請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32123
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 潘子明 | |
dc.contributor.author | Yu-Ju Huang | en |
dc.contributor.author | 黃毓茹 | zh_TW |
dc.date.accessioned | 2021-06-13T03:32:45Z | - |
dc.date.available | 2009-08-01 | |
dc.date.copyright | 2006-08-01 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-26 | |
dc.identifier.citation | 日本厚生勞動省。(2001)。食品中毒關連情報. http://www.mhlw.go.jp/ topics/syokuchu/index.html.
王貴譽,張瑞烽。(1993)。大學微生物。國立編譯館。台北。台灣。中華民國 林彥嵐。(2000)。牛之糞材 Escherichia coli O157:H7 偵測流程之建立及其應用於乳牛場盛行率之調查。國立台灣大學獸醫學研究所碩士論文。台北。台灣。中華民國。 馬英萍。(2004)。台灣北部與東部乳牛場細菌藥物感受性及乳汁中藥物殘留調查。臺灣大學獸醫學研究所碩士論文。台北。台灣。中華民國.。 陳威戎,廖大修。(2003)。蛋白質體學的研究方法。In: 蛋白質體學. p:7-19。台北。台灣。中華民國 潘子明。(1996)。病源性大腸桿菌。健康世界。128: 45-49. 蔡文城。(1993)。實用臨床微生物診斷學。九州圖書文物有限公司。台北。台灣。中華民國 蔡宗佑。(2001)。O157:H7 型大腸桿菌之檢驗與分子分型之探討。國立台灣大學農業化學研究所碩士論文。台北。台灣。中華民國。 鄭景陽。(2000)。病源性大腸桿菌之致病性。食品科學與技術。32: 53-61. Adams, M. R. and Moss, M. O. (2000). Food Microbiology. Cambridge CB4 0WF, UK. Bakhtiar, R. and Nelson, R. W. (2001). Mass Spectrometry of the Proteome. Mol Pharmacol 60: 405-415. Beales, N. (2004). Adaptation of Microorganisms to Cold Temperatures, Weak Acid Preservatives, Low pH, and Osmotic Stress: A Review. Compr. Rev Food Sci Food Saf. 3: 1-20. Bearson, B. and W., Foster, J. W. (1997). Acid Stress Responses in Enterobacteria. FEMS Microbiol. Lett. 147: 173-180. Bernardini, M. L., Fontaine, A., Sansonetti, P. J. (1990) The Two-component Regulatory System ompR-envZ Controls the Virulence of Shigella flexneri. J Bacteriol. 172: 6274-6281. Calderwood, S. B., Acheson, D. W. K., Keusch, G. T., Barrett, T. J., Griffin, P. M., Strockbine, N. A., Swaminathan, B., Kaper, J. B., Levine, M. M., Kaplan, B. S., Karch, H., O'Brien, A. D., Obrig, T. G., Takeda, Y., Tarr, P. I. and Wachsmuth, I. K. (1996). Proposed New Nomenclature for SLT (VT) Family. ASM News 62: 118-119. Castanie-Cornet, M. P. and Foster, J. W. (2001). Escherichia coli Acid Resistance: cAMP Receptor Protein and a 20 bp cis-acting Sequence Control pH and Stationary Phase Expression of the gadA and gadBC Glutamate Decarboxylase Genes. Microbiol.147: 709-715. Cohen, M. B., Hawkins, J. A., Weckbach, L. S., Staneck, J. L., Levine, M. M. and Heck, J. E. (1993). Colonization by Enteroaggregative Escherichia coli in Travelers with and without Diarrhea. J of Clin Microbiol. 31: 351-353. Coia, J. E. (1998). Clinical, Microbiological and Epidemiological a Spects of Escherichia coli O157 Infection. FEMS Immuno Med Microbiol. 20:1-9 Diez-Gonzalez, F. and Russell, J. B. (1997). The Ability of Escherichia coli O157:H7 to Decrease Its Intracellular pH and Resist the Toxicity of Acetic Acid. Microbiol. 143: 1175-1180. Diez-Gonzalez, F., Callaway, T. R., Kizoulis, M. G. and Russell, J. B. (1998) Grain Feeding and the Dissemination of Acid-resistant Escherichia coli from Cattle. Science. 