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素食飲食與血中同半胱胺酸、C-反應蛋白、脈衝波速之相關性探討
No full text[[abstract]]本研究以慈濟委員及慈誠聯誼會的20歲以上會員為研究對象，素食592人、偶而素食812人及非素食678人，共計2082人，檢測脈衝波速並蒐集問卷、血液生化、疾病史與體位測量等資料。以脈衝波速大於1400 cm/sec定義為「高脈衝波速」，探討素食飲食型態對慢性疾病與脈衝波速的影響
台灣地區外籍、本國籍、原住民婦女之子宮頸抹片篩檢行為
No full text[[abstract]]本研究之目的為了解台灣婦女參與子宮頸抹片篩檢之現況及運用醫療服務利用行為模式（Andersen's health-service utilization model）探討影響婦女參與子宮頸抹片篩檢之相關因素
科學工業園區臭味污染評估研究
No full text[[abstract]]本研究目的為評估高臭味污染工業區臭味污染之發生頻率、強度及濃度，並檢測環境因子及相關空氣氣狀與粒狀污染物，以評估科學工業區臭味污染情形
消防人員因職業暴露影響肺功能及腸胃道症狀之評估研究
No full text[[abstract]]根據內政部消防署統計資料顯示，台灣地區火災的主要肇因包括：電器設備、菸蒂、人為縱火、機械設備、爐火烹調、敬神掃墓祭祖等。火災現場空氣中有害物質係以粉塵(dusts)、燻煙(fumes)、霧滴(mists)等粒狀物以及氣體或蒸氣之狀態存在，而消防人員因長期出勤救火可能暴露在高濃度的有害物質中，進而影響身體健康，導致肺功能的損失或是可能產生肺癌及其他腸胃道疾病
Develop urea biosensor based on an ammonium electrode
Get PDF口試委員會審定書
授權書 i
目錄 ii
圖表目錄 iv
中文摘要 v
英文摘要 vii
誌謝 x
第一章緒論 -1-
1-1 研究背景 -1-
1-2 研究目的與動機 -3-
1-3 論文架構 -4-
第二章 理論分析與探討 -6-
2-1生物感測器之種類功能介紹 -6-
2-2酵素固定化技術 -8-
2-3載體 -9-
2-3-1 氨與銨之簡介 -10-
2-3-2 銨離子選擇電極之選擇 -11-
2-3-3 銨離子選擇電極之特性參數探討 -11-
2-3-4 銨離子選擇電極離子干擾分析 -14-
2-4延伸式閘極感測場效電晶體之原理 -15-
2-5 尿素含量在臨床之探討 -16-
2-6 文獻探討 -17-
第三章 實驗方法與材料 -18-
3-1材料 -18-
3-2量測系統 -20-
3-3儀器與器材 -21-
3-4 感測器的製備 -22-
3-4-1 可繞性感測器的製備 -22-
3-4-2 銨離子感測電極的製備 -23-
3-4-3 銨離子尿素感測器的製備 -24-
第四章 結果與討論 -25-
4-1銨離子感測膜反應校正曲線和標準差 -26-
4-2銨離子電極對不同濃度尿素的反應 -28-
4-3不同緩衝液對於不同濃度尿素的反應趨勢 -29-
4-4 Tris是含EDTA對於尿素反應的比較 -31-
4-5 Tris有無含NH4Cl對於尿素反應的影響 -33-
4-6尿素濃度區間反應之選擇性 -35-
4-7銨離子選擇電極於尿素濃度區間4~8mg/dl之反應特性 -36-
4-8 銨離子選擇電極於尿素濃度區間1-2mg/dl之反應特性 -37-
第五章 結論 -39-
參考文獻 -41-[[abstract]]氨（NH3）是一種大氣或自然界水體中無所不在的化合物，是所有含氮物種中化學結構最簡單的化合物。在pH值小於8 的情況下，氨溶於水之後會和水分子作用而大部分以銨離子（NH4+）的形式存在，若是氨的含量過多之時，將會讓水體優養化，進而導致水體生態失去平衡。
在人體內，血漿中正常之尿素濃度為18-36 mg/dl，而臨床上尿素測量常以其含氮量表示，稱為尿素氮，1 mg/dl尿素氮相當於2.14 mg/dl尿素濃度，尿素氮會因其人體狀態之不同而顯現尿素濃度之不同，也因此量測血液中之尿素含量在臨床上是具有其意義的。
為檢測其環境(氨態氮)以及人體(尿素氮)之銨離子需求，本論文研究之電極所達到之最低檢測限如完善開發，將可對更低濃度尿素之環境或者人體血液尿液進行更準確之量測。
本論文利用延伸式ITO/PET離子感測場效電晶體製作耐用型的銨離子選擇電極，探討各個參數對於整體反應之影響，經過一些實驗之結果可得知銨根離子可偵測濃度為10-5 ~ 1M 之範圍，線性範圍約為0.99，平均感測度為55.09 mV / pNH4+ ，緩衝液部分利用PB以及Tris進行量測，利用Tris緩衝液可以使量測結果趨於穩定，在收集文獻過程中有些許文獻指出在緩衝液內加入EDTA會有更好的感測度，因此進行了加入及不加入EDTA的實驗，但發現不加入EDTA之反應更加的靈敏，因而採用不加入EDTA繼續進行研究，為了顯現此感測器的價值，也就是感測限比傳統型尿素感測器低，因而進行尿素濃度區間展開之實驗(1mg/dl~8mg/dl)，在此展開結果中可得知此電極在尿素濃度在低下的狀況下所得到之反應會比一般感測器來的更加靈敏，檢測限更低，在人體方面，未來可利用稀釋此種方式來對正常人體之尿素或血液將其稀釋至最佳檢測濃度，將可得到最為靈敏且可靠之反應結果。
在環境檢測中，莫過於水質汙染最為嚴重，因此對於水體的監控更能看出此感測器的價值，因水體中存在之銨離子所產生之尿素是微乎其微的，因此感測器檢測限低，可針對銨離子產生之低濃度尿素進行監測，獲得更加靈敏且可靠的結果，因此可用此感測器來對自然水體環境進行監控。
此感測器有著比一般感測器更低之檢測限度以及更佳的靈敏度，對於未來要監測環境中低濃度的尿素可以更迅速且靈敏的檢出，也可針對人體之血、尿液來進行監測，此電極未來如完善的開發，將可對環境及人體有著一大貢獻。
關鍵字:銨，生物感測器，銨離子選擇電極
