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系統識別號 U0007-1704200714513162
論文名稱(中文) 利用LAE 技術開發抗hCG疫苗
論文名稱(英文) Development of anti-hCG vaccine by Linear Array Epitope (LAE)techology
校院名稱 臺北醫學大學
系所名稱(中) 細胞及分子生物研究所
系所名稱(英) Graduate Institute of Cell and Molecular Biology
學年度 91
學期 2
出版年 92
研究生(中文) 蘇家禛
學號 M105090010
學位類別 碩士
語文別 中文
口試日期
論文頁數 68頁
口試委員 指導教授-黃昭蓮
指導教授-施子弼
關鍵字(中) 人類絨毛膜促性腺激素
疫苗
融合蛋白抗原
醣蛋白荷爾蒙
抗體
綠膿桿菌外毒素A
線性重複排列抗原
節育
關鍵字(英) contraception
Human chorionic gonadotropin
hCG
vaccine
fusion protein antigen
glycoprotein hormone
antibody
PEIa
linaer array epitopr
LAE
學科別分類
中文摘要 人類絨毛模促性腺激素(hCG)是一種糖蛋白荷爾蒙,在授精卵形成後約10∼12天由trophoblast分泌,人類絨毛模促性腺激素的產生,在懷孕中胚胎發育的前三個月是非常必要的,因為其會刺激卵巢上的黃體產生progesterone,促使子宮內膜增厚以維持整個胚胎發育的過程。利用免疫的方法來達到節育的效果源自於1930年間,藉由引發免疫反映所產生的抗體來阻止荷爾蒙的作用,讓受孕的機制、過程中斷,無法達到正常懷孕的環境,而達到節育的效果。未達此目的,我們利用線性重複抗體辨認區段(Linear Array Epitope, LAE)之技術,合成可轉譯重複hCG部分片段之DNA序列,並構築成包含綠膿桿菌(Pseudomonas aeruginosa)外毒素A之接收器結合區及重複hCG片段的蛋白抗原。結果在動物實驗中顯示,利用此抗原所又發出的抗體的確可以達到節育70﹪的效果。
英文摘要 Human chorionic gonadotropin (hCG) is a glycoprotein hormone produced by trophoblastic cells of the placenta beginning 10 to 12 days after conception. Maintenance of the fetus in the first trimester of pregnancy requires the production of hCG, which binds to the corpus luteum of the ovary which is stimulated to produce progesterone which in turn maintains the secretory endometrium. In this study, I propose to develop immunogenes which can induce anti-hCG autoantibody, and hopefully they could be used as safe and efficient contraceptive vaccine. Toward this end, I used Linear-Array-Epitope(LAE)techniqes to construct DNA fragments encoding multicopies of hCG fragements, followed by subcloning it into a protein expression vector. The final immunogen is a fusion protein containg the receptor binding domain of Pseudomonas aeruginosa exotoxin A and multicopies of hCG fragements. After immunizing the mouse with individual immunogens, 70﹪immunized mouse lose reproductive ability. Our results reveaed that the immunogens are able to influence the reproductive ability.
