Chung Hwa University of Medical Technology
Chung Hwa University of Medical Technology RepositoryNot a member yet
1453 research outputs found
Sort by
Use the Coefficient of Determination Properly to Assess the Agreement of Two Sets of Measurements
No full text
Comparison and analysis of Clinical Chemistry and Hematology are studied before and after Anesthetization with Local Anesthetics to Crucian carp,Carassius auratus auratus
No full textThe study on Organization Climate and Organization Commitment in Golf Recreational Business Staffs-Tainan City and Tainan County for Example
No full textStudy of Differential Pressure in Airborne Infection Isolation Room Due to Doorway and Leakage
Get PDF[[abstract]]負壓隔離病房需維持病室內呈現負壓且確保氣密，以防止致病性微生物向外逸散。當病房內除門縫開口外仍存在其他洩漏源時，對於負壓測值之影響有待深入探討。本研究使用可改變體積之組合式環境模擬暴露艙並配合變頻馬達調整進排氣風量，以探討門縫開口與洩漏開口對負壓測值之影響。設定艙體體積介於45.38 m^3 ~ 28.47 m^3之間並調整進排氣風量(將進氣量當作計算基準以推算出1.2倍的排氣量)，以得到所需的每小時換氣次數(以下簡稱ACH)分別為4、8、12及16次，可調整的門縫開口高度介於0.2 cm ~ 1.2 cm之間且門縫寬度固定為100 cm，結合不同洩漏開口面積(4、25及100 cm^2)條件設定，評估在無計畫性洩漏(僅有門縫開口)與計畫性洩漏(門縫開口與洩漏開口)兩種狀況下，門縫開口高度與總洩漏面積對負壓測值之影響。根據實驗所得的數據分析發現，不同的體積下，ACH越大其負壓測值越大，隨著門縫開口高度增加其負壓測值逐漸減小，此外在本研究所採用的體積範圍內並未發現負壓測值因為洩漏開口位置的不同而改變，故可由門縫面積與洩漏面積兩者計算總洩漏面積，評估其在不同的ACH次數下總洩漏面積對負壓測值之影響。本研究結果發現，ACH為4 次且總洩漏面積大於或等於25 cm^2時，則無法達到我國對於負壓隔離病房之建議值(≧ 8 Pa)。最後，根據本研究數據已發展出壓差預測模式，可由現場進排氣風量差及門縫開口面積的實測值推估其負壓測值之大小，提供設計建造判定是否有其他洩漏來源以確保運作有效性。
Negative pressure and air-tightness are required for airborne infection isolation room (AIIR) to prevent the escape of microorganisms. The effect of unexpected leakage in AIIR on decreasing negative pressure differential need to be investigated. In this research, a volume-adjustable chamber with independent supply and exhaust ventilation system which simulate airborne infection isolation room without anteroom was adopted to study the pressure differential caused by doorway and leakage. The ventilation system provides 20% greater exhaust flow-rate than supply air to meet 4, 8, 12 and 16 air change per hour (ACH) of the chamber with the volume ranging between 28.47 m^3 and 45.38 m^3. By changing the adjustable height (0.2 ~ 1.2 cm) of doorway which has 100 cm width, the value of negative pressure was measured to develop the correlation of doorway size and pressure differential. Similar experiment was also conducted under three different designed leakage areas (4, 25 and 100 cm^2) to study the effect of leakage area on pressure differential.It was found that pressure differential increases as ACH increases, but decreases with decreasing height of doorway and leakage area for different chamber volumes. No significant pressure differential was observed for different leakage positions. For ACH equals to 4, the pressure differential can not reach the recommended standard of pressure differential (≧8 Pa) when leakage area is greater than 25 cm^2. By using the experimental data, a predicting model was developed to estimate pressure differential based on the information of flow-rate, doorway and leakage area. It provides diagnosis of leakage sources and ensures operational effectiveness of AIIR
Elucidation of the mechanism that BMP-2 suppresses TGF-β1-induced renal cellular fibrosis
No full text[[abstract]]上皮-間質轉換EMT ( epithelial -mesenchymal transition ) 在胚胎發育上是一個不可或缺的過程。但是在某些晚期的慢性腎疾病中，腎臟纖維化的間質纖維母細胞確是源自於一群腎小管的上皮細胞經歷了一個EMT的過程而來。目前已經知道第一型乙型轉型生長因子TGF-β1 ( transforming growth factor-beta1 ) 能夠在腎臟纖維化的過程中誘導細胞纖維化並且促進EMT的發生。儘管如此，上皮細胞可塑性的能力也同時受到TGF-β1的相關蛋白第二型骨髓生長因子BMP-2 ( bone morphogenetic protein-2 ) 所調節。我們先前的研究發現正常大鼠腎間質纖維母細胞 ( NRK-49F ) 短暫刺激BMP-2 ( 24hr ) 能對抗TGF-β1誘發腎臟細胞纖維化的現象。在這份研究中，我們將探討在正常大鼠腎間質纖維母細胞中BMP-2、TGF-β1和EMT的相互調控機制。培養NRK-49F於TGF-β1 ( 10 ng/mL ) 會誘導EMT的進行，使得α-SMA、 Fibronectin和Snail蛋白表現增加以及E-cadherin蛋白的降低而逐漸失去上皮細胞的型態。若共同刺激TGF-β1 ( 10 ng/mL ) 和BMP-2 ( 10、50、250 ng/mL ) 的劑量24小時則可以逆轉TGF-β1誘導EMT的進行，並且再度重新表現E-cadherin蛋白並且降低α-SMA、Fibronectin和Snail蛋白。這些數據表示BMP-2抑制TGF-β1誘導正常大鼠腎間質纖維母細胞的纖維化會經由調節Snail表現以及EMT的過程。