論文外文摘要： Ammonia (NH3) is a compound that exists ubiquitously in the atmosphere and natural water and is the compound having the simplest chemical structure among the nitrogen-containing species. In the case of pH value below 8, as ammonia dissolves in water, it reacts with water molecules to form ammonium ions (NH4+) mostly. When there is too much ammonia dissolved in water, the water becomes eutrophic, leading to the aquatic ecological imbalance.
In the human body, the normal urea concentration in the plasma is 18-36 mg/dl whereas in the clinical practice urea is most often measured in term of the amount of nitrogen, namely the urea nitrogen. The concentration of urea nitrogen of 1 mg/dl is equivalent to a concentration of urea of 2.14 mg/dl. Urea nitrogen varies according to the human body condition and reflects on the variation of urea concentration. Therefore, measuring the concentration of urea in blood can be a meaningful method to monitor the body condition in the clinical practice.
For the demand of detecting the ammonium ions of the ammoniacal nitrogen in the environment and of the urea nitrogen in the human body, if the lowest detection limit of the electrode described in this research publication can be fully developed, further lower concentrations of urea in the environment or human blood and urine can be measured more accurately.
In this publication, the extended ITO/PET ion-sensitive field-effect transistors are used to produce the durable ammonium ion-selective electrodes for investigating the impact to the overall reaction affected by each parameter. According to the results of several experiments, the concentration of ammonium ion in the range between 10-5 ~ 1M is detectable with a linear range of about 0.99 and the average sensibility is 55.09 mV / pNH4+. As for the buffer solution, PB and Tris buffer solutions are used for measurement whereas the use of Tris buffer can stabilize the measuring results. While researching the references, some references indicate that the sensitivity can be better by adding EDTA in the buffer solution. Therefore, experiments with EDTA added and without EDTA added in the buffer solutions were both conducted. However, the results show that the experiments without adding EDTA have better sensitivities to the reactions. Since then, the experiments proceeded without EDTA. In order to demonstrate the value of this sensor whose lowest detection limit is lower than that of a traditional urea sensor, experiments thus have been conducted with the interval expansion of urea concentration (1mg/dl ~ 8mg/dl).
