簡介：ELECTROCHEMICALPROBEFORONCHIPTYPEFLOWIMMUNOASSAYIMMUNOGLOBULINGLABELEDWITHFERROCENECARBOALDEHYDEMINAOKOCHI,HIROKOOHTA,TSUYOSHITANAKA,TADASHIMATSUNAGADEPARTMENTOFBIOTECHNOLOGYANDLIFESCIENCES,TOKYOUNIVERSITYOFAGRICULTUREANDTECHNOLOGY,22416NAKACHO,KOGANEI,TOKYO1848588,JAPANTELEPHONE81423887020FAX81423857713EMAILTMATSUNACCTUATACJPRECEIVED2JUNE2004ACCEPTED13AUGUST2004PUBLISHEDONLINE25FEBRUARY2005INWILEYINTERSCIENCEWWWINTERSCIENCEWILEYCOMDOI101002/BIT20313ABSTRACTLABELINGOFFERROCENECARBOALDEHYDEFCCHOTOIMMUNOGLOBULINGIGGVIAFORMATIONOFSCHIFFBASEANDITSREDUCTIONWASINVESTIGATEDFORCONSTRUCTIONOFANELECTROCHEMICALPROBEFORMINIATURIZEDAMPEROMETRICFLOWIMMUNOASSAYAPPROXIMATELYEIGHTMOLECULESOFFCCHOWERELABELEDTOIGGANDTHEREVERSIBLEREDOXPROPERTYOFFERROCENEWASOBSERVEDLABELINGEFFICIENCYIMPROVEDBYOVERTHREETIMESASCOMPAREDTOTHECONVENTIONALMETHODUSINGFERROCENEMONOCARBOXYLICACIDFCCOOHALSO,BINDINGAFFINITYOFIGGLABELEDWITHFCCHOTOITSANTIGEN,IGE,WASINVESTIGATEDBYENZYMELINKEDIMMUNOSORBENTASSAYELISAANDSURFACEPLASMONRESONANCEASSAYIGGLABELEDWITHFCCHOTHATRETAINEDEIGHTFERROCENEMOIETYSHOWEDSUFFICIENTBINDINGAFFINITYTOITSANTIGENANDTHECURRENTRESPONSEOBTAINEDINTHEFLOWELECTROCHEMICALDETECTIONSYSTEMINCREASEDBY14FOLDASCOMPAREDWITHIGGLABELEDWITHFCCOOHWHENAPPLYINGTHEPOTENTIALOF390MVVSAG/AGCLTHEMINIMUMDETECTABLECONCENTRATIONOFIGGLABELEDWITHFCCHOWAS006AMIGGLABELEDWITHFCCHODEMONSTRATEBIOCHEMICALANDELECTROCHEMICALPROPERTIESTHATAREUSEFULFORELECTROCHEMICALIMMUNOSENSORSB2005WILEYPERIODICALS,INCKEYWORDSELECTROCHEMICALPROBESONCHIPIMMUNOASSAYFERROCENEINTRODUCTIONIMMUNOASSAYISONEOFTHEMOSTIMPORTANTMETHODSUSEDINCLINICALDIAGNOSES,ENVIRONMENTALANALYSES,ANDBIOCHEMICALSTUDIESIMMUNOSENSORSCANBECATEGORIZEDASOPTICAL,ELECTROCHEMICAL,ANDMICROGRAVIMETRICASSAY,BASEDONTHEDETECTIONPRINCIPLEAPPLIEDLUPPAETAL,2001CHEMILUMINESCENTORFLUORESCENTDETECTIONWITHENZYMELINKEDIMMUNOSORBENTASSAYELISAHASBEENTHEMOSTPRACTICALLYUSEDHOWEVER,ITREQUIRESPRECISEDETECTIONDEVICESFORMINIATURIZEDSYSTEMSWITHASMALLSAMPLEVOLUMETHEELECTROCHEMICALDETECTIONMETHODISSUITABLEFORSENSORMINIATURIZATIONANDAUTOMATEDDETECTION,SINCEITISHIGHLYSENSITIVE,LOWCOST,LOWPOWERREQUIREMENTS,ANDHASHIGHCOMPATIBILITYWITHADVANCEDMICROMACHININGTECHNOLOGIESDEVELOPMENTSINMINIATURIZATIONOFCHEMICALANDBIOTECHNOLOGICALPROCESSESHAVEASIGNIFICANTIMPACTONALLASPECTSOFDIAGNOSTICTESTINGMINIATURIZEDIMMUNOSENSORS,WHICHCOMBINETHEANALYTICALPOWEROFMICROFLUIDICDEVICESWITHTHEHIGHSPECIFICITYOFANTIBODYANTIGENINTERACTIONS,HAVEBEENINTENSIVELYDEVELOPEDBERNARDETAL,2001KOJIMAETAL,2003LIMETAL,2002,2003SALEHANDSOHN,2003SATOETAL,2002WANGETAL,1998,2002WANGANDJIN,2003FERROCENEDERIVATIVESHAVEOFTENBEENUSEDASELECTROCHEMICALSIGNALINGPROBESFORIMMUNOASSAYLIMETAL,2002,2003PADESTEETAL,2000WANGETAL,2002ASWELLASTHEDNAHYBRIDIZATIONASSAYFANETAL,2003KIMETAL,2003LONGETAL,2003TAKENAKAETAL,1994,2000,2003WANGETAL,2003LABELINGOFFERROCENEDERIVATIVESTOENZYMESSUCHASGLUCOSEOXIDASEHASBEENINTENSIVELYSTUDIEDANDUSEDASMEDIATORSINBIOSENSORSDEGANIANDHELLER,1987,1988,1989GLERIAETAL,1986SUZAWAETAL,1994ALSO,ELECTROACTIVELABELOFIGGWITHFERROCENEMONOCARBOXYLICACIDFCCOOHBYCHEMICALCROSSLINKERS,SULFONHYDROXYSULFOSUCCINIMIDENHSAND1ETHYL33DIMETHYLAMINOPROPYLCARBODIIMIDEHYDROCHLORIDEEDC,HASBEENCOMMONLYUSEDLIMETAL,2002,2003ONLYTWOTOTHREEFERROCENEMOIETYHASBEENSTABLYINTRODUCEDTOIGGANDITSBINDINGAFFINITYWASNOTWELLCHARACTERIZEDTHEREFORE,ANEWLABELINGMETHODISREQUIREDFORINTRODUCINGAHIGHERNUMBEROFFERROCENEMOIETYTOIGGFORSENSITIVEDETECTIONINTHEPRESENTSTUDY,IGGWASLABELEDWITHFERROCENECARBOALDEHYDEFCCHOFORSENSITIVEDETECTIONTHEELECTROCHEMICALPROPERTYOFIGGLABELEDWITHFCCHOWASINVESTIGATEDANDTHENUMBEROFBOUNDFERROCENEMOIETYONIGGWASESTIMATEDBYATOMICABSORPTIONSPECTROSCOPYALSO,BINDINGAFFINITYOFIGGLABELEDWITHFCCHOTOITSANTIGENWASCHARACTERIZEDUSINGFCCHO,ITWASPOSSIBLETOOBTAINAHIGHERELECTROCHEMICALSIGNALDUETOAHIGHERNUMBEROFLABELEDFERROCENEMOIETYONIGGB2005WILEYPERIODICALS,INCCORRESPONDENCETOTADASHIMATSUNAGACONTRACTGRANTSPONSORNEDOCONTRACTGRANTNUMBER30027JECTEDTOQUENCHTHEUNREACTEDSITESHBSEPBUFFER10MMHEPES,150MMNACL,34MEDTA,AND0005V/VTWEEN20WASUSEDASCONSTANTRUNNINGBUFFERFOLLOWINGBINDINGANDREGENERATIONTHESEQUENCEWASREPEATEDTOSEETHEINTERACTIONOFGOATANTIHUMANIGEIGGLABELEDWITHFCCHOANDITSANTIGEN,HUMANIGEIGGLABELEDWITHFCCHOANDNONLABELEDIGGATAPROTEINCONCENTRATIONINTHERANGEOF004–059AMWASINJECTEDFOR60ALATAFLOWRATEOF25AL/MININJECTIONOFANALYTEWASPERFORMEDFOR180SECFORMONITORINGASSOCIATIONCURVESANDDISSOCIATIONCURVESWEREMONITOREDFORANOTHER180SECBYINJECTIONOFHBSEPBUFFERREGENERATIONOFTHESENSORAFTERINJECTIONOFGOATANTIHUMANIGEIGGWASCONDUCTEDWITHA48SECPULSEOF10MMGLYCINEHCLBUFFERPH26REALTIMEREFERENCECURVESUBTRACTIONOVERANONCOATEDSURFACEWASEMPLOYEDELECTROCHEMICALDETECTIONOFIGGLABELEDWITHFCCHOANDFCCOOHCYCLICVOLTAMMETRYOFGOATANTIHUMANIGEIGGLABELEDWITHFERROCENEWASPERFORMEDIN100ALOFSAMPLEUSINGANELECTROCHEMICALANALYZERMODEL832A,BIOANALYTICALSYSTEMSBAS,WLAFAYETTE,INATASCANRATEOF100MV/SAGLASSYCARBONELECTRODEWITHADIAMETEROF10MMBAS,APLATINUMWIRE,ANDSILVER/SILVERCHLORIDEAG/AGCL/KCLWEREUSEDASAWORKING,ACOUNTER,ANDAREFERENCEELECTRODE,RESPECTIVELYFLOWAMPEROMETRICDETECTIONWASPERFORMEDINARADIALFLOWCELLMODEL113456,BASPBSWASFLOWEDATARATEOF170AL/MINAND50ALOFGOATANTIHUMANIGEIGGLABELEDWITHFCCHOANDFCCOOHWASFLOWEDINTOTHEFLOWCELLANDTHECURRENTRESPONSEWASMEASUREDUSINGAMPEROMETRICDETECTORMODELLC4C,BASRESULTSLABELINGOFIGGWITHFCCHOANDFCCOOHFORCONSTRUCTIONOFANELECTROCHEMICALIMMUNOASSAYPROBE,LABELINGOFGOATANTIHUMANIGEIGGWITHFCCHOANDFCCOOHWASINVESTIGATEDWHENFCCHOWASADDEDTOANIGGSOLUTIONATAMILDALKALINECONDITIONPH93,ITREACTEDWITHFREEAMINOGROUPSTOFORMANUNSTABLESCHIFFBASETYPECOMPOUNDAFTERTHISREACTIONSTEP,THISSCHIFFBASECOMPOUNDWASREDUCEDWITHSODIUMBOROHYDRIDETHENUMBEROFFERROCENEMOIETYBOUNDTOGOATANTIHUMANIGEIGGINCREASEDWITHANINCREASEDAMOUNTOFFCCHOINTHEREACTIONMIXTUREFIG2AMAXIMUMLABELINGNUMBEROFFERROCENEMOIETYWASOBTAINEDWHENFCCHOWASREACTEDATAMOLARRATIOOF1400IGGFCCHOINTHEREACTIONMIXTURETHEMAXIMUMMEANNUMBEROFFERROCENEMOIETYBOUNDTOGOATANTIHUMANIGEIGGWASEIGHTTHENUMBEROFFERROCENEMOIETYBOUNDTOGOATANTIHUMANIGEIGGWASREPRODUCIBLEWITHTHREEEXPERIMENTSANDAMEANNUMBEROFSEVENTOEIGHTFERROCENEMOIETYWEREBOUNDTOINDIVIDUALIGGATAHIGHERCONCENTRATIONOFFCCHOINTHEREACTIONMIXTUREWHENIGGANDFCCHOREACTEDATAMOLARRATIOOF1500,AGGREGATIONOFIGGWASOBSERVEDANDTHEBOUNDNUMBEROFFERROCENEMOIETYINTHEFILTRATEDECREASEDWHENLABELINGGOATANTIHUMANIGEIGGWITHFCCOOHVIAREACTIONOFSULFONHSANDEDC,THEMAXIMUMNUMBEROFBOUNDFERROCENEMOIETYWAS2TO3PERINDIVIDUALIGGITWASSHOWNTHATFCCHOCOULDEFFICIENTLYBINDTOIGGBYCHOOSINGANOPTIMUMPHANDREDUCINGSCHIFFBASEELECTROCHEMICALPROPERTIESOFIGGLABELEDWITHFCCHOANDFCCOOHCYCLICVOLTAMMOGRAMSOFGOATANTIHUMANIGEIGGLABELEDWITHFCCHOWASCARRIEDOUTINPBSATASCANRATEOF100MV/SUSINGAGLASSYCARBONELECTRODEATAPROTEINCONCENTRATIONOF34MG/MLOXIDATIONANDREDUCTIONPEAKSAPPEAREDAT390AND320MVVSAG/AGCL,RESPECTIVELYFIG3FORGOATANTIHUMANIGEIGGLABELEDWITHFCCOOHPROTEINCONCENTRATION,25MG/ML,ONLYASLIGHTOXIDATIONANDREDUCTIONPEAKAPPEAREDAT350AND280MVDATANOTSHOWNAHIGHERELECTROCHEMICALSIGNALCOULDBEOBTAINEDUSINGGOATANTIHUMANIGEIGGLABELEDWITHFCCHO,WHICHRETAINSAHIGHERNUMBEROFFERROCENEMOIETYOXIDATIONANDREDUCTIONPEAKSWEREREPRODUCIBLEWITHANERROROF15FORTHREEINDIVIDUALLYPREPAREDIGGLABELEDWITHFCCHOALSO,IGGLABELEDWITHFCCHOWASSTABLEFORATLEASTAFEWDAYSANDACHANGEINREDOXPOTENTIALORREDOXPEAKCURRENTWASNOTOBSERVEDBINDINGAFFINITYOFGOATANTIHUMANIGEIGGLABELEDWITHFCCHOTOANTIGENFIGURE4SHOWSTHEDEPENDENCEOFTHEMEANNUMBEROFBOUNDFERROCENEMOIETYPERIGGANDITSBINDINGAFFINITYFIGURE2RELATIONSHIPBETWEENTHEMEANNUMBEROFBOUNDFERROCENEMOIETYANDCONCENTRATIONOFFERROCENECARBOALDEHYDEINTHEREACTIONMIXTUREFORPREPARATIONOFIGGLABELEDWITHFERROCENECARBOALDEHYDE16BIOTECHNOLOGYANDBIOENGINEERING,VOL90,NO1,APRIL5,2005

