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Objective To investigate the preventive effect of low molecular weight heparin or tranexamic acid in the rabbits with hypercoagulation induced by impact injury and intravenous infusion of the endotoxin. Methods Combination of impact injury and endotoxin was created by strike on rabbit chest with type BIM-Ⅱ biological impact machine and injection of endotoxin via vein.Thirty-two male New Zealand rabbits were divided into four groups in a completely random design as follows:impact injury combined with endotoxin without treatment group(control group),treatment with low molecular weight heparin(L group),treatment with tranexamic acid(TA group),and treatment with low molecular weight heparin and tranexamie acid(L+TA group).The changes of interleukin-1β(IL-1β)and interleukin-6(IL-6)were observed by using the enzyme-linked immunosorbent assay;changes of antithrombin-Ⅲ(AT-Ⅲ)activity,tissue factor(TF)and tissue factor pathway inhibitors(TFPI)were detected by the chromogenic substrate method chromozym P:the content of fibrinogen was observed by coagulation analyzer.The lung and kidney were removed from the experimental animals at 24 hours post impact injury forhistopathologic observation. Results In the control group,the activity of AT-Ⅲ was decreased obviously.while the content of fibrinogen and the activity of TF and TFPl were increased,with formation of the thrombosis in the kidney after injection of PLS.Compared with control group,the activity of TF and the content of IL-1β and IL-6 were reduced but the TFPI was increased in the L group,which resulted in a decrease of TF/TFPI ratio.with decrease of formation of the micro-thrombosis in the organs.The content of IL-1β and IL-6 in the TA group were reduced,which was still higher than that in tIle L group.In the TA group.the activity of IF and TFPI Was lower than that in the L group and control group,with higher TF/TFPI ratio than the L group.The frequency of thrombosis increased in pulmonary artery and other important organs.The content of IL-1β and IL-6 in the L+TA group wag lower than that in the L group.In the L+TA group.the activity ofTF was lower than that inthe L group and control group,but the activity of TFPI was higher than that in the L group and control group.with lower TF/TFPI ratio than the L group.There found no increase of thrombus in the pulmonary arterial and other important organs. Conclusions Besides anticoagulation function,the low molecular weight heparin can inhibit the inflanunation reaction,reduce the release of TF and attenuate the coagulation reaction.On the basis of treatment with low molecular weight heparin,application of the tranexamic acid will not lead to the formation of thrombus,but the only use of the tranexamic acid would have the contrary results.The rtsults of the study provide a theoretical and experimental basis for clinical diagnosis and prevention of hypercoagulation induced by injury and infection.
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Objective To study the possible mechanisms of aprotinin in the protection of platelets. Methods Cytosolic free calcium concentration ([Ca 2+ ]i) was determined in calcium fluorescent indicator Fura 2/AM loaded washed human platelets by using dual wavelength spectrofluorophotometer. The mean value of resting [Ca 2+ ]i and the changes of [Ca 2+ ]i response to thrombin and aprotinin were observed. Results In the presence of extracellular Ca 2+ at the dose of 1 mmol/L, the resting level of [Ca 2+ ]i in platelets of human was (151.840?28.719) nmol/L. Thrombin stimulated the rise in [Ca 2+ ]i in the presence of Ca 2+ at the dose of 1 mmol/L, and the effects were inhibited by aprotinin in a concentration dependent manner ( P
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<p><b>OBJECTIVE</b>To study the mechanism of macrophage injury after trauma-hemorrhagic shock.</p><p><b>METHODS</b>Wistar male rats underwent trauma (closed bone fracture) and hemorrhage (mean arterial blood pressure of 35 mm Hg+/-5 mm Hg for 60 minutes, following fluid resuscitation). Rats without trauma, hemorrhage or fluid resuscitation served as controls. Peritoneal macrophages were harvested at 6 hours and 1, 2, 3, 7 days after traumatic hemorrhagic shock to determine the effects of pertussis toxin (PTX, as a specific inhibitor to Gi(alpha) and cholera toxin (CTX, as a stimulant to Gs(alpha) on macrophage-Ia expression and TNF-alpha production and levels of Gi(alpha) and Gs(alpha).