Experimental study of the compound tissue preservation

(1)采用新鲜血液体外循环系统对大鼠后肢新鲜断肢和后肢在4或1 C条件下保存12、24、48和72 h的血管顺应性和血管平滑肌收缩性进行评价。与新鲜断肢相比，-1 C贮藏12 h以上血管顺应性明显降低（p<0.01）。血管平滑肌在-1 C下保存24 h后，其收缩力与新鲜断肢无明显差异。然而，在48小时或更长时间后，它明显下降。将这些结果与4 C保存的四肢进行比较，结果表明-1 C的血管顺应性不如4 C，但在平滑肌收缩性方面优于4 C。(2)观察低温在义肢移植中的应用。平均动脉血流量显著降低44%。耗氧量显著下降了常温值的28%。(3)采用northern blot法定量检测大鼠胫骨前肌组织中甘油醛3-磷酸脱氢酶（GAPDH）的mrna表达。将切除后肢分为常温（22ºC）保存0、1、3、6、9 h和常温（4ºC）保存6、9 h 7组。在预先确定的缺血时间后，从后肢切除每块肌肉并分析mRNA。在对照组中，GAPDH条带明显检测到。然而，在常温保存组中涂抹条带。室温保存3小时或更长时间后，mRNA含量显著低于新鲜截肢肌肉。在4ºC环境下保存9 h后，mRNA没有明显减少。这些结果表明GAPDH的mRNA降解与肌肉损伤相关，因为肌肉损伤随着保存时间的推移而增加。

(1)The vascular compliance and the contractility of vascular smooth muscle of rat freshly amputated hindlimbs and hindlimbs preserved at 4ﾟC or-1ﾟC for 12,24,48, and 72 hours were evaluated using a fresh blood extracorporeal circulatory system. Vascular compliance after storage at -1ﾟC for 12 hours or longer decreased significantly from that of freshly amputated limbs (p<0.01). The contractility of vascular smooth muscle after storage at -1ﾟC for up to 24 hours were not different from that of freshly amputated limbs. However, it decreased significantly after 48 hours or longer. A comparison of these results to limbs Preserved at 4ﾟC suggested that the storage at -1ﾟC was inferior to that at 4ﾟC with respect to the vascular compliance, but was superior to the storage at 4ﾟC with respect to smooth muscle contractility. (2)The application of hypothermia to the replanted limb was observed. The mean arterial blood flow decreased significantly by 44% with cooling at 25ﾟC.The oxygen consumption decreased significantly by 28% of normothermia values. (3)Messenger RNA for GAPDH (glyceraldehyde 3-phosphate dehydrogenase) on the preserved anterior tibial muscle of rats were evaluated quantitatively using northen blot analyzes. The amputated hindlimbs were divided into 7 groups : 0,1,3,6,9 hours storage at room temperature (22ﾟC) and 6,9 hours storage at 4ﾟC.After a predetermined period of ischemia, each muscle was resected from the hindlimb and mRNA was analyzed. In the control group, the band for GAPDH was clearly detected. However, the bands were smeared in normothermic preserved groups. After 3 or more hours of storage at room temperature, mRNA content was significantly less than freshly amputated muscle. After up to 9 hours storage at 4ﾟC,mRNA did not decrease markedly. These results indicated that mRNA degradation of GAPDH correlate with muscle damage, since muscle damage was increased as preserved time passed.