特异性免疫吸附柱及CRRT清除血浆TNF-α的实验和临床研究
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摘要
多脏器功能障碍综合征(MODS)死亡率极高，对其进行有效治疗，是临床医学的一项重要任务。大量研究证明，系统性炎症反应综合征(SIRS)是MODS动态发展的基础，肿瘤坏死因子-α(TNF-α)是一个在SIRS发病机制中起十分重要作用的关键性前炎症细胞因子。及时有效地清除过度产生的TNF-α，可以抑制SIRS早期细胞因子网络的变化，有助于改善MODS预后。连续性肾脏替代治疗(CRRT)在国内外被广泛地用于救治MODS，因其仅能部分而不是彻底地清除TNF-α，故不能降低MODS的死亡率。吸附疗法具有显著清除TNF-α的功能，但临床应用受到限制。为此，我们从以下两方面进行了研究。第一，应用杂交瘤单克隆抗体技术制备了抗重组人肿瘤坏死因子单克隆抗体，将其包被在载体微球上，制作成功特异性免疫吸附柱，并进行吸附及相应试验。第二，应用CRRT技术治疗14例大器官移植术后MODS患者，ELISA法检测治疗前后血浆TNF-α、IL-1β、IL-6、IL-8浓度，观察清除TNF-α的效果，并将其与特异性吸附柱的吸附效果及优缺点进行综合对比分析。为今后创造出一种将二者有机结合起来治疗MODS的新方法，提供临床及实验依据。
主要结果和结论如下：
1、应用杂交瘤单克隆抗体技术，共建立了9株分泌抗γHTNF-αMcAb的杂交瘤细胞系，继而制备了含McAb的腹水，经免疫亲和层析法得到了纯化的抗rHTNF-αMcAb。
2、特异性交叉反应测定及Western blotting结果显示，这组抗体只与rHTNF具有特异性反应，还具有不同程度的细胞毒中和能力及亲和力。选
取其中特性及功能最佳的单抗，为研制特异性吸附柱奠定了基础。
3、自行设计制作了过滤柱，行硅烷化处理，经XPS检测天N、S、
CI污染，改善了生物相容性。该柱的通透性及实用性能良好，为相关研究
提供了一个新模型。
－4、选用 PSM作为载体微球，依次包被 PLL及抗 rHmF咀 McAb，经
检测包被满意而且牢固。对二次包被的条件进行了探讨。为相关研究提供
了一种新颖的方法。
5、成功制备了特异性兔疫吸附柱，最大吸附率达90％以上。实验发
现吸附存在饱和现象、酸性溶液可洗脱结合抗原，经碱性液缓冲后，仍有
良好的吸附率，证实该柱具有良好的稳定性和重复使用性。创造了一种吸
附疗法中特异性清除TNF－a的新型模式，具有较大的应用价值。
6、常规应用CRRT技术治疗14例大器官移植术后MODS患者，ELISA
法检测治疗前及治疗后不同时间患者血浆TNFo、IL1、IL6及IL毛浓
度。证实了TNFa是大器官移植术后MODS发生发展的关键细胞因子。
提出动态观察TNF－a变化情况，有助于指导临床工作。
7、CRRT能够部分清除此类患者体内TNFa，以吸附机制为主，最大
清除量达44％左右。可以在一定程度上暂时维持患者内环境稳定，但并未
e 明显提高生存率，降低死亡率。
8、TNFo的浓度变化影响了 IL刁 p、IL七及 IL｛的变化，存在正相关
关系。认为早期高效地清除y卜a，将有助于纠正全身性炎症反应，改善
MODS预后。
9、CRRT与特异性吸附柱相比，在治疗MODS中各有优缺点。如将
二者有机地加以结合，将能更加有效地救治MODS患者，改善预后。本研
究为此方面提供了理论及实验依据。
It has been an important task that effective treatment to multiple organ dysfunction syndrome (MODS) for clinical medicine, because of the MODS has extremely high mortality. Many researches have approved that the systemic inflammatory response syndrome (SIRS) was the foundation of the dynamic process of the MODS. Tumor necrosis factor-alpha (TNF- a )is a key pro-inflammatory cytokine which play a very important role in SIRS pathogenesis. Timely and effectively removal tumor necrosis factor-alpha when it was excessive product the changes of cytokine net in the early stage of the systemic inflammatory response syndrome could be suppressed and which is beneficial for improving the prognosis of the multiple organ dysfunction syndrome. Continuous renal replacement therapy (CRRT) had been extensively used for the rescue for the MODS in the world for many years. However, it only partly not totally eliminate the TNF- a and could not depress the mortality of the MODS. Adsorption therapy has obvious function for removal the tumor necrosis factor-alpha, but it was restricted in the clinical application .Fof this reason, we performed the reseach from the two undermentioned aspects.
Firstly, we prepared for the monoclonal antibodies against recombinant human tumor necrosis factor alpha by hybridoma techniques. It was wraped on the polystyrene microsphere carrier connecting poly-L-lysine. After that, we made successfully up the specific immunoadsorption column for the first time,
and took some tests about adsorption. Secondly, we treated to 14 patients with MODS after the big apparatus transplantations by the continous renal replacement therapy techniques. They were detected by ELISA that the concentration of the TNF- a , IL-1 P, IL-6, IL-8 in plasmam before and during the CRRT 1,2,3,6,12 hours respectively.
The main results and conclusions were as follows:
1. Nine groups of hybridoma cell line which can secrete anti-recombinant human tumor necrosis facter alpha were established, and we got some purificative monoclonal antibodies against recombinant human tumor necrosis facter alpha. It grounded for us in whole research.
2. The specific across reaction test and the Western blotting showed that these monoclonal antibodies only reacted specificly to the recombinant human tumor necrosis facter alpha that has 17KDa molecular weight. These McAb possess the neutralization ability for the rHTNF- a cytotoxicity and relative affinity in some degree.
3. The filtration column has favourable capability and practicality, the most penetrating volume was about 2500 milliliters per hour. It provided a new model for some correlative researches.
4.The polystyrene microsphere was chosen for the carrier and coated by poly-L-lysine and anti-recombinant human tumor necrosis facter alpha in turn. Though examining, the two times coating was firm and satisfying. It offered a novel method for this field.
5.We produced triumphantly the specific immunoadsoiption column which the most adsorptive rate was more than 90 percent. This column has good stability and repetition. It created a new pattern of specific eliminate the TNF- a in the adsorption therapy and had biggish application value.
6.There was remarkable running up of the concentration of the TNF- a , IL-1 P , IL-6, IL-8 in plasmm of those patients with MODS after the big
apparatus transplantations.lt was proved that the TNF- a is the key pro-inflammatory cytokine in the pathogenesis of the MODS after translantation.Observing the variety of TNF- a with dynamic mehtod, will help to guidance the clinical work.
