Gemcitabine does not prevent acute rejection of the transplanted liver in rats

Our study was designed to determine effect of gemcitabine on acute rejection of liver in rats. Liver transplantation was performed in rats of the Dark Agouti (DA) and Lewis (LEW) strains. Recipients were divided into three groups: A, DA-to-LEW without immunosuppression; B, DA-to-LEW, treated with cyclosporine A; C, DA-to-LEW, treated with gemcitabine. Immunosuppressants were subcutaneously injected for seven consecutive days after transplantation. On day 7, blood samples and liver graft tissue specimens were harvested. Group A showed severe rejection changes (RAI 8/9); in group B no rejection changes were present (RAI 0/9), and in group C moderate rejection changes were observed (RAI 6/9). Differences were significant between B vs C and A vs C groups; P<0.05. Serum creatinine and urea levels in the gemcitabine group were significantly lower than those in the cyclosporine A group. We did not confirm gemcitabine ability to prevent liver allograft rejection.     

不同品系大鼠之间肝移植排斥反应的比较研究

目的比较不同品系大鼠之间肝移植排斥反应特点。方法依据大鼠不同品系分成对照组SD→SD(A组),实验组Wistar→SD(B组)、Lewis→BN(C组)以及DA→Lewis(D组)4个组,采用Kamada双袖套法建立大鼠原位肝移植模型。比较各组受体大鼠术后10 d血清肝功能指标[丙氨酸转氨酶(ALT)、总胆红素(TB)和白蛋白(Alb)]、血清白细胞介素(IL)-2和IL-10水平、急性排斥反应组织病理学分级和术后平均生存时间。结果与A、B组比较,C组和D组的血清ALT和TB明显升高,Alb明显降低,差异均有统计学意义(均为P0.05)。与C组比较,D组的TB明显升高,Alb明显降低,差异均有统计学意义(均为P0.05)。与A组比较,B、C、D组大鼠血清IL-2和IL-10水平均明显升高,C组和D组的IL-2/IL-10亦明显升高,差异有统计学意义(均为P0.05)。与B组比较,C组和D组大鼠血清IL-2水平明显升高,D组的IL-2/IL-10亦明显升高,差异有统计学意义(均为P0.05)。与C组比较,D组大鼠血清IL-2水平明显升高,D组的IL-2/IL-10亦明显升高,差异有统计学意义(均为P0.05)。肝组织病理学检查显示,A组大鼠肝组织未见明显排斥反应,排斥活动性指数(RAI)评分(1.8±0.7)分;B组RAI评分(3.1±1.3)分,属轻度或不明确性排斥反应;C组RAI评分(6.9±0.8)分,属中~重度排斥反应;D组RAI评分(8.8±0.5)分,属重度排斥反应。各组间RAI评分比较以及组间两两比较,差异均有统计学意义(均为P0.05)。A组、B组、C组、D组受体大鼠术后平均生存时间分别为(119.3±1.9)d、(116.9±8.3)d、(53.4±6.1)d、(12.1±2.4)d,A组与B组的生存时间比较差异无统计学意义,余各组两两比较差异均有统计学意义(均为P0.05)。结论 4种组合中,大鼠DA→Lewis模型排斥反应最剧烈,Lewis→BN次之,而Wistar→SD最轻,接近于免疫耐受。     

