简介：Prestinhasbeenidentifiedasamotorproteinresponsibleforouterhaircell(OHC)electromotilityandisexpressedontheOHCsurface.PreviousstudiesrevealedthatOHCelectromotilityanditsassociatednonlinearcapacitanceweremainlylocatedattheOHClateralwallandabsentattheapicalcuticularplateandthebasalnucleusregion.ImmunofluorescentstainingforprestinalsofailedtodemonstrateprestinexpressionattheOHCbasalendsinwhole-mountpreparationoftheorganofCorti.However,therelacksadefinitivedemonstrationofthepatternofprestindistribution.TheOHClateralwallhasatrilaminateorganizationandiscomposedoftheplasmamembrane,corticallattice,andsubsurfacecisternae.Inthisstudy,thelocationofprestinproteinsindissociatedOHCswasexaminedusingimmunofluorescentstainingandconfocalmicroscopy.Wefoundthatprestinwasuniformlyexpressedonthebasolateralsurface,includingthebasalpole.Nostainingwasseenonthecuticularplateandstereocilia.Whenco-stainedwithamembranemarkerdi-8-ANEPPS,prestin-labelingwasfoundtobeintheouterlayeroftheOHClateralwall.Afterseparatingtheplasmamembranefromtheunderlyingsubsurfacecisternaeusingahypotonicextracellularsolution,prestin-labelingwasfoundtobeintheplasmamembrane,notthesubsurfacecisternae.ThedatashowthatprestinisexpressedintheplasmamembraneontheentireOHCbasolateralsurface.

简介：Serioushearingandbalanceimpairmentscanoccurasaresultoflossofhaircellsrelatedtoaging,environmentalstresses(suchasnoisesexposure)orexposuretochemotherapeuticdrugs(suchascisplatinandaminoglycosideantibiotics).Becausealargeportionofhearingimpairmentinvolveslossofhaircells,regenerationorreplacementofthesecellsisapossiblealternativetoprostheticdevices1.Researchershavepaidhotattentiontothestudyofinnerearhaircellregenerationforhearingrestorationformanyyears.Besidestheimportantfunctionofhaircells,therearemanyothercontributorsessentialfornormalhearing.Haircellregenerationisoneoftheessentialstepsfortotalhearingrecovery,butnottheonlyone.Inthisarticle,wewillreviewtheprogressesinresearchonhearingloss,stemcellapplicationandhearingrestoration.

简介：ObjectivesLangerhans'Cellhistiocytosis(LCH)isararedisease,whichremainspoorlyunderstoodandwhosecellularoriginremainsunknown.ToincreaseunderstandingoftemporalboneLCH,itisnecessarytostudyrecentadvancesinthediagnosisandtreatmentofthisdisease.MethodsThelongterm(5to30years)resultsof21temporalboneLCHcasestreatedbetween1973and2003werereviewed.Surgery,radiotherapy,pharmacologictherapyoracombinationofthesetreatmentswereemployedinthesecases.ResultsEighteenpatientswerecured(18/21,85%).Sixpatientsdevelopedresidualdiabetesinsipidus(DI)anddwarfism(28%).Threepatientsdied(14%).ConclusionsTheAlessiclassificationsystemforLCHbasedontheextentofdiseaseaccuratelypredictsprognosisandisausefulguideinselectingtreatmentmethodologies.X-ray,computedtomographyandmagneticresonanceimaginghaveprovedusefulindefiningtheextentofosseousandsofttissuediseases.DiagnosisofLCHisbasedonclinicalpresentations,radiographicfindingsandhistopathologicalresults.Surgeryandradiotherapyarethemaintreatmentmodalities.Pharmacologictherapyshouldbeusedinpatientswithaggressive,disseminate,andrefractorylesions.LCHhasapredilectionforchildrenandprognosisdependsonageandextentofvitalorganinvolvement.

简介：Cell-celljunctionsinthecochleaarehighlycomplexandwellorganized.Theroleofthesejunctionsistomaintainstructuralandfunctionalintegrityofthecochlea.Inthisreview,wedescribeclassificationofcelljunction-associatedproteinsidentifiedwithinthecochleaandprovideabriefoverviewofthefunctionoftheseproteinsinadherentjunctions,gapjunctionsandtightjunctions.