7. Partly the TNF- a in these patients could be removaled by CRRT.The main mechanism was the adsorption potentially. It was found that the maximal clearance was gotten for the removal TNF- a by CRRT at 6 hour, and at the same time , the maximal adsorption efficiency was 44%.Although maintaining the circumstance stabilization of the patients temporarily, CRRT could not increase the survival and decrease the mortality in our observation.
8. We substan
主要结果和结论如下：
1、应用杂交瘤单克隆抗体技术，共建立了9株分泌抗γHTNF-αMcAb的杂交瘤细胞系，继而制备了含McAb的腹水，经免疫亲和层析法得到了纯化的抗rHTNF-αMcAb。
2、特异性交叉反应测定及Western blotting结果显示，这组抗体只与rHTNF具有特异性反应，还具有不同程度的细胞毒中和能力及亲和力。选
取其中特性及功能最佳的单抗，为研制特异性吸附柱奠定了基础。
3、自行设计制作了过滤柱，行硅烷化处理，经XPS检测天N、S、
CI污染，改善了生物相容性。该柱的通透性及实用性能良好，为相关研究
提供了一个新模型。
－4、选用 PSM作为载体微球，依次包被 PLL及抗 rHmF咀 McAb，经
检测包被满意而且牢固。对二次包被的条件进行了探讨。为相关研究提供
了一种新颖的方法。
5、成功制备了特异性兔疫吸附柱，最大吸附率达90％以上。实验发
现吸附存在饱和现象、酸性溶液可洗脱结合抗原，经碱性液缓冲后，仍有
良好的吸附率，证实该柱具有良好的稳定性和重复使用性。创造了一种吸
附疗法中特异性清除TNF－a的新型模式，具有较大的应用价值。
6、常规应用CRRT技术治疗14例大器官移植术后MODS患者，ELISA
法检测治疗前及治疗后不同时间患者血浆TNFo、IL1、IL6及IL毛浓
度。证实了TNFa是大器官移植术后MODS发生发展的关键细胞因子。
提出动态观察TNF－a变化情况，有助于指导临床工作。
7、CRRT能够部分清除此类患者体内TNFa，以吸附机制为主，最大
清除量达44％左右。可以在一定程度上暂时维持患者内环境稳定，但并未
e 明显提高生存率，降低死亡率。
8、TNFo的浓度变化影响了 IL刁 p、IL七及 IL｛的变化，存在正相关
关系。认为早期高效地清除y卜a，将有助于纠正全身性炎症反应，改善
MODS预后。
9、CRRT与特异性吸附柱相比，在治疗MODS中各有优缺点。如将
二者有机地加以结合，将能更加有效地救治MODS患者，改善预后。本研
究为此方面提供了理论及实验依据。
It has been an important task that effective treatment to multiple organ dysfunction syndrome (MODS) for clinical medicine, because of the MODS has extremely high mortality. Many researches have approved that the systemic inflammatory response syndrome (SIRS) was the foundation of the dynamic process of the MODS. Tumor necrosis factor-alpha (TNF- a )is a key pro-inflammatory cytokine which play a very important role in SIRS pathogenesis. Timely and effectively removal tumor necrosis factor-alpha when it was excessive product the changes of cytokine net in the early stage of the systemic inflammatory response syndrome could be suppressed and which is beneficial for improving the prognosis of the multiple organ dysfunction syndrome. Continuous renal replacement therapy (CRRT) had been extensively used for the rescue for the MODS in the world for many years. However, it only partly not totally eliminate the TNF- a and could not depress the mortality of the MODS. Adsorption therapy has obvious function for removal the tumor necrosis factor-alpha, but it was restricted in the clinical application .Fof this reason, we performed the reseach from the two undermentioned aspects.
Firstly, we prepared for the monoclonal antibodies against recombinant human tumor necrosis factor alpha by hybridoma techniques. It was wraped on the polystyrene microsphere carrier connecting poly-L-lysine. After that, we made successfully up the specific immunoadsorption column for the first time,
and took some tests about adsorption. Secondly, we treated to 14 patients with MODS after the big apparatus transplantations by the continous renal replacement therapy techniques. They were detected by ELISA that the concentration of the TNF- a , IL-1 P, IL-6, IL-8 in plasmam before and during the CRRT 1,2,3,6,12 hours respectively.
The main results and conclusions were as follows:
1. Nine groups of hybridoma cell line which can secrete anti-recombinant human tumor necrosis facter alpha were established, and we got some purificative monoclonal antibodies against recombinant human tumor necrosis facter alpha. It grounded for us in whole research.
2. The specific across reaction test and the Western blotting showed that these monoclonal antibodies only reacted specificly to the recombinant human tumor necrosis facter alpha that has 17KDa molecular weight. These McAb possess the neutralization ability for the rHTNF- a cytotoxicity and relative affinity in some degree.
3. The filtration column has favourable capability and practicality, the most penetrating volume was about 2500 milliliters per hour. It provided a new model for some correlative researches.
4.The polystyrene microsphere was chosen for the carrier and coated by poly-L-lysine and anti-recombinant human tumor necrosis facter alpha in turn. Though examining, the two times coating was firm and satisfying. It offered a novel method for this field.
5.We produced triumphantly the specific immunoadsoiption column which the most adsorptive rate was more than 90 percent. This column has good stability and repetition. It created a new pattern of specific eliminate the TNF- a in the adsorption therapy and had biggish application value.
6.There was remarkable running up of the concentration of the TNF- a , IL-1 P , IL-6, IL-8 in plasmm of those patients with MODS after the big
apparatus transplantations.lt was proved that the TNF- a is the key pro-inflammatory cytokine in the pathogenesis of the MODS after translantation.Observing the variety of TNF- a with dynamic mehtod, will help to guidance the clinical work.
7. Partly the TNF- a in these patients could be removaled by CRRT.The main mechanism was the adsorption potentially. It was found that the maximal clearance was gotten for the removal TNF- a by CRRT at 6 hour, and at the same time , the maximal adsorption efficiency was 44%.Although maintaining the circumstance stabilization of the patients temporarily, CRRT could not increase the survival and decrease the mortality in our observation.
