Humans ; Interleukin-33 ; Interleukins ; Liver Cirrhosis

Objective To investigate the clinical and epidemiologieal feat ures of epidemic cerebrospinal meningitis in recent 3 years,aiming to develop the strategies for controlling this disease.Methods A retrospective analysis was conducted in 65 hospitalized patients with epidemic cerehrospinal meningitis from 2003 to 2006.Chi-square test was used for statistical analysis.Results The majority of these 65 patients were juvenile and adult,accounting for 44.6%(29 cases)and 35.4%(23 cases),respectively,while the infant patients account for the smallest percentage,only 6.2%(4 cases).Most cases occurred in spring(from February to April).The clinical features of most patients belonged to common type(72.3%),followed by fulminant type(27.7%).Five cases(7.7%)died,all of whom were fulminant cases.Totally 33 strains of Neisseria meningitidis were isolated,with positive culture rate of 50.8%(33/65 cases).The positive rate of blood culture was 44.6%(29/65 cases)and that of cerebrospinal fluid cuhure was 31.4%(16/51 cases).Isolates of Neisseria meningitidis were still highly sensitive to penicillin,ceftriaxone and cefotaxime.However,the resistance rates of strains to compound sulfamethoxazole,gentamycin and ciprofloxacin were all above 80%.Resuhs of serological typing revealed thai 82.8%(53/64 cases)cases belonged to group C.There were more severe cases and higher death rate in patients infected by group C meningococcus.Conclusions Serogroup C of the meningococcus have become the preponderant strains in Anhui Province.Most of the patients infected hy group C are J uveniles and adults.Penicillin and the third generation cephalosporins are still highly aclive againsl Neisseria meningitidis.

OBJECTIVE To investigate the distribution and the resistance rate of Shigella in Anhui Province to guide the choice of antibacterials. METHODS Ninety one strains of Shigella were cultured in Sep 2005.The groups were identified by biochemical and serologic tests.Susceptibility of 91 strains of Shigella in Anhui to various antibiotics was tested using standardized custom dilution MIC panels according to CLSI(2005) guidelines. RESULTS There were 57 strains of Shigella flexneri,31 strains of S.sonnei and 2 strains ofS.boydii among 91 strains of Shigella.The resistance rates of Shigella to cefoperazone/sulbactam and piperacillin/tazobactam were remarkably lower than to other third generation cephalosporins.The susceptible rates to carbopenems were 100%.The resistance rates to ciprofloxacin lactate and pazufloxacin mesilate were 27.47% and 32.97%,respectively. CONCLUSIONS There is a certain resistance rate of the Shigella to fluoroquinolones and the third generation cephalosporins.More attention should be paid to the surveillance and control of such resistance.

Objective To investigate the inhibitory effect of aqueous extract of Taxus chinensis.vat (AETC) combining Cisplatin (DDP) on vitro cultured human lung carcinoma A549 cells,and the effects on resistance genes.Methods The A549 cells were divided into different concentrations of DDP groups,different concentrations of AETC groups,and blank group,and drug effect of 48 h with the method of cell counting kit-8 (CCK-8) and the effect on cell survival were detected.Based on the above results,then A549 cells were divided into DDP combining different concentrations of AETC groups,DDP group,blank control group,and drug effect of 48 h with the method of CCK-8 and the effect on cells survival were detected.The gene expressions of adenosine triphosphate (ATP)-binding cassette subfamily B member 1 trans-porter (ABCB1),ABCG2,and ABCC1 were examined by polymerase chain reaction (RT-PCR).Results Cisplatin 12 μg/ml (DDP),DDP + ATEC 400 μg/ml,DDP + ATEC 800 μg/ml,DDP + ATEC 1 200 μg/ml,DDP + ATEC 1 600 μg/ml,A549 cell inhibition rate of each group was 44.36％,69.61％,74.73％,80.10％,and 74.73％,respectively;Different concentrations of AETC combining DDP could decrease the resistance related gene ABCC1,ABCB1 expressions,and correlated to the dose.AETC combining DDP showed no effects on ABCG2 gene expression.Conclusions AETC combining DDP could inhibit the growth of A549 cells,and decrease the resistance-related gene ABCC1,ABCB1 expressions.

Aim To obtain the encoding gene sequences of TEM-type ?-lactamases produced by 4-strain Klebsiella pneumoniae in Zhejian g Province, identify their genotypes and study some properties of these TEM-typ e ?-lactamases.Methods The encoding genes of TEM-type ?-la ctamases produced by 4 isolates were amplified by PCR. The purified PCR products were ligated with pGEM-T easy vectors, expressed in E. coli DH 5?, and sequenced by Sanger's dideoxy chain termin ation composition method. Compared with anino acid sequences in the GenBank,TEM -types of the ?-lactamases was determined. The genes of TEM ?-lactamases were ligated with pET-28 c vector to express recombinant proteins in E. coli DH 5?. Plasmids were extracted from the p ronucleus expression strains and PCR was performed to determine whether the pron ucleus expression was successful or not. Their phenotypes were determined by ESB Ls phenotype affirmative test. The isoelectric points (pIs) of the recombinant p roteins were determined by isoelectric focus. Conjugation test was performed to determine whether their genes existed in plasmid or chromosome. Results The encoding genes of ?-lactamases were determined as TEM by PCR. It s PCR product had 1 009 nucleotides. The pI of the novel TEM ?-lactamase was 5.4. The enzyme was determined as non-ESBLs by ESBLs phenotype affirmative tes t.Transconjugants were successfully selected from the paternal producers in conj ugation tests. The TEM-type ?-lactamase produced by 4 strains was determined as TEM-105(AF516720) by GenBank. Conclusion The ?-lactamase produced by 4-strain K. pneumoniae from 4 patients in Zhejiang Province was TEM-105. It was the first report of TEM-105 type ?-lactamase produced by 4-st ain K. pneumoniae from China in the world.

