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Objective To investigate the amino acid patterns in penicillin-binding protein 2(PBP2)in Neisseria gonorrhoeae isolates with reduced susceptibility to ceftriaxonc.and the relationship between the amino acid patterns and reduced ceftriaxone susceptibility.Methods DNA was extracted from 13 clinical isolates of N.gonorrhoeae.including 11 strains with decreased susceptibility to ceftriaxone and 2 sensitive isolates.The full-length penA gene encoding the penicillin-binding protein 2 was amplified and sequenced.BLASTn and BLASTx programs were used to assess the insertion and substitution patterns of nucleotides in penA gene and of amino acids in PBP2,respectively.Results BLASTn analysis revealed insertion or substitution of 18-38 nucleotides in the penA gene of gonococcal isolates with reduced ceftriaxone susceptibility.As shown by BLASTX analysis.there were five patterns of amino acid substitution or insertion in PBP2 of the 11 isolates with reduced ceftriaxone susceptibility.However.mosaic structure of PBP2 was not found in any of these isolates.Conclusion Mosaic PBP2 seems not to be the major factor contributing to the decrease in susceptibility of N.gonorrhoeae to ceftriaxone.
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Objectives To investigate the infection and colonization of Mycoplasma genitalium and Ureaplasma urealyticum in different male populations, to explore the association of M. genitalium and U. urealyticum with nongonococcal urethritis (NGU) respectively. Methods A case-controlled, cross sectional study of four different male populations was performed, namely: NGU patients (G1), non-NGU subjects attending STD clinic (G2), men who had sex with men participating in a health education program (G3), and healthy volunteers (G4). Nested PCR and culture were used to detect U. urealyticum. Nested PCR and PCR product sequencing were applied to detect M. genitalium. Results The prevalence rates of M. genitalium in the four study populations were 25.0%(25/100), 6.4%(6/94), 5.5%(6/110) and 0% respectively. Significant difference was found between each two groups except G2～G3 with a p value of 0.80. By multivariate regression analysis, controlling for the age of first sex, new sexual partners, urethritis and condom use in the previous 3 months, M. genitalium was only associated with urethritis (P= 0.004, OR = 6.754, 95% CI 1.833～24.893). The direct sequencing of PCR products showed gene mutations, in comparison with the reference sequence in GenBank, in 3 samples. The prevalence rates of U. urealyticum by PCR in 4 groups were 40.0%, 44.7%, 22.7% and 46.9% respectively, and there was no significant difference between G1～G2, G1～G4 or G2～G4 with a p value of 0.419, 0.325, 0.868 respectively, but the prevalence rate of U. urealyticum in G3 was significantly lower than that in other groups. Conclusions M. genitalium is strongly associated with NGU and the prevalence rate is significantly higher in groups with high risk sexual behaviors than that in general population. There is no association between the colonization of U. urealyticum and NGU.
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Objective To investigate the expression and localization of platelet-derived growth factor receptor-?and-?(PDGFR-?and-?)in keloid tissues and keloid-derived fibroblasts and to explore the role of PDGFR in the pathogenesis of keloid.Methods The distribution of PDGFR was determined by im-munohistochemistry technique in15keloid lesions and10normal skin tissues.The expression of PDGFR protein in vitro was identified by Western blotting analysis.Results Strong staining of PDGFR-?was locat-ed in keloid tissue.However,the expression of PDGFR-?in keloid tissue seemed to be related to the status of clinical proliferation.Strong expression of PDGFR-?was observed in the lesions with a congestive and invasive margin,whereas mild expression of PDDFR-?was found in lesions with a stable margin.Expression of PDGFR-?protein was more abundant than that of PDGFR-?protein in fibroblasts in vitro.Conclusion The elevated levels of both types of PDGFR might facilitate their responses to PDGF in keloid fibroblasts,which seems to play an important role in the formation of keloid.
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Objective To investigate the role of connective tissue growth factor(CTGF)in the pathogenesis of keloid.Methods The expression of CTGF mRNA was detected with semiquantitative RT-PCR technique in keloid tissue and adjacent uninvolved skin from30patients.The expression of CTGF pro-tein was determined by SP immunohistochemistry in5keloid tissues and5normal skin samples.Results Compared with normal controls,the mean levels of CTGF mRNA expression were significantly increased in both keloid lesions and adjacent uninvolved skin(P0.05).In immunohistochemistry study,CTGF protein was strongly expressed in keloid tissue,whereas it was not expressed in normal skin.Transitional phenomenon was observed from strong to weak expression of CTGF protein at the edge of keloid tissue.Conclusion These findings suggest that CTGF may be related to the development of persistent fibrotic tissue formation in keloids.CTGF may be a new potential target for the therapeutic intervention of keloids.
