簡(jiǎn)介：浙江大學(xué)碩士學(xué)位論文蛋氨酸、賴氨酸及其二肽對(duì)奶牛乳腺上皮細(xì)胞酪蛋白Α基因表達(dá)的影響姓名楊金勇申請(qǐng)學(xué)位級(jí)別碩士專業(yè)動(dòng)物營(yíng)養(yǎng)與飼料科學(xué)指導(dǎo)教師劉建新吳躍明200605012006年浙江大學(xué)碩士學(xué)位論文致謝本論文是在導(dǎo)師劉建新教授悉心指導(dǎo)下得以順利完成的。值此碩士學(xué)位論文完成之際，謹(jǐn)向三年來傾心指導(dǎo)我的導(dǎo)師劉建新教授致以最誠(chéng)摯的敬意與衷心的感謝。在攻讀碩士學(xué)位期間，深深感受到導(dǎo)師淵博的知識(shí)、博大的胸懷、一絲不茍的工作作風(fēng)、嚴(yán)謹(jǐn)?shù)闹螌W(xué)態(tài)度、忘我的工作精神和創(chuàng)新的科學(xué)理念，這些都時(shí)刻激勵(lì)著我并將成為我受益終身的寶貴財(cái)富。吳躍明教授在試驗(yàn)設(shè)計(jì)和實(shí)施過程中給予了精心指導(dǎo)，葉均安副教授在為人處世方面給了我極大的影響，張才喬教授在細(xì)胞培養(yǎng)實(shí)驗(yàn)中給了我細(xì)心的指導(dǎo)和幫助，在這里學(xué)生向各位老師表示深深的感謝L在課題實(shí)施過程中，得到了趙學(xué)軍師兄的指導(dǎo)；在論文研究過程中吳慧慧同學(xué)、來金良同學(xué)一起參與，一起討論試驗(yàn)得失，從他們那里得到許多啟迪；實(shí)驗(yàn)室的所有成員和我的室友在生活和試驗(yàn)中給了我奠大的幫助，在此一并表示感謝。本論文得到國(guó)家“十五”奶業(yè)重大科技專項(xiàng)課題和浙江省自然科學(xué)基金經(jīng)費(fèi)的資助，是研究得以開展的基礎(chǔ)，在此向有關(guān)部門和領(lǐng)導(dǎo)表示感謝十幾年求知路上，無數(shù)人相助。在此，向所有關(guān)心、幫助過我的朋友致以衷心的謝意，向悉心培育過我的老師致以崇高的敬意，向歷盡千辛萬苦撫育我成人、供我讀書的父母表示最誠(chéng)摯的謝意，父母的養(yǎng)育之恩我終生難忘桶金勇于杭州浙江大學(xué)華家池2006年S月

簡(jiǎn)介：寧夏大學(xué)碩士學(xué)位論文藥用植物多糖促進(jìn)奶牛乳腺免疫機(jī)能的研究姓名王曉峰申請(qǐng)學(xué)位級(jí)別碩士專業(yè)臨床獸醫(yī)學(xué)指導(dǎo)教師史遠(yuǎn)剛20080401ABSTRACTACHYRANTHESBIDENTATAPOLYSACCHARIDESANDLYCIUMBARBARUMPOLYSACCHARIDESASWELLASTHEEXTRACTOFPROPOLISWEREOBTAINEDINLABORATORY,WHICHWEREEMPLOYEDFORCARRYINGOUTTHEEXPERIMENTSOFPHARMACODYNAMICS，PHARMACOKINETICSANDCLINICTODEMONSTRATETHEIREFFECTSONIMPROVEMENTOFMAMMARYIMMUNOFUNCTIONINDAIRYCOW．THERESULTSSHOWEDASFOLLOWS1．THEEXTRACTINGTECHNOLOGYOFMICROWAVEFORABPANDLBPWASGRADEDUPWITHORTHOGONALTEST．THEOPTIMALPARAMETERSWEREASFOLLOWINGTREATINGTIME24MINAND16MIN，MATERIAL／SOLVENTRATIO120AND115，THEFIREPOWERGRADE9AND9；THEREWERE5AND4KINDSOFFRACTIONSINABPANDLBP．THEAVERAGERECOVERYRATEOFABPANDLBPWERE98．08％AND96．50％．THESTABILITYOFTHEABPANDLBPWEREMEASUREDWELLIN8HBYHYDROXYLBENZENEVITRI01．THEVALUEOFTHEMAXIMUMUVABSORPTIONPEAKOFTHEABPANDLBPWERE288MNAND290NM，THEABSORPTIONPEAL【OFTHEMWEREN’TAPPEAREDBETWEEN260NMAND280NMWHICHSHOWEDTHATNUCLDCACIDSANDPROTEINSWEREN’TCONTAINEDINTHETWOPOLYSACCHARIDES．THERESULTSOFINFI鼉REDSPECTROSCOPYANALYSISSHOWEDTHATFRUCTOSEANDGLUCOSEMAYBECONTAINEDINABPANDGLUCOSE，RHAMNOSEANDMANNOSEMAYBECONTAINEDINLBP．THERESULTSOFPOLARIMETRYANALYSISSHOWEDTHATTHEVALUEOFTHESPECIFICROTATIONWE托一6．70AND一150．0。AT20℃；THERESULTSOFHPLCANALYSISSHOWEDTHATTHEREEXISTED5AND6KINDSOFMONOSACCHARIDESINABPANDLBP．2．THEMEDIANLETHALDOSELD50OFABP、LBPANDCOMPOSITEPOLYSACCHARIDEWERE19．05，20．42AND4．71GK91INTHEACUTETOXICITYEXPERIMENTANDLDSL59ACG,WHICHPROVEDTHATTHETHREEACTIVEINDIGRENTSWERENONTOXICSUBSTANCES．3．THEPHARMACOKINETICSEXPERIMENTFORABPINRABBITWASINLINEWITHTHETWOCHAMBERSMODELANDITWASDISTRIBUTEDVERYQUICKLY，BUTELIMINATEDSLOWLYINBODY．4．THERESULTSOFTHELYMPHOCYTEPROLIFERATIONEXPERIMENTINDICATEDTHATTHEVALUEOFODWASHIGHERTHANTHATTHECONTROLGROUPFP0．05FORABPANDLBPANDTHEVALUEOFODOFTHECOMPLEXGROUPWASSIGNIFICANTLYHIGHERTHANTHATOFCONTROLGROUPP0．O1，THEVALUEOFODOFTHECYPOLYSACCHARIDEGROUPWASSIGNIFICANTLYHIGHERTHANTHECYGROUPFP0．01．THERESULTSOFTHEMULTIPLECOMPARISONSHOWEDTHATTHEFOURGROUPSEXISTEDSIGNIFICANTDIFFERENCES．