Immunological studies were performed on patients with aortitis syndrome, Buerger's disease and Raynaud's syndrome. Immunological analysis of serum protein, delayed hypersensitibity, T-cell, stimulation index by phytohemagglutinin and leucocyte migration inhibition tests were investigated.
Definite increase of α₁-antitrypsin and haptoglobin, C3 and C4 was found in patients with aortitis syndrome, especially in active stage, while not in patients with Buerger's disease except those with ischemic ulcer.
High incidence of negative skin tests, and significant decrease of T-cell and stimulation index were found in aortitis syndrome, while not significant in Buerger's disease.
It was concluded that cell-mediated immunity was significantly decreased in the former.
These findings are suggestive of the participation of immunological mechanism in development of aortitis syndrome, although not denied in Buerger's disease.
Brief review of recent advances in this field was also given.

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The role of factor XIII in fibroblast growth was investigated by means of 3T6 cell culture on fibrin plates prepared from purified fibrinogen and factor XIII concentrate in the presence of various concentrations of the factor XIII inhibitor, glycine ethyl ester (GEE). Electrophoretic analysis of the reduced fibrin preparations indicated that γ-chains were almost exhaustively dimerized in 2 hours regardless of the inhibitor and α-chain crosslinking ceased in about 6 hours. 1-5 mM GEE failed to inhibit γ-γcrosslinking but did inhibit α-polimerization depending on the concentration. 3×10⁴ living cells were seeded in Dulbecco-Vogt-Modified Eagles medium supplemented with 10% heat-inactivated fetal calf serum on a fibrin plate 3 hours after its clotting in the culture bottle and the cell growth rate was determined from the living cells count after cultivation for 48 hours.
The cells were poorly proliferated on fibrin prepared in the presence of the inhibitor, but grew well on fibrin without the inhibitor and even with the inhibitors added at 3 hours after fibrin clotting. The cell growth rate was proportional to the decrease in α-chain content, i. e. the densitometrically determined α/β for the electrophoretic pattern. Thus 3T6 fibroblast proliferation appeared to depend essentially on the extent of α-chain crosslinking of its supporting fibrin matrix after the completing of γ-γ crosslinking.

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Recently, it has been reported that metabolites of arachidonic acid in platelets play an important role in the first step of thrombus formation and that prost-acyclin, one of metabolites of arachidonic acid in vascular walls inhibits platelet-aggregation. However, it is still unknown how the above metabolites of arachidonic acid are involved in platelet-aggregation.
In this paper, inhibitory mechanisms of platelet-aggregation by vascular walls were studied by using bovine carotid arteries. It has been reported that prostacyclin produced by vascular walls could elevate the intracellular cyclic AMP and that some agents which elevate the cyclic AMP have an inhibitory effect on platelet-aggregation. On the other hand, some substances which are Ca⁺⁺-chelators, such as EGTA inhibit the platelet-aggregation. Then, the effect on platelet-aggregation of Verapamil which was thought to inhibit Ca⁺⁺-influx was studied. And, it was investigated if the elevation of intracellular cyclic AMP might have any effects on a behavior of the intracellular Ca⁺⁺, by using subcellular fractions from human platelets.
The results obtained in this paper were as follows: 1) Microsomes from bovine carotid arteries inhibited collageninduced platelet-aggregation by producing PGI₂. 2) Verapamil inhibited the collagen-induced platelet-aggregation. 3) Platelets incubated with both arterial microsomes and Verapamil, each of which alone was not enough to inhibit the collagen-induced platelet-aggregation, could not be aggregated. 4) Cyclic AMP level in platelets incubated with both the arterial microsomes and Verapamil was higher than that incubated with either of them. 5) The cyclic AMP-stimulated calcium uptake by subcellular membranous components was closely correlated with their cyclic AMP-stimulated endogenous phosphorylation.
In summary, it is suggested that the intracellular calcium ion of platelets with anti-aggregating agents may be too low to activate the calcium sensitive contractile protein for the thrombus formation.

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The effect of prostacyclin (PGI₂) on platelet functions was investigated using synthesized PGI₂ sodium, which was a gift of Dr. Schölkens in Hoechst Research Institute in West Germany. Platelet aggregation induced by ADP (4μM), epinephrine (2μg/ml) and arachidonic acid (2mM) was markedly inhibited by PGI₂ at a concentration of 10nM. Minimal concentration of PGI₂ to inhibit platelet aggregation induced by collagen (2μg/ml) or A23187 (10μM) was 2.5nM and 20nM respectively. The inhibitory activity of PGI₂ was found to be five times potent than that of PGE₁. PGI₂ was less potent in medium than in plasma and its potency appeared to be dependent of plasma concentration. Plasma factor to stabilize PGI₂ activity was not determined, but not albumin, fibrinogen nor γ-globulins. Minimal concentration of PGI₂ to inhibit A23187-induced platelet aggregation was 20nM, while that of TMB-8, intracellular calcium antagonist, was 1mM. However, the addition of 0.1mM TMB-8 and 5nM PGI₂ markedly inhibited platelet aggregation induced by A23187, demonstrating the synergistic effect of PGI₂ and TMB-8 on platelet aggregation. ¹⁴C-serotonin release induced by thrombin (1u/ml) in tris-buffered saline containing 2mM EDTA, was inhibited by 20-50nM PGI₂, while that by A23187 (1μM) was not inhibited even by 500nM PGI₂. These results gave additional informations to characterize the inhibitory action of PGI₂.

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TEI-2117 (I), 2-(3-benzoyl-2, 5-dimethylpyrrole) propanol, is a novel compound synthesized by our laboratories. The effect of I was studied on human platelet aggregation and on prostaglandin biosynthesis in vitro comparing with indomethacin (IM). The compound (I) was more potent inhibitor than IM in arachidonic acid-induced platelet aggregation. I had similar inhibitory activity to that of IM in aggregation by collagen. It also inhibited the secondary phase of aggregation induced by epinephrine. Inhibition of the release of rabbit aorta contracting substances was also observed by preincubation of PRP with I. However, I was less active in inhibition of prostaglandin E₂ formation when bovine seminale vesicles were used as an enzyme source. In the study of prostaglandin biosynthesis using human platelet microsome and ¹⁴C-arachidonic acid, the regulatory pattern of prostaglandin biosynthesis by I was different from that by IM. The effect of I on platelet aggregation was further studied in extra vivo. When I was administered subcutaneously (10mg/kg) or orally (100mg/kg) to guinea pig, 98 percent and 70 percent inhibition of platelet aggregation induced by arachidonic acid, was observed, whereas aspirin was nearly equipotent to I at the same dose. These result suggest that I may play in vivo effect on prevention of thrombus formation through antagonizing arachidonic acid metabolism in platelets.

