简介:Thepurposeofthestudyistoestablishafluorescencequantitativereversetranscriptionpoly-merasechainresponse(FQ-RT-PCR)methodforthequantitativedeterminationofIL-2mRNAandIL-4mRNAinThcells,withwhichtheThcellsstatusofthepatientswithgynaecologicaltumorsandchronicrenalfailure(CRF)canbeanalyzed.IL-2cDNAandIL-4cDNAwereprepared,andtheplasmidpMD18carryingIL-2cDNAorIL-4cDNAfragmentwasconstructedandclonedasthetemplateforquantitativedetermination.Theprimersandprobeslabelledwith6-carboxy-fluorescein(FAM)and6-carboxy-tetrarnethylrhodamine(TAMRA)wereprepared,andtheexperimentalconditionswereoptimizedtosetuptheFQ-RT-PCRmethodforquantitativedeterminationofIL-2mRNAandEL-4mRNA.Thcellsenrichedfromperipheralbloodmononuclearcells(PBMCs)of20healthyvolunteers(HVs),16gynaecologicalbenign(GB)cases,18gynaecologicalmalignant(GM)tumorcasesand16chronicrenalfailure(CRF)patientsweretestedforIL-2mRNAandIL-4mRNAbyFQ-RT-PCR.Thehouse-keepinggeneβ-actinwasusedastheinternalcontrolgeneoftheexperiment.Thestandardcurveforlogconcentrationofseriesofquantitativetemplatesvsthresholdcycle(CT)wasestablishedbylinearregression,andthelinearrangewas102-107copies/μl.TheimprecisiontestshowedtheCVofinter-assayandintra-assayofahighcontentsamplebyFQ-RT-PCRwere7.8%and12.5%,respectively.TheCVofinter-assayandintra-assayofalowcontentsamplewere10.8%and19.5%,respectively.TheIL-2mRNAexpressionsinThofthepatientswithgynaecologicalmalignanttumor(comparedwiththeHVsandthepatientswithgynaecologicalbenigndisease)andinThoftheCRFpatients(comparedwiththeHVs)weredeclinedsignificantlyandatthesametimetheIL-4mRNAexpressionincreasedsignificantly(P<0.001).Asimple,sensitiveandaccurateFQ-RT-PCRmethodforthequantitativedetectionofIL-2mRNAandIL-4mRNAhasbeenestablished.TheIL-2mRNAandIL-4m
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 : 探讨白介素 - 6( IL- 6 ) , 白介素 - 8( IL- 8 ) 与胎膜早破 的关系 。 方法 : 收集 2018年 1月 -2019年 1月的 160例胎膜早破孕妇作为病例组 ; 随机抽取同期正常孕妇160例作为对照组 。 两组孕妇入院后即抽取静脉血提 取血清 ,采用 ELISA 法检测血清中IL- 6 、IL- 8的含量 ,统计分析病例组和对照组感染绒毛膜羊膜炎的几率 ,采用 western blot 测定胎盘组织中 IL- 6 、IL- 8含量 。结果 : 病例组血清中 IL- 6 ,IL- 8 的含量明显高于对照组 , 差异有统计学意义 ;病例组感染绒毛膜羊膜炎的几率明显高于对照组 , 差异有统计学意义 ; 结论 :IL- 6 ,IL- 8 可能全程参与妊娠过程 ,因此联合监测 IL- 6 、IL- 8 的含量对于预测胎膜早破新生儿的预后有一定的作用 , 尤其在降低新生儿死亡率 、预防远期并发症等方面有一定价值 。
