Argyrophilic nucleolar organizer regions in human adrenocortical neoplasms

Summary Argyrophilic nucleolar organizer regions (AgNOR) in human adrenocortical neoplasms, including five carcinomas and ten adenomas, were studied using a semi-automatic image analyzer. Both the number and total area of AgNOR per nucleus in the carcinomas were found to be statistically greater than in adenomas and control tissues. However, there were no statistically significant differences in total AgNOR area per nuclear area or in the mean area of individual AgNOR dots. The AgNOR of neoplastic and normal cells were of four morphological types: type 1 had a few dots at the periphery of the nucleus, type 2 a few dots at the center, type 3 a large round dot along with several small ones at the center, and type 4 numerous diffusely distributed polymorphic dots. Most type 3 and 4 cells were found in carcinoma cases. Type 1 cells decreased in proportion to the severity of biological malignancy. It follows from these findings that careful observation of AgNOR should facilitate the distinction of malignant from benign adrenocortical neoplasms.Abbreviation AgNOR argyrophilic nucleolar organizer region     

肺结核病人T淋巴细胞核仁区酸性非组蛋白表达活性的研究

目的　探讨结核病人T淋巴细胞核仁区酸性非组蛋白 (Ag-NORs)的数量,间接反映T淋巴细胞的功能,及其在结核病诊断及发病中的意义。方法 使用北京开隆公司提供的KL型免疫图像分析系统对41例正常人,77例肺结核病人和32例肿瘤病人的T淋巴细胞核仁区酸性非组蛋白表达活性进行对比,以核仁染色区面积与细胞核面积之比 (I.S%)定量反映Ag-NORs的多少。结果 正常人I.S%为 (7.20±0.95)%,肺结核病人为 (5.59±1.47)%;肿瘤病人为 (5.04±1.52)%,二者与正常人相比有显著性差异 (P＜0.001)。结核病人略高于肿瘤病人,他们之间无显著性差异。结论 结核病人酸性非组蛋白表达检测,可用于对结核病人免疫功能的检测,对估计病人的预后有临床指导意义。     

Argyrophilic nucleolar organizer region counts and proliferating cell nuclear antigen scores are two reliable indicators of survival in pharyngeal carcinoma

Summary The proliferative activity of pharyngeal carcinoma has been investigated by means of monoclonal antibody PC10 against proliferating cell nuclear antigen (PCNA/cyclin) and argyrophilic nucleolar organizer region (AgNOR) analysis in formalin-fixed, paraffin-embedded biopsies from 45 primary squamous and undifferentiated carcinomas, prior to therapy.The correlation between AgNOR counts and PCNA(PC10) scores was highly significant (r=0.73;P<0.0001) as determined by Pearson's correlation coefficient. Moreover, the univariate Kaplan-Meier survival analysis showed a significant correlation between 3- and 5-year survival rates and the mean AgNOR number per tumour cell (P=0.0003) or the percentage of PCNA(PC10)-positive cells (P=0.0001).Our results indicate that both AgNOR counts and PCNA(PC10) scores are reliable markers of the proliferative activity of pharyngeal carcinoma in small, routinely processed biopsies, in which they can allow simultaneous evaluation of the histology and tumour cell kinetics.Abbreviations AgNOR argyrophilic protein associated with nucleolar organizer regions - PCNA proliferating cell nuclear antigen This work was supported by grants from the Italian Ministero dell'Universitá e Ricerca Scientifica e Tecnologica (60%) and Lega Italiana per la lotta contro i tumori, Sezione di Novara.     

Demonstration of nucleolar organizer regions in intrahepatic bile duct carcinoma by the silver-staining technique

