简介：ObjectiveThestudyistoidentifythecarrierrateofcommondeafnessmutationinChinesepregnantwomenviadetectingdeafnessgenemutationswithgenechip.MethodsThepregnantwomeninobstetricclinicwithouthearingimpairmentandhearingdisordersfamilyhistorywereselected.Theinformedconsentwassigned.PeripheralbloodwastakentoextractgenomicDNA.Applicationofgeneticdeafnessgenechipfordetecting9mutationalhotspotofthemostcommon4Chinesedeafnessgenes,namelyGJB2(35delG,176del16bp,235delC,299delAT),GJB3(C538T),SLC26A4(IVS72A>G,A2168G)andmitochondrialDNA12SrRNA(A1555G,C1494T).Furthergenetictestingwereprovidedtothespousesandnewbornsofthescreenedcarriers.ResultsPeripheralbloodof430pregnantwomenweredetected,detectionofdeafnessgenemutationcarriersin24cases(4.2%),including13casesoftheGJB2heterozygousmutation,3casesofSLC26A4heterozygousmutation,1casesofGJB3heterozygousmutation,and1caseofmitochondrial12SrRNAmutation.18spousesand17newbornstookfurthergenetictests,and6newbornsinheritedthemutationfromtheirmother.ConclusionThecommondeafnessgenesmutationhasahighcarrierrateinpregnantwomengroup,235delCandIVS7-2A>Gheterozygousmutationsarecommon.

简介：Object:ToidentifytranscriptvariantsandexpressionpatternsofporcineMitf.Materialsandmethods:ApairwiseBLASTsearchatNCBIdatabasewasperformedtodeducethestructureofporcineMitfgene.Subsequently,50RACEandfluorescentquantitativeRT-PCRwereusedtoanalyzetheexpressionpatternofporcineMitfindifferenttissues.Results:FourtranscriptvariantsofporcineMitf,MITF-A,MITF-H,MITF-MandMITF-SUSwereidentified,allsharinghighhomologywiththoseinhumans,exceptMitf-SUS.Conclusion:ThesequenceofporcineMitfappearhighlyhomologoustohumanMITF.However,only4transcriptvariantsofporcineMitfwereidentifiedintheseminipigs,lessthanthe9transcriptvariantsinhumanMITF.

简介：Objective:Tounderstandthecrucialroleoftheklothogeneinhearingdevelopmentinmousemodels.Methods:PCRwasusedtoidentifyCBAmicewithdifferentgenotypes,i.e.WT,heterozygous(klotho+/-)orhomozygous(klotho-/-).Micephenotypeandweightwererecordedpostnatal25days(P-25)andauditorybrainstemresponses(ABR)wereusedtodetermineauditoryfunctionatP-60.Results:klotho-/-micetendedtohavesmallersize,lighterweightandhigherABRthresholdsatP-60,showingearlyonsetage-relatedhearingloss(ARHL).Conclusion:Heterozygousandhomozygousklothodeficientmiceexhibitdifferentdegreesofhearinglossatyoungage,withhomozygousmice(klotho-/-)showingmoreseverehearingloss.OurresultsindicatethatpersistedexpressionofklothoproteinintheinnerearmaypotentiallydelaytheonsetofARHLandplayanimportantroleintheprotectionofauditoryfunction.