8. We substan
引文
1．林学颜，张玲主编．现代细胞与分子免疫学，科学出版社 北京，1999，459-488
2．徐宜为编著．免疫检测技术，科学出版社北京，1991，231-301
3．孙卫民，王惠琴编著．细胞因子研究方法学，人民卫生出版社 北京，1999，600-605
4．黎燕，葛学铭，赵薇薇等。抗重组人肿瘤坏死因子-α(rHTNF-α)单克隆抗体特性及功能的研究，单克隆抗体通迅，1991，71(2)：1-4
5. Kohler G, Milstein C. Continuous culture of used cells secreting antibody of predefined specificity. Nature 1975; 256:495～497
6. Fukuyama R. Development and application of a modified monoclonal hybridoma technique for isolating monoclonal antibodies to human brain regains. J Neurosci Methods 1996; 64(1):135～142
7. Yang CC, Chan HL. Neutralizing epitope mapping of six betal-bungarotoxin monoclonal antibodies and its application in betal-bungarotoxin peptide vaccine design. Biochem J 1998; 330:497～503
8. Yu DS, Chang SY, Ma CP. Monoclonal antibodies against renal tumors: the potential application in discrimination of ambiguous adenocarcinoma or transitional cell corcinoma of Kidney. J Urol 1998; 159(1): 48～51
9. Fransens L. Recombinant tumor necrosis factor:its effect and its synergism with interferon on a variety of normol and transformed human cell lines. Eur J Cancer Clin oncol 1986; 22(4): 419～426
10. Fendly BM, Toy KJ, Creasey AA et al. Murine monodonal antibodies defining neutralizing epitopes on tumor necrosis factor. Hybridoma 1987; 6(4): 359～370
11. Katitcioglu SF, Ulus AT, Gokce P et al. Bentificial effects of iloprost
during experimentally induced hemorrhagic shock. Panminerva Med 2000; 42(2): 109～117
12.Herrera Garza E, Cubillos Garzon A, Stetson SJ et al. Tumor necrosis factor-alpha: a mediator in the pathogenesis of cordiac insufficieney. Arch Inst Cardiol Mex 1999; 69(5): 462-468
13.Carins CB, Panacek EA, Harken AH et al. Benck to bedside: tumor necrosis factor-alpha: from inflammation to resuscitation. Acad Emerg Med 2000; 7(8): 930-941
14.Evans TJ. Bioassay for tumor necrosis factor-alpha and beta. Mol Biotechnol 2000; 15(3): 243-248
15.Kraut EJ, Chen S, Hubbard NE et al. Tumor necrosis factor depresses myocardial contractility in endotoxemic swine. J Trauma 1999; 46(5): 900-906
16.Jibiki T, Terai M, Tateno S et al. Expression of tumor necrosis factor-alpha protein in the myocardium in fatal myocarditis. Pediatr Int 2000; 42(1): 43-47
17.Hoffmann JN, Werdan K, Hartl WH et al. Hemofiltrate from patients with severe sepsis and depressed left ventricular contractility contains cardiotoxic componds. Shock 1999; 12(3): 174-180
18.Satoh M, Nakamura M, Satoh H et al. Expression of tumor necrosis factor-alpha-converting enzyme and tumor necrosis factor-alpha in human myocarditis. J Am coll cardiol 2000; 36(4): 1288-1294
19.Yu Y, Si C, Lang Z.The effect of tumor necrosis factor alpha on hepatic necrosis in viral hepatitis,Zhong hua Nei Ke Za Zhi 1996; 35(1): 28～31
20.Liang CM, Liang SM, Jost T et al.Production and characterization of monoclonal antibodies against recombinant human tumor necrosis factor/cachectin. Biochem Biophys Res Commun 1986; 137(2): 847～854
21.Meager A, Parti S, Leung H et al. Preparation and characterization of
monoclonal antibodies directed against antigenic determinants of recombinant human tumour necrosis factor(rTNF). Hybridoma 1987; 6(3): 305～311
22. Guo X, Wang A, Chen S et al. The development of monoclonal antibody against rhTNF and its curative effects on E,coli infected mice. Chin Med Sci J 1997; 12(4): 229～231
23．蔡文琴、王伯云主编。实用免疫细胞化学，成都，四川科学技术出版社 1988，44～56
24. Woodhouse KA, Weitz JI, Brash JL. Interactions of plasminogen and fibrinogen with model silica glass surface:Adsorption from plasma and enzymatic activity studies. J Bio Mat Res 1994; 28(4): 407～415
25．刘彦仿主编．免疫组织化学，北京，人民卫生出版社，1990，27～32
26．胡勇，曾雪梅，公衍道等．用椭圆编术研究血浆白蛋白和纤维蛋白原在硅片及聚氨酯薄膜表面的吸附行为。中国生物医学工程学报 1998；17(1)：38～43
27．肖才德，隋森芳，公衍道等．用SPR生物传感器研究纤维蛋白原在生物医用材料表面的吸附．中国生物医学工程学报 2000；19(1)：60～65
28．俞耀庭主编．生物医用材料，天津，天津大学出版社 2000，12～16
29. Shoffner MA, Cheng J, Hvichia GE et al.Chip PCR.1.Surface passivation of microfabricated silicon-glass chips for PCR. Nucleic Acids Research 1996; 24(2): 375～379
30. Giordano BC, Copeland ER, Landers JP. Towards dynamic coating of glass microchip chambers for amplifying DNA via the polymerase chain reaction. Electrophoresis 2001; 22:334～340
31. Munro NJ, Huhmer AFR, Landers JP. Robust polymeric Microchannel coming for microchip-based analysis of neat PCR products. Anal Chem 2001;1784～1794
32. Rehman FN, Audeh M, Abrams ES et al. Immobilization of
acrylamide-modified digonucleotides by co-polymerization.Nucleic. Acids Research 1999; 27(2): 649～655
33. Cavic BA, McGovern ME, Nisman R et al, High surface density immobilization of oligonucleotide on silicon. Analyst 2001, 126: 485-490
34. Halliwell C, Cass AEG, A factorial analysis of silanization condition for the immobilization of oligonucleotides on glass surfaces. Aral Chem 2001;73:2476-2483
35. Belosludtsev Y, Iverson B, Lemeshko S et al, DNA microarrays basedon noncovalent oligonucleotide attaehment and hybeidization in two dinensions. Anal Biochem 2001; 292:250-256
36. Nashat AH, Moronne M, Ferraril M. Detection of functional geoaps and antibodies on mitrofabeicated surface by confocal microscopy Biotec X Bioeng 1998; 60:137-146
37. Rogers YH, Baucom PJ, Husng ZJ, et al, Immobilization of oligonnclestides onto a glass support via disulfide bonds: a methid for preparation of DNA midroarrays. Anal Biochem 1999; 266:23-30
38. Joos B, Kuster H, Cone R. Covalent attachment of hybridizable oligonucleotidos to glass supports. Anal Biochem 1997; 247:96-101
39．李友华，何应律，赵中一，等．用硅烷化处理改性格硅胶分离富机集痕量元速素．分析化学 1994；22(8)：812—815
