简介:Objective:ToobtainrecombinantTreponemapallidumsubsp,pallidum(TP17KD)lipoproteininlargequantitiesbyamplificationandtofurtherpurifyantigensforlaboratorydiagnosisofsyphilisanddevelopmentofasyphilisvaccine.Method:TheTppl7lipoproteingenewasamplifiedfromtheTP(strainNichols),andthenitwasrecombinatedintoaplasmidpMAL-2candclonedwithinE.coli12-TB1.ThehostbacteriacontainingrecombinantplasmidswereinducedwithIPTG.TheTpp17KDlipoproteingenewasamplifiedbyus-ingPCRandpositivecloneswerescreenedwithdoubledigestionandPCR.RecombinantplasmidsweretransformedintoE.coliandtheE.colicarryingrecombinantplasmidswereinduced.TheexpressionofTP17KDwasdetectedbysodiumdedecylsulfate-polyacrylamidegelelectrophoresis(SDS-PAGE)andimmunoblot.Results:GelstainingwithCoomassieblueG-250showedthattheinducedE.colicarryingrecombinantplasmidcouldproduce60KDfusionproteinathighlevels.Gelscanningshowedthat17KDproteinexpressioninE.coliaccountedfor10%oftotalcellularprotein.Therecombinantproteinantigenreactedwiththeseraofsyphilispatients.Conclusion:Ourstudylaysacornerstonefordevelopingnewtechniquesoflaboratorydiagnosisforsyphilisandnewvaccines.Preliminaryclinicalapplicationshowedthatthefusionproteincouldbeusedforthediagnosisofsyphilis.
简介:Objective:Todevelopasensitive,specificandsimplemethodfordetectionofextremelylownumbersofT.palliduminclinicalspecimens,asasignificantadditiontotheserologictestsforsyphilisdiagnosis.Methods:Double-tubenestedPCR(DN-PCR)andsingle-tubenestedPCR(SN-PCR)assayswereperformedtoamplifyspecificfragmentsoftheDNApolymeraseIgene(polA)ofT.pallidum.SensitivityandspecificityofthetwoPCRassaysweretested.EightysixwholebloodspecimensfrompersonswithsuspectedsyphilisweredetectedbythetwonestedPCRmethods.TheTPPAtestwasusedasacomparisonfordetectingsyphilisinserafromcorrespondingpatients.Results:OnlyspecificampliconscouldbeobtainedduringamplificationoftheT.pallidumpolAgeneandthedetectionlimitwasapproximately1organismwhenanalyzedongelbythetwoPCRmethods.Of86clinicalspecimens,62werepositivebyTPPA.Ofthese,54and51werepositivebytheDN-PCRandSN-PCR,respectively,whichdoesnotrepresentastatisticallysignificantdifferencebetweenthetwoPCRtests.Of24TPPA-negativespecimens,5werepositivebybothDN-PCRassayandSN-PCRassay.Conclusion:TheSN-polAPCRmethodisextremelysensitive,specificandeasytoperformfordetectinglownumbersofT.palliduminclinicalbloodspecimensasacomplementarytoserologyforsyphilisdiagnosis.
简介:Objectives:TodevelopamethodofsimultaneousPCRdetectionofHaemophilusducreyi,Treponemapallidum,andHerpesSimplexVirusTypes1and2fromgenitalulcersamongpatientsattendingSTDclinicsinGuangzhou,China;andevaluatetheclinicalapplicationofmultiplexPCR(M-PCR)assayfordiagnosingtheetiologyofgenitalulcerdiseases(GUD).Methods:244patientswithagenitalulcerwereevaluated.ClinicaletiologyofGUDwasbasedonphysicalappearanceandmicrobiologicevaluationsthatincludeddarkfieldmicroscopyexamination(D-F)andserologytestforsyphilis(STS).SwabsofeachgenitalulcerweretestedforHSVantigenbyenzymeimmunoassay(EIA)andprocessedinanM-PCRassayforsimultaneousdetectionofT.pallidum,HSVandH.ducreyi.Results:ThestandardstrainsofT.pallidum,HSVandH.ducreyiwereamplifiedbyM-PCR,producingamplifiedproductsof260bp,432bp,170bp,respectively.ThesensitivityofM-PCRis102pgDNA.M-PCRassayforT.pallidum,HSVandH.ducreyishowedgoodagreementwhencomparedwithD-FdetectionforT.pallidum,STS,H.ducreyicultureandEIAforHSVantigen(Kappascoresare0.774,0.704,0.793,0.756,respectively).Conclusions:TheM-PCRisaconvenient,accurateandreliableassayforthedetectionofT.pallidum,HSVandH.ducreyifromgenitalulcers,andcanbeusedasamethodofdiagnosingtheetiologyofGUD.