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Objective To investigate the antimicrobial susceptibility of Neisseria gonorrhoeae isolated from 10 cities of China, and to provide reference for the formulation of treatment guideline and making control strategy. Methods Agar dilution was used to detect antimicrobial susceptibility and acidometric method for PPNG testing. Results A total of 3186 gonococcal isolates were tested from 1993 to 1998. The resistant rate for penicillin was 66.70％ . PPNG isolates accounted for 8.14％ . Tetracycline－ resistant isolates accounted for 93.02％ , and high level tetracycline－ resistant isolates (TRNG) accounted for 4.65％ . The resistant rate for ciprofloxacin was also high (34.25％ ). The resistant rates for spectinomycin and ceftriaxone were 0.44％ and 0.57％ respectively. Conclusions The gonococcal isolates are highly resistant to penicillin, tetracycline, and ciprofloxacin, while still sensitive to spectinomycin and ceftriaxone. These results suggest that the present treatment for gonorrhea can still be used.
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ObjectiveTounderstandthetrendsandepidemiologicalcharacteristicsofsexuallytransmitteddiseases(STDs)inChinaandprovidescientificbasisformakingcontrolstrategies.MethodsDuringtheperiodof1991～2001,thecase-reportingdataof8kindsofnotifiableandmonitoringSTDs,collectedfrom31provinces,autonomousregionsandmunicipalities,wereanalyzedwithepidemiologicalmethods.Results①Epidemictrends:Duringthisperiod,theincidenceof8kindsofSTDssteadilyincreasedfrom175528(15.48per100000population)in1991to859040(68.91per100000population)in2000.Theaverageannualgrowthofincidencewas19.30%,witharangeof2.59%～36.88%.However,thereportedcasesin2001were795612withadecreaseof7.38%comparedwiththosein2000,anditwasthefirstdecreasesinceSTDcaseswerereportedfrom1987.②Geographicaldistribution:Thehigh-incidenceareasweretheZhujiangRiverDelta,YangtzeRiverDelta,MinjiangRiverValley,NortheasternChina,andBeijing,Tianjin,andChongqingManicipalities,withtheincidencerateofover70～100casesper100000populationafter1997,andtherewereveryhighratesofincidenceover1000casesper100000populationinsomeareas.Thelow-incidenceareaswerenorthChina,partsofCentralChina,NorthwesternChinaandSouthwesternChina,withtheincidenceratesoflowerthan30～50per100000population.③Populationdistri-bution:Themaletofemaleratiodecreasedfrom1.60∶1～1.69∶1intheearly1990sto1.35∶1～1.40∶1inthelate1990s.STDincidencerateswerehighestinthe20～39agegroup,andthereportedSTDcasesofthisagegroupaccountedforover80%oftotalcases.ConclusionSexuallytransmitteddiseasesinChinahavebecomeaseriouspublichealthproblemandtheeffectiveinterventionprogrammesagainstSTDsmustbeimplementedacrossthecountry.
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ObjectiveTodetectM.genitalium(Mg),M.penetrans(Mpe),M.pirum(Mpi),M.fermentans(Mf),Ureaplasmaurealyticum(Uu)andM.hominis(Mh)infectionsinurethra/cervicalcanalandpharynxandexploretheirclinicalsignificance.MethodsCultureandPCRwereperformedin72patientswithNGU/MPCtodetectMg,Mpe,Mpi,Mf,UuandMh.Thesecretionsfromurethra/cervicalcanalandpharynxweretested.ResultsMg,Mpe,Mpi,Mf,UuandMhweredetectedfromgenitalspecimensin23.6%,12.5%,2.8%,0,26.4%and8.5%ofpatients,respectively.Mg,Mpe,Mpi,Mf,UuandMhweredetectedfrompha-ryngealspecimensin24.6%,14.5%,0,0,2.9%and2.9%ofpatients,respectively.Thesamespeciesofmy-coplasmaswerefoundinbothgenitalandpharyngealspecimensin10patients(14.5%).ConclusionsUuandMginfectionsarecommoninpatientswithNGU/MPC.ThenewmycoplasmaspeciesMpeshouldbepaidattentionto.TheresultsindicatethatMgandMpemaybetransmittedbygenital-genitalsexandoral-genitalsex.MfmaybeofnoassociationwithNGU.
