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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
Analysis of the Biological and Psychosocial Factors Related to Depression during Pregnancy
No full text誌 謝 iii
中文摘要 iv
Abstract vi
Contents viii
Tables and Figures xi
Appendixes xiii
Abbreviation xiv
Chapter 1 Introduction
1-1 Influence of prenatal depression on mothers and fetuses 1
1-2 Correlation of pregnant stress, social support, and prenatal depression 3
1-3 Biomarkers of depression 4
1-4 Correlation of BDNF and genotypes on psychiatric diseases 4
1-5 Correlation of S100B and genotypes on psychiatric diseases 6
Chapter 2 Materials and methods
2-1 Research Design 8
2-2 Subjects 8
2-3 Data collection 8
2-4 Measurement of questionnaires 9
2-4-1 Edinburgh Postnatal Depression Scale (EPDS) 9
2-4-2 Pregnancy Stress Rating Scale (PSRS) 9
2-4-3 Maternity Social Support Scale (MSSS) 10
2-5 Preparation of blood sample 10
2-5-1 Blood/serum withdrawal 10
2-5-2 Serum preparation 10
2-6 Analysis of genotypes 11
2-6-1 DNA isolation 11
2-6-2 Genotyping 12
2-6-3 Polymerase chain reaction (PCR) amplification 12
2-6-4 SNP analysis 13
2-7 Measurement of biomarkers 14
2-7-1 Serum BDNF 14
2-7-2 Serum S100B 15
2-8 Statistical analyses 17
Chapter 3 Results
3-1 Subject characteristics 19
3-2 Grouping 19
3-3 Correlation between extrinsic factors and prenatal depression 19
3-4 Measurements of the prenatal depression during pregnancy 20
3-5 Correlation between SNPs and prenatal depression 21
3-6 Allele frequencies of BDNF and S100B polymorphisms 22
3-7 Comparison of serum BDNF and S100B levels 23
3-8 Haplotype frequency estimation of S100B 24
Chapter 4 Discussion 25
Chapter 5 Conclusion 33
References 34
Tables and Figures 46
Appendixes 66
Biography of author 83[[abstract]]懷孕會造成孕婦身體心像的改變，也會加重孕婦的壓力，如果能夠早期發現，提供良好的心理及社會支持，可以降低孕婦因為懷孕造成的不適，也可以降低孕期憂鬱對孕婦及胎兒造成的影響。孕期憂鬱在歐美國家是熱門的研究主題，而國內針對孕期憂鬱方面的研究較少。本研究目的為探討單核苷酸多型性、特定生物指標變化、心理壓力及社會支持程度與孕期憂鬱的相關性。研究採便利取樣，以台南一家醫學中心與一家婦產科診所懷孕22~29週的孕婦為研究對象，自2010年5月至11月共收取45位個案。本研究的測量指標涵蓋心理社會以及生理兩部分。心理社會指標包含孕期憂鬱、心理壓力與社會支持，分別以愛丁堡產後憂鬱量表、孕期心理壓力量表以及孕婦社會支持量表測量，測量時間共計兩次，分別在第二孕期及第三孕期；生理指標包括BDNF與 S100B基因單一核甘酸多型性 (single nucleotide polymorphisms; SNPs) 和其血中濃度。於第二孕期同時抽血8 mL以量測生理指標。研究結果顯示在第二孕期，孕婦的年齡、工作狀況以及心理壓力與憂鬱呈顯著相關；而第三孕期的心理壓力、社會支持與憂鬱呈顯著相關。進一步分析資料，發現孕婦年齡較為年輕以及無工作者在懷孕期間較易出現憂鬱；至於社會支持方面，其在第二孕期與憂鬱未達顯著相關，而在第三孕期則與憂鬱達統計上顯著的負相關。生物指標的部分，本研究評估BDNF及S100B基因的單核苷酸多型性，並檢測血清BDNF與S100B的含量。在SNP部份，結果顯示孕期憂鬱與BDNF和S100B的SNP未達顯著相關。血清S100B的含量則與孕期憂鬱間呈正相關。總結本研究結果顯示，在第二孕期，孕婦的年齡、工作狀況、心理壓力以及血清中的S100B含量與孕期憂鬱間有顯著相關；而在第三孕期，心理壓力以及社會支持與孕期憂鬱間有顯著相關。期望本研究結果能夠當作一個懷孕期間憂鬱篩選的方法，進而減少孕期憂鬱對於孕婦及胎兒的不良影響。
It has been known that pregnancy causes changes in body image and increasing stress on women. If the signs of prenatal depression can be detected at earlier phase of pregnant period, health care professionals can provide the information to help relieve the discomfort and decrease the influence of depression on pregnant women and fetus. Prenatal depression is a popular research topic in the United States and Europe, but not in Taiwan. The purpose of this study was to investigate the correlation of prenatal depression with SNPs, biomarkers, psychological stress, social support. In this study, we collected samples from 45 subjects who were 22 to 29 weeks of pregnancy from May to November, 2010 in a medical center and an obstetrics and gynecology hospital in Tainan. In this study, we evaluated two portions including the psychosocial factors (prenatal depression, psychological stress and social support) and physiological factors (single nucleotide polymorphisms, SNPs, and biomarkers). We used three scales to measure