人柯萨奇病毒IgA（CoxV-IgA）elisa试剂盒
 
Objective To explore the role ａnd mechanism of non-receptor tyrosine kinase Tec in endotoxin/lipopolysaccharide (LPS)-induced production of interleukin-8(IL-8) in human alveolar epithelial cell A549. Methods Human alveolar epithelial cell A549 was cultured in RPMI-1640 medium containing 10% fetal bovine serum by volume for routine passage, ａnd the second or third generation cells were taken for subsequent experiments. (1) The cells were taken according to random numbers. In LPS group, cells were cultured routinely for 1 hour, ａnd then stimulated with 1μg/mL LPS for 1 hour. The cells in LFM-A13 group were treated with 75 μmol/L LFM-A13 in conventional culture medium for 1 hour, ａnd then changed the culture medium for conventional culture for 1 hour. 25 μmol/L LFM-A13+ LPS group, 75 μmol/L LFM-A13+LPS group ａnd 100 μmol/L LFM-A13+ LPS group were added with 25, 75 ａnd 100 μ mol/l LFM-A13 in conventional culture medium for 1 hour. All the cells were changed ａnd stimulated with 1 μg/mL LPS for 1 hour. The protein expression of Tec in each group was detected by western blotting, ａnd the content of IL-8 in the supernatant of each group was detected by enzyme-linked immunosorbent assay. (2) The cells were randomly divided into 5 groups with 4 holes in each group. The cells in the blank control group were routinely cultured for 2 hours. The cells of small interfering RNA(siRNA) control +LPS group were transfected with empty lentivirus for 10 h, ａnd then were stimulated by adding 1μg/mL LPS into conventional culture medium for 2 h. Tec mus-298 RNAi+LPS group, Tec mus-299 RNAi+ LPS group ａnd Tec mus-300 RNAi+ LPS group were transfected with lentiviruses carrying tecmus-298 RNAi, tecmus-299 RNAi ａnd tecmus-300 RNAi respectively for 10 hours. 1μg/mL LPS was added into the conventional culture medium to stimulate for 2 hours. the protein expression of Tec in each group was deTected by western blot method, ａnd the Tec-siRNA.(3 with the best effect of silencing tec gene was screened. (3) the cells were divided into 4 groups according to random number table method, with 4 holes in each group. the cells in the blank control group were routinely cultured for 2 hours. The virus cells in the control group were transfected with empty lentivirus for 10 h, ａnd then cultured routinely for 2h; The LPS group cells were stimulated by adding 1μg/mL LPS in the conventional culture medium for 2 hours. Tec-siRNA+ LPS group cells were transfected with lentivirus carrying tec-SiRNA with the best effect of silencing tec gene for 10 h, ａnd then were stimulated by adding 1 μg/mL LPS in the conventional culture medium for 2 h. western blot was used to detect the protein expression of p38 mitogen-activated protein kinase (MAPK) ａnd extracellular signal-regulated kinase (ERK)MAPK in each group. the data were analyzed by one-way ANOVA ａnd LSD-T. 。