文献鉴赏ppt课件.ppt

文献鉴赏（一）,1,文献：公开发表的文章或摘要,（1）期刊文章（文献类型标识：J）（2）专著（文献类型标识：M）（3）论文集（文献类型标识：C）,文献类型：,2,基本科学指标数据库（EssentialScienceIndicators，简称ESI）,由世界著名的学术信息出版机构美国科技信息所(ISI)于2001年推出的衡量科学研究绩效、跟踪科学发展趋势的基本分析评价工具，是基于汤森路透WebofScience（SCIE/SSCI）所收录的全球11000多种学术期刊的1000多万条文献记录而建立的计量分析数据库。ESI已成为当今世界范围内普遍用以评价高校、学术机构、国家/地区国际学术水平及影响力的重要评价指标工具之一。,3,美国科学引文索引（ScienceCitationIndex,简称SCI）,于1957年由美国科学信息研究所(ISI)在美国费城创办，1961年创办出版的引文数据库。SCI(科学引文索引)、EI(工程索引)、ISTP(科技会议录索引)是世界著名的三大科技文献检索系统，是国际公认的进行科学统计与科学评价的主要检索工具，其中以SCI最为重要。,影响因子（ImpactFactor,IF),4,中国科学引文数据库（ChineseScienceCitationDatabase，简称CSCD）,创建于1989年，收录我国数学、物理、化学、天文学、地学、生物学、农林科学、医药卫生、工程技术和环境科学等领域出版的中英文科技核心期刊和优秀期刊千余种。中国科学引文数据库具有建库历史最为悠久、专业性强、数据准确规范、检索方式多样、完整、方便等特点，自提供使用以来，深受用户好评，被誉为“中国的SCI”。,5,文献检索常用网站,Pubmed/NCBIWebofScience,文献管理软件：EndNote,6,webofscience,7,8,文献内容框架组成,9,论文题目：Isolationandfunctionalcharacterizationofhumanerythroblastsatdistinctstages:implicationsforunderstandingofnormalanddisorderederythropoiesisinvivo（IF:10.452）,作者：JingpingHu,JingLiu,FuminXue,GregoryHalverson,MarionReid,AnqiGuo,LixiangChen,AzraRaza,NaomiGalili,JulieJaffray,JosephLane,JoelAnneChasis,NaomiTaylor,NarlaMohandasandXiuliAn作者单位：1LaboratoryofMembraneBiology,NewYorkBloodCenter,NewYork,NY;2InstituteofReproductiveandStemCellEngineering,CentralSouthUniversity,Changsha,Hunan,China;3RedCellPhysiologyLaboratory,NewYorkBloodCenter,NewYork,NY;4MolecularBiologyResearchCenter,SchoolofBiologicalScienceandTechnologyandStateKeyLaboratoryofMedicalGeneticsofChina,CentralSouthUniversity,Changsha,Hunan,China;,Blood,2013,121:3246-3253.,10,Introduction,11,ProE:proerythroblasts（原始红细胞）BasoE:basophilicerythroblasts（早幼红细胞）PolyE:polychromaticerythroblasts（中幼红细胞）OrthoE:orthochromaticerythroblasts（晚幼红细胞）,Reticulocyte（网织红细胞）Erythrocyte（红细胞）,12,骨髓增生异常综合征(myelodysplasticsyndromes,MDS)是起源于造血干细胞的一组异质性髓系克隆性疾病，特点是髓系细胞分化及发育异常，表现为无效造血、难治性血细胞减少、造血功能衰竭，高风险向急性髓系白血病（AML）转化。MDS治疗主要解决两大问题：骨髓衰竭及并发症、AML转化。就患者群体而言，MDS患者自然病程和预后的差异性很大，治疗宜个体化。,13,Approachestostudyerythropoiesis,1)Toestablishmethodstoobtainpureerythroidcellsateachdevelopmentalstagefordetailedcellularandmolecularstudies.2)Toestablishmethodstostudyinnormalanddiseasederythropoiesisinvivo.,Normalbonemarrowsmear,14,Mohandas,N,etal.Blood,2008.SalomaoM,etal.PNAS,2008.,Presentviewofstructuralorganizationofnormalredcellmembrane,15,MaterialsandMethods,16,材料与方法,17,Resultsanddiscussion,18,Purificationofhematopoieticstemcells(CD34+cells),19,spectrin,ankyrin,adducin,4.1R,4.9,p55,Tmod,actin,tubulin,GAPDH,Band