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Study of Potentiometric Biosensor For The Determination of Malachite Green
Get PDF口試委員審定書
授權書 ii
中文摘要 iii
英文摘要 iv
致謝 vi
第一章 緒論
1.1 研究背景 -1-
1.2 研究目的與動機 -3-
1.3 論文架構 -7-
第二章 文獻回顧及基礎原理
2.1 生物感測器
2.1.1 生物感測器簡介 -8-
2.1.2 生物感測器之分類 -9-
2.2 延伸式閘極感測場效電晶體之原理 -20-
2.3 電子材料的應用 -23-
2.4 酵素的特性 -24-
2.4.1 酵素催化過程 -24-
2.5 乙醯膽鹼酯酶生理作用機制 -25-
2.5.1 乙醯膽鹼酯酶催化受質之水解過程 -28-
2.6 孔雀綠簡介 -30-
2.6.1 孔雀綠之應用 -31-
2.6.2 孔雀綠之作用機制 -32-
2.6.3 孔雀綠之代謝機制 -33-
2.6.4 孔雀綠之毒性 -34-
2.6.5 孔雀綠對膽鹼酯酶抑制作用之文獻探討 -37-
2.6.6 世界各國對孔雀綠的管制 -38-
2.6.7 檢測孔雀綠之方法 -39-
第三章 材料方法
3.1 實驗材料 -41-
3.2 儀器與器材 -42-
3.3 實驗流程 -43-
3.4 感測器之製作 -44-
3.5 延伸式離子感測場效電晶體之pH 感測特性 -45-
3.6 感測器之酵素薄膜固定化步驟 -45-
3.7 量測系統 -46-
3.8 抑制劑之抑制試驗 -47-
第四章 結果與討論
4.1 感測器 pH 感測特性探討 -48-
4.2 乙醯膽鹼感測特性探討 -49-
4.3 孔雀綠量測方法之探討 -51-
4.4 孔雀綠感測器之反應時間 -54-
4.5 抑制百分比之參數 -55-
4.6 生物感測器與現有商業化測量探討 -60-
4.7 未來展望 -60-
第五章 結論 -61-
參考文獻 -62-[[abstract]]孔雀綠是三苯甲烷類染料，從 1930 年開始廣泛用於水產養殖業用於保護及治療在真菌及體外寄生蟲的感染。孔雀綠會殘留在魚的脂肪組織中，對於哺乳類細胞具有高度細胞毒性也是肝腫瘤促進劑，所以，孔雀綠現今已經成為極具爭議的化合物且對免疫系統、生殖系統有風險還有遺傳毒性以及致癌的可能。孔雀綠已被證明對膽鹼酯脢具有可逆的活性抑制作用，會對乙醯膽鹼酯酶上的酯基活化位置進行可逆的混合型抑制。本篇論文中，我們利用孔雀綠會抑制膽鹼酯酶的特性以電位式生物感測器進行孔雀綠感測器之開發。本論文生物感測器之架構為將乙醯膽鹼酯酶(AchE)固定於氧化銦錫膜上作為生物識別裝置，催化水解乙醯膽鹼素(Ach)伴隨著產物氫離子的衍生使膜上的 pH 值產生改變而檢測到電壓值。最佳的量測方法是利用乙醯膽鹼素泡製抑制劑，量測抑制劑之反應時間為五分鐘，乙醯膽鹼酯酶量測乙醯膽鹼素之偵測極限為 1 mM，電鰻乙醯膽鹼酯酶濃度0.73125 mg/ml 搭配乙醯膽鹼素濃度 8 mM 進行不同濃度抑制劑之量測，抑制劑濃度與抑制百分比成正比，經實驗結果顯示本論文發表之架構可量測到 1 ppm 的孔雀綠含量，利用乙醯膽鹼酯酶可被孔雀綠抑制的此種機制來發展出檢測孔雀綠生物感測器。
Malachite green (MG) is a triarylmethane dye has been used worldwide in the aquaculture industry as a fungicide for the effective prevention and treatment of fungal infections and ectoparasites since the early 1930s. MG is deposited in fatty tissue of the fish and has highly cytotoxin to mammalian cells and also acts as a liver tumor-enhancing agent, so, MG has now become a
highly controversial compound due to the risks associated, including its effects on the immune system, reproductive system and its genotoxin and carcinogenic properties. MG was shown to be a an inhibitor of acetylcholinesterase (AcChE). In this study, the characteristics of inhibitory effects were applied to development malachite green potentiometric biosensor. The AcChE was
immobilized on the sensitive structure with Indium Tin Oxide (ITO) for the biosensor that the AcChE as a biological recognition device that can catalyze hydrolyses acetylcholine (Ach), however these reactions result was generation of hydrogen ions, causing the change of pH in membrane, thus allowings use of the method for measuring voltage value. The best measurement method is the malachite green solution was prepared as a mixture of acetylcholine solution , results shows that the detection method for 1 ppm malachite green inhibition effect can decrease the hydrogen ions in the sensing membrane. The response time for detection malachite green was estimated to be about five minutes. The results indicated that the LOD for Ach detection is 1mM. When the Electrophorus electricus 0.73125 mg/ml enzyme concentration with acetylcholine concentration 8 mM measure for different concentrations