大鼠原位肝移植术后急性排斥反应中 Fractalkine的表达及意义

目的利用直视下套入式吻合肝动脉加袖套法吻合门静脉建立的大鼠全血供肝移植模型,研究同种异体大鼠肝移植术后早期急性排斥反应中Fractalkine（Fkn）的表达情况。方法制备SD-Wistar大鼠全血供肝移植模型,随机分为两组,每组20只。对照组：于移植术后第1天开始腹腔内注射生理盐水（3ml/kg）;实验组：于移植术后第1天开始腹腔内注射环孢素A（3mg/kg）。术后观察大鼠一般情况,并于第3、5及7天分别处死5只大鼠,取肝脏标本,观察移植肝组织排斥反应强度（rejection activity index,RAI）及Fkn表达情况;余大鼠观察生存时间。结果对照组存活18只,实验组存活19只。实验组较对照组术后一般情况好,存活时间为19.50±4.51d,与对照组7.60±1.60d比较,差异有统计学意义（P〈0.05）。组织学观察：对照组移植肝小叶结构清楚,汇管区增大,大量淋巴细胞浸润;实验组移植肝组织RAI明显降低。术后第3、5及7天,对照组RAI为3.80±0.35、5.90±0.87及7.50±1.30,实验组为3.10±0.21、3.90±0.41及4.50±0.52。两组术后第7天RAI比较差异有统计学意义（P〈0.01）。免疫组织化学观察：两组移植肝组织中均见细胞浆或细胞膜有棕色颗粒的Fkn表达,但实验组表达较弱。术后第3、5及7天,对照组Fkn细胞数为8.20±0.57、21.30±3.30及25.70±4.91,实验组分别为8.30±0.56、10.30±0.67及11.70±1.23;两组第5、7天比较差异有统计学意义（P〈0.01）。结论Fkn参与了同种异体大鼠肝移植急性排斥反应,可作为诊断大鼠肝移植急性排斥反应发生的指标之一。环孢素A可以通过抑制Fkn表达来抑制排斥反应发生的强度。     

Prevention of lung allograft rejection by combined treatment with adhesion molecule antibodies and cyclosporine

Infiltration of leukocytes into the lung allograft is regulated by adhesion molecules during acute rejection. The purpose of this study was to assess the effect of monoclonal antibodies against ICAM-1 (1A29) to prevent rejection after lung transplantation. Left lateral orthotopic lung transplantation was performed using Dark Agouti rats as donors and Lewis rats as recipients. Recipients received 1A29 alone (group A), cyclosporine A alone (group B), a combination of both drugs (group C) or no therapy (group D). Animals were killed on day 5 and 10, respectively. Rejection was graded by histology. Myeloperoxidase activity (MPO) was measured in the graft. In allografts treated with cyclosporine and 1A29 histologically a lower grade of rejection was seen and less MPO were detected compared to groups A, B and D. Anti-ICAM-1 monoclonal antibodies alone as well as a subtherapeutic dose of cyclosporine are not effective to prevent acute allograft rejection after lung transplantation. However, the combination of both strategies significantly reduces rejection in this model.     

芳香烃受体的激活在大鼠肝移植免疫应答中的作用及机制研究

目的芳香烃受体（AhR）是一种配体激活性转录因子，可以影响T细胞分化，在多种免疫性疾病中发挥作用，本研究主要探索AhR在大鼠肝移植免疫应答中的作用以及机制。 方法建立大鼠肝移植同基因耐受模型和异基因排斥模型，术后1、3、7、14 d检测各组AhR和程序性死亡（PD-1）表达量。对肝移植排斥模型进行不同处理并分组。A组：对照组，腹腔注射（i.p.）羧甲基纤维素钠（CMC-Na） 1 ml；B组：200 mg·kg^-1·d^-1的3,4-DAA（i.p.）；C组：10 mg·kg^-1·d^-1的CH223191（i.p.）；D组：200 mg·kg^-1·d^-1的3,4-DAA和10 mg·kg^-1·d^-1的CH223191（i.p.）。7 d后采集受体外周血检测肝功能指标及CD4⁺Foxp3⁺T细胞和CD4⁺PD-1⁺T细胞占比，收集大鼠移植肝组织进行病理学检查，采用Banff分级评估排斥活动指数（RAI），并通过免疫印迹法、qPCR检测移植肝组织AhR、PD-1的变化。 结果在肝移植排斥模型中AhR和PD-1表达量随着时间逐渐降低，在肝移植耐受模型中逐渐增加。与A组相比，B组受体大鼠RAI降低，存活时间延长，移植排斥减轻；而C组肝功能指标明显上调，RAI升高，移植排斥反应加重；D组与A组差异不明显。AhR和PD-1表达以及CD4⁺Foxp3⁺T细胞、CD4⁺PD-1⁺T细胞占比在受体中呈现相同的趋势。 结论随着肝移植排斥反应的加重，AhR和PD-1表达降低，反之亦然。AhR激活可通过增加调节性T细胞的比例和PD-1的表达，有效降低大鼠肝移植排斥反应的发生。AhR有可能成为临床肝移植术后排斥治疗的新靶点。     