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Objective To investigate the predisposing alleles of HLA-DR genes in pemphigus. Methods Polymerase chain reaction specific sequence primers (PCR-SSP) method was applied to type HLA-DR subregion in the patients with pemphigus vulgaris (PV), pemphigus erythematosus (PE) and matched control subjects of Han nationality from Jiangsu and Anhui provinces. Results The results demonstrated that DR4, DRB1*14 (*1401, *1404,*1405)gene frequencies were significantly higher in both PV(Pc
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0 05), except serotype 3 from NGU patients, which was more commonly found in women than in men (P10%, respectively). Conclusion Serotype 4 was the one most frequently found and strongly related with NGU. It is suggested that the transformation from colonization to pathogenetic status be possible for serotype 3 under the influence of host and a certain environmental condition.
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Objectives To test a simple method for genotyping of C.trachomatis isolates and to investigate the clinical significance of the genotypes. Methods A part of the chlamydial genome encoding the major outer membrane protein(omp1) was amplified by polymerase chain reaction (PCR). The products were digested by endonucleases to see the characteristic patterns, after silver staining on 10% polyacrylamide gels. Results The omp1 genes of 15 serovars of C. trachomatis were amplified by PCR,which generated an 871 base pair gene fragment. AluⅠ digestion of the product gave characteristic patterns for the 15 serovars,but group C presented closely similar patterns. A triple digestion with HpaⅡ, followed by HinfⅠ and EcoRⅠ, would allow the differentiation of serovars in group C. Analysis of 74 clinical isolates revealed serovars E, F, D, G as the most prevalent genital serovars in the studied populations. Serovars B, H, J were occasionally identified. A mixed infection with serovars F and D was seen in a clinical sample. No significant relationship was observed between clinical manifestations of urogenital chlamydial infections and serovars,however,serovar D was more often associated with high titer of anti chlamydial antibody than other serovars. Conclusion The omp1 genotyping technique seems to be promising for epidemiological studies.
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Objective To evaluate the ability of restriction endonuclease analysis by pulsed field gel electrophoresis (PFGE REA) and random amplified polymorphic DNA (RAPD) analysis for molecular typing of Neisseria gonorrhoeae isolates. Methods Genomic DNA from 36 Neisseria gonorrhoeae isolates were digested with SpeI and analysed by contour clamped homogeneous electric fields electrophoresis. The 36 isolates were also characterised by RAPD analysis with arbitrary primer OPA 03. Results 12～19 DNA fragments from 5 to 600kb were obtained among the 36 isolates after PFGE REA. The 36 isolates were discriminated into 20 PFGE patterns. Amplification of Neisseria gonorrhoeae DNA with primer OPA 03 produced 15 different DNA fragments (280～1900bp), and 3～9 fragments per strain could be seen. The 36 isolates showed 18 different RAPD patterns. Strains sharing common auxotypes and antibiotic spectrum could be differentiated by PFGE REA and RAPD analysis. General agreement was found between these two techniques. Isolates from different geographic areas and even some isolates in the same area showed considerable amount of DNA polymorphism. In addition, some isolates shared common or very similar patterns were unique to a given geographic area. Conclusion It is concluded that both PFGE REA and RAPD analysis are useful, sensitive molecular techniques for differentiation of clinical isolates of Neisseria gonorrhoeae. They should be helpful in the investigation of strain origin, clonal relation among strains and spread of antibiotic resistance. Compared with PFGE REA, RAPD analysis is faster, relatively simple and more economical.
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Objective To monitor antibiotic resistance of Neisseria gonorrhoeae in Nanjing City based on analysis of the results from 1999 to 2002. Methods The production of ?-lactamase was determined by paper acidometric method. Minimum inhibitory concentrations (MICs) of penicillin, ceftriaxone, tetracycline, ciprofloxacin and spectinomycin were determined by an agar plate dilution method. Results A total of 417 clinical isolates of Neisseria gonorrhoeae were examined. During the period of 1999-2002, positive rate of PPNG rapidly increased from 8.0% to 31.31% (P