EMT ( epithelial- mesenchymal transition ) is an integral process during development. But some later phase of chronic kidney disease, the interstitial fibroblasts actually originate from tubular epithelial cells via the EMT in renal fibrogenesis. Transforming growth factor beta ( TGF-β1 ) has been known as the main cytokine involved in the pathogenesis of renal fibrogenesis and also capable of initiating and completing the entire EMT course. Although, epithelial plasticity can also be regulated by a member of the TGF-β1 superfamily bone morphogenetic protein-2 ( BMP-2 ). Our previous study showed that short-term treatment ( 24 hr ) of BMP-2 in NRK-49F ( rat fibroblast ) cells could antagonize TGF-β1 -induced fibrogenesis. In this study, We investigate the interactions of BMP-2, TGF-β1 and EMT process in NRK-49F. Cultured NRK-49F cells were treated with TGF-β1 ( 10 ng/mL ) for 24 hours could be induced EMT, in association with increased α-SMA, Fibronectin, Snail expression, decreased E-cadherin expression and loss of epithelial morphology. A combination of TGF-β1 ( 10 ng/mL ) in BMP-2 dose-dependent manner ( 10, 50, 250 ng/mL ) for 24 hours could be reversed TGF-β1-induced EMT, in association with reexpression of endogenous E-cadherin and degrade α-SMA, Fibronectin and Snail. These data suggest that BMP-2 attenuates TGF-β1-induced renal cell fibrosis could be through mediated Snail and EMT process
The prevention effect of Hylocereus polyrhizus extract on the brain peroxidation injury with aged-loss of spontaneous locomotor activity in aged mice
No full text[[abstract]]由於邁向高齡化社會，老化所造成的退化性疾病，成為重要的醫療問題。單胺氧化酶(Monoamine oxidase, MAO)及行為的缺乏在人體老化及其他神經相關疾病(如：帕金森氏症、憂鬱症)扮演重要角色。因此本實驗探討紅龍果萃取物對MAO活性的影響外，近年的研究得知行為能力的降低也是老化過程中重要的生理指標，故本研究將進一步探討補充紅龍果萃取物對腦部氧化傷害及自發性活動能力影響之相關性。之前研究也提及紅龍果的多酚化合物可抑制老齡小鼠腦部單胺氧化酶活性及過氧化傷害。本實驗採用BALB/c雌鼠，依體重隨機分成三組，分別餵食不同實驗飼料：年輕控制組(YC)、老齡控制組(OC、H)10g extract/kg diet，共飼養14週，每4週進行自發性活動能力試驗，犧牲後測定其腦部及肝臟MAO活性、過氧化指標(TBARS、protein-carbonyl)及抗氧化酵素(catalase、SOD、GPx)，非酵素性抗氧化指標(GSH、Vit. C)，血清檢測總抗氧化能力(ORAC)，並進一步探討對腦部MAO蛋白質表現之影響。結果顯示YC組年輕小鼠自發性活動能力隨著時間增加而略為降低，且YC組活動能力顯著高於二組老齡組(OC、H)。H組在第8與第12週活動能力達到高峰，且顯著高於餵食前之活動力。另外，H組腦中單胺氧化酶活性顯著低於OC組且與YC組相當，進一步分析MAO-B 蛋白質表現量，三組間則無顯著差異，顯示紅龍果萃取物的補充是影響在MAO-B酵素活性，而不是蛋白質的表現。各組腦部TBARS、protein carbonyl指標分析，結果顯示OC組明顯高於YC組，H組則回復至與YC組相同，顯示飲食補充紅龍果萃取物補充可改善腦部的氧化損傷。以ORAC法 (Oxygen-radical absorbance capacity)測定血漿總抗氧能力，H組總抗氧化能力顯著高於OC組並與YC組相當，顯示紅龍萃取物補充時間的延長，可反應體內總抗氧化能力的提升。而酵素性抗氧化活性Catalase 則OC組高於YC組，H組則回復至與YC相當。SOD活性分析，腦部及肝臟OC組高於YC組，H組則回復至與YC相當，顯示紅龍果萃取物的添加可提升老齡小鼠SOD活性。GPx的結果顯示腦部GPx不受飲食影響。而紅龍果萃取物的添加肝臟GPx活性有顯著的提升。非酵素性抗氧化物含量GSH，腦部及肝臟GSH含量OC組顯著低於YC組，而紅龍果萃取物補充可提升GSH活性。腦部及肝臟Vit. C含量為OC組顯著低於YC組，H組則回至與YC組相當。顯示老齡小鼠紅龍果萃取物的添加可防禦自由基的攻擊，改善體內的抗氧化情形。將上述指標都與自發性活動能力進行相關性分析，其分析數據顯示protein carbonyl含量與MAO活性改變與老齡小鼠自發性活動能力降低有關，而腦部TBARS含量與自發性活動能力則較關聯性較低，在抗氧化系統GPx、SOD、Catalase活性、Vit.C及GSH與老齡小鼠自發性活能的降低則顯著相關。上述顯示推測老齡小鼠自發性活動能力的降低可能為透過抗氧化系統的改善與氧化損傷的防禦來改善此現象的發生。紅龍果萃取物補充可有效預防因老化所導致的腦部抗氧化系統衰退及氧化損傷，藉此提高老齡小鼠的自發性活動能力。結果有助於預防神經退化性疾病的保健食品開發之重要依據。
As the move towards an aging society, the aging caused by degenerative diseases, and become an important medical problem. The enzyme monoamine oxidase ( MAO) and the behavior defects is closely related to neurodegenration disorders such as Parkinson's disease, depression, of play an important role. This study investigated the Hylocereus polyrhizus extract on MAO enzyme activity, the recent studies that behavior reduce is an important physiological indicators,of the aging process. experimental purpose was to further explore the Hylocereus polyrhizus extract supplement for brain oxidation impact injury and spontaneous locomotor activity correlation. The present study, BALB/c female mice were randomly divided into three groups according to body weight, were fed different experimental diets: the young control group (YC), aging control group (OC, H) 10 g extract / kg diet, raising a total of 14 weeks, every 4 weeks of spontaneous activity test, Brain and liver TBARS, protein carbonyle, MAO activity, antioxidative enzyme activity and antioxidants of mice fed control (YC, OC) and Hylocereus supplement (H) diet for 14 weeks.The results demonstrated spotonas locomotory activity increase and decreased over time with YC group. The H group spotonas locomotory activity level in 8 weeks and 12 weeks was peakaging.In addition th MAO activity level is about the same as in YC group compared with H group.Age-related change of thiobarbituric acid reactive substances (TBARS) and protein