論文目次 目次 中文摘要………………………………………………………….第i頁 英文摘要………………………………………………………….第ii頁 表目錄…………………………………………………………….第iii頁 圖目錄…………………………………………………………….第iv頁 第一章 緒論 第一節 人類絨毛膜促性腺激素…………………………. 第1頁 第二節 抗體產生的機轉…………………………………. 第4頁 第三節 結合毒素遞送系統與線性重複排列的方法來產生抗體 一、 綠膿桿菌外毒素A與其於疫苗的應用…………第6頁 二、 線性重複排列抗原……………………………….第8頁 第四節 anti-hCG抗體的應用 一、 節育疫苗…………………………………………第11頁 二、 癌症治療…………………………………….……第12頁 第二章 研究材料與方法 第一節 材料與試劑…………………………………………第14頁 第二節 實驗方法……………………………………………第16頁 第三章 實驗結果與分析 第一節 重複模板聚合脢連鎖反應與轉接端聚合脢連鎖反應………………………………………………….第24頁 第二節 免疫抗原的產生與純化………………………….第25頁 第三節 動物免疫實驗與抗體力價分析………………….第26頁 第四節 觀察老鼠生育能力的變化……………………… 第27頁 第五節 設計mCG序列作為抗原………………………. 第27頁 第六節 動物免疫實驗與抗體力價分析…………………. 第29頁 第七節 觀察老鼠生育能力的變化………………………. 第30頁 第四章 討論………………………………………………….. 第31頁 第五章 結論…………………………………………………... 第35頁 參考文獻………………………………………………………. 第36頁
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系統識別號 U0007-1704200714513163
論文名稱(中文) 利用LAE技術開發抗綠膿桿菌感染疫苗
論文名稱(英文) Development of Linear Array Epitope (LAE) Vaccine against Pseudomonas aeruginosa Infection
校院名稱 臺北醫學大學
系所名稱(中) 細胞及分子生物研究所
系所名稱(英) Graduate Institute of Cell and Molecular Biology
學年度 91
學期 2
出版年 92
研究生(中文) 楊雅雯
學號 M105090011
學位類別 碩士
語文別 中文
口試日期
論文頁數 72頁
口試委員 指導教授-黃昭蓮 博士
關鍵字(中) 綠膿桿菌
外膜蛋白
吞噬作用
綠膿桿菌外毒素
關鍵字(英) Pseudomonas aeruginosa
OprF
Phagocytosis
LAE
Pseudomonas exotoxin A
PE
學科別分類
中文摘要 綠膿桿菌(Pseudomonas aeruginosa)為革蘭氏陰性短桿菌,在自然界中廣泛分佈,對養分的需求少、生存條件極低,為最主要的院內感染(nosocomial infection) 伺機性感染病原菌(opportunistic infection pathogen); 但此菌極易產生抗藥性引起治療上的困難,故急須開發疫苗來預防、治療綠膿桿菌感染,以取代傳統的抗生素療法。由於綠膿桿菌會分泌多種有毒物質,其中又以綠膿桿菌外毒素A(PE)之毒性最強,會進入血流中引起菌毒血症(bacteremia),感染者的死亡率常高達70%;而根據之前的研究結果顯示,如何有效的中和阻斷PE的毒性作用,並同時能抑制菌體本身的快速繁殖及侵犯組織,是對抗綠膿桿菌感染所迫切解決的問題。有鑑於此,本論文選取綠膿桿菌的外膜蛋白F(OprF)上的抗體辨認區段(F1,F2,F3,F4,F5)利用LAE(linear array epitope)的技術合成線性重複排列的抗體辨認區段,並融合PE的domain Ia作為抗原蛋白攜帶媒介(carrier),開發出新一代兩全其美的抗綠膿桿菌感染的疫苗PEDIa-Fn(n=1,2,4,5)。目前的實驗已證明,在BALB/cByJ小鼠及英格蘭雌兔的動物實驗中,此融合蛋白PEDIa-Fn(n=1,2,4,5)所誘發出的抗體不但能專一性的識別各種不同血清型綠膿桿菌的OprF,且能與菌體表面結合,進而促進巨噬細胞對菌體的調理吞噬作用,能有效對抗綠膿桿菌的快速繁殖及其對組織的侵犯,雙管齊下,達到較佳的免疫效果,因此可預期本疫苗為一種更有效的新型抗綠膿桿菌感染疫苗。
英文摘要 Pseudomonas aeruginosa is an opportunistic pathogen that has become a major cause of nosocomial infections, and its well-known antibiotic resistance hinders therapy for P. aeruginosa infection; thus, there is considerable interest in the development of immunotherapy through either passive or active immunization. The most potent cytotoxic agent produced by P. aeruginosa is Pseudomonas exotoxin A (PE). In previous study, we know that most of P. aeruginosa infections are invasive and toxinogenic; therefore, we have designed a potent linear array epitopes (LAE) vaccine, the chimeric protein PEDIa-Fn( n=1,2,3,4,5 ), containing the receptor binding domain of PE (PE Domain Ia;PEDIa) and a linear array epitope of single motif of the outer membrane protein F (OprF) of P. aeruginosa. The potential of PEDIa-Fn(n=1,2,4,5)as a vaccine against P. aeruginosa infection is evaluated in BALB/cByJ mice and New Zealand white rabbits. At the present time, we have finished the examination of the titers of anti-PEDIa and anti-OprFFL antibodies. In addition, we have also demonstrated that the antibodies induced by chimeric protein PEDIa-F1-17 and PEDIa-F4-12 could not only recognize the OprF of various strains of P. aeruginosa but also promote the opsonophagocytic uptake of P. aeruginosa strain CCRC12902 by murine or rabbit peritoneal macrophages to prevent the colonization.