簡介：中文中文3200字出處出處ICHIKAWAT,ERTURKSM,MOTOSUGIU,ETALHIGHBVALUEDIFFUSIONWEIGHTEDMRIINCOLORECTALCANCERJAMERICANJOURNALOFROENTGENOLOGY,2006,1871181184高B值彌散加權(quán)值彌散加權(quán)MRIMRI在結(jié)直腸癌中的應(yīng)用在結(jié)直腸癌中的應(yīng)用ICHIKAWAT,ERTURKSM,MOTOSUGIU,ETAL摘要摘要目的目的這篇文章的目的是評估高B值擴(kuò)散加權(quán)成像（DWMRI）檢測大腸腺癌的實(shí)用性。結(jié)論結(jié)論高B值DWMRI在檢測大腸腺癌方面具有很高的靈敏度和特異性檢測。關(guān)鍵詞癌癥；結(jié)腸；彌散加權(quán)成像；磁共振前言彌散加權(quán)MRI（DWMRI），在評估的惡性腫瘤變得越來越重要。人們普遍接受DW核磁共振成像是在使生物組織的非侵入性的特性基礎(chǔ)上，反映它們的水?dāng)U散性征，它能提供組織生物物理性質(zhì)方面的信息，如細(xì)胞的組織和密度，顯微組織及微循環(huán)。DW核磁共振成像已被廣泛用于在影像學(xué)，但其腹部內(nèi)的應(yīng)用卻受到大的生理運(yùn)動的阻礙，如呼吸，腸蠕動，血流量，及比擴(kuò)散更大的振幅等。高原直泰等提出了DWMRI的技術(shù)可能提供改進(jìn)的信號噪聲比（信噪比）的圖像，這些圖像的對比度，在黑色和白色圖像對比度特性密切類似PET逆轉(zhuǎn)。我們猜測，由于不同的健康和腫瘤組織的細(xì)胞結(jié)構(gòu)的不同，高B值DWMRI圖像，可以直接用于腫瘤的檢測。我們決定研究大腸腺癌，因?yàn)榻Y(jié)腸MRI有一定的挑戰(zhàn)，包括非實(shí)體性質(zhì)的器官，胃腸蠕動，和運(yùn)動腔內(nèi)的內(nèi)容物的干擾等。因此，我們在這個初步研究的目的是評估的有用的高B值DWMRI檢測大腸腺癌。材料及方法材料及方法患者患者2004年8月至2005年2月，為期6個月期間內(nèi)，在我們的機(jī)構(gòu)和兩個相關(guān)的醫(yī)院中共收集了33例患者（平均年齡59歲，范圍3369歲，15名女性，18名男性），并納入本研究。其中33例均經(jīng)結(jié)腸內(nèi)窺鏡證實(shí)大腸癌，病灶從20毫米到70毫米不等（平均33毫米）被發(fā)現(xiàn)在，病變位于直腸（14例），乙狀結(jié)腸（8例），橫結(jié)腸（N2），升結(jié)腸（8例），盲腸（1例）。另外選取同一時期15名結(jié)腸鏡檢為陰性的患者作為對照。所有大腸癌患者手術(shù)切除并證實(shí)診斷。所有患者和陰性對照組的病例在檢查前均行增強(qiáng)CT及MR。本研究施行前已經(jīng)我們的機(jī)構(gòu)審查委員會批準(zhǔn)且所有患者簽署知情同意書。MRIMRI序列及參數(shù)序列及參數(shù)研究為何高B值DWMR圖像上只有結(jié)腸腺癌，表現(xiàn)出強(qiáng)烈的信號強(qiáng)度，而健康的結(jié)腸則不是高信號的原因是一個具有挑戰(zhàn)性的問題，值得進(jìn)一步研究。然而，理論解釋可能構(gòu)造通過DW核磁共振成像良好的一般的假設(shè)。這是眾所周知的，擴(kuò)散即隨機(jī)的平移分子運(yùn)動，也被稱為布朗運(yùn)動。DWMRI是唯一的成像方法，該方法可以評估在體內(nèi)的擴(kuò)散過程。在細(xì)胞外和細(xì)胞內(nèi)的組件的組織的水分子的擴(kuò)散速度是不同的。細(xì)胞內(nèi)成分的擴(kuò)散速度相對較慢，這是因?yàn)榇嬖诩?xì)胞膜。因此，表觀擴(kuò)散系數(shù)（ADC），這是定量表達(dá)的組織中的擴(kuò)散特性，與胞外和胞內(nèi)組分的比例。他們往往會減少與增加組織細(xì)胞性或細(xì)胞密度。相反，細(xì)胞密度可能是腫瘤惡化的指標(biāo)LYNG以等報(bào)道腫瘤細(xì)胞高轉(zhuǎn)移能力增加。此外，在除了細(xì)胞膜，細(xì)胞內(nèi)的細(xì)胞骨架，細(xì)胞器，基質(zhì)型纖維和可溶性大分子在腫瘤的擴(kuò)散也限制。因此，擴(kuò)散的曲線迅速衰減或大型ADC值可能是典型的外大空間和小細(xì)胞性健康組織或良性的病理過程，而曲線衰減慢或小的ADC值可能表示惡性腫瘤或細(xì)胞過多。因此，DWMRI應(yīng)該可以敏感的鑒別病理組織特性。事實(shí)上，一些報(bào)道已經(jīng)指出各種惡性病變ADC值下降。然而，以前相關(guān)DWMR的研究并沒有直接視覺評估和報(bào)告這種技術(shù)的腹部病癥的診斷性能。雖然在本研究中所使用的技術(shù)主要基于DW核磁共振成像，事實(shí)上，DW核磁共振成像并沒有被經(jīng)常使用在臨床設(shè)置用于檢測大腸癌，但作為一個潛在的工具，用于治療監(jiān)測手段曾被提出過，建議使用的應(yīng)用程序是非定性得，但定量基于ADC測量。我們在這項(xiàng)研究中使用的高B值DWMRI技術(shù)與多個激勵和使用收購法無屏氣，提高信噪比。因?yàn)槠翚鈷呙钑r間的限制，不允許獲得足夠的SNR和多個激發(fā)，就不可以用來獲得作為多平面重建源圖像的彌散加權(quán)的薄層圖像。相反，運(yùn)動偽影的增加是可能存在的理論缺點(diǎn)，然而，在實(shí)踐中，被平均的運(yùn)動偽影期間多次激發(fā)的應(yīng)用使DW核磁共振成像和重建圖像變得不顯眼的運(yùn)動探測梯度。因此，圖像有更好的信噪比都達(dá)到了絕對變得無法計(jì)算，因?yàn)樾盘柶骄鵄DC值交換。我們的初步結(jié)果證明，高B值DWMRI對于檢測大腸癌的診斷表現(xiàn)為高靈敏度，30/33（91％）和特異性（100％，15/15），這種技術(shù)的其他優(yōu)點(diǎn)是，它是完全非侵襲性的，并不需要暴露于電離輻射或注入造影材料，并且不會引起病人的不適。此外，因?yàn)樗莵碜孕兄行У腄WMRI技術(shù)，高B值DWMRI并不需要運(yùn)營商提供先進(jìn)的技術(shù)技能或高成本的基礎(chǔ)設(shè)施投資，如回旋加速器PET等。高B值DWMRI的另一個優(yōu)點(diǎn)是，它是一個可以很容易地添加到MR檢查序列，因?yàn)樗恍枰粋€很短的掃描時間。在本研究中，我們沒有評估淋巴結(jié)轉(zhuǎn)移，我們的目的是評估高B值DWMRI檢測大腸腺癌的診斷能力。然而，在一些患者中，淋巴結(jié)在圖像上可以顯示?；谖覀兒喍痰挠跋駥W(xué)病理的相關(guān)性，大部分轉(zhuǎn)移淋巴結(jié)表現(xiàn)為高信號強(qiáng)度，所以可以被很好的檢測，但在一些患者健康的淋巴結(jié)也同樣顯示出很高的信號強(qiáng)度。關(guān)于特異性檢測淋巴結(jié)轉(zhuǎn)移，這種觀察可能是一個具有挑戰(zhàn)性的問題，有待進(jìn)一步研究。我們的研究有一定的局限性。首先，研究人口相對較少，我們的研究結(jié)果需要在更大范圍的臨床研究證實(shí)。其次，研究包括陰性的病例，但不包括其他良性疾病，如發(fā)炎性腸道疾病或良性腫瘤。因此，應(yīng)考慮在本研究中報(bào)道的靈敏度是相對的而不是絕對的?？傊?，根據(jù)我們的初步研究結(jié)果，高B值DWMRI可能是一個檢測大腸癌有用的工具，它顯示了較高的敏感性和特異性。然而，因?yàn)橐陨纤枋龅木窒扌裕孕枰M(jìn)一步的大樣本研究來支持我們的調(diào)查結(jié)果。

簡介：中文中文4900字出處出處RUSCAN,MONTICELLISMIR146AINIMMUNITYANDDISEASEJMOLECULARBIOLOGYINTERNATIONAL,2011,17免疫和疾病中的免疫和疾病中的MIR146MIRNA分子是一類在細(xì)胞中可以影響各種生物功能的分子。已經(jīng)證實(shí)他們與癌癥、病毒感染和免疫性疾病相關(guān)，近些年，還發(fā)現(xiàn)它們成為免疫反應(yīng)的重要調(diào)節(jié)者。尤其是MIR146A迅速成為一種重要的分化調(diào)節(jié)者，同樣在先天免疫和獲得性免疫中有很重要的作用。鑒于它在調(diào)節(jié)細(xì)胞功能方面的重要作用，MIRNA146A在各種腫瘤中被發(fā)現(xiàn)表達(dá)失調(diào)也不足為奇。這篇文章，我們主要總結(jié)了近些年對MIR146A在先天免疫和獲得性免疫反應(yīng)及疾病中的作用的進(jìn)展。1介紹介紹MIRNA代表了所有的細(xì)胞的普遍特征，因?yàn)樗鼈冋{(diào)節(jié)了細(xì)胞轉(zhuǎn)錄過程中的大部分。至今，672個鼠的MIRNA分子和1048個人的MIRNA已經(jīng)在MIRBASE數(shù)據(jù)庫中被描述（HTTP//WWWMIRBASEORG/,RELEASESEPT2010其中每個MIRNA潛在地調(diào)節(jié)了成百上千的目標(biāo)基因，這顯著地?cái)U(kuò)展了調(diào)節(jié)模式。然而一些MIRNA表達(dá)廣泛，其他的僅僅抑制有限發(fā)展的階段、組織或者細(xì)胞特異的類型。類似于一些哺乳細(xì)胞的類型，細(xì)胞免疫系統(tǒng)依賴MIRNA來調(diào)節(jié)譜系的定型、增殖、遷移和分化。在大部分的情況下，這些活動都是十分和諧，在廣泛的表達(dá)和細(xì)胞特定的MIRNA類型方面。當(dāng)MIRNA的表達(dá)被改變時MIRNA在調(diào)節(jié)分化和免疫細(xì)胞功能方面的重要性就通過表型干擾表現(xiàn)得尤為突出。MIRNA在修飾免疫反應(yīng)的重要作用，可能任何MIRNA表達(dá)的失調(diào)都會導(dǎo)致自身免疫性疾病、慢性炎癥和惡性腫瘤的發(fā)生。事實(shí)上，已經(jīng)證明人類的一些疾病和MIRNA的表達(dá)失調(diào)有關(guān)，MIRNA的功能可以充當(dāng)癌基因和抑癌基因。MIR146A最近已經(jīng)報(bào)道成為在先天和獲得性免疫中重要的細(xì)胞分化和功能的調(diào)節(jié)者。在此，我們總結(jié)了近來關(guān)于MIR146A在免疫反應(yīng)和疾病中作用的理解（見表1）。2什么是什么是MIRNA（此部分未翻譯，和之前的文章基本重復(fù)）（此部分未翻譯，和之前的文章基本重復(fù)）MIRNA是一種非編碼小RNA分子（長度2025核苷酸），參與轉(zhuǎn)錄后基因的調(diào)控。它們最初為初級MIRNAPREMIRNA，這會在細(xì)胞核內(nèi)通過微處理復(fù)合體形成前體發(fā)夾結(jié)構(gòu)（PREMIRNA），這種復(fù)合體發(fā)函RNASEIII酶DROSHA。3MIRNA146A在獲得性免疫反應(yīng)中在獲得性免疫反應(yīng)中雖然獲得性免疫應(yīng)答反應(yīng)失調(diào)會導(dǎo)致自身免疫性疾病和慢性炎癥疾病，但是能有效地消滅病原體的感染。獲得性免疫反應(yīng)的發(fā)展和進(jìn)步對于侵入的病原體會形成一系列精密的反應(yīng)步驟，包括免疫細(xì)胞的活化增殖和隨后遷移到炎癥部位。最初的跡象表明MIRNA參與調(diào)節(jié)免疫細(xì)胞的分化，這已經(jīng)被陳和他的同事們用MIR181在造血干細(xì)胞中特異性的表達(dá)證明。在造血干細(xì)胞中的易位表達(dá)導(dǎo)致了部分B細(xì)胞系在體外誘導(dǎo)分化和大鼠的模型中的增加。接下來的開創(chuàng)性的研究，證明MIRNA是控制免疫細(xì)胞分化和功能重要的組成部分。當(dāng)與抗原結(jié)合后，原始CD4T細(xì)胞升高了T細(xì)胞亞群（TH1，TH2，TH17，TREGS，濾泡輔助性T細(xì)胞）根據(jù)它們各自在宿主防御中的功能。最初的表達(dá)鑒定了MIRNA的表達(dá)譜在不同的T細(xì)胞亞群和部分分化階段。T細(xì)胞特異性切酶顯示在T細(xì)胞的發(fā)展中需要MIRNA通路，同時T細(xì)胞亞群的分化也需要。事實(shí)上，缺乏DICER酶的T細(xì)胞表現(xiàn)出向TH1亞群的分化增加，同時向TH2減少。根據(jù)復(fù)雜的基因調(diào)控網(wǎng)絡(luò)，增殖的T細(xì)胞比靜止的T細(xì)胞表達(dá)更短的3‘UTR，使得這些MRNA更不易被MIRNA調(diào)控由于缺失MIRNA結(jié)合位點(diǎn)。最終，個別的MIRNA對T細(xì)胞分化和功能有重要的作用。例如，MIR181A，面對病毒的入侵時免疫細(xì)胞使用各個層次的負(fù)向調(diào)節(jié)以防免疫應(yīng)答不受控制，這種調(diào)節(jié)機(jī)制也可以被病毒使用為了逃離免疫監(jiān)視。發(fā)現(xiàn)MIR146A在調(diào)節(jié)皰疹性口腔炎病毒（VSV）的感染時有作用。在巨噬細(xì)胞中，VSV的感染上調(diào)了MIR146A的表達(dá)，導(dǎo)致負(fù)向調(diào)節(jié)VSV觸發(fā)的I型IFN通過下調(diào)TRAF6，IRAK1和IRAK2，因此促進(jìn)了VSV在巨噬細(xì)胞中的復(fù)制。作者提出了一種模式，VSV的感染首先被RIGI（視黃酸誘導(dǎo)基因蛋白I）感受到，這反過來抑制I型IFN的產(chǎn)生來對抗VSV的感染。同時，VSV的感染上調(diào)了MIR146A的表達(dá)，通過破壞RIGI信號抑制了先天免疫應(yīng)答。EBV（人類皰疹病毒4）感染過超過90的全世界的人口。EV病毒感染會導(dǎo)致惡性腫瘤，包括BURKITT’S和霍奇金淋巴瘤。LMP1潛伏膜蛋白是EV病毒編碼的主要的癌基因產(chǎn)物，它可以活化轉(zhuǎn)錄因子如NFKB和AP1，由此來控制宿主細(xì)胞、調(diào)節(jié)細(xì)胞分化、遷移和凋亡的過程。通過它的這種活化轉(zhuǎn)錄因子的能力，LMP1也能誘導(dǎo)細(xì)胞中MIRNA的表達(dá)，其中最顯著的是MIR146A，因此在感染EB病毒后對細(xì)胞的永生化和腫瘤發(fā)生有幫助。6MIR146和癌癥和癌癥癌癥是一系列復(fù)雜過程的結(jié)果，是一些基因各種順序改變的積累，包括編碼MIRNA。既然MIRNA參與保持基因平衡的任務(wù)，那么它也可以決定細(xì)胞的命運(yùn)，它們的失調(diào)潛在地會減弱這種平衡，因此可能導(dǎo)致癌癥的發(fā)生、發(fā)展。事實(shí)上，MIRNA表達(dá)譜中已經(jīng)發(fā)現(xiàn)在一些癌癥中MIRNA的表達(dá)被顯著地改變了。關(guān)于MIR146A可能參與癌癥發(fā)展的初步證據(jù)來自MIR146A在PTC（甲狀腺乳頭狀癌）樣本中被上調(diào)的研究與未受影響的甲狀腺組織相比。