新疆褐牛Toll样受体4外显子Ⅲ多态性与奶牛体细胞评分的相关性研究
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摘要
Toll-样受体(Toll like receptors,TLRs)可以通过识别病原微生物启动天然免疫并激活适应性免疫。本研究以中国荷斯坦奶牛和新疆褐牛为研究对象,对TLR4基因外显子3进行了序列分析,并对发现的多态性位点进行PCR-RFLP分析,共发现TLR4 E3+1656、TLR4E3+2021和TLR4 E3+2414三个多态位点,其中TLR4 E3+1656位点发生T/C的碱基突变,但没有引起氨基酸变化;TLR4 E3+2021位点C/T的碱基突变,导致编码氨基酸由苏氨酸变为异亮氨酸;TLR4 E+2414位点发生G/A突变,引起氨基酸由精氨酸变为谷氨酸。但由于TLR4 E+2414位点只发现2个AB基因型个体,其余均为BB基因型个体,不具有统计意义,因此只对TLR4 E3+1656和TLR4E3+2021位点χ2检验表明:两个品种这两个位点的突变均处于Hardy-Weinberg平衡状态(P>0.05)。运用最小二乘法分析表明:TLR4 E3+1656位点的各基因型个体间的平均SCS差异不显著(P>0.05);而TLR4 E3+2021位点中AA与AB基因型SCS水平差异显著(P<0.05),AA与BB基因型SCS水平差异极显著(P<0.01),但AB与BB基因型SCS水平差异不显著(P>0.05),说明TLR4 E3+2021位点的AA基因型可能与抗乳房炎的抗性有关,A等位基因是乳房炎抗性的有利基因。为进一步研究TLR4 E3+2021位点不同基因型对奶牛SCS的影响,采用Quantitative Real-Time PCR技术检测了新疆褐牛不同基因型个体中TLRs信号通路下游的部分基因mRNA的表达水平,结果显示:NF-κB1、NF-κB2和IL-1β基因mRNA表达水平与TLR4 E3+2021位点AB和BB两种基因型差异显著(P<0.05),TNF、TLR4、IL-10、IL-8、MD-2和Myd88基因mRNA表达水平与TLR4 E3+2021位点AB和BB两种基因型差异不显著(P>0.05)。这些结果提示我们TLR4 E3+2021位点碱基的改变可能引起了核内转录因子NF-κB表达量的变化,从而进一步导致IL-1β的mRNA表达量改变。
Toll like receptors (TLRs) recognize pathogen ligands and mediates signaling to initiate innate and adaptive immune responses. In this experiment, Chinese holstei and XinJiang Brown Battle were used as experimental example. The genetic polymorphisms of TLR4 gene exonⅢwere detected by sequencing in the first and by RFLP in the second. The results showed that T/C polymorphic sites did not cause the AA variation in TLR4 E3+1656; C/T polymorphic sites mutation exists in TLR4 E3+2021, which induced that changed Thr into Ile. Statistical results ofχ2 test indicated that two polymorphism sites in the two populations fitted with Hardy–Weinberg equilibrium(P>0.05). The associations were observed between genotypes and SCS by the least square analysis. The results showed that SCS was not significantly affected by different genotypes in TLR4 E3+1656 (P>0.05);both of a significant different effect between genotype AA and AB was found (P<0.05) ,the difference was significant between genotype AA and BB was found (P<0.01) and no significant difference were detected between genotype AB and BB(P>0.05)in TLR4 E3+2021 site. The AA genotype in TLR4 E3+2021 site would be the best genotype to resist mastitis. In short, the allele A might play an important role in mastitis resistance in bovine. To research the effect of different genotypes of TLR4 E3+2021 site on SCS, part of the mRNA of the gene expression where in TLRs signaling pathway downstream were detected by Quantitative Real-Time PCR. The level of NF-κB1、NF-κB2 and IL-1βtranscripts was significantly correlated with genotype of AA and BB in TLR4 E3+2021 (P<0.05),but the level of TNF、TLR4、IL-10、IL-8、MD-2 and Myd88 transcripts was no significantly correlated with genotype of AA and BB in TLR4 E3+2021(P>0.05). These give the implication: The changes of TLR4 E +2021 base cause the change of NF-κB expression, which would lead to the change of IL-1βexpression of mRNA.
Toll like receptors (TLRs) recognize pathogen ligands and mediates signaling to initiate innate and adaptive immune responses. In this experiment, Chinese holstei and XinJiang Brown Battle were used as experimental example. The genetic polymorphisms of TLR4 gene exonⅢwere detected by sequencing in the first and by RFLP in the second. The results showed that T/C polymorphic sites did not cause the AA variation in TLR4 E3+1656; C/T polymorphic sites mutation exists in TLR4 E3+2021, which induced that changed Thr into Ile. Statistical results ofχ2 test indicated that two polymorphism sites in the two populations fitted with Hardy–Weinberg equilibrium(P>0.05). The associations were observed between genotypes and SCS by the least square analysis. The results showed that SCS was not significantly affected by different genotypes in TLR4 E3+1656 (P>0.05);both of a significant different effect between genotype AA and AB was found (P<0.05) ,the difference was significant between genotype AA and BB was found (P<0.01) and no significant difference were detected between genotype AB and BB(P>0.05)in TLR4 E3+2021 site. The AA genotype in TLR4 E3+2021 site would be the best genotype to resist mastitis. In short, the allele A might play an important role in mastitis resistance in bovine. To research the effect of different genotypes of TLR4 E3+2021 site on SCS, part of the mRNA of the gene expression where in TLRs signaling pathway downstream were detected by Quantitative Real-Time PCR. The level of NF-κB1、NF-κB2 and IL-1βtranscripts was significantly correlated with genotype of AA and BB in TLR4 E3+2021 (P<0.05),but the level of TNF、TLR4、IL-10、IL-8、MD-2 and Myd88 transcripts was no significantly correlated with genotype of AA and BB in TLR4 E3+2021(P>0.05). These give the implication: The changes of TLR4 E +2021 base cause the change of NF-κB expression, which would lead to the change of IL-1βexpression of mRNA.
