Objective To explore the clinical effects and mechanisms of Hyper-IL-6 mediated by recombinant adenovirus on rat with acute liver failure(ALF).Methods Eighty ALF rats were randomly divided into 4 groups, normal saline was injected for non-infection group;Ad0,AdIL-6 and AdHIL-6 was respectively injected in group Ad0,group AdIL-6,group AdHIL-6.Subsequently the index of hepatic function, liver pathology, hepatocelluar apoptosis in situ and the expression of PCNA and caspase-3 in liver tissue were examined. The survival rate of rats for 14 days was recorded.Results The level of serum ALT and TB,and apoptotic index in rats of group AdHIL-6 was markedly decreased but expression of PCNA increased (P

Objective To explore the expression and significance of vascular endothelial growth factor (VEGF) and transforming growth factor β1 (TGF-β1) in hepatic fibrosis.Methods Fifty-six cases of patients with hepatic fibrosis were selected as observation group and 50 healthy persons as control group.Immunohistochemistry were performed to detect VEGF and TGF-β1 in two groups.Results Serum VEGF and TGF-β1 in observation group were significantly higher than those in control group(VEGF:(110.87 ±32.64) μg/L vs (15.98 ±6.75) μg/L,t =20.166,P ＜0.001;TGF-β1:(15.08 ±4.27) ng/L vs (7.17 ±2.86) ng/L,t =11.066,P ＜ 0.001) ;There were significant differences on VEGF and TGF-β1 level among S1,S2,S3 and S4 subgroups(VEGF:(84.25 ±16.86) μg/L vs (101.87 ±36.70) μg/L vs (118.04 ±40.75)μg/L vs (134.65 ± 45.73) μg/L,F =15.689,P =0.015 ; TGF-β1:(10.87 ± 2.64) ng/L vs (13.06 ± 2.74)ng/L vs (17.87 ± 3.28) ng/L vs (22.76 ± 4.75) ng/L,F =12.438,P =0.026).VEGF had positive correlation with TGF-β1 (r =0.532,P =0.013).Conclusion VEGF and TGF-β1 level have close relationship with the occurrence and development of hepatic fibrosis.Combined detection of VEGF and TGF-β1 can be serum index for diagnosis and evaluation disease condition.

In order to assess the significance of human malignancy-associated nucleolar antigen P24 in the diagnosis of tumors,552 specimens of human tissues were stained and observed with immuno - fluorescent and ABC methods using anti-P24 monoclonal antibody MA2.It was found that P24 was positive in the nucleolus of 97% cases of malignant tumors and in 50% of atypical hyperplastic tissues but negative in 98.6% cases of benign tumors and in all the normal or nontumorous tissues.On the basis of these findings,it is believed that P24 is highly specific to malignant tumors and will be a useful marker for the differential diagnosis between malignant and benign tumors.

To investigate the distribution of the genes of two major metallo-beta-lactamases (MBL; i.e., IMP and VIM) and class 1 integrons (intI) in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for bla ( IMP-1 ), bla ( VIM ) and bla ( VIM-2 ) genes. The MBL-positive isolates were further assessed for class 1 integrons by PCR using specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the bla ( VIM ) gene was found in 81.5% (53/65) of all isolates, bla ( VIM-2 ) gene was found in only 1 isolate and the intI gene was observed in 45.3% (24/53) of bla ( VIM )-positive isolates. One isolate carried simultaneously both bla ( IMP-1 ) and intI genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM beta-lactamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P. aeruginosa.
Anti-Bacterial Agents/pharmacology ; China ; DNA Primers/chemistry ; Drug Resistance, Bacterial ; Gene Expression Regulation, Bacterial ; Imipenem/*pharmacology ; Integrons ; Microbial Sensitivity Tests ; Models, Genetic ; Pseudomonas Infections/genetics ; Pseudomonas Infections/*microbiology ; Pseudomonas aeruginosa/*metabolism ; Sequence Analysis, DNA ; beta-Lactamases/*metabolism

