慢性同种移植肾病大鼠早期MMP-9的表达

目的探讨基质金属蛋白酶-9(MMP-9)在大鼠慢性同种移植肾病(CAN)早期发病中的表达。方法按照国际标准的慢性同种移植肾病模型要求,以雄性Fisher(F344,RT1V1)大鼠作供体,雄性Lewis(LEW,RT1)为受体进行左肾原位移植。移植后第12周行功能学检查,按照Banff97工作分类标准进行组织学评价以及通过免疫组化检测MMP9和TGFβ1的表达。结果与对照组比较,实验组表现为移植肾肾小球不同程度的系膜增生和局灶节段性硬化,小管间质明显的单个核细胞浸润,局部小管萎缩,小动脉内膜轻度增厚和中层平滑肌细胞增殖并向内膜移行(P<0.01)。MMP-9在早期移植物内的表达增加,并与间质单个核细胞浸润相平行(r=0.77)。结论MMP-9作为一个重要因子参与CAN早期发病并在CAN单个核细胞浸润早期发病事件中起促进作用。     

肾移植致敏受者的配型研究及应用

目的:探讨人类白细胞抗原(HLA)配型在肾移植致敏受者中的应用及意义。方法:对32例PRA阳性的肾移植受者采用HLA交叉反应组(CREGs)配型原则选择供者,观察其供受者相配情况及移植效果。结果:按交叉反应组配型原则,供受者HLACREGS+DR0,1,2和3错配(MM)分别为8例(25％)、10例(31.25％)、12例(37.5％)和2例(6.25％),无4～6错配;而按传统的HLA-A,B,DR抗原配型原则,其0,1,2,3和4MM分别为2例(6.25％)、3例(9.38％)、10例(31.25％)、9例(28.12％)和8例(25％),其中0～1错配仅有5例。肾移植术后仅有9例发生急性排斥反应,经OKT_3治疗逆转,所有受者术后肾功能均恢复正常。结论:良好的HLA CREGs配型,可以显著提高供受者相配率,对减少致敏受者的排斥反应,提高移植肾存活率具有重要的意义。     

The Origin of Neointimal Smooth Muscle Cells in Transplant Arteriosclerosis from Recipient Bone-marrow Cells in Rat Aortic Allograft

In order to investigate the origin of neointimal smooth muscle cells in transplant arterio- sclerosis in rat aortic allograft, sex-mismatched bone marrow transplantation was performed from male Wistar rats to female Wistar rats. Four weeks after transplantation, the aortic transplant model was established by means of micro-surgery in rats. The recipients were divided into 4 groups: female Wistar-female Wistar aortic isografts, female SD-female Wistar aortic allografts, male SD-male Wis- tar aortic allografts, female SD-chimera Wistar aortic allografts. Eight weeks after transplantation, aortic grafts were removed at autopsy and processed for histological evaluation and immunohisto- chemistry. The results indicated that excessive accumulation of α-SMA-positive smooth muscle cells resulted in significant neointima formation and vascular lumen stricture in rat aortic allografts. Neointima assay revealed that the neointimal area and NIA/MA ratio of transplanted artery were sig- nificantly increased in all of aortic allograft groups as compared with those in aortic isograft group (P<0.01). Neointimal smooth muscle cells were harvested from cryostat sections of aortic allograft by microdissection method. The Sry gene-specific PCR was performed, and the result showed that a dis- tinct DNA band of 225 bp emerged in the male-male aortic allograft group and chimera aortic al- lograft group respectively, but not in the female-female aortic allograft group. It was suggested that recipient bone-marrow cells, as the origin of neointimal smooth muscle cells, contributed to the pathological neointimal hyperplasia of aortic allograft and transplant arteriosclerosis.     

Factors affecting the long-term renal allograft survival

Background  In the past decades, the one-year graft survival of cadaveric renal allografts has been markedly improved, but their long-term survival has not kept pace. The attrition rate of renal allografts surviving after one year remains almost unchanged. The causes for late graft loss are multiple. The aim of this study was to analyze the predictive factors that impact long-term survival of grafts after kidney transplantation.