Abstract— A silver colloid technique to identify argyrophilic nucleolar organizer region associated protein (AgNOR) was applied to 43 cases of intrahepatic bile duct carcinoma (cholangiocarcinoma, CC), 2 with bile duct adenoma (BDA), 5 with focal duct epithelial hyperplasia (FEH) associated with hepatolithiasis, 15 with posthepatitic ductular proliferation (PHDP) associated with massive or submassive hepatic necrosis and 20 of normal liver. In the present study, only discrete, easily counted black dots within nuclei and silver-stained nucleolus were counted under a magnification of X 400 without oil-immersion objectives. The mean AgNOR count of CC was significantly higher than those of BDA, FEH, PHDP and normal controls (P>0.05, P>0.001, P>0.01, and P>0.001, respectively). Among CCs the mean AgNOR numbers of papillary adenocarcinoma (pap), moderately (tub2) and poorly differentiated (por) adenocarcinoma, and adeno-squamous carcinoma (as) were significantly higher than that of normal controls (P > 0.01, P > 0.001, P > 0.001 and P > 0.001, respectively), and those of tub2, por and as were also significantly higher than those of BDA, FEH and PHDP, whereas that of well differentiated tubular adenocarcinoma (tub 1) was not different from those of BDA, FEH, PHDP and normal controls, and that of pap was not different from those of BDA, FEH and PHDP. The mean numbers of AgNORs of BDA and FEH were not different from that of normal controls, whereas that of PHDP was significantly higher than that of normal controls (P > 0.01). Interestingly, the mean AgNOR counts of tubular adenocarcinoma were increased with histologic tumor grades. All cases with AgNOR counts of more than 2.24 had CC. Furthermore, not only quantitative but also striking qualitative abnormalities of AgNORs were seen in CCs. Large and/or irregularly shaped AgNORs without uniformity in size and density were characteristic for CCs, even in well differentiated adenocarcinoma, in contrast to the small, round, regular AgNORs seen in normal bile duct epithelium and rather large but regular AgNORs seen in benign bile duct lesions. These results indicate that the enumeration of AgNORs in various bile duct lesions is useful to identify moderate and high grade CCs and to evaluate tumor grade, but not useful to discriminate well differentiated adenocarcinoma from benign lesions. In case of normal or equivocal AgNOR counts, the presence of large and/or irregularly shaped AgNORs without uniformity in size and density would favor a diagnosis of carcinoma rather than benign counterparts.     

Prognostic value of silver-stained nucleolar organizer regions in colorectal carcinoma

AIM: Recently, silver-stained nucleolar organizing regions (AgNOR) analysis has been used as a criterion for tumor diagnosis and research. The purpose of this study was to investigate the prognostic value of AgNOR analysis in colorectal carcinomas. METHODS: The silver staining technique was applied to paraffin embedded tumor tissue sections from 114 patients with colorectal carcinoma. The number, morphology, size, and distribution of AgNOR were counted and analyzed. RESULTS: (1) The number of AgNOR in patients who died within 5 year of carcinoma diagnosis (x^-± s: 8.8 ± 2.3 per nucleus, n = 27) was significantly higher than that in those who survived beyond 5 year (6.3 ± 1.8, n = 30, p < 0.001). The number of large sized (> 2 μm) and small sized (< 1 μm) AgNOR was significantly higher in patients who died (x^-± s: 85.9 ± 20.7, 661.7 ± 250.5 in 100 nuclei) than in those who survived (71.7 ± 27.0, 398.3 ± 225.4, p = 0.04, 0.00 respectively). The concentrated type of distribution was significantly fewer in those who died (10.2%) than those who survived (31.4%, p = 0.00), whereas the mixed type of distribution was significantly greater in those who died (25.7%) than in those who survived (7.1%, p = 0.00). And (2) The number of AgNOR was also related to other factors that affected prognosis of colorectal carcinoma, such as age, histological type, depth of invasions, and metastasis to lymph nodes. CONCLUSION: The AgNOR analysis is a novel and useful parameter for assessing the prognosis of colorectal carcinoma.     

Changes of nucleolar organizer regions in granulopoietic precursors during the course of chronic myeloid leukemia

The aim of the present study was to analyze the nucleolar organizer region (AgNOR) pattern of granulopoietic precursors in chronic myeloid leukemia (CML) at diagnosis and during the course of the disease. Clusters of AgNORs and isolated dots were counted separately in 24 cases of CML at diagnosis, in 19 cases during the relapse of the chronic phase after treatment, and in 16 cases of blast crisis. For comparison, 20 cases of normal bone marrow were studied. Each cell type had its own characteristic AgNOR pattern, as has been described for normal bone marrow. There was no significant difference in the number of AgNORs between cells in the peripheral blood and bone marrow. Compared with normal granulopoiesis, myeloblasts in CML at diagnosis had lower numbers of clusters, which decreased further during relapse of chronic phase and in blast crisis. Promyelocytes and myelocytes showed significantly fewer dots. The number of AgNOR clusters correlates inversely with the duration of the cell cycle. Therefore, these findings are consistent with the progressive loss in proliferative activity of immature precursors described during the course of CML. As the number of dots indicates cellular maturation, their lower number in promyelocytes and myelocytes in CML favors the concept of a discordant maturation process described in this desease. The separate counting of clusters and dots provides a useful, simple, and cheap method of describing cytokinetic changes during the course of this myeloproliferative     

