鸭疫里默氏杆菌（RA）人工感染雏鹅的病理学变化和免疫组化检测RA抗原方法的建立及其RA抗原在鹅体组织器官分布规律的研究
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摘要
本研究以实验病理学方法对人工感染鹅源1型鸭疫里默氏杆菌(RA)的21日龄的雏鹅进行了系统病理组织学的研究，建立了RA感染雏鹅的病理模型。感染30小时后，急性死鹅1只，其他鹅出现典型的临床症状；感染46到72小后，半数鹅死亡，达到死亡高峰；感染9天后，20％鹅耐过。大体解剖病变为全身各组织器官的浆膜面广泛的纤维素性渗出；肝脏、胸腺肿大；脾脏、法氏囊感染前期肿大，感染后期萎缩；肠道出血，盲肠前期出血，后期畸变；皮下充血出血，并有胶冻样浸润，脑充血。病理组织学变化为纤维素性坏死性心肌炎、肝周炎、脑膜炎；急性肾小管坏死性肾炎、胰腺炎、肠炎；出血性胸腺炎、肺炎、脾炎等；盲肠扁桃体、脾脏、胸腺淋巴滤泡中淋巴细胞减少；除胰腺外，所有检测的组织中都有大量嗜中性白细胞出现。
本研究建立了间接免疫组化(Indirect Immunoperoxidase Staining,IIS)检测RA抗原的方法，该法的最适条件为：以PLP为固定液，以APES为粘片剂，以0.3％的甲醇-H_2O_2在湿盒中室温下孵育20分钟消除组织中的过氧化物酶及假过氧化物酶活性，兔抗RAIgG与羊抗兔—HRP的稀释比例均为1：25，以4℃过夜作兔抗RAIgG的孵育条件，以10％白蛋白0.05％Tween-20pH7.4的PBS湿盒内室温孵育60分钟处理组织中非特异性染色因素；以TBST为洗涤液，空气振荡器70转/分钟摇洗两次，每次10分钟洗涤切片；以0.5％甲苯胺蓝作为复染液。该方法用于检测感染雏鹅组织器官中的RA抗原具有特异、敏感、直观的优点。
本研究应用建立的间接免疫组化法对RA人工感染21日龄雏鹅后RA抗原在雏鹅各组织器官的分布规律检测结果表明，鹅人工感染RA24小时后，心脏、肝脏、脾脏、肾脏、回肠和直肠RA抗原呈强阳性分布，其次在肺脏、十二指肠、空肠和盲肠RA抗原呈弱阳性分布，胰腺、胸腺、法氏囊和大脑为阴性。在感染46小时后，所有检测器官中都呈阳性，大部分RA抗原阳性组织(肾脏、肺脏、胸腺、十二指肠、回肠、大脑)中都出现嗜甲苯胺蓝物质。在感染58小时后，各个器官RA抗原呈强阳性分布。感染第5天后肝细胞和肺房上皮细胞RA抗原分布呈阳性，感染9天后，肝、肾、胸腺的阳性强度没有下降，其他器官的阳性强度都下降。整个感染过程中，空肠和胰腺RA抗原阳性强度分布较低，大脑RA抗原阳性分布最低。各组织器官中RA抗原阳性分布呈现从间质到实质、从血管及血管周围到组织细胞规律分布的特点。RA感染对鹅的实质器官、肠道、免疫器官都有严重的损伤；RA抗原的分布与RA感染途径及其机体的抵抗力有关；RA对雏鹅机体组织器官的损伤程度与RA抗原的分布阳性强度及其嗜中性白细胞的浸润呈正相关。
Geese were artificially infected with Remerella anatipestifer serotype 1 at 21 days old to study the morphological pathology and to establish indirect immunohistochemistry methods to detect RA antigen and RA antigen distribution in infected geese. 30 hours post infected(PI) ,one goose was died and most of the others demonstrated clinical signs which were similar to that of duckings infected with RA; 46 to 72 hours PI, 50% of the goose showed highest mortality; 22 days PI20% of the geese survived. The gross pathological changes were as follows: fibrins on the seromembrane of all the organs, especially on the heart, liver, air sac and meningitis; swelling of liver and thymus ,swelling of spleen and bursal of Fabricius in early period and atrophy in the later stage, hyperemia and hemorrhage of the intestine, aberration of cecum in the later stage. Many parenchyma organs of infected geese showed fibrins and necrosis inflammation. Intestine showed hemorrhage inflammation. Few lymphocyte was found in spleen, thymu
s, bursa of fabricius and cecal tosil. Neutrophils infiltration could be seen in most of the organs.