Objective To assess the clinic application of a new-style automated immunoassay system HISCL ( high sensitivity chemiluminescent enzyme immunoassay ) with high sensitive chemiluminescence substrate CDP-Star?, bind-free separate technology and filter conversion technology.Methods The performance verification test evaluated HISCL′s specification including reproducibility, functional sensitivity, linearity, accuracy, and sensitivity for HBsAg seroconversion.To evaluate the specificity , 1 007 HBsAg-negative specimens , 82 potentially interfering specimens were from conventional specimens in West China Hospital , Sichuan University between October and December of 2014.At the same time, with these results to determine the rate of the specimens that their results are in negative grey zone.259 HBsAg-positive specimens and 27 weakly-positive specimens were tested for the comparison between the HISCL and ECLIA ( electrochemiluminescence immunoassay ) HBsAg quantification.A linear regression model , Pearson′s correlation and Bland-Altman analysis were used as statistical methods.Results The between day and within-lot variable coefficient of two level samples are 1.55%, 2.02%, 0.34%, 1.34%separately.The functional sensitivity of the HISCL HBsAg assay reaches 0.007 IU /ml.There is a good linearity (y=3.262x+0.082,r=0.994; y=2 303.608x-33.006,r=0.999) within range of 0-2 300 IU/ml.HISCL obtain the accurate results for the CAP control material , the agreement rate is 100%.The sensitivity for the detection of HBsAg seroconversion is high.The specificity is 99.91%.The negative grey zone rate is 1.66%.The results of these two methods have good correlation and uniformity , r=0.995, LOA:-0.3 -0.19 log10 IU/ml.Conclusions HISCL HBsAg detection system shows good detection performance.Be of the function of both qualitative and quantitative detection.And the negative “grey zone”specimen rate is low.It is wholly capable of wide linearity and quick assays for quantifying serum HBsAg levels.HISCL could contribute to improve HBsAg quantitative detection process for clinical laboratory.

Objective:To clone the cDNA in full-length of human mammaglobin,do prokaryotic expression and purify hMAM protein,as a basis for early diagnosis of breast cancer.Methods :hMAM cDNA was amplified through RT-PCR from breast cancer tissue and breast cancer cell line MD-MB453,and the recombination pQE40-hMAM vector was constructed and expressed in E.Coli.M15 after induction by IPTG.The fusion protein was purified with Ni-NTA-His affinity chromatography. Results: Two subtypes of hMAM cDNA and the hMAM(Isoform) cDNA were found,in which consisted of 270 bp,different from the wildtype hMAM cDNA of 279 bp on nine continuous base pair missing.The fusion protein formed inclusion body in prokaryotic expression system and the renatured protein was purified which purity was about 97%.Conclusion: The recombinant hMAM was successfully purified.

Objective To analyze the clinical performance of chemiluminescence immunoassay (CLIA)in determination of trepo‐nema pallidum antibody(TP antibody) .Methods The results detected by enzyme‐linked immunosorbent assay( ELISA)were regar‐ded as relative standards ,and results detected by treponema pallidum particle assay (TPPA) were regarded as recognition criteria . 2 223 serum samples of outpatients and inpatients were collected ,and TP antibodies were detected by CLIA and ELISA method re‐spectively ,and followed by confirmation of TPPA test .Results Among 2 223 serum samples ,53 samples were TP antibody positive detected by ELISA and 60 samples were TP antibody positive detected by CLIA ,and the positive incidence of TP antibody detected by the ELISA and CLIA method was 2 .34% and 2 .65% respectively .The positive predictive value ,sensitivity and specificity of the CLIA method was 98 .33% ,100 .00% and 99 .95% ,repectively .Conclusion The CLIA method could be considered adequate for screening of TP antibody in a large volume of samples ,with characteristics of automatic ,quantitative and short turn around time .

