Hepatocellular Nodular Lesions Research Articles

Deep Learning-Based Classification of Hepatocellular Nodular Lesions on Whole-Slide Histopathologic Images

Hepatocellular nodular lesions (HNLs) constitute a heterogeneous group of disorders. Differential diagnosis among these lesions, especially high-grade dysplastic nodules (HGDNs) and well-differentiated hepatocellular carcinoma (WD-HCC), can be challenging, letalone biopsy specimens. We aimed to develop a deep learning system to solve these puzzles, improving the histopathologic diagnosis of HNLs (WD-HCC, HGDN, low-grade DN, focal nodular hyperplasia, hepatocellular adenoma), and background tissues (nodular cirrhosis, normal liver tissue). The samples consisting of surgical and biopsy specimens were collected from 6 hospitals. Each specimen was reviewed by 2 to 3 subspecialists. Four deep neural networks (ResNet50, InceptionV3, Xception, and the Ensemble) were used. Their performances were evaluated by confusion matrix, receiver operating characteristic curve, classification map, and heat map. The predictive efficiency of the optimal model was further verified by comparing with that of 9 pathologists. We obtained 213,280 patches from 1115 whole-slide images of 738 patients. An optimal model was finally chosen based on F1 score and area under the curve value, named hepatocellular-nodular artificial intelligence model (HnAIM), with the overall 7-category area under the curve of 0.935 in the independent external validation cohort. For biopsy specimens, the agreement rate with subspecialists' majority opinion was higher for HnAIM than 9 pathologists on both patch level and whole-slide images level. We first developed a deep learning diagnostic model for HNLs, which performed well and contributed to enhancing the diagnosis rate of early HCC and risk stratification of patients with HNLs. Furthermore, HnAIM had significant advantages in patch-level recognition, with important diagnostic implications for fragmentary or scarce biopsy specimens.

Combined hepatocellular-cholangiocarcinoma and its mimickers: Diagnostic pitfalls in surgical pathology

BackgroundThe diagnosis of combined hepatocellular-cholangiocarcinoma (cHCC-CCA) requires histomorphological detection of both hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (iCCA). However, these primary liver cancers (PLCs) have a wide variety of microscopic appearances resulting in difficulties and uncertainties in cHCC-CCA's diagnosis. This study aims to perform a clinicopathologic analysis on the diagnosis of PLCs at a tertiary referral hospital in Thailand using traditional morphologic studies. MethodsA 5-year retrospective analysis of pathologically diagnosed PLCs was conducted. Pathological features and clinical characteristics of cHCC-CCA and other PLCs with the histopathologic resemblance to cHCC-CCA were studied. The pathological diagnosis was rendered based on histomorphological context rather than immunoreactivity. A literature review containing diagnostic pitfalls of cHCC-CCA was carried out. ResultsPLCs from a total of 295 patients were retrieved, and cHCC-CCA accounted for 1.4% (n = 4) of the malignancies. Histomorphological evaluation is the most reliable diagnostic modality for cHCC-CCA. Extremely uncommon variants of iCCA (i.e., mucinous iCCA and adenosquamous iCCA) and iCCA arising with hepatocellular nodular lesions (i.e., iCCA with nodular regenerative hyperplasia (NRH), and iCCA in cirrhosis) could have a histomorphologic resemblance to that of cHCC-CCA. ConclusionsAlthough there has been an exceedingly high incidence of iCCA in Thailand, such a commonness is not valid for cHCC-CCA in our series. Rare forms of iCCA could have a morphologic resemblance to that of cHCC-CCA. Regardless of the differentiation and immunophenotype, iCCA without a distinct HCC component should never be diagnosed as cHCC-CCA.

Diagnostic Role of Aldoketoreductase family 1B10 (AKR1B10) in Hepatocellular Carcinoma and Benign Hepatic Lesions

