核仁组合成区相关嗜银蛋白对恶性肿瘤诊断的研究
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摘要
肿瘤的发生与机体的免疫状态是密切相关的。事实上正常人机体每天都有10~7-10~9个细胞发生基因突变，并产生恶性表型的瘤细胞，但一般并不会发生肿瘤。很重要的原因就在于机体的免疫监视系统能清除这些突变细胞。当肿瘤细胞的发生超出免疫监视时，就可形成肿瘤。
核仁组成区相关嗜银蛋白(Ag-NORs)是一种酸性非组蛋白，在rDNA转录活性中起重要调控作用，可用一步银染方法显示。银染强度反映了蛋白质合成的能力，与细胞增殖密切相关。核仁组成区相关嗜银蛋白在肿瘤中的研究已有很多报道，认为其与肿瘤增殖细胞核抗原PCNA、Ki-67密切相关。与肿瘤的良恶性鉴别诊断密切相关，与肿瘤的癌前病变相关，与肿瘤的分级、分期密切相关。
T淋巴细胞是机体参与抗肿瘤免疫应答的主要免疫活性细胞，它不仅是细胞免疫的主要效应细胞，而且更重要的是免疫调节细胞。其免疫调节是通过其亚群细胞实现的。
红细胞表面有许多与免疫有关的物质，如CR_1、CR_3、CD_58、CD_59、DAF、SOD酶等均参与机体复杂的免疫网络调控作用。红细胞具有免疫粘附功能，能粘附、运输、清除循环系统免疫复合物，并参与T、B、NK、LAK细胞及
中国人民解放军第四军医大学硕士学位论文
吞噬细胞免疫活性的调控，是机体抗肿瘤免疫的一个重要组成部分。
本实验采用 AgNORS图像分析定量测定方法、T淋巴细胞亚群检测试剂
盒（单克隆抗体直接免疫荧光染色法）、红细胞花环试验对86例恶性肿瘤患
者和42例正常对照组分别测定其外周血T淋巴细胞核仁银染区面积／细胞核
面积比值（IS％）、T 淋巴细胞亚群、红细胞免疫 C3b 受体花环率
（RBC（力）与红细胞循环免疫复合物花环率（RBCJCR卜结果显示：门）
肿瘤组 1＊％值减低，与 42例正常对照组比较差异十分显著少0刀 1）；u）肝
细胞癌组 L S％值的变化与临床分期密切相关，与病理分型无关；（3）肝细胞
癌组ISO／O值变化与肝硬化组，正常组有显著差异 V＜001），肝硬化组与正
常组之间有显著差异（＜0刀5）；u）肝细胞癌组1S％指标变化与淋巳结是否
有转移密切相关；（5）肿瘤组CD3”、CD4”、CD4－／CDS”比值减低，与正常对
照组比较差异十分显著（P叼01人＊仇－值与正常对照组比较无显著意义什
＞00s）；u）肿瘤组RBOQ＊g明显降低，RBC1＊R明显增高，与正常组比
较有显著差异（P＜001h（7）肿瘤组二＊％值与＊o丫0／比值呈正相关
（rro刀3民P＜0 01）；二＊％值与Q＊RR呈正相关（r＝0 503，P＜ o刀1）。
恶性肿瘤患者 IS％值减低，表明机体淋巴细胞增殖活性明显减低，I．S％
值是反映细胞增殖的敏感指标。恶性肿瘤患者CD。－／CD。－比值减低，表明T
细胞介导的细胞免疫功能降低和平衡失调，RBC－C力M降低，RBC－ICR增
高，提示RBC－C巾受体沾性减低及机体清除IC的能力减低。上述作用的结
乏 果是导致整个机体免疫系统功能降低，削弱了抗肿瘤能力。
本研究提示：图像分析法检测肿瘤患者外周血T淋巴细胞中核仁组成区
相关嗜银蛋白强度与机体免疫功能状态密切相关，对肿瘤患者的临床诊断，
治疗监测和判断预后提供一个新的简便可靠的方怯，在临床应用方面有重要
意义。
The pathogenesis of tumour is associated intimately with body s immune state.In fact, in normal body, 107-109 cells are undergoing mutation everyday. Tumour cells with malignant epitope are produced but usually tumour doesri t appear. An important reason is that the body s immunosurreillance can eliminate these cells. When the increase of tumour cells surpass the limit of immunosurreillance, tumour will be formed.