281:1666-1668 Donnenberg, M. S. and Kaper, J. B. (1992). Enteropathogenic Escherichia coli. . Infect Immun. 60: 3953-3961. Drasar, B. S. and Hill, M. J. (1974). Human Intestinal Flora. p:36-34. Academic Press, Ltd. London, United Kingdom. Endo, Y., Tsurugi, K., Yutsudo, T., Takeda, Y., Ogasawara, Y. and Igarashi, K. (1988). Site of Action of a Vero Toxin (Vt2) from Escherichia coli O157:H7 and of Shiga Toxin on Eucaryotic Ribosomes. Eur J Biochem. 171: 45-50. Evans, J., Wilson, A., Willshaw, G. A., Cheasty, T., Tompkins, D. S., Wheeler, J. G. and Smith, H. R. (2002). Vero Cytotoxin-producing Escherichia coli in a Study of Infectious Intestinal Disease in England. Clin Microbiol Infect 8: 183-186. Fenselau, C. 1997. MALDI-MS and Strategies for Protein Analysis. Anal Chem. 69: 661A-665A. Foster, J. W. (1991). Salmonella Acid Shock Proteins are Required for the Adaptive Acid Tolerance Response. J Bacteriol. 173: 6896-6902. Foster, J. W. (1993). The Acid Tolerance Response of Salmonella typhimurium Involves Transient Synthesis of Key Acid Shock Proteins. J Bacteriol. 175: 1981-1987. Foster, J. W. and Hall, H. K. (1990). Adaptive acidification tolerance response of salmonella typhimurium. J Bacteriol. 172: 771-778. Foster, J. W. and Moreno, M. (1999). Inducible Acid Tolerance Mechanisms in Enteric Bacteria. Novartis Found Symp. 221: 55-69. Foster, J. W. (2004) Escherichia coli Acid Resistance: Tales of an Amateur Acidophile. Nat Rev Microbiol. 2: 898-907 Goodson, M. and Rowbury, R. J. (1989). Habituation to Normally Lethal Acidity by Prior Growth of Escherichia coli at a Sub-Lethal pH Value. Lett Appl Microbiol. 8: 77-79. Gray, L. D. (1995). Escherichia, Salmonella, Shigella, and Yersinia. In: Manual of Clinical Microbiology. ASM Press. Washington, DC, USA. Guan, J. and Levin, R. E. (2002). Quantitative Detection of Escherichia coli O157:H7 in Ground Beef by the Polymerase Chain Reaction. Food Microbiol. 19: 159-165. Heengge-Aronis, R. (1999). Interplay of Global Regulators and Cell Physiology in the General Stress Response of Escherichia coli. Curr Opin Microbiol. 2: 148-152. Heyde, M. and Portalier, R. (1990). Acid Shock Proteins of Escherichia coli. FEMS Microbiol Lett. 57: 19-26. Hicks, S., Candy, D. C. and Phillips, A. D. (1996). Adhesion of Enteroaggregative Escherichia coli to Pediatric Intestinal Mucosa in Vitro. Infect Immun. 64: 4751-4760. Hill, C., O'Driscoll, B. and Booth, I. (1995). Acid Adaptation and Food Poisoning Microorganisms. Int J Food Microbiol 28: 245-254. Holt, J. G., Kieg, N. R., Sneath, P. H. A. and Staley, J. T. (1994). Bergry's Manual of Determinative Bacteriology. Williams and Wilkins Press. Maryland, USA. Iyer, R., Iverson, T. M., Accardi, A. and Miller, C. (2002). A Biological Role for Prokaryotic Clc Chloride Channels. Nature. 