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全國大專校院運動代表隊學生休閒參與、涉入程度、幸福感與再參與意願影響之研究
No full text[[abstract]]本研究旨在探討全國大專校院運動代表隊學生休閒參與、涉入程度、幸福感與再參與意願間之影響關係
柔道選手歸因傾向之研究
No full text[[abstract]]近年來，運動歸因研究引起了運動心理學界的廣泛興趣，其進展已從理論研究向實際測驗的研究分析轉化。許多研究證明，個體的運動歸因傾向性，會影響著選手的情緒、態度和努力的程度
以海綿為衝擊介質呼吸性分徑採樣器佳化研究
No full text[[abstract]]作業場所呼吸性粉塵之採樣分徑標準是依據是以國際標準組織(ISO) 、歐洲標準委員會(CEN) 以及美國工業衛生技師協會(ACGIH)等三個機構所共同研商協議以50% 截取氣動直徑為4 um之微粒穿透曲線作為準則
任務導向英語寫作教學法在中華醫事科技大學之個案研究
No full text[[abstract]]近年來，技職院校越來越重視英語教育的內容與品質，而如何引導學生「想學好英文」更是許多英語教學者不斷探討的主題
Recovery of metals from Spent NiCd batteries by vitrification
No full text目錄
中文摘要 I
Abstract II
致謝 IV
目錄 V
表目錄 VII
圖目錄 VIII
第一章 前言 - 1 -
第二章 文獻回顧 - 4 -
2-1 鎳鎘電池簡介 - 4 -
2-2 鎳鎘電池組成材料 - 5 -
2-3 電池回收處理技術 - 11 -
2-4 高溫熔融法 - 12 -
第三章 研究設備與方法 - 15 -
3-1 研究流程 - 15 -
3-2 研究方法與步驟 - 18 -
3-2-1 鎳鎘電池來源及助熔劑種類 - 18 -
3-2-2 研究分析項目 - 20 -
3-2-3 高溫熱熔融操作條件試驗 - 29 -
3-2-4 排放管道中重金屬採樣 - 32 -
第四章 結果與討論 - 33 -
4-1 入料物質之特性分析 - 33 -
4-1-1 電池組成零件之特徵 - 33 -
4-1-2 鎳鎘電池各零件之金屬成分 - 41 -
4-1-3 鎳鎘電池之金屬成分 - 44 -
4-1-4 添加劑之金屬成分 - 47 -
4-2出料物質分析 - 49 -
4-2-1熔渣之成分分析 - 49 -
4-2-2金屬錠之成分分析 - 53 -
4-2-3重金屬之分離效率 - 56 -
4-3金屬宿命 - 59 -
4-3-1各出料物質之成分 - 59 -
4-4熔渣之特性分析 - 63 -
4-4-1熔渣之毒性特性溶出試驗 - 63 -
4.4.2熔渣之表面微結構分析 - 65 -
4-4-3熔渣、金屬錠之晶相分析 - 67 -
第五章 結論與建議 - 70 -
5-1結論 - 70 -
5-2建議 - 72 -
參考文獻 - 73 -[[abstract]]本研究係以高溫熱熔融法處理廢鎳鎘電池(操作溫度1450°C、持溫30分鐘)，藉由添加不同助熔劑之比例(石灰石:碎玻璃= 4:6)探討最佳助熔劑量/進料量(重量百分比)之成效、有價金屬之回收及金屬錠之分離效率，並以最佳配比進行氣相金屬採樣，瞭解其金屬元素之移動性。
研究結果顯示，熔融之產物為熔渣與金屬錠，整體性以試程1配比(電池:助熔劑= 9 : 1 )效果較佳分別為Fe(663,000 μg/g)、Ni(313,700 μg/g)、Co(27,900 μg/g)，金屬錠分離效率均可達至96%以上。金屬元素之移動率分為固相(熔渣與金屬錠)與氣相(細飛灰、粗飛灰及煙道氣)，大多數金屬均留在固相之中，少部分金屬流至氣相中分別為Cd(99%)、Hg(74%)、Pb(99%)與Zn(91%)。
熔融後之熔渣經毒性特性溶出試驗(TCLP)結果均低於法規規範之溶出標準，具有再利用潛力之資材，而有價金屬回收率各試程也可達90%以上，故高溫熱熔融法對於處理鎳鎘電池富有相當展望之潛力。
In this study, wasted Ni-Cd batteries were treated by high thermal melting method (vitrification, operating temperature 1450℃, hold 30 minutes). By using varied rate of additive melting agent(cullet:limestone =6:4) to investigate the efficiency of the optimal usage, recycle of metal and separation efficiency for ingot. Finally, under the best rate to conduct vaporized metal adopting to realize the mobility of metallic elements.
Results of this study show that melting products are slag and ingot, Procedure 1 (Battery :additive agent =9:1) got the best effect that were Fe(663,000 μg/g)、Ni(313,700 μg/g) and Co(27,900 μg/g) respectively.
The separation efficiency of ingot reached over 96%. Metal mobile rate divided into solid phase (slag and ingot) and vaporized phase (fine fly ash、cosrse ash、flue gas), large part of metal stay in solid phase and residue part of metal flow into vaporized phase that were Cd(99%)、Hg(74%)、Pb(99%) and Zn(91%).
