Objective To establish a real -time quantitative PCR ( RT-PCR) assay for detecting serum miR -122, miR-22, and evaluate the clinical significance of miR -122 and miR-22 in patients with chronic hepatitis B ( CHB) by using of this assay .Meth-ods The mature miRNAs were reversely transcripted by using of stem -loop primers .SYBR GreenⅠquantitative real-time PCR ( qRT-PCR) was used for quantification of the miRNAs .The sensitivity of this assay was evaluated by using of the 10-fold-diluted miRNA-122 cDNA standards and the specificity was verified by using of melting curve assay .The accuracy was assessed by intra -assay coeffi-cient of variation (CV) of threshold cycle (Ct value), which were calculated from a 20-times-repeat detection of the miR -122 cDNA (2 ×105 , 2 ×106 , 2 ×107 copies/μl) standards.Using the established qRT -PCR assay, we detected the expression of serum miR -122 and miR-22 in the patients with CHB and healthy controls .Results The qRT-PCR assay exhibited good performances in the linear range, sensitivity and reproducibility while detecting miR -122 and miR-22.The relative level of miR -122 and miR-22 was 17.88 vs 5.35 in the CHB patients and 1.80 vs 1.67 in the controls (P=0.000).Conclusion Using of stem-loop primers, we established a qRT-PCR assay for detection of serum miR -122 and miR-22.Serum miR-122 and miR-22 increased significantly in the CHB pa-tients.

OBJECTIVE To analyze the pathogenic distribution and antimicrobial resistance in our hospital in 2006 and provide the rational information to use antibiotics reasonably.METHODS Flora cultivation and isolation were operated with the methods described by the National Clinical Laboratory Operational Regulations.Flora was identified with the VITEK32 automatic identifier,and bacteria-susceptibility test was operated with Kirby-Bauer method.RESULTS Totally 967 strains of pathogenic bacteria were isolated;they comprised 326 strains of Gram-positive bacteria,541 strains of Gram-negative bacteria and 100 strains of fungi.The main Gram-positive microorganisms included Staphylococcus aureus and Enterococcus faecalis,et al.The main Gram-negative microorganisms included Escherichia coli,Klebsiella pneumoniae,Acinetobacter baumannii,Stenotrophomonas maltophilia and Pseudomonas aerugiinosa,et al.Specimen samples mainly isolated from sputum(43.85%),urine(22.34%),and secretion(10.03%).G+ microorganisms were sensitive to nitrofurantoin and vancomycin.G-microorganisms except A.baumannii and S.maltophilia were sensitive to cefoxitin,piperacillin/tazobactam,imipenem,and amikacin;the average resistant rates of A.baumannii and S.maltophilia to antibiotics were 68.20% and 64.43%,respectively.CONCLUSIONS The severe degree of bacterial multi-drug resistance is increasing,it is urgent to carry out surveillance of bacterial resistance for reasonabe use of antibiotics and decreasing the morbidity rate and the fatality rate.

Objective:To investigate whether SKA1 is a key molecule regulating malignant proliferation of liver cancer, and further explore its mechanism to provide molecular theoretical basis for subsequent targeted therapy.Methods:The data of liver cancer from TCGA database were analyzed by bioinformatics technology. The expression of SKA1 in liver cancer was analyzed. At the same time, we also analyzed the relationship between the expression of SKA1 and the prognosis of patients with liver cancer. The hepatoma cell line overexpressing SKA1 was constructed by liposome-mediated cell transfection technique, and the effect of SKA1 on the proliferation of hepatoma cells was further tested by CCK-8 and plate cloning assay. At the same time, we found that E2F1 is also highly expressed in liver cancer, using bioinformatics technology to analyze the correlation between SKA1 and E2F1 expression, further detecting the binding site of E2F1 in the SKA1 promoter region, and using dual luciferase technology to detect E2F1 against SKA1. Transcriptional activation.Results:KA1 was highly expressed in liver cancer tissues, and the overall survival rate of liver cancer patients with high SKA1 expression was 49.8%, lower than that of patients with low SKA1 expression, showing a negative correlation. E2F1 is also highly expressed in liver cancer tissues, and the survival time of patients with liver cancer with high E2F1 expression is significantly lower than that in the low expression group, which was negatively correlated with poor prognosis. SKA1 overexpression could increase the proliferation ability of liver cancer cells by nearly 50%. SKA1 is regulated by the E2F1 transcription factor, and the E2F1 transcription factor is combined with the SKA1 promoter to transcriptionally activate the expression of SKA1 in liver cancer cells.Conclusion:E2F1 transcriptional activation of SKA1 promotes proliferation of hepatoma cells, leading to poor prognosis in patients with liver cancer