Banff classification and progress on liver allograft pathology / 器官移植

The pathology of liver allograft biopsy is not only essential for the evaluation of liver donor, but also for the diagnosis and differential diagnosis of posttransplantation complications. With the development of liver transplantation in clinical practice, relevant studies of the pathological diagnosis of liver allograft complications have been deepened. Banff classification on liver allograft pathology have been gradually established within the international community. In China, pathological studies related to liver allograft pathology have been steadily carried out, and the pathological diagnostic basis of liver allograft pathology suitable for the clinical practice of liver transplantation in China has been gradually formed. This article reviews the history of Banff liver allograft pathology and major pathological lesions of liver allograft complications, aiming to provide reference for implementing pathological diagnosis of liver allograft pathology in China, assisting clinical diagnosis and targeted treatment of complications after liver transplantation, and further improving the survival of liver allograft and recipients.

Advances in the diagnostic pathological features of precancerous lesions of hepatocellular carcinoma / 中华肝脏病杂志

The occurrence of hepatocellular carcinoma (HCC) is a multistep development process through precancerous lesions. A precancerous lesion of HCC is classified into hepatocyte dysplasia at the cytological level and dysplastic nodules at the histological level, and the corresponding lesion subtypes have different risks of canceration. Pathology is the "gold standard" for the diagnosis of early stage HCC and its precancerous lesions. However, it also faces many difficulties and challenges, such as the accumulation of experience in the pathological diagnosis, the understanding and grasp of key points of histopathological diagnosis and differential diagnosis, the combination application of immune and molecular diagnostic markers, and many others. This article briefly discusses the key points of pathological features and differential diagnosis of precancerous lesions of HCC.

Make great efforts to improve the levels of refined diagnosis and individualized treatment for microvascular invasion of liver cancer in China / 中华肝胆外科杂志

Recurrence after surgical resection is a bottleneck that severely restricts the long-term efficacy of patients with liver cancer, mainly referring to hepatocellular carcinoma and intrahepatic cholangiocarcinoma. As the most representative pathobiological feature of liver cancer, microvascular invasion (MVI) should be taken as an important focus to break through the bottleneck of postoperative recurrence. The new pathological diagnostic criteria and classification scheme of MVI has been written into the current " Guidelines for Diagnosis and Treatment of Primary Liver Cancer" of China. It is not only an important pathological indication for the clinical implementation of anti-recurrence therapy after operation, but also an key pathological basis for the clinical formulation of postoperative anti-recurrent therapy. Therefore, we intend to investigate how to make better use of it. From now on, multidisciplinary research should be strengthened in order to make real progress in the occurrence mechanism, migration pathway, distribution pattern, accurate identification and effective treatment of MVI in order to further improve the long-term efficacy of patients with liver cancer.

Make great efforts to improve the levels of refined diagnosis and individualized treatment for micro-vascular invasion of liver cancer in China / 中华肝胆外科杂志

Recurrence after surgical resection is a bottleneck that severely restricts the long-term efficacy of patients with liver cancer, mainly referring to hepatocellular carcinoma and intrahepatic cholangio-carcinoma. As the most representative pathobiological feature of liver cancer, microvascular invasion ( MVI) should be taken as an important focus to break through the bottleneck of postoperative recurrence. The new pathological diagnostic criteria and classification scheme of MVI has been written into the current"Guidelines for Diagnosis and Treatment of Primary Liver Cancer" of China. It is not only an important pathological indi-cation for the clinical implementation of anti-recurrence therapy after operation, but also an key pathological basis for the clinical formulation of postoperative anti-recurrent therapy. Therefore, we intend to investigate how to make better use of it. From now on, multidisciplinary research should be strengthened in order to make real progress in the occurrence mechanism, migration pathway, distribution pattern, accurate identifi-cation and effective treatment of MVI in order to further improve the long-term efficacy of patients with liver cancer.