After melting reaction, TCLP (Toxicity Characteristic Leaching Procedure) for slag is to show that testing result meet requirements of regulation standard, it means that material is reusable, metal recycle rate could reach over 90% under each process, therefore high thermal melting method (vitrification) own periscopic potential for Ni-Cd batteries treatment
The analysis of the pathogenicity relative genes in Pectobacterium chrysanthemi
Get PDF研究生中文名： 蔡孟庭
研究生外文名： Meng-Ting Tsai
論文中文名稱： Pectobacterium chrysanthemi 病原性相關基因之分析
論文外文名稱： The analysis of the pathogenicity relative genes in Pectobacterium chrysanthemi
指導教授：
中文名： 朱木貴
外文名： Mu-Kuei Chu
指導教授E-mail： [email protected]
口試委員：
中文名： 李佩芳
外文名：
中文名： 陳月茸
外文名：
口試日期： 2013-07-29
學位類別： 碩士
院校名稱： 中華醫事科技大學
系所名稱： 生物醫學研究所
畢業學年度： 102
論文出版年： 2013
學號： A00030034
語文別： 中文
論文頁數： 86
中文關鍵詞：
軟腐病菌
病原性相關基因
外文關鍵詞：
Pectobacterium chrysanthemi
個人網址名稱：
個人網址：
E-mail： [email protected]
狀態： 基本資料建檔已完成
論文中文摘要： 蝴蝶蘭是全球最受歡迎的觀賞植物也是為台灣重要外銷觀賞植物之一，近年來蝴蝶蘭已成為台灣重要的外銷花卉，其經濟栽培面積大幅擴增，但因溫室栽培環境高溫多濕，若種植過程管理不當，則易助長蝴蝶蘭病害之發生與蔓延，造成蘭園重大之損失。而蝴蝶蘭細菌性病害的主要的病原菌有三種，分別為褐斑病菌(Acidovorax avenae subsp. cattleyae)、葉斑病菌(Burkholderia gladioli)及軟腐病菌(Pectobacterium chrysanthemi)，而其中以軟腐病菌最為嚴重。本研究擬利用生物資訊學比對及分析軟腐病菌Pectobacterium chrysanthemi中第一型及第三型分泌系統中病原性相關之基因，希望能從這些篩選出可鑑別PCH及PCC差異之序列，並開發專一性引子組。本研究從NCBI Genomes資料庫中匯整Pectobacterium chrysanthemi 第一型分泌系統外膜蛋白基因與第三型分泌系統之hrp基因群序列，作為鑑別PCH及PCC分析之依據。利用生物資訊學已篩選出第一型分泌系統之eprF基因及第三型分泌系統之hrcJ、hrcT、hrcU、hrpL、hrpN及hrpX基因，都可用於開發PCR鑑定技術之用途。研究中利用分生技術已成功選殖上述hrp基因群，並將選殖株之基因序列以BLAST軟體進行比對，研究發現選殖株之序列對NCBI資料庫中PCH之hrcJ、hrcT、hrcU、hrpL、hrpN及hrpX基因的相似度分別介於88-96％、83-93％、83-92％、89-96％、85-88％及80-91％。而選殖株之序列對NCBI資料庫中PCC的相似度則較低，依序分別為73％、73％、72％、71％、74％及70％。序列間之異同可用於專一性引子組之開發，本研究已成功利用多序列比對結果設計出專一性引子組，初步測試可用於PCH及PCC的鑑別。此外對於致病相關基因有系統性的分析，發現在許多基因序列對不同來源之PCH仍有程度上的差異，是否與寄主專一性有關仍有待進一步了解。本研究中已成功選殖及表現出eprF基因，此基因的DNA長度為1389bp、分子量為52kDa，相似度為96％，確定為研究中選殖之基因，未來可大量表現蛋白質並純化，期可在抗體製作上加以進行，進一步利用免疫磁珠PCR及ELISA-PCR以增加檢測之敏感度。
論文外文摘要： Phalaenopsis became the most important products in the agriculture, the cultivation area of orchids increased rapidly. Growing the orchid in the greenhouse caused by the increasing of the temperature and humidity would yield the phytobacteria diseases, which caused enormous losses. The important phytobacteria were Acidovorax avenae subsp. cattleyae, Burkholderia gladioli and Pectobacterium chrysanthemi which caused infected diseases on orchids. One of the most serious bacterial disease agents is Pectobacterium chrysanthemi in Taiwan. This study plans to understand the pathogenicity relative genes in the type I and type III secretion system from Pectobacterium chrysanthemi. In this study, we attend to analysis the diversity between PCH and PCC, and to develop the specific primer-pairs for PCR detection. We collect the sequences of type I secretion system gene and type III secretion system genes to make the database respectively. We use the software of multiple sequences alignment to analysis the similarity among different database to understand the diversity to design specific primers. These specific primer-pairs from different genes would be used to identify PCH. Furthermore we have cloned the genes included hrcJ, hrcT, hrcU, hrpL, hrpN and hrpX genes into E. coli. The sequences of hrcJ, hrcT, hrcU, hrpL, hrpN and hrpX were used to BLAST analysis, that revealed the similarity are 88-96% ,83-93%, 83-92%,89-96%,85-88% and 80-91% to PCH,respectively. The sequences BLAST analysis to PCC revealed the similarity are 73%,73%, 72%, 71%, 74% and 70%, respectively. This study has been used the diversity of multiple sequence alignment to design specific primers. The PCR tests reveal the accuracy to PCH. This study has successfully demonstrated eprF gene, the DNA length is 1389bp, the molecular weight is 52kDa and the similarity is 96%, determine the gene for the study, overexpression of the protein and purification of antibody production Future study immuno-magnetic PCR and ELISA-PCR in order to increase the detection sensitivity.
