简介：TheaimofthisstudyistoinvestigatecyclinE,pl6inkdaandki67aspossiblediagnosticbiomarkersforcervicalpreneoplasiausingcervicalexfoliated-cellspecimens,andevaluatethesignificanceforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.TheexpressionofcyclinE,pl6inkdaandki67wasexaminatedin78cervicalexfoliatedepithelialspecimensdiagnosedasatypicalsquamouscellsofundeterminedsignificance(ASCUS)(12cases),cervicalintraepithelialneoplasia(CIN)oftype1(17cases),CIN2_3(38cases)andinvasivecarcinoma(11cases)usingimmunohistochemicalanalysis,andsimultaneously,theDNAstatusofhumanpapillomavims(HPY)type16/18wasdetectedbypolymerasechainreaction(PCR)usingtypespecificprimers,cyclinE,pl6inkdaandki67werealloverexpressedinCINsandinvasivecarcinoma,comparedwithlittleexpressioninASCUS(P<0.005).OverexpressionofcyclinEwasobservedinCIN1(94.1%,X^2=21.16,P<0.01),andp16inkdaandki67wereoverexpressedininvasivecarcinoma(100%and90.9%respectively).Thedegreeofpl6inkdaandki67expressioncorrelatedwellwiththatofepitheliallesions(P<0.005).HPV16/18infectionwasassessedinC1Nsandinvasivecarcinomasamples,andrevealedasignificantrelationshipwiththedegreeofcervicalepitheliallession.Theexpressionlevelofpl6inkdaandki67seemedmorecloselyassociatedwithHPVI6infectionthanthatofcyclinE(rs=1.0vsrs=0.4).Only1caseinCINIanddcasesinCIN2-3ofHPV18positivesamplesweredetected.Thereforenostatisticalsignificancewasfoundbystatisticalanalysis.OverexpressionofcyclinE,pl6inkdaandki67inCINsandinvasivecarcinomacellsdemonstratesthepotentialuseofcyclinE,pl6inkdaandki67asdiagnosticbiomarkersforHPV-relatedcervicalneoplasticlesions.Inaddition,thistechniquecanbeusedforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.

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简介：Toestablishasensitiveandspecificmethodforseroepidermiologicaldetectionofhumanher-pesvirus8(HHV-8)infection,threepotentantigenicproteinsencodedbyopenreadingframes(ORFs)K8.1,65and73CingenomeofHHV-8wereproducedasglutathioneS-transferasefusionproteinintheprokaryoticexpressionsystemandwasusedasantigenfortesting.Therecombinantfusionproteinex-pressedintheprokaryoticexpressionvectorE.coliBL21waspurifiedbyglutathioneSepharose4Baffin-itychromatographyandwasquantitatedwithSDS-PAGE.Allthese3fusionproteinsproducedinthepro-karyoticexpressionsystemshowedgoodimmunogenicityasdemonstratedbyWesternblottingandcouldberecognizedbymixedseraofpatientswithKaposi′ssarcoma(KS).Theimmuno-reactivitiesofthesingleorcompoundfusionproteinweredeterminedbymeansofELISAandcomparedwiththetraditionalimmu-nofluorescenceassay(IFA)todeterminetheirsensitivityandspecificityofthetest.Itwasdemonstratedthatthesensitivityofmixed-antigenELISAmethodwassignificantlyhigherthanthatofIFA(81.8%vs34.4%),whilethespecificityoftheformerwasdemonstratedtobe97.9%.Thecoincidenceofthede-tectionratebetweenthesetwomethodswasconsiderablyhigh,approachingupto90.0%.TheseresultssuggestthatthemixedantigenELISAassayappearstobeasensitiveandspecificmethodforsero-epide-miologicaldetectionofhumanherpesvirus8infection.