Abstract Background Hepatocellular carcinoma is the most common primary liver cancer. It is the second most common cause of cancer deaths. The majority of HCCs arise in chronically diseased livers with cirrhotic background. Several benign lesions might be found in a cirrhotic liver along with hepatocellular carcinoma (HCC), and may exhibit typical or atypical imaging features. Distinction between benign hepatocellular nodular lesions, such as hepatic adenoma, and focal nodular hyperplasia, from HCC is of crucial clinical importance in determining appropriate therapy and assessing the prognosis.AKR1B10 is a promising marker that is recently used in the HCC diagnosis. Aim of the Work is to study the immuno-histochemical expression of Aldoketoreductase family 1B10 (AKR1B10) in hepatocellular carcinoma and benign hepatic lesions, to assess its role in the diagnosis of hepatocellular carcinoma and in differentiating it from different benign hepatic lesions. Materials and Methods Hepatocellular carcinoma (HCC(T)):(109), corresponding non tumor tissue with and without cirrhosis (NT):(80), benign hepatic lesions (Focal Nodular Hyperplasia(FNH):(6) and Hepatic Adenoma(HCA)) (6) and Hepatic Cirrhosis (Cs) without HCC (16). Results AKR1B10 was over expressed in 82.6% (90/109) of studied HCC cases, negative expression in studied corresponding non-tumor hepatic tissue (NT) in 92.5% (74/80), complete negativity of AKR1B10 was observed in studied benign hepatic lesions 100% (0/12) and also negative expression in 81.3% (13/16) of included hepatic cirrhosis without HCC. Conclusion AKR1B10 is a promising marker in the diagnosis of HCC and distinguishing it from other benign hepatic lesions.

良性肝細胞性結節の病理診断：新WHO分類をふまえて

良性肝細胞性結節には様々な種類があり，従来から，その診断基準は詳細に設定されていた．しかし，肝細胞腺腫（HCA）に関して，2010年の新WHO分類により，肝細胞腺腫の分子病理学的性格を反映した免疫組織化学的診断法が導入されるようになった．そしてHCAは4つの亜型に分類され，また，それはFNHとの鑑別に有用であるとされた．本邦における症例でもその診断法は，HCAの診断，亜型分類，FNHとの鑑別にある程度有用であった．しかし，従来の疾患概念の定義や鑑別診断法と，新WHO分類とは，その診断根拠が異なり，その診断か必ずしも合致しない例も経験される．これらの状況をふまえ，良性肝細胞性結節の診断に関する，新しい考え方を提示する．

Fine needle aspiration biopsy of hepatocellular carcinoma and hepatocellular nodular lesions: role, controversies and approach to diagnosis

The role of fine needle aspiration (FNA) biopsy of the liver has evolved. Advances in imaging modalities have obviated the need for tissue confirmation in clinically classic hepatocellular carcinoma (HCC). The risks of needle tract seeding and haematogenous dissemination have been actively debated. Nowadays, cytopathologists are confronted by smaller and smaller nodules, detected due to increased surveillance of high-risk cirrhotic patients. Tissue characterization of small well-differentiated hepatocellular nodular lesions (size less than and equal to 2 cm) is extremely challenging and has therapeutic implications. Major issues in the cytodiagnosis of HCC include: (i) distinguishing benign hepatocellular nodular lesions, namely, large regenerative nodules, dysplastic nodules, focal nodular hyperplasia and hepatocellular adenoma from reactive hepatocytes; (ii) distinguishing well-differentiated HCC from benign hepatocellular nodular lesions; (iii) distinguishing poorly differentiated HCC from intrahepatic cholangiocarcinoma and metastatic carcinomas; (iv) determining the histogenesis of a malignant tumour; and (v) determining the site of origin of a malignant tumour. An overview of the biological evolution and histopathological aspects of dysplastic nodules, small HCCs and 'nodule-in-nodule' lesions is presented in tandem with clinically relevant nomenclature. An algorithmic approach to FNA diagnosis of HCC and hepatocellular nodular lesions is outlined. Optimal results depend on (i) a dedicated radiologist-cytopathologist team; (ii) an on-site cytology service, (iii) a combined cytohistological approach, (iv) immunohistochemistry, and (v) clinicopathological correlation. As we move towards personalized medicine, it is envisaged that hepatic FNA is likely to become a point of care in the management protocol as it takes on the additional role of procurement of tumour and peritumoural tissues for genomic and proteomic profiling to enable targeted molecular therapy.

Gadolinium Ethoxybenzyl Diethylenetriamine Pentaacetic Acid-Enhanced Magnetic Resonance Imaging Findings of Borderline Lesions at High Risk for Progression to Hypervascular Classic Hepatocellular Carcinoma