简介:目的:分析过敏性鼻炎(allergic rhinitis,AR)患者外周血CD4 +Th17细胞中IL-18、IL-18结合蛋白a(IL-18-binding protein a, IL-18BPa)和IL-18受体α(IL-18 receptor α,IL-18Rα)的表达水平及过敏原对其表达的影响。 方法:选择2019年10月—2020年9月锦州医科大学附属第一医院过敏反应科门诊AR患者45例和该医院体检中心体检者23例(健康对照组)为研究对象。根据皮肤点刺试验结果,将患者分为阳性组(AR组,24例)和阴性组(nAR组,21例)。收集受试者血液样本,流式细胞术检测过敏原对CD4 +Th17细胞中IL-18、IL-18BPa和IL-18Rα蛋白质表达水平的影响。Bioplex检测血浆中IL-17A水平,并分析其与IL-18 +Th17细胞比例的相关性。 结果:与健康对照组比较,AR组患者CD4 +Th17细胞和IL-18 +Th17细胞比例均增加( P<0.01),IL-18BPa +Th17细胞比例降低( P
简介:Triptolideisanatural,biologicallyactivecomponentderivedfromChineseherbTripterygiumWilfordiiHookF.(TWHF)whichiseffectiveintheclinicaltreatmentofautoimmunediseases,however,themechanismsbywhichtriptolideexertsimmunosuppressionremainfullyunderstood.Theprimaryofthisstudyistodemonstratewhethertriptolidecanaffectphenotype,cytokineproductionandallogeneicTcell-stimulatorycapacityofdendriticcells(DCs)whicharecriticalintheinductionofimmuneresponseortolerance.PhenotypicanalysisshowthattriptolidedoesnotaffecttheexpressionofMHC(Ia^b),CD80,CD86andCD40ofDCstimulatedwithornotLPS,butsignificantlyinhibitsIL12p70productionbyDCinadose-dependentmanner.Triptolide-treatedDCsexhibitareducedcapacitytostimulateproliferationofallogeneicCD4^+Tlymphocytes.Therefore,triptolide-mediatedimmunosuppressionmaydue,inpart,totheinhibitionofIL-12p70productionandimpairmentofallogeneicTcell-stimulatorycapacityofDCs.Ourresultsmayprovideapossiblemechanisticexplanationfortheeffectivenessoftriptolideinthetreatmentofautoimmunediseases.
简介:TheexpressionofIL-4inaratmodelofchronicpulmonaryinfectionbiofilmformationinducedbyPseudomonasaeruginosawasinvestigated,inwhichSPFWisterratswereinfectedviatracheawith0.1mlP.aeruginosastrainPAO579(10~9CFU/ml)inalginatebeadsortheplanktonicformofthisbacterialstrain(10~9CFU/ml),andon3,7and14dafterinfection,thebacteriologicalandpathologicalchangeswereobservedaswellastheexpressionofthecytokineIL-4wasdetermined.ItwasdemonstratedthatthecountofCFUperlungtissueincaseofbacteriainalginatebeadswassignificantlyhigherthanthatofbacteriainplanktonicform,withmoreseveregrosspathologicchangesandinflammatoryreactionsinthealginatebeadgroupincomparisonwiththatoftheplanktonicforms(P=0.002,P=0.004andP=0.002,respectively).Inaddition,theexpressionofIL-4inthealginatebeadgroupwasalsohigherthanthatintheplanktonicform(P=0.02,P=0.02andP=0.022,respectively).Apositivecorre-lationbetweenthelevelofIL-4expressionandthegrosslungpathologyinalginatebeadgroupexistedasdemonstratedbysimpleregressionanalysis(r=0.78,P<0.02).Itisconcludedthatthechronicpul-monaryinfectionwithbiofilmformationinducedbyP.aeruginosatendstohavetheprioritytotheTh2immuneresponse.