简介：Cisplatindamagescochlearhaircellsandspiralganglionneuronsthroughcelldeathsignalingpathwaysthatarenotfullyunderstood.Weusedfocusedapoptosisgenemicroarraystostudyearlychangesingeneexpres-sionincochlearculturesfromP3neonatalratstreatedwithcisplatin(0.2mM).After12hoursofcisplatintreat-ment,morethan50%ofthe96genesonthearrayshowedasignificantdecreaseinexpression,consistentwithwidespreadcelldeath.However,after3hoursofcisplatintreatment,10genesshowedsignificantincreaseinex-pressionintotalcochleartissue.Inexperimentswithsubsetsofcochleartissues,at3h,cisplatininducedincreasedexpressionof12genesinthecochlearsensoryepithelium(basilarmembrane)and11genesinthespiralganglion(tissueofRosenthal'scanal,containingthespiralganglion).Theseincludedpro-andanti-apoptoticgenesin-volvedinthep53signalingpathway,TNFreceptorfamily,NF-kappaBpathway,deathdomainfamily,deatheffec-tordomainfamily,Bcl-2family,CARDfamily,TRAFfamily,andGTPsignaltransduction.Althoughthechangesingeneexpressionshowedanoverlapbetweenbasilarmembraneandspiralganglion,otherchanges,whichmayreflecttheuniqueresponseofeachtissue,werealsoobserved.Pifithrin-αblockedcisplatin-inducedup-regulationofgenesinthep53signalingpathwaywhenassayedbybothsuperarrayandrealtimePCR.Thedataaddtoourunderstandingoftheinvolvementofp53incisplatin-inducedototoxicityandotoprotection,conferredbythep53inhibitorPifithrin-α.

简介：FragileXsyndromeisthemostcommonformofinheritedmentalretardationaffectingupto1in4000individuals.ThesyndromeisinducedbyamutationintheFMR1gene,causingadeficiencyinitsgeneby-productFMRP.ImpairmentinthenormalfunctioningofFMRPleadstolearningandmemorydeficitsandheightenedsensitivitytosensorystimuli,includingsound(hyperacusis).ThemolecularbasisoffragileXsyndromeisthoroughlyunderstood;however,theneuralmechanismsunderlyinghyperacusishavenotyetbeendetermined.Astheinferiorcolliculus(IC)istheprincipalmidbrainnucleusoftheauditorypathway,thecurrentstudyaddressesthequestionsunderlyingtheneuralmechanismofhyperacusiswithintheICoffragileXmice.AcuteexperimentswereperformedinwhichelectrophysiologicalrecordingsoftheICinFMR1-KOandWTmiceweremeasured.ResultsshowedthatQ-valuesforWTweresignificantlylargerthanthatofFMR-1KOmice,indicatingthatWTmiceexhibitsharpertuningcurvesthanFMR1-KOmice.WealsofoundtheratioofthemonotonicneuronsintheKOmicewasmuchhigherthantheWTmice.TheseresultssuggestthatlackofFMRPintheauditorysystemaffectsthedevelopmentalmaturationandfunctionofstructureswithintheauditorypathway,andinthiscasespecificallytheIC.ThedysfunctionobservedwithintheauditoryneuralpathwayandinparticulartheICmayberelatedtotheincreasedsusceptibilitytosoundasseeninindividualswithfragileXsyndrome.OurstudymayhelponunderstandingthemechanismsofthefragileXsyndromeandhyperacusis.

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：Objective:Congenitalauditoryneuropathy(AN)affectshearingandspeechdevelopment.ThedegreeofhearingdifficultyincongenitalANvariesasafunctionofpathologyattheinnerearhaircell(IHC)synapsesortheauditorynerve.WereportacaseofaChinesegirlwithANrevealedbyOTOF(otoferlin)genemutationanalysiswhohadonlyamildhearingloss.Patient:A13-year-oldChinesegirlwasdiagnosedashavingcongenitalANonthebasisofOTOFgenemutationanalysis.Shemanifestamildsensorineuralhearinglosswith50%maximummonosyllablespeechdiscriminationrate,normalDPOAEs(distortionproductotoacousticemissions)beyondambientnoiselevels,onlySPs(summatingpotentials)evokedduringECoG(electrocochleography)andabsentABRs(auditoryevokedbrainstemresponses)bilaterallytoclickspresentedat100dBnHL.Shewasabletoeffectivelycommunicatewithothersbyspeechreadingowingtohermildhearingloss.Moreover,bilateralhearingaidshelpedhertocommunicate.Conclusions:OurpatientwasdemonstratedtohaveamutationontheOTOFgene.Nevertheless,shewasabletocommunicateusingauditoryvisualspeechreadinginspiteofamildauditorythresholdelevationprobablyduetopartialpathologyattheIHCsynapsesorintheauditorynerve.

