简介:摘要:目的:慢性髓系白血病经酪氨酸激酶抑制剂治疗后疗效较前明显改善,但仍有复发或急变可能,且病情凶险。故判断病情可能预后极其重要。本研究探讨吲哚胺2,3-双加氧酶在慢性髓系白血病各期中的表达及意义。方法:于我院收集病例,分为正常对照组、CML-CP组, CML-BP/AP组。用ELISA方法检测IDO水平。结果:对照组IDO水平显著低于CML-CP组, CML-BP/AP组IDO水平明显高于CML-CP组,且差异均具有统计学意义。结论:慢性髓系白血病患者中,IDO水平高表达,表明IDO水平与慢性髓系白血病发生发展密切相关。
简介:InordertoinvestigatewhetherlipoxinA4(LXA4)hasanantagonisticeffectonlipopolysaccharide(LPS)-inducedsynthesisofinterleukin(IL)-1β,IL-6andIL-8inratpulmonarymicrovascularendothelialcells(PMVEC),andtoexplorethemolecularmechanismsofsignalpathwayinLXA4actions,culturedPMVECweretreatedwithLPS,withorwithoutpreincubationwithLXA4.ProteinsofIL-1β,IL-6andIL-8insupernatantwereanalyzedbyenzyme-linkedimmunosorbentassay(ELISA).ExpressionsofmRNAofIL-1β,IL-6andIL-8weredeterminedbyRT-PCR.Expressionsofphosphorylationofphosphoinositide3-kinase(PI3-K)andmyeloiddifferentiationfactor88(MyD88)wereanalyzedbyWesternblot.ActivitiesofDNA-bindingofnuclearfactor-kappaB(NF-κB)andactivatorprotein-1(AP-1)weremeasuredbyelectrophoreticmobilityshiftassay(EMSA).TheresultsshowedthatLPSinducedproductionofIL-1β,IL-6andIL-8inratPMVECviaMyD88/PI3-K/NF-κBandAP-1pathway-dependentsignaltransduction.LPS-stimulatedexpressionofPI3-K,activitiesofNFκBandAP-1,secretionofproteinandexpressionofmRNAofIL-1β,IL-6andIL-8butnotMyD88expressioninPMVECwereinhibitedbyLXA4inadose-dependentmanner.Inconclusion,LXA4inhibitssynthesisofIL-1β,IL-6andIL-8bydown-regulationofPI3-K/NF-κBandAP-1signalpathwayinPMVEC.
简介:ToclarifytheroleofAPOBEC3G(A3G)incellulardefenseagainsthepatitisBvirus(HBV),theexpressionofA3GinnormalhumanliverandtheregulationoftheA3Gexpressioninhep-atomacellline(HuH-7)wereinvestigated.ExpressionlevelofAPOBEC3smRNAinhumanliverwasdeterminedbyRT-PCR.HuH-7andHepG2cellsweretreatedwithvariousconcentrationsofIFN-α(0U/ml,100U/ml,500U/ml,1000U/ml)for12h.ThemRNAlevelsweremeasuredbyaquantitativeRT-PCR,theresultswerenormalizedrelativetothespecimenswithoutIFN-αstimulation.TotalproteinofHuH-7cellstreatedwithvariousconcentrationsofIFN-αfor48hwassubjectedtoWesternblotanalysis.Forreportergeneassay,HuH-7cellsweretransfectedwiththereporterplasmidscontainingIRF-Esitesanditsmutantswithdifferentlengths.Thenthecellsweretreatedwithorwithout1200U/mlIFN-aforadditional12h(1000U/ml)after24hoftransfection,andthecelllysatewaspreparedandassayedforluciferaseactivity.ItwasfoundthatnormalhumanliverexpressedthemRNAofA3G.A3GmRNAexpressioninHuH-7andHepG2cellswereup-regulatedbyIFN-αstimulationinadose-depen-dentmanner.WesternblotanalysisindicatedthatA3GproteinexpressionwasalsoenhancedbyIFN-αstimulation.SequenceanalysisshowedtheexistenceofputativesitesofIFNregulatoryfactorelement(IRF-E)in5'regionofA3Ggeneupstreamtheinitiationcodon.IFN-αstimulationresultsin6-to8-foldincreaseinluciferaseactivityincellstransfectedwiththeplasmidcontainingIRF-Esitesofthe5'upstreamsequences,whereasluciferaseactivitydidnotchangeincellstransfectedwiththeplasmidcontainingmutantIRF-EsitesorwithoutIRF-Esites.Asaconclusion,A3Gareexpressedinnormalhumanliver.A3Gexpressionwasup-regulatedbyIFN-αstimulationinhepatomacellsandcouldbeinvolvedinhostdefensemechanismsagainstHBV.ERF-Esitein5'regionofAP0BEC3Ggeneupstreamtheinitiationcodonplaysanimportantroleinthisp