泸州科技计划项目申报书-泸州科技计划项目管理平台.doc

”(-) microRNA“”()20141 - 2016122013128(2012)(-) microRNA“”25”646000158083075771958”()”“”35“1miRNA”2345683122”“()“”4532”4936”29”25”24”“”miRNAmiRNA”miRNA”miRNA“”(”) 2 ” ” 100.00 ” 50.00 ”4500”“”“innate immunity”“”“PRR”PAMPsToll“Toll-like receptor, TLR”“PAMPs”CpG DNA“RNA“TLRsPAMPs“88MyD88”TLR-MyD88-IRAK“NF-kB”MAPK”NF-kB-1(AP-1)MyD88”TRIF(TIR)3(IRF3)NF-kBIFN-b”TLR4“”“ TLR26“”TLR2“PRR“”“”miRNAmiRNA”RNARNAmicroRNAmiRNA2l25 ntRNAGRCm381908miRNAGRCh37.p52578miRNA (Sanger miRbase Release 20: June 2013, miRNA”miRNA”miRNAmiRNA“mRNAmiRNADNA”miRNAmRNA”mRNAmiRNA”miRNA”miR-17-5p, miR-20a, miR-106a1, miR155, miR-2231. miRNA” miRNATLR”TLR”Taganov”“”lipopolysaccharide, LPSmiR-146a, miR-132miR-155miR-146aTLRTLRmiR-146a“”“2miR-146miRNA” miR-155LPSTLR-4TLR-212miR146miR-155poly(I:C) (TLR-3)CpGTLR-9”2miR-155LPSPU.1 mRNA “miR-155MyD88TLR”Chen”TLR4 mRNAlet-7TLR4 miRNA”“”“Moschos”LPS“PCR104miRNA“miRNA4.3TNF-”KC2(MIP2)miRNA” 2. miRNA”miR-223”miR-223”“miR-223”Chen“miRNA-223STAT3TLRIL-6IL-1”IL-1miRNA-146a“”miRNA-146aIL-8RANTES” miRNA-146a”miRNA-146a”TLR9miR-146aNotch1IL-12p70TCTL”IRAK2miR-146miR-4661TLRmRNAIL-10 Ma“miR-29IFN-”“Lawless N”(next generation sequencing, NGS)“(Streptococcus uberis, )miRNA21miRNA“miRNA“LPS()145miRNA9”“miRNA”miRNAmiRNA1281 2042miRNA(“)”“”miRNAmiRNAmiRNA”miRNA”miRNASCI“”6miRNA”miRNAmiRNA”miRNA“ ”3500”“”1”1). miRNA“miRNA2). ”miRNA”miRNA”3). miRNA”miRNA2 ”“miRNA1). “D39“24482). “D39“3). “2448 (PBMNALTRNARNARNA“miRNA4). miRNA5). ”RT-PCR“6). ”“2448miRNA”miRNA”/miRNApicTarTargetScanmiRBase Targets“miRNA”miRNA”1. miRNA” (miRNA)1). qPCRmRNA2). Western BlotELISA3). 3UTR/miRNA2. miRNA” (miRNAA )1). “miRNAAA”“2). AmiRNA“miRNAA”“”miRNA”miRNA/”“miRNA“THP-1RAW264.7TC-1”miR mimics, “miR “miR inhibitor, miR sponge“”miR agomirmiR antagomir”TLR”“D39“”miRNA”miRNA1). “D39“4816243240482). “PBM NALTRNA3). ”RT-PCR”miRNA”3.”3.1 ”3.2 ”3.2.1“miRNA1). ”BALB/c”“D3924“4832). “D39“”C+Y(pH7.0)D39”OD620=0.6”PBS5107 CFU/mlBALB/c”41015”“”100L15203). miRNA“2448(NALT)MACSCD14 MicroBeadsmiRNARNAmiRNA”mirVanaTM microRNA Isolation Kit“”“RNaseRNA”-RNA“”25%RNARNA”“55%RNA200ntRNARNARIN)”RNA 6000 Nano KitBioanalyzer 2100 Agilent Small RNA Kit”miRNA.RNA”RNARNAPCRNorthern blot“miRNAmRNA4). miRNA”AffymetrixGeneChip miRNA 3.0“miRNA”Sanger miRbase Release 19miRNAV“2miRNAmiRNA“5). ”real-time RT-PCRNorthern blot, “PCR“”QIAGEN miScript II RT Kit3.2.1 3)miRNAcDNA”miRNA QIAGEN miScript Primer AssaymiScript SYBR Green PCR KitPCR”PCRmiRNA3.2.2”miRNA”1). miRNA”Image Quant 5.0“2miRNA2). miRNA”picTar/miRNAmiRNA“”“”deep sequencing3.2.3 miRNA”1. miRNA”(miRNA)1). mRNA-PCR2). Western BlotELISA3). 3U