狀態： 中外文摘要建檔已完成
論文目次： 目錄
口試委員審定書
授權書
中文摘要................................................................................................................III
英文摘要.................................................................................................................V
致謝.......................................................................................................................VII
壹、 前人研究.......................................................................................................2
一、蝴蝶蘭簡介................................................................................................2
二、蝴蝶蘭產業發展狀況................................................................................3
三、蘭花細菌性病害之種類............................................................................4
(一). 蘭花細菌性褐斑病........................................................................4
(二). 蘭花細菌性葉斑病........................................................................6
(三). 蘭花細菌性軟腐病........................................................................7
四、Pectobacterium之分泌系統..................................................................11
五、研究目的................................................................................................13
貳、 材料與方法.................................................................................................15
一、材料.......................................................................................................15
(一). 菌株來源、培養及保存.............................................................15
(二). 培養基........................................................................................15
二、染色體DNA的萃取................................................................................15
三、聚合酶連鎖反應....................................................................................16
四、PCR產物之純化....................................................................................16
五、PCR產物與質體之接合作用.................................................................17
六、核酸轉型作用........................................................................................17
七、DNA定序與引子組設計.......................................................................17
八、 PCR產物之膠體電泳分析....................................................................18
九、生物資訊分析..........................................................................................18
十、基因之蛋白質表現及純化......................................................................18
十一、蛋白質表現之分析..............................................................................19
(一). SDS-PAGE分析.........................................................................19
(二). 膠體染色...................................................................................19
(三). 西方墨點(Western Blotting).....................................................19
參、 結果.............................................................................................................21
一、第三型分泌系統hrpL基因之分析......................................................21
二、第三型分泌系統hrcT基因之分析......................................................22
三、第三型分泌系統hrcJ基因之分析......................................................22
四、第三型分泌系統hrpN基因之分析.....................................................23
五、第三型分泌系統hrcU基因之分析.....................................................24
六、第三型分泌系統hrpB基因之分析....................................................25
七、第三型分泌系統hrpX基因之鑑定......................................................25
八、利用表現載體來分析eprF基因之表現...........................................26
肆、 討論.............................................................................................................27
伍、 參考文獻.....................................................................................................29
陸、 圖表.............................................................................................................42
表1 本研究所使用菌株名稱、宿主及其分離地點............................................43
表2 本研究所使用引子組之名稱及核酸序列................................................44
表3 Pectobacterium chrysanthemi及Pectobacterium carotovorum subsp. carotovorum基因體中hrpL之相似度分析........................................................46
表4 Pectobacterium chrysanthemi及Pectobacterium carotovorum subsp. Carotovorum之染色體DNA以不同引子組所進行之PCR分析結果.............47
表5 Pectobacteriumchrysanthemi及Pectobacteriumcarotovorum subsp. Carotovorum之菌株分離來源及其註冊碼..........................................................48
圖1 Pectobacterium chrysanthemi及Pectobacterium carotovorum subsp. carotovorum基因體中hrpL之相似度分析..........................................................49
圖2 以多序列比對軟體ClustalW分析第三型分泌系統之hrpL基因序列
間之親緣關係圖....................................................................................................51
圖3 PCH菌株基因體中選殖之hrpL基因序列的BLAST分析……….....…. 52
圖4 PCH及PCC染色體以引子組(hrpL-F及hrpL-R)所進行之PCR鑑定
結果........................................................................................................................53
圖5 以多序列比對軟體ClustalW分析第三型分泌系統之hrcT基因序列
間之親緣關係圖....................................................................................................54
圖6 PCH染色體DNA以引子組(hrcT-F與hrcT-R)所進行之PCR增幅
結果........................................................................................................................55
圖7 PCH菌株基因體中選殖之hrcT基因序列的BLAST分析…...................56
圖8 PCH及PCC染色體以引子組(hrcT-F及hrcT-R)所進行之PCR鑑定
結果........................................................................................................................57
圖9 以多序列比對軟體ClustalW分析第三型分泌系統之hrcJ基因序列