肺结核患者外周血T淋巴细胞亚群与核仁组成区嗜银蛋白的关系及临床应用的研究

目的　探讨肺结核患者T淋巴细胞亚群与核仁组成区嗜银蛋白(AgNORs)的关系,并动态观察临床应用的价值。方法 采用碱性磷酸酶抗碱性磷酸酶(APAAP)法及AgNORs测定方法,对98例肺结核患者,39例肺炎患者和41名正常人分别测定外周血T淋巴细胞亚群和AgNORs,并对71例初治肺结核患者进行了动态观察。结果 肺结核患者的CD₃⁺、CD₄⁺、CD₄⁺/CD₈⁺比值均明显降低,CD₈⁺升高;AgNORs的I.S%值明显降低,与健康人及肺炎患者相比有显著性差异(P<0.01)。随着肺结核患者病情的好转此两项指标逐渐上升。结论 肺结核患者T淋巴细胞亚群与AgNORs检测结果一致,可动态地反映结核病人的免疫状态,并与病情发展呈良好的相关性,可作为指导治疗的参考。     

Assessment of nucleolar organizer regions by automatic image analysis in breast cancer: correlation with DNA content,proliferation rate,receptor status and histopathological grading

Summary The value of automatic image analysis in the investigation of nucleolus regions (AgNOR) has been examined in tissue sections of 52 malignant and 30 benign breast lesions. Determination of the AgNOR number per cell alone revealed a considerable overlap between benign (range 1.2–3.8) and malignant specimens (range 1.5–16.2). They differed however, highly significantly (P<0.001) in their AgNOR sizes. In benign breast disorders the mean AgNOR area per tumour ranged from 0.22 m² to 1.07 m² (mean 0.39 m²), whereas in carcinomas AgNOR sites ranged from 0.05 m² to 0.22 m² (mean 0.09 m²). AgNOR counts showed a good correlation with histopathological grade (P<0.05), aneuploidy (P<0.01), proliferation rate as determined by Ki67 immunostaining (P<0.01), as well as oestrogen and progesterone receptor content (P<0.01). Image analysis proved to be advantageous over AgNOR counting alone as it facilitated the standardization of the AgNOR technique itself and thus, significantly improved its diagnostic specifity.Abbreviations AgNOR nucleolar organizer regions demonstrated by silver staining - Ki67 proliferation marker Dedicated to Professor Dr. D. Schmähl on the occasion of his 65th birthdaySupported in part by a grant from the Bundesminister für Forschung und Technologie and by the P. E. Kempes Foundation     

Argyrophilic nucleolar organizer regions in neoplastic and non-neoplastic hepatocytes bearing Mallory bodies

ABSTRACT— The proliferative activity of Mallory bodies (MB)-positive hepatocytes (neoplastic and non-neoplastic) was examined by counting the argyrophilic nucleolar organizer regions (AgNORs). Among 19 cases of hepatocellular carcinoma, the mean number of AgNORs was lower in the MB-positive carcinoma cells than in the negative ones in nine cases, higher in six, and there was no difference in four. In non-neoplastic cases (seven cases of advanced primary biliary cirrhosis and seven cases of alcoholic or nutritional liver injury), the mean number of AgNORs was lower in the MB-positive hepatocytes than that in the negative ones in eight cases, and approximately equal in number in six cases. These findings imply that MB formation does not directly represent the level of proliferative activity of hepatocytes, regardless of whether they are malignant or not.     

急性淋巴细胞白血病 T淋巴细胞核仁组成区相关蛋白的检测及临床意义

目的探讨T淋巴细胞核仁组成区嗜银蛋白(Ag-NORS)在急性淋巴细胞白血病( ALL)的诊断及病情监测方面的临床意义.方法采用KL型肿瘤免疫图像分析系统及细胞培养液,以银染色试剂及其方法对正常人及ALL患者不同临床时期外周血T淋巴细胞核进行银染色和分析.结果正常人T淋巴细胞Ag-NORS含量明显高于ALL患者(P<0.05),正常人与ALL患者中完全缓解(CR)者和未完全缓解(NR)者比较,差异均有显著性意义(分别为P<0.05和P<0.01).结论T淋巴细胞Ag-NORS检测在ALL诊断及病情监测方面具有重要意义.     