The expansion result shows that this electrode is more sensitive than a regular sensor in reacting to lower urea concentrations, and the detection limit of this electrode is also lower. For monitoring the human body condition, to apply this dilution method to dilute the urea or blood from a normal body to the most suitable concentration level for measurement will bring the most sensitive and reliable reaction result in the future.
Nothing is more serious than the water pollution in the detection of the environmental hazards. Therefore, this sensor is even more valuable when it comes to the monitoring of the water bodies. Since the urea produced by the ammonium ions in the water bodies is found in trace quantities, the lower the detection limit of the sensor is, the more sensitive and reliable result the sensor can produce while monitoring the extreme low concentration of urea produced by the ammonium ions. Therefore, this sensor can be used to monitor natural water bodies and environments.
This sensor has a detection limit lower than that of a regular sensor and is of better sensitivity. This sensor can more quickly detect the low concentrations of urea exists in the environment in the future and can monitor the blood and urine in human bodies. If this electrode can be fully developed in the future, it can bring significant contribution to the environmental protection and human body wellness.
Keywords: Ammonia, Ammonium biosensor, Ammonium ion-selective electrod
Benzene exposure and urinary metabolite S-PMA in chemical industry workers in Taiwan
Get PDF致謝 I
中文摘要 II
Abstract III
目錄 IV
圖目錄 VI
表目錄 VII
第一章 前言 1
第一節 研究背景與動機 1
第二節 研究目的 2
第二章 文獻探討 3
第一節 苯之特性及來源 3
第二節 苯之健康效應 4
第三節 苯之人體代謝途徑 5
第四節 苯之生物偵測 6
第五節 苯作業環境暴露容許濃度標準 8
第六節 苯暴露的環境及職業衛生相關研究 9
第三章 材料與方法 13
第一節 實驗材料與設備 13
第二節 研究架構 15
第三節 作業環境之採樣方法 16
第四節 生物偵測之採樣方法 20
第五節 統計分析 27
第四章 結果 28
第一節 化學廠苯作業環境測定結果 28
第二節 化學廠受測勞工問卷調查及尿中S-PMA生物偵測結果 28
第三節 苯作業環境與生物偵測之相關性 36
第五章 討論 37
第六章 結論 42
參考文獻 44
附錄 62
附錄一、問卷 62[[abstract]]本研究目的為探討台灣化學業勞工職業苯暴露與尿中苯主要代謝物苯基硫醇酸(S-phenylmercapturic acid, S-PMA) 濃度及相關性情形。苯作業環境暴露評估是選擇3家有使用苯之化學公司，每公司1~2個廠區，每廠區選擇3~4處採樣點，進行連續3個工作天的作業環境空氣採樣，總共採集樣本數為45個；生物偵測方面，以作業區內苯暴露勞工於上班前與下班後各收集1次尿液樣本，檢測尿中S-PMA濃度情形，總計受測勞工49人，尿液樣本數為98個；並針對勞工進行問卷調查，項目包括基本資料與工作概況、生活與飲食習慣及防護具使用情形等。研究結果顯示，在環境測定方面，有8.89 %樣本可測得量化之苯濃度，其濃度範圍為0.667~1.448 ppm，其餘樣本濃度皆低於本研究方法偵測極限0.423 ppm。受測勞工上班前尿中S-PMA濃度為0.175±1.55 μg/g cr.，下班後尿中S-PMA濃度為0.373±1.86 μg/g cr.，下班後尿中S-PMA濃度有顯著增加 (p<0.05)，但皆未超過美國工業衛生師協會 (American Conference of Governmental Industrial Hygienists, ACGIH) 的生物暴露指標 (Biological Exposure Indices, BEI) 建議值25 μg/g cr.；此外，上班前尿中S-PMA濃度會顯著受到受測者吸菸行為及二手菸暴露情形影響，而下班後尿中S-PMA濃度則未受吸菸行為及二手菸暴露因素所干擾，顯示下班後尿中S-PMA濃度以職場暴露為主要來源。本研究發現作業環境苯濃度較高的作業區，其作業勞工尿中S-PMA濃度亦有較高的趨勢，顯示受測勞工職業苯暴露以作業環境空氣為主要來源。綜合以上所述，尿中S-PMA濃度對於評估化學業勞工職業性低濃度苯暴露是良好的生物偵測指標。
