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Establishment of exposure assessment on human skin by solar simulated ultraviolet irradiance
No full text[[abstract]]工作職場與日常生活中，非游離輻射曝露之情形非常普遍，非游離輻射對生物體危害以紫外線為最，人體組織受紫外線曝露以皮膚及眼球為最主要的傷害器官。
本研究利用市售之簡易可攜式USB4000與Avantes紫外光譜分析儀，對UVB燈管光功率進行量測，模擬其太陽光照之紫外線輻射劑量，建立對人體皮膚細胞株之曝露評估研究。
針對紫外線曝露評估的描述及量測，尚未有準確及被認可的評估模式供為探討。紫外線曝露人體皮膚之輻射劑量以紫外線指數（UV Index）為參考指標，該參數根據「紅斑作用光譜曲線」之權衡，由「國際光照委員會（CIE）」公布之數值以呈現對人類皮膚的損害影響評估。本研究利用USB4000紫外光譜分析儀，量測紫外線B燈管光功率，並加權「紅斑作用光譜曲線」得知其UVI指數及模擬其太陽光照之紫外線輻射劑量，並運用 Avantes紫外線光譜分析儀，分析 UVA、 UVB燈管之穩定性及比較不同光譜分析儀之量測準確度。藉以判定穩定燈源之穩定性，作為人體皮膚細胞劑量曝露精準之依據。採用不同輻射劑量曝露於人類皮膚細胞株，觀察細胞誘發氧化壓力反應（ROS assay）、細胞毒性（LDH assay）、細胞存活率（MTT assay）之生化指標。由結果得知細胞接受 UVA刺激後3小時，進行流式細胞儀分析胞內螢光質(GMean)呈現下降，經過 UVB 刺激後3小時後，細胞胞內活性氧分子（ROS）呈現增加。在 UVB 60 mJ/cm2及300 mJ/cm2輻射劑量曝露下，可以明顯看出細胞傷害的變化。由於 UVB 持續的傷害導致細胞之活性氧分子及細胞存活率降低，細胞傷害也隨著時間增加而持續攀升。
本研究之紫外線暴露模式可提供光源光譜功率圖和光波長範圍，並對燈源之穩定性做進一步的確認，應用兩種不同光譜分析儀測試及佐證數據的正確性，可提供建立曝露燈源時應有更完整的量測模式，細胞暴露 UVB 的模式亦可提供完整的野外紫外線輻射曬傷及曝露效果評估。
UV radiation (UVR) exposure from the sun and artificial UV sources has been known to be dangerous to public health and it plays a major role in the formation of skin cancer and eye injury. The utilization of Radiometer to measure Ultraviolet radiation in skin exposure assessment is limited due to the lack of spectral irradiance and full wavelength scanning. In this study, two broadband spectrometer systems (USB 4000 and Avantes Miniature Fiber Optic Spectrometers) both containing cosine corrective detectors with spectral analysis software were utilized for erythemal UVR measurement and exposure does evaluation.
Various UVR doses from UVA and UVB lamps were applied to induce oxidative stress in human keratinocyte (HaCaT) cell line. The exprosure cultured cells showed that the cellular ROS (reactive oxygen species) and LDH (lactate dehydrogenase) increase, and in coincidence with cellular survival (MTT test) decrease upon UVB 60 mJ/cm2 exposure. The flow cytometric results demonstrated that the cellular anti-oxidative effects were induced at 3 hours after exposure to UVB 60 mJ/cm2 and higher dose of UVB 300 mJ/cm2.
