异种肝细胞移植防治大鼠急性肝功能衰竭

目的: 探讨异种肝细胞移植对大鼠急性肝功能衰竭的治疗效果. 方法: 切除大鼠90%肝脏,建立急性肝功能衰竭模型.分离异种豚鼠和同种异体Sprague-Dawley鼠肝细胞,切肝术前1天植入受体脾脏内.观察受试大鼠存活时间、切肝术后24小时血生化改变和脾脏切片表现. 结果: ①中位存活时间,对照组为21小时,同种组为56小时,异种组为40小时.同种组存活时间较对照组长(P<0.01),异种组亦延长(P<0.05).②移植组部分血生化指标有所改善.异种组中仅血糖和凝血酶原时间改善较明显(P<0.05),同种组中谷丙转氨酶、总胆红素、血糖和凝血酶原时间都有明显改善(P<0.05或P<0.01).③肝细胞植入后48小时,异种组脾脏内仅存少量萎缩的肝细胞,同种组脾脏内仍散在健存肝细胞. 结论: 异种肝细胞移植对大鼠急性肝功能衰竭有防治作用;要提高移植效果,首先需克服早期的免疫排斥.     

微囊化罗非鱼肝细胞移植治疗大鼠急性肝功能衰竭

目的 观察微囊化罗非鱼肝细胞移植对急性肝衰大鼠的治疗作用.方法 D-氨基半乳糖制备的肝衰大鼠随机分为4组:微囊化组,裸肝细胞组、空微囊组及生理盐水组,腹腔内分别植入微囊化罗非鱼肝细胞、裸肝细胞、空微囊及生理盐水.比较各组间1周死亡率.移植后动态检测各组大鼠总胆红素,比较其差异,动态观察移植物的病理变化.结果 移植后1周内微囊化组的存活率较高(57.9%),与其他各组比较差异有统计学意义(P＜0.05).移植后48 h,微囊化组总胆红素(6.21±0.86)μmol/L明显较低,与其他各组差异有统计学意义(P＜0.01).移植后1周,微囊化组可以收集到形态完整,没有粘连的微囊,其内的肝细胞尚部分保持存活.结论 微囊化罗非鱼肝细胞移植能改善肝衰大鼠的肝功能、提高存活率.     

肝细胞移植治疗急性肝功能衰竭的研究进展(文献综述)

明确“肝细胞移植治疗急性肝功能衰竭”领域中的概念与治疗方法具有重要意义 ,本文就肝细胞的紧急辅助移植治疗肝功能衰竭的原理、移植物的制备、病例选择以及手术后监测等问题给予综述     

肝细胞移植治疗急性肝功能衰竭的研究进展（文献综述）

明确“肝细胞移植治疗急性肝功能衰竭”领域中的概念与治疗方法具有重要意义，本文就肝细胞的紧急辅助移植治疗肝功能衰竭的原理、移植物的制备、病例选择以及手术后监测等问题给予综述。     

急性肝功能衰竭肝细胞移植后的肝再生研究进展

急性肝功能衰竭（acute liver failure，ALF）的治疗十分棘手。自1985年开始采用急诊行原位肝移植治疗以来，术后一年生存率已提高至83％。但是急诊行原位肝移植存在供体短缺等缺点，其应用受到一定限制。近年肝细胞移植的研究取得很大进展。有望成为治疗ALF的另一种有效手段。由于肝脏的再生潜力很大．移植肝细胞所提供的暂时、有效的肝功能支持．为受体肝脏的再生恢复赢得了时间，从而可使ALF病人有时间等待肝移植，甚至有可能避免肝移植。研究ALF肝细胞移植后的肝再生及其机制．探讨其影响因素．对肝细胞移植的临床应用具有重要的意义。本文就有关ALF肝细胞移植后肝再生的研究进展综述如下。     

