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。     

Metabolism of prostaglandins in porcine liver transplantation with a graft harvested after 30- and 60-minute warm ischemia

The influence of warm ischemia on the metabolism of prostaglandins was investigated using a pig liver transplantation model employing the temporary portal arterialization technique. Eighteen pigs were divided into three groups according to warm ischemia time: 0 min (group I,n=6), 30 min (group II,n=6), and 60 min (group III,n=6). During portal arterialization, the hepatic venous prostaglandin E₂ (PGE₂) level in group III (3356.0±1011.8pg/ml) was significantly higher than that in group I (831.7±182.1pg/ml;P=0.0285). The hepatic venous PGE₂ levels were significantly higher than the arterial counterparts in all groups both at the beginning and during portal arterialization. At 60 min after portal revascularization, the arterial PGE₂ level in group III (886.7±268.0pg/ml) was significantly higher than that in groups I (99.0±18.6 pg/ml;P=0.0116) and II (204.2±65.4pg/ml;P=0.0282). Neither thromboxane B₂ (TXB₂) nor 6-keto PGF_1α showed any significant differences. In conclusion, the intra-operative changes of PGE₂ thus reflected the degree of warm ischemic damage, and PGE₂ could also be released from the graft. On the other hand, the increased levels of TXB₂ and 6-keto PGF_1α were throught to have an extrahepatic origin.     

Efficacy of prostaglandin I2 analog on liver grafts subjected to 30 minutes of warm ischemia

Due to the fact that no effective conditioning agent for hemodynamically unstable donors exists, the number of suitable donors is limited. The efficacy of OP-41483, a stable analog of prostaglandin I₂, as a conditioning agent of the liver was investigated in this study using pigs. OP-41483 was administered via the portal tributary for 10 min before 30 min of warm ischemia. Graft livers were procured after perfusion with OP-41483 in cold normal saline, the flushed with OP-41483 in Euro-Collins solution and placed in cold storage prior to orthotopic transplantation. OP-41483 was also administered intraportally for 120 min after reperfusion. Biochemical and histological studies, and survival rates were compared with a control group not given OP-41483 in an otherwise similar experimental protocol. The graft function recovered better in the OP group than in the control group, shown by the lactate values and lactate-to-pyruvate ratios. The marked congestion noted in the parenchyma of the control group livers was minimal in the OP group, verifying the microcirculatory effect of prostaglandin I₂ by its vasodilatory and antithrombotic actions. These findings suggest that OP-41483 has some protective effect as a conditioning agent in liver transplantation, with timing of administration being crucial.     

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.     

原位肝移植中移植肝早期功能的评价

目的通过测定原位肝移植中一些血清学指标来评价肝移植术后早期移植肝的功能状况。方法随机选取瑞金医院肝移植中心原位肝移植患者20例。分别在麻醉开始后（S）、无肝期开始即刻（A1）、无肝期结束前即刻（A2）、门静脉血流开放即刻（R0）、门静脉血流开放60min（R1）、门静脉血流开放3h（R3）、门静脉血流开放6h（R6）、门静脉血流开放12h（R12）、门静脉血流开放24h（R24）时间点抽取外周血，测定血清ALT、AST、LDH、GMP-140、vWF和HA。结果ALT、LDH在再灌注后即刻（R0）、AST在R6即达到峰值（P〈0．05），以后逐渐下降，但仍维持较高水平。LDH与ALT呈高度正相关（R=0．948，P〈0．001），LDH与AST呈中度正相关（R=0．646，P〈0．001）。GMP-140再灌注后开始升高，R3、R6、R12点达到峰值（P〈0．001），以后逐渐下降，但仍维持高水平。血管性血友病因子（von Willebrand factor，vWF）在再灌注后R6、R12、R24点高于正常。HA在再灌注后均高于正常。结论AST和ALT仍然是原位肝移植术后早期反映移植肝功能状况的可靠指标；LDH与ALT及AST存在良好相关性；GMP-140、vWF和HA的升高反映了肝窦内皮细胞（SEC）的损伤程度，但能否准确反映早期移植肝功能状况则有待进一步证实。     