carbonyl level and the oxygen radical absorbance capacity (ORAC) in plasma and antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase activity or non-enzymatic antioxidants vitamin C (Vit.C) and glutamic acid (GSH) concentrations in brain and liver were also observed.The results demonstrated TBARS and protein carbonyl concentrations significantly increased in OC group compared with YC group and H group is about the same as in YC group in brain.The Hylocereus polyrhizus extract supplement can prerevent oxidation impact injury in brain.These oxygen radical absorbance capacity (ORAC) test significantly increase in H group compared with OC group. and H group is about the same as in YC group in plasma. In addition antioxidant ezymes SOD activity and catalase activity significantly increase in OC group compared with YC group in brain and H group is about the same as in YC group.These results demonstrated GPx activity significantly increase in H group compared with OC group in liver. The non-enzymatic antioxidants GSH, Vit. C concentrations significantly increase in OC group compared with YC group in brain and liver and Hylocereus polyrhizus extract supplement can increase GSH, Vit.C concentrations in brain and liver. However Hylocereus polyrhizus extract can prevent age-related change of antioxidant system. and spontaneous locomotor activity.The results demonstrated significantly reduce spontaneous locomotor activity in mouse wity protein carbonyl concentrations and MAO activity have high level correlation in brain.Summary this study speculated reulated through prevention effect of Hylocereus polyrhizus extract on the brain peroxidation injury with aged-loss of spontaneous locomotor activity in aged mice.The study above can proved for the bases of food selections neurodegenerative prevention
Studies of antibacterial and antioxidant activity of essential oil and water extracts of peel of Citrus grandis (L.) Osbeck
No full textAndrographolide down-regulates hypoxia-inducible factor-1α in human lung cancer A549 cells
No full text口試委員會審定書
授權書
中文摘要……………………………………………………………………………i
英文摘要…………………………………………………………………iii
誌 謝……………………………………………………………………v
目 錄……………………………………………………………………vii
第一章 文獻資料回顧
第一節 肺 癌………………………………………………………………………1
第二節 低氧誘導因子基本介紹……………………………………………3
第三節 穿心蓮……………………………………………………………………9
第四節 穿心蓮內酯…………………………………………………………10
第二章 研究目的…………………………………………………………………13
第三章 研究架構………………………………………………………………14
第四章 實驗材料與方法
第一節 實驗材料與實驗器材
1. 藥品試劑………………………………………………………………15
2. 實驗儀器………………………………………………………………23
第二節 實驗方法
1. 細胞培養………………………………………………………………24
2. 蛋白質抽取……………………………………………………………24
3. 西方墨點法……………………………………………………………25
4. 即時定量 PCR…………………………………………………………28
5. 免疫沉澱法……………………………………………………………29
6. 膠體電泳位移分析法…………………………………………………29
第五章 實驗結果
第一節 穿心蓮內酯對於肺癌細胞A549中HIF-1α之影響……………33
第二節 穿心蓮內酯對於肺癌細胞A549中調控HIF-1α相關因子之
影響.................................................33
第三節 穿心蓮內酯促進肺癌細胞A549中HIF-1α結合Ubiquitin的
表現增加..............................................34
第四節 穿心蓮內酯調控肺癌細胞A549中HIF-1α及相關因子之轉
錄活性影響…………………………………………………………35
第五節 穿心蓮內酯調控肺癌細胞A549中血管新生因子(VEGF) 蛋
白質和mRNA之表現………………………………………36
第六章 討論與結論………………………………………………………………37
參考文獻………………………………………………………………………42
圖 表…………………………………………………………………………………52
附 錄…………………………………………………………………………………64
參考文獻
Aggarwal, B.B., Banerjee, S., Bharadwaj, U., Sung, B., Shishodia, S., and Sethi, G. (2007). Curcumin induces the degradation of cyclin E expression through ubiquitin-dependent pathway and up-regulates cyclin-dependent kinase inhibitors p21 and p27 in multiple human tumor cell lines. Biochem Pharmacol, 73, 1024-1032.
Akbarsha, M.A., and Murugaian, P. (2000). Aspects of the male reproductive toxicity/male antifertility property of andrographolide in albino rats: effect on the testis and the cauda epididymidal spermatozoa. Phytother Res, 14, 432-435.
Bardos, J.I., and Ashcroft, M. (2005). Negative and positive regulation of HIF-1: a complex network. Biochim Biophys Acta, 1755, 107-120.
Batkhuu, J., Hattori, K., Takano, F., Fushiya, S., Oshiman, K., and Fujimiya, Y. (2002). Suppression of NO production in activated macrophages in vitro and ex vivo by neoandrographolide isolated from Andrographis paniculata. Biol Pharm Bull, 25, 1169-1174.