引文
[1]陈北亨.兽医产科学(第二版).北京:农业出版社,1988.
[2] Heringstad B,Klemetsdal G,Ruane J.Clinical Mastitis in Norwegian Cattle:Frequency, Variance Components,and Genetic Correlation with Protein Yield[J]. Livestock Production Science,2000,64:95-106.
[3] Gill,Wayne H Howard,Kemmeth E Leslie,et al. Economics of Mastitis Control[J]. Journal of Dairy Science,73:3340-3348.
[4] Lund T. Genetic relationship between clinical mastitis,somatic cell count,and udder conformation in Danish Holsteins[J].Livestock Production Science,1994,(39):243-251.
[5]谷润林.奶牛乳房炎的病因及其防治[J].内蒙古畜牧科学,2003,2(9):02-03.
[6]李勇,葛秀国,窦忠英.奶牛隐性乳房炎研究进展[J].黄牛杂志,2003,29(3):38-41.
[7]周绪正,张继瑜,王有祥,等“.乳源康”治疗奶牛乳房炎药效学研究[J].中国兽药杂志, 2005,39(8):45-48.
[8]陈国灿,王九峰,马全磊.奶牛隐性乳房炎诊断新方法探讨[J].中国兽医杂志,2005,41 (9): 31-32.
[9]陈家璞.乳牛疾病学[M].北京:农业出版社,1992.
[10]宋继谒.病理学[M].北京:科学出版社.1999.
[11]杨光华.病理学[M].第五版.北京:人民卫生出版社,2001.
[12]林曦.家畜病理学[M].第三版.北京:中国农业出版社,2001.
[13]马学恩.家畜病理学[M].第四版.北京:中国农业出版社,2007.
[14] Lenz A,Franklin GA,Cheadle WG.Systemic inflammation after trauma[J]. Injury,2007,38(12):1336-1345.
[15] Carl Nathan,Michael Sporn.Cytokines in context[J].Journal of Cell Biology,1991,113(5): 981-986.
[16]张荣臻.家畜病理学[M].第三版.内蒙古:内蒙古农牧学院出版,1993.
[17]安庆云.免疫学基础[M].北京:科学出版社,1998.
[18] Lorraine I,Mckay,John A.Cidlowski.CBP(CREB Binding Protein)Integrates NF-kB(Nuclear Factor-kB)and Lucocorticoid Receptor Physical Interactions and Antagonism[J]. Molecular endocrinology,2000,14(8):1222-1234.
[19] Lee JH,Choi JK,Noh MS,et al. Anti-inflammatory effect of kamebakaurin in vivo animal models[J].Planta Medica,2004,70(6):526-530.
[20] Javier Gómez,David García Domingo,Carlos Martínez A,et al. Role of NF-kB in the control of apoptotic and proliferative responses in IL - 2 - responsive T cells [J].Frontiers in Bioscience,1997,15(2):49-60.
[21] McKay L I,Cidlowski J A.MolecuLar control of immune/inflammatory responses:interactions between nuclear factor-kB and steroid receptor-signaling pathways[J]. Endocrine reviews, 1999,20(4):435–459.
[22] GiuLiani C,Napolitano G,Bucci I,et al. NF-kB transcription factor:role in thepathogenesis of inflammatory therapyimp lications[J].Clinica Terapeutica,2001,152(4):249-253.
[23] Jancso G,Cserepes B,Gasz B,et al. Effect of acetylsalicylic acid on nuclearfactor-kappa B activation and on late preconditioning against infarction in the myocardium[J].Journal of CardiovascuLar Pharmacology,2005,46(3):295-301.
[24] Lee JI,Burckart GJ.Nuclear factor kappa B:important transcription factor and therapeutic target[J]. Clin Pharmacol,1998,38(11):981-993.
[25] Watanabe N,Fujita T.NF-κB system[J].Protein Nucleic acid Enzyme,1996,41(l):1198-1209.
[26]郭双平.核转录因子NF-κB的研究进展[J].中华病理学杂志,2000,29(5):379-380.
[27] Blackwell T S,Christman J W.The role of nuclear factor-κB in cytokine gene reguLation[J].American journal of respiratory cell and molecular biology,1997,17(2):3-9.
[28] Jacek Hawiger.Innate Immunity and Inflammation:A Transcriptional Paradigm[J]. Immunologic Research,2001,23(2/3):99–109.
[29] Parry G C N,Mackman N.NF-κB mediated transcription in human monocytic cells and endothelial cells[J]. Trends in cardiovascular medicine,1998,8:138–142.
[30] Lee K M,Kang B S,Lee HL,et al. Spinal NF-κB activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity[J].The European Journal of Neuroscience,2004,19(12):3375-3381.