5．IIL、，ITROBACTERIESTASISOFTHEPOLYSACCHARIDESSHOWEDTHATTHEANTIBACTERIALCIRCLEDIAMETEROFABPGROUP、LBPGROUP、EPGROUP、ABPLBPEPGROUPANDLBPRUYANKANGGROUPWERELARGERTHANTHEPOSITIVECONTROLGROUPANDNEGATIVECONTROLGROUPPO．05INDIFFERENTCONCENTRATION,THEANTIBACTERIALCIRCLEDIAMETER、ⅣEREINCREASEDWIMTHEINCREASINGCONCENTRATIONANDTHEINHIBITORYEFFECTWASALSOSTRENGLHENEDINEACHGROUP．6．THECLINICALEFFECTOFPOLYSACCHARIDESCOMPOUNDINDICATEDTHAT1HLDIFFERENTDRUGDOSEGROUP，THEGROUPOF500RAGPERINDIVIDUALWASOBVIOUSDIFFERENCECOMPARED、IRITHTHECONTROLGROUPP0．01，SCCDECREASEDASDAYSWENTBY．3INDIFFERENTADMINISTRATIONMETHOD,THEEFFECTOFTHEADMINISTRATIONGROUPWASEXTREMELYBETTERTHANTHECONTROLGROUPPO．OI，INADDITION，THEEFFECTOFTHEGROUPWHICHMJCCTEDDRUGINTHEBOTTOMOFTHEUDDERWASALSOBETTERTHANTHECONTROLGROUPPO．05，BUTTHEⅡ

簡(jiǎn)介：南京農(nóng)業(yè)大學(xué)碩士學(xué)位論文CPGODN對(duì)兩種病原菌聯(lián)合誘導(dǎo)的山羊?qū)嶒?yàn)性乳腺炎的保護(hù)研究姓名荊強(qiáng)申請(qǐng)學(xué)位級(jí)別碩士專業(yè)基礎(chǔ)獸醫(yī)學(xué)指導(dǎo)教師鄒思湘20080501中文摘要和CPG．ODN處理乳區(qū)乳中腫瘤壞死因子．0【TNF．0【水平同在感染后24H上升至最高，但感染后24HCPG．ODN乳區(qū)較PBS側(cè)下降10．38％P0．05．CPG．ODN能顯著提高感染前乳汁白細(xì)胞介素．2IL．2水平。灌注CPG．ODN乳區(qū)IL．1D在感染后24H上升至最高P0．01，而灌注PBS乳區(qū)則在感染后48H上升至最高P0．01，在感染后24H、48H較PBS乳區(qū)分別下降了L1．87％尸0．05、28．72％P0．01。灌注CPG．ODN側(cè)乳中NAGASE、MPO活性隨時(shí)間的變化表現(xiàn)為先上升后下降，在24H達(dá)到最大值；PBS側(cè)乳中NAGASE、MPO活性隨時(shí)間變化表現(xiàn)出相同的規(guī)律，但NAGASE峰值高于灌注CPG．ODN側(cè)，MPO峰值低于灌注CPG．ODN側(cè)；血清中補(bǔ)江．C‘先上升后下降，第24H時(shí)最大，第8H與第0H相比差異顯著儼0．05；IL．1P、IL．2均先下降后上升再下降，分別在第24H、48H時(shí)達(dá)最大，各時(shí)間點(diǎn)與第0H相比差異均不顯著；MPO活性隨時(shí)間的變化先下降后上升，24H最低，與0H相比差異極顯著尸O．01；NAGASE活性變化與MPO一致，到達(dá)最小值的時(shí)間為第48H，各時(shí)間點(diǎn)與第0H相比差異均不顯著。結(jié)果表明CPG．ODN可通過提高乳中IL．2水平、加速并促進(jìn)乳汁TNF．A、IL．1P的釋放，對(duì)縮短炎癥過程也有一定的作用，證實(shí)CPG．ODN對(duì)金葡茵和大腸桿菌聯(lián)合感染誘發(fā)的山羊乳腺炎有一定的保護(hù)作用。3CPG．ODN對(duì)實(shí)驗(yàn)性乳腺炎山羊乳中乳鐵蛋白含量及乳腺組織二維凝膠電泳譜的影響用ELISA法對(duì)實(shí)驗(yàn)中定時(shí)采集的乳樣進(jìn)行乳鐵蛋白含量測(cè)定。結(jié)果顯示，灌注CPG．ODN乳區(qū)乳鐵蛋白含量隨時(shí)間的變化先上升后下降，在第16H達(dá)到最大尸0．01，COMPAREDTO0THHOUR，灌注PBS的乳區(qū)乳鐵蛋白含量隨時(shí)間的變化也是先上升后下降，在第24H達(dá)到最大妒O．01，COMPAREDTO0THHO岫。CPGODN可能通過影響非特異性免疫在乳腺炎發(fā)生時(shí)發(fā)揮對(duì)機(jī)體的保護(hù)作用．此外，本實(shí)驗(yàn)以乳腺組織為對(duì)象，初步建立乳腺蛋白質(zhì)組研究的二維電泳TWO．DIMENSIONALELECTROPHORESIS，2DE技術(shù)，通過優(yōu)化各種實(shí)驗(yàn)參數(shù)，建立了乳腺組織蛋白質(zhì)表達(dá)譜，為其后全景式的蛋白質(zhì)鑒定奠定基礎(chǔ)。關(guān)鍵詞CPG．ODN；山羊；乳腺炎；金黃色葡萄球菌；大腸桿茵

簡(jiǎn)介：關(guān)于學(xué)位論文原創(chuàng)性和使用授權(quán)的聲明本人所呈交的學(xué)位論文，是在導(dǎo)師指導(dǎo)下，獨(dú)立進(jìn)行科學(xué)研究所取得的成果。對(duì)在論文研究期間給予指導(dǎo)、幫助和做出重要貢獻(xiàn)的個(gè)人或集體，均在文中明確說明。本聲明的法律責(zé)任由本人承擔(dān)。本人完全了解山東農(nóng)業(yè)大學(xué)有關(guān)保留和使用學(xué)位論文的規(guī)定，同意學(xué)校保留和按要求向國(guó)家有關(guān)部門或機(jī)構(gòu)送交論文紙質(zhì)本和電子版，允許論文被查閱和借閱。本人授權(quán)山東農(nóng)業(yè)大學(xué)可以將本學(xué)位論文的全部或部分內(nèi)容編入有關(guān)數(shù)據(jù)庫(kù)進(jìn)行檢索，可以采用影印、縮印或其他復(fù)制手段保存論文和匯編本學(xué)位論文，同時(shí)授權(quán)中國(guó)科學(xué)技術(shù)信息研究所將本學(xué)位論文收錄到中國(guó)學(xué)位論文全文數(shù)據(jù)庫(kù)，并向社會(huì)公眾提供信息服務(wù)。保密論文在解密后應(yīng)遵守此規(guī)定。論文作者簽名室麴焦導(dǎo)師簽名≈壘莖步日