Copy number aberrations of genes related to extrohepatic metastasis-free survival after operation for hepatocellular carcinoma / 中华肝脏病杂志

Objective@#To investigate the molecular markers of copy number aberrations (CNAs) of genes related to extrohepatic metastasis-free survival after the operation for hepatocellular carcinoma (HCC).@*Methods@#The CNA status of 20 candidate genes in 66 HCC samples was detected by microarray comparative genomic hybridization. The associations between gene CNAs and extrohepatic metastasis-free survival were evaluated using the Cox regression model, Log-rank test, and Kaplan-Meier survival analysis.@*Results@#Multivariate Cox analysis revealed that the independent risk factors for metastasis-free survival were MDM4 gain (hazard ratio [HR] = 2.74, 95% confidence interval [CI] = 1.18-6.37, P < 0.05), APC loss (HR = 8.43, 95% CI = 2.48-28.66, P < 0.01), and BCL2L1 gain (HR = 3.45, 95% CI = 1.13-10.52, P < 0.05) and the independent protective factor was FBXW7 loss (HR = 0.32, 95% CI = 0.12-0.89, P < 0.05). By stepwise Cox regression analysis, three CNAs related to metastasis-free survival were screened out: MDM4 gain (HR = 2.71, 95% CI = 1.11-6.64, P < 0.05), APC loss (HR = 7.19, 95% CI = 1.88-27.60, P < 0.005), and FBXW7 loss (HR = 0.16, 95% CI = 0.05-0.46, P < 0.01). There were significant differences in metastasis-free survival rate between the HCC patients with FBXW7 loss and without MDM4 gain or APC loss, those with MDM4 gain and/or APC loss and without FBXW7 loss, and those with other CNA combinations (log-rank test, P < 0.01).@*Conclusion@#MDM4 gain, APC loss, and FBXW7 loss are the independent prognostic factors for extrohepatic metastasis-free survival after the operation for HCC and can be used to predict the risk of extrohepatic metastasis after the operation for HCC.

Association of chromosome 17q copy number variation with overall survival of patients with hepatocellular carcinoma and screening of potential target genes / 中华医学遗传学杂志

OBJECTIVE To assess the association of copy number variations (CNVs) in chromosome 17q with the overall survival(OS) of patients with hepatocellular carcinoma(HCC), and to screen for target genes contained in the OS-related CNVs. METHODS A total of 174 HCC cases were enrolled. For 66 patients, the follow-up data was available. High-resolution Agilent Hu-244A array comparative genomic hybridization (aCGH) and Affymetrix U133 Plus 2.0 expression arrays were used to detect CNVs and gene expression of genes from the 17q region, respectively. The association of CNVs and OS was assessed with Log-rank test, Kaplan-Meier survival analysis, and Cox proportional hazards models. The gene expression in HCCs with 17q gain, HCCs without, and non-tumor liver tissues were compared with a Mann-Whitney U test. RESULTS Univariate association analysis showed that copy number gain in 17q25.1-25.3 was significantly associated with reduced OS (Log-rank test, P = 0.00002), and HCC cases with 17q25.1-25.3 gain had a 4.76-fold (95%CI: 2.31-9.81) increased hazard ratio (HR) for death from HCC, as compared to those without the gain. Multivariate Cox proportional hazards regression model revealed 17q25.1-25.3 gain to be an independent prognostic marker for poor OS (HR = 3.17, 95%CI: 1.39-7.26, P = 0.006). The expression levels of 18 genes in 17q25.1-25.3 including SLC9A3R1, GRB2, and TK1 were significantly increased in HCCs with gain than in those without (all P < 0.01) and non-tumor liver tissues (all P < 0.01). CONCLUSION The association of 17q25.1-25.3 gain with reduced OS has indicated that it is a prognostic marker for poor patient survival in HCC, for which SLC9A3R1, GRB2, and TK1 are candidate genes.

Copy number alterations and target genes in chromosome 6 short arm were related to postoperative recurrence of patients with hepatocellular carcinoma / 中华肝胆外科杂志

