Objective To investigate the balance of Th1/Th2 cytokines and its relationship with prognosis of severe chronic hepatitis B ( CHB ). Methods Peripheral blood samples were collected from 112 severe CHB patients, 30 CHB patients and 30 healthy controls. IL-4, IFN-γ levels and HBV DNA loads were measured by ELISA and fluorescent PCR, respectively. The levels of cytokines in different stages, and their correlations with HBV DNA loads and short-term prognosis were analyzed. Results Higher levels of IL-4, IFN-γ and Th1/Th2 ratios in peripheral blood were detected in patients with severe CHB than those with CHB and the healthy controls (Z = 8.968, 10. 004 and 26. 067, P =0. 009, 0. 007 and 0. 000). IL4 levels in patients with end-stage server CHB were markedly higher than those in other stages ( Z = 3. 672 and 3. 158, P= 0.000 and 0.002), while their Thl/Th2 ratios were lower (Z=3. 161 and 2. 166, P=0.002 and 0. 030). No significant differences on levels of IL-4, IFN-γ and Th1/Th2 ratios were observed in severe CHB patients with different HBV DNA levels (Z =4.431, 2.626 and 0. 140, P =0.219, 0.403 and 0. 987). Elevated IL-4 was closely correlated with the high case-fatality rate within 12 weeks. Conclusions The balance between Th1 and Th2 cytokines is- disturbed in patients with severe CHB. Thl/Th2 ratio decreases with the aggravation of diseases, which may indicate unfavorable short-term prognosis.

Objective To identify the sequence of hepatitis B virus S gene"a"determinant in patients with positive HBsAg and anti-HBs.Methods Nested-PCR Was used to amplify the HBV S gene in 4 patients with positive HBsAg and anti-HBs,and the PCR products were sequenced directly or after cloning.The sequences of"a" determinants were then analyzed by sequence alignment.Results Direct sequencing of PCR products showed that there was one amino acid (aa)residue in"a"determinant less conserved region emerging polymorphism in all 4 patients.Clone sequencing showed that aa residue at 126 of "a"determinant in patient 1 miSht be Thr,Ile and Set,at 134 might be Phe and Set;the aa at 126 in patient 2 misht be Ala and Thr.and in patient 3 might be Ile and Asn;aa polymorphism was not found in patient 4.Conclusion The polymorphism of"a"determinant in HBV S gene might be associated with positivity of both HBsAg and anti-HBs in hepatitis B patients.

Objective To compare the drug resistance of mucoid Pseudomonas aeFagillosa with that of non-mucoid.Methods All specimens isolated and cultured from patients were identified and the antibiotic sensitivity was tested by automatic microorganism analyzer VITEK-32,GNI+,GNS-448,E-teat and K-B.Results Drug resistant rates of mucoid Pseudomonas aeruginosa to imipenem.levofloxacin.meropenem,cefepime,cefoperazone/sulbactam,amikacin,gentamicin,piperacillin.tazobactam,ceftazidime and cefotaxime were 5.3%, 16.1%, 5.6%, 20.6%, 1.1%, 10.5%,26.9%,5.3%.31.5% and 60.2%,respectively.The drug resistant rates of mucoid Pseudomonas aeruginosa were lower than those of non-mucoid except to ceftazidime.Conclusion Compared with non-mucoid Pseudomonas aeruginosa,antibiotic resistance of mucoid Pseudomonas aeruginosa is weaker in vitro.

Objective To investigate whether oral administration of probiotics can reduce complications induced by interventional therapy in patients with primary hepatic carcinoma and liver cirrosis.Methods Two hundred and sixty four patients with primary hepatic carcinoma and liver cirrhosis who underwent transarterial chemoembolization(TACE) were randomly divided into two groups.and patients in experimental group were given probiotics but not in control group.Shoa-term clinical manifestations.liver functions,blood routine and pain scores were compared between two groups.Results On the day 3 after therapy,the incidence of abdominal distension and constipation in experimental group were less than that in control group(x2=18.22 and 55.22,P=0.000);On the day 7 after therapy,the incidence of abdominal distension,constipation and infection in experimental group were less than that in control group(x2=5.35,13.5 and 19.14,P=0.021,0.000 and 0.000).There were no significant difference in other clinical manifestations,liver function,blood routine and pain scores between the two groups. Conclusion Oral administration of probiotics can reduce the incidence of some short-term complication induced by interventional therapy in patients with hepatic carcinoma and liver cirrhosis.