简介:目的:探讨孕前规范驱梅治疗的梅毒血清固定患者在孕期是否需要驱梅治疗。方法:回顾性分析我院2010~2013年无系统感染的梅毒血清固定育龄妇女的临床资料,按妊娠期是否接受驱梅治疗分为治疗组(A组)和未治疗组(B组),比较其妊娠结局的差异。结果:共收集了68例患者,A组46例,B组22例,两组患者足月产分别有44例(95.65%)和21例(95.45%),早产分别有2例(4.35%)和1例(4.55%),低体重儿两组各有1例(2.2%、4.5%),两组均无先天梅毒儿出生。两组在足月产、早产、低体重儿、先天梅毒儿发生率上,差异均无统计学意义(P值均〉0.05)。结论:孕前经过规范驱梅治疗的育龄妇女,如果没有合并系统感染,妊娠期在密切随访下可能无需予驱梅治疗。
简介:本研究中,作者调查了以慢性炎症和内皮功能障碍为特征的白塞病(BD)患者外周组织胰岛素抵抗的情况。14例BD患者和15例健康对照者被纳入本研究。采用正常血糖高胰岛素钳夹技术研究胰岛素抵抗的情况。与健康对照者相比,BD患者胰岛素抵抗的发生率高(P=0.014)。BD患者的胰岛素敏感性(M),即在血糖正常的稳态条件下测量的葡萄糖利用率,与对照组相比显著降低[4.09±0.16mg/(kg·min)vs5.60±0.27mg/(kg·min),P=0.001]。C反应蛋白(CRP)水平而非血沉(ESR),与胰岛素抵抗显著相关(CRP:rs=0.589,P=0.27;ESR:rs=0444,P=0112),未发现M值与ESR或CRP有关系。作者推测BD患者存在外周组织胰岛素抵抗,可能是BD的炎症和内皮功能异常导致了这一系列不同后果。
简介:目的:检测梅毒血清固定患者外周血Th9细胞的表达水平,探讨其与血清固定免疫机制的关系。方法:收集梅毒血清固定患者(15例)、正规驱梅治疗后血清转阴者(8例)及正常对照者(15例)外周血单个核细胞,采用流式细胞仪检测Th9细胞比例。结果:血清固定组Th9细胞比例较血清转阴组和正常对照组降低,差异有统计学意义(Z=-3.49,P〈0.001;Z=-3.40,P=0.001);血清转阴组Th9细胞比例较正常对照组稍高,但差异元统计学意义(Z=-1.03,P=0.301)。结论:Th9细胞可能通过一系列免疫过程参与了血清固定的发生发展,具体机制仍有待进一步明确。
简介:目的对比给予高位隐睾患儿开放隐睾手术、腹腔镜下隐睾下降固定术治疗的临床效果。方法选取2016年1月至2018年1月新疆伊宁市兵团四师医院诊治66例高位隐睾患儿为研究对象。将其分为开放组(n=33)、腹腔镜组(n=33),分别给予开放手术睾丸下降固定术治疗、腹腔镜下隐睾下降固定术治疗,观察患儿睾丸发育状况、并发症发生情况等。结果腹腔镜组术中出血量显著少于开放组,手术时间及术后切口疼痛、首次下床活动、住院时间均显著短于开放组,差异具有统计学意义(t=15.152、9.030、28.614、16.893、10.183,P<0.05);术后6个月,2组患儿月睾丸发育良好率分别为84.85%、75.76%,组间比较差异无统计学意义(χ~2=0.862,P>0.05);术后3、6个月,2组患儿睾丸容积均显著大于术前,差异具有统计学意义(F=7.967、7.996,P<0.05),但组间比较差异无统计学意义(P>0.05);腹腔镜组治疗总有效率为84.85%,相比开放组的92.31%差异无统计学意义(χ~2=0.569,P>0.05);腹腔镜组无患儿出现并发症,总发生率为0,相比开放组的27.27%显著更低,差异具有统计学意义(χ~2=10.421,P<0.05)。结论给予高位隐睾患儿腹腔镜下隐睾下降固定术,总体疗效与开放隐睾手术相当,但并发症更少,安全性更高。
简介:临床资料患者,男,65岁。主因外阴红斑伴痒痛6年,于2014年11月21日就诊。6年前,无明显诱因患者左侧阴囊皮肤出现一绿豆大“疖肿”样皮损并化脓,自行处理并好转后于原部位出现瘙痒性红斑,逐渐向周围扩大,偶有渗出、结痂,时有轻度瘙痒、辣痛感,反复搔抓后红斑处糜烂;于多家医院按湿疹予外用药治疗(具体不详),症状可稍缓解,但皮损从未完全消退,渗出与干涸交替。红斑渐向周围扩大,逐渐累及阴茎及左侧阴囊、阴阜。患者既往体健,无系统性疾病。
简介:报告1例新生儿先天性无汗型外胚叶发育不良。患儿男,14天,出生后皮肤干燥、脱屑,间断发热2周。躯干皮肤病理显示皮肤萎缩变薄,真皮层未见明显的毛囊、汗腺及皮脂腺等皮肤附属器,小血管周围少量淋巴细胞浸润。