Bing responsible for transcriptional actively, the argyrophilic nuclolar regions (Ag-NORs) are acidic nonhistone proteins and are identified by a one-step silver staining technique. The argyrophilia appears to reflect rDNA transcription activity, which has been correlated with proliferative activity of cells. There are many studies about Ag-NORs on tumours, indicated that was correlation with
PCNA, Ki-67; benign and malignant tumour; potentially malignant lesions;grade of tumour.
T lymphocytes are the major immuno-active cells, which participate in the immunoreaction against tumour. Not only do they play a great role in cell immunity, but also do they play role in immunoregulation. The regulation is realized by its subgroup cells.
On the surface of red cell, there are many substances participating in the immunoreaction, such as CR^ CRs> CDs8> CD.s9> DAF^ SOD enzyme,etc. They all take part in the complex regulation of immunonetwork. Red cells have the function of immune adherence. They can adhere, transport and eliminate immune complex in circulation. They play role in the regulation of immune activity of T, B> NIC-. LAK cells and phagocytes, too. So red cells are an important part in the body' s immunity against tumour.
Nucleolar organizer regions and T lymphocyte subgroups (including CD3 , CD4*> CDg")of peripheral blood and erythrocyte immune function (including RBC-CsbRR and RBC-ICR) were measured in 86 patients with malignant tumours (such as liver,osteoma ,esophagus).The methods of Ag-NORs KL image-analysis,direct immunoflurescent staining technique with flow cytometry(FCM) and immune adherence rosette were used in the study.The results showed that:(l)The value of I.S% was lower in cancer patients than those of 42 healthy control subjects (PO.01). (2) The variation of I.S% ratio was significantly relative with clinical stages and was not relative with pathological classification in liver. (3) The value of I.S% was lower in liver patients than those of 22 cirrhosis and 42 healthy control subjects (P<0.01) and was stitistical different between cirrhosis and healthy control (P<0.05). (4) The value of I.S% was significantly relative with tumour transformation.^) The values of CDs*, CD4", CD47CDs~
were lower in cancer patients than those of 42 healthy control subjects (P<0.01) and the value of CDg+ was not statistical different between two groups CP>0.05).The values of RBC-C3bRR was lower and RBC-ICR was higher in the group of cancer patients as compared with the group of 42 healthy control subjects (P<0.01). (7)The value of I.S% significantly correlated with the CD4 /CD8~ ratio in the cancer groupe (r=0.713, PO.01) and the value of I.S% significantly correlated with the value of RBC-C3bRR in cancer groupe(r= 0.503, P<0.01).
The decreased I.S% ratio indicated that the cell proliferation was depressed. The I.S% ratio was a sensitive index of reflecting cell proliferation.The decreased CD4+/CDg^ ratio indicated by T lymphocyte was depresed and balance was interrupted. The decreased value of RBC-C3bRR and increased RBC-ICR demonstrated that RBC-Csb receptor activity and 1C elimination was significantly depressed in the patients with malignant tumours. The above effects result in the whole immune system function and the anti-tumour-immune activity depressed.
The Index I.S% significantly correlated with the immune function of body. The detected of I.S% ratio has important significance in clinical diagnosis and threapy evalueated and prognosis assessment of malignat tumourand a simple and exact methed.