简介：Microphthalmia-associatedtranscriptionfactor(MITF)controlsmelanocytesurvivalanddifferentiationthroughdirectlyregulatingtheexpressionofthetyrosinase(TYR)andtyrosinase-relatedproteins1and2(TYRP1andTYRP2)genes.MITFmutationshavebeenreportedtoresultinanabnormalmelanocytedevel-opmentandleadtoWaardenburgsyndrometype2(WS2),characterizedbyvariabledegreesofsensorineu-ralhearinglossandpatchyregionaldistributionofhypopigmentation.Recently,MITFwasalsoindicatedasacausativegeneforamoreseveresyndrome,theTietzSyndrome(TS),characterizedbygeneralizedhy-popigmentationandcompletehearingloss.However,fewfunctionalstudieshavebeenperformedtocom-parethediseases-causingmutations.Here,weanalyzedtheinvitroactivityoftworecentidentifiedWS2-as-sociatedmutation(p.R217Iandp.T192fsX18)andoneTS-associatedmutationp.N210K.TheR217IMITFretainedpartialactivity,normalDNA-bindingabilityandnucleardistribution,whereastheT192fsX18MITFfailedtoactivateTYRpromoterduetolossofDNA-bindingactivity,andaberrantsubcellularlocalization.TheaberrantsubcellularlocalizationofT192fsX18MITFmaybecausedbydeletionofaputativenuclearlocalizationsignal(NLS)ataa213-218(ERRRRF).Indeed,MITFwithdeletionoftheNLSfragmentfailedtotranslocateintothenucleusandactivatedtheTYRpromoter.TaggingthisNLStoGFPpromotedthegreenfluorescenceprotein(GFP)translocatedintothenucleus.ThesurprisingfindingofourstudyisthataTS-as-sociatedMITFmutation,N210K,showedcomparableinvitroactivityasWT.Thus,thepossibleinvolve-mentofMITFinTSanditsunderlyingmechanismsstillneedfurtherinvestigation.

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：Noise-inducedhearinglossisacommoncauseofacquiredhearinglossintheadultpopulation.Acousticoverstimulationcausescochleardamagethroughmechanicalstresstothetissue.Consequently,complexmolec-ularchangesareinitiated,andthesechangesleadtomorphologicalandbiologicalalterationsinthecochlea,whichinturncompromisethecochlearfunctionandcausehearingloss.Inthepast10years,therehavebeensignificantadvancesinourunderstandingofthemolecularmechanismsofnoise-inducedhearingloss.Theseadvancesareattributed,inpart,tothedevelopmentofhigh-throughputtechnologiesfortheglobalanalysesofmolecularchanges.Inthisreview,webrieflydescribethenewlydevelopedmethodsforinvestigatingthemo-lecularresponsesofthecochleatoacoustictraumaandtheknowledgegeneratedfromthesestudies.Wealsodiscussthestrengthsandlimitationsofeachtechniqueandthemajorchallengestoinvestigatecochleardegen-erationfollowingacousticinjury.

简介：ObjectiveTostudyexpressionofadenoviral-mediatedHath1-EGFPgeneintheguineapigcochleaaftertransferthroughintactroundwindowmembrane(RWM),andtoassessitseffectsonhearing.MethodsTwentyadultguineapigswereused,ofwhich:12weresurgicallyinoculatedwithAd-Hath1-EGFPinthebonygrooveofroundwindowniche,and8withartificialperilymph.Auditorybrainstemresponse(ABR)thresholdsweredeterminedinallanimalsbeforeand5daysaftersurgery.Onpost-surgeryday5andday14,animalsweresacrificedandwholemountsofcochleaandfrozensectionswereexamined.ResultsABRtestsshowednosignificantchangeofhearingafterthesurgery.StrongfluorescencestaininginthecochleaewasseeninAd-Hath1-EGFPgroups.Thehighestlevelsofgeneexpressionwereseeninthepost-surgeryday5groupwithlittledecreaseonpost-surgeryday14.Thecontralateralcochleaandthoseinthecontrolgroupswerefreeoffluorescencestaining.ConclusionThetransgenicHath1-EGFPcanbeeffectivelydeliveredintotheinnerearthroughintactRWM,inanatraumaticmanner.