Effect of Hanfangji compound on proliferation of hepatic stellate cells and expressions of TTR, ITIH1 and SERPINF2 / 中国中药杂志

<p><b>OBJECTIVE</b>To observe and compare the effects of Hanfangji Compound and IFN-gamma on expressions of transthyretin (TTR) , inter-alpha inhibitor H1 (ITIH1) and serpin peptidase inhibitor clade F member 2 (SERPINF2) of hepatic stellate cells (HSC-T6).</p><p><b>METHOD</b>Hanfangji Compound and IFN-gammaof different concentrations were used in hepatic stellate cell-T6 (HSC-T6) for 48 h. Flow cytometer was used to detect the effects of Hanfangji Compound and IFN-gamma on HSC proliferation. RT-PCR method was adopted to detect mRNA expressions of TFR, ITIH1 and SERPINF2. TTR, ITIH1 and SERPINF2 secretions were detected by ELISA. The protein localizations of TTR, ITIH1 and SERPINF2 were examined by immune fluorescence. The protein expression of TfR and ITIHI were determined by Western blot.</p><p><b>RESULT</b>After Hanfangji Compound and IFN-gamma were adopted in HSC-T6, compared with the control group, the cell proliferation was inhibited obviously (P < 0. 05) , protein expressions of TTR, ITIH1 and SERPINF2 and mRNA expression increased significantly, with certain correlation with concentrations of Hanfangji Compound. The 2. 5 g L-I Hanfangji Compound group was superior to the IFN-gamma group (P <0. 05).</p><p><b>CONCLUSION</b>Hanfangji Compound can inhibit HSC proliferation, upregulated TTR, ITIH1 and SERPINF2 proteins and mRNA expression, which may be one of mechanisms of anti-hepatic fibrosis of Hanfangji Compound.</p>

Novel distribution pattern of fibrinolytic components in rabbit tissues extract: a preliminary study / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>The purpose of this work was to investigate the distribution pattern of fibrinolytic factors and their inhibitors in rabbit tissues.</p><p><b>METHODS</b>The components of the fibrinolytic system in extracts from a variety of rabbit tissues, including tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and alpha(2) plasmin inhibitor (alpha(2)PI), were determined by colorimetric assay.</p><p><b>RESULTS</b>The tissue extracts in renal, small intestine, lung, brain and spleen demonstrated strong fibrinolytic function, in which high activity of tPA, Plg and Pl was manifested; whereas in skeletal muscle, tongue and stomach, higher activity of PAI-1 and alpha(2)PI showed obviously. Also excellent linear correlations were found between levels of tPA and PAI-1, Pl and alpha(2)PI, Plg and Pl. In related tissues, renal cortex and renal marrow showed distinctly higher activity of tPA and lower activity of PAI-1, with the levels of Plg and Pl in renal cortex being higher than those in renal marrow, where the alpha(2)PI level was higher than that in renal cortex. Similarly, the levels of tPA, Plg and Pl in small intestine were higher than those in large intestine, but with respect to PAI-1 and alpha(2)PI, the matter was reverse. In addition, the fibrinolytic activity in muscle tissue was lower, however, the levels of tPA, Plg, and Pl in cardiac muscle were obviously higher than those in skeletal muscles, and the levels of PAI-1 and alpha(2)PI were significantly lower than those in skeletal muscle.</p><p><b>CONCLUSION</b>Our data demonstrate that a remarkable difference of the fibrinolytic patterns exists in rabbit tissues, which has probable profound significance in understanding the relationship between the function of haemostasis or thrombosis and the physiologic function in tissues.</p>

Inherited Thrombophilia

Inherited thrombophilia has been an area of intense clinical research since the description of deficiency of antithrombin III [1965], protein C [1981] and protein S [1985]. It is now possible to identify the inherited thrombophila in about 50% of cases of venous thrombosis. Certain components of inherited thrombophilia have a major impact on anticoagulant therapy, so basic knowledge about them is necessary in every medical speciality

Plasma markers of thrombin generation, fibrinolysis and activated platelets in sickle cell disease