核仁组成区相关嗜银蛋白(Ag-NORs)是一种酸性非组蛋白，在rDNA转录活性中起重要调控作用，可用一步银染方法显示。银染强度反映了蛋白质合成的能力，与细胞增殖密切相关。核仁组成区相关嗜银蛋白在肿瘤中的研究已有很多报道，认为其与肿瘤增殖细胞核抗原PCNA、Ki-67密切相关。与肿瘤的良恶性鉴别诊断密切相关，与肿瘤的癌前病变相关，与肿瘤的分级、分期密切相关。
T淋巴细胞是机体参与抗肿瘤免疫应答的主要免疫活性细胞，它不仅是细胞免疫的主要效应细胞，而且更重要的是免疫调节细胞。其免疫调节是通过其亚群细胞实现的。
红细胞表面有许多与免疫有关的物质，如CR_1、CR_3、CD_58、CD_59、DAF、SOD酶等均参与机体复杂的免疫网络调控作用。红细胞具有免疫粘附功能，能粘附、运输、清除循环系统免疫复合物，并参与T、B、NK、LAK细胞及
中国人民解放军第四军医大学硕士学位论文
吞噬细胞免疫活性的调控，是机体抗肿瘤免疫的一个重要组成部分。
本实验采用 AgNORS图像分析定量测定方法、T淋巴细胞亚群检测试剂
盒（单克隆抗体直接免疫荧光染色法）、红细胞花环试验对86例恶性肿瘤患
者和42例正常对照组分别测定其外周血T淋巴细胞核仁银染区面积／细胞核
面积比值（IS％）、T 淋巴细胞亚群、红细胞免疫 C3b 受体花环率
（RBC（力）与红细胞循环免疫复合物花环率（RBCJCR卜结果显示：门）
肿瘤组 1＊％值减低，与 42例正常对照组比较差异十分显著少0刀 1）；u）肝
细胞癌组 L S％值的变化与临床分期密切相关，与病理分型无关；（3）肝细胞
癌组ISO／O值变化与肝硬化组，正常组有显著差异 V＜001），肝硬化组与正
常组之间有显著差异（＜0刀5）；u）肝细胞癌组1S％指标变化与淋巳结是否
有转移密切相关；（5）肿瘤组CD3”、CD4”、CD4－／CDS”比值减低，与正常对
照组比较差异十分显著（P叼01人＊仇－值与正常对照组比较无显著意义什
＞00s）；u）肿瘤组RBOQ＊g明显降低，RBC1＊R明显增高，与正常组比
较有显著差异（P＜001h（7）肿瘤组二＊％值与＊o丫0／比值呈正相关
（rro刀3民P＜0 01）；二＊％值与Q＊RR呈正相关（r＝0 503，P＜ o刀1）。
恶性肿瘤患者 IS％值减低，表明机体淋巴细胞增殖活性明显减低，I．S％
值是反映细胞增殖的敏感指标。恶性肿瘤患者CD。－／CD。－比值减低，表明T
细胞介导的细胞免疫功能降低和平衡失调，RBC－C力M降低，RBC－ICR增
高，提示RBC－C巾受体沾性减低及机体清除IC的能力减低。上述作用的结
乏 果是导致整个机体免疫系统功能降低，削弱了抗肿瘤能力。
本研究提示：图像分析法检测肿瘤患者外周血T淋巴细胞中核仁组成区
相关嗜银蛋白强度与机体免疫功能状态密切相关，对肿瘤患者的临床诊断，
治疗监测和判断预后提供一个新的简便可靠的方怯，在临床应用方面有重要
意义。
The pathogenesis of tumour is associated intimately with body s immune state.In fact, in normal body, 107-109 cells are undergoing mutation everyday. Tumour cells with malignant epitope are produced but usually tumour doesri t appear. An important reason is that the body s immunosurreillance can eliminate these cells. When the increase of tumour cells surpass the limit of immunosurreillance, tumour will be formed.
Bing responsible for transcriptional actively, the argyrophilic nuclolar regions (Ag-NORs) are acidic nonhistone proteins and are identified by a one-step silver staining technique. The argyrophilia appears to reflect rDNA transcription activity, which has been correlated with proliferative activity of cells. There are many studies about Ag-NORs on tumours, indicated that was correlation with
PCNA, Ki-67; benign and malignant tumour; potentially malignant lesions;grade of tumour.
T lymphocytes are the major immuno-active cells, which participate in the immunoreaction against tumour. Not only do they play a great role in cell immunity, but also do they play role in immunoregulation. The regulation is realized by its subgroup cells.
On the surface of red cell, there are many substances participating in the immunoreaction, such as CR^ CRs> CDs8> CD.s9> DAF^ SOD enzyme,etc. They all take part in the complex regulation of immunonetwork. Red cells have the function of immune adherence. They can adhere, transport and eliminate immune complex in circulation. They play role in the regulation of immune activity of T, B> NIC-. LAK cells and phagocytes, too. So red cells are an important part in the body' s immunity against tumour.