419: 715-718. Jordan, K. N., Oxford, L. and O'Byrne, C. P. (1999). Survival of Low-pH Stress by Escherichia coli O157:H7: Correlation Between Alterations in the Cell Envelope and Increased Acid Tolerance. Appl Environ Microbiol. 65: 3048-3055. Jungblut, P. and Thiede, B. 1997. Protein Identification from 2-DE Gels by MALDI Mass Spectrometry. Mass Spectrom. Rev. 16:145-162. Junkins, A. D. and Doyle, M. P. (1989). Comparison of Adherence Properties of Escherichia coli O157:H7 and 60 Megadalton Plasmid-Cured Derivative. Curr Microbiol. 19:21-27 Kim, H. H., Samadpour, M., Grimm, L., Clausen, C. R., Besser, T. E., Baylor, M., Kobayashi, J. M., Neill, M. A., Schoenknecht, F. D. and Tarr, P. I. (1994). Characteristics of Antibiotic-Resistant Escherichia coli O157:H7 in Washington State. J Infect Dis 170: 1606-1609. Konowalchuk, J., Speirs, J. I. and Stavric, S. (1977). Vero Response to a Cytotoxin of Escherichia coli. Infect Immunity 18: 775-779. Koutsoumanis, K. P. and Sofos, J. N. (2004). Comparative Acid Stress Response of Listeria Monocytogenes, Escherichia coli O157:H7 and Salmonella Typhimurium after Habituation at Different pH Conditions. Lett Appl Microbiol 38: 321-326. Lansbury, L. E. and Ludlam, L. (1997). Escherichia coli O157: Lessons from the Past 15 Yeats. J Infect. 34: 189-193. Leenanon, B. and Drake, M. A. (2001). Acid Stress, Starvation, and Cold Stress Affect Poststress Behavior of Escherichia coli O157:H7 and Nonpathogenic Escherichia coli. J Food Prot 64: 970-974. Leistner, L. (2000). Basic Aspects of Fod Preservation by Hurdle Technology. Int J Food Microbiol. 55: 181-186. Leyer, G. J., Wang, L. L. and Johnson, E. A. (1995). Acid adaptation of Escherichia coli O157:H7 Increases Survival in Acidic Foods. Appl Environ Microbiol. 61: 3752-3755. Lou, Y. and Yousef, A. E. (1997). Adaptation to Sublethal Environmental Stress Protects Listeria Monocytogenes Against Lethal Preservation Factors. Appl. Env Microbiol. 63: 1252-1255. Molenaar, D., Bosscher, J. S., Brink, B., Driessen, A. J. and Konings, W. N. J. (1993). Generation of a Proton Motive Force by Histidine Decarboxylation and Electrogenic Histidine/Histamine Antiport in Lactobacillus Buchneri. Bacteriol. 175: 2864-2870. Nataro, J. P. and Kaper, J. B. (1998). Diarrheagenic Escherichia coli. Clin Microbiol Rev. 11: 142-201. Nelson, D. L. and Cox, M. M. (2000). Lehninger Principles of Biochemistry. Worth Publishers. New York, USA. O’Farrell, P. H. (1975). High Resolution Two-dimensional Electrophoresis. J Biol Chem.. 250:4007-4021. Parkinson, J. S. and Kofoid, E. C. (1992). Communication Moduled in Bacterial Signaling Proteins. Ann Rev Genet. 