1) Procedure for isolation of rat hepatocytes: About 30ml of 0.05% bacterial collagenase in Ca-free Hanks' solution was directly injected into the aseptically removed liver tissue through tuberculine needle at a speed of 20ml/min. After a few minutes of injection, liver tissue was torn into small pieces by two forcepts and then filtered through a platinum sieve with 150 meshes. Filtrate was centrifuged several times at a very low speed (200rpm). The final residue was suspended in Eagle's minimum essential medium containing 20% fetal calf serum and used for primary culture. The yield of isolated hepatocyte was (7.86±4.24)×106/g wet liver and contamination by Kupffer cells was less than 1%. More than 95% of cells excluded trypan blue.
2) Effect of prednisolone on gluconeogenesis and protein biosynthesis: Gluconeogenesis in cultured hepatocytes from alanine or lactate added to the culture media was enhanced by 100μg/dish of prednisolone. Contrary to it, incorporation of 14C-leucine or 3H-proline into the newly synthesized protein was inhibited by the same amount of prednisolone. Radioactivity of 3Hhydroxyproline was detected and the nature of the protein containing hydroxyproline was discussed.

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Hepatocellular transplantation into the spleen utilizing isolated hepatocytes has been investigated as a new attempt to compensate for impaired liver function. Present study was undertaken to evaluate ultrastructural alterations in hepatocytes transplanted into the splenic parenchyma up to 6 weeks following transplantation.
Light microscopical studies revealed viable hepatocellular islets in the splenic parenchyma, although minimal cytoplasmic changes were observed. Electron microscopical studies demonstrated moderate changes in organelles, which proceeded to develop gradually after transplantation. However, distortion and fragmentation of membranes of cytoplasm and organelles were not observed. Moreover, newly formed bile canaliculi and tight junctions which indicate reconstitution of hepatic trabeculae were observed between adjacent cell membranes.
The transplanted hepatocytes preserved their morphological characteristics with fine structures as hepatocytes up to 6 weeks. Our present sutdies suggest that inoculated hepatocytes maintain their hepatocellular functions after transplantation on the basis of the persistence of cellular viability.

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The possible role of liver cell plasma membrane (cell membrane) through the induction of hepatic injury by D-galactosamine (gal) was studied from the view points of 14C-amino acid pooling in the cell membrane and synthesis of the cell membrane protein. The specificity of the cell membrane which was collected by the method of Neville was examined by 5'-nucleotidase activity and electromicroscopic findings. The membrane protein synthesis was determined by the count of 14C(dpm) in each sections of gel, which were separated by means of 0.1% SDS-polyacrylamide gel electrophoresis. Amino acid pool and synthesis of the cell membrane protein decreased up to 2 hours after gal (400mg/kg body weight i.p.) injection. However, these findings mentioned above, was never found in the cell membrane from the rats which were treated with gal and uridine (1.2mg/g body weight i.p.).
Therefore, we want to point out the remarkable decrease of amino acid pool and synthesis of the protein in the cell membrane from gal treated rats within a short time when necrosis was never found. Then, the cell membrane may also play an important role for the induction of liver cell necrosis.

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The angiographic image of chronci active liver diseases supported the assumption for partial appearance of P-P or P-C bridge formation and some distortion of the lobular architecture by the presence of a bending and meandering of the hepatic artery at the terminus. In some cases, blood flow in the hepatic artery increased, and the bending and meandering of the artery extended to proximal vessels, accompanied by hypervascularity. This was considered to be related to the appearance of bright scarlet spots on the liver surface as seen by laparoscopy. Angiography would be highly useful for periodical observation of changes in the basic architecture of the liver diseases, especially in analyzing the formation and course of chronic acive liver diseases.

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Plasma LCAT activities were determined in patients with chronic liver diseases and in normal controls, by measurement of plasma cholesterol esterification during 2 hours' incubation with dipalmitoyl lecithin sols. In cirrhotics, plasma LCAT activities were reduced and the values were significantly correlated with the ratios of esterified cholesterol to total cholesterol (CE ratio) both in plasma and in HDL. The ratios of total cholesterol present in HDL to that in plasma were not significantly different between cirrhotics and controls. Highly significant correlation of LCAT activities to serum albumin levels, prothrombin time and ICG excretion were found in cirrhotics.
These findings support that in cirrhotics, decreases in both esterified cholesterol levels and CE ratios were mainly due to a reduction of plasma LCAT activities. Futhermore this reduction of LCAT activities was supposed to be related to a decrease in functional liver mass, which was demonstrated by the reduction of ICG excretion.

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Hemolytic complement levels, anticomplementary activity and cryoglobulin were examined in sera of 130 patients with various kinds of liver disease. Anticomplementary activity was found, in 39.0% of liver cirrhosis cases, in 28.6% of chronic hepatitis and in 5.6% of acute hepatitis but not in primary hepatoma nor fatty liver.
Anticomplementary activity was found frequently ill sera with complement levels lower than 10.0CH 50 (p<0.001) and also in sera with negative HBs antigen (p<0.05). A reverse correlation was observed between anticomplementary activity and complement level in liver cirrhosis (r=-0.576) and chronic hepatitis (r=-0.543).
Cryoglobulin was found in 9 cases of liver cirrhosis and 3 cases of chronic hepatitis. All of these 12 cases were negative for HBs antigen and 11 cases had anticomplementary activity.
These cryoglobulins were a IgG type in 7 cases and a mixed type of IgG and IgM in 5 cases, some of which contained Clq and/or RF. All samples of cryoglobulin had anticomplementary activity and their titers were higher than that of sera and anticomplementary activity of both cryoglobulin and serum was diminished after heating them at 56°C for 30 min.
Since the cryoglobulin deleted sera showed a marked decrease of anticomplementary activity, the cryoglobulin was considered as one of serum factors of anticomplementary activity.

