Objective To investigate the clinical significance of detection of hepatitis E virus (HEV) RNA in sera from patients with acute hepatitis E using real-time reverse transcription (RT)-PCR to detect hepatitis E virus RNA in sera from patients with acute hepatitis E.Methods A real-time RT-PCR assay, which can amplifies and detect the conserved region on ORF3, was used in this study. 434 outpatients and hospitalized patients with acute HEV infection was enrolled into this study.Simultaneously,the serum samples from 40 patients with HAV infection, 100 patients with HBV infection and 110 healthy blood donors were collected as the control The real-time RT-PCR was performed to detect HEV RNA in all these sera.Results 232 sera (53.5%) were positive for HEV RNA by real-time RT-PCR and all of the control were negative.The results of real-time RT-PCR and anti-HEV IgM (ELISA) were concordant in 67.1% samples.There was significant difference between the two methods ( Kappa = 0.308, P = 0.000 ).The first serum sample from five serum samples of the patients was positive for HEV RNA and negative for anti-HEV IgM.Follow-up studies showed all the five sera samples were positive for anti-HEV IgM.HEV RNA in serum could be detected between 2 and 10 days.Conclusions The real-time fluorescent RT-PCR method has high specificity, and can be applied to the qualitative detection of the serum with genotypes Ⅰ and Ⅳ of hepatitis E virus.Its clinical use can improve the early diagnosis of HEV.

s E in Beijing belong to HEV genotype Ⅳ.

Objective To investigate the related factors and pathogens of ventilator-associated pneumonia (VAP) after heart surgery.Methods VAP was diagnosed in 105 patients out of 1688 cases (6.2％) who underwent heart surgery in our department between January 2004 and January 2011.Clinical data,pathogens and treatments were analyzed.Results Incidence of VAP was 6.2％ (105/1688),and 53.0％ (105/198) in patients who required more than 48 hours of mechanical ventilation.One hundred and ninety-eight pathogen strains were isolated by bacterial culture,in which Gram negative bacteria accounted for 69.2％ (137/198),Gram positive bacteria 27.8 ％ (55/198),and fungi for 3.0％ (6/198).Pseudomonas aeruginosa,klebsiella pneumoniae,acinetobacter baumannii,escherichia coli,and staphylocccus aureus were the main pathogens of VAP.The independent risk factors for VAP were:age ＞ 70 years,emergent surgery,perioperative transfusions,reintubation and days of mechanical ventilation(all P ＜ 0.01).Median length of stay in the ICU for patients who developed VAP or not was (24.7 ± 4.5) days versus (3.2 ±1.5)days,respectively (P ＜0.05) and in-hospital mortality was 25.7％ (27/105) versus 2.9％(46/1583) respectively (P ＜ 0.05).Conclusions Patients undergoing heart surgery have a high frequency of developing VAP,especially in patients that require more than 48 hours of mechanical ventilation.VAP is associated with high in-hospital mortality.Age ＞ 70 years,emergent surgery,perioperative transfusions,reintubation and prolonged mechanical ventilation use are independent risk factors for VAP in patients following cardiac surgery.Pseudomouas aeruginosa,klebsiella pneumoniae,acinetobacter baumannii,escherichia coli,and staphylocccus aureus are the main pathogens of VAP.

Objective:To analyze the genes related to platelet activation in essential thrombocythemia (ET)based on transcriptome sequencing technology (RNA-seq ),and to explore the potential targets related to ET thrombosis. Methods:Blood samples from ET patients and healthy individuals were collected for RNA-seq,and differentially expressed lncRNAs,miRNAs,and mRNAs were selected to construct a lncRNA-miRNA-mRNA regulatory network. Differential mRNAs in the regulatory network were enriched and analyzed using Gene Ontology (GO ) and Kyoto Encyclopedia of Genes and Genomes (KEGG).The real-time PCR method was applied to validate differential mRNAs on crucial signaling pathways.Results:A total of 32 lncRNAs (3 up-regulated,29 down-regulated),16 miRNAs (8 up-regulated,8 down-regulated),and 35 mRNAs (27 up-regulated,8 down-regulated)were identified as differentially expressed.Among them,5 lncRNAs,12 miRNAs,and 19 mRNAs constituted the regulatory network.KEGG enrichment analysis showed that the differential mRNAs were related to the platelet activation signaling pathway,and there were 6 differential mRNAs related to platelet activation,namely F2R,ITGA2B,ITGB1,ITGB3,PTGS1,and GP1 BB,which were all up-regulated in their expression.RT-PCR results showed that the expression of five mRNAs including F2R,ITGA2B,ITGB1,ITGB3,and GP1BB were upregulated in ET patients compared with healthy subjects,and consistent with RNA-seq results,while PTGS1 expression was not significantly different.Conclusion:Differential mRNAs in ET patients are related to the platelet activation pathway,and F2R,ITGA2B,ITGB1,ITGB3,and GP1BB mRNAs may serve as novel targets associated with platelet activation in ET.