inhibitor, proportional to inhibitior concentrations and inhibition. In our experiential results shows that the structure can be a malachite green biosensor for detecting 1 ppm malachite green. Use of mechanisms such malachite green inhibited acetylcholinesterase could be the development for detect the malachite green biosensor
The Effects of Visual Training on Lens Accommodation Relaxation
Get PDF簽名頁……………………………………………………………… ii
誌謝………………………………………………………………… iii
摘要………………………………………………………………… iv
Abstract…………………………………………………………… v
目錄………………………………………………………………… vii
圖目錄……………………………………………………………… x
表目錄……………………………………………………………… xii[[abstract]]本研究探討針對18歲的大學新鮮人實施視力訓練，能否有效改善長期看近所產生的調節問題。過度調節會造成水晶體膨脹導致近視，反之，若放鬆調節，使水晶體回復不膨脹，就能控制近視，並且視力異常所帶來頭痛、視力模糊等症狀也能獲得改善。 本研究以6種測量方法(調節近點、斜位測量、正負相關調節、調節幅度、調節反應、調節靈活度) ，評估放鬆調節對改善視力的效果。實驗組給予簡單的視力訓練(交替鏡片搖擺法、遠近字母/數字卡法)，對照組則以望遠凝視的方式放鬆調節，經過14天後，比較斜位稜鏡量、調節近點、正相關調節與負相關調節之變化，發現實驗組與對照組有顯著差異p<0.05。證明短時間內使用視力訓練能有效放鬆水晶體調節，並且改善視力與減輕頭痛等症狀。 教育部自民國八十八年起，於總計五年半的時間內，推展學童做集體望遠凝視。結果顯示，民國九十三年底國小六年級近視比率，降至50%以下，比較實驗前的55%，說明望遠凝視計畫對兒童近視罹患率的改善效果不大，100人中只能減少5人，這與本人實驗結果相呼應，在配戴眼鏡的18歲學生中實施遠眺14天實驗前後差異並不明顯，可是實施視力訓練後有顯著差異。證明已配戴眼鏡者要放鬆水晶體調節，在短時間最有效果的方法是視力訓練。
This project investigated the effects of vision training for the relief of accommodation problems after long-term near vision work. Ocular accommodation leads to change of lens curvature with ensuing myopia. If the lens is relieved from excessive accommodation, myopia may be controlled and the concurrent headaches, blurred vision, and other symptoms may be alleviated. Six parameters were measured, including near point accommodation (NPA), phoria measurement, positive relative accommodateion (PRA), negative relative accommodation (NRA), accommodation amplitude, accommodative response, and accommodative facility, to assess the effects of accommodative relaxation on myopia. The simplest vision training, lens rock along with far/near number or letter card, was performed to relax the accommodation. After 14 days of intensive training, oblique prism amount, accommodation near point, positive relative accommodation (PRA), and negative relative accommodation (NRA) were significantly different between the experimental and control groups (P <0.05). The results of this study proved that a short period of visual training can be performed to relief effectively the excessive accommodative problems, so that the headache and other symptoms associated with myopia may be alleviated
The Study of Chinese Herbs Used for Sun Protection in Cosmetics
No full text中文摘要.....................................................................2
英文摘要.....................................................................3
目錄.........................................................................4
圖目錄.......................................................................8
表目錄.......................................................................10
第一章、緒論
1.1 研究動機.................................................................11
1.2 研究目的.................................................................11
