简介:【摘要】目的:讨论乙肝病毒性肝炎患者临床医学检验应用探究。方法:选取我院治疗的乙肝病毒性肝炎的患者50例,均实行两对半和HBV-DNA定量检测,回顾性分析检测结果,选取的患者均需要使用两对半临床医学检验,采集患者的清晨空腹状态下静脉血4毫升,将血液放入离心机中,使用每分钟3000转,10分钟后,将血清分离出来,放在4摄氏度的条件下保存。结果:所有患者经临床检测诊断结果均为乙肝病毒性肝炎,其中小三阳患者占18例(36.0%),大三阳患者占13例(26.0%),其它类型占19例(38.0%),三组之间的差异性较为显著(P
简介:Inordertoanalyzethesequencesoftheinternaltranscribedspacer(ITS)includingthe5.8SribosomalDNA(rDNA)ofcommondermatophytes,soastoobtainarapidandaccuratemethodtoidentifythespeciesofdermatophytesandtoestablishthephylogenetictreeofthesespeciestounderstandtheirrelationship,16strainsofdermatophyteswerecollectedandpreliminarilyidentifiedbymorphologicalcharacteristics.GeneralprimersforfungiITS1andITS4wereusedtoamplifytheITSrDNAofeachstrainswithPCR.ThePCRproductsafterpurificationweresequenceddirectlyandwereanalyzedthroughinternet.Intheresults,11strainswereidentifiedbymeansofmorphologicalfeatures,amongwhich5strainswereTrichophyton,5strainswereMicrosporumand1wasEpidermaphyton,whichwasconsistentwiththeresultsbymolecularbiology.Inthe5unidentifiablestrains,1strainwasprovedtobeChrysosporiumbymolecularbiology.Thesestrainsstudiedcouldbedividedinto3differentclassesasindicatedintheanalysisofthephylogenetictreeofthesequencesinITS,whichwerequitedifferentfromthoseofmorphologicalclassification.ItisevidentfromtheaboveobservationsthatthemolecularmethodofanalysisontheITSsequencesisarapid,highlysensitiveandaccurateapproachforthedetectionofdematophytespecies,however,itstillexhibitssomelimitationsneedingthesupplementationwithmorphologicalidentification.
简介:TostudytheexpressioncharacteristicofJapaneseencephalitisvirus(JEV)prMEandEproteinsandtheefficacyofDNAimmunizationbydifferentrecombinantplasmidscontainingJEVprME(2001bp)andE(1500bp)genes,tworecombinants(pJMEandpJE)containingJEVprMEandEgenesfusedwithFLAGwereconstructedandthentransfectedintoHepG2andCOS-1cellsbylipnsomefusion.TheexpressionfeatureofFLAG-prME(about72kDa)andFLAG-E(about54kDa)proteinsintransfectedcellswereanalyzedbyWesternblotandtwoantibodysystems(anti-FLAGandanti-E).BALB/cmicewereimmunizedwith100μgoftwokindsofrecombinantsbyintramuscularinjection,andJEVJaGAr-01strains(10^5PFU/100μl)weregiventoBALB/cmicebyintraperionealinjection3wkaftertwiceDNAimmunizationbyalethalviruschallenge.BALB/cmicewereobservedfor21daysafterchallenge.80%plaquereductionneutralizationtestwasperformedtotitrateneutralizationantibodybeforeandafterviralchallenge.ItwasfoundthattheexpressionofproteinsassociatedwithpJMEandpJEwasdeterminedintransfectedcellswithanti-FLAGandanewproteinof11kDawasdetectedinHepG2andCOS-1cellstransfectedwithpJME.OnlyE(53kDa)proteinwasidentifiedastransfectedwithpJMEusingantiE.HigherlevelofneutralizationantibodiesandtheefficacyofprotectiveimmunitywereinducedwithpJMEimmunization,andweresimilartothoseinducedbyinactivatedJapaneseencephalitisvaccine,butwerebetterthanthoseinducedwithpJE.ItconcludesthattheexpressionlevelfromprMtoEproteinsofJEVisdifferentinvitro,andtheinvitroexpressionefficiencyofpJMEwasbetterthanthatofpiE.FLAG-prMEproteinexpressedbypJMEcouldbecleavedbypeptidasefromhost.TheefficacyofDNAimmunizationiscorrelatedtotheexpressioncharacterizationofrelatedproteinsexpressedinvitro.
