retrospectively analyzed .The PiCCO monitor technique was applied to monitor the he-modynamic parameters including the cardiac index (CI) ,intrathoracic blood volume index (ITBI) ,systemic cenous resistance index (SVRI) and mean arterial pressure(MAP) ,once per 6-8 h .ITBI>850 mL/m2 was taken as the normal criterion .The data of 202 cases meeting the criterion were collected .These cases were divided into two groups according to whether MAP reach 65 mm Hg , the differences of hemodynamic parameters between the two groups were compared .The MAP≥65 mm Hg group was redivided in-to the group 1 ,2 and 3 according to SVRI .Blood characteristics of pressure ,CI and ITBI were compared among these 3 groups .Re-sults The ITBI value in the MAP<65 mm Hg group and the MAP≥65 mm Hg group was higher than the normal value .The CI and SVRI values in the MAP≥65 mm Hg group were higher than those in the MAP< 65 mm Hg group(P<0 .05) ,while the ITBI value had no difference between the two groups .The ITBI values in the three groups with different SVRI all were higher than the upper normal limit ,furthermore ,in the comparisons among 3 groups ,only comparison between the group 1 and the group 3 had sta-tistical difference(P= 0 .47) .When SVRI was lower ,the CI value was higher than the normal value .Along with the increase of SVRI ,MAP increased gradually ,and CI gradually declined .When SVRI was too high ,MAP had no significant increase ,but the CI value was lower than the lower limit .Conclusion The SVRI monitoring by PiCCO can help to guide the application of vasoactive drugs in septic shock patients ,lower SVRI means the declined vascular reaction and higher SVRI indicates the low cardiac function , moreover blood pressure can not be further enhanced by vascular excessive contraction .