简介:目的:探讨鬼臼毒素纳米脂质载体(POD-NLC)诱导人永生化阴道上皮细胞(VK2/E6E7)凋亡的作用及机制.方法:取对数期生长VK2/E6E7细胞,分别加入POD-NLC和Caspase抑制剂Z-VAD-FMK共同培养24小时.实验分为4组:A组:空白对照组;B组:0.25μg/mLPOD-NLC+Caspase抑制剂组;C组:1μg/mLPOD-NLC+Caspase抑制剂组;D组:空白NLC+Caspase抑制剂组.采用流式细胞仪检测ROS水平,qPCR检测AIFmRNA表达,蛋白印迹检测各组细胞AIF、cyt-C蛋白表达.结果:B组、C组均可以上调ROS、AIF、cyt-C水平,与A组及D组相比均有统计学差异(P值均〈0.05);B组与C组AIFmRNA及AIF蛋白表达无统计学差异(P值均〉0.05);ROS、cyt-C各组间比较均有统计学意义(P值均〈0.05).结论:鬼臼毒素纳米脂质载体可以通过非Caspase依赖途径诱导永生化阴道上皮细胞凋亡.
简介:Objective:Tostudytheexpressionofactivatedepi-dermalgrowthfactorreceptor(EGFR)andtranscrip-tionfactorE2F(E2F)inCondylomaAccuminata(CA)patients.Methods:ImmunofluorescenttechniqueswereusedtoinvestigatetheexpressionofactivatedEGFRandE2FinCApatients.Results:TheexpressionofactivatedEGFRonthemembraneofepithelialcellsinCAlesionswassig-nificantlygreatercomparedtoexpressionleversinthecontrolgroup(P<0.01).Moreover,theco-expres-sionofactivatedEGFRandE2Fwassignificantlyin-creasedcomparedtothecontrolgroup(P<0.01).Conclusion:Ourobservationssuggestthatthein-creaseinactivatedEGFRexpressionmaystimulatehyperplasiainCApatientsthroughtheactivationoftranscriptionfactorE2F.
简介:高免疫球蛋白E综合征(HIES)是一种罕见的免疫缺陷性疾病,表现为血清IgE水平升高、皮肤湿疹、反复皮肤感染和特征性的肌肉骨骼表现。作者报道2例在本院就诊的患者并回顾当前文献。患者1是一18个月大的非裔美国男孩,伴有复发性葡萄球菌冷脓肿、肺炎和菌血症。该患者皮肤严重湿疹化,最终并发疱疹性湿疹。在选用敏感抗生素环孢素A(5mg/kg)短期治疗全身性感染后,皮炎改善,随后改为长期口服甲氧苄啶一磺胺甲恶唑。患者2是一15岁高加索男孩,长期患有HIES。该患者从3岁开始即维持每周3次注射1干扰素和每月静脉注射免疫球蛋白,并预防性应用抗生素和小剂量氟康唑。
简介:目的研制人乳头瘤病毒16型(HPV16)L1/E7CTL重组杆状病毒载体。方法以包含HPV16标准病毒株的质粒pHPV16为模板,采用PCR技术扩增HPV16主要衣壳蛋白L1(1-471aa)的基因序列,同时人工合成E7蛋白CTL表位E749-57的编码序列,将两个目的基因逐步连接构建重组质粒pUC19HPV16L1/E7CTL,HPV16L1/E7CTL片段经质粒pFastBac1转位至BacmidDNA后通过PCR鉴定重组杆粒。结果成功构建了包含融合基因HPV16L1/E7CTL片段的克隆质粒puc19HPV16L1/E7CTL,制备了L1/E7CTL重组杆状病毒载体杆粒BACMIDDNA。结论采用分子克隆技术构建了包含L1/E7CTL的重组杆粒BACMIDDNA,有利于下一步融合蛋白的表达,制备一种能够更有效激发CTL杀伤作用的嵌合病毒样颗粒疫苗,为疫苗研制打下基础。
简介:目的检测活化型表皮生长因子受体(EGFR)和转录调节因子E2F在尖锐湿疣中的表达。方法采用免疫荧光方法观察活化型EGFR和E2F在尖锐湿疣(CA)皮损中的表达情况。结果活化型EGFR在CA皮损角质形成细胞膜上的表达较正常对照组显著增强(P<0.01)。而且,活化型EGFR和E2F在CA皮损中的双重阳性表达较正常对照组显著增强(P<0.01)。结论活化型EGFR表达的增强导致核内转录因子的激活而参与CA的增殖病变。