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简介：【摘要】目的：探讨 8S管理模式在消毒供应中心外来器械全流程管理中的应用效果。方法：本课题选取 2019.01到 2019.11时段内收取的外来器械共 600件，以随机抓阄法纳入参照组（ 300例）、管理组（ 300例）。即参照组为传统器械管理，管理组为器械 8S管理模式，比较器械质量合格率、消毒供应中心员工自我评价和满意度评分。结果：参照组器械质量合格率较低于管理组，即 91.67%＜ 96.67%，数据间比较有意义（ P＜ 0.05）。管理组消毒供应中心员工自我评价、满意度评分均优于参照组，数据间比较有意义（ P＜ 0.05）。结论：在消毒供应中心外来器械全流程管理过程中， 8S管理模式的选择不仅可提高器械质量合格率，还可增强消毒供应中心员工自我评价和满意度，应引起重视。

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简介：Chlamydiatrachomatisoutermembraneprotein2(Ctomp2)isamajorimmunogeninchlamydialinfectionsandahighlygenus-conservedstructuralproteinofallChlamydiaspecies.Topurifytheproteinandtopreparemonoclonalantibodies(mAbs)againstit,therecombinantproteinwasinducedbyIPTG,whichwasconfirmedbySDS-PAGEandpurifiedbymeansofaNi2+-chargedresincolumn.ThedenaturedproteinwasrefoldedintheGSH-GSSHbuffergraduallyandidentifiedbyWesternblotting.ThentheBALB/cmicewereimmunizedwiththerecombinantproteintopreparethemAbagainstCtomp2.TheobtainedmAbswerecharacterized.GenitalspecimensweretestedwithindirectELISAmostlymadeofthemAbandcellculturein84patientswithgenitalsymptoms.Theresultsshowedthathigh-levelexpressionoftherecombinantproteinwasachieved,whichexistedasinclusionbodyandamountedto38%oftotalbacteriumprotein.AmAbagainstCtomp2wasobtained.ItbelongstoIgG2b.Thetiterswereashighas1:40000.TheWesternblottingshowedthatthemAbonlyreactedwiththerecombinantprotein.IthadnocrossingreactionsagainstE.coli,N.gonorhoea,M.hominis,U.urealyticumandM.penetrans.Ithadhighspecifity.Incomparisonwithgoldstandardtest-cellculture,thesensitivities,specificities,positivepredictivevaluesandnegativepredictivevaluesofindirectELISAwere95.24%,100%,100%and98.44%,respectively.Theabove-mentionedresearchworkcontributednotonlytothefurtherstudyofthestructureandfunctionofthisprotein,butalsototheestablishmentofthemethodforitsclinicalapplication,forithadnotbeenreportedbefore.

简介：ToinvestigatetheexpressionlevelsofthreeDsbproteingenes,dsbB,dsbDanddsbG,atdifferenttimepointspostC.trachomatisinfection,mousefibroblast12cellswerechosentobeinfectedwithC.trachomatisserovarFstrainF/IC-Cal-13.C.trachomatiselementarybody(EB)-infected12cellswereharvestedimmediatelyafterEBattachmentontothecellsandevery4hourspostinfection(hpi)till44hpifortotalRNApreparation.RT-PCRassayswerethenemployedtoamplifycDNAwithprimerpairswhicharespecifictoC.trachomatisdsbgenesdsbB,dsbD,dsbGandtufArespectively.Therela-tiveexpressionlevelsofDsbproteingeneswereevaluatedascDNAratiosofgenedsbtogenetufA.OurresultsshowedthatthetranscriptionofdsbGstartedfrom12hpiandgraduallyincreasedtill44hpi.ThetranscriptionofdsbBanddsbDweredetectedat16hpiandreachedtheirpeaksat28hpiand24-28hpi,respectively.Moreover,therewasobvioustranscriptionofdsbBatthelaterstage(44hpi),butnonefordsbDatthistimepoint.WecametotheconclusionthattheexpressionlevelsofthethreeDsbproteingenesaredifferentduringthedevelopmentalcycleofC.trachomatist.Theymayplayaroleinmid-to-latestageofthedevelopmentalcycleofC.trachomatis.

