Heparin has been widely used as a potent anticoagulant for several decades and the mechanism of the action has almost been fully elucidated by recent aggressive biochemical investigation. The mode of administration of heparin, however, has not fully established yet, although various methods has been advocated such as bolus intravenous (i. v. bolus), continuous intravenous (i. v. cont.) or subcutaneous injection.
In the present study, the anticoagulant effect of heparin in rats was monitored and compared, employing different modes of administration. It was possible to maintain constant hypocoagulability by a combination of i. v. bolus and i. v. cont. but dosing was extremely difficult. A subcutaneous administration produced constant hypocoagulability of relatively longer duration compared with i. v. bolus or i. v. cont. Also, inhibitory effect on disseminated intravascular coagulation (DIC) in rats was studied with different methods of administratlon. The inhibition by heparin on DIC in rats was totally dependent upon the degree of hypocoagulability before DIC was triggered, regardless of the mode of administratlon. Clinically, subcutaneous administration of heparin was applied in 8 cases for the prevention and treatment of DIC and in 6 cases for those of thromboembolic diseases. The clinical dose of subcutaneous heparin was too small to be detected by conventional coagulation tests but overall clinical results were favorable, especially for the prevention of DIC, though number of patients was very limited. More extensive clinical trials are required to confirm the efficacy of subcutaneous heparin for the prevention of DIC but subcutaneous administration of heparin is also promising in other cases requiring mild hypocoagulability than in well documented effect in the prevention of postoperative deep vein thrombosis or pulmonary embolism.

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Serum ferritin level was determined in 187 normal subjects of various ages and in various gynecological diseases with special attention to uterine cancer.
(1) Logarithmic mean values (±2 standard deviation) in normal subjects were as follows: women at 20 to 49 years old, 20 (5∼80) ng/ml: women at 50 to 59 years old, 30 (6∼145) ng/ml: women at 60 to 75 years old, 48 (13∼182) ng/ml: men at 20 to 35 years old, 63 (16∼252) ng/ml. These results indicate that, for evaluating on serum ferritin in gynecological diseases, age and sex variations in serum ferritin must be considered.
(2) In 26 patients with myoma uteri accompanied with hypermenorrhea, the decrease of serum ferritin was observed. In 9 patients with primary amenorrhea, serum ferritin values were higher than in normal women at the same age. From these results, the sex difference of serum ferritin may be attributable to the menstruation. However, serum ferritin values in 9 patients with primary amenorrhea and in 27 post-menopausal women were still lower than in normal adult men, so that the sex and women's age difference of serum ferritin can not be attributed to the menstruation alone.
(3) In 25 patients with untreated uterine cancer at 0 to III stage, serum ferritin was within normal limits. In 2 out of 3 patients with untreated uterine cancer at IV stage, high serum ferritin level was observed. However, hyperferritinemia found in these two patients was associated with low TIBC, SI and Hb, indicating that it appears to result from sideropenic anemia with reticuloendothelial siderosis. At recurrence in clinical course of uterine cancer, no rise of serum ferritin was observed. From our results, serum ferritin does not seem useful for the diagnosis and following up of uterine cancer.

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We measured the red cell enzyme activities in 556 cases with hematologic disorders, most with hemolytic anemia, during the past 9 years. About half of them were from patients for whom a hematologic diagnosis had already been considered to be established or could readily be established from the data submitted with the sample. Of the 254 samples which were previously undiagnosed cases with congenital nonspherocytic hemolytic anemia, a diagnosis of erythroenzymopathy was established in 88 cases, and 10 cases were diagnosed as unstable hemoglobin. Many other patients (156 cases) had no red cell enzyme defect or unstable hemoglobin.
Although some of the samples in which we were unable to detect an enzyme deficiency or unstable hemoglobin seem to be due to our erroneous diagnosis, there may be the existence of the unknown metabolic pathway which is important to maintain the red cell function.
Pyruvate kinase deficiency and G6PD deficiency were the commonest erythroenzy-mopathies in Japan. The incidence of pyrimidine 5'-nucleotidase deficiency was thought to be high.