論文目次 頁次 中文摘要 ..…………………………………… 1 英文摘要 ..…………………………………… 2 緒 論 ..………………………………………..3 材料與方法 ..………………………………..12 結 果 ..……………………………………… 30 討 論 ……………………………………….. 39 參考文獻 …………………………………....46 圖 表 ……………………………………….. 51
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Dis. 130(Suppl): S94-S95 Lucy M. Mutharia, Thalia I. Nicas, and Robert E. W. Hancock (1982) Outer membrane proteins of Pseudomonas aeruginosa serotype strains. J. Infect. Dis. 146(6): 770-779 Na-Gyong Lee, Sang Bo Jung, and Wan Je Park (2000) Immunization of burn-patients with a Pseudomonas aeruginosa outer membrane protein vaccine elicits antibodies with protective efficacy. Vaccine 18: 1952-1961 Olsen B, Weinstein RA, Nathan C, and Cabins SA. (1984) Epidemiology of endemic Pseudomonas aeruginosa: why infection control efforts have failed. J. Infect. Dis. 150: 808-817 Ogata M, Chaudhary VK, Pastan I, and FitzGerald DJ. (1990) Processing of Pseudomonas exotoxin by a cellular protease results in the generating a 37,000 Da toxin fragement that is translocated to the cytosol. J. Biol. Chem. 265: 20678-20685 Pennington J. E. (1990) Pseudomonas aeruginosa. Vaccines and immunotherapy. Infect. Dis. Clin. North. Am. 4: 259-270 Rebecca S. Y. Wong, and Robert E.W. Hancock (1996) The effect of the length of a malarial epitope on its antigenicity and immunogenicity in an epitope presentation system using the Pseudomonas aeruginosa outer membrane protein OprF as the carrier. FEMS Microbiol. Letters 140: 209-214 Schaberg D. R., Culver D. H., Gaynes R. P. (1991) Major trends in the microbial etiology of nosocomial infection. Am. J. Med. 91(Suppl): S72-S75 Studier F. W., Moffatt B. A. (1986) Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned gene. J. Mol. Biol. 189: 113-130 T. Y. Chen, Chia Po Lin, and Cho Fat Hui (1999) A nontoxic Pseudomonas exotoxin A induces active immunity and passive protective antibody against Pseudomonas exotoxin A intoxication. J. Biomed. Sci. 6: 357-363 T. Y. Chen, H. F. Shang and Jaulang Hwang (1999) Recombinant protein composed of Pseudomonas exotoxin A, outer membrane proteins I and F as vaccine against P. aeruginosa infection Appl. Microbiol. Biotechnol. 52: 524-533

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系統識別號 U0007-1704200714571490
論文名稱(中文) 訶子對抗藥性金黃葡萄球菌之抗菌活性研究
論文名稱(英文) The antibacterial activity of Terminalia chebula against drug-resistant Staphylococci
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 92
學期 2
出版年 93
研究生(中文) 郭晉銓
學號 m303091004
學位類別 碩士
語文別 英文
口試日期
論文頁數 82頁
口試委員 指導教授-梁文俐 博士
關鍵字(中) 訶子
金黃色葡萄球菌
表皮葡萄球菌
MRSA
ORSA
最小抑菌濃度
chebulagic acid
1
2
3
4
6-penta-O-B-D-glucose
關鍵字(英) Chebulae Fructus
Staphylococcus aureus
Staphylococcus epidermidis
MRSA
ORSA
MIC
chebulagic acid
1
2
3
4
6-penta-O-B-D-glucose
學科別分類