Objective To investigate the relationship between chromosome 6p copy number alterations (CNAs) and postoperative intrahepatic recurrence of hepatocellular carcinoma (HCC);and to screen for the target genes in CNA(s).Methods Array comparative genomic hybridization (CGH) and expression arrays were used to detect CNAs and differences in gene expression, respectively.The associations between CNAs in 6p and HCC recurrence were analyzed using the log-rank test, the Kaplan-Meier curves and the Cox proportional hazards models on 66 patients who had been follow-up for 2.6 ～ 73.3 months.The differentially expression of genes in the potentially recurrence-related CNAs were further evaluated by the MannWhitney U test on 117 HCCs, which included 109 cases with paired array CGH and expression data.Results 6p CNAs were detected in 46 (69.7％) of the 66 HCCs.Of the 8 CNAs with the most frequent recurrence of over 20％ , a gain at 6p21.1 was independently associated with a 2.3-fold (95％ CI =1.1 ～ 5.1, P ＜ 0.05) increased risk for intrahepatic recurrence and with a more pronounced 3.3-fold (95％ CI =1.4 ～ 8.2, P ＜0.05) risk for early recurrence (≤ 1 year).A panel of 9 genes, including BYSL and RPL7L1 within the documented 6p21.1, were observed to be upregulated in HCCs with 6p21.1 gain when compared with HCCs without (all P ＜ 0.05).A high BYSL expression significantly correlated with a larger tumor size (＞ 6 cm), vascular invasion and advanced tumor stage (all P ＜ 0.05), and high RPL7L1 expression significantly correlated with vascular invasion and advanced tumor stage (all P ＜ 0.05).Conclusion A gain at 6p21.1 was an independently prognostic marker for intrahepatic recurrence of postoperative HCC, particular for early recurrence, and BYSL and RPL7L1 might be the target genes in the recurrence-related 6p21.1 gain.

Association of chromosome 8p copy number alteration with postoperative survival of patients with hepatocellular carcinoma and screening of potential target genes / 中华肝胆外科杂志

Objective To investigate the association of chromosome 8p copy number alteration (CNA) with postoperative survival of patients with hepatocellular carcinoma (HCC),and to screen for possible target genes in the survival-related CNA (s) in 8p.Methods 187 HCC patients were enrolled into the study,which included 66 patients whose follow-up data were available and the follow-up was 2.6 ～ 73.3 months.High-resolution Agilent 244K comparative genomic hybridization (CGH) and Affymetrix U133 Plus2.0 expression arrays were used to screen for CNAs and gene expression differences in 8p.The associations between CNAs in 8p and survival were analyzed using the log-rank test,Kaplan-Meier survival analysis and Cox proportional hazards models.The gene expression levels between the groups were compared by the Mann-Whitney U test.Results Copy number loss on 8p12 (31/66,47％) was significantly associated with reduced survival rate,and HCC patients with 8p12 loss had a 4.1-fold (95％ CI =1.8 ～ 9.4,P ＜ 0.05) increased hazard ratio (HR) for death from HCC,as compared to those without the loss.The mRNA expression levels of the 3 genes in 8p12,including TMEM66,DCTN6,and MAK16,were significantly decreased in HCCs with gene loss than in HCCs without the loss (all P ＜ 0.05),and in non-tumorous liver tissues (all P ＜ 0.05).Conclusion Loss of 8p12 is an independent prognostic marker of unfavorable survival for patients with HCC,and underexpression of genes TMEM66,DCTN6,and MAK16,owing to 8p12 loss,contributed to unfavorable prognosis.

Combined effects of p53 and MDM2 polymorphisms on susceptibility and surgical prognosis in hepatitis B virus-related hepatocellular carcinoma

The p53 signaling pathway works as a potent barrier to tumor progression. Two single nucleotide polymorphisms (SNPs) in the gene loci of p53 pathway, p53 codon 72 Arg72Pro and MDM2 SNP309 (T > G), have been shown to cause perturbation of p53 function, but the effect of the two SNPs on the risk of hepatocellular carcinoma (HCC) remains inconsistent. This study investigated the influence of combined p53 Arg72Pro and MDM2 SNP309 on the risk of developing HCC in patients with chronic hepatitis B virus infection, and evaluated the significance of the two combined SNPs on patient prognosis. In total, 350 HCC patients, 230 non-HCC patients, and 96 healthy controls were genotyped for the p53 Arg72Pro and MDM2 SNP309. The combined p53 Pro/Pro and MDM2 G/G genotype was significantly associated with HCC risk (P = 0.047). Multivariate analysis indicated that combined p53 Pro/Pro and MDM2 G/G genotype was an independent factor affecting recurrence and survival (P < 0.05). Patients with combined p53 Pro/Pro and MDM2 G/G genotypes had a poorer prognosis than other genotypes, P < 0.01 for both disease-free survival (DFS) and overall survival (OS). DFS and OS rates also differed significantly between Barcelona Clinic Liver Cancer (BCLC) stage A patients with combined p53 Pro/Pro and MDM2 G/G and other genotypes (P < 0.05). Thus, the combined p53 Pro/Pro and MDM2 G/G genotype is associated with increased risk of developing HCC and is an independent adverse prognostic indicator in early stage HCC.