Methods  We retrospectively analyzed 524 kidney transplantation patients who were treated in our hospital between January 1991 and January 2000, including 254 patients who had lived more than 10 years with normal graft function (long survival group), and 270 cases whose renal graft had survived less than 10 years (control group). Specifically, we analyzed 10 factors that may potentially affect graft survival by both univariate and Logistic model multivariate analyses to pinpoint the independent risk factors. 

Results  Univariate analyses showed that no significant differences existed in the age or gender of recipients, dialysis time, lymphotoxin levels, or cold ischemia time between the two groups. However, the ratio of delayed graft function and acute rejection, and the uric acid levels of patients in the long survival group were significantly lower than those in the control group (P <0.01). Furthermore, we found that the concentration of cyclosporin A at one year after transplantation and the histocompatibility antigen match of donor-recipients for patients within the long survival group were significantly higher than those in the control group (P <0.01). Furthermore, multivariate analyses showed that these four factors were independent risk factors that impact patient survival.

Conclusions  The ratios of delayed graft function and acute rejection, the concentration of cyclosporin A at one year after transplantation, and serum uric acid levels are very important factors that affect the long-term survival of renal grafts.

     

心脏死亡供肾肾移植19例临床分析

目的探讨终末期肾病患者接受DCD供者肾肾移植后的恢复情况及DCD供体对受者及移植物术后的影响．方法分析昆明市第一人民医院19例终末期肾病患者接受DCD捐献后肾移植的临床资料,对其术前、术后的诊疗及血肌酐（sCr）、移植物及受者存活情况进行回顾性分析．结果19例患者手术全获成功．术后无1例原发性移植物无功能（PNF）病例发生．供体sCr正常的6例,供给11例受体中7例发生肾功能延迟恢复（DGF）,发生率为63．6％;供体sCr（184～504μmol/L）异常的8例,供给的8例受者均发生DGF,发生率为100％;19例受体中有1例（5．3％）发生超急性排斥反应,移植肾丢失;有2例术后分别出现严重尿瘘和严重骨髓移植．18例受者接受随访,随访时间为3～12个月,平均7个月,移植肾功能完全恢复正常．结论DCD是目前我国器官来源的重要部分,是解决我国器官移植面临的器官短缺的一个非常有潜力的办法,并且有着较好的短中期预后．     

应用国产雷帕霉素延长肾脏移植物存活的实验研究

目的探讨国产雷帕霉素(rapamycin, RPM)及其与环孢素A(CsA)联合应用延长大鼠移植肾存活的效能.方法肾移植大鼠分为对照组、RPM组、CsA组、RPM 小剂量CsA组和RPM 大剂量CsA 4个治疗组.观察移植肾存活时间、血肌酐浓度和移植肾病理改变.结果与对照组相比,RPM组、CsA组、RPM CsA组移植肾存活时间均显著延长(P<0.01),其中,RPM 小剂量CsA组移植肾存活时间最长(69.2±10.3)d,术后15、30d组血肌酐浓度最低,未见明显的肾小管空泡变性.结论国产雷帕霉素可有效延长大鼠移植肾存活时间;与小剂量CsA合用可避免严重肾中毒,进一步延长移植肾的存活.     

紫杉醇防治移植动脉硬化作用的实验研究

目的采用大鼠胸腹主动脉移植模型，探讨紫杉醇防治移植动脉硬化的作用。方法Wistar大鼠40只，SD大鼠16只，按供、受鼠不同分为4组：A组为假手术组；B组为同品系移植（Wistar to Wistar）组；C组为单纯异品系移植（Wistar to SD）组；D组为异品系移植（Wistar to SD）紫杉醇干预组。移植术后30天切取移植动脉，采用光镜和电镜定性观察，并用计算机图像分析系统定量观测紫杉醇对移植动脉的影响。结果与A组比较，术后30天B组动脉壁无增厚、血管内膜未见明显增生；C组内膜增生明显，内外膜有大量炎症细胞浸润。紫杉醇则显著抑制移植动脉内膜增生，并减轻移植动脉内外膜炎症细胞浸润，防止管腔狭窄。结论紫杉醇可明显抑制异品系大鼠移植动脉内膜增生和炎症细胞浸润，防治移植动脉硬化。     