間之親緣關係圖....................................................................................................58
圖10 PCH及PCC染色體以引子組(hrcJ-F與hrcJ-R)所進行之PCR鑑定
結果........................................................................................................................59
圖11 PCH菌株基因體中選殖之hrcJ基因序列的BLAST分析…………..60
圖12 PCH及PCC染色體以引子組(hrcJ-F與hrcJ-R)所進行之PCR鑑定
結果........................................................................................................................61
圖13 以多序列比對軟體ClustalW分析第三型分泌系統之hrpN基因序列
間之親緣關係圖....................................................................................................62
圖14 PCH菌株基因體中選殖之hrpN基因序列的BLAST分析……………63
圖15 PCH及PCC染色體以引子組(hrpN-F與hrpN-R)所進行之PCR鑑定結
果........................................................................................................................64
圖16 PCH及PCC染色體以引子組(hrcU-F與hrcU-R)所進行之PCR鑑定
結果........................................................................................................................65
圖17 PCH菌株基因體中選殖之hrcU基因序列的BLAST分析………….....66
圖18 PCH及PCC染色體以引子組(hrpB-F與hrpB-R)所進行之PCR鑑定
結果........................................................................................................................67
圖19 PCH菌株基因體中選殖之hrpB基因序列的BLAST分析.....................68
圖20 PCH菌株基因體中選殖之hrpX基因序列的BLAST分析……….…....69
圖21 PCH及PCC染色體以引子組(hrpX-F與hrpX-R)所進行之PCR鑑定結果............................................................................................................................70
圖22 EprF-pQE30之構築示意圖......................................................................71
圖23 EprF-pQE30蛋白質在大腸桿菌中之表現..............................................72
圖24 選殖株之蛋白在pQE30/M15表現系統中之Western blotting 分析…..73
柒、 附錄.............................................................................................................74
附錄1 第三型分泌系統之hrp相關基因群.......................................................75
附錄2 植物酚酸在第三型分泌系統之路徑......................................................76[[abstract]]蝴蝶蘭是全球最受歡迎的觀賞植物也是為台灣重要外銷觀賞植物之一，近年來蝴蝶蘭已成為台灣重要的外銷花卉，其經濟栽培面積大幅擴增，但因溫室栽培環境高溫多濕，若種植過程管理不當，則易助長蝴蝶蘭病害之發生與蔓延，造成蘭園重大之損失。而蝴蝶蘭細菌性病害的主要的病原菌有三種，分別為褐斑病菌(Acidovorax avenae subsp. cattleyae)、葉斑病菌(Burkholderia gladioli)及軟腐病菌(Pectobacterium chrysanthemi)，而其中以軟腐病菌最為嚴重。本研究擬利用生物資訊學比對及分析軟腐病菌Pectobacterium chrysanthemi中第一型及第三型分泌系統中病原性相關之基因，希望能從這些篩選出可鑑別PCH及PCC差異之序列，並開發專一性引子組。本研究從NCBI Genomes資料庫中匯整Pectobacterium chrysanthemi 第一型分泌系統外膜蛋白基因與第三型分泌系統之hrp基因群序列，作為鑑別PCH及PCC分析之依據。利用生物資訊學已篩選出第一型分泌系統之eprF基因及第三型分泌系統之hrcJ、hrcT、hrcU、hrpL、hrpN及hrpX基因，都可用於開發PCR鑑定技術之用途。研究中利用分生技術已成功選殖上述hrp基因群，並將選殖株之基因序列以BLAST軟體進行比對，研究發現選殖株之序列對NCBI資料庫中PCH之hrcJ、hrcT、hrcU、hrpL、hrpN及hrpX基因的相似度分別介於88-96％、83-93％、83-92％、89-96％、85-88％及80-91％。而選殖株之序列對NCBI資料庫中PCC的相似度則較低，依序分別為73％、73％、72％、71％、74％及70％。序列間之異同可用於專一性引子組之開發，本研究已成功利用多序列比對結果設計出專一性引子組，初步測試可用於PCH及PCC的鑑別。此外對於致病相關基因有系統性的分析，發現在許多基因序列對不同來源之PCH仍有程度上的差異，是否與寄主專一性有關仍有待進一步了解。本研究中已成功選殖及表現出eprF基因，此基因的DNA長度為1389bp、分子量為52kDa，相似度為96％，確定為研究中選殖之基因，未來可大量表現蛋白質並純化，期可在抗體製作上加以進行，進一步利用免疫磁珠PCR及ELISA-PCR以增加檢測之敏感度。
Phalaenopsis became the most important products in the agriculture, the cultivation area of orchids increased rapidly. Growing the orchid in the greenhouse caused by the increasing of the temperature and humidity would yield the phytobacteria diseases, which caused enormous losses. The important phytobacteria were Acidovorax avenae subsp. cattleyae, Burkholderia gladioli and Pectobacterium chrysanthemi which caused infected diseases on orchids. One of the most serious bacterial disease agents is Pectobacterium chrysanthemi in Taiwan. This study plans to understand the pathogenicity relative genes in the type I and type III secretion system from Pectobacterium chrysanthemi. In this study, we attend to analysis the diversity between PCH and PCC, and to develop the specific primer-pairs for PCR detection. We collect the sequences of type I secretion system gene and type III secretion system genes to make the database respectively. We use the software of multiple sequences alignment to analysis the similarity among different database to understand the diversity to design specific primers. These specific primer-pairs from different genes would be used to identify PCH. Furthermore we have cloned the genes included hrcJ, hrcT, hrcU, hrpL, hrpN and hrpX genes into E. coli. The sequences of hrcJ, hrcT, hrcU, hrpL, hrpN and hrpX were used to BLAST analysis, that revealed the similarity are 88-96% ,83-93%, 83-92%,89-96%,85-88% and 80-91% to PCH,respectively. The sequences BLAST analysis to PCC revealed the similarity are 73%,73%, 72%, 71%, 74% and 70%, respectively. This study has been used the diversity of multiple sequence alignment to design specific primers. The PCR tests reveal the accuracy to PCH. This study has successfully demonstrated eprF gene, the DNA length is 1389bp, the molecular weight is 52kDa and the similarity is 96%, determine the gene for the study, overexpression of the protein and purification of antibody production Future study immuno-magnetic PCR and ELISA-PCR in order to increase the detection sensitivity
The Effects of Diosgenin in Fibrosis Regulation of Renal Proximal Tubular Epithelial cells
Get PDF口試委員會審定書…………………………………………………..i
致謝………………………………………………………………….ii
中文摘要……………………………………………………………iii
英文摘要……………………………………………………………iv
縮寫表……………………………………………………………….v
第一章 緒論 1
一、糖尿病腎病變之盛行率 2
二、糖尿病腎臟纖維化之致病與機轉 2
三、腎臟纖維化與EMT過程之相關 5
四、高糖與糖尿病腎病變之關係 7
五、薯蕷皂素簡介 9
第二章 研究目的 10
第三章 材料與方法 12
一、試劑 13
二、方法 13
1. 細胞培養 13
( 1 ) 繼代細胞 14
( 2 ) 活化細胞 14
( 3 ) 細胞保存 15
2. 細胞數目 15