簡(jiǎn)介：大連理工大學(xué)碩士學(xué)位論文致奶牛乳腺炎金黃色葡萄球菌特異性卵黃抗體的制備及活性研究姓名郭潔申請(qǐng)學(xué)位級(jí)別碩士專業(yè)生物化工指導(dǎo)教師徐永平20071221致奶牛乳腺炎金黃色萄萄球菌特異性卵黃抗體的制各及活性研究PREPARATIONANDINVITROSTUDYOFLGYAGAINSTDAIRYCOWMASTITISCAUSINGSTAPHYLOCOCCUSAUREUSABSTRACTDAIRYCOWMASTIFFSIS011EOFTHEMAINCALLS瞎OFECONOMICLOSSESINDAIRYCOWINDUSTRYANTIBIOTICS躺USEDWIDELYINPREVENTIONANDTREATMENTOFDAIRYCOWMASTIFFSWITHTHEPRESENTAWARENESSOFTHEPROBLEMSASSOCIATED謝TLLANTIBIOTICSRESISTANTANDITSRESIDUEITISURGENTTOFINDANALTERNATIVETOANTIBIOTICSCHICKENEGGYO伙IMMUNOGLOBULINIGYPROVIDESANINEXPENSIVEANDEFFECTIVE咄OFANTIBODIESFORTHEPASSIVEIMMUNIZATIONOFANIMALSITISAPROMISINGALTERNATIVEFORTHE缸EATMENTANDPREVENTIONOFBACTERIAINFECTIONS，ANDHASSHOWNTOBEEFFECTIVEAGAINSTANUMBEROFPATHOGENSTHEOBJECTIVEOFTHISSTUDYWASTOPRODUCEEGGYOLKINAMUNOGLOBULINIGNAGAINSTMASTITISEAUSINGSTAPHYLOCOCCUSALDRELLAUREUANDEVAL啊CCITSACTIVITYLAYINGHENSW料EIMMUNIZEDWITHFORMALINKILLED＆ARUREU8ISOLATEDFROMINFECTEDMAMMAL哆GLANDGYWASPURIFIEDFROMTHEWATERSOLUBLEFRACTIONWSFBYACOMBINADONOFSEVERALPURIFIC撕ONTECHNIQUESINCLUDINGSALTPRECIPITATIONSANDULTRAFILTMTIONTHEPURITYOFTHEIGYFINALLYPURIFIED塌9452％HIGHESTLITEROFSPECIFICINDETCM洫EDBYINDIRECTENZYMELINKEDIMMUNOSORBENTASSAYELISAWAS125600TOTLEIGYANDSPECIFICIGYCONTENTINYOUDMEDSUREDBYCOMPETITIVEELISAWAS956MG／N1LAND157MG／MLRESPECTIVELYN圯INHIBITINGACTIVITYWASDOSEDEPENDENTANDTHEGROWTHOFAUREUAGROWTHWASEVIDENTLYINHIBITEDBYSPECIFICIGY砒THECONCENTRATIONOF5MG／ML1MMANOELEEUONMICROSCOPYREVEALEDSLX地IFICIGYBOUNDTOAURE／／RESULTINGINSURFAOGSTRUCTURALALTERATIONSA／／REUPHAGOCYTOSISBYCOWPOLYMORPHONUCLEARLEUCOCYTE∞鋤ANDMAEROPHAGEMOINBLOODANDMILKWEREBOTHACCELERATEDBYSPECIFICIGYATCONCENTRATIONOF005MG／MLTHERESULTSINDICATEDSPECIFICIGYAGAINSTMASTIFFSCAUSING＆AUREUCOULDINHIBITGROWTHOFTHEBACTERIAANDENHANCEPHAGOCYTOSISOFCOW尸肋ⅣIJLBOTLLMILL【ANDBLOODTHECURRENTWORKSUGGESTSSPECIFICIGYHASPOTENTIALTOBEATHERAPEUTICTREATMENTFORDAIRYCOWSMASTIFFSKEYWORDSDAIRYCOWMASTITIS；EGGYOLKINUNUNOGLOBULINSTAPHYLOCOCCUSAUREUS～I(xiàn)I

簡(jiǎn)介：揚(yáng)州大學(xué)碩士學(xué)位論文山羊乳腺上皮細(xì)胞系的建立與培養(yǎng)姓名邵桃玉申請(qǐng)學(xué)位級(jí)別碩士專業(yè)動(dòng)物營(yíng)養(yǎng)與飼料科學(xué)指導(dǎo)教師趙國(guó)琦20080501邵桃玉山羊乳腺上皮細(xì)胞系的建立與培養(yǎng)山羊乳腺上皮細(xì)胞系的建立與培養(yǎng)研究生邵桃玉導(dǎo)師趙國(guó)琦教授揚(yáng)州大學(xué)動(dòng)物科技學(xué)院，225009本實(shí)驗(yàn)采用組織塊和酶消化法分離山羊乳腺上皮細(xì)胞，對(duì)純化的細(xì)胞進(jìn)行單細(xì)胞克隆并鑒定，采用二次回歸正交旋轉(zhuǎn)組合試驗(yàn)設(shè)計(jì)，MTT顯色法定量探索山羊乳腺上皮細(xì)胞培養(yǎng)的最適條件，酶聯(lián)免疫吸附實(shí)驗(yàn)與聚丙烯酰胺凝膠電泳鑒定乳腺上皮細(xì)胞是否具有正常的生理功能，最終成功建立了山羊乳腺上皮細(xì)胞系。試驗(yàn)一使用組織塊和酶消化法分離山羊乳腺細(xì)胞，結(jié)果表明組織塊培養(yǎng)可以分離純化出增殖能力較強(qiáng)的上皮細(xì)胞，I型膠原酶和透明質(zhì)酸酶聯(lián)合使用可獲得較純的上皮細(xì)胞，貼壁后增殖能力較強(qiáng)，但多次傳代后細(xì)胞增殖能力有所下降。刮除法、相差消化法以及相差貼壁法三種方法聯(lián)合使用，經(jīng)過23次傳代可以得到細(xì)胞形態(tài)均一的乳腺上皮細(xì)胞。試驗(yàn)二本實(shí)驗(yàn)進(jìn)行乳腺上皮細(xì)胞單細(xì)胞克隆，發(fā)現(xiàn)口吸管法、克隆環(huán)法和細(xì)胞稀釋法細(xì)胞克隆率分別為O、5．9％和3．5％，一共得到3株細(xì)胞克隆，形態(tài)學(xué)觀察與細(xì)胞角蛋白18鑒定為乳腺腺泡上皮細(xì)胞?？寺…h(huán)法細(xì)胞克隆率最高，但是操作很難熟練進(jìn)行，易導(dǎo)致克隆細(xì)胞的不純。細(xì)胞稀釋法操作簡(jiǎn)單易行，對(duì)細(xì)胞傷害小，克隆率較高。試驗(yàn)三采用五因素五水平的二次回歸正交旋轉(zhuǎn)組合試驗(yàn)設(shè)計(jì)，MTT顯色法定量研究不同濃度的胎牛血清FBS、氫化可的松、胰島素、轉(zhuǎn)鐵蛋白、表皮生長(zhǎng)因子EGF對(duì)山羊乳腺上皮細(xì)胞生長(zhǎng)的影響，研究發(fā)現(xiàn)山羊乳腺上皮細(xì)胞培養(yǎng)的最適條件為12％胎牛血清、4PG，ML氫化可的松、10峙／ML胰島素、8“∥M1轉(zhuǎn)鐵蛋白及6I咖NL表皮生長(zhǎng)因子的DMEM／F12。試驗(yàn)四使用催乳素誘導(dǎo)種在鋪有膠原的培養(yǎng)板中的山羊乳腺上皮細(xì)胞，采用ELISA法檢測(cè)細(xì)胞上清液中P一酪蛋白的表達(dá)檢測(cè)，培養(yǎng)48HB一酪蛋白含量為0．5MG／ML，72H為O．75MG／ML。聚丙烯酰胺凝膠電泳進(jìn)一步證明B一酪蛋白的分泌。說明本實(shí)驗(yàn)所獲得的山羊乳腺上皮細(xì)胞具有正常的生理功能。關(guān)鍵詞乳腺上皮細(xì)胞，培養(yǎng)，單細(xì)胞克隆，酪蛋白

