简介：Objective:Toinvestigatetheapplicationofpolymerasechainreaction(PCR)detectionofHaemophilusducreyiinclinicaldiagnosisofchancroid.Methods:Nucleotidesequencesof16srRNAgenespecificforH.dureyiwereusedtodevelopprimersetsforamplificationoftwostrains.TheamplifiedproductsweretestedviaPCRandsequencedbyelectrophoresisina1.5%gel.TheseproductswerecomparedwiththoseofheterogeneousspeciesorrelatedbacteriatotestthespecificityofthePCRassay.PCRamplificationwithdifferentconcentrationsofH.ducreyiwasperformedtotestitssensitivity.Results:PCRamplificationoftwostrainsofH.ducreyiproducedasinglebandofexpected438bplength.ThesequencewasidentifiedwithgenomicDNA.Noneoftheother19referencespeciesamplifiedunderthesameconditionsgavethisresult.ThehighestsensitivityofPCRassayinthepresenttestwas10ng/L.Conclusions:PCRassayfordetectionofH.ducreyiisarapid,specific,andsensitivedetectionmethod.Iflaboratoryconditionsarestrictlycontrolled,PCRassayisapotentiallyusefullaboratorytestforH.ducreyiinfectiondiagnosis.

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简介：Objective:Toevaluatethevalueofligasechainreaction(LCR)inthediagnosisofdiplococcusgonorrhoeaeinurine.Methods:LCRdetectionoftheurineforNeisseriagonorrhoeaeandbacteriacultureofdischargewasper-formedsimultaneouslyto276patientswithurethritisorcervicitisseekingtreatmentinsextransmitteddis-eases(STDs)outpatientclinic.Forspecimenswithdiscordantresults,polymerasechainreactionwasconducted.ThepurposewastodetecttherespectivesensitivityandspecificityofbacteriacultureandLCR.Results:24of276(8.7%)patientshadpositiveLCRresultsand21of276(7.6%)werepositiveforculture.5specimenshaddiscordantresultsfromLCRandbacteriaculture.ThesensitivityandspecificityofLCRinthediagnosisofgonorrhoeaewere92.3%and100%respectively.Conclusion:ThisstudyshowedthatLCRhadahighersensitivityandspecificityforthediagnosisofgonorrhoeaefromurine.

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简介：Objective:Todetectandquantitategenitalherpessimplexvirus(HSV)DNAinspecimensfrom100patientsclinicallydiagnosedwithgenitalherpes.Methods:PolymeraseChainReaction(PCR)andenzyme-linkedimmunosorbentassay(ELISA)wereusedwithastandardcurveofDNAcopiesofHSVasquantitativecontrast.Results:Ninety-threecaseswereconfirmedHSVpositiveand7caseswerefoundtobenegative.Therewere58casesofHSV-2(62.4%)and35casesofHSV-1(37.6%)amongthe93positivecases.ThenumberofDNAplasmidsrangedfrom115to1.1×l0^5per250pLamongthe93positivesamples(mean=7.1×10^4/250μL).ThenumberofHSVDNAplasmidsrangedfrom136to1.1×l0^5copiesper250pL(mean=7.6×10^4)amongthosewithHSV-2,and115to9.4×10^4per250pL(mean=6.3×10^4)amongthosewithHSV-1.Meanwhile10μLofextractedanddissolvedDNArandomlytakenfrom8eachofHSV-2andHSV-1samplesweretested.ThenumberofHSV-2DNAplasmidsrangedfrom35copiesto2.7×10^4(Mean=l.8×10^4)andthenumberofHSV-1DNArangedfrom29to2.5×10^4(Mean=1.6×10^4).Inthe7negativecases,thequantityofHSVplasmidswaszero.Conclusion:ThesensitivityofELISAquantitation(93%)isequaltothatofSouthernblot.ThesensitivityofPCRfordiagnosisis91%,and88%forPCRtyping.

