热缺血供肝耐受冷保存的时限研究

有研究表明，热缺血30min以内的供肝可考虑加以利用，本实验旨在探讨热缺血已达30min的供肝在UW液中冷保存的安全时限。     

供肝热缺血后对冷保存的耐受时限初步探讨

目的探讨供肝热缺血后耐受冷保存的安全时限。方法利用本组所建立的小型猪肝移植模型，设定供肝热缺血时间为20min，根据在UW液中的冷保存时间不同分为3组，分别冷保存12、16、20h，于肝移植术中及术后检测肝功能、肝脏病理、肝组织ATP含量、移植肝脏再灌注后微循环血流量及动物术后1周存活率。结果uw液冷保存12h组肝移植后小型猪1周内全部存活，而冷保存16、20h组存活率分别为20％、0％；随着冷保存时间的从12h延长到20h，ALT、AST逐渐上升，肝脏ATP含量、肝脏微循环血流量逐渐下降，形态学结果显示肝组织细胞变性、坏死及超微结构损害的程度逐渐加重。冷保存12h组与后两组上述指标存在显著性差异，生化及肝脏微循环指标的改变与病理结果及动物生存率相符合。结论在本实验条件下，热缺血时间为20min的供肝耐受冷保存的安全时限约为12h。     

自制CZ—1液对兔心肌组织低温延时保存的生化学指标观察

研制出我国自制的多种改良的ＵＷ液，是摆在移植界的一大课题。我院于１９９２年开始研制成长征－１号多器官保存液（简称ＣＺ－１液），同时对心肌组织０、６、１２、１８、２４小时冷冻保存后的生化学指标作了观察。结果表明：与ＵＷ液比较，ＣＺ－１液０、１２、１８、２４小时冷冻保存兔心肌组织ＡＴＰ含量无明显差别。而ＣＺ－１液冷冻保存心肌线粒体Ｃａ＾２＋超载明显缓于ＵＷ液。文章认为从生化指标变化来说，我们自的ＣＺ－     

热缺血和抗生素保存液对同种心瓣膜及主动脉活性的影响

研究不同热缺血时间对ＳＤ大鼠主动脉细胞活性的影响，以及应用改良抗生素液处理人尸主动脉及其瓣膜在４℃和－１９６℃下保存的细胞活性和组织结构的变化，结果显示，随着热缺血时间延长，尤其６ｈ以后，鼠主动脉细胞活性明显减低，经低浓度改良抗生素液处理，４℃保存６周和－１９６℃保存半年后，同种主动脉及瓣膜比对照组有更好的细胞活性和组织结构，更适合临床应用要求。     

不同冷保存时间的热缺血供肝在肝移植中的疗效

目的 探讨不同冷保存时间的热缺血供肝在肝移植中的疗效.方法 回顾性分析2006年1月至2007年12月中山大学附属第一医院收治的154例肝移植受者采用热缺血时间≤10 min的无心跳供者肝脏进行肝移植的疗效.根据冷保存时间将患者分为3组:<8 h为Ⅰ组,58例;8～12 h为Ⅱ组,62例;>12 h为Ⅲ组,34例.采用方差分析、t检验和X~2检验分析3组肝移植术后ALT峰值、并发症、移植肝存活和受者生存情况的差异.结果 3组受者术后均未发生原发性移植肝无功能.随访时间8～32个月,Ⅰ组受者的ALT峰值、感染发生率、胆道并发症发生率、移植肝存活率和生存率分别为(482±357)U/L、12%(7/58)、12%(7/58)、86%(50/58)和88%(51/58),Ⅲ组受者分别为(1274±608)U/L、29%(10/34)、26%(9/34)、68%(23/34)和71%(24/34),两组比较差异有统计学意义(t=5.X~2=4.28,6.77,4.51,4.28,P<0.05);而Ⅱ组受者仅ALT峰值达到(953±424)U/L,与Ⅰ组比较差异有统计学意义(t=4.76,P<0.05).结论 热缺血时间≤10 min的供肝能够耐受12 h的冷保存损伤,超过此时限,移植术后胆道并发症和感染的发生率显著升高,移植肝存活率和受者生存率显著降低.     