簡(jiǎn)介：分類號(hào)密級(jí)單位代碼研究生學(xué)號(hào)LILLLLIILLILLLRLLLLI1Y17142551033510217016論文評(píng)閱人五位專家隱名評(píng)審答辯委員會(huì)主席答辯委員會(huì)委員許梓榮教授朱偉云教授張才喬教授吳躍明教授劉建新教授論文答辯日期二OO七年九月八日浙江大學(xué)中國(guó)杭州致謝值此博士學(xué)位論文完成之際，謹(jǐn)向?qū)焺⒔ㄐ陆淌谥乱宰钫\(chéng)摯的謝意本研究構(gòu)思和實(shí)施的每一個(gè)環(huán)節(jié)都凝聚了導(dǎo)師大量的心血，論文最后能順利完成更離不開導(dǎo)師的悉心指導(dǎo)。導(dǎo)師淵博的知識(shí)、嚴(yán)謹(jǐn)?shù)闹螌W(xué)態(tài)度、忘我的工作熱情和勇于探索的科學(xué)精神深深影響著我。感謝本課題組吳躍明和葉均安老師在我學(xué)習(xí)、生活和科研上給予的指導(dǎo)、關(guān)心和幫助感謝張才喬、陳安國(guó)、徐英男等老師在試驗(yàn)設(shè)計(jì)、試驗(yàn)操作過程中給我的無私指導(dǎo)和幫助感謝方維煥、余旭平、童富淡、鐘伯雄老師在試驗(yàn)技術(shù)上的無私指導(dǎo)和幫助感謝核農(nóng)所華躍進(jìn)老師在試驗(yàn)過程中提供的幫助和關(guān)心感謝富陽(yáng)市屠宰場(chǎng)在取樣過程中給予的支持和幫助感謝趙學(xué)軍、汪海峰、劉紅云、王佳墊、李菊娣等師兄師姐，廖曉霞、趙占宇、吳文旋、徐國(guó)忠、閏偉杰、孫攀峰等同學(xué)以及來金良、楊金勇、王猻、趙珂、周苗苗等師弟師妹在我試驗(yàn)和論文撰寫過程中給予的無私幫助感謝實(shí)驗(yàn)室所有成員在生活中給我的關(guān)心和幫助感謝鄔棟棟、李建營(yíng)等同學(xué)在試驗(yàn)操作過程中給予的指導(dǎo)和幫助感謝98華四齋227的黃凌霞、鄭聰霞、沈琴芳、吳媛嬡、趙麗華、黃巧芳在生活上給予的照顧、鼓勵(lì)和支持。感謝我的父母、家人和朋友在我二十年求學(xué)歷程中對(duì)我在精神方面和物質(zhì)方面的支持，是你們的理解和鼓勵(lì)，才使我有信心、并全身心地投入學(xué)術(shù)研究之中。謹(jǐn)以本文獻(xiàn)給你們感謝動(dòng)物科學(xué)學(xué)院的領(lǐng)導(dǎo)和老師對(duì)我的培養(yǎng)和關(guān)心向參加本論文評(píng)閱和答辯的全體專家致以誠(chéng)摯的謝意本論文得到浙江省自然科學(xué)基金Z305036的資助，特此表示感謝吳慧慧大忌忌于杭州浙江大學(xué)華家池2007年7月

簡(jiǎn)介：英文縮略表Ⅲ9Ⅲ3ⅢⅢⅢ1舢7Ⅲ8脅7㈣2唧Y目錄中文摘要?????????????????????????????????．IABSTRACT．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．III1前言????????????????????????????????????．．11．1奶牛乳腺發(fā)育和泌乳研究進(jìn)展??????????????????????．11．1．1奶牛乳腺發(fā)育過程???????????????????????????11．1．2泌乳周期內(nèi)奶牛乳腺細(xì)胞的增殖和凋亡?????????????????．．11．1．3奶牛乳腺上皮細(xì)胞系模型的建立?????????????????????21．1．4奶牛乳腺發(fā)育及泌乳的調(diào)控機(jī)制研究??????????????????。41．25HT對(duì)奶牛乳腺發(fā)育及泌乳的調(diào)控及機(jī)制研究進(jìn)展?????????????81．2．1乳腺內(nèi)5HT的合成與代謝???????????????????????81．2．2乳腺內(nèi)5HT再攝取?????????????????????????．．101．2．3乳腺內(nèi)5HT能系統(tǒng)?????????????????????????．．101．2．45HT對(duì)乳腺退化的調(diào)控????????????????????????111．2．55HT對(duì)乳產(chǎn)量及乳蛋白合成的調(diào)控???????????????????121．2．65HT調(diào)控泌乳和乳腺退化的機(jī)制????????????????????121．3本課題研究目的????????????????????????????132材料和方法??????????????????????????????．．132．1試驗(yàn)材料???????????????????????????????132．1．1MAC．T細(xì)胞與奶牛乳腺組織??????????????????????132．1．2主要儀器與試劑???????????????????????????．132．1．3主要試劑的配置???????????????????????????．142．2試驗(yàn)方法???????????????????????????????162．2．1體外培養(yǎng)奶牛乳腺上皮細(xì)胞系MAC．T?????????????????162．2．2RTPCR和熒光定量PCR???????????????????????。182．2．3WESTERNBLOT印記分析?????????????????????????242．3奶牛乳腺上皮細(xì)胞MAC。T形態(tài)功能鑒定及五羥色胺受體的表達(dá)?????。262．45HT對(duì)MAC．T細(xì)胞增殖和凋亡的影響及其機(jī)制初探????????????262．4．15HT對(duì)乳腺上皮細(xì)胞增殖的影響????????????????????262．4．2不同亞型的5HT受體對(duì)MAC．T細(xì)胞增殖的影響?????????????27