Nucleolar organizer regions and T lymphocyte subgroups (including CD3 , CD4*> CDg")of peripheral blood and erythrocyte immune function (including RBC-CsbRR and RBC-ICR) were measured in 86 patients with malignant tumours (such as liver,osteoma ,esophagus).The methods of Ag-NORs KL image-analysis,direct immunoflurescent staining technique with flow cytometry(FCM) and immune adherence rosette were used in the study.The results showed that:(l)The value of I.S% was lower in cancer patients than those of 42 healthy control subjects (PO.01). (2) The variation of I.S% ratio was significantly relative with clinical stages and was not relative with pathological classification in liver. (3) The value of I.S% was lower in liver patients than those of 22 cirrhosis and 42 healthy control subjects (P<0.01) and was stitistical different between cirrhosis and healthy control (P<0.05). (4) The value of I.S% was significantly relative with tumour transformation.^) The values of CDs*, CD4", CD47CDs~
were lower in cancer patients than those of 42 healthy control subjects (P<0.01) and the value of CDg+ was not statistical different between two groups CP>0.05).The values of RBC-C3bRR was lower and RBC-ICR was higher in the group of cancer patients as compared with the group of 42 healthy control subjects (P<0.01). (7)The value of I.S% significantly correlated with the CD4 /CD8~ ratio in the cancer groupe (r=0.713, PO.01) and the value of I.S% significantly correlated with the value of RBC-C3bRR in cancer groupe(r= 0.503, P<0.01).
The decreased I.S% ratio indicated that the cell proliferation was depressed. The I.S% ratio was a sensitive index of reflecting cell proliferation.The decreased CD4+/CDg^ ratio indicated by T lymphocyte was depresed and balance was interrupted. The decreased value of RBC-C3bRR and increased RBC-ICR demonstrated that RBC-Csb receptor activity and 1C elimination was significantly depressed in the patients with malignant tumours. The above effects result in the whole immune system function and the anti-tumour-immune activity depressed.
The Index I.S% significantly correlated with the immune function of body. The detected of I.S% ratio has important significance in clinical diagnosis and threapy evalueated and prognosis assessment of malignat tumourand a simple and exact methed.
引文
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46．陈万军，崔正言等。间接免疫荧光法与间接SPA法检测人外周血T细胞亚群的比较。上海免疫学杂志，1988；8(5)：389～391
47．陈善华，封江南，孙可芳，于箭，吴晓波，景岗，史良如。检测淋巴细胞亚群的花环试验的试制研究。免疫学杂志，1988：4(1)：49～52
48. Kammer GM, Stein RL, et al.T lymphocyte immune dysfunctions in systemic lupus erythematosus. J Labo Clin Med, 1990; 115:273～282
49．赵彩彦，刘金星，甄真等。肝硬化和肝细胞癌患者血清IL-2，SIL-2，IL-6，T细胞亚群的变化及相互关系。中国实用内科杂志，1999；1：22～24
50．赵风芹，宋玉明，刘晶，高申，王翠华。肺癌患者T淋巴细胞亚群NK细胞活性检测的临床监控。白求恩医科大学学报，1999；4：414～415
51．彭光洁，郭艺杰，曹学兵，胡群。急性白血病患儿血清肿瘤坏死因子-α和T淋巴细胞亚群的探讨。临床儿科杂志，1997；6；376～377
52．裘宇容。流式细胞仪对4种癌症病人T淋巴细胞亚群的检验。第一军医大学学报，1998；2：151～153
53．黄超，吴伟健。结肠癌患者T细胞亚群定量测定及其临床意义。广西医学，1997；19(5)：756～758
54．程凯灵，吴凯南，汪斌。癌症患者外周红细胞C_3b受体与CD_4/CD_8相关性探讨。中国肿瘤临床，1997；24(4)：269～271
55. Nelson JR.The immune-adherence phenomenon:A hypothetical role of erythrocytes in defence against bacteria and viruses. Pric Royeoe Med, 1956;49:55～58