The objectives of the study were to assess the imaging features of hypovascular borderline lesions containing hypervascular foci on gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) and to evaluate the ability of Gd-EOB-DTPA-enhanced MRI to diagnose high-risk borderline lesions possibly consistent with early hepatocellular carcinoma (HCC). Institutional review board approval was obtained for this retrospective analysis of imaging findings, and informed consent was obtained from 217 consecutive patients undergoing Gd-EOB-DTPA-enhanced MRI and angiography-assisted computed tomography (CT) for examination of hepatocellular nodular lesions in cirrhotic livers. There were 73 nodules showing hypervascular foci in borderline lesions identified by angiography-assisted CT. Signal intensity patterns of the nodules were evaluated on hepatobiliary-phase Gd-EOB-DTPA-enhanced T1-weighted MRI obtained 20 minutes after intravenous injection of contrast media. Among 73 high-risk borderline lesions, 59 were hypointense (81%), and 14 were isointense (19%), compared with background liver parenchyma. There were 27 untreated lesions followed by CT and/or MRI. Almost half of these nodules transformed into hypervascular HCC, regardless of signal intensities seen on hepatobiliary-phase Gd-EOB-DTPA-enhanced MRI. Although many high-risk borderline HCC lesions are hypointense on hepatobiliary-phase Gd-EOB-DTPA-enhanced MRI, some high-risk borderline lesions are isointense and transform at the same rate into hypervascular HCC.

A pictorial review of benign hepatocellular nodular lesions: comprehensive radiological assessment incorporating the concept of anomalous portal tract syndrome

Although the pathological categorization system advocated by the International Working Party (IWP) on Terminology has been helpful in categorizing benign hepatocellular lesions, the diverse clinicopathological features of the lesions still cause confusion of diagnosis in clinical settings. Recently, an integrated disease concept termed "anomalous portal tract syndrome" (APTS) has been proposed as a congenital anomaly of the portal tract, being a single unifying etiological factor underlying the disorder. In this article, we discuss the radiological features of benign nodular hepatocellular lesions incorporated in the concept of APTS. We systematically reviewed the literature on benign hepatocellular lesions based on the concept of APTS, as well as standard IWP terminology. For this pictorial review, we selected six representative cases and assessed the radiological features of the cases based on the concept of APTS. The comprehensive assessment based on APTS enabled the systematic categorization of benign hepatocellular lesions, including nodular regenerative hyperplasia, large regenerative nodules, partial nodular transformation, focal nodular hyperplasia, and hepatocellular adenoma, and was helpful in understanding the overlapping features of these lesions. Although the disease concept of APTS is still evolving, it is nonetheless helpful in comprehensively understanding the clinicopathological and radiological features of various benign hepatocellular lesions.

Radiological and histopathological manifestations of hepatocellular nodular lesions concomitant with various congenital and acquired hepatic hemodynamic abnormalities

Congenital and acquired hepatic hemodynamic abnormalities are classified into four categories: hepatic arterial inflow disorder, portal vein inflow disorder, hepatic vein outflow disorder, and presence of a third inflow to the liver. Although their detailed etiology is not fully understood, these hepatic hemodynamic abnormalities may cause the formation of hepatocellular nodules. Recent advances in imaging modalities now enable visualization of these hepatocellular nodules concomitantly with the identification of various congenital and acquired hemodynamic abnormalities. Most of these nodular lesions are benign hyperplastic nodules, such as focal nodular hyperplasia, nodular regenerative hyperplasia, and other types of regenerative nodules. However, neoplastic nodules such as hepatic adenoma and hepatocellular carcinoma may also occur in conjunction with hepatic hemodynamic abnormalities. Distinguishing neoplastic nodules, especially malignant liver tumors, from hyperplastic nodules is important. Detection of intranodular Kupffer cells with superparamagnetic iron oxide enhanced magnetic resonance imaging is a key indicator that a nodule is regenerative rather than neoplastic.

A case of hypervascular hyperplastic nodules mimicking hepatocellular carcinoma in alcoholic liver cirrhosis

Benign hypervascular hyperplastic nodules (HHN) in liver cirrhosis are very rare. It is important to distinguish between regenerative nodules (hyperplastic nodules) and tumorous nodules (dysplastic or neoplastic nodules) in hepatocellular nodular lesions. The differential diagnosis between HHN and hepatocellular carcinoma on the basis of radiologic imaging is often difficult, and is clinically important when determining the therapeutic plan. Therefore, histological confirmation by needle biopsy sampling of the liver is necessary for a correct diagnosis of HHN. We report herein a case of benign HHN mimicking hepatocellular carcinoma in a 32-year-old male alcoholic liver cirrhosis patient without viral hepatitis infection.