Some thoughts on the development of surgical pathology for hepatobiliary tumors / 中华肝胆外科杂志

Surgical-oncological pathology is one of the most important supporting discipline for hepatobiliary surgery.With rapid development in modern hepatobiliary surgery,the traditional model of morphological diagnosis can no longer meet the needs of oncological surgery.To improve the level of pathological diagnosis, better understand clinical concerns,update the mode of pathological diagnosis,introduce new technologies,expand the connotations of pathological diagnosis,provide a new indexing system for clinical treatment programs,and focus on interdisciplinary collaborative research are the most important issues in the development of surgical pathology for hepatobiliary tumors.

Progress and prospect on the clinicopathological study of small hepatocellular carcinoma / 中华肝胆外科杂志

Early detection and treatment of hepatocellular carcinoma (HCC) is one of the most direct and effective ways to improve the long-term outcome of patients with HCC. The comparative study on clinicopathological characteristics of surgically resected micro HCC (≤1 cm), small HCC(≤3 cm), middle HCC (3-5 cm), and large HCC (>5 cm) showed that ≤3 cm small HCC had the features of DNA diploidy, well differentiation, relatively slow growth, encapsulation, rare long-distance metastatic foci and thrombosis, easy to be radically resected and long-term postoperative survival. It is suggested that HCC growing to about ≤3 cm is an important growth phase when the change of pathobiological characteristics of HCC from a relatively benign to a more malignant behavior would occur. This is also the important time to carry out radical treatment to get better results. Therefore,further research on the early diagnosis and treatment as well as the pathobiological features of small HCC ≤3 cm should be undertaken as one of the key clinical and pathological studies in the future.

Immunopathological features of hepatic angiomyolipoma: An analysis of 36 cases / 第二军医大学学报

To study the immunopathological characteristics and differential diagnosis of hepatic angiomyolipoma(AML).Methods:Thirty-six surgically resected hepatic AML were investigated clinicopathologically and immunohistochemically with 10 antibodies.Results:Hepatic AML occurred in 21 females and 15 males,with the mean age of 41.6 years(26-60 years old).The patients with AML often had no special symptoms even had large space-occupying lesions in the liver.The diameter of AML was 2.5 cm to 14 cm(mean 6.8 cm).Histologically,AML was composed of varying heterogeneous mixture of 3 tissue components:blood vessels,smooth muscle and adipose cells.Extramedullary hemopoiesis sometimes existed.According to tissue components,AML was subcategorized into mixed type(19.4%,n=7),lipomatous type(11.1%,n=4),myomatous type(66.7%,n=24),and angiomatous type(2.8%,n=1).The epithelioid smooth muscle cells were sensitive to HMB-45(100%),SMA(100%),and CD117(66.7%) staining.Conclusion:Hepatic AML often contains smooth muscle elements,which have varied morphological features and should be carefully differentiated from hepatocellular carcinoma,mesenchymal hamartoma，and tumors with rich fat or blood vessels.Immunohistochemical staining with HMB-45 and SMA are the best available markers for the diagnosis of hepatic AML.

Immunohistochemical spectrum in the detection and differentiation of intrahepatic neoplasms / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To scrutinize the immunohistochemical spectrum to differentiate hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma (ICC) and metastatic adenocarcinoma (MAC) in the liver.</p><p><b>METHODS</b>Seven antibodies including AFP, Hep Par 1, CK18, CK19, CA19-9, CD34 and pCEA were immunohistochemically stained in resected specimens of 300 HCC, 35 ICC and 30 MAC. The specificity and sensitivity of the antibodies were evaluated by comprehensive capability score (CCS), with only those with CCS > or = 8 considered as having highly diagnostic value.</p><p><b>RESULTS</b>Antibodies CCS > or = 8 were observed as Hep Par 1 and CD34 in HCC, and CK19 in ICC, but none in MAC. For HCC, CCS of Hep Par 1 was higher than that of AFP (9 vs. 7) with 83.7% in sensitivity and 96.7% in specificity.</p><p><b>CONCLUSION</b>For HCC, Hep Par 1 and CD34 can be used as the first line antibodies, AFP and pCEA as the second line ones. CK19 is the first line antibody for ICC, and CA19-9 as the second. Hep Par 1, CD34 and CK19 are definitely helpful for the routine immunohistochemical stain to differentiate HCC from ICC and MAC.</p>