简介:Objective:ToobtainrecombinantTreponemapallidumsubsp,pallidum(TP17KD)lipoproteininlargequantitiesbyamplificationandtofurtherpurifyantigensforlaboratorydiagnosisofsyphilisanddevelopmentofasyphilisvaccine.Method:TheTppl7lipoproteingenewasamplifiedfromtheTP(strainNichols),andthenitwasrecombinatedintoaplasmidpMAL-2candclonedwithinE.coli12-TB1.ThehostbacteriacontainingrecombinantplasmidswereinducedwithIPTG.TheTpp17KDlipoproteingenewasamplifiedbyus-ingPCRandpositivecloneswerescreenedwithdoubledigestionandPCR.RecombinantplasmidsweretransformedintoE.coliandtheE.colicarryingrecombinantplasmidswereinduced.TheexpressionofTP17KDwasdetectedbysodiumdedecylsulfate-polyacrylamidegelelectrophoresis(SDS-PAGE)andimmunoblot.Results:GelstainingwithCoomassieblueG-250showedthattheinducedE.colicarryingrecombinantplasmidcouldproduce60KDfusionproteinathighlevels.Gelscanningshowedthat17KDproteinexpressioninE.coliaccountedfor10%oftotalcellularprotein.Therecombinantproteinantigenreactedwiththeseraofsyphilispatients.Conclusion:Ourstudylaysacornerstonefordevelopingnewtechniquesoflaboratorydiagnosisforsyphilisandnewvaccines.Preliminaryclinicalapplicationshowedthatthefusionproteincouldbeusedforthediagnosisofsyphilis.
简介:寻常狼疮是皮肤结核中最常见的类型,约占所有皮肤结核患者的半数。本病易侵犯儿童和青少年,好发于面部,影响容貌。我们用异烟肼、乙胺丁醇和利福平联合治愈寻常狼疮2例,报告如下。
简介:Inordertoacknowledgethemulti-infectiondataofSTDpatientsandimprovetheprophylaxisandtherapyforSTDs,297patientsand30healthypeoplewereexaminedusingtheHSV-2plasmaantibody.TheresultsshowedthatallkindsofSTDpatientswereinfectedbyHSV-2indifierentratios.thehighestratiooccurringinsyphilispatients.
简介:Objective:TounderstandthechangeofTh1/Th2andTc1/Tc2balanceinpatientswithsecondaryearlysyphilis.Methods:PeripheralbloodThandTccellsubpopulationswereexaminedbythree-colorflowcytometryin30patientswithsecondaryearlysyphilisand20healthysubjects.Results:PercentagesofCD3^+,CD4^+TcellsandtheratioofCD4^+/CD8^+andNKcellsinsyphiliticpatientsweresignificantlylowerthaninthecontrolgroup(P<0.001),butnostatisticaldifferencewasfoundinpercentagesofCD8^+andCD19^+cellsbetweenthetwogroups(P>0.05).ThepercentageofTh1cellsandratiosofThl/Th2andTcl/Tc2insyphiliticpatientswereclearlydecreasedascomparedtothoseincontrois(P<0.001-0.01),whereaspercentagesofTc0andTc2cellsinsyphiliticswashigherthanthoseincontrols(P<0.001).Conclusion:Therecouldbeadisturbanceofcellularimmunityandtherelativelyintactfunctionofhumoralimmunityinsecondarysyphilis.ItispostulatedthatthepolarizationtowarddevelopmentofTh2andTc2cellsandtheirassociatedcytokinesmightbeinvolvedintheincompleteclearanceoftreponemesandthedevelopmentofchronicinfection.
