Objective:To investigate the effect of protein tyrosine kinase 6(PTK6) on TNF-α induced human airway epithelial barrier dysfunction and mechamism.Methods: After cultivating 16HBE cells in vitro,the recombined PTK6 and PTK6 siRNA was respectively transfected into the cells,with empty vector and scramble siRNA as control.The cells were incubated with exogenous TNF-α.Cell viability was detected by MTT assay.Cells TER and permeability were detected.ZO-1 and Occludin mRNA were analyzed by RT-PCR.PTK6,ZO-1,Occludin,p-ERK1/2,p-JNK1/2 and p-p38MAPK protein were assayed by Western blot.Results: TNF-α remarkably decreased ZO-1 and Occludin mRNA and protein and TER;increased cells permeability,as well as p-ERK1/2,p-JNK1/2 and p-p38 protein(P<0.05).Upregulation of PTK6 further decreased ZO-1 and Occludin mRNA and protein and TER,enhanced cells permeability and p-JNK1/2 and p-p38 protein(P<0.05),and downregulated PTK6 ,the changes of these indices were opposite(P<0.05).Whereas upregulation or downregulation of PTK6 had no effect on p-ERK1/2.Conclusion: Downregulation of PTK6 inhibits the phosphorylation of JNK1/2 and p38MAPK,thus improving TNF-α-induced human airway epithelial barrier dysfunction.

Objective: To explore the effect of caveolin-1 ( Cav-1 ) on lipopolysaccharide ( LPS )-induced airway mucous hypersecretion.Methods:16HBE human airway epithelial cells with Toll-like receptor 4(TLR4) inhibitor,nuclear factor-kappa B(NF-κB) inhibitor,Cav-1 siRNA or plasmid pr-treated,further stimulated with LPS.The cells were divided into 8 groups:the control group, the LPS group,the LPS+Cav-1 expression group,the LPS+Cav-1 siRNA group,the LPS +negative siRNA group,the LPS +empty vector group,the LPS +E5564 group, the LPS +PDTC group.Cell survival rate was detected by MTT assay.Transcription level of mucin(MUC)5AC was evaluated with RT-PCR.The level of MUC5AC protein was measured by ELISA.The expression of TLR4,Cav-1 and phosphorylated IκBα( p-IκBα) were measured by Western blot.MUC5AC protein changes were observed by immunofluorescence and confocal laser technology.Results:LPS remarkably increased MUC5AC,as well as TLR4,p-IκBα(P<0.05).These effects were prevented by E5564 and PDTC.We found that the overexpression of Cav-1 further enhanced the expression of TLR4, p-IκBαand MUC5AC.However,downregulation of Cav-1 inhibited the expression of TLR4,p-IκBα,MUC5AC.Conclusion: Cav-1 enhances LPS-induced MUC5AC hypersecretion through TLR4/NF-κB signaling pathway.

Objective To explore the effects of Xingnaojing injection on plasma MMP-9 and TIMP-1 levels for patients with cerebral hemorrhage. Methods A total of 60 patients with acute cerebral hemorrhage from February 2014 to May 2015 were selected as research subjects and randomly divided into the research group and the control group, with 30 patients in each group. The research group was given Xingnaojing injection on the basis of regular treatment, and the control group was given regular treatment. The plasma MMP-9 and TIMP-1 levels of the two groups of patients were compared, and the correlation between MMP-9 and TIMP-1 levels in the patients and their correlation with cere-bral edema were analyzed. Results On the fifth day of onset of the disease in the two groups of patients, MMP-9 and TIMP-1 levels were significantly increased, and the levels were reduced on the 14th day. On the fifth day and 14th day of onset of the disease in the research group, MMP-9 level was significantly lower than that in the control group, and TIMP-1 level was significantly higher than that in the control group. The differences were statistically significant (P<0.01). In 24 h, the volume of cerebral edema in the two groups was positively correlated to MMP-9 (r=0.682, P=0.761), and was significantly negatively correlated to TIMP-1(r=-0.489, P=-0.619); on the 14th day, the volume of cerebral edema was positively correlated to MMP-9 (r=0.516, P=0.835). Conclusion Xingnaojing is able to significantly reduce the increasing degree of MMP-1 in the patients with cerebral hemorrhage, enhance the increasing degree of TIMP-1, and improve patients' cerebral edema.