简介:AThepurposeofthisinvestigationwastostudythetherapeuticeffectofLamivudineonHBVDNAinperipheralbloodmononuclearcells(PBMC)andserum,andthelevelofcytokinesinserumofthepatientswithchronichepatitisB.Thepatientsweredividedintotwogroups(A=47,B=34),andtreatedbyLamivudine,routinemedicine,respectively.ThelevelsofHBV-DNAinPBMCandserumandcytokineswerealldetectedbeforeandaftertreatment.AfterthetreatmentofLamivndinefor36weeks,thetotalconversionnegativeratesofHBV-DNAinPBMCandserumofthepatientstreatedwithLamivudinewere55.32%(26/47)and61.70%(29/47),respectively.ThetotalnegativeconversionratesofHBV-DNAinPBMCandserumofthepatientstreatedbyroutinemedicinewere26.47%(9/34)and32.35%(11/34),respectively.TherewassignificantdifferencebetweenLamivudinegroupandroutinemedicinegroup(P<0.01).ThenegativeconversionratesofHBeAginserumofthepatientswere46.81%(22/47)and68.09%(32/47)attheendof24weeksand36weeks,andwerehigherthanthoseofroutinemedicinegroup(P<0.05andP<0.01).Thelevelsofalanineaminotransferase(ALT),aspartateaminotransferase(AST),ALT/ASTinserumofthepatientsafterbeingtreatedbyLamivudine,routinemedicineweredown-regulatedto(30.1±9.6)U/ml,(32.3±10.7)U/ml,0.9±0.1and(48.4±10.7)U/ml,(44.7±11.0)U/ml,1.1±0.2.Aftertheanalysisofvariance,thehighsignificantdifferencewasobviousbetweenthetwogroups(P<0.01).ItwasduetothehighlevelsofIL-6,IL-8andTNF-αinchronichepatitisBwhichcouldbedown-regulatedto(250.5±33.3)pg/ml,(153.4±22.2)pg/ml,(232.6±21.2)pg/mlbyLamivudine,whichwasmoreobviousthanthatofroutinemedicine(P<0.01).LamivudinehashightherapeuticeffectonthetreatmentofHBVDNAinPBMCandserumandhasbettertherapeuticeffectthanthatofroutinetherapy.Lamivudinemayalsohavehigherdown-regulatedinflammatoryinfiltrationandsecretioninlocalsitecaused
简介:【摘要】目的: 探讨使用国产和进口试剂对 HCV RNA定量检测的效果。 方法 : 纳入时间为 2018 年 7 月至 2019 年 7 月,选取来我院就诊的慢性丙肝患者 65 例,对所有患者均行使用国产和进口试剂定量检测 HCV RNA水平,现对其 2 种检测方法的结果作分析。 结果: 使用进口试剂定量检测结果相比检测下限而言高出的例数为 58 例,占比为 96.67% ;使用国产试剂定量检测结果相比于检测下限而言高处的例数为 60 例,占比为 92.31% ;经比较组间差异性不显著( p > 0.05 )。 结 论 : 临床在 HCV RNA水平定量检测中使用国产和进口试剂检测结果的一致性和相关性均较高。
简介:TheaimofthisstudyistoinvestigatethefeasibilityandmechanismofhIL-2-preSDNAvaccineaspreventionandtherapeuticapproachagainstHepatitisB.EukaryonexpressionvectorinvolvinghIL-2andpreSgenewasconstructedwithrecombinanttechniqueandtransferredintonormalBALB/cmiceandHBVtransgenicmice(Tg-Mice)respectively.Tnenaseriesofdetectionwereperformed:detectionofanti-preS2,HBsantibodyandHBsAginBALB/cmiceandTg-micewithELISA,quantificationofHBVDNAcopiesinHBVTg-miceserumwithreal-timePCR,determinationofhepatitisdegreewithimmunopathologicalHEstaininganddetectionofliverfunction.Anti-preS1canbedetectedat4^th,6^thand10^thweekininoculatedBALB/cmice.Injectionwithgenegungainedanadvantageovermuscularandsubcutaneousinjectionsinceitacquiredjust1/10inoculationquantity(10μg/mouse).HighestexpressionofIgG2aat4^thweeksuggestedThl-mediatedimmuneresponse,whichfacilitatedHBVcleaning.OfallinoculatedHBVTg-mice,80%ofthemshowedanfi-preS2,HBsantibodypositiveandHBVDNAdecreased,and20%showednegativeforHBsAg.HEstainingtohepatictissueshowedobviousinfiltrationofinflammatorycells,swellingandgranulardegenerationofhepatocytes.Inourstudy,IL-2-preSDNAvaccinewhichcanprovokethehumoralandcellularimmuneresponseandbreaktheimmunetolerancesupportsthedesignationandconstructionofnewvaccineagainstHBVandspecificimmuneremedyforHBVcontinuousinfection.