Clinical significance of hemorrheological parameters in patients with renal transplantation

Hemorrheological studies carried out on 14 recipients of cadaveric kidney transplant regularly before and after transplantation showed very significant differences in plasma viscosity in the period of stable function after successfully transplanted kidney, dialysis before transplantation, and renal rejection (P<0.00l). Significant difference was observed in fibrinogen level in cases with stable normal function and rejection (P<0.05,). Significant positive correlation was found between plasma viscosity and fibrinogen level in patients on dialysis as well as after transplantation. The results in this series showed that abnormal changes in hemorrheology respond to the development of rejection episode and deterioration of renal function in patients after receiving cadaveric renal allografts.     

活体肾移植体液性排斥反应与抗HLA抗体的研究

目的研究活体肾移植术后体液性排斥反应与抗HLA抗体及其特异性的关系。方法87例活体肾移植患者,分别于肾移植术前1天及术后6个月行流式细胞法群体反应抗体检测(Flow PRA screening test)。同时应用独立抗原免疫磁珠分析法(LAB Single antigen analysis)检测抗HLA抗体的特异性。全部患者于术中、术后两周、术后6个月和1年4个时间段进行移植肾穿刺病理检查。结合患者一般情况、病理诊断及抗HLA抗体检测结果,在体液性排斥反应发生率、预后、相关抗体种类及特异性等方面进行回顾性分析。结果87例患者中,群体反应抗体(PRA)术前1天检测结果均为阴性。术后6个月时28例(32.2%,28/87)为阳性。其中,15例(53.6%,15/28)为非供体特异性抗HLA抗体;13例(46.4%,13/28)存在供体特异性抗体。病理结果提示,供体特异性抗体患者中11例(84.6%,11/13)在术后6个月内出现了严重的抗体介导的体液性排斥反应;术后1年时仍然有5例持续存在体液性排斥,移植肾3年内完全丧失功能,恢复到规律透析状态。非供体特异性抗体患者术后无体液性排斥反应发生。术后抗HLA抗体阴性组与阳性组3年移植肾存活率分别为96.6%和75.0%。结论活体肾移植患者抗HLA抗体的出现与术后急性体液性排斥反应的发生明显相关,特别是术后出现供体特异性抗HLA抗体的患者急性体液性排斥反应的发生率更高,预后更差。肾移植术后严密监测抗HLA抗体的出现对于及时调整免疫抑制方案改善移植肾长期存活具有重要意义。     

犬肾移植致敏模型的建立

目的建立犬致敏后的肾移植动物模型。方法雄性家犬各6条配对作为淋巴细胞供受体,采用小剂量（0.4×10^7-1.2×10^7个/kg）淋巴细胞多部位多次输注的方法诱导致敏。用补体依赖的淋巴细胞毒（complement deendent cytotoxicity,CDC）和混合淋巴细胞培养（mixed lymphocyte culture,MLC）试验进行检测。当CDC转为阳性和MLC显示反应淋巴细胞增殖明显活跃后,淋巴细胞供受体犬间进行交叉肾移植,术后定期ECT动态显像观察移植肾功能变化;分批摘取移植肾行病理检查,观察排斥反应发生情况。结果实验犬均在输注淋巴细胞3-4次后CDC转为阳性,MLC显示反应淋巴细胞增殖明显活跃。肾移植术后4天ECT检查有4条致敏犬移植肾血流灌注下降,肾功能出现损害,摘取的3只移植肾病理检查表现为抗体介导的排斥（延缓）或急性排斥反应;对照组未见异常。术后7天,致敏犬移植肾均因急性排斥失功;对照组3条犬有2条出现肾功能损害,病理检查示急性排斥反应。结论小剂量淋巴细胞多部位多次输注可以诱导出家犬的免疫致敏状态。致敏犬肾移植术后移植肾排斥反应出现较早,肾功能损害更严重。     