Objective To establish a real-time quantitative polymerase chain reaction (RQ-PCR) assay for fast detection of Aspergillus fumigatus genome in human whole blood samples and explore its clinical application.Methods The primers and the TaqMan-probe were designed on the basis of the multi-copy ITS1-5. 8S region of the rDNA of Aspergillus fumigatus. The Aspergillus fumigatus genomic DNA were extracted with QIAamp(R) DNA Blood Mini Kit.A 20 μl RQ-PCR amplification system was established, and the simulated blood samples containing various given load of Aspergillus fumigatus genome and the 66 whole blood samples of the surgical febrile patients were examined. Results The detection limit of the RQ-PCR instrument is 10-1 genomes/μl DNA sample,namely 78 CFU/ml whole blood. The specificity and the sensitivity were 94. 25% and 99. 04% respectively; and the positive predictive value and negative predictive value were 97. 63% and 97. 62% respectively. The average relative error of the quantitative results was (3. 67 ±13. 19)%, and the intra- and the inter-assay average coefficients of variation were (12.38 ± 1. 53)% and (16. 27 ±2. 72)% , respectively. The average recovery rate of Aspergillus fumigatus genomic DNA in human whole blood samples was (107. 81 ±25. 92)% , and the average coefficient of variation of the average recovery rate was (26. 24 ± 5.62) % . No Aspergillus fumigatus genomic DNA was detected among the 66 blood samples of the surgical febrile patients. Conclusions The RQ-PCR assay for fast quantitative detection of Aspergillus fumigatus genome in human whole blood samples is of high sensitivity, specificity,accuracy and precision. The Aspergillus fumigatus genome was not detected in this group of surgical febrile patients.

Objective To detect the integration of hepatitis B virus (HBV) DNA in HBVrelated human hepatocellular carcinomas (HCC). Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction (PCR) with specially designed U-base primers. According to the known genes and human Alu repeat sequences (Alu repeat) , primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies.Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information (NCBI) basic local alignment search tool (BLAST) and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn't show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

Objective To investigate the expressions of hepatorna stem cell surface marker CD133 CD90 in tissues of hepatocellular carcinoma (HCC) and evaluate their related clinical significances. Method The expressions of CD133 CD90 were detected by immunohistochemical method in HCC tissues of 93 patients, and normal liver tissues of 10 cases. Results Among 93 cases with HCC, the positive expression of CD133 were found in 71 cases (76.3%), and CD90 positive expression in 64 cases (68.8%), and the percentage of positive cells were (6.4±3.3)% and (4.3±3.9)% respectively. No positive expression of CD133 and CD90 was found in normal liver tissues (P<0.01). CD133, CD90 expressions in the HCC tissues of TNM stage Ⅲ-Ⅳ [(8.1±3.7)%,(5.7±4.2)%] were higher than those of TNM stage Ⅰ - Ⅱ [(4.1±2.3)%,(2.3±1.9)%] (P<0.01). Spearman correlation analysis indicated that the expressions of CD133 and CD90 were up-regulated as the pathohistology grades increased (P<0.05). Positive correlation was observed between CD133 and CD90 expression (r=0.402, P<0.01). Conclusions CD133, CD90 positive cells exist in HCC tissues, their expressions positively relate to the TNM stage and the pathohistology grades for HCC patients.

Objective To study the methylation status of secreted frizzled-related protein (SFRP) 1 and SFRP2 genes in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and the relationship between the methylation status of the two genes and the development of HCC.Methods Using methylation-specific polymerase chain reaction (MSP) to detect methylation status of SFRP1 and SFRP2 genes of 45 specimens of HCC tissue and adjacent non-tumorous liver tissue from HCC patients during operations,and 6 normal liver tissues from patients with cholecystolithiasis or hepatic hemangiomas. The data were analyzed by chi-square test and Fisher exact test. Results SFRP1 gene methylation was detected in 28 HCC tissues and 16 adjacent non-tumorous liver tissues,accounted for 62.2% and 35.6%,respectively;and SFRP2 gene methylation was detected in 23 HCC tissues and 13 adjacent non-tumorous liver tissues,accounted for 51.1% and 28.9%,respectively;while no methylation was detected in 6 samples of normal liver tissues. There was no significant difference between the methylation of SFRP1 and SFRP2 genes in HCC tissues and gender,age,HBV serum markers,types of adjacent non-tumorous liver tissues,metastasis and pathological stage (P＞0.05).The abnormal methylation status between SFRP1 and SFRP2 genes was linear correlated in HCC tissues (r=0.381,P=0.01).Conclusion Hypermethylation of SFRP1 and SFRP2 genes frequently occurs in HBV-related HCC,which may be an important molecular biomarker for prediction of hepatocarcinogenesis in the future.

Objective To investigate platelet function with special regard to the role of platelet membrane glycoproteins (GPIIb/IIIa?GPIb) and endothelial cell disturbance in the older non valve cardiac patients with atrial fibrillation and their clinical implications. Methods 22 older patients with non valve cardiac atrial fibrillation were studied. There were two age and sex matched control groups, one with 18 patients in sinus rhythm with cardiovascular disease, called the normal sinus rhythm group; the other with 16 health subjects named the health control group. The expression of activated GPIIb/IIIa and GPIb on platelet was analyzed with flow cytometry. Mean platelet volume (MPV) was measured by automatic hematologic analyzer, the plasma vWF was assayed using ELISA. Results The non valve atrial fibrillation group had markedly activated platelet, indicated by increased expression of activated GPIIb/IIIa ( P