The purpose of this study was to explore the benzene exposure and urinary metabolites S-phenylmercapturic acid (S-PMA) in chemical industry workers. Benzene exposure was executed in three chemical companies, in which 3-4 sites in each working area were selected, and the total number of air sample was 45. Urine samples were collected from 49 workers before and after work, and total number of urine sample was 98. Questionnaire, including demography, work history, lifestyle, habits and personal protective equipment usage, was also evaluated for the workers. The results showed that benzene could be quantified in 8.89 % of air samples, and the concentration were 0.667~1.448 ppm; the other samples were below the detection limit of 0.423 ppm. The urinary concentration of S-PMA were 0.175±1.55 before work and significantly increased to 0.373±1.86 after work (P<0.05); all the urinary concentrations of S-PMA did not exceed the biological exposure indices (BEI) of 25 μg/g cr. proposed by American conference of governmental industrial hygienists (ACGIH).This study also found the smoke habit and environmental tobacco smoke exposure of the workers could affect the pre-shift urinary S-PMA concentration, but did not affect the post-shift urinary S-PMA concentration. Meanwhile, the workers exposed higher concentrations of benzene in work area could be detected higher concentrations of urinary S-PMA, and this indicated the inhalation was the main benzene exposure route for the workers. In conclusion, urinary S-PMA was a good indicator to evaluate the occupational exposure for low concentration of benzene in work fields in chemical industries
Study of Functions of Different C-terminal Domains of Integrin-Associated Protein
No full textContents i
Figure contents iv
致謝 v
Abbreviation vi
中文摘要 1
Abstract 3
Chapter 1 Introduction
1.1 Integrin associated protein/CD47 5
1.2 Expression of CD47 in tissues 6
1.3 PLIC is the ligand of CD47 7
1.4 Functions of CD47 7
1.4.1 Roles of CD47 in neurite outgrowth 8
1.4.2 Roles of CD47 in phagocytosis 9
1.5 Specific aims 9
Chapter 2 Materials and Methods
2.1 Cell Culture 11
2.2 Cloning of different isoforms of CD47 11
2.2.1 RT-PCR 11
2.2.2 Cloning of different isoforms of CD47 into TA vector 12
2.2.3 Transformation 12
2.2.4 Plasmid DNA extraction 12
2.2.5 DNA sequencing 13
2.2.6 Preparing cDNA of CD47 isoforms 14
2.2.7 Construction of different full-length cDNA of CD47
isoforms into pCMSEGFP vector 14
2.3 Confirmation of different isoforms of CD47 15
2.4 Transfection of cDNA to IMR32 cells 15
2.5 Measurement of neurite outgrowth 16
2.6 Transfection of cDNA to RAW 264.7 cells 16
2.7 Bead opsonization 17
2.8 Phagocytosis 17
2.9 Statistical analysis 17
Chapter 3 Results
3.1 Identification of new alternative splicing isoforms of
CD47 mRNAs 19
3.2 Confirmation of alternative splicing isoforms of CD47
mRNAs using isoforom-specific primers 19
3.3 Gene structures of CD47 isoforms 20
3.4 Predicted translated amino acid sequences of CD47
isoforms 20
3.5 Isoforms of CD47 play roles in neurite outgrowth 21
3.6 Isoforms of CD47 do not involve in phagocytosis 21
Chapter 4 Discussion
4.1 Differences in CD47 isoforms 23
4.2 Role of CD47 in neuronal differentaion 24
4.3 Role of CD47 in phagocytosis 24
4.4 Conclusion 25
Chapter 5 References 26
Figure 33
作者簡歷 45
Figure contents
Fig. 1 Identification of isoforms of CD47 mRNAs using common
primers 33
Fig. 2 Alignment of alternative splicing regions of CD47 cDNAs 34
Fig 3 Identification of isoforms of CD47 mRNAs using
isoform-specific primers 35
Fig 4 A scheme illustrating the predictive gene structures of different