Brahimi-Horn, C., Mazure, N., and Pouyssegur, J. (2005). Signalling via the hypoxia-inducible factor-1alpha requires multiple posttranslational modifications. Cell Signal, 17, 1-9.
Burgos, R.A., Seguel, K., Perez, M., Meneses, A., Ortega, M., Guarda, M.I., Loaiza, A., and Hancke, J.L. (2005). Andrographolide inhibits IFN-gamma and IL-2 cytokine production and protects against cell apoptosis. Planta Med, 71, 429-434.
Calabrese, C., Berman, S.H., Babish, J.G., Ma, X., Shinto, L., Dorr, M., Wells, K., Wenner, C.A., and Standish, L.J. (2000). A phase I trial of andrographolide in HIV positive patients and normal volunteers. Phytother Res, 14, 333-338.
Cheung, H.Y., Cheung, S.H., Li, J., Cheung, C.S., Lai, W.P., Fong, W.F., and Leung, F.M. (2005). Andrographolide isolated from Andrographis paniculata induces cell cycle arrest and mitochondrial-mediated apoptosis in human leukemic HL-60 cells. Planta Med, 71, 1106-1111.
Chiou, W.F., Chen, C.F., and Lin, J.J. (2000). Mechanisms of suppression of inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells by andrographolide. Br J Pharmacol, 129, 1553-1560.
Choi, H.J., Song, B.J., Gong, Y.D., Gwak, W.J., and Soh, Y. (2008). Rapid degradation of hypoxia-inducible factor-1alpha by KRH102053, a new activator of prolyl hydroxylase 2. Br J Pharmacol, 154, 114-125.
Du, K.M., Chen, G.Q., and Chen, Z. (2004). Regulation of hypoxia-inducible factor-1alpha expression. Ai Zheng, 23, 1098-1102.
Fedele, A.O., Whitelaw, M.L., and Peet, D.J. (2002). Regulation of gene expression by the hypoxia-inducible factors. Mol Interv, 2, 229-243.
Folkman, J. (2002). Role of angiogenesis in tumor growth and metastasis. Semin Oncol, 29, 15-18.
Forsythe, J.A., Jiang, B.H., Iyer, N.V., Agani, F., Leung, S.W., Koos, R.D., and Semenza, G.L. (1996). Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol, 16, 4604-4613.
Groulx, I., and Lee, S. (2002). Oxygen-dependent ubiquitination and degradation of hypoxia-inducible factor requires nuclear-cytoplasmic trafficking of the von Hippel-Lindau tumor suppressor protein. Mol Cell Biol, 22, 5319-5336.
Haddad, J.J. (2004). Oxygen sensing and oxidant/redox-related pathways. Biochem Biophys Res Commun, 316, 969-977.
Handa, S.S., and Sharma, A. (1990). Hepatoprotective activity of andrographolide from Andrographis paniculata against carbonte- trachloride. Indian J Med Res, 92, 276-283.
Hanahan D., and Weinberg RA. (2000) The hallmarks of cancer. Cell,100,57-70.
Hidalgo, M.A., Romero, A., Figueroa, J., Cortes, P., Concha, II, Hancke, J.L., and Burgos, R.A. (2005). Andrographolide interferes with binding of nuclear factor-kappaB to DNA in HL-60-derived neutrophilic cells. Br J Pharmacol, 144, 680-686.
Jada, S.R., Hamzah, A.S., Lajis, N.H., Saad, M.S., Stevens, M.F., and Stanslas, J. (2006). Semisynthesis and cytotoxic activities of andrographolide analogues. J Enzyme Inhib Med Chem, 21, 145-155.
Jeong, J.W., Bae, M.K., Ahn, M.Y., Kim, S.H., Sohn, T.K., Bae, M.H., Yoo, M.A., Song, E.J., Lee, K.J., and Kim, K.W. (2002). Regulation and destabilization of HIF-1alpha by ARD1-mediated acetylation. Cell, 111, 709-720.
Kaelin, W.G., Jr. (2008). The von Hippel-Lindau tumour suppressor protein: O2 sensing and cancer. Nat Rev Cancer, 8, 865-873.
Kelly, B.D., Hackett, S.F., Hirota, K., Oshima, Y., Cai, Z., Berg-Dixon, S., Rowan, A., Yan, Z., Campochiaro, P.A., and Semenza, G.L. (2003). Cell type-specific regulation of angiogenic growth factor gene expression and induction of angiogenesis in nonischemic tissue by a constitutively active form of hypoxia-inducible factor 1. Circ Res, 93, 1074-1081.
Kim, T.G., Hwi, K.K., and Hung, C.S. (2005). Morphological and biochemical changes of andrographolide-induced cell death in human prostatic adenocarcinoma PC-3 cells. In Vivo, 19, 551-557.
Konishi, T., Huang, C.L., Adachi, M., Taki, T., Inufusa, H., Kodama, K., Kohno, N., and Miyake, M. (2000). The K-ras gene regulates vascular endothelial growth factor gene expression in non-small cell lung cancers. Int J Oncol, 16, 501-511.
Lee, Y.C., Lin, H.H., Hsu, C.H., Wang, C.J., Chiang, T.A., and Chen, J.H. (2010). Inhibitory effects of andrographolide on migration and invasion in human non-small cell lung cancer A549 cells via down-regulation of PI3K/Akt signaling pathway. Eur J Pharmacol, 632, 23-32.
Lin, T.P., Chen, S.Y., Duh, P.D., Chang, L.K., and Liu, Y.N. (2008). Inhibition of the epstein-barr virus lytic cycle by andrographolide. Biol Pharm Bull, 31, 2018-2023.