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简介：AspecificcytotoxicagentagainstgastriccancerwasconstructedbycovalentlycouplingthericinAchaintomonoclonalartibody,MGb2.MGb2wasmodifiedbySPDPtointroducethe3-(2-pyridylthio)propionylradicalandthentreatedwithareducedAchaintogiveadisulfidelinkedconjugatethatretainedtheoriginalbindingspecificityoftheantibodymoiety.TheconjugateobtainedretainedtheactivityoftheantibodyandthebiologicalactivityoftheAchainwell.

简介：Objectives:Todevelopamulti-nestedpolymerasechainreactioninanassaytodetectearlyTreponemapallidumandHaemophilusducreyiDNAintheswabsofgenitalulcers.Methods:Fourpairsofouterandinnerprimers,specifictothebasicmembraneproteingeneofTreponemapallidumandtothe16srRNAgeneofHducreyiweresynthesized.Themulti-nestedPCRwasdevelopedandappliedtodetectTreponemapallidumandHaemophilusdicreyiinclinicalswabs.Result:ThetwosamplesofstandardstrainsofHaemophilusducreyiandoneTreponemapallidumwereamplifiedandshowed309-bprRNAgeneofHaemophilusducreyiand506-bpDNAofTreponemapalidum,respectively.Outof51samplesofgenitalulcerdetected,29showedTreponemapallidumpositiveproductandnoHaemophilusducreyiDNAwasfound.Conclusion:Themulti-nestedPCRforTreponemapallidumandHaemophilusducreyicouldbeusefulforearlydetectionanddistinguishingdiagnosisbetweensyphilisandchancroid.

简介：ObjectiveToexplorethevalueofcomputedtomographyvirtualendoscopy(VE)inassessingossicularchaindisruptionintemporalbonefractureandeartraumawithintacttympanum.MethodsHighresolutionspiralcomputerizedtomography(CT)wascompletedin35casesoftemporalbonefractureand5casesoftympanumtrauma,allwithintactorhealedtympanum.Three-dimensionalreconstructionwascompletedus-ingavirtualendoscopysoftware.Audiologicaltestswereconductedinallpatientsandevaluationoffacialnerveinjuryinpatientswithfacialparalysis.Patientswithmildconductivedeafness,ossicularchainsublux-ationonVE,andnofacialparalysisweretreatedconservativelyfor4-12weekswithrepeatedhearingevalu-ation;thosewithfacialparalysisunderwentsurgeryifnorecoveryafter4-8weeksofconservativetreat-ment.Patientswithmoderatetosevereconductivehearinglossormixedhearingloss,incuslongprocessfractureordislocationonVEandfacialparalysis,underwentossicularchainreconstructionandfacialnervedecompressionafterconservativetreatmentfor4-8weeks,orexploratorytympanotomyonlyifnofacialpa-ralysis.VE,audiologicaltestsandfacialnervefunctiontestswererepeatedin3-6monthsaftersurgery.Re-sultsOfthe6caseswithmildconductivehearingloss,ossicularchainsubluxationandnofacialparalysis,3recoveredtonormalhearingspontaneouslyand3showednosignificantimprovement,after4-12weeksofconservativetreatment.Afterconservativetreatmentfor4-8weeks,3ofthe12caseswithmildconductivedeafness,ossicularchaindislocationonVEandfacialparalysisrecoveredtonormalhearingandHouse-Brackmann(HB)gradeIfacialfunctionfromHBgradeII,4showedfacialfunctionrecoverytoHBgradeI(n=2)orII(n=2)fromHBgradeIIIbutnohearingrecovery,and5gainednorecoveryandwentontoreceiveexploratorytympanotomyandfacialnervedecompression.The11caseswithmoderatetosevereconductivedeafness,incuslongprocessfr

