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Objective: To observe the effect of Xiaoyu Zhixue tablet (XYZXT) on the expression of platelet membrane glycoproteins in patients with hemorrhagic thrombopathy, and to explore its possible mechanism. Methods; The total of 148 patients with hemorrhagic thrombopathy were randomly divided into two groups, the traditional Chinese medicicne (TCM) group (n= 98) treated with XYZXT and the Western medicine (WM) group (n= 50) treated with adrenosin, vitamins C, K and P, both for 6 months. The therapeutic effect and the recovery rate of platelet aggregation in the two groups were observed. And platelet membrane glycoprotein (GP) Ib/IX, GPII b/IIIa complexes, GPIb, GPIIb, GPIIIa and P-selectin were analyzed by flow cytometry in both groups before and after treatment and also in 34 normal healthy subjects.Results: The total effective rate of hemostasis was 89.8% in TCM group and 54.0% in the WM group (X2=45.83,P<0.01), and the recovery rate of platelet aggregation was 72.4% and 4.0% respectively (X2= 62.06,P<0.01). The fluorescence intensity of GP I b/IX, GP II b/III a complexes, GPIb, GP IIIa and P-selectin were lower in both groups before treatment than those in the healthy subjects. Expression of above-mentioned marks was elevated in TCM group after 6 months’ therapy, which was insignificantly different as compared with the healthy subjects (P>0.05) and higher than those in the WM group (P<0.05).Conclusion: One of the mechanisms in treating hemorrhagic thrombopathy with XYZXT is that it could regulate the expression of GP I b/IX, GPII b/IIIa complexes, GP I b, GPIIIa and P-selectin at the level of receptor protein.

Summary: The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular α-granule thrombospondin (TSP) were examined and the inhibition of β-thromboglobulin (β-TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080± 17010 molecules/platelet as compared with 13190±4810 from the controls (P< 0.01). No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III,(P> 0.05). The GPIV redistribution on active platelet membrane induced thrombin (IU/ml) from CML and healthy donors was 44320 ±32310 and 22800± 12700 molecules/platelet respectively (P< 0.01). The difference in the release of intracellular α-granule TSP between CML and the control group was not found (P> 0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high levels of β-TG and PF4 in sera inhibited release of intracellular α-granule TSPin vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of internal TSP pools is hindered by either β-TG or PF4 in sera.

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular α-granule thrombospondin (TSP) were examined and the inhibition of β-thromboglobulin (β-TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080± 17010 molecules/platelet as compared with 13190±4810 from the controls (P< 0.01). No abnormality was found in the distribution of platelet membrane GPIband GPIIb/III,(P> 0.05). The GPIV redistribution on active platelet membrane induced thrombin (IU/ml) from CML and healthy donors was 44320 ±32310 and 22800± 12700 molecules/platelet respectively (P< 0.01). The difference in the release of intracellular α-granule TSP between CML and the control group was not found (P> 0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high levels of β-TG and PF4 in sera inhibited release of intracellular α-granule TSPin vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of internal TSP pools is hindered by either β-TG or PF4 in sera.

Summary: We applied an indirect hemagglutination inhibition test to detect platelet-associated immunoglobulin G (PAIgG). This test is relatively easy to perform and has good reproducibility. The PAIgG value detected with this method in 45 normal donors was 0–5 fg per platelet. In our series there were 15 ITP patients, of whom two had been in complete remission and had normal PAIgG values. In the remaining 13 cases, 11 were found to have increased PAIgG values (more than 5 fg per platelet). In cases where corticosteroids or splenectomy were effective, rising of the platelet count was associated with PAIgG waning. Preliminary analysis of clinical data of ITP patients suggested that the clinically diagnosed ITP might be of heterogeneous pathogenesis. A further study is warranted.

Summary: This report presents 37 cases of acute leukemias, showing that the skin-window cellular response was markedly reduced. Immature leukemic cells were not found in the exudates. Acellular response and “delayed” cellular response should be of a prognostic significance. Chemotherapeutic agents and adrenocorticosteroid do rot per se depress the cellular response and phagocytic ability in induced inflammatory exudates.

Summary: Growth pattern in vitro and karyotype of cultured bone marrow cells were studied in 31 patients with acute nonlymphocytic leukemia. Two different growth patterns were identified: cluster growth (pattern I), and no growth (pattern ll). Patients with growth pattern I had a significantly higher complete remission (CR) rate clinically than those with growth pattern I. A serial study of 23 patients during induction treatment showed three different patterns: (1) colony growth, (2) persistence of clusters, and (3) persistence of no growth. The prognosis of the patients with the first pattern was most favourable, 11 of 13 patients achieved CR on an average of 24 days after first detection of colony formation. The data from 152 cultures showed that the colony forming ability had a negative correlation with the percentage of marrow blasts. Karyotype analysis with G-banding technique was successfully performed in 26 patients, in 14 of whom abnormal clones were detected. There was no difference in CR rates between groups NN, A A and AN, although CR was more easily achieved in patients with t(8; 2l). No relationship was seen between growth pattern and karyotype.