[31]郭祥君.奶牛乳房炎[J].四川奶牛,2000,2(3):5-6.
[32] Klastrup O,Bakken G,BramLey J,et al. Environmental influences on bovine mastitis[J]. BuLletin of the international dairy federation,1987,217:37.
[33] Watts J L.Etiological agents of bovine mastitis[J].Veterinary microbiology,1988,16(1):41-66.
[34]连学昭.防治奶牛乳房炎新型中药制剂的研究[D].大庆:黑龙江八一农垦大学,2006.
[35]袁永隆,张礼华,刘纯传,等.我国奶牛乳房炎常见病原菌的区系调查[J].中国农业科学,1992,25(4):70-76.
[36]伍义行,黄利权.奶牛乳腺炎防治的免疫学和药理学机制[J].中国兽医杂志,2003,39(3):40-44.
[37]孙庆华,赵宗胜.进口荷斯坦牛隐性乳腺炎病原菌的分离鉴定[J].乳业科学与技术,2006,1: 33-35.
[38]李宏胜,郁杰,罗金印.我国部分地区个体奶牛场乳房炎细菌学调查[J].中兽医医药杂志, 2002,1(6):15-16.
[39] Emanuelson U,Danell B,Philipsson J.Genetic Parameters for Clinical Mastitis,Somatic Cell Counts,and Milk Production Estimated by Multiple-Trait Restricted Maximum Likelihood[J]. Journal of dairy science,1998,71(2):467-476.
[40] Poso J,Mantysaari E A.Relationships between Clinical Mastitis,Somatic Cell Score,and Production for the First Three Lactations of Finnish Ayrshire[J]. Journal of dairy science,1996, 79(7):1284-1291.
[41] Heringstad B,Klemetsdal G,Ruane J.Clinical Mastitis in Norwegian Cattle:Frequency,Variance Components,and Genetic Correlation with Protein Yield[J]. Journal of dairy science,1999,82(6):1325-1330.
[42] Strandberg E,Shook G E.Genetic and Economic Responses to Breeding Programs that Consider Mastitis. Journal of dairy science,1989,72(8):2136-2142.
[43] Rupp R,Boichard D.Genetic Parameters for Clinical Mastitis,Somatic Cell Score,Production, Udder Type Traits,and Milking Ease in First Lactation Holsteins[J]. Journal of dairy science,1999,82(10):2198-2204.
[44] Lund T,Miglior F,Dekkers J C,et al. Genetic Relationships between Clinical Mastitis Somatic Cell Count,and Udder Conformation in Danish Holsteins[J]. Livestock production science,1994,39(4):243-251.
[45] Turner A G,Salmonsen P A.The effect of relative humidity on the survival of three serotypes of Klebsiella[J].Journal of Applied Bacteriology,1973,36(3):497-499.
[46] Birm M,Timms L.In vitro growth of environmental mastitis pathogens in various bedding materials[J].Canadian Journal of Animal Science,1989,72(2):287-297.
[47] Emanuelson U,Danell B,Philipsson J.Genetic Parameters for Clinical Mastitis,Somatic Cell Counts and Milk Production Estimated by MuLtiple-Trait Restricted Maximum Likelihood[J]. Journal of dairy science,1998,71:467-476.
[48] Machado J C, Figueiredo C, Canedo P,et al.A proinflammatory genetic profile increases the risk for chronic atrophic gastritis and gastric carcinoma[J].Gastroenterology,2003,125(2):364-371.
[49] Hibbitt G,Cole B,Reiter.Antimicrobial proteins isolated from the teat canal of the cow[J]. Journal of general microbiology,1969,56(3):365-371.
[50] Lam T J,van Vliet J H,Schukken Y H. Udder disinfection and mastitis in cattle:a literature review [J]. Tijdschrift voor diergeneeskunde,1995,120(13):392-399.
[51]伍义行,黄利权.奶牛乳房炎防治的免疫学和药理学机制[J].中国兽医杂志,2003,39(3): 40-42.
[52] Sutra L,Poutrel B.ViruLence factors involved in the pathogenesis of bovine intramammary infections due to Staphylococcus aureus[J]. Journal of medical microbiology,1994,40(2):79-89.
[53] McGuirk P,Mills K H,Mills.Pathogen-specific regulatory T cells provoke a shift in the Th1/Th2 paradigm in immunity to infectious diseases[J]. Trends Immunol,2002,23(9): 450-455.
[54] Mrode R A,Swanson G J.Genetic and statistical properties of somatic cell count and its suitability as an indireet means of redueing the ineidenee ofmatitis in dairy cattle [J].Animal Breeding Abstraets,1996,64(11):847-857.
[55]郝建国,梁淑萍.日本学者对体细胞数与乳房炎关系论述[J].中国奶牛,2002,3(1):52-54.
[56]郑州市奶牛乳房炎调查组.郑州市奶牛隐性乳房炎的调查报告[J].河南农业科学,1988,8(4):29-31.
[57]萧乾庆,冯端明,王榕,等.奶牛隐性乳房炎的诊治试验[J].中国奶牛,1997,2(4):17-18.
[58] Bes M,Guerin Faublee V,Meugnier H,et al.Improvement of the identification of staphylococci isolated from bovine mammary infections using molecular methods [J]. Veterinary microbiology,2000,71(3):287-294.