Pathologic diagnosis of early hepatocellular carcinoma: A report of the international consensus group for hepatocellular neoplasia

Pathologic diagnosis of early hepatocellular carcinoma: A report of the international consensus group for hepatocellular neoplasia

Analysis of morphological variables and arterialization in the differential diagnosis of hepatic nodules in explanted cirrhotic livers

BackgroundMany terminologies have been given to dysplastic hepatocellular nodules, which are preneoplastic lesions. In 1995, the International Working Party meeting established the nomenclature and morphological criteria for hepatocellular nodular lesions. Nevertheless, an unequivocal differential diagnosis is sometimes difficult, particularly among large regenerative nodules, dysplastic nodules and hepatocellular carcinoma. Angiogenesis is observed during hepatocarcinogenesis and the presence of the isolated arteries may help to discriminate these nodules. The relevance of the International Working Party histological variables and presence of the isolated arteries were analyzed with regard to the diagnosis of large regenerative nodules, low and high grade dysplastic nodules and hepatocellular carcinoma, in order to evaluate which have a real contribution in such diagnoses.MethodsOne hundred and seven nodular hepatocellular lesions over 5 mm (or smaller nodules with a different color) from explanted cirrhotic livers were analyzed and classified following the criteria of the International Working Party. Classifications were as follows: large regenerative nodules, low grade dysplastic nodules, high grade dysplastic nodules and hepatocellular carcinoma. The presence of isolated arteries (not related to the portal tracts or fibrosis) was verified for the nodules.ResultsAmong the 107 nodular lesions studied, 17 were classified as large regenerative nodules, 38 as low grade dysplastic nodules, 28 as high grade dysplastic nodules and 24 as hepatocellular carcinoma. The most relevant International Working Party variables in the differential diagnosis of the nodules were cellularity, trabeculae thickness, cytoplasmic staining, nuclear atypia, pseudoacinar pattern, portal tracts, nucleocytoplasmic ratio and mitosis. The isolated arteries, identified by hematoxylin and eosin staining, were important discriminating between two groups: low grade lesions (large regenerative nodules/low grade dysplastic nodules) and high grade lesions (high grade dysplastic nodules/hepatocellular carcinoma) (P < 0.001).ConclusionThe International Working Party criteria allow for the classification of the majority of hepatocellular nodules. However, other features such as cytoplasmatic tintorial affinity and pseudoacinar pattern may contribute to these diagnoses. The finding of isolated arteries in a nodular lesion should be investigated carefully, since the nodule could be a dysplastic lesion or hepatocellular carcinoma.

Diagnostic Utility of Immunohistochemistry in Hepatocellular Carcinoma, its Variants and Their Mimics

Hepatocellular carcinoma (HCC) is known for its histomorphologic heterogeneity. Immunohistochemistry (IHC) can help in the comparative morphologic evaluation of HCC, its variants and their mimics. Some of these diagnostic challenges can be attributed to (i) the variety of neoplasms that can arise from the hepatic stem cell lineage; (ii) the spectrum of well-differentiated hepatocellular nodular lesions; (iii) the liver being a target for metastases with some of these histologic entities mimicking variants of HCC or actually arising in the liver; and (iv) the limitations of serum alpha-fetoprotein (AFP). The role of IHC is in the distinction of benign hepatocellular nodules from reactive hepatocytes; WD-HCC from benign hepatocellular nodules; poorly differentiated HCC from cholangiocarcinoma and metastases; and determination of histogenesis of malignant tumor; and of primary site of origin of malignant tumor. A panel of antibodies has more discriminant value. AFP expression usually indicates malignancy in a hepatocellular nodule and hepatocytic histogenesis of a malignancy. Polyclonal carcinoembryonic antigen (pCEA) and CD10 stain bile canaliculi in better-differentiated HCC. HepPar1 is generally accepted as a hepatocytic marker. However, not all HCC stain uniformly and not all HepPar1-positive tumors are of hepatocytic origin or arise in the liver. Mature hepatocytes and hepatocellular nodules stain with CAM 5.2, CK 8, and 18 but not with CK 7, 19, 20, or AE1/AE3. Biliary epithelium expresses CK 7 and 19. CD 34 highlights sinusoidal capillarization. AFP, pCEA/CD10, and CD34 are useful for ascertainment of malignancy in hepatocellular nodules; HepPar1 and cytokeratins to be included if histogenesis is the issue. IHC results should be interpreted in the larger context of the case.

