Fundamental Research Of Blood Transfusion Of Giant Panda

As one kind of rare animal that exclusively lives in China,Giant Panda was once on the edge of extinction,whose protection level has been regulated from endangered to easily endangered after years of protection.Medical treatment is a crucial part for the protection,and blood transfusion is an indispensable technique in the medical treatment.Most cases got cured by increasing hemoglobin content through blood transfusion,while one case of transfusion reaction and one case of death occurred(occurrence probability was about 5%),and 43% of 7 cases without timely blood transfusion died finally.Case above indicated that: firstly,transfusion of red blood cell(RBC)component is an effective way for the medical treatment;secondly,blood transfusion is a risky way due to lack of theoretical research.Systimetic rearch about the function and preservation and its immunohematological charactristcs of Giant Panda RBC would reduce the risk of adverse reactions of blood transfusion and be helpful for a more systimectic and scientific blood transfusion treatment which would be made a regular way to save more lives.RBC preservation is a prerequisite of blood transfusion.In this study,serious hemolysis developed within one week,if Giant Panda RBC samples were kept in solution for human RBC.The main reason for the phenomenon could be the difference of blood physiological characteristics between Giant Panda and human,so pH value,plasma osmotic pressure,erythrocyte fragility and blood gas analysis were tested and preserving solution(4?)for Giant Panda were formulated.It turned out that red blood cell of giant panda could be preserved effectively and successfully for about 30 days.Immunological characteristics of Giant Panda RBC is the crucial factor for its transfusion,and there was no systematic rearch about the immunological characteristics of Giant Panda and Asiatic Black Bear RBC.So,the compatibility among different Giant Panda individuals and the compatibility between Giant Panda and Asiatic Black Bear were explored and immunohematological study of Giant Panda and Asiatic Black Bear RBC were conducted in which human blood typing reagents,which were corresponding to the conservative sequence of the mammal RBC membrane protein,were used.In Giant Panda blood compatibility tests(blood cross matching tests,BCMT),weak agglutination phenomena among different blood samples(n=17)were observed and the agglutination enhanced at 4?,and the compatible rate among different individuals was 71.11% at the temperature.In Asiatic Black Bear blood compatibility tests(n=40),agglutination phenomena occurred between 5 samples and the others in three serological patterns.Xenogeneic agglutination phenomena between Giant Panda(n=17)and Asiatic Black Bear(n=10)blood samples were also observed.Results above all indicated that Giant Panda have suspected blood group antigen(s)and there were at least three kinds of suspected blood group antigens in Asiatic Black Bear blood.Human serological typing reagents were used in further study about the suspected blood group antigen(s)of Giand Panda and Asiatic Black Bear RBC,and natural serum of human and mice were used as control.It came to the conclusions: first,in the study we found that the interference of interspecific antibodies could not be avoided in BCMT between human originated anti-serum and Giant Panda RBC,so the agglutination between human originated anti-M serum and Giant Panda RBC,which indicated that Giant Panda had M antigen(Diao Yuying,1989),might be caused by interspecific antibodies rather than the reaction between blood group antigen and antibody;Second,ABO blood group antigens were not found in Giant Panda by using human ABO blood reagents,which was different from former reports that the Giant Panda RBC was similar to human O-type RBC(Wang Dechun,1999);Third,human anti-B antibody could be used for the blood screening of Asian Black Bear.The erythrocyte membrane protein which determined the characteristics of the functional and immunogenic characteristics of RBC,so molecular biological analysis of the Giant Panda erythrocyte membrane protein was done in this study.In the former study,it was found that compared with Asiatic Black Bear,Giant Panda has less hematocrit and fewer RBC counting.The Giant Panda and Asiatic Black Bear are Ursidae animals and have similar size,how does Giant Panda use less amount of RBC to maintain body metabolism? In further study,liquid chromatography with tandem mass spectrometry(LC-MS/MS)analysis was done after the purification of Giant Panda RBC and 388 kinds of membrane proteins were found in Giant Panda and 315 in Asiatic Black Bear.Finally,in the comparison of LC-MS/MS results between Giant Panda and Asiatic Black Bear,it was found that the Giant Panda exerted more special enzymes and other proteins in glycolysis than Asiatic Black Bear,which were Hexokinase X2,Glyceraldehyde 3-phosphate Dehydrogenase,Acetyl coenzyme A synthase 2,and ATP-binding cassette protein(ABC transporter),including ABCB6,ABCB8 and ABCC1,which meant higher efficiency level in RBC metabolism.It was also found that Giant Panda exclusive membrane protein took part in O2/CO2 exchange as well as metabolism,which made it possible that Giant Panda could exchange O2/CO2 with less RBC and also revealed the unique function of Giant Panda RBC and its blood physiological characteristics of less hematocrit and fewer RBC counting caused by the function.To validate the mass spectrum results,gene sequence(SLC4A1)of Giant Panda band3 protein was amplified and cloned.Exsistence of Giant Panda band3 protein was proved by analyzing its sequence and functional domain,which verified the accuracy of LC-MS/MS.Function of Giant Panda band3 protein was analyzed by its functional domain,and the evolution of Giant Panda SLC4A1 gene was analyzed.In conclusion,blood transfusion is an important way for saving Giant Panda,but the research on Giant Panda blood transfusion system was deficient.In this study,experiments were designed according to the key point of Giant Panda blood transfusion and situations of blood sample insufficiency,and blood preserving was set as the threshold and foundation for later study.Immunohematological characteristics of Giand Panda and Asiatic Black Bear RBC were analyzed and RBC physiological characteristics and metabolisim were compared with LC-MS/MS,which laid the foundation for further rearch of function and membrane protein of Giant Panda RBC and also provided theoretical basis for blood transfusion.