簡(jiǎn)介：東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文中藥透皮劑對(duì)奶牛臨床型乳腺炎的藥效學(xué)研究姓名張喜豐申請(qǐng)學(xué)位級(jí)別碩士專業(yè)基礎(chǔ)獸醫(yī)學(xué)指導(dǎo)教師張秀英20070620STUDYONPHARMACODYNAMICSACTIONOFTRADITIONALCHINESEMEDICINESPERCULANEOUSOINT僵ENTONCLINCALMASTITISOFDAIRYCATTLEABS仃ACTDAIRYCOWSMASTITISISONEOFTHEMOSTCOMMONDISEASEINCOWCULTIVATIONMASTITISNOTONLYINFLUENCEQUANTITYOFCOWMILKPRODUCTIONBUTALSOCREATESTHESERIOUSECONOMICLOSSPEOPLEHAVEPAYCLOSEATTENTIONTOPREVENTIONOFTHECOWMASTITISATPRESENTTHECOWMASTIFFSMAINPATHOGENICBACTERIAISSTAPHYLOCOCCUSAARCUS，ESCHERIEHIACOLI，STREPTOCOCCUSAGAIACTIAC，STAPH3，IOCOGCUSDYSGALACTIAEUSUALLYWEUSEANTIBIOTICTOTHERAPYTHECOWMASTITIS，HOWEVERWITHMOREANDMOREANTIBIOTICBEUSEDPEOPLEHAVEPAYATTENTIONTOTHEPROBLEMABOUTTHEREMAINOFANTIBIOTICCHINESEMEDICINALHERBDOESNOTYIELDTOLERANCEANDHAVETHENATUREOFLOWPOISONANDANTIINFLAMMATORYANDSOOILTHECOWMASTITISOFTHERAPYWITHCHINESEMEDICINALHERBISFOLLOWEDINTERESTBYMANYSEHOLAFN他RESULTSOFCLINICALPHARMACODYNAMICSEXPERIMENTSHOWEDTHATTHEOINTMENTHADGOODTHERAPEUTICACTIONONCLINICALMASTITISOFDAIRYCATTLECURINGANDEFFECTIVEPRETENTRESULTOFCYTOKINESTESTWEFOUNDTHATTHECONCENTRATIONSOFIL6TNFMINSAMPLESOFSERAANDWHEYFROMCOWSWITHCLINICALMASTITISANDNORMALHEALTHYLACTATINGCOWSEFFEETIONOFIL6、TNFMINCOWBYTRADITIONALCHINESEMEDICINESPERCUTANEOUSOINTMENT，THECONCENTRATIONSOFIL61NFNINCUREDGROUPMUCHLOWERTHANTHATINPOSITIVECONTROLGROUPRESULTOFSOD，GSHPX，NO，MDATESTTHEACTIVITIESOFSODANDGSHLAXINCLINICALMASTITISGROUPSDECLINEDSIGNIFICANTLY；WHILETHECONCENTRATIONOFNOMDAINCREASEDSIGNIFICANTLYWEFOUNDTHATEFFECTIONOFSODGSHPXINCOWBYTRADITIONALCHINESEMEDICINESPERCNTANEOUSOINTMENTTHECONCENTRATIONSOFSODGSHPXINCUREDGROUPMUCHHIGHERTHANTHATINPOSITIVECONTROLGROUPML1ETHECONCENTRATIONSOFNO，∞AINCUREDGROUPMUCHLOWERTHANTHATINPOSITIVECONTROLGROUPTHESTUDIESSHEWEDTHATTRADITIONIALCHINESEMEDICINESPERCUTANEOUSOINTMENTCALLEFFECTUALLYSTRENGTHENACTIVITYOFSOD，GSHPXDEGRADECONCENTRATIONOFFREERADICALNO’MDAANDINFLAMMATORYFACTORIL6，TNF口INSEREANDWHEYFROMCOWSWITHCLINICALMASTITISTOACHIEVEOBJEEFIVECUREDBOVINEMASTITISBYTRADITIONALCHINESEMEDICINESDERCUTANEOUSOINTMENTKEYWORDSBOVINEMASTITISTRACEELEMENTPHARMACODYNAMICSEYTOKINEANTIOXIDASEOXYGENFREERADICALM

簡(jiǎn)介：CLASSIFIEDINDEXCONFIDENTIALYES／NONOCODE10224NO120910816DISSERTATIONFORTHEMASTERDEGREEREGULATIONOFMIR1423PTOLACTATIONOFDAIRYCOWMAMMARYEPITHELIALCELLSCANDIDATEYU1EISUPERVISORPROFESSORLIQINGZHANGDEGREECATEGORYMASTEROFSCIENCECOLLEGECOLLEGEOFLIFESCIENCEFIRSTLEVELDISCIPLINEBIOLOGYSECONDLEVELDISCIPLINEBIOCHEMISTRYANDMOLECULARBIOLOGYHARBINCHINAJUNE2015東北農(nóng)業(yè)大學(xué)理學(xué)碩士學(xué)位論文355MIR1423PMIMICS轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞細(xì)胞增殖的影響26356MIR1423PMIMICS轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞凋亡的影響27357MIR1423PMIMICS轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞甘油三酯分泌的影響28358MIR1423PMIMICS轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞乳糖分泌的影響3036MIR1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞的影響30361MIR1423PINHIBOTOR轉(zhuǎn)染后奶牛乳腺上皮細(xì)胞中MIR1423P、PRLR基因的表達(dá)變化，，3362MIR1423PINHIBITOR轉(zhuǎn)染后PRLR在奶牛乳腺上皮細(xì)胞中的表達(dá)變化一31363MIR