[59] Sabot G,Rose P,Hickey W J,e et al.Selective and sensitive method for PCR amplification of Escherichia coli 16S rRNA genes in soil[J]. Applied and environmental microbiology,2000,66(2):844-849.
[60] Straub J A,Hertel C,Hammes W P.A 23S rDNA-targeted polymerase chain reaction based system for detection of Staphylococcus aureus in meat starter cuLtures and dairy products[J]. Journal of food protection,1999,62(10):1l50-l156.
[61]贾玉萍,周东顺,万仁忠,等.巢式PCR检测无乳链球菌16SrRNA方法的建立及应用[J].农业生物技术学报,2005,13(5):668-671.
[62]谢志勤,谢芝勋,唐小飞等.多重PCR快速检测奶牛乳房炎3种主要病原体[J].畜牧与兽医, 2006,38(7):4-7.
[63] Ali A,Shook G E.An optimum transformation for somatic cell concentration in milk[J]. Journal of Dairy Science,1980,63(3):487-490.
[64] Samore A B.Genetic aspects of somatic cell count in the Italian Holstein friesian popuLation [D],Wageningen University.2003.
[65] Shook G E,Schutz M M. Selection on somatic cell score to improve resistance to mastitis in the United States [J]. Journal of Dairy Science,1994,77(2):648-658.
[66] Shook G E. Genetic Improvement of Mastitis Through Selection on Somatic Cell Count, Veterinary Clinics of North America[J].Food Animal Practice,1993,9(3):563-581.
[67] DeGroot B J, Keown J F, Van Vleck L D,et al.Genetic parameters and responses of linear type, yield traits, and somatic cell scores to divergent selection for predicted transmitting ability for type in Holsteins [J]. Journal of dairy science,2002,85(6):1578-1585.
[68] Rothschild M F,Soller M.Candidate gene analysis to deteet genes controlling traits of economic importance in domestic livestoek[J].Probe,1997,8(l):13-20.
[69] Ashwell M S,Rexroad CE,Miller R H,et al.Mapping economie trait loci for somatic cell score in Holstein cattle using microsatellite marker sand seleetive genotyping [J].Animal Genetics, 1996,27(4):235-242.
[70] Medzhitov R,Preston H P,Janeway C A.A human homologue of the Drosophlia Toll protein singals activation of adaptive immunity[J].Nature,1997,388(6640):394-397.
[71] Jin M S,Lee J O. Structures of TLR-ligand complexes[J].Current opinion in immunology,2008,20(4):414-419.
[72] Jin M S,Lee J O. Structures of the toll-like receptor family and its ligand complexes[J]. Immunity,2008,29(2):182-191.
[73] Cheung P C,Nebreda A R,Cohen P.TAB3,a new binding partner of the protein kinase TAK1 [J].The Biochemical journal,2004,378(1):27-34.
[74] Jin G,Klika A,Callahan M,et al.Identification of a human NF-kappaB activating protein, TAB3[J].Proc Natl Acad Sci USA,2004,101(7):2028-2033.
[75] Kanayama A,Seth R B,Sun L,et al.TAB2 and TAB3 activate the NF-kappaB pathway through binding to polyubiquitin chains[J].Molecular cell,2004,15(4):535-548.
[76] Ea C K,Sun L,Inoue J,et al.TIFA activates IkappaB kinas(eIKK)by promoting oligomerization and ubiquitination of TRAF6[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(43):15318-15323.
[77] Muzio M,Bosisio D,Polentarutti N,et al.Differential expression and reguLation of toll-like receptors(TLR)in human leukocytes:selective expression of TLR3 in dendritc cells[J].Joumal of Iinmunology,2000,164(11):5998-6004.
[78] Chuang T H,Ulevitch R J. Triad3A,an E3 ubiquitin-protein ligase regulating Toll-like receptors[J].Nature immunology,2004,5(5):495—502.
[79] McGuire K,Jones M,Werling D,et al.Radiation hybrid mapping of all 10 characterized bovine Toll-like receptors[J].Animal Genetics,2006,37(1):47-50.
[80] Chapes S K,Mosier D A,Wiright A D,et al.TLR4 and Nramp1 genes control host pulmonary resistance against the opportunistic bacterium Pasteurella pneumotropica[J].Journal of leukocyte biology,2001,69(3):381-386.
[81] Puthothu B,Forster J,Heinzmann A,et al.TLR-4 and CD14 polymorphisms in respiratory syncytial virus associated disease[J].Dis Markers,2006,22(5-6):303-308.
[82] Beutler B,Poltorak A.Sepsis and evolution of the innate immune response[J].Critical care medicine,2001,29(7):2-7.
[83]李传友.TLR受体在抗结核免疫中的作用[J].中国防痨杂志,2006,18(2):18.
[84]奉俊敏,刘运海,申向民.急性脑梗死患者TLR4 mRNA表达与TNF-α相关性研究[J].中华神经医学杂志,2006,5(2):149-151.
[85] Wang X,Athayde N,Trudinger B.Placental vascuLar disease and toll-like receptor 4 gene expression[J].American journal of obstetrics and gynecology,2005,192(3):961-966.
[86]许志良,刘永林.糖尿病患者CD14细胞TLR2、HLA2DR的表达[J].浙江中西医结合杂志,2006,16(6):350-351.
[87] Candia L,Marquez J,Hernandez C,et al.Toll-like receptor-2 expression is upregu -lated inantigen-presenting cells from patients with psoriatic arthritis:a pathogenic role for innate immunity[J].Rheumatol,2007,34(2):374-379.