26: 71-112. Price, S. B., Cheng, C. M., Kaspar, C. W., Wright, J. C., DeGraves, F. J., Penfound, T. A., Castanie-Cornet, M. P. and Foster, J. W. (2000). Role of RpoS in Acid Resistance and Fecal Shedding of Escherichia coli O157:H7. Appl Environ Microbiol 66: 632-637. Quinn, P. J., Carter, M. E., Markey, B. and Carter, G. R. (1994). Enterbacteriaceae. In: Clinical Veterinary Microbiology. London, UK. Ranford, J. C., Coates, A. R. M. and Henderson, B. (2000). Chaperonins Are Cell-Signalling Proteins: The Unfolding Biology of Molecular Chaperones. Exp Rev Mol Med. 15: 1-17. Rao, N. A., Ambili, M., Jala, V. R., Subramanya, H. S. and Savithri, H. S. (2003). Structure- Function Relationship in Serine Hydroxymethyltransferase. Biochimica et Biophysica Acta 1647: 24-29. Ratnam, S., March, S. B., Ahmed, R., Bezanson, G. S. and Kasatiya, S. (1988). Characterization of Escherichia coli Serotype O157:H7. J Clin Microbiol. 26: 2006-2012. Riley, L. W., Remis, R. S., Helgerson, S. D., McGee, H. B., Wells, J. G., Davis, B. R., Herbert, R. J., Olcott, E. S., Johnson, L. M., Hargrett, N. T., Blake, P. A. and Cohen, M. L. (1983). Hemorrhagic Colitis Associated with a Rare Escherichia coli Serotype. N Engl J Med. 308: 681-685. Sainz, T., Perez, J., Villaseca, J., Hernandez, U., Eslava, C., Mendoza, G. and Wacher, C. (2005). Survival to Different Acid Challenges and Outer Membrane Protein Profiles of Pathogenic Escherichia coli Strains Isolated from Pozol, a Mexican Typical Maize Fermented Food. Int J Food Microbiol 105: 357-367. Salmond, C. V., Kroll, R. G. and Booth, I. R. (1984). The Effect of Food Preservatives on pH Homeostasis in Escherichia coli. J Gen Microbiol 130: 2845-2850. Schardinger, F. (1892). The Occurrence of Fermentative Bacteria and Their Significance for the Sanitary Evaluation of Drinking Water. Wien Klin Wochenschr. 5:403-405, 421-432. Schimidt, H., Karch, H. and Benz, R. (1996). Pore-Forming Propertied of the Plasmid-Encoded Hemolysin of Enterohemorrhagic Escherichia coli O157:H7. Eur J Biochem. 241: 594-604. Seputiene, V., Motiejunas, D., Suziedelis, K., Tomenius, H., Normark, S., Melefors, O. and Suziedeliene, E. (2003). Molecular Characterization of the Acid-inducible asr Gene of Escherichia coli and Its Role in Acid Stress Response. J Bacteriol. 185: 2475-2484. Sharma, M., Adler, B. B., Harrison, M. D. and Beuchat, L. R. (2005). Thermal Tolerance of Acid-Adapted and Unadapted Salmonella, Escherichia coli O157:H7, and Listeria Monocytogenes in Cantaloupe Juice and Watermelon Juice. Lett Appl Microbiol 41: 448-453. Sugawara, E. and Nikaido, H. (1992). Pore-Forming Activity of ompA Protein of Escherichia coli. J Biol. Chem. 267: 2507-2511. Tsai, Y. W. and Ingham, S. C. (1997). Survival of Escherichia coli O157:H7 and Salmonella spp. in Acidic Condiments. J. Food Prot. 60: 751-755. Wasinger, V. C., Cordwell, S. J., Cerpa-Poljak, A., Yan, J. X., Gooley, A. A., Wilkins, M. R., Duncan, M. W., Harris, R., Williams, K. L. and