1423PINHIBITOR轉(zhuǎn)染后奶牛乳腺上皮細(xì)胞中PRLR蛋白及泌乳通路蛋白的表達(dá)變化32364MIR1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞細(xì)胞周期的影響～33365MIR1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞細(xì)胞增殖的影響一3‘366MIR1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞凋亡的影響35367MIR1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞甘油三酯分泌的影啊37368MIR一1423PINHIBITOR轉(zhuǎn)染后對(duì)奶牛乳腺上皮細(xì)胞乳糖分泌的影響384討論3941MIR1423P在不同乳品質(zhì)奶牛乳腺組織中的表達(dá)一3942MIR1423P靶基因的預(yù)測(cè)和驗(yàn)證3943MIR1423P對(duì)奶牛乳腺上皮細(xì)胞泌乳功能及相關(guān)通路蛋白的調(diào)節(jié)作用4044MIR1423P對(duì)奶牛乳腺上皮細(xì)胞增殖、細(xì)胞周期和凋亡的影響4L5結(jié)論42致謝43參考文獻(xiàn)44附錄44攻讀碩士學(xué)位期間發(fā)表的學(xué)術(shù)論文一47

簡(jiǎn)介：基金項(xiàng)目國(guó)家自然科學(xué)基金31272407；31372286；31472067SUPPORTEDBYTHENSFCNATURALSCIENCEFOUNDATIONOFCHINA31272407；31372286；31472067揚(yáng)州大學(xué)碩士學(xué)位論文路、B細(xì)胞受體信號(hào)轉(zhuǎn)導(dǎo)通路、鈣信號(hào)通路、TOLL樣受體信號(hào)通路、自然殺傷細(xì)胞介導(dǎo)的細(xì)胞毒作用、血管內(nèi)皮生長(zhǎng)因子信號(hào)轉(zhuǎn)導(dǎo)通路、白細(xì)胞跨內(nèi)皮遷移、造血細(xì)胞譜系、腸道免疫網(wǎng)絡(luò)的IGA生產(chǎn)、趨化因子信號(hào)通路、磷脂酰肌醇信號(hào)系統(tǒng)、細(xì)胞質(zhì)DNA的檢測(cè)途徑、FCYR介導(dǎo)的吞噬作用、補(bǔ)體及凝血級(jí)聯(lián)反應(yīng)、T細(xì)胞受體信號(hào)轉(zhuǎn)導(dǎo)途徑、RIGI樣受體信號(hào)通路、NOD樣受體信號(hào)通路、抗原加工和呈遞。進(jìn)一步對(duì)這些PATHWAY進(jìn)行網(wǎng)絡(luò)圖分析，發(fā)現(xiàn)了12個(gè)樞紐蛋白預(yù)測(cè)血小板堿性蛋白、間Q胰蛋白酶抑制物重鏈H3前體IHIH3、熱休克蛋白BIHSPBL、熱休克70KDA蛋白1AHSPALA、核因子NFKBP105亞基NFKBL、脂多糖結(jié)合蛋I芻LBP、RAS相關(guān)C3肉毒素底2前體RAC2、預(yù)測(cè)COPINE3亞型X1、ISOCHCRISMATASE結(jié)構(gòu)域蛋白IISOCL、未鑒定的蛋白LOE524810前體LOC524810、整合素AM前體ITGAM、整合素B2前體ITGB2?？梢猿醪綄⑦@些關(guān)鍵蛋白作為乳腺炎抗性選擇的候選蛋白進(jìn)行深入的研究。關(guān)鍵字金黃色葡萄球菌；中國(guó)荷斯坦奶牛；乳腺炎；ITRAQ技術(shù)；差異表達(dá)蛋白

簡(jiǎn)介：內(nèi)蒙古農(nóng)業(yè)大學(xué)碩士學(xué)位論文奶牛乳腺炎金黃色葡萄球菌凝聚因子AA區(qū)基因的克隆表達(dá)姓名姜曉娟申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師郝永清20080501CLONINGANDEXPRESSIONOFTHEREGIONAGENEOFCLUMPINGFACTORAOFSTAPHYLOCOCCUSAUREUSFROMBOVINEMASTITISABSTRACTMASTITISISACONUNONANDFIEQUENTLYOCCURRINGDISEASEINTHEBOVINE，ITISCAUSEDBYPATHOGENICMICROORGANISMSTHEREAREMORETHANEIGHTYPATHOGENICMICROORGANISMSWHICHCANCAUSEMASTITISINNATURE，SUCHASBACTERIUM，MYCOPLASMAFUNGIANDVIRUSESSTAPHYLOCOCCUSAUREUSISTHEMAINPATHOGENINTHEBACTERIUM，ANDTHEREARENOEFFECTIVEMEASURESTOPREVENTMASTITISCAUSEDBYSTAPHYLOCOCCUSAUREUSSOTHECLUMPINGFACTORAAREGIONGENEWERECLONEDANDEXPRESSEDINTHISARTICLETOPROVIDEMATERIASFORPRODUCEINGVACCINESOFMASTITISCAUSEDBYSTAPHYLOCOCCUSAUREUSACCORDINGTOTHECLFAASEQUENCEPUBLISHEDINGENEBANKOFNEWMANSTRAIN，PRIMERSWEREDESIGNED，THENFOURMAJORGENOTYPESOFSTAPHYLOCOCCUSAUREUSFROMHOHHOTWEREAMPLIFIEDFOURCLUMPINGFACTORAELLAAREGIONGENESOFSTAPHYLOCOCCUSAUREUSWERESEQUENCEDANDCOMPAREDWITHTHESEQUENCEOFSTANDARDNEWMANINGENBANK，THERESULTSSHOWEDTHATHOMOLOGYOFGENE3，GENE9，GENE11、L，ITHNEWMANSTRAINWERERELATIVELYHIGHERTHANGENE5WITHNEWMANITWERE978％，965％，983％AND854％RESPECTIVELYMEANWHILE，HOMOLOGYAMONGGENE3，GENE9WEREHIGHERANDLOWERINGENE5WIMTHEFORMERTHREEGENOTYPESTHEEPITOPEWEREANALYZEDBYDNASTARSOFTWAREANDTHEREWEREONEMOREANTIGENSITESINGENE一3，GENE一9THANNEWMANSOGENE9WERECHOSENFOREXPRESSIONTHENTHEFRAGMENTSWEREINSERTEDINTOTHEOFIENTMIONOFTHEVECTORPET32A，ANDARECOMBINANTPLASMIDS，PETCIFAAREGION，WEREGOTLATERTHEPLASMIDSWERETRANSFORMEDIILT0THEHOSTOFBL21DE3，ANDCLFAAREGIONPROTEINWASPRODUCEDAFTERINDUCEDBYIPTGTHETARGETPROTEINWASSOLUBLEINTHECELLSOFHOSTANDEXPRESSEDPROTEINOFHIGHPURITYWEREPURIFIEDBYHISBINDCHROMATOGRAPHYATLASTRABBITSWEREIMMUNEDBYTHEPURIFIEDPROTEINEMULSIFIED、ⅣI吐LFREUND’SADJUVANT，ANDWEHADOBTAINEDEFFICIENTANTISERUMAFTERTHREETIMESOFIMMUNIZATIONITWASPROVEDTHATTHEPROTEINEXPRESSEDWASFROMCLUMPINGFACTORAAREGIONOFSTAPHYLOCOCCUSAUREUSBYELASA，HAEMAGLUTINATIONTEST誦MTUBE，PHAGOCYTOSISCONDITIONINGTESTANDANTIADHESIONOFANTIBODIESATTHESAMETIME，111ERESULTSSHOWTHATTHEANTIBODYHASSIGNIFICANTEFFECTSOFPREVENTINGSTAPHYLOCOCCUSAUREUSFROMADHESIVINGTOFIBRINOGENOFBOVINEKEYWORDSBOVINEMASTITIS，STAPHYLOCOCCUSAURCUS，CLUMPINGFACTOR彳，CLONING，EXPRESSION

簡(jiǎn)介：CLASSIFIEDINDEXCONFIDENTIALYES／NOCODE10224NO110098DISSERTATIONFORTHEDOCTORALDEGREEREGULATIONMECHANISMOFMIR29FAMILYTOLACTATIONOFDAIRYCOWMAMMARYEPITHELIALCELLSCANDIDATEBIANYANJIESUPERVISORPROFLIQINGZHANGDEGREECATEGORYDOCTOROFAGRICULTURECOLLEGECOLLEGEOFVETERINARYMEDICINEFIRSTLEVELDISCIPLINEVETERINARYMEDICINESECONDLEVELDISCIPLINANIMALBIOCHEMISTRYANDMOLECULARBIOLOGYHARBINCHINAJUNE2015東北農(nóng)業(yè)大學(xué)理學(xué)博士學(xué)位論文3結(jié)果3731MIR29S在不同乳品質(zhì)奶牛乳腺組織中的表達(dá)37311不同乳品質(zhì)奶牛乳腺組織中RNA質(zhì)量的檢測(cè)37312小RAN測(cè)序發(fā)現(xiàn)MIR29S在不同乳品質(zhì)乳腺組織中的差異表達(dá)38313QRTPCR檢測(cè)MIR29S在不同乳品質(zhì)奶牛乳腺組織中的表達(dá)3832MIR29S靶基因的預(yù)測(cè)3933DNMT3A／3B在不同乳品質(zhì)奶牛乳腺組織中的表達(dá)及定位39331DNMT3A／3B在不同乳品質(zhì)奶牛乳腺組織中MRNA的表達(dá)39332DNMT3A／3B在不同乳品質(zhì)奶牛乳腺組織中蛋白的表達(dá)及定位4034細(xì)胞培養(yǎng)及奶牛乳腺上皮細(xì)胞泌乳功能鑒定4335DNMT3A及3B在奶牛乳腺上皮細(xì)胞內(nèi)定位的觀察4536MIR29S靶基因DNMT3A及36的鑒定46361融合DNMT3A、3B3’UTR的報(bào)告基因載體的構(gòu)建46362融合DNMT3A、3B37UTR的EGFP載體的構(gòu)建47363熒光素酶活性的檢測(cè)48364綠色熒光蛋白EGFP表達(dá)的檢測(cè)4937MIR29S超表達(dá)對(duì)奶牛乳腺上皮細(xì)胞的影響50371QRTPCR檢測(cè)MIR29SMIMICS轉(zhuǎn)染DCMECS后MIR一29S的表達(dá)50372MIR一29S超表達(dá)對(duì)靶基因DNMT3A及3BMRNA表達(dá)的影響5L373MIR29S超表達(dá)對(duì)DNMT3A及3B蛋白表達(dá)的影響52374MIR一29S超表達(dá)對(duì)DCMECS基因組GDM及甲基化酶活性的影響53375MIR29S超表達(dá)對(duì)DCMECS泌乳相關(guān)信號(hào)通路基因表達(dá)的影響54376MIR一29S超表達(dá)對(duì)DCMECS泌乳相關(guān)信號(hào)通路蛋白表達(dá)的影響54377MIR29S超表達(dá)對(duì)DCMECS活性及增殖能力的影響55378MIR一29S超表達(dá)對(duì)B一酪蛋白表達(dá)和分泌的影響57379MIR一29S超表達(dá)對(duì)細(xì)胞甘油三酯分泌的影響593710MIR29S超表達(dá)對(duì)細(xì)胞乳糖分泌的影響5938MIR一29S基因沉寂對(duì)奶牛乳腺上皮細(xì)胞的影響60381QRTPCR檢測(cè)MIR29S抑制子轉(zhuǎn)染DCMECS后MIR29S的表達(dá)60382MIR一29S沉寂對(duì)靶基因DNMT3A及3BMRNA表達(dá)的影響60383MIR一29S沉寂對(duì)DNMT3A及3B蛋白表達(dá)的影響6L384MIR29S沉寂對(duì)DCMECS基因組GDM及甲基化酶活性的影響62385MIR29S沉寂對(duì)DCMECS泌乳相關(guān)信號(hào)通路基因表達(dá)的影響63386MIR29S沉寂對(duì)DCMECS泌乳相關(guān)信號(hào)通路蛋白表達(dá)的影響64387BSP分析MIR一29B沉寂后對(duì)泌乳相關(guān)基因啟動(dòng)子甲基化的影響653885AZADC作用DCMECS后對(duì)泌乳相關(guān)基因MRAN表達(dá)的影響74389MIR一29S沉寂對(duì)DCMECS活性及增殖能力的影響78TI