3. 細胞散射試驗.............................................................................15
4. 酵素連結免疫吸附分析 (ELISA) 16
5. 免疫螢光分析 (Immunofluorescence staining) 16
6. 細胞蛋白質總量測定 17
7. 統計分析 18
第四章 結果 19
薯蕷皂素對高糖誘導下腎近端小管細胞生長的影響................. 20
薯蕷皂素對高糖誘導下對於劑量腎近端小管型態的影響……..20
薯蕷皂素對高糖誘導下對於時間點腎近端小管型態的影響…..20
薯蕷皂素對高糖誘導下腎近端小管細胞 Fibronectin的影響…..21
薯蕷皂素抑制高糖誘導下腎近端小管細胞上皮型態的影響…..22
薯蕷皂素抑制高糖誘導下腎近端小管細胞纖維化的調控機制..22
第五章 討論與結論 23
第六章 圖表 27
第八章 參考文獻 35[[abstract]]背景：在糖尿病腎病變造成的纖維化中，高血糖是一個重要的危險因子。在先前的研究當中發現，薯蕷皂素具有抗發炎和抗癌的能力。然而，薯蕷皂素在腎纖維化的過程扮演的角色是未知的。因此，我們想要探討薯蕷皂素在腎近端小管上皮細胞高糖(D-glucose)的環境下扮演的角色。
方法：細胞培養在高糖(27.5 mM) 48小時，在最後24小時加入不同濃度的薯蕷皂素(0.1、1、10 μM)。利用免疫螢光染色法來觀察上皮－間質轉換(EMT)的標記(包括α-smooth muscle actin(α-SMA)及 E-cadherin)、訊息傳遞，及EMT的轉錄因子(Snail)。
結果：我們發現薯蕷皂素抑制高糖誘導所造成的細胞型態改變及抑制高糖誘導的細胞纖維化。並且薯蕷皂素能夠阻斷高糖誘導所升高的纖維蛋白，除此之外，薯蕷皂素能夠抑制高糖所誘導增加的α-SMA、 Snail，並且能逆轉高糖誘導減少的E-cadherin。
結論：薯蕷皂素可能抑制高糖誘導的腎近端小管纖維化可能通過調節 EMT的途徑。
BACKGROUND: Fibrosis is the key pathway of many diseases such as end-stage renal failure and liver cirrhosis. Recent study has demonstrated that transforming growth factor-β1 (TGF-β1) is the most important fibrogenesis-inducing and propagating cytokine. Diosgenin has been shown to be cancer chemopreventive and and anti-inflammatory. In this study, renal proximal tubular epithelial cells (designated as HK-2) were treated with high concentration of glucose (HG, 27.5 mM) to determine whether the Diosgenin reduces fibrosis in vitro.
METHODS: Cells were cultured in high concentration of glucose (HG, 27.5 mM) for 48 hours. Different concentrations of Diosgenin (0.1, 1, 10 μM) were added to cells for the last 24 hours. Cells were trypsinized and subjected to the following assays. ELISA was used to evaluate the secreted protein, such as fibronectin and Signal transducer EMT initiator (e.q Snail). Immunofluorescence staining was used to assay the in situ expression of proteins (e.q. fibronectin and Snail).
RESULTS: We found that the 10 μM of Diosgenin exert optimal inhibitory effects on high glucose-induced fibronectin in HK-2 cells. Diosgenin markedly inhibited HG-induced increase in α-smooth muscle actin and snail. Whereas HG-induced decrease in E-cadherin expression was reversed as well.
CONCLUSION: Diosgenin has the potential to inhibit high glucose-induced renal tubular fibrosis possibly through EMT pathway
Effects of Single Nucleotide Polymorphism of MAPK8IP1 Gene on Mood Disorders
No full text致謝 ................................................................................................................ III
中文摘要 ......................................................................................................... V
ABSTRACT ...................................................................................................... VII
目錄 ................................................................................................................ IX
第一章 前言 ................................................................................................... 1
第一節 精神疾病 ....................................................................................... 1
第二節 情感性疾患 ................................................................................... 2
第三節 臨床特徵和家族聚集在重度憂鬱症和躁鬱症 ............................. 5
第四節 精神疾病與基因 ........................................................................... 6
第五節 MAPK8IP1基因（RS1554338） ..................................................... 7
第六節 研究目的 ....................................................................................... 7
第二章 材料與方法 ....................................................................................... 8
第一節 個案 ................................................................................................ 8
第二節 測量 ................................................................................................ 8
第三節 基因分型 ...................................................................................... 10
第四節 細胞培養 ...................................................................................... 10
第五節 建構MAPK8IP1啟動子-報導基因表現質體 ................................ 11
第六節 定點突變 ...................................................................................... 11
第七節 基因轉染 ...................................................................................... 14
第八節 啟動子分析 .................................................................................. 15
第九節 統計分析 ...................................................................................... 15
第三章 結果 ................................................................................................. 17
第一節 匹茲堡(PSQI)睡眠品質問卷在不同的受試者之間有顯著差異 ... 17
第二節 疾病診斷在不同基因型間的關係 .............................................. 18
第三節 自殺與基因分型的相關性 ........................................................... 23
第四節 焦慮與基因分型的相關性 ........................................................... 25
第五節 憂鬱與基因分型的相關性 ........................................................... 27
第六節 基因分型與BMI的相關性........................................................... 28
第七節 基因分型與糖尿病(DM)無相關性 ............................................... 29
第八節 單一核苷酸基因多型性RS1554338 與MAPK8IP1基因啟動子活性有關.................................................................. 30
第四章 討論 ................................................................................................. 32
第一節 憂鬱症與雙極症盛行率 .............................................................. 32
第二節 憂鬱症和雙極症的臨床診斷 ...................................................... 32
第三節 臨床症狀與MAPK8IP1基因分型之關係 .................................... 33