细胞核仁组成区嗜银蛋白检测对良恶性胸腔积液的鉴别诊断 …

目的 探讨核仁组成嗜银蛋白检测在良性胸腔积液间皮细胞和恶性胸腔积液癌细胞临别诊断中的价值。方法 对５０例恶怀、３０例良性胸腔积液鹗２的胸腔积液细胞涂片行ＡｇＮＯｒ染色，观察良性胸腔积液间皮细胞和恶性胸积液癌细胞核内的ＡｇＮＯＲ数目和形态。另观察６例临床疑诊恶性胸腔积液而常规脱落细胞检查阴性者胸积液细胞核内的ＡｇＮＯＲ数目和形态。结果 恶性组癌细胞平均每核ＡｇＮＯＲ数显著高于良性组间皮细胞；恶性组癌     

Identification of nucleolar organizer regions in non-neoplastic and neoplastic hepatocytes by the silver-staining technique

Abstract— The silver staining technique to demonstrate nucleolar organizer region (NOR)-associated proteins (AgNORs) was applied to a variety of liver tissues, including chronic persistent hepatitis (CPH), chronic active hepatitis (CAH), liver cirrhosis (LC), liver cell dysplasia (LCD), focal nodular hyperplasia (FNH), adenomatous hyperplasia (AH) and hepatocellular carcinoma (HCC). In the present study, only discrete, easily counted black dots within nuclei and silver-stained nucleolus were counted under a magnification of X 400 without oil-immersion objectives. The mean AgNOR counts of HCC and LCD were significantly higher than that of normal hepatocytes, and 77% of cases of LCD and 56% of HCC had mean AgNOR counts more than 2, whereas those in CPH, CAH, LC, FNH and AH were always less than 2 and were not different from that of normal hepatocytes. Among HCC, the mean number of AgNORs increased with the grade of the tumor. However, the AgNOR counts of grade I HCC were always less than 2 and overlapped with those of normal hepatocytes and other benign categories. All cases with mean AgNOR counts of more than 2 turned out to be HCC, except LCD which exhibited characteristic histologic appearances easily distinguished from HCC. These findings suggest that AgNORs could be quantitatively useful in evaluating the grade of HCC, even under routine microscopic examination without oil-immersion objectives, and mean AgNOR counts of more than 2 per nucleus are hallmarks of HCC.     

肺结核患者T淋巴细胞亚群,红细胞免疫功能与核仁组成区 …

目的 探讨结核病患者Ｔ淋巴细胞亚群，红细胞免疫功能及核仁组成相关嗜银蛋白（ＡｇＮＯＲｓ）的关系及其临床意义。方法 采用碱性磷酸酶抗碱性磷酸酶（ＡＰＡＡＰ）法，红细胞花环试验以及ＡｇＮＯＲｓ测定方法，对５９例肺结核患者和５０名正常对照组分别测定了外周血Ｔ淋巴细胞亚群，红细胞免疫Ｃ３ｂ受体花环率（Ｅ－Ｃ３ｂＲＲ）与红细胞循环免疫复合物花环率（Ｅ－ＩＣＲ），ＡｇＮＯＲｓ面积（ＧＡ），核面积（ＮＡ）及ＧＡ     

Argyrophilic nucleolar organizer regions in cells of thymoma and thymic carcinoma: correlation with DNA ploidy and clinicopathologic characteristics

STUDY OBJECTIVES: To evaluate the usefulness of argyrophilic nucleolar organizer region (AgNOR) counting and flow cytometric DNA analysis in the differential diagnosis of thymoma and thymic carcinoma, as well as in the differences among various stages and histologic subtypes of these tumors. DESIGN AND INTERVENTIONS: Paraffin-embedded blocks of 64 thymic epithelial tumors (20 noninvasive thymomas, 34 invasive thymomas, and 10 thymic carcinomas) were studied by AgNOR counting and flow cytometric DNA analysis. The thymomas were histologically classified as medullary, cortical, or mixed subtype. MEASUREMENTS AND RESULTS: Invasive thymomas had more AgNORs (-/+ SD) than noninvasive thymomas (7.93+/-2.90 vs 5.97+/-1.77; p < 0.01). The number of AgNORs of thymoma increased progressively with advances in stage (p < 0.01). Cortical thymomas had the highest number of AgNORs among the three subtypes (p < 0.05). Patients with thymoma who presented with myasthenia gravis also had a higher number of AgNORs (8.30+/-3.12 vs 6.50+/-2.03; p < 0.01). The AgNOR number did not correlate with the DNA ploidy of all specimens. CONCLUSIONS: AgNOR counting is useful in differentiating between invasive and noninvasive thymomas, and in predicting the stage of thymomas. A greater number of AgNORs was observed in patients with cortical thymoma and in those who presented with myasthenia gravis.     