簡(jiǎn)介：CLASSIFIEDINDEXCONFIDENTIALYES／NONOCODE10224NO120910812DISSERTATIONFORTHEMASTERDEGREEREGULATIONOFMIR1423PTOLACTATIONOFDAIRYGOATMAMMARYEPITHELIALCELLSCANDIDATEWANGCHUYUESUPERVISORPROFESSORLIQINGZHANGDEGREECATEGORYMASTEROFSCIENCECOLLEGECOLLEGEOFLIFESCIENCEFIRSTLEVELDISCIPLINEBIOLOGYSECONDLEVELDISCIPLINEBIOCHEMISTRYANDMOLECULARBIOLOGYHARBINCHINAJUNE2015東北農(nóng)業(yè)大學(xué)理學(xué)碩士學(xué)位論文3．3．5MIR1423PMIMICS對(duì)奶山羊鞘腺匕皮細(xì)胞活性的影響???????????????????．243．3．6MIR1423PMIMICS對(duì)奶山羊羽腺匕皮細(xì)胞周期的影響???????????????????．243．3．7MIR1423PMIMICS對(duì)奶山羊乳腺匕皮細(xì)胞凋亡的影響?????????????????253．3．8MIR1423PMIMICS對(duì)奶山羊乳腺匕皮細(xì)胞泌乳功能的影響?_????????????263．4MIR1423P基因沉寂對(duì)奶山羊乳腺匕皮細(xì)胞的影響???????????????????283．4．1MIR_142．3P{NHIB衙對(duì)奶山羊乳腺匕皮細(xì)胞MIR1423P基岡、尸，，，基因表達(dá)的影響????283．4．2MIR1423PINHIBITOR對(duì)奶山羊乳腺E皮細(xì)胞PRLR及下游信號(hào)通路蛋白的影響????????303．4．3MIR1423PINHMITOR對(duì)奶山羊乳腺E皮細(xì)胞PRLR位置及表達(dá)的影響????????????313．4．4MIR1423P劬IB．衄R對(duì)奶山羊羽腺匕皮細(xì)胞DNA增殖的影響?????????????323．4．5MIR1423PINHIBITOR對(duì)奶山羊乳腺匕皮細(xì)胞活性的影響??????????????????．333A．6MIR．1423PINHIBITOR對(duì)奶山羊乳腺匕皮細(xì)胞周期的影響????????????????333．4．7MIR1423PINHIBITOR對(duì)奶山羊乳腺E皮細(xì)胞凋亡的影I響???????????????343．4．8MIR1423PINHIBITOR對(duì)奶山羊乳腺E皮細(xì)胞泌乳叻能的影響?????????????．354‘IJ論??????????????????????????????????????????????????????????????．384．1靶基因的預(yù)測(cè)????????????????????????????????????????．384．2MIR1423P對(duì)奶山羊乳腺匕皮細(xì)胞泌乳信號(hào)相關(guān)通路蛋白的影響?????????????．．384．3MIR1423P對(duì)奶山羊鞘腺E皮細(xì)胞噌殖和1凋亡的影響?????????????????．394．4MIR1423P對(duì)奶山羊鞘腺匕皮細(xì)胞周期相關(guān)通路基因的影響??????????????????394．5MIR1423P對(duì)奶山羊乳腺E皮細(xì)胞泌乳成分的調(diào)二常???????．．．?．??．．．．．．．．．．??．?．．．．．．．．．．404．6MIR1423P對(duì)奶山羊乳腺發(fā)育乳腺發(fā)育、泌乳及退化的研究展望?????????????405結(jié)論??????????????????????????????????????????????????????????????．41JI殳I射．?????????????????????????????????????????????????????????????．．4參秀專I獻(xiàn)?????????????????????????????????????????????．43附虧己?????????????????????????????????????????????????????????????。Z19攻讀碩士學(xué)位期間發(fā)表的學(xué)術(shù)論文?????????????????????????????．5011

簡(jiǎn)介：富硒女貞子對(duì)熱應(yīng)激乳腺、卵巢組織及其細(xì)胞的影響INFLUENCEOFSEENRICHEDLIGUSTRUMLUCIDIUMFRUITONTISSUESANDCELLSOFMAMMARYANDOVARIANUNDERHEATSTRESS研究生鄒慧專業(yè)獸醫(yī)碩士導(dǎo)師許小琴教授國(guó)家自然科學(xué)基金資助項(xiàng)目NO31172354揚(yáng)州大學(xué)獸醫(yī)學(xué)院臨床獸醫(yī)學(xué)教研室2015年5月11摘要?jiǎng)e添加不同濃度的富硒女貞子水煎液、齊墩果酸、蛋白質(zhì)，檢測(cè)細(xì)胞增殖活性、細(xì)胞凋亡比例及細(xì)胞培養(yǎng)上清中SOD活性。結(jié)果表明，各濃度的富硒女貞子水煎液、齊墩果酸及蛋白質(zhì)均能顯著提高熱應(yīng)激狀態(tài)下乳腺上皮細(xì)胞的增殖活性P005、顯著降低細(xì)胞的凋亡率PO05、顯著提高細(xì)胞培養(yǎng)上清中SOD的活性P005，且富硒女貞子組明顯優(yōu)于普通女貞子組。結(jié)果提示，富硒女貞子水煎液、齊墩果酸及蛋白質(zhì)對(duì)熱應(yīng)激引起的乳腺細(xì)胞增殖活性降低及細(xì)胞凋亡有明顯的改善作用，并能提高熱應(yīng)激狀態(tài)下乳腺細(xì)胞的抗氧化功能。試驗(yàn)四富硒女貞子對(duì)熱應(yīng)激卵巢細(xì)胞生長(zhǎng)、凋亡及抗氧化的影響為探究富硒女貞予及其主要成分對(duì)高溫體外培養(yǎng)卵巢細(xì)胞增殖、凋亡及抗氧化功能的影響。按上述細(xì)胞熱應(yīng)激模型和檢測(cè)方法試驗(yàn)，結(jié)果表明，富硒女貞子水煎液及多糖能顯著提高熱應(yīng)激狀態(tài)下卵巢細(xì)胞的增殖活性JP005、顯著降低細(xì)胞的凋亡率尸005、顯著提高細(xì)胞培養(yǎng)上清中SOD的活性尸O05，且富硒女貞子組明顯優(yōu)于普通女貞子組；而富硒女貞子蛋白質(zhì)及齊墩果酸效果并不顯著。結(jié)果提示，富硒女貞子水煎液、多糖對(duì)熱應(yīng)激引起的卵巢細(xì)胞增殖活性降低及細(xì)胞凋亡有明顯的改善作用，并能提高熱應(yīng)激狀態(tài)下卵巢細(xì)胞的抗氧化功能。結(jié)論富硒女貞子水煎液對(duì)熱應(yīng)激引起的小鼠乳腺及卵巢組織發(fā)育不良有明顯的改善作用，且可以顯著提高熱應(yīng)激狀態(tài)下細(xì)胞抗氧化酶活性，改善熱應(yīng)激引起的細(xì)胞增殖活性降低及細(xì)胞凋亡。關(guān)鍵詞富硒女貞子熱應(yīng)激；乳腺細(xì)胞卵巢細(xì)胞；凋亡；抗氧化