The effect of cyclosporin on endothelin levels after orthotopic liver transplantation in rats

To assess the effects of cyclosporin (CyA) on endothelin-1 (ET-1) in rat liver allograft rejection, we evaluated ET-1 expression in samples obtained from BN(RT1ⁿ)-to-BN (group 1) rats, DA(RT1^a)-to-BN (group 2) rats, and DA-to-BN rats treated with 5 mg/kg per day of CyA (group 3). Serum and hepatic ET-1 levels, determined by a radioimmunoassay, remained unchanged in group 1. In group 2, the ET-1 levels peaked on postoperative day (POD) 5 in the liver at 344 ± 31.6 pg/g wet, and on POD 7 in the serum at 38.7 ± 13.1 pg/ml. In group 3, hepatic and renal ET-1 levels showed a progressive increase until POD 10, while serum ET-1 levels remained unchanged. In conclusion, acute rejection caused a temporary increase in the ET-1 level in both the serum and the liver in the early postoperative period what might have been caused by endothelial damage due to ongoing, acute rejection. CyA caused a time-dependent increase in the ET-1 level in both the liver and the kidney without an increase in the serum ET-1 level. The serum ET-1 level might have been affected by the clearance of ET from the liver or kidney. Received: 10 April 1996 Received after revision: 29 October 1996 Accepted: 12 November 1996     

Selective treatment of early acute rejection after liver transplantation: Effects on liver,infection rate,and outcome

To evaluate the results of selective treatment of biopsy-proven mild acute rejection episodes, we retrospectively studied 1-week liver biopsies of 103 patients with a primary liver graft in relation to liver function tests. The overall incidence of rejection was 35 %. In four patients the biopsy showed histological features consistent with rejection; in 27 patients it showed mild acute rejection (grade 1), and in 5 patients it showed moderate acute rejection (grade 2). Study group 1 consisted of 19 untreated patients with grade 1 rejection and group 2 of 8 treated patients with grade 1 rejection. At 30 and 90 days, no differences in liver function tests were found. The infection rate, histology after 1 year, and survival in the two groups did not differ. It may, therefore, be concluded that withholding treatment in histologically proven mild acute rejection is possible in selected patients based on histological, biochemical, and clinical criteria. This may reflect the functional diversity of morphologically similar lymphocytic infiltrates observed in graft biopsies showing features of mild acute rejection.Liver Transplant Group     

Selective treatment of early acute rejection after liver transplantation: effects on liver, infection rate, and outcome

Abstract To evaluate the results of selective treatment of biopsy-proven mild acute rejection episodes, we retrospectively studied 1-week liver biopsies of 103 patients with a primary liver graft in relation to liver function tests. The overall incidence of rejection was 35 %. In four patients the biopsy showed histological features consistent with rejection; in 27 patients it showed mild acute rejection (grade 1), and in 5 patients it showed moderate acute rejection (grade 2). Study group 1 consisted of 19 untreated patients with grade 1 rejection and group 2 of 8 treated patients with grade 1 rejection. At 30 and 90 days, no differences in liver function tests were found. The infection rate, histology after 1 year, and survival in the two groups did not differ. It may, therefore, be concluded that withholding treatment in histologically proven mild acute rejection is possible in selected patients based on histological, biochemical, and clinical criteria. This may reflect the functional diversity of morphologically similar lymphocytic infiltrates observed in graft biopsies showing features of mild acute rejection.     

热休克蛋白70与大鼠肝移植术后早期急性排斥反应的相关性研究

目的对热休克蛋白70（heat shock protein70,HSP70）与大鼠肝移植术后早期急性排斥反应进行相关性研究．探讨其中的免疫学机制。方法建立改良“二袖套法”大鼠原位肝移植模型,将大鼠分为A组（冷保存1h同基因组）,B组（冷保存18h同基因组）,C组（冷保存1h异基因组）,D组（冷保存18h异基因组）,术后收集大鼠肝脏组织和血清．用免疫组织化学及Western blotting方法检测HSP70在移植肝脏中的表达,观察其与病理学检查的相关性结果冷保存时间的延长可以诱导移植肝组织HSP70的高表达,移植肝HSP70表达水平的升高与大鼠原位肝移植术后早期急性排斥病理学评分之间存在着明显的正相关性（P〈0．05,r=0.928）．结论HSP70的升高可能与大鼠原位肝移植术后急性排斥的早期发生以及大鼠移植术后2周存活率密切相关．     