有趣的是，一組5個MIRNA，包括MIR221，MIR222，MIR146對區(qū)分PTC和正常甲狀腺組織是足夠的了。在免疫設(shè)置中進(jìn)行同樣的觀察，MIR146A/B在代謝旺盛的人類乳腺癌細(xì)胞株MDAMB231中的過表達(dá)顯著下調(diào)了IRAK1和TRAF6的表達(dá)，負(fù)調(diào)節(jié)了NFKB的活性。在功能上，這導(dǎo)致與控制組的細(xì)胞相比這些細(xì)胞的侵襲和轉(zhuǎn)移的能力明顯受損。這些發(fā)現(xiàn)說明MIR146在乳腺癌細(xì)胞中不僅是NFKB的負(fù)向調(diào)節(jié)者，而且說明調(diào)節(jié)MIR146的水平或許可以潛在地抑制乳腺癌的轉(zhuǎn)移。用相同的路線，在眾多的MIRNA中發(fā)現(xiàn)與正常宮頸組織相比MIR146A在宮頸癌組織中上調(diào)。當(dāng)引入細(xì)胞株時，發(fā)現(xiàn)MIR146A促進(jìn)了細(xì)胞增殖。雖然這種增加增殖的分子機(jī)制已經(jīng)被研究，但是這些發(fā)現(xiàn)說明MIR146A可能與宮頸癌的發(fā)生有關(guān)。另一種癌癥激素難治性前列腺癌（HRPC），MIR146A的水平減少了，與雄激素敏感性的非癌癥上皮細(xì)胞相比。在這方面，MIR146A扮演腫瘤抑制的角色，減少了它的靶點(diǎn)ROCK1的水平，ROCK1是參與HRPC轉(zhuǎn)化的一個關(guān)鍵激酶。因此，強(qiáng)化MIR146A的表達(dá)可以減少ROCK1蛋白的水平、細(xì)胞分化、侵襲和向人單層骨髓上皮細(xì)胞的轉(zhuǎn)移。同樣，MIR146A在胰腺癌細(xì)胞中水平較低與正常人胰腺細(xì)胞相比。MIR146A的表達(dá)通過下調(diào)EGFR（表皮生長因子受體）和IRAK1抑制了胰腺癌細(xì)胞的侵襲能力。最后，最近的一項(xiàng)研究說明用DZ（二氮嗪）治療骨髓源性的間充質(zhì)肝細(xì)胞（MSCS）顯著地增加了MIR146A的表達(dá)，促進(jìn)了細(xì)胞的存活。此外，通過反義抑制劑MIR146A水平的下調(diào)消除了DZ誘導(dǎo)的細(xì)胞保護(hù)作用。這說明MIR146A在MSC（骨髓基質(zhì)細(xì)胞）存活中有重要作用。7基因多態(tài)性和轉(zhuǎn)錄后的修飾基因多態(tài)性和轉(zhuǎn)錄后的修飾基因多態(tài)性影響MIRNA的表達(dá)、成熟或者M(jìn)RNA的識別可能對增加腫瘤的風(fēng)險(xiǎn)成為重要的決定因素。事實(shí)上，最近KIT癌基因3‘UTR遺傳變異被描述，這導(dǎo)致了MIR221種子區(qū)的不匹配，伴隨增加黑色素瘤的風(fēng)險(xiǎn)。至于MIR146A，單核苷酸多態(tài)性在前MIR146A的傳遞鏈上被發(fā)現(xiàn)。罕見的C等位基因降低了和前體MIR146A合成的過程，降低了PREMIR146A和成熟MIR146A的水平，解鎖了它的靶基因，包括TRAF6和IRAK1。一項(xiàng)與PTC患者有關(guān)的研究，GC雜合狀態(tài)會增加獲得性PTC的風(fēng)險(xiǎn)，但是兩個純合子狀態(tài)

簡介：SHORTCOMMUNICATIONMOLECULARANDCLINICALDIFFERENCESBETWEENADENOCARCINOMASOFTHEESOPHAGUSANDOFTHEGASTRICCARDIAPHILIPPETANIERE,GHISLAINEMARTELPLANCHE,DANIELAMAURICI,CATHERINELOMBARDBOHAS,?JEANYVESSCOAZEC,?RUGGEROMONTESANO,FRANC?OISEBERGER,?ANDPIERREHAINAUTFROMTHEINTERNATIONALAGENCYFORRESEARCHONCANCERTHEFE′DE′RATIONDESSPE′CIALITE′SDIGESTIVES?ANDTHELABORATOIRED’ANATOMIEPATHOLOGIQUE,?HO?PITALEDOUARDHERRIOT,LYON,FRANCEADENOCARCINOMAOFTHEESOPHAGUSADCEWITHBARRETT’SMUCOSAANDADENOCARCINOMAOFTHECARDIAADCCAREOFTENREPORTEDASASINGLEPATHOLOGICALENTITYINTHISSTUDYWEHAVEUSEDSTRICTANATOMICALPATHOLOGICALCRITERIATODISTINGUISHBETWEENTHESETWOLESIONSANDWEHAVEINVESTIGATEDTHEIRDIFFERENCESINTP53MUTATIONS,MDM2GENEAMPLIFICATION,ANDCYTOKERATINEXPRESSIONDNAWASEXTRACTEDFROMTHETUMORAREASOFFORMALINFIXED,PARAFFINEMBEDDEDSECTIONSIN26ADCCAND28ADCEPATIENTSTP53MUTATIONSWEREDETECTEDBYTEMPORALTEMPERATUREGRADIENTELECTROPHORESISANDIDENTIFIEDBYSEQUENCINGMDM2AMPLIFICATIONWASASSESSEDBYDIFFERENTIALPOLYMERASECHAINREACTIONTHEEXPRESSIONOFCYTOKERATINS4,7,AND13WASEXAMINEDBYIMMUNOHISTOCHEMISTRYINADCC,THEMALETOFEMALERATIOWAS181,COMPAREDTO271INADCEFIVEADCCPATIENTSHADAHISTORYOFOTHERNEOPLASMS,COMPAREDTOONLYONEADCEPATIENTTHETWOTYPESOFTUMORDIFFEREDINTHEPREVALENCEOFTP53MUTATIONS31INADCCAND50INADCEANDOFMDM2GENEAMPLIFICATION19INADCCAND4INADCE,ANDINTHEPATTERNOFEXPRESSIONOFCYTOKERATIN7POSITIVEIN100OFADCEANDIN41OFADCCANDCYTOKERATIN13POSITIVEIN81OFADCEANDIN365OFADCCADCEANDADCCDIFFERINTHEIRCLINICALCHARACTERISTICS,INTHEPREVALENCEOFTP53MUTATIONSANDMDM2AMPLIFICATIONS,ANDINTHEPATTERNSOFCYTOKERATINEXPRESSIONTHESERESULTSSUPPORTTHENOTIONTHATADCCANDADCEAREDISTINCTPATHOLOGICALENTITIESAMJPATHOL2001,15833–40THROUGHOUTTHEPAST20YEARS,THEINCIDENCEOFTUMORSOFTHEESOPHAGOGASTRICJUNCTIONHASINCREASEDATARATEOF5TO10PERYEARINTHEUNITEDSTATESANDSEVERALWESTERNEUROPEANCOUNTRIES1THEREASONSFORTHISINCREASEAREPRIMARILYUNKNOWNTUMORSOFTHEESOPHAGOGASTRICJUNCTIONINCLUDETWOMAJORTYPESOFADENOCARCINOMAADENOCARCINOMASOFTHEESOPHAGUSADCEANDADENOCARCINOMASOFTHECARDIAADCCADCEOCCURINTHEDISTALPARTOFTHEESOPHAGUSANDDEVELOPFROMBARRETT’SMUCOSA,AGLANDULARMETAPLASIAOFTHESQUAMOUSEPITHELIUMTHATCANVARYINHEIGHTFROMAFEWMILLIMETERSTOAFEWCENTIMETERSTHEREISEVIDENCETHATTHEMETAPLASTICGLANDULARCELLSAREHYBRIDCELLS,EXPRESSINGCYTOKERATINSCKSOFBOTHSQUAMOUSCK4AND13ANDGLANDULARCK8AND19ORIGIN2ANDHAVINGULTRASTRUCTURALFEATURESOFBOTHSQUAMOUSANDGLANDULARCELLS3FURTHERMORE,THEYHAVEBEENSHOWNTOCONSTANTLYEXPRESSCK7,INCONTRASTTOINTESTINALMETAPLASTICCELLSOFTHECARDIAMUCOSA,WHICHNEVERDO4BARRETT’SMUCOSAISOFTENASSOCIATEDWITHCHRONICGASTROESOPHAGEALACIDREFLUXHOWEVER,ITCANALSOOCCURINCOMBINATIONWITHCHRONICBILIARYALKALINEREFLUXASWELLASINTHEABSENCEOFREFLUX5FACTORSPREDISPOSINGTOBARRETT’SMUCOSAARENOTWELLDOCUMENTEDRECENTEVIDENCESUGGESTSTHATEXPRESSIONOFCERTAINPOLYMORPHICFORMSOFGLUTATHIONESTRANSFERASEP1MAYBEAGENETICSUSCEPTIBILITYFACTORFORDEVELOPINGBARRETT’SMUCOSA6BARRETT’SMUCOSAISAVERYCOMMONLESIONTHATISTHOUGHTTOOCCURIN?10OFTHEGENERALPOPULATIONINTHEUNITEDSTATESANDISASSOCIATEDWITHA10FOLDINCREASEINTHERISKOFDEVELOPINGADCE7THECARDIAISTHEANATOMICALREGIONCORRESPONDINGTOTHETRANSITIONBETWEENESOPHAGUSANDSTOMACHITCANNOTBEIDENTIFIEDATTHEMACROSCOPICLEVELATTHEMICROSCOPICLEVEL,THECARDIAISCHARACTERIZEDBYATHINMUCOSAWITHCLEARGLANDULARCELLS,WITHOUTANYACIDSECRETINGCELLSITRANGESINHEIGHTFROM1TO5MM,WITHANINCREASEINSIZEWITHAGETHETERM“ADCC”APPLIESTOTUMORSLOCATEDACCEPTEDFORPUBLICATIONSEPTEMBER14,2000ADDRESSREPRINTREQUESTSTOPHAINAUT,PHD,GROUPOFMOLECULARCARCINOGENESIS,IARC,150COURSALBERTTHOMAS,69372LYONCEDEX08,FRANCEEMAILHAINAUTIARCFRAMERICANJOURNALOFPATHOLOGY,VOL158,NO1,JANUARY2001COPYRIGHT?AMERICANSOCIETYFORINVESTIGATIVEPATHOLOGY33ANTIBODY,CLONEAE1/AE3,1/100DAKO,COPENHAGEN,DENMARK,ANDCK13MONOCLONALANTIBODY,CLONEKS1A3,1/50NOVOCASTRALABORATORIESLTDINCUBATIONWITHTHERELEVANTSECONDARYANTIBODIESEITHERANTIMOUSEORANANTIRABBITBIOTINYLATEDIGG,1/200,VECTASTAINELITEABCKITVECTORLABORATORIESINCFOR30MINUTESATROOMTEMPERATUREWASFOLLOWEDBYSTREPTAVIDINPEROXIDASE1/50,30MINUTESAT37°CPEROXIDASEACTIVITYWASDETECTEDWITHADIAMINOBENZIDINEBASEDDETECTIONKITVECTORLABORATORIES,INCANDSECTIONSWERECOUNTERSTAINEDWITHMAYER’SHEMATOXYLINBEFOREDEHYDRATIONANDMOUNTINGTP53MUTATIONANALYSISTP53EXONS4TO9WEREANALYZEDBYTEMPORALTEMPERATUREGRADIENTELECTROPHORESISUSINGTHEDGENESYSTEMBIORAD,RICHMOND,CAANDTHEPRIMERSDESCRIBEDBYHAMELINANDCOLLEAGUES20EXONS5,7,AND8ANDGULDBERGANDCOLLEAGUES21EXONS4,6,AND9DNAWASAMPLIFIEDINADNATHERMOCYCLERPERKINELMER,NORWALK,CTINA50?LREACTIONMIXTURECONTAINING5?LOFGENOMICDNA,20PMOLOFSENSEANDANTISENSEPRIMERS,200?MOL/LOFEACHDNTP,1?AMPLIFICATIONBUFFER,1XQSOLUTIONAND05?L25UOFTAQPOLYMERASEHOTSTARTAQDNAPOLYMERASEQIAGEN,HILDEN,GERMANYPOLYMERASECHAINREACTIONPCRCONDITIONSWERE15MINUTESAT95°CFOLLOWEDBY35CYCLESAT95°C1MINUTE,56°CEXONS5PROXIMAL,8,AND9OR62°CEXONS4A,4B,5DISTAL,6,AND71MINUTE,72°C90SECONDSTHEREACTIONWASENDEDBYA10MINUTESEXTENSIONAT72°CHETERODUPLEXFORMATIONWASINDUCEDBYDENATURATIONFOR10MINUTESAT98°C,FOLLOWEDBY30MINUTESATTHERESPECTIVEANNEALINGTEMPERATURE56°COR62°CTEMPORALTEMPERATUREGRADIENTELECTROPHORESISWASRUNAT130VATTEMPERATURESOPTIMIZEDFOREACHDNAFRAGMENTEXON4P,4D,6,AND958TO70°CEXON5P56TO70°CEXON5D63TO70°CEXON759TO70°CEXON853TO67°CANEGATIVECONTROLWILDTYPESAMPLEANDPOSITIVECONTROLKNOWNMUTANTWEREINCLUDEDINEACHANALYSISSAMPLESTHATSHOWEDADDITIONALAND/ORABNORMALBANDSWEREREAMPLIFIEDFROMGENOMICDNAANDASECONDTEMPORALTEMPERATUREGRADIENTELECTROPHORESISWASPERFORMEDIFCONFIRMED,MUTANTALLELESWERECUTFROMTHISSECONDGEL,REAMPLIFIEDUSINGTHESAMEPRIMERS,ANDANALYZEDBYDIRECTSEQUENCINGAFTERASYMMETRICPCRAMPLIFICATIONSASPREVIOUSLYDESCRIBED20,22TWOCASESWITHPOSITIVEP53IMMUNOSTAINING?50DIDNOTSHOWREPRODUCIBLEPATTERNSOFABNORMALBANDSINTEMPORALTEMPERATUREGRADIENTELECTROPHORESISCASE12,TABLE1,ANDCASE3