Objective:To investigate the effect of bone morphogenetic protein-9(BMP9)overexpression on inflammatory response and apoptosis of alveolar epithelial cells induced by lipopolysaccharide(LPS).Methods:A549 cells were stimulated with 0.1 μg/ml LPS,expressions and changes of BMP9 protein at different time points(LPS stimulation for 0 h,6 h,12 h,24 h)were detected by Western blot.Expression of BMP9 in A549 cells was up-regulated by transfection of BMP9 plasmid,and the transfection efficiency was verified by Western blot and qPCR.After 12 h of LPS stimulation,levels of inflammatory factors TNF-α,IL-6,IL-1β in cell supernatant were detected by ELISA,expressions of anti-apoptotic protein(Bcl-2)and pro-apoptotic protein(Bax)were detected by Western blot,and apoptosis of cells was detected by TUNEL staining.Results:With the extension of LPS stimulation time,expression of BMP9 was down-regulated.Overexpression of BMP9 successfully up-regulated expression of BMP9 in A549 cells.LPS stimulation promoted secretions of TNF-α,IL-6 and IL-1β from A549 cells,increased apoptosis,promoted Bcl-2 expression while inhibited Bax expression.Overexpression of BMP9 inhibited TNF-α,IL-6 and IL-1β releasing,decreased apoptosis,inhibited Bcl-2 expression,while promoted Bax expression.Conclusion:In LPS-stimulated A549 cells,BMP9 expression is gradually decreased at a time-depen-dent manner.Overexpression of BMP9 can inhibit LPS-induced inflammatory response and apoptosis in A549 cells.

Objective:To investigate the effect of bone morphogenetic protein-9(BMP9)overexpression on inflammatory response and apoptosis of alveolar epithelial cells induced by lipopolysaccharide(LPS).Methods:A549 cells were stimulated with 0.1 μg/ml LPS,expressions and changes of BMP9 protein at different time points(LPS stimulation for 0 h,6 h,12 h,24 h)were detected by Western blot.Expression of BMP9 in A549 cells was up-regulated by transfection of BMP9 plasmid,and the transfection efficiency was verified by Western blot and qPCR.After 12 h of LPS stimulation,levels of inflammatory factors TNF-α,IL-6,IL-1β in cell supernatant were detected by ELISA,expressions of anti-apoptotic protein(Bcl-2)and pro-apoptotic protein(Bax)were detected by Western blot,and apoptosis of cells was detected by TUNEL staining.Results:With the extension of LPS stimulation time,expression of BMP9 was down-regulated.Overexpression of BMP9 successfully up-regulated expression of BMP9 in A549 cells.LPS stimulation promoted secretions of TNF-α,IL-6 and IL-1β from A549 cells,increased apoptosis,promoted Bcl-2 expression while inhibited Bax expression.Overexpression of BMP9 inhibited TNF-α,IL-6 and IL-1β releasing,decreased apoptosis,inhibited Bcl-2 expression,while promoted Bax expression.Conclusion:In LPS-stimulated A549 cells,BMP9 expression is gradually decreased at a time-depen-dent manner.Overexpression of BMP9 can inhibit LPS-induced inflammatory response and apoptosis in A549 cells.

