人源性抗百草枯特异性单链抗体的筛选及初步鉴定
摘要
目的从大型人源性噬菌体抗体库中直接筛选出百草枯特异性单链抗体(scFv),并与百草枯人工抗原特异性结合性能进行鉴定。
方法以百草枯人工抗原(PQ-h-BSA)为靶抗原,采用固相化抗原免疫吸附筛选法,通过“吸附-洗脱-扩增”筛选富集过程,从大型人源性噬菌体抗体库中筛选出特异性百草枯单链抗体,通过ELISA法对其特异性结合性能进行初步鉴定。
结果经过5轮筛选,获得两株能与百草枯人工抗原特异性结合的阳性克隆。经ELISA法初步鉴定,筛选出的单链抗体与百草枯人工抗原反应强烈,而与其他抗原不反应或反应微弱。
结论利用噬菌体抗体技术可不经免疫制备人源性百草枯抗体,为临床治疗百草枯中毒研究提供了具有更广阔前景的人源性小分子抗体。
Objective To screen humanized anti-paraquat scFv from large human phage antibody library ,and its specific binding properties with paraquat artifical antigen were identified.
Methods In the process of "absorption - elution - amplification" , using solid-phase ELISA-based antigen screening method, that artificial antigen paraquat (PQ-h-BSA) as antigen,choosing by their specific binding properties of the initial identification in ELISA, to screen humanized anti-paraquat scFv from large human phage antibody library .
Results Two strains with paraquat artificial antigen-specific combination of positive clones were got in 5 rounds of screening. In initially identified by ELISA, anti-paraquat scFv of screeninig reacted strongly with paraquat artifical antigen , but no reaction or weak with other antigens.
Conclusion It is feasible to obtain humanized anti-paraquat scFv by phage display technique,which provides a more promising source of small molecule human antibodies for clinical treatment of paraquat poisoning study.
方法以百草枯人工抗原(PQ-h-BSA)为靶抗原,采用固相化抗原免疫吸附筛选法,通过“吸附-洗脱-扩增”筛选富集过程,从大型人源性噬菌体抗体库中筛选出特异性百草枯单链抗体,通过ELISA法对其特异性结合性能进行初步鉴定。
结果经过5轮筛选,获得两株能与百草枯人工抗原特异性结合的阳性克隆。经ELISA法初步鉴定,筛选出的单链抗体与百草枯人工抗原反应强烈,而与其他抗原不反应或反应微弱。
结论利用噬菌体抗体技术可不经免疫制备人源性百草枯抗体,为临床治疗百草枯中毒研究提供了具有更广阔前景的人源性小分子抗体。
Objective To screen humanized anti-paraquat scFv from large human phage antibody library ,and its specific binding properties with paraquat artifical antigen were identified.
Methods In the process of "absorption - elution - amplification" , using solid-phase ELISA-based antigen screening method, that artificial antigen paraquat (PQ-h-BSA) as antigen,choosing by their specific binding properties of the initial identification in ELISA, to screen humanized anti-paraquat scFv from large human phage antibody library .
Results Two strains with paraquat artificial antigen-specific combination of positive clones were got in 5 rounds of screening. In initially identified by ELISA, anti-paraquat scFv of screeninig reacted strongly with paraquat artifical antigen , but no reaction or weak with other antigens.
Conclusion It is feasible to obtain humanized anti-paraquat scFv by phage display technique,which provides a more promising source of small molecule human antibodies for clinical treatment of paraquat poisoning study.
引文
1. Dinis-Oliveira RJ.Sarmento A,Reis P,et a1.Acute paraquat poisoning:report of a survival case following intake of a potential lethal dose . Pediatr Emerg Care[J],2006,22:537-540.
2. Lin JL,Lin-Tan DT,Chen KH,et a1.Repeated pulse of methylprednisolone and cyelophosphamide with continuous dexamethasone therapy for patients with severe paraquat poisoning[J].Crit Care Med,2006,34:368-373.
3. Chen N,Bowles MR,Pond SM.Prevention of paraquat toxicity in suspensions of alveolar typeⅡcells by paraquat-specific antibodies[J].Hum Exp Toxicol,1994,13(8):551-557.
4. Aujame L,Geoffroy F,Sodoyer R. High affinity human antibodies by phage display[J]. Human Antibodies,1997,8:155-168.
5. McCafferty J, Griffiths A.D, Winter G, Chiswell DJ, Phage antibodies:filamentous phage displaying antibody variable domains[J].Nature 348(1990):552-554.