Molecular changes from dysplastic nodule to hepatocellular carcinoma through gene expression profiling

Progression of hepatocellular carcinoma (HCC) is a stepwise process that proceeds from pre-neoplastic lesions--including low-grade dysplastic nodules (LGDNs) and high-grade dysplastic nodules (HGDNs)--to advanced HCC. The molecular changes associated with this progression are unclear, however, and the morphological cues thought to distinguish pre-neoplastic lesions from well-differentiated HCC are not universally accepted. To understand the multistep process of hepato-carcinogenesis at the molecular level, we used oligo-nucleotide microarrays to investigate the transcription profiles of 50 hepatocellular nodular lesions ranging from LGDNs to primary HCC (Edmondson grades 1-3). We demonstrated that gene expression profiles can discriminate not only between dysplastic nodules and overt carcinoma but also between different histological grades of HCC via unsupervised hierarchical clustering with 10,376 genes. We identified 3,084 grade-associated genes, correlated with tumor progression, using one-way ANOVA and a one-versus-all unpooled t test. Functional assignment of these genes revealed discrete expression clusters representing grade-dependent biological properties of HCC. Using both diagonal linear discriminant analysis and support vector machines, we identified 240 genes that could accurately classify tumors according to histological grade, especially when attempting to discriminate LGDNs, HGDNs, and grade 1 HCC. In conclusion, a clear molecular demarcation between dysplastic nodules and overt HCC exists. The progression from grade 1 through grade 3 HCC is associated with changes in gene expression consistent with plausible functional consequences.

Fine needle aspiration biopsy of the liver: Algorithmic approach and current issues in the diagnosis of hepatocellular carcinoma.

The role of fine needle aspiration biopsy (FNAB) in the evaluation of focal liver lesions has evolved. Guided FNAB is still useful to procure a tissue diagnosis if clinical, biochemical and radiologic findings are inconclusive. Major diagnostic issues include: (i) Distinction of benign hepatocellular nodular lesions from reactive hepatocytes, (ii) Distinction of well-differentiated hepatocellular carcinoma (WD-HCC) from benign hepatocellular nodular lesions, (iii) Distinction of poorly differentiated HCC from cholangiocarcinoma and metastatic carcinomas, (iv) Determination of histogenesis of malignant tumor, and (v) Determination of primary site of origin of malignant tumor. This review gives a general overview of hepatic FNAB; outlines an algorithmic approach to cytodiagnosis with emphasis on HCC, its variants and their mimics; and addresses current diagnostic issues. Close radiologic surveillance of high-risk cirrhotic patients has resulted in the increasing detection of smaller lesions with many subjected to biopsy for tissue characterization. The need for tissue confirmation in clinically obvious HCC is questioned due to risk of malignant seeding. When a biopsy is indicated, core needle biopsy is favored over FNAB. The inherent difficulty of distinguishing small/early HCC from benign hepatocellular nodular lesions has resulted in indeterminate reports. Changing concepts in the understanding of the biological behavior and morphologic evolution of HCC and its precursors; and the current lack of agreement on the morphologic criteria for distinguishing high-grade dysplastic lesions (with small cell change) from WD-HCC, have profound impact on nomenclature, cytohistologic interpretation and management. Optimization of hepatic FNAB to enhance the yield and accuracy of diagnoses requires close clinicopathologic correlation; combined cytohistologic approach; judicious use of ancillary tests; and skilled healthcare teams.

Mutations of β‐catenin and AXIN I genes are a late event in human hepatocellular carcinogenesis

Hepatocellular carcinoma (HCC) is a well-known cancer involving the Wnt pathway in its carcinogenesis. However, it is not clear whether these genetic changes are early genetic events in hepatocarcinogenesis or not. In this study, we performed mutational analysis of the beta-catenin and AXIN I genes, and immunohistochemistry for beta-catenin in a series of 114 hepatocellular nodular lesions, including premalignant lesions such as low-grade dysplastic nodules (LGDNs) and high-grade dysplastic nodules (HGDNs). In the present study, mutations of the beta-catenin and AXIN I genes were detected in 16% (13 out of 81) and 6.2% (five of 81) of the HCCs, respectively. However, no mutations were found in 14 LGDNs and 19 HGDNs. Moreover, abnormal nuclear beta-catenin immunostaining was observed in 30 of 81 HCCs, but not in dysplastic nodules. Taken together, our data suggest that beta-catenin stabilization because of either beta-catenin or AXIN I mutation might be a late event for malignant progression rather than an early genetic event involving the initiation of HCC development.

Benign versus malignant hepatic nodules: MR imaging findings with pathologic correlation.