[88] Sarah S M,Tamilselvan S,Kamatchiammal S,et al.Expression of Toll-like receptors 2 and 4 ingingivitis and chronic periodontitis[J].Indian journal of dental research,2006,17(3):114-116.
[89] Beuder B.Toll-like receptors:how they work and what they do[J]. Current opinion in hematology,2002,9(1):2-10.
[90]李妍,宁云山.Toll样受体与抗感染免疫[J].国外医学免疫学分册,2004,27(4):228-230.
[91] B R Achyut . Association of Toll-like receptor-4(Asp299Gly and Thr399Ileu)gene polymorphisms with gastritis and precancerous lesions[J].Human immunology,2007,10(6):1-7.
[92] Poltorak A,Hex,Sminroval,et al.Defective LPS signalingin C3H/HeJ and C57BL/ 10ScCr mice:mutationsin TLR4 gene[J].Science,1998,282(5396):2085-2088.
[93] Sharma b S, Leyyai,Schenkelf, et al.Association of Toll-like receptor 4 polymorphisms with somatic cell scoreand lactation persistency in holstein buLls[J].Journal of Dairy Science,2006,89(9):3626-3635.
[94] Goldannner T,Zerbe H,Molenaar A,et al.Mastitis increase mannnary mRNA abundance ofβ-defensin 5,Toll-like-receptor 2(TLR2),and TLR4 but not TLR9 in cattle [J].Clinical and Diagnostic Laboratory Immunology,2004,11(l):174-185.
[95] Higuchi R,Fockler C,et al.Kinetic PCR analysis:Real-Time monitoring of DNA amplification reactions[J].Biotechnology,1993,11(9):1026-1030.
[96] GiuLictti A,Ovrbergh L,Valckx D,et al.An overview of real-time quantitative PCR: applications to quantify cytokine gene expression [J].Mwthods,2001,25(4):386-401.
[97] Shizuo A,Kiyoshi T,Tsuneyasu K.Toll-like receptors:critical proteins linking innate and acquired immunity [J]. Nature immunology,2001,2(8):675-680.
[98]黄培堂等译.分子克隆试验指南(第三版)[M].北京:科学技术出版社,2002.
[99] Wang Xingping,Xu Shangzhong,Gao Xue,et al.Genetic Polymorphism of TLR4 Gene and Correlation with Mastitis in Cattle[J].Journal of Genetics and Genomics,2007,34(5):406-412.
[100]刘文娇,孙少华,李雪梅. TLR4基因外显子ⅢBbv12 I酶切位点多态性与奶牛乳房炎的相关分析[J].中国农业科技导报,2008,10(2):114-118.
[101] Heringstad B,Gianola D,Chang Y M,et al.Genetic associations between clinical mastitis and somatic cell score in early first-lactation cows[J]. Journal of Dairy Science. 2006,89(6):2236–2244.
[102] Monardes H G,J F Hayes.Genetic and phenotypic relationships between lactation cell counts and milk yield and composition of Holstein cows[J]. Journal of Dairy Science,1985,68(5):1250–1256.
[2] Heringstad B,Klemetsdal G,Ruane J.Clinical Mastitis in Norwegian Cattle:Frequency, Variance Components,and Genetic Correlation with Protein Yield[J]. Livestock Production Science,2000,64:95-106.
[3] Gill,Wayne H Howard,Kemmeth E Leslie,et al. Economics of Mastitis Control[J]. Journal of Dairy Science,73:3340-3348.
[4] Lund T. Genetic relationship between clinical mastitis,somatic cell count,and udder conformation in Danish Holsteins[J].Livestock Production Science,1994,(39):243-251.
[5]谷润林.奶牛乳房炎的病因及其防治[J].内蒙古畜牧科学,2003,2(9):02-03.
[6]李勇,葛秀国,窦忠英.奶牛隐性乳房炎研究进展[J].黄牛杂志,2003,29(3):38-41.
[7]周绪正,张继瑜,王有祥,等“.乳源康”治疗奶牛乳房炎药效学研究[J].中国兽药杂志, 2005,39(8):45-48.
[8]陈国灿,王九峰,马全磊.奶牛隐性乳房炎诊断新方法探讨[J].中国兽医杂志,2005,41 (9): 31-32.
[9]陈家璞.乳牛疾病学[M].北京:农业出版社,1992.
[10]宋继谒.病理学[M].北京:科学出版社.1999.
[11]杨光华.病理学[M].第五版.北京:人民卫生出版社,2001.
[12]林曦.家畜病理学[M].第三版.北京:中国农业出版社,2001.
[13]马学恩.家畜病理学[M].第四版.北京:中国农业出版社,2007.
[14] Lenz A,Franklin GA,Cheadle WG.Systemic inflammation after trauma[J]. Injury,2007,38(12):1336-1345.
[15] Carl Nathan,Michael Sporn.Cytokines in context[J].Journal of Cell Biology,1991,113(5): 981-986.
[16]张荣臻.家畜病理学[M].第三版.内蒙古:内蒙古农牧学院出版,1993.
[17]安庆云.免疫学基础[M].北京:科学出版社,1998.
[18] Lorraine I,Mckay,John A.Cidlowski.CBP(CREB Binding Protein)Integrates NF-kB(Nuclear Factor-kB)and Lucocorticoid Receptor Physical Interactions and Antagonism[J]. Molecular endocrinology,2000,14(8):1222-1234.
