实例展示如何分析和作图

1、医学生做基础科研，发表医学生做基础科研，发表1-3 分文章的四个套路分文章的四个套路小张聊科研团队小张聊科研团队课程内容一：临床样本出发的研究；一：临床样本出发的研究；二：细胞功能为主的研究；二：细胞功能为主的研究；三：动物模型出发的研究；三：动物模型出发的研究；四：高通量组学为主的研究；四：高通量组学为主的研究； 第三部分第三部分 动物模型出发的研究动物模型出发的研究课程内容一一套路介绍套路介绍二二文章案例解读文章案例解读三三常用的数据库操作演示常用的数据库操作演示四四实例展示如何分析和作图实例展示如何分析和作图五五从结果到成文从结果到成文六六杂志选择杂志选择七七关键问题和注意事项关键问题和

2、注意事项1. Venn图的绘制图的绘制1）Funrich (参见套路一、数据库操作演示（下）参见套路一、数据库操作演示（下）)2. BEG： 3）Venny 2.1： 2. 技术路线图绘制技术路线图绘制Microsoft Visio或者亿图或者亿图Microsoft Visio亿图亿图鱼骨图鱼骨图基本流程图基本流程图3. 机制图机制图Science SlidesNeutrophil Eosinophil BasophilMesenchymal cellSmooth muscle cellsEpithelial cellsMacrophageDendritic cellT-cellErythro

3、cyteB-cellRNA ToolkitMet-tRNAACACDDR1DDR1DDR176123454356217CD45PTP helixVimentinActin and myosinCoiled-coilMicrotubule6 nm10 nm25 nmAntibody Toolkit (IgG)Covalent modification of proteinsUbiquitinationATPUbiquitinUbiquitin activatingenzyme E1Ubiquitin activatingenzyme E1Ubiquitin-conjugatingenzyme E

4、2AMPUbiquitin Ligase E3UbiquitinationsubstrateUbiquitin activatingenzyme E1Ubiquitin-conjugatingenzyme E2UbiquitinationsubstrateWestern Blotting1. Resolve protein samples on SDS-PAGE6. Incubate the blot with chemiluminescent HRP substrate and expose to film 2. Electrophoretically transfer fractionat

5、ed proteins from gel onto PVDF membrane3. Block the membrane with neutral protein (BSA or milk casein)4. Incubate the membrane with primary antibody specific to target protein5. Incubate the membrane with HRP-labeled secondary antibody specific to primary antibodyChromatin Immunoprecipitation Assay

6、(ChIP)1. Crosslink Protein-DNA complexes in situ2. Isolate nuclei and fragment DNA (sonication or digestion)3. Immunoprecipitate with antibody against target nuclear protein and reverse crosslinks4a. Identify protein components of isolated complexes4b. Identify DNA sequence by PCR, cloning and seque

7、ncingEmbryonic stem cellsEgg donorSperm donorEgg fertilizationCulture fertilized cells forseveral cell divisionsDissociate cell clusterfor subculturing andfurther utilizationFreeze in N2 forlong-term storageUse for In VitrofertilizationPotential use fortherapeutic cloningROS in apoptosisCasp 3Caspas

8、e 3 SubstratesApaf1CARDApaf1CARDApaf1CARDCasp 9Casp 9Casp 9CCCCaCaCaCaCaCaCaCaCaIP3RCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaBcl-2BaxBaxCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaCaRYRO-, OH-,NO-BaxBakBaxBakPTPEndoGAIFDiabloCCCCCCCCCCCCCCCCCCEndoGDiabloAIFAIFDiabloCCCApaf1CARDCasp 9SMaseCathepsinsCeramideLysosomeBi

9、dBaxCathepsinp53AIFEndoGGutStomachAdiposetissueHypothalamusPituitaryglandLeptinGhrelinPYYThyroidParathyroidPeptide hormone network in obesityAppetite/weight gain/metabolismsuppression or inductionAdipo-nectinDecreased in obesityIncreased in obesityInsulinPancreasLiver,muscleNPY, AGRPResistinTRH MSH,

10、 GLP-1,CCKGlucagonNormalbronchialepitheliumGoblet cellCiliated cell1 2 3 4 5 6 7 8 9 10 11 12 1314 15 16 17 18 19 20 21 22 X YHuman chromosomesBacteria & Bacteriophage ToolkitBacilliCoccispirillaBacteriophage Bam35Bacteriophage T4Bacteriophage T2The Life cycle of Plasmodium spp.Infected mosquito bitereleases sporozoitesinto human bloodBloodvesselEndotheliumHepatocytesKupffer cellsMerozoiteSpo