T淋巴细胞Ag—NORs的检测在原发性肝癌诊断中的价值

目的 :研究T淋巴细胞核仁形成区嗜银蛋白 (Ag NORs)在原发性肝癌免疫学诊断中的意义。方法 :采用KL型肿瘤免疫计算机图像分析系统 ,测定 38例正常人和 2 5例原发性肝癌患者外周血T淋巴细胞Ag NORs的含量 ,以细胞核仁银染面积与细胞核面积比值 (I .S % )作为检测评估指标 ,以了解原发性肝癌患者Ag NORs的变化情况。同时 ,检测 2 5例原发性肝癌患者血AFP含量 ,以比较Ag NORs和AFP诊断原发性肝癌的灵敏性。结果 :2 5例原发性肝癌的平均I .S %值为 5 5 1%± 0 85 % ,明显低于正常人的 7 12 %± 0 78% ,二者差异显著 (P <0 0 5 ) ;Ag NORs的检测阳性率达 88% ,高于AFP的检测阳性率的 84% ,但差异无显著性 (P >0 0 5 )。 4例AFP正常的原发性肝癌患者中有 3例Ag NORs检测为异常。结论 :T淋巴细胞Ag NORs与原发性肝癌的诊断呈良好的相关性和较高的灵敏性 ,可望成为一种原发性肝癌免疫学诊断的指标     

Rapid detection of proliferating potential in human brain tumors by nucleolar organizer region staining on squash preparations

Summary Rapid detection of the proliferating potential of 37 human brain tumors was attempted using squash preparations stained by a silver colloid technique for argyrophilic protein associated with nucleolar organizer regions (AgNORs). Less than 1 h was required for staining. The mean number of AgNORs in cell nuclei of malignant or recurrent brain tumors (16 cases) including meningeal sarcoma, recurrent meningioma, recurrent craniopharyngioma, anaplastic astrocytoma, glioblastoma multiforme and metastatic brain tumor was 3.18, and the number for benign brain tumors (21 cases) including meningioma, neurinoma, pituitary adenoma, benign astrocytoma, ependymoma, and adenoma of lachrymal gland was 1.85. The former value was significantly greater than the latter value (P<0.001). These results indicate that quantitative analysis of AgNORs in brain neoplastic cells, using squash preparations, is useful to differentiate malignant from benign tumors within 1 h. Thus, this method provides rapid and useful information about the proliferative potential of human brain tumors even during operation.Abbreviation AgNOR argyrophilic protein associated with nucleolar organizer regions     

亚精胺对日本血吸虫成虫培养细胞核仁组织区相关嗜银蛋白的影响

目的以日本血吸虫培养细胞核仁组织区相关嗜银蛋白(Argyrophilicnucleolarorganizerregionassociatedproteins,AgNORs)为评价指标,探讨亚精胺(Spermidine,Spd)对培养细胞的促增殖作用。方法采用联合法将虫龄为24d的日本血吸虫成虫细胞接种于小盖玻片上,培养于常规培养基中。接种后第3天,随机分为实验组和对照组。将实验组细胞用无血清培养基配制的终浓度为75μmol/L的Spd处理48h,对照组细胞则用不含Spd的无血清培养基处理,时间相同;PBS清洗3次后换用常规培养基继续培养。于第7、14、21、28、35天分别对培养细胞进行AgNORs染色。使用HPIAS-2000图像分析系统,计算培养细胞核内AgNORs颗粒数及平均吸光值,并作统计分析。结果经Spd处理,各实验组培养细胞AgNORs着色明显深于对照组;但随着培养时间的延长,实验组与对照组细胞着色均逐渐变浅。各实验组细胞AgNORs颗粒数目与平均吸光值高于对照组,二者比较差异有统计学意义(u7=50.95、43.78;u14=55.23、43.54;u21=36.65、32.25;u28=31.6、32.95;u35=28.86、9.54;P<0.01)。各实验组AgNORs吸光值两两比较,除第7天与第14天之间差异无统计学意义外(q=0.008058,P>0.05),其余差异都有统计学意义(P<0.01)。结论Spd具有促进日本血吸虫培养细胞增殖的能力,每周用Spd处理培养细胞1次,能更大限度地促进血吸虫培养细胞的生长。     

