Fifteen adult patients (21-60y.) with AML (12 fresh and 3 relapsed) who had achieved complete remission, were treated with AcDAc regimen as an intensive consolidation therapy. Their clinical courses were followed without further chemotherapy (Group A). The results were compared with those obtained in 17 adult patients (26-60y.) who were treated with2 or 3 courses of BH-AC DMP regimen as a standard consolidation therapy and thereafter with 6MP/ara-C as a maintenance therapy following achievement of complete remission (Group B) to determine the efficacy of intensive consolidation therapy in improving the prognosis. Rates of surviving patients and of patients in complete remission were always higher in Group A than Group B, although the differences were not statistically significant. Myelosuppression after consolidation therapy was more severe in Group A than Group B judging from the changes in peripheral blood and bone marrow findings. This was confirmed by the evaluation of changes in CFU-GM of the bone marrow. All consolidation courses in Group A were complicated by severe infection and 3 patients were died of fulminant hepatitis or pneumonia, while only one patient was complicated by infection and all were survived in progress of consolidation therapy in Group B. In Group A, more than 70% of patients had gastrointestinal toxity and eruption. These were mild and well tolerated. It is concluded that intensive consolidation therapy like AcDAc regimen given in aseptic environment seems to be superior to standard consolidation therapy with maintenance therapy for long time in the treatment of adult patients with AML.

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Attempts were made to develop a metabolic model for human megaloblastic anemia due to cobalamin (vitamin B₁₂) or folate deficiency using a cell culture system. When cultured in the folate-and B₁₂-deprived medium supplemented with CH₃-THF at 10 ng/ml or 250 ng/ml, the L1210 cells were rendered folate-deficient 48 hours after transfer. This was evidenced by retarded growth, decreased DNA synthesis and marked improvement of deoxyuridine (UdR) incorporation following either folic acid or CH₃-THF addition, whereas B₁₂ did not significantly affect these parameters. However, pretreatment of the medium with nitrous oxide resulted in the development of B₁₂-deficient state, as indicated by improved UdR incorporation following B₁₂ addition. The results also indicated that, at least in this culture system, a concomitant folate-deficiency was a prerequisite for the development of overt B₁₂-deficiency by nitrous oxide exposure. This model system may be useful as an experimental tool for further elucidation of the biochemical mechanisms of megaloblastic anemia.

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Primary biliary cirrhosis (PBC) is histologically characterized by chronic non-suppurative destructive cholangitis (CNSDC), and is also a granulomatous disease. The characteristics of granulomas and the correlation between granulomas and bile duct changes were morphologically investigated in 31 patients with granulomas, compared with in those without granulomas.
Results were as follows :
1) Most of the granulomas (93%) were poorly-defined, and well-organized granulomas with distinct fibrosis were rare (7 %).
2) Granulomas with multinucleated giant cells (MGCs) were not always well-organized and were found both in portal area and near central veins. MGCs sometimes had inclusions with negative H-E stain.
3) Seventy-two per cent of granulomas were found in portal area, some of them were in contact with damaged bile ducts, others surrounded them.
4) The patients with granulomas were rather early in the histologic stages. Twenty-three of 31 patients with granulomas were asymptomatic.
5) Ratio of disappearance of interlobular bile ducts was significantly lower in PBC patients with granulomas than in those without granulomas.
From these results, it was suggested that granulomas in PBC were closely related to interlobular bile duct lesions and that something derived from interlobular bile ducts seemed to play an important role for granuloma formation.

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Ultrastructural changes of intrahepatic bile ducts in primary biliary cirrhosis (14 cases) and chronic hepatitis (9 cases) were investigated.
Results were as follows :
1) Disruption of basement membrane and falling of epithelial cells into periductal space were revealed in a large bile duct and a medium-sized interlobular bile duct which showed chronic non-suppurative destructive cholangitis (CNSDC), and in a large bile duct without CNSDC in PBC.
2) Infiltration of lymphocytes into bile duct epithelium was more prominent in PBC (12 of 27 bile ducts) than in chronic hepatitis (4 of 22 bile ducts).
3) In PBC, “dark cell” metamorphosis was often observed in the epithelial cells in contact with the lymphocytes infiltrating into bile duct epithelium.
From these results, in PBC, disruption of basement membrane and falling of epithelial cells into periductal space may be specific changes in process of disintegration of bile ducts and sensitized T lymphocytes seemed to be responsible for the bile duct damages.

