Each part of C gene, S gene and X gene of HBV DNA was amplified by polymerase chain reaction (PCR) to 10⁷, 10⁷ and 10⁶, respectively. Using this PCR system, we examined whether HBV DNA sequence was detected in formalin-fixed paraffin-embedded sections of liver specimens. Not only eight cases with free form of HBV DNA but also nine cases with only an integrated form of HBV DNA were positive for HBV DNA sequence in spot hybridization and Southern blot analysis of PCR products. Thus PCR was useful to detect HBV DNA in a thin section of formalin-fixed paraffin-embedded liver specimens.

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The expression of c-myc and c-Ha-ras products were investigated in chronic liver disease by indirect immunohistochemical method. The c-myc product was observed in 29.8% (14/47) patients with chronic hepatitis and in 27.3% (3/11) patients with liver cirrhosis. The c-Ha-ras product was observed in 19.1% (9/47) patients with chronic hepatitis and in 18.2% (2/11) patients with liver cirrhosis.
Among HBV carriers, the expression of X-protein and HBcAg was observed in successive sections of liver tissues. The c-myc product was more often observed in tissues with X-protein than without X-protein. Also, the c-Ha-ras product was more often observed in liver tissues with X-protein and HBcAg. The positive frequency of HBeAg and the values of DNA polymerase and a-fetoprotein were higher in the c-myc or c-Ha-ras positive patients than in the negative.
These results suggest that the expression of c-myc and c-Ha-ras products may be induced as a signal of liver regeneration after cell damage by the active replication of HBV.

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Two cynomolgus monkeys were intravenously inoculated with a stool extract derived from Burmese patients who suffered from hepatitis E. The monkeys developed self-limiting acute hepatitis. Liver tissues were obtained sequentially by needle biopsy or at sacrifice. Histologically, the liver tissues exhibited necroinflammation which was characterized hy focal dropout of lymphocytes with accumulation of lymphocytes and macrophages. The severity of necroinflammation appeared to be parallel with elevation in serum aminotransferases. The lymphocytes in necroinflammation were positive for a cytotoxic/suppresser T (Leu-2a) immunophenotype. The hepatocytes surrounding focal necroses showed depletion of glycogen granules and decrease in glucose-6-phosphatase and succinic dehydrogenase activities. Ultrastructurally, damaged hepatocytes around the focal necroses revealed marked injurious change of the cytoplasmic organelles. Virus-like particles measuring approximately 27nm in diameter were observed in the hyaloplasm of damaged hepatocytes which frequently contacted with lymphocytes. These findings strongly suggested the immune-mediated hepatocytolysis in hepatitis E.

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When the heat-killed Propionibacterium acnes (P. acnes) was injected into mouse through tail vein, macrophages including Kupffer cells accumulate in the liver reaching its maximum 7 days later. To study the function of these liver adherent cells, production of tumor necrosis factor (TNF) and its regulation by prostaglandins (PGs) were investigated in BALB/c mouse and C3H/HeJ mouse. As a result, in BALB/c mouse, P. acnes-elicited liver adherent cells and Kupffer cells produced TNF by stimulating with endotoxin lipopolysaccharide in dose-dependent manner, but in C3H/HeJ mouse, not. When, in BALB/c mouse, these liver adherent cells and Kupffer cells were treated with PGE and PGI₂, production of TNF was significantlly reduced. These results suggested that TNF may play a role in the induction of acute hepatic cell necrosis model, and inhibition of TNF production by liver adherent cells and Kupffer cells possibly suppress liver injury in this model.

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When mouse hepatic sinusoidal endothelium was stimulated with lipopolysaccharide (LPS), interleukin-1 (IL-1) was synthesized from hepatic sinusoidal endothelium. This IL-1 synthesis decreased from hepatic sinusoidal endothelium stimulated with LPS and prostaglandin (PG) E₁ and E₂, but increased significantly from hepatic sinusoidal endothelium stimulated with leukotriene B₄ or platelet-activating factor (PAF).
These results was suggested that IL-1 synthesis from hepatic sinusoidal endothelium may be regulated by arachidonic acid cascade and PAF.

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The amounts of prothrombin-related antigen, des-gamma-carboxy prothrombin and vitamin K concentration were determined in human liver tissues. Both prothrombin-related antigen and desgamma-carboxy prothrombin contents were significantly increased in hepatoma tissues as compared with non-cancerous liver tissues. Tissue des-gamma-carboxy prothrombin content was well correlatedwith plasma levels in patients with hepatocellular carcinoma, suggesting a tumor production of desgamma-carboxy prothrombin. Between the amount of prothrombin-related antigen, however, did not correlated neither with tissue nor plasma levels of des-gamma-carboxy prothrombin. Prothrombin synthesized in hu-H1 cells eliminated rapidly, which was demonstrated by pulselabeling using 35Smethionine, suggesting that hepatoma cells have no defect in its ability to excrete prothrombin. So it was indicated that over-production of prothrombin (prothrombin precursor) may play some role in the synthesis of des-gamma-carboxy prothrombin. Vitamin K (K1, MK-4, MK-7) concentration revealed no significant difference between in cancerous and non-cancerous portion of the liver, indicating that the significant difference between in cancerous and non-cancerous portion of the liver, indicating that the possible vitamin K deficiency is not essential.

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The changes in cytokeratin (CK) intermediate filaments (IFs) of the hepatocytes in the hyperplastic nodules (HNs) of the rat's liver developed by Solt and Farber's model were studied by light and electron microscopy, immunofluorescence (IMF), immunoelectron microscopy (IEM) and immunoblotting (IMB). Frozen sections of liver tissues were extracted by Triton X-100 and nuclease. IEM was performed by using monoclonal anti-CK antibody (PKK1: CK55 and CK49). CK staining by IMF was markedly decreased in the hepatocyts in HNs compared with that in the parenchyma surrounding HNs; however, IEM clearly showed a lot of non-staining IFs in hepatocytes in HNs. In the IMB, two bands in insoluble proteins extracted from control rat liver and the experimental rat liver including HNs reacted with CK55 and CK49, respectively. These findings indicate that immunocytochemical change of IFs of hepatocytes might be occurred in the precancerous lesion of the experimental hepatocarcinogenesis in rats.

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Studies were conducted to investigate the metabolism of monoamines, in particular that of norepinephrine (NE), in the brain of rats with hepatic failure. Hepatic failure was induced in rats by either acute ischemic hepatic failure (AIHF) or portacaval anastomosis (PCS). Brain levels of NE and its metabolites including 3-methoxy-4-hydroxyphenylglycol (MHPG), the final metabolite of NE in the central nervous system, were determined by HPLC-voltammetry.
The following results were obtained:
1) AIHF rats showed significantly higher total-MHPG and lower NE levels in the brain as compared with controls.
2) In AIHF rats, a significant negative correlation between NE and either phenylalanine (Phe) or tyrosine (Tyr), as well as a significant positive correlation between MHPG and Phe or Tyr were demonstrated in the brain.
3) Altered NE and MHPG levels in the brain of AIHF rats were found to restore toward normal following the treatment with Fischer's solution.
4) PCS rats showed significantly higher total-MHPG levels in the brain as compared with controls, while no significant change was observed in NE level.
In conclution, the reduction of NE level in the brain of AIHF model appears to be caused, at least in part, by the enhanced catabolism of NE due to increased brain contents of Tyr and Phe. This finding seems to support the Fischer's hypothesis of "false neurotransmitter" in hepatic encephalopathy.

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Proper hepatic artery diameter (PH) and indocyanine green cirucration time from common hepatic artery to ear (HA-E time) were measured to see the changes in hepatic artery hemodynamics in patients with hepatocellular carcinoma.
PH did not change with the size of tumor, but HA-E time was shorter when the tumor occupied more than 60% of the area of the liver in a film taken at angiographic study. PH was not influenced by the existance or site of portal thrombosis (V), while HA-E time was also shorter when V was seen in the trunk than when it was seen in the lst order branch of portal vein. HA-E time was also shorter when arterio-portal vein (A-P) shunt which had retrograde flow was seen at the lst order branch of portal vein, than when A-P shunt was seen in the periphery.

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Ozawa et al showed arterial blood ketone body ratio (acetoacetate/β-hydroxybutyrate, AKBR) reflects the mitachandrial free NAD+/NADH ratio which is coupled with mitochondrial oxidative phosphorylation. In the study, changes in the AKBR during 27 hepatectomies were examined in relation to major post-operative complications involving hepatic failure. Concentrations of arterial blood ketone bodies, acetoacetate and β-hydroxybutyrate, were determined at least three times during the operation, i.e, at laparotomy (phase I), during hepatectomy (phase II), and immediately after operation (phase III), according to the standard method reported by Williamson. AKBR of all 22 patients varied from minimal level 0.22 to maximal 10.8 in phase I, decreased with the progress of procedures, and returned to normal range (over 0.7) in phase III except in 5 cases. Four patients out of the 5 patients with low levels of AKBR in phase III died of severe hepatic failure. The other cases whose AKBR returned to normal range in phase III did not show any complications. The measurement of AKBR during partial hepatectmy is of major importance in the prognostic evaluation of a postoperative risk.