the psychosocial factors including Edinburgh Postnatal Depression Scale (EPDS), Pregnancy Stress Rating Scale (PSRS), and Maternity Social Support Scale (MSSS) which were measured twice in 2nd and 3rd trimester during pregnancy; meanwhile, we withdrew 8 mL of blood to measure physiological factors in 2nd trimester during pregnancy. The results showed that age of pregnant women, work status, and psychological stress significantly correlates with depression during 2nd trimester of pregnancy. Psychological stress and social support significantly correlate with depression during 3rd trimester of pregnancy. Then, the results further revealed that younger age and unemployed status of women positive-significantly correlates with depression during pregnancy. However, social support negatively correlates with depression during 3rd trimester of pregnancy but not significantly in 2nd trimester. In biological factors, we evaluated the SNPs of brain-derived neurotrophic factor (BDNF) and S100B genes and the serum levels of those two proteins. These results of this study also revealed that no significant correlation between SNPs of S100B and BDNF and depression during pregnancy. However, we discovered positive correlation between serum S100B and prenatal depression. Taken together, these results revealed that age of pregnant women, work status, psychological stress, and serum levels of S100B significantly correlates with depression during 2nd trimester. Furthermore, this study also pointed out that the psychological stress and social support significantly correlates with prenatal depression during 3rd trimester. In conclusion, these results suggested that this study provides a screening method for depression patients during pregnancy to prevent the adverse effects of prenatal depression on mothers and fetuses, especially, the cases in Taiwan
In Vitro Study of Tamoxifen–induced growth inhibition in estrogen–receptor negative Glioma cells
No full text目錄
摘要.....................................................ⅰ
英文摘要..................................................ⅱ
誌謝.....................................................ⅲ
目錄.....................................................ⅳ
圖表目錄..................................................ⅴ
第一章 前言
第一節 腦瘤...........................................1
第二節 細胞週期........................................4
第三節 細胞凋亡........................................5
第二章 材料方法
第一節 細胞解凍.......................................8
第二節 細胞培養.......................................8
第三節 繼代培養.......................................9
第四節 細胞存活率分析..................................9
第五節 反轉錄聚合酵素連鎖反應 ..........................11
(reverse transcription polymerase chain reaction:RT-PCR)
第六節 西方墨點法(Western Blotting) .................13
第七節 基質金屬蛋白酵素分析(Gelatin Zymography) .......15
第三章 實驗結果
第一節 腦瘤細胞經由RT-PCR偵測Estrogen receptor mRNA表現
............................................19
第二節 Tamoxifen在不同藥物濃度處理24~48小時後腦瘤細胞存活率的變化(10%FBS與0.5%FBS)。
............................................20
第三節 Tamoxifen在不同藥物濃度處理24~48小時後達到50%細胞致死率的濃度分析(IC50)
............................................20
第四節 Tamoxifen在相同藥物濃度處理6~24小時後腦瘤細胞型態變化
............................................20
第五節 腦瘤細胞經由Tamoxifen藥物處理後細胞增生蛋白質的變化
............................................21
第六節 腦腫瘤細胞經由Tamoxifen藥物處理後細胞轉移蛋白的變化
............................................21
第四章 結論與討論....................................22
參考文獻...............................................28
圖表..................................................37
圖表目錄
表一 細胞週期........................................4
表二 細胞週期抑制蛋白..................................4