供体骨髓间充质干细胞抑制肝移植大鼠免疫排斥的研究

目的 探讨供体骨髓间充质干细胞(MSCs)抑制肝移植大鼠免疫排斥的作用.方法 实验按照随机数字表法分为3组,每组14对雌性大鼠,建立Wistar-SD大鼠原位肝移植模型.A组:肝移植术中输注生理盐水;B组:肝移植术后隔天一次给予他克莫司(0.25 mg/kg)灌胃2周;C组:肝移植术中输注与A组同剂量的雄性Wistar大鼠的MSCs.观察术后第10天肝功能的变化、病理改变、TGF-β1与IL-10的表达、Y染色体定位及受体存活时间.采用完全随机化设计多组比较方差分析AST、ALT值,LSD法分析组间差异;病理分级采用ridit法分析;Kaplan-Meier法计算存活率,Log-rank检验进行生存分析.结果 肝移植术后第10天A、B、C组ALT分别为(756±104)、(197±49)、(103±31)U/L,AST分别为(635±134)、(331±78)、(150±38)U/L,3组ALT和AST比较差异均有统计学意义(F=158,265,P<0.05);两两比较发现,B组及C组均优于A组,且C组优于B组.A组ridit均值为0.8333,为重度排斥,B、C两组ridit均值分别为0.4583、0.2083,排斥反应较A组显著减轻,且C组较B组级别更低.A、B、C组TGF-β1阳性表达率分别为18%±5%、69%±20%及85%±24%,3组比较差异有统计学意义(F=191,P<0.01).A、B、C组IL-10阳性表达率分别为21%±5%、75%±14%及91%±21%,3组比较差异有统计学意义(F=672,P<0.01);B、C两组TGF-β1和IL-10呈强阳性表达且C组比B组更明显,而A组则为弱阳性表达.原位杂交检测发现,C组含有带Y染色体的雄性MSCs,主要在汇管区聚集.A、B、C组大鼠50 d存活率分别为0、10%、90%,3组大鼠存活率比较差异有统计学意义(χ~2=36,P<0.01),C组中位存活时间为63 d,较A组11 d延长,且C组长于B组.结论 肝移植同时向供体输注MSCs能够抑制受体对移植肝的免疫排斥.     

猪肝肠联合移植免疫耐受的实验研究

目的 研究猪肝肠联合移植中肝移植物对同源小肠移植物免疫耐受的作用.方法 70头杂交长白猪分为4组,A、B、C组为辅助性同种异体肝肠联合移植(每组20头);D组为节段性间种异体小肠移植(10头).移植后A、D组未用免疫抑制剂治疗,B、C组分别采用常规剂量和小剂量的环孢素和甲基强的松龙治疗.结果 A组术后小肠移植物较D组排斥反应时间延迟,程度明显减轻(P＜0.05).常规剂量的B组与小剂量的C组在术后存活时间、排斥反应开始时间以及排斥反应程度方面差异无统计学意义(P＞0.05).结论 猪同种异体肝肠联合移植中肝移植物可以诱导同源小肠移植物免疫耐受.     

Immunosuppression without prednisone after liver transplantion is safe and associated with normal early graft function: preliminary results of a randomized study

Abstract Prednisone has been commonly considered the mainstay of immunosuppressive therapy after liver transplantation. Recent data suggest that prednisone withdrawal late after transplant reduces complications without affecting graft function. We report here the preliminary results of an open-label, randomized study aimed at investigating whether prednisone therapy can be completely avoided during the first 3 months after transplantation. Twenty-seven consecutive patients were randomized to receive double (group A: cyclosporine and azathioprine) or triple (group B: prednisone, cyclosporine, and azathioprine) immunosuppressive therapy after liver transplantation. Six patients died within the first 3 weeks in each group and were excluded from the calculations. The present results refer to 10 patients in group A and 11 in group B. The actuarial 1-year survival did not differ between the two groups (90.9 % vs 88.8 %). There were no differences with respect to infectious complications or episodes of histological acute graft rejections. Only one severe acute rejection occurred in group A and two in group B. During the first month after transplant, liver and kidney functions tended to be better in the group of patients treated without prednisone, although there were no differences in the mean cyclosporine blood levels. These data, though preliminary, indicate that early immunosuppression without the use of prednisone is safe and tends to be associated with improved liver and renal functions compared to conventional triple therapy.     