简介：ForwardMaskingTemporalaudiotoryresolutionistheabilityoftheauditorysystemtoresolveauditorysignalsinthetimedomain.Forwardmaskingisameansofstudyingtemporalresolutionwhereonetone,theprobe,ismaskedbyaprecedingtone,themasker.Forwardmaskingis

简介：Theelectricallyevokedsomaticmotilityofouterhaircells(OHC),brieflytermedOHCelectromotility,playsacrucialroleincochlearamplificationthatunderliestheremarkablyhighsensitivityandfrequencyselectivityofthemammalianhearing.AccompanyingOHCelectromotilityisavoltage-dependentgatingchargemovementwithinthecelllateralmembrane,manifestedasameasurablenonlinearcapacitance(NLC)inOHCs.TheelectromotilityandNLCofOHCsarehighlycorrelatedbysharingacommonmolecularsubstrate,themotorproteinprestin.Inthisstudy,wesystematicallycharacterizedthequantitativerelationshipbetweenOHCelectromotilityandNLCintheirvoltagedependencesforthepurposeoffurtherunderstandingtheelectromechanicaltransductioninOHCs.TheresultsdemonstratedthatthetwopossessdifferingvoltagedependenceswiththeV1/2ofelectromotilityconsistentlybeing～20mVdepolarizedincomparisonwiththatofNLCalthoughtheirslopefactorsαarestatisticallyidentical.FurtherinvestigationsshowedthattheinitialstateofOHCsinfluencesthevoltagedependenceofelectromotilitybutnotthatofNLC,indicatingthatsomebiophysicalfactorsotherthanthemotorproteinperseareinvolvedintheprocessofOHClengthchanges.Weproposedthatthecytoskeletalspectrin-actinframeworkunderneaththeOHCplasmamembraneandthecell'sturgorarethetwomostprobablefactorsthatcausethevoltage-dependencediscrepancybetweenOHCelectromotilityandNLC.

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：ObjectiveTounderstandthemechanismofnoiseexposureinducedouterhaircells(OHCs)deathpathways.MethodsThirtytwoguineapigswereusedinthisstudy.Theanimalswereeitherexposedfor4h/daytobroadbandnoiseat122dBSPL(A-weighted)for2consecutivedaysorperfusedwithMNNG.Afterauditorytest,thecochleaeofanimalsweredissected.Propidiumiodide(PI),aDNAintercalatingfluorescentprobe,wasusedtotracemorphologicalchangesinOHCnuclei.F-actinstainingwasusedtodeterminemissingOHCs.Caspase-3wasdetectedinlivingorganofCortiwholemountsusingthefluorescentprobe.ThesinglestrandDNA(ssDNA)inapoptoticOHCsinguineapigsandapoptosisinducingfactor(AIF)inhaircellsinguineapigswereexaminedbyimmunohistologymethod.WholemountsoforganofCortiwereprepared.Morphologicalandfluorescentchangeswereexaminedunderaconfocalmicroscope.Results(1)Bothapoptoticandnecrotichaircellsappearedfollowingnoiseexposure.(2)NoiseexposureinducedsinglestrandDNAinapoptoticOHCsbutnotinthenormalOHCs.(3)EitherafternoiseexposureorafterMNNGperfusion,apoptoticOHCswerefeaturedbynuclearcondensationorfragmentationwithcaspase-3activation,whereasnecroticOHCswerecharacterizedbynuclearswellingwithoutcaspase-3activation.(4)InnormalorganofCorti,AIFwaslocatedinthemitochondriaareas.Afternoiseexposure,AIFwastranslocatedfrommitochondriainapoptoticandnecroticOHCs.ConclusionThesefindingsindicatethatnoiseexposuredamagesDNAintheOHC,whichtriggersactionofCaspase-3.Subsequently,AIFistranslocatedtothenucleus,leadingtoDNAdamageandOHCsdeath.

简介：BackgroundNormally,fewimmunocompetentcellarepresentintheendolymphaticsac(ES).Duringanactiveimmuneresponseintheinnerear,largeamountofinflammatorycells,includingimmunocompetentcells,areseenintheES.ThecurrentstudyaimedatassessingcellularproliferationwithintheESduringinducedimmuneresponseintheinnerear.MethodsFifteenhealthy,femaleSDratsweresensitizedsystemicallywithkeyholelimpethemocyanin(KLH),followedbylocalinoculationinthecochleathroughbasalturnfenestrationwiththesameantigen.OnDays3,7and14followinginoculation,theanimalwassacrificedafterintraperitonealadministrationof5-bromo-2'-deoxyuridine(BrdUrd),andthetemporalboneharvested.Followingdecalcification,infiltrationbyBrdUrd-andIgG-positivecellsintheESwasstudiedonfrozensectionswithH&Eandimmunohistochemicalstaining.ResultsDuringthesecondaryimmuneresponseintheinnerearagainstT-dependentantigens,thereisincreasedcellularproliferationintheES.Theproliferatedcellsmaydifferentiateintoimmunocompetentcellsatthesamelocation.ConclusionsThesefindingsindicatethattheESplaysanimportantroleinimmuneresponseofinnerear.