Objective To investigate Smac/DIABLO and cytochrome c(cyt-c)mRNA levels in liver tissue of rats with acute hepatic failure treated by microencapsulated hepatocyte.Methods Acute hepatic failure were induced by intraperitoneal injection of D-galactosamine in rats.and the rats were treated with microencapsulated hepatocytes,free hepatocytes and physical saline(contr01),respectively.Smac/DIABLO and cyt-c mRNA in liver tissue was detected by RT-PCR and the mRNA expression levels among three groups were compared.Results Smac/DIABLO and cyt-c mRNA levels in liver tissues of rats with acute hepatic failure were higher than those of normal rata(F=4.345,14.821,47.565,42.178 and 62.961,P<0.05).The peak values of Smac/DIABLO and cyt-C mRNA expressions in free hepatocytes and control groups were at 48 h.while that in microencapsulated hepatoeytes group was at 24 h.Conclusion Smac/DIABLO and cyt-c mRNA expression is an indicator of apoptosis of hepatocytes.

Objective To investigate the genetic features related to drug-resistance of a strain of pan-drug resistant Acinetobacter baumannii. Methods The susceptibilities to 15 antibacterial agents were detected in Acinetobacter baumannii by K-B paper diffusing method. A strain of pan-drug resistant Acinetobacter baumannii was selected. The drug-resistant genes of the selected strain were amplified by polymerase chain reaction (PCR) , including 15 extended-spectrum β-lactamase genes, 3 metal β-lactamase genes, 2 AmpC enzyme genes, 7 aminoglycoside modifying enzyme genes and the genetic markers of integron I and transposons. Results The strain was resistant to 14 antibacterial agents except cefoperazone sodium/ sulbactam. β-lactamases genes ( TEM, OXA-23 and ADC) , aminoglycosides modification enzyme genes [aac(3)- I , aac(6')- I b and ant(3")- I ] , integron I qacEAi-sull and transposon tnpU were positive in PCR amplification. A new subtype of AmpC (chromosome) desoxyribonucleic acid ( DNA) was detected. Conclusion Acinetobacter baumannii is of high resistance to most antibacterial agents, and the mechanisms of the resistance are complex.

Objective To investigate the distribution of 22 beta-lactamase genes and enzyme activities in multi-drug resistant Acinetobacter baumannii (MDRAB). Methods Sixty-two MDRAB strains were isolated from the First Affiliated Hospital, College of Medicine, Zhejiang University. Twenty-two beta-lactamase genes were analyzed by PCR and verified by DNA sequencing. Enzyme activities of extended spectrum beta-lactamases (ESBLs) , cephalosporinase ( AmpC) and metallo-beta-lactamases ( MBL) were detected by the modified three-dimensional method using enzyme extraction. Results In 62 MDRABs, 51 (82.3% ) , 50 (80.6% ) and 36 (58.1% ) isolates were found to carry blaTEM, blaOXA-23 cluster, and blaADC, respectively. The rest 19 genes were not detected in this study. DNA sequencing and genomic comparison showed that 5 isolates carrying blaTEM had the same genotype as blaTEM-l , 6 isolates carrying blaOXA-23 cluster had the same genotype as blaOXA-23 carbapenemases (accession; CAB69042. 1) , and 2 isolates carrying blaADC had the same genotype as blaADC-like (accession: EU081908). Thirty-two isolates (51.6% ) produced ESBLs and AmpC, 19 isolates (30.6% ) produced ESBLs only, and 1 isolate (1.6%) produced AmpC only; and no isolate produced MBL. Conclusion MDRAB carrying blaOXA-23 carbapenemase and blaADC AmpC in this study are of high drug resistance.

