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简介：ToinvestigatetheeffectsofoverallalkaliofatraditionalChinesemedicine“Tongbiling”(brucineandstrychninealkaloidsinmain)onthecytokinesexpressioninTh1andTh2cellsinthesynovialfluidofpatientswithrheumatismarthritisandtheirsignalpathway,themononuclearcellsinthesynovialfluid(SFMC)ofpatientswereisolatedbyFicoll-Hypaquegradientcentrifugation,andtheCD3^+CD69^+andCD3^+HLA-DRantigenwereanalyzedbyflowcytometryincomparisonwiththoseoftheperipheralblood.TherestofcellswereculturedafterresuspensionwithRPMI1640culturemedium.Phorbol12,13-dibutyrate(PDB)andionomycinwereaddedsuccessivelyintotheculturewithvariousconcentrationofoverallalkaliTongbiling(TBL).After4hofcultivation,theexpressionofIFN-γandIL-4inCD3^+cellswereanalyzedbyflowcytometry.TheinfluenceofoverallalkaliTBL(100mg/I,)ontheintracellularcalciumwasinvestigatedafterFluo-3/AMlabelingandstimulationwithPDBandionomycinat1,2,4and10min,andtheinfluenceofTBLontheexpressionofCD3^+CD69^+cellsweredeterminedwithstimulationofPDBfor24hinthewholebloodlymphocytesculture.ItwasfoundthatthepercentageofTcellsbearingCD69wassignificantlyup-regulated(77%),whilethatofTcellsbearingHLA-DRwas44%inthesynovialmononucleatedcells.AfterPDBandionomycinstimulation,theexpressionofIFN-7inCD3~cellswereup-regulated,buttherewasnochangeontheexpressionofIL-4inCD3^+cells,indicatingthatratioofTh1/Th2wassignificantlyincreasedandThcellsdifferentiatetoThlcellsinmainly.FourconcentrationsofoverallalkaloidofTBI,(200mg/L,100mg/L,50mg/L,25mg/L)coulddown-regulatedtheexpressionofIFN-γinCD3^+cellsandtheTh1/Th2ratioobviously,butalltheconcentrationsoftheoverallalkaloidshadnoeffectontheexpressionofIL-4inCD3^+cells.100mg/Lconcentrationoftheoverallalkaloiddidnotdown-regulatetheintracellularcalciumlevel.Eachconc

简介：Toinvestigatethemutationsintheupstreamregulatoryregion(URR)ofhumanpapillomavirustype16(HPV-16)fromthecervicalcancerbiopsiesinXinjiangUygurwomenanditsrelationshiptothehighincidenceofcervicalcancerinthesouthernXinjiang,thetissueDNAwasextractedfromthecervicalcancerbiopsies,andtheURRsegmentofHPV-16DNAwasamplified,sequencedandanalyzed.Thereafter,thepolymorphismofURRinHPV-16wasthenanalyzed.ItwasdemonstratedthatthepositiveratedetectedforthepresenceofURRinHPV-16was89.47%(17/19).ComparedwiththepreviouslypublishedsequenceinURRofprototypeHPV-16,somemutationsweredetectedinthesequenceofURR.Themutationsin17URRfragmentsofHPV-16couldbedividedinto11patterns(XJU-1toXJU-11)atnucleicacidlevel,inwhicheachofXJU-1andXJU-4accountedfor23.53%(4/17),andotherpatternsofmutationaccountedfor5.88%(1/17).IncomparisonwiththeURRofprototypeHPV-16,theDNAidentityofthesepatternswas98.50%-99.68%.Inthese17URRfragments,twopointmutationsoccurredatposition7192(GtoT)andposition7520(GtoA)andtheyappearedtobeconstantinXinjiangarea.ThesetwomutationswereubiquitousintheAsia-Americantypeandconferredstronginfectionactivityandcarcinogenicityofthisvirus.Inaddition,themutationsatposition7729(AtoC),position7843(AtoG)andposition7792(CtoT)couldenhanceitstranscriptionactivityconsiderably.ItisconcludedthatsomemutationsoccurinURRgeneofHPV-16inthecervicalcancerbiopsiestakenfromUygurwomeninXinjiangarea,suggestingthatcertainrelationshipexistsamongthemutationsinURRofHPV-16,thephylogenyofHPV-16andthehighincidenceofcervicalcancerinsouthernpartofXinjiangarea.

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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.