简介：Cisplatinbelongstoplatinum-baseddrugsandiswidelyusedincancerchemotherapy.Ototoxicityisoneofthemajordoselimitingside-effectsofcisplatin.Fortoxicitytooccurcisplatinmustfirstbetransportedfromthebloodstreamintocochlearcells.Threecoppertransportersareconsideredpathwaysforregulatingtheuptakeandtranslocationofcisplatinintocells:Ctr1,ATP7AandATP7B.Ourrecentstudywithcochlearorganotypicculturesshowsthatcochlearhaircellscanbedestroyedbycisplatinatlowconcentrationsfrom10μmto100μn.However,highdosesofcisplatincannotdamagehaircells,maybeduetointrinsicfeedbackreactionsthatincreaseexportofplatinumbyATP7Bwhentheplatinumconcentrationishighinextracellularspace.Cimitidineisaspecificcoppertransporterinhibitorthatcanblocktheentranceofcopperandplatinum,andmaypreventcisplatin-inducedcochlearhaircellinjury.Toevaluatethishypothesis,wetreatedcochlearorganotypiccultureswithcisplatin(10μmor50μm)alone,orcisplatincombinedwithcimitidineatconcentrationsrangingfrom10-2000μmfor48hours.cisplatinat10μmdamagedabout20%haircells.Incontrast,whencimitidine(10μm,100μmand2000μm)wasaddedtotheculture,near100%cochlearhaircellsurvived.Athigherconcentration(50μm),cisplatindestroyedabout80%ofcochlearhaircells.However,100μmcimitidinerescuedabout50%haircellsfromcisplatindamage,and2000μmcimitidineprotectedabout80%haircells.ThedataofwesternblotshowedthatCTR1andATP7Bexpressionswereincreasedincisplatintreatedcochleartissue,butcimitidinesignificantlyreducedCTR1andATP7B.Inaddition,ATP7Aexpressionwasdepressedalittleaftercisplatintreatment.ConsideringthatCtr1isinvolvedincopperandplatinuminflux,buttheATP7AandATP7Barecopperexporttransporters,theresultssuggestthatcimitidinecaneffectivelyblocktheentrancebycoppertransportersandstoptheinfluxofcisplatin.

简介：ObjectiveTopresentanexperimentalmethodthatallowsisolationofgreaterepithelialridge(GER)andlesserepithelialridge(LER)cellsfrompostnatalratcochleaeusingacombinatorialapproachofenzymaticdigestionandmechanicalseparationandtoinvestigatearetrovirus-mediatedgenetransfertechniqueforitspossibleutilityinimmortalizationoftheGERandLERcelllines,inanefforttoestablishaninvitromodelsystemofhaircelldifferentiation.MethodsGERandLERcellsweredissectedfrompostnatalratcochleaeandimmortalizedbytransferringtheSV40largeTantigenusingaretrovirus.Theestablishedcelllineswereconfirmedthroughmor-phologyobservation,immunnocytochemicalstainingandRT-PCRanalysis.TheHath1genewastransferredintothecelllinesusingadenovirus-mediatedtechniquestoexploretheirpotentialtodifferentiateintohaircells.ResultsTheestablishedcelllineswerestablymaintainedformorethan20passagesanddisplayedmanyfeaturessimilartoprimaryGERandLERcells.Theygrewinpatchesandassumedapolygonalmorphology.ImmunostainingshowedlabelingbySV40largeTantigenandIslet1(aspecificmarkerforGERandLER).AllpassagesofthecelllinesexpressedSV40largeTantigenonRT-PCRanalysis.Thecellsalsoshowedthecapabilitytodifferentiateintohaircell-likecellswhenforcedtoexpressHath1.ConclusionRetrovirus-mediatedgenetransfercanbeusedinestablishingimmortalizedprogenitorhaircelllinesinnewbornrat,whichmayprovideaninvaluablesystemforstudyinghaircelldifferentiationandregenerationfornewtreatmentofsensoryhearinglosscausedbyhaircellloss.

