簡介：244ARCHPATHOLLABMEDVOL132,FEBRUARY2008MYCOBACTERIUMTUBERCULOSIS,GENETICELSAYEDZAKISAMIRABOUELHASSAN,MD●CONTEXTDIAGNOSTICDETECTIONOFTUBERCULOSISTBHASIMPROVEDCONSIDERABLYAVAILABLE,STANDARDIZED,NUCLEICACID–BASEDAMPLIFICATIONTECHNIQUESHAVEBEENSHOWNTOYIELDRELIABLERESULTSWITHIN4TO7HOURSOFSAMPLEPROCESSINGOBJECTIVETOSTUDYTHEDIAGNOSTICPERFORMANCEOFGENPROBE’STECHNIQUEFORDIRECTDETECTIONOFMYCOBACTERIUMTUBERCULOSISINCOMPARISONWITHBACTEC460TBCULTUREFORBOTHPOSITIVEANDNEGATIVEZIEHLNEELSENSMEARSINEGYPTIANCHILDRENATRISKFORTBINFECTIONDESIGNWEPROSPECTIVELYEVALUATED50CHILDRENFROMFAMILIESWITHAPOSITIVEHISTORYOFTBALLPATIENTSWEREREFERREDFROMOUTPATIENTCLINICSOFTHEMANSOURAUNIVERSITYCHILDREN’SHOSPITAL,EGYPTTHECHILDRENHADAPOSITIVETUBERCULINSKINTESTWITHANINDURATIONDIAMETEROFMORETHAN10MMANDHADSCARSFROMABACILLECALMETTEGUE′RINVACCINATIONWITHINTHEPAST2YEARSTHREECONSECUTIVESPUTUMSAMPLESWERETAKENFROMEACHPATIENTTHESAMPLESWEREEXAMINEDTODETECTMTUBERCULOSISBYMEANSOFTHEGENPROBETECHNIQUE,DIRECTSMEARMICROSCOPY,ANDBACTERIALCULTUREBYBACTEC460TBRESULTSOFTHE50CASES,3060HADSPUTUMSAMPLESTHATWEREPOSITIVEFORTBBYBACTEC460TBCULTURE,AND29CASES58WEREPOSITIVEBYTHEGENPROBETECHNIQUESENSITIVITYANDSPECIFICITYOFZIEHLNEELSENSMEARSWAS833AND100,RESPECTIVELY,WITHOVERALLACCURACYOF90SENSITIVITYANDSPECIFICITYOFTHEGENPROBETECHNIQUEWERE967AND100,RESPECTIVELY,WITHOVERALLACCURACYOF98CONCLUSIONSTHERESULTSOFTHISSTUDYSUGGESTTHATTHEGENPROBETECHNIQUEISANACCURATEMETHODFORRAPIDDETECTIONOFMTUBERCULOSISCOMPLEXESINRESPIRATORYSAMPLESFROMCHILDRENATRISKFORTBITCANBEUSEDFORDIAGNOSISOFSMEARNEGATIVECASESTHATARESUSPECTFORTBARCHPATHOLLABMED2008132244–247CHILDHOODTUBERCULOSISTBHASITSHIGHESTINCIDENCEAMONGCHILDRENINCONTACTWITHBACILLIFEROUSADULTS1,2THEPRESENTSTUDYWASMOTIVATEDBYTHEINCREASEINTHENUMBEROFTBCASESTHATAREBEINGOBSERVEDINCHILDRENTRADITIONALLABORATORYDIAGNOSISOFMYCOBACTERIALINFECTIONSBYCULTUREUSUALLYREQUIRES2TO8WEEKSTHERECENTINCREASEINNEWCASESOFTBHASSHOWNTHENEEDFORRAPID,SPECIFIC,DIAGNOSTICASSAYSFORMYCOBACTERIUMTUBERCULOSIS3WITHTHEDEVELOPMENTOFNOVELTECHNIQUESINMOLECULARBIOLOGY,THISDELAYMIGHTBESHORTENEDMOSTOFTHENUCLEICACIDAMPLIFICATIONASSAYSARERAPIDANDSPECIFIC4DESPITETHEIRTHEORETICALABILITYTODETECTEVENASINGLEMYCOBACTERIALCELL,NUCLEICACIDAMPLIFICATIONTESTSNAATSARENOTSUFFICIENTLYRELIABLETOREPLACECONVENTIONALDIAGNOSTICMETHODSFORDETECTINGTBINHERENTTESTCHARACTERISTICSANDERRORSINTESTINGPROCEDURESMAYACACCEPTEDFORPUBLICATIONSEPTEMBER17,2007FROMTHEDEPARTMENTSOFCLINICALPATHOLOGYDRELSAYEDZAKIANDPEDIATRICSDRABOUELHASSAN,FACULTYOFMEDICINE,MANSOURAUNIVERSITYEGYPTTHEAUTHORSHAVENORELEVANTFINANCIALINTERESTINTHEPRODUCTSORCOMPANIESDESCRIBEDINTHISARTICLEREPRINTSMAYSAAELSAYEDZAKI,MD,EGYPTMANSOURAUNIVERSITY,FACULTYOFMEDICINE,DEPARTMENTOFPATHOLOGY,MANSOURA65EGYPTEMAILMAY?S65HOTMAILCOMCOUNTFORTHEIRINACCURACY5FURTHERMORE,THEPRESENCEINRESPIRATORYSECRETIONSOFENZYMESCAPABLEOFINHIBITINGAMPLIFICATIONREACTIONSACCOUNTSFORANADDITIONAL3TO25OFFALSENEGATIVERESULTS6ONTHEOTHERHAND,FALSEPOSITIVERESULTSARISEMOSTOFTENFROMCONTAMINATIONOFNEGATIVESAMPLESWITHEITHERORGANISMSORTARGETDNAFROMSAMPLESCONTAININGLARGENUMBERSOFMYCOBACTERIAORFROMAMPLICONSCONTAMINATINGTHELABORATORYROOM5,6TOOVERCOMETHESEPROBLEMS,AUTOMATEDCOMMERCIALSYSTEMSWEREDEVELOPEDTHATWEREMADEMOREROBUSTBYTHEUSEOFSTANDARDIZEDPROCEDURESANDREAGENTSFORSAMPLEPROCESSING,AMPLIFICATION,ANDDETECTIONVARIOUSAUTOMATEDSYSTEMS,BASEDONAMPLIFICATIONANDDETECTIONTECHNIQUES,HAVEBEENDEVISEDFORTHEDETECTIONOFMTUBERCULOSISINCLINICALSAMPLESTHESYSTEMSINCLUDETHEPOLYMERASECHAINREACTION–BASEDCOBASAMPLICORMYCOBACTERIUMSYSTEMROCHEDIAGNOSTICS,BASEL,SWITZERLAND7THETRANSCRIPTIONMEDIATED,AMPLIFICATIONBASEDAMPLIFIEDMYCOBACTERIUMTUBERCULOSISDIRECTTESTAMTDGENPROBE,INC,SANDIEGO,CALIF8THESTRANDDISPLACEMENTAMPLIFICATIONBASEDBDPROBETECETSYSTEMBECTONDICKINSONANDCOMPANY,FRANKLINLAKES,NJ9ANDTHELIGASECHAINREACTION–BASEDABBOTTLCXMTUBERCULOSISASSAYSYSTEMABBOTTLABORATORIES,NORTHCHICAGO,246ARCHPATHOLLABMEDVOL132,FEBRUARY2008MYCOBACTERIUMTUBERCULOSIS,GENETICELSAYEDZAKIHOWEVER,THELATTERLACKSSENSITIVITYANDISUNABLETODISTINGUISHTUBERCLEBACILLIFROMOTHERMYCOBACTERIA22FORRESULTSOFAMTDCOMPAREDWITHCULTURE,29CASESWEREGENPROBEPOSITIVEANDCULTUREPOSITIVE,AND1CASEWASNEGATIVEBYGENPROBEANDCULTUREPOSITIVEFOURSAMPLESOFAFBSMEARSWEREPOSITIVEBYCULTUREANDGENPROBEGENERALLY,DIFFERENCESBETWEENCUTOFFVALUESOFPOSITIVEANDNEGATIVECONTROLSANDSPECIMENSWEREBROADENOUGHTOPERMITEASYDISCRIMINATIONNEGATIVERESULTSOBTAINEDBYAMTDFORCULTUREPOSITIVESPECIMENSMAYBEEXPLAINEDBYUNEQUALDISTRIBUTIONOFASMALLNUMBEROFMYCOBACTERIA23ITISCLEARTHATTHEGENPROBETECHNIQUECANBEUSEDFORTHECONFIRMATIONOFTBINAPERCENTAGEOFTHOSEPROVIDINGAFBSAMPLESASIMILARCONCLUSIONWASREPORTEDBYGRECOETAL23FORMOSTAUTOMATEDSYSTEMSTHEIMPACTOFTHENAATSONPATIENTOUTCOMEVARIESBASEDONTHERESULTOFTHEAFBSMEARINSMEARPOSITIVEPATIENTS,PUBLICHEALTHANDHOSPITALINFECTIONCONTROLRESOURCESAREPREDOMINANTLYAFFECTEDTHEPOTENTIALFORINFLUENCINGPATIENTOUTCOMEISMUCHGREATERWHENTHEAFBSMEARISNEGATIVEINSMEARNEGATIVEPATIENTS,THENAATCOULDPROVIDEMORERAPIDDIAGNOSISOFTBANDSUBSEQUENTINITIATIONOFTHERAPYTHISWOULDELIMINATETHENEEDFORINVASIVEDIAGNOSTICPROC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簡介：LOSSOFEPHRINA5FUNCTIONDISRUPTSLENSFIBERCELLPACKINGANDLEADSTOCATARACTMARGARETACOOPERA,1,ALEXANDERISONA,1,DANIELKOMLOSB,YUHAISUNA,NORMANJKLEIMANC,ANDRENPINGZHOUA,2ADEPARTMENTOFCHEMICALBIOLOGY,SUSANLEHMANCULLMANLABORATORYFORCANCERRESEARCH,ERNESTMARIOSCHOOLOFPHARMACY,RUTGERSUNIVERSITY,PISCATAWAY,NJ08854BDEPARTMENTOFNEUROSCIENCEANDCELLBIOLOGY,ROBERTWOODJOHNSONMEDICALSCHOOL,PISCATAWAY,NJ08854ANDCDEPARTMENTOFENVIRONMENTALHEALTHSCIENCES,MAILMANSCHOOLOFPUBLICHEALTH,COLUMBIAUNIVERSITY,NEWYORK,NY10032COMMUNICATEDBYALLANHCONNEY,RUTGERS,THESTATEUNIVERSITYOFNEWJERSEY,PISCATAWAY,NJ,SEPTEMBER9,2008RECEIVEDFORREVIEWJUNE16,2008CELL–CELLINTERACTIONSORGANIZELENSFIBERCELLSINTOHIGHLYORDEREDSTRUCTURESTOMAINTAINTRANSPARENCYHOWEVER,SIGNALSREGULATINGSUCHINTERACTIONSHAVENOTBEENWELLCHARACTERIZEDWEREPORTHERETHATEPHRINA5,ALIGANDOFTHEEPHRECEPTORTYROSINEKINASES,PLAYSAKEYROLEINLENSFIBERCELLSHAPEANDCELL–CELLINTERACTIONSLENSFIBERCELLSINMICELACKINGEPHRINA5FUNCTIONAPPEARROUNDEDANDIRREGULARINCROSSSECTION,