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: 研究临床血脂生化检验中应用分级检验方法的临床检验效果。 方法: 选取我院检验科收集的 300 份血脂生化样本作为研究对象,两等分后分为对照组与观察组,每组各 150 份血液样本,对照组采用拉网式检验,观察组采用分级检验方法,比较两组 TG 、 TC 、 HDL 、 LDL 、载脂蛋白 A 、载脂蛋白 B 的阳性检出情况。 结果: 两组在 TG 、 TC 、 HDL 阳性检出率方面比较, P>0.05 ,无明显统计学差异;观察组的 LDL 、载脂蛋白 A 与载脂蛋白 B 阳性检出率高于对照组,数据比较 P<0.05 ,存在明显统计学意义。 结论: 分级检验方法可使血脂生化检验的阳性检出率大幅提高,为临床高血脂等疾病诊断提供可靠性依据,具有推广价值。
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:探究和讨论分级检验法应用于肾脏生化检验的临床效果,旨在为鉴别和诊断疾病提供科学依据。方法:选取我院检验科于 2018年 10月 ~2019年 10月间收治的 100例行肾脏生化检验的患者为观察对象,将其按照随机抽签法分为对照组与观察组,两组患者人数均为 50例。对照组患者采取常规检验法接受检验,观察组患者则采取分级检验法接受检验。回顾性分析 2组患者的临床资料,比较检验效果。结果:观察组患者检验血肌酐水平( Scr)、胱抑素水平( CysC)及血尿素氮水平( BUN)的阳性率分别为 60.00%、 44.00%及 60.00%,均显著高于对照组的 6.00%、 8.00%及 4.00%,经统计学分析,组间差异具有统计学意义( P<0.05)。结论:与常规检验法相比,分级检验法在肾脏生化检验中具有较高的准确率,临床价值高,推荐应用。