The effects of intensive versus regular therapy on incidence and progress of microalbuminuria in type 2 diabetes were compared. During a follow-up of 3 years, 96 cases of diabetes mellitus were randomized to intensive and regular therapy groups. HbA1cgoal was same in the two groups, but the goal of blood pressure (Bp) and lipid was more strict in the intensive therapy group than in the regular therapy group. There was statistically significant difference in the incidence and progression of vascular complications between the two groups. Logistic stepwise-regression analysis (odds ration, OR) showed that there was significant difference in the progression of nephropathy (OR 0.24, 95% CI 0.12–0.76), retinopathy (OR 0.38, 95% CI 0.16–0.88), peripheral neuropathy (OR 0.42, 95% CI 0.22–0.86) and autonomic neuropathy (OR 0.29, 95% CI 0.12–0.86) between the two groups (P<0.01). It was concluded that intensive blood glucose controlling could retard diabetic vascular complications. Intensive therapy of multiple factors interventions (controlling Bp, regulating blood lipid, improving microcirculation) could decrease various risk factors for diabetic vascular complications.

Summary: The changes of platelet fibrinogen in patients with obscure cutaneous mucosal bleeding and their relation with disease were evaluated. By using flow cytometry and Western blot, the platelet binding site of fibrinogen and the expression of platelet fibrinogen in 8 cases of obscure bleeding were detected. The results showed that platelet surface binding sites of fibrinogen were 65.38±3.62 in the resting state and 65.25±5.78 after activated by adenosine diphosphate, showing no significant difference as compared with control group (P>0.05). The expression of platelet fibrinogen was obviously decreased in 4 patients and increased in one case as compared with control group. It was concluded that the binding sites of fibrinogen on the platelet surface were normal in the patients with idiopathic bleeding platelet disorder. The abnormal of platelet fibrinogen might be one of hemorrhagic reasons in partial patients.

The changes of platelet fibrinogen in patients with obscure cutaneous mucosal bleeding and their relation with disease were evaluated. By using flow cytometry and Western blot, the platelet binding site of fibrinogen and the expression of platelet fibrinogen in 8 cases of obscure bleeding were detected. The results showed that platelet surface binding sites of fibrinogen were 65.38±3.62 in the resting state and 65.25±5.78 after activated by adenosine diphosphate, showing no significant difference as compared with control group (P>0.05). The expression of platelet fibrinogen was obviously decreased in 4 patients and increased in one case as compared with control group. It was concluded that the binding sites of fibrinogen on the platelet surface were normal in the patients with idiopathic bleeding platelet disorder. The abnormal of platelet fibrinogen might be one of hemorrhagic reasons in partial patients.

Summary: Using the method of dual-wavelength measurement of platelet [Ca2+]i and Fura-2 as the Ca2+ fluorophore probe, we measured the effect of acidic Mucopolysaccharide fromSticopus Japonicus Selenka (SJAMP) on platelet [Ca2+]i. The results showed that the most significant increase in platelets [Ca2+]i was seen when the concentration of SJAMP was 100μg/ml and the elevation of normal platelet [Ca2+]i was 93.96 ± 10.24 nmol/L (n=10). In the presence of extracellular Ca2+ (1 mmol/L), the magnitude of platelet [Ca2+]i response to SJAMP was increased and the [Ca2+]i could reach 116.72±10. 66 nmol/L (n = 10). On the other hand, the magnitude of increased platelet [Ca2+]i induced by SJAMP was smaller and the duration of [Ca2+]i reaching the highest level was longer when compared with other platelet aggregation agents. In the mean time, if platelets were first incubated with cyclooxygenase inhibitor, the rise of [Ca2+]i evoked by SJAMP was inhibited. The results indicated that the mechanism of the rise of [Ca2+]i induced by SJAMP might be dependent upon the generation of prostaglandin endoperoxides and(or) TXA2.

The aggregation and ATP release of placelet of normal subjects were measured by platelet lumi-aggregometer. It was found that the aggregation curve induced by SJAMP at the concentration of 100 mg/L was a typical second phase aggregation. There existed a certain lag between platelet aggregation and secretion. The secretion actually began slightly after the second phase of aggregation, suggesting that the second phase aggregation induced by SJAMP is not dependent upon the release of contents of dense granule alone. If platelets were incubated with cyclo-oxygenase inhibitor, the second phase aggregation was inhibited and no ATP was released. The results indicated that the aggregation and release reaction induced by SJAMP were dependent upon the generation of prostaglandin endoperoxides and TXA2in normal subjects. The amount of ATP release was 0. 69 ±0. 22 nmol/108platelets as stimulated with SJAMP (100 mg/L). But the amount of ATP release were 1. 60±0. 25 and 1. 37±0. 15 nmol/108platelets when platelets were stimulated with ADP (5 ώmol/L) and collagen (5 mg/L). The amount of ATP release induced by SJAMP was significantly lower than that of ADP and collagen. These findings indicated that SJAMP was a weaker agonist than ADP in terms of platelets release reaction.