CD47 isoforms. 36
Fig 5 Alignment of the predicted amino acid sequence of various CD47
isoforms 37
Fig 6 Overexpression of CD47 cDNAs increases neurite outgrowth in
IMR32 cells 38
Fig 7 Images of phagocytic cells with low level of phagocytic ability 39
Fig 8 Images of phagocytic cells with medium level of phagocytic
ability 40
Fig 9 Images of phagocytic cells with high level of phagocytic ability
41
Fig10 Statistics of phagocytic cells transfected with different constructs
42
Fig 11 Statistic diagram for percentage of phagocytic cells 43
Table 1 Primers used in this study 44[[abstract]]CD47 又稱為 integrin-associated protein (IAP)最早被發現是因與整合素 αvβ3 相聯結而命名。CD47 在細胞外有一個免疫球蛋白變異區類似區域(IgV-like domain)、五次穿膜片段以及有一個位在蛋白質C 端的多樣性剪輯區域。在人類、老鼠與豬中已經發現 CD47 mRNA有五個不同多樣性剪輯片段。這些剪輯形式在基因和胺基酸序列具有高度保留特性，而 CD47 不同 isoforms 的 mRNA 在不同組織中有不同的表現量。近年來，科學家發現在老鼠中 CD47 基因的表現是與嫌惡逃避記憶形成有關。無論如何，CD47 不同 isoforms 的表現和角色至今仍未清楚。直到現在 CD47 isoforms 有多少種仍然不知道。因此我們目的是要研究在神經纖維母細胞 IMR-32 中 CD47 是否還有其他不同的 isoforms 以及探討不同 isoforms 的功能。在本篇研究中，我們使用反轉錄反應與聚合酶連鎖反應(RT-PCR)的方法在神經纖維母細胞中發現有另外五種不同 isoforms。接著我們針對不同新發現 isoforms 區域設計專一性的引子，使用反轉錄反應產物進行聚合酶連鎖反應來進一步確認這些新發現的 isoforms。為確認這些isoforms 的功能，我們選殖完整的 cDNA 序列，轉殖在神經纖維母細胞株 IMR-32 與單核球細胞株 RAW 264.7 中，我們證實 CD47 isoform 2、4、8 與 10 會促進神經突生長，但不會影響吞噬作用。綜合以上的結果，我們發現五種不同 CD47 的 isoforms，不同 CD47蛋白 C 端的區域形式可能不會影響神經纖維母細胞神經突生長及單核吞噬細胞的吞噬作用。這些結果可幫助我們對 CD47 蛋白有更深入的了解，以釐清其在不同組織的角色。
CD47, also named integrin-associated protein (IAP), was originally identified in association with integrin αvβ3. CD47 contains an extracellular IgV-like domain, five membrane-spanning segments and an alternatively spliced C-terminus domain. It has been shown that five alternative splicing isoforms of CD47 mRNAs are expressed in cells of human, mouse and pig. These forms are highly conserved in cDNA and amino acid sequences and the expression levels of different isoforms of CD47 mRNAs are difference among tissues. In recent years, scientists discovered that the expression of CD47 gene is closely associated with inhibitory avoidance learning and memory formation in rats. However, the roles and expression of different isoforms of CD47 are still unclear. Hence we looked for other unknown splicing isoforms of CD47 in IMR-32 cells and studied their functions. In this study, we used reverse transcription-polymerase chain reaction and found five new isoforms of CD47 mRNA in IMR-32 cells. To further confirm these forms expressed in cells, we designed specific primers for new-found isoforms and used RT-PCR to reconfirm it. To investigate the functions of these forms, we cloned full-length cDNAs of different isoforms of CD47 and transfected them into neuroblastoma and macrophage cell lines, IMR-32 and RAW 264.7 cells, respectively. overexpression of CD47 isoform 2, 4, 8 and 10 markedly increased neurite outgrowth but did not affect phagocytic ability. Taken together, we found five new isoforms of CD47 mRNAs and named them isoform 6 to 10, respectively. We also found that C-termini of CD47 proteins may not affect neurite outgrowth in neuroblastoma cells and phagocytosis in monocytes. These findings help us further understand the functions of CD47 and their differential roles in different tissues