血清肝炎病毒标志物阳性肾移植患者临床用药特点

目的 探讨血清肝炎病毒标志物阳性。肾移植患者术后临床用药特点。方法 40例同种异体。肾移植患者，男22例，女18例。年龄30～56岁。其中乙型肝炎感染29例、丙型肝炎感染9例、乙型肝炎合并丙型肝炎感染2例。患者肝功能正常，随机分为普乐可复组（n=20），环孢素A组（n=20）。观察患者术后肝、肾功能情况及人／。肾存活率。结果 40例患者术后随访2年，普乐可复组肝功能异常发生率、急性排斥反应发生率明显低于环孢素A组（分别为15％vs30％，5％vs20％），2组2年人／肾存活率均为100％。结论 血清肝炎病毒标志物阳性患者接受肾移植术后首选普乐可复作为基础免疫制剂方案，可减少排斥反应发生率，对肝脏的损害程度轻。     

脾脏在小鼠同种肝移植排斥反应中的作用

目的 观察脾切除在小鼠同种肝移植急性排斥反应过程中的作用.方法 双袖套法建立小鼠原位肝移植模型,随机分为3组,即建模保留脾脏组、建模3 d后切除脾脏与建模同时切除脾脏组,各组于移植术后14 d处死,ELESA法测定血清IgM水平;肝功能检测采用速率法;流式细胞仪检测CD4与CD8T细胞亚群;并同时行肝脏及脾脏的病理形态观察.结果 建模保留脾脏组、建模3 d后切除脾脏与建模同时切除脾脏组血清IgM水平分别为3.0181±0.4627、3.0936±0.4559、3.1953±0.4449,各组间差异无统计学意义(P＞0.05);ALT水平分别为108.6875±20.3657、83.0000±22.7799、76.8000±19.5784,差异有统计学意义(P＜0.05);AST水平分别为:105.3750±29.0583、93.0000±22.7799、93.2000±33.4220,各组间差异无统计学意义(P＞0.05);CD46+/CD8+T细胞分别为:1.9162±0.2778、1.5654±0.4750、1.4616±0.2762,差异有统计学意义(P＞0.05);3组肝脏间质及汇管区淋巴细胞浸润程度依次减弱,供肝灶状坏死程度逐渐减轻,在保留脾脏组中建模后第14天脾脏边缘区及淋巴鞘较建模同时切除的脾脏增宽.结论 在小鼠同种异体肝移植排斥反应中细胞免疫起主要作用,脾切除可部分抑制同种异体肝移植急性排斥反应,保护供体肝脏.     

吲哚胺2,3双加氧酶在诊断肝移植急性排斥中的作用

目的 探讨外周血吲哚胺2,3双加氧酶(IDO)基因表达对大鼠肝移植急性排斥反应(AR)的诊断价值.方法 建立大鼠原位肝移植模型,分为4组:A组:同基因移植组(Wistar-Wistar,n=32);B组:异基因移植组(SD-Wistar,n=32);C组:异基因移植+长期环孢素A组(CsA,n=32);D组:异基因移植+短期CsA组(用药剂量同C组,第3天起停药,n=32).应用实时聚合酶链反应(Real-time PCR)方法 分别检测术后第0、1、2、3、4、5、7、9天外周血IDO mRNA值,同时检测各时间点血清谷草转氨酶(AST)、总胆红素(T-BIL)、碱性磷酸酶(ALP)水平,并取肝脏病理切片.结果 A组外周血IDO mRNA呈持续低水平,AST、T-BIL、ALP逐渐降至正常,病理无排斥反应.C组检测结果 与A组相似.B组IDO mRNA显著上升为术后第2天(P<0.05),AST、T-BIL、ALP值显著上升为术后4d(P<0.01),病理切片判断轻度排斥为术后5d(χ2=4.8,P<0.05).D组IDO mRNA显著上升为术后第4天(P<0.05),AST、T-BIL、ALP值显著上升为术后5d(P<0.01),病理切片判断轻度排斥为术后7d(χ2=4.8,P<0.05).结论 外周血IDO mRNA检测可较病理检查更早诊断大鼠肝移植AR的发生,且方法 简单、安全.

