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,NMDARECEPTORACTIVITYINCREASEDNCADHERINTURNOVERTHROUGHENDOCYTOSISTOMOD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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封鎖蛋白質的翻譯或誘導MRNA的降解來調節(jié)生理和病理的過程。這些MIRNAS有調節(jié)成千上萬的蛋白質表達的能力。因此，MIRNA能迅速成為與腎臟疾病相關的一個新的生物醫(yī)學領域。MIRNA的表達已經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驗中足細胞中的一種MIRNA生物合成的關鍵酶DICER酶的特異性刪除，會導致蛋白尿及一些嚴重的腎功能障礙。MIRNA192可以作為轉化生長因子Β在糖尿病腎病高糖環(huán)境下活化的效應器。據(jù)報道在腎臟移植排斥反應中有不同的MIRNA的表達。估計未來涉及MIRNA的研究將會是研究各種腎臟疾病及各種診斷標志物、治療靶點的新切點。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行綜述。緒論緒論MIRNA是內源性非編碼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節(jié)、細胞增殖、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與腎臟疾病相關的生物學作用理解其在疾病發(fā)生中的機制，及未來這一領域討論的方向。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。從那時起MIRNA也廣泛發(fā)現(xiàn)于植物和哺乳動物中。MIRNA首先轉錄成是含有莖環(huán)結構的MIRNA前體，經過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結合蛋白（圖一）。前體MIRNA通過輸出蛋白5轉運出細胞核，在細胞質中再通過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誘導的沉默復合體（RISC）。成熟的MIRNA引導RISC復合體到互補序列，通常是靶MRNA3‘末端翻譯區(qū)。結合后，RISC復合體導致轉錄后基因沉默通過切割靶MRNA或者一直其翻譯，因此MIRNA通常是負調控基因。除了在轉錄后表達中的作用，MIRNA已經應用于轉錄基因沉默通過針對啟動區(qū)域，而且據(jù)報道對轉錄有正調控作用。每一個MIRNA都有調控大量不同MRNA翻譯的潛力，每個MRNA又擁有大量的對不同MRNA的單結合位點，因為MIRNA的特異性主要由在5‘末端的WSTSONCRICK堿基配對決定。據(jù)估計，在人體內的不同的MIRNA的序列超過1000個。經過分析預測，超過60的人累計因都是MIRNAS的潛在靶點，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，直接的實驗證據(jù)表明，MIRNA調控的MRNA只是一小部分的MIRNA和目標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)在隨著技術的進步，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用。最初，RNA印記雜交提供了在總RNA樣本中定量及定性的關于大量MIRNA各種形式的分析。由于MIRNA在MIRBASE注冊表中的增加，芯片技術已經可以平行檢測成千上萬的MIRNA在一個樣本中。最近，實時逆轉錄聚合酶鏈反應已經被應用于實時定量和定性分析MIRNA的水平。成熟的MIRNA特異性擴增可以利用莖環(huán)結構的逆轉錄及熒光定量檢測實現(xiàn)，而代替試劑用于初級轉錄產張力蛋白（PTEN），這是MIR216A和MIR217的靶點。反過來，這些MIRNA通過TGFΒ上調，間接通過MIR192，在小鼠系膜細胞中。在其他的動物研究中通過體內外實驗，張等人已經證實在糖尿病腎病的早期MIR21的表達會下調。MIR21的過表達抑制高糖環(huán)境下腎小球系膜細胞的增殖。糖尿病DB/DB小鼠24小時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的目標基因。另一項研究已經證明在人類和小鼠腎小球系膜細胞中高的葡萄糖水平科引起MIR377過表達。MIR377已經證明減少了P21活化激酶（PAK1）和錳超氧化物歧化酶的表達。這將增強纖維連接蛋白產生，它是糖尿病腎病系膜細胞的特點。我們估計在足細胞、腎小管、其他腎細胞中許多其他的MIRNA表達在高血糖的條件下將解除調節(jié)。在糖尿病腎病中，改變MIRNA的表達對一些病理生理狀態(tài)的反應是令人感興趣的，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在系膜細胞中的表達。此外，已經證明高血糖通過MIR221影響內皮功能障礙。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?。PKD2編碼的蛋白質稱為多囊蛋白2。多囊蛋白2的異常表達引起腎小管和膽管上皮的異常增生，通常會導致囊腫形成。最近發(fā)現(xiàn)MIRNA在控制PKD基因的表達及調節(jié)功能作用中有潛在的作用。兩個小組已經證明MIR17直接的靶點PKD2的3‘UTR轉錄后抑制PKD2的表達。此外，他們還證明了MIR17的過表達可能促進細胞增殖通過轉錄后抑制PKD2在HEK293T細胞（人胚腎細胞）中。