简介：Cisplatin,awidelyusedanticancerdrug,damageshaircellsincochlearorganotypicculturesatlowdoses,butparadoxicallycauseslittledamageathighdosesresultinginaU-shapeddose-responsefunction.Todetermineifthecisplatindose-responsefunctionforvestibularhaircellsfollowsasimilarpattern,wetreatedvestibularorganotypiccultureswithdosesofcisplatinrangingfrom10to1000μM.Vestibularhaircelllesionsprogressivelyincreasedasthedoseofcisplatinincreasedwithmaximumdamageoccurringaround50–100μM,butthelesionsprogressivelydecreasedathigherdosesresultinginlittlehaircelllossat1000μM.TheU-shapeddoseresponsefunctionforcisplatin-treatedvestibularhaircellsincultureappearstoberegulatedbycoppertransporters,Ctr1,ATP7AandATP7B,thatdose-dependentlyregulatetheuptake,sequestrationandextrusionofcisplatin.

简介：ObjectiveAdipose-derivedstemcells(ADSCs)aresuggestedtopossessahighlyplasticabilitytodifferentiateintoseveralspecificcelltypesinadditiontoadipocytelineages,includinggermlayertissue-specificcelllineagessuchaschondrocyte,myocyte,neuronal,andosteoblastlineages.TheaimofthisstudyistoestablishaninvitroculturetechniqueforADSCsinanadultguineapigmodelthatfacilitatetheirdifferentiationintohaircell-likecells.MaterialsandMethodsCellsfrominguinalfatpadsinadultguineapigswereculturedwithβ-mercaptoethanol,RA,Forskorin,Heregulin,bFGF,BDNFandEGF.Cellulardifferentiationwasexaminedusingimmunocytochemistrytechniques.ResultsTheADSCsdemonstratedhaircellimmunophenotypeswithexpressionofepitopesofthehaircellmarkerproteinmyosinⅦa.ConclusionADSCsfromadultguineapigadiposetissuecandifferentiateintohaircell-likecellswhenculturedinvitro.ADSCsmayserveasseedcellsfortissueengineering.

简介：Togaininsightsintotheototoxiceffectsofaminoglycosideantibiotics(AmAn)anddelayedperipheralganglionneurondeathintheinnerear.experimentalanimalmodelswerewidelyusedwithseveraldifferentapproachesincludingAmAnsystemicinjections,combinationtreatmentofAmAnanddiuretics,orlocalapplicationofAmAn.Intheseapproaches,systemicAmAntreatmentaloneusuallycausesincompletedamagetohaircellsintheinnerear.Co-administrationofdiureticandAmAncancompletelydestroythecochlearhaircells,butitisimpossibletodamagethevestibularsystem.OnlytheapproachofAmAnlocalapplicationcanselectivelyeliminatemostsensoryhaircellsintheinnerear.Therefore,AmAnlocalapplicationismoresuitableforstudiesforcompletehaircelldestructionsincochlearandvestibularsystemandthefollowingdelayedperipheralganglionneurondeath.Incurrentstudies,guineapigswereunilaterallytreatedwithahighconcentrationofgentamicin(GM,40nig/ml)throughthetympanicmembraneintothemiddleearcavity.AuditoryfunctionsandvestibularfunctionsweremeasuredbeforeandafterGMtreatment.Thelossofhaircellsanddelayeddegenerationofganglionneuronsinbothcochlearandvestibularsystemwerequantified30daysor60daysaftertreatment.TheresultsshowedthatbothauditoryandvestibularfunctionswerecompletelyabolishedafterGMtreatment.Thesensoryhaircellsweretotallymissinginthecochlea,andseverelydestroyedinvestibularend-organs.Thedelayedspiralganglionneurondeath60daysafterthedeafeningprocedurewasover50%.However,noobviouspathologicalchangeswereobservedinvestibularganglionneurons60dayspost-treatment.Theseresultsindicatedthatahighconcentrationofgentamycindeliveredtothemiddleearcavitycandestroymostsensoryhaircellsintheinnerearthatsubsequentlycausesthedelayedspiralganglionneurondegeneration.Thismodelmightbeusefulforstudiesofhaircellregenerations,delayeddegenerationofperipheralauditoryne