Establishment of a sensitized canine model for kidney transplantation

Objective:To establish a sensitized canine model for kidney transplantation. Methods: 12 male dogs were averagely grouped as donors and recipients. A small number of donor canine lymphocytes was infused into different anatomic locations of a paired canine recipient for each time and which was repeated weekly. Specific immune sensitization was monitored by means of Complement Dependent Cytotoxicity (CDC) and Mixed Lymphocyte Culture (MLC) test. When CDC test conversed to be positive and MLC test showed a significant proliferation of reactive lymphocytes of canine recipients, the right kidneys of the paired dogs were excised and transplanted to each other concurrently. Injury of renal allograft function was scheduled determined by ECT dynamic kidney photography and pathologic investigation. Results:CDC test usually conversed to be positive and reactive lymphocytes of canine recipients were also observed to be proliferated significantly in MLC test after 3 to 4 times of canine donor lymphocyte infusions. Renal allograft function deterioration occurred 4 d post-operatively in 4 of 6 canine recipients, in contrast to none in control dogs. Pathologic changes suggested antibody-mediated rejection (delayed) or acute rejection in 3 excised renal allograft of sensitized dogs. Seven days after operation, all sensitized dogs had lost graft function, pathologic changes of which showed that the renal allografts were seriously rejected. 2 of 3 dogs in control group were also acutely rejected. Conclusion：A convenient method by means of repeated stimulation of canine lymphocyte may induce specific immune sensitization in canine recipients. Renal allografts in sensitized dogs will be earlier rejected and result in a more deteriorated graft function.     

Transplantation of kidneys from living related donors. Comparison with cadaveric kidney transplantation

In a series of 271 transplantations of renal allografts, performed over 10 years, the rates of graft survival, patient survival, and morbidity in the recipients of allografts from living related donors (47 allografts) have been compared with those in the recipients of cadaveric allografts (224 allografts). The one-year graft survival rates were 88% for allografts from living related donors (100%, if these were HLA-identical) and 55% for cadaveric allografts, while the patient survival rates were 97% and 87%, respectively, in the same period. Morbidity rates (expressed as the number of days spent in hospital) for recipients of allografts from living related donors were approximately 50% of those for recipients of cadaveric grafts. Complications in the living related donors were minimal, and acceptable. It is concluded that transplantation of allografts from living related donors has many advantages over transplantation of cadaveric kidneys, and is a valuable adjunct to a cadaveric renal transplantation programme. Greater use of living related kidney donors should be encouraged in Australia.     

肾移植术后BK病毒感染研究进展

BK病毒是一种隶属于多瘤病毒种属的DNA病毒,在正常人群中的感染率高达90%。然而,仅在免疫功能受损的个体中引发相关疾病,以接受移植术后服用免疫抑制药物的患者最为显著。BK病毒的持续复制被认为是导致肾移植受者移植肾功能受损的重要因素,而由其引发的BK病毒性肾病(BKVN)则已成为肾移植失败的重要原因之一。应用尿细胞学染色和检测尿液或血液中BK病毒DNA载量等方法可对BK病毒感染和BKVN进行监测并早期诊断,增加了早期治疗的成功率。然而,由于缺乏特异性的抗病毒药物,BKVN的治疗仍然非常困难。该文针对肾移植术后BK病毒感染的流行病学、发病机制、实验室检测方法以及治疗策略的研究予以综述。     