40. Coombes AGA, Breeze V, Lin W et al, Laefic acid-stablisheol albumin for microsphere formulation and biomedical coating. Biomaerials 2001; 22(1): 1-8
41. Shymkiw RC, Zernicke RF, Forrester KR et al, Evaluation of laser-Doppler perfusion imaging for measurement of blood flow in cortical bone, J ApplPhysiol 2001; 90(4): 1314-1318
42. Yang CY, Tsay SY, Tsiang RC. Encopsulating aspirin into a sarfactant-free ethyl cellular microsphere using non-toxic solvents by
emulsion solvent-evaporation technigue. J Microencapsul 2001; 18(2): 223-136
43.Iwamaru A, Watanabe M, Yu S et al, Measurement of tumor blood flow using colored dye ertraction microspheres in two rat tumor models. Int J Onlol 2001; 18(2): 227-232
44.Orienti I, Zuccari G, Luppi B et al. Fatty acid substituted potyvinyl alcohal as a supporting material for micrasphere preparation. J Microencapsul 2001; 18(1): 77～87
45.Blohm DH, Guiseppi EA. New developments in microarray technology. Curr Opin Biotechnol 2001; 12(1): 41～47
46.Studinski RC, Vitkin IA. Methodoldgy for reexamining polarized light interactions with tissues and tissuelike media in the exact back scattering direction. J Biomed Opt 2000; 5(3): 330～337
47.Vervact C, Byron PR. Polystyrene microsphere spay standards based on CFC-free inhaler technology. J Aerosol Med 2000; 13(2): 105～115
48.Jacques SL, Roman JR, Leek. Imaging superficial tissues with polarized light. Lasers Surg Med 2000; 26(2): 119～129
49.Kruth HS, Chang J, Ifrim I et al, Characterization of patocytosis endocytosis into macrophage surface-connected compartments. Eur J Cell Biol 1999; 78(2): 91～99
50.Ogawara K, Yoshida M, Higaki K et al. Hepatic uptake of polystytene microspheres in rats:effect of particle size on intrahepatic distribution. J Controlled Relcase 1999; 59(1): 15～22
51.Kempe R, NassH, Fuhrmann H. Lanthanide-mediated ligand transfer reactions---a route to late transition metal bisamido complexes. Chemistry 2001; 7(8): 1630～1636
52.Gorbet MB, Sefton MV. Leukocyte activation and leukocyte procoagulant activities after blood coutact with polystyrene and polyethylene
glycol-immobilized polystyrene beads. J Lab Clim Med 2001; 137(5): 345～355
53. Luo LY, Diamandis EP. Preliminary examination of time-resolved fluorometry for protein array applications. Luminescence 2000; 15(6): 409～413
54. Satok, Tokeshi M, Kimura H et al. Determination of coreinoembryonic autigen in human scra by integrated bead-bed immunoassay in a microchip for cancer diagnosiss. Anal Chem 2001; 73(6): 1213～1218
55. Moss DM, Arrowood MJ. Quantification of Cryptosporidium parvmm oocysts in millse fecal specimens using immunomagnetic particles and two-color flow cytometry. J Parasitol 2001; 87(2): 406～412
56. Zheng N, Carroll SS, Inglese J et al. Miniaturization of a hepatitis C virus PNA polymerase assay using a-102 degrees C cooled CCD camara-based imaging system. Anal Biochem 2001; 290(2): 214～220
57. Ganachaud F, Elacssari A, Pichot C. Covalent grafting of polythy midylic acid outo amine-containing polystyrene latex particles. J Biomater Sci Polym ED 2000; 11(9):931～945
58. Foster KA, Yazdanian M, Audus KL. Microparticulate uptake mechanisms of in-vitro cell culture models of the respiratory epithelium. J Pharm Pharmacol 2001; 53(1): 57～66
59. Coombes AGA, Brecze V, Lin W et al. Lactic acid-stabilised albumin for microsphere formulation and biomedical coatings. Biomaterials 2001; 22(1): 1～8
60. Ogawara K, Yoshida M, Higaki K et al. Hepatic uptake of polystyrene microsphere in rats: effect of particle size on intrahepatic distribution. J Controlled Release 1999; 59(1): 15～22
61．牛杰，陈明哲，李伟联，等．多聚赖氨酸包被球囊导管转移基因表达的时相特征．中国介入心脏病学杂志 1999；7(2)：80～83
62. Asayama S, Maruyama A, Akaike T. Comb-type prepolymers consisting of a polyacrylamide backone and poly(L-Lysine)graft chains for multivalent ligands. Bioconjug Chem 1999; 10(2): 246～253
63. Allo JC, Midoux P, Merten M et al. Efficient gene transfer into human normal and cystic fibrosis tracheal gland serous cells with synthetic vectors. Am J Respir Cell Mol Biol 2000; 22(2): 166～175
64．李均敏，田琣砷，蒋惠秋，等，多聚阳离子对非病毒载体转移效率的影响．中国肿瘤生物治疗杂志 2000；7(1)：6～10
65. Nagaki M, Hughes RD, Keane HM et al. In vitro plasma perfusion through adsorbents and plasma ultrafiltration to remove endotoxin and cytokines. Circulatory Shock 1992; 38:: 182-187
66. Ronco C, Brendolan A, Dan M et al. Absorption in sepsis. Kidney Int 2000; 58(S76): S148-153
67. Tetta C, Bellomo R, Brendolan A et al. Use of adsorptive mechanisms in continuous renal replacement theropies in the critically ill. Kidney Int 1999; 56(S72): S15-19
68. Coraim FJ, Coraim HP, Ebemann R et al. Acutte respiratory failure after cardiac surgery: clinical experience with the application of continous arteriovenous hemofiltration. Crit Care Med 1986: 714～721
69. Cosentino F, Paganin E, Lockrem J et al. Continous arteriovenous hewofiltration in the adult hespiratory distress syndrome.Contrib Nephrol 1991; 9: 94～101
70. Riera JA, Alted E, Iozano MJ et al. Influence of continous hemofiltration on the hemodynamics of trauma patients. Surgery 1997; 122:902～911
71. Tetta C, Cavaillon JM, Schulze MS et al. Removal of cytokine and activated complement componens in an experimental model of continuous plasma filtration coupled with sorbent adsorption. Nephrol Dial Transplant 1998; 13: 1458-1463
72. Reeves JH, Butt WW, Shann F et al. Continuous plasma filtration in sepsis syndrome. Crit Care Med 1999; 27:2096-3001
73. Tetta C, Gianotti L, Cavaillon JM et al. Coupled plasma filtration-adsorption in a rabbit model of endotoxic shock. Crit Care Med 2000; 28: 1526-1533