Liu, L.Z., Fang, J., Zhou, Q., Hu, X., Shi, X., and Jiang, B.H. (2005). Apigenin inhibits expression of vascular endothelial growth factor and angiogenesis in human lung cancer cells: implication of chemoprevention of lung cancer. Mol Pharmacol, 68, 635-643.
Mazure, N.M., Brahimi-Horn, M.C., Berta, M.A., Benizri, E., Bilton, R.L., Dayan, F., Ginouves, A., Berra, E., and Pouyssegur, J. (2004). HIF-1: master and commander of the hypoxic world. A pharmacological approach to its regulation by siRNAs. Biochem Pharmacol, 68, 971-980.
Mehrotra, R., Rawat, S., Kulshreshtha, D.K., Patnaik, G.K., and Dhawan, B.N. (1990). In vitro studies on the effect of certain natural products against hepatitis B virus. Indian J Med Res, 92, 133-138.
Merk J., Leschber G. (2009). Adjuvant therapy for lung cancer. Minerva Chirurgica, 64, 599-609.
Rajagopal, S., Kumar, R.A., Deevi, D.S., Satyanarayana, C., and Rajagopalan, R. (2003). Andrographolide, a potential cancer therapeutic agent isolated from Andrographis paniculata. J Exp Ther Oncol, 3, 147-158.
Rankin, E.B., and Giaccia, A.J. (2008). The role of hypoxia-inducible factors in tumorigenesis. Cell Death Differ, 15, 678-685.
Ravi, R., Mookerjee, B., Bhujwalla, Z.M., Sutter, C.H., Artemov, D., Zeng, Q., Dillehay, L.E., Madan, A., Semenza, G.L., and Bedi, A. (2000). Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha. Genes Dev, 14, 34-44.
Ruas, J.L., Poellinger, L., and Pereira, T. (2005). Role of CBP in regulating HIF-1-mediated activation of transcription. J Cell Sci, 118, 301-311.
Sang, N., Stiehl, D.P., Bohensky, J., Leshchinsky, I., Srinivas, V., and Caro, J. (2003). MAPK signaling up-regulates the activity of hypoxia-inducible factors by its effects on p300. J Biol Chem, 278, 14013-14019.
Satyanarayana, C., Deevi, D.S., Rajagopalan, R., Srinivas, N., and Rajagopal, S. (2004). DRF 3188 a novel semi-synthetic analog of andrographolide: cellular response to MCF 7 breast cancer cells. BMC Cancer, 4, 26.
Semenza, G.L. (2003). Targeting HIF-1 for cancer therapy. Nat Rev Cancer, 3, 721-732.
Semenza, G.L. (2007). Evaluation of HIF-1 inhibitors as anticancer agents. Drug Discov Today, 12, 853-859.
Shen, Y.C., Chen, C.F., and Chiou, W.F. (2000). Suppression of rat neutrophil reactive oxygen species production and adhesion by the diterpenoid lactone andrographolide. Planta Med, 66, 314-317.
Shi, M.D., Lin, H.H., Chiang, T.A., Tsai, L.Y., Tsai, S.M., Lee, Y.C., and Chen, J.H. (2009). Andrographolide could inhibit human colorectal carcinoma Lovo cells migration and invasion via down-regulation of MMP-7 expression. Chem Biol Interact, 180, 344-352.
Shi, M.D., Lin, H.H., Lee, Y.C., Chao, J.K., Lin, R.A., and Chen, J.H. (2008). Inhibition of cell-cycle progression in human colorectal carcinoma Lovo cells by andrographolide. Chem Biol Interact, 174, 201-210.
Shima, D.T., Kuroki, M., Deutsch, U., Ng, Y.S., Adamis, A.P., and D'Amore, P.A. (1996). The mouse gene for vascular endothelial growth factor. Genomic structure, definition of the transcriptional unit, and characterization of transcriptional and post-transcriptional regulatory sequences. J Biol Chem, 271, 3877-3883.
Shukla, B., Visen, P.K., Patnaik, G.K., and Dhawan, B.N. (1992). Choleretic effect of andrographolide in rats and guinea pigs. Planta Med, 58, 146-149.
Singha, P.K., Roy, S., and Dey, S. (2007). Protective activity of andrographolide and arabinogalactan proteins from Andrographis paniculata Nees. against ethanol-induced toxicity in mice. J Ethnopharmacol, 111, 13-21.
Smith, T.G., Robbins, P.A., and Ratcliffe, P.J. (2008). The human side of hypoxia-inducible factor. Br J Haematol, 141, 325-334.
Stoeltzing, O., McCarty, M.F., Wey, J.S., Fan, F., Liu, W., Belcheva, A., Bucana, C.D., Semenza, G.L., and Ellis, L.M. (2004). Role of hypoxia-inducible factor 1alpha in gastric cancer cell growth, angiogenesis, and vessel maturation. J Natl Cancer Inst, 96, 946-956.
Tal, R., Shaish, A., Bangio, L., Peled, M., Breitbart, E., and Harats, D. (2008). Activation of C-transactivation domain is essential for optimal HIF-1 alpha-mediated transcriptional and angiogenic effects. Microvasc Res, 76, 1-6.
Tang, N., Wang, L., Esko, J., Giordano, F.J., Huang, Y., Gerber, H.P., Ferrara, N., and Johnson, R.S. (2004). Loss of HIF-1alpha in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis. Cancer Cell, 6, 485-495.
Thisoda, P., Rangkadilok, N., Pholphana, N., Worasuttayangkurn, L., Ruchirawat, S., and Satayavivad, J. (2006). Inhibitory effect of Andrographis paniculata extract and its active diterpenoids on platelet aggregation. Eur J Pharmacol, 553, 39-45.