Living related liver transplantation

Liver transplantation from a brain death donor has not yet been accepted in Japan. The only alternative method at present is transplantation from a living donor. After the first successful living related liver transplantation was performed by Strong in Brisbane, Australia, Japanese hepatic and transplant surgeons also began to perform such operations. As of February 1991, 16 living related liver transplantations had already been performed in Japan, mainly for children with biliary atresia. Five of these patients subsequently died, however, our patient has survived more than 1 year, and she is presently leading a normal school life. The most important issue regarding living related liver transplantation is to ensure the donor's safety. For this purpose, we conducted a preoperative banking of the donor's own blood and plasma. In addition, a selective vascular occlusion was carried out to reduce blood loss during the resection of the liver. Intraoperative color Doppler ultrasonography was introduced for evaluating the circulation of the graft. By using this modality, the following three points were able to be accurately estimated in order to obtain optimal graft perfusion: 1) The most suitable position for the graft to be fixed to the abdominal wall, 2) whether or not the abdominal wall could be closed and 3) the indication for a ligation of the collateral veins to form a porto-systemic shunt. Thanks to these procedures, living related liver transplantations have now become an acceptable transplant method, however, a transplantation from a cadaver that is brain dead but still has a beating heart is still absolutely necessary for adult recipients. Therefore, in the future, both methods should be performed.This report is the gist of a paper read at the 91st Annual Meeting of the Japanese Surgical Society, Kyoto, Japan, 1991     

冷缺血对大鼠小体积肝移植肝再生的影响

目的探讨冷缺血对大鼠小体积肝移植术后肝再生的影响.方法本组在2002年9月～2004年8月利用大鼠肝总量30%原位肝移植模型,实验分为肝总量70%肝切除组(C组)和冷缺血1、3、5 h组(E1、E2、E3组),观察1w生存率及肝重量/受体原肝重量比值(EGW/RLW),并检测术后1、2、3、7d肝细胞增殖活性及肝组织学变化.结果E1组1w生存率及EGW/RLW分别达100%和95%,均明显高于E2、E3组(P均＜0.05);E1、E 、E3组均于术后2d达到增殖高峰,其中E1组峰值显著高于E2、E3组(P＜0.001),且与C组峰值无差异(P＞0.05);组织学检查见E1组肝细胞核分裂明显活跃.结论冷缺血1 h的小体积供肝和70%肝切除后肝脏具有同样的增殖活性,仅增殖高峰稍晚;冷缺血超过3h严重影响小体积供肝的再生能力和受者的存活率.     

An evaluation of the tolerance of the autotransplanted canine lung against warm ischemia

Left lung autotransplantation was performed in mongrel dogs to examine the limitation of tolerance of the deflated lung against warm ischemia in a warm ischemic time (WIT) range of 60–360 minutes. The animals were divided into the 4 following groups according to the duration of WIT and the use of heparin: Group 1; WIT 60–120 min., heparin (−), Group 2; WIT 120–240 min., heparin (+), Group 3; WIT 240–360 min., heparin (+), and Group 4; WIT 240–360 min., heparin (−). All the Group 1 animals tolerated a right pulmonary artery ligation on the 6th postoperative day, but some of the Group 2 animals died immediately after the reimplantation due to pulmonary edema. All the Groups 3 and 4 animals died. The PaO₂ values during the right pulmonary artery occlusion, immediately after the reimplantation, correlated well with the survival of the animals. The maximum tolerable WIT of the deflated dog lung was considered to be 120 minutes.     

热缺血损伤对大鼠移植肝组织能量代谢及存活期的影响

目的：探讨不同热缺血时间下大鼠肝组织能量代谢变化规律,预测供肝耐受热缺血的安全时限。方法：实验动物按供肝热缺血时间分别为0、10、15、20、30、45和60min,随机分为7组。采用反相高效液相色谱法测定单纯热缺血后肝组织能量代谢指标并进行超微结构的观察。然后按各组条件分别作原位肝移植,观察移植后24、48h各组肝组织能量代谢指标的恢复性变化,并统计生存时间。结果：供肝经受热缺血损伤后,肝组织ATP含量和EC水平远逐渐下降,其中前30min下降比较急剧,以后趋向平缓。热缺血30min内肝组织ATP含量和EC水平在肝移植再复流24h后基本得到恢复,术后大鼠仍可以获长期存活。45min组,移植肝在48h后能量代谢的功能也基本恢复,虽不足以影响术后的1周存活率,但对大鼠肝移植术后的3个月存活率影响显著。60min组,肝脏能量代谢储备功能难以恢复,大鼠术后生存天数显著降低。结论：供肝组织三磷酸腺苷（ATP）含量和能荷（EC）水平以及其移植术后恢复的潜能是衡量供肝质量的重要标准。供肝热缺血损伤的时间与肝组织能量代谢功能的恢复及术后动物生存情况密切相关。     