异种肝细胞移植缓解大鼠急性肝功能衰竭及其排斥反应观察

目的探讨异种肝细胞移植对大鼠急性肝功能衰竭防治效果及其免疫排斥反应。方法将异种豚鼠肝细胞,90%肝除术前1d植入大鼠脾脏内。观察受试大鼠存活时间,及切肝术后24h血生化改变。另外观察植入肝细胞被排斥情况及受体CH50、异种抗体IgG、IgM水平变化。结果(1)中位存活时间,对照组为21h,同种组为56h,异种组为40h。同种组存活时间较对照组延长(P<0·01),异种组亦延长(P<0·05)。(2)异种组血糖和凝血酶原时间改善较明显(P<0·05),同种组谷丙转氨酶、总胆红素、血糖、凝血酶原时间都有明显改善(P<0·05或P<0·01)。(3)受体CH50和异种抗体IgM水平下降与植入异种肝细胞排斥过程同步。结论异种肝细胞移植对大鼠急性肝功能衰竭有防治作用,补体和异种抗体IgM与排斥反应关系密切。     

肝细胞移植治疗大鼠急性药物性肝功能衰竭的疗效观察

肝细胞移植治疗急性肝脏功能衰竭、肝脏先天性代谢性等疾病正成为器官移植学的热点。本实验目的是探索机械法分离肝细胞并对新鲜分离肝细胞移植治疗急性肝脏功能衰竭模型的效果进行评价。实验材料与方法1.实验动物 :供体为纯系雄性SD大鼠 ,鼠龄为4～ 5个月 ,体重为 2 5 0～ 30 0g。受体为SD雄性大鼠 ,鼠龄为 5～ 6个月 ,体重为 30 0～ 35 0 g。2 .试剂和仪器 :D 氨基半乳糖、TypanBlue(苔芬兰 )试剂、OLYMPUS倒置AH 2型显微镜、BECKMANSYNCHRONCX4CE型全自动生化分析仪。3.建立急性药物性肝功能衰竭模型 :大鼠腹腔内注射 10 %…     

大鼠肝细胞肾脏内移植后组织形态与肝功能衰竭治疗作用研究

本实验通过同种异体大鼠肝细胞肾脏内移植，尝试在受体体内建立第二肝脏。对移植肝细胞进行长期组织形态与功能观察，并利用该方法进行急性肝功能衰竭肝功能辅助支持治疗研究。结果表明肝细胞肾脏内移植后可长期存活、增殖而且具有肝组织结构重建及保持肝脏功能的能力。４×１０７个肝细胞肾脏内移植后可显著提高受体大鼠对肝功能衰竭的承受力，移植组术后３天行９０％肝切除诱发急性外科型肝功能衰竭后其存活率显著高于对照组。以上结果显示肾脏可以成为肝细胞移植又一适宜部位，肝细胞肾脏内移植有可能成为临床上肝脏代谢及功能障碍疾病的一种治疗手段。     

异种肝细胞移植排斥机理的探讨

目的：探讨异种肝细胞排斥反应的机理,为治疗异种肝细胞移植排斥反应提供理论依据。方法,用D－氨基半乳糖腹腔内注射制成肝功能衰竭大鼠模型,并在其脾内植入豚鼠肝细胞,通过免疫组化方法,用抗大鼠IgM抗体和抗大鼠IgG抗体,抗大鼠CD4和抗大鼠CD8抗体与因排斥反应在大鼠脾 内可能产生的IgM,IgG抗体和CD4,CD8淋巴细胞结合,在移植后不同时间取受鼠脾脏标本,检测其内是否有IgM,IgG抗体和CD4,CD8淋巴细胞存在。结果：移植后12hIgM抗体存在,移植后24h大鼠脾内出现IgG抗体,同时有CD4^=和CD8^＋淋巴细胞,且在移植后1周大鼠脾内均可见上述4种物质。结论：体液和细胞免疫均参与异种肝细胞移植的排斥反应。     

