摘要
Objective: To compare the safety and efficacy of bivalirudin versus unfractionated heparin during perioperative period of percutaneous coronary intervention(PCI) in real-world. Methods: A total of 13 097 serial patients who underwent PCI from January 2016 to November 2018 in the Northern Theater Command were enrolled in the present study. Patients were stratified as the bivalirudin group or the heparin group according to antithrombotic therapy during PCI. The primary efficiency endpoint was 30-day net adverse clinical event(NACE), defined as all-cause death, re-infarction, urgent target lesion revascularization (uTLR), stroke or any bleeding. The second efficiency endpoint was 30-day major cardiac and cerebral events (MACCE), defined as all-cause death, re-infarction, uTLR and stroke. Additional end points included the rates of stent thrombosis at 30 days. Propensity scores included clinical and demographic variables, with 1∶2 matching. Compared the incidence of events above between the two groups before and after matching. Results: Among the 13 097 included patients(age was (61±10) years old), 3 421 (26.1%) were female. And 2 734 patients were divided into the bivalirudin group, and 10 363 patients to the heparin group(5 468 after matching). Before propensity score matching, patients in bivalirudin group were older and received higher levels of CRUSADE score than heparin group. These patients were more likely to have hypertension and more with ST-segment elevation acute coronary syndromes(all P<0.05). After propensity score matching, the incidence of 30-day NACE(3.8%(103/2 734) vs.5.0%(271/5 468), P=0.015) and any bleeding (2.0%(54/2 734) vs. 2.8%(151/5 468), P=0.032) in the bivalirudin group were lower than that in the heparin group, but the incidence of MACCE (1.9%(51/2 734) vs. 2.3%(127/5 468), P=0.180) and stent thrombosis (0.1%(2/2 734) vs. 0.1%(3/5 468), P=1.000) were comparable between the two groups. Conclusion: The risk of bleeding and the incidence of NACE are significantly lower for patients using bivalirudin during perioperative period of PCI compared to heparin, without significant differences in ischemic events.
Subject(s)
Aged , Female , Humans , Middle Aged , Anticoagulants/therapeutic use , Heparin/therapeutic use , Hirudins , Peptide Fragments , Percutaneous Coronary Intervention , Perioperative Period , Recombinant Proteins , Treatment Outcome摘要
Abstract Purpose: To investigate whether hirudin exerts its antithrombin action to decrease the ratio of Human Microvascular Endothelial Cells (HMVECs) apoptosis. Methods: Human microvascular endothelial cells (HMVECs) cultured in the third and fifth generations were used. HMVECs were divided into normal group, thrombin group (T group), natrual hirudin group (H group), thrombin + natrual hirudin group (T + H group), AG490 group, thrombin + AG490 group (T + AG490 group), natrual hirudin + AG490 group (H + AG490 group), thrombin + natural hirudin + AG490 (T + H + AG490 group).Apart from the normal group, the other groups were exposed to the relevant drugs for 24 hours.HMVEC apoptosis was assessed by flow cytometric and double Immunofluorescence of phosphorylation of JAK (P-JAK2) and TUNEL assay. Results: Compared with the normal group, in thrombin group the HMVECs apoptosis rate were significantly increased (P<0.05).The results indicated that the index of apoptosis and the apoptosis rate were improved in cultures treated by natural hirudin (T + H group), relative to cultures with thrombin only (T group). We found that the index of apoptosis and the apoptosis rate in the AG490 + thrombin group were higher than that in the hirudin + thrombin group (P<0.05). Double Immunofluorescence of p-JAK2 and TUNEL assays showed that cells were double positive for P-JAK2 uptake and TUNEL detection liquid binding. Conclusion: The natural hirudin and JAK2/STATs signal inhibitor AG490 could block the effects of thrombin. Natural hirudin could attenuate HMVECs apoptosis via antagonizing thrombin and it is suggested that this effect may occur by blocking the JAK2/STATs signaling pathway and this signaling pathways appears to be not the only pathway.