OBJECTlVE To study the Iong term toxicity of vinoreIbine tartrate(NVB)on rat immune and hematopoietic systems pathoIogicaIIy. METHODS SD Rats were randomIy divided into 4 groups:normaI controI group and NVB 5.0,10.0,and 20.0 mg·m-2 groups,each group containing 6 maIe and femaIe rats. The rats in NBV groups were administered different concentrations of NVB by intravenous drip on the 1st and 8th days,21 da cycIe,for 4 cycIes. On the 14th day after the Iast administration, white bIood ceIIs(WBC),neutrophiI(Neut),Iymphocytes(Lym),red bIood ceIIs(RBC)and reticuIo-cyte‰(RET‰)were detected by ADVIA2120 hematoIogy anaIyzer. Thymus,sternum marrow,spIeen and mesenteric Iymph nodes were observed by histopathoIogicaI examination. The thymus and spIeen were preciseIy weighed to obtain the reIative organ coefficients. Bone marrow smears were made for counting and cIassification. RESULTS Compared with normaI controI group,WBC,Neut,Lym,RBC and RET% of peripheraI bIood of NVB 5,10 and 20 mg·m-2 groups were decreased(P﹤0.05,P﹤0.01). The Neut vaIue of maIe rats was(2.35±0.56)×109·L-1 in normaI controI group,but was reduced to (1.66±0.44),(0.67±0.22)and(0.20±0.02)×109·L-1(P﹤0.05,P﹤0.01)in NVB 5,10 and 20 mg·m-2 groups. The Neut vaIue of femaIe rats was(1.26± 0.27)× 109 L-1 in normaI controI group,but was reduced to(1.14±0.56),(0.47±0.13)and(0.21±0.08)×109 L-1(P﹤0.05,P﹤0.01)in NVB 5,10 and 20 mg·m-2 groups. The resuIts of counting and cIassification of bone marrow smears showed that the myeIoid ceII ratio decreased(P﹤0.05,P﹤0.01). The myeIoid ceII ratio of maIe rats was(42.7±6.1)% in normaI controI group,but was reduced to(28.8±5.3)%,(22.0±3.2)% and(18.9±3.9)% in NVB 5,10 and 20 mg·m-2 groups. The myeIoid ceII ratio of femaIe rats in normaI controI group was(35.4±3.0)%, but was reduced to(31.2±4.7)%,(22.9±6.7)% and(20.8±4.2)% in NVB 5,10 and 20 mg·m-2 groups. The thymus coefficient was reduced(P﹤0.05,P﹤0.01). The thymus coefficient of maIe rats in normaI controI group was 0.36±0.04,but was reduced to 0.31±0.06,0.18±0.03 and 0.08±0.01 in NVB 5,10 and 20 mg·m-2 groups. The thymus coefficient of femaIe rats in normaI controI group was 0.29±0.06,but was reduced to 0.25±0.06,0.19±0.06 and 0.07±0.01 in NVB 5,10 and 20 mg·m-2 groups. Histopatho-IogicaI examination showed that thymus was atrophiedand bone marrow was suppressed. SpIeen com-pensatory extrameduIIary hematopoietic ceIIs were increased in NVB 5.0,10.0 and 20.0 mg·m-2 groups (maIe and femaIe)to different degrees,but the mesenteric Iymph nodes of NVB groups showed no sig-nificant pathoIogicaI changes. CONCLUSlON NVB has immune and hematopoietic toxicity on SD rats, as is showed by thymic atrophy and bone marrow suppression.

To investigate the distribution of the genes of two major metallo-β-lactamases (MBL; i.e., IMP and VIM) and class 1 integrons (intI) in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for blaIMP-1, blaVIM and blaVIM-2 genes. The MBL-positive isolates were further assessed for class 1 integrons by PCR using specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the blaVIM gene was found in 81.5% (53/65) of all isolates, bla gene was found in only 1 isolate and the intI gene was observed in 45.3% (24/53) of blaVIM-positive isolates. One isolate carried simultaneously both blaIMP-1 and intI genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM β-lactamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P. aeruginosa.

OBJECTIVETo analyze the distribution of Enterobacteriaceae isolated from West China Hospital, investigate the antibiotic resistance profile of Enterobacteriaceae with decreased susceptibility to carbapenems and explore the molecular mechanism.

METHODSForty-five Enterobacteriaceae strains resistant or with reduced susceptibility to carbapenems were isolated from patients in West China Hospital. The antimicrobial susceptibility and carbapenemase-producing phenotypes of the bacteria were examined and specific PCR were performed to determine the molecular mechanism.

RESULTSOf the 45 isolates, 17, 21 and 36 were resistant or intermediate strains to imipenem, meropenem and ertapenem, respectively. The majority of these isolates showed resistance to cephalosporins. The modified Hodge test resulted in the highest positivity rate (77.8%), followed by EDTA disc test (57.8%) and PBA disc test (22.2%). BlaTEM, blaSHV and blaCTX-M were detected in 60.0%, 53.3% and 15.6% of these strains with reduced susceptibility. The rate of strains carrying 2 or more genes was 44.4%, and the detection rate of blaIMP was 48.9%. BlaKPC was identified in 4 (8.9%) high-level resistant strains and confirmed to locate on the plasmid.

CONCLUSIONProduction of carbapenemase contributes to reduced susceptibility of carbapenems in Enterobacteriaceae. The presence of blaKPC, MBL and ESBL, and their possible combinations can be the main factor contributing to carbapenem resistance or reduced susceptibility in Enterobacteriaceae. The KPC-2 carbapenemase gene located on the plasmids we found in this study can cause potential horizontal transmission across strains.

Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Carbapenems ; pharmacology ; Cephalosporins ; pharmacology ; Enterobacteriaceae ; drug effects ; enzymology ; genetics ; Gene Amplification ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Polymerase Chain Reaction ; Thienamycins ; pharmacology ; beta-Lactam Resistance ; beta-Lactamases ; genetics ; metabolism ; beta-Lactams ; pharmacology