应用环孢素A治疗异种肝细胞移植的排斥反应

目的　探讨应用环孢素A(CsA)治疗异种肝细胞移植排斥反应的疗效及其排斥机制。方法　将豚鼠肝细胞植入D-氨基半乳糖诱导的肝衰大鼠脾内,分CsA治疗组和单纯移植组。观察2周生存率及脾病理组织学检查;酶联免疫双抗体夹心法(Sandwich-ELISA)测定大鼠血清白细胞介素(IL)-2的浓度。结果　2周生存率比较CsA治疗组（78.6%）与单纯移植组（80.0%）相比差异无显著性(P＞0.05)。应用CsA可减缓炎症细胞浸润。血清IL-2浓度检测正常大鼠为(28.7±7.0)ng/L,移植后各组血清IL-2浓度变化不大,直至移植后第7天,CsA治疗组为(28.5±6.0)ng/L、单纯移植组为(31.5±8.0)ng/L。3组间差异均无显著性(P＞0.05)。结论　在异种肝细胞脾内移植中,细胞免疫发生的较迟和/或较弱。单独应用CsA治疗异种肝细胞脾内移植引发的排斥反应其疗效不佳。     

肝移植术后一过性肝内胆汁淤积症与急性排斥反应的鉴别

目的探讨肝移植术后一过性肝内胆汁淤积症（transient intrahepatic cholestasis，TIHC）与急性排斥反应（acute rejection，AR）的鉴别方法。方法按诊断标准将30例肝移植患者分为TIHC组与AR组，观察比较2组的谷丙转氨酶（ALT）及直接胆红素（DBIL）在术前1d，术后早期（术后3d、7d及21d）的变化特点。结果术后第7d对比术后第3d，TIHC组DBIL明显升高而ALT无明显变化；AR组DBIL明显升高同时ALT呈升高趋势。经过针对性治疗，至术后第21d，2组ALT及DBIL均明显下降。结论肝移植术后DBIL及ALT的变化特点对于TIHC与AR的鉴别有一定意义。     

Isolated Donor Specific Alloantibody-Mediated Rejection after ABO Compatible Liver Transplantation

Antibody-mediated rejection (AMR) after liver transplantation is recognized in ABO incompatible and xeno-transplantation, but its role after ABO compatible liver transplantation is controversial. We report a case of ABO compatible liver transplantation that demonstrated clinical, serological and histological signs of AMR without evidence of concurrent acute cellular rejection. AMR with persistently high titers of circulating donor specific antibodies resulted in graft injury with initial centrilobular hepatocyte necrosis, fibroedematous portal expansion mimicking biliary tract outflow obstruction, ultimately resulting in extensive bridging fibrosis. Immunofluorescence microscopy demonstrated persistent, diffuse linear C4d deposits along sinusoids and central veins. Despite intense therapeutic intervention including plasmapheresis, IVIG and rituximab, AMR led to graft failure. We present evidence that an antibody-mediated alloresponse to an ABO compatible liver graft can cause significant graft injury independent of acute cellular rejection. AMR shows distinct histologic changes including a characteristic staining profile for C4d.     

肝移植术后急性肺损伤的观察与分析

目的 分析肝脏移植患者术后并发急性肺损伤(AU)的危险因素，并探讨其防治措施。方法 采用回顾性调查的方法，对4例肝脏移植患者的相关资料进行分析。结果 门肺高压症、呼吸机长期使用、严重感染、全身炎性反应综合征、高凝状态、液体超载、肾功能障碍等．是肝移植术后并发急性肺损伤及急性呼吸窘迫综合征(ARDS)的危险因素。结论 肝移植术后易发生ALI，预防并减少相关危险因素的影响．对减少术后ALI及ARDS有重要意义。     

Bortezomib for Acute Antibody‐Mediated Rejection in Liver Transplantation

Antibody‐mediated rejection (AMR) is an uncommon, but challenging type of rejection after solid organ transplantation. We review three cases of AMR in ABO‐compatible liver transplant recipients. These cases were characterized by severe acute rejection resistant to steroids and antithymocyte globulin, histologic evidence of plasma cell infiltrates, C4d positivity and high serum anti‐HLA donor‐specific antibodies. All three patients were treated with bortezomib, a proteasome inhibitor effective in depleting plasma cells. After treatment, all patients had improved or normal liver function tests, resolution of C4d deposition and significant decline in their HLA donor‐specific antibodies.     