Surgical treatment of primary hepatic lymphoma: report of five cases / 中国普通外科杂志

Objective To investigate the clinicopathological features of primary hepatic lymphoma (PHL).Methods Data of 5 PHL cases treated at our hospital between 1998 and 2002 were retrospectively analyzed. Results The patients were all male with a mean age of 43 6 years. Abdominal pain and fever were the most frequent presenting symptoms. Four (80%) were positive for the hepatitis B virus antigen. Tumors were all removed surgically in the 5 patients. Pathologic examination revealed B cell lymphoma, T cell lymphoma and Hodgkin′s disease in 2, 2 and 1 patients, respectively. Moreover, coexistent tumors were incidentally discovered by histology in 2 cases. ConclusionThere seems to be a close relationship between hepatitis B virus infection and the development of PHL; PHL should always be considered in the differential diagnosis for patients presenting with mass lesions in the liver; Surgical resection whenever feasible should be attempted as a part of combination modalities.

Biological characters of hepatitis B virus transgenic mice strain C57-TgN(HBV adr2. 0)SMMU / 第二军医大学学报

Objective:To evaluate the biological characters of C57-TgN(HBV adr2.0)SMMU transgenic mice. Methods: Integration,expression,replication and histology change of hepatitis B virus gene in F6 transgenic mice were estimated by ge-nomic DNA PCR,Western blotting,ELISA,immunohistochemistry,serum DNA PCR,transmission electron microscopy and H-E staining. Results: Hepatitis B virus gene was integrated into F6 C57-TgN(HBV adr2. 0)SMMU transgenic mice and expressed HBsAg,HBcAg and X protein in liver tissue. HBsAg and HBeAg were expressed in serum of 19. 54% and 3. 39% F6 transgenic mice. Hepatitis B virus were replicated in serum and liver tissue of transgenic mice. Long-term integration,expression and replication of hepatitis B virus gene induced pathological lesion of transgenic mice liver and lung. Conclusion: C57-TgNCHBV adr2. 0)SMMU transgenic mice line has the biological characters including integration of hepatitis B virus gene into genomic DNA,expression and replication of hepatitis B virus gene in serum and liver, and histological change in liver and lung. It is a valuable animal system to study pathogenesis, treatment and prevention of hepatitis B virus.

Clinicopathological study on primary malignant vascular tumors of liver / 第二军医大学学报

Objective:To study the clinicopathological and immunohistochemical features of hepatic malignant vascular tumors. Methods:H E and immunohistochemistry were used to study 6 angiosarcoma and 5 epithelioid hemangioendothelioma (EHE) of the liver. Patients were aged 0.5 68 years, 7 were females and 4 were males. Results:Patients presented with nonspecific symptoms. Macroscopically,the tumors were usually multiple (3 13.7 cm in diameter) and were frequently hemorrhagic and multinodular. Histologically,there was a wide morphologic spectrum among cases. Six angiosarcoma included areas similar to cavernous and capillary hemangioma,spindle cell and epithelioid sarcoma. In the hepatic EHE,the tumor were comprised of dendritic and epithelioid cells that often contained vacuoles representing intracellular lumina. The stroma was fibrous with myxohyaline areas. Immunohistochemically,all tumors were positive for at least one endothelial marker(factor Ⅷ related antigen and /or CD 34 ). Conclusion:Hepatic malignant vascular tumor is very rare, the angiosarcoma of the liver is highly malignant with short survival. The prognosis of EHE is usually much better than that of angiosarcoma, surgical resection can obtain prolonged survival. The diagnosis can be made based on histological and immunohistochemical features of the tumor.

