Organ Transplantation from non-heart beating

1. 根据我们之前的犬类实验，胰腺移植物经过90 min的热缺血后，未经处理不能存活，但两层法保存5小时后可以成功移植。假设两层保存期间的蛋白质合成与移植物复苏有关，进行了以下实验。采用双层法（全氟化学/UW溶液）在20℃下保存90 min热缺血损伤的犬胰腺5小时。通过测定在保存液中分别加入的氚化胸腺嘧啶、氚化尿嘧啶和氚化亮氨酸的掺入量，定量保存过程中菌落内DNA、RNA和蛋白质的合成。通过自体颈椎皮下腔移植后的移植物存活情况来判断移植物的生存能力。7个移植物在热缺血后立即移植均无成活（0%），而在20℃下两层保存5小时后移植的7个移植物全部成活（100%）。在双层法保存期间，观察到RNA和随后的蛋白质合成显著增加，而不是通过UW简单冷藏。相比之下，DNA的合成没有跟随RNA和蛋白质的合成。我们认为，在两层法的低温保存下，蛋白质的合成可能与胰腺移植术后细胞的恢复过程有关。一些报道表明，热休克蛋白（HSPs）在细胞恢复中具有重要功能。采用双层保存法研究热休克蛋白在减少缺血再灌注损伤中的作用。免疫组化检查和热休克蛋白的western-blotting结果显示，热休克蛋白60可能与热缺血损伤胰腺的功能恢复有关。

1. Based on our previous canine experiments, pancreas grafts subjected to 90 min warm ischemia, which cannot survive after implantation with no treatment, could be successfully transplanted after 5-hour preservation by the two-layer method. On the hypothesis that protein synthesis during the two-layer preservation would be associated with this graft resuscitation, the following experiments were undertaken. We preserved canine pancreases damaged by 90-min warm ischemia using the two-layer method (perfluorochemical/UW solution) for 5 hrs at 20℃. Intragraft DNA, RNA and protein synthesis were quantitated durins preservation by determining incorporation of tritiated thymidine, tritiated uridine and tritiated leucine, respctively, added in this preservation solution. Graft viability was judged from graft survival after autotransplantation in the cervical subcutaneous cavity. None of 7 grafts (0%) survived when they were immediately transplanted after warm ischemia, while all of 7 grafts (100%) transplanted after the two-layer preservation for 5 hrs at 20℃ survived. Significant increases in RNA and subsequent protein syntheses were observed during preservation by the two-layer method, not by UW simple cold storage. In contrast, DNA synthesis did not follow RNA and protein synthesis. We concluded that protein synthesis might be associated with the process of postischemic cellular recovery of the pancreas graft during mild hypothermic preservation by the two-layer method.2. Some reports have indicated that heat shock proteins (HSPs) have important functions in cellular recovery. We studied the role of HSPs in reducing ischemia-reperfusion injury during preservation by the two-layer method. From the results of immunohistochemical examinations and western-blotting of HSPs, HSP-60 was likely associated with functional recovery of warm ischemically damaged pancreas by the two-layer method.