先转流后游离肝脏的肝移植方法

目的 探讨先转流后游离肝脏的肝移植方法是否可行。方法 转流条件下的经典式肝移植23例（前期组）;进腹后先解剖肝十二指肠韧带、结扎切断胆总管及左右肝动脉、游离出门静脉进行转流,然后阻断肝上下腔静脉,游离肝周韧带及肝后下腔静脉,切肝、止血、植肝,共31例（后期组）。结果 转流时间:前期组（99.78±21.6）min,后期组（96.32±22.25）min;冷缺血时间:前期组（484.78±134.01）min,后期组（443.15±85.27）min;无肝期时间:前期组（100.00±21.19）min,后期组（102.00±24.31）min;术中失血量:前期组（5096.09±4243.59）ml,后期组（1726.23±1125.29）以P<0.01）;术中输血量:前期组（3676±2938.74）ml,后期组（1217.69±829.72）ml,两组相比 P<0.01。术后并发症:前期组（23例）26症例次,后期组（31例）19症例次。结论 先转流后游离肝脏的肝移植方法是一新的肝移植方法,有优越性。     

Intravital studies on beneficial effects of warm Ringer's lactate rinse in liver transplantation

This quantitative in vivo fluorescence microscopy study investigated the impact of warm versus cold Ringer's lactate (RL) graft rinse on various microvascular manifestations of ischemia-reperfusion injury after liver translantation in the rat. Syngeneic orthotopic liver transplantation, including arterial revascularization, was performed in male Lewis rats following 24 h of cold storage in University of Wisconsin (UW) solution. In one group (n=8) liver grafts were rinsed with 4°C (cold) RL, whereas in the other group (n=8) grafts were rinsed with 37°C (warm) RL immediately prior to revascularization. Hepatic microvascular perfusion, leukocyteendothelium interaction, and Kupffer cell activation were quantified 30–90 min after graft reperfusion by direct visualization with intravital fluorescence microscopy. Moreover, biliary excretory graft function was analyzed by determination of bile flow and bile salt excretion during the first 90 min after reperfusion. Compared to grafts rinsed with cold RL, acinar and sinusoidal perfusion were found to be significantly increased after rinsing the grafts with warm RL. The amount of nonperfused acini declined from 18.1%±4.0% to 7.4%±1.6% (P<0.05), and the total percentage of perfused sinusoids increased from 80.1±1.4 to 88.4±1.2 (P<0.001) after cold and warm rinse, respectively. After rinsing the graft with warm RL, WBC adherence in sinusoids and especially in postsinusoidal venules decreased significantly by 28% (P<0.001) and 33% (P<0.001), respectively. Kupffer cell activation was markedly reduced after rinsing with RL at 37°C, as indicated by a decelerated adherence of latex particles injected 80 min after reperfusion. Excretory graft function was dramatically increased following warm RL rinse during the 90-min observation period. Bile flow was enhanced from 1.04±0.5 to 3.9±0.8 ml/100 g liver per 90 min (P<0.01), with a parallel rise in bile salt excretion from 24.3±5.8 to 128.0±19.8 mmol/100 g liver per 90 min (P<0.05) when compared to cold RL. These data strongly suggest that rinsing liver grafts with warm RL prior to reperfusion represents a simple and inexpensive way to reduce the incidence of primary graft failure secondary to ischemia and reperfusion injury in liver transplantation.The results of this study were partly presented at the International Symposium on Transplantation of Abdominal Organs in Essen, Germany, in November 1992 and published in Transplantation Proceedings in 1993.     

Kupffer细胞在肝移植缺血再灌注损伤中的双重作用

Kupffer细胞足定居于肝内的巨细胞,在月十移植缺血再灌注损伤中发挥着重要的作用,门静脉恢复血流后刺激Kupffer细胞激活,释放活性氧族、多种炎性介质和细胞因子,对肝脏造成损伤.另一方面又可上调HO-1的表达,保护肝脏缺血再灌注损伤,因此,Kupffer细胞在肝移植缺血再灌注损伤中发挥着双重效应.     

Influence of cold ischemia time and graft transport distance on postoperative outcome in human liver transplantation