The UVR exposure model provides the information of spectrophotographies and full wavelength range scanning. The UVR exposure model from UV lamps was further to apply to the environmental solar-simulation UVR exposure assessment from laboratories
Preparation and Antioxidative Activities of Germinated Brown Rice Rich with GABA
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目錄------------------------------------------------------------------------------i~xi
圖目錄-------------------------------------------------------------------------vii~ix
表目錄--------------------------------------------------------------------------x~xi
致謝--------------------------------------------------------------------------------xii
中文摘要-------------------------------------------------------------------xiii~xiv
英文摘要--------------------------------------------------------------------xv~xvi
縮寫表(Abbreviations)------------------------------------------------xvii~xviii
第一章、 緒論-----------------------------------------------------------------1~25
第一節、 前言--------------------------------------------------------------1~2
第二節、 文獻整理-------------------------------------------------------3~25
一、稻米的種類------------------------------------------------------------3
二、稻米之構造---------------------------------------------------------4~5
三、稻米之化學成份及營養功效------------------------------------6~7
四、稻米未來展望------------------------------------------------------7~8
五、發芽米--------------------------------------------------------------8~14
1.『發芽米』之定義-------------------------------------------------8~9
2. 發芽米之營養價值-------------------------------------------9~12
3. GABA的生成路徑-------------------------------------------12~13
4. GABA之功效------------------------------------------------------14
六、電解水------------------------------------------------------------14~16
七、甲殼素(Chitosan) ----------------------------------------------16~18
八、L-麩胺酸(L-glutamic acid) ----------------------------------18~19
九、香椿（Toona sinensis Roemor）------------------------------19~21
十、微生物對食品的影響------------------------------------------21~23
1.食物感染菌----------------------------------------------------21~23
(1)金黃色葡萄球菌（Staphylococcal）--------------------21~22
(2)仙人掌桿菌（Bacillus cereus）------------------------------22
2.食品安全指標菌----------------------------------------------22~23
(1)大腸桿菌（Escherichia coli）---------------------------22~23
(2)大腸桿菌群（Coliforms）-------------------------------------23
十一、自由基與人類疾病的關係---------------------------------24~25
1. 何謂自由基（Free radical）-------------------------------------24
2. 抗氧化物之作用---------------------------------------------24~25
第三節、研究目的與實驗設計----------------------------------------26~27
一、研究目的--------------------------------------------------------------26
二、實驗設計--------------------------------------------------------------27
第二章、材料和方法--------------------------------------------------------28~62
第一節、發芽米之製備-----------------------------------------------------28
