Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients. Methods Twenty ASA Ⅰ or Ⅱ patients, aged 20-44 yr, weighing 40-75 kg, required blood salvage during operations (intraoperative blood loss expected 15-20 ml/kg), were randomly divided into 2 groups ( n = 10 each): control group (group C) and leukocyte depletion group (group D) . Anesthesia was induced with midazolam, fentanyl, propofol and vecuronium and maintained with isoflurane inhalation and iv infusion of propofol and remifentanil. The patients were mechanically ventilated after tracheal intubation. Intraoperative blood salvage and reinfusion were performed in all patients using cell saver system. In group D, the salvaged blood was filtered with a leukocyte depleting filter placed in the line of the reinfusion circuit. Blood samples were collected from the central vein before anesthesia (T1 ) , at the end of surgery (T2 ) , and at 12 h (T3 ) and 36 h (T4 ) after operation in the two groups. The rosette rates of RBC-C3b receptors and RBC-immune complex were determined. The leukocyte and neutrophil were counted. The plasma levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured. Results The plasma levels of IL-6, TNF-a and MDA were significantly lower at T2-4 , the rosette rate of RBC-C3b receptors was significantly higher at T3-4 ( P ＜ 0.01) , and the neutrophil count was significantly lower at T4 in group D than in group C ( P ＜ 0.05) . Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients, and the decrease in the systemic inflammatory response may be involved in the mechanism.

Objective: To study the protective effect of coenzyme Q_10 on red blood cell membrane and its immunity in patient during extracorporeal circulation. Method: Twenty patients under extracorporeal circulation about 30 minutes were randomly divided into control group and coenzyme Q_10 group (n=10). Coenzyme Q_10 2mg/kg was added to the priming fluid in coenzyme Q_10 group. Plasma levels of malondialdehyde (MDA), free hemoglobin (FHb), immune adhe sion ability and immune compound of red blood cell membrane were measured before extracorporeal circulation, 15 and 30 minutes after beginning of cardiopulmonary bypass, and on first day morning after operation. Result; All different periods after beginning of cardiopulmonary bypass in coenzyme Q_10, group, MDA and FHb levels were lower and im mune adhesion ability was higher than those of control group. Conclusion: Coenzyme Q_10 may protect red blood cell membrane and its immunity of patient during extracorporeal circulation.

Objective To investigate the effect of leukocyte-depleted intraoperative salvaged blood on the erythrocyte immunity and systemic inflammatory response during the perioperative period in patients.Methods Twenty ASA Ⅰ or Ⅱ patients (aged 20-44 years and weighing 40-75 kg) requiring blood salvage during operation (with the estimated intraoperative blood loss of 15-20 ml/kg) were randomly divided into two groups (n =10 each):control group (group C) and leukocyte depletion group (group D).Anesthesia was induced with midazolam,fentanyl,propofol and vecuronium and maintained with isoflurane inhalation and intravenous infusion of propofol and remifentanil.The patients were mechanically ventilated after endotracheal intubation.Intraoperative blood salvage and reinfusion were performed in all the patients.In group D,the salvaged blood was filtered by a leukocyte depletion filter placed in the line of the reinfusion circuit.Blood samples were collected from the central vein before anesthesia (T1),at the end of surgery (T2) and 12 hours (T3) and 36 hours (T4) after operation in all the patients.The rosette rates of red blood cell-C3b receptors (RBC-C3bRR) and RBC-immune complex (RBC-ICR) were determined.The number of leukocytes and polymorphonuclear neutrophils (PMNs) were counted.The plasma levels of interieukin-6 (IL-6),tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA) were measured,too.Results Compared with group C,the plasma levels of IL-6,TNF-α and MDA were significantly lower at T2-T4,the RBC-C3bRR was significantly higher at T3-T4 (P ＜ 0.01),and the number of PMNs was significantly lower at T4 in group D(P＜0.05).Conclusion Leukocyte-depleted intraoperative salvaged blood is helpful to improve the erythrocyte immunity during the perioperative period in patients,and the decrease of systemic inflammatory response may be involved in the mechanism.

Objective To evaluate the effects of pyruvate Ringer' s solution on erythrocyte deformation function in the patients undergoing cardiopulmonary bypass (CPB) in vitro.Methods Thirteen patients of both sexes,aged 25-64 yr,of ASA physical status Ⅱ or Ⅲ (NAHY Ⅱ or Ⅲ),scheduled for elective cardiac valve replacement under CPB were enrolled in the study.At 10 and 60 min of CPB (T1,2) and 60 min after termination of CPB (T3),blood samples 18 ml were drawn from the radial artery and each patient's blood sample was equally divided into 6 parts (3 ml per part).The blood samples were divided into 3 groups (n =13 each):control group (group C),lactated Ringer's solution group (group R),and pyruvate Ringer's solution group (group P).In C,R and P groups,normal saline 1 ml,lactated Ringer's solution 1 ml,and pyruvate Ringer's solution 1 ml were added,respectively,and then the erythrocytes were incubated for 60 min at 37 ℃.At 30 and 60 min of incubation,the erythrocyte rigidity index (ERI) and erythrocyte deformation index (EDI) were detected.Results Compared with C and R groups,the ERI was significantly decreased at 30 and 60 min of incubation at T1,the EDI was decreased at 30 min of incubation at T1,2,and no significant change was found in ERI at T2,3 or in EDI at T3 in group P.There was no significant difference in ERI and EDI at different time points of incubation within groups.Conclusion Pyruvate Ringer's solution can enhance erythrocyte deformation function in the patients undergoing CPB.

ObjectiveTo investgate the effects of pyruvate sodium on energy metabolism of erythrocytes in dog blood during circulation in cardiopulmonary bypass machine.MethodsTen healthy adult mongrel dogs of both sexes weighing 12-18 kg were anesthetized with intraperitoneal 2.5％ pentobarbital 25 mg/kg.Femoral artery was cannulated and 500 ml of blood was collected from femoral artery in each dog.Blood samples from the 10 dogs were divided into 2 equal parts (250 ml each).Each part was mixed with 250 ml lactated Ringer's solution (group C) or 250 ml pyruvate sodium (group P).Flow rate of 4 L/min was employed and blood temperature was kept at 30-32 ℃ during circulation in cardiopulmonary bypass machine.The blood sanples were collected before (baseline value,T1 ) and at 30,60,90,120 min of circulation (T2-5) for determination of ATP,ADP and AMP concentrations in RBC.ResultsThe ATP and AMP concentrations were significantly higher at T2-5,while ADP concentration was higher in group P than in group C.ConclusionPyruvate sodium can improve energy metabolism of erythrocytes in dog blood druing circulation in cardiopulmary bypass machine.

Objective To explore and evaluate the changes of hemorheology and fracture healing in patients undergoing spinal operation with intraoperative autologuous blood transfusion.Methods A total of 80 patients underwent internal fixation of spine in our hospital from January 2012 to December 2014 were selected in this study.Digital random table method were utilized for dividing them into two groups.Autologous blood transfusion was used in group A,allogeneic blood transfusion was applied in group B.Blood viscosity,plasma viscosity,hematocrit,fibrinogen,erythrocyte sedimentation rate (ESR) and the level of sICAM-1,sVCAM-1 were collected and compared 1,4,8,12 weeks after the operation.Fesults Concentrated red cell and plasma volume infusion in group A was lower than that in group B,the level of sICAM-1,sVCAM-1 1,4,8,12 weeks after operation in group A were significantly lower than that in group B.There was no delayed fracture healing patients in group A,but there was 3 patients with delayed fracture healing in group B (P＜0.05).Blood viscosity,plasma viscosity,hematocrit,fibrinogen,erythrocyte and ESR 1,4,8,12 weeks after the operation compared in both group were not statistically significant (P＞0.05).Conclusion Autologous blood transfusion is not only a safe and effective method for blood transfusion,but also can improve the tissue blood supply,reduce inflammation,and conducive to fracture healing.

