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Objective@#To study the correlation between the level of T-bet expression and liver damage in peripheral plasma cells of patients with autoimmune hepatitis (AIH) in order to provide reference for the study of pathogenesis and development of diseases.@*Methods@#The peripheral venous blood and clinical examination data of 29 cases with AIH and 6 healthy volunteers were collected. The percentage of subpopulations of peripheral blood B cells and the proportion of T-bet+ cells in each subgroup were detected by flow cytometry. Plasma cells (CD19+CD10-CD27hiCD38hi), primary B cells (CD19+CD10-CD27-IgD+), transitional B cells (CD19+CD10+), and memory B cells (CD19+CD10-CD27+IgD-) were the included subsets of B cells. Serum immunoglobulin G (IgG) and alanine aminotransferase (ALT) levels, the proportion of B cells in peripheral blood subsets and IgG level, the proportion of T-bet+ cells in each subset and the proportion of T-bet+ plasma cells in each subset in B cells, the proportion of T-bet+ plasma cells and the level of serum ALT were analyzed for correlation analysis. Statistical analysis was performed using two independent sample t-tests and linear regression.@*Results@#The serum IgG level of AIH patients with abnormal ALT (19.47 ± 1.039)g/L was significantly higher than that of normal ALT patients (15.5 ± 1.069)g/L, and the difference was statistically significant (t = 2.65, P < 0.05). The percentage of peripheral plasma cells in B cells of AIH patients (2.80 ± 0.14) % was higher than that of healthy volunteers (0.73 ± 0.09) %, and the difference was statistically significant (P < 0.01). The percentage of T-bet+ cells in peripheral plasma cells of AIH patients (23.54 ± 1.61) % was higher than that of healthy volunteers (6.59±0.59) % , and the difference was statistically significant (P < 0.01). The correlation analysis showed that the proportion of T-bet+ cells in peripheral plasma cells of AIH patients was positively correlated with the proportion of plasma cells to B cells (r = 0.224 7, P < 0.01), and the percentage of peripheral plasma cells to B cells was positively correlated with the level of serum IgG (r = 0.299 1, P < 0.01). Serum IgG level was correlated with the level of ALT, reflecting an indicator of liver damage (t = 2.65, P < 0.05).@*Conclusion@#The increase of T-bet expression in the peripheral plasma cells of AIH patients is associated with liver damage, which is a new mechanism of AIH pathogenesis and disease progression.
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Objective@#To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks.@*Methods@#Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations.@*Results@#Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29).@*Conclusion@#Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.
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Objective The hepatitis B virus (HBV) core protein assembly inhibitors GLS4JHS could destroy HBV capsid assembly and the formation of non-capsid polymer structure.The aim of this study is to explore the mechanisms of GLS4JHS in inhibiting HBV replication.Methods HepAD38 cells was used as the study model.TaqMan real-time polymerase chain reaction (PCR) and quantitative real-time PCR with specific primers were used to measure the change in pregenomic RNA (pgRNA) and covalently closed circular DNA (cccDNA) levels under different concentrations.ChIP assay in HepAD38 cells was used to assess the recruitment of HBV core protein and histone modifications.Results The amount of cccDNA and pgRNA decreased with the increasing GLS4JHS concentrations.After the drug concentrations reached 400 nmol/L, cccDNA and pgRNA declined by 94% and 84%, respectively.Both HBV core protein occupancy on the cccDNA and cccDNA-bound H3 histone acetylation were reduced by GLS4JHS.Conclusions GLS4JHS decreases transcriptional activity of cccDNA and reduces pgRNA production by inhibiting cccDNA minichromosome bound to HBV core protein and acetylated histone H3, which results in HBV DNA formation.