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ADP-induced platelet aggregation at final concentration of 2μM (ADP-Agg.) was performed according to the method by Born using a Bryston aggregometer model 350. The sensitivity of the platelets was expressed in terms of the maximum percentage of light transmittance to that of platelet-poor-plasma within 5 minutes after the addition of ADP. Peripheral neuropathy was evaluated by motor nerve conduction velocity (M. C. V.). Retinopathy was evaluated by usual ophthalmoscopy.
The subjects studied were 20 age-matched healthy subjects and 47 diabetic patients. These 47 diabetic patients were divided into following four groups according to their M. C. V. and with or without retinopathy. N (-) indicates the patient whose M. C. V. was not less than the mean M. C. V. of age-matched normal subjects. R (-) indicates the patient whose fundoscopic findings revealed nothing remarkable as to diabetic retinopathy, e. g. microaneurysm, retinal hemorrhage etc. The results of two-sample t test of ADP-Agg. were significant between the groups listed as follows. The figures in parenthesis indicate the number of subjects.
P<0.001…Control (20)<Diabetics (47)
P<0.01…R(-) N(-) (16)<R(+) N(+) (13)
R(+) N(+), R(-) N(+), R(+) N(-) (31)>R(-) N(-) (16)
P<0.05…R(-) (22)<R(+) (25)
N(-) (28)<N(+) (19)
R(+) N(-) (12)<R(+) N(+) (13)
R(-) N(-) (16)<R(-) N(+) (6)
These results indicate that platelets of N (+) patients are more sensitive to ADP than those of N (-) patients regardless of retinopathy.
In the diabetic patients FBS and ∑BS (50gOGTT up to 2hrs.) correlated with ADP-Agg. but the serum cholesterol, triglycerides β-lipoprotein and ΔIRI₃₀/ΔBS₃₀ did not.
From the foregoing, it appears that in the diabetics hyperglycemia per ce might play an important role in the observed platelets abnormalities of patients with peripheral neuropathy.

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The platelet production increases in stroke-prone SHR (SHRSP) after 10 weeks of age, probably due to increased platelet consumption compared with those in stroke-resistant SHR (SHRSR) (Koganemaru, S., et al. Jap. Heart J. 18: 598-599, 1977). We extended our study to see the effect of aging, blood pressure, platelet-inhibiting drug and Vitamin E (Vit. E) on this kinetic change. SHRSP (A₁-sbF_37-39) and SHRSR (B₁F_36-38) with matched age (5 weeks, 10 weeks and 9-12 months) and sex were used. A dose of 2μCi/100Gm; ⁷⁵Se-methionine (⁷⁵SeM) was injected via tail vein. The platelet survival time was normal in both groups at 6 weeks of age, but at 9-10 weeks, it was shorter (3.7 days) in SHRSP. This tendency was more marked in SHRSP at 9-12 months with corresponding increase of maximum uptake into platelets. The maximum uptake was not affected in both salt loading group and clonidine group in SHRSP. A significant decrease was observed in dipyridamole group (0.096±0.020, p<0.05). The platelet survival time was shortened in Vit. E deficient group of SHRSR; 3.0 days at 9-10 weeks. In Vit. E sufficient group of SHRSP, the platelet survival time was corrected to normal. The maximum uptake of ⁷⁵SeM was increased in both Vit. E deficient group of SHRSP (0.119±0.010 vs 0.192±0.030, p<0.01) and SHRSR (0.071±0.016 vs 0.119±0.018, p<0.01). Thus the platelet production is increased and survival time is shortened in SHRSP with aging after 10 weeks of age. The increased platelet production is not affected by blood pressure but it can be suppressed by treating with dipyridamole or giving Vit. E before 10 weeks of age. Therefore it is strongly suggested that there is already a vascular alteration in SHRSP at 10 weeks of age, when blood pressure begins to elevate.

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Changes in platelet function under an anesthesia during surgical operation are not yet clarified. In this paper, the effect of anesthesia with nitrous oxide on platelet functions was examined, and the following results were obtained: 1. ADP induced platelet aggregation did not show any significant change before and after a premeditation with opistan in 20 patients. 2. Blood was collected from 15 patients before and 30 and 90min after a begining of anesthesia under an administration of nitrous oxide for gynecological operation and postoperatively. Platelet count did not show any significant change. Bleeding time was prolonged slightly at 90min after the beginning of anesthesia, with a statistical significance. But no case showed a change beyond the normal range of bleeding time. ADP induced platelet aggregation and platelet factor 3 activity showed no significant change during the observation period. 3. A direct effect of nitrous oxide on platelets was examined in vitro. Nitrous oxide was pumped through the 18G teflon tube into a 10ml siliconized glass tube containing 2ml of the PRP or 3ml of washed platelets for 5 minutes or 30 minutes. The platelet aggregations induced by ADP or collagen were compared with the control, but there were no significant difference in the rates of aggregations. We measured the levels of malondialdehyde (MDA) and c-AMP in washed platelets after incubation for 5 minutes and the level of MDA did not change.
In conclusion, this study revealed that there is no significant inhibition on platelet function under anesthesia with nitrous oxide and operation.

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An assay system was developed to measure anti-platelet IgG in serum by adapting the complement lysis inhibition test reported by Dixon et al., which measured the IgG amount bound to platelets of ITP patients. O type platelet suspensions (200, 000/cmm) washed with PBS were obtained from 5 healthy volunteers. Fixation with 2% formalin and washing with PBS were followed by adjustment of the platelet concentration to 100, 000/cmm. Using a vacuum pump, the formalin fixed platelets were dried in porystyrene tubes (Shionogi tube, 10×70mm) which were activated with poly-L-lysine beforehand; the tightly covered tubes were kept at 4C until use.
Even after several washings, SEM showed a homogenous monolayer of dried platelets with a few pseudopods attached to the bottom. Using standard IgG solution, the IgG amount in the serum was calculated by measuring the absorbance of supernatant at 541nm.
A 77-year-old man with acute granulocytic leukemia had 3.8μg IgG per ml of his serum. IgG in one half and one fourth volumes of the serum were assayed as 2.0 and 0.8μg/ml, respectively, indicating good reliability.
Five healthy men showed less than 2μg IgG/ml, whereas 6 of 11 aplastic anemia patients between 3 and 6μg/ml. IgG in 4 of 9 ITP cases and in 5 of 9 leukemia cases were between 3 and 4μg/ml. The IgG amount measured successively in a case of aplastic anemia after transfusions of 2 units of leukocyte poor red cells and 5 units of platelet concentrates every week changed from a lower limit of the assay to 5.8μg/ml.
This method can be used as a screening test for anti-platelet IgG antibody in serum.