,TABLE2INTHESETWOCASES,MRNAWASISOLATEDFROMFROZENBIOPSIES,ANDCDNAWASPREPAREDANDTESTEDUSINGTHEYEASTFUNCTIONALASSAYASDESCRIBEDBYFLAMANETAL23POSITIVECOLONIESWERESEQUENCEDUSINGTHEABIPRISM310GENETICANALYZERPERKINELMERBIOSYSTEMS,FOSTERCITY,CAANALYSISOFMDM2GENEAMPLIFICATIONDIFFERENTIALPCRWASPERFORMEDASPREVIOUSLYDESCRIBED24WITHTHEFOLLOWINGMODIFICATIONS5?LOFTEMPLATEDNAWASAMPLIFIEDIN50?LOFAREACTIONMIXTURECONTAINING20PMOLEACHOFSENSEANDANTISENSEPRIMERSFORMDM2ANDFORTHEDOPAMINED2RECEPTORGENEDRD2USEDASAREFERENCE,200?MOL/LOFEACHDNTP,1?AMPLIFICATIONBUFFER,1XQSOLUTIONAND05?L25UOFHOTSTARTAQDNAPOLYMERASEQIAGENPCRCONDITIONSWERE15MINUTESAT95°C,FOLLOWEDBY27CYCLESAT95°CFOR45SECONDS,55°CFOR45SECONDS,AND72°CFOR1MINUTEWITHAFINALEXTENSIONAT72°CFOR5MINUTESTHEPRIMERSWEREASFOLLOWS5?GAGGGCTTTGATGTTCCTGA3?SENSEAND5?GCTACTAGAAGTTGATGGC3?ANTISENSEFORMDM2,AND5?CCACTGAATCTGTCCTGGTATG3?SENSEAND5?GTGTGGCATAGTAGTTGTAGTGG3?ANTISENSEFORHUMANDRD2PCRPRODUCTSWEREELECTROPHORESEDON75POLYACRYLAMIDEGELSSTAINEDWITHETHIDIUMBROMIDE,PHOTOGRAPHED,ANDTHEFILMSWEREANALYZEDBYSCANNINGDENSITOMETRYGS670BIORAD,HERCULES,CAANMDM2/DRD2RATIOOF25ORABOVEWASREGARDEDASINDICATIVEOFMDM2AMPLIFICATIONANDARATIOBETWEEN2AND25WASREGARDEDASCOMPATIBLEWITHMDM2AMPLIFICATIONSTATISTICALEVALUATIONSFREQUENCYTABLESOFINDEPENDENTVARIABLESWEREEVALUATEDFORSTATISTICALSIGNIFICANCEBYPEARSON’SCHISQUARETESTRESULTSCLINICALANDINDIVIDUALCHARACTERISTICSOFTHEPATIENTSTWENTYSIXCASESOFADCCAND28CASESOFADCEWERECOLLECTEDBETWEEN1995AND1999TABLES1AND2INONECASE,ATUMORWASCLASSIFIEDASADCEONTHEBASISOFTHEPREVIOUSDIAGNOSISONBIOPSYOFABARRETT’SMUCOSATHATWASNOLONGERDETECTABLEATSURGERYNONEOFTHEPATIENTSHADRECEIVEDCHEMOTHERAPYORRADIOTHERAPYBEFOREBIOPSYORSURGERYTHEMEANAGEOFPATIENTSWAS621?136YEARSRANGE,25TO82YEARSFORADCCAND681?9YEARSRANGE,50TO82YEARSFORADCETHEGROUPOFADCCPATIENTSINVESTIGATEDINCLUDED17MENANDNINEWOMENMALE/FEMALERATIOOF065,WHEREASTHEADCEPATIENTSWEREALMOSTEXCLUSIVELYMALES27MALESANDONEFEMALEMALE/FEMALERATIO097DESPITETHESHORTFOLLOWUPPERIODFORSOMEOFTHEPATIENTS,MEDICALRECORDSREVEALEDTHATFIVEOFTHEADCCPATIENTSHADADDITIONALTUMORSTHREEWOMENDEVELOPEDABREASTADENOCARCINOMAEITHERBEFORE19YEARS,PATIENT422YEARS,PATIENT46ORAFTER3YEARS,PATIENT33DIAGNOSISOFADCCONEOFTHESEPATIENTSPATIENT46ALSODEVELOPEDAMALIGNANTMELANOMA10YEARSBEFOREADCCONEMANPATIENT38HADANADENOCARCINOMAOFTHEINTESTINE10YEARSBEFOREADCCANDANOTHERPATIENT29PRESENTEDAPLEOMORPHICADENOMAOFTHEPAROTID5YEARSBEFOREADCCAMONGTHEADCEPATIENTS,ONLYONEPATIENTHADAHISTORYOFAPREVIOUSCANCERASQUAMOUSCELLCARCINOMAOFHEADANDNECKINAMALEPATIENTWHOWASAHEAVYSMOKERANDESOPHAGEALANDGASTRICCARDIAADENOCARCINOMAS35AJPJANUARY2001,VOL158,NO1

簡介：點(diǎn)擊查看更多[雙語翻譯]外文翻譯-醫(yī)學(xué)圖像水印基準(zhǔn)中英全精彩內(nèi)容。

簡介：AJR187,JULY2006181AJR2006187181–1840361–803X/06/1871–181?AMERICANROENTGENRAYSOCIETYMEDICALIMAGINGACENTURYOFMEDICALIMAGINGACENTURYOFICHIKAWAETALHIGHBVALUEDIFFUSIONWEIGHTEDMRIINCOLORECTALCANCERGASTROINTESTINALIMAGINGTECHNICALINNOVATIONHIGHBVALUEDIFFUSIONWEIGHTEDMRIINCOLORECTALCANCERTOMOAKIICHIKAWA1SUKRUMEHMETERTURK2,3UTAROUMOTOSUGI1HIRONOBUSOU1HIROSHIIINO4TSUTOMUARAKI1HIDEKIFUJII4ICHIKAWAT,ERTURKSM,MOTOSUGIU,ETALKEYWORDSCANCER,COLON,DIFFUSIONWEIGHTEDMRI,MRIDOI102214/AJR051005RECEIVEDJUNE11,2005ACCEPTEDAFTERREVISIONSEPTEMBER13,20051DEPARTMENTOFRADIOLOGY,YAMANASHIUNIVERSITY,SHIMOKATO,JAPAN2BRIGHAMANDWOMEN’SHOSPITAL,HARVARDMEDICALSCHOOL,RADIOLOGYSUITE,C/OONEBRIGHAMCIRCLE,1620TREMONTST,BOSTON,MA02120ADDRESSCORRESPONDENCETOSMERTURKMEHMETERTURKSUPERONLINECOM3DEPARTMENTOFRADIOLOGY,SISLIETFALTRAININGANDRESEARCHHOSPITAL,ISTANBUL,TURKEY4FIRSTDEPARTMENTOFSURGERY,UNIVERSITYOFYAMANASHI,SHIMOKATO,JAPANOBJECTIVETHEPURPOSEOFTHISARTICLEISTOEVALUATETHEUSEFULNESSOFHIGHBVALUEDIFFUSIONWEIGHTEDMRIDWMRIINTHEDETECTIONOFCOLORECTALADENOCARCINOMACONCLUSIONHIGHBVALUEDWMRIALLOWSDETECTIONOFCOLORECTALADENOCARCINOMAWITHAHIGHSENSITIVITYANDSPECIFICITYIFFUSIONWEIGHTEDMRIDWMRIISBECOMINGINCREASINGLYIMPORTANTINTHEASSESSMENTOFMALIGNANTTUMORS1,2ITISGENERALLYACCEPTEDTHATDWMRIENABLESNONINVASIVECHARACTERIZATIONOFBIOLOGICTISSUESONTHEBASISOFTHEIRWATERDIFFUSIONPROPERTIES3ITPROVIDESINFORMATIONABOUTTHEBIOPHYSICALPROPERTIESOFTISSUESSUCHASCELLORGANIZATIONANDDENSITY,MICROSTRUCTURE,ANDMICROCIRCULATION4DWMRIISWIDELYUSEDINNEUROIMAGING5,BUTITSAPPLICATIONWITHINTHEABDOMENISHINDEREDBYTHEPRESENCEOFBULKPHYSIOLOGICMOTIONSUCHASRESPIRATION,PERISTALSIS,ANDBLOODFLOW,INWHICHORDERSOFMAGNITUDEAREGREATERINAMPLITUDETHANTHATOFDIFFUSION6TAKAHARAETAL7PROPOSEDADWMRITECHNIQUETHATMIGHTPROVIDEIMAGESWITHIMPROVEDSIGNALTONOISERATIOSSNRSREVERSALOFTHECONTRASTOFTHESEIMAGESRESULTEDINBLACKANDWHITEIMAGESWITHCONTRASTCHARACTERISTICSCLOSELYRESEMBLINGTHOSEOFPETWEHYPOTHESIZEDTHATHIGHBVALUEDWMRIIMAGESCOULDBEDIRECTLYUSEDFORTUMORDETECTIONBECAUSEOFTHEDIFFERENTCELLULARSTRUCTURESOFHEALTHYANDNEOPLASTICTISSUESWEDECIDEDTOSTUDYCOLORECTALADENOCARCINOMABECAUSEOFTHEGENERALCHALLENGESOFCOLONICMRI,INCLUDINGTHENONSOLIDNATUREOFTHEORGAN,PERISTALSIS,ANDMOVEMENTOFTHEINTRALUMINALCONTENTSTHUS,OURAIMINTHISPRELIMINARYSTUDYWASTOEVALUATETHEUSEFULNESSOFHIGHBVALUEDWMRIINTHEDETECTIONOFCOLORECTALADENOCARCINOMAMATERIALSANDMETHODSPATIENTSDURINGAPERIODOF6MONTHSBETWEENAUGUST2004ANDFEBRUARY2005,33CONSECUTIVEPATIENTSMEANAGE,59RANGE,33–69YEARS15WOMEN,18MENWITH33ENDOSCOPICCOLONOSCOPICALLYPROVENCOLORECTALCANCERSRANGINGFROM20TO70MMMEAN,33MMWEREFOUNDINOURINSTITUTIONANDTWORELATEDHOSPITALSANDWEREINCLUDEDINTHISSTUDYTHELESIONSWERELOCATEDINTHERECTUMN14,SIGMOIDCOLONN8,TRANSVERSECOLONN2,ASCENDINGCOLONN8,ANDCECUMN1ANOTHER15PATIENTSWHOUNDERWENTENDOSCOPICCOLONOSCOPYDURINGTHESAMEPERIODWITHNEGATIVERESULTSWEREINCLUDEDASCONTROLSALLPATIENTSWITHCOLORECTALCANCERFINALLYUNDERWENTSURGICALRESECTIONANDTHEDIAGNOSESWERECONFIRMEDCONTRASTENHANCEDCTWASPERFORMEDINALLPATIENTSANDNEGATIVECASESINTHECONTROLGROUPBEFOREMREXAMINATIONSINFORMEDCONSENTWASOBTAINEDFROMALLPATIENTSBEFOREPARTICIPATIONINTHESTUDY,WHICHWASAPPROVEDBYOURINSTITUTIONALREVIEWBOARDMRIPROTOCOLANDPARAMETERSMRIUSEDACOMBINATIONOFACOMMERCIALLYAVAILABLE15TSUPERCONDUCTINGMRUNITANDABODYCOILSIGNAECHOSPEED,GEHEALTHCAREFIRST,BREATHHOLD,CORONALT1WEIGHTEDMRIMAGESWITHGRADIENTECHOSEQUENCESWEREOBTAINEDTOCONFIRMTHEOPTIMALSCANRANGETHEN,AXIALT1WEIGHTEDANDRESPIRATORYTRIGGERED,FASTSPINECHOT2WEIGHTEDMRITR/TE,2,000–4,000/80,ANDHIGHBVALUEDWMRIWEREPERFORMEDINALLPATIENTSINCLUDINGTHECONTROLGROUPTHEPATIENTSDIDNOTUNDERGOANYPREPARATIONSUCHASBOWELCLEANSINGBEFORETHEEXAMINATIONSHIGHBVALUEDWMRIMAGESWEREOBTAINEDWITHOUTBREATHHOLDINGDURINGTHEACQUISITIONDETAILEDPARAMETERSFORHIGHBVALUEDWMRIWERESEQUENCESINGLESHOTSPINECHOECHOPLANARSEEPIFATSUPPRESSIONTECHNIQUE,CHEMICALSHIFTSELECTIVETECHNIQUESCANDIRECTION,AXIAL,BVALUE,ZEROAND1,000S/MM2TR/TE/INVERSIONTIMETI,8,000–10,000/732–734/70MATRIX,12864SLICEDDOWNLOADEDFROMWWWAJRONLINEORGBY59474389ON05/15/13FROMIPADDRESS59474389COPYRIGHTARRSFORPERSONALUSEONLYALLRIGHTSRESERVEDHIGHBVALUEDIFFUSIONWEIGHTEDMRIINCOLORECTALCANCERAJR187,JULY2006183WITHOUTREFERRINGTOANYOTHERMRIMAGESMIPIMAGESWEREEVALUATEDWITHTHEROTATIONALCINEMODETOGETHERWITHTHEAXIALSOURCEIMAGESINDIFFERENTWINDOWSONDIAGNOSTICMONITORSEACHREVIEWERGRADEDTHEPRESENCEORABSENCEOFLESIONSONA5POINTCONFIDENCESCALEBASEDONTHESTRENGTHANDTHEAPPEARANCEOFDARKSIGNALSONHIGHBVALUEDWMRIMAGESASFOLLOWS1DEFINITELYABSENTNOSIGNAL2PROBABLYABSENTNONLOCALIZED,MILDTOMODERATESIGNAL3UNDETERMINEDLOCALIZED,MILDTOMODERATESIGNAL4PROBABLYPRESENTLOCALIZED,STRONGSIGNALWITHNODEFINITEMARGINS5DEFINITELYPRESENTLOCALIZED,STRONGSIGNALWITHDEFINITEMARGINSIFALESIONWASCONSIDEREDTOBEPRESENTONAHIGHBVALUEDWMRIMAGE,THELESIONLOCATIONWASRECORDEDONLYLESIONSRECORDEDATTHECORRECTLOCATIONDETERMINEDBYTHESTUDYCOORDINATORSWEREACCEPTEDASTRUEPOSITIVESTATISTICALANALYSISRECEIVEROPERATINGCHARACTERISTICROCCURVESWEREUSEDTOREPRESENTTHEPERFORMANCEOFINDIVIDUALRADIOLOGISTSFORTUMORDETECTIONTHEDIAGNOSTICACCURACYFOREACHRADIOLOGISTWASDETERMINEDBYCALCULATINGTHEAREAUNDERTHEROCCURVEAZGRADES4AND5WEREACCEPTEDASPOSITIVEFORTHEPRESENCEOFCOLORECTALADENOCARCINOMA,ANDTHESENSITIVITYANDSPECIFICITYWERECALCULATEDWITH95CONFIDENCEINTERVALSCISTHEINTEROBSERVERAGREEMENTAMONGREVIEWERSFORTUMORDETECTIONWASCALCULATEDWITHTHELINEARWEIGHTEDKAPPASTATISTICSAKAPPASTATISTICGREATERFIG244YEAROLDMANWITHRECTALADENOCARCINOMAARROWMAXIMUMINTENSITYPROJECTIONMIPRECONSTRUCTEDSAGITTALHIGHBVALUEDIFFUSIONWEIGHTEDMRIMAGEFIG354YEAROLDMANWITHRECTALADENOCARCINOMAARROWSMAXIMUMINTENSITYPROJECTIONMIPRECONSTRUCTEDCORONALHIGHBVALUEDIFFUSIONWEIGHTEDMRIMAGETWOMETASTATICLYMPHNODESAREALSOVISUALIZEDARROWHEADSTHAN075WASCONSIDEREDEXCELLENTAGREEMENTBEYONDCHANCE04–075,FAIRTOGOODAGREEMENTANDLESSTHAN04,POORAGREEMENTRESULTSROCANALYSISYIELDEDAZVALUESOF096,096,AND097FORTHETHREERADIOLOGISTS,RESPECTIVELYTHESENSITIVITIESFORTHETHREERADIOLOGISTSWERE90930/3395CI,747–100,87929/3395CI,709–96,AND90930/3395CI,747–100,RESPECTIVELYFOREACHRADIOLOGIST,THESPECIFICITYWAS10015/1595CI,747–100THEMEANSENSITIVITYANDSPECIFICITYOFHIGHBVALUEDWMRIFORDETECTIONOFCOLORECTALADENOCARCINOMAWERE90930/3395CI,745–976AND10015/1595CI,746–100,RESPECTIVELYFIGURES1,2,AND3SHOWREPRESENTATIVEPATIENTSWITHCOLORECTALADENOCARCINOMASALLKAPPAVALUESINDICATINGINTEROBSERVERAGREEMENTWEREINTHECATEGORYOFEXCELLENTRANGE,083–089DISCUSSIONTOINVESTIGATETHEREASONWHYONLYCOLONICADENOCARCINOMASANDNOTTHEHEALTHYCOLONSHOWEDSTRONGSIGNALINTENSITYONTHEHIGHBVALUEDWMRIMAGESISACHALLENGINGISSUETHATWARRANTSFURTHERSTUDIESHOWEVER,ATHEORETICALEXPLANATIONMIGHTBECONSTRUCTEDBYMEANSOFAWELLDESCRIBEDGENERALHYPOTHESISOFDWMRIITISWELLKNOWNTHATDIFFUSIONISCAUSEDBYRANDOMTRANSLATIONALMOLECULARMOTION,ALSOKNOWNASBROWNIANMOTIONDWMRIISTHEONLYIMAGINGMETHODTHATCANEVALUATETHEDIFFUSIONPROCESSINVIVOTHESPEEDOFDIFFUSIONOFWATERMOLECULESISDIFFERENTINTHEEXTRACELLULARANDINTRACELLULARCOMPONENTSOFTHETISSUES3INTHEINTRACELLULARCOMPONENT,THEDIFFUSIONISRELATIVELYSLOWBECAUSEOFTHEPRESENCEOFCELLULARMEMBRANESTHUS,APPARENTDIFFUSIONCOEFFICIENTSADCS,WHICHAREQUANTITATIVEEXPRESSIONSOFDIFFUSIONCHARACTERISTICSOFTISSUES,ARERELATEDTOTHEPROPORTIONOFEXTRACELLULARANDINTRACELLULARCOMPONENTSTHEYTENDTODECREASEWITHINCREASEDTISSUECELLULARITYORCELLDENSITY8CONVERSELY,THECELLDENSITYMAYBEINDICATIVEOFTUMORAGGRESSIVENESSLYNGETAL9REPORTEDANINCREASEDMETASTATICCAPACITYOFTUMORSWITHHIGHCELLULARITYMOREOVER,INADDITIONTOTHECELLULARMEMBRANES,THEINTRACELLULARCYTOSKELETON,ORGANELLES,MATRIXFIBERS,ANDSOLUBLEMACROMOLECULESCONTRIBUTETODIFFUSIONRESTRICTIONSINTUMORSTHUS,DIFFUSIONCURVESTHATDECAYQUICKLYORLARGEADCVALUESMAYBETYPICALFORHEALTHYTISSUESORBENIGNPATHOLOGICPROCESSESWITHDOWNLOADEDFROMWWWAJRONLINEORGBY59474389ON05/15/13FROMIPADDRESS59474389COPYRIGHTARRSFORPERSONALUSEONLYALLRIGHTSRESERVED

簡介：中文中文5500字出處出處NAKAMURAR,KATAOKAH,SATON,ETALEPHA2/EFNA1EXPRESSIONINHUMANGASTRICCANCERJCANCERSCIENCE,2005,961427EPHA2/EFNA1在人類胃癌中的表達(dá)摘要摘要EPH受體酪氨酸激酶A2EPHA2在人類多種癌癥中過表達(dá)和磷酸纖維化，并伴隨著惡性的轉(zhuǎn)化。然而，最近有一則報(bào)道指出刺激EPHA2配體EFNA1可以抑制胃癌中EPHA2表達(dá)。作者使用半定量PCR方法對胃癌的四個細(xì)胞系和49個原發(fā)的胃癌樣品及其正常胃癌組織的EPHA2和EFNA1表達(dá)進(jìn)行檢測。在27例患者中EPHA2在癌組織中的的表達(dá)比正常組織中高很多（55），而EFNA1在28例患者癌組織中過表達(dá)（57）。在表達(dá)水平和組織學(xué)特征如腫瘤大小、年齡、浸潤或淋巴結(jié)參與之間沒有檢測到顯著的關(guān)系。然而,EPHA2在宏觀3，4型腫瘤中的過表達(dá)比進(jìn)展期胃癌1，2型突出。作者觀察了被檢測的四個胃癌細(xì)胞系中的三個（AGS,KATO3,MKN74）EPHA2的表達(dá)。在一個細(xì)胞系中，TMK1，用NORTHERNBLOTTING，RTPCR，WESTERNBLOTTING幾乎未檢測到EPHA2的表達(dá)。相比，在所有細(xì)胞系中檢測到了EFNA1的表達(dá)。在內(nèi)生表達(dá)EPHA2胃癌細(xì)胞系中由EPHRINA1FC刺激引起EPHA2蛋白表達(dá)下降和提高EPHA2的磷酸化作用。最后，EPHA2的表達(dá)通過可溶的EPHRINA1FC的重復(fù)刺激而抑制。總之，這些表明EPHA2和EFNA1的表達(dá)也許會影響人類胃癌的特征。EPH受體是目前已知的最大的酪氨酸激酶受體家族并可由細(xì)胞表面配體EFN激活。有證據(jù)表明EPH家族中的許多成員及其EFN配體通過細(xì)胞粘附，形成，毛細(xì)管生成參與了血管形成和發(fā)展。EPH受體分成兩類EPHA和EPHB。EPHA的相對應(yīng)的配體是EFNA,EPHB的是EFNB配體。EPH的表達(dá)轉(zhuǎn)錄已在黑素瘤和癌癥中證實(shí)。EPHA2的過表達(dá)被認(rèn)為在乳房上皮細(xì)胞轉(zhuǎn)化為惡性時很重要。EFNA2過表達(dá)食管鱗狀細(xì)胞癌預(yù)后比EFNA2表達(dá)低的差。胃癌預(yù)后很差，激酶在胃癌細(xì)胞中的作用是研究的重點(diǎn)。OGAWA等在2000年鑒定在一些胃癌病例中EFNA1和EPHA2有表達(dá)，但是他們的分子作用還不清楚，盡管在胃癌中對酪氨酸激酶進(jìn)一步研究。所以，作者用定量PCR，RTPCR,NORTHERN雜交，免疫印跡雜交對胃癌樣本和胃癌細(xì)胞系的EPHA2和EFNA1的表達(dá)水平進(jìn)行了檢測。這是首例報(bào)道EFNA1和EPHA2在胃癌中有表??‘N1,N2ANDN3’用的也是JCS法T檢驗(yàn)法和秩和檢驗(yàn)法。細(xì)胞培養(yǎng)細(xì)胞培養(yǎng)在添加10胎牛血清的RPMI1640中培養(yǎng)胃癌細(xì)胞系（KATO3,MKN74,TMK1），而AGS細(xì)胞系用添加10胎牛血清的HAM’SF12K培養(yǎng)基培養(yǎng)。293T人類腎胚胎細(xì)胞系用已添加10胎牛血清的DMEM培養(yǎng)基培養(yǎng)。RNA提取和反轉(zhuǎn)錄提取和反轉(zhuǎn)錄根據(jù)廠家紀(jì)錄使用ISOGENRNA提取試劑盒從人類組織中提取總RNA。從總的RNA得到單鏈CDNA，20L體系中還含有1GOLIGODT隨機(jī)引物，MMULVRT反轉(zhuǎn)錄酶，RNA酶抑制劑。定量定量PCR分析分析本研究中使用了比先前方法改進(jìn)的定量PCR方法。簡要說，CDNA用水稀釋，20?L體系中混0625?MOL/L引物，1UTAQDNA酶（羅氏）和1CIDCTP（AMERSHAMBIOSCIENCES,PISCATAWAY,NJ,USA，使用DNA循環(huán)器擴(kuò)增（PC700ASTEC,FUKUOKA,JAPAN。對照??ACTIN，30個循環(huán)，包括94℃變性45S,59℃退火1MIN,72℃1MIN,最后延伸72℃10MIN。EPHA2ANDEFNA1使用35個循環(huán)94℃變性45S,59℃退火1MIN,72℃1MIN,最后延伸72℃10MIN。用這些條件對ACTB,EPHA2,EFNA1進(jìn)行指數(shù)冪擴(kuò)增。每個反應(yīng)設(shè)立陰性對照排除DNA的污染。通過ACTBMRNA在相同樣品存在及在28S,18S中的峰（使用AGILENT2100BIOANALYZER）來證實(shí)從臨床標(biāo)本提取RNA的完整性。ACTB,EPHA2,EFNA1的片段大小分別為121BP,260BP,230P。引物序列為（A（EPHA2上游5′GCAACATCCTCGTCAACAGC3′下游5′TGGCTTTCATCACCTCGTGG3′（B（?FHA1上游5′AACAAGCTGTGCAGGCATGG3′下游5′CTCCACAGATGAGGTCTTGC3′（C（ACTB上游5′GCTACGTCGCCCTGGACTTC3′下游5′AGCGGAACCGCTCATTGCCA3′PCR產(chǎn)物用6的瓊脂糖電泳檢測，然后用凝膠成像分析儀（MACBAS分析。在胃癌和對應(yīng)的非腫瘤組織中具有代表性的EFNA1,EPHA2和ACTB的表達(dá)圖件圖1

簡介：中文中文3750字，字，2400單詞，單詞，13萬英文字符萬英文字符出處出處ELSAYEDZM,ABOUELHSCLINICALEVALUATIONOFGENPROBESAMPLIFIEDMYCOBACTERIUMTUBERCULOSISDIRECTTESTFORRAPIDDIAGNOSISOFMYCOBACTERIUMTUBERCULOSISINEGYPTIANCHILDRENATRISKFORINFECTIONJARCHIVESOFPATHOLOGYSAMIRABOUELHASSAN摘要摘要內(nèi)容結(jié)核?。═B）的診斷檢測實(shí)驗(yàn)已大為改善。樣品處理后4至7小時內(nèi)，我們就可以得到可靠的合理的，標(biāo)準(zhǔn)化的，基于核酸的擴(kuò)增技術(shù)的檢測結(jié)果，目的研究抗酸涂片檢查陽性或陰性的埃及高結(jié)核感染風(fēng)險(xiǎn)兒童的BACTEC460TB培養(yǎng)比較基因芯片法檢測結(jié)核分枝桿菌的診斷效能。設(shè)計(jì)前瞻性評估了50個陽性結(jié)核病史的家庭孩子。所有患者均來自埃及曼蘇拉大學(xué)兒童醫(yī)院門診。孩子們PPD試驗(yàn)硬結(jié)直徑超過10毫米，并在過去兩年內(nèi)接種卡介苗疫苗疤痕。每個病人連續(xù)取三個痰標(biāo)本。對樣品進(jìn)行了基因芯片法檢查，直接涂片鏡檢，細(xì)菌培養(yǎng)BACTEC460TB。結(jié)果50例中，有30（60％）的痰標(biāo)本呈BACTEC460TB陽性反應(yīng)，29例（58％）基因芯片法呈陽性?？顾嵬科瑱z查的敏感性和特異性分別為833％和100％，準(zhǔn)確度均為90％。基因芯片法的敏感度和特異度分別為967％和100％，準(zhǔn)確度98％。結(jié)論本研究結(jié)果表明，基因芯片法檢測兒童結(jié)核病呼吸道樣本中的結(jié)核分枝桿菌復(fù)合物是一種快速、精確的方法，它可以用來診斷涂片陰性的疑似結(jié)核病人。前言前言兒童結(jié)核病（TB）在與成人結(jié)核患者接觸的兒童中發(fā)病率最高。本研究的動機(jī)是觀察到兒童結(jié)核病例的數(shù)量正在增加。傳統(tǒng)分枝桿菌培養(yǎng)的實(shí)驗(yàn)室診斷通常需要2至8周。近期增加新的結(jié)核病例表明，急需新的快速，特異的診斷檢測結(jié)核分枝桿菌的方法。隨著新的分子生物學(xué)技術(shù)的發(fā)展，相信不就就會到來。核酸擴(kuò)增檢測是快速的和明確的。盡管理論上能夠探測到甚至一個單一的分枝桿菌細(xì)胞，但不知核酸擴(kuò)增試驗(yàn)（核酸擴(kuò)增試驗(yàn)）是否足夠可靠到可以取代傳統(tǒng)的診斷方法檢測結(jié)核。此試驗(yàn)的特點(diǎn)和測試程序中的錯誤可能造成不準(zhǔn)確。此外，在呼吸道分泌物的酶，能夠抑制擴(kuò)增反應(yīng)造成額外的3％至25％的假陰性結(jié)果的存在。另一方面，出現(xiàn)假陽性結(jié)果通常由于陰性樣品的污染與任何生物或靶DNA從含有大量分枝桿菌的樣品或從擴(kuò)增子在實(shí)驗(yàn)室房間污染。