A study on imagination features of tumor thrombi in the portal vein of primary liver cancer / 中国普通外科杂志

Objective To study on imagination features of tumor thrombi in the portal vein of primary liver cancer (PLC). Methods We established a new type system of tumor thrombi, with Ⅰ 0 to Ⅳ, a total of 5 types and 8 sub-types in the portal vein of PLC based on normal intrahepatic portal vein anatomy and growing features of the tumor thrombi. The relationship between the types and the imaging diagnosis of 130 PLC cases with tumor thrombi in the portal vein was studied. Results 85%(110/130) cases of PLC with tumor thrombi in the portal vein belongs to type Ⅱ or Ⅲ when being first diagnosed in the hospital. The resectability rates for type Ⅰ, Ⅱ, Ⅲ, Ⅳ were 62%(5/8), 16%(10/62), 10%(5/48), 0 (0/12), respectively, and chemoembolization therapy (TACE) was given in 38%(3/8), 40%(25/62), 27%(13/48), 16%(2/12), respectively. Conclusions The new types of tumor thrombi are helpful for diagnosis and treatment of PLC with tumor thrombi in the portal vein.

Morphometric measurements of adenomatous hyperplasia of the liver by image analyzer / 中华消化杂志

Seven cellular parameters in adenomatous hyperplasia of the liver (AHL), compared with normal liver cells (NLC) and hepatocellular carcinoma (HCC), were analyzed quantitatively by measuring the seven parameters of the cell nuclei, the area (AREA), perimeter (PERIM), maximum diameter (DMAX), minimum diameter (DMIN), the equivalent circle diameter (DCIRCLE), circularity shape factor (FCIRCLE) and shape factor (FSHAPE) using computerized image analyzer. The main results were: (1) All the above seven nuclear parameters in AHL were significantly different from those in NLC, three parameters (DMAX, DMIN and FSHAPE) in AHL were not obviously different from those in HCC, indicating that AHL and NLC were not of the same cellular population, and AHL had a tendency of approaching HCC in the nuclear features; (2) Not alike NLC and HCC, the scattered distribution between nuclear FCIRCLE and DCIRCLE in AHL presented a highly dispersed distribution, overlapping the areas of NLC and the greater part of HCC, suggesting that AHL being an abnormal cellular population composed of cells in different proliferative status between NLC and HCC; (3) All of five resected AHL in the present study showed early malignant transformation. We conclude that AHL possesses the general features of precancerous lesion, and should be considered as the important precancerous lesion of HCC.

The relationship between gastroesophageal varices and bleeding and portal venous tumor thrombi in patients with hepatocellular carcinoma / 中国普通外科杂志

Objective To study the relationship between gastroesophageal varices and bleeding and portal venous tumor thrombi in patients with hepatocellular carcinoma(HCC). Methods From Jan. 2000 to Jan. 2003, 84 HCC patients with portal vein tumor thrombi were divided into Ⅰ～Ⅳ groups according to Ⅰ～Ⅳ types of tumor thrombi. The grade of gastroesophageal varices, the median survival time and the death cause for group Ⅰ～Ⅳ were retrospectively analyzed. Results Mild grade of gastroesophageal varices accounted for 64.7%, 6.0%, 85.7%,100%, respectively in group Ⅰ(n=17), Ⅱ(n=26),Ⅲ( n=35) and group Ⅳ(n=6),respectively. Severe varices were found in less than 5% in all four groups. The median survival periods were 10.1, 7.2, 5.7 and 3.0 months, respectively (P=0.0001). Most cases died from esophageal and gastric variceal bleeding and hepatic failure, with each accounting for about 50% of the mortality in all the 4 groups. Conclusions Portal venous tumor thrombi of HCC patients was not the major cause leading to esophageal and gastric varices and bleeding.

Observation of Hepatocellular Ultrastructure and G-6-Pase Cytochemistry in Hepatocarcinogenesis Induced by Diethylnitrosamine in Rats / 第二军医大学学报

In the process of hepatocarcinogenesis induced by diethylnitrosamine (DENA) in rats, the hepatocellular ultrastructure and G-6-Pase reactions in hepatic nodes were observed by electron microscope. The results are as follows: As compared with normal hepatocytes, cell junctions were fewer, even disappeared in some areas and intercellular spaces were wider; in some cells, nuclear membranes invaginated into the nucleoplasms frequently, micleoli were enlarged, mitochondria appeared swollen and their cristae were scanty- and short, and depolymerized ribosomes dropped off the dilated rough endoplasmic reticulums; in some seriously diseased cells, nucleoli were enlarged, abundant free ribosomes were present, but the' other organelles were in lower differencial state. G-6-Pase reactions were positive before the 8th week of DENA induction and negative after the 12th week. These suggest the hepatocellular metabolic disturbance and low differenciation.