简介:摘要目的观察钛颗粒刺激人血单个核细胞(peripheralbloodmonocytes,PBMC)表达HMGB1、MIF的情况,为临床防治膝关节置换术后无菌松动提供新的思路和方法。方法培养人血单个核细胞,用培养液、钛颗粒及LPS刺激,通过ELISA方法检测HMGB1、NF-?B和MIF表达。结果将分离得到的人血单个核细胞培养后分为3组,分别为空白对照组(PBMC+培养液)、钛颗粒组(0.1%钛颗粒+PBMC+培养液)、阳性对照组(LPS+PBMC+培养液),观察24h后,采用ELISA方法测定HMGB1、NF-?B和MIF表达情况。采用SPSS13.0进行统计学分析,检验水准为?=0.05。结果HMGB1在钛颗粒刺激下表达6.73±0.19ng/ml,空白组表达为1.86±0.24ng/ml,两组对比差异有统计学意义(p=0.000<0.01),阳性组表达为7.51±0.32ng/ml,与钛颗粒组比较差异无统计学意义(p>0.05);MIF在钛颗粒刺激下表达7.74±0.19ng/ml,空白组表达为3.93±0.11ng/ml,两组对比差异有统计学意义(p=0.000<0.01),阳性组表达为9.16±0.55ng/ml,与钛颗粒组比较差异无统计学意义(p>0.05);NF-?B在钛颗粒刺激下表达18.76±1.15,空白组表达为9.57±1.38ng/ml,两组对比差异有统计学意义(p=0.02<0.05),阳性组表达为20.31±1.02ng/ml,与钛颗粒组比较差异无统计学意义(p>0.05);结论钛颗粒刺激人血单个核细胞激活NF-?B通路,且HMGB1、MIF作为免疫炎性因子合成释放增多,参与膝关节置换术后关节无菌松动的发生发展。
简介:摘要目的观察外周血人单个核细胞中MIF、HMGB1、NF-?B的表达,初步探讨关节松动的机制,为临床防治提供新的思路。方法按Ficoll密度梯度分离方法分离静脉血中PBMC,分别用生理盐水(空白对照组)、内固定患者血清(内固定对照组)及关节松动患者血清刺激培养后PBMC,用ELISA和RT-PCR方法检测PBMC中MIF、HMGB1mRNA、NF-?B的表达。应用SPSS13.0软件进行统计学处理数据,P<0.05为差异有统计学意义。MIF的量存在组间明显差异(F=135.633,P=0.000,P<0.05);NF-?B的量同样也存在组间明显差异(F=436.75,P=0.000,P<0.05);HMGB1mRNA在不同组别的表达有明显差异(F=11.935,P=0.006,P<0.05)。MIF与NF-?B表达(r=0.613,P<0.05)具有正相关,NF-?B与HMGB1mRNA表达正相关(r=0.437,P<0.05)。结果MIF、HMGB1、NF-?B在三组表达均存在显著差异,且MIF与NF-?B的表达、HMGB1与NF-?B的表达呈正相关。结论HMGB1通过信号转导通路激活NF-?B的活性,NF-?B可促进单核巨噬细胞合成和分泌MIF,MIF、HMGB1、NF-?B形成细胞炎性因子网络,可能参与关节松动的发生和发展。
简介:目的探讨白细胞介素18(interleukin-18,IL-18)对分泌性中耳炎(otitismediawitheffusion,OME)大鼠中耳微环境中核转录因子nucleartranscriptionfactorskappaB,NF-κB)及T辅助细胞(Thelpercell,Thelpercell)Th2-11hl细胞免疫平衡的影响。方法18只SD大鼠,随机分为OME模型组(A组)、IL-18干预组(B组)和正常对照组(c组)每组各6只(12耳)。A组和B组以卵清蛋白(ovalbumin,ovA)腹腔注射致敏后以OVA耳内激发制成OME模型,c组耳内激发以磷酸盐缓冲液(phosphateBufferedSaline,PBS)替代OVA。IL-18干预组大鼠,于OVA全身致敏及耳内激发的同时,在第1、2、7、8、15、16d给予重组大鼠IL-181μg加生理盐水0.2ml腹腔注射,对照组和OME模型组在相同时间点腹腔注射生理盐水0.2ml替代IL-18。采用HE染色切片观察各组大鼠中耳炎症细胞的变化,免疫组化染色检测中耳黏膜和骨髓腔中IL-4、IFN-γ、NF—KBp65的表达。结果B组中耳Thl型细胞因子IFN-γ含量较A组明显增高(P〈0.05),Th2型细胞因子IL-4含量较A组稍有增高(P,0.05),A组和B组IL一4含量均明显高于c组(P〈0.05);Th2fFhl比值A与B组比较差异无统计学意义(P,0.05),但A组比值明显高于C组(Pc0.05);NF-κBp65蛋白在中耳黏膜和骨髓腔中表达三组间存在显著差异(Pc0.05),B组NF-κBp65阳性细胞比率明显多于A组(Pc0.05),而A组明显多于C组(P〈0.05)。IL-18干预后OME大鼠中耳微环境中编码炎症介质基因表达的转录因子NF-κB活性明显增强,Th细胞过度活化,细胞因子过度分泌,其中IFN-γ合成显著增高,而IL-4合成也出现一定程度的增高,虽然一定程度上纠正了OME大鼠中耳微环境中的Th2/Thl免疫偏移,但是中耳变应性炎症并未得到根本缓解。结论IL-18对Th1和Th2细胞存在双向调节作用参与OME大鼠中耳变应性炎症反应:一方面,刺激Thl细胞�
简介:目的使用RNA-Seq技术分析Miif-m敲除鼠与野生鼠的血管纹转录组,研究Mitf-m基因敲除后的差异表达基因及相关改变的分子机制。方法分别取Miif-m敲除鼠(Mitfmi-△M/mi-△M;MM组)与野生鼠(Miif-m+/+;ww组)的血管纹进行总RNA提取;制备cDNA文库,用IlluminaHiSeq2000测序系统行高通量测序。利用软件Tophat2.2.0将cleanreads比对到参考基因组;分别用软件RSeQS-2.3.2和cufflinks来检测测序结果的均一性和基因表达水平。使用edgeR和DESeq程序筛选差异表达基因(differentialexpressedgenes,DEGs);选择下调DEGs,用KOBAS2.O进行基因本体(geneontology,GO)和京都基因与基因组百科全书(KyotoEncyclopediaofGenesandGenomes,KEGG)代谢途径的富集分析。结果MM组与WW组在参考基因组上mapping的cleanreads数分别为42463888和39718542,分别有88.7%和87.4%的reads是唯一映射,说明RNA-Seq结果可靠。DEGs筛查结果表明,相对于WW组,Mftf-m敲除鼠血管纹有45个DEGs,上调表达基因有7个,下调表达基因有38个。GO富集分析发现下调表达基因与黑色素的合成和代谢过程、色素沉着有关,值得关注的是与内向整流钾离子通道Kcnj10和Kcnj13相关。KEGG富集分析显示下调表达基因主要富集于黑色素生成通路。结论RNA-Seq分析拓展了Mftf-m基因调控网,为探究Mitf-m突变所致听力-色素综合征的分子机制及特异性研究Mitf不同转录子的功能奠定了基础。
简介:NuclearfactorkappaB(NF-κB)isoneofthebest-characterizedtranscriptionfactorsplayingimportantrolesinmanycellularresponsestoalargevarietyofstimuli,includinginflammatorycytokines,phorbolesters,growthfactors,andbacterialandviralproducts.TheaimofthisstudyistodemonstrateNF-κBexpressioninthemousecochleaanditsenhancementinresponsetolipopolysaccharides(LPS)andkanamycin(KA)treatment.MethodsKAtreatmentconsistedofsubcutaneousKAinjectionsat700mg/kgtwiceadaywithaneight-hourintervalbetweenthetwoinjectionsfor3or7days.ForanimalsintheLPStreatmentgroup,asingledoseof0.3mgLPSdissolvedin0.2mlsterilesalinewereinjectedintobothbullaethroughthetympanicmembraneandkepttherefor3hours.Animalsinthecontrolgroupreceivedsubcutaneoussalineinjectionfor7days.Followingimmmunohistochemichalprocessingwithrabbitpolyclonalanti-NF-κBp65antibodies,cryosectionsofthecochleawereexaminedforexpressionofNF-κBp65invariousstructuresinthecochlea.ResultsNF-κBp65expression,identifiedbypresenceofbrownreactionproductscharacteristicofDABimmunohistochemistry,wasvisibleinthespiralligament,spiralprominence,tectorialmembrane(TM),spiralganglionandnervefibers.RelativelyweakNF-κBp65expressionwasalsovisualizedintheorganofCorti.WithintheorganofCorti,theinnerhaircells(IHC),outerhaircells(OHC),innerpillarcells(IP),outerpillarcells(OP),Deiter'scells(DC),andBoettcher'scellsexhibitedstrongerstainingthantheinnersulcuscells,Hensen'scells(HC)andClaudius'cells.NoNF-κBp65expressionwasseeninthenucleusoftheIHCandOHC.NF-κBp65expressionwasincreasedinanimalsexposedtoLPSorKA,demonstratingsignificantdifferencesinthestainingbetweencontrolanimalsandLPS/KA-treatedanimals.NF-κBp65expressionwasnotsignificantlydifferentbetweenLPStreatedandKAtreatedanimalsorbetween3and7daysinKA-treatedanimals.Conclusio
简介:ObjectiveChronictinnitusisahighlyprevalentconditionandhasbeenhypothesizedtoresultfromaninnatedisturbanceincentralnervousserotonergictransmission.Giventhefrequentcomorbiditywithmajordepressionandanxiety,wearguethatcandidategenesforthesedisordersarelikelytooverlap.Thepresentstudyaddressesthegeneencodingforthe5-HT1Areceptorasaputativeriskfactorfortinnitus.MethodsIn88subjectswithadiagnosisofchronicsubjectivetinnituswhounderwentadetailedneurootologicalexamination,theentire5-HT1AgenewasamplifiedusingoverlappingPCRproducts.Ampliconswerecustomsequencedbidirectionallyandwerescreenedforvariantsinmultiplealignmentsagainstthehumangenomereference.ResultsWeidentifiedasynonymousC>Texchangeatresidue184(Pro)in7/88subjects,butdetectednomissensevariantsinthepopulationunderstudy.Specifically,thefollowingresidueswerefullyconserved:16(Pro),22(Gly),28(Ile),98(Val),220(Arg),267(Val),273(Gly),and418(Asn).DiscussionThepresentdatacountagainstthecausationofchronictinnitusbyachangeinthe5-HT1Areceptor'saminoacidsequence.However,theallelefrequencyforthe184Prominorallele(0.04)reachedtwicethefrequencyreportedincontrolcohortsfromthesameethnicity.Additionalinvestigationsareinvitedtoclarifytheroleofthe5-HT1Apolymorphisminlargersamples,andtocontrolforcomorbidaffectivedisorders.
简介:目的评价通过B超测量面肌厚度的方法诊断面肌萎缩的可行性,并测量青年人面肌厚度的正常值,分析面肌厚度正常值的相关因素。方法选取34名健康青年志愿者,记录所有志愿者的年龄、性别、身高、体重及头围5种相关因素,用B超按设定方法测量每位志愿者双侧的额肌、降口角肌和降口唇肌,对测量结果及5种相关因素进行统计学分析。并从34名志愿者中随机选取5名,在1周后和2周后分别重复测量3组面肌,比较3次测量结果有无差异。结果①34名志愿者经B超测量3组面肌厚度,双侧额肌、降嗣角肌和降口唇肌厚度无显著差异(P〉0.05)。面肌厚度平均值为:额肌左侧(1.60±0.21)mm,右侧(1.62±0.22)mm;降口角肌左侧(2.60±0.50)mm,右侧(2.62±0.51)mm;降口唇肌左侧(1.51±0.23)mm,右侧(1.52±0.23)mm。②随机选取的五5名志愿者重复测量3次,3次面肌厚度测量结果相近,无统计学差异(P〉0.05)。③按性别分组,女性18例,男性16例,组间比较:男性3组面肌厚度大于女性,差异有统计学意(P〈0.05)。④体重指数与所测量的3块面肌厚度呈正相关性(P〈0.05),额肌、降日角肌、降口屠肌与体重指数的相关系数分别为0.391、0.459及0.447;头围与面肌厚度无明显相关性。⑤选定的年龄段内(20-30岁)3组面肌厚度与年龄无相关性(P〉0.05)。证明20-30岁年龄段的青年人,面肌厚度不受年龄影响,该年龄段人群面肌厚度具有相对稳定性。结论①B超测量面肌厚度具有可重复性和很好的稳定性;②正常青年人(20-30岁)双侧额肌、降口角肌和降口唇肌厚度无明显差异:③不同性别的正常青年人(20-30岁)面肌厚度有明显差异,男性的额肌、降口角肌、降口唇肌均较女性厚;④正常青年人(20-30岁)面肌厚度跟体重指数呈正�
简介:1病例报道患者,男,44岁。因右面部肿块1个月于医院就诊,行CT检查发现右侧上颌窦肿块,于2015年9月4日入院。右侧面部颧骨下方可触及约4cm×4cm×3cm的略韧肿块,无压痛。鼻腔检查未见异常,颈部无异常。CT显示右侧上颔窦前壁大小约3cm×2cm×2cm肿块,内有钙化斑块(图1)。
简介:Thedevelopmentandplasticityofcentralauditorysystemcanbeinfluencedbythechangeofperipheralneuronalactivity.However,themolecularmechanismparticipatingintheprocessremainselusive.Brain-derivedneurotrophicfactor(BDNF)bindingwithitsfunctionalreceptortropomyosinreceptorkinaseB(TrkB)hasmultipleeffectsonneurons.Hereweusedaratmodelofauditorydeprivationbybilateralcochlearablation,toinvestigatethechangesinexpressionofBDNFandTrkBintheauditorycortexafterauditorydeprivationthatoccurredduringthecriticalperiodforthedevelopmentofcentralauditorysystem.Reversetranscription-quantitativepolymerasechainreaction(RTqPCR)andimmunohistochemistrymethodswereadoptedtodetectthemRNAandproteinexpressionlevelsofBDNFandTrkBintheauditorycortexat2,4,6and8weeksaftersurgery,respectively.ThechangeintheexpressionofBDNFandTrkBmRNAsandproteinsfollowedsimilartrend.Inthebilateralcochlearablationgroups,theBDNF-TrkBexpressionlevelinitiallydecreasedat2weeksbutincreasedat4weeksfollowedbythereductionat6and8weeksaftercochlearremoval,ascomparedtotheage-matchedshamcontrolgroups.Inconclusion,theBDNF-TrkBsignalingisinvolvedintheplasticityofauditorycortexinanactivity-dependentmanner.
简介:摘要目的通过观察超顺磁性壳聚糖质粒(pDsVEGF165Red1-N1)明胶微球(SPCPGM)与磷酸钙骨水泥的复合支架在外加振荡磁场下修复颅骨缺损的作用,探讨其成骨效果。方法将50只新西兰大白兔随机分为5组,并制成颅内颅骨缺损模型,分别植入超顺磁性壳聚糖质粒明胶微球(SPCPGM-VEGF165)/多孔磷酸钙骨水泥(CPC)复合支架、超顺磁性壳聚糖明胶微球((SPCPG)/多孔磷酸钙骨水泥(CPC)复合支架、多孔磷酸妈骨水泥(CPC)支架,经振荡磁场处理。于术后2周、4周、8周、12周时通过大体、X线检测、组织切片观察血管化及成骨情况,图像分析系统分析、求积仪测量对骨缺损处成骨情况进行测定,评估各组成骨的情况。结果在振荡磁场下超顺磁性壳聚糖质粒明胶微球(SPCPGM-VEGF165)/多孔磷酸钙骨水泥(CPC)复合支架组支架开始降解吸收速度、血管化、成骨作用优于对照组。结论在振荡磁场下超顺磁性壳聚糖质粒明胶控释微球局部控释增加了持续作用时间,同时促进了质粒VEGF165细胞转染,从而促进新生骨血管化及成骨的作用,可用于颅骨缺损修复。
简介:目的研究10号染色体缺失张力蛋白磷酸酶(phosphataseandtensinhomologuedeletedonchromosometen,PTFEN)、磷酸化Akt(P—Akt)及核转录因子-KB(NF—KB)在中耳胆脂瘤上皮中的表达,探讨P13K(phos—phatidylinositol-3-kinase,磷脂酰肌醇-3激酶)-Akt信号通路在中耳胆脂瘤上皮细胞过度增殖机制中的可能作用。方法采用免疫组织化学SP法(辣根酶标记链霉卵白素连接法,streptavidin—peroxidaseconjugatedmethod)检测30例中耳胆脂瘤组织标本与15例正常外耳道皮肤标本中PTEN、P—Akt及NF—KB蛋白的表达。结果PTEN蛋白阳性表达主要定位于上皮细胞核,其在中耳胆脂瘤上皮中阳性表达率为36.7%,明显低于正常外耳道皮肤组的9313%(P〈0.01);P—Akt蛋白阳性表达主要定位于上皮细胞胞质,其在中耳胆脂瘤上皮中阳性表达率为70.0%.明显高于正常外耳道皮肤组的26.7%(P〈0.01);NF—KB蛋白阳性表达定位于上皮细胞核.其在中耳胆脂瘤上皮中阳性表达率为63-3%,明显高于正常外耳道皮肤组的20.0%(P〈0.01)。在30例中耳胆脂瘤上皮组织中,PTEN分别与P—Akt、NF—KB蛋白的表达之间呈显著负相关(P〈0.01),而P—Akt和NF—KB蛋白的表达呈显著正相关(P〈0.01)。结论PTEN、P-Akt和NF—KB在中耳胆脂瘤上皮的异常表达可能在胆脂瘤的发生、发展过程中起重要作用。胆脂瘤上皮中P13K—Akt信号通路的激活可能参与了胆脂瘤上皮细胞过度增殖机制。
简介:ObjectivesToinvestigatetheexpressionofhistamineH1receptors(H1R)inthevestibularnucleusofbrainsteminratsandtheroleofH1Rinmotionsickness(MS).MethodsAtotalof24healthySprague-Dawleyratsweredividedrandomlyintofourgroups(n=6each)whichdeterminediftheanimalswouldreceiveinductionofMSordrug(promethazine)treatment:MS(-)/Drug(-);MS(+)/Drug(-);MS(-)/Drug(+at0.25mg);andMS(+)/Drug(+).MSwasinducedbycomplexmotionstimulationandtheconditionedtasteaversionwasusedasabehavioralindicatorofMS.Thevolumeof0.15%sodiumsaccharinsolution(SS)intakewithin45minutesaftermotionstimulationwasmeasured.H1Rinthevestibularnucleuswasexaminedbyimmunofluorescencestaining.TheexpressionofH1Rproteininbrainstemtissueatvestibularnucleuslevelwasdetectedbywesternblot.ResultsThemeanSSintakevolumeintheMS(+)/Drug(-)group(8.8ml)wassignificantlylessthanthatoftheMS(-)/Drug(-)group(15.1ml)(P<0.01).ThemeanSSintakevolumeoftheMS(-)/Drug(+)group(14.8ml)wassimilartothatoftheMS(-)/Drug(-)group.ThemeanSSintakevolume(9.6ml)oftheMS(+)/Drug(+)groupwasmorethanthatoftheMS(+)/Drug(-)group(P<0.01),butlessthanthatoftheMS(-)/Drug(-)grouporMS(-)/Drug(+)group(P<0.01).ImmunofluorescencestainingshowedpositiveexpressionofH1Rinthevestibularnucleusofbrainstemandtheexpressionwasenhancedbymotionstimulation.WesternblotanalysisshowedthatH1Rproteinexpressedinthebrainstemtissueatvestibularnucleuslevelandtheexpressionalsoincreasedsignificantlyaftermotionstimulation.TheMS-inducedincreaseofH1Rwasnotaffectedsignificantlybypromethazine.ConclusionsH1RsexistinthevestibularnucleusinratsandH1Rexpressionisup-regulatedbymotionstimulation,butnotaffectedbypromethazine.ThefindingsindicatethatthehistaminergicsystemisinvolvedinMS.Promethazine,asanH1Rblocker,mayplayitsanti-MSrolebycompetingthebindingsiteon