| Karl Landsteiner ,an Austrian doctor, discovered ABO blood type in 1900 and then won the Nobel Prize in Medicine and Physiology in 1930.From then on, modern transfusion medicine has established.Now , transfusion has become an effective clinical approach in both peacetime and war.A lot of patients have been rescued through transfusion.But the transfusion of homologous blood poses a series of severe problems. The blood always cannot be transfused in time because the lack of blood,difficulty of storage and transport and the complex tests for compatibility and specifc diseases,especially in war,mishaps and surgery accidents. Patients are much concerned about the safety of transfusion for the immune reaction and the transfution transmitted diseases.Thus, it is an ambitious goal for the medical experts to seek an ideal blood substitutes.The ideal blood substitutes should replace both the volume expanding and oxygen carrier function of the homologous blood. The polymerized bovine hemoglobin solution was prepared using glutaraldehyde in the part 1 of this study. Hyperoxic solutions are the cheap and convenient oxygen carrier prepared by Chinese scientists. The effects of hyperoxia solutions in severe hemorrhagic shock were evaluated in the part 2 of the present study.Part 1 Study 1 OBJECTIVE Try to find out a suitable reaction conditions usingorthogonal design. METHODS Four factors including the mass of hemoglobin,the mass of glutaraldehyde,the reaction time and the pH of the Tris-HCl buffer solution were selected to be assigned in orthogonal design.Each factor was allocated 3 levels, the mass of hemoglobin: 10g,7.5g and 5g; the mass of glutaraldehyde: 6.25mg,12.5mg and 25mg;time: 20min,40min and 60min; pH of the Tris-HCl buffer solution: 7.2,7.6 and 8.0.Nine polymerizations were carried out by the orthogonal design of L9(34) table.The polymerized product was analysed by SDS-PAGE and the suitable reaction condition was screened.RESULTS Different polymerized products were acquired under different reaction conditions.The products were polymers composed of 16kD monomers and showed uncontinuous stick in SDS-PAGE. CONCLUSION The hemoglobin monomers can be effectively polymerized by glutaraldehyde. The suitable reaction condition was found by 7.5g hemoglobin, 12.5mg glutaraldehyde, 10mL Tris-HCl of pH 7.6 and 20min of polymerization using orthogonal design. Study 2OBJECTIVE The products acquired in study 1 were separated by gel filtration. Ultraviolet and visible spectroscopy and SDS-PAGE were applied to analyse the separated products. METHODS 7.5g hemoglobin and 12.5mg glutaraldehyde were polymerized 20min in 10mL Tris-HCl of pH 7.6.The polymerized products were separated by Sephadex G-100 gel filtration.The separated products were collected lmL/tube continuously.The products were analysed by spectroscopy and the absorbance of 220nm and 405nm were recorded.The products with similar absorbance were collected together and analysed by SDS-PAGE.RESULTS Obvious absorbed peak were appeared in both bovine hemoglobin monomers and polymerized hemoglobin solutions.But the absorbances in 220nm and 405nm of hemoglobin monomers were higher than polymerized hemoglobin solutions.The separated products with similar absorbance between 1-1.5 in 405nm were collected together.SDS-PAGE showed these products were mainly 64kD polymerizedproducts.CONCLUSION By gel filtration and spectroscopy, the64kD polymerized hemoglobin can be acquired.Part 2OBJECTIVE To investigate the effects of hyperoxic solutions on blood pressure and blood gas and survival rate during severe hemorrhagic shock in rabbits. METHODS Fourty-eight rabbits weighing (2.2-2.8 ) kg were randomly divided into two groups (n=24 each): hyperoxic solutions group(HO group) and control group.Each group was then randomly allocated to one of the four experimental groups: hydroxyethylstarch (HES, n= 6), polygenine (Pol, n = 6), gelofusine (Gel, n = 6) and Ringer's solution(RS,n = 6).Anesthesia was induced with 30mg mL-...
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