一、 材料及儀器-------------------------------------------------------28
二、 方法------------------------------------------------------------29~30
第二節、 不同品種糙米之GABA含量分析¬---------------------30~32
一、 材料和儀器---------------------------------------------------30~31
二、 GABA之萃取與衍生化方法------------------------------31~32
第三節、發芽率試驗--------------------------------------------------------32
第四節、貯藏期試驗----------------------------------------------------32~57
一、材料---------------------------------------------------------------32~34
二、方法---------------------------------------------------------------34~57
1.稀釋液配製---------------------------------------------------------34
2.生菌數檢測----------------------------------------------------34~35
3.厭氧總生菌數檢測-------------------------------------------35~36
4.金黃色葡萄球菌數檢測-------------------------------------36~41
5.大腸桿菌數檢測----------------------------------------------41~48
6.大腸桿菌群檢測----------------------------------------------48~50
7.仙人掌桿菌數檢測-------------------------------------------50~57
8. pH值測定----------------------------------------------------------57
9.可滴定酸度測定¬¬---------------------------------------------------57
第五節、發芽米抗氧化之分析----------------------------------------58~61
一、 材料----------------------------------------------------------------58
二、 方法------------------------------------------------------------58~61
1. DPPH自由基清除能力測定-------------------------------58~59
2. Trolox當量抗氧化能力測定（Trolox equivalent antioxi- dative capacity,TEAC）-------------------------------------------59
3. 羥自由基（Hydroxyl Radical,．OH）分析--------------59~60
4. 總酚含量分析------------------------------------------------60~61
第六節、感官品評-----------------------------------------------------------62
第七節、統計分析-----------------------------------------------------------62
第三章、 結果與討論-----------------------------------------------------63~106
第一節、不同品種糙米之γ-氨基丁酸(γ-aminobutyric acid; GABA)含量分析----------------------------------------------------------63~65
一、發芽前糙米GABA含量分析--------------------------------------63
二、不同品種發芽後GABA含量分析---------------------------63~65
1.南稉育(TJB)-------------------------------------------------63~64
2.台稉糯3號(TJG3) ----------------------------------------------64
3.台南11號(TJ11) --------------------------------------------64~65
第二節、添加麩氨酸(L-Glutamic acid, G)和甲殼素(Chitosan, C)以及不同百分比香椿嫩葉粉(Toona sinensis Roemor; TS)進行發芽之南稉育(TJB)發芽米分析--------------------------66~68
一、GABA含量分析-----------------------------------------------------66
1. GC組之GABA含量分析----------------------------------66~67
2. GCTS(0.1 ％)、GCTS(1 ％)及GCTS(5 ％)之GABA分析----------------------------------------------67~68
二、發芽率試驗-----------------------------------------------------------68
第三節、南稉育(TJB)發芽米之貯藏期試驗------------------------68~73
一、總生菌（Total aerobic plate counts）-----------------------------69
二、厭氧總生菌（Total anaerobic plate counts）---------------70~71
三、大腸桿菌（Escherichia coli）---------------------------------------71
四、大腸桿菌群（Coliform Bacteria）-----------------------------71~72