表三 引子合成........................................12
表四 蛋白質電泳膠片製作................................16
Fig.1 以RT-PCR技術分析腦瘤細胞中雌激素受體的表現。
............................................37
Fig.2 Tamoxifen對腦瘤細胞體外生長抑制的影響。
............................................38
Fig.3 Tamoxifen 達到50% 細胞致死率的濃度分析(IC50)。
............................................41
Fig.4 腦瘤細胞經Tamoxifen藥物處理後於不同時間點細胞型態的變化。
............................................44
Fig.5 腦瘤細胞經由Tamoxifen藥物處理後細胞增生蛋白質的變化。
............................................45
Fig.6 腦瘤細胞經由Tamoxifen藥物處理後細胞轉移蛋白質的活性變化。
............................................46[[abstract]]中文摘要
根據行政院衛生署統計，近年來惡性腫瘤為十大死亡原因之首。
本篇研究所探討的腫瘤屬於高度惡性腫瘤:神經膠原母細胞瘤(Glioma)。
雖然成人惡性腦瘤占惡性腫瘤2~3%，比例不高，但通常有三分之二的病人存活率不會超過一年，加上大腦是人體指揮中樞，所以腦瘤在腦部所引起的臨床症狀，會對患者造成嚴重的後果，而成為臨床醫療與病人照顧的重要課題。
生理結構中正常腦組織帶有一層薄膜，這層薄膜被稱為血腦障壁。
此障壁可阻止有害物質進入腦中，加上腦瘤抗原的免疫原性弱，不易引起強烈的免疫反應，又由於血腦障壁的存在，抗癌免疫反應不易落實至腦內，惡性腦瘤的藥物治療選擇因此有限。
由於血腦障壁的特性，化學類藥物只能靠藥物的脂溶性通過血管內膜細胞再進入腫瘤細胞產生作用，因此選用類荷爾蒙類的藥物Tamoxifen來做為實驗用藥。
臨床上Tamoxifen用來阻斷雌激素受體作用，而抑制乳癌細胞生長，並可用來做為乳癌的化學預防治劑。
而Tamoxifen藥物結構與雌激素相似，本實驗主要是研究Tamoxifen是否可以有效抑制腦腫瘤細胞生長，包含其作用機轉的初步分析。期盼提供臨床上腦腫瘤患者不同的藥物合併治療與化學預防的選擇方向。
根據我們的實驗結果發現Tamoxifen的確會造成腦腫瘤細胞GBM8901與DBTRG的死亡，並且是藉由不表現雌激素受體的路徑所造成，這種死亡方式可能是藉由細胞凋亡的機制達成。
此外，在腦腫瘤細胞株中，經Tamoxifen處理後，與腫瘤增生有關的增生蛋白Rb、PCNA、Akt2也都有顯著的下降，並且參與腫瘤轉移的轉移蛋白MMP-2活性亦有明顯的抑制。
因此，在體外研究當中Tamoxifen確實具有抑制腦腫瘤細胞生長的能力。
綜合以上實驗結果，Tamoxifen是一種具有潛力應用在腦瘤患者化學預防與治療用藥的選擇。
Abstract
In recent years, malignant tumor is the leading cause of death in Taiwan. We concentrate our attention on the malignant tumor: Glioma. Although Glioma is not highly occurred in the malignant tumors for only 2~3%, one year survival percentages are less than 2/3. Besides, the brain controlling our daily activities, therefore Glioma will cause a serious consequence to the patients. How to treat Glioma patients with complementary medicine to increase life quality becomes an important topic in clinical medical service.
The physiological structure of normal brain has a thin film called the Brain Blood Barrier. This barrier can make brain in an aseptic environment and avoid any microorganism infection. In addition, the immune activity of Glioma is too weak to cause intense immune response and the anticancer treatment is not easy to carry out to the brain because of Brain Blood Barrier. These reasons make pharmacological treatment of Glioma is therefore limited.
Clinical medicine chose in brain tumor treatment can only depend on the characteristic of fat soluble in order to get through Brain Blood Barrier. Thus we used Tamoxifen, originally synthesized as an anti-estrogen to inhibit proliferation of estrogen receptor-positive breast cancer cells. It can also inhibit proliferation in estrogen receptor-negative cancer cells. However, the molecular basis of such estrogen -independent growth inhibition is still unknown. In this study, we used estrogen receptor-negative Glioma cells to clarify the inhibitory effect of Tamoxifen in vitro.
According to our studies, we found that Tamoxifen can induce Glioma cell death in estrogen receptor-independent manner. This maybe through apoptosis pathway and needed more advantage analysis. After Tamoxifen treatment, we also found that proliferation-associated proteins including Rb、PCNA、Akt2 were decreased. The enzyme activity of invasion protein MMP-2 was also significant inhibited by Tamoxifen. These results demonstrate that Tamoxifen can inhibit estrogen receptor-negative Glioma cell growth in vitro by decreasing proliferation proteins expression and inhibiting MMP-2 activity and suggest that Tamoxifen has the potential in chemo-prevention and treatment of Glioma patients