56. Siegel I,The red cell immune system. Lancet, 1981;11 (8246): 556～559
57. Fearon DT. Identification of the membrane glycoprotein that is the C_3b receptor of the human erythrocyte polymorphnuclear leukocyte B lymphocyte and monocyte. J Exp Med, 1980; 152:20～24
58. Wilson JG, Woug WW, Schur PH, et al. Mode of inheritanced CR eased Nelson DS.Immune adherence ADV Immunol, 1963;3:131～133
59. Medof ME,Iida K, Mold C,et al. Unique role of the complement receptor C_1R in the degradation of C_3b assoiated with IC.J Exp Med, 1982;156:1739～1743
60．郭峰。红细胞免疫研究发展趋势。上海免疫学杂志，1995；15(4)：253～255
61. Comacoff JB,Hebert LA, Smmad WL,et al. Primate erythrocyte immune-complex-clearing mechanism. J Clin Invest, 1983;71:236～240
62. Schifferli JA, Walport MI.The clearance of tetanus tosoid/antitetanustxid IC from the circulation of humans. J Immunol, 1988; 140:899～904
63．崔瑞，郭峰。红细胞对巨噬细胞吞噬功能影响的观察。第二军医大学学报，1987；8(3)：207～209
64．项一萍。红细胞的免疫功能。国外医学输血液分册，1985；8(4)：206～208
65．李海旺，马晓慧，刘秀红。红细胞免疫检测技术在老年病的临床应用研究。中国免疫学杂志，1993；9(6)：374～375
66．刘景田，张浩。肿瘤红细胞免疫研究的新进展。陕西医学检验杂志，1993；8(4)251～256
67．崔英，郭峰，张其等。卵巢肿瘤患者红细胞免疫功能及超氧化物歧化酶的初步研究。第二军医大学学报，1995；16(6)：558～560
68．郭峰。红细胞增强淋巴细胞与粒细胞免疫粘附肿瘤细胞作用。第二军医大学学报，1989；10(6)：558～560
69. Virlla G,Rugeles MT, Hyman B,et al. The interaction of CD_2 with, it' s LFA-3 ligand espressed by autologous erythrocytes results inen hancement of B cell responses cellular. Immunology, 1988;116:308～319
70．郭峰，黄盛东，赫丽等。红细胞在肿瘤免疫反应中的作用。中华微生物学和免疫学杂志，1995；15(5)：183～187
71．郭峰。红细胞对肿瘤细胞有亲和性的发现。第二军医大学学报，1986；7(6)：426～430
72．郭峰，邬伟秀，蔡敏琪等。血液肺病患者红细胞免疫功能测定及其影响因素的研究。中华血液学杂志，1996；17(4)：209～211
73．李箭，施治，韦石松等。肿瘤患者血清对红细胞粘附作用的影响。上海免疫学杂志，1990；10：201～204
74．孙彦，刘忠强等。喉癌患者细胞免疫功能的检测。中华耳鼻喉科杂志，1995；30(6)：351～353
75. Gerdes J Schwab U, Lemke H, et al.production of A mouse monocolonal antibody reactive with a human nulear antigen associate with cell proliferotion. Int J Cancer, 1983;31:13～20
76. Kuwano H, Sumiyosh K,Nozoe T, Yasuda M, Wtanabe M, Sugimachi K.The
pronostic significance of the cytophtometric DNA content and its relation with argyrophilic nucleolar organizer regions(Ag-NOR) and proliferating cell nuclear antigen(PCNA) in oesophageal cancer.Eur J Surg Oncol,1995;21(4) :368-373
77． Okada H, Murai M,Yanmamoto H.Malignant fibrous histiocytoma: a case study of proliferative activity,immuohistochemistry and electron microscopy.J Oral Maxillofac Surg, 1994;52:1193-1197
78 De rosa I,Staibano S, Lo Muzio L,Delfmo M, Lucariello A,Coppola A,De rosa G,Scully C. Potentially malignant and malignant lesions of the lip role of silver staining nucleolar organizer regions,proliferating cell nuclear antigen, p53,and c-myc in differentiation and prognosis. J Oral Pathol Med,1999;28(6) :252-258
79． Tannapfel A,Geissler F,Kockerling F,Katalinic A,Hauss J,Wittekind C.Apoptosis and proliferation in relation to histopathological variables and prgnosis in hepatocellular carcinoma.J Pathol 1999; 187(4) :439-445
80． Zaitoun AM,Ebbs SR. Quantitative cytological assessment of Ki-67 and Ag-NORs using computer-digitized image analysis of four clinicopathological breast lesions,Cytopathology 2000; 11 (4) :243-253
81． Shi SR, KEY ME,Kalra KL. Antigen retrieval in formalin-fixed,paraffin-embedded tissues: An enhancement method for immunohistochemical staining based on microwave oven heating of tissus sections.J histochemistry and cytochemistry, 1991;6:41-48