Thrombotic phenomenon has an important role in the vasooclusive manifestations of sickle cell disease [SCD] that dominate its clinical spectrum. For evaluation of thrombotic activities in SCD patients, the markers of thrombin generation: thrombin anti-thrombin complex [TAT] and prothrombin fragment 1, 2 [F 1, 2], enhanced fibrinolysis markers: plasmin anti- plasmin complex [PAP] and D-dimer and platelet activation markers: platelet factor 4 [PF4] and B - thromboglobulin [BTG] were measured in 35 patients with SCD [25 during steady state and 10 during crisis], compared to 12 normal controls. The measurements were performed by enzyme-linked immunosorbent assays. The thrombotic markers [TAT and F 1, 2], fibrinolytic markers [PAP and D-dimer] and platelet activation markers [PF4 and BTG] were significantly increased in SCD patients during steady state. During vasooclusive crisis, there were marked increase in TAT, D- dimer, BTG and PF4 while there was no significant increase in the levels of PAP and F 1, 2. Also, there was significantly positive correlation among thrombin generation markers, platelet activation markers and fibrinolysis markers in SCD patients during steady state. During episodes there was significant positive correlation among markers of thrombin generation, fibrinolysis and platelet activation except PAP and F 1, 2 markers as compared with asymptomatic intervals. We concluded that, during pain episodes, there was enhanced platelet procoagulant activity, elevated fibrinolytic activity and thrombin generation. These changes may predict the frequency of pain. These findings suggested that increased their levels may increase the risk of thrombosis in these patients and establish a relationship between the laboratory determination of these parameters and clinical pain episodes in patients with SCD. The findings of platelet activation, fibrinolytic activity and thrombin generation in asymptomatic patients indicate ongoing subclinical thrombogeinc activity in patients with SCD

Purification of plasmin-alpha2-antiplasmin complex by lysine-sepharose 4B affinity chromatography employing sequential elutions / 中国实验血液学杂志

To easily obtain plasmin-alpha(2)-antiplasmin complex (PAP) with high purity, the purification procedure was improved by authors. After urokinase activated fresh human platelet-deficient plasma had been applied to Lysine-Sepharose 4B affinity chromatography column, elutions were sequentially performed by several eluents with different ingredients. The results showed that 3.0 mg PAP could harvested from 100 ml fresh plasma by this method and the whole procedure could be finished within several hours. In conclusion, this procedure is simple, rapid, economical and high-yield.

Hypercoagulable state in children with nephrotic syndrome: a study of some markers of hemostasis activation

This study included 50 children with nephrotic syndrome; 28 were in relapse [group I] and 22 were in therapy withdrawal after successful remission [group II]. Twenty healthy children were included as controls. Thrombin antithrombin complex [TAT], fibrin monomer [FM], plasma fibrinogen, alpha-2-antiplasmin [alpha-2AP] tests and platelet counts were performed to detect the coagulation anomalies that are suggestive of a hypercoagulable state in the nephrotic cases. Serum albumin, total cholesterol and triglycerides levels were also determined. In conclusion, data suggested the presence of a prethrombotic state in the nephrotic cases, particularly in the relapse group. Elevated TAT, FM as well as fibrinogen and platelet count seem to be reliable indicators of this prethrombotic state. A significant reduction in the anti-fibrinolytic marker alpha-2AP was detected in the plasma of relapsing nephrotic cases

A Case of Protein S Deficiency with Cerebral Infarction / 대한소아혈액종양학회지

Certain natural anticoagulants are known to play critical roles on the control of hemostasis in addition to the physiologic role of fibrinolysis. Major antiproteases of this group include antithrombin III, protein C, protein S, alpha-2-antiplasmin, alpha-2-macroglobulin, etc. There has been increasing recognition of the importance of the control of coagulation and fibrinolysis, both by certain physiologic processes and by natural inhibitors. Especially, activated protein C (protein Ca) and its cofactor, protein S, now are recognized as significant modulators of fibrinolysis, thrombosis and hemostasis. Children and adults with decreased levels of either protein are at increased risk from thrombosis. We experienced a 5 year-old girl patient with cerebral infarction who initially presented with abnormal movement of left extremities. She had lacunar infarction in the posterior limb of the right internal capsule and hypoplastic left carotid artery on brain MRI and MR angiography. She was diagnosed to have protein S deficiency with the level of 43% in the screening test of hypercoagulable state. She remains clinically well after heparin and warfarin treatment with regular follow-up of prothrombin time. A brief review of the literature ensues with the case report.