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The plasma concentration of the platelet-specific protein β-thromboglobulin (β-TG) was measured by the use of, “β-TG RIA kit” (The Radiochemical Centre, Kakenkagaku) in patients with various disorders, including 12 patients with thromboembolism, 9 with prosthetic cardiac valves, 10 with diabetes mellitus, 18 with various thrombocytopenias, 15 with various thrombocytosis and one of each with von Willebrand's disease and vascular purpura as well as in 21 healthy control subjects. The mean concentration in control subjects was 23ng/ml (range 8-50, S. D. 14). The inter-assay coefficient of variation (CV) and the intraassay CV of a single plasma sample were 17.3% (n=7) and 12.1% (n=12), respectively. Except for disseminated intravascular coagulation (DIC), all thrombocytopenic patients showed normal or low β-TG levels. It was to be noted that β-TG was not increased in idiopathic thrombocytopenic purpura (ITP) in which immunologically damaged platelets were presumed to be removed from the circulation in such a way that plasma β-TG was not elevated. Plasma β-TG was increased in patients with primary thrombocytosis (myeloproliferative disorders), and similar but less marked change was observed in secondary thrombocytosis. Raised β-TG levels were detected in most patients with recent thromboembolic episodes but not in those with old ones. One prosthetic valve patient and 3 diabetic patients with recent vascular complications showed high β-TG levels but those without such complication had normal values. A patient with von Willebrand's disease showed a normal β-TG level and that with vascular purpura had the raised ones. The studies documented a progressive rise and fall in plasma β-TG concentration following the onset of acute thromboembolism or DIC. However, persistently elevated levels were detected in patients with essential thrombocythemia and vascular purpura without clinical manifestations of thromboembolism. The investigations suggest that radioimmunoassay of β-TG is a useful test to aid the diagnosis and treatment of patients with thromboembolic disorders as well as to understand pathophysiology of some platelet and vascular disorders.

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In myeloproliferative disorders, bleeding tendency and thrombosis were encountered occasionally, in the presence of extremely high platelet counts. Platelet membrane glycoproteins as receptor sites were studied by SDS disc electrophoresis and two dimensional electrophoresis in chronic myelogenous leukemia (CML, 7 cases), polycythemia vera (PV, 2 cases), primary thrombocythemia (PT, 2 cases) and myelofibrosis (MMM, 1 case). In CML group any band of abnormal molecular size was not found but the percent of GP I to all PAS stained peaks was reducedd and GP III was increased statistically. Four peaks labelled with ¹²⁵I-Na by lactoperoxidase were appeared in accordance with PAS staining in these disorders.
In GP I peak, the ¹²⁵I incorporation rate was not different between normal and myeloproliferative disorders. The percent of PAS staining intensity was decresed in CML but increased in PT, PV and MMM. In GP III peak of CML, the increased percent of PAS staining and the decreased ¹²⁵I incorporation rate were proved compared to the control. In MMM, PV and PT, it was shown that intensity percent of PAS staining was normal but ¹²⁵I incorporation rate was decreased against GP III.
To analyse the dissociation between PAS stain and ¹²⁵I incorporation rate, the two dimensional electrophoresis was conduced with non reduced-reduced system. The autoradiographic pattern of this electrophoresis showed seven ¹²⁵I incorporated spots (GP Ia, Ib, Ic, IIa, IIb, III and IV) in normal. The GP Ib was showed as an unclear spot and GP IV was divided in two spots in 4 of 6 CML cases. The GP IV appeared as a faint spot in two PT cases. In MMM and PV, the autoradiographic patterns were similar to those of CML cases.
Association of these findings in surface glycoproteins with abnormalities of epinephrine and collagen induced aggregation in these disorders are under investigation.

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Activation of coagulation and inhibition of fibrinolytic system during pregnancy have been well documented. Also chronic DIC is often accompanied with preeclampsia.
In our series of DIC study, coagulation-fibrinolysis system, platelet count and platelet function were measured in non-pregnant, normal pregnant females and patients with preeclampsia. The results obtained were as follows:
1) Coagulation factors, except for factor XIII, were increased to 130-300% of normal and fibrinolytic activity was decreased during pregnancy.
2) There were no differences in fibrinogen, plasminogen, platelet counts and platelet aggregation between normal pregnants and patients with mild preeclampsia. In severe preeclampsia, however, fibrinogen and plasminogen levels were low as compared with those of normal pregnants or mild preeclampsia.
3) The platelet counts and maximum aggregation rate by ADP were decreased significantly in severe preeclampsia. Gestosis Index and platelet aggregation rate showed a negative correlation (r=-0.782).
4) The mean β-thromboglobulin level showed a slight rise during pregnancy. It was increased in severe preeclampsia.
It was postulated that the decrease of maximum aggregation rate in severe preeclampsia was produced by increased number of platelets of which function was depressed as a result of the enhanced release reaction.

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The authors have reported that when the patients with ischemic heart disease (IHD) were loaded with isometric exercise, platelet sensitivity to ADP-aggregation and plasma von Willebrand factor (vWF) level increased. In this paper, influence of isometric exercise on plasma β-thromboglobulin (β-TG) level was investigated.
MATERIALS AND METHODS: Eleven patients with IHD (Table 1), none of them were in acute stage, and their age-and sex-matched healthy controls were loaded with handgrip isometric exercise of 50% maximal contraction using a Smedley's handgrip dynamometer for 2 minutes. Plasma β-thromboglobulin level, platelet sensitivity to ADP-aggregation and plasma vWF level were measured before and immediately after exercise and the values were compared statistically with a paired comparison procedure. Plasma β-TG was measured using β-TG Ria kit (The Radiochemical Centre, England). Platelet sensitivity was measured using a original method to detect microscopically the highest dilution (n) of serially two-fold diluted ADP solution (2^-nmg/ml) to give platelet aggregation. When the endpoint was 2^-10mg/ml, for example, the sensitivity was expressed as 10. Plasma vWF was also tested using a original device. The maximal dilution (n) of the serially two-fold diluted test plasma (1:2ⁿ) to give ristocetin-induced platelet aggregation was detected microscopically and n was defined as plasma vWF titer.
RESULTS (Fig. 1): 1) IHD: In 8 patients plasma β-TG level increased after exercise and the change was significant (p<0.05) (before exercise 58.0±17.71, after exercise 70.4±21.21). Platelets were aggregated even by 2 to 4 times diluted ADP after exercise and platelet sensitivity increased significantly (P<0.05) (bef. 9.8±0.32, aft. 10.4±0.34). Plasma vWF titer also increased significantly (p<0.05) after exercise (bef. 5.0±0.19, aft. 5.6±0.19). 2) Controls: No significant changes.
COMMENT: The results suggest that isometric exercise may induce in vivo activation and release reaction of platelets in IHD patients. It might be attributable to a forced passage of platelets through stenotic portion of atherosclerotic coronary vessel during isometric exercise.