[19] Lee JH,Choi JK,Noh MS,et al. Anti-inflammatory effect of kamebakaurin in vivo animal models[J].Planta Medica,2004,70(6):526-530.
[20] Javier Gómez,David García Domingo,Carlos Martínez A,et al. Role of NF-kB in the control of apoptotic and proliferative responses in IL - 2 - responsive T cells [J].Frontiers in Bioscience,1997,15(2):49-60.
[21] McKay L I,Cidlowski J A.MolecuLar control of immune/inflammatory responses:interactions between nuclear factor-kB and steroid receptor-signaling pathways[J]. Endocrine reviews, 1999,20(4):435–459.
[22] GiuLiani C,Napolitano G,Bucci I,et al. NF-kB transcription factor:role in thepathogenesis of inflammatory therapyimp lications[J].Clinica Terapeutica,2001,152(4):249-253.
[23] Jancso G,Cserepes B,Gasz B,et al. Effect of acetylsalicylic acid on nuclearfactor-kappa B activation and on late preconditioning against infarction in the myocardium[J].Journal of CardiovascuLar Pharmacology,2005,46(3):295-301.
[24] Lee JI,Burckart GJ.Nuclear factor kappa B:important transcription factor and therapeutic target[J]. Clin Pharmacol,1998,38(11):981-993.
[25] Watanabe N,Fujita T.NF-κB system[J].Protein Nucleic acid Enzyme,1996,41(l):1198-1209.
[26]郭双平.核转录因子NF-κB的研究进展[J].中华病理学杂志,2000,29(5):379-380.
[27] Blackwell T S,Christman J W.The role of nuclear factor-κB in cytokine gene reguLation[J].American journal of respiratory cell and molecular biology,1997,17(2):3-9.
[28] Jacek Hawiger.Innate Immunity and Inflammation:A Transcriptional Paradigm[J]. Immunologic Research,2001,23(2/3):99–109.
[29] Parry G C N,Mackman N.NF-κB mediated transcription in human monocytic cells and endothelial cells[J]. Trends in cardiovascular medicine,1998,8:138–142.
[30] Lee K M,Kang B S,Lee HL,et al. Spinal NF-κB activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity[J].The European Journal of Neuroscience,2004,19(12):3375-3381.
[31]郭祥君.奶牛乳房炎[J].四川奶牛,2000,2(3):5-6.
[32] Klastrup O,Bakken G,BramLey J,et al. Environmental influences on bovine mastitis[J]. BuLletin of the international dairy federation,1987,217:37.
[33] Watts J L.Etiological agents of bovine mastitis[J].Veterinary microbiology,1988,16(1):41-66.
[34]连学昭.防治奶牛乳房炎新型中药制剂的研究[D].大庆:黑龙江八一农垦大学,2006.
[35]袁永隆,张礼华,刘纯传,等.我国奶牛乳房炎常见病原菌的区系调查[J].中国农业科学,1992,25(4):70-76.
[36]伍义行,黄利权.奶牛乳腺炎防治的免疫学和药理学机制[J].中国兽医杂志,2003,39(3):40-44.
[37]孙庆华,赵宗胜.进口荷斯坦牛隐性乳腺炎病原菌的分离鉴定[J].乳业科学与技术,2006,1: 33-35.
[38]李宏胜,郁杰,罗金印.我国部分地区个体奶牛场乳房炎细菌学调查[J].中兽医医药杂志, 2002,1(6):15-16.
[39] Emanuelson U,Danell B,Philipsson J.Genetic Parameters for Clinical Mastitis,Somatic Cell Counts,and Milk Production Estimated by Multiple-Trait Restricted Maximum Likelihood[J]. Journal of dairy science,1998,71(2):467-476.
[40] Poso J,Mantysaari E A.Relationships between Clinical Mastitis,Somatic Cell Score,and Production for the First Three Lactations of Finnish Ayrshire[J]. Journal of dairy science,1996, 79(7):1284-1291.
[41] Heringstad B,Klemetsdal G,Ruane J.Clinical Mastitis in Norwegian Cattle:Frequency,Variance Components,and Genetic Correlation with Protein Yield[J]. Journal of dairy science,1999,82(6):1325-1330.
[42] Strandberg E,Shook G E.Genetic and Economic Responses to Breeding Programs that Consider Mastitis. Journal of dairy science,1989,72(8):2136-2142.
[43] Rupp R,Boichard D.Genetic Parameters for Clinical Mastitis,Somatic Cell Score,Production, Udder Type Traits,and Milking Ease in First Lactation Holsteins[J]. Journal of dairy science,1999,82(10):2198-2204.
[44] Lund T,Miglior F,Dekkers J C,et al. Genetic Relationships between Clinical Mastitis Somatic Cell Count,and Udder Conformation in Danish Holsteins[J]. Livestock production science,1994,39(4):243-251.
[45] Turner A G,Salmonsen P A.The effect of relative humidity on the survival of three serotypes of Klebsiella[J].Journal of Applied Bacteriology,1973,36(3):497-499.
[46] Birm M,Timms L.In vitro growth of environmental mastitis pathogens in various bedding materials[J].Canadian Journal of Animal Science,1989,72(2):287-297.
[47] Emanuelson U,Danell B,Philipsson J.Genetic Parameters for Clinical Mastitis,Somatic Cell Counts and Milk Production Estimated by MuLtiple-Trait Restricted Maximum Likelihood[J]. Journal of dairy science,1998,71:467-476.