82． Derezini M, Romagnoli T, Mingazzini P, Marinozzi V. Interphasic nucleolar organizer region distribution as a dignostic parameter to differentiate benign from malignant epithelial tumours of human intestine. Virchows Arch Bcell Pathol Incl Mol Pathol,1988;54(6) :334-340
83． Egan MJ,Crocker !.Nucleolar organizer regions in cutaneous tumours.J
Pathol,1988,154(3) :247-253
84． Ghazizadeh M, Sasaki Y,Oguro T,et al. Silver staining of nucleolar organizer in prostatic lesion.Histopath, 1991; 19:369-372
85． Lipponen PK,Eskelinen MJ,Nordling S. Nucleolar organizer regions as predictors in transitional cell bladder cancer. Br J Cancer, 1991,64:1139-1144
86． Lipponen PK.image analysis of Ag-NOR proteins in transitional cell bladder cancer.J Pathol, 1993; 171:279-283
87． Deckert M,Kubar Jbernar A.Functional cooperation between CD58 and CD59 in adhesion of T cell.J Immunol,1992;148:672-677
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34．苏燎原，徐映东，耿勇志等。淋巴细胞与肿瘤免疫的初探。免疫学杂志，1992；8(4)：251～252
35. Sercanz E,Erzych U.The distinctive specificity of antien-specific suppressor T cells. Immunol Today, 1991;12(4): 111～118
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45. Songsivilai S,Lachmann P, et al.Bispecific antibody:a tool for diagnosis and treatment of disease. Clin Exp Immunol, 1990;79:315～321
46．陈万军，崔正言等。间接免疫荧光法与间接SPA法检测人外周血T细胞亚群的比较。上海免疫学杂志，1988；8(5)：389～391
47．陈善华，封江南，孙可芳，于箭，吴晓波，景岗，史良如。检测淋巴细胞亚群的花环试验的试制研究。免疫学杂志，1988：4(1)：49～52
48. Kammer GM, Stein RL, et al.T lymphocyte immune dysfunctions in systemic lupus erythematosus. J Labo Clin Med, 1990; 115:273～282
49．赵彩彦，刘金星，甄真等。肝硬化和肝细胞癌患者血清IL-2，SIL-2，IL-6，T细胞亚群的变化及相互关系。中国实用内科杂志，1999；1：22～24
50．赵风芹，宋玉明，刘晶，高申，王翠华。肺癌患者T淋巴细胞亚群NK细胞活性检测的临床监控。白求恩医科大学学报，1999；4：414～415
51．彭光洁，郭艺杰，曹学兵，胡群。急性白血病患儿血清肿瘤坏死因子-α和T淋巴细胞亚群的探讨。临床儿科杂志，1997；6；376～377
52．裘宇容。流式细胞仪对4种癌症病人T淋巴细胞亚群的检验。第一军医大学学报，1998；2：151～153
53．黄超，吴伟健。结肠癌患者T细胞亚群定量测定及其临床意义。广西医学，1997；19(5)：756～758
54．程凯灵，吴凯南，汪斌。癌症患者外周红细胞C_3b受体与CD_4/CD_8相关性探讨。中国肿瘤临床，1997；24(4)：269～271
55. Nelson JR.The immune-adherence phenomenon:A hypothetical role of erythrocytes in defence against bacteria and viruses. Pric Royeoe Med, 1956;49:55～58
56. Siegel I,The red cell immune system. Lancet, 1981;11 (8246): 556～559
57. Fearon DT. Identification of the membrane glycoprotein that is the C_3b receptor of the human erythrocyte polymorphnuclear leukocyte B lymphocyte and monocyte. J Exp Med, 1980; 152:20～24
58. Wilson JG, Woug WW, Schur PH, et al. Mode of inheritanced CR eased Nelson DS.Immune adherence ADV Immunol, 1963;3:131～133
59. Medof ME,Iida K, Mold C,et al. Unique role of the complement receptor C_1R in the degradation of C_3b assoiated with IC.J Exp Med, 1982;156:1739～1743
60．郭峰。红细胞免疫研究发展趋势。上海免疫学杂志，1995；15(4)：253～255
61. Comacoff JB,Hebert LA, Smmad WL,et al. Primate erythrocyte immune-complex-clearing mechanism. J Clin Invest, 1983;71:236～240
62. Schifferli JA, Walport MI.The clearance of tetanus tosoid/antitetanustxid IC from the circulation of humans. J Immunol, 1988; 140:899～904
63．崔瑞，郭峰。红细胞对巨噬细胞吞噬功能影响的观察。第二军医大学学报，1987；8(3)：207～209
64．项一萍。红细胞的免疫功能。国外医学输血液分册，1985；8(4)：206～208
65．李海旺，马晓慧，刘秀红。红细胞免疫检测技术在老年病的临床应用研究。中国免疫学杂志，1993；9(6)：374～375