简介:【摘要】目的:分析荧光定量RT-PCR在流感病毒检测上的应用价值。方法:对23例确诊感染乙型流感病毒的咽拭子、相关机构提供的甲型流感病毒及乙型流感病毒进行检测分析,使用荧光定量RT-PCR检测,分析临床应用的价值及检测的准确率。结果:荧光定量RT-PCR检测乙型流感病毒呈阳性,且结果与其他病毒未见交叉情况。荧光定量RT-PCR检测不同浓度乙型流感病毒敏感度较高,检测的准确率是86.96%(20/23)。结论:荧光定量RT-PCR检测流感病毒的敏感度、准确性较高,可以作为疾病筛查的依据,建议推广应用。
简介:ThepurposeofthisstudywastoconstructaneukaryoticDNAvectorencodingamultipleepitopeantigen(MFC)ofhepatitisCvirus(HCV)andahepatitisBsurfaceantigen(HBsAg),andexploretheeffectofHBsAggeneontheimmunityofHCVmultiple-epitopeDNAconstructinvitroandinvivoinmice.AnHCVDNAvector(pVAX1-HBs-MFC)wasconstructedbyfusingHBsAggenetotheNterminalofanHCVmultiple-epitopeantigengene.ThepVAX1-HBs-MFCwastransfectedintoHEK293TcellsanditsexpressionwasmeasuredbyELISAandWesternblotting.BALB/cmicewereintramuscularlyimmunizedwiththepVAX1-HBs-MFC,andanELISAapproachwasappliedtodeterminethespecificantibodytitersandsubtypesinthemouseserum.Thecross-reactivityoftheantibodieswasalsocheckedwithtwosynthesizedHCVhypervariableregion1(HVR1)peptides.TheIFN-γproductionandcellproliferationofthemousespleencellswereevaluatedbyELISAandMTS(3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium,innersalt)assays,respectively.TheexpressionofpVAX1-HBs-MFCwasdetectableinthetransfectedHEK293Tcells.TheserumantibodyresponsewaseffectivelyelicitedinBALB/cmiceinjectedwithpVAX1-HBs-MFC.ThehighesttiterofantibodyagainstHCV(MFC)was1:1280,andtheratioofIgG2a/IgG1was1.50±0.12atthefifthweekafterfirstimmunization.Moreover,thecollectedmouseserumantibodyhadtheabilitytocross-reactwiththetwosynthesizedHCVHVR1peptides.ThestimulationindexofthemousesplenocytestoMFCwas1.79±0.07,andtheIFN-γlevelwas287±6pg/mlatweek21afterfirstimmunization.ThehighesttiteroftheantibodyincontrolBALB/cmiceimmunizedwithpVAX1-MFCwas1:320,andtheratioofIgG2a/IgG1was1.33±0.11atweek5post-immunization.Furthermore,thestimulationindexofthemousesplenocytescellstoMFCwas1.52+0.06,andtheIFN-γlevelwas225±9.3pg/mlatweek21post-immunization.TheHBsAggenecanenhancetheeffectsofanHCVmultiple-epitope
简介:【摘要】目的: 分析产前定量超声在先天性马蹄内翻足检查中所发挥的价值。 方法: 选择我院在 2017 年 8 月 ~2019 年 3 月产科检查确诊的 先天性马蹄内翻足患儿 43 例设定为研究组,并同期选择产前正常的胎儿 43 例设定为对照组,采用产前定量超声检查测量胎儿的足长、足宽和足长 / 足宽比值指标差异。 结果: 研究组患儿足长、足长 / 足宽数据小与对照组,足宽大于对照组,组间数据对比有意义, P<0.05 。 结论: 先天性马蹄内翻足患儿的足长与足宽与正常胎儿相比具有明显差异,通过产前定量超声检查足长和足宽可诊断 先天性马蹄内翻,具有非常高的超声检查价值。
简介:TherRNAgeneticlocusisfoundinallprokaryoticorganisms,andishighlyconservative,althoughitsrelativelystablevariationsarefoundfrequentlyindifferentbacteria.Theutilityofthislocusasataxonomicandphylogenetictoolhasbeenreportedwidely.Thisstudy,aimedat16SrRNAgene(16SrDNA)andwiththehelpofbiomolecularmethods,attemptedtoachievethegoalofrapididentificationofcommonpathogensInthisstudy,333clinicalisolatedpathogenicbacteriawerecollected。TwopairsofprimerswerechosenandlabeledwithdifferentfluorescentdyesandthenusedtoamplifythegenomicDNAextractedfrombacteria.ThePCRproductswerethendetectedbycapillaryelectrophoresis-singlestrandconformationpolymorphism(CE-SSCP).Inordertopursuehigherresolutionandpeak-separationeffect,ahighefficientseparatingmedium,linerpolyacrylamidedel(LPA),wasputtouseinthisstudy.Finally,everybacteriacolonygenerateddistinctpatternsfromeachother,whichwereeasilytobeusedforidentification.TheseresultsindicatedthatPCR-CE-SSCPwasarapididentificationmethodforbacterialidentification,withtheaspectsofhighefficiencyandhighprecision.Comparedwithtraditionalmethod,thistechnologyisofgreatutilityforclinicaluseespeciallyforitshighsensitivity.