移植肾转化生长因子β1与远期肾功能的关系

目的：探讨移植肾内转化生长因子β1（TGF-β1）的表达与远期肾功能的关系。方法：对168例肾功能正常的肾移植患者检测尿TGF-β1含量，所得相对浓度为151.3~521.7（pg/mg.Cr），分别选出浓度最高和最低的各20例患者构成A组、B组。检测2组血TGF-β1浓度；2组分别有15例和12例患者进行了移植肾穿剌活检，通过免疫荧光检测活检组织中TGF-β1蛋白的表达；3年后，比较2组肾功能有无差异，对肾功能不全者作移植肾穿剌活检，明确移植肾在病理上是否为非特异性损害（即慢性移植肾肾病的改变）。结果：2组患者血TGF-β1浓度没有明显差异；A组活检显示在肾小管基底膜等处有大量的TGF-β1表达，其免疫荧光强度为（10.52±2.36）×106，明显大于B组［（6.48 ±1.53）×106］；3年后，A组肌酐清除率（Ccr）减少了（17.6±6.9） ml/min、有7例患者肾功能不全，与B组比较有显著差异（B组分别为（6.3±4.4） ml/min和1例）；肾功能不全的患者，移植肾穿剌活检均证实为慢性移植肾肾病。结论： TGF-β1与慢性移植肾肾病的发生有着密切的关联；肾移植后检测尿TGF-β1对远期肾功能具有预测作用；移植肾TGF-β1高表达者远期肾功能差。     

Variation of haemorheological parameters in renal and pancreatic rejection

Summary Abnormalities of haemorheology were found in animal and human recipients of kidney and/or pancreas allografts during rejection episodes. Thirteen diabetic canines received solitary pancreatic transplantation and another 13 diabetic and uremic canines underwent combined pancreas and kidney transplantation. Determination of haemorheological parameters was performed before and after operation respectively. During rejection episodes of kidney or pancreas allografts, the values of plasma viscosity, blood reductive viscosity and fibrinogen were significantly higher than those without rejection. On the basis of animal experiments, the determination of haemorheological parameters had been performed on 33 patients (30 receiving renal tramsplant, 2 pancreatic transplant alone and the remaining 1 conbined renal and pancreatic transplant). Consecutive mornitoring on these patients showed that a rise in the values of plasma viscosity, blood reductive viscosity and fibrinogen could be demonstrated during rejection episodes. The changes appeared one to three days prior to clinical manifestations and were in accordance with the termination of rejection. Our studies suggest that variation of haemorheological parameters are associated with rejection and the abnormal haemorheology may be an essential factor contributing to graft dysfunction. Moreover, the use of these assays will be beneficial to early diagnosis and better management of rejection in the future.     

CMV-PP65抗原检测在预防肾移植术后巨细胞病毒感染的临床价值

目的 探讨巨细胞病毒PP65抗原（CMV-PP65）检测在预防肾移植术后巨细胞病毒（CMV）感染的临床价值.方法 采用间接免疫荧光法和酶联免疫捕获法检测52例肾移植受者术后CMV-PP65抗原和CMV-IgM抗体.结果 52例肾移植受者中CMV-PP65抗原检测阳性25例,CMV-IgM抗体检测阳性17例,随访3~6个月,共15例发生CMV病,CMV-PP65抗原检测均阳性,CMV-PP65抗原检测的敏感度及阴性预测值均高于CMV-IgM抗体检测;25例患者CMV-PP65抗原检测术后首次出现阳性的平均时间为4.2±3.3周,17例患者CMV-IgM抗体检测术后首次出现阳性的平均时间为8.7±5.8周,两者比较有显著差异性（P〈0.05）.结论 CMV-PP65抗原检测可早期排除和预防肾移植术后CMV感染,值得临床推广应用.     

TGF-β1与慢性移植肾肾病关系的临床研究

目的:探讨移植肾转化生长因子β1(TGF-β1)与慢性移植物肾病的关系。方法:对152例肾功能正常的肾移植患者检测尿TGFβ1含量,所得相对浓度为157.0～520.7(pg/mg·Cr),分别选出浓度最高和最低的各25例患者构成A组、B组。检测两组血TGF-β1浓度;两组中分别有15例和12例患者进行了移植肾穿剌活检,比较活检组织中TGFβ1mRNA和TGFβ1蛋白的表达;3年后,比较两组肾功能有无差异,对肾功能不全者作移植肾穿剌活检,明确移植肾在病理上是否为非特异性损害(即慢性移植肾肾病的改变)。结果:两组患者血TGFβ1浓度没有明显差异;A组活检组织中TGF-β1mRNA表达量和TGFβ1免疫荧光强度分别为(1.38±0.28)×106和(10.80±2.31)×106,明显高于B组,后者分别为(0.88±0.25)×106和(6.41±1.57)×106;3年后,A组肌酐清除率(Ccr)为(79.2±11.2)ml/min、有7例患者肾功不全,与B组比较有显著差异;肾功不全的患者,移植肾穿剌活检均证实为慢性移植肾肾病。结论:TGF-β1与慢性移植物肾病的发生有着密切的关联;肾移植后检测尿TGF-β1对远期肾功能具有预测作用。     