中文摘要 抗生素的濫用,使抗生素抗藥性的問題變的非常嚴重,其中金黃色葡萄球菌對penicillin和methicillin的抗藥性更是深受重視。自從1980年代,PRSE (penicillin-resistant Staphylococcus epidermidis)和 MRSA (methicillin-resistant S. aureus)更變成院內重要的致病菌,PRSE和MRSA易造成嚴重的感染包括心內膜炎和致死性的休克。研究中,我們測試四十種中藥對抗PRSA (penicillin-resistant S. aureus), MRSA和PRSE, 測試抗菌的方法有瓊脂紙錠擴散法、連續稀釋法和殺菌力評估試驗。我們發現訶子的最小抑菌濃度雖然大於1mg,訶子256 ug/ml粗萃取物併用4 ug/ml oxacillin,對於MRSA(ATCC33591)具有殺菌作用。將訶子粗萃取物經有機溶媒分配之後,分別得到正己烷層, 氯仿層, 乙酸乙酯層, 正丁醇層和水層,其中正丁醇層256 ug/ml併用2 ug/ml oxacillin,對於MRSA(ATCC33591)具有殺菌作用,而1024 ug/ml氯仿層或乙酸乙酯層併用2 ug/ml penicillin G,對於PRSA(ATCC11632)也可達到殺菌效果。將正丁醇層利用管柱層析的方法,得到兩個抗菌成分chebulagic acid (CA)和1,2,3,4,6-penta-O-B-D-glucose。其中CA對MRSA(ATCC33591)的抑菌濃度為 512 ug/ml,當oxacillin併用CA256 ?ug/ml,可使oxacillin的最小抑菌濃度,由8降至0.5 ug/ml。我們測試CA對兩株MRSA臨床菌株和三株ORSA (oxacillin-resistant S. aureus)臨床菌珠的抗菌活性,其中對MRSA臨床菌株在 512 ug/ml併用0.5 ug/ml oxacillin時可達到抑菌作用;對ORSA臨床菌株則在 512 ug/ml分別併用0.5 ug/ml和2 ug/ml oxacillin時,亦達到抑菌的作用。總之,CA併用抗生素對於MRSA、PRSA、PRSE,具有良好的協同性抑菌效果。
英文摘要 Because people abuse antibiotics, bacterial resistance to antibiotics becomes very serious. Staphylococcus aureus also produce resistance to penicillin and methicillin. Since 1980s, PRSE (penicillin- resistant staphylococcus epidermidis) and MRSA (methicillin-resistant S. aureus) have become important nosocomial pathogens. PRSE and MRSA can cause serious infections, including endocarditis and toxic shock syndrome. In this study, we tested in vitro antibacterial activities of forty traditional Chinese medicines against PRSA (penicillin-resistant S. aureus), MRSA and PRSE. We tested antibacterial activities by using disc agar diffusion method, broth dilution test, and time kill assay test. Although we have found that the minimum inhibitory concentration(MIC) of Chebulae Fructus>1mg, we combined the crude extract 256 ug/ml with 4 ug/ml oxacillin, it could inhibit MRSA(ATCC33591). We isolated Chebulae Fructus by partition, and we got hexane fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction, and H2O fraction. When we combined the n-butanol fraction 256 ug/ml with 2 ug/ml oxacillin, it could inhibit MRSA(ATCC33591). We combined the chloroform fraction or the ethyl acetate fraction 1024 ug/ml with 2 ug/ml penicillin G, it could inhibit PRSA(ATCC11632). We isolated the n-butanol fraction by column chromatography, and we got two active compounds, chebulaic(CA) acid and 1,2,3,4,6-penta-O-B-D- glucose. The MIC of CA against MRSA(ATCC33591) was 512 ?ug/ml. .When oxacillin was combined with 256 ug/ml CA against MRSA (ATCC33591), the MIC of oxacillin was falling from 8 to 0.5 ug/ml. We tested the antibacterial activitiy of CA against two clinical isolates of MRSA and three clinical isolates of ORSA (oxacillin-resistant S. aureus). We combined CA 512 ug/ml with 0.5 ug/ml oxacillin, it could inhibit two clinical isolates of MRSA. We combined CA 512 ug/ml with 0.5 ug/ml and 2 ug/ml oxacillin, it could inhibit clinical isolates of ORSA. Abouve all, CA combined with antibiotics against MRSA、PRSA and PRSE has good synergical effect.