供肝热缺血后对冷保存的耐受时限初步探讨

目的探讨供肝热缺血后耐受冷保存的安全时限。方法利用本组所建立的小型猪肝移植模型,设定供肝热缺血时间为20min,根据在UW液中的冷保存时间不同分为3组,分别冷保存12、16、20h,于肝移植术中及术后检测肝功能、肝脏病理、肝组织ATP含量、移植肝脏再灌注后微循环血流量及动物术后1周存活率。结果UW液冷保存12h组肝移植后小型猪1周内全部存活,而冷保存16、20h组存活率分别为20%、0%;随着冷保存时间的从12h延长到20h,ALT、AST逐渐上升,肝脏ATP含量、肝脏微循环血流量逐渐下降,形态学结果显示肝组织细胞变性、坏死及超微结构损害的程度逐渐加重。冷保存12h组与后两组上述指标存在显著性差异,生化及肝脏微循环指标的改变与病理结果及动物生存率相符合。结论在本实验条件下,热缺血时间为20min的供肝耐受冷保存的安全时限约为12h。     

自制CMU-1液保存大鼠肝脏的实验研究

目的　探讨CMU 1液保存大鼠肝脏的效果。方法　根据灌注液和保存液的种类将Wistar大鼠分为两组 :UW组和CMU 1组 ,每组分 6h、12h、2 4h 3个保存时限 ,每亚组 6只大鼠。采用离体循环灌注模型 ,研究CMU 1保存液对保存肝脏能量代谢、生化功能、胆汁分泌及形态学方面的影响。结果　随着保存时间延长 ,肝组织TAN含量及AEC逐渐降低 ,CMU 1组较UW组下降略缓慢 ,保存 2 4h后高于UW组 (P <0 0 5 )。再灌注 12 0min后CMU 1组的肝脏分泌胆汁量较UW组多 (P <0 0 5 )。相同时限相比 ,灌出液中ALT、LDH值两组之间无显著差异 (P >0 0 5 )。肝脏组织学变化两组间无明显差异。保存 6h后 ,保存液pH值无明显变化 ;保存 12h后pH值下降 ,两组无明显差异 ;保存2 4h后 ,UW组pH值下降较CMU 1组明显。结论　CMU 1保存液保存大鼠肝脏效果与UW液相似 ,在改善保存肝脏能量代谢、预防细胞内酸中毒、胆汁分泌方面略优于UW液。     

热缺血后低温保存对供心的影响

目的　观察常温短暂热缺血后低温保存对供心的影响。方法　建立猪原位心脏移植模型 ,随机分为对照组 (C组 )和热缺血组 (H组 )。H组供心切取前常温缺血 5min。供心 4℃保存4h测定含水率、MDA、ATP含量 ,原位末端标记法检测心肌细胞凋亡 ;两组供心行原位心脏移植各8例 ,观察移植后血浆肌钙蛋白I(cTnI)水平、心排量 (CO )和再灌注 2h的心肌超微结构和凋亡。结果　与C组比较 ,H组供心低温保存后含水率、MDA高于C组 ,而ATP明显降低 ,差异均有显著性 (P <0 .0 5 ) ;C组cTnI漏出少 ,CO高于H组 (P <0 .0 5 ) ;H组心肌细胞结构改变明显 ,凋亡水平高于C组 (P <0 .0 1)。结论　热缺血使供心能量消耗 ,诱导再灌注后心肌细胞凋亡 ,一定程度上导致了供心移植后早期功能异常。     

对四种心肌保存液保存效果的评价

本文评价了4种心肌保存液的保存效果。结论是HTK液和UW液是良好的心肌保存液,尤其是HTK液保存效果优于UW液。Celsior液仅可用于心脏保存,对其它器官的保存效果还有待于进一步观察。Euro-Collins液中含有葡萄糖,使用复杂,现在几乎不再用于保存心肌。低温对器官保存十分重要。UW液在0～4℃条件下保存效果最好,HTK液在4℃和8℃时的保存效果无明显差别。     

前列腺素E1改善离体鼠心保存效果的研究

目的　探讨在冷缺血期供者心脏保存液中前列腺素 Ｅ１（ｐｒｏｓｔａｇｌａｎｄｉｎ Ｅ１）对改善鼠心的保存效果。　方法　将１４ 只大白鼠随机分为对照组和实验组，每组７ 只。对照组用 Ｓｔ Ｔｈｏｍ ａｓ停搏液停搏并保存大鼠心脏６ 小时；实验组在 Ｓｔ Ｔｈｏｍ ａｓ液中加入前列腺素 Ｅ１（５ μｇ／ Ｌ）停搏并冷保存６ 小时，利用离体鼠心非循环式 Ｌａｎｇｅｎｄｏｒｆｆ 灌流功能测定模型，测定左心室舒张期末压（ Ｌ Ｖ Ｅ Ｄ Ｐ）、左心室发展压（ Ｌ Ｖ Ｄ Ｐ）和左心室压力变化率（＋ ｄｐ／ｄｔ），并检测心肌细胞线粒体三磷酸腺苷（ Ａ Ｔ Ｐ）、腺嘌呤核苷酸总量（ Ｔ Ａ Ｎ）。　结果　实验组保存后的左心功能恢复明显优于对照组（ Ｐ＜ ０．０５），实验组保存后心肌细胞三磷酸腺苷和腺嘌呤核苷酸总量的浓度明显高于对照组（ Ｐ＜ ０．０５）。　结论　供者心脏停搏液和保存液中的前列腺素 Ｅ１ 能改善供心的保护效果。     

肺移植手术中供肺保存液的研究现状

肺移植手术是治疗终末期肺部疾病的惟一有效方法，但仍然有许多相关问题须待解决。除了供肺严重缺乏外，因缺血-再灌注损伤导致的移植肺功能异常是肺移植手术患者最常见的早期死亡原因之一。保存移植肺的最佳状态对减轻肺移植术后缺血器官功能障碍至关重要。因此，寻找一种高度可靠的肺保存液，对减轻移植肺的缺血-再灌注损伤、提高肺移植术后肺功能有着十分重要的意义。现就供肺保存液的种类、灌注方式、灌注条件及其改良措施的研究现状进行综述。     

Improved Preservation of Warm Ischemic Livers by Hypothermic Machine Perfusion with Supplemented University of Wisconsin Solution

Hypothermic machine perfusion (HMP) has the potential to improve recovery and preservation of Donation after Cardiac Death (DCD) livers, including uncontrolled DCD livers. However, current perfusion solutions lack the needed substrates to improve energy recovery and minimize hepatic injury, if warm ischemic time (WIT) is extended. This proof-of-concept study tested the hypothesis that the University of Wisconsin (UW) solution supplemented with anaplerotic substrates, calcium chloride, thromboxane A₂ inhibitor, and antioxidants could improve HMP preservation and minimize reperfusion injury of warm ischemic livers. Preflushed rat livers subjected to 60 min WIT were preserved for 5 h with standard UW or supplemented UW (SUW) solution. Post preservation hepatic functions and viability were assessed during isolated perfusion with Krebs–Henseleit solution. Livers preserved with SUW showed significantly (p <. 001) improved recovery of tissue ATP levels (μ mol/g liver), 2.06 ± 0.10 (mean ± SE), as compared to the UW group, 0.70 ± 0.10, and the level was 80% of that of fresh control livers (2.60 ± 0.13). At the end of 1 h of rewarming, lactate dehydrogenase (U/L) in the perfusate was significantly (p <. 05) lower in the SUW group (429 ± 58) as compared to ischemia–reperfusion (IR) (781 ± 12) and the UW group (1151 ± 83). Bile production (μ g/min/g liver) was significantly (p <. 05) higher in the SUW group (280 ± 13) as compared to the IR (224 ± 24) and the UW group (114 ± 14). The tissue edema formation assessed by tissue wet–dry ratio was significantly (p <. 05) higher in UW group. Histology showed well-preserved hepatic structure in the SUW group. In conclusion, this study suggests that HMP with SUW solution has the potential to restore and preserve livers with extended WIT.     

Three Preservation Solutions for Cold Storage of Heart Allografts: A Systematic Review and Meta‐Analysis

Organ preservation solution has been designed to attenuate the detrimental effects during the ischemic period. The aim of this study was to systematically evaluate the evidence comparing preservation solutions for heart preservation. Studies were searched in PubMed, Embase, the Cochrane Library, the Transplant Library, and the International Clinical Trials Registry Platform. The primary outcomes were patient survival and donor heart dysfunction. The secondary outcomes were in‐hospital mortality and enzyme gene expression. The University of Wisconsin solution (UW) was associated with a significantly improved survival at 30 days and 90 days (hazard ratio = 1.16, 95% confidence interval [CI] = 1.11–1.22, P < 0.00001; risk difference [RD] = 0.03, 95% CI = 0.01–0.05, P = 0.002), compared with Celsior. Hearts preserved with UW exhibited less ischemic necrosis than those preserved with Celsior (RD = ?0.07, 95% CI = ?0.08 to 0.05, P < 0.00001). UW was associated with better survival compared with histidine–tryptophan–ketoglutarate solution (HTK). There was no statistical difference in donor heart dysfunction and in‐hospital mortality outcomes when comparing HTK with Celsior solution. During static cold storage preservation, this study suggests that UW solution has better clinical outcomes for heart transplantation compared with the other two organ preservation solutions. Besides, the protective effect of Celsior solution is similar to HTK solution in donor heart preservation.     