第四節 睡眠品質問卷與失眠 .................................................................. 34
第五節 RS1554338對MAPK8IP1基因表現的影響 ................................. 38
第六節 RS1554338對BMI與糖尿病的影響 ........................................... 39
第五章 結論 ................................................................................................. 40
參考文獻 .................................................................................................... 41
簡歷........................................................................................................ 80
CURRICULUM VITAE ....................................................................................... 81
圖表目錄
表1-1、匹茲堡睡眠品質問卷(PSQI)分析結果............................................. 47
表1-2、PSQI各組間多重比較結果 ............................................................. 48
表2-1、疾病診斷在不同基因型間的關係(描述性統計) ............................. 50
表2-2-1、PSQI量表總分在不同基因型與診斷的關係(描述性統計) ......... 51
表2-2-2、PSQI量表總分在不同基因型與診斷的關係(推論性統計) ......... 51
表2-3-1、PSQI量表品質(QUALITY)在不同基因型與診斷的關係(描述性統計) .................................................................................................................... 52
表2-3-2、PSQI量表品質(QUALITY)在不同基因型與診斷的關係(推論性統計) ...................................................................................................................... 52
表2-4-1、PSQI量表潛伏期(LATENCY)在不同基因型與診斷的關係(描述性統計) ...................................................................................................................... 53
表2-4-2、PSQI量表潛伏期(LATENCY)在不同基因型與診斷的關係(推論性統計) ...................................................................................................................... 53
表2-5-1、PSQI量表時間(DURATION)在不同基因型與診斷的關係(描述性統計) ...................................................................................................................... 54
表2-5-2、PSQI量表時間(DURATION)在不同基因型與診斷的關係(推論性統計) ...................................................................................................................... 54
表2-6-1、PSQI量表效率(EFFICIENCY)在不同基因型與診斷的關係(描述性統計) ...................................................................................................................... 55
表2-6-2、PSQI量表效率(EFFICIENCY)在不同基因型與診斷的關係(推論性統計) ...................................................................................................................... 55
表2-7-1、PSQI量表困擾(DISTURBANCES)在不同基因型與診斷的關係(描述性統計) ............................................................................................................. 56
表2-7-2、PSQI量表困擾(DISTURBANCES)在不同基因型與診斷的關係(推論性統計) ............................................................................................................. 56
表2-8-1、PSQI量表用藥(MEDICATIONS)在不同基因型與診斷的關係(描述性統計) ................................................................................................................. 57
表2-8-2、PSQI量表用藥(MEDICATIONS)在不同基因型與診斷的關係(推論性統計) ................................................................................................................. 57
表2-9-1、PSQI量表白天功能(DYSFUNCTION)在不同基因型與診斷的關係(描述性統計).......................................................................................................... 58
表2-9-2、PSQI量表白天功能(DYSFUNCTION)在不同基因型與診斷的關係(推論性統計).......................................................................................................... 58
表3-1、自殺行為與MAPK8IP1基因型之關係 ............................................ 59
表4-1、焦慮狀態與MAPK8IP1基因型之關係 ............................................ 60
表5-1、憂鬱狀態與MAPK8IP1基因型之關係 ............................................ 61
表6-1、基因分型與BMI之關係 ................................................................. 62
表6-2、基因分型與BMI之關係(T檢定) .................................................... 62
表7-1、基因分型與糖尿病(DM)之關係 ...................................................... 63
圖一、SNP RS1554338 位於啟動子前400 BP的位置 ................................ 64
圖二、不同SNP序列對身體質量(BMI)的影響 ........................................... 65
圖三、不同SNP序列對MAPK8IP1基因啟動子活性的影響 ...................... 66
附錄
附錄一、同意計畫修正證明書 .................................................................... 67
附錄二、同意臨床試驗證明書 .................................................................... 68
附錄三、國立成功大學基因相關研究同意書 ............................................. 69
附錄四、國立成功大學醫學院附設醫院臨床試驗說明及同意書 .............. 73
附錄五、PSQI睡眠品質量表 ....................................................................... 77
附錄六、自殺、焦慮、憂鬱、躁症問卷題目 ............................................. 79[[abstract]]研究背景：情感性疾患主要是重度憂鬱症(MDD)和躁鬱症(BPD，包括BPI和BP II)，在一般人群中是常見的。然而，很少有人知道他們的臨床特點和MAPK8IP1基因的單一核苷酸多型性關係。
目的：研究MAPK8IP1基因單一核苷酸多型性與臨床特徵的關係及對MAPK8IP1基因啟動子活性的影響。
材料與方法：透過家族研究設計，共收集了1242人參加，其中包括464位個案，522個家屬和256位健康對照組，從2008至2012年在台灣南部和北部的六個地區醫院或診所收集。經由受訓過的訪員作為訪談，採用複合性國際診斷問卷訪談(CIDI)收集資料，人口統計學和臨床特徵。然後建構了啟動子上MAPK8IP1gene，伴有精神疾病或應對環境因素，並比較了不同的基因型在神經細胞上的活性變化。我們還研究了這些變化的相關性，與高壓下的精神疾病患者的表現。
結果：使用匹茲堡睡眠品質量表來評估在過去一個月的睡眠情況，在個案、家屬及健康對照組間進行比較，除睡眠時間不顯著外(P= 0.274)，其他項目都有統計上的顯著性，包括睡眠品質(P <0.001)、睡眠潛伏期(P<0.001)、睡眠效率(P <0.001)、睡眠困擾(P <0.001)、使用藥物(P <0.001)和白天功能障礙(P <0.001)等。探討基因變異與情緒障礙行為之間的關係時發現，各種情緒障礙與MAPK8IP1基因單一核苷酸多型性(SNP) rs1554338位點沒有顯著相關。但自殺行為在病患組AA/AG與GG比較，接近顯著性(P= 0.059)。探討rs1554338位點與身體質量指數(BMI)及糖尿病的關係，發現並無顯著的影響。在啟動子分析實驗上，比較rs1554338位點A與G對MAPK8IP1基因啟動子的影響，發現沒有顯著差異，為進一步探討這位點對基因表現的重要性，將之突變為T則明顯降低啟動子活性。
結論：情緒障礙與睡眠品質量表，包括睡眠品質，睡眠潛伏期，睡眠效率，睡眠困擾，藥物和白天功能障礙均有顯著性。除自殺外，情緒障礙及BMI與糖尿病和SNP MAPK8IP1基因分型無相關。但MAPK8IP1基因rs1554338位點與啟動子活性有關。
Backgrounds: Affective disorders, mainly depressive (MDD) and bipolar disorders (BPD, including BP I and BP II), are common in the general population. However, little is known about the correlation of genetic variation and clinical features.
Objectives: The objectives of this study are to correlate SNP of MAPK8IP1 gene with clinical features and to study the effects of this SNP on promoter activity.
Materials and Methods: There were 1242 participants, including 464 probands, 522 relatives and 256 controls ascertained from six regional hospitals or clinics in the Southern and Northern Taiwan from 2008 to 2012. Participants were interviewed by well-trained interviewers using Composite International Diagnostic Interview (CIDI) to collect data on demographic and clinical features. The data were collected from the interviews of patients with depression or bipolar disorder, their families and healthy people. Then we cloned the promoter of MAPK8IP1 gene, which is associated with mental diseases or responding to environmental factors, and compared the promoter activities between the promoters containing different variations. We also studied the correlation of these alleles with performance of patients suffered by mental diseases.
Results: Pittsburgh Sleep Quality Index, PSQI, was used to assess the past month sleep situation. There were significant differences among probands, relatives and controls in six items, including sleep quality (p<0.001), sleep latency (p<0.001), sleep efficiency (p<0.001), sleep disturbances (p<0.001), use of medications (p<0.001) and day-life dysfunction (p<0.001). However, sleep duration was not significant (p=0.274). We then analyze the correlation of mood disorders and SNP (rs1554338) of MAPK8IP1 gene and found that there was no significance.
However, it is more significant in suicide between MAPK8IP1 genotypes, AA/AG vs GG in patients (P=0.059). In addition, rs1554338 does not correlate to BMI or diabetics. We also found that promoter activities of MAPK8IP1 gene on A and G alleles are similar. However this site is important for MAPK8IP1 gene expression, because mutation of allele A or G to T decreased promoter activity significantly.
Conclusion: Mood disorder is associated with sleep quality, including quality, latency, efficiency, disturbances, medications and dysfunction. Except for suicide, mood disorders are not associated with SNP on MAPK8IP1 promoter. The site of MAPK8IP1 rs1554338 is important for promoter activity
Reversine inhibits cell growth and induces apoptosis and autophagy in lung cancer cells
Get PDF目錄
口試委員會審定書
授權書
誌謝
中文摘要.......................................................................I
英文摘要......................................................................II
目錄.........................................................................III
圖表目錄.......................................................................V
縮寫表...........................................................................VII
第一章、序論
第一節、Aurora kinase........................................................1
第二節、Reversine............................................................4
第三節、細胞的死亡...........................................................5
第四節、肺癌的介紹..........................................................15
第五節、研究目的............................................................18
第二章、實驗材料及方法
第一節、細胞株..............................................................19
第二節、常用儀器............................................................19
第三節、藥品試劑............................................................21
第四節、常用溶劑配製........................................................24
第五節、分析法..............................................................28
第三章、結果
第一節、Reversine對人類非小細胞肺癌細胞之存活影響...........................36
第二節、Reversine抑制非小細胞肺癌細胞癌化現象...............................36
第三節、Reversine造成人類非小細胞肺癌細胞之型態改變、並產生染色體多倍體的現象............................................................................37
第四節、Reversine造成Aurora-A及-B蛋白表現量的降低，並且影響Akt(Ser437)的磷酸化表現..........................................................................38
第五節、Reversine誘導非小細胞肺癌細胞進行凋亡作用...........................40
第六節、Reversine對Autophagy之影響..........................................41
第四章、討論..................................................................43
結論..........................................................................48
圖表..........................................................................49
附錄..........................................................................67
參考文獻......................................................................76
圖表目錄
圖1、Reversine抑制肺癌細胞的生長..............................................49
圖2、Reversine抑制肺癌細胞A549、H23、H1299癌化現象............................50
圖3、Reversine造成肺癌細胞多重細胞核及多倍體的出現............................52
圖4、Western blot分析Aurora A、Aurora B、p-Akt(Ser473)、p-GSK-3a(Ser21)及p-GSK-3b(Ser9)的蛋白質表現..........................................................55
圖5、Reversine誘導肺腺癌A549細胞進行細胞凋亡..................................58
圖6、Reversine誘導肺腺癌A549細胞進行細胞自噬作用..............................64
圖7、Reversine抑制肺癌細胞的生長並誘導細胞凋亡及細胞自噬作用之訊息傳導路徑....66
附錄一........................................................................67
附錄二........................................................................68
附錄三........................................................................69
附錄四........................................................................70
附錄五........................................................................71
附錄六........................................................................72
附錄七........................................................................73
附錄八........................................................................74