INCONTRASTTOTHEIRNORMALHEXAGONALAPPEARANCEINWTLENSESCATARACTSEVENTUALLYDEVELOPIN87OFEPHRINA5KOMICEWEFURTHERDEMONSTRATETHATEPHRINA5INTERACTSWITHTHEEPHA2RECEPTORTOREGULATETHEADHERENSJUNCTIONCOMPLEXBYENHANCINGRECRUITMENTOF?CATENINTONCADHERINTHESERESULTSINDICATETHATTHEEPHRECEPTORSANDTHEIRLIGANDSARECRITICALREGULATORSOFLENSDEVELOPMENTANDMAINTENANCE?CATENIN?EPHRECEPTOR?NCADHERINCATARACT,ORTHEOPACIFICATIONOFTHELENS,ISTHELEADINGCAUSEOFVISUALIMPAIRMENTANDBLINDNESSWORLDWIDE1THEMOLECULAREVENTSUNDERLYINGLENSDEVELOPMENTANDTHEPROCESSESBYWHICHTHELENSMAINTAINSTRANSPARENCYOVERALIFETIMEAREUNCLEAR2INADDITION,THECELLULARANDBIOCHEMICALMECHANISMSUNDERLYINGTHEPATHOLOGICALCHANGESLEADINGTOCATARACTREMAINPOORLYUNDERSTOODTHELENSISCOMPOSEDOFASINGLELAYEROFEPITHELIALCELLSONTHEANTERIORSURFACE,WHICH,OVERALIFETIME,DIVIDEANDDIFFERENTIATEINTOTHEUNDERLYINGLENSFIBERCELLSTHATCOMPRISETHEBULKOFTHELENS3,4INITIALLYDURINGLENSDEVELOPMENT,PRIMARYLENSFIBERCELLSDIFFERENTIATEANDELONGATEFROMTHEPOSTERIORPOLEINLATEREMBRYOGENESISANDTHROUGHOUTLIFE,SECONDARYLENSFIBERCELLSDIFFERENTIATEFROMLENSEPITHELIALCELLSLOCATEDATTHEEQUATORINCROSSSECTION,THELENSFIBERCELLSRESEMBLEFLATTENEDHEXAGONSWITHTWOBROADANDFOURSHORTSIDES3THESECELLSAREORGANIZEDINAHIGHLYORDEREDANDCLOSELYPACKEDMANNER,ANDINTERACTTHROUGHEXTENSIVEINTERCELLULARADHESIONCOMPLEXESINCLUDINGGAPANDADHERENSJUNCTIONS5FIBERCELLGAPJUNCTIONSARECOMPOSEDOFCONNEXINSCX46AND506,INACTIVATIONOFWHICHLEADSTOTHEDEGENERATIONOFTHEINNERFIBERCELLSANDTHEDEVELOPMENTOFCATARACTINMICE7,8MUTATIONSINHUMANCXGENESHAVEALSOBEENASSOCIATEDWITHCATARACTOGENESIS9,10ASTHELENSISCOMPLETELYENCLOSEDBYANACELLULAR,AVASCULARCAPSULE,ITISBELIEVEDTHATTHESECELL–CELLJUNCTIONSARECRITICALFORPROVIDINGNUTRIENTTRANSPORT,REMOVALOFMETABOLICWASTES,ANDMAINTENANCEOFHOMEOSTASIS11,12INADDITIONTOGAPJUNCTIONS,WIDESPREADADHERENSJUNCTIONSCONTAININGNCADHERINANDITSASSOCIATEDPROTEIN?CATENINEXISTBETWEENLENSFIBERCELLS13–16,ANDMAYPLAYIMPORTANTROLESINLENSDEVELOPMENTANDFUNCTIONALTHOUGHCELL–CELLINTERACTIONISCRITICALFORMAINTAININGLENSTRANSPARENCY,LITTLEISKNOWNABOUTTHEMOLECULARMECHANISMSUNDERLYINGTHESEINTERACTIONSWEHAVEIDENTIFIEDANUNEXPECTEDREGULATOROFLENSFIBERCELL–CELLINTERACTION,THEAXONGUIDANCEMOLECULEEPHRINA517–19,ANDHAVESHOWNTHATTHELOSSOFITSFUNCTIONLEADSTOALTERATIONSOFCELLSHAPEANDSEVERECATARACTSINTHEADULTMOUSEOURSTUDIESIDENTIFYANOVELFUNCTIONOFEPHRINA5INLENSDEVELOPMENTANDSUGGESTUNIQUEREGULATIONOFDOWNSTREAMSIGNALINGMECHANISMSWESHOWHERETHATADISRUPTIONINEPHA2–EPHRINA5INTERACTIONLEADSTOTHEINTERNALIZATIONOFNCADHERINANDADISRUPTIONINTHEBINDINGOFNCADHERINWITH?CATENINRESULTSANDDISCUSSIONEPHRINA5?/?MICEDEVELOPCATARACTSEXAMINATIONOFEPHRINA5?/?MUTANTMICEUSINGSLITLAMPBIOMICROSCOPYANDSCHEIMPFLUGIMAGINGREVEALEDLARGEREGIONSOFOPACIFICATIONINTHEADULTMUTANTLENSESFIG1A–DSUCHCATARACTSDEVELOPEDIN87OFMUTANTMICEOLDERTHAN6MONTHSN?22,BUTNOTINANYWTCONTROLSORHETEROZYGOUSANIMALSN?24THEOVERALLSIZEANDMORPHOLOGYOFTHEHETEROZYGOUSLENSESWEREINDISTINGUISHABLEFROMTHATOFTHEWTLENSINTHEMUTANTLENS,HISTOLOGICALANALYSISREVEALEDRUPTURESOFTHEPOSTERIORLENSCAPSULEANDLENSDISRUPTIONSWITHVARYINGDEGREESOFSEVERITYINTHEMUTANTMICEFIG1F,G,I,ANDJINTHEMOSTSEVERECASES,THELENSCOMPLETELYDEGENERATED,LEAVINGTISSUEREMNANTSIMPINGINGAGAINSTTHERETINAANDSOMETIMESTHEIRISLOSSOFCELLSHAPECONTROLINEPHRINA5?