简介：Inflammationafterstrokeisthemaincauseofcerebralischemia/reperfusioninjury.Cascadingeventsafterinjurycanleadtocelldeath.Heatshockprotein70andotherendogenousinjury-signalingmoleculesarereleasedbydamagedcells,whichcanleadtosystemicstressreactions.Protectingthebrainthroughrepairbeginswiththestress-injury-repairsignalingchain.Thisstudyaimedtoverifywhetheracupunctureactsthroughthischaintofacilitateeffectivetreatmentofischemicstroke.Ratmodelsofcerebralischemia/reperfusioninjurywereestablishedbyZeaLonga'smethod,andinjurysiteswereidentifiedbyassessingneurologicalfunction,2,3,5-triphenyltetrazoliumchloridestaining,andhematoxylin-eosinstaining.ElectroacupunctureatacupointsBaihui(DU20)andZusanli(ST36)wasperformedinthemodelratswithdilatationalwaves,deliveredfor20minutesadayat2–100Hzandanamplitudeof2mA.Weanalyzedthebloodserumfromtheratsandfoundthatinflammatorycytokinesaffectedthelevelsofadrenotrophinandheatshockprotein70,eachofwhichfollowedasimilarbimodalcurve.Specifically,electroacupunctureloweredthepeaklevelsofadrenocorticotrophichormoneandheatshockprotein70.Thus,electroacupuncturewasabletoinhibitexcessivestress,reduceinflammation,andpromotetherepairofneurons,whichfacilitatedhealingofischemicstroke.

简介：ObjectiveToexplorethevalueofcomputedtomographyvirtualendoscopy(VE)inassessingossicularchaindisruptionintemporalbonefractureandeartraumawithintacttympanum.MethodsHighresolutionspiralcomputerizedtomography(CT)wascompletedin35casesoftemporalbonefractureand5casesoftympanumtrauma,allwithintactorhealedtympanum.Three-dimensionalreconstructionwascompletedusingavirtualendoscopysoftware.Audiologicaltestswereconductedinallpatientsandevaluationoffacialnerveinjuryinpatientswithfacialparalysis.Patientswithmildconductivedeafness,ossicularchainsubluxationonVE,andnofacialparalysisweretreatedconservativelyfor4-12weekswithrepeatedhearingevaluation;thosewithfacialparalysisunderwentsurgeryifnorecoveryafter4-8weeksofconservativetreatment.Patientswithmoderatetosevereconductivehearinglossormixedhearingloss,incuslongprocessfractureordislocationonVEandfacialparalysis,underwentossicularchainreconstructionandfacialnervedecompressionafterconservativetreatmentfor4-8weeks,orexploratorytympanotomyonlyifnofacialparalysis.VE,audiologicaltestsandfacialnervefunctiontestswererepeatedin3-6monthsaftersurgery.ResultsOfthe6caseswithmildconductivehearingloss,ossicularchainsubluxationandnofacialparalysis,3recoveredtonormalhearingspontaneouslyand3showednosignificantimprovement,after4-12weeksofconservativetreatment.Afterconservativetreatmentfor4-8weeks,3ofthe12caseswithmildconductivedeafness,ossicularchaindislocationonVEandfacialparalysisrecoveredtonormalhearingandHouseBrackmann(HB)gradeIfacialfunctionfromHBgradeII,4showedfacialfunctionrecoverytoHBgradeI(n=2)orII(n=2)fromHBgradeIIIbutnohearingrecovery,and5gainednorecoveryandwentontoreceiveexploratorytympanotomyandfacialnervedecompression.The11caseswithmoderatetosevereconductivedeafness,incuslongprocessfractureordis

简介：Objectives:Toevaluatetheefficacyofnestedpolymerasechainreaction(PCR)withfirstvoidurine(FVU)forthediagnosisofMycoplasmahominisinmalepatients.Methods:MatchedFVUspecimensandurethralswabswerecollectedfrom194malepatientswithNongonococcalUrethritisandtestedbynestedPCRandcellculture.Cellculturewasusedasagoldstandardforevaluatingotherassaytechniques.Results:ForFVUnestedPCRassayandFVUcellculture,ourresultsshowedthatthesensitivitywas100%and93.3%;specificitywas97.0%and98.2%;positivepredictivevalue(PPV)was85.7%and90.3%,negativepredictivevalue(NPV)was100%and98.8%,respectively.Thetotalconsistencybetweenthetwotechniqueswas97.4%.Conclusions:ForthediagnosisofMycoplasmahominisinmen,nestedPCRdetectingFVUisahighlysensitiveandspecificmethod.FirstvoidurinecanreplaceswabcultureorPCRintermsofacceptabilityandfeasibility.