</p><p><b>RESULTS</b>The macrophages from the injured rats revealed a significant decrease of Ia positive number and TNF-alpha release in response to LPS. Wi th pretreatment with PTX 10-100 ng/ml Ia positive cells and LPS-induced TNFalpha production in both control and impaired macrophages populations were dos e dependently increased. Both macrophages populations were not responding to CTX treatment (10-100 ng/ml). Western blot analyses showed that the levels of Gi(alpha) protein expression increased as much as 116.5%-148.8% of the control level fro m 6 hours through 7 days after traumatic hemorrhage. The levels of Gs protein expression were reduced at 6 hours and decreased to the lowest degree; 36% o f the control at day 1, began to return at day 2 and returned to the normal level at day 7, following traumatic hemorrhagic shock.</p><p><b>CONCLUSIONS</b>PTX-sensitive G-protein may participate in th e modulation of macrophage-Ia expression and TNF-alpha release following traumatic hemorrhagic shock. Analyses of the alteration of Gi(alpha) and Gs protein express ions further supports the concept that G-protein is involved in trauma-induced macrophage signal transduction pathways.</p>
Subject(s)
Animals , Male , Rats , Analysis of Variance , GTP-Binding Proteins , Allergy and Immunology , Metabolism , Histocompatibility Antigens Class II , Allergy and Immunology , Immunoblotting , Lipopolysaccharides , Pharmacology , Macrophages, Peritoneal , Allergy and Immunology , Metabolism , Rats, Wistar , Shock, Hemorrhagic , Blood , Allergy and Immunology , Signal Transduction , Tumor Necrosis Factor-alphaABSTRACT
Objective To investigate the effect of cardiopulmonary bypass (CPB) on vascular endothelial cell injury and plasma endothelin-1 and nitric oxide equilibrium in patients undergoing cardiovascular operation with CPB. Methods A total of 20 patients with congenital heart disease (Group Ⅰ) and 20 with valvular problem (group Ⅱ) were operated on under CPB respectively. Blood samples were collected from central vein before skin incision, before CPB, 30 min after CPB, at the end of CPB, and end of operation, the first morning and third morning after operation. The levels of plasma thrombomodulin(TM), endothelin-1(ET-1) and nitric oxide(NO) were measured. Results The plasma TM level was significantly elevated during CPB (P<0.01, P<0.05) and 1 d after operation, reached its peak as (4.88±1.12) ng/ml in Group Ⅰand (8.34±1.84) ng/ml in group Ⅱ at the end of surgery and came back to the level as before operation. The plasma level of ET-1 was also increased significantly after CPB and reached peak as (129.04±22.29) in Group Ⅰ and (156.62±29.66) in Group Ⅱ at the end of operation. And the level was still higher than before operation in 2 groups 3 d after operation. No change was found on the level of NO in 2 groups. Conclusion CPB may cause extensive acute endothelial cells damage for about 24-48 h and recovered about 72 h and it may also cause an imbalance of ET-1 and NO.
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Objective To investigate the effects of ketamine on the effector functions (phagocytosis, respiratory burst and release of proteolytic enzymes) of lipopolysaccharide (LPS) stimulated human neutrophils in vitro. Methods Blood samples were collected from healthy volunteers. The study was divided into four groups: LPS group and three ketamine groups (K1, K2 and K3). The final concentration of ketamine in each group was 0, 3, 30 and 300 ?g/ml respectively. Phagocytosis was assessed in whole blood by NBT phagocytosis test (n=8) and respiratory burst by flow cytometry (with dihydrorhodamine 123 as fluorescent marker, n=5). The release of three proteolytic enzymes was measured with isolated polymorphonuclear neutrophils (PMNs) by turbidimetry (lysozyme) and chromatometry (elastase and ? glucurolidase) methods (n=9).Results Ketamine dose dependently inhibited phagocytosis, respiratory burst and proteolytic enzyme releasing of LPS stimulated human neutrophils in vitro. Higher concentrations of ketamine were required to suppress respiratory burst as compared with the concentrations needed to suppress phagocytosis and proteolytic enzyme releasing.Conclusions The inhibitory effects of ketamine on the effector functions of LPS stimulated human neutrophils may contribute to the attenuation of neutrophil mediated inflammatory injuries.