Subject(s)
Humans , Thrombin/drug effects , Antithrombins/pharmacology , Hirudins/pharmacology , Apoptosis/drug effects , Endothelial Cells/drug effects , Protein Kinase Inhibitors/pharmacology , Signal Transduction/drug effects , Cell Proliferation/drug effects , Microvessels/drug effects , Microvessels/metabolism摘要
OBJECTIVE@#To investigate the antithrombotic effects of recombinant hirudin and its mechanism.@*METHODS@#Sixty male Kunming mice were randomly divided into 6 group (=10):control group, model group, aspirin (25 mg/kg) group, recombinant hirudinlow, middle and high dose (0.05, 0.1, 0.2 mg/kg) groups.Except mice in control group, 2.5 mg/kg carrageenan was injected intraperitoneallyto mice in the other groups to produce thrombosis on the mice tail. The mice in aspirin group were administrated intraperitoneally 25 mg/kg aspirin, the mice in recombinant hirudinlow, middle and high dose groups were administrated intraperitoneally 0.05, 0.1, 0.2 mg/kg combinanthirudin, the mice in control group and model group were administrated intraperitoneallynormal saline at the same volume respectively at 24 h, 0.5 h before injecting carrageenan and 24 h after injecting carrageenan. The black tail length of mice and the incidence of black tail were observed at 48h after injection of carrageenan; prothrombin time (PT), activated partial thromboplastin time (APTT), tissue plasminogen activator (t-PA), type-1 plasminogen activator inhibitor (PAI-1), 6-keto-PGF1α, and thromboxane B2 (TXB2) level in mice plasma were determined.@*RESULTS@#As compared with control group, the mice in model group presented tail thrombosis; PT level in plasma was significantly shortened (<0.01), PAI-1 and TXB2levels in plasma were significantly increased (<0.01), while the t-PA and 6-keto-PGF1α levels in plasma in model group were significantly decreased (<0.01). As compared with model group, the thrombus length in the tail was significantly shortened (<0.05, <0.01), PT level was obviously prolonged (<0.01), and the plasma levels of PAI-1 and TXB2 were significantly decreased (<0.01), while the plasma levels of t-PA and 6-keto-PGF1α were significantly increased (<0.01)in the mice of recombinant hirudin low dose, middle dose, high dose groups and aspirin group. As compared with aspirin group, the thrombus length in the tail was significantly increased (<0.05), PT level was obviously shortened (<0.01), and the plasma levels of PAI-1 and TXB2 were significantly increased (<0.01)in the mice of recombinant hirudin low dose group; the plasma level of 6-keto-PGF1α was significantly decreased (<0.01, <0.05) in the mice of recombinant hirudin low dose and middle dose groups; the plasma levels of PAI-1 and TXB2 were significantly increased (<0.01, <0.05)in the mice of recombinant hirudin middle dose group.@*CONCLUSIONS@#The recombinant hirudin can fight against thrombosis, its antithrombotic mechanisms may be related to its influence on the exogenous coagulation system and the promotion of fibrinolysis function.
