简介:Objective:ToexploretheeffectsofnuclearM-CSFontheprocessoftumorigenesis.Methods:FunctionalpartofM-CSFcDNAwasinsertedintoaneukaryoticexpressionplasmidpCMV/myc/nuc,whichcanaddthreeNLStotheC-terminaloftheexpressedproteinanddirecttheproteinintothecellnuclei.TheconstructedplasmidwastransferredintoNIH3T3cellsandthecellcloneswereselectedbyG-418selection.CellclonesstableexpressingtargetproteinwereidentifiedbyRT-PCR,ABCimmunohistochemistryassayandWesternblot.Cellgrowthkineticsanalysesthroughgrowthcurves,celldoublingtime,MTTtestandanti-senseoligodeoxynucleotide(ASODN)inhibitingcellgrowthtestwereperformedtoidentifycellsproliferationpotential.Results:Thetransfectedcellsshowedelevatedproliferationpotentialoverthecontrolcells.Conclusion:AbnormalappearanceofM-CSFinnucleuscouldenhancecellproliferation,whichsuggeststhatcytokineisoformswithincellnucleusmightplaytranscriptionfactor-likerole.
简介:摘要目的比较IgA肾病患者不同病理分级的血清IgA、C3及IgA/C3水平;探讨血清IgA、C3及IgA/C3水平在IgA肾病病理分级评估中的意义。方法选择我院320例经肾穿刺活检病理检查确诊为原发性IgA肾病患者,用散射比浊法检测患者血清IgA、C3水平及按Lee氏分级标准评估IgA肾病病理分级。结果Lee分级为Ⅰ、Ⅱ级与Lee分级为Ⅲ、Ⅳ、Ⅴ级患者IgA水平分别为(2.75±1.10)vs(2.85±1.26)g/L,(P﹥0.05);血清C3水平分别为(1.18±0.33)vs(1.04±0.29)g/L,(P﹤0.05);血清IgA/C3率分别为(2.71±1.19)vs(3.24±1.69)g/L,(P﹤0.05)。结论IgA/C3比值在评估IgA肾病病理分级有重要参考意义。
简介:TheeffectofTPA,apotenttumorpromoter,onSSV-NIH3T3cellsinserum-freemediumwasinvestigated.TPAstimulatedDNAsynthesisofSSV-NIH3T3cellsonthethirddayofcultureinSFM.InSDS-PAGFofmediumconditionedbyTPA-treatedSSV-NIH3T3cells(inSFM+TPA),theamountsoffourproteinsof31.0Kd,28.5Kd,25.5Kdand13.5Kdstrikinglyincreasedoverthatofnon-TPA-treatedcounterpart(inSFM).ThePDGF-likeactivitywasalsodetectedinCMofSFM+TPA.WheninsulinandEGFweredrownofftheSFM+TPA(SFM-Ins-EGF+TPA),TPAlostitsabilitytostimulateDNAsynthesisofSSV-NIH3T3cellsonthethirddayandSDS-PAGEoftheconditionedmediumshowedthattheamountsofthefourproteinsnotedabovegratelyreduced.However,cellsinSFM-Ins-EGF+TPAwereinalmostthesamegrowthconditionascellsincompleteSFM+TPAonthethirddayofculture.Resultswerediscussedinthepaper.
简介:FiveC3/C3fluoroquinolonedimerstetheredwithafusedheterocyclicringofs-triazolo[2,1-b][1,3,4]thiadiazolederivedfromantibacterialquinolonesweresynthesizedandcharacterized,andtheirinvitroantitumoractivityagainstL1210,CHOcelllineswasevaluatedviatherespectiveIC50values.
简介:摘要目的旨在探讨C3a-C3a受体(C3aR)在常染色体显性多囊肾病(ADPKD)进展中的作用及机制。方法收集ADPKD患者切除肾组织和PKD1敲除小鼠多囊肾组织,观察C3a、C3aR及F4/80在肾组织中的表达;利用脂多糖(LPS)和白细胞介素4(IL-4)分别刺激巨噬细胞,检测各组细胞C3aR、TNF-α、分型标志物和相关信号通路的表达及机制;使用C3aR抑制剂SB290157(1 mg/kg)处理PKD1敲除小鼠,观察肾脏病理、囊肿相关指标和肾功能变化。结果C3a及C3aR在ADPKD患者和PKD1敲除小鼠肾组织中表达显著上调(均P<0.05),C3aR与F4/80共定位于小鼠多囊肾组织中的巨噬细胞上。体外培养M1型巨噬细胞C3aR表达显著上调(P<0.05),C3a刺激后M1型巨噬细胞表达iNOS、TNF-α和IL-6 mRNA显著上调(均P<0.05),分泌TNF-α增多,说明C3a不仅影响M1型巨噬细胞自身炎性因子表达,还影响其周围炎症微环境;此外,C3a激活M1型巨噬细胞Akt信号通路显著上调(P<0.05)。与模型对照组比较,SB290157治疗组小鼠血Scr、BUN水平、囊肿指数、双肾重/体重(2KW/BW)均显著降低(均P<0.05),小鼠肾组织中C3a、C3aR水平及p-ERK、p-P65通路蛋白表达显著下调(均P<0.05)。结论多囊肾组织中增多的C3a通过C3aR引起巨噬细胞浸润和活化,进而促进ADPKD进展,其机制可能通过Akt活化、TNF-α产生增多所致。C3aR拮抗剂是治疗ADPKD的一个潜在研究方向。