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Objective To investigate the effect of hydroxyl starch (HES 200/0 5) infusion on blood rheology during operation Methods Forty adult ASA Ⅰ Ⅱ patients (male 28, female 12) scheduled for elective operation on spine (26 patients) and lower extremities (14 patients) were enrolled in this study The age ranged from 18 56 yr and weight from 40 85kg Preoperative heart, lung, liver and kidney function tests, blood routine exam, bleeding and coagulation time were all normal No patients had any history of abnormal bleeding The patients were randomly divided into two equal groups: control group(C) receiving lactated Ringer solution 1 000ml during operation and HES group receiving 6% HES 1 000ml No patient received any plasma or whole blood during operation The patients were premedicated with intramuscular atropine 0 5mg and sodium luminal 0 1g The operations were all performed under continuous epidural block (1 6% lidocaine with adrenaline 1:200 000) BP,ECG,and SpO 2 were continuously monitored during operation Blood pressure was maintained with range and SpO 2 above 97% Venuos blood samples were taken before anesthesia , 30min and 4h after the fluid had been infused and 24h after operation for determination of blood viscosity, plasma viscosity, hematocrit(HCT) index of red blood cell (RBC) aggregation and deformation and index of fibrinogen Results Blood viscosity, Hct, index of RBC aggregation and fibrinogen decreased significantly (P
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Objective To investigate the changes in counts of T-lymphocyte subsets, natural killer cell (NK cell) and the levels of serum soluble interleukin-2 receptor(sIL-2R) after perioperative allogeneic whole-blood transfusion or erythrocyte suspention (SAG-M blood) blood transfusion. Methods Thirty patients undergoing elective refection for rectal cancer, were randomly allocated to receiving 400 ml of allogeneic whole-blood (n = 15 ) or 400 ml of SAG-M blood (n = 15 ) during perioperative period, respectively. Indirect immunofluorescence technique and enzyme-linked immunosorbent assay were used to determine the counts of T--lymphocyte subsets, NK cells and to detect levels of serum sIL-2R 1 day and 5 day after operation.Results The counts of CD3 +, CD. + and NK cells in both groups decreased significantly after operation (P
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Objective: To observe the influences of infusing small volume of 7.5% NaCl solution (HS) in vivo,and the different volume ratio of plasm of rich platelets (PRP) to HS or sucrose in vitro on the function of platelet. Method:Twenty-nine patients undergoing middle or small operation were divided randomly into HS group and normal saline group(NS). Blood samples were taken before infusion and,5,30,60 and 120 min after infusion,to measure hematocrit (HCT),platelets (PLT) count, maximum aggression rate (MAR),thromboxance B_2(TXB_2),granule membrane protein-140(GMP-140) and platelet factor 3 activety (PF_(3a)T). In addition,fifteen blood samples of blood donors were taken,and divided control group,HS group,sucrose group and NS group. The volume ratio in each group were arranged in 1/25,2/25,4/25 and 8/25. Osmotic concentration of 1/25 was equivalent to that induced with small volume hypertonic saline infusion. Result: Compare with NS group,the HCT and PLT count were decreased after infusion of small volume of HS. When the volume ratio ≥2/25, all measure values of the MAR, TXB_2, GMP-140 and PF_(3a)T were decreased in vitro,and profoundly inhibited following osmolarity increase of HS. Conclusion;Small volume of HS may be safely used,but severe inhibition of functions of platelet may occur when the infusing volume of HS exceeded more than twice of small volume of HS.
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Obiective: To verify the effects of aminoguanidine on hemodynamics in sepsis.Method:Experiments were conducted in five groups of anesthetized dogs (each group n=8).In group 1,lipopolysaccharide (LPS)was injected intravenously, Group 2 received both LPS and L-arginine (300mg/kg)intravenously. Group 3 received both LPS and N-nitro-Larginine (L-NNA,20mg/kg) intravenously.Group 4 received both LPS and aminoguanidine (30mg/kg) intravenously. Group 5 received only saline. Hemodynamic and oxygenational data were measured or calculated. Extravascular lung water (EVLW)was measured. Result:L-arginine increased CI and decreased PVRI,while treatment with aminoguanidine without significant increased the BP,SVRI and PVRI.But all of untoward hemodynamic effcts of LPS were exacerbated by the addition of L-NNA,as DO_2 was significantly decreased by L-NNA,Therefore,though O_2ER was also increase (insufficiently),VO_2 was still decreased significantly.EVLW was markedly increased by LNNA. Conclusion:The No-selective inhibition with adiministration of aminoguanidine may have considerable value in the therapy of endotoxin shock.