/?LENSESTOEXAMINETHENATUREANDTIMINGOFTHEINITIALDEFECTS,LENSESFROMWTANDEPHRINA5?/?MICEWERECOLLECTEDATVARIOUSDEVELOPMENTALSTAGESE14,E17,P0,P6,P21,P30,ANDP60,SECTIONED,ANDSTAINEDWITHHMAC,AIS,YS,NJK,ANDRZPERFORMEDRESEARCHDKANDNJKCONTRIBUTEDNEWREAGENTS/ANALYTICTOOLSMAC,AIS,NJK,ANDRZANALYZEDDATAANDMAC,AIS,NJK,ANDRZWROTETHEPAPERTHEAUTHORSDECLARENOCONFLICTOFINTEREST1MACANDAISCONTRIBUTEDEQUALLYTOTHISWORK2TOWHOMCORRESPONDENCESHOULDBEADDRESSEDEMAILRZHOURCIRUTGERSEDUTHISARTICLECONTAINSSUPPORTINGINFORMATIONONLINEATWWWPNASORG/CGI/CONTENT/FULL/0808987105/DCSUPPLEMENTAL?2008BYTHENATIONALACADEMYOFSCIENCESOFTHEUSA16620–16625?PNAS?OCTOBER28,2008?VOL105?NO43WWWPNASORG?CGI?DOI?101073?PNAS0808987105ECADHERINCANALSOBEINTERNALIZED30–32ADDITIONALLY,NMDARECEPTORACTIVITYINCREASEDNCADHERINTURNOVERTHROUGHENDOCYTOSISTOMODULATEADHESION33OUROBSERVATIONSHERESUGGESTTHATEPHRINA5FUNCTIONSTOPROMOTENCADHERINMEMBRANELOCALIZATIONDURINGLENSDEVELOPMENTDECREASEDEPHA2ACTIVATIONINEPHRINA5?/?LENSESTOIDENTIFYWHICHEPHRECEPTORSMEDIATEEPHRINA5FUNCTIONINLENSDEVELOPMENT,WEEXAMINEDTHEEXPRESSIONOFEPHRECEPTORSINWTLENSESBYPCREXPRESSIONOFEPHA2,EPHA3,EPHA5,EPHA7,EPHA8,ANDALLEPHBRECEPTORSWASDETECTEDNOTSHOWNEXAMINATIONOFLENSESFROMEPHA3ABROWN,PERSONALCOMMUNICATION,EPHA5,ANDEPHB1NULLMICEFAILEDTODETECTANYMORPHOLOGICALDEFECTSTHEREFORE,WEPROCEEDEDTOEXAMINETHEEXPRESSIONOFEPHA2INTHEDEVELOPINGLENSTODETERMINEWHEREEPHA2PROTEINWASEXPRESSED,WEPERFORMEDDOUBLEIMMUNOFLUORESCENCESTUDIESFORSUBCELLULARLOCALIZATIONOFBOTHEPHA2RECEPTORANDEPHRINA5PROTEINSINTHEP21LENSEPHA2PROTEINWASDETECTEDWITHAGOATANTIEPHA2ANTIBODYCOUPLEDWITHACY3CONJUGATEDANTIGOATSECONDARYANTIBODYFORANALYSISOFABBCCDDE‘‘‘FIG4BOTHEPHA2ANDEPHRINALIGANDSAREEXPRESSEDATTHECELLJUNCTIONSAPHALLOIDINSTAININGOFWTLENSSHOWSLENSFIBERCELLORGANIZATIONBANDCWTTRANSVERSESECTIONSOFP21LENSESSTAINEDWITHANTIEPHA2ANDEPHA3FC,RESPECTIVELYLOWMAGNIFICATIONIMAGESDEMONSTRATETHATBOTHEPHA2ANDAEPHRINSARENORMALLYEXPRESSEDATHIGHERLEVELSINTHESUBCORTICALREGIONDEPHA3FCSTAININGONEPHRINA5?/?LENSSECTIONSSTAININGWASMOSTLYLOSTONMUTANTLENSESINDICATINGTHATTHESUBCORTICALSIGNALSWEREARESULTOFEPHRINA5EXPRESSIONB?–D?HIGHMAGNIFICATIONCONFOCALIMAGESOFB–DNOTETHATWTEPHA2RECEPTORB?ANDEPHRINA5C?EXPRESSIONISTHEHIGHESTATTHECELL–CELLJUNCTIONSEWTCONTROLWITHOUTPRIMARYANTIBODYIMAGESWERECOLLECTEDWITHEQUALEXPOSURETIMESARROWSINADENOTETHESUBCORTICALSCLENSFIBERREGIONFORA–ESCALEBARINTOPLEFT,20?MTOPRIGHT,5?MABFIG3CHANGEINNCADHERINLOCALIZATIONINEPHRINA5?/?LENSAALTEREDPATTERNSOFEXPRESSIONOFNCADHERINANDTHEGAPJUNCTIONPROTEINZO1INEPHRINA5?/?LENSESP21WTANDEPHRINA5?/?LENSCRYOSECTIONSWEREPREPARED10?MTHICKANDSTAINEDWITHANTI–NCADHERINANDANTI–ZO1ANTIBODIESBFRACTIONSOFNCADHERINSIGNALSDETECTEDINTHECYTOPLASMINP21WTANDEPHRINA5?/?LENSESTHEFRACTIONSWEREOBTAINEDBYDIVIDINGTHEFLUORESCENTSIGNALSINTHECYTOPLASMBYTHESIGNALSOFTHEENTIRECELLCELLBOUNDARIESAREDEFINEDBYSTAININGWITHALEXAFLUOR546PHALLOIDINSIGNIFICANTATP?005TTESTSCALEBARINA,5?M16622?WWWPNASORG?CGI?DOI?101073?PNAS0808987105COOPERETAL