Subject(s)
Animals , Male , Mice , Blood Coagulation , Fibrinolytic Agents , Hirudins , Pharmacology , Recombinant Proteins , Thromboxane B2 , Tissue Plasminogen Activator摘要
Background: Extracorporeal membrane oxygenation (ECMO) is used with increasing frequency in patients with respiratory and cardiac failure. The achievement of an adequate anticoagulation is critical to avoid patient and circuit complications. Aim: To assess the feasibility and safety of anticoagulation with bivalirudin, as an alternative to unfractionated heparin (UFH), in patient with ECMO. Material and Methods: Observational study, which included all patients receiving anticoagulation with bivalirudin during ECMO, according to a standardized protocol, between august 2015 to May 2016. Results: Bivalirudin was used in 13 out 70 patients connected to ECMO. Ten procedures were for cardiac support and three for respiratory support. Mortality was 43%. ECMO lasted 31 ± 31 days. The time of UFH use before changing to bivalirudin was 7 ± 7 days. The reasons to change to bivalirudin were inadequate levels of partial thromboplastin time (PTT) in nine patients, and heparin induced thrombocytopenia (HIT) in four patients. The time of bivalirudin use was 24 ± 33 days. Per patient, a mean of 2.7 ± 4 oxygenators were changed. These had a useful life of 11.4 and 19.1 days during UFH and bivalirudin use, respectively. The mean bivalirudin dose was 0.08 ± 0.04 mg/kg/h. There was no significant bleeding, thrombosis or circuit obstruction during its use. PTT levels (p < 0.01) and platelet count (p < 0.01) increased significantly after the start of bivalirudin use in patients with UHF resistance and HIT, respectively. Conclusions: Bivalirudin was a safe and efficient drug for anticoagulation during ECMO. It is important to have an alternative drug for anticoagulation in ECMO patients.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Young Adult , Peptide Fragments/blood , Extracorporeal Membrane Oxygenation , Hirudins/blood , Anticoagulants/blood , Partial Thromboplastin Time , Peptide Fragments/administration & dosage , Platelet Count , Recombinant Proteins/administration & dosage , Recombinant Proteins/blood , Heparin/adverse effects , Feasibility Studies , Hirudins/administration & dosage , Anticoagulants/administration & dosage摘要
<p><b>OBJECTIVE</b>This study aimed to investigate the effects of hirudin on the expression of transforming growth factor (TGF-β1) and basic fibroblast growth factor (bFGF) in human gingival fibroblasts (HGFs) in vitro, as well to explore its func- tion in the mechanism of gingival remodeling.</p><p><b>METHODS</b>After culturing was performed with classic tissue-explant method, HGFs were derived from normal gingival and gingival hyperplasia tissues followed by orthodontic treatments with different concentrations of hirudin. The mRNA and protein expression levels of TGF-β1 and bFGF were respectively detected by real time quantity polymerase chain reaction and immunocytochemistry.</p><p><b>RESULTS</b>Compared with normal HGFs, TGF-β1 expression promoted collagen synthesis of fibroblasts, whereas bFGF collagen synthesis was decreased in hyperplasia HGFs without hirudin (P < 0.05). Hirudin significantly upregulated the expression levels of bFGF but downregulated TGF-β1 in hyperplasia HGFs (P < 0.05).</p><p><b>CONCLUSION</b>Orthodontic force may influence the balance of collagen synthesis and degradation in HGFs. Hirudin may modulate the balance of HGF collagen metabolism, thereby promoting gingival remodeling.</p>
Subject(s)
Humans , Fibroblast Growth Factor 2 , Fibroblasts , Gingiva , Hirudins , RNA, Messenger , Transforming Growth Factor beta , Transforming Growth Factor beta1摘要
<p><b>OBJECTIVE</b>To explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.</p><p><b>METHODS</b>Totally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).</p><p><b>CONCLUSION</b>Hirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.</p>
Subject(s)
Animals , Mice , Aorta , Apolipoproteins E , Metabolism , Atherosclerosis , C-Reactive Protein , Cholesterol , Diet, High-Fat , Drugs, Chinese Herbal , E-Selectin , Hirudins , Metabolism , Interleukin-6 , Lipids , Lipoproteins, HDL , Lipoproteins, LDL , Mice, Inbred C57BL , Mice, Knockout , Plaque, Atherosclerotic , Metabolism , Recombinant Proteins , Metabolism , Triglycerides摘要
<p><b>BACKGROUND</b>Bivalirudin was widely used as an anticoagulant during coronary interventional procedure in western countries. However, it was not available in China before this clinical trial was designed. This randomized, single-blind and multicenter clinical trial aimed to evaluate the efficacy and the safety of domestic bivalirudin during percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>A randomized, single-blind, multicenter trial was designed. Elective PCI candidates in five centers were randomized into a bivalirudin group and a heparin group, which were treated with domestic bivalirudin and non-fractional heparin during the PCI procedure. The efficacy was evaluated by comparing the activated coagulation time (ACT), the procedural success rate (residual stenosis < 20% in target lesions without any coronary artery related adverse events within 24 hours after PCI), and the survival rate without major adverse cardiac events at 30 days after PCI between the two groups. Safety was evaluated by the major/minor bleeding rate.