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Objective To investigate the effects of magnitude of surgical trauma and duration of operation on aggregation and adhesion of platelets during laparoscopy and conventional laparotomy cholecystectomy. Methods Thirty ASA Ⅰ-Ⅱpatients(male 11, female 19) undergoing elective cholecystectomy were studied . The mean age was(45 .2 ? 8.3) years and mean body weight(63.3 ?12. 6)kg. The patients were divided into two groups: laparoscopy group(group LC) and laparotomy group(group OC). Patients who took any drugs which may affect blood coagulation were excluded. Blood routine examination, coagulation and bleeding time were normal in all patients. In both groups anesthesia was induced with fentanyl 0. 1-0.2mg, propofol 2mg/kg and vecuronium 0.1mg/kg and maintained with isoflurane inhalation. Venous blood samples were taken after induction of anesthesia and 1h and 2h after operation was started. 10ml of blood was withdrawn from median cubital vein for the measurement of platelet adhesion(PAdT) and plasma level of platelet membrane glycoprotein(GPⅠ b/Ⅰ x) and von Willebrand factor(vWF) . Venepuncture was made at first attempt without using tourniquet. Plastic syringes were used and first 2ml of blood withdrawn was discarded . Glass ball method was used for measurement of PAdT which was calculated according to the following equation: Rate of platelet adhesion(% ) = /(No. of platelet before adhesion-No. of platelet after adhesion) No. of platelet before adhesionGP Ⅰ b/Ⅰ x and vWF levels were measured using ELISA method. Results There was no significant difference in PAdT and GP Ⅰ b/Ⅰ x and vWF levels after induction of anesthesia between two group. PadT level was significantly lower at 1 and 2h during surgery in group LC than that in group OC. GPⅠh/Ⅰx level was higher at 1h during operation in group OC than that in group LC and was much higher at 2h during surgery. vWF level decreased significantly at 2h during operation in both groups. Conclusions Platelet adhesion rate increases during operation and the severer the trauma the higher the platelet adhesion rate. GPⅠ b/Ⅰ x level is higher in group OC during operation than that in group LC probably due to severity of trauma. vWF is consumed during operation.
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Objective To study vascular endothelial cell injury/activation associated with CPB in order to understand the pathophysiology of the complications of patients undergoing cardiovascular operationsMethods Twenty-six patients scheduled for cardiac operations with CPB, were divided into two grourps, group Ⅰ: 14 patients with acyanotic congenital heart diseases undergoing corrective surgical procedures; group Ⅱ: 12 patients undergoing selective cardiac operation for valvular replacement Blood samples were withdrawn from central vein before skin incision,before CPB, 30 min following CPB, at end of CPB, at end of operation, on the first postoperative day and the third postoperative day to measure the levels of circulating endothelial cells(CEC), thrombomodulin(TM) and von Willebrand Factor(vWF).Results The levels of CEC,TM and vWF significantly elevated during CPB and on the first postoperative day in the two groups,as compared with those before operation (P
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The effects of hypertonic saline solution (HS) on prostacyclin (PGI2) and endothelin (ET) were evaluated.30 patients under epidural anesthesia during elective operations were infused with 7.5% HS (n=20) or 5% dextrose (n=10) 4ml/kg before anesthesia.PGI2, ET and hemodynamic indices were measured before and 10, 60 min after infusion.In HS group, the PGI2 increased about 51.4%, ET decreased 49.6% and the aortic compliance increased 37.5%. However.after dextrose infusion, PGI2 increased about 23.7% at 60 min, ET increased but dispersedly, with increased TPR and decreased CO, SV.Systolic pressure were declined over 20% in four out of ten patients.It demonstrated that HS could stimulate the secretion of PGI2 and depress ET. These hormonal changes revealed an important role in the course of decreasing TPR, with improving hemodynamios.
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Rocket immunoelectrophoresis was used to determine the fibrin (ogen) degradation products, the appearance of which is one of the important criteria of the fibrinolytic process. Compared with the plasma protamine paracoagulation test, counterelectrophoresis, or Fi and staphylococcal clumping test, this method was more specific, easier to be reproduced, and more accurate quantitatively. There was no false negative or false positive results. This method was considered to be useful in determining whether there is any increase of the fibrinolytic activity.