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In order to study the nutritional assessment after total gastrectomy, ¹³¹I-triolein and vitamin B₁₂ absorption tests were performed and other parameters were measured over ten years from the viewpoints of reconstruction procedures performed, aging of the patients and pathological stage of gastric cancer. The following results were ascertained.
1) A significant hindrance in fat and vitamin B₁₂ absorption was occurred after total gastrectomy.
2) Reconstruction method after total gastrectomy and aging of the patients affect upon the nutritional status after the surgery.
3) The recovery of each chemical and/or physical parameter was delayed considerably and not recover to the preoperative level after the operation.
4) Postoperative malnutrition is observed immediately after the operation. In such condition, measurement of rapid turnover transport protein appears to be useful for the sensitive determination.
5) In order to improve subclinical malnutrition after total gastrectomy, powerful and positive countermeasures such as total parenteral nutrition and enteral nutrition should be applied.

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Immunochemical techniques were applied to the investigation of the in vivo distribution of the Parotin subunit, extracted from bovine parotid glands, which decreases serum calcium. The results were as follows.
1) Using as an antigen the bioactive Parotin subunit, purified and isolated from bovine parotid glands, specific antibodies to this antigen were produced.
2) Using a radioimmunoassay method, these antibodies were found to have a high titer, and in cross-reactivity and Ouchterlony tests as well as immunoelectrophoresis, were shown to be specific for the Parotin subunit.
3) Using the PAP enzyme-antibody staining technique, strong staining reactions to these antibodies were found also on the duct cell of bovine, guinea pig and human parotid and submandibular glands as well as sublingual and minor salivary glands. In addition, similarities were observed in pancreatic and gastric glandular and mucosal epithelium. Moreover, staining reactions to these antibodies were also detected in the tumor cells of the duct cell or the hyperkeratic epithelial cell on pleomorphic adenoma adenocarcinoma etc. of human salivary glands. Serum Parotin subunit labeled with ¹²⁵I was distributed in liver and kidney of guinea pig.

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Urinary hydroxyproline (HP) excretion was estimated, without prior dietary restriction in 70 patients (52 patients with prostatic cancer and 18 patients with benign prostatic hypertrophy) and expressed as either 24hr-HP output or as the hydroxyproline/creatinine (HP/Cr) ratio in a 24hr urine sample, an early morning urine sample, and spot urine sample.
Although each of the three different samplings had a wide daily variation in patients with active or controlled bone metastases, their HP and HP/Cr values were significantly higher than those in patients with localized prostatic cancer and benign prostatic hypertrophy. Among the three sampling types, 24hr-HP/Cr appears to be the most valuable for diagnosis.
The HP/Cr average over three days is useful in monitoring the response to treatment in patients with bony metastatic cancer. Among HP/Cr, Prostatic acid phosphatase and Alkaline phosphatase, HP/ Cr and Alkaline phosphatase are more sensitive markers of bony metastases.

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The withdrawal effect of spironolactone treatment on natriuresis was studied in relation to atrial natriuretic peptide (ANP) and digitalis-like substance (DLS) in 5 patients with primary aldosteronism due to adrenal adenoma. The patients had been treated with spironolactone for 2-3 months before they were admitted. After admission, blood pressure, body weight and urinary excretion of DLS and sodium were measured daily. Venous sample were obtained twice a week for measurements of plasma renin activity (PRA) and plasma levels of aldosterone, ANP and DLS. Plasma volume was determined by radioisotope dilution using ¹²⁵I-human serum albumin during the control period and on the 13th day after stopping spironolactone. The study was performed for 7 days during the treatment with spironolactone and for 18 days after stopping the administration. Urinary sodium excretion decreased initially and returned to the control levels successively. Body weight and plasma volume increased, and blood pressure rose steadily. PRA decreased markedly (p <0.05) from 2.1 ± 0.6 ng/ml/hr on the control days to 0.2 ± 0.1 ng/ml/hr on the 8 th day after stopping spironolactone. Plasma levels of aldosterone was 387± 150 pg/ml on the control days and did not change significantly after the withdrawal (278 ± 94 pg/ml on the 17th day). Plasma ANP levels increased significantly (p<0.05) from 26 ± 4 pg/ml on the control days to 195 ± 47 pg/ml on the 13th day ; however, plasma and urine levels of DLS decreased significantly (p <0.05). The data of the urinary sodium excretion showed the escape from sodium-retaining effect of aldosterone, and this escape could be explained by the increase in plasma ANP but not by the changes in DLS.