3,Days68101214161820,Days68101214161820,GLUT1,GPC,GPA,XK,Lu,CD71,a4-integrin,1-integrin,CD44,CD36,Fig1.Expressionofmembraneproteinsduringhumanerythropoiesis,cytoskeletalproteins,surfaceproteins,20,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,101103105,80400,80400,80400,80400,80400,80400,80400,80400,Day6,Day8,Day10,Day12,Day14,Day16,Day18,Day20,GPA,GPC,Band3,RhAG,RhD,b1-integrin,4-integrin,5-integrin,CD44,CD47,CD71,CD147,Kell,101103105,CD59,101103105,CD36,Fig2.Expressionofsurfaceproteinsduringhumanerythropoiesis,21,Fig3.Analysisofdifferentstagesofculturedhumanerythroblastsusinga4-integrinandband3asmarkers,22,Fig4.SeparationofDistinctStagesofErythroblastsusinga4-integrinandBand3asSurfaceMarkers,23,Fig5.SeparationofDistinctStagesofErythroblastsfromPrimaryBoneMarrowusinga4-integrinandBand3asSurfaceMarkers,24,Fig6.AlterederythropoiesisprofileinMDSpatients,25,IsthereanycorrelationbetweenMDSsubtypesandchangesinerythropoiesisprofile,26,文章亮点,Purificationofstagedhumanerythroblastsshouldfacilitateacomprehensivecellularandmolecularcharacterizationofthesecellpopulations.Quantificationofhumanterminalerythropoiesisinvivoprovidesapowerfulmeansforstudyingimpairederythropoiesisinhumandiseases.,27,文章有待改进之处,流式细胞仪数学模型的数字化。病种数和样本数需要增加。,28,CNS系列文章NatureCellScience,29,IsabelleBarde(Firstauthor)DidierTrono(Correspondingauthor)Science,IF31.853,AKRAB/KAP1-miRNACascadeRegulatesErythropoiesisThroughStage-SpecificControlofMitophagy,30,KAP-1,KAP-1是一种转录中介因子,在诸多转录调控复合体中起桥梁作用。大量研究表明,KAP-1作为一个桥梁分子,主要以共抑制因子形式参与转录抑制复合体的形成,在某些复合体中也可作为共激活因子发挥作用。KRPB/KAP1系统在“沉默”哺乳动物基因组反转录因子元件中发挥作用。,31,实验设计,1、材料丰富,32,2、多层次,33,材料与方法,Mouseexperiments.Cells.,Transmissionelectronmicroscopy.RT-QPCR.microRNAexpressionprofilingassay.Chromatinimmunoprecipitation(ChIP)sequencing.RNA-Seq.DirectRNAquantificationbyNanoStringnCounter.,材料,方法,34,Generalstrategy.TotalbonemarrowfromKap-flox/stopfloxYFPCD45.2malemice,withorwithoutaMx-Cretransgene,wasinjectedintolethallyirradiatedCD45.1femalemice,beforepICinjection6-8weekslatertoinducehemato-specificKap1knockout.,35,ComparativehemogramsofwildtypeandKap1KOmice,3weekspost-pICinjection(n=10,*p0.05,*p0.01,*p0.001).Chimerismvariedbetween70and85%.,36,结果,37,Fig1.BlockederythrocytematurationandaccumulationofmitochondriainKap1-deletederythroblasts.,38,Fig2.AKAP1-miRNAcascadecontrolsredcellmitophagy.,39,Fig3.Erythroblast-specificKRAB-ZFPscontrolthemiR-351/Bnip3L/mitophagyaxis.,40,Fig4.KAP1-regulatedRNAinterferencecontrolshumanredcellmitop