Successful 24-h Preservation of Canine Small Bowel Using the Cavitary Two-layer (University of Wisconsin Solution/Perfluorochemical) Cold Storage Method

We previously developed the two-layer cold storage method (TLM), which allows sufficient oxygen delivery to the canine pancreas during preservation, and successfully achieved 96-h preservation. In this study, we applied a modified TLM (cavitary TLM) to small bowel preservation in a canine heterotopic transplant model. Using simple storage in University of Wisconsin solution (UWM, group 1, n = 12) or cavitary TLM (group 2, n = 8), 40 cm segments of the jejunum were preserved for 24 h at 4 degrees C. The nonpreservation group served as the control (group 3, n = 8). The grafts were implanted heterotopically as a Thiry-Vella loop. Eleven of 12 dogs in group 1 died within 3 days post-transplant as a result of graft intraluminal hemorrhage, while all dogs in groups 2 and 3 survived until day 7. Histological analyses showed almost normal structures of the graft mucosa in groups 2 and 3 at day 7. Results from maltose and acetaminophen absorption tests in group 2 were comparable to those in group 3. Only one survivor in group 1 showed distinct graft mucosal damage, confirmed by histological and functional analyses. In our transplant model, the canine small bowel was successfully preserved by cavitary TLM for at least 24 h, while this preservation time was beyond the limit with UWM.     

细胞内液型胶体液HBS对离体心脏的长期保存作用

探讨自制的含组氨酸、Ｂｉｍａｋａｌｉｍ的细胞内产体液对离体心脏的保存作用。方法：”健康性ＳＤ大鼠１６只，体重２４０－２６０ｇ。随机均分为两组，采用微量灌注技术，分别用ＨＢＳ和托马斯停搏液４℃保存离体鼠心２４小时，再灌注６０分钟进行比较研究。     

Limited Survival of Rat Small Bowel Transplants Preserved in University of Wisconsin Solution for 48 Hours

We have previously shown that rat small bowel may successfully be transplanted after preservation for 24 hours. In this study, syngeneic rat small bowel transplants were studied by light microscopy and transmission electron microscopy during and after preservation in University of Wisconsin (UW) solution for 48 hours. A total of 6 transplants were carried out using a previously described, standardized technique. In most cases, the bowel appeared histologically well preserved at the end of the 48 hr storage period (prior to implantation). Upon revascularization, however, reperfusion injury was dramatic, with loss of villi and crypts and inflammatory cells in all layers. The bowel was abnormal grossly as well as microscopically. This injury was irreversible with persistently abnormal histology for up to 1 week in all but 2 cases.

We conclude that UW solution alone may allow satisfactory preservation of intestinal grafts for 48 hours only in isolated cases, and is therefore not adequate for predictable, satisfactory 48 hr preservation. Attempts to prevent reperfusion injury with oxygen-free radical scavengers are in progress.     

24–Hour Preservation of Isolated Rat Hearts Perfused with Pyridoxalated Hemoglobin Polyoxyethylene Conjugate (PHP) Solution at Low Temperature

We examined the effects of long-term perfusion with pyridoxalated hemoglobin polyoxyethylene conjugate (PHP) solution on cardiac function of isolated rat hearts. Hearts were perfused with oxygenated Krebs-Henseleit (K-H) solution containing 3% PHP or hydroxyethyl starch (HES) at a constant pressure of 13 mm Hg for 24 h at 15 degrees C. After 24 h preservation, hearts were rewarmed with K-H solution. Heart rate (HR) in PHP-preserved hearts was almost the same as control and cardiac contractility was maintained at 70% of control, but coronary outflow decreased to about 50% of control. No edema developed in the PHP group. Addition of a Ca2+ antagonist, diltiazem, inhibited the elevation of the end-diastolic pressure significantly. In HES-preserved hearts, HR did not recover to control levels, but there was no significant difference in cardiac contractility between PHP- and HES-perfused hearts. The results demonstrate that isolated hearts can be preserved by perfusion with PHP solution under hypothermic conditions for 24 h and suggest that PHP solution would be useful for a perfusate of isolated organs and tissues for preservation.