Humphery-Smith, I. (1995). Progress with Gene-product Mapping of the Mollicutes: Mycoplasma Genitalium. Electrophoresis 16: 1090-1094. Wells, J. G., Shipman, L. D., Greene, K. D., Sowers, E. G., Green, J. H., Cameron, D. N., Downes, F. P., Martin, M. L., Griffin, P. M., Ostroff, S. M., Potter, M. E., Tauxe, R. V. and Wachsmuth, I. K. (1991). Isolation of Escherichia coli Serotype O157:H7 and Other Shiga-Like-Toxin-Producing E. coli from Dairy Cattle. J Clin Microbiol 29: 985-989. Wilmes-Riesenberg, M. R., Bearson, B., Foster, J. W. and Curtiss, R. (1996). Role of the Acid Tolerance Response in Virulence of Salmonella typhimurium. Infect. Immun. 64: 1085-1092. Wu, F. T., Tsai, T. Y., Hsu, C. F., Pan, T. M., Chen, H. Y. and Su, I. J. (2005) Isolation and Identification of Escherichia coli O157:H7 in a Taiwanese Patient with Bloody Diarrhea and Acute Renal Failure. J Formos Med Assoc. 104:206-209 Yates, J. R. 3rd (2000). Mass Spectrometry: From Genomics to Proteomics. Trends Genet. 16: 5-8. Yuk, H. G. and Marshall, D. L. (2005). Influence of Acetic, Citric, and Lactic Acids on Escherichia coli O157:H7 Membrane Lipid Composition, Verotoxin Secretion, and Acid Resistance in Simulated Gastric Fluid. J Food Prot 68: 673-679. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32123 | - |
dc.description.abstract | O157:H7 型大腸桿菌為重要之食品病原菌之ㄧ,與其他病原菌相比,此菌有較高之酸耐受性。根據前人研究發現,將此菌暴露於中等酸度環境一段時間,其會針對酸逆境進行調控,可能提高此菌對後續強酸 (如胃酸) 或是其他環境壓力之抵抗力,即產生酸耐性反應 (acid tolerance response, ATR)。本研究乃針對本土分離之 O157:H7 型大腸桿菌,觀察其於酸性環境下之生長情形,並對其產生酸耐性反應之蛋白質調控機制進行探討。
本研究使用民國 87 至 93 年間,自臺灣地區分離之 6 株 O157:H7 型大腸桿菌,包括 5 株家畜糞便分離株以及 1 株人體臨床檢體分離株,觀察其於酸性環境下之生長特性,結果顯示此 6 株本土分離之O157:H7 型大腸桿菌彼此間的酸耐受性並無顯著差異。繼而選擇其中人體臨床檢體分離株 TWC01,使用鹽酸及乳酸做為酸化劑,進行酸馴化 (acid adaptation) 處理,使其產生酸耐性反應,之後萃取馴化後菌體之總蛋白質,進行蛋白質體學分析,此外並觀察酸馴化對毒素分泌之影響。實驗結果發現,在毒素分泌方面,臨床分離株 TWC01 經酸馴化處理後有提高其 stx 毒素分泌的趨勢。分析二維電泳後之影像顯示,兩種酸馴化處理後之細胞,其蛋白質表現多為 up-regulation,且乳酸對細胞之影響大於鹽酸。以質譜分析表現量差異明顯的蛋白質發現,其主要為代謝相關之蛋白質,另有一部分與轉譯、蛋白質折疊,以及膜蛋白相關。由這些蛋白質於酸處理下表現量的變化得知,O157:H7 型大腸桿菌於酸處理下會產生許多大腸桿菌間共通之耐酸機制,同時代謝路徑傾向增加能量的生成以及 polypeptide 的合成,供細胞足夠之能量應付酸逆境所造成之生理變化,因而增加細胞對後續酸逆境之抗抵力,產生酸耐性反應;另一方面,O157:H7 型大腸桿菌細胞對環境中鹽酸 (無機酸) 與乳酸 (有機酸) 之感應,以及所引起之調控反應有所差異,顯示細胞需要較多之能量以應付環境中的乳酸。本研究有助於了解O157:H7 型大腸桿菌於酸逆境下之調控反應以及致病能力的影響,於食品安全方面,將可提供做為此菌防治上之參考。 | zh_TW |
dc.description.abstract | Escherichia coli O157:H7 is one of the important food-borne pathogens, and it has unusually higher resistance to acidic environment than other food-borne pathogens. Some researches revealed that acid adapted cells, by exposure to moderately acidic conditions, are more resistant to a subsequent strong acidic challenge or other stress. This study was conducted to understand the growth and the protein regulatory pathways of acid tolerance response (ATR) of local isolated E. coli O157:H7 induced by acidic environment.