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In this paper, Dane paticles and Hepatitis B core antigen were prepared from about ten liters of HBs Ag positive pooled plasma (subtype, adr. HBe Ag positive) And the polypeptide composition of purified HBc Ag was examined by SDS-polyacrylamide gel electrophoresis. Major polypeptide with approximately molecular weight 24, 000 was observed and variable polypeptides with molecular weight 17, 000, 37, 000, 46, 000 were present.

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A 73 year-old woman underwent surgery on December 4, 1975, because of leiomyoma of the small intestine and right adnexa. General anesthesia was performed by using G.O.F., and five units of whole blood were transfused during the operation. The operation was successfully done within two hours. On the first postoperative day jaundice developed without pruritus or hepatosplcnomegaly.
The serum bilirubin rose to 9.7mg, the conjugated bilirubin level was 6.5mg per 100ml. Serum alkaline phosphatase level elevated also. However, through the icteric period, the elevations of serum glutamic pyluvic and oxaloacetic transaminases remained within 313mU. and 113mU. per ml, respectively. A needle liver biopsy, performed on the 17th postoperative day, showed a picture of pure intrahepatic cholestasis; i.e., bile thrombi in the canaliculi and bile pigments in the cytoplasms of hepatocytes were recognized, in spite of no evidence of inflammation and liver cell necrosis in the portal tracts and lobules. Up to the 20th postoperative day, the jaundice disappeared completely, and serological tests returned to normal values also.
The clinical course and laboratory findings of this case were considered to be compatible with those of benign postoperative intrahepatic cholestasis, which was described by Schemid and others.

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A case of lupoid hepatitis with portal hypertension was reported. This patient was 23-year old woman, She was admitted to a hospital with complaining of jaudice, high fever and ascites about 3 years ago. Laboratory examination in the hospital revealed abnormalities of liver function and positive L.E. cell phenomenon. She was treated with predonisone on suspicious of SLE for 2.5 years. There was an improvement of these symptoms but suddenly, hematoemesis from esophageal varices occured.
Transabdominal esophageal transection with devascularization of esophago-gastric region and splenectomy was performed because of reccurent bleeding from esophageal varices. During operation, the spleen was markedly enlarged and weighed 1, 440g and liver cirrhosis with multiple nodules was observed. Portal pressure was 400mm H₂O.
Microscopic examination demonstrated coarsely nodular pseudolobules and marked infiltration with lymphocytes and plasma cells in the liver and numerous prominant lymphoid follicle in the spleen. By orcein staining, HBs antigen of the liver specimens could not be found.
Preoperative laboratory data inculuded, hemoglobin, 7.9g/100ml, WBC, 1, 700/mm3 and platelet count, 33, 000/mm3. The total serum protein was 6.8g/100ml with albumin 2.2g/100ml and gamma globulin 3.5g/100ml. Immunological studies showed high value of IgG and low level of C₃, C₄ (third and fourth components of the complement system). The latex agglutination antinuclear antibody and antismoothmuscle antibody reaction was positive but the L.E. cell phenomenon and antimitochondrial antibody reaction was negative. Moreover, a decreased B-cell population and reduced PHA responsiveness of peripheral lymphocytes was observed. Many serological and immunological abnormalities were noticed in this patient. From these results, lupoid hepatitis with portal hypertension was diagnosed.
It has been reported to explain that portal hypertension with splenomegaly in this disease is due to obstruction of portal venous blood flow. However, it is suggested that immunological disorders may be related to portal hypertension with splenomegaly.

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A 57 year-old male docter was found to have a large abdominal tumor in the right upper quadrant. 1⁹⁸Au colloid liver scintigram revealed multiple defects scattered in the right lobe. Serum A.F.P. was 23ng/ml. Two years later an exploratory laparotomy was performed. Tumors were sharply circumscribed and separated from the hepatic parenchyma by tight fibrous capsule.
Microscopically, the tumor was well differenciated type of hepatocellular carcinoma-Edomondson's grade I. Such cases of hepatocellular carcinoma which had long-term survival more than 2 years were reviewed, and discussed about the term of "slow growing hepatoma."

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A case of remakable obstructive jaundice caused by hepatocellular carcinoma was described.
A 77 year-old male was admitted to our hospital with symptoms of cholelithiasis. The patient was diagnosed as hepatocellular carcinoma because of high serum a-fetoprotein value and angiographic findings. As jaundice was progressive in the clinical course, percutaneous transhepatic cholangiography performed that revealed two obstructive areas in the bile duct. One was at the joint of the right and left hepatic ducts and another was at the duodenal end of the common duct.
The autopsy revealed that hepatocellular carcinoma infiltrated in the bile duct and obstructed the extrahepatic bile duct, but no mass was seen at the duodenal end. By pathological examination, it was suspected that lower obstructed area revealed by percutaneous transhepatic cholangiogram was due to the necrotic mass of the carcinoma.

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