Purpose: The association between hepatic allograft cold ischemia time (CIT) and graft transport distance (GTD) in human liver transplantation was examined by investigating whether extended graft transportation prolongs the CIT and adversely affects graft survival. Methods: We retrospectively analyzed 186 consecutive orthotopic liver transplants (OLTs) done between May 1997 and July 1998. The number of miles from the donor hospital to the University of Pittsburgh Medical Center in a straight line was measured in each case, and defined as the GTD. The OLTs were divided into two groups according to whether the GTD was ≤200 miles or >200 miles. The latter group was then subdivided into groups of GTD 200–400 miles, GTD 400–600 miles, and GTD >600 miles. The CIT and graft outcome within 90 days after OLT were assessed. Results: Extended GTD prolonged the CIT (P < 0.001). The rate of hepatic allograft loss in the long GTD group was significantly higher than that in the short GTD group (P= 0.018). When the OLTs were subdivided according to GTD, the CIT increased and graft survival decreased as the GTD extended. Hepatic allograft transportation for a long distance prolonged the CIT and decreased the graft survival rate. Conclusion: Since prolonged CIT is a major risk factor, avoiding long-distance graft transportation is recommended when the donor risk factors are high. Received: September 6, 2001 / Accepted: March 5, 2002     

肝移植术中应用S-腺苷-L-蛋氨酸对供肝热缺血损伤的影响

目的探讨术中S-腺苷-L-蛋氨酸（SAMe）加入UW液和血浆冲洗液对热缺血损伤供肝及其恢复的影响。方法建立10min热缺血大鼠肝移植模型，分为A组：UW液灌注＋乳酸钠林格氏液冲洗、B组：UW液灌注＋血浆冲洗、C组：SAMe加入UW液灌注＋血浆冲洗和D组：UW液灌注＋SAMe加入血浆冲洗4组，观察肝组织组织病理学变化和电子显微镜下超微结构变化，并检测血清AST和透明质酸。结果C组和D组术后24h血清AST均低于B组（P〈0．05）。A组术后3h和24h血清HA高于B组（P〈0．05），B组复流后3h及24h血清HA均高于C组和D组（P〈0．05）。组织病理学表现B组复流后3h和24h肝细胞损伤和微循环紊乱较C组和D组明显；超微结构表现，A组复流后3h线粒体肿胀，肝窦内皮细胞肿胀，细胞核不规则，可见内皮细胞凋亡，大部分区域肝窦状隙明显狭窄，内皮层结构模糊，红细胞淤积，受压变形，白细胞附壁，可见内皮层完整性破坏；复流后24h，可见线粒体嵴断裂，核融解。B组内皮细胞损伤较A组轻，C组和D组超微结构表现微循环紊乱和肝细胞损伤表现较B组轻。结论供肝切取术中UW液中加入SAMe灌注保存，血浆冲洗液中加入SAMe可改善热缺血供肝微循环，减轻缺血再灌注损伤，并减轻肝细胞热缺血损伤，有利于10min热缺血供肝功能的恢复。     

乌司他丁减轻移植肾热缺血损伤的临床研究

目的　观察乌司他丁对移植肾热缺血损伤的保护作用。　方法　 86例肾移植患者根据供肾热缺血时间长短分为 2组 :A组 38例 ,热缺血时间 <5min ,B组 4 8例 ,热缺血时间 8～ 12min。其中A组 2 0例 ,B组 2 6例为乌司他丁治疗组 ,其余为对照组。观察术后第 1、3、7、10、14天血肌酐水平及肌酐恢复正常 ( <133μmol/L)的时间。 　结果　A组中治疗组术后第 1、3、7、10、14天血肌酐水平测定值分别为 :( 2 6 1.5± 14 3.8) ,( 137.7± 35 .3) ,( 131.3± 4 6 .2 ) ,( 12 6 .2± 32 .0 ) ,( 10 8.5± 2 8.6 )μmol/L ;对照组分别为 :( 32 0 .2± 15 6 .3) ,( 182 .2± 72 .9) ,( 139.8± 6 4 .1) ,( 133.1± 5 6 .4 ) ,( 116 .2±34.9) μmol/L ,2组比较差异无显著性意义 (P >0 .0 5 )。血肌酐恢复正常水平时间 :A组中治疗组为 2～ 12d ,平均 5 .8d ;对照组为 3～ 16d ,平均 7.2d ,2组比较差异无显著性意义 (P >0 .0 5 )。B组中治疗组术后第 1、3、7、10、14天血肌酐水平测定值分别为 :( 372 .6± 12 8.1) ,( 2 78.4± 38.9) ,( 14 5 .9±4 7.2 ) ,( 133.2± 39.8) ,( 12 8.0± 30 .6 ) μmol/L ;对照组分别为 :( 4 96 .3± 12 5 .6 ) ,( 36 4 .7±6 0 .2 ) ,( 196 .2± 36 .8) ,( 16 1.4± 4 1.5 ) ,( 14 9.8±     