The Role of Thrombophilia and Hypofibrinolysis in Legg-Calv-Perthes Disease / 대한정형외과학회잡지

PURPOSE: The object of present study was to investigate the relationship between LCPD and the abnormality of certain plasma proteins affecting clot mechanism and fibrinolysis in patients with LCP disease. MATERIALS AND METHODS: Twenty-five consecutive patients who had been diagnosed as LCP disease were matched with twenty-five controls for gender, age (2-year range) , and the time of presentation (1-year range) . There were twenty-three boys and two girls. The mean age of the children when the LCP disease was diagnosed was 6.7 years ( range, 2.1-12.8 years) , and the mean age at the time of the present study was 7.9 years ( range, 3.4 - 13 year) . Thrombotic disorders were investigated for protein C activity/antigenicity, protein S activity/antigenicity, antithrombin III, anticardiolipin antibody Ig G, anticardiolipin antibody Ig M, lupus antibody. Fibrinolytic disorders were investigated for tissue type plasminogen activator (t-PA) , plasminogen, plasminogen activator inhibitor-1 (PAI-1) , alpha 2 plasmin inhibitor, lipoprotein (a) . Wilcoxon rank sum test was used for the comparison. RESULTS: There was no significant difference in coagulation system and fibrinolytic system between patients and controls. CONCLUSION: Our results suggest that abnormality in coagulation and fibrinolytic system is not associated with Legg-Calv -Perthes disease.

Effects of Intermittent Sequential Pneumatic Compression on Coagulation and Fibrinolysis in Multiple Trauma

BACKGROUND: After multiple trauma, blood coagulation activity is enhanced and fibrinolytic activity is suppressed by overproduction of plasminogen activator inhibitor-1 (PAI-1). Intermittent sequential pneumatic compression (ISPC) is an effective method to prevent deep vein thrombosis. Its action is explained by the mechanical effect on blood flow, as well as by the enhancement of fibrinolysis by the reduction of PAI-1. The aim of this study was to determine the effects of ISPC on coagulation and fibrinolysis after multiple trauma. METHODS: Thirty-nine trauma patients were either treated with ISPC (ISPC group, 20 patients) or without ISPC (control group, 19 patients). We measured the plasma levels of the thrombin antithrombin III complex (TAT), the plasmin alpha 2 plasmin inhibitor complex (PIC), the tissue plasminogen activator (t-PA), and the plasminogen activator inhibitor-1 (PAI-1) on admission and at 1, 2, 3, 6, 12, and 24 hours after admission. RESULTS: The TAT was higher than normal in both groups, with no significant difference between the two groups throughout the study period. The PIC level of ISPC group was significantly higher than that of the control group. In the ISPC group, the PIC level increased gradually, reaching a peak at 3 hours and decreasing thereafter. In the control group, the PIC level increased to a peak level at 2 hours. The TAT/PIC ratio dropped in the first two hours and increased at 3 hours, dropping again thereafter. In the ISPC group, the ratio dropped gradually without an intermittent fluctuation. At 3 and 6 hours, the control group showed a significantly greater ratio compared to the ISPC group. PAI-1 was higher than normal in bothgroups, with a significantly lower level in the ISPC group from 2 hours to 24 hours. For the t-PA level, no difference was noted between the two groups, with the peak level occurring at 1 hour. The PAI-1/t-PA ratio was significantly greater in the control group from 2 hours to 12 hours than in the ISPC group, but the difference was not significant at 24 hours. CONCLUSIONS: In multiple trauma patients, ISPC does not seem to affect coagulation, but enhances fibrinolysis through suppressed PAI-1 production. This effect of ISPC may be maintained for 12 hours.

Alteration of Coagulation and Fibrinolysis in Multiple Trauma

BACKGROUND: After multiple trauma, blood coagulation activity is enhanced and fibrinolytic activity is suppressed. Due to high tissue thromboplastin concentration in cerebral tissue, more serious coagulation and fibrinolytic abnormalities may occur when concomitant head trauma is present. The aim of this study was to determine the changes in coagulation and fibrinolysis after trauma and the effects of head trauma on coagulation and fibrinolysis. METHODS: This study includes 35 trauma patients: 16 patients with head trauma (group A) and 19 patients without head trauma (group B). We measured the plasma levels of functional protein C, antithrombin III (AT III), thrombin antithrombin III complex (TAT), plasmin alpha 2 plasmin inhibitor complex (PIC), tissue plasminogen activator antigen (t-PA), and plasminogen activator inhibitor-1 antigen (PAI-1) on admission and on days 1, 2, 4, and 6 after the trauma. RESULTS: The TAT and the TAT/PIC were significantly higher in group A than in group B on all days. PIC was significantly lower in group A than in group B on all days except the day of admission. Over the course of time, the TAT and the TAT/PIC decreased in both groups and PIC increased. On admission, the PAI-1 of both groups was increased, but it decreased over the course of time. The t-PA was increased on admission, was suppressed on the 1st day, and then increased again. The PAI-1 and the t-PA showed no significant difference between the two groups. CONCLUSIONS: After multiple trauma, coagulation was enhanced and fibrinolysis was suppressed. Enhanced coagulation and suppressed fibrinolysis were significantly greater in group A than in group B.

