Objective To investigate the correlation between vitamin D and systemic sclerosis(SSc).Methods The serum vita-min D2,vitamin D3 and total vitamin D levels were detected in 56 outpatients and inpatients with SSc(SSc group)and 60 individuals of healthy physical examination(control group)from January 2009 to December 2013.The detection results combined with the clini-cal data were statistically analyzed.Results 3 kinds of vitamin D levels in the SSc group were lower,than control group in which the total vitamin D and vitamin D3 levels were significantly lower than those in the control group(P <0.05).Conclusion Maintai-ning the higher concentration of serum vitamin D may have the preventive effect on progressive SSc.

Objective To investigate the distribution of β-lactamase genes in a pan-drug resistant Klebsiella pneumoniae isolate JM45.Methods Klebsiella pneumoniae JM45 was isolated from the blood sample of a patient admitted in the intensive care unit,the Second Affiliated Hospital,Zhejiang University School of Medicine on April 7,2010.The susceptibilities to 26 antibiotics were tested using E-test method.Cica-β-Test was performed to detect β-lactams,and modified Hodge test was performed to detect carbapenemase.Resistant genotypes were detected using PCR,DNA sequencing and BLAST algorithm.Whole genome sequencing (complete graph) was performed by high throughput Roche 454 sequencing approach to analyze the distribution of β-lactamase genes.Results Except polymyxin B and tigecycline,JM45 was resistant to other 24 kinds of antibiotics including cephalosporins and carbapenems.Several β-lactamases were positive in Cica-β-Test,and modified Hodge test was positive.Based on PCR typing,TEM-1,SHV-11,CTX-M-24 and VEB-3 were positive,but carbapenemase genes and metallo-β-lactamase genes were negative.A complete genome (chromosome) sequence (GenBank accession number:CP006656) and 2 plasmids sequences (GenBank accession number:CP006657,CP006658) were obtained by wholegenome sequencing.CTX-M-24 (Locus tag:N559_5233),TEM-1 (Locus tag:N559_5242) and VEB-3 (Locus tag:N559_5248) were positive in plasmid 1.CTX-M-24 located in insertion sequence (IS903-CTXM-24-ISEcp1),while TEM-1 and VEB-3 located in transposons (tnpA-TEM-1-rmtB and VEB-3-tnpA).SHV-11 (Locus tag:N559_2715) was positive in genome (chromosome),and 4 putative β-lactamase genes or β-lactamase domains were obtained:(1) metallo-β-lactamase domain protein (Locus tag:N559_0119,780 bp) ; (2) putative β-lactamase (Locus tag:N559_1633,1308 bp) ; (3) β-lactamase domain protein (Locus tag:N559_2279,813 bp); (4) β-lactamase domain protein (Locus tag:N559_3769,1101 bp).No insertion sequence or transposase gene was observed near SHV-11.Conclusion The resistance to antibiotics including cephalosporins and carbapenems is correlated with TEM-1,SHV-11,CTX-M-24,VEB-3 and 4 kinds of putative β-lactamase genes or β-lactamase domains.

OBJECTIVE:To study the analgesic and anti-inflammatory effect of Shulitong granule.METHODS:The mice pain model was established by acetic acid;Dimethylbezene was used to make the external ears of the mice swollen and form cotton ball-shaped granuloma in rats so as to form models;The analgestic effect and anti-inflammatory effect of the Shulitong granule were observed respectively.RESULTS:Shulitong granule could reduce the writhing response times signifi?cantly(P

Objective To investigate ehloramphenicol,tetracycline,rifampicin and trimethoprimsulfamethoxazole resistance and the related genes in multi-drug resistant Acinetobacter baumannii(MDR-ABA).Methods Sixty-two strains of MDR-ABA were isolated from clinical samples,and their susceptibilities to 22 antimicrobial agents were detected by Kirby-Bauer disk diffusion tests.Genes related to chloramphenicol(catB and cmlA),rifampicin(arr-2/3),tetracycline(tetA and tetB)and trimethoprimsulfamethoxazole(sull,dfrA1,dfrA5,dfrA7/17,dfrA12,dfr85)resistance and drug emux genes(tehA,emrB,emrD,emrE,smr-2,mdfA)were analyzed by PCR and verified by DNA sequencing.Results Resistant rates of these MDR-ABAs to chloramphenicol,rifampicin,tetracycline and trimethoprimsulfamethoxazole were 100.0%(62/62),100.0%(62/62),90.3%(56/62)and 82.3%(51/62)respectively,while62 strains(100.0%),46 strains(74.2%),36 strains(58.1%)and 8 strains (12.9%)were detected to carry mafA,tetB,sull and tehA genes,respectively.The lest 13 genes were all negative.tetB,sull,tehA and mafa genes(2 for each)chosen optionally from positive ones were verified by DNA sequencing and BLASTn.and all were identified as the same sequences in GenBank.Conclusions MDR-ABAs show hish resistance to chloramphenicol,tetracycline, rifampiein and trimethoprimsulfamethoxazole.Multi-drug resistant phenotypes of MDR-ABAs may be closely related to mdfA genes harboring in strains.