The best conditions for indirect immunochemistry were as follows: Fixing specimens with PLP, processing slides with APES, eliminating the activity of catalase and its similar enzyme with 0.3% formaldehyde-H2O2, blocking the tissue section with 10% albumin-tween-20 in humidity, restaining with toluidine blue. It was found that this method showed great specificity and sensitivity.
The result of indirect immunochemistry demonstrated that: 24 hours PI, heart, liver, spleen, kidney, ileum, rectum, lung, duodenum and cecum were positive; 46 hours PI, all the organs showed positive results and some of the organs showed heterophile substances which could be stained by toluidine blue; 58 hours PI, the positive results were most obvious in all organs; 5 days PI, alveloar endothelial cells and hepatocytes showed positive in immunochemistry detection; 9 days PI, liver, kidney and thymus still showed strong results in immunohistochemistry detection while that of other organs began to decline; jejunum and pancreas showed less obvious results compared with other organs listed above in Immunohistochemistry detection
and brain was the least obvious.
In summary, RA infection in geese could cause severe damages to parenchyma organs, intestines and the immune organs; pathological lesions were correlated with the distribution of RA antigen and neutrophils.
本研究建立了间接免疫组化(Indirect Immunoperoxidase Staining,IIS)检测RA抗原的方法，该法的最适条件为：以PLP为固定液，以APES为粘片剂，以0.3％的甲醇-H_2O_2在湿盒中室温下孵育20分钟消除组织中的过氧化物酶及假过氧化物酶活性，兔抗RAIgG与羊抗兔—HRP的稀释比例均为1：25，以4℃过夜作兔抗RAIgG的孵育条件，以10％白蛋白0.05％Tween-20pH7.4的PBS湿盒内室温孵育60分钟处理组织中非特异性染色因素；以TBST为洗涤液，空气振荡器70转/分钟摇洗两次，每次10分钟洗涤切片；以0.5％甲苯胺蓝作为复染液。该方法用于检测感染雏鹅组织器官中的RA抗原具有特异、敏感、直观的优点。
本研究应用建立的间接免疫组化法对RA人工感染21日龄雏鹅后RA抗原在雏鹅各组织器官的分布规律检测结果表明，鹅人工感染RA24小时后，心脏、肝脏、脾脏、肾脏、回肠和直肠RA抗原呈强阳性分布，其次在肺脏、十二指肠、空肠和盲肠RA抗原呈弱阳性分布，胰腺、胸腺、法氏囊和大脑为阴性。在感染46小时后，所有检测器官中都呈阳性，大部分RA抗原阳性组织(肾脏、肺脏、胸腺、十二指肠、回肠、大脑)中都出现嗜甲苯胺蓝物质。在感染58小时后，各个器官RA抗原呈强阳性分布。感染第5天后肝细胞和肺房上皮细胞RA抗原分布呈阳性，感染9天后，肝、肾、胸腺的阳性强度没有下降，其他器官的阳性强度都下降。整个感染过程中，空肠和胰腺RA抗原阳性强度分布较低，大脑RA抗原阳性分布最低。各组织器官中RA抗原阳性分布呈现从间质到实质、从血管及血管周围到组织细胞规律分布的特点。RA感染对鹅的实质器官、肠道、免疫器官都有严重的损伤；RA抗原的分布与RA感染途径及其机体的抵抗力有关；RA对雏鹅机体组织器官的损伤程度与RA抗原的分布阳性强度及其嗜中性白细胞的浸润呈正相关。
Geese were artificially infected with Remerella anatipestifer serotype 1 at 21 days old to study the morphological pathology and to establish indirect immunohistochemistry methods to detect RA antigen and RA antigen distribution in infected geese. 30 hours post infected(PI) ,one goose was died and most of the others demonstrated clinical signs which were similar to that of duckings infected with RA; 46 to 72 hours PI, 50% of the goose showed highest mortality; 22 days PI20% of the geese survived. The gross pathological changes were as follows: fibrins on the seromembrane of all the organs, especially on the heart, liver, air sac and meningitis; swelling of liver and thymus ,swelling of spleen and bursal of Fabricius in early period and atrophy in the later stage, hyperemia and hemorrhage of the intestine, aberration of cecum in the later stage. Many parenchyma organs of infected geese showed fibrins and necrosis inflammation. Intestine showed hemorrhage inflammation. Few lymphocyte was found in spleen, thymu
s, bursa of fabricius and cecal tosil. Neutrophils infiltration could be seen in most of the organs.