Abstract：

Objective To study the diagnostic value of indoleamine 2, 3-dioxygenase (IDO) gene expression in acute liver rejection in rat orthotopic liver transplantation model. Methods The rat orthotopic liver transplantation models were divided into four groups: group A, isograft transplantation group (Wistar to Wistar); group B, allograft transplantation (SD to Wistar); group C, allograft transplantation and cyclosporine; Group D, allograft transplantation and cyclosporine (the drug was withdrawn on the 3rd day after the operation). The samples (peripheral blood and liver tissue) were obtained on the operation day, 1st, 2nd, 3rd, 4th, 5th, 7th and 9th day post-operation. Luorescent quantitative polymerase chain reaction (PCR), pathological study and serum test were performed on each sample. Results In groups A and C, the IDO mRNA was expressed at a low level, aspartate aminotransferase (AST), total bilirubin (T-BIL) and alkaline phosphatase (ALP) were decreased to the normal level, and the pathological test found no acute rejection (AR). In group B, the IDO mRNA was dramatically increased on the day 2 (P<0.05), AST, T-BIL and ALP rose dramatically on the day 4 (P<0.05), and mild AR was detected on the day 5 (χ2=4.8,P<0.05). In group D, the IDO mRNA was dramatically increased on the day 4 (P<0.05), AST, T-BIL and ALP rose dramatically on the day 5 (P<0.01), and mild AR was detected on the day 7 (χ2=4.8,P<0.05). Conclusion Comparing with pathological study, detecting IDO mRNA of peripheral blood can diagnose AR of rat at earlier stage and the technique is safe and simple.     

The effects of pretransplant cyclosporine therapy on rats grafted with twelve-hour cold-stored livers--with special reference to reperfusion injury

The effect of pretreatment with cyclosporine on liver preservation was studied using a rat liver transplant model. In a preliminary 1-week survival study, 59 liver transplants were performed. In group A, neither donors nor recipients were treated. In group B, the recipients were pretreated by a 3-day course of CsA (10 mg/kg/day p.o.), but the donors were untreated. In group C, the donor rats were pretreated for 3 days with the same doses of CsA as in group B, but the recipients were not treated. The donor livers in each group were stored for 12 hr at 4 degrees C with Eurocollins solution and transplanted to the recipients. The CsA pretreatment to recipients (group B) significantly improved 1-week survival (57.1%, 8/14, P less than 0.01 versus control group A; 0%, 0/14 or group C; 14.3%, 2/14). To study lipid peroxidation and morphology, 72 rat livers were studied in 9 groups. In summary, CsA pretreatment to recipients resulted in suppression of the increase in MDA levels and amelioration of endothelial injury after transplantation. On the other hand, donor pretreatment exerted dual effects on the grafts; it ameliorated endothelial injury after reperfusion, but its hepatotoxic action exacerbated hepatocellular damage during hypothermic storage. Our study suggests that CsA pretreatment, particularly to recipients, is beneficial in liver preservation for hepatic transplantation. The mechanisms are discussed with regard to ischemia/reperfusion injury to hepatic endothelium.     