趋化因子IP-10及其受体CXCR3在大鼠肝移植急性排斥反应中的作用

目的探讨趋化因子IP-10及其受体CXCR3的表达在大鼠肝移植手术前后的动态变化,分析其与肝移植急性排斥(acute rejection,AR)的关系。方法改良"二袖套"法建立大鼠原位肝移植模型和急性排斥模型,分为4组:手术创伤组,肝移植无排斥组,肝移植急性排斥组,FK506组。ELISA法检测血清IP-10表达。流式细胞仪检测外周血淋巴细胞CXCR3的表达,用Cellquest软件分析阳性细胞百分率。半定量RT-PCR检测各组第7天肝脏组织CXCR3-mRNA的表达。结果①各组大鼠手术后外周血趋化因子IP-10及其受体CXCR3表达明显升高;②AR组在术后第5天起IP-10及其受体CXCR3表达均高于各对照组(P0.01);③AR组大鼠在移植后第7天,外周血IP-10和CXCR3的表达随着AR组RAI积分的升高而逐步升高,相关系数分别为0.89和0.92(P0.05)。结论血清中趋化因子IP-10及其受体CXCR3的高表达与AR密切相关,有望成为诊断AR较特异、敏感的指标。     

The impact of late acute rejection after cadaveric kidney transplantation

BACKGROUND: Acute graft rejection (AR) following renal transplantation results in reduced graft survival. However, there is uncertainty regarding the definition, aetiology and long-term graft and patient outcome of AR occurring late in the post-transplant period. AIM: To determine if rejection episodes can be classified by time from transplantation by their impact on graft survival into early acute rejection (EAR) and late acute rejection (LAR). MATERIALS AND METHODS: 687 consecutive adult renal transplant recipients who received their first cadaveric renal transplant at a single centre. All received cyclosporine (CyA)-based immunosuppression, from 1984 to 1996, with a median follow-up of 6.9 yr. Details were abstracted from clinical records, with emphasis on age, sex, co-morbid conditions, HLA matching, rejection episodes, patient and graft survival. ANALYSIS: Patients were classified by the presence and time to AR from the date of transplantation. Using those patients who had no AR (NAR) as a baseline, we determined the relative risk of graft failure by time to rejection. The characteristics of patients who had no rejection, EAR and LAR were compared. RESULTS: Compared with NAR, the risk of graft failure was higher for those patients who suffered a rejection episode. A much higher risk of graft failure was seen when the first rejection episode occurred after 90 d. Thus, a period of 90 d was taken to separate EAR and LAR (relative risk of 3.06 and 5.27 compared with NAR as baseline, p<0.001). Seventy-eight patients (11.4%) had LAR, 271 (39.4%) had EAR and 338 (49.2%) had NAR. The mean age for each of these groups differed (LAR 39.6 yr, EAR 40.8 yr compared with NAR 44 yr, p<0.003). The 5-yr graft survival for those who had LAR was 45% and 10-yr survival was 28%. HLA mismatches were more frequent in those with EAR vs. NAR (zero mismatches in HLA-A: 36 vs. 24%, HLA-B: 35 vs. 23% and HLA-DR: 63 vs. 41%, p<0.003). There was no difference in mismatching frequency between NAR and LAR. CONCLUSIONS: AR had a deleterious impact on graft survival, particularly if occurring after 90 d. AR episodes should therefore be divided into early and late phases. In view of the very poor graft survival associated with LAR, it is important to gain further insight into the main aetiological factors. Those such as suboptimal CyA blood levels and non-compliance with medication should be further investigated with the aim of developing more effective immunosuppressive regimens in order to reduce the incidence of LAR.     

肝移植急性排斥反应的诊断与治疗

目的 总结肝移植术后急性排斥反应(AR)的诊治经验。方法 回顾性分析57例肝移植患者术后AR的发生率和治疗结果。结果 21例患者术后因肝功能异常而行38次移植肝活检(术后6～91d)，11例15次发生AR，其中8例为单次，3例为两次或两次以上，轻度11次，中重度4次，AR发生率为19．3％(11／57)。20次为单纯保存-再灌注损伤(PRI)。3次为药物中毒。所有AR均经激素冲击或调整FK506剂量后缓解。结论 移植肝活检对于AR的诊断与鉴别诊断有重要意义，只要给予及时的诊断与治疗，AR一般是可逆的。     

