Objective:To determine the spectrum drift characteristics of CI4+CD25+Tregs TCR β chain CDR3 in patients with different phases of acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients before and after the entecavir treatment.Methods:Anticoagulation venous blood was collected from 4 normal control subjects,3 AHB patients with acute phase and convalescent phase,and 4 CHB patients before and after the entecavir treatment;and peripheral blood mononuclear cells were isolated;CD4+ CD25+ Tregs were separated by using the magnetic beads,and total RNAs were extracted from CD4+ CD25+ Tregs and used for reverse transcription.The TRBV CDR3 was amplified by polymerase chain reaction (PCR) with forward primers specific for 24 TRBV families and one fluorescence-labeled common reverse primer specific for the BC region.The PCR products were sent out for Genescan,and results were analyzed for the TRBV family CDR3 spectrum characteristics by using the Peak Scanner Software vl.0.Data were analyzed with the comparative t-test to perform the statistical analysis.Results:The CDR3 spectral types of the TRBV family showed drift characteristics in 3 cases of AHB patients with acute and convalescent phases;single/oligo peak spectral type family was observed in most of patients with acute phase;multiple peak spectral type was seen in patients with convalescent phase;and the common spectrum shift of TRBV4,10,14,16,19 families seen in patients with acute phase was changed to multiple peak spectral type.The clonal expansion of TRBV family in the CD4+CD25+Tregs in PBMC from AHB patients with convalescent phase was significantly lower than AHB patients with acute phase (t =9.456,P =0.011).The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in C HB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.The clonal expansion of TRBV family in the CI4+CD25+Tregs in PBMC from of CHB patients after antiviral treatment was increased (t =-0.666,P =0.553).Conclusion:TRBV4,10,14,16,19 family of spectrum shift seen in AHB patients with acute phase was changed to multiple peak spectral type in patients with convalescent phase,suggesting this transition may be associated with HBsAg and HBeAg turning to negative.The clonal expansion of Tregs TRBV13.2,15,16,18,20 family seen in CHB patients before treatment may interfere the virus removal through down-regulating the body's immune response;and with the decline of viral load in serum after the antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24 family may help body induce immune tolerance and result in the HBV persistence.

Objective To analyse the spectral patterns of complementarity determining region 3 (CDR3) length distribution of T lymphocyte receptor beta chain variable (TRBV) gene families in infiltrating T cells of the liver tissues and the peripheral blood samples of patients with chronic hepatitis B (CHB) in order to evaluate the characteristics of T cell clonal expansion. Methods The spectral patterns drift of TRBV gene families (the monoclonal/oligoclonal TCR β T cells) in the peripheral blood and hepatic tissues from 11 cases of CHB patients were analyzed by the real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-PCR) with DNA melting curve analysis, and abnormal rates of TRBV gene families were compared between CHB patients and healthy control. The comparison of rates was done by chi square test. Results The gene melting spectral pattern of 26 TRBV families of the 11 CHB patients, no matter in the peripheral blood or hepatic tissue, showed either a single peak or prominent melting peaks, even disappeared for certain TRBV families. The abnormal rate of TRBV gene families in the hepatic tissues was significantly higher than that in the peripheral blood samples (x2 = 23. 246, P＜0. 01). What is more interesting was that some parts of TRBV families were identical in both the peripheral blood and the hepatic tissue in certain patients. TCR BV13.1, TCR BV17 and TCR BV22 fragments were found to be restricted used in both the peripheral blood and hepatic tissue by some CHB patients. Conclusions T cells in the peripheral blood and the hepatic tissues of CHB patients can develop clonal expansion to some extent.Parts of TRBV families are restricted used in the peripheral blood and hepatic tissue in some CHB patients, which offers a foundation for further studying the common specific spectral drift patterns of TRBV CDR3 gene in CHB patients.