尋找新的以PKD1為靶點的MIRNA已經成為一個熱點研究領域。用個PKD模型的大鼠，相對于健康的大鼠在腎臟組織中30個差異表達的MIRNA已經被證實，其中29個都下調。兩種算法TARGETSCAN和MIRANDA，預測目標為在PKD中不受調節(jié)的MIRNA與通路影響有關的用KEGG、GO、BIOCARTA和分子簽名數(shù)據(jù)庫。在PKD中不被調節(jié)的MIRNA與基因的24個功能類別都有關系，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。但是這些相關性都需要實驗驗證。MIR15A已經報道調節(jié)細胞周期調控的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下調在ADPKD、常染色體隱性遺傳多囊腎、者肝纖維化同時患者PKD的患者的肝臟組織中。相反地，MIR15A在細胞中的過表達從PKD大鼠中派生導致了CDC25A蛋白的下降，小幅下降在G1S期轉變和細胞增殖期，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或許促進囊腫形成的增加通過除了細胞增殖的其他機制。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。在腎臟疾病中，這個研究包括了患有IGA腎臟、狼瘡性腎炎、高血壓和腎癌的病人。戴和他的同事進行了一項研究比較了11個IGA腎病患者活檢標本與3個控制組的MIRNA表達。他們能夠證明在132個IGA腎病和正常對照組腎臟組織樣本中，其中31個MIRNA下調，35個上調。最近，另一項研究報告了MIR200C，MIR141，MIR205和MIR192不同的腎內的表達，發(fā)現(xiàn)這些和腎臟疾病的嚴重性及進展相關。MIR200C和MIR205的去調控表達另我們感興趣的是上皮向間質轉化的鏈接。66MIRNA已經被發(fā)現(xiàn)差異表達在小量的來自人II類狼瘡性腎炎患者的腎臟組織與健康對照組相比。在外周血的單核細胞中MIRNA（16MIRNA，7個下調，9個上調）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電化學生物分析電化學生物分析應用碳納米管放大分子識別和傳導過程應用碳納米管放大分子識別和傳導過程蛋白質和DNA檢測技術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，可達到超靈敏度的檢測。由于電化學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，非常適用于生物檢測。酶標記在蛋白質和DNA超靈敏電化學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酶標記物和采用一種可加快電子傳遞的氧化聚合物可實現(xiàn)DNA的高靈敏度的電化學檢測（低至5ZMOL）。WILLNER,S小組7～8通過生物催化沉積酶反應產物可獲得信號的多重放大，從而實現(xiàn)極低的檢測限（25AMOL），酶聯(lián)電化學蛋白質檢測信號可通過雙酶底物體系底物循環(huán)或者酶產物的離子交換富集產物來進行放大。然而，在電化學生物檢測中，放大生物識別傳導信號仍是一個重大的挑戰(zhàn)。為了滿足蛋白質和核酸電化學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）顯著放大蛋白質和DNA的識別和電化學傳導信號。CNTS所特有的電學性能、化學性能和機械性質使其非常適用于電化學傳感器1，2。大多數(shù)CNT傳感器主要是利用CNTS特有的表面性質來促進在生物催化裝置中電子轉移反應，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的產物。這些新奇的方法和CNTS預富集的功能反映了CNTS具有很大的比表面積，并可用ALP酶標來證實。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道過，因此很適用于電化學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?；趩蚊笜擞浳锖鸵粋€玻碳電極的傳統(tǒng)檢測方法既不能對10PGML1的目標DNA（A，A）也不能對80PGML1的IGG（B，B）產生響應。基于載有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較低的信號（圖中未標出）。改進方法后的檢測靈敏度達到將近104，正好與每個CNT上載有的ALP酶估計量相一致。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的DNA和蛋白質檢測信號（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層強烈地吸附著大量游離的Α奈酚。在CNT上富集產物的現(xiàn)象可用由沉積時間的不同而引起的Α奈酚信號的突增來描述（與裸電極上產生的時間信號關系相比較；見圖2的支持信息）。圖310PGML1目標寡核苷酸（A）和80PGML1IGG（B）分別在玻碳電極（A）單ALP酶標（B）和載有大量ALP酶標的CNT上用計時電勢分析法進行檢測產生的信號。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；分別進行20和30分鐘的DNA雜交和抗原/抗體免疫反應；樣品體積，50UL。檢測，往樣品中加入50ULΑ奈基磷酸鹽（50MM）溶液進行酶反應20分鐘。產物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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