The best conditions for indirect immunochemistry were as follows: Fixing specimens with PLP, processing slides with APES, eliminating the activity of catalase and its similar enzyme with 0.3% formaldehyde-H2O2, blocking the tissue section with 10% albumin-tween-20 in humidity, restaining with toluidine blue. It was found that this method showed great specificity and sensitivity.
The result of indirect immunochemistry demonstrated that: 24 hours PI, heart, liver, spleen, kidney, ileum, rectum, lung, duodenum and cecum were positive; 46 hours PI, all the organs showed positive results and some of the organs showed heterophile substances which could be stained by toluidine blue; 58 hours PI, the positive results were most obvious in all organs; 5 days PI, alveloar endothelial cells and hepatocytes showed positive in immunochemistry detection; 9 days PI, liver, kidney and thymus still showed strong results in immunohistochemistry detection while that of other organs began to decline; jejunum and pancreas showed less obvious results compared with other organs listed above in Immunohistochemistry detection
and brain was the least obvious.
In summary, RA infection in geese could cause severe damages to parenchyma organs, intestines and the immune organs; pathological lesions were correlated with the distribution of RA antigen and neutrophils.
引文
1. Sandhu, T,S.,and E.G. Harry. Serotypes of Pasteurella anatipestifer isolated from comm. Erical White Pekin ducks in the United States[J].Avian Dis, 1981,25;497-502.
2. Sandhu, T.S, and M. Leis Serotypes of Pasteurella anatipestifer isolates from poultry in different countries[il. Avian Pathol, 1991,20:233-239.
3．郭玉璞，陈德威，范国雄等．北京小鸭传染性浆膜炎的调查研究[J]．畜牧兽医学报，1982，12(2)：107-112．
4．刘凌云、郑光美主编《普通动物学》(第三版)，高等教育出版社，1997。
5．B．W 卡尔尼克主．高福，苏敬良，主译．禽病学[M]，(第十版)．中国农业出版社，1999，195—20。
6. Sandhu, T.S. immunogenicity and safety of a live pasteurella anatipestifer vaccine in White Pekin ducklings; Laboratory and field trials[J] Avian Pathol, 1991,20:423-432。
7. {Helfer, D.H.Pasteurella anatipestifer infection in Turkeys[J].Avian Dis, 1973,21(4):712-715.
8. Pierce, R.l. and M.W. Vorhies. Pasteurella anatipestifer infection in geese[J].Avian Dis, 1973,17:868-870.
9. Smith, J.M,D.D. Frame, G. Cooper et. al Pasteurella anatipestifer infetion in comm. ercial Meat-type Tokrkeys in California[J].Avian Diseases, 1987,31(5):913-917.
10. Frommer, A.,R. Bock, A. Inbar, and S. Zemer. Muscovy ducks as a source of Pasteurella anatipestifer infection in turkey flocks [J]. Avian Pathol, 1990,19:161-163.
11．蔡文琴，王伯沄主编．实用免疫细胞化学与核酸分子杂交技术．四川科学技术出版社，1994．72-97．4．
12．纪小龙，王淑琴，李红芬．免疫组化在临床诊断中的作用．Chin J Lab Digan, April 1998, Vol 2, No.2．
13. Bisgaard, M Antigenic studies on pasteurella anatipestifer species in certae sedis, Using slide and tube agglutingation[J].Avain pathology, 1982,11:341-350.
14．郭玉璞，甘孟候，张中直．应用荧光抗体法诊断小鸭传染性浆膜炎的实验[J]．中国兽医杂志，1982，8(7)：15-17．
15．苏敬良，黄瑜，吕艳丽．间接荧光抗体技术检测鸭疫里氏杆菌[J]．中国兽医杂志，1999，25(2)：12-13．
16．李桂杰，朱瑞良，郭宝聚．细菌内毒素的研究进展．山东农业大学学报，1999，29
30(2)．
17．许国根，陈突，张万光，等．对于全身炎症反应综合征患者致炎因子与抗炎因子的变化及相关性研究．Chin J Crit Care med june 2002．22(6)．
18．内毒素的产生，(见网页：毒与健康—毒的揭示 2000 见沈阳飞龙制药有限公司，E-mail：webmaster@medr.com)
19．陈君涛，毒素对内皮细胞的损伤机制，第三军医大学西南医院烧伤研究所，Foreign Medical Sciences Section of PathopHysiology．
20．Bevilacqua MP, Paber JS, Wheeler ME, et al. Interleukin 1 acts on cultures haman vascular endothelium to increase the adhesiOn of polymorepHnuclear leukocytes, monocytes and related leukocyte cell lines. J Clin Invest 1985, 76: 2003.