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In order to detect the activated platelets, plasma beta-thromboglobulin (β-TG) levels and nucleotide pools in platelets were determined in blood samples from 34 diabetic patients with various vascular complications i. e. retinopathy, nephropathy, ischemic heart disease, cerebral thrombosis and obstruction of arteries in the extremities.
β-TG levels were measured using RIA kit (Radiochemical Center, Amersham). Higher plasma, β-TG levels than 50ng/ml were found in seven patients (21%).
Nucleotide pools in platelets were determined with high pressure liquid chromatography. ATP and ADP levels in platelets were decreased in 47% and 35 of the patients, respectively. ATP/ADP ratio in those cases was ranged between 0.9 and 2.8. Amount of GTP in platelets was elevated in 3 cases and decreasedd in 2 cases. All three cases whose plasma β-TG levels elevated more than 100ng/ml showed decrease of both ATP and ADP in the platelets.
Measurement of nucleotide pools in platelets is a more sensitive method to detect the evidence of platelet activation in patients with arteriosclerosis.

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Concentrations of β-thromboglobulin in plasma (β-TG) were measured in 69 healthy subjects over age sixty and in 157 patients with various diseases.
Mean value (and standard deviations) of β-TG in the healthy elderly were 32±20ng/ml. β-TG levels in these subjects increased slightly with age.
In 5 patients with acute or recent myocardial infarction, in 7 cases of acute or recent cerebral infarction, or in 5 cases of transient ischemic attacks, moderate elevations of β-TG concentrations were found, although β-TG levels increased only sightly in 42 cases of old cerebral infarction. In 63 diabetics, an increase in β-TG was observed. Concentration of this protein were higher in patients with retinopathy.
In 8 cases of DIC, β-TG increased moderately, although in a case of megaloblastic anemia, in whom DIC developed despite of impaired production of platelets in bone marrow, concentration of β-TG remained within normal limits. In 5 patients with deep vein thrombosis and in 3 cases of nephrotic syndrome, β-TG increased markedly. In 3 cases of cancer without evidences of DIC, β-TG increased slightly. In 5 cases of aplastic anemia and 2 cases of acute leukemia, in whom platelet production was remarkably suppressed, markedly decreased levels of β-TG were observed, whereas levels of this protein were within normal limits in a case of ITP and in a case of liver cirrhosis with thrombocytopenia. Conversely, a stikingly elevated level of β-TG was observed in a case of essential thrombocythemia.
These findings suggest that an increase of β-TG in plasma indicates thrombotic tendency, although possibilities that short half-life of this protein may cause subnormal values in cases of thrombosis or even in patients with DIC and that bad venipuncture causes false elevation of β-TG cannot be ruled out.

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When plasminogen was mixed with urokinase (UK), monodextran sulfate (MDS) and S-2251, the hydrolysis of S-2251 was not influenced in the presence of MDS. However, MDS increased the activation of plasminogen in the plasma, when MDS was added to the mixture of plasma, urokinase and S-2251. Although the clot lysis time induced by UK-activated plasmin was shortened with the increse in the concentration of MDS, the amount of plasmin which was measured by the hydrolysis of S-2251 was decreased. On the other hand tranexamic acid increased the activation of plasminogen by UK, which was measured by the hydrolysis of S-2251, but no clot lysis was observed. The presence of 10μM of tranexamic acid resulted in decrease in the activation rate of plasminogen in the clot, however the addition of MDS prevented this decrease. Thus tranexamic acid and MDS had opposite effects on fibrinolysis and hydrolysis of S-2251 in the plasma and clot.

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We had already demonstrated that the medical materials immobilized urokinase possessed the effective fibrinolytic activity in vitro and in vivo. Now, we planed the clinical application of these materials.
This report covers the clinical trials.
These materials were immobilized urokinase through the grafted copolymer such as polyethyleneimine and gantrez by peptide linking and disinfected by ethylenoxide gas or gamma rays.
These results were as follows:
I. The fibrinolytic activity of nylon immobilized urokinase was moderately damaged by the contact with disinfectants or with body tissues. The remained fibrinolytic activity reached 40-70% after the treatment of various disinfections and 57-85% after the placement in abdominal cavity of rat.
II. The clinical trials were performed in three groups.
1) The nylon suture material which sawed the vessel wall was prepared by urokinase and applied in 12 operations of ten patients who underwent the thromboendarterectomy in small artery. Two patients failed in reconstruction after a few months.
2) The dacron thread or nylon thread which was implanted in subcutaneous tissue was prepared by urokinase and applied in seven operations of six patients who underwent the Handley's method for drainage in lymphedema. Some patients were in good clinical courses. In two patients, some pieces of dacron thread were removed by the reasons they were contaminated or reimplanted. Their microscopic findings were shown in Figure 1 (A), (B) and (C).
3) The nylon tube or perplene tube which was inserted into caval vein was prepared by urokinase and applied in 12 infusions of 12 patients who underwent the intravascular hyperalimentation. The material had the favorable thromboresistant quality. Their macroscopic findings were shown in Figure 2.

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Fibrinopeptide A (FPA) was measured in human plasma by a radioimmunoassay kit (IMCO). In 14 normal individuals this assay revealed a mean FPA level of 1.5ng/ml. 121 patients were divided into 6 groups by the levels of fibrinogen and FDP as follows.
A. Patients with elevated fibrinogen levels and elevated FDP levels (n=16)
B. Patients with elevated fibrinogen levels and normal FDP levels (n=13)
C. Patients with normal fibrinogen levels and elevated FDP levels (n=17)
D. Patients with normal fibrinogen levels and normal FDP levels (n=44)
E. Patients with reduced fibrinogen levels and elevated FDP levels (n=7)
F. Patients with reduced fibrinogen levels and normal FDP levels (n=24)
The mean FPA levels were, 9.4ng/ml in group A, 3.7ng/ml in group B, 3.9ng/ml in group C, 5.3ng/ml in group D, 4.1ng/ml in group E, and 4.8ng/ml in group F, respectively. These results indicated that elevated FPA levels were found in the patients with normal fibrinogen levels and normal FDP levels. Considering FPA levels from clinical disorders, elevated FPA levels were found in patients suffering from Dis-seminated intravascular coagulation, leukemia, malignant neoplasma, pulmonary embolism and other thrombotic disorders. After injection of heparin in the patients with these disorders, FPA level was reduced rapidly into the normal range.
These results suggest that FPA is most sensitive indicator of thrombin action, and provides direct information concerning the effectiveness of therapy in preventing fibrin formation.