Objective To investigate the common bacteria and change in antimicrobial resistance in an intensive care unit (ICU)in the past 5 years,and provide evidence for rational use of antimicrobial agents.Methods Bacteria isolated from ICU patients in a tertiary first-class hospital from 2009 to 2013 were collected,identified,and per-formed antimicrobial susceptibility testing.Results A total of 1 196 bacteria isolates were isolated in 2009-2013, the top five species were Acinetobacter baumannii (A.baumannii,29.60%),Pseudomonas aeruginosa (P . aeruginosa,14.38%),Staphylococcus aureus (S .aureus ,12.21 %),Escherichia coli (E.coli,12.21 %),and Klebsiella pneumoniae (K .pneumoniae ,11 .37%).Resistance rates of S .aureus to oxacillin,gentamycin, clindamycim,ciprofloxacin,and rifampicin showed a decreasing tendency from 2009 to 2013(all P <0.05),and no strain was detected to be resistant to vancomycin during 5 years.Resistance rates of P .aeruginosa to ceftazidime, cefepime,aztreonam,gentamycin,amikacin,tobramycin,and piperacillin/tazobactam decreased gradually (all P <0.05),imipenem resistance rates were 32.26% -46.43% in 2009 -2012 and 16.00% in 2013;A.baumannii maintained a low level resistance to amikacin,tobramycin,and compound sulfamethoxazole(all P <0.05),resistance rates to imipenem were all >80% in 2009-2011 and 10.53% in 2013,A.baumannii had high resistance rates to most antimicrobial agents(resistance rates were >80%)during 5 years.Resistance rates of K .pneumoniae and E. coli to piperacillin/tazobactam,cefazolin,cefepime,amikacin,and aztreonam had a decreased tendency in 2009 -2013(all P <0.05).Conclusion The common bacteria causing infection in the ICU of this hospital showed a down-ward trend,which may be related to the introduction of national policies and management of hospital,continuous management of antimicrobial agents is suggested,antimicrobial agents should be used rationally to prevent the in-crease of bacterial resistance.

Objective To analyze risk factors and complication characteristics of healthcare-associated infection (HAI)in patients with lung cancer,and provide evidence for the formulation of HAI management strategy. Methods HAI-related articles were retrieved from China Biology Medicine (CBM),China National Knowledge Infrastructure (CNKI),Wanfang database,Vip database,PubMed,and Embase,all data were conducted Meta-analysis.Results A total of 19 articles involving 8 069 hospitalized patients with lung cancer (1 280 had HAI)were included.Meta-analysis on combined values of medical factors for HAI were as follows:OR(95%CI )of anti-tumor therapy(radiotherapy and chemotherapy),number of chemotherapy (≥ 2 times ),antimicrobial prophylaxis, immunosuppressant therapy,and invasive operation were 3.13 (1 .82,5.39),9.20 (3.04,27.87),3.23 (1 .77, 5.91),2.00(1 .56,2.57),and 2.28(1 .81 ,2.88),respectively;Meta-analysis on combined values of complication factors for HAI were as follows:OR (95% CI )of pulmonary diseases,chronic obstructive pulmonary disease (COPD),diabetes,renal dysfunction,malnutrition,hypoalbuminemia,neutropenia,and leukopenia were 2.65 (1 .74,4.02),2.40 (1 .76,3.27),2.25 (1 .85,2.73 ),2.56 (1 .18,5.52),5.51 (1 .70,17.89),2.05 (1 .56, 2.70),3.38(1 .40,8.18),and 2.10 (1 .22,3.62),respectively.Conclusion HAI-related factors of medical treat-ment and complications in patients with lung cancer are diversity,risk factors for HAI in patients with lung cancer are anti-tumor therapy,immunosuppressant therapy,antimicrobial prophylaxis,invasive operation,pulmonary dis-eases,COPD,diabetes,renal dysfunction,malnutrition,hypoalbuminemia,neutropenia,and leucopenia.