74．朱云松，侯凡凡，张训．免疫吸附清除内毒素休克兔循环TNF对IL-1水平影响．中华微生物学和免疫学杂志 1999；3：28-30
75. Bane AE. Multiple organ lailure,multiple organ dysfunction syndrome,and 1997; 132: 703～714
76. Beal AL, Cerra FB. Multiple organ failure syndrome in the 1990-systemic inflammatory response and organ dysfunction. JAMA 1994; 271: 226～236
77. Bone R. Immunologic dissonance: A continuing evolution in our understanding of the systemic inflammatory response syndrome (SIRS) and the multiple organ dysfunction syndrome (MODS). Ann Intern Med 1996; 125:680-688
78. Mrshima S, Yukioka T, Matsuda H et al. Mild hypotension and body burns synergistically increase bacterial translocations in rats consistent with a"two-hit phenomenon". J Burn Care Rehabil 1997; 18:22～31
79. Yao YM, Sheng ZY, Yu Y et al. The potential etiologic role of tumor necrosis faltor in mediating multiple organ dysfunction in rats following intestinal ischemia-reperfusion injury. Resuscitation 1995; 29: 57～161
80. Hu S, Sheng ZY, Zhou BT et al. Study on delay two-phase multiple organ dysfunction syndrome. Chin Med J 1998; 111: 101～105
81. Henas FR, Daes JE, Dennis FJ. Risk factors for multiple organ failure:a case control study. J Trauma 1991; 31: 74～77
82. Rangel-Fausto MS, Pitter D, Coutig M et al.The mature history of the systemic inflammatory response syndrome. JAMA 1995; 273: 117～126
83. Yao YM, Redl H, Bahrami S et al. The inflammatory basis of trauma/shock
associated multiple organ failure. Inflamm Res 1998; 47: 201～210
84.Marshall JG. Clinical trails of mediator-directed therapy in sepsis: What have we learned? Intensive Care Med 2000; 26:S75-81
85.Bone R, Groding C, Bald R. Sepsis: A new hypothesis for the disease process. Chest 1997, 112:235-241
86.Koch T. Origin and mediators involved in sepsis and the systemic inflammatory resonse syndrome. Kidney Int 1998; 53(S64): S66-72
87.Ayala A, Kart SM, Evans TA et al. Factors responsible for peritoneal granulocyte apoptosis oluring sepsis. J Surg Res 1997; 69(1): 67-75
88.Schumann RR, Belka C, Reuter D et al. Lipopolysaccharide activate caspasel (interleukin-converting enzyme) in cultured monocytic and endothelial cells. Blood 1998; 91(2); 577-584
89.Ford H, Watkins S, Reblock K et al. The role of inflammatory cytokines and nitric oxide in the pathogenesis of necrotizing enterocolitis. J Pediatr Surg 1997; 32(2): 275-282
90.Kelly JL, O'Sullivan C, O'Riordain M et al. Is circulating endotoxin the trigger for the systemic inflammatory response syndrome scen after injury?Ann Surg 1997; 225: 530～538
91.Brauner JS, Rohde LE, Clausell N. Circulating endothelin-1 and tumor necrosis factor-alpha: early predictors of mortality in patients with septic shock.. Intensive Care Med 2000; 26(3): 305-313
92.Herrera Garza E, Cubillos Garzon A, Stetson SJ et al. Tumor necrosis factor-alpha: a mediator in the pathogenesis of cordiac insufficieney. Arch Inst Cardiol Mex 1999; 69(5): 462-468
93.Katitcioglu SF, Ulus AT, Gokce P et al. Benificial effects of iloprost during experimentally induced hemorrhagic shock.. Panminerva Med 2000; 42(2): 109-117
94.Carins CB, Panacek EA, Harken AH et al. Benck to bedside: tumor
necrosis factor-alpha: from inflammation to resuscitation. Acad Emerg Med 2000; 7(8): 930-941
95. Evans TJ. Bioassay for tumor necrosis factor-alpha and beta. Mol Biotechnol 2000; 15(3); 243-248
96. Bellomo R, Kellum JA, Gandhi CR et al. The effect of intensive plasma water exchange by hemofiltration on hemodynamics and soluble mediators in canine endotoxemia. Am J Respir Crit Care Med 2000; 161(5): 1429-1436
97. Journois D. Hemofillration during cardcopulmonary bypass. Kidney In 1998; 53(Suppl 66): 174～177
98. Heiolemann S, Ofenstein JP, Sarnaik AP et al. Efficacy of continous arteriovenous hemofiltration in endotoxic shock. Cir Shock 1994; 44: 183～187
99. Singer M, Mcnally T, Screaton G et al. Heparin clearance during continous veno-venous hemofiltration. Intensiw Care Med 1994; 20: 212～215
100 Forni LG, Hilton PJ. Continous hemofiltration in the treatment of acute remal failure. N Engl J Med 1997; 336: 1303～1308
101 Vincent JL, Tielemans C. Continous hemofiltration in severe sepsis:is it beneficial? J Crit Care 1995; 10: 27～32
102 Webb AR, Mythen MG, Mackie IJ. Maintaining blood flow in the extracorporeal circuit:haemostasis and anticoagulation. Intensive Care Med 1995; 21: 84～93
103 Lee P, Weger G, Pryor R et al. Effects of flter pore size on efficacy of continuous arteriovenous hemofiltration therapy for staphylococcus aureus-induced septicemia in immature swine. Crit Care Med 1998; 26: 730-737
104 Rogiers P, Zhang H, Smail N et al. Continuous venovenous hemofiltrution improves cardiac performance by mechanisms other than tumor necrosis
factor-alpha attentuation during endotoxic shock. Crit Care Med 1999; 27(9): 1848-1855
105 De Vfiese AS, Colardyn FA, Philippe JJ et al. Cytokine removal during continuous hemofiltration in septic patients. J Am Soc Nephol 1999; 10(4): 846-853
106 Bellomo R, Kellum JA, Gandhi CR et al. The effect of intensive plasma water exchange by hemofiltration on hemodynamics and soluble mediators in canine endotoxemia. Am J Respir Crit Care Med 2000; 161(5): 1429-1436
107 Lonnemann G, Floege J, Kliem V et al. Extended daily veno-venous high-flux haemodialysis in patients with acute renal failure and multiple organ dysfunction syndrome using a single path batch dialysis system. Nephrol Dial Transplant 2000; 15: 1189-1193
108 Druml W. Metabolic aspects of continuous renal replacement therapies. Kidney Int 1999; 56(S72): S56-61