简介:Objective:Toconstructarecombinantplasmidcontainingtheoutermembraneprotein2(Omp2)geneofChlamydiatrachomatisandexpressOmp2inE.coli.Methods:Theomp2geneofC.trachomatisserovarDwasclonedintopQE30vectorfollowingPCRamplificationfromgenomicDNA.E.coliM15transformantswereinducedtoexpressthefusionproteinbyIPTGandtheproductwasidentifiedbySDS-PAGEandWesternblot.Results:ConfirmedbyenzymecleavageanalysisandDNAsequencing,acorrectrecombinantplasmidpQE30/omp2wasconstructed.Thefusionproteinfromthetransformantswasapproximately60kDainsizeinSDS-PAGEanalysis,whichcouldspeciallyreactwithanti-6×HismousemonoclonalIgGantibodies.Conclusion:WesuccessfullyexpressedOmp2inE.coilM15,providinganefficientandsimplesystemforassayingtheimmunologicalpropertiesofOmp2.
简介:Objective:Tostudythecellularimmunitystatusofpatientswithrecurrentgenitalherpes.Methods:Serumlevelsofinterlukin-2anditssolublereceptorandinterlukin-6weremeasuredbyELISAin34patientswithrecurrentgenitalherpes.Results:SerumlevelsofIL-2andIL-6weresignificantlydecreasedinpatientscomparedtohealthycontrols(P<0.01),andthelevelofsIL-2Rwassignificantlyincreasedinpatientswithrecurrentgenitalherpes(P<0.01).Therewerenosignificantdifferencesinallvariablesamongstpatientsregardingrelapsestageandremissionstage(P>0.05).Conclusion:Therewasacellularimmunedeficiencyinpatientswithrecurrentgenitalherpes.
简介:ObjectivesTocomparetheefficiencyoffourcommercialELISAkitsindetectingtype-specificHSV-2IgGantibodies.MaterialsandMethods:Atotalof125subjects,including105withgenitalulcers,and20controlswithoutanyhistoryofSTDswererecruitedfromtheSTDclinicfordetectionoftype-specificHSV-2IgGantibodywithdifferentkits.FourkindsofcommerciallyavailableELISAkits,includingQuidaHSV2IgGELISA(AifulangBiochemCo.Ltd.,Hangzhou),TORCH-HSV2IgG(JingmeiBiotechCo.Ltd.,Shanghai),Captia^TMHSV2IgG(Trinitybiotech,USA)andHerpeSelectT^TM2ELISAlgG(Focustechnologies,USA)wereusedforevaluation.WesternBlotassaywasperformedasagoldstandard.Results:ComparedtoWesternBlotresults,thesensitivityandspecificityofthekits(QuidaHSV2,TORCHHSV2,Captia^TMHSV2andHerpeSelect^TM2)were13.1%and98.4%,7.5%and100%,100%and11.1%,87.7%and96.7%,respectively.Thepositivepredictivevalue(PV)andnegativePVofthefourkitswere88.9%and54.3%,100%and55.5%,55.6%and100%,96.2%and89.2%,respectively.TheareasundertheROCcurveofthreekits(QuidaHSV2,Captia^TMHSV2andHerpeSelect^TM2)were0.885(0.822-0.948),0.825(0.747-0.902),0.974(0.950--0.998),respectivelyConclusion:TheperformanceofHerpeSelect^TM2isthebestamongthefourkits.Theresultsalsoindicatethatthecommerciallyavailablekitsfordetectionoftype-specificHSV2antibodyshouldbere-evaluatedintermsoftheirvalidity.
简介:Objective:Toinvestigatetherolesofinterleukin-2(IL-2)andinterleukin-10(IL-10)inpathogenesisofearlysyphilis.Methods:TheserumlevelsofIL-2andIL-10in48patientswithearlysyphilisweredetectedbyABC-ELISA.Results:(1)ThelevelofIL-2inthepatientswithearlysyphiliswassignificantlyhigherthanthatinhealthycontrols,whilethatofIL-10waslower(P<0.001andP<0.001).(2)ThelevelsofIL-2andIL-10werealmostidenticalinpatientswithprimaryandsecondarysyphilis(P>0.05),aswellasbetweendif-ferentRPRtiters(P>0.05).(3)Aftertherapy,thelevelofIL-2decreasedmarkedly(P<0.05),whilethatofIL-10increase(p>0.05).(4)AsignificantcorrelationwasfoundbetweentheserumlevelsofIL-2andIL-10(r=0.5385P<0.05).Conclusions:Th1up-regulationoccursinpatientswithearlysyphilis,andplaysanactiveroleinfightingagainstTPinfection.