論文目次 目錄 目錄--------------------------------------------------------------------------------I 縮寫表----------------------------------------------------------------------------IV 表目錄-----------------------------------------------------------------------------V 圖目錄----------------------------------------------------------------------------VI 中文摘要-----------------------------------------------------------------------VIII 英文摘要--------------------------------------------------------------------------X 總論------------------------------------------------------------------------------XII 第一章 中藥材抗菌活性篩選 I.前言-------------------------------------------------------------------------------1 II.實驗材料 II-1.抗菌中藥材----------------------------------------------------------------4 II-2.菌種-------------------------------------------------------------------------5 II-3.儀器-------------------------------------------------------------------------6 II-4.試藥-------------------------------------------------------------------------6 III.實驗方法 III-1.藥材的萃取---------------------------------------------------------------7 III-2.檢測藥物紙錠之製備---------------------------------------------------7 III-3.菌株之解凍與活化------------------------------------------------------7 III-4.培養液及培養基之配製------------------------------------------------8 III-5.瓊脂紙錠擴散法---------------------------------------------------------8 III-6.連續稀釋法---------------------------------------------------------------9 III-7.中藥材合併臨床抗生素之最低有效抑制濃度試驗-------------10 III-8.殺菌效力評估----------------------------------------------------------10 IV.實驗結果 IV-1.瓊脂紙錠擴散法之結果----------------------------------------------12 IV-2.連續稀釋法之結果----------------------------------------------------15 IV-3.中藥材合併抗生素之結果-------------------------------------------17 V.討論--------------------------------------------------------------------------19 第二章 訶子抗菌成分分離和抗菌成分對抗藥性金黃葡萄球菌抑制活性 I.前言------------------------------------------------------------------------------21 II.實驗材料 II-1.材料-----------------------------------------------------------------------25 II-2.菌種-----------------------------------------------------------------------25 II-3.儀器-----------------------------------------------------------------------26 II-4.溶媒及試藥--------------------------------------------------------------27 II-5.管柱層析-----------------------------------------------------------------28 II-6.薄層層析-----------------------------------------------------------------28 II-7.薄層層析之展開溶媒--------------------------------------------------28 II-8.TLC檢測法--------------------------------------------------------------28 III.實驗方法 III-1.訶子的萃取-------------------------------------------------------------29 III-2.各分劃層的製備-------------------------------------------------------29 III-3.n-Butanol層的分離---------------------------------------------------30 III-4.成分結構測定----------------------------------------------------------30 III-5.有效成分對抗藥性菌株之生長曲線測定法----------------------30 III-6.有效成分合併臨床抗生素對抗藥性菌株之生長曲線測定法-31 IV.實驗結果 IV-1.各分劃層之連續稀釋法試驗----------------------------------------33 IV-2.各分劃層合併抗生素之結果----------------------------------------33 IV-3.成分結構鑑定----------------------------------------------------------36 IV-4.抽出成分的結構-------------------------------------------------------43 IV-5.chebulagic acid之抗菌活性------------------------------------------44 V.討論-----------------------------------------------------------------------------48 第三章chebulagic acid對臨床抗oxacillin與methicillin抗藥性金黃葡萄球菌抑制活性 I.前言------------------------------------------------------------------------------51 II.實驗材料 II-1.抑菌活性成分-----------------------------------------------------------53 II-2.菌種-----------------------------------------------------------------------53 II-3.儀器-----------------------------------------------------------------------54 II-4.