6. Smith GP, Filamentous fusion phage:novel expression vectors that display cloned antigens on the virion surface[J].Science 228(1985):1315-1317.
7. Wingter G, Synthetic human antibodies and a strategy for protein engineering[J].FEBS.Lett.430(1998):92-94.
8. Hassan M, Azzazy E, Highsmith WE, Phage display technology:clinical application and recent innovations[J]. Clin.Biochem.35(2002):425-445.
9.张国民,乔媛媛,陈宇萍,吕喆，安云庆等?哟笕萘渴删蹇固蹇馍秆∪嗽葱?抗角蛋白ScFv[J].首都医科大学学报,2007,28(1):71-74。
10.李春英,乔媛媛,王刚,陈宇萍,高天文,王琰等。从大容量噬菌体抗体库筛选人源性抗酪氨酸酶相关蛋白1抗体[J]。中国皮肤性病学杂志,2005,19(7):388-390.
11.夏敏。33例百草枯中毒的临床分析[J]。中国工业医学杂志,1995,8(5):278-279.
12. Rose MS, Smith LL.Evidence for energy dependent accumulation of paraquat into ratlung[J].Nature,1974,252:314-315.
13. Forman HJ,Aldrich TK,Posner MA,Fisher AB.Differential paraquat uptake and redox kinetics of rat grancclar pneumocytes and alveolar macrophages[J]. J.Pharmacol Exp Ther,1982,221:428-433.
14. Aldrich TK, Fisher AB,Candes E,Chance B. Evidence for lipid peroxidation by paraquat in the perfused lung[J]. Lab Invest,1983,60:473-485.
15.赵玉红,邱泽武。急性百草枯中毒及其治疗[J].药物不良反应杂志,2006,8(1):42-44。
16. Okonek S. hemoperfusion in toxicology: Basic considerations of its effectiveness[J]. Clin Toxicol,1981,18:1185~1198.
17. Proudfoot AT, Prescott LF, Jarvie DR. Haemodialysis for paraquat poisoning[J]. Hum Toxicol, 1987,6:69-74.
18. Devlin CM, Bowles MR, Gordon RB,et al. Production of a papraquat-specific murine single chain Fv fragment[J]. Biochem(Tokyo),1995,118(3):480-487.
19. Azzazy HM, Highsmith WE Jr. Phage display technology:Clinical applications and recent innovations [J].Clin B iochem,2002,35(6):425-445.
20.赵晓蓉,戴文涛,王敏,彭吉林,黄韬,王国斌,伍钢,沈关心等。表皮生长因子受体特异性单链抗体的筛选及初步鉴定[J]。广东药学院学报,2007,23(2):192-194。
21.杨婷婷,孙秀兰等百草枯人工抗原的合成及抗体的制备[J]。细胞与分子免疫学杂志,2008,24(9):924-927。
22. Brichta J, Vesela H, Franek M. Production of scFV recombinant fragments against 2,4-dichlorophenoxyacetic acid hapten using naive phage library[J]. Vet.Med. Czech,48,2003(9):237-247.
1. Hwang KY,Lee EY,Hong SY. Paraquat intoxication in korea[J]. Arch Environ Health,2002,57(2):162-166.
2.黄韶清,周玉淑,刘仁树。现代急性中毒诊断治疗学。1版。北京:人民军医出版社,2002:309-311.
3. Licker M,Schweizer A, Hohn L et al. Single lung transplantation for adult respiratory distress syndrome after paraquat poisoning[J].Thorax,1998,53(7):620.
4. Simth LL,Lewis CP,Wyatt I et al. The importance of epitheial uptake systems in lung toxicity[J].Environ Health Perspect,1990,85:25.
5. Yasaka T,Okudaira K,Fujito H et al. Further studies of lipid peroxidation in human paraquat poisoning[J]. Arch Intern Med,1986,146(4):681.
6. Fernandez Y,Anglade F,Mitjavila S. Paraquat and iron–dependent lipid peroxidation. NADPH versus NADPH-generating systems[J]. Biol Trace Elem Res,2000,74(3):191.