辅助性同种异体肝肠联合移植术后急性排斥反应的监测

目的评价猪同种异体辅助性肝肠联合移植术后排斥反应的监测方法。方法将50头杂交长白猪分为3组．A、B组各20头各完成10次猪辅助性带胰头及十二指肠的同种异体肝肠联合移植术．其中B组术后予以免疫抑制治疗：C组10头完成交互的同种异体节段性小肠移植术10例。术后1、3、5、7、14、21及30d经移植肠远端造口取小肠黏膜经常规处理后。分别在光镜和电镜下观察并进行排斥反应评分。结果术后A组出现排斥反应的中位时间为8（7～12）d．迟于c组的5（3～5）d（P〈0．05）。术后1周,A组的排斥反应评分为1．11±0．20。低于C组（2．56±0．18,P〈0．05）;但比B组高（O．20±0．13,P〈O．05）。A组移植术后中位存活时问为9（7～25）d,C组为12（7～20）d．而B组术后全部成活超过30d．与以上两组比较,P〈0.05．差异有统计学意义。结论移植术后排斥反应通过肠造口取材进行监测方便有效。     

Clinical results of immunosuppressive triple therapies in living-related renal transplantation--a single center experience

We reviewed the outcome of three methods employed for living-related renal transplantation (RTx) in our institution to assess triple immunosuppressive regimens. Between January 1989 and July 2003, a total of 35 living-related RTxs were performed at our institution. The immunosuppressive regimen given to 16 patients (group A) was cyclosporine (CsA), steroid and azathoprine (AZ) that given to 9 patients (group B) was tacrolimus (TAC), steroid and AZ and that given 9 patients (group C) was TAC, steroid and mycophenolate mofetil (MMF). Graft survival rate, serum creatinine, proteinuria, acute rejection, chronic allograft nephropathy (CAN), cytomegalovirus (CMV) infection and drug-induced nephropathy were investigated. There was no significant difference in graft survival rate among the three groups. Although serum creatinine levels (mg/dl) at 3 months post-transplant were 1.22+/-0.37 in group A, 1.43+/-0.14 in group B, 1.30+/-0.34 in group C, respectively (p<0.05; A vs. B), there was no significant difference at 1 year post-transplant. Frequency of proteinuria in groups A, B and C was 75.0, 50.0, 25.0%, respectively (p<0.05; A vs. C). The incidences of acute rejection and CAN within 1 year post-transplant were, respectively, 56.3% and 43.8% in group A, 37.5% and 37.5% in group B; and, 25.0% and 12.5% in group C (NS). The incidence of drug-induced nephrotoxicity was 12.5, 50.0% and 37.5% in groups A, B and C, respectively (p<0.05; A vs. B). The triple immunosuppressive therapy including calcineurin inhibitors, especially the regime of TAC, MMF, and steroids decreased the frequencies of proteinuria and rejections, which deteriorated the long-term outcome in living-related RTxs.     

Combination use of suboptimal dose of FK 506 and cyclosporine in canine lung transplantation.

The immunosuppressive potency and the side effects of combination therapy with FK 506 and cyclosporine A were studied in dogs that had undergone lung transplantation. The animals were divided into four groups: group A (one third optimal FK 506 dose: FK 506, 0.03 mg/kg intramuscularly) (n = 5), group B (one third optimal cyclosporine dose: cyclosporine 6 mg/kg orally) (n = 5), group C (one third FK 506 and one third cyclosporine optimal doses): FK 506, 0.03 mg/kg intramuscularly plus cyclosporine 6 mg/kg orally) (n = 5), and group D (half FK 506 plus half cyclosporine optimal doses: FK 506, 0.05 mg/kg intramuscularly, plus cyclosporine, 10 mg/kg orally) (n = 10). Assessments including chest x-ray film, fiberoptic bronchoscopy, hematologic and biochemical tests, FK 506 and cyclosporine blood trough level measurement, right pulmonary artery occlusion test, and histopathologic observations were performed. In group A two of five dogs survived 28 days and three died on postoperative days 7, 14, and 21. In group B one dog survived 28 days and four died on postoperative days 9 (two dogs), 14, and 21. Histologic examination showed severe rejection in both group A and group B. In group C all five dogs survived 28 days but showed mild rejection. In group D one dog died of intestinal bleeding on postoperative day 7 and nine survived 28 days. No pathologic changes were observed except in one case of mild rejection. The ventilation function of the transplanted lung was poor in groups A, B, and C but good in group D. No abnormal rise of FK 506 and cyclosporine trough levels was observed. There were no significant side effects and abnormal hematologic and biochemical data except in one dog in group D. We concluded (1) the combination of FK 506, 0.03 mg/kg, and cyclosporine, 6 mg/kg, is much more effective than either drug used singly, (2) the combination of FK 506, 0.05 mg/kg, and cyclosporine, 10 mg/kg, prevents rejection with tolerable side effects, and (3) no worse side effects are caused by combination therapy with FK 506 and cyclosporine than by either one used singly.     