試藥-----------------------------------------------------------------------54 III.實驗方法 III-1.chebulagic acid對抗藥性菌株之生長曲線測定法---------------55 III-2.chebulagic acid合併臨床抗生素對抗藥性菌株之生長曲線測定法------------------------------------------------------------------------55 IV.結果---------------------------------------------------------------------------57 V.討論----------------------------------------------------------------------------61 結論-------------------------------------------------------------------------------63 參考文獻-------------------------------------------------------------------------64
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系統識別號 U0007-2207201016525700
論文名稱(中文) 兩種真菌醱酵液之生物活性成分研究
論文名稱(英文) Chemical investigation of the fermented broths of Theissenia rogersii and Stilbohypoxylon elaeicola
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 98
學期 2
出版年 99
研究生(中文) 張雅雯
學號 M303097009
學位類別 碩士
語文別 中文
口試日期 2010-06-24
論文頁數 181頁
口試委員 委員-李水盛
委員-蕭哲志
指導教授-李宗徽
關鍵字(中) 真菌
誘導型一氧化氮合成酶
Theissenia rogersii
Stilbohypoxylon elaeicola
elaeicolaside
elaeicolactone
關鍵字(英) isopimarane diterpene glycoside
elaeicolaside
elaeicolactone
Theissenia rogersii
Stilbohypoxylon elaeicola
iNOS
學科別分類
中文摘要 利用抑制一氧化氮合成酶的活性分析平台來篩選真菌培養株,藉以篩檢出具有抗發炎活性的真菌株,發現某些真菌株的醱酵培養液或菌絲體萃取物,對於RAW264.7細胞的產生一氧化氮具有顯著的抑制作用。遂選定:Theissenia rogersii(#92031201)及Stilbohypoxylon elaeicola(#173)兩株進行其活性成分研究。在選擇以麥芽抽取物(malt extract)為培養基加以擴大培養後,針對醱酵液所含代謝產物進行分析與分離,計獲得12個化合物,分別為: 2,5-dihydroxymethylfuran(1)、pycnidione(2)、maltol(3)、tyrosol(4)、hymatoxin K(5)、hymatoxin L(6)、elaeicolactone(7)、5-hydroxymethylfurfural(8)、elaeicolasdie A(9)elaeicolaside B(10)、elaeicolaside C(11)和 elaeicolaside D(12),其中屬於倍半萜之化合物7,及屬於二萜糖苷的化合物9、10及12,皆為新化合物。在100 μM的濃度下,10對於RAW264.7細胞的產生一氧化氮具有顯著的抑制活性且不具細胞毒性。此外,2及10對於A549肺腺癌細胞具有強的毒殺作用,其GI50分別為5.25 nM及9.79 μM。
英文摘要 The ethyl acetate extracts of the fermented broths of Theissenia rogersii (#92031201) and Stilbohypoxylon elaeicola (#173) were found to exhibit significant activity against inducible nitric oxide synthase (iNOS) in our preliminary screening. Therefore, bioassay-guided fraction and separation of the active component from these two broths were carried out which resulted in the isolation of twelve compounds. Their structures were elucidated to be 2,5-dihydroxymethylfuran (1), pycnidione (2), maltol (3), tyrosol (4), hymatoxin K (5), hymatoxin L (6), elaeicolactone (7), 5-hydroxymethylfurfural (8), elaeicolasdie A (9), elaeicolaside B (10), elaeicolaside C (11) and elaeicolaside D (12). Of these compounds identified, 7, 9, 10 and 12, classified as respective a sesquiterpene and three diterpene glycosides, were novel chemical entities. Compound 10 exhibited significant inhibitory activity on NO production of RAW264.7 cells without any cytotoxicity. In addition, 2 and 10 exerted potent cytotoxicity against an A549 lung cancer cell line with GI50 values of 5.25 nM and 9.79 μM, respectively.
論文目次 摘要 I
Abstract II
總目錄 III
圖目錄 VII
縮寫表 XII
壹、緒論與研究目的 1
貳、炭角菌科真菌之天然物文獻回顧 5
参、實驗結果與討論 34
3.1.1 2,5-Dihydroxymethylfuran(1)之結構解析 39
3.1.2 Pycnidione(2)之結構解析 43
3.1.3 Maltol(3)之結構解析 54
3.1.4 Tyrosol(4)之結構解析 58
3.1.5 Hymatoxin K(5)之結構解析 61
3.1.6 Hymatoxin L(6)之結構解析 73
3.1.7 Elaeicolactone(7)之結構解析 82
3.1.8 5-Hydorxymethylfurfural(8)之結構解析 92
3.1.9 Elaeicolaside A(9)之結構解析 96
3.1.10 Elaeicolaside B(10)之結構解析 106
3.1.11 Elaeicolaside C(11)之結構解析 116
3.1.12 Elaeicolaside D(12)之結構解析 126
3.2 一氧化氮濃度及細胞毒性測定結果 135
3.3 A549細胞生長抑制測試結果 137
3.4 討論 138
肆、實驗部分 141
4.1 儀器設備與試劑 141
4.2 真菌材料 143
4.3 培養基配置 143
4.4.1 #92031201 培養液之成分分離與純化 145
4.4.2 #173培養液之成分分離與純化 148
4.5 單糖組成分析 152
4.6 單糖旋光度測定 153
4.7 一氧化氮(NO)濃度之測定:Griess reagent assay 154
4.8 細胞毒性測試(alamarBlue assay) 157
4.9 細胞生長測試(sulforhodamine B, SRB) 159
4.10 各成分之物理數據 161
參考文獻 165
附錄一 173
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