7. Van Remmen H,Qi W,Sabia M et al. Multiple deficiencies in antioxidant enzymes in mice result in a compound increase in sensitivity to oxidative stress[J] .Free radic biol Med,2004,36(12):1625
8. Tomita M,Okuyama T,Hidaka K. Changes in mRNAs of inducible intric oxide synthase and interleukin-1 beta in the liver,kidney and lung tissues of rats acutely exposed to paraquat[J].Leg Med(Tokyo),1999,1(3):127.
9.彭晓东,陈骥,梁创.急性百草枯中毒后MODS患者血清TNF-α和IL-10的变化及意义[J].中国实用医药,2008,3(14):9-11.
10. Gregory SW,Keith AG,Edgar CK.Lung function and airway inflammation in rats following exposure to combustion products of carbon-graphite/epoxy composite material:comparison to a rodent model of acute lung injury[J].Toxicology, 2003,183(1-3):175.
11. Dinis-Oliveira RJ,Sousa C,Remiao F,et al. Sodium salicylate preventsparaquat-induced apoptosis in the rat lung[J]. Free Radic Biol Med,2007,43(1):48-61.
12. Senator A,Rachidi W,Lehmann S,et al. Prion protects against DNA damage induced by paraquat in cultured cells[J]. Free Radical Biology & Medicine,2004,37(8): 1224-1230.
13. Satomi Y,Tsuchiya W,Mihara K, et al. Gene expression analysis of the lung following paraquat administration in rats using DNA microarray[J].Toxicol Sci,2004,29(2): 91-100.
14. Tomita M,Okuyama T,Katsuyama H, et al. Mouse model of paraquat-poisoned lungs and its gene expression profile[J].Toxicol(Ireland)2007,231(2-3):200-209.
15.曹钰,董玉龙,姚尧等.急性百草枯中毒所致急性肺损伤机制研究[J].中国呼吸与危重病监护杂志,2005,114(14):303.
16.佟飞,田英平,石汉文等.急性百草枯中毒大鼠肺组织中基质金属蛋白酶及其组织抑制剂表达变化的研究[J].中国急救医学,2006,26(1):41.
17. Walcarius A,Mouchotte R. Efficient in vitro paraquat removal via irreversible immobilization into zeolite particles. Arch Environ Contam Toxicol,2004,46(1): 135-140.
18. Kojima S,Miyazaki Y,Honda T, et al. Protective effects of dextran sulfate and polyvinyl sulfate against acute toxicity of paraquat in mice[J]. Toxicology,1991,69:93-99.
19.王永进,王泽慧.百草枯中毒治疗的研究进展[J].中国急救医学,2003,23:404-406.
20.鲁新.连续性血液灌流透析及激素冲击治疗百草枯中毒伴多器官功能衰竭[J].中国危重病急救医学,2008,20(7):448.
21. Koo JR,Kim JC. Failure of continuous venovenous Hemofiltration to prevent death in paraquat poisoning[J].Am J Kidney Dis,2002,39(1):55.
22. Toshiji, Katsumi T,Akihiko W, et al. Pretective effect of 3-methyl-1- phenyl- 2- pyrazolin-5-one,a free radical scavenger,on acute toxicity of paraquat in mice[J]. Toxicol Lett,2003,143(1):51-54.
23.崔燕,彭乱顺,杨洪霞。依达拉奉对百草枯中毒患者肺损伤的防治作用[J]。中国误诊学杂志,2008,8(1 5):3599-3600.
24.石汉文,胡军利,田英平等.异丙酚对大鼠百草枯中毒的干预研究[ J ].中华劳动卫生职业病杂志,2006, 24 ( 6) : 345 -348.
25.刘丽娜,赵金垣,陈莉,等.茶多酚对急性百草枯中毒大鼠治疗作用的实验研究[J].中华劳动卫生职业病杂志,2000,18:158-160.
26.佟飞.褪黑素对急性百草枯中毒大鼠肺组织核因子-κB及诱导型一氧化氮合酶的影响及治疗作用的研究[ J ] .中国现代医学杂志, 2006, 24 (16) : 3733– 3745.
27. Ortiz GG, Reiter RJ, Genotoxicity of paraquat micronuclei induced in bone marrow and peri pheral blood are inhibited by melatonin[ J ].Mutat Res, 2000, 464 (2) : 239– 245.
28. Karakashev P, Petrov L, Alexandrova A. Paraquat– induced lipid peroxidati on and injury in Ehrlich ascites tumor cells[ J ] .Neorplasmal, 2000, 47 (2) : 122– 124.
29. Bartoli GM, Muller A, Sies H,et al. Antioxidant effect of diethyldithiocarbamate on microsomal lipid peroxidation assessed by low-level chemiluminescence and alkaline production[J]. FEBS Lett,1998,164:371-374.
30.邵春风,常秀丽,吴强恩,班婷婷,姜楠,姚新民,周志俊。四氢吡啶二硫代氨基甲酸酯对百草枯致大鼠肺损伤的干预研究[J]。环境与职业医学,2007,24(2):157-160.
31. Venkaesan N. Pulmonary protective effects of curcumin against paraquat toxicity. Life Sci,2000,66(2):21-28.
32.苏建玲,李星海,佟飞,等.银杏叶提取物对急性百草枯中毒大鼠肺过氧化损伤的保护作用[J].中华劳动卫生职业病杂志.2003,21:226—227.
33. Ghazi-Khansari M, Nasiri G, Honarjoo M. Decreasing theoxidant stress from paraquat in isolated perfused rat lung using captopril and niacin [ J ] . Arch Toxicol, 2005, 79 ( 6) : 341-345.
34.马玉腾。氨溴索对百草枯中毒大鼠肺组织损伤及表面活性蛋白- A的影响[ J ]。中华劳动卫生职业病杂志, 2006,24 (6) : 348– 351.
35.夏国光,王丹琪,王艳等.糖皮质激素治疗急性呼吸窘迫综合征58例临床分析[J].中华全科医师杂志,2003,2(I):21.
36.粱东良,侯景玉,马海燕等.大剂量激素救治百草枯中毒的临床观察[J].中国急救医学,2000,20(11):690.
37. Inase N,Sawada M,Ohtani Y,et al.Cyelosporin A followed by the treatment of acute exacerbation of idiopathic pulmoⅫ-y fibrosis with eorticosteroid[J].Intern Med,2003,42(7):565—570.
38. Afzali S,Gholyaf M. The effectiveness of combined Treatment with Methylprednisolone and Cyclophosphamide in Oral paraquat poisoning[J]. Arch Iran Med,2008,11:387-391.
39.陈慧,石汉文,田英平.百草枯中毒致肺损伤的研究进展[J].临床荟萃,2006,21:146-148.
40. Vanderwal NA, Vanoirschot TF, Van Dijk A,et al. Mechanism of protection of alveolar typeⅡcells against paraquat-induced cytotoxicity by deferoxamine[J].Biochem Pharmacol,1990,39(11):1665-1671.
41. Chen N,Bowles MR,Pond SM. Prevention of paraquat toxicity in suspensions of alveolar typeⅡcells by paraquat-specific antibodies[J].Hum Exp Toxicol,1994, 13:551-557.
42.李晓岚,高景利,王爱田.早期血必净联合血液灌流治疗急性百草枯中毒的疗效观察[J].中国急救医学,2008,28:745-747.
43.谢东辉,章辉,郑兰荣.乌司他丁对百争枯中毒全身炎症反应综合征治疗作用及其机制的研究[J].实用医学杂志,2008,24:1339-1340.
44. Dinis-Oliveira RJ,Pontes H,Bastos ML,et al. An effective antidote for paraquat poisonings:The treatment with lysine acetylsalicylate[J].Toxicology,2009,255:187-193.
45.马玉腾,田英平,石汉文,等.大剂量氨溴索对百草枯中毒所致大鼠急性肺损伤的影响[J].中华劳动卫生职业病杂志,2007,25:523-526.
46. Licker M,Schweizer A,Hohn L,el a1.Single lung transplantation for adult respiratory distress syndrome after paraquat poisoning[J]. Thorax,1998,53:620-621
47. Gear AJ,Ahrenholz DH,Solem LD.Paraquat poisoning in a burn patient.J Burn Care Rehabil[J].200I,22:347-351.
48. Hagiwara S,Iwasaka H,Matsumoto S,et a1.An antisense oligonueleotide to HSP47 inhibits paraquat—induced pulmonary fibrosis in rats[J].Toxicology,2007,236: l99-207.
2. Lin JL,Lin-Tan DT,Chen KH,et a1.Repeated pulse of methylprednisolone and cyelophosphamide with continuous dexamethasone therapy for patients with severe paraquat poisoning[J].Crit Care Med,2006,34:368-373.