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A clinical trial was made for Simplate II comparing with the Template and the Duke methods.
The mean bleeding times were 4.36 minutes (S. D.=1.33, n=30, healthy volunteers) with Simplate II, 6.35 minutes (S. D.=1.99, n=14) with the Template method, and 2.92 minutes (S. D.=0.90, n=14) with the Duke method. Those with both Simplate II and the Template methods revealed a good correlation (R=0.665, P<0.01), however, those with Simplate II and the Duke method showed a poor correlation (R=0.479, P<0.05).
Bleeding times before and after aspirin ingestion (dosis ranging from 7 mg/kg/day to 50 mg/kg/day, 3 times daily, for 3 days) were measured by these 3 methods in 4 adults and found that those after aspirin ingestion, irrespective of low or high doses, were significantly prolonged in both Simplate II and the Template method, but not so much prolonged with the Duke method.
In 6 patients with platelet dysfunction (Bernard-Soulier syndrome, thrombasthenia, von Willebrand disease), prominent prolongation of bleeding times was observed in both Simplate II and the Duke method.
Simplate II is a sufficiently reliable device of bleeding time, and simple to use. However, it sometimes leaves a scar on the forearm. Therefore, we should carefully consider the clinical application of this device.

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Sensitive enzyme immunoassay system for measurement of β-thromboglobulin (β-TG) in plasma was prepared with purified antibody to β-TG and β-D-galactosidase from Escherichia coli as labeled substance. The antibodies were purified with an immunoaffinity chromatography form the antiserum, which had been produced in rabbits by injecting purified β-TG. The assay system consisted of the polystyrene bead with immobilized antibody F (ab')₂ fragments and the antibody Fab' fragments labeled with β-D-galactosidase. The assay was highly sensitive with β-TG measurable at levels as low as 10 pg/assay tube. β-TG levels in plasma was measured with 2.5 μl samples. Under the conditions, coefficient of variation for within and between run was less than 8% and the results obtained with the enzyme immunoassay correlated well with those with a radioimmunoassay. The plasma β-TG levels in healthy subjects determined with the present assay system were 38.4±6.2 ng/ml and the levels were markedly high in patients with myeloproliferative syndrome and in patients with DIC.

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Recently, it was shown that the synthetic polymers, poly rA·oligo dT and poly rCm·oligo dG were effective template-primers for assaying reverse transcriptase originated from RNA tumor virus. These template-primers have been used for distinguishing reverse transcriptase from DNA dependent DNA polymerase (DNA polymerase β or γ). Reverse transcriptase activities were estimated in leukemia and hemopoietic dysplasia. They were five cases of AML, one case of APL, one case of AMoL, two cases of CML, three cases of hemopoietic dysplasia, three cases of ALL and one case of hairy cell leukemia. DNA polymerase activities and the ratio of DNA polymerase activities were measured by different template-primers, poly rA·oligo dT, poly rCm·oligo dG and poly dA·oligo dT.
As a result, reverse transcriptase activities were found in one case of AML, two cases of hemopoietic dysplasia, one case of APL and one case of hairy cell leukemia. It is especially noted that reverse transcriptase activities have been found in two cases of hemopoietic dysplasia. They should be measured repeatedly during the clinical course of these patients. It is inferred from these results that RNA virus might be concerned to the etiology of some cases of human leukemia.

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In this study previous knowledge of PAS reactivity of acute leukemia is extended to the ultrastructural level by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) techinique showing the intracellular distribution of the reactive components and the existence of reactive sites not evident with the light microscopic level.
PA-TCH-SP method, which visualized complex carbohydrates with hexose containing vicinal glycol group like PAS staining, stained the granules, glycogen, Golgi apparatus, and cell coat of peroxidase-negative acute leukemia cells. Granules were classified 4 types by PA-TCH-SP reactivity such as clustered dense body-like granules, scattered dense body-like granules, large granules and unevenly-stained abnormal granules. Generally, the PA-TCH-SP reactivity of the clustered dense body-like granules was stronger than that of the scattered dense body-like granules. Unevenly stained granules were observed in poor prognostic group of the leukemia. Glycogen was distributed evenly in the cytoplasm with small particles, or clustered to be a large pool. Coarse granular PAS staining was clustered glycogen or clustered dense body-like granules by PA-TCH-SP on ultrastructural level, and diffuse PAS positivity was abundant glycogen particles evenly distributed in cytoplasm. Small amount of glycogen particles and the scattered dense body-like granules were PAS negative on light microscopic level because of weak resolution.
The PA-TCH-SP method of electron microscope is valuable technique for cytochemistry of complex carbohydrates in peroxidase-negative acute leukemia.

