宫颈病变细胞核AgNOR颗粒形态定量分析

为探讨不同程度宫颈病变的细胞核、ＡｇＮＯＲ的变化规律。利用自动图像分析技术分别对２８例宫颈癌，５５例宫颈非典型增生和１５例正常宫颈组织细胞核及ＡｇＮＯＲ颗粒形态定量参数进行测定。结果显示：除正常宫颈与轻度非典型增生宫颈组外，宫颈中度、重度非典型增生与宫颈鳞癌三组之间其平均细胞核面积、每个核ＡｇＮＯＲ颗粒数、平均颗粒面积、颗粒直径以及ＡｇＮＯＲ颗粒面积／细胞核面积之比值逐渐增加，而ＡｇＮＯＲ颗粒形状因子逐渐减小，各组之间存在着统计学差异。ＡｇＮＯＲ颗粒直径变化不显著。认为在宫颈组织发生癌变过程中，细胞核及核内ＡｇＮＯＲ颗粒大小、形态和数量均发生改变。提示细胞核及核内ＡｇＮＯＲ定量参数有可能作为估计肿瘤细胞恶性潜能的指标     

AgNOR的图像分析在判断子宫内膜癌预后方面的研究

应用图像分析技术对１０３例子宫内膜癌癌细胞中ＡｇＮＯＲ进行了定量研究。结果表明，低分化、肌层浸润＞１／２者的癌细胞核面积、每核ＡｇＮＯＲ颗粒总面积、颗粒数及颗粒平均面积明显大于高分化及无肌层浸润者。Ⅱ、Ⅳ期、复发、死亡患者的癌细胞的核面积及ＡｇＮＯＲ颗粒平均面积较Ⅰ期、无复发、存活者明显增高。ＡｇＮＯＲ的颗粒平均面积多在１．２０μｍ×１．２０μｍ以上，并且ＡｇＮＯＲ颗粒分布多呈聚集型；而预后较好的患者，ＡｇＮＯＲ颗粒平均面积多在亚．１５μｍ×１．１５μｍ左右，颗粒多呈弥散分布。结果提示，细胞内ＡｇＮＯＲ的含量与子宫内膜癌的生物学行为及预后有关，图像分析进行ＡｇＮＯＲ的定量研究较传统的光镜下计数ＡｇＮＯＲ颗粒数目更为客观、准确。     

胃癌和胃粘膜异型增生核仁组织区嗜银蛋白定量和形态研究

目的探讨核仁组织区嗜银蛋白（ＡｇＮＯＲ）染色技术在胃癌、癌前病变、慢性胃炎中鉴别诊断价值和意义。方法应用ＡｇＮＯＲ染色技术，观察１３２例胃良恶性病变和癌前病变细胞核中ＡｇＮＯＲ颗粒含量和形态。结果胃癌、胃粘膜异型增生、慢性胃炎三组间细胞核内ＡｇＮＯＲ颗粒均数差异非常显著（Ｐ＜０．０１），正常胃壁粘膜与慢性胃炎ＡｇＮＯＲ颗粒均数无明显差异（Ｐ＞０．０５）。胃癌和胃粘膜异型增生ＡｇＮＯＲ颗粒形态以弥散型为主，而慢性胃炎和正常胃粘膜以核仁型为主。结论细胞核内ＡｇＮＯＲ颗粒含量和分型对于区别胃良恶性及癌前病变有重要参考价值。     

AgNORs检验对胃粘膜良,恶性病变的诊断价值

为探讨ＡｇＮＯＲｓ变化对胃粘膜良，恶性病变的诊断价值，应用银染技术对５０例胃粘膜良，恶性病变进行ＡｇＮＯＲｓ计数，其中正常胃粘膜４例，萎缩性胃炎５例，异型增生９例，胃癌３２例结果。正常胃粘膜，萎缩性胃炎和异型增生细胞核内ＡｇＮＯＲｓ颗粒均数，分别与胃癌各型癌细胞核内均数比较均有显著性差异     

睾丸畸胎瘤核仁组成区嗜银蛋白图像分析

为进一步探讨睾丸良恶性畸胎瘤的鉴别方法，本文应用银染和图像分析技术，对睾丸正常组织、畸胎瘤、畸胎癌细胞进行了研究，结果发现：正常睾丸及畸胎瘤的ＡｇＮＯＲｓ颗粒呈圆形、规则、边界清楚、大小均匀，位于胞核偏中或边缘部位；畸胎癌的颗粒呈圆或卵圆形、欠规则、边界欠清、大小较不一致，偏中、散在或成簇分布于核内。畸胎癌的ＡｇＮＯＲｓ数目、面积、大颗粒数、平均面积、圆度比正常及畸胎瘤要大（Ｐ＜０．０１），前３项参数在畸胎瘤比正常组大（Ｐ＜０．０１）。故我们认为ＡｇＮＯＲｓ图像分析对睾丸良恶性畸胎瘤的鉴别具有意义，可为临床提供一项较为客观的指标。     

恶性淋巴瘤AgNORs定量分析的临床意义

采用核仁组成区相关嗜银蛋白（ＡｇＮＯＲｓ）染色技术，对７１例淋巴结良、恶性病变的组织石蜡切片进行ＡｇＮＯＲｓ定量研究。结果显示：３２例淋巴结良性病变平均每个细胞核内ＡｇＮＯＲｓ计数为１．７９±０．５９，３１例恶性淋巴瘤平均每个细胞核内ＡｇＮＯＲｓ计数为６．６２±３．０２，８例淋巴结内低分化转移癌平均每个细胞核内ＡｇＮＯＲｓ计数高达１３．７７±５．２４，表明良、恶性病变ＡｇＮＯＲｓ计数差别十分显著（Ｐ＜０．０１）。２０的非何杰金淋巴瘤（ＮＨＬ）中，低度恶性组ＡｇＮＯＲｓ计数为３．７９±１．４６，而中、高度恶性组ＡｇＮＯＲｓ计数为平均每个细胞核内ＡｇＮＯＲｓ８．２８±２．５８，两组比较差别有显著意义（Ｐ＜０．０１），显示低分化的恶性肿瘤，具有更加恶性的生物学行为。     

大肠癌细胞DNA、AgNORs定量研究──两者相关性及预后意义的探讨

应用ＡｇＮＯＲｓ染色技术，对６３例大肠癌的ＡｇＮＯＲｓ颗粒进行了计数研究。其中３８例同时进行肿瘤细胞核ＤＮＡ含量静态测定。结果表明，大肠癌细胞ＡｇＮＯＲｓ数量与其ＤＮＡ含量之间有明显相关性（Ｐ＜０．０ｌ）。ＤＮＡ含量和ＡｇＮＯＲｓ计数愈低，肿瘤分化愈好，患者预后愈好；反之，ＤＮＡ含量和Ａｇ－ＮＯＲｓ计数愈高、肝瘤分化愈差，患者预后愈差。表明ＡｇＮＯＲｓ计数，ＤＮＡ含量检测均可作为大肠癌分级、判断预后的参考指标。     

肝脏肿块针吸组织学和细胞学诊断及AgNOR分析

对４２例临床病理资料齐全的肝脏肿块针吸切片、涂片和涂片ＡｇＮＯＲ颗粒进行分析。切片为正常肝细胞４例、肝细胞异型７例、恶性肿瘤３１例。ＨＥ涂片与切片诊断符合率，在良性为１００％（１１／１１）例，恶性为８７％（２７／３１例）。３组涂片ＡｇＮＯＲ颗粒计数各为１.６４±０.７４、４.５２±１.２０和８.５３±１.８７，３组间差异非常显著Ｐ＜０.０１。以７个颗粒做良恶性分界，ＡｇＮＯＲ校正涂片诊断后，恶性符合率可达９７％（３０／３１例）。提议使用ＨＥ涂片定位脱色复染ＡｇＮＯＲ，严格选定被计数的细胞，对涂片误诊原因作了分析。     