3. Chen N,Bowles MR,Pond SM.Prevention of paraquat toxicity in suspensions of alveolar typeⅡcells by paraquat-specific antibodies[J].Hum Exp Toxicol,1994,13(8):551-557.
4. Aujame L,Geoffroy F,Sodoyer R. High affinity human antibodies by phage display[J]. Human Antibodies,1997,8:155-168.
5. McCafferty J, Griffiths A.D, Winter G, Chiswell DJ, Phage antibodies:filamentous phage displaying antibody variable domains[J].Nature 348(1990):552-554.
6. Smith GP, Filamentous fusion phage:novel expression vectors that display cloned antigens on the virion surface[J].Science 228(1985):1315-1317.
7. Wingter G, Synthetic human antibodies and a strategy for protein engineering[J].FEBS.Lett.430(1998):92-94.
8. Hassan M, Azzazy E, Highsmith WE, Phage display technology:clinical application and recent innovations[J]. Clin.Biochem.35(2002):425-445.
9.张国民,乔媛媛,陈宇萍,吕喆，安云庆等?哟笕萘渴删蹇固蹇馍秆∪嗽葱?抗角蛋白ScFv[J].首都医科大学学报,2007,28(1):71-74。
10.李春英,乔媛媛,王刚,陈宇萍,高天文,王琰等。从大容量噬菌体抗体库筛选人源性抗酪氨酸酶相关蛋白1抗体[J]。中国皮肤性病学杂志,2005,19(7):388-390.
11.夏敏。33例百草枯中毒的临床分析[J]。中国工业医学杂志,1995,8(5):278-279.
12. Rose MS, Smith LL.Evidence for energy dependent accumulation of paraquat into ratlung[J].Nature,1974,252:314-315.
13. Forman HJ,Aldrich TK,Posner MA,Fisher AB.Differential paraquat uptake and redox kinetics of rat grancclar pneumocytes and alveolar macrophages[J]. J.Pharmacol Exp Ther,1982,221:428-433.
14. Aldrich TK, Fisher AB,Candes E,Chance B. Evidence for lipid peroxidation by paraquat in the perfused lung[J]. Lab Invest,1983,60:473-485.
15.赵玉红,邱泽武。急性百草枯中毒及其治疗[J].药物不良反应杂志,2006,8(1):42-44。
16. Okonek S. hemoperfusion in toxicology: Basic considerations of its effectiveness[J]. Clin Toxicol,1981,18:1185~1198.
17. Proudfoot AT, Prescott LF, Jarvie DR. Haemodialysis for paraquat poisoning[J]. Hum Toxicol, 1987,6:69-74.
18. Devlin CM, Bowles MR, Gordon RB,et al. Production of a papraquat-specific murine single chain Fv fragment[J]. Biochem(Tokyo),1995,118(3):480-487.
19. Azzazy HM, Highsmith WE Jr. Phage display technology:Clinical applications and recent innovations [J].Clin B iochem,2002,35(6):425-445.
20.赵晓蓉,戴文涛,王敏,彭吉林,黄韬,王国斌,伍钢,沈关心等。表皮生长因子受体特异性单链抗体的筛选及初步鉴定[J]。广东药学院学报,2007,23(2):192-194。
21.杨婷婷,孙秀兰等百草枯人工抗原的合成及抗体的制备[J]。细胞与分子免疫学杂志,2008,24(9):924-927。
22. Brichta J, Vesela H, Franek M. Production of scFV recombinant fragments against 2,4-dichlorophenoxyacetic acid hapten using naive phage library[J]. Vet.Med. Czech,48,2003(9):237-247.
1. Hwang KY,Lee EY,Hong SY. Paraquat intoxication in korea[J]. Arch Environ Health,2002,57(2):162-166.
2.黄韶清,周玉淑,刘仁树。现代急性中毒诊断治疗学。1版。北京:人民军医出版社,2002:309-311.
3. Licker M,Schweizer A, Hohn L et al. Single lung transplantation for adult respiratory distress syndrome after paraquat poisoning[J].Thorax,1998,53(7):620.
4. Simth LL,Lewis CP,Wyatt I et al. The importance of epitheial uptake systems in lung toxicity[J].Environ Health Perspect,1990,85:25.
5. Yasaka T,Okudaira K,Fujito H et al. Further studies of lipid peroxidation in human paraquat poisoning[J]. Arch Intern Med,1986,146(4):681.