应用肝移植供体脾淋巴细胞预防受体鼠肝癌复发的研究

目的 探讨肝癌肝移植后应用活化供体脾脏来源的淋巴细胞,对受体实施过继性免疫治疗以预防肝癌复发的可行性.方法 以培养黏附法体外分离培养Wistar大鼠骨髓树突状细胞(DCs)前体细胞,经诱导后与Wistar大鼠肝癌细胞CBRH-7919裂解物抗原共同孵育,形成负载癌抗原的DCs.以此DCs分别诱导受体(Wistar大鼠)、供体(SD大鼠)来源的脾淋巴细胞作为两个实验组C(受体活化组)、D(供体活化组),以未经抗原诱导的受、供体脾淋巴细胞作为对照组A(受体对照组)、B(供体对照组).对比活化、非活化以及供体来源、受体来源的脾淋巴细胞在体外对受体肿瘤细胞的杀伤活性.以上述不同来源和处理的脾淋巴细胞对SD→Wistar大鼠肝移植后肿瘤复发模型行过继性免疫治疗,以盐水治疗作为对照组E,每组6只大鼠,观察各组免疫治疗对于肿瘤浸润淋巴细胞、受体肝脏成瘤率和供肝的排斥反应的影响.结果 DCs诱导脾细胞过程中,培养上清液γ干扰素(IFN-γ)分泌C、D组较A、B组有明显的升高.C、D组对肝癌细胞的杀伤活性较A、B组显著增强,D组较C组显著增强.D组脾细胞回输体内后,在移植后大鼠体内观察到了肿瘤浸润淋巴细胞增多,肿瘤组织坏死和成瘤率下降.免疫治疗前后供肝未见严重排斥反应发生.结论 活化的供体脾细胞较受体脾细胞有更强的肿瘤杀伤效果.应用供体脾淋巴细胞对肝移植受体进行过继性免疫治疗可以在不增加对移植肝排斥反应的同时,为预防肝癌肝移植术后复发、延长生存提供一种可能的方法.     

Cyclosporine and mycophenolate mofetil 48 hours before renal transplantation enables the use of low cyclosporine doses and achieves better graft function

Reducing calcineurin-inhibitor induced nephrotoxicity and simultaneously avoiding long-term side effects are desirable goals in renal transplantation. We examined the hypothesis that administration of cyclosporine and mycophenolate mofetil (MMF) 48 hours before renal transplantation allows reduction in the target cyclosporine C2 concentration, thus decreasing toxicity and improving graft function. We enrolled 80 kidney recipients in a single-center study comparing 2 cyclosporine-based protocols. Group I patients (n = 40) received a standard dose of cyclosporine (blood cyclosporine C2, 800-1500 ng/mL) with MMF and standard doses of corticosteroids. Group II patients (n = 40) were treated with a low dose of cyclosporine (blood cyclosporine C2, 450-800 ng/mL) and MMF plus low doses of corticosteroids after induction 48 hours before surgery with cyclosporine and MMF. Patient (97.5% vs 100%) and graft survivals (92.5% vs 95%) at 1 year were not different between the groups, although patients in group II experienced significantly fewer acute rejection episodes (10% vs 30%; P < .01). Delayed graft function occurred less often among group I than group II (17.5 vs 20%), but the difference was not significant. Graft function at 1 year was significantly better among group II (serum creatinine 1.31 vs 1.64 mg/dL and creatinine clearance 63 mL/min versus 47 mL/min; P < .05). We concluded that administration of cyclosporine and MMF 48 hours before renal transplantation allowed the safe effective use of low target C2 cyclosporine concentrations, enabling an early decrease in cyclosporine dose. These preliminary results indicated better 1-year graft function compared with the normal cyclosporine dose regimen.     