Six local isolated E. coli O157:H7 were compared the growth under the acid stress and had no difference in acid tolerance. After that, chose the clinical strain TWC01 for further study. By using hydrochloric acid (HCl) or lactic acid as acidifier, E. coli O157:H7 TWC01 were acid adapted to induce ATR. The total protein of adapted cells were extracted for proteomic analysis. Furthermore, the effect of acid adaptation to stx secretion were examined. The result revealed that after acid adaptation, it tended to increase the stx secretion of E. coli O157:H7 TWC01. Image analysis of the gel indicated that a great part of proteins were up-regulation, and lactic acid had more effect on cell than HCl. Analyzed proteins by mass spectrometer, most of proteins were metabolism related protein, and others were related to translation, folding, and membrane proteins. According to protein expression variety under acid stress has found that the induction of many acid tolerance mechanism of E. coli. The metabolism pathway were toward increasing energy production and polypeptide synthesis, in order to provide enough energy and induce ATR. Furthermore, E. coli O157:H7 needed more energy to deal with lactic acid than HCl, revealed that organic and inorganic acid have different stress sensor and response system. This study provide the information of regulation and virulence of E. coli O157:H7 under acid stress, it may help control of this pathogen and improve food safety. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T03:32:45Z (GMT). No. of bitstreams: 1 ntu-95-R93b47102-1.pdf: 2406296 bytes, checksum: 456dfe2e58725ec34d738f62f784083f (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 摘 要 I
ABSTRACT II 目 錄 III 表 目 錄 V 圖 目 錄 VI 縮 寫 表 VIII 第一章 前言 1 第二章 文獻回顧 2 2.1 大腸桿菌 2 2.1.1 腸病原性大腸桿菌 4 2.1.2 腸毒性大腸桿菌 5 2.1.3 腸出血性大腸桿菌 5 2.1.4 腸侵襲性大腸桿菌 6 2.1.5 腸附著性大腸桿菌 6 2.1.6 擴黏型大腸桿菌 7 2.2 O157:H7 型大腸桿菌 9 2.2.1 生化特性 9 2.2.2 致病因子 12 2.2.3 感染與臨床症狀 13 2.2.4 傳染途徑 14 2.2.5 相關法規 15 2.3 微生物逆境反應 15 2.3.1 酸馴化反應 16 2.3.2 酸馴化機制 16 2.4 蛋白質體學 19 2.4.1 二維膠體電泳 19 2.4.2 蛋白質體學之應用 20 第三章 材料與方法 22 3.1 材料 22 3.1.1 菌株與培養條件 22 3.1.2 培養基 22 3.1.3 藥品與試劑 25 3.2 實驗儀器與設備 29 3.3 方法 30 3.3.1 本土分離 E. coli O157:H7 耐酸性比較 30 3.3.2 酸處理 30 3.3.3 Shiga-like toxin 定量 31 3.3.4 二維電泳分析 32 3.3.5 蛋白質質譜分析與身分鑑定 34 第四章 結果 39 4.1 本土分離之 O157:H7 型大腸桿菌酸耐性比較 39 4.2 臨床分離株 TWC01酸馴化處理後酸耐性之變化 42 4.3 酸馴化對 TWC01 細胞 STX 毒素分泌之影響 49 4.4 蛋白質體學 54 4.4.1 酸馴化過程對 TWC01 總蛋白質量表現變化 54 4.4.2 酸馴化對 TWC01 之蛋白質表現影響 56 4.4.3 質譜分析與資料庫比對結果 63 第五章 討論 70 5.1 本土分離之 O157:H7 型大腸桿菌酸耐性比較 70 5.2 酸馴化反應 70 5.3 兩種酸化劑對酸馴化之差異 71 5.4 酸馴化對毒素分泌之影響 73 5.5 蛋白質圖譜變化 73 5.5.1 代謝相關蛋白質 73 5.5.2遺傳訊息處理相關蛋白質 79 5.5.3環境訊息處理相關蛋白質 80 第六章 結論 82 第七章 參考文獻 83 | |
dc.language.iso | zh-TW | |
dc.title | 酸逆境下本土分離腸出血性大腸桿菌 O157:H7 TWC01之生理反應及蛋白質表現變化 | zh_TW |
dc.title | The physiological response and protein expression of local isolated Escherichia coli O157:H7 TWC01 under acid stress | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蘇遠志,黃健雄,游若?,林文源 | |
dc.subject.keyword | O157:H7 型大腸桿菌,酸耐性反應,二維電泳, | zh_TW |
dc.subject.keyword | Escherichia coli O157:H7,acid tolerance response,two-dimensional gel electrophoresis, | en |
dc.relation.page | 91 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-28 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-95-1.pdf 目前未授權公開取用 | 2.35 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。