附錄九........................................................................75[[abstract]]Reversine ,2-(4-morpholinoanilino)-6-cyclohexylaminopurine 為小分子的普林合成類似物，曾被應用於幹細胞的研究上。過去有研究曾指出Reversine對於腫瘤具有抑制的效果，但它在肺癌的效果仍尚未明確。我們以Reversine處理肺癌細胞株A549、H23及H1299，並探討其相關機制路徑。首先，我們證實Reversine能抑制肺癌細胞的生長及聚落形成。進一步的研究顯示Reversine會誘導細胞週期停滯於G2/M期，並且產生細胞多倍體的現象。Reversine會降低肺癌細胞的Aurora-A及Aurora-B蛋白表現，並且降低下游p-Akt(Ser473)、p-GSK-3alpha(Ser21)及p-GSK-3beta(Ser9)的蛋白表現。 切割之caspase-3及PARP的出現證實Reversine可誘導肺癌細胞凋亡。LC3-II蛋白的增加顯示細胞自噬作用也參與其中。總結，Reversine可抑制肺癌細胞的生長及誘導細胞凋亡與細胞自噬作用，這樣的結果可以作為未來肺癌治療藥物開發上的一個新方向。
Reversine ,2-(4-morpholinoanilino)-6-cyclohexylaminopurine, a small synthetic purine analogue, has been used for stimulating stem cell dedifferentiation. It has been reported that reversine is effective in tumor suppression, but it’s effect in lung cancer cells remains unclear. We treated lung cancer cell lines A549, H23 and H1299 with reversine and disseted the related pathways. First, we demonstrated reversine inhibited cell growth and colony formation. Further results revealed reversine induced cell cycle arrested at G2/M phase and formed polyploidy. The expressions of Aurora-A, Aurora-B, p-Akt(Ser473), p-GSK-3alpha(Ser21) and p-GSK-3beta(Ser9) were down-regulated The presence of cleaved caspase-3 and PARP demonstrated reversine in reversine treated cells could induce apoptosis. That the increased LC3-II protein was present after reversine treatment implied that the autophagy is also involved. Taken together, reversine suppressed cell growth of lung cancer cells and induced apoptosis as well as autophagy, which may be an unique advantage for developing novel therapeutic agents for treatment of lung cancers in the future
Biosorption of heavy metals from aqueous solution with coffee grounds
No full text目錄
中文摘要 ............................................... -I-
英文摘要 .............................................. -II-
誌謝 ................................................. -IV-
目錄 .................................................. -V-
表目錄 .............................................. -VIII-
圖目錄 ................................................ -IX-
第一章 前言.............................................-1-
1-1 前言...............................................-1-
1-2 研究目的...........................................-2-
第二章 文獻回顧.........................................-3-
2-1何謂重金屬...........................................-3-
2-2重金屬的危害.........................................-3-
2-3常見的重金屬汙染.....................................-4-
2-3-1 鉛.................................................-4-
2-3-2 鉻.................................................-4-
2-3-3鎘.................................................-6-
2-4重金屬去除之方法.....................................-7-
2-4-1生物吸附............................................-8-
2-5 咖啡渣吸附重金屬的吸附模式..........................-8-
2-5-1 吸附現象...........................................-8-
2-5-2 吸附模式...........................................-9-
2-5-3 Freundlich 方程式等溫吸附模式.......................-13-
2-5-4 Langmuir 方程式等溫吸附模式.......................-14-
第三章 實驗材料與設備..................................-16-
3-1氣體與藥品..........................................-16-
3-1-1氣體..............................................-16-
3-1-2 藥品..............................................-16-
3-2 實驗設備與儀器.....................................-16-
第四章 研究方法........................................-18-
4-1 吸附劑的製備.......................................-19-
4-2吸附質的製備及測定..................................-19-
4-2-1 重金屬水溶液之調配................................-19-
4-2-2 吸附值pH值測定...................................-19-
4-3 實驗內容..........................................-20-
4-4吸附實驗............................................-20-
4-5 水溶液中重金屬離子濃度的量測.......................-21-
4-5-1原子吸收光譜儀....................................-21-
4-5-2感應耦合電漿原子發射光譜分析儀.....................-21-
4-5-3 重金屬離子濃度的測定步驟..........................-22-
第五章 結果與討論......................................-23-
5-1 pH影響吸附劑吸附效果...............................-24-
5-2 溫度影響吸附劑吸附效果.............................-26-
5-3 攪拌速度對吸附效果的影響...........................-28-
5-4 吸附劑量對吸附效果的影響...........................-30-
5-5 離子起始濃度對吸附效果的影響.......................-33-
5-6 Langmuir 方程式等溫吸附模式.........................-35-
5-7 Freundlich 方程式等溫吸附模式........................-37-
第六章 結論............................................-40-
參考文獻...............................................-41-[[abstract]]中文摘要
造成早期水污染的來源大多來自於大自然，而這些自然物質容易被存在於大自然的微生物分解或是自然的降解，但到了現今社會，隨著科技不斷的發展，所造成的汙染也愈加不同愈加複雜，且有大部分的汙染是無法被微生物分解或自然降解的，水汙染中的重金屬就是重要的一例，甚至經由生物累積反而危害了人類。
本研究主要目的為利用廢棄咖啡渣為吸附劑，廢棄的咖啡的優點是成本低廉，且對重金屬吸附效果佳，本文主要探討其吸附劑對重金屬最佳的操作條件，並計算出吸附反應動力學的相關係數，以確認等溫吸附模式，探討其吸附能力。由研究結果得知，咖啡渣對重金屬鉛、鎘、鉻的吸附效果佳，而對鉛的吸附效果為最佳，而在等溫吸附動力學分析中，發現Langmuir等溫吸附模式與Freundlich等溫吸附模式皆能描述咖啡渣對重金屬吸附之行為，總結咖啡渣對重金屬吸附是一有潛力的生物素材，未來可望實際應用於廢水重金屬的去除。
英文摘要
Early water pollution was make from nature, and those nature matter is easy to decomposed by microorganism or degradation by nature. As time goes by technology develop, which maked pollution have more different and complicated , in this time. Many pollution were not been decomposed by microorganism or degradation by nature. The most improtant example is heavy metal pollution which one of water pollution. That will hurt human by the way of bioaccumulation.
This research is used coffee grounds as adsorbent. It’s merit using coffee grounds of cheap, and it is good for absord to heavy metal. The main research is the best operant conditioning of adsorbent to heavy metal , and account .Correlation coefficient of adsorption dynamycs. To confirm equation and research adsorption. We were know by research ,coffee grounds is good for adsorb to lead、cadmium、chromium, in those the best one is adsorb to lead. In equation analyze to discover “Langmuir equation.”Langmuir equation and Freundich equation those two are can describe coffee goop adsorb to heavy metal. To sum up, coffee goop is a potential matter to asdsorb heavy metal, there will used to degrease heavy metal in water pollution , in the future.
Keywords：biosorption; heavy metal; dynamics; coffee ground