According to the currently used nomenclature, there are only two types of hepatocellular nodular lesions: regenerative lesions and dysplastic or neoplastic lesions. Regenerative nodules include monoacinar regenerative nodules, multiacinar regenerative nodules, cirrhotic nodules, segmental or lobar hyperplasia, and focal nodular hyperplasia. Dysplastic or neoplastic nodules include hepatocellular adenoma, dysplastic foci, dysplastic nodules, and hepatocellular carcinoma (HCC). Many of these types of hepatic nodules play a role in the de novo and stepwise carcinogenesis of HCC, which comprises the following steps: regenerative nodule, low-grade dysplastic nodule, high-grade dysplastic nodule, small HCC, and large HCC. State-of-the-art magnetic resonance (MR) imaging facilitates detection and characterization in most cases of hepatic nodules. State-of-the-art MR imaging includes single-shot fast spin-echo imaging, in-phase and opposed-phase T1-weighted gradient-echo imaging, T2-weighted fast spin-echo imaging with fat saturation, and two-dimensional or three-dimensional dynamic multiphase contrast material-enhanced imaging.

Conspicuity of hepatocellular nodular lesions in cirrhotic livers at ferumoxides-enhanced MR imaging: importance of Kupffer cell number.

To correlate the conspicuity of hepatocellular carcinomas and dysplastic nodules on ferumoxides-enhanced magnetic resonance (MR) images with the number of Kupffer cells in the hepatic lesions, as compared with that in background liver in histopathologic findings. Sixty-nine histopathologically proved moderately or poorly differentiated hepatocellular carcinomas, 10 well-differentiated hepatocellular carcinomas, and 19 dysplastic nodules were retrospectively studied in 68 patients with cirrhosis who underwent ferumoxides-enhanced MR imaging. The contrast-to-noise ratio between the nodules and surrounding parenchyma was calculated at T2-weighted fast spin-echo imaging, and the difference in the number of Kupffer cells between the nodules and surrounding hepatic tissue was calculated histopathologically. The results of MR imaging and histopathologic examination were correlated. All 69 moderately or poorly differentiated hepatocellular carcinomas had high contrast-to-noise ratios at MR imaging and large differences in the number of Kupffer cells. Six of the 10 well-differentiated hepatocellular carcinomas had contrast-to-noise ratios of zero or nearly zero, and five of these had little difference in the number of Kupffer cells. All 19 dysplastic nodules had contrast-to-noise ratios of zero or nearly zero, and there were virtually no differences in the number of Kupffer cells. Hepatocellular nodule conspicuity at ferumoxides-enhanced MR imaging depends on differences in the number of Kupffer cells within a nodule and the surrounding cirrhotic liver; moderately or poorly differentiated hepatocellular carcinomas can be distinguished from well-differentiated hepatocellular carcinomas and dysplastic nodules.

Histopathological and morphometric analysis of atypical adenomatous hyperplasia of human cirrhotic livers.

Atypical adenomatous hyperplasia (AAH) is a hyperplastic parenchymal nodular change in the cirrhotic liver, in which overt hepatocellular carcinoma (HCC) occasionally arises. AAH is defined as a sizable hepatocellular nodule with a variable degree of hepatocellular atypia not regarded as HCC, and is different from ordinary adenomatous hyperplasia in which hepatocellular atypia is absent. In the present study, we attempted to evaluate carcinogenetic processes and to find histological variables which indicate malignant transformation in AAH, using 49 surgically resected or autopsied nodules. AAH frequently showed morphological heterogeneity. Atypical lesions within AAHs were divisible into the following three categories from overall histopathological appearances: malignant (A), equivocal (B), or non-malignant (C) lesions. Analysis of combination of these three lesions, which were frequently intermixed in a given AAH, suggested that B lesions appear subsequent to C lesions, and A lesions finally appear in AAH nodules. Among the 14 histological variables, enlargement, hyperchromasia and irregular contour of nuclei were found to correlate well with A lesions. Increased nuclear density, iron resistance, reduction of reticulin fibres, clear cell change, sinusoidal dilatation and presence of abnormal arteries were suggestive of A or B lesions. Nuclear deviation toward the sinusoids, acinar and compact arrangements, fatty change and Mallory's hyaline alone were not useful indicators of A or B lesions. These results indicate that AAH is a preneoplastic or borderline lesion in which overt HCC is likely to evolve through several steps. Although a needle liver biopsy is a useful tool for diagnosis of benign, equivocal and malignant hepatocellular nodular lesions, the needle biopsy specimen should be carefully evaluated by considering the morphological heterogeneity of the AAH and a variable combination of 14 histological variables.