简介：AIM:Totransfectthecatcornealendothelialcells(CECs)withrecombinanthumanβ-nervegrowthfactorgeneadeno-associatedvirus(AAV-β-NGF)andtoobservetheeffectoftheexpressedβ-NGFproteinontheproliferationactivityofcatCECs.METHODS:TheendotheliumofcatcorneawastornunderthemicroscopeandrapidlycultivatedinDulbecco’smodifiedEagle’smedium(DMEM)toformsinglelayerCECsandthepassage2endothelialcellswereusedinthisexperiment.TherecombinanthumanAAV-β-NGFwasconstructed.TherecombinanthumanAAV-β-NGFwastransferredintocatCECsdirectly.Threegroupswereasfollowing:normalCECcontrolgroup,CEC-AAVcontrolgroupandrecombinantCECAAV-β-NGFgroup.Forty-eighthoursaftertransfection,thetotalRNAwasextractedfromtheCECbyTrizol.Theexpressionoftheβ-NGFtargetgenedetectedbyfluorescencequantitativepolymerasechainreaction;proliferationactivityofthetransfectedCECdetectedat48hbyMTTassay;thepercentageofG1cellsamongCECsaftertransfectwasdetectedbyflowcytometrymethod(FCM);cellmorphologywasobservedunderinvertedphasecontrastmicroscope.RESULTS:Thetornendotheliumculturetechniquerapidlycultivatedsinglelayercatcornealendothelialcells.Theself-designedprimersforthetargetgeneandreferencegenewereefficientandspecialconfirmedthroughelectrophoresisanalysisandDNAsequencing.Forty-eighthoursaftertransfect,thehumanβ-NGFgenemRNAdetectedbyfluorescencequantitativepolymerasechainreactionshowedthattherewasnosignificantdifferencebetweennormalCECcontrolgroupandCECAAVcontrolgroup(P>0.05);therewassignificantdifferencebetweentwocontrolgroupsandrecombinantCEC-AAV-β-NGFgroup(P<0.05).MTTassayshowedthattransfectofrecombinantAAV-β-NGFpromotedtheproliferationactivityofcatCEC,whiletherewasnosignificantdifferencebetweennormalCECcontrolgroupandCEC-AAVcontrolgroup(P>0.05).FCMresultshowedthatthepercentageofG1cellsinCECAAV-NGFgroupwas76.8%whi

简介：Leber'scongenitalamaurosis(LCA)andrecentgenetherapyadvancementfortreatinginheritedretinopathieswereextensiveliteraturereviewedusingMEDLINE,PubMedandEMBASE.Adeno-associatedviralvectorswerethemostutilisedvectorsforoculargenetherapy.Conephotoreceptorcellsmightuseanalternatepathwaywhichwasnotreliantoftheretinalpigmentepithelium(RPE)derivedretinoidisomerohydrolase(RPE65)toaccessthe11-cisretinaldehydechromophore.Researcheffortsdedicatedontheprogressionofagene-basedtherapyforthetreatmentofLCA2.Suchgenetherapyapproacheswereextremelysuccessfulincanine,porcineandrodentLCA2models.TherecombinantAAV2.hRPE65v2adenoassociatedvectorcontainedtheRPE65cDNAandwasreplicationdeficient.ItsinvitroinjectionintargetcellsinducedRPE65proteinproduction.Thegenetherapytrialsthatweresofarconductedforinheritedretinopathieshavegeneratedpromisingresults.PhaseIclinicaltrialstocureLCAandchoroideremiademonstratedthatadeno-associatedviralvectorscontainingRPEgenesandphotoreceptorsrespectively,couldbesuccessfullyadministeredtoinheritedretinopathypatients.AphaseIIItrialispresentlyongoingandifsuccessful,itwillleadthewaytoadditionalgenetherapyattemptstocuremonogenic,inheritedretinopathies.