[48] Machado J C, Figueiredo C, Canedo P,et al.A proinflammatory genetic profile increases the risk for chronic atrophic gastritis and gastric carcinoma[J].Gastroenterology,2003,125(2):364-371.
[49] Hibbitt G,Cole B,Reiter.Antimicrobial proteins isolated from the teat canal of the cow[J]. Journal of general microbiology,1969,56(3):365-371.
[50] Lam T J,van Vliet J H,Schukken Y H. Udder disinfection and mastitis in cattle:a literature review [J]. Tijdschrift voor diergeneeskunde,1995,120(13):392-399.
[51]伍义行,黄利权.奶牛乳房炎防治的免疫学和药理学机制[J].中国兽医杂志,2003,39(3): 40-42.
[52] Sutra L,Poutrel B.ViruLence factors involved in the pathogenesis of bovine intramammary infections due to Staphylococcus aureus[J]. Journal of medical microbiology,1994,40(2):79-89.
[53] McGuirk P,Mills K H,Mills.Pathogen-specific regulatory T cells provoke a shift in the Th1/Th2 paradigm in immunity to infectious diseases[J]. Trends Immunol,2002,23(9): 450-455.
[54] Mrode R A,Swanson G J.Genetic and statistical properties of somatic cell count and its suitability as an indireet means of redueing the ineidenee ofmatitis in dairy cattle [J].Animal Breeding Abstraets,1996,64(11):847-857.
[55]郝建国,梁淑萍.日本学者对体细胞数与乳房炎关系论述[J].中国奶牛,2002,3(1):52-54.
[56]郑州市奶牛乳房炎调查组.郑州市奶牛隐性乳房炎的调查报告[J].河南农业科学,1988,8(4):29-31.
[57]萧乾庆,冯端明,王榕,等.奶牛隐性乳房炎的诊治试验[J].中国奶牛,1997,2(4):17-18.
[58] Bes M,Guerin Faublee V,Meugnier H,et al.Improvement of the identification of staphylococci isolated from bovine mammary infections using molecular methods [J]. Veterinary microbiology,2000,71(3):287-294.
[59] Sabot G,Rose P,Hickey W J,e et al.Selective and sensitive method for PCR amplification of Escherichia coli 16S rRNA genes in soil[J]. Applied and environmental microbiology,2000,66(2):844-849.
[60] Straub J A,Hertel C,Hammes W P.A 23S rDNA-targeted polymerase chain reaction based system for detection of Staphylococcus aureus in meat starter cuLtures and dairy products[J]. Journal of food protection,1999,62(10):1l50-l156.
[61]贾玉萍,周东顺,万仁忠,等.巢式PCR检测无乳链球菌16SrRNA方法的建立及应用[J].农业生物技术学报,2005,13(5):668-671.
[62]谢志勤,谢芝勋,唐小飞等.多重PCR快速检测奶牛乳房炎3种主要病原体[J].畜牧与兽医, 2006,38(7):4-7.
[63] Ali A,Shook G E.An optimum transformation for somatic cell concentration in milk[J]. Journal of Dairy Science,1980,63(3):487-490.
[64] Samore A B.Genetic aspects of somatic cell count in the Italian Holstein friesian popuLation [D],Wageningen University.2003.
[65] Shook G E,Schutz M M. Selection on somatic cell score to improve resistance to mastitis in the United States [J]. Journal of Dairy Science,1994,77(2):648-658.
[66] Shook G E. Genetic Improvement of Mastitis Through Selection on Somatic Cell Count, Veterinary Clinics of North America[J].Food Animal Practice,1993,9(3):563-581.
[67] DeGroot B J, Keown J F, Van Vleck L D,et al.Genetic parameters and responses of linear type, yield traits, and somatic cell scores to divergent selection for predicted transmitting ability for type in Holsteins [J]. Journal of dairy science,2002,85(6):1578-1585.
[68] Rothschild M F,Soller M.Candidate gene analysis to deteet genes controlling traits of economic importance in domestic livestoek[J].Probe,1997,8(l):13-20.
[69] Ashwell M S,Rexroad CE,Miller R H,et al.Mapping economie trait loci for somatic cell score in Holstein cattle using microsatellite marker sand seleetive genotyping [J].Animal Genetics, 1996,27(4):235-242.
[70] Medzhitov R,Preston H P,Janeway C A.A human homologue of the Drosophlia Toll protein singals activation of adaptive immunity[J].Nature,1997,388(6640):394-397.
[71] Jin M S,Lee J O. Structures of TLR-ligand complexes[J].Current opinion in immunology,2008,20(4):414-419.
[72] Jin M S,Lee J O. Structures of the toll-like receptor family and its ligand complexes[J]. Immunity,2008,29(2):182-191.
[73] Cheung P C,Nebreda A R,Cohen P.TAB3,a new binding partner of the protein kinase TAK1 [J].The Biochemical journal,2004,378(1):27-34.
[74] Jin G,Klika A,Callahan M,et al.Identification of a human NF-kappaB activating protein, TAB3[J].Proc Natl Acad Sci USA,2004,101(7):2028-2033.
[75] Kanayama A,Seth R B,Sun L,et al.TAB2 and TAB3 activate the NF-kappaB pathway through binding to polyubiquitin chains[J].Molecular cell,2004,15(4):535-548.