Tsai, H.R., Yang, L.M., Tsai, W.J., and Chiou, W.F. (2004). Andrographolide acts through inhibition of ERK1/2 and Akt phosphorylation to suppress chemotactic migration. Eur J Pharmacol, 498, 45-52.
Visen, P.K., Shukla, B., Patnaik, G.K., and Dhawan, B.N. (1993). Andrographolide protects rat hepatocytes against paracetamol-induced damage. J Ethnopharmacol, 40, 131-136.
Wang, T., Liu, B., Zhang, W., Wilson, B., and Hong, J.S. (2004). Andrographolide reduces inflammation-mediated dopaminergic neurodegeneration in mesencephalic neuron-glia cultures by inhibiting microglial activation. J Pharmacol Exp Ther, 308, 975-983.
Wiart, C., Kumar, K., Yusof, M.Y., Hamimah, H., Fauzi, Z.M., and Sulaiman, M. (2005). Antiviral properties of ent-labdene diterpenes of Andrographis paniculata nees, inhibitors of herpes simplex virus type 1. Phytother Res, 19, 1069-1070.
Yeo, E.J., Chun, Y.S., and Park, J.W. (2004). New anticancer strategies targeting HIF-1. Biochem Pharmacol, 68, 1061-1069.
Yu, B.C., Hung, C.R., Chen, W.C., and Cheng, J.T. (2003). Antihyperglycemic effect of andrographolide in streptozotocin-induced diabetic rats. Planta Med, 69, 1075-1079.
Zagorska, A., and Dulak, J. (2004). HIF-1: the knowns and unknowns of hypoxia sensing. Acta Biochim Pol, 51, 563-585.
Zhang, Z., Jiang, J., Yu, P., Zeng, X., Larrick, J.W., and Wang, Y. (2009). Hypoglycemic and beta cell protective effects of andrographolide analogue for diabetes treatment. J Transl Med, 7, 62.[[abstract]]肺癌是全球性癌症致死率的第一位，且其發生率在許多國家都有逐漸增加的趨勢。肺癌轉移往往造成治療上的困難，也是導致肺癌治癒率低的主要原因。血管新生對於腫瘤生成和轉移來說扮演非常重要的角色，它可以供給腫瘤氧氣、營養和一些生長因子的需求。低氧誘導因子-1α ( HIF-1α ) 是一種調控體內氧平衡和細胞缺氧時反應的轉錄因子，它可調控下游血管內皮生長因子 ( VEGF ) 基因的活化，促進腫瘤的生長。穿心蓮內酯 ( Andrographolide ) 是由傳統中草藥穿心蓮所分離出的物質，為穿心蓮中的主要功效成分。過去我們研究發現，穿心蓮內酯具有抑制肺癌細胞生長與轉移的特定療效。因此，本實驗將探討穿心蓮內酯對於人類肺癌細胞A549的HIF-1α之調控機制，以及是否會干擾HIF-1α下游因子的表現進而影響血管新生的作用。經由西方墨點法 ( Western blotting ) 證實，穿心蓮內酯可減少細胞內HIF-1α 和VEGF的蛋白表現，進一步研究發現穿心蓮內酯可以促進hydroxy-HIF-1α ( Pro564 ) 蛋白增加並造成 HIF-1α 的蛋白分解，分析其中調控機制發現，促進 HIF-1α 分解的 PHD2 (prolyl hydroxylase)蛋白表現增加，而抑制 HIF-1α轉錄活性的 FIH-1 (Factor- inhibiting HIF-1)蛋白表現則不受影響。另外在RT-PCR的結果顯示，
ii
穿心蓮內酯似乎不影響HIF-1α mRNA的轉錄活性。此外，經由膠體電泳位移分析法 ( EMSA ) 的結果證實，穿心蓮內酯可以抑制 HIF-1α 蛋白結合上 VEGF promoter 的區域，因而抑制下游VEGF基因的表達。綜合以上的實驗結果顯示，穿心蓮內酯主要是藉由促進HIF-1α的分解，進而抑制其下游因子VEGF 的表現，此路徑可能與調控A549細胞進行血管新生及轉移有關。未來，穿心蓮內酯在肺癌化學治療或發展新的抗血管新生藥物上將有所裨益。
Lung cancer is the major cause of malignancy-related deaths worldwide, and its incidence is rising in many countries. It is notorious for its low cure rate, and the treatment of lung cancer is usually compromised by metastasis. In the processes of tumor growth, a rapidly growing tumor can induce lack of oxygen and develop a hypoxia environment. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that regulates genes involved in angiogenesis and metastasis. Andrographolide (Andro) is isolated from a traditional Chinese herbal medicine, Andrographis paniculate, and has been reported to possess a variety of pharmacological activities. Our previous study has been demonstrated that Andro can inhibit human lung cancer A549 cell migration and invasion which was associated with the inhibition of intracellular protein kinase cascades. In this study, we investigate whether Andro is an inhibitor of HIF-1α and its regulatory proteins, mediating downstream invasive or angiogenic signals. Using Western blotting and RT-PCR assay, it was revealed that the decreased expression of HIF-1α and vascular endothelial growth factor (VEGF), and the increase expressions of hydroxyl-HIF-1α (Pro564) and prolyl hydroxylase PHD2 in the Andro-treated A549 cells. The Andro-inhibited VEGF expression appeared to be a consequence of HIF-1α inactivation, because that it DNA binding activity was suppressed by Andro. In conclusion, we have elucidated the HIF-1α-VEGF signals was regulated by Andro. These results highlight the potential therapeutic effects of Andro, which may be developed as a chemotheraptic or an anti-angiogenesis agent in the future
Kinetic study of chlorine dioxide gas disinfection on bacillus Atrophaeus spores
Get PDF第一章 前言 1
1-1 研究背景 1
1-2 研究目的 1
第二章 文獻探討 3
2-1 二氧化氯的理化特性及其應用 3
2-2 二氧化氯進行飲用水水質處理的優點 5
2-3 空氣燻蒸消毒劑的類型 6
2-4 二氧化氯消毒作用機轉之研究 9
2-5 二氧化氯氣體對蔬菜水果中微生物殺菌效果的相關研究 10
2-6 二氧化氯的空氣消毒效果 14
2-7 二氧化氯暴露的健康危害 15
第三章 材料與方法 17
3-1 實驗設備器材及藥品 17
3-2 無菌稀釋液製備程序 21
3-3 二氧化氯穩定時間測試 22
3-4 二氧化氯燻蒸消毒實驗步驟 23
3-5 數據分析處理 24
第四章 結果與討論 29
4-1 二氧化氯溶液與燻蒸氣體濃度之關係 29
4-2 二氧化氯氣體濃度降解分析結果 30
4-3 燻蒸消毒作用劑量與孢子存活率分析 31
第五章 結論與建議 36
5-1 結論 36
5-2 建議 37
參考文獻 39
參考文獻
ATSDR(Agency for toxic substances and disease registry). (2004) Chorine dioxide and chlorite. Available from http:// www.atsdr.cdc.gov/tfacts160.html
Aydogan A. (2006) Application of Gaseous Disinfectants Ozone and Chlorine Dioxide for Inactivation of Bacillus Subtilis Spores. PhD Dissertation. San Diego State University, USA.