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A physiological role of somatostatin (SRIF) on GH-releasing factor (GRF) -induced GH secretion was examined by utilizing in vitro perifusion systems. The following three experiments were carried out.
1. GRF-induced GH release was examined in whole anterior pituitaries from rats subjected to the following treatments : 1) anterolateral hypothalamic (ALH) deafferentation, 2) lesions of the hypothalamic periventricular nucleus (Pe) and 3) passive immunization with anti-SRIF serum. The response of these pituitaries to human (h) GRF (0.1 μM) stimulation was markedly attenuated within a day, accompanied by a significant decrease in SRIF content in the stalk-median eminence from rats with ALH deafferentation and Pe lesions. Also SRIF in the SME significantly decreased after the ALH cut or Pe lesions, whereas hGRF remained constant even 10 days after the operations. These data suggest that SRIF in vivo plays a significant role in the anterior pituitary in maintaining hGRF-GH response in vitro.
2. The effects of SRIF infusion on successive GH release in response to GRF were examined in an in vitro system as in Exp. 1. Two hours after the first GRF (0.1 μM) stimulation, the response to the second stimulation was markedly attenuated. But this was not observed when the pituitaries were subjected to the second stimulation after a 3 hour interval. In contrast, when SRIF (0.1 μM) was infused for 50 min 1 h after the first stimulation to lower basal GH release, the second response to hGRF was restored to the level of the first response. To determine whether SRIF exerts a direct action on the GH response, a prestimulatory perifusion with SRIF (0.1 μM) for 50 min was tested. The treatment tended to facilitate the pituitary response to hGRF. When pretreated with SRIF for 50 min at a lower concentration (0.05 μM), the response to the first hGRF (0.05 μM) stimulation was significantly facilitated. These data suggest that 1) SRIF perifusion rapidly restores the attenuated response to a second hGRF challenge by lowering GH release to basal levels and 2) SRIF pretreatment facilitates the GH response to the first hGRF stimulation.
3. The facilitation by SRIF of hGRF-induced GH response of dispersed pituitary cells was further characterized and analyzed. SRIF-pretreatment (1.0 nM) also caused a marked facilitation of the hGRF (1.0 nM) -GH response which continued for 30 min after SRIF withdrawal. Although a significant “rebound increase” in GH release was observed within 15 min after SRIF withdrawal, the amount was much less than in the hGRF-GH response. Both DbcAMP (1.0 mM) -and KCl (15 mM) -induced GH release response were significantly facilitated by SRIF pretreatment. However, cAMP production by hGRF was not increased but significantly decreased by SRIF pretreatment. Thus SRIF facilitation of hGRF-GH release is thought to take place in the process after cAMP formation.
From the results of these experiments, it is concluded that 1) SRIF is required for the maintenance of the pituitary response to GRF, 2) SRIF pretreatment facilitates GRF-induced GH release.

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Three continuous cell lines (GMC-N, GMC-U GMC-Y) have been established from different primary breast cancers. To remove contaminated fibroblasts from epithelial cells, floating cells in the primary culture were serially transferred to a new dish and the resultant cells were cultured in serum-free medium with 8 supplements.
GMC-N and GMC-U cells were polygonal and typical epithelial cells, but GMC-Y cells were smaller and rounder than GMC-N or GMC-U cells and tended to pile up.
The doubling times of GMC-N, U, Y cells were 18.3, 41.9, 18.2 hr, respectively.
Estrogen receptor (ER) and progesterone receptor (PgR) were measured by whole cell uptake method. Both GMC-N and GMC-U cells were ER-and PgR-positive, while GMC-Y cells were ER-positive, but PgR-negative. Estradiol-17β stimulated the growth of all these cell lines at 10^-10M concentration.
All of three cell lines were tumorigenic to athymic nude mice and solid tumors developed 3 weeks after subcutaneous injection in all mice. Histological examination of the grafted tumors revealed the similar morphology with minute cavities in all cases.
These results indicate that these three established cell lines could be useful in studying the biological properties of breast carcinoma cells and the mechanisms of drugs used in endocrine therapy.