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The purpose of this study is to elucidate whether LMWD and combined use with UK have the prophylactic effect of thrombus formation and thrombolytic enhacement or not. This study was carried out by Chandler's loop method.
One hundred thirty ml of blood sample were respectively taken from two healthy young volunteers at the rate of one volume of 3.8% citrate and nine of blood. One ml of citrated blood was poured into each Chandler's loop and then 0.1ml of 0.25M calcium chloride was added. An artificial clot was formed after 30 minutes rotation of a loop equipped to a cylinder which rotates 12 times per minute, holding 80 degree of angle upwards from the horizontal surface. After a clot formation, various concentration of LMWD and UK was added into each Chandler's loop. Each clot was weighed after four hours rotation. Experiments were performed in 5 groups. In each group, ten loops were used and the weight of a clot was statistically evaluated. The following items were investigated: 1) effect of LMWD on artificial clot formation, 2) effect of LMWD on thrombolysis of artificial thrombi, 3) effect of LMWD and combined use of UK on thrombolysis of artificial thrombi. It was varified that the blood containing 4.2mg of LMWD per ml is more significantly prophylactic for clot formation as compared with control (p<0.001, N=9). More 16.7mg of LMWD per ml showed significant thrombolytic activity compared with control (p<0.05, N=8). On thrombolytic activity, combined use of 15.4mg of LMWD per ml and 50 I. U. of UK per ml showed enhancement of 3.5 times over than control (p<0.01, N=8). Concomitant use of 3.8mg of LMWD per ml and 1, 000 I. U. per ml enhanced fibrinolytic activity 1.9 times than control (p<0.01, N=8).
The simultaneous combined administration of LMWD and UK will be more effective for thrombolytic treatment than UK only.

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Thrombolytic therapy with urokinase (UK) requires large quantities of UK to achieve the desired effect. In order to find a means of reducing the necessary quantity of UK, the effect of concomitant UK and low-molecular-weight dextran sulphate (DS) (3, 900) administration was examined in the rabbit. The fibrinolytic curves after DS administration moved to left and upward when compared to that of before DS administration, suggesting the increase of the sensitivity to UK. Such increase of the sensitivity to UK was reconfirmed in human.
The fibrinolytic activity in the euglobulin fraction after the concomitant administration of UK and DS was markedly enhanced and sustained for long periods after the completion of UK infusion, indicating that the inactivation rate constant of UK was smaller during the UK infusion.
These results indicate that concomitant administration of UK and DS may be helpful for reducing the necessary quantity of UK for thrombolytic therapy.

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The effects of urokinase (UK) on blood coagulation and fibrinolytic activities in 19 cases with cerebral infarction and 2 cases with myocardial infarction were evaluated.
Each case was administrated with 60, 000 or 480, 000 international unit of UK per day and this therapy was performed for one or seven days.
The following results were obtained.
1) The hyperfibrinolytic effects such as shortened euglobulin clot lysis time, decrease of plasminogen and antiplasmin were observed, but significant increased level of FDP was not always obtained.
2) The remarkable changes of prothrombin time, partial thromboplastin time, fibrinogen, α₁-antitrypsin and α₂-macroglobulin were not observed at the end of UK infusion.
3) The decrease of antithrombin III was shown in the cases administrated with high dose of UK.
4) It was necessary for detecting of the effects of UK on coagulofibrinolytic system to do the serial assay of these factors after UK infusion.
5) Most of the cases with excellent or good clinical course exhibited the remarkable increased level of FDP in the early stage of UK therapy or lack of the findings of suggesting hypercoagulability. The bleeding complications induced by UK therapy was not observed.
These results show that it is important to administrate the dose of UK enough to induce the increased level of FDP in early stage and to prevent the hypercoagulability by anticoagulant therapy when UK therapy was performed in the cases with cerebral infarction and myocardial infarction.

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The role of plasma fibrinogen in the formation of the stasis thrombus of the jugular vein and experimental thrombus of the femoral artery was studied in dogs. Marked hypofibrinogenemia was induced in the dogs 9 to 48 hours after the subcutaneous injection of defibrase, without any significant changes in platelet count and plasma coagulation factors except for fibrinogen. The score of the stasis thrombus was determined according to Wessler's standard. No thrombi were found in the jugular veins of five control dogs injected with normal saline alone. Thrombi were formed in all of the nine control dogs injected with 40ml of normal canine serum containing 20mg of watersoluble ellagic acid. In the eleven dogs with less than 65mg/dl of plasma fibrinogen concentration no thrombi developed following iv injection with serum and ellagic acid. Thrombi of score 2 to 4 were formed in eight dogs with fibrinogen concentration more than 120mg/dl after iv injection with serum and ellagic acid. In dogs with fibrinogen concentration 80-105mg/dl, thrombi of score 0-3 were observed.
The occluding thrombus was produced in all of control dogs on the chemically intimectomized femoral artery with stenosing ligature after iv injection with serum and ellagic acid. No thrombi developed in all of five defibrase injected dogs with fibrinogen concentration less than 60mg/dl on similary treated femoral artery following iv injection with serum and ellagic acid. Occluding thrombus was formed in 7 out of the 9 defibrase treateed dogs with fibrinogen concentration of 80-255mg/dl. From the results above described, it is suggested that plasma fibrinogen concentration more than 80mg/dl is essential for the formation of either stasis thrombus or experimental occluding arterial thrombus.

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It has been said that the incidence of postoperative deep vein thrombosis is significantly low in Japanese compared to Western peoples. The study was undertaken to investigate its incidence and risk factors in Japanese.
One hundred and twenty patients undergoing major abdominal surgery were investigated for deep vein thrombosis. All patients underwent postoperative isotopic scanning of the legs using the ¹²⁵I-fibrinogen technique. Systematic studies on blood coagulation and fibrinolysis were performed on the patients before and after operation.
In 15 of 120 patients there was the isotopic evidence of deep vein thrombosis, which was confirmed by ascending phlebography in ten in whom this procedure was performed. Thrombosis was unilateral in five and bilateral in ten. There were no physical signs suggesting thrombosis nor further propagation of thrombi into the popliteal vein or more proximal veins in any case, although seven of them recieved urokinase treatment immediately after detection of thrombosis in calf veins.
Deep vein thrombosis developed more frequently in patients older than 50 years and in female. Deep vein trombosis was mostly found in patients who had colorectal surgery. All except one had malignant disease. Ten of fifteen cases who developed deep vein thrombosis had complete study of blood coagulation and fibrinolysis showed definite hypercoagulability in thromboelastgram and increase of heparin tolerance compared to those who had no thrombosis. However, euglobulin lysis time was rather shortened, suggesting increased fibrinolysis.
The incidence of deep vein thrombosis in our series is definitely low compared to many reports in Western literatures. The cause of the low incidence remain open. Whatever the cause, the differences that we have demonstrated provide the first objective evidence that deep vein thrombosis is truely less frequent in Japanese than in Western peoples.