ObjectiveTo investigate the clinical significance of highly sensitive nucleic acid detection in precise antiviral therapy for patients with liver cirrhosis and its association with aminotransferase level. MethodsA total of 377 patients with hepatitis B cirrhosis who were hospitalized or attended the outpatient service from May 2013 to April 2019 were enrolled and tested by both domestic HBV DNA detection and highly sensitive Cobas HBV DNA detection. All patients underwent biochemical examination, four blood coagulation tests, routine blood test, and upper abdominal computed tomography or ultrasound. Sensitivity of different HBV DNA detection reagents was compared in liver cirrhosis patients with a low viral load, and the correlation between alanine aminotransferase (ALT) level and viral load was analyzed. The paired t-test was used for comparison of continuous data before and after treatment. The receiver operating characteristic (ROC) curve was used to screen out the optimal predictive values of ALT at different cut-off values of HBV DNA. ResultsAmong the 377 patients with hepatitis B cirrhosis, 215 tested positive and 162 tested negative by domestic HBV DNA, and among these 162 patients, 104 (64.2%) tested positive by Cobas HBV DNA detection, with a mean level of 267.5±42.3 IU/ml. After 24 weeks of antiviral therapy, the 104 patients with hepatitis B cirrhosis had significant improvements in viral replication level, ALT, and Child-Pugh score for liver function; HBV DNA decreased from 267.5±32.2 IU/ml before treatment to 59.6±7.7 IU/ml after treatment (t=3.486, P=0.002), ALT decreased from 871±10.8 U/L before treatment to 36.5±7.6 U/L after treatment (t=3.235, P=0.020), and the Child-Pugh score decreased from 6.5±0.7 before treatment to 5.7±0.5 after treatment (t=2.928, P=0.041). The ROC curve analysis of ALT in predicting HBV DNA decision point showed that an ALT level of 29 IU/L was the most sensitive cut-off value for predicting HBV DNA ＜20 IU/ml, with an area under the ROC curve of 0.904, a sensitivity of 1.0, and a specificity of 0.237. ConclusionPrecise detection helps to guarantee the precise clinical treatment of patients with hepatitis B cirrhosis and improve their treatment outcome and prognosis. An ALT level of 29 IU/L is a sensitive indicator for predicting patients with negative Cobas HBV DNA, so as to achieve individualized precise screening and treatment.

ObjectiveTo investigate the clinical features of patients with failure or recurrence after treatment with PEG-IFN combined with ribavirin (PR regimen) in the real world and the clinical effect of different direct-acting antiviral agent (DAA) regimens in such patients. MethodsA retrospective analysis was performed for the clinical data of 106 patients with chronic hepatitis C or hepatitis C-related compensated liver cirrhosis who attended the outpatient service or were hospitalized in The First Peoples’ s Hospital of Lanzhou from March 2014 to January 2018, and these patients experienced failure or recurrence after the treatment with the standard PR regimen. There were 54 male and 52 female patients. According to the response to PR treatment, the patients were divided into failure group with 13 patients, recurrence group with 51 patients, and sustained virologic response group with 42 patients. All patients underwent IL-28B rs12979860/rs8099917 detection, baseline biochemical examination, Cobas HCV RNA test, and viral genotyping, and these results were compared between groups. The clinical outcomes of patients with failure or recurrence after PR treatment were observed after the treatment with different DAA regimens. The chi-square test was used for comparison of categorical data between groups; a one-way analysis of variance was used for comparison between multiple groups. ResultsThe failure group and the recurrence group had a significantly higher age than the sustained virologic response group (F=14.05, P＜0.001). Among the patients in the failure group, 86.4% had viral genotype 1b, while among those in the recurrence group, 72.5% had viral genotype 2a, and there was a significant difference between the three groups (χ2=17269, P=0.002). Among the patients in the failure group, 92.3% had a baseline HCV RNA level of ≥106 IU/L, and the failure group had a significantly higher proportion of such patients than the recurrence group and the sustained virologic response group (χ2=10.407, P=0.005). There were no significant differences in sex and liver cirrhosis between the three groups (all P＞0.05). Among the patients with primary treatment failure, 100% patients had the non-protective genotype of IL-28B rs12979860 CT/TT, and 92.3% had the non-protective genotype of IL-28B rs8099917 TG/GG; among the patients with recurrence, 84.3% patients had the non-protective genotype of IL-28B rs12979860 CT/TT, and 86.3% had the non-protective genotype of IL-28B rs8099917 TG/GG; among the patients in the sustained virologic response group, 85.7% gad genotype CC at IL-28B rs12979860 and 88.1% had genotype TT at IL-28B rs8099917. There were significant differences in the constituent ratios of rs12979860 and rs8099917 gene polymorphisms between the three groups (χ2=57.263 and 59.651, both P＜0.001). The patients with failure or recurrence after PR treatment achieved a sustained virologic response rate of 100% after the treatment with three different DAA regimens based on sofosbuvir. ConclusionViral genotype and non-protective genotypes at IL-28B rs12979860 and rs8099917 are influencing factors for failure and recurrence after PR treatment. The three different DAA regimens based on sofosbuvir achieves a sustained virologic response rate of 100% and has good safety in patients with failure or recurrence after PR treatment, which is not affected by the factors including IL-28B single nucleotide polymorphism and viral replication level in the host.