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Eighty nine healthy volunteers working at medical institution were given inactivated HB vaccine and followed up changes of HBs-Ab levels for 49 months.
Three doses of vaccine induced antibody in 70.5% out of 78 vaccine recipients at 28 weeks after the first dose. Active immunization occurred more frequently in female than in male.
At 49 months after the first dose, HBs-Ab positive rate decreased to 37.4% out of 32 subjects. All of those positive volunteers have had high HBs titer (Cut off index≥50) at 7 months after the first dose. But there was no sex or age difference in HBs-Ab titer.
These results suggest that inactivated HB vaccine is useful for active immunization of HBs-Ab. However, further investigation is needed to make clear whether re-vaccination is necessary or not.

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A retrospective clinical analysis was carried out on 39 patients with neuroblastoma treated from 1960 to June, 1988. Survival rate was analyzed according to stage, primary site, histology and age. Two-year survival was as follows : stage I, II and IVS, 100% ; stage III, 53% ; stage IVB, 40% : and stage IVA, 0 %. For all stages, patients younger than 1 year of age survived longer than those older than 1 year of age (p <0.05). However, prognosis for stage IV (stage IV S is not included) was poor regardless of age. The primary site and histology were not important with respect to prognosis. For advanced neuroblastoma (stage III and IV) the survival rate tends to be improved after 1984, but is not significant. From these results it is concluded that management of neuroblastoma should be altered to minimize therapeutic strategy in stage I, II, III (patients younger than 1 year of age) and IVS, and to improve survival in stage III (patients older than 1 year of age) and IV.

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The effects of insulin treatment on body fluid volumes and plasma renin activity were studied in 7 normotensive non-ketotic diabetic patients. Blood and urine samples were obtained for measurements of fasting blood glucose, daily excretion of urinary glucose, hematocrit, plasma renin activity and plasma aldosterone concentration on the control days of poor metabolic control and twice a week after starting the treatment with insulin. The dosage of insulin was increased by the levels of fasting blood glucose. Body fluid volumes were also measured on the control days and on the 14th day after starting the treatment ; plasma volume and extracellular fluid volume were determined by dilution of Evans blue and sodium thiocyanate, respectively.
After starting the treatment, hematocrit decreased and both plasma volume and extracellular fluid volume increased significantly, accompanied by a fall in fasting blood glucose and daily excretion of urinary glucose. Plasma renin activity decreased significantly from 3.9 = 1.6 ng/ ml/hr on the control days to 0.7 ± 0.3 ng/ml/hr on the 14th day ; however, plasma aldosterone concentration did not change. The increase in body fluid volume may be attributable to a sodium-retaining effect of insulin on the kidney and a decrease in osmotic diuresis of urinary glucose. The increase in body fluid volumes may contribute to a fall in plasma renin activity. Further studies are needed to elucidate the reason why plasma aldosterone concentration did not alter.

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Plasma lipid, lipoproteins, apolipoproteins and lecithin : cholesterol acyltransferase (LCAT) activity were measured in 56 non-insulin-dependent-diabetic patients (34 males and 22 females) before and after the admission of approximately 15 days. The treatment was sulfonylurea administration in addition to diet and exercise.
Compared with the pre-admission, the following results were obtained at the time of discharge,
1) bodyweight, fasting blood glucose, triglyceride and total cholesterol were significantly decreased. In addition, high-density-lipoprotein (HDL) -cholesterol significantly decreased from 47 ± 15mg/dl to 44±13mg/dl (P < 0.01),
2) apolipoproteins (A-I, A-II, B, C-II, C-III, E) were significantly decreased, however, apolipoprotein A-I /A-II ratio did not change,
3) HDL3-cholesterol was significantly decreased from 27 ±7 mg/dl to 22 ±5 mg/dl (P < 0.05), however HDL2-cholesterol did not change,
4) LCAT activity did not change during the hospitalization.
It is not clear whether such a significant decrease of HDL-cholesterol and HDL3-cholesterol in the patients with sulfonylurea treatment has any causative relationships to the development of atherosclerotic vascular disorders.

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