21．陈敏英，管向东．多器官功能障碍综合征的病理生理．中国实用妇科与产科杂志，2002，18(6)．
22．陈建．内毒素几何效应细胞的分子机制，第三军医大学附属新安医院烧伤研究所，重庆，第三军医大学学报，1999，21(11)．
23．陈国强，杨凌，杨永宗．内毒素在动脉粥样硬化形成中的可能作用．衡阳医学院心血管病研究所，生理科学进展，1991，22(4)．
24．陈敏英．多器官功能障碍综合征的病理生理．中国实用妇科与产科杂志，2002，18(6)．
25．许国根，王戈，缪群，等．全身炎症反应综合征患者抗炎因子对T淋巴细胞．凋亡的影响，中华急诊医学杂志，2002，8，11(4)．
26．齐文广，任爱民，张淑文，等．血清促炎、抗炎因子在急性感染患者中的动态变化及中药干预的影响．中国中西医结合杂志，2000，20(11)．
27．夏时海，赵晓晏，郭萍 急性胰腺炎促炎因子和抗炎因子变化的动态观察 第三军医大学附属新桥医院消化内科、重庆侧37第三军医大学学报 2001 23(4))。
28．罗昆仑．嗜中性白细胞介导组织损伤的机理．第三军医大学西南医院肝胆外科，国外医学生理、病理科学与临床分册，1999，19(1)．
29．周向东，陈意生，史景泉，等．内毒素条件下参与嗜中性白细胞—血管内皮细胞粘附过程的分子基础，第三军医大学基础部病理学教研室，第三军医大学学报，1996，8(2)．
30．李春盛，桂培春，何新华，等．NO和iNOS在内毒素诱导的大鼠急性肺损伤的作用极大黄对其影响．中国急救医学，2000，20(2)．
31．汪盛贤，马怖仁，于忠和，等．内毒素性肺损伤大鼠肺表面活性物质的变化．成都军区程度总医院，解放军医学杂志，1997，22(2)．
32．王伟阳．循环内毒素促进肠道内毒素的转运及选择性肠道消毒的影响．病理生理学教研室，北京医科大学学报1993 25(3)．
33．Deitch EA. et al Eadotoxin promotes the transtocation of bacteria from the
gut. Arch Surg 1987: 122:185.
34. O/Dwyer ST, et al.A single dose of endotoxin inzrcases intestinal permeability in healthy humans. Arch Surg 1988;123:1459.
35. Noman JG, Fink G; Frane M et al, Guffay J, Garter G, Dlviyon B, Sexton C, Glaccum M Aruive incerleucin-1 receptor requied for maximal pregression of accte pancrearitis. Am surge,1996;223:163-169.
36. Kimural k, Shimosequece I, Ahe R, Masamine A, Sarah A, Takasa A, Koizumi M, Toyote T. Low dose of lipopolysecchoride upreguiore zccrar cell apoptosis in cenuleir, pancneatitis panxreas, 1998;17:120-126.
37．彭小宁．内毒素诱导淋巴细胞胞凋亡．附属湘雅医院传染科，湖南医科大学学报 2000，25(6)．
38. Negate S.Apoptosis by death faxtor[J]. Cell,1997,28.
39. Barke RA, Roy S .Chapin RB. et al. The induction of accelerated thymic programmed cell death during polymicrobial sepsis: control by corticosceroids but tumor dcriosis factor. Shock. 1995.3:259-267.
40. Parksm A, Bullcy GB, Granger DN, et al.Role of oxygen-free redical in shock, ischemia, and organ preservation. Surgery, 1983,94(3):428-430.
41. PHung C D ,Ezineme T A, Turrens J F, Hydrogen peroxide metabolism in muscle mitochondria, Arch Biochem BiopHys, 1994,315(2),479-482.
