ObjectiveToinvestigatetheplasmidgenechangesinquinolone‐sensitivePseudomonasaeruginosa.Methods 31iso‐lates from January 2011 to December 2013 from various qualified clinical samples in the hospital were collected .In the 31 isolateds , 16 isolates proved sensitive to quinolones by using K‐B method were used as research objects in the study .The isolates growing ou‐side the sensitive ring of ciprofloxacin paper were selected to continuously transferred into other culture dishes until the resistance to quinolones were acquired .Plasmid transformation and extraction were performed on those isolates to confirm the existence of drug‐resistanceplasmidsacquired,andthroughPCRandgenesequenceanalysistodeterminethetypeofplasmids.Results 2iso‐lates of quinolone‐sensitive Pseudomonas aeruginosa acquired drug‐resistance plasmids qnrS and were resistant quinolones induced by continuous transferring for 9 times .Conclusion If antibiotics of inhibitory concentration were often used for the treatment of Pseudomonas aeruginosa infection ,drug‐resistance plasmids were acquired easily .

Adolescent ; Biopsy ; Cystic Adenomatoid Malformation of Lung, Congenital ; diagnosis ; diagnostic imaging ; therapy ; Diagnosis, Differential ; Diagnostic Errors ; adverse effects ; prevention & control ; Humans ; Infant ; Lung Diseases ; diagnosis ; diagnostic imaging ; therapy ; Male ; Middle Lobe Syndrome ; diagnosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo explore the relationship between body mass index, waist circumference and blood pressure among residents in Chongqing area.

METHODSA total of 5246 residents aged 15 and over in Chongqing area were enrolled in this study by use of stratified sampling and cluster sampling methods. Data on blood pressure (SBP, DBP), pulse, height, body weight, waist and hip circumferences as well as questionnaire survey were analyzed.

RESULTSThe level of SBP and DBP and hypertension prevalence rate were significantly positively correlated with BMI (all P < 0.01). SBP, DBP levels and hypertension prevalence rate were significantly higher in people with abdomen obesity than people with normal waist circumference (all P < 0.01). BMI, waist circumference in hypertensive residents were significantly higher than non-hypertensive residents (all P < 0.01).

CONCLUSIONBlood pressure level and hypertension prevalence rate were closely related with BMI and waist circumference among residents in Chongqing area.

Adolescent ; Adult ; Aged ; Blood Pressure ; Body Mass Index ; China ; epidemiology ; Female ; Heart Rate ; Humans ; Hypertension ; epidemiology ; physiopathology ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Rural Population ; Urban Population ; Waist Circumference ; Waist-Hip Ratio ; Young Adult

Human tumstatin(hTumstatin)cDNA was amplified from recombinant plasmid pET-3c-tum, cloned in frame with the signal sequence in yeast vector pPICZalphaA and transformed into Pichia pastoris GS115 by electroporation. The expression of hTumstatin in GS115(pPICZalpha-tum)was then induced by methanol and secreted into the culture medium, with a yield of 25mg/L as shown by SDS-PAGE and Western blotting. The expressed hTumstatin was purified to more than 85% purity using a simple one-step SP-Sepharose cation exchange chromatography. The MTT and chick chorioallantoic membrane assay showed that the yeast produced hTumstatin could inhibit the proliferation of human umbilical vein endothelial cells and the neovascularization induced by bFGF. Hoechst 33258 fluorescent staining also demonstrated the apoptotic change in endothelial cellular nuclear morphology.
Angiogenesis Inhibitors ; metabolism ; Autoantigens ; genetics ; metabolism ; Cell Proliferation ; Cells, Cultured ; Collagen Type IV ; genetics ; metabolism ; DNA, Complementary ; genetics ; Electroporation ; Endothelial Cells ; cytology ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Umbilical Cord ; cytology

OBJECTIVETo investigate the related to relapse of chronic hepatitis B (CHB) after recombinant interferon-alpha (rIFN-alpha) treatment.

METHODSThis investigation involved 523 pathologically confirmed CHB patients including 403 HBeAg-positive and 120 HBeAg-negative patients, who were treated with 5 MU rIFN-alpha subcutaneously thrice a week for 6-25 months. For each patient, serum alanine aminotransferase (ALT) was measured biochemically, serum HBV DNA level detected with quantitative fluorescent PCR, and HBeAg level with enzyme immuoassay every 1-3 months during therapy and every 3-6 months during the follow-up period.

RESULTSEarly response to rIFN-alpha treatment was observed in 302 (57.7%) patients at the end of treatment, among whom 39.4% (119/302) suffered relapse during the follow-up for 39.2-/+21.5 months. Age, HBeAg status before treatment, and follow-up duration were the predictive factors for post-treatment relapse. The mean age of patients with CHB relapse was significantly higher than that of the sustained responders (P<0.001), and the relapse rates in HBeAg-negative group (55.8%, 43/77) were significantly higher than that in HBeAg-positive group (33.8%, 76/225) at the end of follow up (P<0.001). The relapse rate and accumulative relapse rates at each year during the follow-up (for 5 years as the longest) differed significantly (P<0.001, P=0.000), but the accumulative relapse rates differed little between the years after the initial 2 of the follow-up (P=0.670). The relapse was not related to the patient's gender, pretreatment serum ALT, HBV DNA, grade of liver inflammation, stage of liver fibrosis, or duration of treatment. In HBeAg-positive patients, however, the mean HBV DNA was significantly higher in relapse group than in sustained response group (P=0.017).