Du J, Han Y, Linton RH. (2002) Inactivation by chlorine dioxide gas (ClO2) of Listeria monocytogenes spotted onto different apple surfaces. Journal of Food Protection, 19, 481-490.
Du J, Han Y, Linton RH. (2003) Efficacy of chlorine dioxide gas in reducing Escherichia coli O157:H7 on apple surfaces. Food Microbiology 20:583-591.
Go´mez-Lo´pez VM, Rajkovic A, Ragaert P, Smigic N, Devlieghere F. (2009) Chlorine dioxide for minimally processed produce preservation: a review. Trends in Food Science & Technology 20, 17-26.
Han Y, Sherman DM, Linton RH, Nielsen SS, Nelson PE. (2000a) The effects of washing and chlorine dioxide gas on survival and attachment of Escherichia coli O157:H7 to green pepper surfaces. Food Microbiology 17:521-533.
Han Y, Linton RH, Nielsen SS, Nelson PE. (2000b) Inactivation of Escherichia coli O157:H7 on surface-uninjured and –injured green bell pepper (Capsicum annuum L.) by chlorine dioxide gas as demonstrated by confocal laser scanning microscopy. Food Microbiology 17:643-655.
Han Y, Linton RH, Nielsen SS, Nelson PE. (2001) Reduction of Listeria monocytogenes on green peppers (Capsicum annuum L.) by gaseous and aqueous chlorine dioxide and water washing and its growth at 7 ℃. Journal of Food Protection 64:1730-1738.
Jin R, Hu S, Zhang Y, Bo T. (2009) Concentration-dependence of the explosion characteristics of chlorine dioxide gas. Journal of Hazardous Materials 166:842-847.
Lee SY, Dancer GI, Chang SS, Rhee MS, Kang DH. (2006) Efficacy of chlorine dioxide gas against Alicyclobacillus acidoterrestris spores on apple surfaces. International Journal of Food Microbiology 108:364-368.
Luftman HS, Regits MA. (2008) B. Atrophaeus and G. Stearothermophilus Biological Indicators for Chlorine Dioxide Gas Decontamination. Applied Biosafety 13(3):143-57.
Luftman HS, Regits MA, Lorcheim P, Lorcheim K, and Paznek D. (2008) Validation Study for the Use of Chlorine Dioxide Gas as a Decontaminant for Biological Safety Cabinets. Applied Biosafety 13(4):199-212.
Mahmoud BSM, Linton RH. (2008) Inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce by chlorine dioxide gas. Food Microbiology 25:244-252.
Mahmoud BSM, Linton RH. (2008) Inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce by chlorine dioxide gas. Food Microbiology 25:244-252.
Mahmoud BSM, Bhagat AR, Linton RH. (2007) Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas. Food Microbiology 24:736-744.
Popa I, Hanson EJ, Todd ECD, Schilder AC, Ryser ET. (2007) Efficacy of chlorine dioxide gas sachets for enhancing the microbiological quality and safety of blueberries. Journal of Food Protection 70:2084-2088.
Rogers JV, Choi YW, Richter WR, Stone HJ, Taylor ML. (2008) Bacillus anthracis Spore Inactivation by Fumigant Decontamination. Applied Biosafety 13(2):89-98.
Rogers JV, Richter WR, Choi YW, Waugh JD, Taylor ML, Riggs KB, et al. (2006) Technology evaluation report: Evaluation of sporicidal decontamination technology, Sabre Technical Services chlorine dioxide gas generator. Available at: www.epa.gov/NHSRC/pubs/ erChlorineDioxide060506.pdf
Rogers JV, Sabourin CL, Taylor ML, Riggs K, Choi YW, Richter WR, Rudnicki DC, et al. (2004) Environmental technology verification report: CDG Research Corporation bench-scale chlorine dioxide gas: Solid generator. Available at: www.epa.gov/etv/pdfs/vrvs/11_vr_cdg.pdf
Singh N, Singh RK, Bhunia AK, Stroshine RL. (2002) Efficacy of chlorine dioxide, ozone, and thyme essential oil or a sequential washing in killing Escherichia coli O157:H7 on lettuce and baby carrots. Lebensmittel-Wissenschaft und-Technologie 35:720-729.