簡介：中文中文7700字出處出處NEPHROLOGY152010599–608MICRORNA在腎臟疾病中的作用在腎臟疾病中的作用JORDANYZLI,TUCKYYONG,MICHAELZMICHAELANDJONATHANMGLEADLE摘要MICRORNA是一類非編碼小RNA家族，具有通過抑制靶基因的表達(dá)封鎖蛋白質(zhì)的翻譯或誘導(dǎo)MRNA的降解來調(diào)節(jié)生理和病理的過程。這些MIRNAS有調(diào)節(jié)成千上萬的蛋白質(zhì)表達(dá)的能力。因此，MIRNA能迅速成為與腎臟疾病相關(guān)的一個(gè)新的生物醫(yī)學(xué)領(lǐng)域。MIRNA的表達(dá)已經(jīng)顯示腎臟與其他器官間的不同及腎臟不同部分間的不同。此外，發(fā)現(xiàn)MIRNA在足細(xì)胞病變發(fā)展、糖尿病腎病及多囊腎的模型中都具有重要作用。特別是在小鼠的模型實(shí)驗(yàn)中足細(xì)胞中的一種MIRNA生物合成的關(guān)鍵酶DICER酶的特異性刪除，會導(dǎo)致蛋白尿及一些嚴(yán)重的腎功能障礙。MIRNA192可以作為轉(zhuǎn)化生長因子Β在糖尿病腎病高糖環(huán)境下活化的效應(yīng)器。據(jù)報(bào)道在腎臟移植排斥反應(yīng)中有不同的MIRNA的表達(dá)。估計(jì)未來涉及MIRNA的研究將會是研究各種腎臟疾病及各種診斷標(biāo)志物、治療靶點(diǎn)的新切點(diǎn)。本文就MIRNA在腎臟疾病中的發(fā)展及對診斷可能的影響及未來腎臟疾病的治療進(jìn)行綜述。緒論緒論MIRNA是內(nèi)源性非編碼RNA分子，長度為2022核苷酸。在過去的十年對MIRNA的發(fā)現(xiàn)及研究對我們理解基因的調(diào)節(jié)、細(xì)胞增殖、分化、凋亡、代謝、許多包括腎臟疾病的病理生理都是革命性的影響。MIRNA的生物效應(yīng)及其在各種疾病中的作用的研究仍處于初步階段，但是發(fā)展得非常迅速。本文目的是，介紹MIRNA與腎臟疾病相關(guān)的生物學(xué)作用理解其在疾病發(fā)生中的機(jī)制，及未來這一領(lǐng)域討論的方向。MIRNA的發(fā)現(xiàn)和生物合成的發(fā)現(xiàn)和生物合成1993年MIRNA首次發(fā)現(xiàn)于CAENORHABDITISELEGANS線蟲中。從那時(shí)起MIRNA也廣泛發(fā)現(xiàn)于植物和哺乳動(dòng)物中。MIRNA首先轉(zhuǎn)錄成是含有莖環(huán)結(jié)構(gòu)的MIRNA前體，經(jīng)過DICER酶核糖核酸酶裂解成更短的前提RNA，這種酶重要的輔助因子叫做DGCR8（DIGEORGESYNDROMECRITICALREGION8），一種雙鏈RNA結(jié)合蛋白（圖一）。前體MIRNA通過輸出蛋白5轉(zhuǎn)運(yùn)出細(xì)胞核，在細(xì)胞質(zhì)中再通過DICER酶，另一種酶RNASEIII，切割為2022核苷酸長度的成熟形式。切割后，MIRNA的雙鏈退繞，功能連加載到RNA誘導(dǎo)的沉默復(fù)合體（RISC）。成熟的MIRNA引導(dǎo)RISC復(fù)合體到互補(bǔ)序列，通常是靶MRNA3‘末端翻譯區(qū)。結(jié)合后，RISC復(fù)合體導(dǎo)致轉(zhuǎn)錄后基因沉默通過切割靶MRNA或者一直其翻譯，因此MIRNA通常是負(fù)調(diào)控基因。除了在轉(zhuǎn)錄后表達(dá)中的作用，MIRNA已經(jīng)應(yīng)用于轉(zhuǎn)錄基因沉默通過針對啟動(dòng)區(qū)域，而且據(jù)報(bào)道對轉(zhuǎn)錄有正調(diào)控作用。每一個(gè)MIRNA都有調(diào)控大量不同MRNA翻譯的潛力，每個(gè)MRNA又擁有大量的對不同MRNA的單結(jié)合位點(diǎn)，因?yàn)镸IRNA的特異性主要由在5‘末端的WSTSONCRICK堿基配對決定。據(jù)估計(jì)，在人體內(nèi)的不同的MIRNA的序列超過1000個(gè)。經(jīng)過分析預(yù)測，超過60的人累計(jì)因都是MIRNAS的潛在靶點(diǎn)，并且有大量的其他比MIRNA長的非編碼RNA，它們也具有重要的功能。然而，直接的實(shí)驗(yàn)證據(jù)表明，MIRNA調(diào)控的MRNA只是一小部分的MIRNA和目標(biāo)MRNA。檢測檢測MIRNA的水平的水平最初分析特定的MIRNA序列的水平是十分繁瑣的，但是現(xiàn)在隨著技術(shù)的進(jìn)步，現(xiàn)在在檢測的敏感性及特異性都改善到可以在臨床上應(yīng)用。最初，RNA印記雜交提供了在總RNA樣本中定量及定性的關(guān)于大量MIRNA各種形式的分析。由于MIRNA在MIRBASE注冊表中的增加，芯片技術(shù)已經(jīng)可以平行檢測成千上萬的MIRNA在一個(gè)樣本中。最近，實(shí)時(shí)逆轉(zhuǎn)錄聚合酶鏈反應(yīng)已經(jīng)被應(yīng)用于實(shí)時(shí)定量和定性分析MIRNA的水平。成熟的MIRNA特異性擴(kuò)增可以利用莖環(huán)結(jié)構(gòu)的逆轉(zhuǎn)錄及熒光定量檢測實(shí)現(xiàn)，而代替試劑用于初級轉(zhuǎn)錄產(chǎn)張力蛋白（PTEN），這是MIR216A和MIR217的靶點(diǎn)。反過來，這些MIRNA通過TGFΒ上調(diào)，間接通過MIR192，在小鼠系膜細(xì)胞中。在其他的動(dòng)物研究中通過體內(nèi)外實(shí)驗(yàn)，張等人已經(jīng)證實(shí)在糖尿病腎病的早期MIR21的表達(dá)會下調(diào)。MIR21的過表達(dá)抑制高糖環(huán)境下腎小球系膜細(xì)胞的增殖。糖尿病DB/DB小鼠24小時(shí)尿白蛋白排泄率降低了在提高了MIR21的暴露后。相同的研究還發(fā)現(xiàn)PTEN為MIR021的目標(biāo)基因。另一項(xiàng)研究已經(jīng)證明在人類和小鼠腎小球系膜細(xì)胞中高的葡萄糖水平科引起MIR377過表達(dá)。MIR377已經(jīng)證明減少了P21活化激酶（PAK1）和錳超氧化物歧化酶的表達(dá)。這將增強(qiáng)纖維連接蛋白產(chǎn)生，它是糖尿病腎病系膜細(xì)胞的特點(diǎn)。我們估計(jì)在足細(xì)胞、腎小管、其他腎細(xì)胞中許多其他的MIRNA表達(dá)在高血糖的條件下將解除調(diào)節(jié)。