肝脏移植后的免疫损伤与热休克蛋白的表达

目的 探讨肝脏移植后的免疫损伤与几种主要分子伴侣(热休克蛋白)表达状况之间的内在规律。方法 34份移植后肝脏穿刺标本和10份正常肝脏标本，分为A(无排斥反应)组、B(轻／中度急性排斥反应)组、C(重度急性排斥反应)组、D(慢性排斥反应／肝纤维化)组、E(对照)组。进行HSP60、HSP70、HSP90、HO—1等四种分子伴侣免疫组织化学分析和图像分析。结果 B和C组各指标间无统计学差异，A、D、E与B、C组间有统计学差异。HSP60在移植肝脏中表达降低，排斥反应发生时高；HSP70和HSP90在移植肝脏中升高。HO—1肝脏移植后升高。结论 不同种类的热休克蛋白依据自身表达的特点对移植后的免疫损伤作出反应，体现了细胞的自我保护机制。     

Multiglycosidorum tripterygii, a new immunosuppressant, supresses coronary arteriosclerosis after heart transplantation.

BACKGROUND: Graft coronary arteriosclerosis is the major limiting factor for long-term survival after heart transplantation. In this study, we investigate the effect of multiglycosidorum tripterygii on graft coronary arteriosclerosis and platelet-derived growth factor A mRNA expression of transplanted hearts. METHODS: Three groups of Lewis rats (n = 7/Group) underwent heterotopic heart transplantation from Wistar-King donors and were treated with cyclosporine A (10 mg/ kg/day) for 60 days (Group A) or with multiglycosidorum tripterygii (30 mg/kg/day) for 60 days (Group B) or with cyclosporine A for the first 30 days and followed by multiglycosidorum tripterygii for another 30 days (Group C). Histological evaluations of rejection and coronary arteriosclerosis, as well as Northern blot analysis on graft platelet-derived growth factor A mRNA expression were made 60 days after transplantation. RESULTS: Morphometric results indicated no significant difference in rejection among three groups. However, the extent of graft coronary arteriosclerosis in Group B (1.12 +/- 0.21) and Group C (1.41 +/- 0.19) was significantly less than that seen in Group A (1.72 +/- 0.18) (p < 0.01 andp < 0.05, respectively). Furthermore, the incidence of diseased vessels was significantly less in Group B (29.5% +/- 7.8%) and Group C (42% +/- 9.1%) compared with Group A (69.1% +/- 11%) (p < 0.01 and p < 0.05, respectively). The expression of platelet-derived growth factor A mRNA of cardiac allograft was also significantly suppressed in Group B (25.4 +/- 6.2) and Group C (39.8 +/- 9.4), when compared with Group A (62.2 +/- 12.9) (p < 0.01 and p < 0.05, respectively). CONCLUSION: Multiglycosidorum tripterygii is superior to cyclosporine in prevention and attenuation of graft coronary arteriosclerosis and this efficacy is probably associated with the depressed expression of graft platelet-derived growth factor A mRNA in the multiglycosidorum tripterygii-treated groups.     

自体骨髓细胞诱导肝移植大鼠长期存活的机制探讨

目的 研究自体骨髓细胞诱导肝移植大鼠长期存活的可能机制.方法 雌性受体大鼠随机分成空白对照组(A组)、D-hanks液组(B组)、全骨髓细胞组(C组)、间充质干细胞组(D组).观察大鼠的中位生存时间(median survival time,MST)、肝功能、病理变化、Sry基因原位杂交和甲胎蛋白、白蛋白免疫组化双标检测观察自体骨髓细胞的分化情况.结果 C组、D组MST均>180 d(P<0.01);血肝功能指标C组、D组降低明显,有显著差异(P<0.01);C组、D组之间比较无显著差异(P>0.05);移植术后60 d C、D组均无明显的急性排斥反应;C组、D组Sry基因原位杂交和甲胎蛋白、白蛋白免疫组化双标检测呈阳性.结论 自体骨髓细胞能减少排斥反应、诱导大鼠肝移植术后长期存活,其中间充质干细胞在移植肝内诱导分化为肝细胞发挥肝细胞功能,是其中可能的机制.