Improving graft survival for patients undergoing liver transplantation

Dickson RC, Pungpapong S, Keaveny AP, Taner BC, Ghabril M, Aranda‐Michel J, Satyanarayana R, Bonatti H, Kramer DJ, Nguyen JH. Improving graft survival for patients undergoing liver transplantation.
Clin Transplant 2011: 25: E345–E355. © 2011 John Wiley & Sons A/S. Abstract: Liver transplant (LT) outcomes are reported to be improving in non‐HCV recipients but not for those infected with HCV. Our aim was to evaluate graft survival and predictors of outcome in HCV and non‐HCV patients before and after 2003. Patients with primary LT between February 1, 1998, and December 31, 2005, were included. Patients were divided into Era 1 (1998–2002) and Era 2 (2003–2005) with follow‐up through May 31, 2009. Graft survival was compared for HCV, non‐HCV, and all patients. There was significant improvement in graft survival in Era 2 for HCV patients. Graft survival in Era 2 of HCV patients was equivalent to non‐HCV patients. The most significant improvement between eras was in outcomes of grafts from donors ≥60 yr with three‐yr graft survival 58.6 (51.3–65.9) vs. 75.4 (68.9–81.9), p = 0.002. The use of donors ≥60 did not change between eras: 31% vs. 34%; however, utilization in HCV recipients decreased from 36% to 3% (p < 0.001). In conclusion, graft survival of HCV patients has improved significantly since 2003 and was comparable to non‐HCV patients up to three yr. The change in management of donor organs into HCV and non‐HCV patients likely contributed to this outcome.     

大鼠肝移植供肝热缺血时间与胆道损伤的关系

目的 探讨大鼠心源性死亡(DCD)供肝不同的热缺血时间与胆道缺血再灌注损伤的关系.方法 采用无肝素化的大鼠DCD供体肝移植模型,按供肝的热缺血时间分为0 min(WI0),10 min(WI10)、15 min(WI15)3组,每组36对大鼠.术后动态观察大鼠胆道病理改变及其并发症、肝功能指标,最后统计总体生存率.结果 供肝热缺血小于10 min时,术后胆道病理改变较轻且为可逆性改变,肝功能恢复较快;供肝热缺血时间为15 min时,术后胆道病理改变较重且为不可逆性改变,肝功能恢复延迟;3组胆道并发症的发生率差异有统计学意义(5.56%比8.33%比16.67%,P＜0.05).WI0组和WI10组的大鼠术后4周生存率差异无统计学意义(83.33%比77.78%,P＞0.05),而与WI15组比较4周生存率的差异有统计学意义(83.33%比58.33%,77.78%比58.33%,P＜0.05).结论 无肝素化的大鼠DCD供肝热缺血时间超过15 min时,移植术后胆道损伤明显,可导致不可逆改变.     

腹腔镜下肾部分切除术减少热缺血时间的研究进展

传统的腹腔镜肾部分切除术(LPN)为了减少术中出血、维持手术视野清晰是以夹闭肾蒂血管为标准的手术方式,由此导致的肾暂时缺血和再灌注损伤将不可避免的导致术后肾功能损害.随着外科技术的不断发展和各种新型止血材料的应用,腹腔镜下肾部分切除术正在不断的缩短热缺血时间,并向最终的目标“零缺血”迈进.本文就LPN在减少热缺血时间上应用的新技术予以综述.     

Evidence that both cyclosporin and azathioprine prevent warm ischemia reperfusion injury to the rat liver

The present work was undertaken to study whether the immunosuppressive agents cyclosporin (CyA) and azathioprine (AZA) ameliorate hepatic injury after warm ischemia. A temporary, normothermic liver ischemia was induced in female Sprague-Dawley rats. The rats were treated with CyA (10 mg/kg per day p.o.), AZA (8 mg/kg per day p.o.), or vehicles for 4 days before surgery. Seven-day survival rates after 60 min of ischemia improved significantly with CyA (76.2%, P<0.005) and AZA (78.6%, P<0.001) treatment, compared with 43.0% for the control group. The highest levels of serum aminotransferases in the treatment groups tended to be lower than those in the control group. The peak values for the percentage of liver necrosis, an indicator of the extent of hepatic necrosis, in the animals treated with CyA (26.1%±7.2%, mean±SEM) and AZA (32.1%±5.7%) were significantly lower than in the control group (47.4%±3.7%). Lipid peroxidative damage after reperfusion, assessed as the hepatic malondialdehyde (MDA) concentration, was significantly suppressed by pretreatment with CyA and AZA. Histological findings coincided with other parameters. This study demonstrates that both AZA and CyA have beneficial effects on normothermic liver ischemia in rats. It is suggested that the diminished lipid peroxidative damage with these agents might be one of the mechanisms responsible for this.