Predictive value of argyrophilic nucleolar-organizer-region-associated proteins in bladder cancer, using cell-imprint preparations

The predictive value of quantifying argyrophilic nucleolar-organizer-region-associated proteins (Ag-NOR) in routinely processed paraffin-embedded tissue sections of bladder cancer remains controversial. The purpose of this study was to determine whether cell-imprint preparations facilitate a more accurate determination of the mean number of Ag-NOR per cell (Ag-NOR score). The utility of using the Ag-NOR scores of cell-imprint preparations as a prognostic indicator in patients with bladder cancer was explored. We evaluated 90 patients with newly diagnosed bladder cancer using cell-imprint preparations to determine Ag-NOR score. The score significantly correlated with known prognostic factors associated with this tumor, including histological grade (P<0.001), pathological stage (P<0.01), and papillary structure (P<0.001). Furthermore, the Ag-NOR score appeared to be an independent predictor of intravesical tumor recurrence of superficial bladder cancer (P<0.05). These results suggest that the Ag-NOR score determined in a cell imprint preparation may serve as a useful prognostic indicator in bladder cancer.Abbreviation Ag-NOR argyrophilic nucleolar-organizer-region-associated proteins     

Down-regulation of plasma membranous Annexin A1 protein expression in premalignant and malignant lesions of the oral cavity: correlation with epithelial differentiation

Purpose  To determine the potential involvement of ANXA1 in oral squamous-cell carcinoma (OSCC), we evaluated the ANXA1 protein expression in oral premalignant lesions (OPLs) and OSCCs and correlated the results with clinicopathologic variables. Methods  Matched normal and tumour specimens of 44 primary OSCCs and 28 OPLs were analyzed for ANXA1 subcellular localization and protein expression level by immunohistochemistry (IHC). Correlations between ANXA1-IHC staining scores of OSCCs and clinicopathologic features were evaluated by Fisher’s exact test. Results  Markedly down-regulation of ANXA1 protein expression was identified on the plasma membrane of epithelial cells in OSCCs (P < 0.001) and OPLs (P = 0.001) compared with normal counterparts. Moreover, loss of plasma membranous ANXA1 expression was significantly correlated with the poorly differentiated status of OSCC cells (P = 0.012). Conclusions  Our findings suggest that loss of ANXA1 is frequent and early event during oral carcinogenesis and that ANXA1 could contribute to maintaining epithelial differentiation in OSCC.     

CD34 expression in endothelial cells of small hepatocellular carcinoma: Its correlation with tumour progression and angiographic findings

The angiogenic process plays an important role in tumour growth and metastasis during hepatocarcinogenesis, but it is still uncertain when the process begins during tumour formation. Forty-two small hepatocellular carcinomas (HCC) that measured either less than or equal to 2 cm in diameter were studied by comparing the histologic findings with the angiographic findings, and with immunohistochemical expression of endothelial marker QB-end/10 (QB), a new monoclonal antibody raised against CD34, in the sinusoidal wall. Twenty (91%) of 22 moderately or poorly differentiated HCC revealed a positive reaction for QB, while only eight (40%) of 20 well differentiated HCC demonstrated a positive reaction (P < 0.01). In the tumours showing a ‘nodule in nodule’ appearance, the less differentiated areas were more reactive for QB. Twenty-three (82%) of 28 QB positive tumours were hypervascular, while only three of 14 (21%) QB negative tumours were hypervascular (P < 0.01) by angiography. All six of the poorly differentiated and 13 (81%) of the 16 moderately differentiated tumours were hypervascular, while only seven (35%) of 20 well differentiated HCC were hypervascular (P < 0.01). These results indicate that as the tumour becomes less differentiated, the QB positive areas become wider and angiography demonstrates hypervascularity. We therefore speculate that the HCC sinusoids acquire the characteristics of capillary and precapillary blood vessels during de-differentiation from well to moderate, and thus the tumour begins to reveal hypervascularity on angiography. The above process may be correlated with the stepwise progression of HCC.