Sy KV, McWatters KH, Beuchat LR. (2005a) Efficacy of gaseous chlorine dioxide as a sanitizer for killing Salmonella, yeasts and molds on blueberries, strawberries, and raspberries. Journal of Food Protection 68:1165-1175.
Sy KV, Murray MB, Harrison MD, Beuchat LR. (2005b) Evaluation of gaseous chlorine dioxide as a sanitizer for killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and yeasts and molds on fresh and fresh-cut produce. Journal of Food Protection 68: 1176-1187.
WHO (World Health Organization) (2000) Environmental health criteria 216, disinfectants and disinfectant by-products. Available from http://www.inchem.org/documents/ehc/ehc/ehc216.htm#SectionNumber:8.1
Young SB, Setlow P. (2003) Mechanisms of killing of Bacillus subtilis spores by hypochlorite and chlorine dioxide. Journal of Applied Microbiology 95:54-67.
行政院勞工委員會(2003)勞工作業環境空氣中有害物容許濃度標準。行政院勞工委員會。
行政院環境保護署毒理資料庫(website: http://flora2.epa.gov.tw/toxicweb/toxicuc4/database1.asp)
李歸浦、夏會龍(2006)二氧化氯在食品滅菌消毒上的應用研究進展。食品科技9：21-25。
胡双啟、晉日亞(2007)氣體二氧化氯對水果的殺菌作用及其應用前景展望。中國安全科學學報 17(3)：153-155。
晉日亞、胡双啟(2008)氣體二氧化氯對蘋果表面細菌殺菌規律研究。食品科學 29(7)：109-112。
晉日亞、胡双啟(2008)氣體二氧化氯對蘋果表面細菌殺菌規律研究。食品科學 29(7)：109-112。
晉日亞、胡双啟、顧寧(2008)氣體二氧化氯對葡萄表面細菌殺菌規律研究。中國安全科學學報 29(7)：97-101。
書明肯、吳清平、王大鵬、吳軍林、張菊梅(2008)二氧化氯對脫氧核糖核苷三磷酸和質粒DNA的作用。微生物學通報35(8)：1224-1229。
張永良、鄭世英(2007)二氧化氯對不同類型房間空氣的消毒效果。環境與職業醫學24(3)：337-338。
張永良、鄭世英、智強(2007)二氧化氯對坑道空氣消毒的現場試驗。環境與健康雜誌24(4)：245-246。
張曉煜、吳清平、張菊梅、吳慧清、闕紹輝(2007)二氧化氯對大腸桿菌作用機理的研究。中國消毒學雜誌24(1):16-20。
張曉煜、鄧金花、黃靜、闕紹輝(2008)二不同活化條件對二氧化氯殺菌效果的影響。中國消毒學雜誌25(3):261-263。[[abstract]]本研究是利用經採用避光處理的壓克力材質製作的環境模擬暴露艙進行二氧化氯氣體對枯草桿菌孢子之空氣燻蒸消毒效能動力學探討。首先，利用市售的二氧化氯溶液經活化後加以稀釋並配製不同的濃度，再以超音波霧化器進行噴霧汽化後輸送進入暴露艙內，利用氣體檢知管進行空氣採樣分析以了解其濃度之時間變化情形，接著再利用經過高溫滅菌處理的棉線將含有枯草桿菌孢子(Bacillus Atrophaeus Spores)的生物指示孢菌片以固定間隔串接並均勻分散在艙內燻蒸消毒不同時間後取出放入TSB (Trypticase Soy Broth)培養液中觀察是否呈現渾濁的陽性反應率，之後再以無菌稀釋液進行十倍數逐步稀釋後以表面塗抹法完成二重覆樣本培養計數分析。結果顯示，二氧化氯溶液配製濃度不超過5000 mg/L時具有良好的線性關係，空氣中二氧化氯氣體濃度的半衰期約介於4-5小時之間，枯草桿菌孢子生物指示劑試紙存活之菌落生成數經過以對數尺度處理後，與二氧化氯氣體燻蒸消毒作用劑量間具有良好的線性相關。經計算其在100 ppm濃度之D值(Decimal reduction time)約為60分鐘，建議每次燻蒸消毒時間最好不超過2小時，根據二氧化氯氣體燻蒸消毒作用劑量可以推估要達到6 log量級之燻蒸循環次數，可提供規劃進行室內空氣消毒之參考依據。
The aim of this experiment is to study the inactivation kinetic of Bacillus atrophaeus spores with chlorine dioxide gas in a test chamber. Bacillus atrophaeus spores strips were treated with chlorine dioxide gas which was aerosolized by ultrasonic nebulizer from chlorine dioxide solution and introduced into test chamber at different concentrations in different time to determine the inactivated rate using enumeration and fractionation analysis of tryptic soy broth (TSB) and tryptic soy agar (TSA). The exposure dose was calculated by Harber’s rule based on chlorine dioxide gas concentration which was measured by detector tubes and total exposure time to establish D-value (Decimal reduction time) from inactivation curve. It was found that the half life of chlorine dioxide gas was less than 5 hr (300 min) at 20~25 ℃ under 90~95% relative humidity. The number of survived spores after exposure to chlorine dioxide gas followed an exponential decrease with increasing exposure concentration and contact time. The D-value of Bacillus atrophaeus spores treated with 100 ppm chlorine dioxide gas was about 60 minutes in this study. The results from this study suggest that gaseous chlorine dioxide should be a highly effective decontaminating agent for Bacillus atrophaeus spores within 2hr exposure time to determine the sterilization cycle achieving 6-log reduction