實例展示如何分析和作圖

1、醫(yī)學(xué)生做基礎(chǔ)科研，發(fā)表醫(yī)學(xué)生做基礎(chǔ)科研，發(fā)表1-3 分文章的四個套路分文章的四個套路小張聊科研團隊小張聊科研團隊課程內(nèi)容一：臨床樣本出發(fā)的研究；一：臨床樣本出發(fā)的研究；二：細胞功能為主的研究；二：細胞功能為主的研究；三：動物模型出發(fā)的研究；三：動物模型出發(fā)的研究；四：高通量組學(xué)為主的研究；四：高通量組學(xué)為主的研究； 第三部分第三部分 動物模型出發(fā)的研究動物模型出發(fā)的研究課程內(nèi)容一一套路介紹套路介紹二二文章案例解讀文章案例解讀三三常用的數(shù)據(jù)庫操作演示常用的數(shù)據(jù)庫操作演示四四實例展示如何分析和作圖實例展示如何分析和作圖五五從結(jié)果到成文從結(jié)果到成文六六雜志選擇雜志選擇七七關(guān)鍵問題和注意事項關(guān)鍵問題和

2、注意事項1. Venn圖的繪制圖的繪制1）Funrich (參見套路一、數(shù)據(jù)庫操作演示（下）參見套路一、數(shù)據(jù)庫操作演示（下）)2. BEG： 3）Venny 2.1： 2. 技術(shù)路線圖繪制技術(shù)路線圖繪制Microsoft Visio或者億圖或者億圖Microsoft Visio億圖億圖魚骨圖魚骨圖基本流程圖基本流程圖3. 機制圖機制圖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