</p><p><b>RESULTS</b>A total of 218 elective PCI patients were randomized into a bivalirudin group (n = 110) and heparin group (n = 108). Except for two patients needing additional dosing in the heparin group, the ACT values of all other patients in both groups were longer than 225 seconds at 5 minutes after the first intravenous bolus. Procedural success rates were respectively 100.0% and 98.2% in the bivalirudin group and heparin group (P > 0.05). Survival rates without major adverse cardiac events at 30 days after PCI were 100.0% in the bivalirudin group and 98.2% in the heparin group (P > 0.05). Mild bleeding rates were 0.9% and 6.9% (P < 0.05) at 24 hours, and 1.9% and 8.8% (P < 0.05) at 30 days after PCI in the bivalirudin group and heparin group respectively. There was one severe gastrointestinal bleeding case in the heparin group.</p><p><b>CONCLUSIONS</b>Domestic bivalirudin is an effective and safe anticoagulant during elective PCI procedures. The efficacy is not inferior to heparin, but the safety is superior to heparin.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Antithrombins , Therapeutic Uses , Heparin , Therapeutic Uses , Hirudins , Peptide Fragments , Therapeutic Uses , Percutaneous Coronary Intervention , Recombinant Proteins , Therapeutic Uses , Single-Blind Method , Survival Rate , Whole Blood Coagulation Time摘要
BACKGROUND: The Multiplate analyzer (Dynabyte GmbH) has been recently introduced as a platelet function test for patients taking antiplatelet drugs. The study aimed at providing basic data for determining the reference interval of parameters produced by Multiplate in Koreans and to study the factors that influence those parameters. METHODS: Blood was collected from 35 healthy volunteers (female 18, male 17) into tubes containing hirudin or 3.2% sodium citrate. Whole blood platelet aggregations triggered by adenosine-5'-diphosphate (ADP), ADP-high sensitive (ADP+PGE1 only in hirudin samples), arachidonic acid (AA), collagen or thrombin receptor activator peptide (TRAP) were investigated using Multiplate according to the manufacturer's instructions. Data from healthy volunteers for the area under the curve (AUC) were determined from the central 95th percentile of the results. RESULTS: The values of AUC in hirudin samples for all agonists were significantly higher than those in sodium citrate samples. The AUC values in hirudin (sodium citrate) samples were as follows: ADP 38-107 (18-119) U; ADP+PGE1 16-91 U; AA 64-156 (32-117) U; collagen 53-112 (26-108) U; and TRAP 81-163 (49-149) U. The parameters from Multiplate were significantly correlated with leukocyte counts, but not with hematocrit levels. CONCLUSIONS: Although our data were derived from only 35 subjects, the results are expected to be helpful in determining the reference interval at a single institute and may serve as basic data for future cumulative data of reference intervals from multiple institutes in Korea.
Subject(s)
Humans , Male , Academies and Institutes , Adenosine Diphosphate , Arachidonic Acid , Area Under Curve , Blood Platelets , Citrates , Citric Acid , Collagen , Hematocrit , Hirudins , Korea , Leukocyte Count , Platelet Aggregation Inhibitors , Platelet Function Tests , Receptors, Thrombin , Sodium摘要
OBJECTIVE: Over-expression of thrombin in ovarian cancer cells is associated with poor prognosis. In this study, we investigated the role of thrombin in inducing epithelial-mesenchymal transition (EMT) in SKOV3 epithelial ovarian cancer cells. METHODS: After thrombin treatment SKOV3 cells were subjected to western blots, reverse-transcription PCR, and enzyme-linked immunosorbent assay to quantify EMT-related proteins, mRNA expression of SMAD2, DKK1, and sFRP1, and the secretion of matrix metalloproteinases (MMPs) and cytokines. Meanwhile, invasion ability was evaluated using transwell assays. RESULTS: The results indicated a dose- and time-dependent down-regulation of E-cadherin and upregulation of N-cadherin and vimentin in thrombin-treated SKOV3 cells, compared with the thrombin-free control group (p<0.05). There was a dose- and time-dependent increase in the levels of SMAD2 and DKK1 mRNAs and a decrease in the levels of sFRP1 mRNA in thrombin-treated SKOV3 cells compared to control cells (p<0.05). Thrombin-treated SKOV3 cells exhibited increased secretion of MMP-9, MMP-2, interleukin (IL)-8, and IL-6 and increased invasion compared to untreated cells (p<0.05). Thrombin altered the morphology of SKOV3 cells to a spindle-like phenotype. Addition of hirudin to thrombin-treated cells reversed the effects of thrombin. CONCLUSION: Thrombin induced EMT and promoted the invasion of SKOV3 cells, possibly via distinct signaling pathways. Hirudin inhibited the effects of thrombin, suggesting that anticoagulant therapy could be a novel therapeutic strategy for ovarian carcinoma.