2．徐宜为编著．免疫检测技术，科学出版社北京，1991，231-301
3．孙卫民，王惠琴编著．细胞因子研究方法学，人民卫生出版社 北京，1999，600-605
4．黎燕，葛学铭，赵薇薇等。抗重组人肿瘤坏死因子-α(rHTNF-α)单克隆抗体特性及功能的研究，单克隆抗体通迅，1991，71(2)：1-4
5. Kohler G, Milstein C. Continuous culture of used cells secreting antibody of predefined specificity. Nature 1975; 256:495～497
6. Fukuyama R. Development and application of a modified monoclonal hybridoma technique for isolating monoclonal antibodies to human brain regains. J Neurosci Methods 1996; 64(1):135～142
7. Yang CC, Chan HL. Neutralizing epitope mapping of six betal-bungarotoxin monoclonal antibodies and its application in betal-bungarotoxin peptide vaccine design. Biochem J 1998; 330:497～503
8. Yu DS, Chang SY, Ma CP. Monoclonal antibodies against renal tumors: the potential application in discrimination of ambiguous adenocarcinoma or transitional cell corcinoma of Kidney. J Urol 1998; 159(1): 48～51
9. Fransens L. Recombinant tumor necrosis factor:its effect and its synergism with interferon on a variety of normol and transformed human cell lines. Eur J Cancer Clin oncol 1986; 22(4): 419～426
10. Fendly BM, Toy KJ, Creasey AA et al. Murine monodonal antibodies defining neutralizing epitopes on tumor necrosis factor. Hybridoma 1987; 6(4): 359～370
11. Katitcioglu SF, Ulus AT, Gokce P et al. Bentificial effects of iloprost
during experimentally induced hemorrhagic shock. Panminerva Med 2000; 42(2): 109～117
12.Herrera Garza E, Cubillos Garzon A, Stetson SJ et al. Tumor necrosis factor-alpha: a mediator in the pathogenesis of cordiac insufficieney. Arch Inst Cardiol Mex 1999; 69(5): 462-468
13.Carins CB, Panacek EA, Harken AH et al. Benck to bedside: tumor necrosis factor-alpha: from inflammation to resuscitation. Acad Emerg Med 2000; 7(8): 930-941
14.Evans TJ. Bioassay for tumor necrosis factor-alpha and beta. Mol Biotechnol 2000; 15(3): 243-248
15.Kraut EJ, Chen S, Hubbard NE et al. Tumor necrosis factor depresses myocardial contractility in endotoxemic swine. J Trauma 1999; 46(5): 900-906
16.Jibiki T, Terai M, Tateno S et al. Expression of tumor necrosis factor-alpha protein in the myocardium in fatal myocarditis. Pediatr Int 2000; 42(1): 43-47
17.Hoffmann JN, Werdan K, Hartl WH et al. Hemofiltrate from patients with severe sepsis and depressed left ventricular contractility contains cardiotoxic componds. Shock 1999; 12(3): 174-180
18.Satoh M, Nakamura M, Satoh H et al. Expression of tumor necrosis factor-alpha-converting enzyme and tumor necrosis factor-alpha in human myocarditis. J Am coll cardiol 2000; 36(4): 1288-1294
19.Yu Y, Si C, Lang Z.The effect of tumor necrosis factor alpha on hepatic necrosis in viral hepatitis,Zhong hua Nei Ke Za Zhi 1996; 35(1): 28～31
20.Liang CM, Liang SM, Jost T et al.Production and characterization of monoclonal antibodies against recombinant human tumor necrosis factor/cachectin. Biochem Biophys Res Commun 1986; 137(2): 847～854
21.Meager A, Parti S, Leung H et al. Preparation and characterization of
monoclonal antibodies directed against antigenic determinants of recombinant human tumour necrosis factor(rTNF). Hybridoma 1987; 6(3): 305～311
22. Guo X, Wang A, Chen S et al. The development of monoclonal antibody against rhTNF and its curative effects on E,coli infected mice. Chin Med Sci J 1997; 12(4): 229～231
23．蔡文琴、王伯云主编。实用免疫细胞化学，成都，四川科学技术出版社 1988，44～56
24. Woodhouse KA, Weitz JI, Brash JL. Interactions of plasminogen and fibrinogen with model silica glass surface:Adsorption from plasma and enzymatic activity studies. J Bio Mat Res 1994; 28(4): 407～415
25．刘彦仿主编．免疫组织化学，北京，人民卫生出版社，1990，27～32
26．胡勇，曾雪梅，公衍道等．用椭圆编术研究血浆白蛋白和纤维蛋白原在硅片及聚氨酯薄膜表面的吸附行为。中国生物医学工程学报 1998；17(1)：38～43
27．肖才德，隋森芳，公衍道等．用SPR生物传感器研究纤维蛋白原在生物医用材料表面的吸附．中国生物医学工程学报 2000；19(1)：60～65
28．俞耀庭主编．生物医用材料，天津，天津大学出版社 2000，12～16
29. Shoffner MA, Cheng J, Hvichia GE et al.Chip PCR.1.Surface passivation of microfabricated silicon-glass chips for PCR. Nucleic Acids Research 1996; 24(2): 375～379
30. Giordano BC, Copeland ER, Landers JP. Towards dynamic coating of glass microchip chambers for amplifying DNA via the polymerase chain reaction. Electrophoresis 2001; 22:334～340
31. Munro NJ, Huhmer AFR, Landers JP. Robust polymeric Microchannel coming for microchip-based analysis of neat PCR products. Anal Chem 2001;1784～1794
32. Rehman FN, Audeh M, Abrams ES et al. Immobilization of
acrylamide-modified digonucleotides by co-polymerization.Nucleic. Acids Research 1999; 27(2): 649～655
33. Cavic BA, McGovern ME, Nisman R et al, High surface density immobilization of oligonucleotide on silicon. Analyst 2001, 126: 485-490
34. Halliwell C, Cass AEG, A factorial analysis of silanization condition for the immobilization of oligonucleotides on glass surfaces. Aral Chem 2001;73:2476-2483
35. Belosludtsev Y, Iverson B, Lemeshko S et al, DNA microarrays basedon noncovalent oligonucleotide attaehment and hybeidization in two dinensions. Anal Biochem 2001; 292:250-256
36. Nashat AH, Moronne M, Ferraril M. Detection of functional geoaps and antibodies on mitrofabeicated surface by confocal microscopy Biotec X Bioeng 1998; 60:137-146
37. Rogers YH, Baucom PJ, Husng ZJ, et al, Immobilization of oligonnclestides onto a glass support via disulfide bonds: a methid for preparation of DNA midroarrays. Anal Biochem 1999; 266:23-30
38. Joos B, Kuster H, Cone R. Covalent attachment of hybridizable oligonucleotidos to glass supports. Anal Biochem 1997; 247:96-101
39．李友华，何应律，赵中一，等．用硅烷化处理改性格硅胶分离富机集痕量元速素．分析化学 1994；22(8)：812—815