血管内皮生长因子在大鼠肝移植急性排斥反应时的表达

目的 研究血管内皮生长因子(vascular endothelial growth factor,VEGF)在大鼠肝移植急性排斥反应(acute rejection reaction,ARR)中的表达水平,探讨VEGF在细胞介导的肝移植ARR时免疫炎症反应和血管新生的关键中介分子.方法 采用ELISA法及免疫组化EnVision法对VEGF在大鼠肝移植ARR时的表达及血浆中的水平进行了检测.结果 ELISA血清及免疫组化检测显示VEGF在急性排斥组被检测到的水平和表达均比在对照组明显增高,差异有统计学意义(P＜0.05),且以术后2天水平最高.结论 VEGF在移植排斥反应中起着重要作用,其表达水平和移植物存活时间有非常密切关系,并且它是一种早期就表达的作用因子.     

Histologic eosinophilia as an aid to diagnose acute cellular rejection after living donor liver transplantation

The significance of histologic eosinophilia in the diagnosis of acute cellular rejection (ACR) after living donor liver transplantation was evaluated. A retrospective analysis was performed on 185 liver biopsy specimens to determine the presence of eosinophil infiltration around the portal tracts. Data were collected and analyzed to determine whether there was a correlation between ACR and the maximum eosinophil counts per portal triad (Em) and the rate of portal triads that included at least one eosinophil (Er). A receiver operating characteristic curve revealed the best cut-off value of Em and Er as 2% and 8% respectively. The sensitivity and specificity of an Em of two to predict ACR were 54% and 84% respectively. The sensitivity and specificity of Er were 72% and 65% respectively. One-way analysis of variance revealed that both Em and Er correlated with ACR severity. Histologic eosinophilia can be a useful parameter for confirming the occurrence of ACR and for evaluating ACR severity.     

Severe late acute allograft rejection in a child after living-related auxiliary partial orthotopic liver transplantation for ornithine transcarbamylase deficiency

Auxiliary liver transplantation (ALT) is known to correct liver-based metabolic disorders. However, it remains unclear whether the presence of a native liver influences the long-term prognosis of ALT for metabolic diseases. We reported on a 4-yr-old girl who had undergone living-related auxiliary partial orthotopic liver transplantation (APOLT) for ornithine transcarbamylase deficiency and experienced severe late acute rejection 18 months after liver transplantation, during weaning of immunosuppressive agents. Results of histological analysis of the graft indicated very severe acute rejection (rejection activity index, 9/9), and computed tomography revealed graft liver atrophy. These observations suggest the possibility that severe rejection might occur in APOLT, especially during weaning of immunosuppression.     

再次背驮式肝移植并肾移植治疗肝移植术后慢性排斥并肾功能不全(附一例报告)

目的 探讨用原位背驮式肝移植并肾移植术治疗先天性多囊病肝移植术后慢性排斥并肾功能不全。方法 供肝取自同血型脑死亡供体;顺利切除原病肝,行原位再次背驮式新肝植入及右髂窝肾移植。结果 再次背驮式肝移植并肾移植手术成功,供肝,供肾功能良好,术后１８ｄ死于曲霉菌性腹膜炎所致的多器官功能衰竭。结论 背驮式技术适用于再次肝移植;肝移植术后慢性排斥反应并肾功能不全是联合肝肾移植的适应证。     

Serum interleukin-6 levels as an indicator of acute rejection after liver transplantation in cynomologous monkeys

This study was designed to evaluate whether the sequential monitoring of serum interleukin-6 levels (SIL-6) could be helpful for diagnosing the occurrence of hepatic allograft rejection. An SIL-6 post-transplant study was conducted on nine cynomolgus monkeys which had undergone orthotopic hepatic allotransplantation, six of which were treated with FK-506 (a new immunosuppressant agent isolated fromStreptomyces tsukubaensis) and three of which were not. All the nontreated animals showed biochemical abnormalities from days 5–6, characterized by a marked elevation of serum alkaline phosphatase levels, and they eventually died on days 8, 12, and 63 (group I). Acute cellular rejection was confirmed by histological study of the hepatic grafts taken at autopsy or biopsy. On the other hand, four of the treated animals (group IIa) survived more than 30 days. Biochemical examination of this group showed no abnormal signs apart from a slight elevation of alkaline phosphatase (<2000 IU/l). Histological examination carried out around 30 days after transplantation revealed a transient infiltration of polynuclear cells into Glisson's area, with the portal vein and bile duct remaining intact. The remaining two animals (group IIb) died of dehydration and arterial thrombosis on days 5 and 7, respectively. A kinetic study of SIL-6 conducted during the first 2 weeks showed quite different patterns among the three groups. All recipients in group I demonstrated two peaks following grafting on days 1 and 3 or 4, the second peak of above 2.0 U/ml preceding biochemical abnormalities by 2 to 3 days. Conversely, the recipients in group IIa showed a single peak of SIL-6 on day 1, followed by a decrease to less than 2.0 U/ml. Group IIb characteristically showed a high level of SIL-6 on day 1, after which the value remained over 2.0 U/ml. In conclusion, the monitoring of SIL-6 proved useful for detecting the acute rejection of hepatic allografts, the recipients with rejected grafts displaying two peaks of SIL-6 after grafting. The first increment was observed on day 1, which might reflect the stress of the surgical procedure, while the second transient increment of SIL-6, observed on day 3 or 4, might suggest acute hepatic rejection, preceding biochemical abnormalities by 2 to 3 days.     