Subject(s)
Blotting, Western , Cadherins , Cytokines , Down-Regulation , Enzyme-Linked Immunosorbent Assay , Epithelial-Mesenchymal Transition , Hirudins , Interleukin-6 , Interleukins , Matrix Metalloproteinases , Neoplasms, Glandular and Epithelial , Ovarian Neoplasms , Phenotype , Polymerase Chain Reaction , Prognosis , Proteins , RNA, Messenger , Thrombin , Up-Regulation , VimentinSubject(s)
Antibodies/analysis , Anticoagulants/adverse effects , Antithrombins/therapeutic use , Benzimidazoles/therapeutic use , Chondroitin Sulfates/therapeutic use , Dermatan Sulfate/therapeutic use , Fibrinolytic Agents/therapeutic use , Heparin/adverse effects , Heparin/immunology , Heparitin Sulfate/therapeutic use , Hirudins , Peptide Fragments/therapeutic use , Pipecolic Acids/therapeutic use , Platelet Aggregation Inhibitors/therapeutic use , Polysaccharides/therapeutic use , Pyridines/therapeutic use , Recombinant Proteins/therapeutic use , Thrombin/antagonists & inhibitors , Thrombocytopenia/chemically induced , Thrombocytopenia/therapy摘要
<p><b>OBJECTIVE</b>To study the extraction system of hirudin emulsion liquid membrane with the Poecilobdella manillensis as raw material, di-(2-ethylhexyl) phosphate (D2EHPA) as carrier, Span 80 as emulsifier, octane and D2EHPA mixed to constitute membrane solution, diluted HCl solutions as internal aqueous phase.</p><p><b>METHOD</b>Using the orthogonal experiment to optimize the extraction conditions of hirudin reference substance such as membrane phase, internal aqueous phase volume ratio (MIPVR), external aqueous phase pH, internal aqueous phase pH, mobile carrier concentration and so on, and then using hirudin crude extracts to do purifying experiment, and gaining experimental samples.</p><p><b>RESULT</b>The optimal conditions of hirudin extraction were as follows: MIPVR 10: 3, internal aqueous phase pH 2.6, external aqueous phase pH 3.4, the mass fraction of carrier D2EHPA 2%. In the optimal extraction conditions, when the initial concentration of hirudin was one anti-thrombin activity units (ATU) x mL(-1), ATU recovery rate of the reference substance was 83.06%. In the purifying experiment of crude extracts, ATU recovery rate was 82.99%, and the specific activity of sample was 3 289.48 the ATU x mg(-1). Discontinuous polyacrylamide gel electrophoresis and spectral scanning, the results showed that the purity and reference substance were considerable.</p><p><b>CONCLUSION</b>The method of preparation hirudin was relatively simple, the purity of the experimental samples and ATU recovery were both high.</p>
Subject(s)
Animals , Emulsions , Chemistry , Hirudins , Leeches , Chemistry , Membranes, Artificial , Solid Phase Extraction , Methods摘要
<p><b>OBJECTIVE</b>To observe the effect of local injection of recombinant hirudin on survival of skin flaps with venous congestion in a rabbit model.</p><p><b>METHODS</b>Eighteen healthy rabbits were enrolled and divided into heparin-treatment (HT), recombinant hirudin treatment (RHT) and control (C) groups according to the random number table, with 6 rabbits in each group. After intravenous anesthesia with 20 g/L pentobarbital sodium, model of skin flaps with venous congestion in the size of 6 cm × 3 cm was reproduced in the dorsal side of left ear of each rabbit, in which central artery of ear served as the only blood supply, and a pedicle of 1 cm in width including central vessel of ear and its accompanying nerves as the only venous return pathway. Each flap in RHT, HT, C groups was respectively given 1 mL recombinant hirudin (1 U), low-molecular-weight heparin (625 U), and isotonic saline via multi-point and homogenous injection, then they were sutured in site. Appearance and survival rate of the flaps were observed after operation. Specimens of the distal part of flaps were harvested for determination of thromboxane B2 (TXB2) on post operation day (POD) 1, 3, 5, 7. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>Rabbit model of skin flaps with venous congestion was reproduced successfully. Obvious hair loss was observed in completely necrotic parts of flap in each group. Obvious edema was observed in all flaps with venous congestion at distal site. The color of flaps in HT and RHT groups were lighter as compared with that in C group, and apparent hematoma of flap was observed in 1 rabbit of RHT group, 2 rabbits of HT group, 4 rabbits of C group on POD 1. The survival rate of flap in HT and RHT groups was respectively (92.3 ± 1.7)% and (94.8 ± 1.9)%, both higher than that in C group [(77.9 ± 1.2)%, F = 191.29, P < 0.05]. There was no statistical difference in survival rate of flap between HT group and RHT group (t = 2.75,P > 0.05). The content of TXB2 in HT and RHT groups on POD 3, 5 was respectively lower than that in C group (with t value from 6.68 to 30.55, P values all below 0.01), but there was no statistical difference between HT and RHT groups (with t value respectively 1.22, 6.44, P values all above 0.05).</p><p><b>CONCLUSIONS</b>Local injection of low-molecular-weight heparin or recombinant hirudin can significantly ameliorate venous congestion of skin flap in rabbit ear, and improve its survival rate.</p>
Subject(s)
Animals , Rabbits , Ear , Graft Survival , Hirudins , Pharmacology , Hyperemia , Recombinant Proteins , Pharmacology , Skin , Surgical Flaps摘要
<p><b>OBJECTIVE</b>To investigate the effect of Hirudin on random skin flap survival in rats.</p><p><b>METHODS</b>24 SD rats were randomly divided into control group and experimental group. The "McFarlane flap (3 cm x 9 cm)" rat models were established on the rat dorsum. 3 ml Hirudin (30 ATU) was injected into the flap in the experimental group, while 3 ml saline in the control group. The injection was performed for 7 days. The flap survival area in the two groups was measured. The tissue samples were taken from proximal (I), middle (II) and distal (III) portions of flaps for histologic study. The VEGF and bFGF expression was also detected with immunohistochemistry method.</p><p><b>RESULTS</b>7 days after operation, the flap survival rate was (69.52 +/- 3.23)% in the experimental group, while (50.36 +/- 2.37)% in control group, showing a significant difference between the two groups (P < 0. 01). In the middle portion, tissue edema and infiltration of neutrophils in experimental group was markedly slighter than that in control group. The VEGF and bFGF expression and neovascularization was enhanced markedly in experimental group.</p><p><b>CONCLUSIONS</b>Hirudin can increase the survival of random pattern skin flaps. It may increase the VEGF, bFGF expression through a series of complex regulatory pathway. Then flap neovascularization is promoted and the flap blood supply is increased.</p>
Subject(s)
Animals , Male , Rats , Fibroblast Growth Factor 2 , Metabolism , Graft Survival , Hirudins , Pharmacology , Rats, Sprague-Dawley , Skin Transplantation , Surgical Flaps , Vascular Endothelial Growth Factor A , Metabolism摘要
Heparin is one of the most frequently used anticoagulants. It is easy to use, but can be associated with life-threatening side effects. One of these is heparin-induced thrombocytopenia syndrome [HITS], which develops in about 3-5% of patients exposed to heparin and is associated with thrombosis in 1% of cases. We report here the successful treatment of five patients with HITS who were treated with alternative anticoagulants namely danaparoid or hirudin. The median time between their exposure to heparin and onset of symptoms and or signs was 10.2 days [range 7-14 days]. Platelet counts decreased to a mean of 38.4 x 10[9] /I [12-82 x 10[9]/1]. All five patients had evidence of thrombosis; four patients had clinical and radiological evidence of pulmonary emboli, one patient had confirmed deep vein thrombosis [DVT] and one patient had extensive skin necrosis of the thighs and abdomen. Platelet aggregation test were positive in two patients, inconclusive in one patient and negative in two patients. Two patients were anticoagulated with danaparoid and three with hirudin until their platelet counts returned to normal between 4 and 14 days [average 6 days] following the recognition of the syndrome. Our patients had significant morbidity, but no mortality. Immediate withdrawal of heparin is of paramount importance and introduction of alternative anticoagulant is necessary in the presence of thrombosis
Subject(s)
Humans , Male , Female , Middle Aged , Aged , Adult , Heparin/adverse effects , Anticoagulants/adverse effects , Anticoagulants , Hirudins , Platelet Count , Dermatan Sulfate , Pulmonary Embolism , Venous Thrombosis摘要
To establish a refolding process for the protein fused with 12-peptide of hirudin and reteplase (HV12p-rPA), we developed an anion-exchange chromatography assisted method to form correct disulfide bonds. After evaluating various parameters by orthogonal experiments with Q Sepharose XL as refolding medium, we found that urea gradient, sample loading size and L-Arg concentration were three major factors to affect the refolding outcomes, and urea gradient was critical to the recovery yield. Meanwhile, enzymatic activity of the refolded protein was decreased by the increase of sample loading size, and the optimal concentration of L-Arg in the eluting buffer was 1 mol/L. Thus, a dual-gradient of urea and pH on the anion-exchange chromatography resulted in remarkable increase of specific fibrinolytic and anti-coagulative activities of the refolded protein. Compared with the dilution method for refolding HV12p-rPA, the present approach was more effective and advantageous.
Subject(s)
Chromatography, Ion Exchange , Methods , Disulfides , Chemistry , Fibrinolytic Agents , Chemistry , Hirudins , Chemistry , Protein Refolding , Recombinant Fusion Proteins , Chemistry , Recombinant Proteins , Chemistry , Tissue Plasminogen Activator , Chemistry摘要
As a potent anticoagulant, leech a traditional Chinese medicine, has become increasing topics. Hirudin, which is the primary effective component in leech, is a specific and efficient inhibitor of thrombin, mainly used in prevention and treatment of thrombus on the clinic practice. However, there is still no accurate and convenient method reported about the determination of it's biological activity. This paper reported a method for the determination of the biological activity the of extract from hirudo. The extra thrombin, which was not inhibited by hirudin in the extract from hirudo, reacted with N-benzoyl-L-arginine ethyl ester and was determined. The biological activity of the hirudo extract was determined, indirectly. The linear of calibration curve and accuracy were both perfect, the method was accurate and reliable.