Objective To investigate the clinical effect of IFNα combined with mannan peptide in treatment of patients with HBeAg-positive chronic hepatitis B ( CHB ). Methods Eighty HBeAg-positive CHB patients with HBV DNA quantity ranging from 10 to 10 eopies/mL were enrolled and randomized into the treatment group and the control group ( n = 40 for each ). Patients in treatment group were given daily subcutaneous injection of IFNα-2b 5,000,000 U for 52 weeks, and received mannan peptide 10 mg per intravenous injection or 2. 5 mg per intramuscular injection for a total of 2 to 3 treatment courses (12 weeks for each). The control group received only IFNα-2b treatment. Liver function, serum markers of hepatitis B, HBV DNA quantity and blood tests were performed before the treatment and at 2, 4, 8, 16, 26 and 52-week during the treatment; and the adverse effects were recorded. Results The rates for ALT normalization, negative HBsAg, negative HBeAg, HBeAg seroconversion and negative HBV DNA were 91. 8% , 17. 5% , 52. 5% , 27. 5 % and 47. 5% at 52nd week in the treatment group, while those in the control group were 80. 0% , 12. 5% , 30. 0% , 10. 0 % and 25. 0% , respectively. There were significant differences in HBeAg-negative, HBeAg-seroeonversion and HBV DNA-negative rates between two groups (χ2 = 4. 178, 4.021 and 4.381, P < 0. 05 ) , and these indexes in the treatment group were increased to 57. 5% , 30. 0% and 50. 0 respectively at 52nd week after drug withdraw. White blood cells began to be elevated at 4th week and were restored to the normal levels at 8th week in the treatment group, while the count in the control was lower than the normal value even at 52nd week of the treatment with the average of (3.45±1. 18)×109/L. Conclusion Alpha-interferon combined with mannan peptide therapy is effective for patients with HBeAg-positive CHB, which may restore the declined peripheral WBC counts induced by interferon and improve the compliance.

Objective To investigate the cultivation, biochemical features and drug susceptibilities of Vibrio vulnificus. Methods Three strains of Vibrio vulnificus were isolated from fatal patients in the Second Hospital of Jiaxing, Zhejiang province. Cultivation, identification and antibacterial susceptibility test were performed. Results Vibrio vulnificus grew on blood agar as dull-gray, opaque colonies with β-hemolysis. The organism presented positive in lactose, cellobiose fermentation and O/129 (10 μg) tests, but lack of inositol and rhamnus. The antibacterial susceptibility tests showed that Vibrio vulnificus strains were sensitive to ciprofloxacin, levofloxacin, imipenem, cefepime, ceftazidime, ceftriaxone, compound sulfamethoxazole and nitrofurantoin, however, resistant to gentamicin, tobramycin, aztreonam and cefazolin. Conclusions Vibrio vulnificus can be isolated from blood, bubbles fluid, and stool. Rapid identification, early diagnosis, and prompt empirical antibacterial therapy are important for reducing the mortality.

Objective To investigate the contents of IL-1β and IL-1ra in cerebrospinal fluid of children with Japanese encephalitis, and their clinical significance. Methods Enzyme linked immunosorbent assay (ELISA) was employed to detect IL-lβ and IL-lra contents in 50 children with Japanese encephalitis and 20 children without nervous system disease (controls). Results IL-1β contents in climax stage, convalescent stage and the controls were (49. 43±14. 59) , (24. 73±14. 50) and (8. 98± 1.26)μg/L (F = 79.88, P<0.01); IL-lra contents in climax stage, convalescent stage and the controls were (177. 39±60. 19), (78. 24±44. 63) and (21. 09±3. 10) μg/L (F = 91. 53, P <0. 01). There were significant differences on IL-lβ and IL-lra contents among children with mild, moderate and severe encephalitis (climax: F = 82.36 and 66.50, P<0.01; convalescence; F = 55. 17 and 79.50, P<0.01). IL-1β content was positively correlated with IL-lra in both climax and convalescent stages (climax; r = 0. 815, P < 0.01; convalescent; r= 0.728, P < 0.01). Conclusions IL-lβ and IL-lra contents in cerebrospinal fluid are significantly increased in children with Japanese encephalitis in climax stage, which are closely correlated with the disease severity. The two indicators may participate in the pathological process of brain damages with Japaness encephalitis.