五、金黃色葡萄球菌（Staphylococcus aureus）和仙人掌桿菌（Bacillus cereus）-------------------------------------------------72
六、pH值測定--------------------------------------------------------72~73
七、可滴定酸度測定-----------------------------------------------------73
第四節、南稉育(TJB)發芽米萃取物之抗氧化分析---------------74~77
一、DPPH之清除能力---------------------------------------------74~75
二、Trolox equivalent antioxidant capacity, TEAC測定----75~76
三、羥自由基分析---------------------------------------------------76~77
四、總酚含量分析--------------------------------------------------------77
第五節、感官品評-------------------------------------------------------77~79
第四章、結論--------------------------------------------------------------108~109
參考文獻------------------------------------------------------------------110~120
附錄一---------------------------------------------------------------------------121
附錄二---------------------------------------------------------------------122~125
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Young, D. H., H. Kohle, and H. Kauss. (1982). Effect of chitosan on membrane permeability of suspension cultured glycine max and phaseolus vulgaris cells. Journal of Plant Physiol., 70, 1449-1454.[[abstract]]本研究以台灣良質米台稉糯3號（TJG3）、台南稉11號（TJ11）及南稉育（TJB）作為製備發芽糙米之材料，使用之浸泡水分別為自來水(pH 6.8)及pH 5.5、pH 9.0之電解水三組，浸泡水溫為28 ℃、32 ℃及36 ℃，並分別浸泡4 hr、6 hr、8 hr及24 hr，其中4、6、8 hr之浸泡組，再以真空包裝，維持相同溫度至24 hr，進行γ-氨基丁酸(γ-aminobutyric acid, GABA)含量之分析。結果顯示南稉育pH 9.0 /36 ℃/6 hr組GABA含量最高，為17.79 mg/100 g，次佳組為pH 5.5 /28 ℃/6 hr，GABA含量為16.53 mg/100 g。另為了增加GABA產生量，添加L-麩胺酸(L-Glutamic acid)及甲殼素(Chitosan)之後，自來水組確實有顯著增加其GABA含量，由9.57 mg/100 g 增加至16 mg/100 g；而pH 5.5組較無顯著差異，加入L-Glutamic acid及Chitosan組並無太大變化。香椿劑量增加至1 %以上時，降低其GABA含量至3.1 ~5.69 mg/100 g，其發芽率由70 %降至11.33 %，表示香椿可能會抑制其糙米發芽，進而降低GABA之合成。
在貯藏期試驗方面得知，總生菌及厭氧總生菌之含量，在貯藏期21天中，菌數均由106 cfu/g增加至109 cfu/g左右，而其中主要是以含L-Glutamic acid, Chitosan, 及1 %香椿組的(GCTS(1 %))組菌數含量較多。在貯藏期中，菌數均由107 cfu/g增加至109 cfu/g，產生較多的乳酸菌而使菌數增加。在貯藏期第7天之後，大腸桿菌有慢慢增加之趨勢；大腸桿菌群則隨著貯藏期增加，菌數由102 MPN/g增加至107 MPN/g左右，主要是以GCTS(1 %)和GCTS(5 %)組之菌數產生最多。各組之pH值與可滴定酸呈負相關性。
在抗氧化方面，DPPH，羥自由基之清除能力及TEAC測定，都是以添加香椿劑量＞1 %之組別，具有較佳之抗氧化能力，這顯示其抗氧化能力係來自香椿。GCTS(5 %)組提升DPPH清除能力達40 %；TEAC測定方面，GCTS(1 %)及GCTS(5 %)組均具有0.6 mM以上之Trolox當量，幾乎是未添加香椿之2倍以上。羥自由基分析中添加香椿之組別同樣可降低其抑制50％所需之樣品劑量(IC50)，因此，GCTS (1 %)及GCTS(5 %)添加組中可能具有較多之抗氧化成份。在總酚含量分析中，則發現GCTS(5 %)組有顯著（P＜0.05）較高之總酚含量，為0.19 ±0.02 mg，由此可解釋其較強之抗氧化活性。
在發芽米炊煮成米飯並製作壽司進行感官品評試驗方面。結果三種樣品壽司米、發芽米及糙米中，以發芽米和壽司米之接受度較佳，糙米之口感較無法接受，所以其整體喜好顯著（P＜0.05）較低；但做成壽司後，可能其調味料及添加之食材影響其口感。所以發芽米和糙米之整體喜好度無顯著差異，但仍有發芽米較佳之趨勢。
This research use Taiwan Japonica Glutinous 3 (TJG3), Tainan Japonica 11 (TJ11) and Tainan Japonica Breed (TJB) brown rice, as the material to prepare germinated brown rice, will study the effect of three pH value water（tap water, pH 5.5, pH 9.0）, three water temperature（28, 32, 36 ℃）and different soak time 4, 6, 8, 24 hr, on γ-aminobutyric acid (GABA) formation. The result in the GABA content of TJB pH 9.0 / 36 ℃ / 6 hr is the highest (17.79 mg/100 g), the following is pH 5.5 / 28 ℃ / 6 hr (16.53 mg/100 g). Add L-Glutamic acid and Chitosan really sees a significantly increasing from 9.57 mg/100 g to 16 mg/100 g. PH 5.5 groups don’t show significantly difference. While increased the Toona sinensis Roemor dosage to above 1%, reduce GABA content from 3.1 mg/100 g to 5.69 mg/100 g, verify might Toona sinensis Roemor may inhibit brown rice germinated and then reduce the formation of GABA.