Subject(s)
Animals , Arginine , Biological Factors , Pharmacology , Enzyme Assays , Methods , Enzyme Inhibitors , Pharmacology , Hirudins , Pharmacology , Hirudo medicinalis , Chemistry , Thrombin摘要
<p><b>OBJECTIVE</b>To study the effect of hirudin on the function of human hyperplastic scar fibroblasts (HSFBs).</p><p><b>METHODS</b>HSFBs were cultured in vitro. Hirudin solution in the concentration of 1, 10, and 50 kU/L was respectively added into DMEM culture medium to form 1, 10, and 50 kU/L hirudin groups, with 9 wells in each group. HSFBs cultured without hirudin were set up as control group. Cell inhibition rate, secretion level of TGF-beta1 from cells, and expression levels of mRNA of type I and III precollagen were determined at 24, 48, and 72 h after culture.</p><p><b>RESULTS</b>Inhibition rates of HSFBs growth was respectively (29.3 +/- 0.9)%, (30.1 +/- 0.3)%, and (45.2 +/- 1.9)% when cultured with 10 kU/L hirudin for 24, 48, and 72 hs, which were higher than those in control group [(0.0 +/- 0.0)%, P < 0.05]. There was statistically significant difference between control group and 1 and 50 kU/L hirudin groups in the inhibition rates of HSFBs at some time points (P < 0.05). Secretion level of TGF-beta1 of HSFBs in 1, 10, 50 kU/L hirudin groups was respectively (228.5 +/- 1.8), (210.5 +/- 11.1), and (168.5 +/- 14.1) pg/mL when cultured for 48 hs, of which the last 2 figures were significantly lower than that of control group [(265.0 +/- 1.5) pg/mL, P < 0.05]. Hirudin in the concentration of 10 and 50 kU/L could inhibit the expression of mRNA of type I and III precollagen in HSFBs.</p><p><b>CONCLUSIONS</b>Hirudin solution in the concentration of 10 and 50 kU/L can inhibit the proliferation of HSFBs and secretion of TGF-beta1 and collagen in certain degree.</p>
Subject(s)
Humans , Cells, Cultured , Cicatrix, Hypertrophic , Pathology , Fibroblasts , Cell Biology , Bodily Secretions , Hirudins , Pharmacology , Transforming Growth Factor beta1 , Metabolism摘要
BACKGROUND: Current treatment strategies for percutaneous coronary intervention (PCI) and acute coronary syndrome (ACS) include concomitant use of glycoprotein IIb/IIIa inhibitors (GPI) and antithrombotic therapy such as aspirin, clopidogrel, and unfractionated or low-molecular-weight heparin. The "direct thrombin inhibitor" bivalirudin has been associated with better efficacy and safety than heparin. OBJECTIVE: The present study is performed to evaluate the safety and efficacy of an indigenously developed and manufactured bivalirudin (Bivaflo; Sun Pharmaceutical Industries Ltd., Mumbai) as the primary anticoagulation strategy during PCI in moderate-high risk patients with only provisional use of GPI. METHODS: This prospective multicentered registry enrolled 439 patients in 11 tertiary care centers across India. Patients who had ACS or other clinical/angiographic characteristics, which increase risk during PCI, were enrolled in the registry. Bivaflo was administered as a bolus dose of 0.75 mg/kg, followed by infusion at a rate of 1.75 mg/kg/h during the procedure and optionally 0.25 mg/kg/h for 4 hours after the procedure at investigator's discretion. GPI use was discouraged except as bailout. The primary endpoints were composite and individual incidences of death, myocardial infarction (MI), urgent revascularization, subacute stent thrombosis (SAT), or bleeding at day 7/hospital discharge, whichever was earlier. The secondary endpoints were 30-day composite and individual incidences of death, MI, urgent revascularization, and SAT. RESULTS: The mean age of the group was 58 +/- 10 years and 83% were males. Bivaflo was administered for a mean duration of 102 +/- 79 minutes, and 65% patients received Bivaflo infusion post-PCI. ACT values measured at 10 minutes after bolus and at the end of the PCI were found to be 339 +/- 110 and 336 +/- 104 seconds, respectively. GPI was provisionally used in only 4% (16) patients mostly due to new or suspected thrombus and obstructive dissection with decreased flow. At day 7/hospital discharge, there were no incidences of major adverse cardiac events or major bleeding. Minor bleeding occurred in only 4 patients (0.9%). The 30-day composite major adverse cardiac event rate was 0.68%. One death and two subacute thrombosis occurred during the 30-day follow-up. CONCLUSION: Bivaflo is safe and effective sole anticoagulation strategy during PCI of moderate-high risk patients. Bivaflo administration was associated with no major bleeding events and extremely low in hospital and 30-day MACE rate. These rates were lower than expected MACE rates for such a subgroup of patients based on historical controls.