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Objective To investigate the trend of antimicrobial resistance and the prevalence of resistant genes in Pseudomonas aeruginosa strains isolated from Hangzhou First People's Hospital.Methods Antimicrobial susceptibilities of 1489 Pseudomonas aeruginosa strains isolated from 2003 to 2007 were statistically analyzed using WHONET.MICs of 11 antimicrobisis to 36 multi-drug-resistant Pseudomonas aerugionosa strains were determined by agar dilution method.Genes of β-lactamases(BLA)and aminoglycoside-modifying enzymes(AMEs)were detected by PCR and the PCR products were sequenced.Results The resistant rates to aztreonam,imipenem,ceftazidime,cefepime,piperacillin,piperacillin/tazobactam.cefoperazone/sulbactam,ciprofloxacin,levofloxacin,gentamicin and amikacin were increased from 13.4%,10.6%,8.7%,7.9%,12.7%,12.7%,6.7%, 15.8%,20.5%,24.7% and 10.9%in 2003 to 35.3%,40.9%,18.4%, 32.4%,32.9%,32.0%,21.9%,37.8%,38.6%, 39.4% and 34.8% in 2007.respectively.Hish MICs of 11 antimicrobiMs for multi-drug resistant Pseudomonas aerouginosa were determined with MIC90≥128 μg/mL.In 36 multi-drug resistant Pseudomonas aeruginosa strains,21(58.3%)strains carried β-lactamase genes and 32 strains(88.9%)carried aminoglycosidemodifying enzyme genes,while the deletion rate of oprD2 was 80.6%(29/36).Conclusions The resistant rates to common antibiotics of Pseudomonas aeruginosa have increased.resulting in multi-drug resistance.Genes of β-lactamases and aminoglycoside-medifying enzymes are prevalent in multi-drug resistant Pseudomonas aeruginosa strains,with the common deletion of oprD2.
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Objective To identify the spectrum of pathogenic microorganisms of urinary tract infection and the drug resistance in a hospital setting. Methods The pathogenic microorganisms isolated from 3117 mid-stream urine samples of patients admitted in Hangzhou First People's Hospital from 2005 to 2007 and their drug resistance results were retrospectively analyzed. Results Bacteria were the most prevalent microorganisms in the urinary tract infection, and followed by fungus, mycoplasma and chlamydia. Escherichia eoli accounted for the largest proportion of gram-negative bacteria, in which the ESBLs positive strains accounted for 51.2%, and their drug resistance rate was much higher than that of ESBLs negative strains. Main gram-positive coccobacteria was all sensitive to vancomycin, and relatively sensitive to nitrofurantnin and ampicillin. Conclusions Escherichia coli continue to prevail upon the spectrum of pathogenic microorganism of the urinary tract infection, and the fungus, mycoplasma and chlamydia infections are rising. Antibacterial agents should be used under the guidance of drug sensitivity test, and the combined use should be avoidd.
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OBJECTIVE To investigate the resistance and the distribution of the main ?-lactamases encoding gene in Acinetobacter baumannii isolated from four hospitals in Hangzhou city to provide the basic data for the optional treatment of A.baumannii infection.METHODS The identification of A.baumannii was performed using VITEK-AMS60.The minimum inhibitory concentrations(MIC) was examined by agar dilution and E-test.The homology of the resistant isolates was finished by pulsed field gel electrophoresis(PFGE).PCR and sequencing were used to analyze the ?-lactamases encoding gene of the 36 strains of imipenem-resistant A.baumannii.RESULTS All of the imipenem-resistant isolates produced carbapenemase OXA-23,and 5 isolates produced PER-1,2 isolates produced TEM-1 except OXA-23.No metallo-?-lactamases were detected.No plasmid was extracted.Clone transmission of the imipenem-resistant strains existed in the 4 hospitals.Most strains were isolated from intensive care unit(ICU). CONCLUSIONS The clone transmission of imipenem-resistant A.baumannii strains is occurred in 4 hospitals.All strains produce carbapenemase OXA-23.Five strains also produce PER-1 type extended-spectrum ?-lactamases.Two strains also produce TEM-1 type extended-spectrum ?-lactamases.
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OBJECTIVE To investigate the distribution and the antimicrobial resistance of Alcaligenes xylosoxidans and to provide the guidance of rational use of antibiotics.METHODS A.xylosoxidans was isolated and identified through VITEK-AMS all-automatic microbiology analysis system and its G~-bacilli identification cards(GNI) and drug sensitivity cards(GNS-KI/121) were also analyzed.Drug-resistance was tested by Kirby-Bauer disk(sensitivity) method((Bio-Merieux) and Oxoid products).RESULTS The drug-resistance rates to the 1st to 4th((except) CAZ) generation cephalosporins were more than 77%.The drug-resistance rate to the aminoglycosides was more than 76%,and to(ampicillin,) amoxicillin/CA,ampicillin/sulbactam,cefoxitin and cefuroxime were all 100%;but to(aztreonam) was 92.86%;66.67% and 44.44% strains were resistant to ciprofloxacin and(levofloxacin).(CONCLUSIONS) The detection rates of the multidrug resistant A.xylosoxidans have the tendency to increase yearly.The antibiotics should be used reasonably according to the results of drug sensitivity test in clinic.