Interaction of bovine alpha2 macroglobulin with proteinases

Alpha2, Macroglobulina, uma proteína inibidora de proteases, foi isolada do plasma bovino. O processo de purificaçäo foi monitorado por imunodifusäo e imunoeletroforese cruzada com soro anti alpha2M-humana. Para as determinaçöes quantitativas foi preparado um soro anti alpha2M bovino. A preparaçäo mais pura de alpha2M foi obtida por cromatografia de afinidade e usada como padräo primário na imunodifusäo radial de Mancini. Preparaçöes de alpha2M foram usadas em testes de ligaçäo com tripsina e plasmina (tituladas com NPGB). Nos testes de ligaçäo 50 por cento de plasmina e 35 por cento de tripsina foram "protegidas" pela alpha2M. Näo foi possível determinar se houve incidência na ligaçäo ou se a perda de atividade ocorreu por alteraçöes na afinidade do complexo alpha2M-protease, em relaçäo aos substratos usados

Alpha2-Antiplasmin deficiency as a risk factor for bleeding from esophageal varices

The coagulation state and alpha [2] antiplasmin level and their relation to the degree of hepatic dysfunction were evaluated in 16 patients with bleeding esophageal varices and 10 healthy controls. Prothrombin time [P.T.], partial thromboplastin time [P.T.T.] and thrombin clotting time were significantly prolonged in bleeders compared to controls [P<0.01], P.T.T prolongation was more pronounced in ascitic than non-ascitic bleeders. Fibrinogen was normal in both bleeders and controls but its level was significantly lower in ascitic than non-ascitic bleeders [P<0.01]. Alpha [2] antiplasmin level was significantly lower in bleeders than controls [P<0.01] and within the bleeder group, it was significantly lower in ascitic than non-ascitic bleeders [P<0.01]. This low level was associated with positive fibrin degradation products and normal platelet count in 6 patients [4 with ascites] pointing to its responsibility for the hyperfibrinolytic state occurring in such patients. Alpha [2] antiplasmin level and fibrinogen were inversely correlated with liver function tests [SGOT, SGPT, alkaline phosphatase and total bilirubin]. It is concluded that hepatic dysfunction adversely affects blood coagulation resulting in aggravation of bleeding from esophageal varices. The decrease of synthesis of alpha [2] antiplasmin may be responsible for aggravating bleeding. When associated with positive FDP's and normal platelet count, antifibrinolytic agents may be considered as potential therapeutic agents

Diagnostic and prognostic significance of antithrombin III, plasminogen and alpha-2-artiphasmin in jaundiced patients

This study was designed to find out the diagnostic and prognostic usefulness of Antithrombin-III, [AI-III] Plasminogen and Alpha-2 antiplasmin in different types of jaundice. AT-Ill, plasminogen and alpha-2 antiplasmin were studied in 43 jaundiced patients [Hepatic metastasis n = 9 and Hepatocel lular, n = 17, and obstructive, n = 17] and compared to 10 healthy controls. The mean serum levels of alpha-2 antiplasmin, AT-III and plasminogen were significantly decreased in hepatic metastasis and hepatocellular subgroups compared to control and cholestatic subjects. No significant differences were noticed between control subjects and cholestatic patients except for significant elevation in serum level of alpha-2 antiplasmin in the latter. In hepatocelluar jaundiced patients, these parameters were markedly decreased in fulminant hepatitis patients compared to cirrhotics and patients with acute viral hepatitis. Also in Hepatocellular group, a significant negative correlations could be elicited between bilirubin [Total and direct and SGOT on one side and AT-Ill and plasminogen on the other side. Also significant negative correlations were noticed between alpha-2 antiplasmin and each of total bilirubin and SGOT. It can be concluded that alpha-2 antiplasmin, AT-Ill and plasminogen may play a role in etiological diagnosis as well as prognostic follow up of jaundiced patients