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A 54-year-old male was admitted to the Tokai University Hospital because of intramuscular and subcutaneous hemorrhage on July 15, 1980.
Partial gastrectomy had been performed on him because of gastric ulcer in June, 1977 and stomal ulcer was re-operated in December, 1979, when he received blood transfusion, sulbenicillin and penicillin G.
Physical examination revealed ecchymoses over the right arm and the left hand. Coagulation studies showed prolonged partial thromboplastin time and decreased factor VIII activity (1.8%). Inhibitor to factor VIII was demonstrated by 48 Bethesda units. Gel filtration of the patient plasma on Sephadex G-200 revealed the inhibitor activity was present in the second protein peak.
He was initially treated with prednisolone and cyclophosphamide without any improvement of factor VIII activity. Then a large dose of factor VIII (5,000 U) and a single infusion of cyclophosphamide (500 mg) were given, which resulted in disappearance of the inhibitor and improvement of the coagulation profiles, although he developed acute hepatitis and the concentration of factor VIII inhibitor increased again 6 weeks later.

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Patients with hypoplastic leukemia in advanced age are recently increasing in number, but we don't have any established design for the treatment.
A 64-year-old female of hypoplastic leukemia successfully treated with continuous intravenous drip infusion of Cyclocytidine (450 mg/day) and prednisolone (30 mg/day) for ten days and induced into complete remission is reported.
Although she suffered from high fever due to infection and gastrointestinal bleeding during the treatment, these were overcome by intensive supportive therapy. After an induction of complete remission, bone marrow still remained hypoplastic.
As a side effect, the patient had a slight parotic pain which soon subsided.
Cyclocytidine has the following characteristics; high therapeutic index, low toxicity (e. g. low myelosuppression), and schedule dependency. On account of these features, this drug is appropriate very well in cases of aged and hypoplastic leukemia like in this case.
As for the dosage and duration of the treatment with this drug further study may be necessary. In addition to chemotherapy intensive and appropriate supportive therapy is necessary and immunopotentiator should be administered.

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A 31-year-old female was referred to our hospital on March 19, 1980, because of purpura and hypermenorrhea over one year with no response to prednisolone therapy. Her past history was noncontributory. Physical findings on admission; scattered petechiae were observed on the upper and lower extremities and the neck. She had a diffuse goiter and an exophthalmus, but no hepatosplenomegaly. Laboratory examination revealed a marked thrombocytopenia (10,000/mm³) with normal red cell and leukocyte counts in peripheral blood. Bone marrow aspiration was unremarkable with normal megakaryocyte count (50/mm³). Serum levels of T₄ (13.4μg/dl) and T₃ (2.17ng/ml) were both elevated and that of TSH (less than 0.3μIU/ml) depressed. TRH test was unresponsive. Serologically, anti-platelet antibody was negative, thyroid test (×80²) and microsome test (×160²) were both positive.
On the diagnosis of ITP associated with Basedow's disease, combination therapy of prednisolone and methimazole was instituted with a rapid increase of platelet count to normal. When she became euthyroid, prednisolone therapy was tapered off. The platelet count was sustained normal.
The mechanism of the co-occurrence of ITP and Basedow's disease was discussed.