Objective To provide the evidences for the management strategies of ventilator-associated pneumonia (VAP) in neonates,we systematically reviewed all related studies and analyzed the high-risk primary disease and medical factors of VAP in neonates.Methods We retrieved all related studies in CNKI,Wanfang,VIP,CBM,Pubmed and Embase and evaluated their quality by Newcastle-Ottawa Scale and analyzed all data by qualitative and Meta-analysis.Results There were 12 case-control studies with higher methodological quality and involving 1 994 neonates and with 708 VAP patients.Six studies involving 872 neonates were included,the odds ratio of respiratory distress syndrome(OR=2.81) and malnutrition(OR=5.18) had significant differences between VAP and non-VAP group.Seven studies involving 1 110 neonates were included and the odds ratio of patients with corticosteroids (OR=3.12),central inhibitors (OR=2.31),antacids (OR=4.35) and Gamma globulin with large doses (OR=2.35) had significant differences between VAP group and non VAP.Four studies involving 554 neonates were included and the odds ratio of patients with closed chest drainage (OR=1.81)and umbilical vein catheterization (OR=9.19) had significant differences between VAP group and non VAP.Six studies involving 1 139 neonates were included and the odds ratio of patients with parenteral nutrition (OR=1.82)and blood transfusions (OR=2.49) had significant differences between VAP group and non VAP.Conclusions Our study confirms that the respiratory distress syndrome and malnutrition corticosteroids,central inhibitors,antacids,Gamma globulin with large doses,closed chest drainage,umbilical vein catheterization,parenteral nutrition and blood transfusions are important risk and early-warning factors.

CD4-Positive T-Lymphocytes ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Interleukin-2 Receptor alpha Subunit ; Sequence Analysis

OBJECTIVETo investigate the role ofCD4+CD25+ T regulatory (Treg) cells, T helper (Th)17cells and interleukin (IL)-6 in the progression of hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) and determine their value as prognostic markers.

METHODSThe Chinese National Knowledge Infrastructure (CNKI), WanFang, Chinese Scientific Journals (VIP), PubMed, Embase and Web of Science databases were searched for English language case-control studies on the relationship between regulatory T lymphocytes and ACLF.The quality of included studies was assessed using the Newcastle-Ottawa scale. The meta-analysis was designed according to the PICOS approach recommended by the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement. RevMan software, version 5.1, was used to perform the meta-analysis.

RESULTSNine case-cohort studies were selected for inclusion in the metaanalysis.The results of the meta-analyses showed that the level of CD4+CD25+ Treg cells was not significantly different between patients with HBV-related ACLF and patients with chronic hepatitis B (CHB) (mean difference (MD)=0.59, 95% confidence interval (CI)-1.68, 2.85, P=0.61) nor between patients with HBVrelated ACLF and healthy controls (MD=1.12, 95% CI:-1.42, 3.66, P=0.39). Thus, it appears that ACLF patients do not have a higher level of CD4+CD25+ Treg cells than CHB patients or healthy controls. However, the ACLF patients did appear to have a significantly higher level of Th17 cells than both the CHB patients (MD=1.73, 95% CI:0.21, 3.26, P=0.03) and the healthy controls (MD=1.62, 95% CI:(0.52, 2.72, P=0.004). In addition, the ACLF patients also had significantly higher level than both the CHB patients (MD=11.69, 95%CI:1.98, 21.40, P=0.02) and the healthy controls (MD=13.17, 95% CI:1.38, 24.95, P=0.03).

CONCLUSIONCD4+CD25+ Treg cells may be an important protective factor in the progression and prognosis of HBV-related ACLF, while Thl7 cells and IL-6 may be risk factors for further progression and worsened prognosis.

Acute-On-Chronic Liver Failure ; diagnosis ; immunology ; CD8-Positive T-Lymphocytes ; Case-Control Studies ; Disease Progression ; Hepatitis B virus ; Hepatitis B, Chronic ; complications ; Humans ; Interleukin-6 ; immunology ; Prognosis ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology