简介：目的观察Nucleus24CA型人工耳蜗植入后电极阻抗、行为反应阈值（T-level,T级）及最大舒适级（C-level,C级）的变化规律,分析其内在联系,探讨其对术后调机的指导意义。方法对81例植入Nucleus24CA型人工耳蜗患儿,分别在术中、术后1、2、6个月进行电极阻抗阈值测试,收集术后对应T、C值,并对其变化规律及相关性进行统计学分析。结果电极阻抗值术中检测最低,术后1月开机最高,此后逐渐减低（P〈0.01）;自蜗顶至蜗底各通道间电极阻抗值无显著差异（P〉0.05）。各电极通道T值、C值随术后时间延长逐渐增高（P〈0.05）,并与电极阻抗值呈线性相关。结论测定电极阻抗值是评估人工耳蜗刺激电极状态的有效手段;术后2月应同时调试T值及C值,此后则应对C值进行重点调试。

简介：目的用细胞学方法,分析线粒体DNA12SrRNA基因中C1494T突变在氨基糖甙类抗生素聋发病机理中的作用.方法从携有线粒体DNAC1494T突变的母系遗传性氨基糖甙类抗生素性耳聋的中国大家系选择部分成员,另外从遗传背景相同的正常中国人群选择对照个体,分别建立淋巴细胞系;并通过细胞融合技术,将淋巴细胞系的线粒体分别融合到缺乏线粒体DNA的p0206细胞中,建立相应的转线粒体细胞系;家系成员与对照个体的淋巴细胞系和转线粒体细胞系,分别在不含/含有氨基糖甙类抗生素(巴龙霉素)的培养液中培养,以倍增时间(doublingtime,DT)作为细胞生长特性的评价标准,通过计算在正常和含有氨基糖甙类抗生素的培养液中倍增时间的比值,比较氨基糖甙类抗生素对细胞生长的影响.结果携有线粒体DNAC1494T突变家系成员较对照个体的淋巴细胞系的倍增时间比值平均增加了24%,但不同家系成员的细胞倍增时间比值的增加程度不同,自10%至50%不等;而当细胞核遗传背景相同后,家系成员较对照个体的转线粒体细胞系的倍增时间比值增长30%,并且来自不同表型的家系成员的细胞倍增时间比值基本相同.结论线粒体DNAC1494T突变可以造成细胞对氨基糖甙类抗生素的超敏性,但其效应要受到核基因的调控.

简介：Objective:Toinvestigateimmune-relatedgeneticbackgroundinbilateralsuddensensorineuralhearingloss(SSNHL).Casereportandmethods:Thecaseisa45-year-oldmanpresentingwitha7-yearhistoryofbilateralprofoundSSNHL.Bloodbiochemicaltestingdemonstratedincreasedlevelsoftotalcholesterol(5.88mmol/L).TestsforhepatitisBshowedapositiveantibodyagainstthehepatitisBcoreantigen.ComplementC3wasbelowthenormalvalue,andcomplementC4andIgGwereinthelowerrangeofnormalvalues.CTimagesshowedanormalinnerearandvestibularaqueductbutroundwindowmembranousossificationonbothsides.Atotalnumberof232immuneassociatedgenesweresequencedusingthenextgenerationsequencingtechnique.Results:Mutationsweredetectedin5genes,includingthephosphoinositide3-kinasecatalyticsubunitdelta(PIK3CD),caspaserecruitmentdomain-containingprotein9(CARD9),complementfactorH-related(CFHR2),immunoglobulinlambda-likepolypeptide1Protein(IGLL1),andtransmembranechannel-likegenefamily8(TMC8).InthePIK3CDgene,aC896Tsubstituteinexon7wasdetected.Thismutationcausesprimaryimmunodeficiencyandisanautosomaldominantdisease.Conclusion:ThePIK3CDC896TmutationresponsibleforprimaryimmunodeficiencymaycontributetotheonsetofbilateralSSNHLwithsubsequentrapidprogression.

简介：Aratmodelofchronictympanicmembraneperforationwasdevelopedtobeusedinthesearchofnewmaterialsforthesealingoftheseperforations.AlongitudinalstudywascarriedoutinratssubjectedtoincisionalmyringotomyfollowedbytheapplicationofmitomycinCaloneorwithdexamethasone.Ratswerecheckedatdays3,7,10,14andweeklythereafteruntilperforationclosure,forupto6months.Theadditionofdexamethasoneisakeycomponentinordertoobtainachronicopening.Myringotomiestreatedwithsalinehadameanhealingtimeof8.5days.At8weeks,between62.5%and77.7%oftympanicmembranestreatedwithmitomycinCanddexamethasoneremainedperforatedandat6monthsthisnumberfellto21.4%.Thistechniqueisabletomaintainmosttympanicmembraneperforationspatentforatleast8weeks.Thisratmodelisadequateforitsuseinpreclinicalortranslationalresearch.

简介：ObjectiveToinvestigateTcellactivationfollowingfacialnerveaxotomizationandlatentneuroimmunologicmechanismsintraumaticfacialparalysis.MethodsAmurinemodeloffacialnervetransactionwasused.LymphocytesfromcervicalandmesentericlymphnodesinBABL/cmiceatspecifictimeswerecollectedandexpressionratesofCD69onTcellswereassessedbyflowcytometry.ResultsInfiltratingTcellsweredetectedaroundthefacialneuronsinthefacialnervenucleusinmicewhosefacialnervewastransected.ImmunofluorescentstainingshowedrecruitmentofactivatedTcells.Threedayspost-facialnervetransection,theexpressionrateofCD69onTcellsfromcervicaldraininglymphoidnodes(CDLNs)wassignificantlydifferentfromthatonTcellsfrommesentericlymphnodes(MLNs)(P=0.0457),whereasthelatterwassimilartothatinanimalsundergoingshamsurgeriesandthatinblankcontrolanimals(p=0.2817and0.2724,respectively).Twoweekspost-nervetransection,theTcellCD69expressionratefromCDLNsremainedatahigherlevelandthanthatinthesham-operationanimals(p=0.0007).Attwoweeks,CD69expressionrateonTcellsfromMLNswasalsoup-regulatedanddifferentcomparedwiththesham-operationanimalsandwithitselfatthreedayspost-operation(p=0.0082and0.0133,respectively).ConclusionTcellsappeartobeactivatedandup-regulatedinCDLNsfollowingfacialnervetransection.ThereisevenevidenceofTcellactivationinMLNsat2weekspost-nervetransection.Thissuggestesanalterationofimmuneresponsefromlocaltogeneralimmunityintheacutestageoffacialnervetrauma,whichmayhelpcoordinatingandcontrollingthescalesandorientationoftheneuroimmuneresponseduringthepathogenesisandprogressionoffacialnervetrauma.

简介：Objective:EvaluatingtheauditoryfunctioninpatientswithchronichepatitisCtreatedwithsofosbuvirandribavirin.Methods:Thisstudyinvolved80patientswithchronichepatitisCwhoagreedtoreceivesofosbuvirandribavirin.Allparticipantsweresubjectedtobaselineotologicalandaudiologicalassessmentjustbeforetreatment.Theaudiologicalassessmentincludedstandardpuretoneaudiometry,extendedhighfrequencyaudiometry,immitancemetryandotoacousticemissions(OAEs)(transientanddistortionproduct).Accordingtobaselinehearingthresholdmeasurements,thestudypopulationwasdividedinto2groups.Group1included42patientswithnormalhearingsensitivity(250e8000Hz),andGroup2included38patientswithsensorineuralhearingloss.After24weeksoftherapy,otologicalandaudiologicalassessmentswererepeatedandcomparedbetweenthetwogroupsandbeforeandaftertherapy.Results:Post-treatmenthearingthresholdevaluationshowednosignificantdifferencefrompretreatmentevaluationatalltestedfrequencies.Therewasnostatisticallysignificantdifferencebetweenpreandpost-treatmentotoacousticemissionsresults.Conclusion:TherapywithsofosbuvirandribavirininchronichepatitisChasnonoticeableeffectsoncochlearfunctions.

简介：摘要目的通过对28例行预防性T形肠造口术病人的观察及术后回访，总结了该新治疗方法的护理方法。方法回顾性分析2009年4月至2014年4月我科28例急性肠梗阻及超低位直肠癌行预防性T形肠造口术患者的临床资料。围手术期给予健康宣教、造口护理及出院指导。结果28例患者均康复出院，平均住院天数11.2天。1例回肠造口行临时性缝合闭合后粪便自造口漏出，经凡士林纱布覆盖压迫后痊愈。1例出现轻度造口周围皮炎，经造口治疗师及时对症处理后缓解。结论对急性肠梗阻及超低位直肠癌行预防性T形肠造口术的患者，术前做好心理护理及术前准备，术后密切观察病情变化，预防并发症发生，能使患者平稳度过围手术期，降低术后并发症的发生，可提高手术治疗效果。

简介：目的应用流式细胞技术探讨面神经损伤急性期T细胞功能状态及其病理生理意义，为揭示外伤性面瘫的神经免疫机制提供理论依据。方法制备面神经轴切损伤模型小鼠，分别于损伤后3天、2周分离小鼠肠系膜淋巴结（mesentericlymphnode，MLN）、术侧颈部引流淋巴结（cervicaldraininglymphnode，CDLN）细胞，进行双色免疫荧光标记。以流式细胞术分析T细胞上CD69分子的表达情况。结果手术后3天，面神经损伤组颈部引流淋巴结T细胞CD69表达的百分率与相应手术对照组相比较差异有显著性（P=0．0457）；而神经损伤组、手术对照组肠系膜淋巴结T细胞CD69表达的百分率与正常小鼠相比较差异无显著性（P值分别为0．2817、0．2724）。面神经轴切损伤后2周，神经损伤组CDLN的T细胞CD69表达的百分率仍然维持在较高水平，神经损伤组MLN的T细胞CD69表达的百分率出现上调，与相应对照组及术后3天时相比较差异均有显著性（P值分别为0．0082、0．0133）。结论面神经轴切损伤3天、2周时，颈部引流淋巴结存在T细胞活化并上调；在2周时。肠系膜淋巴结也出现低水平的T细胞活化。提示面神经损伤急性期。伴随着一个局部免疫应答向全身免疫应答转化的过程，在一定程度上有利于机体协调控制免疫应答的规模与方向。

简介：颈胸部各种创伤、气管切开或气管插管后造成的气管狭窄是后天性气管狭窄主要原因。近年来随着应用机械呼吸疗法,这种原因所引起的气管狭窄也日渐增多。气管狭窄为不可逆转的、进行性加重的病变,唯一有效的治疗方法是外科手术切除狭窄组织,恢复气管管腔通气。因为病程是进行性加重,也有骤然发生气管完全梗阻的危险，所以手术宜尽早进行。

简介：ObjectiveToinvestigatetheearlychangeofcochlearribbonsynapsesoninnerhaircellsinresponsetoaminoglycosideototoxicity.MethodsC57BL/6Jmicereceivedintraperitonealinjectionofgentamicin(100mg/kg/day),andtheapicalcoilorganofCortiwasexaminedonthe4th,7thand10thday(n=10).Litter-mateswithoutgentamicintreatmentservedascontrols(n=10).RIBEYEonthepresynapticmembraneandAMPAreceptorsonthepostsynapticmembranewerelabeledwithCtBP2orGluR2/3respectively.Threedi-mensionreconstructionwasconductedusingthe3DSMAX8.0software.ResultsTherewerenodisruptionsofouterorinnerhaircellsinallgroups.However,thenumberofribbonsynapsesoncochlearinnerhaircellsincreasedsignificantlywithin7daysaftergentamicinexposure(P<0.01),followedbyasignificantde-creaseafter7days.ConclusionDuringtheearlystageofaminoglycosideototoxicity,increasedpopulationofcochlearribbonsynapsesmayindicateasignificantdown-regulationofsynapticfunction.

简介：ObjectiveTostudycharacteristicsofhearinglossafterexposuretomoderatenoiseexposureinC57BL/6Jmice.MethodsMaleC57BL/6Jmicewithnormalhearingatageof5-6weekswerechosenforthisstudy.Themicewererandomlyselectedtobestudiedimmediatelyafterexposure(GroupP0),or1day(GroupP1),3days(GroupP3),7days(GroupP7)or14days(P14)afterexposure.Theirbeforeexposureconditionservedasthenormalcontrol.Allmicewereexposedtoabroad-bandwhitenoiseat100dBSPLfor2hours,ABRthresholdswereusedtoestimatehearingstatusateachtimepoint.ResultsABRthresholdelevationwasseenateverytestedfrequencyatP0(P<0.01).Elevationathigh-frequencies(16kHzand32kHz)wasgreaterthanatlowerfrequencies(4kHzand8kHz,P<0.05).FromP1toP14,ABRthresholdscontinuouslyimproved,andtherewasnosignificantdifferencebetweenP14andbeforeexposure(P>0.05).ConclusionThereisafrequencyspecificresponseto100dBSPLbroad-bandwhitenoiseinC57BL/6Jmice,withthehigh-frequencybeingmoresusceptible.HearinglossinducedbymoderatenoiseexposureappearsreversibleinC57BL/6Jmice.

简介：Objective:Todeterminewhetheranew-bornchildfromafamilycarryingadeafnessgeneneedscochlearimplantationtoavoiddysphoniabyscreeningandsequencingadeafness-relatedgene.Results:BothscreeningandsequencingresultsconfirmedthatthenewbornchildhadanormalGJB2genedespitethefactthatshehasabrothersufferingfromhearinglosstriggeredbyanallelicGJB2c.176del16mutation.WeclonedtheGJB2genesderivedfromtheirrespectivebloodgenomicDNAintoGFPfusedplasmidsandtransfectedthoseplasmidsintothe293Tcelllinetotestforgenefunction.WhilethemutatedGJB2gene(GJB2c.176del16)ofherdeafbrotherwasfoundtobeunabletoformthegapjunctionstructurebetweentwoadjacentcells,thebabygirl’sGJB2generanintonosuchproblems.Conclusion:ThescreeningandsequencingaswellastheGJB2genefunctiontestsinvariablyshowedresultsconsistentwiththeABRtestedhearingphenotype,whichmeansthatthechild,withanormalwildtypeGJB2gene,doesnotneedearlyinterventiontopreventherfromdevelopinghearinglossanddysphoniaatalaterstageinlife.

简介：目的构建含有小鼠Smad4基因和增强型绿色荧光蛋白基因（EGFP）的慢病毒病毒表达载体,为今后应用该重组慢病毒介导的内耳基因导入和相关的聋病基因治疗奠定实验基础.方法利用基因重组、限制性内切酶酶切及基因测序的方法,构建并鉴定pLenti6.3-Smad4-IRES2-EGFP真核表达质粒;利用脂质体介导转染的方法,将pLenti6.3-Smad4-IRES2-EGFP转染导入293T细胞,荧光显微镜下观察EGFP基因表达情况,利用实时荧光PCR的方法检测小鼠Smad4基因mRNA水平的表达情况.结果经PCR鉴定和基因测序证实了小鼠Smad4基因序列与基因bank中的序列相一致.pLenti6.3-Smad4-IRES2-EGFP质粒转入293T细胞后,荧光显微镜下可见有绿色荧光蛋白表达.293T细胞经慢病毒感染后,其小鼠Smad4基因mRNA表达量增加了66427倍.病毒滴度经测定为2.5×108TU/ml.结论成功地构建了含有小鼠Smad4基因的慢病毒表达载体,并能在293T细胞中表达.

简介：目的探讨老年性耳蜗毛细胞损害与中药复方健耳剂两种喂药方法干预的作用。方法选择1月龄C57BL/6J小鼠22只用于本实验,其中4只小鼠每日饮用自来水直到出生后3个月作为幼龄对照组;6只小鼠每日饮用自来水直到出生后7个月作为老年性聋对照组;6只小鼠每日自动饮用中药复方健耳剂直到出生后7个月;另6只小鼠每日自动饮用同样中药至4个月后改用每日人工灌服直到出生后7个月。各组动物实验到期终止后,取耳蜗进行全耳蜗基底膜铺片,将全耳蜗内、外毛细胞计数结果输入计算机并应用耳蜗图软件进行耳蜗毛细胞密度对比分析,其中选择基底膜上重要的病变区间的毛细胞密度进行统计学分析。结果3月龄对照组小鼠耳蜗外、内毛细胞缺损仅仅出现在耳蜗底回钩端区域;7月龄对照组外、内毛细胞缺损从底回基底膜起始端扩展到距离耳蜗顶端约40%区域;7月龄中药灌服组和自动饮用组动物的内、外毛细胞缺损范围和程度相似,均显著比7月龄对照组为轻（P〈0.001）。结论中药复方健耳剂能够有效延缓C57BL/6J小鼠老年性耳蜗毛细胞损害的发生和发展,两种喂药方式所起作用相同（P〉0.05）,其药理机制可能与其改善微循环,清除活性氧,保护线粒体等作用相关。

简介：目的探讨不同周龄C57BL/6小鼠内耳形态学及其ABR阈值变化。方法取C57BL/6小鼠3周、4周、12周、26周各10只,听性脑干反应(ABR)测试双侧2、4、8、16、20kHzABR阈值。采用基底膜铺片MyosinⅥ、Neurofilament免疫组化染色,观察耳蜗毛细胞和神经丝的变化。扫描电镜观察耳蜗毛细胞及其静纤毛随年龄的变化。结果随着年龄增长,C57BL/6小鼠各频率ABR阈值明显提高,顶转和底转内毛细胞缺失逐渐增多,神经丝染色渐淡,毛细胞静纤毛逐渐发生数量减少、增粗融合、倒伏等变化。到26周龄时已达到重度聋,各频率较3周组有显著统计学差异。顶转和底转内毛细胞有连续缺失,外毛细胞完全缺失,内毛细胞只有残存的少量静纤毛,粗细不均,倒伏明显。结论本研究对国产C57BL/6小鼠的内耳形态进行观察,明确了其ABR阈值和内耳毛细胞的变化规律,为用国产C57BL/6小鼠进行老年性聋研究提供了依据。

简介：目的探讨幼年和10月龄C57BL／6J小鼠听力及初级听皮质（AI）中凋亡抑制蛋白XIAP的年龄相关变化。方法检测两组C57BL/6J小鼠听性脑干反应（ABR）；免疫组织化学法染色检测两组C57BL/6J初级听皮质神经元凋亡抑制蛋白XIAP的表达情况。结果与幼年C57BL／6J小鼠相比，10月龄C57BL/6J的ABR反应阈值更高，凋亡抑制蛋白XIAP的表达显著减少。结论随年龄增长C57BL/6J小鼠的听力减退，同时C57BL／6J小鼠大脑初级听皮质凋亡抑制蛋白表达减少，XIAP的表达水平可能与C57BL/6J小鼠听力减退有关。

简介：目的比较腺病毒、脂质体、纳米材料三种不同类型的载体携带目的基因对293T细胞的转染效率及细胞毒性的大小,筛选理想的基因载体。方法用重组腺病毒、Lipofectamine^TM2000、Entranster^TM—D纳米材料为转染载体,携带含有增强型绿色荧光蛋白（EGFP）报告基因的Math1-EGFP基因及真核表达质粒pRK5-Math1—EGFP，按照转染试剂盒说明优化其转染条件，分别转染293T细胞。24小时后在共聚焦显微镜下观察转染结果并计数阳性细胞率，采用RT—PCR法检测转染细胞中Math1基因mRNA的表达，用MTT法检测不同转染条件下，不同转染载体对293T细胞的细胞毒性。结果经优化转染条件，重组腺病毒载体转导的细胞中荧光蛋白表达几乎为95％以上,Lipofectamine^TM2000、Entranste^TM-D纳米材料载体转染的细胞中荧光蛋白表达也分别达到70％和80％以上，而Entranste^rTM-D纳米材料载体在最高转染效率的剂量下仍然保持80％以上的细胞生存率。RT-PCR进一步证实，三种载体转染细胞均有Math1基因mRNA的表达。结论Entranster^TM—D纳米载体作为一种新型纳米聚合物转染试剂，能成功携带内耳基因治疗关键基因Math1基因转染293T细胞，并表现了低毒、高效性能。