CONCLUSIONAge, pretreatment HBeAg status, and follow-up duration are independent predictive factors for post-treatment CHB relapse. In HBeAg positive patients, pretreatment serum HBV DNA is also one of the risk factors for relapse.

Adult ; Age Factors ; Alanine Transaminase ; blood ; DNA, Viral ; blood ; Female ; Follow-Up Studies ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Logistic Models ; Male ; Recurrence ; Treatment Outcome

OBJECTIVETo assess the correlations between Survivin and vascular endothelial growth factor (VEGF) in hepatocellular carcinoma (HCC) and their clinical significance.

METHODSThe expressions of Survivin and VEGF in 50 HCC specimens and 20 normal hepatic tissue specimens were detected by immunohistochemistry, and the results were analyzed in relation to the patients' clinicopathologic characteristics.

RESULTSOf the 50 HCC specimens, 32 (64.0%) were positive for Survivin expression, and 34 (68.0%) were positive for VEGF expression. Survivin expression was not detected in normal hepatic tissues, and 2 (10%) of these tissues were positive for VEGF, showing significant difference in Survivin and VEGF expressions between HCC specimens and normal hepatic tissues. Survivin and VEGF expressions were not correlated to the patients' gender, age, tumor size, degree of differentiation and alpha fetoprotein level (P<0.05), but related to the clinical stage and lymph node metastasis of HCC (P<0.05). Correlation analysis indicated a close correlation between the expressions of survivin and VEGF in the HCC specimens (chi 2=6.69, P<0.05).

CONCLUSIONSurvivin and VEGF are over-expressed in HCC tissues, and may theoretically serve as the targets of molecular targeted drugs. Detection of the expressions of Survivin and VEGF in HCC tissues may provide assistance for prognostic evaluation of the patients.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Liver Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microtubule-Associated Proteins ; biosynthesis ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor A ; biosynthesis

OBJECTIVETo study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

METHODSDNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.

RESULTSOnly the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.

CONCLUSIONThe results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.

Animals ; Antigenic Variation ; COS Cells ; Cercopithecus aethiops ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Plasmids ; genetics ; Point Mutation ; Transfection

OBJECTIVETo investigate the prevalence of three lamivudine-resistant HBV mutants in lamivudine-treated Chinese patients.

METHODSUsing three pairs of of HBV polymerase gene B and C domain fragments were amplified by PCR. The PCR products were then digested by restriction enzyme Nde I and Nla III. The digested products were analyzed by electrophoresis. With this method, the prevalence of the three lamivudine-resistant mutants in lamivudine-treated Chinese patients was investigated.

RESULTSAfter Nde I digestion of p24 and p29 amplified product, HBV wild type could be easily separated from YMDD mutant. At the same time, YIDD could be separated from YVDD mutant after Nla III digestion of p24 and p29 amplified product. By this method, the authors found that these eleven patients were infected with lamivudine-resistant mutants. Six of them were infected with M5501 mutant; five were infected with M550V mutant (one of them had both M550V and L526M mutations).

CONCLUSIONThe method of the present study was demonstrated to be an easy way to detect HBV lamivudine-resistant mutants and can be applied to clinical monitoring of lamivudine resistance.

Adolescent ; Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA Mutational Analysis ; DNA Primers ; genetics ; DNA, Viral ; analysis ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction

Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; virology ; Humans ; T-Lymphocytes, Cytotoxic ; immunology

Kringle 1-3 domain is a recently found angiogenesis inhibitor with anti-angiogenesis and anti-tumor activity. The kringle 1-3 gene was amplified by PCR technique using angiostatin gene as template. After DNA sequencing, the PCR product was cloned into pPIC9K resulting in recombinant plasmid pPIC9K13 which was then transformed into Pichia pastoris GS115. The high copy integration transformants screened by PCR and G418 methods were cultivated in flasks. The K1-3 was expressed and secreted to the medium and has immunogenic activity as shown by SDS-PAGE and Western blotting. High cell density culture was carried out in 30-liter and 80-liter bioreactor, the biomass reaches 300 OD after methanol induction, and the expressed product is 200 mg/L. The fermentation supernatant was purified by Streamline SP and Phenyl Sepharose Chromatography, the product appears as a single band on SDS-PAGE, with a purity of 95%-96%. The purified product has anti-angiogenesis and anti-tumor activity.
Bioreactors ; Cloning, Molecular ; Fermentation ; Humans ; Kringles ; genetics ; Pichia ; genetics ; Plasmids ; Plasminogen ; genetics ; isolation & purification ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology