大肠肿瘤P0CNA、AgNOR、DNA及核形态定量研究

目的：研究大肠上皮异型增生多指标多参数检测及其在大肠肿瘤的计算机自动化诊断中的应用价值。方法：本课题应用计算机图像分析技术对53例大肠癌、30例大肠腺瘤及10例正常肠粘膜进行DNA含量、核形态参数、AgNOR的计数、颗粒形态参数及PCNA等多指标多参数检测。结果：多数参数在各组间具有显著性差异（P＜0．01）,经判别筛选,DNA指数、AgNOR的平均颗粒数、阳性率、平均光密度、颗粒表面因子,PCNA的平均光密度、阳性率,核的形态因子、 核宽径及平均光密度等10个参数判别能力较大,为最佳判别参数。参数值大多随着正常、腺瘤及腺癌顺序递增,显示这些参数均能较好反映肿瘤的演进过程。PCNA大部分参数值在肿瘤的演进过程 中其高峰出现在腺瘤阶段,故其在大肠癌早期诊断中的价值需进一步探讨。结论：应用图像分析技术对大肠癌及癌前病变进行多指标多参数的计量分析,具有鉴别诊断有重要意义,为大肠肿瘤的自动化诊断提供客观依据。     

甲状腺滤泡性肿瘤DNA含量与AgNOR计数研究

目的：探讨DNA含量与AgNOR对甲状腺良恶性滤泡性肿瘤的鉴别诊断价值。方法：应用流式细胞术和胶体银染技术对9例正常甲状腺组织和36例滤泡性肿瘤（22例腺瘤和14例腺癌）进行DNA含量分析和AgNOR计数。结果：14例滤泡性癌中，11例为DNA异倍体，而22例滤泡性腺癌仅1例为DNA异倍体，且伴有不典型增生。AgNOR由正常甲状腺组织、滤泡性腺癌至滤泡性癌逐渐增加，相互之间有显著性差异（P＜0．05）；但各组间AgNOR计数有重叠，且DNA含量与AgNOR无明显相关性。结论：尽管AgNOR染色方法简便。经济，但在区别良恶性滤泡性肿瘤方面，DNA含量异常更具意义。     

大肠肿瘤多项生物标志定量检测及判别分析

目的:对大肠上皮异型增生多指标多参数进行检测,探讨其在大肠癌早期诊断中的应用价值。方法:应用计算机图像分析技术对大肠癌、不同增生程度的大肠腺瘤及正常肠粘膜进行DNA含量、核形态参数,AgNOR的计数、颗粒形态参数及PCNA、c-erbB-2、p53等多指标多参数检测,并应用数学统计方法建立判别函数。结果:经判别筛选,DNA指数,AgNOR阳性率、平均光密度、颗粒表面因子,c-erbB-2的平均光密度,p53的平均光密度,核的形状因子及平均光密度等8个参数判别能力较大,参数值大多随着正常肠粘膜→Ⅰ级不典型增生腺瘤→Ⅱ级不典型增生腺瘤→Ⅲ级不典型增生腺瘤→腺癌的顺序呈递增趋势,且组间差别有显著性(P<0.01),显示这些参数均能较好反映肿瘤的演进过程,由此建立的判别函数回代符合率达96.45%,结果优于单一指标建立的判别。PCNA在大肠癌早期诊断中的价值需进一步研究。结论:应用图像分析技术对大肠癌及癌前病变进行多指标多参数的计量分析,具有鉴别诊断的重要意义,为大肠癌的早期诊断提供客观依据。     

恶性纤维组织细胞瘤PCNA与AgNOR定量检测及其预后意义

目的：为探讨恶性纤维组织细胞瘤的增殖细胞核抗原（PCNA）及核仁组成区嗜银蛋白（AgNOR）定量检测与肿瘤恶性及预后的关系。方法：应用免疫组织化学染色LSAB法及AgNOR染色法46例资料完整有随结果的恶性纤维组织细胞瘤（MFH）手术切除标本进行染色以显示PCNA及AgNOR,彩色病理图像分析系统定量检测。结果：肿瘤细胞核PCNA过表达及AgNOR计数及患生存期呈负相关（rPCNA=-0.6248,rAgNOR=-0.5941,P<0.01);PCNA及AgNOR定量高值组较之定量低值组患生存期短（P＜0．01）;肿瘤组PCNA及AgNOR定量高于无转移组（P＜0．01）;肿瘤位置深在组PCNA及AgNOR定量高于肿瘤位置浅表组（P＜0．01）。结论：肿瘤细胞核PCNA及AgNOR定量与患生存期密切相关,与肿瘤转移及位置深浅等反映肿瘤恶性度的指标亦密切相关,提示肿瘤细胞核PCNA及AgNOR定量对于判断恶性纤维组织细胞瘤恶性度及预后的意义值得重视。     

子宫内膜增生与内膜癌增殖细胞核抗原表达及AgNOR计数关联研究

应用抗增殖细胞核抗原（PCNA）的单克隆抗体，以ABC免疫组织化学方法及银染色方法对112例子宫内膜标本进行测定，并与病理学诊断进行对比分析。结果表明：在各种类型的子宫内膜中，PCNA的表达及AgNOR计数依病变的进展明显递增。子宫内膜癌组PCNA表达及AgNOR计数明显高于正常子宫内膜组及子宫内膜增生组（增殖期例外），各组相互比较有显著差异（P＜0．01）。此种方法是测定子宫内膜增生细胞增殖分数的有效手段。对于判断其生物学行为及预后有重要意义，可做为临床判断良恶性子宫内膜增生的指标之一。     

甲状腺肿瘤细胞DNA含量及Cyclin D1与p27和Rb基因表达的定量检测

目的：探讨DNA含量(D1值)、细胞增殖活性(PI值)、Cyclin D1、p27及Rb基因表达在不同甲状腺肿瘤中的变化，为甲状腺癌的癌变机制及甲状腺肿瘤的鉴别诊断提供实验依据。方法：采用流式细胞术(FCM)和免疫荧光技术对甲状腺癌(40例)、结节性甲状腺肿(21例)、甲状腺腺瘤(16例)、正常甲状隙组织(10例)的DNA含量、细胞增殖活性及Cyclin D1、p27、Rb基因蛋白的表达进行定量检测结果：甲状腺癌组的D1值明显增高，异倍体细胞明显增多，异倍体率为80．0％，PI值明显增高Cyclin D1蛋白在甲状腺癌组表达显著增高，p27蛋白表达量在结节性甲状腺肿、甲状腺腺瘤和甲状腺腺癌组中依次减少，均明显低于正常甲状腺组织。结论：甲状腺癌发展过程中，DNA含量及异倍体率增加，细胞增殖活性增高Cyclin D1蛋白高表达，p27蛋白低表达，Cyclin D1、p27与甲状腺肿瘤的发生密切相关。     

喉癌细胞PCNA表达和DNA含量与临床预后的关系

 目的　探讨增殖细胞核抗原(PCNA)的表达和DNA含量对喉癌临床生物学特征及预后的关系。方法　用免疫组化SP方法和自动图像分析仪检测正常喉粘膜、喉鳞癌的PCNA和DNA指数(DI)，结合临床随访资料进行分析。结果　喉鳞癌PCNA的阳性表达率为100%，与正常喉组织有显著差异(P<0.01)，PCNA阳性表达强度和DNA指数随肿瘤分化程度的降低而相应增高(P<0.05)。两指标较高者生存时间较短，预后较差(P<0.05)。结论　PCNA的表达和DNA指数可为判断肿瘤的恶性程度及临床预后提供参考依据。     

恶性纤维组织细胞瘤PCNA与AgNOR定量检测及其预后意义

目的为探讨恶性纤维组织细胞瘤的增殖细胞核抗原（PCNA）及核仁组成区嗜银蛋白（AgNOR）定量检测与肿瘤恶性度及预后的关系。方法应用免疫组织化学杂色LSAB法及AgNOR染色法对46例资料完整有随访结果的恶性纤维组织细胞瘤（MFH）手术切除标本进行染色以显示PCNA及AgNOR,彩色病理图像分析系统定量检测。结果肿瘤细胞核PCNA过表达及AgNOR计数与患者生存期呈负相关（rPCNA＝－0．6248、rAgNOR＝－0．5941,P＜0.01）;PCNA及AgNOR定量高值组较之定量低值组患者生存期短（P＜0．01）;肿瘤有转移组PCNA及AgNOR定量高于无转移组（P＜0．01）;肿瘤位置深在组PCNA及AgNOR定量高于肿瘤位置浅表组（P＜0．01）。结论肿瘤细胞核PCNA及AgNOR定量与患者生存期密切相关,与肿瘤转移及位置深浅等反映肿瘤恶性度的指标亦密切相关,提示肿瘤细胞核PCNA及AgNOR定量对于判断恶性纤维组织细胞瘤恶性度及预后的意义值得重视。     

大肠肿瘤中P53和PCNA表达的定量研究

目的 研究大肠肿瘤中P53及PCNA定量表达及临床意义。方法 应用免疫组化和图像分析技术对40例腺瘤、80例腺癌及15例正常粘膜P53和PCNA的含量进行研究。结果 正常粘膜P53表达阴性,腺瘤和腺癌中P53蛋白表达量差异显著(P<0.05),PCNA在正常粘膜、腺瘤、腺癌中表达量依次递增,且差异显著(P<0.01),PCNA表达与大肠癌的分化有关,与其它临床病理因素无关(P>0.05)。结论 P53和PCNA表达在大肠肿瘤发生中起重要作用,检测P53和PCNA表达量的变化有利于大肠癌早期诊断和预后判断。     

结直肠癌及癌前病变中DNA倍体分析及其临床意义

目的:检测结直肠癌及癌前病变细胞内的DNA含量及DNA指数,探讨其辅助诊断和预测癌变危险度的意义.方法:应用流式细胞仪(FCM)对400例结直肠组织细胞的细胞核进行DNA倍体分析,其中正常结直肠黏膜组30例,增生性息肉组30例,绒毛状腺瘤组190例,结直肠癌组150例,检测其异倍体检出率和DI值.结果:结直肠癌组的DNA异倍体检出率显著高于正常直肠黏膜组3、增生性息肉组及绒毛状腺瘤组(P＜0.01),绒毛状腺瘤伴重度不典型增生组的DNA异倍体检出率显著高于正常直肠黏膜组3、增生性息肉组、无不典型增生的绒毛状腺瘤组及伴轻度、中度不典型增生的绒毛状腺瘤组(P＜0.01).随着结直肠癌的组织学分化程度降低,其DNA异倍体DI值呈增加趋势并有显著性差异(P＜0.01).结论:DNA含量和倍体的检测有助于了解肿瘤细胞的增殖情况及恶性程度,对诊断结直肠良恶性肿瘤、预测癌变危险度、判断预后均具有重要的参考价值.     

增殖细胞核抗原与AgNOR检测在前列腺癌临床中的意义

目的研究前列腺癌中增殖细胞核抗原（ＰＣＮＡ）表达及ＡｇＮＯＲ计数的意义。方法应用免疫组织化学方法和银染色技术检测前列腺癌和良性前列腺增生组织中ＰＣＮＡ表达和ＡｇＮＯＲ计数。结果ＰＣＮＡ增殖指数与ＡｇＮＯＲ计数在癌组织中均明显高于良性前列腺增生组织（Ｐ均＜０．００１）,且两者均与肿瘤组织学分级和预后有密切关系（Ｐ均＜０．０１和Ｐ均＜０．００１）;前列腺癌ＰＣＮＡ增殖指数与ＡｇＮＯＲ计数间存在非常显著的正相关（Ｐ＜０．０１）。结论ＰＣＮＡ增殖指数和ＡｇＮＯＲ计数结合分析,在鉴别前列腺良恶性病变和判断前列腺癌的恶性程度及预测患者预后方面具有十分重要的意义。     

肾癌细胞增殖活性与预后的关系

目的:为了探索肾癌增殖细胞的生物学特性与预后的关系.方法:应用免疫组化ABC法和流式细胞术DNA分析技术.结果:检测42例肾癌PCNA阳性率(LI)平均为12.4±5.1%.其中二倍体18例,LI值为2.41±3.14%;异倍体24例,LI值为6.80±4.29%.以LI均值为界分组,A组LI值≤12.4%,有23例,5年生存率为60.9%(14/23);B组LI值>12.4%,有19例,5年生存率为15.8%(3/19).两组差异非常显著(P<0.01).结论:提示PCNA的LI值可作为判断恶性度的客观指标,DNA直方图可评估预后.     