Incidence,risk factors,and outcome of antithymocyte globulin treatment of steroid-resistant rejection after liver transplantation

We retrospectively analyzed the incidence and outcome of steroid-resistant rejection (SRR) during the first 6 months after OLT in 126 patients receiving triple immunosuppression. A total of 95 patients either did not experience acute rejection at all or had acute rejection that subsided without additional treatment. A total of 31 patients had biopsy-proven acute rejection that required therapy: 18 patients had acute rejection that responded to steroid therapy (steroidsensitive rejection, SSR); the remaining 13 patients had SRR and received ATG. At the onset of acute rejection, no differences in clinical, biochemical, or immunological parameters were present between patients with SSR and SRR. However, the histological grade of acute rejection in the initial biopsy was higher in patients with SRR (P=0.05). ATG treatment was effective in 10 of the 13 patients and was not associated with an increased incidence of opportunistic infections. Patient and graft survival rates at 2 years were comparable in the three groups. These data show that the incidence of SRR during the first 6 months after OLT is low, and that its treatment with ATG is both effective and well tolerated.     

血浆置换治疗肾移植术后抗体介导的急性排斥反应的疗效观察

目的 探讨血浆置换治疗肾移植术后抗体介导的急性排斥反应的效果. 方法 2011年1月至2013年9月行同种异体肾移植术后发生抗体介导的急性排斥反应患者5例,男2例,女3例.年龄41252岁,平均46岁.术前诊断均为慢性肾功能不全尿毒症期,行规律血液透析.术后采用环孢素[5 mg/(kg·d)]或他克莫司[0.1 mg/(kg·d)],以及吗替麦考酚酯(1.5 g/d)和糖皮质激素行免疫抑制治疗.术后2周内均经移植肾穿刺病理检查及血清供者特异性抗体测定诊断为抗体介导的急性排斥反应.予甲泼尼龙(1 000 mg/d)和抗淋巴细胞球蛋白(250 mg/d)治疗无效,在环孢素[5 mg/(kg·d)]或他克莫司[0.1 mg/(kg·d)],以及吗替麦考酚酯(1.5 g/d)和糖皮质激素免疫抑制治疗的基础上,5例患者均分别行血浆置换7次.4例原发病为慢性肾小球肾炎,术前血清肌酐为(784±154) μmol/L,术后2周内开始进行血浆置换;1例原发病为抗肾小球基底膜肾病,术前血清肌酐水平为935 μmol/L,术后35 d开始进行血浆置换. 结果 4例原发病为慢性肾小球肾炎患者分别经7次血浆置换治疗后排斥反应得到逆转,肾功能恢复良好,随访3个月时血清肌酐水平为(113±12) μmol/L.原发病为抗肾小球基底膜肾病患者,血浆置换后排斥反应未得到纠正,移植肾功能未恢复,随访3个月时血清肌酐水平524 μmol/L,继续血液透析治疗,随访12个月时血清肌酐水平758 μmol/L,超声检查示移植肾萎缩,予口服他克莫司0.5mg/d治疗. 结论 2周内应用血浆置换能有效地逆转肾移植术后患者抗体介导的急性排斥反应.     

Comprehensive immunohistological analysis of the endothelin system in human kidney grafts.

