| To detect the guide line of supervise the acute rejection ,we made this kind of animal model. The success of this animal model determined the success of our experiment. In the process of making animal model we improved the the technique of donor cardiectomy , heparinization of recipient , preparative of celiac vessel , vascular anastomosis, myocardial perfusion and protection.,which made making heart transplantation animal model more convenient.Heart transplantation is a efficiency method in treating terminal heart diseases therapy in clinic. With the improvement of heart transplantation, the prevention and monitoring of acute rejection become the most difficult problem in medical area. In the past time ,many medical researchers studied the acute rejection just in immunology including antibody antigen cell factors and so on. But they did not do a deep research in the biology character about T cell. Our research detected the relation between the acute rejection and expression of CD28 molecular of T cell in heteroheart transplantation. After that we explored the the relation between the acute rejection and biological character of CD8+CD28+T. We attempted to exploit a new world about the research of the acute rejection.T cell is not a homogeneous group. Based on the expression CD4, CD8, we can divided T lymphocyte into CD4+T, CD8+T. CD4+T can be divided into Th and TDTH. CD8+T can be divided into Ts and CTL. Resting CTL presented in a preceding state. Recognized the antigen-MHC-I compound of the APC surface ,resting CTL was actived and became a effect cell against special antigen. CD28 molecule-a costimulated molecule was just expressed in surface of T cell.CD8+CD28+T was the main part of CTL.In part 3,our experiment detected the relation between the acute rejection and intracellular calcium, membrane potential and pH of the CD8+CD28+T.Our experiment detected the intracellular calcium membrane potential and pH of the CD8+CD28+T by fluorescence probe,which can combined with intracellular calcium hydrogen ion etc. The concentration of Fluo-3-AM fluorescence was positive correlation with the intracellular calcium. Through the concentration of BCECF-3-AM fluorescence we can know the concentration of hydrogen ion. DiBAC4 (3) was a sensitive fluorescence to the membrane potential.When more DiBAC4(3) entering the cell , the fluorescence became stronger,which shew the membrance depolarized stronger.Part 1 Animal Model of Heterotopic Heart Transplantation in ratsAbstract 1 Material and MethodMaterial 40 SPF grade male Wistar rats, 120 SPF grade male SD rats, weight 250-300g(offered by the experiment animal center of the first military medical university); operation appliance of microsurgery ( Shanghai medical appliance company); operation microscope(Shanghai medical optics appliance company);9/0 suture(Guangzhou medical appliance company)Operation Method:Anaesthesia by barbiturate(30mg/kg)1) Preparing of Recipient After anaesthesia the rat was on its back,fixed,epilation and sterilized. Through the abdominal incision(3-4cm) we shew the descending vena cava and abdominal aorta. The anastomosing position was choosed under the kidney artery. Recipient heparinized by 0.3mg heperine. Blocked the blood vessel ,we made a 2-3mm length incision on the blood vessel.we finished the preparing recipient.2) Donor Preparing Made the same abdominal incision , heparinized by 2mg heperine, droped the body temperature and then cut the two sides ribs and diaphragm, lifted the front chest wall, droped the heart temperature by ice water, cut the lung artery and arota and bundled the ascending and descending vena cava 1ung vein. Cut down the donor rat heart and perfused the donor heart by4CSt. Thomas saline water to make the heart muscle soft and pale.3 Heart Transplantation Anastomosed the aorta of the donor heart and the aorta of the recipient abdominal aorta and Anastomosed the pulmonary artery of the donor heart and the abdominal descending vena cava of the recipient. Expeled the gas of the blood vessel,loosened the desce... |
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