42．陆松敏，刘建仓，李萍．毒素休克大鼠肝线粒体质子跨膜转运的改变．第三军医大学野战外科研究所，生物化学与生物物理进展，1999；26(1)．
43．郭德玉，陈意生．实验性内毒素介导的大鼠肾脏病理变化．第三军医大学基础部病理学教研室，第三军医大学学报，1995；17(6)．
44．雄正文，王泊云．影响免疫组织抗原性的因素分析与对策诊断病理学杂志．1997；9：171．
45．熊正文，胡海霞，李春光．盐酸水解暴露抗原促进免疫组织化学反应的研究．中国组织化学与细胞化学杂志，1996；5：71．
46．王伟阳，内毒素血症大鼠肠道内毒素移位的研究，北京医科大学生理学系生理科学进展 1995；26(1)．
47．内源性内毒素移位在失血性休克中的作用及其分子机理研究，第三军医大学外科学 1995；12月．
2. Sandhu, T.S, and M. Leis Serotypes of Pasteurella anatipestifer isolates from poultry in different countries[il. Avian Pathol, 1991,20:233-239.
3．郭玉璞，陈德威，范国雄等．北京小鸭传染性浆膜炎的调查研究[J]．畜牧兽医学报，1982，12(2)：107-112．
4．刘凌云、郑光美主编《普通动物学》(第三版)，高等教育出版社，1997。
5．B．W 卡尔尼克主．高福，苏敬良，主译．禽病学[M]，(第十版)．中国农业出版社，1999，195—20。
6. Sandhu, T.S. immunogenicity and safety of a live pasteurella anatipestifer vaccine in White Pekin ducklings; Laboratory and field trials[J] Avian Pathol, 1991,20:423-432。
7. {Helfer, D.H.Pasteurella anatipestifer infection in Turkeys[J].Avian Dis, 1973,21(4):712-715.
8. Pierce, R.l. and M.W. Vorhies. Pasteurella anatipestifer infection in geese[J].Avian Dis, 1973,17:868-870.
9. Smith, J.M,D.D. Frame, G. Cooper et. al Pasteurella anatipestifer infetion in comm. ercial Meat-type Tokrkeys in California[J].Avian Diseases, 1987,31(5):913-917.
10. Frommer, A.,R. Bock, A. Inbar, and S. Zemer. Muscovy ducks as a source of Pasteurella anatipestifer infection in turkey flocks [J]. Avian Pathol, 1990,19:161-163.
11．蔡文琴，王伯沄主编．实用免疫细胞化学与核酸分子杂交技术．四川科学技术出版社，1994．72-97．4．
12．纪小龙，王淑琴，李红芬．免疫组化在临床诊断中的作用．Chin J Lab Digan, April 1998, Vol 2, No.2．
13. Bisgaard, M Antigenic studies on pasteurella anatipestifer species in certae sedis, Using slide and tube agglutingation[J].Avain pathology, 1982,11:341-350.
14．郭玉璞，甘孟候，张中直．应用荧光抗体法诊断小鸭传染性浆膜炎的实验[J]．中国兽医杂志，1982，8(7)：15-17．
15．苏敬良，黄瑜，吕艳丽．间接荧光抗体技术检测鸭疫里氏杆菌[J]．中国兽医杂志，1999，25(2)：12-13．
16．李桂杰，朱瑞良，郭宝聚．细菌内毒素的研究进展．山东农业大学学报，1999，29
30(2)．
17．许国根，陈突，张万光，等．对于全身炎症反应综合征患者致炎因子与抗炎因子的变化及相关性研究．Chin J Crit Care med june 2002．22(6)．
18．内毒素的产生，(见网页：毒与健康—毒的揭示 2000 见沈阳飞龙制药有限公司，E-mail：webmaster@medr.com)
19．陈君涛，毒素对内皮细胞的损伤机制，第三军医大学西南医院烧伤研究所，Foreign Medical Sciences Section of PathopHysiology．
20．Bevilacqua MP, Paber JS, Wheeler ME, et al. Interleukin 1 acts on cultures haman vascular endothelium to increase the adhesiOn of polymorepHnuclear leukocytes, monocytes and related leukocyte cell lines. J Clin Invest 1985, 76: 2003.