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The developmental stages of thrombi in 31 cases of clinically unsuspected DIC were morphologically classified. DIC was diagnosed by the presence of fibrin thrombi primarily in small vessels. Onset time of DIC was determined retrospectively.
Thrombi were divided into three major stage groups according to the presence of cellular reactions to them; thrombi without reaction of vascular endothelial cells or fibroblasts (stage I); those covered with vascular endothelial cells, either invaded or not, but without fibroblastic reaction (stage II), and those reacted by both (stage III).
Stages I and II were subdivided into two minor stage groups respectively, according to tinctorial properties of the thrombi for H. E., Picro-Mallory, PTAH, and PAS stains. Stage Ia denotes thrombi which stain weakly acidophilic with H. E. stain, red with Picro-Mallory stain and positive with PTAH, but negative with PAS stains; in stage Ib, thrombi are moderately acidophilic in H. E. stain, red in Picro-Mallory stain, positive in PTAH and PAS stains. In stage IIa, thrombi are more acidophilic in H. E. stain, red to reddish purple in Picro-Mallory stain, weakly positive in PTAH and positive in PAS stains. In stage IIb, they show strong acidophilic properties in H. E. stain, reddish to bluish purple in Picro-Mallory stain, red purple in PAS stain but no reactions to PTAH stain. Lastly, in stage III, they become granulation-like to scar-like in H. E., and blue in Picro-Mallory stains.
A close relation was noted between these morphological stages of thrombi and the duration of DIC; in less than 12 days, the thrombi were mostly in the stage I; between 13 to 30 days, they were in the stage I or II, and more than one month, in the stage I to III.
The above results suggest that this classification is useful for determination of the age of thrombi as well as for investigation of their natural history.

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Pulsed NMR (nuclear magnetic resonance) experiments of Carr-Purcell-Meiboom-Gill type have been performed on water flowing through a stenosis in a cylindrical tube which was made of acrylglass (with i. d. of 10mm and o. d. of 14mm). The constricted part of the stenosis is 7mm. Spin echo signals have been detected rf (radio frequency) phase-sensitively in a highly stabilized electromagnet with a linear magnetic field gradient. From measurements of the first spin echo amplitude, informations on velocity distribution are obtanined, while from those of the second spin echo informations on velocity fluctuation are obtained. When the magnetic field gradient vector is laid in the direction of the axis of the tube, one can observe the velocity component parallel to the axis. On the other hand, when the direction of the vector is perpendicular to the axis, one can observe the velocity component perpendicular to the axis. The relation between the spin echo signal and the amplitude of the field gradient has been observed behind the stenosis and compared with the case of a uniform cylindrical tube. The effect of disturbed flow behind the stenosis on spin echo signals are very large even at low Reynolds numbers (small flow rates). The velocity fluctuation ‹(δ_V)²›, the velocity component perpendicular to the axis, has been calculated at various flow rates from obtained date. Measurements have been done at flow rates from 6.0cm/s to 63.7cm/s.

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Blood supply to tissues through small vessels may be disturbed by arteriosclerotic narrowing. Recent hemorrheology have shown that erythrocytes could pass through the capillaries with smaller diameters than those of erythrocytes deforming themselves. Then, the clinical evaluation of erythrocyte deformability may be advantageous in the study on the circulation in arteriosclerotic disorders. The aim of this study is to elucidate the influences of the blood constituents on erythrocyte deformability and the difference in erythrocyte deformability between arterial and venous blood. Venous blood specimens were drawn from cubital veins in 61 subjects aged from 22 to 84 years, and arterial blood specimens were collected in 14 subjects from femoral arteries. Erythrocyte deformability was evaluated by the deformability index measure using filtration method developed by Reid et al. Freshly drawn whole blood anticoagulated with 3.8% sodium citrate was filtered through polycarbonate membrane with 5μ diameter pores at a pressure gradient of 20cm of water. Deformability index was expressed as the blood volume passed through the membrane in 1 minute. Readings were taken in triplicate on each sample. Mean values and standard deviations were as follows: 1.07±0.17ml/min in the twenties, 0.95±0.09ml/min in the thirties, 0.95±0.13ml/min in the forties, 0.95±0.25ml/min in the fifties, 0.82±0.12ml/min in the sixties, 0.16±0.17ml/min over seventy years old. The index in subjects aged over seventy was significantly lower than those in the other age groups. The index was directly proportional to blood ATP content, and inversely to fibrinogen, hematocrit and hemoglobin. The concentration of ATP in the blood passed through the membrane was higher than that of ATP measured before filtration. Neither deformability index nor ATP level indicated significant arteriovenous gap. Therefore, venous blood should be preferable to arterial blood in the determination of erythrocyte deformability. Decreased deformability index in the aged subjects may relate the accelerated formation of ischemic lesions in the tissues perfused by sclerotic arteries. The direct correlation between the index and ATP content, and high concentration of ATP in the blood passed through the membrane are in accordance with previous findings.
The close relations of the index to hemoglobin, hematocrit and fibrinogen indicate indirectly the contribution of erythrocyte deformability to blood viscosity as shown in previous papers. High blood viscosity promotes thrombus formation and may result in infarction. Therefore, in conclusion, the deformability index should be admitted as one of the useful measures in the observation of the aged subjects which might be accompanied with arteriosclerotic disorders.

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In a case of giant cavernous hemagioma of the liver, accompanied consumption coagulopathy and storage pool syndrome were treated successfully by the surgical resection of the tumor. This paper presents the electron microscopic observations of the tumor specimens, with regards to possible causative factors of these complications in the tumor.
Most abnormal vessels observed in the specimens are fairly small in size, ranging from 5μ to 35μ in diameter and consist of thin endothelial cells with partially rough and fluffy luminal surface. In some endothelial cells, phagocytosis of lipid-like substances and endothelial disruption are observed. An usual distinct basement membrane surrounding the whole vessel circumference is not evident, although discontinuous basement membrane is seen in a few vessels. Pericytes are rarely seen.
Erythrocytes, leukocytes and platelets are seen in the subendothelial or interstitial space outside the vessels, whether endothelial gap is observed or not. In these abnormal vessel lumina, increased platelets of different types such as almost normal, degranulated and aggregated, are observed, suggesting a sequestration of platelets. Platelet aggregates are seen more frequently in bifurcation of the vessels. Rarely, a platelet thrombus protrudes from the lumen to the subendothelial space through the endothelial gap. In these small vessels, thrombi consist of platelets and sometimes leukocytes, and fibrin is scarcely seen. In larger vessels, contrarily, thrombi consist mostly of fibrin.
Rouleau formation of erythrocytes is seen in some of the small vessels, and fragmentation of erythrocytes is observed near the fibrin clots in larger vessels.
The ultreastructure of giant cavernous hemangioma of the liver in this case may explain morphologically its causality on consumption coagulopathy and acquired storage pool syndrome.