Objective:To evaluate the role of caspase-3 in aggravation of the oxidative stress injury in stored red blood cells (sRBCs) by cardiopulmonary bypass (CPB).Methods:Eight patients with type O blood undergoing valve replacement with CPB were selected, blood samples 7 ml were collected through the central venous catheter before CPB, blood samples 20 ml were collected at 2 h after CPB, and the plasma before and after CPB was obtained after centrifugation. Eight samples of sRBCs 14 ml of blood type O stored for 7-14 days were collected and each sample was divided into A, B, C and D groups with 3.5 ml in each group. Normal saline 30 μl was added to group A and group B, 10% dimethyl sulfoxide 30 μl was added to group C, and 3.5 mmol/L Z-DEVD-fmk solution 30 μl was added to group D. The sRBCs were pretreated in a 37 ℃ water bath for 2 h in the four groups. Then group A was incubated with plasma before CPB, group B, C and D were incubated with plasma at 2 h of CPB, and four groups were incubated for 48 h in a thermostatic oscillator at 37 ℃ and 80 rpm. At 2, 24 and 48 h of incubation, the activity of caspase-3 and concentration of ATP were determined by enzyme-linked immunosorbent assay, the concentrations of glutathione (GSH) and free hemoglobin (FHb) were measured by colorimetry, and the exposure rate of cell membrane phosphatidylserine (PS) was detected by flow cytometry.Results:With the prolongation of incubation time, the activity of caspase-3, exposure rate of PS at cell membrane and concentration of FHb were gradually increased, and the concentrations of ATP and GSH were gradually decreased in the four groups ( P<0.05). Compared with group A, the activity of caspase-3 was significantly increased at each time point of incubation in group B and group C, the activity of caspase-3 was increased at 24 and 48 h of incubation in group D, and the concentration of ATP was decreased at 24 and 48 h of incubation, and the concentration of GSH was decreased and the concentration of FHb was increased at each time point of incubation in group B, group C and group D ( P<0.05). Compared with group B and group C, the activity of caspase-3 was significantly decreased, the concentrations of ATP and GSH were increased, and the exposure rate of PS at cell membrane and concentration of FHb were decreased at 24 and 48 h of incubation in group D ( P<0.05). There was no significant difference in the parameters mentioned above between group B and group C ( P>0.05). Conclusions:Caspase-3 is involved in aggravation of oxidative stress injury in sRBCs by CPB.

Red blood cells (RBCs) are one of the most needed blood products in clinical blood transfusion treatment. At present, RBCs are mainly provided by blood donors, and the contradiction between supply and demand is extremely prominent. Customizable RBCs cultured in vitro bring dawn to solve this contradiction. RBCs can be differentiated and cultured from human induced pluripotent stem cells, immortalized adult erythroid progenitor cells, embryonic stem cells (ESCs), hematopoietic stem cells or the mononuclear cells extracted from umbilical cord blood and peripheral blood. Each method, with its own advantages and drawbacks, are still in the stage of laboratory research. However, according to the current progress, it is not far from clinical application. This paper summarizes the progress in this field.

Objective: To evaluate the effect of compound electrolyte injection on phosphatidylserine (PS) exposure in erythrocytes after blood salvage-retransfusion in dogs. Methods: Twenty healthy mongrel dogs, weighing 10-15 kg, aged 3-5 weeks, were divided into 2 groups (n=10 each) using a random number table method: normal saline group (group NS) and compound electrolyte injection group (group MEI). The process of intraoperative blood salvage-retransfusion was simulated in both groups: femoral vein was cannulated for blood withdrawal until the volume of blood lost was 400 ml, and the shed blood was salvaged by a blood recovery machine.The washing solution was normal saline in group NS and compound electrolyte injection in group MEI.The erythrocytes were retransfused after being labeled with fluorescein isothiocyanate.Blood samples were obtained before and after blood salvage for determination of erythrocyte ATP content by enzyme-linked immunosorbent assay.Blood samples were obtained at 24, 48 and 72 h after blood retransfusion, and the PS exposure rate of the salvaged erythrocytes was determined by flow cytometry.The spleen was taken at 72 h after retransfusion to detect the phagocytosis rate of salvaged erythrocytes by monocytes. Results: There was no significant difference in ATP content before and after blood salvage between the two groups (P>0.05). Compared with NS group, the PS exposure rate of the salvaged erythrocytes at each time point after retransfusion and phagocytosis rate of salvaged erythrocytes by monocytes were significantly decreased in MEI group (P<0.05). Conclusion Compound electrolyte injection as a washing solution for intraoperative blood salvage can reduce the PS exposure in salvaged erythrocytes and is helpful in prolonging the lifespan of erythrocytes after retransfusion in dogs.