Proliferating cell nuclear antigen and argyrophilic nucleolar organizer regions in patients with thymic disease

We examined argyrophil nuclear organizer regions (AgNOR) and proliferating cell nuclear antigen (PCNA) in 41 patients with surgically-treated thymic disease. AgNOR count and PCNA labeling index (LI) in thymic carcinoma were significantly higher than those in thymoma and thymic hyperplasia. A positive correlation was observed between the PCNA LI and argyrophilic nucleolar organizer region (AgNOR) counts within all thymic disease (r=0.31, P=0.002). The PCNA LI in invasive thymoma was lower than that in non-invasive thymoma. In survival analysis, the cut-off values for the PCNA LI and AgNOR count were chosen to produce two categories with equal numbers of 26 thymoma patients. There were no significant difference in the survival rate between the lower and higher group patients in relation with AgNOR count and PCNA LI. We conclude that combining AgNOR and PCNA may discriminate the biological activity of thymic disease. These staining methods can be performed with ease and, applied in a clinical laboratory on a routine basis to help predict cytological malignancy of thymic disease.     

The estimation of proliferative activity by pcna and AgNORs in leukoplakia and squamous-cell carcinoma of the oral cavity

The proliferating activity of leukoplakia and squamous cell carcinoma (SCC) was estimated using proliferating cell nuclear antigen (PCNA) immunostaining and silver-binding argyrophilic nucleolar organizer region (AgNOR) staining. Twenty-eight leukoplakias and 15 SCCs of the oral cavity were used in this study. The mean+/-S.D. of PCNA LI and AgNOR counts were 7.7+/-6.0% and 2.43+/-0.68/nucleus in leukoplakias, and 22.3+/-11.6% and 4.77+/-1.49/nucleus in SCCs. Both of PCNA LI and AgNOR counts were significantly higher in SCCs than in leukoplakias. There was a significant linear correlation between PCNA LI and AgNOR counts (p=0.0022) in leukoplakias, but not in SCCs. In the series of leukoplakias, PCNA LI apparently increased in leukoplakias with dysplasia and malignant transformed cases. Our data suggest that PCNA LI and AgNOR counts are useful markers of proliferating activity, and PCNA LI might be a prognostic factor of leukoplakias.     

胃癌细胞DNA倍性,增殖活性与预后的关系

用液式细胞术（ＦＣＭ）测定７４倒胃癌细胞ＤＮＡ倍性，Ｓ期细胞比率（ＳＰＦ）和增殖指数（ＰＩ），以１８例正常胃粘膜作为正常对照。结果：胃癌组６６．２３％（４９／７４）检出异倍体，正常组均为二倍体，两者差异显著（Ｐ＜０．０１）．而胃癌组ＳＰＦ和ＰＩ也均显著高于对照组（Ｐ＜０．０１）．ＳＰＦ，ＰＩ与胃癌淋巴结转移、切端癌残留有关（Ｐ＜０．０１，Ｐ＜０．０５）．异倍体胃癌患者五年生存率显著低于二倍体胃癌（Ｐ＜０．０５）．年龄＞５５岁组的异倍体胃癌预后更差（Ｐ＜０．０５）．ＳＰＦ，ＰＩ与胃癌术后生存呈显著负相关（Ｐ＜０．０１）．ＦＣＭ胃癌细胞ＤＮＡ倍性、增殖活性检测有助于胃癌恶性度和预后的判断。     

Argyrophilic nucleolar organizer regions in breast carcinoma. Correlation with DNA flow cytometry, histopathology, and lymph node status.

Argyrophilic nucleolar organizer regions (AgNOR) have been correlated with proliferative activity of neoplasms. Increased AgNOR may reflect increased proliferative activity of cells or ploidy. To explore this hypothesis, 41 breast carcinomas were processed for AgNOR silver staining and DNA flow cytometry. AgNOR counts were expressed as mean AgNOR/nucleus and percentage of tumor cells with more than five AgNOR/nucleus. The first count was designated mean AgNOR or mAgNOR, and the second count was designated AgNOR proliferative index or pAgNOR. Using Mantel-Haensel statistical analysis, carcinomas that exhibited mAgNOR of 2.4 or more had a high likelihood of aneuploidy (P less than 0.0001), an S-phase fraction of more than 5.8% (P less than 0.003), or a diameter greater than 2 cm (P less than 0.007). In addition, tumors with pAgNOR of 8% or more showed a statistically significant correlation with aneuploidy (P less than 0.004), tumor grade (P less than 0.04), and a more significant one with high S-phase fraction (P less than 0.0001). No significant correlation was obtained between pAgNOR and tumor size or lymph node status. These data indicate that AgNOR quantitation reflects changes in DNA ploidy and cell proliferation. They also suggest that the mean AgNOR counts correlate best with the DNA mass or ploidy and that the frequency of cells with higher AgNOR count best reflects proliferative activity or S-phase fraction.     

DNA ploidy and PCNA index in pancreatic lesions producing hyperinsulinemic hypoglycemia

DNA nuclear content and PCNA index (proportion of PCNA reactive cells) have been studied by flow cytometry in eight pancreatic lesions producing hyperinsulinemic hypoglycemia to assess DNA ploidy and tumoral growth fraction. Adult nesidioblastosis had a diploid DNA index, two adenomas were near diploid, and four of the adenomas and the carcinoma were aneuploid. The median value of PCNA index (6.056% ± 6.76) was significantly correlated (p <0.05) with the mean tumor diameter (2.43 cm ± 1.96). Tumors with a PCNA index <6.056% showed a diameter less than the mean tumoral diameter and a benign clinical course. Tumors with PCNA index <6.056% generally displayed a diameter bigger than the mean tumoral diameter, being associated with lymph node metastases in one case. The authors conclude that nuclear DNA and PCNA index cytometric studies are useful parameters to assess the biological behaviour of pancreatic lesions producing hyperinsulinemic hypoglycemia.     

DNA含量及S期细胞比值和胃癌生物学特性的关系

研究ＤＮＡ含量及ＳＰＦ值与胃癌临床病理指标及ＰＣＮＡ表达的关系。方法：应用流式细胞术测定７２例胃癌新鲜组织及３１例良性胃粘膜病变新鲜组织的ＤＮＡ含量ＳＰＦ值,并采用免疫组织化学方法检测６６例胃癌石蜡切片的ＰＣＮＡＬＩ,并结合临床资料分析其临床意义。结果胃癌的ＤＩ（１．２１）及异倍体检出率（４４．４４％）,明显高于良性胃粘现变组（分别为１．０５和９．６８％）吕患者中异倍体组的ＳＰＦ值（２２．７４％）     

前列腺癌AgNOR计数与DNA含量测定的意义

作者应用银染色技术对３０例前列腺癌、２０例前列腺增生症组织中的核仁组成区相关嗜银蛋白（ＡｇＮＯＲ）进行检测，并采用图像分析技术测定其ＤＮＡ含量。结果显示，在前列腺癌组织中，ＡｇＮＯＲ计数与ＤＮＡ含量均明显高于前列腺增生症组织（Ｐ＜０．００１）；肿瘤分化越差，ＡｇＮＯＲ计数和ＤＮＡ含量越高，并且与患者预后相关（Ｐ＜０．００１）；ＡｇＮＯＲ计数与ＤＮＡ含量呈明显的正相关（Ｐ＜０．０１）。结果提示ＡｇＮＯＲ计数与ＤＮＡ含量可作为反映前列腺癌生物学行为的指标。