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The prevention of postoperative intravascular coagulation is an important problem in surgery, but the definitive procedure has not been established. In 1974 the reduction of the deposition of fibrin in the lungs by Trasylol after intravenous injection of thromboplastin was reported by Diffang et al. That Gabexate mesilate (FOY), a synthetic protease inhibitor, weakly reduced the activity of thrombin is also known.
FOY could affect the prothrombin time and also the activated partial thromboplastin time at final concentration of 2.4×10^-5M, or more (Fig. 1). Therefore the deposition of fibrin in the lungs after injection of thromboplastin and thrombin was determined by the method of Saldeen (1969), in which ¹²⁵I-human fibrinogen was used. The rats were given 0.01% potassium iodine in their drinking water. One μCi radioactive human fibrinogen per 100g body weight was injected into a tail vein 48 hours before the experiment. Radioactivity in the rat's lung after bleeding was determined in the experiment. The rats were given 0.5mg of FOY one minute before thromboplastin or thrombin injection, and 5 minutes after the injection. The rats were killed 10 and 40 minutes after the beginning of the injection of thromboplastin, thrombin or saline. FOY injection was found to reduce the deposition of fibrin in the lungs significantly 10 minutes after the injection of 5mg of thromboplastin per 100g body weight by determining radioactivity in the lungs (Fig. 2). Meanwhile, FOY injection was not found to reduce the deposition of fibrin significantly 10 minutes after the injection of 12.5 units per 100g body weight (Fig. 3). However, the deposition of fibrin was somewhat reduced by FOY injection on the average as compared with the control. The alteration of plasma fibrinogen and serum FDP was determined 10 or 40 minutes after the injection of thromboplastin or thrombin. But there was no significant difference between two groups.
By microscope-search ten minutes after the injection of thromboplastin, moderate amounts of fibrin were observed in the lungs of the rat given thromboplastin alone, as well as in those injected with FOY and thromboplastin. There was no significant difference in the amounts of fibrin.
Finally, the present study suggests that a high concentration of FOY may be usefull as the inhibitor against the intravascular coagulation. That the activity of FOY was eliminated early after the injection, was due to a high activity of esterase in rats. From this property of FOY, it seems to be more appropriate to give this drug to patients after surgery rather than to ones with DIC.

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In Japan, gabexate mesilate (FOY) has been synthesized as a newly therapeutic agents for pancreatitis. It has been known that this protease inhibitor affects trypsin, plasmin, plasma kallikrein, thrombin and Cl-esterase. In this paper, the inhibitory effects of FOY on experimental DIC were investigated in comparison with those of aprotinin or heparin.
32 rabbits (matured male, body weight from 2.5 to 3.0kg) were used as experimental animals. These were divided into three groups. Each group was pretreated by infusion of FOY, aprotinin or heparin with 20mg/kg, 10, 000KIE/kg or 400 Units/kg respectively, and immediately after the infusion, thrombin, tissue thromboplastin or endotoxin were infused as DIC inducers. Several parameters (platelet counts, white blood cells, neutrophils, fibrinogen, FDP, platelet retention, platelet aggregation (ADP-induced), prothrombin time and partial thromboplastin time) were investigated before and after infusions of trigger substances. The changes of these parameters were expressed in accordance with the score system which had been ready beforehand, and then the inhibitory effects of these drugs on experimental DIC were examined statistically.
The results obtained are as follows;
1) Total scores of each group in thrombin-induced DIC were 31, 10, 24 and 21, in groups of control, FOY, aprotinin and heparin respectively. Scores in drug-pretreated group were significantly smaller than those in control group (P<0.05). In particular, the score of FOY was lower than those of the other groups statistically (p<0.05).
2) In tissue thromboplastin-induced DIC, scores were 35, 26, 24 and 20 in control, FOY, aprotinin and heparin each other, with no significant differences among the drug groups.
3) Scores in endotoxin-induced DIC were 39, 28, 29 and 31 in groups of control, FOY, aprotinin and heparin, respectively, showing no significant differences.
In conclusion, the results of the present study suggest that FOY was more effective than heparin or aprotinin on the inhibition of experimental DIC.

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Endotoxinemia was produced in six Japanese monkeys (7-13kg) by two suitably spaced (24hrs, Fig. 1, Experiment I) and by a continuous (8hrs, Fig. 1, Experiment II) intravascular injection with endotoxin (ET) from E. coli. The changes of concentration of ET in plasma, and faculties of coagulative, fibrinolytic, kinin forming and complement systems were investigated before and after treatment with ET. Body temperature, arterial pressure, pH, pO₂, pCO₂, Ht and Hb in blood were checked throughout the experiment.
The main observations were as follows.
[1] Two spaced injection of ET (0.3-2mg/kg, 3 monkeys)
Et in plasma rose to 0.1-1μg/ml of plasma immediately after the injection of ET and thereafter decreased gradually (Fig. 1, Experiment I). Activation of coagulation system was suggested by 60 to 70% decrease in platelet count and 20 to 30% prolongation of APTT and PT except for TT. Intrinsic coagulation factors, F. XII, F. XI, F. IX, F. VIII and extrinsic factor, F. VII decreased 20 to 30% after the treatment of first ET (ET-1), and further decreased 40 to 60% after second ET (ET-2). However, fibrinogen increased about two times before the treatment. Activation of fibrinolytic and kinin forming systems was indicated by the decrease of plasminogen and prekallikrein, generation of plasmin, kallikrein and FDP after ET-2 (Fig. 2). Complement factors, C₃proactivator, C₃c and C₄ were decreased after ET-1 and ET-2.
[2] Continuous injection of ET (1-5mg/kg/hr, 3 monkeys)
ET in plasma rose 1-10μg/ml of plasma immediately after the first injection of ET and maintained its concentration to the death (Fig. 1, Experiment II). Activation of coagulation system was suggested by the extreme decrease in platelet count, prolongation of APTT, PT and TT. The decreases of F. XII, F. X and F. II were observed at 3hrs after the treatment of first ET. The decreases of F. IX, F. VIII and F. V were observed at 6hrs, and F. VII and fibrinogen decreased at 9hrs (Table 1). These findings indicate that ET activates the intrinsic coagualtion system. Activation of fibrinolytic and kinin forming systems was observed markedly as compared as the experiment of two spaced injection of ET (Fig. 3). In complement system, alternative pathway was activated. In plasma inhibitors, antithrombin III decreased, but α₂-macroglobulin, C₁s inhibitor and α₂-plasmin inhibitor slightly decreased.
These results in Japanese monkeys treated with ET have led us to conclude that DIC produced with ET may have been occured initially by the activation of intrinsic coagulation factor, F. XII, accompanying by the activation of prekallikrein and plasminogen.