[76] Ea C K,Sun L,Inoue J,et al.TIFA activates IkappaB kinas(eIKK)by promoting oligomerization and ubiquitination of TRAF6[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(43):15318-15323.
[77] Muzio M,Bosisio D,Polentarutti N,et al.Differential expression and reguLation of toll-like receptors(TLR)in human leukocytes:selective expression of TLR3 in dendritc cells[J].Joumal of Iinmunology,2000,164(11):5998-6004.
[78] Chuang T H,Ulevitch R J. Triad3A,an E3 ubiquitin-protein ligase regulating Toll-like receptors[J].Nature immunology,2004,5(5):495—502.
[79] McGuire K,Jones M,Werling D,et al.Radiation hybrid mapping of all 10 characterized bovine Toll-like receptors[J].Animal Genetics,2006,37(1):47-50.
[80] Chapes S K,Mosier D A,Wiright A D,et al.TLR4 and Nramp1 genes control host pulmonary resistance against the opportunistic bacterium Pasteurella pneumotropica[J].Journal of leukocyte biology,2001,69(3):381-386.
[81] Puthothu B,Forster J,Heinzmann A,et al.TLR-4 and CD14 polymorphisms in respiratory syncytial virus associated disease[J].Dis Markers,2006,22(5-6):303-308.
[82] Beutler B,Poltorak A.Sepsis and evolution of the innate immune response[J].Critical care medicine,2001,29(7):2-7.
[83]李传友.TLR受体在抗结核免疫中的作用[J].中国防痨杂志,2006,18(2):18.
[84]奉俊敏,刘运海,申向民.急性脑梗死患者TLR4 mRNA表达与TNF-α相关性研究[J].中华神经医学杂志,2006,5(2):149-151.
[85] Wang X,Athayde N,Trudinger B.Placental vascuLar disease and toll-like receptor 4 gene expression[J].American journal of obstetrics and gynecology,2005,192(3):961-966.
[86]许志良,刘永林.糖尿病患者CD14细胞TLR2、HLA2DR的表达[J].浙江中西医结合杂志,2006,16(6):350-351.
[87] Candia L,Marquez J,Hernandez C,et al.Toll-like receptor-2 expression is upregu -lated inantigen-presenting cells from patients with psoriatic arthritis:a pathogenic role for innate immunity[J].Rheumatol,2007,34(2):374-379.
[88] Sarah S M,Tamilselvan S,Kamatchiammal S,et al.Expression of Toll-like receptors 2 and 4 ingingivitis and chronic periodontitis[J].Indian journal of dental research,2006,17(3):114-116.
[89] Beuder B.Toll-like receptors:how they work and what they do[J]. Current opinion in hematology,2002,9(1):2-10.
[90]李妍,宁云山.Toll样受体与抗感染免疫[J].国外医学免疫学分册,2004,27(4):228-230.
[91] B R Achyut . Association of Toll-like receptor-4(Asp299Gly and Thr399Ileu)gene polymorphisms with gastritis and precancerous lesions[J].Human immunology,2007,10(6):1-7.
[92] Poltorak A,Hex,Sminroval,et al.Defective LPS signalingin C3H/HeJ and C57BL/ 10ScCr mice:mutationsin TLR4 gene[J].Science,1998,282(5396):2085-2088.
[93] Sharma b S, Leyyai,Schenkelf, et al.Association of Toll-like receptor 4 polymorphisms with somatic cell scoreand lactation persistency in holstein buLls[J].Journal of Dairy Science,2006,89(9):3626-3635.
[94] Goldannner T,Zerbe H,Molenaar A,et al.Mastitis increase mannnary mRNA abundance ofβ-defensin 5,Toll-like-receptor 2(TLR2),and TLR4 but not TLR9 in cattle [J].Clinical and Diagnostic Laboratory Immunology,2004,11(l):174-185.
[95] Higuchi R,Fockler C,et al.Kinetic PCR analysis:Real-Time monitoring of DNA amplification reactions[J].Biotechnology,1993,11(9):1026-1030.
[96] GiuLictti A,Ovrbergh L,Valckx D,et al.An overview of real-time quantitative PCR: applications to quantify cytokine gene expression [J].Mwthods,2001,25(4):386-401.
[97] Shizuo A,Kiyoshi T,Tsuneyasu K.Toll-like receptors:critical proteins linking innate and acquired immunity [J]. Nature immunology,2001,2(8):675-680.
[98]黄培堂等译.分子克隆试验指南(第三版)[M].北京:科学技术出版社,2002.
[99] Wang Xingping,Xu Shangzhong,Gao Xue,et al.Genetic Polymorphism of TLR4 Gene and Correlation with Mastitis in Cattle[J].Journal of Genetics and Genomics,2007,34(5):406-412.
[100]刘文娇,孙少华,李雪梅. TLR4基因外显子ⅢBbv12 I酶切位点多态性与奶牛乳房炎的相关分析[J].中国农业科技导报,2008,10(2):114-118.
[101] Heringstad B,Gianola D,Chang Y M,et al.Genetic associations between clinical mastitis and somatic cell score in early first-lactation cows[J]. Journal of Dairy Science. 2006,89(6):2236–2244.
[102] Monardes H G,J F Hayes.Genetic and phenotypic relationships between lactation cell counts and milk yield and composition of Holstein cows[J]. Journal of Dairy Science,1985,68(5):1250–1256.