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简介：Thehousedustmites(Dermatophagoidesfarinae,Derf)arethemajorsourceofaeroallergensimplicatedintheexpressionofatopicdisorders,includingasthma,allergicrhinitisandatopicdermatitis.Inparticular,strongcircumstantialevidencesuggeststhathousedustmiteantigensareimportantprecipitatingfactorsofasthma.Manyhousedustmiteallergensareproteasesthatcanelicitairwayinflammationbystimulatingthereleaseofcytokinesfrombronchialepithelialcells.ToinvestigatewhetherDerfallergenproteasesinducedcytokineproductionfromtheepithelialcelllineBEAS-2B,BEAS-2Bcellswereculturedwith4differentconcentrationsofDerf(0.02,0.2,2,20μg/ml)for24-96h,afterwhichsupernatantswereassayedforinterleukin(IL)-6andIL-8withELISA.Reversetranscription-PCRwasalsoperformed.ThecellsheetswereintactthroughouttheobservationincontrolgroupwithoutanyexposuretoDerfantigen.IntheexperimentalgroupscellstreatedwithDerfallergenshowedchangesintheanchoragestatusofthemonolayer.Therewasasignificantincreaseinthelevelofcytokineproductioncomparedwiththeuntreatedsample.ThereleaseofIL-6andIL-8increasedinaconcentration-dependentmanner(P<0.05,respectively)withtheadditionofincreasingdosageofDerftothecellsheets.LevelsofIL-6andIL-8begantoriseat24hand48hafterallergenexposure,andtheyincreasedsignificantlyinthesupematantsat72hand96h.AtthesametimetheconcentrationdependenceofinductionofIL-6andIL-8expressionaswellasanincreaseintheexpressionofIL-6andIL-8mRNAmanifestedevidently.HDM-inducedairwayinflammationmayincludeDerf-mediatedreleaseofinflammatorymediators,andtheproteolyticactivityofanallergenmaystimulatethereleaseofproinflammatorycytokinesfromhumanbronchialepithelium.ItissuggestedthatIL-6andIL-8productionbybronchialepithelialcellsmayplayaroleinthepathogenesisofallergicasthma.

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简介：ToconstructandexpressthefusionproteinStx2B-IntiminC300ofEHEC0157:H7,andtofurtherinvestigateitsimmunoprophyiacticpotential,thegeneofStx2B(stx2b)fromEHEC0157:H7chromosomewasclonedintopMD18-Tvector.Thereafter,theamplifiedgenewasclonedintoprokaryoticexpressionplasmidpET-28a(+)-eaeC300,whichwasconstructedpreviously.TherecombinantpasmidpET-28a(+)-stx2b-eaeC300wastransformedintoE.coliBL21(DE3).Afterinducement,theproteinStx2B-IntiminC300wassuccessfullyexpressedandanalyzedwithsodiumdodecylsulfatepolyacrylamidegelelectrophoresis(SDS-PAGE),WesternblottingandN-terminalaminoacidresidualsequencing.Toevaluateitsimmunoprophyiacticpotential,itwasprimarilypurifiedbyion-exchangechromatographyandinjectedinto30BALB/cmicewithAl(OH)3inthesubscapularregion.Tendaysafterthelastboostervaccination,20micewereattackedwithEHEC0157:H7lysateandtheprotectiveefficacywasobserved.Inthepresentstudy,thegeneofStx2B-IntiminC300wassuccessfullyclonedintopET-28a(+)vector.TheresultsofSDS-PAGEandWesternblottingassayshowedthatthefusionproteinwassuccessfullyexpressedintheinclusionbodyform,accountingfor25%oftotalexpressionproducts,anditsmolecularweightwasabout43kDa.TheresultoftheN-terminalaminoacidresidualsequencingshowedthatitwasidenticaltothatofthemoleculardesigned.Thepuritywasabout75%afterprimarypurification.AnimaltestsrevealedthatthefusionproteinStx2B-IntiminC300haselicitedhightiterofprotectiveantibodyrelatively.TheseresultsdemonstratethatthefusionproteinStx2B-IntiminC300issuccessfullyexpressedinprokaryoticexpressionsystemandshowscertainimmunoprophyiacticpotential.

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