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The characteristics of the trigger substances of disseminated intravascular coagulation syndrome in the cases with gastric cancer and acute promyelocytic leukemia, and those of the serine proteases (Factor Xa, thrombin, plasmin and trypsin) were studies from the viewpoints of the prekallikrein activation.
The serine proteases and antithrombin III were purified by our methods, and prekallikrein was assayed by our method.
Results:
(1). Endotoxin (E. Coli, Difco Co., 0.2mg/ml in the final concentration) activated factor XII in the citrated plasma to show the shortened PTT, and the factor XIIa activated prekallikrein in the plasma. Kallikrein activated the purified factor VII from 25% to 47% compared with the normal plasma. Ten mg/ml of endotoxin in the plasma activated coagulation system to show 0.04units/ml of factor Xa and 0.025units/ml of thrombin assayed by the methods using synthetic chromogenic substrates.
(2). 6×10⁴/cmm of the cultured well differentiated adenocarcinoma cells and the pathologic cells in the case with acute promyelocytic leukemia activated 1μg of the purified bovine prekallikrein.
(3). Two units/ml of factor Xa, 20units/ml of plasmin and 0.03mg/ml of trypsin activated the prekallikreinat 10min activation time; Factor Xa: 25%, plasmin: 15%, and trypsin: 41%.
(4). The purified antithrombin III inhibited kallikrein, and heparin accelerated this inhibitory effect. Aprotinin (500units/ml in the final concentration) inhibited 85% prekallikrein activation by addition of kaolin. Dextran sulfate (0.3 and 3mg/ml in the final cocentration) showed the antithrombin activity, and inhibited prekallikrein activation by addition of kaolin; 0.3mg/ml: 78% and 3mg/ml: 64%.

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Nine out of 319 patients who underwent open heart surgery from March 1977 to July 1978 at the University Hospital developed the laboratory evidence of DIC and were administrated heparin. Among the 9 patients, 6 cases were tetralogy or pentalogy of Fallot. (27 of 319 patients were Fallot's disease.) Other cases were endocardial cushion defect, annuloaortic ectasia and ventricular septal defect with pulmonary hypertension.
Laboratory evidence of DIC was found within 3 days after surgery. They had many etiologic factors contributing to the production of DIC following cardiopulmonary bypass, i. e. a long term bypass, hemolysis, shock or low cardiac output, polycythemia and severe infection. Eight out of 9 patients were associated with shock or low cardiac output and 5 of them had a hematocrit greater than 50 per cent. We suggest that low cardiac output and polycythemia may be main factors to develop DIC.
When diagnosis of DIC was established, all patients had a bleeding tendeney. One patient had acute renal failure. During heparin therapy, 2 patients had renal dysfunction and one of them developed acute renal failure. In this series of 9 patients, 2 who had renal failure ultimately died. In other cases, there were recovery from the bleeding tendency and improvement in coagulation findings.
In coagulation studies, platelet count, prothrombin time, FDP and thrombelastograph were useful for diagnosis and judgement of recovery or progress of DIC.
Early diagnosis is important because early administration of heparin may inhibit progress of DIC and dysfunction of organ, such as renal failure.

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Immunoelectrophoretic studies were carried out on plasma samples of 5 young infants with idiopathic vitamin K deficiency and 5 infants with secondary vitamin K deficiency.
Abnormal prothrombin without procoagulant activity was demonstrated by one-dimensional immunoelectrophoresis (Laurell's method) and the Echis carinatus venom prothrombin assay in all of 10 patients with vitamin K deficiency in young infants.
The abnormal prothrombin was distinguished from normal prothrombin in lack of procoagulant activity, disability to bind to barium sulfate, and faster mobility in the presence of calcium ions. Thus, this abnormal prothrombin had the same characteristics as the abnormal prothrombin in coumarin-treated patients.
Abnormal factor IX without procoagulant activity was also demonstrated by the antibody neutralization technic in 7 patients.
In order to study the turnover of the abnormal prothrombin in those patients, two-dimensional immunoelectrophoresis was carried out serially after administration of vitamin K. Although the abnormal prothrombin gradually decreased concomitantly with increase of normal prothrombin after administration of vitamin K, the plasma samples from patients contained small amount of the abnormal prothrombin even in 48 hours after treatment. It was found that the abnormal prothrombin didn't change to normal prothrombin even in the presence of vitamin K in contrast with that in the liver.
Although the plasma of two patients with mild secondary vitamin K deficiency contained abnormal prothrombin as well as normal prothrombin, the plasma of 3 patients with hemorrhagic disease of newborn didn't contain abnormal prothrombin. Thus, the latter may not be only due to vitamin K deficiency, but also due to prematurity of the liver.

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The reptilase time of 61 patients with hepatic disease, i. e. chronic hepatitis (21 cases), cirrhosis (29 cases) and others (11 cases) was estimated by the modified method of reptilase R clotting time. Also relationship of level between the reptilase time and some such factors as the following was studied in hepatic diseases described above: Thrombin time, prothrombin time, FDP and various substances for the liver function test.
The results obtained are as follows.
1. Almost half number of patients with chronic hepatitis and cirrhosis revealed prolongation of the thrombin time. While, most cases of the same diseases described above showed prolongation of the reptilase time.
2. The patients with cirrhosis in whom the reptilase time was prolonged revealed relatively high levels of ZTT and GOT.
3. The patients with cirrhosis, showing high level of about 100 Karmen in GOT or 12 units in ZTT, has significantly prolonged reptilase and thrombin times.
4. A less correlation was found between the fibrinogen levels estimated by the clot weight method and thrombin or reptilase time in patients with chronic hepatitis and cirrhosis.
5. FDP was found in many cases of chronic hepatitis and cirrhosis, but not so much that the thrombin time or reptilase time was influenced by FDP.
It is concluded that a tendency of prolongation of the reptilase time is of value for screening for active form of cirrhosis.

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It has recently been shown by Fletcher and co-workers that circulating high molecular weight fibrinogen complexes are a most sensitive marker for detecting intravascular fibrin formation.
In the present investigation, circulating high molecular weight fibrinogen complexes were detected by gel chromatography of plasma and staphylococcal clumping test in 66 patients with various liver diseases. In normal adults, plasma high molecular weight fibrinogen complexes were lower than 0.5μg/ml.
In patients with liver diseases, the concentration of high molecular weight fibrinogen complexes in plasma elevated frequently.
Especially in patients with decompensated liver cirrhosis and hepatoma, the concentration of high molecular weight fibrinogen complexes elevated in most cases.
In patients with acute hepatitis, plasma high molecular weight fibrinogen complexes increased later than the elevation of GPT in serum.
Plasma high molecular weight fibrinogen complexes in patients with liver cirrhosis decreased markedly by administration of heparin.
From these results, it was suggested that subclinical intravascular coagulation exists frequently in patients with liver diseases.

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