在糖尿病腎病中，改變MIRNA的表達(dá)對一些病理生理狀態(tài)的反應(yīng)是令人感興趣的，特別是缺氧缺血和高糖的刺激。王和他的同事們的發(fā)現(xiàn)第一次看到了高糖對MIRNA在系膜細(xì)胞中的表達(dá)。此外，已經(jīng)證明高血糖通過MIR221影響內(nèi)皮功能障礙。多囊腎多囊腎常染色體顯性多囊腎?。ˋDPKD）是最常見的遺傳腎臟疾病之一。通常，在多囊腎1基因的突變（PKD1）占ADPKD的85，而在多囊腎2基因（PKD2）突變?yōu)槭Ｓ嗟牟糠帧KD2編碼的蛋白質(zhì)稱為多囊蛋白2。多囊蛋白2的異常表達(dá)引起腎小管和膽管上皮的異常增生，通常會導(dǎo)致囊腫形成。最近發(fā)現(xiàn)MIRNA在控制PKD基因的表達(dá)及調(diào)節(jié)功能作用中有潛在的作用。兩個(gè)小組已經(jīng)證明MIR17直接的靶點(diǎn)PKD2的3‘UTR轉(zhuǎn)錄后抑制PKD2的表達(dá)。此外，他們還證明了MIR17的過表達(dá)可能促進(jìn)細(xì)胞增殖通過轉(zhuǎn)錄后抑制PKD2在HEK293T細(xì)胞（人胚腎細(xì)胞）中。尋找新的以PKD1為靶點(diǎn)的MIRNA已經(jīng)成為一個(gè)熱點(diǎn)研究領(lǐng)域。用個(gè)PKD模型的大鼠，相對于健康的大鼠在腎臟組織中30個(gè)差異表達(dá)的MIRNA已經(jīng)被證實(shí)，其中29個(gè)都下調(diào)。兩種算法TARGETSCAN和MIRANDA，預(yù)測目標(biāo)為在PKD中不受調(diào)節(jié)的MIRNA與通路影響有關(guān)的用KEGG、GO、BIOCARTA和分子簽名數(shù)據(jù)庫。在PKD中不被調(diào)節(jié)的MIRNA與基因的24個(gè)功能類別都有關(guān)系，包括一些通路，對囊腫形成重要的如MTOR信號、絲裂原活化蛋白激酶信號、WNT信號和TGFΒ通路。但是這些相關(guān)性都需要實(shí)驗(yàn)驗(yàn)證。MIR15A已經(jīng)報(bào)道調(diào)節(jié)細(xì)胞周期調(diào)控的CDC25A和影響肝囊腫生成在大鼠PKD模型中。在原位雜交中表明MIR15A下調(diào)在ADPKD、常染色體隱性遺傳多囊腎、者肝纖維化同時(shí)患者PKD的患者的肝臟組織中。相反地，MIR15A在細(xì)胞中的過表達(dá)從PKD大鼠中派生導(dǎo)致了CDC25A蛋白的下降，小幅下降在G1S期轉(zhuǎn)變和細(xì)胞增殖期，較大的下降在體外的的生長的囊腫中。這種在囊腫中不成比例的生長表明下降的MIR15A或許促進(jìn)囊腫形成的增加通過除了細(xì)胞增殖的其他機(jī)制。其他腎臟疾病其他腎臟疾病試圖理解MIRNA在腎臟疾病中的作用，一種顯而易見的方法可以比較在來自正常和受影響的患者的樣本之間的MIRNA表達(dá)。在腎臟疾病中，這個(gè)研究包括了患有IGA腎臟、狼瘡性腎炎、高血壓和腎癌的病人。戴和他的同事進(jìn)行了一項(xiàng)研究比較了11個(gè)IGA腎病患者活檢標(biāo)本與3個(gè)控制組的MIRNA表達(dá)。他們能夠證明在132個(gè)IGA腎病和正常對照組腎臟組織樣本中，其中31個(gè)MIRNA下調(diào)，35個(gè)上調(diào)。最近，另一項(xiàng)研究報(bào)告了MIR200C，MIR141，MIR205和MIR192不同的腎內(nèi)的表達(dá)，發(fā)現(xiàn)這些和腎臟疾病的嚴(yán)重性及進(jìn)展相關(guān)。MIR200C和MIR205的去調(diào)控表達(dá)另我們感興趣的是上皮向間質(zhì)轉(zhuǎn)化的鏈接。66MIRNA已經(jīng)被發(fā)現(xiàn)差異表達(dá)在小量的來自人II類狼瘡性腎炎患者的腎臟組織與健康對照組相比。在外周血的單核細(xì)胞中MIRNA（16MIRNA，7個(gè)下調(diào)，9個(gè)上調(diào)）的差異

簡介：中文中文2830字出處出處WANGJ,GUODONGLIUAULTRASENSITIVEELECTRICALBIOSENSINGOFPROTEINSANDDNA鈥CARBONNANOTUBEDERIVEDAMPLIFICATIONOFTHERECOGNITIONANDTRANSDUCTIONEVENTSJJOURNALOFTHEAMERICANCHEMICALSOCIETY,2004,1261030101超靈敏免疫和超靈敏免疫和DNADNA電化學(xué)生物分析電化學(xué)生物分析應(yīng)用碳納米管放大分子識別和傳導(dǎo)過程應(yīng)用碳納米管放大分子識別和傳導(dǎo)過程蛋白質(zhì)和DNA檢測技術(shù)在基因疾病的診斷和治療，傳染性疾病的檢測，藥物的開發(fā)，生物戰(zhàn)爭預(yù)警中起著非常重要的作用。這些生物檢測通常依賴DNA雜交或抗原抗體相互作用，可達(dá)到超靈敏度的檢測。由于電化學(xué)傳感器具有靈敏度高、簡單、可微型化，低成本和高需求的特點(diǎn)，非常適用于生物檢測。酶標(biāo)記在蛋白質(zhì)和DNA超靈敏電化學(xué)生物親和性檢測中起著很大的作用。HLELLER，小組5，6通過DNA上連接HRP酶標(biāo)記物和采用一種可加快電子傳遞的氧化聚合物可實(shí)現(xiàn)DNA的高靈敏度的電化學(xué)檢測（低至5ZMOL）。WILLNER,S小組7～8通過生物催化沉積酶反應(yīng)產(chǎn)物可獲得信號的多重放大，從而實(shí)現(xiàn)極低的檢測限（25AMOL），酶聯(lián)電化學(xué)蛋白質(zhì)檢測信號可通過雙酶底物體系底物循環(huán)或者酶產(chǎn)物的離子交換富集產(chǎn)物來進(jìn)行放大。然而，在電化學(xué)生物檢測中，放大生物識別傳導(dǎo)信號仍是一個(gè)重大的挑戰(zhàn)。為了滿足蛋白質(zhì)和核酸電化學(xué)檢測高靈敏度的要求，我們需要新的方法，通過酶生物催化反應(yīng)來放大信號。在本文中，我們利用碳納米管（CNTS）顯著放大蛋白質(zhì)和DNA的識別和電化學(xué)傳導(dǎo)信號。CNTS所特有的電學(xué)性能、化學(xué)性能和機(jī)械性質(zhì)使其非常適用于電化學(xué)傳感器1，2。大多數(shù)CNT傳感器主要是利用CNTS特有的表面性質(zhì)來促進(jìn)在生物催化裝置中電子轉(zhuǎn)移反應(yīng)，在我們新的生物親和性檢測中（圖1），CNTS起著放大識別和傳導(dǎo)信號的雙重作用，也就是CNTS上載有大量酶及積累了大量酶反應(yīng)的產(chǎn)物。