21．陈敏英，管向东．多器官功能障碍综合征的病理生理．中国实用妇科与产科杂志，2002，18(6)．
22．陈建．内毒素几何效应细胞的分子机制，第三军医大学附属新安医院烧伤研究所，重庆，第三军医大学学报，1999，21(11)．
23．陈国强，杨凌，杨永宗．内毒素在动脉粥样硬化形成中的可能作用．衡阳医学院心血管病研究所，生理科学进展，1991，22(4)．
24．陈敏英．多器官功能障碍综合征的病理生理．中国实用妇科与产科杂志，2002，18(6)．
25．许国根，王戈，缪群，等．全身炎症反应综合征患者抗炎因子对T淋巴细胞．凋亡的影响，中华急诊医学杂志，2002，8，11(4)．
26．齐文广，任爱民，张淑文，等．血清促炎、抗炎因子在急性感染患者中的动态变化及中药干预的影响．中国中西医结合杂志，2000，20(11)．
27．夏时海，赵晓晏，郭萍 急性胰腺炎促炎因子和抗炎因子变化的动态观察 第三军医大学附属新桥医院消化内科、重庆侧37第三军医大学学报 2001 23(4))。
28．罗昆仑．嗜中性白细胞介导组织损伤的机理．第三军医大学西南医院肝胆外科，国外医学生理、病理科学与临床分册，1999，19(1)．
29．周向东，陈意生，史景泉，等．内毒素条件下参与嗜中性白细胞—血管内皮细胞粘附过程的分子基础，第三军医大学基础部病理学教研室，第三军医大学学报，1996，8(2)．
30．李春盛，桂培春，何新华，等．NO和iNOS在内毒素诱导的大鼠急性肺损伤的作用极大黄对其影响．中国急救医学，2000，20(2)．
31．汪盛贤，马怖仁，于忠和，等．内毒素性肺损伤大鼠肺表面活性物质的变化．成都军区程度总医院，解放军医学杂志，1997，22(2)．
32．王伟阳．循环内毒素促进肠道内毒素的转运及选择性肠道消毒的影响．病理生理学教研室，北京医科大学学报1993 25(3)．
33．Deitch EA. et al Eadotoxin promotes the transtocation of bacteria from the
gut. Arch Surg 1987: 122:185.
34. O/Dwyer ST, et al.A single dose of endotoxin inzrcases intestinal permeability in healthy humans. Arch Surg 1988;123:1459.
35. Noman JG, Fink G; Frane M et al, Guffay J, Garter G, Dlviyon B, Sexton C, Glaccum M Aruive incerleucin-1 receptor requied for maximal pregression of accte pancrearitis. Am surge,1996;223:163-169.
36. Kimural k, Shimosequece I, Ahe R, Masamine A, Sarah A, Takasa A, Koizumi M, Toyote T. Low dose of lipopolysecchoride upreguiore zccrar cell apoptosis in cenuleir, pancneatitis panxreas, 1998;17:120-126.
37．彭小宁．内毒素诱导淋巴细胞胞凋亡．附属湘雅医院传染科，湖南医科大学学报 2000，25(6)．
38. Negate S.Apoptosis by death faxtor[J]. Cell,1997,28.
39. Barke RA, Roy S .Chapin RB. et al. The induction of accelerated thymic programmed cell death during polymicrobial sepsis: control by corticosceroids but tumor dcriosis factor. Shock. 1995.3:259-267.
40. Parksm A, Bullcy GB, Granger DN, et al.Role of oxygen-free redical in shock, ischemia, and organ preservation. Surgery, 1983,94(3):428-430.
41. PHung C D ,Ezineme T A, Turrens J F, Hydrogen peroxide metabolism in muscle mitochondria, Arch Biochem BiopHys, 1994,315(2),479-482.
42．陆松敏，刘建仓，李萍．毒素休克大鼠肝线粒体质子跨膜转运的改变．第三军医大学野战外科研究所，生物化学与生物物理进展，1999；26(1)．
43．郭德玉，陈意生．实验性内毒素介导的大鼠肾脏病理变化．第三军医大学基础部病理学教研室，第三军医大学学报，1995；17(6)．
44．雄正文，王泊云．影响免疫组织抗原性的因素分析与对策诊断病理学杂志．1997；9：171．
45．熊正文，胡海霞，李春光．盐酸水解暴露抗原促进免疫组织化学反应的研究．中国组织化学与细胞化学杂志，1996；5：71．
46．王伟阳，内毒素血症大鼠肠道内毒素移位的研究，北京医科大学生理学系生理科学进展 1995；26(1)．
47．内源性内毒素移位在失血性休克中的作用及其分子机理研究，第三军医大学外科学 1995；12月．