40. Coombes AGA, Breeze V, Lin W et al, Laefic acid-stablisheol albumin for microsphere formulation and biomedical coating. Biomaerials 2001; 22(1): 1-8
41. Shymkiw RC, Zernicke RF, Forrester KR et al, Evaluation of laser-Doppler perfusion imaging for measurement of blood flow in cortical bone, J ApplPhysiol 2001; 90(4): 1314-1318
42. Yang CY, Tsay SY, Tsiang RC. Encopsulating aspirin into a sarfactant-free ethyl cellular microsphere using non-toxic solvents by
emulsion solvent-evaporation technigue. J Microencapsul 2001; 18(2): 223-136
43.Iwamaru A, Watanabe M, Yu S et al, Measurement of tumor blood flow using colored dye ertraction microspheres in two rat tumor models. Int J Onlol 2001; 18(2): 227-232
44.Orienti I, Zuccari G, Luppi B et al. Fatty acid substituted potyvinyl alcohal as a supporting material for micrasphere preparation. J Microencapsul 2001; 18(1): 77～87
45.Blohm DH, Guiseppi EA. New developments in microarray technology. Curr Opin Biotechnol 2001; 12(1): 41～47
46.Studinski RC, Vitkin IA. Methodoldgy for reexamining polarized light interactions with tissues and tissuelike media in the exact back scattering direction. J Biomed Opt 2000; 5(3): 330～337
47.Vervact C, Byron PR. Polystyrene microsphere spay standards based on CFC-free inhaler technology. J Aerosol Med 2000; 13(2): 105～115
48.Jacques SL, Roman JR, Leek. Imaging superficial tissues with polarized light. Lasers Surg Med 2000; 26(2): 119～129
49.Kruth HS, Chang J, Ifrim I et al, Characterization of patocytosis endocytosis into macrophage surface-connected compartments. Eur J Cell Biol 1999; 78(2): 91～99
50.Ogawara K, Yoshida M, Higaki K et al. Hepatic uptake of polystytene microspheres in rats:effect of particle size on intrahepatic distribution. J Controlled Relcase 1999; 59(1): 15～22
51.Kempe R, NassH, Fuhrmann H. Lanthanide-mediated ligand transfer reactions---a route to late transition metal bisamido complexes. Chemistry 2001; 7(8): 1630～1636
52.Gorbet MB, Sefton MV. Leukocyte activation and leukocyte procoagulant activities after blood coutact with polystyrene and polyethylene
glycol-immobilized polystyrene beads. J Lab Clim Med 2001; 137(5): 345～355
53. Luo LY, Diamandis EP. Preliminary examination of time-resolved fluorometry for protein array applications. Luminescence 2000; 15(6): 409～413
54. Satok, Tokeshi M, Kimura H et al. Determination of coreinoembryonic autigen in human scra by integrated bead-bed immunoassay in a microchip for cancer diagnosiss. Anal Chem 2001; 73(6): 1213～1218
55. Moss DM, Arrowood MJ. Quantification of Cryptosporidium parvmm oocysts in millse fecal specimens using immunomagnetic particles and two-color flow cytometry. J Parasitol 2001; 87(2): 406～412
56. Zheng N, Carroll SS, Inglese J et al. Miniaturization of a hepatitis C virus PNA polymerase assay using a-102 degrees C cooled CCD camara-based imaging system. Anal Biochem 2001; 290(2): 214～220
57. Ganachaud F, Elacssari A, Pichot C. Covalent grafting of polythy midylic acid outo amine-containing polystyrene latex particles. J Biomater Sci Polym ED 2000; 11(9):931～945
58. Foster KA, Yazdanian M, Audus KL. Microparticulate uptake mechanisms of in-vitro cell culture models of the respiratory epithelium. J Pharm Pharmacol 2001; 53(1): 57～66
59. Coombes AGA, Brecze V, Lin W et al. Lactic acid-stabilised albumin for microsphere formulation and biomedical coatings. Biomaterials 2001; 22(1): 1～8
60. Ogawara K, Yoshida M, Higaki K et al. Hepatic uptake of polystyrene microsphere in rats: effect of particle size on intrahepatic distribution. J Controlled Release 1999; 59(1): 15～22
61．牛杰，陈明哲，李伟联，等．多聚赖氨酸包被球囊导管转移基因表达的时相特征．中国介入心脏病学杂志 1999；7(2)：80～83
62. Asayama S, Maruyama A, Akaike T. Comb-type prepolymers consisting of a polyacrylamide backone and poly(L-Lysine)graft chains for multivalent ligands. Bioconjug Chem 1999; 10(2): 246～253
63. Allo JC, Midoux P, Merten M et al. Efficient gene transfer into human normal and cystic fibrosis tracheal gland serous cells with synthetic vectors. Am J Respir Cell Mol Biol 2000; 22(2): 166～175
64．李均敏，田琣砷，蒋惠秋，等，多聚阳离子对非病毒载体转移效率的影响．中国肿瘤生物治疗杂志 2000；7(1)：6～10
65. Nagaki M, Hughes RD, Keane HM et al. In vitro plasma perfusion through adsorbents and plasma ultrafiltration to remove endotoxin and cytokines. Circulatory Shock 1992; 38:: 182-187
66. Ronco C, Brendolan A, Dan M et al. Absorption in sepsis. Kidney Int 2000; 58(S76): S148-153
67. Tetta C, Bellomo R, Brendolan A et al. Use of adsorptive mechanisms in continuous renal replacement theropies in the critically ill. Kidney Int 1999; 56(S72): S15-19
68. Coraim FJ, Coraim HP, Ebemann R et al. Acutte respiratory failure after cardiac surgery: clinical experience with the application of continous arteriovenous hemofiltration. Crit Care Med 1986: 714～721
69. Cosentino F, Paganin E, Lockrem J et al. Continous arteriovenous hewofiltration in the adult hespiratory distress syndrome.Contrib Nephrol 1991; 9: 94～101
70. Riera JA, Alted E, Iozano MJ et al. Influence of continous hemofiltration on the hemodynamics of trauma patients. Surgery 1997; 122:902～911
71. Tetta C, Cavaillon JM, Schulze MS et al. Removal of cytokine and activated complement componens in an experimental model of continuous plasma filtration coupled with sorbent adsorption. Nephrol Dial Transplant 1998; 13: 1458-1463
72. Reeves JH, Butt WW, Shann F et al. Continuous plasma filtration in sepsis syndrome. Crit Care Med 1999; 27:2096-3001
73. Tetta C, Gianotti L, Cavaillon JM et al. Coupled plasma filtration-adsorption in a rabbit model of endotoxic shock. Crit Care Med 2000; 28: 1526-1533