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OBJECTIVE To develop a real-time fluorescence PCR assay to detect the genes encoding thermolabile(hemolysin)(TLH),thermostable direct hemolysin(TDH) and TDH-related hemolysin(TRH) of Vibrio(parahaemolyticus).METHODS The genes of TDH and TRH were selected as target ones of thermostable direct and TDH-related hemolysin,and TLH gene as a specific genomic marker for V.parahaemolyticus.Designed and synthesized the primers and Taqman probes,we investigated 487 stool samples of doubt foodborne illness patients by real-time fluorescence PCR.RESULTS The sensitivity of the assay for TLH and TDH was 1.0?10~2copies,but the sensitivity of TRH was 1.0?10~3copies. Among 487 samples,112 V.parahaemolyticus strains were found;101 samples of these strains showed the production of TDH;none of them was positive for TRH.CONCLUSIONS The Taqman PCR is a rapid and sensitive method to detect the TLH,TDH and TRH of V.parahaemolyticus,it is well suited for screening large numbers of samples at the same time;and TDH is one of the primary virulence factors in clinical isolated V. parahaemolyticus.
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OBJECTIVE To investigate the distribution of pathogens and their antibiotic resistance in lower respiratory tract infection(LRTI) from intensive care unit(ICU) in our hospital,and provide basis for rational selection of clinical drugs.METHODS Pathogens were detected from qualified sputum specimens in LRTI from ICU and identified by VITEK-AMS60 automatic microbial analyzing system.Drug susceptibility was determined by KB test.RESULTS From 320 sputum specimens 367 pathogens were detected between from Jan 2007 to Mar 2008,including 261 strains(71.1%) of Gram-negative bacilli,70 strains(19.1%) of fungi,and 36 strains(9.8%) of Gram-positive cocci.21.8% Of the isolated pathogens were Acinetobacter baumannii,with 16.7% of drug-resistant rate to cefoperazone/sulbactam and over 71% to other 13 antibiotic agents.The rate of extended spectrum ?-lactamases(ESBLs) producing Escherichia coli isolates and Klebsiella pneumoniae ones were 65.2% and 72.0%,respectively,comparing to 84.6% for meticillin-resistant Staphylococcus aureus(MRSA).CONCLUSIONS Gram-negative bacilli are the major pathogens of LRTI in ICU,in which A.baumannii shows with a high rate of drug-resistance,followed by fungi,which should attract the clinician′s more attention.
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Objectives To investigate the properties and distributions of ampC gene among different drug-resistant strains of Serratia marcescens,and the relationship of control gene ampR with AmpC enzymes′ expressions.Methods According to the results of inducting experiment with 1/2 MIC of beta-lactam antibiotics (CTX),three-dimensional testing and isoelectric focusing electrophoresis testing,143 strains of S.marcescens were classified into three groups:including induction group, continuous low-production group and hyperproduction group. In each group, the sequences of ampC and ampR genes were amplified using the method of PCR. The products of PCR were analyzed. The plasmid-mediated beta-lactamases were detected using the method of conjugation experiment.Results Among 143 strains of S.marcescens, the continuous low -production strains, induction strains and hyperproduction strains were 14,103,and 18, respectively.125 and 99 strains were ampC and ampR gene positive, respectively.The detection rate of ampR in hyperproduction group was lower than other groups.5 sites of ampC genes and 4 sites in the Open Reading Frame (ORF) of ampR gene were easily mutated in 5 induction strains and 2 hyperproduction strains.Conclusions The production of inducing drug-resistance of some S.marcescens might be related to mutation of ampC gene encoding AmpC beta-lactamases and the ORF mutation in ampR. The continuous hyperproduction drug-resistance had something to do with deletion mutation in ampR in segmental hyperproduction strains.The plasmid-mediated AmpC enzymes hadn′t been found in S.marcescens.