Analysis of Microorganisms, Total aerobic plate counts and Total anaerobic plate counts of all group except GCTS(1 %)(include L-Glutamic acid, Chitosan and 1 % Toona sinensis Roemor) increased from 106 cfu/g to 109 cfu/g over 21 days. Total aerobic plate counts and Total anaerobic plate counts of GCTS(1 %) increased from 107 cfu/g to 109 cfu/g. The MPN of E. coli was below
Development of oxidative stress damage model and protective effects of antioxidant agents in keratinocytes
No full text目錄
摘要 i
圖目錄 vi
表目錄 x
第一章 前言 - 1 -
1.1. 研究目的 - 2 -
第二章 文獻回顧 - 3 -
2.1. 紫外線輻射 - 3 -
2.2. 紫外線輻射分類 - 3 -
2.3. 紫外線對人體的影響 - 4 -
2.4. 活性含氧族群概述 - 7 -
2.5. 生物體內抗氧化系統 - 10 -
2.6. 紫外線暴露細胞模式 - 14 -
第三章、材料方法 - 17 -
3.1. 實驗儀器與藥品 - 17 -
3.2. 細胞培養 - 21 -
3.2.1. 細胞株 - 21 -
3.2.1.1. 老鼠皮膚細胞株 - 21 -
3.2.1.2. 人類角質層細胞株 - 21 -
3.2.2. 培養液的配置 - 21 -
3.3. 實驗設計 - 22 -
3.4. 皮膚細胞氧化壓力傷害模式建立方法試驗 - 24 -
3.4.1. UVB 燈管的劑量量測 - 24 -
3.4.2. 不同UVB 劑量對皮膚細胞傷害試驗 - 25 -
3.4.3. 皮膚細胞氧化壓力傷害模式建立之試驗 - 25 -
3.4.4. ROS assay - 26 -
3.4.5. Dihydroethidium assay - 28 -
3.4.6. 細胞毒性測試 - 29 -
3.4.7. 細胞存活率試驗 - 30 -
3.5. 探討抗氧化物對不同細胞株的保護能力模式建立方法實驗 - 32 -
3.5.1. 不同抗氧化物對皮膚細胞的毒性測試 - 32 -
3.5.2. 抗氧化物對不同細胞株的保護能力模式建立之試驗 - 32 -
3.5.3. 抗氧化壓力傷害能力評估指標 - 33 -
3.6. 統計分析方法 - 38 -
第四章 結果與討論 - 49 -
4.1. 皮膚細胞氧化壓力傷害模式建立方法實驗 - 49 -
4.1.1. UVB燈管的劑量量測 - 49 -
4.1.2. 皮膚細胞氧化壓力傷害模式建立之試驗 - 49 -
4.1.3. 皮膚細胞株暴露UVB後胞內 ROS的變化 - 51 -
4.1.4. 皮膚細胞抗氧化酵素變化情形 - 51 -
4.2. 探討抗氧化物對不同細胞株的保護能力模式建立方法實驗 - 51 -
4.2.1. 不同抗氧化物對皮膚細胞株的毒性測試 - 51 -
4.2.2. 探討抗氧化物對不同細胞株的保護能力模式建立 - 53 -
4.2.3. 抗氧化物對皮膚細胞的保護效果 - 54 -
4.2.4. 阿魏酸木寡糖在人類角質層細胞受UVB 60 mJ/cm2 暴露後，細胞內清除ROS 和超氧陰離子 - 54 -
第五章 結 論 - 74 -
參考文獻 - 76 -
中文部份 - 76 -
附錄 一 - 84 -
附錄 二 - 86 -
圖目錄
圖3-1. 紫外光 / 可見光光譜分析儀（USB 4000）與配件。 - 39 -
圖3-2. 紫外線暴露細胞模式。 - 40 -
圖3-3. 手持式紫外線燈管。 - 41 -
圖3-4. 紫外線暴露細胞實際圖。 - 41 -
圖3-5. ROS assay 流程圖。 - 42 -
圖3-6. Dihydroethidium assay 流程圖。 - 43 -
圖3-7. LDH assay 流程圖。 - 44 -
圖3-8. MTT assay 流程圖。 - 45 -
圖3-9. 過氧化氫酶分析方法流程圖。 - 46 -
圖3-10. 蛋白質濃度測定方法流程圖。 - 47 -
圖3-11. 蛋白質測定檢量線。 - 48 -
圖4-1. USB 4000紫外線可見光光譜分析系統量測手持式紫外線燈管在5~30公分下劑量功率圖。 - 56 -
圖4-2. UVB燈管在高度5公分所量測的功率圖譜。 - 56 -
圖4-3. 老鼠角質層細胞暴露不同UVB劑量下，LDH在不同時間的變化情形。- 57 -
圖4-4. 老鼠角質層細胞暴露不同UVB劑量下，細胞存活率在不同時間的變化情形。- 57 -
圖4-5. UVB對老鼠角質層細胞的有效劑量（ED 50）。 - 58 -
圖4-6. 人類角質層細胞暴露不同UVB劑量下，細胞毒性在不同時間的變化。 - 58 -
圖4-7. 人類角質層細胞暴露不同UVB劑量下，細胞存活率在不同時間的變化。 - 59 -
圖4-8. 老鼠角質層細胞暴露UVB 60 mJ/cm2，胞內不同時間點ROS的變化。 - 60 -
圖4-9. 老鼠角質層細胞暴露UVB 60 mJ/cm2，胞內ROS和Catalase變化情形。 - 61 -
圖4-10. 三種抗氧化物1 mg/ml對老鼠角質層細胞的存活率測試。 - 62 -
圖4-11. 三種抗氧化物1 mg/ml對老鼠角質層細胞的毒性測試。 - 62 -
圖4-12. 三種抗氧化物0.5 mg/ml對人類角質層細胞的毒性測試。 - 63 -
圖4-13. 三種抗氧化物0.5 mg/ml對人類角質層細胞的存活率測試。 - 63 -