6. Fernandez Y,Anglade F,Mitjavila S. Paraquat and iron–dependent lipid peroxidation. NADPH versus NADPH-generating systems[J]. Biol Trace Elem Res,2000,74(3):191.
7. Van Remmen H,Qi W,Sabia M et al. Multiple deficiencies in antioxidant enzymes in mice result in a compound increase in sensitivity to oxidative stress[J] .Free radic biol Med,2004,36(12):1625
8. Tomita M,Okuyama T,Hidaka K. Changes in mRNAs of inducible intric oxide synthase and interleukin-1 beta in the liver,kidney and lung tissues of rats acutely exposed to paraquat[J].Leg Med(Tokyo),1999,1(3):127.
9.彭晓东,陈骥,梁创.急性百草枯中毒后MODS患者血清TNF-α和IL-10的变化及意义[J].中国实用医药,2008,3(14):9-11.
10. Gregory SW,Keith AG,Edgar CK.Lung function and airway inflammation in rats following exposure to combustion products of carbon-graphite/epoxy composite material:comparison to a rodent model of acute lung injury[J].Toxicology, 2003,183(1-3):175.
11. Dinis-Oliveira RJ,Sousa C,Remiao F,et al. Sodium salicylate preventsparaquat-induced apoptosis in the rat lung[J]. Free Radic Biol Med,2007,43(1):48-61.
12. Senator A,Rachidi W,Lehmann S,et al. Prion protects against DNA damage induced by paraquat in cultured cells[J]. Free Radical Biology & Medicine,2004,37(8): 1224-1230.
13. Satomi Y,Tsuchiya W,Mihara K, et al. Gene expression analysis of the lung following paraquat administration in rats using DNA microarray[J].Toxicol Sci,2004,29(2): 91-100.
14. Tomita M,Okuyama T,Katsuyama H, et al. Mouse model of paraquat-poisoned lungs and its gene expression profile[J].Toxicol(Ireland)2007,231(2-3):200-209.
15.曹钰,董玉龙,姚尧等.急性百草枯中毒所致急性肺损伤机制研究[J].中国呼吸与危重病监护杂志,2005,114(14):303.
16.佟飞,田英平,石汉文等.急性百草枯中毒大鼠肺组织中基质金属蛋白酶及其组织抑制剂表达变化的研究[J].中国急救医学,2006,26(1):41.
17. Walcarius A,Mouchotte R. Efficient in vitro paraquat removal via irreversible immobilization into zeolite particles. Arch Environ Contam Toxicol,2004,46(1): 135-140.
18. Kojima S,Miyazaki Y,Honda T, et al. Protective effects of dextran sulfate and polyvinyl sulfate against acute toxicity of paraquat in mice[J]. Toxicology,1991,69:93-99.
19.王永进,王泽慧.百草枯中毒治疗的研究进展[J].中国急救医学,2003,23:404-406.
20.鲁新.连续性血液灌流透析及激素冲击治疗百草枯中毒伴多器官功能衰竭[J].中国危重病急救医学,2008,20(7):448.
21. Koo JR,Kim JC. Failure of continuous venovenous Hemofiltration to prevent death in paraquat poisoning[J].Am J Kidney Dis,2002,39(1):55.
22. Toshiji, Katsumi T,Akihiko W, et al. Pretective effect of 3-methyl-1- phenyl- 2- pyrazolin-5-one,a free radical scavenger,on acute toxicity of paraquat in mice[J]. Toxicol Lett,2003,143(1):51-54.
23.崔燕,彭乱顺,杨洪霞。依达拉奉对百草枯中毒患者肺损伤的防治作用[J]。中国误诊学杂志,2008,8(1 5):3599-3600.
24.石汉文,胡军利,田英平等.异丙酚对大鼠百草枯中毒的干预研究[ J ].中华劳动卫生职业病杂志,2006, 24 ( 6) : 345 -348.
25.刘丽娜,赵金垣,陈莉,等.茶多酚对急性百草枯中毒大鼠治疗作用的实验研究[J].中华劳动卫生职业病杂志,2000,18:158-160.
26.佟飞.褪黑素对急性百草枯中毒大鼠肺组织核因子-κB及诱导型一氧化氮合酶的影响及治疗作用的研究[ J ] .中国现代医学杂志, 2006, 24 (16) : 3733– 3745.
27. Ortiz GG, Reiter RJ, Genotoxicity of paraquat micronuclei induced in bone marrow and peri pheral blood are inhibited by melatonin[ J ].Mutat Res, 2000, 464 (2) : 239– 245.