第二章、研究背景
2.1 紫外線與皮膚的關係
2.1.1紫外線之介紹............................................................13
2.1.2紫外線對皮膚的傷害.......................................................18
2.1.3長期紫外線對皮膚的傷害...................................................19
2.1.4光老化現象等級..........................................................20
2.1.5皮膚組織的改變..........................................................22
2.2 皮膚的生理構造
2.2.1 皮膚的構造...........................................................23
2.2.2 皮膚的層次..........................................................................23
2.2.3 皮膚的生理功能........................................................24
2.2.4 皮膚的分類...........................................................27
2.3 防曬的定義
2.3.1 防曬能力的評估方法....................................................28
2.3.2 防曬劑的種類.........................................................31
2.3.3紫外線吸收劑具環境荷爾蒙作用............................................33
2.3.4紫外線吸收劑能引起過敏與突變............................................33
第三章、實驗大綱
3.1實驗藥品..................................................................37
3.2實驗儀器..................................................................39
3.3實驗步驟..................................................................40
3.3.1 稀乙醇抽提...........................................................40
3.3.2紫外光吸光能力(UV/Vis光譜分析).........................................41
3.3.3細胞毒性試驗(Cytotoxic Assay).........................................42
3.3.4細胞存活率試驗(MTT assay kit).........................................43
3.3.5重金屬含量試驗........................................................44
3.3.6化粧品中微生物檢驗.....................................................47
3.3.7防曬係數分析..........................................................47
第四章、結果與討論
4.1吸光能力測試(UV/Vis光譜分析)...............................................49
4.2細胞毒性(LDH)試驗及細胞存活率(MTT).........................................65
4.3重金屬含量檢測............................................................70
4.4化粧品中微生物檢測.........................................................71
4.5防曬係數分析..............................................................72
第五章、結論.................................................................73
第六章、參考資料..............................................................75[[abstract]]本研究著重在中草藥植物萃取液之防曬成份之探討與應用以應用在化粧品配方之評估，本研究選擇20種於文獻記載指出具防曬美白及抗氧化作用之中草藥，進行紫外光吸收測試，並從中篩選出松樹皮、黃芩、薑黃水萃液均具有吸收紫外光區光線之能力，然而其中又以松樹皮水萃取物吸收光譜能力表現最佳。
加入不同濃度之松樹皮萃取液進行細胞存活率(MTT)及細胞毒性試驗(LDH)，發現人類表皮細胞(HS-68) 在濃度3%之松樹皮萃取液中細胞存活率最高且細胞毒性低，將此一濃度之樣品加入乳液中進行防曬係數測試，其防曬係數可達SPF8.81。
將實驗結果實際運用在化妝品配方中，評估體外試驗結果與產品實際運用效果之差異性，結果顯示松樹皮萃取液具有作為化妝品防曬劑的潛力。
This research investigates herb extracts as a potential sun screen ingredient in cosmetics. As cited in this experiment twenty herbs were tested for their sun protection and whitening factor as well as their anti-oxidizing properties.
Pine bark, scutellaria, and turmeric extract showed viable results in ultraviolet ray absorption, in particular pine bark extract showed the most significant viability in spectrum absorption spectroscopy (spectrophotometer or spectrometer).