Using the method of dual-wavelength measurement of platelet [Ca2+]iand Fura-2 as the Ca2+fluorophore probe, we measured the effect of acidic Mucopolysaccharide fromSticopus Japonicus Selenka(SJAMP) on platelet [Ca2+]i. The results showed that the most significant increase in platelets [Ca2+]iwas seen when the concentration of SJAMP was 100μg/ml and the elevation of normal platelet [Ca2+]iwas 93.96 ± 10.24 nmol/L (n=10). In the presence of extracellular Ca2+(1 mmol/L), the magnitude of platelet [Ca2+]iresponse to SJAMP was increased and the [Ca2+]icould reach 116.72±10. 66 nmol/L (n= 10). On the other hand, the magnitude of increased platelet [Ca2+]iinduced by SJAMP was smaller and the duration of [Ca2+]ireaching the highest level was longer when compared with other platelet aggregation agents. In the mean time, if platelets were first incubated with cyclooxygenase inhibitor, the rise of [Ca2+]ievoked by SJAMP was inhibited. The results indicated that the mechanism of the rise of [Ca2+]iinduced by SJAMP might be dependent upon the generation of prostaglandin endoperoxides and(or) TXA2.

Summary: The aggregation and ATP release of placelet of normal subjects were measured by platelet lumi-aggregometer. It was found that the aggregation curve induced by SJAMP at the concentration of 100 mg/L was a typical second phase aggregation. There existed a certain lag between platelet aggregation and secretion. The secretion actually began slightly after the second phase of aggregation, suggesting that the second phase aggregation induced by SJAMP is not dependent upon the release of contents of dense granule alone. If platelets were incubated with cyclo-oxygenase inhibitor, the second phase aggregation was inhibited and no ATP was released. The results indicated that the aggregation and release reaction induced by SJAMP were dependent upon the generation of prostaglandin endoperoxides and TXA2 in normal subjects. The amount of ATP release was 0. 69 ±0. 22 nmol/108 platelets as stimulated with SJAMP (100 mg/L). But the amount of ATP release were 1. 60±0. 25 and 1. 37±0. 15 nmol/108 platelets when platelets were stimulated with ADP (5 ώmol/L) and collagen (5 mg/L). The amount of ATP release induced by SJAMP was significantly lower than that of ADP and collagen. These findings indicated that SJAMP was a weaker agonist than ADP in terms of platelets release reaction.

Male rats at six weeks of age were divided into 5 groups at random: in group 1, the rats with diabetes received 70 % pancreatectomy; group II had sham-operation serving as controls; diabetic rats in group III were treated with phloridzin; In group IV rats received sham-operation and phloridzin treatment and group V were phloridzin-treated diabetic rats to be studied after discontinuance of phloridzin. 70 days after surgery, the weights and insulin contents of operated remnant pancreas were markedly higher than the expected value of 30 %, reaching 44 % (48. 2 %±15. 2 %), demonstrating that the remnant pancreas still had capacities of compensatory regeneration and proliferation capacities. Phloridzintreated diabetic rats completely returned to normal in terms of oral glucose tolerance and insulin sensitivity. Discontinuation of phloridzin treatment in diabetic rats resulted in the recurrence of insulin resistance. These results suggested that normalization of hyperglycemia could ameliorate insulin resistance under diabetic conditions.

Summary: Male rats at six weeks of age were divided into 5 groups at random: in group 1, the rats with diabetes received 70 % pancreatectomy; group II had sham-operation serving as controls; diabetic rats in group III were treated with phloridzin; In group IV rats received sham-operation and phloridzin treatment and group V were phloridzin-treated diabetic rats to be studied after discontinuance of phloridzin. 70 days after surgery, the weights and insulin contents of operated remnant pancreas were markedly higher than the expected value of 30 %, reaching 44 % (48. 2 %±15. 2 %), demonstrating that the remnant pancreas still had capacities of compensatory regeneration and proliferation capacities. Phloridzintreated diabetic rats completely returned to normal in terms of oral glucose tolerance and insulin sensitivity. Discontinuation of phloridzin treatment in diabetic rats resulted in the recurrence of insulin resistance. These results suggested that normalization of hyperglycemia could ameliorate insulin resistance under diabetic conditions.

Summary: The authors present a study of the growth of fibroblast colony forming units in tone marrow of normal adults and untreated patients with acute nonlymphocytic leukemia. The culture conditions and cytomorphology are also discussed.