Risk Factors for Development of Acute Renal Failure After Liver Transplantation

Background.?Acute renal failure (ARF) is a common complication after liver transplantation (LTx). Identification of risk factors may prevent the development and attenuate the impact of ARF on patients outcome after LTX. Methods.?Retrospective analysis of variables in the pre, intra, and postoperative periods of 92 patients submitted to LTx was performed in order to identify risk factors for development of ARF after LTx. ARF was defined as serum creatinine ≥2.0 mg/dL in the first 30 days after LTx. Univariate and multivariate analysis by logistic regression were performed. Results.?ARF group comprised 56 patients (61%). Preoperative serum creatinine was higher in ARF group. During the intraoperative period, ARF group required more blood transfusions, developed more episodes of hypotension and presented longer anesthesia time. In the postoperative period, ARF group presented higher serum bilirubin and more episodes of hypotension. Dialysis was required in 10 patients (11%). The identified risk factors for development of ARF were: preoperative serum creatinine >1.0 mg/dL, more than five blood transfusions in the intraoperative period, hypotension during intra and postoperative periods. The identified mortality risk factors were hypotension in the postoperative period and no recovery of renal function after 30 days. Conclusions.?Several factors are involved in the pathogenesis of ARF after LTx and may influence patients outcome and mortality. Pretransplant renal function and hemodynamic conditions in the operative and postoperative periods were identified as risk factors for development of ARF after LTx. Nonrenal function recovery and postoperative hypotension were identified as mortality risk factors after LTx.     

Risk Factors for Rejection and Infection in Pediatric Liver Transplantation

Rejection and infection are important adverse events after pediatric liver transplantation, not previously subject to concurrent risk analysis. Of 2291 children (<18 years), rejection occurred at least once in 46%, serious bacterial/fungal or viral infections in 52%. Infection caused more deaths than rejection (5.5% vs. 0.6% of patients, p < 0.001). Early rejection (<6 month) did not contribute to mortality or graft failure. Recurrent/chronic rejection was a risk in graft failure, but led to retransplant in only 1.6% of first grafts. Multivariate predictors of bacterial/fungal infection included recipient age (highest in infants), race, donor organ variants, bilirubin, anhepatic time, cyclosporin (vs. tacrolimus) and era of transplant (before 2002 vs. after 2002); serious viral infection predictors included donor organ variants, rejection, Epstein-Barr Virus (EBV) naivety and era; for rejection, predictors included age (lowest in infants), primary diagnosis, donor-recipient blood type mismatch, the use of cyclosporin (vs. tacrolimus), no induction and era. In pediatric liver transplantation, infection risk far exceeds that of rejection, which causes limited harm to the patient or graft, particularly in infants. Aggressive infection control, attention to modifiable factors such as pretransplant nutrition and donor organ options and rigorous age-specific review of the risk/benefit of choice and intensity of immunosuppressive regimes is warranted.     

A Peripheral Blood Diagnostic Test for Acute Rejection in Renal Transplantation

Monitoring of renal graft status through peripheral blood (PB) rather than invasive biopsy is important as it will lessen the risk of infection and other stresses, while reducing the costs of rejection diagnosis. Blood gene biomarker panels were discovered by microarrays at a single center and subsequently validated and cross‐validated by QPCR in the NIH SNSO1 randomized study from 12 US pediatric transplant programs. A total of 367 unique human PB samples, each paired with a graft biopsy for centralized, blinded phenotype classification, were analyzed (115 acute rejection (AR), 180 stable and 72 other causes of graft injury). Of the differentially expressed genes by microarray, Q‐PCR analysis of a five gene‐set (DUSP1, PBEF1, PSEN1, MAPK9 and NKTR) classified AR with high accuracy. A logistic regression model was built on independent training‐set (n = 47) and validated on independent test‐set (n = 198)samples, discriminating AR from STA with 91% sensitivity and 94% specificity and AR from all other non‐AR phenotypes with 91% sensitivity and 90% specificity. The 5‐gene set can diagnose AR potentially avoiding the need for invasive renal biopsy. These data support the conduct of a prospective study to validate the clinical predictive utility of this diagnostic tool.     