圖4-14. 不同抗氧化物對老鼠角質層細胞株暴露UVB 60 mJ/cm2 的光保護效果。 - 64 -
圖4-15. 不同抗氧化物對人類角質層細胞株暴露UVB 60 mJ/cm2 的光保護效果。 - 65 -
圖4-16. 抗氧化物OFA對人類角質層細胞株暴露UVB 60 mJ/cm2 的清除ROS 效果。 - 66 -
圖4-17. 抗氧化物OFA對人類角質層細胞株暴露UVB 60 mJ/cm2 的清除超氧陰離子效果。 - 67 -
附錄圖1. 不同濃度的阿魏酸對老鼠角質層細胞存活率測試。 - 86 -
附錄圖2. 不同濃度的阿魏酸木寡糖對老鼠角質層細胞存活率測試。 - 86 -
附錄圖3. 不同濃度的維生素C對老鼠角質層細胞存活率測試。 - 87 -
附錄圖4. 不同濃度的阿魏酸對老鼠角質層細胞的毒性測定結果。 - 87 -
附錄圖5. 不同濃度的阿魏酸木寡糖對老鼠角質層細胞的毒性測定結果。 - 88 -
附錄圖6. 不同濃度的維生素C對老鼠角質層細的毒性測定結果。 - 88 -
附錄圖7. 不同濃度的阿魏酸對人類角質層細胞的存活率測試。。。。 - 89 -
附錄圖8. 不同濃度的阿魏酸木寡糖對人類角質層細胞的存活率測試。 - 89 -
附錄圖9. 不同濃度的維生素 C對人類角質層細胞的存活率測試。。 - 90 -
附錄圖10. 不同濃度的阿魏酸對人類角質層細胞的LDH毒性測定結果。 - 90 -
附錄圖11. 不同的濃度的阿魏酸木寡糖對人類角質層細胞的LDH毒性測定結果。 - 91 -
附錄圖12. 不同濃度的維生素 C對人類角質層細胞的LDH毒性測定結果。 - 91 -
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Yuan XP, Wang J, Yao H. (2006).Production of feruloyl oligosaccharides from wheat bran insoluble dietary fibre by xylanases from Bacillus subtilis. Food Chemistry, 95：484-492.[[abstract]]目前文獻中針對皮膚暴露紫外線輻射的研究，對於測量紫外線劑量的方式，常以直讀式儀器進行量測其功率，但此種方式對於評估皮膚暴露紫外線的影響將受到限制。因此本研究為利用USB 4000 紫外線/可見光光譜分析儀，量測紫外燈管之輻射劑量，以不同的紫外線輻射劑量，暴露於老鼠及人類皮膚細胞株，誘發細胞產生氧化壓力傷害；並獲得紫外線對細胞暴露的半效應劑量（ED50），進而以此劑量作為探討抗氧化物對皮膚細胞株的保護能力效應之應用。
兩種皮膚細胞株暴露UVB 60 mJ/cm2 劑量後，胞內活性氧族群（ROS assay）與胞外乳酸脫氫酶（LDH assay）明顯的增加，而細胞存活率指標值 （MTT assay） 降低，顯示此劑量能顯著的造成細胞傷害。以此模式評估不同抗氧化物對細胞暴露UVB的保護效果，結果顯示阿魏酸、阿魏酸木寡醣與維生素C，對老鼠皮膚細胞（1 mg/ml）和人類角質層細胞（0.5、1 mg/ml）均具有保護效果，可以減少經由UVB 所產生的氧化壓力傷害。
本研究之紫外線暴露模式改善了直讀式量測儀器的偵測缺失，可以提供光源光譜功率圖和光波長範圍，對燈源的特性資訊做進一步的確認，細胞暴露UVB模式的建立亦可以應用於評估抗氧化物質的效果。
關鍵字：皮膚細胞、光譜分析儀、紫外線、抗氧化物、氧化壓力
The utilization of Radiometer to measure ultraviolet radiation in skin exposure assessment is limited since the lack of spectral irradiance and full wavelength scanning. An USB 4000 Miniature Fiber Optic Spectrometer was alternative used to measure the UVR emission and exposure dose from UV lamps. Following the measuring method, UVB induced oxidative stress in keratinocytes cell lines, xb-2 (mouse) and HaCaT (human), were establishes. This model was further applied to assess the protective effect of certain antioxidant agents.
After UVB 60 mJ/cm2 exposure and then incubation 24 hr was found as the half efficiency does (ED50) in preliminary test. Both mouse and human keratinocyte cell lines showed the cellular ROS (reactive oxygen species), extracellular LDH (lactate dehydrogenase) increased, and in coincidence with the cellular survival (MTT test) decrease under the condition of ED50. The effects of antioxidant agents such as ferulic acid, feruloyl oligosaccharides, and Vitamin C showed protective capacity in this model.
Based on the results from this study, the UVB exposure model could more accurately provides the information of spectrophotographies and UVB dosage. Moreover, the UV exposure model was evidenced to apply in evaluating the protective effect of antioxidant agents in keratinocytes.
Key word：keratinocyte、ultraviolet radiation、antioxidant agents oxidative stress、Spectroradiometer syste
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