66．刘景田，张浩。肿瘤红细胞免疫研究的新进展。陕西医学检验杂志，1993；8(4)251～256
67．崔英，郭峰，张其等。卵巢肿瘤患者红细胞免疫功能及超氧化物歧化酶的初步研究。第二军医大学学报，1995；16(6)：558～560
68．郭峰。红细胞增强淋巴细胞与粒细胞免疫粘附肿瘤细胞作用。第二军医大学学报，1989；10(6)：558～560
69. Virlla G,Rugeles MT, Hyman B,et al. The interaction of CD_2 with, it' s LFA-3 ligand espressed by autologous erythrocytes results inen hancement of B cell responses cellular. Immunology, 1988;116:308～319
70．郭峰，黄盛东，赫丽等。红细胞在肿瘤免疫反应中的作用。中华微生物学和免疫学杂志，1995；15(5)：183～187
71．郭峰。红细胞对肿瘤细胞有亲和性的发现。第二军医大学学报，1986；7(6)：426～430
72．郭峰，邬伟秀，蔡敏琪等。血液肺病患者红细胞免疫功能测定及其影响因素的研究。中华血液学杂志，1996；17(4)：209～211
73．李箭，施治，韦石松等。肿瘤患者血清对红细胞粘附作用的影响。上海免疫学杂志，1990；10：201～204
74．孙彦，刘忠强等。喉癌患者细胞免疫功能的检测。中华耳鼻喉科杂志，1995；30(6)：351～353
75. Gerdes J Schwab U, Lemke H, et al.production of A mouse monocolonal antibody reactive with a human nulear antigen associate with cell proliferotion. Int J Cancer, 1983;31:13～20
76. Kuwano H, Sumiyosh K,Nozoe T, Yasuda M, Wtanabe M, Sugimachi K.The
pronostic significance of the cytophtometric DNA content and its relation with argyrophilic nucleolar organizer regions(Ag-NOR) and proliferating cell nuclear antigen(PCNA) in oesophageal cancer.Eur J Surg Oncol,1995;21(4) :368-373
77． Okada H, Murai M,Yanmamoto H.Malignant fibrous histiocytoma: a case study of proliferative activity,immuohistochemistry and electron microscopy.J Oral Maxillofac Surg, 1994;52:1193-1197
78 De rosa I,Staibano S, Lo Muzio L,Delfmo M, Lucariello A,Coppola A,De rosa G,Scully C. Potentially malignant and malignant lesions of the lip role of silver staining nucleolar organizer regions,proliferating cell nuclear antigen, p53,and c-myc in differentiation and prognosis. J Oral Pathol Med,1999;28(6) :252-258
79． Tannapfel A,Geissler F,Kockerling F,Katalinic A,Hauss J,Wittekind C.Apoptosis and proliferation in relation to histopathological variables and prgnosis in hepatocellular carcinoma.J Pathol 1999; 187(4) :439-445
80． Zaitoun AM,Ebbs SR. Quantitative cytological assessment of Ki-67 and Ag-NORs using computer-digitized image analysis of four clinicopathological breast lesions,Cytopathology 2000; 11 (4) :243-253
81． Shi SR, KEY ME,Kalra KL. Antigen retrieval in formalin-fixed,paraffin-embedded tissues: An enhancement method for immunohistochemical staining based on microwave oven heating of tissus sections.J histochemistry and cytochemistry, 1991;6:41-48
82． Derezini M, Romagnoli T, Mingazzini P, Marinozzi V. Interphasic nucleolar organizer region distribution as a dignostic parameter to differentiate benign from malignant epithelial tumours of human intestine. Virchows Arch Bcell Pathol Incl Mol Pathol,1988;54(6) :334-340
83． Egan MJ,Crocker !.Nucleolar organizer regions in cutaneous tumours.J
Pathol,1988,154(3) :247-253
84． Ghazizadeh M, Sasaki Y,Oguro T,et al. Silver staining of nucleolar organizer in prostatic lesion.Histopath, 1991; 19:369-372
85． Lipponen PK,Eskelinen MJ,Nordling S. Nucleolar organizer regions as predictors in transitional cell bladder cancer. Br J Cancer, 1991,64:1139-1144
86． Lipponen PK.image analysis of Ag-NOR proteins in transitional cell bladder cancer.J Pathol, 1993; 171:279-283
87． Deckert M,Kubar Jbernar A.Functional cooperation between CD58 and CD59 in adhesion of T cell.J Immunol,1992;148:672-677