简介：决定区域3的补充(CDR3)的分析一些由immunoscopespectratyping技术的T淋巴细胞受体(TCR)成功地被使用了在自体免疫的疾病和感染疾病调查TCR的差异。在这研究，我们由immunoscopespectratyping技术在四个正常志愿者的人的外部血淋巴细胞为所有32个TCRAV基因家庭调查了CDR3长度分发的模式。PCR产品在1.5%agarose胶化电气泳动上展出了一个卑微的乐队，这被发现。每个TCRAV家庭在6%上展出了超过8个乐队定序胶化电气泳动。所有TCRAV家庭的CDR3spectratyping与不同CDR3长度显示出标准Gaussian分布，并且CDR3的表示频率在基因家庭之中是类似的。大多数在TCRAV家庭的CDR3在框架重新结合。然而，一些CDR3显示出外面框架基因重新整理。另外，我们发现在一些TCRAV家庭，在最长的CDR3和最短的CDR3之间有18氨基酸差异。这些结果可能是有用的进一步在健康人和疾病状态学习人的TCR基因，TCRCDR3基因全部剧目，和全部剧目飘移的再结合机制。

简介：AIMToexploretheprotectiveeffectsandunderlyingmechanismsoftotalpolysaccharidesoftheSijunzidecoction(TPSJ)ontheepithelialbarriersinvitro.METHODSCaco-2cellmonolayersweretreatedwithorwithoutTPSJinthepresenceorabsenceofTNF-α,andparacellularpermeabilityandtransepithelialelectricalresistance(TEER)weremeasuredtoevaluatetheepithelialbarrierfunction.Immunofluorescenceandwesternblottingwererespectivelyusedtoevaluatethedistributionandexpressionofthetightjunctionproteinsclaudin1,claudin2,zo3,andoccludininCaco-2cells.Westernblottingwasalsousedtoevaluatethecellularexpressionofmyosinlightchain(MLC),phosphorylatedMLC(pMLC),MLCkinase(MLCK),andnuclearfactor(NF)-κBp65.RESULTSTPSJpromotedtheproliferationofCaco-2cellsandinhibitedTNF-α-inducedsecretionofpro-inflammatorycytokines.Furthermore,TPSJsignificantlyamelioratedboththereductionofTEERandtheincreasedparacellularpermeabilityobservedintumornecrosisfactor(TNF)-α-damagedCaco-2monolayers.Furthermore,TPSJremarkablyattenuatedTNF-α-inducedmorphologicalchanges,downregulatedtheexpressionofclaudin1,claudin2,zo3,andoccludin,andmarkedlysuppressedTNF-α-mediatedupregulationofp-MLCandMLCKexpression.Finally,TPSJinhibitedtheactivationandexpressionofNF-κBp65.CONCLUSIONOurresultsdemonstratethatTPSJalleviatestheTNF-α-inducedimpairmentoftheintestinalepithelialcellbarrierfunctionbysuppressingNF-κBp65-mediatedphosphorylationofMLCKandMLC.

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简介：Objective:ToevaluatetheclinicalapplicationofmultiplexPCRinthedetectionofTreponemapallidum,Herpessimplexvirus(HSV),andHaemophilusducreyi.Method:ThreestandardstrainswereusedtosetupamultiplexPCR(MPCR)fordetectingsyphilis,herpesgenitalis,andchancroidsimultaneously.Samplesfrom122patientswithgenitalulcerdisease(GUD)weresubjectedtoMPCRandtheresultswerecomparedwiththeseofdark-fiddmicroscopyandTPserology,HSVanligenELISA,andH.ducreyiculture,Result:Inthe122patientswithGUD,MPCRidentified34casesofT.palliduminfection,40casesofHSVinfection,and2casesofmixedinfectionofT.pallidumandherpes.NopositiveresultsofH.ducreyiwerefound.ThesensitivityofMPCRtoT.pallidumandherpeswas100%and93.3%,respectivdy.Thesensitivitiesofdark-fieldmicroscopyandTPserology,HSVantigenELISA,andH.ducreyiculturewas35.3%,50%and100%,respectively.Conclusion:MPCRshowedarelativelyhighersensitivityforT.pallidumascomparedwiththeroutinetechniques.AlthoughitssensitivityforHSVwasnotasgoodasthatofantigenELISA,italsoyieldedahighdetectionrate.MPCRcandetectmorethanonepathogen.Itissimple,quick,sensitive,andsuitableforclinicaluseorepidemiologicalinvestigation.