氧自由基对大肠癌细胞rRNA基因活性和DNA含量的影响

目的探讨氧自由基对癌细胞核酸和基因活性的影响。方法采用计算机图像分析法测定叔丁基脂氢过氧化物（ｔｂｏｏＨ）对直肠腺癌ＨＲ８３４８细胞系间期细胞核及中期染色体上核仁组织区嗜银蛋白（ＡｇＮＯＲｓ）的作用，同时测定ＤＮＡ含量和细胞毒效应。结果１０－６ｍｏｌ／Ｌ的ｔｂｏｏＨ能使体外生长的大肠癌细胞出现细胞毒作用；ＡｇＮＯＲｓ颗粒在ｔｂｏｏＨ作用早期以聚集型为主，颗粒面积增大，而晚期颗粒分散，数量明显减少。早期ＡｇＮＯＲｓ颗粒增大并非是染色体上ｒＲＮＡ基因活性位点的增多，而是与间期细胞核内ＡｇＮＯＲｓ的聚集状态有关；晚期间期细胞内ＡｇＮＯＲｓ减少则是由ｔｂｏｏＨ引起染色体上ｒＲＮＡ基因位点减少所致。ｔｂｏｏＨ还可引起癌细胞ＤＮＡ含量减少。结论细胞基因活性或ＤＮＡ改变是自由基介导细胞损伤的重要机制。     

AgNORs检测对胃粘膜良、恶性病变的诊断价值

为探讨ＡｇＮＯＲｓ变化对胃粘膜良、恶性病变的诊断价值，应用银染技术对５０例胃粘膜良、恶性病变进行ＡｇＮＯＲｓ计数，其中正常胃粘膜４例，萎缩性胃炎５例，异型增生９例，胃癌３２例（高分化型腺癌８例，低分化型腺癌１８例，未分化癌６例）。结果：正常胃粘膜、萎缩性胃炎和异型增生细胞核内ＡｇＮＯＲｓ颗粒均数，分别与胃癌各型癌细胞核内均数比较均有显著性差异（Ｐ＜０．０１）。可以认为这种方法有助于区别胃粘膜良、恶性病变及其预后的判断。     

Alterations of the nucleolar organizer regions during 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in rats.

The number of silver-stained nucleolar proteins (AgNORs) was enumerated in preneoplastic and neoplastic rat tongue lesions induced by 4-nitroquinoline 1-oxide (4-NQO). Male ACI/N rats were given 0 to 10 p.p.m. 4-NQO for 12, 20 or 36 weeks to induce hyperplasia, dysplasia and neoplasm in tongue. The mean numbers of AgNORs stained by a modified one-step silver colloid method in various epithelial lesions were as follows: normal squamous epithelium (n = 5), 1.52 +/- 0.03; non-lesional squamous epithelium (n = 5), 1.58 +/- 0.04; hyperplasia (n = 20), 1.84 +/- 0.15; dysplasia (n = 20), 2.32 +/- 0.12; papilloma (n = 6), 2.23 +/- 0.10; and squamous cell carcinoma (n = 4), 3.06 +/- 0.26. Thus, the mean number of AgNORs showed a stepwise increase from untreated and treated, histologically normal squamous epithelium through hyperplasia and dysplasia to squamous cell papilloma and carcinoma, although the value of severe dysplasia was between those of papilloma and carcinoma. These results indicate that the mean number of AgNORs may reflect the proliferative nature of tongue lesions, as suggested in carcinogenesis of other organs, and also suggest that severe dysplasia is a direct precursor lesion for squamous cell carcinoma of the tongue induced by 4-NQO.     

Are argyrophilic nucleolar organizer regions good prognostic indicators of survival of patients with esophageal cancer with lymph node metastasis?

Argyrophilic nucleolar organizer regions (AgNORs) were evaluated in 95 samples from primary esophageal squamous cell carcinomas and 75 samples from metastatic lymph nodes. The number of AgNORs per nucleus in primary tumors with positive nodes (n = 53, 6.1 ± 1.8) was greater than that in primary tumors with negative nodes (n = 42, 3.8 ± 1.1, P < 0.001). In 39 of 53 patients with positive nodes, the numbers of AgNORs per nucleus in metastatic lymph nodes were lower than those in primary tumors. The 5-year survival rate of these patients was 23.7%. However, the numbers of AgNORs per nucleus in metastatic lymph nodes were greater than those of primary tumors in 14 of 53 patients with positive nodes, and 11 of these 14 patients died from recurrence of cancer within 3 years after surgery. These observations suggest that the proliferative activity of cancer cells might be suppressed in the regional lymph nodes. However, cancer cells with higher proliferative activity in the regional lymph nodes than in the primary tumors might overcome immunological defenses and subsequent further metastasis might occur. © 1996 Wiley-Liss, Inc.     

Evaluation of the nucleolar organizer region associated proteins in exfoliative cytology of normal buccal mucosa. Effect of smoking

An evaluation of cellular alterations in the smoker's oral mucosal cells was performed. The Exfoliative Cytology technique was applied and the cytological smears stained with silver for the enumeration of Argyrophilic nucleolar organizer regions (AgNORs). Cytological smears were collected from two anatomic sites: floor of the mouth and tongue border, in both groups, smokers and non smokers, with a purpose of correlating the smoking habit to possible cellular alterations. The enumeration of the AgNORs showed that the average number of AgNORs is higher in smokers. There is a significant difference (P=0.0001) between smears from the floor of the mouth and from tongue border in the smoking group. In this study, no correlation between number of cigarettes, age and gender was found, but the results suggest that there might be a correlation between the smoking habit and an increased rate of cellular proliferation in the oral mucosal cells.     

Evaluation of argyrophilic nucleolar organizer regions in tongue squamous cell carcinomas

Using a silver staining technique, argyrophilic nucleolar organizer regions (AgNORs) were studied on the biopsy specimens taken from 67 tongue squamous cell carcinomas (SCCs), and the relations of the results of AgNORs to the clinicopathological findings, to the labelling indices of proliferating cell nuclear antigen (PCNA LI), and to the outcomes of the disease were investigated. The mean numbers of AgNORs per cell were 1.67 +/- 0.19 (n = 27) in the control squamous epithelia and 3.58 +/- 1.15 (n = 67) in the tongue SCCs, respectively, and a significant difference was found between the two groups (P < 0.0001). Concerning the clinicopathological findings, the mean numbers of AgNORs in the progressive cases (T3, T4), in the factor N-positive groups (N1, N2), and in the advanced stages (stages III, IV) were statistically higher than those in T1, T2, N0, and earlier stage (stages I, II) tumours, respectively. Similarly, a higher value of the AgNOR count was present in the histological grade III or diffuse invasive tumour. There was also a directly significant correlation between the AgNOR counts and the labelling indices of PCNA (r = 0.53, P < 0.0001). Concerning the outcome of the disease, the mean numbers of AgNORs were higher in the group with local recurrence or with lymph-node metastasis. A lower rate of 5 years' survival was found in the high value group (63.5%) of AgNORs compared with that of the low value group (86.0%), with a significant difference between the two groups (P < 0.05). The results suggest that AgNORs may reflect the degree of malignancy and cellular proliferation in tongue SCCs.     

The relation of argyrophilic proteins of nucleolar organizer regions (AgNORs) to the proportions of Ki-67 or DNA polymerase alpha-reacting cells in non-Hodgkin's lymphomas.

In order to examine the relationship between argyrophilic proteins of nucleolar organizer regions (AgNORs) and the proliferation activity of cells, we investigated lymph nodes obtained from 25 untreated non-Hodgkin's lymphoma (NHL) patients. Two monoclonal antibodies (MoAb) (Ki-67 antibody and anti-DNA polymerase alpha antibody) were used for evaluating cell proliferation activity. A linear relation between the mean number of agNORs per nucleus and the proportion of NHL cells reacting with Ki-67 MoAb was observed (r = 0.48, P less than 0.05). A similar relation between AgNORs and DNA polymerase alpha MoAb was also observed (r = 0.51, P less than 0.01). From these data, it was confirmed that AgNORs reflect the proliferation activity of NHL cells. We conclude that the AgNOR staining procedure is one of the simplest and most reliable methods for analyzing cell proliferation potential.     