简介：AIM:Tomakecomprehensivemoleculardiagnosisforretinitispigmentosa（RP）patientsinaconsanguineousHanChinesefamilyusingnextgenerationsequencingbasedCapture-NGSscreentechnology.METHODS:Afive-generationHanChinesefamilydiagnosedasnon-syndromicX-linkedrecessiveRP（XLRP）wasrecruited,includingfouraffectedmales,fourobligatefemalecarriersandelevenunaffectedfamilymembers.Capture-NGSwasperformedusingacustomdesignedcapturepanelcovers163knownretinaldiseasegenesincluding47RPgenes,followedbythevalidationofdetectedmutationusingSangersequencinginallrecruitedfamilymembers.RESULTS:Capture-NGSinoneaffected47-year-oldmalerevealsanovelmutation,c.24172418insG:p.E806fs,inexonORF15ofRPGTPaseregulator（RPGR）generesultsinaframeshiftchangethatresultsinaprematurestopcodonandatruncatedproteinproduct.Themutationwasfurthervalidatedinthreeoffouraffectedmalesandtwooffourfemalecarriersbutnotintheotherunaffectedfamilymembers.CONCLUSION:Wehaveidentifiedanovelmutation,c.24172418insG:p.E806fs,inaHanChinesefamilywithXLRP.OurfindingsexpandthemutationspectrumofRPGRandthephenotypicspectrumofXLRPinHanChinesefamilies,andconfirmsCapture-NGScouldbeaneffectiveandeconomicapproachforthecomprehensivemoleculardiagnosisofRP.

简介：AIMTo报告角膜的营养障碍(液晶显示器)与二个变化，R124C和A546D联系了的格子的一个phenotypic变体家谱，在导致贝它的基因(TGFBI).METHODSA详细说明了的转变生长因素，眼睛的检查为一个液晶显示器家庭的所有参加者被参加。从每个参加者的外部血白血球被提取获得DNA。TGFBI基因的所有十七exons的聚合酶链反应(PCR)被执行。产品被定序并且分析。在从proband.RESULTSGenetic分析的右眼睛的渗透的keratoplasty证明proband和所有6个影响个人两个都在codon怀有异质接合的CGC到TGC变化以后，组织学的检查被执行124并且异质接合的GCC到在codon的GAC变化546TGFBI。任何一个100个控制题目和未受影响的家庭成员都不为这二个变化是积极的。眼睛的检查显示了多重refractile在在外部角膜的中央角膜和小小粒的存款的前面的基质的像格子的暗。存款与红显示是的刚果断然被染色在自然淀粉、位于主要观察的前面、中间的stroma.CONCLUSIONWe在TGFBI基因带了二个病原的变化(R124C和A546D)的一个新奇液晶显示器家庭。phenotypic特征与与相应单个变化联系的那些显然不同。结果表明尽管明确的变化是疾病的最重要的基因原因，一些不同修饰词等位基因可以影响显型。

简介：AIM:ToInvestigatethegeneticfindingsandphenotypiccharacteristicsofaChinesefamilywithNorriedisease（ND）.METHODS:MoleculargeneticanalysisandclinicalexaminationswereperformedonaChinesefamilywithND.MutationsintheNorriediseasepseudoglioma（NDP）geneweredetectedbydirectsequencing.Haplotypeswereconstructedandcomparedwiththephenotypesinthefamily.Evolutionarycomparisonsandmutantopenreadingframe（ORF）predictionwerealsoundertaken.RESULTS:TwofamilymemberswithocularmanifestationswerediagnosedwithND.Nosignsofsensorineuralhearinglosswereobservedineitherpatient,whileoneofthemshowedsignsofmildmentalretardation.AnovelheterozygousmutationintheNDPgene,c.-12delAAT,wasdetectedinbothpatients.ThemutationandthemutationbearinghapiotypecosegregatedwiththeNDphenotypeinmalesandwastransmittedfromtheirmothersand/orgrandmothers（Ⅱ:2）.ThemalewithoutNDdidnotharborthemutation.Themutationoccurredatthehighlyconservednucleotides.DRFfinderpredictedthatthemutationwouldleadtotheproductionofatruncatedproteinthatlacksthefirst11N-terminalaminoacids.CONCLUSION:Anovelmutation,c.-12delAATintheNDPgene,wasidentifiedinaChinesefamilywithND.ThismutationcausedNDwithoutobvioussensorineuralhearingloss.Mentaldisorderwasfoundinonebutnottheotherpatients.Theclinicalheterogeneityinthefamilyindicatedthatothergeneticvariantsandepigeneticfactorsmayalsoplayaroleinthediseasepresentation.