為了克服這些問題，開發(fā)出更強(qiáng)大的標(biāo)準(zhǔn)化程序和樣品處理，擴(kuò)增及檢測試劑使用自動化商業(yè)系統(tǒng)。各種自動化系統(tǒng)，擴(kuò)增和檢測技術(shù)的基礎(chǔ)上，已經(jīng)設(shè)計(jì)出用于檢測臨床樣品中的結(jié)核分枝桿菌。該系統(tǒng)包括基于聚合酶鏈反應(yīng)COBASAMPLICOR系統(tǒng)分枝桿菌（羅氏診斷，巴塞爾，瑞士）轉(zhuǎn)錄介導(dǎo)的，基于擴(kuò)增，擴(kuò)增結(jié)核分枝桿菌直接試驗(yàn)（AMTDGENPROBE公司，公司，圣地亞哥，加州）的8鏈置換擴(kuò)增基于BDPROBETEC的ET系統(tǒng)（碧迪公司，新澤西州富蘭克林湖）9和連接酶鏈反應(yīng)為基礎(chǔ)的雅培LCX結(jié)核分枝桿菌檢測系統(tǒng)（雅培，北芝加哥，Ⅲ）。由GENPROBE公司AMTD檢測通過使用特定的RNA轉(zhuǎn)錄介導(dǎo)的等溫?cái)U(kuò)增方法。這項(xiàng)工作的目的是研究基因芯片技術(shù)直接檢測結(jié)核分枝桿菌比較BACTEC460TB培養(yǎng)檢查對抗酸（ZN）涂片陰性或陽性埃及高結(jié)核感染風(fēng)險(xiǎn)兒童的診斷性能。材料與方法材料與方法病人的選擇病人的選擇本研究前瞻性評估了50個孩子，介于10至15歲的陽性結(jié)核病史的家庭。所有患者均來自埃及曼蘇拉大學(xué)兒童醫(yī)院門診。孩子們具有咳嗽癥狀。他們PPD陽性，結(jié)果為直徑超過10毫米的硬結(jié)，并在過去2年接種過卡介苗（BCG）。獲得所有參與者的父母知情的書面同意，曼蘇拉大學(xué)醫(yī)院倫理委員會批準(zhǔn)了這項(xiàng)研究。在美國17報(bào)道TB幼兒中心爆發(fā)，因?yàn)樵趫龅某赡耆嘶加薪Y(jié)核病。與家人密切的互動成為延誤診斷結(jié)核病傳染給孩子的主要途徑。小兒結(jié)核病的增加需要我們提高臨床診斷能力。改善結(jié)核病控制和針對性的宣傳計(jì)劃，在埃及高風(fēng)險(xiǎn)的移民人口可能會增加兒童和成人的病例，需要一個有效的檢測方法來診斷和治療結(jié)核病。在本研究中一個有趣的發(fā)現(xiàn)是，所有陽性X射線檢查結(jié)果的患者的也有陽性的培養(yǎng)。然而，X射線發(fā)現(xiàn)異常的患者中只有40％。同樣，SWINGLER等19報(bào)道，在兒童結(jié)核病的診斷與檢測縱隔淋巴結(jié)腫大的準(zhǔn)確性是值得商榷的。因此，我們必須選擇更精確的方法，以免誤診那些病人。當(dāng)評估ZN涂抹作為一個快速和廉價(jià)的結(jié)核病診斷方法的性能，特異度為100％，敏感度為833％，90％的整體精度。雖然結(jié)核分枝桿菌培養(yǎng)被認(rèn)為是黃金標(biāo)準(zhǔn)檢測方法，但顯微鏡足夠迅速，在24小時內(nèi)提供結(jié)果，可是，后者缺乏敏感性且無法分辨其他分枝桿菌結(jié)核菌?；蛐酒ㄅc培養(yǎng)結(jié)果相比，29例基因芯片法培養(yǎng)陽性，1例基因芯片法陰性培養(yǎng)陽性。4個樣品培養(yǎng)和基因芯片法及AFB涂片均呈陽性的。一般來說，陽性和陰性對照和標(biāo)本的臨界值之間的差異是不夠明顯。這解釋了培養(yǎng)陽性標(biāo)本的基因芯片法結(jié)果陰性，可能因?yàn)樯贁?shù)分枝桿菌的分布不均，很顯然，基因芯片法技術(shù)增加了診斷結(jié)核病的百分比。格雷科等23報(bào)道了類似的結(jié)論，對于大多數(shù)的自動化系統(tǒng)。根據(jù)AFB涂片的結(jié)果，患者預(yù)后的影響會有所不同。在涂片陽性的患者，主要是影響公眾健康和醫(yī)院感染控制資源措施。AFB涂片陰性時，影響患者預(yù)后的潛力更大。在涂片陰性的患者，基因芯片法可以提供更快速診斷結(jié)核病和隨后開始治療，這避免了需要侵入性的診斷方法，并允許住院患者提前出院，這些侵入性診斷方法是昂貴的，會對病人構(gòu)成風(fēng)險(xiǎn)。因此，在發(fā)展中國家，如埃及，我們可以對疑似病例開展基因芯片法檢查，降低診斷費(fèi)用。培養(yǎng)與涂片檢查的成本是25元左右，基因芯片法每箱成本為50美元左右。我們得出的數(shù)據(jù)，顯示了基因芯片法出色的靈敏度和特異度。基因芯片法測試的靈敏度為967％，特異度為100％，陽性預(yù)測值為100％，陰性預(yù)測值為952％，準(zhǔn)確度為98％。涂片陰性樣品敏感性為80％。王和泰有類似的報(bào)道，基因芯片法的靈敏度為984％，特異度為100％，陽性預(yù)測值為100％。在本研究中，基因芯片法結(jié)果證明，對于結(jié)核高風(fēng)險(xiǎn)兒童，作為一種快速，準(zhǔn)確診斷肺結(jié)核的方法是有價(jià)值的。其結(jié)果可在4小時內(nèi)得到，而培養(yǎng)結(jié)果可能在14天內(nèi)獲得。我們可以不依賴于ZN單獨(dú)作為一個快速的方法，因?yàn)樗撵`敏度很低。在埃及的情況下，控制結(jié)核病接觸有許多困難。雖然官方程序推薦所有接觸結(jié)核病患者進(jìn)行篩選，診斷策略的缺乏阻止了大量的結(jié)核病病例的早期診斷，從而提高了疾病的傳播。我們可以建議使用簡單的測試，如胸部X射線和ZN染色，兒童結(jié)核病的風(fēng)險(xiǎn)組合。對于那些具有陰性結(jié)果患者，我們可以根據(jù)情況進(jìn)行培養(yǎng)檢查或基因芯片法檢查。這項(xiàng)研究的結(jié)果表明，基因芯片法是一種精確的方法，可以快速檢測結(jié)核分枝桿菌復(fù)合物在兒童疑似結(jié)核病例的呼吸道樣本，特別是檢測涂片陰性結(jié)核疑似病例。

簡介：REVIEWARTICLEREVIEWTHEROLEOFMICRORNASINKIDNEYDISEASENEP_1363599608JORDANYZLI,1,2TUCKYYONG,2,3MICHAELZMICHAEL2,4ANDJONATHANMGLEADLE1,2DEPARTMENTSOF1RENALMEDICINEAND3GENERALMEDICINEAND4GASTROENTEROLOGYANDHEPATOLOGY,FLINDERSMEDICALCENTRE,AND2SCHOOLOFMEDICINE,FLINDERSUNIVERSITY,ADELAIDE,SOUTHAUSTRALIA,AUSTRALIAKEYWORDSBIOMARKER,DIABETICNEPHROPATHY,EPITHELIALMESENCHYMALTRANSITION,KIDNEYDISEASE,MICRORNACORRESPONDENCEPROFESSORJONATHANGLEADLE,DEPARTMENTOFRENALMEDICINE,LEVEL6,FLINDERSMEDICALCENTRE,FLINDERSDRIVE,BEDFORDPARK,SA5042,AUSTRALIAEMAILJONATHANGLEADLEHEALTHSAGOVAUACCEPTEDFORPUBLICATION30MAY2010ACCEPTEDMANUSCRIPTONLINE7JUNE2010DOI101111/J14401797201001363XSUMMARYATAGLANCETHISISACOMPREHENSIVEANDSCHOLARLYREVIEWOFTHECURRENTKNOWLEDGEOFMICRORNASMIRNASINRENALDISEASEMIRNASAREEMERGINGASIMPORTANTREGULATORSOFDISEASEPROCESSESUNDERSTANDINGHOWMIRNASMODULATEPATHOGENETICPATHWAYSISIMPORTANT,ASTHERAPEUTICMANIPULATIONOFMIRNASMAYEVOLVEASAPOTENTIALSTRATEGYFORTREATINGRENALDISEASESINTHEFUTUREABSTRACTMICRORNASMIRNASARESHORTNONCODINGRNASTHATMODULATEPHYSIOLOGICALANDPATHOLOGICALPROCESSESBYINHIBITINGTARGETGENEEXPRESSIONVIABLOCKADEOFPROTEINTRANSLATIONORBYINDUCINGMRNADEGRADATIONTHESEMIRNASPOTENTIALLYREGULATETHEEXPRESSIONOFTHOUSANDSOFPROTEINSASARESULT,MIRNASHAVEEMERGEDRAPIDLYASAMAJORNEWAREAOFBIOMEDICALRESEARCHWITHRELEVANCETOKIDNEYDISEASEMIRNAEXPRESSIONHASBEENSHOWNTODIFFERBETWEENTHEKIDNEYANDOTHERORGANSASWELLASBETWEENDIFFERENTKIDNEYREGIONSFURTHERMORE,MIRNASHAVEBEENFOUNDTOBEFUNCTIONALLYIMPORTANTINMODELSOFPODOCYTEDEVELOPMENT,DIABETICNEPHROPATHYANDPOLYCYSTICKIDNEYDISEASEOFPARTICULARINTEREST,PODOCYTESPECIFICDELETIONOFDICER,AKEYENZYMEINTHEBIOGENESISOFMIRNA,RESULTSINPROTEINURIAANDSEVERERENALIMPAIRMENTINMICEONEMIRNAMIR192CANALSOACTASANEFFECTOROFTRANSFORMINGGROWTHFACTORBACTIVITYINTHEHIGHGLUCOSEENVIRONMENTOFDIABETICNEPHROPATHYDIFFERENTIALEXPRESSIONOFMIRNASHASBEENREPORTEDINKIDNEYALLOGRAFTREJECTIONITISANTICIPATEDTHATFUTURESTUDIESINVOLVINGMIRNASWILLGENERATENEWINSIGHTSINTOTHECOMPLEXPATHOPHYSIOLOGYUNDERLYINGVARIOUSKIDNEYDISEASES,GENERATEDIAGNOSTICBIOMARKERSANDMIGHTBEOFVALUEASTHERAPEUTICTARGETSFORPROGRESSIVEKIDNEYDISEASESTHEPURPOSEOFTHISREVIEWISTOHIGHLIGHTKEYMIRNADEVELOPMENTSINKIDNEYDISEASESANDHOWTHISMIGHTINFLUENCETHEDIAGNOSISANDMANAGEMENTOFPATIENTSWITHKIDNEYDISEASEINTHEFUTUREINTRODUCTIONMICRORNASMIRNASAREENDOGENOUSNONCODINGRNAMOLECULES,20–22NUCLEOTIDESINLENGTHTHEDISCOVERYANDCHARACTERIZATIONOFMIRNAINTHELASTDECADEISREVOLUTIONIZINGOURUNDERSTANDINGOFGENEREGULATION,CELLDIFFERENTIATION,PROLIFERATION,APOPTOSIS,METABOLISMANDPATHOPHYSIOLOGYOFMANYDISEASESINCLUDINGKIDNEYDISEASESTHEUNDERSTANDINGOFMIRNABIOLOGYANDITSROLEINVARIOUSDISEASESISSTILLINITSEARLYSTAGEBUTISEXPANDINGRAPIDLYTHEAIMOFTHISREVIEWISTOHIGHLIGHTMIRNABIOLOGYINRELATIONTOKIDNEYDISEASESANDITSIMPORTANCEINUNDERSTANDINGDISEASEMECHANISMSFUTUREDIRECTIONSINTHISFIELDWILLALSOBEDISCUSSEDDISCOVERYANDBIOGENESISOFMICRORNASMIRNASWEREFIRSTFOUNDINTHENEMATODECAENORHABDITISELEGANSIN19931SINCETHENTHEYHAVEALSOBEENDESCRIBEDWIDELYINPLANTSANDMAMMALS2MIRNASAREFIRSTTRANSCRIBEDINTHENUCLEUSASSTEMLOOPPRIMARYMIRNA,WHICHARETHENCLEAVEDINTOSHORTERPRECURSORMIRNABYDROSHA,ANRNASEIII,ANDITSESSENTIALCOFACTORCALLEDDGCR8DIGEORGESYNDROMECRITICALREGION8,ADOUBLESTRANDEDRNABINDINGPROTEINFIG13–6THEPRECURSORMIRNASARETRANSPORTEDOUTOFTHENUCLEUSVIAEXPORTIN5ANDONCEINTHECYTOSOLARECLEAVEDINTOTHEIRMATUREFORMOF20–22NUCLEOTIDESBYDICER,ANOTHERRNASEIII7,8AFTERCLEAVAGE,THEMIRNADUPLEXISUNWOUNDANDTHEFUNCTIONALSTRANDISLOADEDONTOTHERNAINDUCEDSILENCINGCOMPLEXRISCANDFUNCTIONSASITSGUIDE9THEMATUREMIRNAGUIDESTHERISCCOMPLEXTOANEARCOMPLEMENTARYSEQUENCE,USUALLYINTHE3′UNTRANSLATEDREGIONUTR,OFATARGETMESSENGERRNAMRNA9UPONBINDING,THERISCCAUSESPOSTTRANSCRIPTIONALGENESILENCINGBYEITHERCLEAVINGTHETARGETMRNAORBYINHIBITINGITSTRANSLATION,SOTHATMIRNASAREUSUALLYNEGATIVEREGULATORSOFGENEEXPRESSION10INADDITIONTOTHEIRROLEINSUCHPOSTTRANSCRIPTIONALREPRESSION,MIRNASHAVENOWBEENIMPLICATEDNEPHROLOGY152010599–608?2010THEAUTHORSNEPHROLOGY?2010ASIANPACIFICSOCIETYOFNEPHROLOGY599ENABLEMONITORINGINACLINICALSETTINGORIGINALLY,RNABLOTANALYSESPROVIDEDBOTHQUANTITATIVEANDQUALITATIVEINFORMATIONABOUTTHEVARIOUSFORMSOFAMIRNAWITHINATOTALRNASAMPLE1,16ASTHENUMBEROFMIRNASINTHEMIRBASEREGISTRY17HASINCREASED,MICROARRAYTECHNOLOGYHASBEENADAPTEDTOENABLETHEPARALLELSCREENINGOFTHOUSANDSOFMIRNASINONESAMPLE18MORERECENTLY,REALTIMEREVERSETRANSCRIPTIONPOLYMERASECHAINREACTIONHASBEENADAPTEDTOENABLERELATIVEQUANTIFICATIONANDQUANTITATIVEANALYSISOFMIRNALEVELSSPECIFICAMPLIFICATIONOFMATUREMIRNASCANBEACHIEVEDUSINGSTEMLOOPOLIGONUCLEOTIDESTOPRIMEREVERSETRANSCRIPTIONANDENABLETAQMANDETECTION,19WHILEALTERNATEREAGENTSANDMETHODSFORAMPLIFYINGPRECURSORANDPRIMARYTRANSCRIPTSAREALSOCOMMERCIALLYAVAILABLEMICROFLUIDICSYSTEMSNOWENABLEHIGHTHROUGHPUTMIRNAPCRPROFILINGWITHSMALLAMOUNTSOFINPUTSAMPLERNA,ENABLINGANALYSISOFSMALLBIOPSIES,LIMITEDVOLUMESOFBODYFLUIDS,OREVENFORMALINFIXEDPARAFFINEMBEDDEDARCHIVALMATERIAL20THEHYBRIDIZATIONKINETICSOFOLIGONUCLEOTIDESHAVEBEENENHANCEDTHROUGHTHEINCORPORATIONOFLOCKEDNUCLEICACIDMONOMERS,WHICHPROVIDEANADVANTAGEFORPCRANDINSITUHYBRIDIZATION21ANDALSOENHANCETHEPOTENTIALFOREMPLOYINGANTIMIRNASTRATEGIESINTHERAPEUTICROLES22,23MICRORNASINRENALPHYSIOLOGYTHESUGGESTIONOFORGANSPECIFICROLESFORMIRNASEMERGEDWITHTHEDEMONSTRATIONOFTISSUERESTRICTEDMIRNAEXPRESSION,INCLUDINGCLUSTERSOFMIRNASTHATAREEXPRESSEDSPECIFICALLYINTHEKIDNEY24CONVERSELY,THEABSENCEORLOWERLEVELSOFPARTICULARMIRNASINTHEKIDNEYCOMPAREDWITHOTHERORGANSMAYPERMITRENALSPECIFICEXPRESSIONOFTARGETPROTEINSTHATAREIMPORTANTFORKIDNEYFUNCTION24,25EXAMPLESOFMIRNASTHATAREMOREABUNDANTINTHEKIDNEYCOMPAREDWITHOTHERORGANSINCLUDEMIR192,MIR194,MIR204,MIR215ANDMIR216TIANETALESTABLISHEDTHEFIRSTDIFFERENTIALPROFILEOFMIRNAEXPRESSIONBETWEENTHERENALCORTEXANDMEDULLAOFRATSINDICATINGAPOTENTIALROLEINTISSUESPECIFICATION26HOWEVER,CELLTYPESPECIFICMIRNASINTHEKIDNEYHAVENOTYETBEENREPORTEDACRITICALROLEOFMIRNAREGULATIONINTHEPROGRESSIONOFGLOMERULARANDTUBULARDAMAGE,ANDTHEDEVELOPMENTOFPROTEINURIAHAVEBEENSUGGESTEDBYSTUDIESINMICEWITHPODOCYTESPECIFICDELETIONOFDICER27–29ALLTHREEREPORTSSHOWEDMAJORRENALABNORMALITIESINTHESEMICEINCLUDINGPROTEINURIA,PODOCYTEFOOTPROCESSEFFACEMENT,GLOMERULARBASEMENTMEMBRANEABNORMALITIES,PODOCYTEAPOPTOSIS,PODOCYTEDEPLETIONANDMESANGIALEXPANSIONTHEREWASARAPIDPROGRESSIONOFRENALDISEASEWITHINITIALDEVELOPMENTOFALBUMINURIAFOLLOWEDBYPATHOLOGICALFEATURESOFGLOMERULOSCLEROSISANDTUBULOINTERSTITIALFIBROSISTHISLEDTORENALFAILUREANDDEATHBY6–8WEEKSITISLIKELYTHATTHESEPHENOTYPESAREDUETOTHEGLOBALLOSSOFMIRNASBECAUSEOFDICERDELETION,BUTGIVENMULTIPLEMIRNASANDTHEIRMYRIADTARGETS,THEPRECISEPATHWAYSRESPONSIBLEREQUIREIDENTIFICATIONTHESEINVESTIGATORSALSOIDENTIFIEDSPECIFICMIRNACHANGES,FOREXAMPLE,THEDOWNREGULATIONOFTHEMIR30FAMILYWHENDICERWASDELETEDOFRELEVANCE,THEMIR30FAMILYWASFOUNDTOTARGETCONNECTIVETISSUEGROWTHFACTOR,APROFIBROTICMOLECULETHATISALSODOWNSTREAMOFTRANSFORMINGGROWTHFACTORTGFB30THUS,THETARGETSOFTHESEMIRNASMAYREGULATECRITICALGLOMERULARANDPODOCYTEFUNCTIONSTHESEFINDINGSHAVEALSOBEENCOMPLEMENTEDBYANELEGANTSTUDYREVEALINGADEVELOPMENTALROLEFORTHEMIR30FAMILYDURINGPRONEPHRICKIDNEYDEVELOPMENTINXENOPUS31RECENTLY,ANOTHERSTUDYHASSHOWNTHATDELETIONOFDICERINTHERENINSECRETINGCELLSOFMICESEVERELYREDUCEDTHENUMBEROFJUXTAGLOMERULARCELLS,DECREASEDEXPRESSIONOFTHERENINGENES,LOWEREDPLASMARENINCONCENTRATIONANDDECREASEDBLOODPRESSURE32THEKIDNEYSDEVELOPEDSTRIKINGVASCULARABNORMALITIESANDPROMINENTSTRIPEDFIBROSISTHESEFINDINGSHIGHLIGHTTHEIMPORTANTROLESOFDICERANDMIRNASINRENALPHYSIOLOGYANDPATHOLOGY,ALTHOUGHTHEEXTENTTOWHICHSUCHGENETICSTUDIESREVEALANESSENTIALANDFUNDAMENTALROLEOFDICERINCELLULARFUNCTION,ASOPPOSEDTOASPECIFICROLEINRENINSECRETINGCELLS,ISARGUABLETHEIMPORTANCEOFDICERINCELLULARFUNCTIONISFURTHERHIGHLIGHTEDBYWEI’SSTUDY33THEYESTABLISHEDAMOUSEMODELWITHTARGETEDDICERDELETIONINRENALPROXIMALTUBULESTHESEMICEHADNORMALRENALFUNCTIONANDHISTOLOGYDESPITEAGLOBALDOWNREGULATIONOFMIRNASINTHERENALCORTEXHOWEVER,THESEMICEWERESTRIKINGLYRESISTANTTORENALISCHAEMIAREPERFUSIONINJURY,SHOWINGSIGNIFICANTLYBETTERRENALFUNCTION,LESSTISSUEDAMAGE,LOWERTUBULARAPOPTOSISANDIMPROVEDSURVIVALCOMPAREDWITHTHEIRWILDTYPECOUNTERPARTS33MICRORNASINRENALDISEASESDIABETICNEPHROPATHYDIABETICNEPHROPATHYISTHELEADINGCAUSEOFENDSTAGEKIDNEYDISEASEBUTOURUNDERSTANDINGOFTHEDISEASEMECHANISMSISINCOMPLETESTUDIESOFMIRNAEXPRESSIONINDIABETICNEPHROPATHYHAVESOFAREMERGEDPREDOMINANTLYFROMANIMALMODELSOFDIABETESANDTHEEFFECTSOFHYPERGLYCAEMIAINONESTUDY,MIR192LEVELSWERESHOWNTOBEINCREASEDINGLOMERULIISOLATEDFROMSTREPTOZOTOCININJECTEDDIABETICMICEANDDIABETICMICEDB/DBWHENCOMPAREDWITHNONDIABETICMICE34INTHISSTUDY,MIR192WASSHOWNTOREGULATEEBOXREPRESSORSTHATARERESPONSIBLEFORCONTROLLINGTHEEXPRESSIONOFTGFBINDUCEDEXTRACELLULARMATRIXPROTEINS,COLLAGEN1A1AND2COL1A1AND2COL1A1AND2WERESHOWNTOACCUMULATEDURINGDIABETICNEPHROPATHYTHEREFORE,THESERESULTSSUGGESTAPOTENTIALROLEOFMIR192INDIABETICNEPHROPATHYORTHATMIR192CANBEANEFFECTOROFTGFBHOWEVER,DISCORDANTLYARECENTSTUDYDEMONSTRATEDTHATMIR192EXPRESSIONISDECREASEDINPROXIMALTUBULAREPITHELIALCELLSINRESPONSETOTGFB35THELOSSOFMIR192CORRELATESWITHTUBULOINTERSTITIALFIBROSISANDREDUCTIONINEGFRINRENALBIOPSIESFROMPATIENTSWITHESTABLISHEDDIABETICNEPHROPATHYMICRORNASINKIDNEYDISEASE?2010THEAUTHORSNEPHROLOGY?2010ASIANPACIFICSOCIETYOFNEPHROLOGY601

簡介：1中文中文3890字出處出處TANIèREP,MARTELPLANCHEG,MAURICID,ETALMOLECULARANDCLINICALDIFFERENCESBETWEENADENOCARCINOMASOFTHEESOPHAGUSANDOFTHEGASTRICCARDIAJAMERICANJOURNALOFPATHOLOGY,2001,15813340賁門癌和食管癌在分子和臨床上的不同賁門癌和食管癌在分子和臨床上的不同摘要帶有BARRETT’S粘膜的食管腺癌（ADCE）和賁門腺癌（ADCC）經(jīng)常被認(rèn)為是同一種病理疾病。在本研究中，我們用嚴(yán)格的解剖病理標(biāo)準(zhǔn)來區(qū)分這兩種損傷，我們發(fā)現(xiàn)兩者在TP53突變，MDM2基因擴(kuò)增和細(xì)胞角蛋白表達(dá)上有著不同。從26個ADCC和28個ADCE患者的福爾馬林固定，石蠟包埋的切片中提取了DNA。TP53突變用暫時溫度梯度電泳檢測，用測序鑒定。MDM2基因擴(kuò)增用DIFFERENTIALPCR評定。細(xì)胞角蛋白4，7，13的表達(dá)用免疫組化技術(shù)檢測。在ADCC，男女比例為181，在ADCE中為271。5個ADCC患者有其他腫瘤的病史，在ADCE中有一個。兩種腫瘤在TP53突變31在ADCC中，50在ADCE中，MDM2基因擴(kuò)增19在ADCC中，4在ADCE中,細(xì)胞角蛋白7在ADCE中100陽性，在ADCC中41陽性和細(xì)胞角蛋白13（在ADCE中81陽性，在ADCC中365陽性）表達(dá)方式上有著不同。ADCE和ADCC在臨床特點(diǎn)，在TP53突變，MDM2基因擴(kuò)增和細(xì)胞角蛋白表達(dá)上有著不同。這些結(jié)果支持ADCE和ADCC是不同的病理疾病的說法。背景介紹過去的二十年中，美國和一些西歐國家胃食管接合部腫瘤的發(fā)病率以每年510的速率在增加1。增加的原因還不清楚。胃食管接合部腫瘤主要有兩種食管腺癌ADCE和賁門腺癌ADCC。ADCE出現(xiàn)在食管遠(yuǎn)端，由BARRETT’S粘膜發(fā)展而來。BARRETT’S粘膜是鱗狀上皮的腺樣化生，高度在幾毫米到幾厘米之間。有證據(jù)表明，化生的粘膜細(xì)胞是雜合細(xì)胞，既表達(dá)鱗狀細(xì)胞來源的細(xì)胞角蛋白CK4AND13，又表達(dá)腺細(xì)胞來源的細(xì)胞角蛋白CK8AND192，還具有鱗狀細(xì)胞和腺細(xì)胞的超微結(jié)構(gòu)的特征3。而且，還常常表達(dá)CK7，而在賁門粘膜的腸上皮化生的細(xì)胞中從不表達(dá)4。BARRETT’S粘膜常常與慢性胃食管返流有關(guān)。它也伴隨慢性膽道堿性返流的出現(xiàn)5。易引起B(yǎng)ARRETT’S粘膜的因素還未完全理清。近來的證據(jù)表明，谷胱甘肽S轉(zhuǎn)移酶P1的某種多態(tài)表達(dá)可能是發(fā)展為BARRETT’S粘膜的遺傳易感性的因素6。在美國的一般人群中，BARRETT’S粘膜是很常見的損傷（10）。賁門是食管和胃之間的移行部位。用肉眼不能鑒別。在顯微鏡下，賁門是一層有少量腺細(xì)胞的薄粘膜，而無泌酸細(xì)胞。高度在1MM到5MM之間，隨著年齡的增長而增加。“ADCC“的意思是位于賁門區(qū)域的腫瘤。但并未明確腫瘤是來源于真正的賁門粘膜還是附近的上基底粘膜。賁門處可有腸上皮化生，尤其在有慢性炎癥的時候。但還沒有明確的證據(jù)表明化生能發(fā)展為ADCC。在許多研究中，常把ADCC和ADCE合稱為胃食管接合部腺癌，但這樣就會排除了返流以外的特殊的危險(xiǎn)因素。吸煙的作用還在爭論811。肥胖癥和抗高血壓藥能松弛賁門括約肌，有助于慢性胃食管返流，故被認(rèn)為是間接的危險(xiǎn)因素。攝取高熱量和脂肪也有可能12，13。相反，攝取較多的纖維，煙酸，維生素B6，鐵和鋅則有助于防止腫瘤的發(fā)展8，10，12。在分子水平上，TP53突變是胃食管接合部腫瘤中最常見的改變。在ADCE中，58的病例有TP53突變。最常見的類型是CPG上的C向T突變50。突變在腫瘤形成早期就已出3梯度電泳。陰性對照（野生型）和陽性對照（已知突變）也用上述分析。顯示有額外和/或異常帶的樣品再用基因組DNA和暫時溫度梯度電泳檢測。如果確定了，從第二個凝膠中切下突變的等位基因，用相同的引物再擴(kuò)增一次，用不對襯PCR擴(kuò)增后，直接測序分析20,22。有兩個P53免疫染色陽性的樣本，在暫時溫度梯度電泳中未顯示有異常帶的可再生模型（1樣本，表1；2樣本，表2）。在這兩個樣本中，MRNA從冰凍的活檢標(biāo)本中提取，CDNA用FLAMAN等人的23酵母菌功能測定法準(zhǔn)備和測定。陽性克隆用ABIPRISM310基因分析機(jī)PERKINELMERBIOSYSTEMS,FOSTERCITY,CA測序。MDM2基因擴(kuò)增的分析用改進(jìn)的差異PCR5ΜL的模板DNA在含有20PMOL對MDM2和多巴胺D2受體基因DRD2引物，200ΜMOL/L每個DNTP,1X擴(kuò)增緩沖液,1XQ液體和05ΜL25UOFHOTSTARTAQDNA聚合酶QIAGEN的50ΜL反應(yīng)混合物。PCR的條件是95°C15分鐘，95°C45秒27個循環(huán)，55°C45秒，72°C1分鐘，72°C擴(kuò)增5分鐘后結(jié)束。引物如下5GAGGGCTTTGATGTTCCTGA3SENSE和5GCTACTAGAAGTTGATGGC3ANTISENSE對MDM2,和5CCACTGAATCTGTCCTGGTATG3SENSE和5GTGTGGCATAGTAGTTGTAGTGG3ANTISENSE對人DRD2。PCR產(chǎn)物用75聚丙酰胺凝膠電泳，溴乙錠染色，照相，膠卷中掃描密度測定法分析GS670BIORAD,HERCULES,CA。MDM2/DRD2的比值在25或25以上，則認(rèn)為MDM2又?jǐn)U增，若比值在2到25之間則認(rèn)為可能有MDM2擴(kuò)增。統(tǒng)計(jì)學(xué)評價(jià)自變量的頻率表用PEARSON’S卡方檢驗(yàn)來統(tǒng)計(jì)。結(jié)果病人的臨床和個體特點(diǎn)19951999年之間收集了26個ADCC和28個ADCE。有一個病例，根據(jù)活檢為BARRETT’S粘膜，而歸為ADCE。ADCC病人的平均年齡為621±136歲，ADCE病人的平均年齡為681±9歲。ADCC中有17個男性，9個女性，而ADCE中有27個男性，1個女性。短期隨訪發(fā)現(xiàn)ADCC病人中有5個有其他腫瘤。在ADCE中只有一個病人有既往腫瘤史。TP53突變31的ADCC和50的ADCE檢測出有TP53突變。兩例ADCE中檢測出兩個突變。突變譜顯示，625ADCC和31?CE的突變是在CPG的C到T轉(zhuǎn)換。用CM1抗體的免疫組化技術(shù)在兩種腫瘤的92中發(fā)現(xiàn)有TP53突變和P53過表達(dá)。MDM2表達(dá)和基因擴(kuò)增50的ADCC有MDM2蛋白表達(dá)（用免疫組化技術(shù)檢測至少10腫瘤細(xì)胞）。用DIFFERENTIALPCR檢測出19的ADCC腫瘤包括MDM2基因擴(kuò)增。40的ADCE有MDM2蛋白表達(dá)（用免疫組化技術(shù)檢測）。細(xì)胞角蛋白4，7，13的表達(dá)癌前病變和癌變常保留了這些細(xì)胞和組織起源的CK表達(dá)類型。在大多數(shù)ADCE（835）和所有ADCC都有CK4表達(dá)。但是，兩種腫瘤在CK7和CK13表達(dá)類型上有差異。50的ADCC為陰性，而80的ADCE為陽性。這些發(fā)現(xiàn)支持ADCC和ADCE是起源于不同的細(xì)胞的假說。