简介：Objective:TostudytheprevalenceofTrichomonasvaginalis(TV)infectioninChinesemalepatientswithnongonococcalurethritis(NGU),toevaluatethesensitivityandspecificityofurine-basedandurethralswabpolymerasechainreaction(PCR)detection,tosetupamethodfornon-invasivedetectionofmaleTVinfection.Method:OnehundredandfivemaleNGUpatientswereselectedfromaBeijingSTDclinic.Twourethralswabswereobtainedfromeachpatient,onefortheInPouchTVculturesystemandtheotherforPCR.Inaddition,onefirstvoidurinespecimenwascollectedforPCRdetection.Culturewasconsideredthe“goldstandard”.Thesensitivity,specificity,positivepredictivevalue(PPV)andnegativepredictivevalue(NPV)ofthetwoPCRdetectionswerecomparedtocultureresults.Results:Theprevalenceofurine-basedPCRandurethralswabPCRdetectionwas3.81%(4/105)and4.76%(5/105)respectively.Comparedtoculture,thesensitivity,specificity,PPVandNPVwere80%,100%,100%and99%forurine-basedPCRand80%,99%,80%and99%forurethralswabPCR.Conclusion:TVisoneoftheetiologicalagentsinmaleNGU,witha4.76%prevalenceofinfectioninourstudy.TheurinebasedPCRdetectionhashighersensitivityandspecificityandprovidesanoninvasivemethodmorefeasibleinpractice.

简介：AbstractBackground:Rapid and accurate detection of drug resistance in Mycobacterium tuberculosis is critical for effective control of tuberculosis (TB). Herein, we established a novel, low cost strategy having high accuracy and speed for the detection of M. tuberculosis drug resistance, using gene splicing by overlap extension PCR (SOE PCR).Methods:The SOE PCR assay and Sanger sequencing are designed and constructed to detect mutations of rpoB, embB, katG, and inhA promoter, which have been considered as the major contributors to rifampicin (RFP), isoniazid (INH), and ethambutol (EMB) resistance in M. tuberculosis. One hundred and eight M. tuberculosis isolates came from mycobacterial cultures of TB cases at Chongqing Public Health Medical Center in China from December 2018 to April 2019, of which 56 isolates were tested with the GeneXpert MTB/RIF assay. Performance evaluation of the SOE PCR technique was compared with traditional mycobacterial culture and drug susceptibility testing (DST) or GeneXpert MTB/RIF among these isolates. Kappa identity test was used to analyze the consistency of the different diagnostic methods.Results:We found that the mutations of S531L, S315T and M306V were most prevalent for RFP, INH and EMB resistance, respectively, in the 108 M. tuberculosis isolates. Compared with phenotypic DST, the sensitivity and specificity of the SOE PCR assay for resistance detection were 100.00% and 88.00% for RFP, 94.64% and 94.23% for INH, and 68.97% and 79.75% for EMB, respectively. Compared with the GeneXpert MTB/RIF, the SOE PCR method was completely consistent with results of the GeneXpert MTB/RIF, with a concordance of 100% for resistance to RFP.Conclusions:In present study, a novel SOE PCR diagnostic method was successfully developed for the accurate detection of M. tuberculosis drug resistance. Our results using this method have a high consistency with that of traditional phenotypic DST or GeneXpert MTB/RIF, and SOE PCR testing in clinical isolates can also be conducted rapidly and simultaneously for detection of drug resistance to RFP, EMB, and INH.

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