By testing of MTT assay kit (MTT) and Cytotoxic Assay (LDH) in different concentrations of the pine bark extract, it was found that the cells in the epidermis (HS-68) had the highest survival rate and lowest toxicity levels in 3% concentration pine bark extract. Adding the same concentration of the extract into emulsion to test out its sun protection factor consequently yielded a SPF 8.81.
The experimentation resulted in a practical application of the extract in the cosmetic formula. Assessment of in vitro tests revealed a differentiation in outcome, the results do show the pine bark extract can be used as cosmetic sunscreen
The Association Study Of Metabolic Syndrome With Genetic Polymorphism In Chronic Schizophrenic Inpatients
No full text目錄
審定書
授權書
目錄 Ⅰ
圖目錄 IV
表目錄 V
謝誌 VI
中文摘要 VIII
Abstract XI
第一章、文獻回顧 1
第一節、精神分裂症..........................................................................1
壹、精神分裂症之定義..............................................................1
貳、精神分裂症之演變..............................................................1
參、精神分裂症之流行病學......................................................4
肆、精神分裂症之常見症狀......................................................4
伍、精神分裂症之發病病程......................................................5
陸、精神分裂症之分類..............................................................6
柒、精神分裂症之病因..............................................................7
第二節、代謝症候群..........................................................................9
壹、代謝症候群之演變................................................................9
貳、代謝症候群之臨床診斷標準..............................................10
1. 國外代謝症候群之臨床診斷標準...............................10
2. 國內代謝症候群之臨床診斷標準...............................13
參、代謝症候群之核心異常......................................................14
1. 代謝症候群與肥胖.......................................................14
2. 代謝症候群與胰島素阻抗...........................................15
3. 代謝症候群與基因及其他危險因子...........................17
4. 罹患代謝症候群之高風險因素（流行病學）.............19
第三節、精神分裂症與代謝症候群之相關性探討..........................21
第四節、 ApoE、5-HT2C及PPARγ基因與代謝症候群...............22
1. Apoe基因與代謝症候群...............................................22
2. 5-HT2C基因與代謝症候群..........................................24
3. PPARγ基因與代謝症候群...........................................25
第二章、研究目的.....................................................................................29
第三章、研究方法......................................................................................30
第一節、研究對象.............................................................................30
第二節、基因檢測..............................................................................30
第三節、研究流程及方法.................................................................32
第四節、統計分析..............................................................................43
第四章、研究結果 44
第五章、討論 48
第六章、結論 52
第七章、參考文獻.................................................................................... 53
第八章、圖與表....................................................................................... 71
圖目錄
圖一、ApoE PCR結果之電泳圖......................................................... 71
圖二、5-HT2C Cys23Ser PCR結果之電泳圖......................................72