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A case of nonspherocytic hemolytic anemia associated with abnormalities in membrane lipids was described. The 19-year-old boy was consulted because of jaundice and left hypochondralgia. Splenectomy had been performed two years before. On admission, hemolytic anemia was evident, having 12.3g/dl of hemoglobin, 208‰ of reticulocytes, indirect bilirubin of 5.5mg/dl and the presence of target red cells in peripheral blood smears. The patient's red cells had an absolute increase in erythrocyte phosphatidylcholine. There was no evidence for abnormalities in glycolytic enzymes, hemoglobin analysis, liver function test or plasma lipids. Osmotic fragility was decreased, Coombs' test was negative and ⁵¹Cr labeled red cell survival was 6.2 days. It was concluded that the hemolytic anemia was due to an intrinsic red cell defect, which was elevated phosphatidylcholine in red cell membrane lipids. There are only two reports of high phosphatidylcholine hemolytic anemia in the world.

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The patient is a 59-year-old female who was operated for right side pontocerebellar angle tumor on May 22, 1977. The pathological diagnosis of the tumor was acoustic neurinoma. Blood examination revealed leukocytosis in April 1979, and the patient was diagnosed as having chronic myelogenous leukemia by bone marrow films and a chromosome analysis. The patient was admitted to Kasumigaura Hospital, Tokyo Medical College because of fatigability on January 12, 1981, and was diagnosed as having primary aldosteronism together with chronic myelogenous leukemia by endocrinological and hematological examinations. Cytogenetic analyses of cells from the bone marrow showed a Ph¹ translocation, i. e., a translocation between chromosomes No. 9 and No. 22 and a pericentric inversion of the heterochromatic region of chromosome No. 1. The chromosome analysis of PHA-stimulated lymphocytes revealed the pericentric inversion of the chromosome No. 1 to be a constitutional chromosome abnormality.
Heteromorphism of chromosome No. 1 and its possible association with malignancy is discussed.

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Two cases of pyruvate kinase variants were reported. These patients were a girl, aged 19 (case 1), and a boy aged 16 (case 2). The parents of both cases are first cousins.
Case 1: The jaundice was noted shortly after birth. She was admitted to our hospital and diagnosed nonspherocytic hemolytic anemia at 12 years old. The liver and spleen were palpable 1.5cm below the respective costal margins.
Case 2: He had been pale since birth. Jaundice and hepatosplenomegaly were noticed at 11 years old. At the age of 12, he had a splenectomy for the tentative diagnosis of Banti syndrome.
In both cases, red cell PK was lowered. Red cell glycolytic intermediates showed high PEP, 2-PG, 3-PG, 2, 3-DPG and low ATP levels.
These variants were designated PK Aizu (case 1) and PK Fukushima (case 2).
Enzymatic characteristics of these cases have been reported elsewhere (Brit. J. Haemat., 45: 575, 1980).

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A 64-year-old male was admitted with a chief complaint of giant hematoma on left lateral site of the chest on May 12, 1980. On admission, leukocytosis, slight hypochromic anemia, marked thrombocytosis and hepatosplenomegaly were noted. Serum LDH, uric acid and potassium levels were high.
Chromosome analysis was normal and NAP score was slightly elevated. He had normal marrow cellularity with erythroid hypoplasia and normal megakaryocyte count. He was diagnosed as primary thrombocythemia. He showed remarkable hemorrhagic diathesis with hematoma in increasing size and new hematoma formation. Hemonetics blood processor model 30 was applied to the patient for the safe and rapid mechanical removal of platelets. Platelets of 4-5×10¹² were removed by this procedure per a time. Aggregability and release ability were studied on the platelets obtained from patient before and after thrombocytopheresis. Before thrombocytopheresis, ADP-, collagen-, epinephrine- and ristocetin-induced platelet aggregation were markedly impaired, and these functions did not recover by PRP-dilution with patient plasma to normal platelet count. Function tests of platelets obtained from patient after thrombocytopheresis recovered to fairly normal values.
These results indicated the possibilities that something in vivo other than platelets might affect the platelet functions. Thrombocyte count was fallen to about 200×10⁴/cmm and serum LDH, uric acid and potassium were decreased to normal level by serial thrombocytopheresis. Then carboquone was administered and thrombocyte count became to normal level.
Thrombocytopheresis is a useful method for treatment of hemorrhagic diathesis and for prevention of uricemic nephropathy, which might develope after chemotherapy, in primary thrombocythemia.

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