Objective To analyze molecular evolution of carbapenemase KPC-12 and its binding free energies with β-lactams.Methods Class A beta-lactamases were divided into 2 clusters:those with carbapenemase activities and those without.Minimum Evolution method in MEGA4.1 software was used to analyze molecular evolution of class A beta-lactamases with carbapenemase activity,including KPC-2 to KPC-13,SFC-1,SME-1,IMI-1,NMC-A,and class A beta-lactamases without carbapenemase activity,including TEM-1,SHV-1.Then,tertiary structure of KPC-12 was predicted by homology modeling as reported in SWISS-MODEL database depending on tertiary structure of KPC-2.Moreover,DOCK module in ArgusLab 4.1 software was used to perform molecular docking of KPC-12 to 10 kinds of beta-lactams substrates,and the binding free energies (△ G) were calculated.Results Molecular evolution between KPC-12 and KPC-2 was the closest.The top three decline in binding free energies of β-lactams were penicillin G sodium salt (△G =-8.45149 kcal/mol),ertapenem (△G =-8.36383 kcal/mol) and ampicillin (△G =-8.19326 kcal/mol),while the last two decline in binding free energies of β-lactams were aztreonam (△G =-6.50614 kca]/mol) and clavulanic acid (△G =-6.88533 kcal/mol).Conclusion Carbapenemase KPC-12 has high catalytic activities to penicillin G sodium salt,ertapenem and ampicillin,while has low catalytic activities to aztreonam and clavulanic acid.

Objective To investigate ADC type cephalosporinase genes in muhidrug-resistant strains of Acinetobater baumannii (MDR-ABA). Methods Sixty-two ABA strains were collected during November 2007 to February 2008 from the First Affiliated Hospital, College of Medicine, Zhejiang University. Kirby Bauer disk diffusion method was used to detect the susceptibilities of the strains to antimicrebial agents. PCR and DNA sequencing was performed to analyze the ADC type cephalosporinase gene. Results Sixty-two MDR-ABA strains showed high drag-resistance rate to most antimicrobial agents expect cefoperazone/sulbactam (69.4%), and 36 strains were ACD positive (58.1%). Conclusion MDR-ABA strains in this study are of high drug resistance, which is closely related to ADC type cephalosporinase.

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.

ObjectiveTo investigate correlation between drug-resistance related genes and mobile genetic elements of Klebsiella pneumoniae resistant to β-lactams. Methods Forty-seven strains of multidrug-resistant Klebsiella pneumoniae were collected from 6 hospitals in Hangzhou and Huzhou of Zhejiang province from August 2008 to May 2010.Modified Hodge test was performed to detect phenotypes of carbapenemases.Forty kinds of β-lactamases (class A-D),ompK35,ompK36,and 12 kinds of mobile genetic elements were detected by PCR,and the results were analyzed by index cluster.ResultsThirty-five strains were positive in modified Hodge test,and 5 kinds of β-lactamases gene ( including KPC-2-like,GenBank:HQ258934) and 9 kinds of mobile genetic elements were detected.Mutations were observed in ompK35 and ompK36 when compared with sensitive strains.Index cluster analysis showed that correlation existed between KPC-2,KPC-2-like and ISKpn6,between TEM-1 and ISEcpl,IS26,int Ⅰ 1,trbC,IS903,and between CMY-2,OXA-30,DHA-1 and tnpU,tnp513,trbC.ConclusionsFive kinds of β-1actamases genes,and mutations in ompK35 and ompK36 may be associated with the resistance to β-1actams in multidrug-resistant Klebsiella pneumoniae.