Induction of Rejection After Small-for-Size Liver Transplantation: Size Matters

Background: Reduced-size liver transplantation is associated with liver regeneration. This study was designed to analyze the influence of graft size on liver rejection and liver regeneration. Methods: Reduced-size liver transplantations were performed in the rejecting ACI to Lewis and the graft acceptance BN to Lewis strain combination. The BN to Lewis control group was treated with the immunosuppressive drug FK506. Results: An accelerated liver rejection in the ACI to Lewis strain combination was found in small-for-size partial liver grafts. Graft weight to recipient liver weight ratio (GW/RLW) showed a positive correlation with survival time. In the BN to Lewis strain combination, lethal rejection was seen in small-for-size partial liver grafts. A critical immunologic GW/RLW of 33% was calculated. In rats dying from lethal rejection, GW/RLW and survival time showed a positive correlation. However, GW/RLW showed a negative correlation with hepatocellular proliferation. In regenerating livers, MHC II upregulation was also observed in the control group. All control animals survived small-for-size liver transplantation. Conclusions: The relative graft size seems to be a decisive factor influencing the kinetic of liver rejection and the induction of liver rejection. Relative critical immunologic liver mass was determined to be 33%.     

Over-Expression of AIF-1 in Liver Allografts and Peripheral Blood Correlates with Acute Rejection after Transplantation in Rats

Early and accurate detection of acute cellular rejection (ACR) is important in the management of liver allograft recipients. We hypothesized that expression of allograft inflammatory factor (AIF)-1 would be associated with liver allograft rejection as previous studies have shown that a relationship exists between kidney and heart transplantation. Indeed using rat orthotopic transplant models we found that the expression of allograft inflammatory factor-1 (AIF-1) can be detected in both allograft and peripheral blood leukocytes with peak levels detected 7 days following liver transplantation. Interestingly, AIF-1 expression increased 2-fold in acutely rejecting liver allografts compared to chronically accepted livers on days 5, 7 and 10 after transplantation. AIF-1 expression in peripheral blood leukocytes was also significantly greater in the rejection model than in the acceptance model. Flow cytometric analysis of peripheral blood leukocytes demonstrated that AIF-1 was expressed in ED2-positive cells, a marker for Kupffer cells. In vitro studies showed that AIF-1 expression in Kupffer cells was up-regulated by coculture with Th1 cytokines. However, neither LPS nor Escherichia coli (E. coli) administration had an affect on AIF-1 expression. These data indicate that high levels of AIF-1 expression reflect aggressive liver allograft rejection and suggest a role for monitoring AIF-1 in peripheral blood leukocytes as a monitor for increased immunosuppression.     

Acute Pancreas Allograft Rejection Is Associated With Increased Risk of Graft Failure in Pancreas Transplantation

The effect of acute allograft rejection (AR) on long‐term pancreas allograft function is unclear. We retrospectively studied 227 consecutive pancreas transplants performed at our institution between January 1, 998 and December 31, 2009 including: 56 simultaneous pancreas and kidney (SPK), 69 pancreas transplantation alone (PTA); and 102 pancreas after kidney (PAK) transplants. With a median follow‐up of 6.1 (IQR 3–9) years, 57 patients developed 79 episodes of AR, and 19 experienced more than one episode. The cumulative incidence for AR was 14.7%, 19.7%, 26.6% and 29.1% at 1, 2, 5 and 10 years. PTA transplant (hazards ratio [HR] = 2.28, p = 0.001) and donor age (per 10 years) (HR = 1.34, p = 0.006) were associated with higher risk for AR. The first AR episode after 3 months post PT was associated with increased risk for complete loss (CL) (HR 3.79, p < 0.001), and the first AR episode occurring during 3‐ to 12‐month and 12‐ to 24‐month periods after PT were associated with significantly increased risk for at least partial loss (PL) (HR 2.84, p = 0.014; and HR 6.25, p < 0.001, respectively). We conclude that AR is associated with increased risk for CL and at least PL. The time that the first AR is observed may influence subsequent graft failure.