BACKGROUND: In experimental models of renal transplantation, upregulation of the endothelin (ET) system and amelioration of renal injury by ET-receptor blockers have been documented. In contrast, little information is available on the expression of the ET system in human kidney allografts. It was the purpose of the present study to analyse by immunohistology the expression of ET-1 as well as of the two ET receptors (ET-RA and ET-RB) in the different cells and compartments of kidney grafts and control kidneys. METHODS: Fifty-five graft biopsies were taken from 55 kidney allograft recipients (mean age: 32+/-2.8 years) who were all on a calcineurin inhibitor. The indication for biopsies was delayed graft function or suspected rejection. The underlying diagnoses were acute allograft rejection (n = 14), chronic allograft nephropathy (n = 14), cyclosporin A (CSA) toxicity (n = 10), post-operative acute tubular necrosis (ATN) (n = 11) and recurrent primary disease (n = 6). As control, tissues of non-grafted kidneys with ATN (mean age: 35+/-24 years), of primary glomerulonephritis (mean age: 69+/-10 years) and of non-tumour-bearing parts of eight tumour nephrectomy specimens (mean age: 67+/-5 years) were assessed. The biopsies were scored using the 1997 Banff criteria. Expression of ET-1, ET-RA and ET-RB as well as of vascular endothelial growth factor was evaluated by immunohistochemistry and a semi-quantitative scoring system. Interstitial infiltrating cells were characterized using antibodies against T cells, B cells and macrophages (CD3, CD20 and CD68). RESULTS: Control cases showed only faint expression of ET-1 in glomeruli (in podocytes and endothelial cells), whereas marked expression was seen in distal, but less in proximal tubular cells. The interstitium was completely negative. ET-1 expression was seen in vascular endothelial cells (VEC) and vascular smooth muscle cells (VSMC). Only faint expression of ET-RA and ET-RB was found in glomeruli and tubuli (distal more than proximal). Marked ET-RA and ET-RB expression was seen in VEC and VSMC. In all transplanted kidneys, irrespective of the underlying diagnosis, expression of ET-1, ET-RA and ET-RB was markedly higher compared with control kidneys. ET-1 was strikingly upregulated in glomeruli and tubuli, but surprisingly not in the vasculature of grafts with CSA toxicity. Expression of ET-RB was markedly increased in CSA toxicity in glomeruli, tubuli and vessels. In grafts with ATN and acute rejection, pronounced expression of ET-RA was noted. There was a strong correlation between proteinuria and expression of ET-1 in glomeruli and proximal tubuli and of ET-RB in proximal tubuli. CONCLUSIONS: The above data in human kidney allograft biopsies are consistent with an important role of the ET system in different types of renal allograft damage. This finding extends and clarifies the somewhat contradictory results in animal models.     

大黄素对大鼠移植肝细胞凋亡的作用

目的探讨大黄素对大鼠肝脏移植术后肝细胞凋亡的作用。方法建立大鼠肝移植模型,分为三组:对照组,供受体均为LEW大鼠;移植组;供体为LEW大鼠,受体为BN大鼠;大黄素组,在移植组基础上,移植术后每日以大黄素50mg·kg-1腹腔注射。分别于术后第1,3,5,7天各取6只移植大鼠的肝脏,TUNEL法染色,检测肝脏细胞凋亡。以肝脏细胞凋亡阳性细胞数占总肝脏细胞数的百分比作为肝细胞凋亡指数(AI)。结果移植组肝细胞于术后第1天即已出现凋亡,第3天明显增加,第7天达到高峰。大黄素组各时间点肝细胞凋亡指数明显小于对应的移植组(P<0.01)。结论大黄素对肝移植急性排斥反应中肝细胞凋亡有显著抑制作用。     

The long-term impact of liver transplantation on kidney function in familial amyloidotic polyneuropathy patients

The aim of the study is to evaluate the long-term kidney function after liver transplantation (LTx) in familial amyloidotic polyneuropathy (FAP) Portuguese type patients and compare the findings with patients transplanted for chronic liver disease of other origin. We analysed the medical records of 32 FAP patients who underwent transplantation between 1990 and 1999 with a follow-up of more than 1 year after LTx. The control group consisted of 61 patients who had undergone LTx for chronic liver disease. Kidney function was measured by the glomerular filtration rate (GFR), serum creatinine and urea. There were no differences between the groups in creatinine and urea levels during the follow-up. However, during the first year after transplantation, the increase in creatinine and urea was significantly higher in the control group (P < 0.01). The decline in GFR after transplantation was also more pronounced in the controls (P < 0.01). Initially after LTx, kidney function deteriorated in both FAP and control patients, but the deterioration was more pronounced in the controls. The decline of the FAP patients' kidney function after LTx was not more pronounced than that observed in control patients, although many FAP patients' kidney function was impaired before the procedure, suggesting that LTx may halt the progression of kidney damage caused by amyloid deposition.