這些新奇的方法和CNTS預(yù)富集的功能反映了CNTS具有很大的比表面積，并可用ALP酶標(biāo)來證實(shí)。通過采用CNTS的放大處理從而降低檢測限的一些方法已被報(bào)道過，因此很適用于電化學(xué)DNA檢測。圖，這些微觀圖片是HITACHIH7000儀器在工作電壓為75KV下拍下的。從圖3DNA分子雜交（A）和抗原抗體生物檢測中可以看到由于CNT的雙重放大作用引起了傳感信號的明顯增強(qiáng)?；趩蚊笜?biāo)記物和一個(gè)玻碳電極的傳統(tǒng)檢測方法既不能對10PGML1的目標(biāo)DNA（A，A）也不能對80PGML1的IGG（B，B）產(chǎn)生響應(yīng)?；谳d有ALP酶的CNTS（B）的第一放大步驟為這些分析物的低濃度檢測提供了方便。單ALP酶檢測即使在分析物濃度較高（1000倍）的條件下仍顯示一個(gè)較低的信號（圖中未標(biāo)出）。改進(jìn)方法后的檢測靈敏度達(dá)到將近104，正好與每個(gè)CNT上載有的ALP酶估計(jì)量相一致。用涂覆上鏈霉親合素的聚苯乙烯代替CNT來裝載粒子獲得的靈敏度增強(qiáng)約僅為單酶檢測的50倍。在信號第二放大途徑，即用CNT修飾傳感器可獲得更強(qiáng)的DNA和蛋白質(zhì)檢測信號（約是用鏈霉親合素修飾聚苯乙烯檢測的30倍）（C）。后者反映了在CNT層強(qiáng)烈地吸附著大量游離的Α奈酚。在CNT上富集產(chǎn)物的現(xiàn)象可用由沉積時(shí)間的不同而引起的Α奈酚信號的突增來描述（與裸電極上產(chǎn)生的時(shí)間信號關(guān)系相比較；見圖2的支持信息）。圖310PGML1目標(biāo)寡核苷酸（A）和80PGML1IGG（B）分別在玻碳電極（A）單ALP酶標(biāo)（B）和載有大量ALP酶標(biāo)的CNT上用計(jì)時(shí)電勢分析法進(jìn)行檢測產(chǎn)生的信號。在檢測中（C）除了使用CNT修飾的玻碳電極外與（B）均相同。磁性粒子量，50UG；分別進(jìn)行20和30分鐘的DNA雜交和抗原/抗體免疫反應(yīng)；樣品體積，50UL。檢測，往樣品中加入50ULΑ奈基磷酸鹽（50MM）溶液進(jìn)行酶反應(yīng)20分鐘。產(chǎn)物Α奈酚的測量在裸或者是已用

簡介：MICROSATELLITEINSTABILITYAND/ORLOSSOFHETEROZYGOSITYINYOUNGGASTRICCANCERPATIENTSINITALYYIHHORNGSHIAO1,DANIELABOVO2,MARIAGUIDO2,CARLOCAPELLA3,MAUROCASSARO2,GRAZIELLABUSATTO2,VALENTINARUSSO2,4,ANGELOSIDONI5,ANNARPARENTI2ANDMASSIMORUGGE21LABORATORYOFCOMPARATIVECARCINOGENESIS,NCIFCRDC,NATIONALINSTITUTESOFHEALTH,FREDERICK,MD,USA2DEPARTMENTOFONCOLOGICALANDSURGICALSCIENCES,CATTEDRADIISTOCHIMICAEIMMUNOHISTOCHIMICAPATOLOGICA,IIICATTEDRADIANATOMIAPATOLOGICA,UNIVERSITA′DEGLISTUDI,PADOVA,ITALY3DEPARTMENTOFCLINICALANDBIOLOGICALSCIENCE,UNIVERSITYOFPAVIA,VARESE,ITALY4DEPARTMENTOFPATHOLOGY,UNIVERSITYOFCATANIA,CATANIA,ITALY5DEPARTMENTOFPATHOLOGY,UNIVERSITYOFPERUGIA,PERUGIA,ITALYGASTRICCANCERSARERARELYDIAGNOSEDBEFORETHEAGEOF40YEARSANDTHEINCIDENCEREACHESAPEAKDURINGTHE7THDECADEINTHEGENERALPOPULATIONAMOLECULARMECHANISMOFEARLYTUMORONSETMAYBEDETERMINEDBYCOMPARINGMICROSATELLITEINSTABILITYMSI,INDICATIVEOFERRORPRONEMISMATCHREPAIR,ANDLOSSOFHETEROZYGOSITYLOHBETWEENGASTRICCANCERSINPATIENTSI40YEARSOFAGEANDTHOSEOFOLDERAGESTHREETO5CHROMOSOMALLOCI,WHEREMSIAND/ORLOHARECOMMONLYFOUNDINGASTRICCANCERSINTHEGENERALPOPULATION,WEREEXAMINEDINFORMALINFIXED,PARAFFINEMBEDDEDSAMPLESFROM102PATIENTSI40YEARSOFAGEUSINGAPOLYMERASECHAINREACTIONBASEDNONRADIOACTIVESCREENINGMETHODMSIAND/ORLOHATAMINIMUMOF1LOCUSWEREDETECTEDIN11/102PATIENTSTHEFREQUENCYOFMSIAND/ORLOHATTHED11S904LOCUSWASSIGNIFICANTLYHIGHERTHANTHATATTHED2S119,D2S123,D5S409ANDIFNAREGIONSNOPREFERENTIALGENETICCHANGESATTHED11S904LOCUSWEREOBSERVEDINELDERLYPATIENTSAMONGSEVERALCLINICOPATHOLOGICALVARIABLES,ASTATISTICALLYSIGNIFICANTASSOCIATIONWITHMSIAND/ORLOHWASOBSERVEDONLYFORTUMORSLOCATEDATTHECARDIA,COMPAREDWITHTUMORSATTHEANTRUMANDTHECORPUSOURFINDINGSSUGGESTTHATAUNIQUEMECHANISMMAYBEINVOLVEDININCREASINGTHESUSCEPTIBILITYOFTHED11S904LOCUSFOREITHERMSIORLOH,ESPECIALLYFORCARDIATUMORSINYOUNGPATIENTSEARLYONSETOFGASTRICCANCERSINPATIENTSI40YEARSOFAGEISASSOCIATEDWITHGENETICCHANGESATPREFERENTIALCHROMOSOMALLOCI,INCLUDINGD11S904INTJCANCER8259–62,1999?1999WILEYLISS,INCALTHOUGHGASTRICCANCERINCIDENCEHASBEENDECLINING