图像分析技术测定AgNORs对儿童NHL预后的意义

用图像分析仪测定３３例儿童石蜡包埋组织的ＡｇＮＯＲｓ参数，探讨ＡｇＮＯＲｓ参数与儿童ＮＨＬ生存期有关系及临床工作分类与ＡｇＮＯＲｓ参数的关系。结果显示：１）生存与ＡｇＮＯＲｓ参数显著相关生存〈１年和〉１年，ＡｇＮＯＲｓ参数差异极显著。ＡｇＮＯＲｓ数量多，面积大，生存时间短，反之亦然。２）年龄，工作分类与ＡｇＮＯＲｓ显著     

肝癌细胞抗原致敏树突状细胞诱导的体外抗肿瘤作用

目的　探讨原发性肝癌患者外周血树突状细胞 (DC)经自体肝癌细胞抗原致敏后诱导的体外抗肿瘤作用。方法　对肝癌患者外周血采用密度梯度离心法分离 ,获得DC前体细胞 ,用重组人粒细胞 巨噬细胞集落刺激因子 (rhGM CSF)和重组人白细胞介素 4(rhIL 4)联合培养 ,诱导扩增DC。制备自体肝癌细胞抗原 ,体外脉冲DC ,检测DC诱导自体T细胞增殖能力及细胞毒性T细胞 (CTL )在体外对自体肝癌细胞的杀伤活性 ,并检测肿瘤抗原致敏DC分泌的IL 12水平。结果　经自体肝癌细胞抗原致敏的DC能分泌IL 12和诱导较强的自体T细胞增殖 ,且能诱导特异性CTL ,该CTL对自体肝癌细胞具有很强的杀伤活性 ,杀伤率显著高于DC、未经肝癌细胞抗原致敏的DC激活的CTL及T淋巴细胞的杀伤率 ,而对CT 2 6细胞、BEL 740 2细胞无明显的杀伤作用。结论　肝癌患者外周血DC经自体肝癌细胞抗原致敏后能诱导高效而特异的抗肝癌免疫 ,其机制可能与增强T细胞应答和诱导机体产生肿瘤特异CTL从而发挥特异性的抗肿瘤作用有关。     

肝癌射频治疗后的DSA征象分析

背景与目的:肝射频消融术(RFA)是肝癌综合治疗中的一项重要手段,CT常用于评估其治疗效果和肿瘤复发,但CT很难发现＜1 cm的复发灶,因此必要时行血管造影术(DSA)能发现隐匿的复发灶,是CT有益的补充手段.为此,本文研究肝癌RFA后的DSA征象,以提供影像学参考.方法:2007年1月至2008年4月对17例经临床、影像学及肝穿刺活检确诊为肝癌或转移性肝癌患者行RFA后进行DSA造影.结果:DSA显示射频治疗的肿瘤区多为圆形或类圆形无染色的低密度区;边缘区可见以下几种征象:局部染色、肝动脉门静脉瘘、边缘区出血、边缘复发和无异常造影征象.肝内异位复发灶造影表现同其原发肿瘤常见造影表现.本组9例造影发现原位边缘复发和(或)肝内异位复发灶.结论:发现和鉴别射频治疗区域的边缘征象是判断局部复发的关键.DSA在榆出＜1 cm的边缘结节复发和肝内复发灶方面优于CT.     

Prognostic value of AgNOR counting

The prognostic significance of the argyrophilic nucleolar organizer regions (AgNORs) in tumour pathology is still a matter of debate. A prospective study was performed in a series of renal cell carcinomas to clarify the prognostic value of AgNOR counting. Sections from 21 renal cell carcinomas were stained in 1990 with the method of Ploton. Black dots within the nucleus from 200 tumour cells were counted: the mean AgNOR count for the whole series was 6.13, the median 5.94 and the SD 1.78. Patients were then followed up for at least 6 years or to death: at the time of the survival analysis (June 1996), 13 patients were alive without evidence of recurrence or metastasis, 6 had died of the disease and 2 of myocardial infarction. All the patients with 5.94 AgNORs per cell or fewer were alive at 6-year follow-up, while only 60% of patients with more than 5.94 AgNORs per cell survived (p=0.01). In the multivariate analysis, only AgNOR count (p=0.015) retained an independent prognostic significance. With the limitation due to the small number of cases, this prospective study clearly indicates that AgNOR count has a significant prognostic role, at least in renal cell carcinoma.