28. Karakashev P, Petrov L, Alexandrova A. Paraquat– induced lipid peroxidati on and injury in Ehrlich ascites tumor cells[ J ] .Neorplasmal, 2000, 47 (2) : 122– 124.
29. Bartoli GM, Muller A, Sies H,et al. Antioxidant effect of diethyldithiocarbamate on microsomal lipid peroxidation assessed by low-level chemiluminescence and alkaline production[J]. FEBS Lett,1998,164:371-374.
30.邵春风,常秀丽,吴强恩,班婷婷,姜楠,姚新民,周志俊。四氢吡啶二硫代氨基甲酸酯对百草枯致大鼠肺损伤的干预研究[J]。环境与职业医学,2007,24(2):157-160.
31. Venkaesan N. Pulmonary protective effects of curcumin against paraquat toxicity. Life Sci,2000,66(2):21-28.
32.苏建玲,李星海,佟飞,等.银杏叶提取物对急性百草枯中毒大鼠肺过氧化损伤的保护作用[J].中华劳动卫生职业病杂志.2003,21:226—227.
33. Ghazi-Khansari M, Nasiri G, Honarjoo M. Decreasing theoxidant stress from paraquat in isolated perfused rat lung using captopril and niacin [ J ] . Arch Toxicol, 2005, 79 ( 6) : 341-345.
34.马玉腾。氨溴索对百草枯中毒大鼠肺组织损伤及表面活性蛋白- A的影响[ J ]。中华劳动卫生职业病杂志, 2006,24 (6) : 348– 351.
35.夏国光,王丹琪,王艳等.糖皮质激素治疗急性呼吸窘迫综合征58例临床分析[J].中华全科医师杂志,2003,2(I):21.
36.粱东良,侯景玉,马海燕等.大剂量激素救治百草枯中毒的临床观察[J].中国急救医学,2000,20(11):690.
37. Inase N,Sawada M,Ohtani Y,et al.Cyelosporin A followed by the treatment of acute exacerbation of idiopathic pulmoⅫ-y fibrosis with eorticosteroid[J].Intern Med,2003,42(7):565—570.
38. Afzali S,Gholyaf M. The effectiveness of combined Treatment with Methylprednisolone and Cyclophosphamide in Oral paraquat poisoning[J]. Arch Iran Med,2008,11:387-391.
39.陈慧,石汉文,田英平.百草枯中毒致肺损伤的研究进展[J].临床荟萃,2006,21:146-148.
40. Vanderwal NA, Vanoirschot TF, Van Dijk A,et al. Mechanism of protection of alveolar typeⅡcells against paraquat-induced cytotoxicity by deferoxamine[J].Biochem Pharmacol,1990,39(11):1665-1671.
41. Chen N,Bowles MR,Pond SM. Prevention of paraquat toxicity in suspensions of alveolar typeⅡcells by paraquat-specific antibodies[J].Hum Exp Toxicol,1994, 13:551-557.
42.李晓岚,高景利,王爱田.早期血必净联合血液灌流治疗急性百草枯中毒的疗效观察[J].中国急救医学,2008,28:745-747.
43.谢东辉,章辉,郑兰荣.乌司他丁对百争枯中毒全身炎症反应综合征治疗作用及其机制的研究[J].实用医学杂志,2008,24:1339-1340.
44. Dinis-Oliveira RJ,Pontes H,Bastos ML,et al. An effective antidote for paraquat poisonings:The treatment with lysine acetylsalicylate[J].Toxicology,2009,255:187-193.
45.马玉腾,田英平,石汉文,等.大剂量氨溴索对百草枯中毒所致大鼠急性肺损伤的影响[J].中华劳动卫生职业病杂志,2007,25:523-526.
46. Licker M,Schweizer A,Hohn L,el a1.Single lung transplantation for adult respiratory distress syndrome after paraquat poisoning[J]. Thorax,1998,53:620-621
47. Gear AJ,Ahrenholz DH,Solem LD.Paraquat poisoning in a burn patient.J Burn Care Rehabil[J].200I,22:347-351.
48. Hagiwara S,Iwasaka H,Matsumoto S,et a1.An antisense oligonueleotide to HSP47 inhibits paraquat—induced pulmonary fibrosis in rats[J].Toxicology,2007,236: l99-207.