74．朱云松，侯凡凡，张训．免疫吸附清除内毒素休克兔循环TNF对IL-1水平影响．中华微生物学和免疫学杂志 1999；3：28-30
75. Bane AE. Multiple organ lailure,multiple organ dysfunction syndrome,and 1997; 132: 703～714
76. Beal AL, Cerra FB. Multiple organ failure syndrome in the 1990-systemic inflammatory response and organ dysfunction. JAMA 1994; 271: 226～236
77. Bone R. Immunologic dissonance: A continuing evolution in our understanding of the systemic inflammatory response syndrome (SIRS) and the multiple organ dysfunction syndrome (MODS). Ann Intern Med 1996; 125:680-688
78. Mrshima S, Yukioka T, Matsuda H et al. Mild hypotension and body burns synergistically increase bacterial translocations in rats consistent with a"two-hit phenomenon". J Burn Care Rehabil 1997; 18:22～31
79. Yao YM, Sheng ZY, Yu Y et al. The potential etiologic role of tumor necrosis faltor in mediating multiple organ dysfunction in rats following intestinal ischemia-reperfusion injury. Resuscitation 1995; 29: 57～161
80. Hu S, Sheng ZY, Zhou BT et al. Study on delay two-phase multiple organ dysfunction syndrome. Chin Med J 1998; 111: 101～105
81. Henas FR, Daes JE, Dennis FJ. Risk factors for multiple organ failure:a case control study. J Trauma 1991; 31: 74～77
82. Rangel-Fausto MS, Pitter D, Coutig M et al.The mature history of the systemic inflammatory response syndrome. JAMA 1995; 273: 117～126
83. Yao YM, Redl H, Bahrami S et al. The inflammatory basis of trauma/shock
associated multiple organ failure. Inflamm Res 1998; 47: 201～210
84.Marshall JG. Clinical trails of mediator-directed therapy in sepsis: What have we learned? Intensive Care Med 2000; 26:S75-81
85.Bone R, Groding C, Bald R. Sepsis: A new hypothesis for the disease process. Chest 1997, 112:235-241
86.Koch T. Origin and mediators involved in sepsis and the systemic inflammatory resonse syndrome. Kidney Int 1998; 53(S64): S66-72
87.Ayala A, Kart SM, Evans TA et al. Factors responsible for peritoneal granulocyte apoptosis oluring sepsis. J Surg Res 1997; 69(1): 67-75
88.Schumann RR, Belka C, Reuter D et al. Lipopolysaccharide activate caspasel (interleukin-converting enzyme) in cultured monocytic and endothelial cells. Blood 1998; 91(2); 577-584
89.Ford H, Watkins S, Reblock K et al. The role of inflammatory cytokines and nitric oxide in the pathogenesis of necrotizing enterocolitis. J Pediatr Surg 1997; 32(2): 275-282
90.Kelly JL, O'Sullivan C, O'Riordain M et al. Is circulating endotoxin the trigger for the systemic inflammatory response syndrome scen after injury?Ann Surg 1997; 225: 530～538
91.Brauner JS, Rohde LE, Clausell N. Circulating endothelin-1 and tumor necrosis factor-alpha: early predictors of mortality in patients with septic shock.. Intensive Care Med 2000; 26(3): 305-313
92.Herrera Garza E, Cubillos Garzon A, Stetson SJ et al. Tumor necrosis factor-alpha: a mediator in the pathogenesis of cordiac insufficieney. Arch Inst Cardiol Mex 1999; 69(5): 462-468
93.Katitcioglu SF, Ulus AT, Gokce P et al. Benificial effects of iloprost during experimentally induced hemorrhagic shock.. Panminerva Med 2000; 42(2): 109-117
94.Carins CB, Panacek EA, Harken AH et al. Benck to bedside: tumor
necrosis factor-alpha: from inflammation to resuscitation. Acad Emerg Med 2000; 7(8): 930-941
95. Evans TJ. Bioassay for tumor necrosis factor-alpha and beta. Mol Biotechnol 2000; 15(3); 243-248
96. Bellomo R, Kellum JA, Gandhi CR et al. The effect of intensive plasma water exchange by hemofiltration on hemodynamics and soluble mediators in canine endotoxemia. Am J Respir Crit Care Med 2000; 161(5): 1429-1436
97. Journois D. Hemofillration during cardcopulmonary bypass. Kidney In 1998; 53(Suppl 66): 174～177
98. Heiolemann S, Ofenstein JP, Sarnaik AP et al. Efficacy of continous arteriovenous hemofiltration in endotoxic shock. Cir Shock 1994; 44: 183～187
99. Singer M, Mcnally T, Screaton G et al. Heparin clearance during continous veno-venous hemofiltration. Intensiw Care Med 1994; 20: 212～215
100 Forni LG, Hilton PJ. Continous hemofiltration in the treatment of acute remal failure. N Engl J Med 1997; 336: 1303～1308
101 Vincent JL, Tielemans C. Continous hemofiltration in severe sepsis:is it beneficial? J Crit Care 1995; 10: 27～32
102 Webb AR, Mythen MG, Mackie IJ. Maintaining blood flow in the extracorporeal circuit:haemostasis and anticoagulation. Intensive Care Med 1995; 21: 84～93
103 Lee P, Weger G, Pryor R et al. Effects of flter pore size on efficacy of continuous arteriovenous hemofiltration therapy for staphylococcus aureus-induced septicemia in immature swine. Crit Care Med 1998; 26: 730-737
104 Rogiers P, Zhang H, Smail N et al. Continuous venovenous hemofiltrution improves cardiac performance by mechanisms other than tumor necrosis
factor-alpha attentuation during endotoxic shock. Crit Care Med 1999; 27(9): 1848-1855
105 De Vfiese AS, Colardyn FA, Philippe JJ et al. Cytokine removal during continuous hemofiltration in septic patients. J Am Soc Nephol 1999; 10(4): 846-853
106 Bellomo R, Kellum JA, Gandhi CR et al. The effect of intensive plasma water exchange by hemofiltration on hemodynamics and soluble mediators in canine endotoxemia. Am J Respir Crit Care Med 2000; 161(5): 1429-1436
107 Lonnemann G, Floege J, Kliem V et al. Extended daily veno-venous high-flux haemodialysis in patients with acute renal failure and multiple organ dysfunction syndrome using a single path batch dialysis system. Nephrol Dial Transplant 2000; 15: 1189-1193
108 Druml W. Metabolic aspects of continuous renal replacement therapies. Kidney Int 1999; 56(S72): S56-61