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Nodular lesions in the liver with idiopathic portal hypertension (IPH) were observed in 14 out of 40 autopsy cases (35%), and in 4 out of 51 surgical cases (7.8%). We examined the following 11 cases, classifying into two groups; portal hypertensive group (8 cases) and non-portal hypertensive group (3 cases). 4 cases with IPH prominent nodular lesions, 6 cases with nodular regenerative hyperplasia (NRH) and 1 case with partial nodular transformation (PNT). The results were as follows; 1. Portal hypertensive group showed the decreased liver weight, the increased spleen weight and the histological findings including scarring of the peripheral portal vein, portal sclerosis and abnormal pathway.
As to the relationship between portal hypertension and the nodule (hyperplasia of the hepatocytes), it was likely that the nodule might secondarily develop in association with the pathogenesis which lead to portal hypertension. The following reasons were cited; 1. Multiple nodular transformations were found in NRH without portal hypertension. 2. The nodular formation of various degree was found in IPH. 3. The collapse of the peripheral portal vein was also found in NRH and PNT.

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We studied the abnormality of extrahepatic portal hemodynamics in 37 cases with liver cirrhosis, by percutaneous transhepatic portography and divided the subjects into two groups, i.e. 20 cases having mainly the collaterals except esophageal varices [group I]and 17 cases of the esophageal varices [group II].
In group I, there were 8 paraumbilical, 4 gastro-renal, 2 spleno-renal and 6 superior or inferior mesenteric vein-inferior vena cava shuntings. Twelve cases (60%) had hepatic encephalopathy in group I (5 paraumbilical, 1 spleno-renal and all 6 superior or inferior mesenteric vein-inferior vena cava shuntings). On the other hand, only one case (6%) had hepatic encephalopathy in group II, whose esophageal varices were diverted from the superior mesenteric vein. The occurrence of hepatic encephalopathy had a statistical significance between the two groups (p<0.005).
The values of ICG R15 were 47.4±12.7% in group I and 33.2±11.0% in group II(p<0.01).In addition, a significant negative correlation between ICG R15 and portal pressure was observed in gorup I, but not in group II.
In conclusion, depending on the main collaterals, the clinical manifestations of liver cirrhosis can be varied.

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Idiopathic portal hypertension (IPH) is characterized pathologically by portal fibrosis and phlebosclerosis, but the etiology of IPH has not been elucidated yet. To clarify the etiology, we attempted to make histopathologic findings in the liver similar to those of IPH by intraportal injections of endotoxin (LPS).
Five μg of LPS was injected into the portal vein as the preparation of Schwarzman phenomenon in the rabbits of group I with gelform powder (Gel) and group II without Gel, and the same amount of LPS was injected into the ear vein as the provocation at 24hr after the preparative injection. In group III, 50 μg of LPS was injected into the portal vein at 1-week interval a total of four times to elucidate the effects of long-term and repeated injection of LPS.
Liver histology showed stenosis and collapse of peripheral portal tracts and aberrant vasculature in portal areas. In group III, dense portal fibrosis was recognized.
These changes closely mimic those seen in human IPH, and it was suggested that the long-term and repeated inflow of LPS to the liver via mesenteric veins may play a role in the development of IPH.

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A case of chronic hepatitis B with HTLV-I healthy carrier which showed Type I response (Merigan) to Adenine Arabinoside (Ara-A) therapy was reported. A 20 year-old-man was diagnosed as chronic active hepatitis by needle liver biopsy. Before Ara-A therapy he was HBV carrier with HBe antigen positive. Just after Ara-A was given, his serum aminotranseferase level began to fluctuate remarkably and was followed by the disappearance of HBeAg and HBsAg 2 months and 5 months later respectably, and appearance of anti-HBs was noted 7 months later. The fact that no familial clustering of HBV but of HTLV-I indicated that he became HBV carrier by horizontal transmisison and HLTV-I carrier by vertical transmission. This is the first report of Type I response induced by Ara-A therapy alone, and is thought to be a valuable case to support the hypothesis that HBV carrier state by horizontal transmissionis terminates more easily than that by vertical transmission.

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A 55-year-old woman was found to have mass in the right lobe of the liver by computed tomography and ultrasonography. Hepatic arteriography showed that the mass was hypovascular. Tumor biopsy showed most of the specimen AH but small portion of it was well-differentiated hepatocellular carcinoma. Six moths later, AFP increased and echo pattern of the main mass was changed and furthermore we detected another masses. Hypervascular mass appeared in part of the main mass by appeared in part of the main mass by arteriography, and then TAE and PEIT were performed. After the treatment AFP decreased but six months later AFP increased again. Tumor biopsy of the main mass was diagnosed as Edmondson type I hepatocellular carcinoma, which was more undifferentiated than the earlier specimen. This case suggests that borderline lesion, which is difficult to diagnose as AH or hepatocellular carcinoma, may soon or later progress to obvious hepatocellular carcinoma, and so we must carefully follow up such cases.

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