簡介：中文中文3600字出處出處COOPERMA,SONAI,KOMLOSD,ETALLOSSOFEPHRINA5FUNCTIONDISRUPTSLENSFIBERCELLPACKINGANDLEADSTOCATARACTJPROCEEDINGSOFTHENATIONALACADEMYOFSCIENCES,2008,10543166205EPHRINA5功能缺失阻斷晶狀體纖維細(xì)胞包裝導(dǎo)致白內(nèi)障的發(fā)生功能缺失阻斷晶狀體纖維細(xì)胞包裝導(dǎo)致白內(nèi)障的發(fā)生概要概要對于晶狀體纖維細(xì)胞而言，細(xì)胞與細(xì)胞之間的相互作用使他們組成了高度有序的結(jié)構(gòu)來維持透明性。然而這種信號調(diào)節(jié)還沒有被很好的識別。他們在這里報(bào)道的EPHRINA5在晶狀體纖維細(xì)胞的形狀和細(xì)胞間的相互作用中起到了很重要的作用，它是EPH酪氨酸激酶受體的一個配體。與野生型晶狀體纖維細(xì)胞正常的六邊形結(jié)構(gòu)相比，缺乏EPHRINA5功能的小鼠的晶狀體纖維細(xì)胞的橫切面上可以見到細(xì)胞呈圓形或者不規(guī)則。在EPHRINA5缺失的小鼠中，87最后都發(fā)展成了白內(nèi)障，他們又進(jìn)一步證明了EPHRINA5與EPHA2受體之間通過提高ΒCATENIN和N鈣粘蛋白的復(fù)原來調(diào)節(jié)粘附位點(diǎn)復(fù)合結(jié)構(gòu)。引言白內(nèi)障，或者說是晶體混濁化，是在全球范圍內(nèi)造成視覺損傷和失明的第一因素。對于晶狀體發(fā)育和在人的一生中可以維持透明性的分子學(xué)基礎(chǔ)一直不是很明朗。除此之外，對于發(fā)生病理改變的分子和生化的理論也一直不清楚。晶狀體是由位于前表面的一層上皮細(xì)胞構(gòu)成的，在人的一生中，不斷地分裂和分化成下層晶狀體纖維細(xì)胞，因此組成了晶狀體的大部分。在早期的晶狀體發(fā)育中，初級晶狀體纖維細(xì)胞在兩極不斷分化和延長，在隨后的胚胎發(fā)育和整個生命中，次級晶狀體纖維細(xì)胞由赤道板附近的晶狀體上皮細(xì)胞分化而來。在橫切片上可以看到，晶狀體纖維細(xì)胞跟一種向被壓平了的六邊形一樣，兩邊長，剩下四邊很短。這些細(xì)胞被高度有序和密集的組織在一起。而且通過廣泛的細(xì)胞間粘附物，例如縫隙連接，黏附連接。纖維細(xì)胞間隙連接是由連接蛋白（CX）46和50組成，在小鼠中失活后會造成內(nèi)部纖維細(xì)胞變性和白內(nèi)障的發(fā)展。人類連接蛋白基因CX突變也與白內(nèi)障有關(guān)。雖然晶狀體是完全無細(xì)胞，缺血性膜狀物封閉的，但是這些細(xì)胞與細(xì)胞的交界處是提供養(yǎng)分運(yùn)輸，清除代謝廢物的關(guān)鍵，維持動態(tài)平衡。除了縫隙連接，在晶狀體纖維細(xì)胞之間普遍存在著含有N鈣粘蛋白和ΒCATENIN的黏附連接，這些連接可能在晶狀體發(fā)育和功能上起重要作用。雜合型小鼠中卻沒有發(fā)現(xiàn)這樣的變化。從形態(tài)學(xué)和外形尺寸來看，雜合型晶狀體和野生型對照組沒有什么不同。在突變體的晶體中，組織學(xué)分析顯示突變體的晶狀體后囊的破裂和不同嚴(yán)重程度晶狀體破裂。在大多數(shù)非常嚴(yán)重的案例在大多數(shù)非常嚴(yán)重的案例中，殘留下來的組織碎片會對視網(wǎng)膜進(jìn)行撞擊，有時候是對虹膜。我們可以在中，殘留下來的組織碎片會對視網(wǎng)膜進(jìn)行撞擊，有時候是對虹膜。我們可以在這幅圖中看到，野生型的結(jié)構(gòu)很完整，而在突變體重有不同程度組織的破壞所這幅圖中看到，野生型的結(jié)構(gòu)很完整，而在突變體重有不同程度組織的破壞所形成的空泡。形成的空泡。EPHRINA5缺失失去了對細(xì)胞形狀的控制缺失失去了對細(xì)胞形狀的控制為了檢查的缺陷性質(zhì)和初始缺陷的時間，從野生型和PHRINA5缺失小鼠中收集了大量不同發(fā)育階段的晶狀體的切片，并用HE染色。這里選取的是胚胎期第14天,17天,出生當(dāng)天,出生之后第6天,第21天,第30天,和第60天。第一個比較明顯的突變體組織學(xué)改變發(fā)生在P6，一個小空泡出現(xiàn)在皮質(zhì)區(qū)域，但是發(fā)生的比例比較小。引人注目的是，在突變小鼠晶狀體纖維細(xì)胞沒有顯示經(jīng)引人注目的是，在突變小鼠晶狀體纖維細(xì)胞沒有顯示經(jīng)向柱狀組織，也沒有表現(xiàn)出通常出現(xiàn)在野生型晶狀體纖維細(xì)胞中的那種六角密向柱狀組織，也沒有表現(xiàn)出通常出現(xiàn)在野生型晶狀體纖維細(xì)胞中的那種六角密集形態(tài)。而是圓形的空泡狀的細(xì)胞，其中集形態(tài)。而是圓形的空泡狀的細(xì)胞，其中A是由是由HE染色，染色，B是出生后是出生后21天的天的鬼筆環(huán)肽染色。纖維細(xì)胞的長寬比由野生型的鬼筆環(huán)肽染色。纖維細(xì)胞的長寬比由野生型的211降到了突變體中更似立方降到了突變體中更似立方形的形的131嚴(yán)重的晶狀體退化發(fā)生在出生后大約2個月左右的時間。突變型晶狀體纖維細(xì)胞形狀和結(jié)構(gòu)的改變在發(fā)展到任何嚴(yán)重缺陷的形態(tài)之前就可以被

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,THENAATCOULDPROVIDEMORERAPIDDIAGNOSISOFTBANDSUBSEQUENTINITIATIONOFTHERAPYTHISWOULDELIMINATETHENEEDFORINVASIVEDIAGNOSTICPROCEDURES,WHICHARECOSTLYANDPOSEANADDEDRISKTOTHEPATIENTANDALLOWFOREARLIERDISCHARGEOFHOSPITALIZEDPATIENTS25THEREFORE,INADEVELOPINGCOUNTRYSUCHASEGYPT,WECANRESTORETHEUSEOFNAATINSUSPECTCASESWITHAFBSAMPLESTODECREASETHECOSTOFDIAGNOSISTHECOSTOFCULTUREVERSUSSMEAREXAMINATIONSISAROUND25,WHEREASTHECOSTOFTHEGENPROBEPERCASEISAROUND50THEDATAPRESENTEDHERESHOWTHEOUTSTANDINGSENSITIVITYANDSPECIFICITYOFTHEGENPROBETESTSENSITIVITYOFTHEGENPROBETESTWAS967,SPECIFICITYWAS100,POSITIVEPREDICTIVEVALUEWAS100,NEGATIVEPREDICTIVEVALUEWAS952,ANDOVERALLACCURACYWAS98SENSITIVITYWAS80FORSMEARNEGATIVESAMPLESWANGANDTAY25SIMILARLYREPORTEDTHATSENSITIVITYOFAMTDWAS984,SPECIFICITYWAS100,ANDPOSITIVEPREDICTIVEVALUEWAS100INTHEPRESENTSTUDY,THEGENPROBETESTPROVEDTOBEVALUABLEASARAPIDANDACCURATEMETHODFORDIAGNOSISOFPULMONARYTBINCHILDRENATRISKFORTBTHERESULTSCANBEAVAILABLEWITHIN4HOURS,WHEREASTHECULTURERESULTSMAYBEOBTAINEDWITHIN14DAYSWECANNOTDEPENDONZNALONEASARAPIDMETHODBECAUSEOFITSREDUCEDSENSITIVITYTHEREARENUMEROUSDIFFICULTIESINCONTROLLINGTBCONTACTSINTHEEGYPTIANSCENARIOALTHOUGHOFFICIALPROCEDURESRECOMMENDTHATALLCONTACTSOFTBPATIENTSBESCREENED,THELACKOFDIAGNOSTICSTRATEGIESPREVENTSTHEEARLYDIAGNOSISOFALARGENUMBEROFTBCASES,THEREBYINCREASINGDISEASETRANSMISSIONWECANSUGGESTTHEUSEOFCOMBINATIONSOFSIMPLETESTS,SUCHASCHESTXRAYANDZNSTAIN,FORCHILDRENATRISKFORTBFORTHOSEWITHNEGATIVERESULTS,WECANPROCEEDTOCULTUREORNAAT,ACCORDINGTOTHESITUATIONTHERESULTSOFTHISSTUDYSUGGESTTHATTHEGENPROBETESTISANACCURATEMETHODFORRAPIDDETECTIONOFMTUBERCULOSISCOMPLEXESINRESPIRATORYSAMPLESFROMCHILDRENATRISKFORTBITCANBEUSEDFORSMEARNEGATIVECASESTHATARESUSPECTFORTBREFERENCES1ALVESR,SANT’ANNACC,CUNHAAJLAEPIDEMIOLOGIADATUBERCULOSEINFANTILNACIDADEDORIODEJANEIRO,RJREVSAU′DEPU′BLICA200034409–4102AMERICANTHORACICSOCIETYCONTROLOFTUBERCULOSISINTHEUNITEDSTATESAMREVRESPDIS19921461623–16333ABEC,HOSOJIMAS,FUKASAWAY,ETALCOMPARISONOFMBCHECK,BACTEC,ANDEGGBASEDMEDIAFORRECOVERYOFMYCOBACTERIAJCLINMICROBIOL199230878–8814BEAVISKG,LIEHTYMB,VUNGKINDDL,ETALEVALUATIONOFAMPLICORPCRFORDIRECTDETECTIONOFMYCOBACTERIUMTUBERCULOSISFROMSPUTUMSPECIMENSJCLINMICROBIOL1995332582–25865NOORDHOEKGT,MULDERS,WALLACEP,ETALMULTICENTREQUALITYCONTROLSTUDYFORDETECTIONOFMYCOBACTERIUMTUBERCULOSISINCLINICALSAMPLESBYNUCLEICAMPLIFICATIONMETHODSCLINMICROBIOLINFECT200410295–3016RICHELDIL,BARNINIS,SALTINICMOLECULARDIAGNOSISOFTUBERCULOSISEURRESPIRJSUPPL199520689–7007ICHIYAMAS,IINUMAY,YAMORIS,HASEGAWAY,SHIMOKATAK,NAKASHIMANMYCOBACTERIUMGROWTHINDICATORTUBETESTINGINCONJUNCTIONWITHTHEACCUPROBEORTHEAMPLICORPCRASSAYFORDETECTINGANDIDENTIFYINGMYCOBACTERIAFROMSPUTUMSAMPLESJCLINMICROBIOL1997352022–20258ICHIYAMAS,IINUMAY,TAWADAS,ETALEVALUATIONOFGENPROBEAMPLIFIED

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。從那時起MIRNA也廣泛發(fā)現(xiàn)于植物和哺乳動物中。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區(qū)域，而且據(jù)報(bào)道對轉(zhuǎn)錄有正調(diào)控作用。每一個MIRNA都有調(diào)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(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。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在一個樣本中。最近，實(shí)時逆轉(zhuǎn)錄聚合酶鏈反應(yīng)已經(jīng)被應(yīng)用于實(shí)時定量和定性分析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病（ADPKD）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(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)成為一個熱點(diǎn)研究領(lǐng)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(dá)的MIRNA已經(jīng)被證實(shí)，其中29個都下調(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IGA腎病患者活檢標(biāo)本與3個控制組的MIRNA表達(dá)。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(diào)，35個上調(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個下調(diào)，9個上調(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