圖三、PPARγPro12Ala PCR結果之電泳圖....................................... 73
圖四、ApoE RFLP結果之電泳圖....................................................... 74
圖五、5-HT2C Cys23Ser RFLP結果之電泳圖....................................75
圖六、PPARγ Pro12Ala RFLP結果之電泳圖.................................... 76
表目錄
表一、研究人口之參數........................................................................ 77
表二、精神分裂症患者代謝症候群與基因多型性之分析................ 78
表三、精神分裂症患者代謝症候群與基因單套體之分析.................79[[abstract]]中文摘要
研究背景與目的：近幾年來，不管是一般大眾或者是精神分裂症（schizophrenia）患者，其代謝異常症狀已引起多方關注。代謝異常症狀是一群會引發心血管疾病的因子，包括肥胖、血脂異常、高血壓、高血糖等。異常結果，常會一併發生，人們把這些現象亦稱為代謝症候群（metabolic syndrome；Mets）。目前已知，代謝症候群會增加死亡率、心血管疾病及中風罹患率，且其致病機轉仍不清楚，而其盛行率一區域差異甚大暗示著基因可能是一個調控因子。載脂蛋白Apo（apolipoprotein）E是低密度脂蛋白VLDL、chylomicron remnants 及 高密度脂蛋白HDL的部分組成物。ApoE可以和apoE receptor及LDL receptor鍵結，在脂質代謝中扮演一個重要角色。一般認為過度飲食與久坐的生活模式與代謝症候群有所關聯，血清素（serotonin, 5-HT）系統和許多行為有關，其中一項就是進食；許多研究發現，抗精神病藥物引起之體重增加與5-HT2C receptor基因多型性有關，近年來也有文獻指出5-HT2C receptor 基因多型性指出與精神分裂症病人之代謝症候群有關。過氧化小體增生劑活化接受體Peroxisome proliferator-activated receptor , gamma (PPARγ)是細胞核內類固醇受體之一，PPARγ被認為與脂質代謝有關。綜合上述而知，ApoE、5-HT2C ､PPARγ可能與代謝症候群有些關連性存在。因此本研究主要探討精神分裂症代謝症候群與ApoE、5-HT2C ､PPARγ之基因多型性是否有關連性存在。
研究方法：本研究之研究對象為台灣某醫院之精神分裂症患者，收集300位經精神科專科醫師依據DSM-IV診斷為精神分裂症或情感性精神分裂症患者之年度健檢資料共350位，再依據2005年「美國國家膽固醇教育計劃成人治療指引第三版之修訂版（updated NCEP-ATP III）」的代謝症候群診斷標準（亞洲）由醫師作代謝症候群與否之診斷，於99年收案時共計得202位病患(其中61位為Mets及141位為non-Mets) 分析PPARγ Pro12Ala基因；100年收案2組，第一組共計得224位病患 (其中68位為Mets及156位為non-Mets) 分析ApoE基因，第二組共計得226位病患 (其中69位為Mets及157位為non-Mets) 分析HTR2C基因。利用PCR-RFLP的技術方法分析ApoE、5-HT2C ､PPARγ基因型分佈，再利用統計軟體進行統計分析。
研究結果：1.精神分裂症患者之PPARγ Pro12Ala及HTR2C Cys23Ser基因多型性不論是P allele或A allele與C allele或S allele的出現，皆不會導致代謝症候群的風險增加；2.精神分裂症患者之ApoE基因多型性在E3/E4型較E3基因型發生代謝症候群的風險增加2.431倍；3.ApoE基因多型性在E4單套體型較E3單套體型發生代謝症候群的風險增加 2.092倍，皆達統計學上顯著差異的標準(P<0.05)。
英文摘要
Background and Purpose: The schizophrenic patients have higher prevalence of metabolic syndrome than general population. Some association between HTR2C and metabolic syndrome was found. Apo E and Peroxisome proliferator-activated receptor are associated with the lipid metabolism. The aim of this study was to determine the association between gene polymorphisms and metabolic syndrome.
Methods: After obtaining the permission of IRB, we plan to enroll 300 schizophrenic or schizoaffective patients in Jianan Mental Hospital. The diagnosis of metabolic syndrome is done by updated version of NCEP-ATP III for Asia. The polymorphism is checked by PCR-RFLP. All these 300 patients receive genetic polymorphism analysis of the ApoE polymorphism、HTR2C polymorphism and PPARγ polymorphism.
Results: There is no association between PPARG Pro12Ala and HTR2C Cys23Ser with metabolic syndrome in enrolled 202 and 226 patients.ApoE polymorphism in our population 226 patients (69.12% for E3/E3 , 25% for E3/E4 , 2.94% for E2/E3 , 2.94% for E2/E2) were in agreement with those reported for Romania and other Caucosion populations. The distribution of ApoE alleles indicated a higher frequency of E4 (P<0.05) in groups E2 and E3.
Conclusion: Correlations of apoE genotypes with these markers indicate that the E4 allele is an independent risk factor for metabolic syndrome.
Keyword：Schizophrenia、Metabolic Syndrome、Gene Polymorphis
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