简介:摘要:国际糖尿病联盟(IDF)提出糖尿病的治疗目标不能仅局限于控制血糖水平,还要将提高病人的生活质量作为重要指标。1DF提出的五项基本措施,即饮食治疗、体育锻炼、药物治疗、糖尿病教育和自我血糖监测,在21世纪仍然需要对糖尿病进行综合治疗。此外,还应注意同时积极处理心脑血管疾病危险因子如高血压、脂代谢紊乱等。
简介:【摘要】目的:分析对 老年 2型糖尿病 (T2DM)患者采用利拉鲁肽治疗的 临床效果。方法:选取我院 2018年 2月 -2019年 4月收治的 110例 老年 2型糖尿病 (T2DM)患者为分析对象,给予所有患者利拉鲁肽进行治疗,经治疗 8 周后,对患者治疗前与治疗后的空腹血糖、餐后 2h 血糖、体重指数以及胰岛 β 细胞 功能指数进行对比。 结果:经所有患者 8 周治疗后所有相关临床指标均优于治疗前,治疗前后指标对比差异存在显著临床可比性 ( P<0.05)。 结论:临床上对 老年 2型糖尿病 (T2DM)患者采用利拉鲁肽治疗效果显著,在改善患者胰岛功能以及血糖时具有较高的安全性,值得在临床推广应用 。
简介:Analysisofthefrequencyofantigen-specificcytotoxicTlymphocytes(CTLs)exvivoislargelydependentontheuseofMHC/peptidetetramers.However,thelatterreagentshavenotbeenwidelyavailable,mostlikelybecauseoftheircostlyandtime-consumingproduction.InthisreportweutilizedaneconomicstrategytoconstructHLA/peptidetetramerswithrecombinantpeptide-linkedβ2microglobulin(β2m).TheHLA-A2-restricted,melanomaantigenMARTl-derivedpeptideMARTI27-35(AAGIGILTV)wasfusedtotheNterminusofhumanβ2mthrougha15-aminoacid(aa)-longlinkerbeforebeingrefoldedwiththerecombinantbiotinylatedHLA-A2heavychainectodomain.Theresulted2-component(2C)monomerwasthentetramerizedwithphycoerythin-labeledstreptavidin.Theexperimentalresultshowedthatthe2CHLA-A2/MARTI27-35monomerwasshowntobindtotheHLAclassⅠcomplex-specificmonoclonalantibodyW6/32andtheHLA-A2/MARTI27-35complex-specificsinglechainantibodyfragment(scFv)8.3,suggestingthecorrectnessofitsspecificity.Furthermore,the2CHLA-A2/MARTI27-35tetramerdetectedaspecificCD8^+TcellpopulationinHLA-A2-restrictedmelanomainfiltratinglymphocytesastheconventional3CHLA-A2/MARTI27-35tetramer.Theyieldof2CHLA-A2/MARTI27-35monomerwas2.5timesmorethanthatoftheconventional3Cmonomer.Takentogether,thesedataindicatethattheHLA-A2/MARTI27-35tetramercanbegeneratedconvenientlythroughtheuseofMARTI27-35peptide-β2mfusionproteins,whichcanfacilitatethemonitoringofHLA-A2-restricted,MARTl-specificCTLresponsesinpatientswithmelanoma.
简介:【摘要】目的:分析综合护理干预在 2型糖尿病肾病护理中所取得的护理价值。方法:选择我院在 2017年 4月 ~2019年 5月诊治的 2型糖尿病肾病患者 84例作为护理对象,将患者均分为观察组 42例和对照组 42例,观察组应用综合护理干预,对照组应用常规护理,对比两组患者护理后血糖和肾功能指标、患者护理前后情绪状态。结果:观察组患者的血糖和肾功能指标均优于对照组, P<0.05;在护理前,两组患者负面情绪评分相似,经过护理后,观察组患者负面情绪评分降低,数据对比有意义。结论:在 2型糖尿病肾病患者的护理中应用综合护理干预可显著改善病情,缓解不良情绪,取得了积极的护理价值,值得推广。
简介:ToexploretheantiviraleffectandmechanismofpolysaccharidefromSpirulinaplatensis(PSP)onherpessimplexvimstype2(HSV-2),astandardstrainofHSV-2(333strain)wasusedtoinvestigatetheantiviraleffectofPSPinvitro.PSPinvariousconcentrationswasappliedtodifferentstagesofHSV-2replicationcycle.Finally,thevirusinfectivity(TCID50),cytopathiceffect(CPE),andMTTstainingmethodforviablecells(MTTassay)wereusedasmarkerstoevaluatetheeffectofPSPonHSV-2.ThequantityofHSV-DNAwasdetectedbyreal-timefluorescencequantitativePCR(FQ-PCR).TheHSV-2infectedVerocellultrastructureswereobservedbytransmissionelectronmicroscopy(TEM).TheresultsshowedthatPSPhadlittlecytotoxiceffectonVerocells,itcouldnotdirectlyinactivateHSV-2infectivity.PSPnotonlyinterferedinadsorptionofHSV-2toVerocellsbutalsoinllibitedHSV-2biosynthesisinthecells.FQ-PCRresultsshowedthattheinhibitoryrateonHSV-DNAalsoincreasedinadose-dependentandtime-dependentmanner.TEMalsoconfirmedthatPSPexhibitedpronouncedinhibitoryeffectonHSV-2.Inconclusion,theantiviraleffectofPSPonHSV-2maybeattributedtotheinhibitionofvimsadsorption,vimsreplicationandsynthesisincells.