,ITWASSTILLRANKEDASTHE2NDMOSTCOMMONCANCERANDTHE2NDLEADINGCAUSEOFCANCERDEATHINTHEWORLDINTHE1990SPARKINETAL,1993BECAUSEOFTHEAGGRESSIVENATUREOFGASTRICCANCERS,THEOVERALL5YEARSURVIVALRATEISLESSTHAN20BREAUXETAL,1990GASTRICCANCERSUSUALLYOCCURINPATIENTSOLDERTHAN50YEARSOFAGEONLYABOUT5OFGASTRICCANCERPATIENTSAREYOUNGERTHAN40YEARSNEUGUTETAL,1996GASTRICCANCERSINPATIENTS?40YEARSOFAGEAREMOREAGGRESSIVETHANTHOSEINELDERLYPATIENTSFUJIMOTOETAL,1994COMPARISONOFGENETICALTERATIONSINGASTRICCANCERSBETWEENPATIENTS?40YEARSOFAGEANDOLDERPATIENTSMAYIDENTIFYASPECIFICMOLECULARMECHANISMASSOCIATEDWITHANEARLYONSETOFTUMORMICROSATELLITESARESHORTTANDEMLYREPEATEDDNASEQUENCESANDPRESENTTHROUGHOUTMAMMALIANGENOMESREPETITIVESEQUENCESOFDI,TRIANDTETRANUCLEOTIDEREPEATSAREPRESENTINMORETHAN50OFTHEHUMANGENOMEBECKMANANDWEBER,1992THETANDEMREPEATSOFDCDANAREPARTICULARLYABUNDANTALTERATIONSINTHENUMBEROFREPEATSPERSITE,KNOWNASMICROSATELLITEINSTABILITYMSI,HAVEBEENIMPLICATEDINVARIOUSHUMANDISEASES,INCLUDINGNEOPLASMSSPEICHER,1995MICROSATELLITESEQUENCESAREALSOHIGHLYPOLYMORPHICPOLYMORPHISMDUETODIFFERENTNUMBEROFREPEATSISVERYUSEFULTODETERMINEGENETICALTERATIONS,SUCHASLOSSOFHETEROZYGOSITYLOHINTHISSTUDY,WEEXAMINED5CHROMOSOMALLOCI,WHEREMSIAND/ORLOHARECOMMONLYFOUNDINGASTRICCANCERSINTHEGENERALPOPULATIONRHYUETAL,1994CHONGETAL,1994NAGELETAL,1995TAMURAETAL,1995LINETAL,1995SERUCAETAL,1995BUONSANTIETAL,1997,INGASTRICCANCERPATIENTS?40YEARSOFAGETHEFREQUENCYOFMSIAND/ORLOHANDTHEIRASSOCIATIONWITHOTHERCLINICOPATHOLOGICALDATAAREDISCUSSEDMATERIALANDMETHODSPATIENTSFORMALINFIXED,PARAFFINEMBEDDEDTISSUEBLOCKSFROM102PATIENTS?40YEARSOFAGEMEANAGE35YEARSRANGE16–40YEARSWERERETRIEVEDFROMTHEARCHIVESOFMANYITALIANHOSPITALSWITHCOMPARABLEGASTRICCANCERINCIDENCEDEMOGRAPHICANDPATHOLOGICALDATA,INCLUDINGAGE,GENDER,SITEOFTUMORANDTUMORSTAGEUSINGTHETNMSYSTEMBEAHRSANDMYERS,1983,WEREOBTAINEDFROMACOLLABORATIVEMULTICENTERSTUDYTHESITEOFTUMORWASCATEGORIZEDINTOTHEANTRUM,CORPUSORCARDIAOFSTOMACHNOCANCEREXTENDEDBEYONDTHEGASTRICESOPHAGEALJUNCTIONACCORDINGTOTHELAURE′NSYSTEM1965,GASTRICCANCERSWERECLASSIFIEDASEITHERINTESTINALORDIFFUSETYPEWHENBOTHPHENOTYPESCOEXISTED,CLASSIFICATIONOFTHETUMORTYPEWASBASEDONTHEMOSTREPRESENTATIVEHISTOLOGYGASTRITISNONATROPHICVSATROPHIC/METAPLASTICWASCLASSIFIEDUSINGTHEHOUSTONUPDATEDSYDNEYSYSTEMDIXONETAL,1996ALLCASESWEREJOINTLYASSESSEDBY2AUTHORSMCANDMRDNAEXTRACTIONNEOPLASTICANDADJACENTNONNEOPLASTICAREASWEREMICRODISSECTEDSEPARATELYFROMUNSTAINEDFORMALINFIXED,PARAFFINEMBEDDEDSECTIONSFORDEPARAFFINIZATION,PROTEINASEKDIGESTIONANDDNAPURIFICATION,ASDESCRIBEDPREVIOUSLYSHIAOETAL,1994MICRODISSECTEDNEOPLASTICAREASCONTAINEDATLEAST50TUMORCELLSWHENPOSSIBLE,LYMPHOCYTESWEREUSEDASANONNEOPLASTICCELLCONTROLPOLYMERASECHAINREACTIONPCRFIVECHROMOSOMALLOCIWITHCANDINUCLEOTIDEREPEATSD2S119,D2S123,D5S409,IFNAANDD11S904WERESELECTEDFORPCRPRIMERSD2S123AFM093XH3AANDAFM093XH3MD5S409AFM184YB6AANDAFM184YB6MIFNAIFNAPCR21ANDIFNAPCR22D11S904AFM081ZA5AANDAFM081ZA5MWEREOBTAINEDFROMTHEGENOMEDATABASEWEBSITEWWWGDBORGFORD2S119,UPPER5?CCAGTTTGGAAGCATTT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