供肾质量和受者体质量与移植肾远期功能的关系

目的探讨肾移植术后供肾质量、受者体质量及二者比值与远期肾功能的关系。方法对108例肾移植手术病人的供肾质量、受者体质量及二者的比值与受者术后3年血肌酐值进行线性相关分析。结果供肾质量与术后3年血肌酐值呈负相关(P<0.05);受者体质量与术后3年血肌酐值呈正相关影响(P<0.01);供肾质量及受者体质量的比值与术后3年血肌酐值呈负相关(P<0.01)。结论供肾质量、受者体质量及二者的比值与远期移植肾功能的关系应引起重视,体质量较大的受者应尽量选择较重的供肾相配。     

Treatment of chronic dysfunction of transplantation kidney in rats —by tanshinone, lysimachiae combined with mycophenolate mofetil or cyclosporine alone

Objective: To evaluate the relationship between chronic kidney dysfunction after transplantation and chronic vascular rejection (CVR), and to evaluate the efficacy and safety of Tanshinone (Tan) and Herba Lysimachiae (Lys) combined with Mycophenolate Mofetil (MMF) to fight against CVR, and to reduce the incidence of chronic dysfunction in rat renal transplantation model.Methods: Sixty-five male SD rats as donors and sixty-five male Wistar rats as recipients were used. The recipients were divided into five Groups, including Group A: Lys + Cyclosporine A (CsA), Group B: Tan+CsA, Group C: MMF+CsA, Group D: CsA, and Group E: normal saline. Kidney function and morphological changes were assessed at 2, 4, and 6 weeks after transplantation. All sections of renal grafts were stained with monoclonal antibodies (McAB),. including major histocompability complex class II (OX-6), lymphocyte function antigen-1 (CDllb/CD18), intercellular adhesive molecular-1 (IA₂₉), CD₈ ⁺ (OX-8), and proliferation cell nuclear antigen (PCNA) (5A10, IgGlk) were used. Results:The two control groups developed typical chronic rejection episodes, and the histologic feature indication of kidney chronic rejection includes loss of renal units and presence of an obliteration arteriopathy involving large renal arteries. These were associated with serum levels of BUN and SCr increased and different degrees of glomerulosclerosis. Their mean survival time was lower than that of other groups. By contrast, serum levels of cytokine in control groups was significantly increased when compared with group B and C (P<0.05). Both group B and C had minimum changes in glomeruli and arteries, and expression levels of PCNA on the glomeruli and tubular cells were higher than those of other groups (P<0. 05). However, there was no significant difference (P>0.05) between group B and C.Conclusions: CVR may activate the risk of the factor responsible for the development of graft chronic dysfunction that causes slow, progressive destruction of the transplanted kidney. Tanshinone was extracted from Salvia miltiorrhiza and purified for use in medicine. Especially when Tanshinone combined with a low-dose of CsA, it may fight against the CVR by inhibiting cell infiltration, and improving microcirculation of the graft, and thus the incidence of CVR is reduced. It is suggested that Tanshinone can be applied to treat patients with chronic renal dysfunction when used in combination with a low-dose Cyclosporine. This work was supported by the Sciences Foundation of the Ministry of Health Grants (No. C 0103)     

供肾质量和受者体质量与移植肾远期功能的关系

OBJECTIVE: To investigate the relationship of the renal allograft weight and the recipient's body weight with allograft function after transplantation. METHODS: The correlation of the renal allograft weight, the recipient's body weight, the ratio of the allograft weight to the recipient body weight and the mean serum creatinine (sCr) 3 years after transplantation were measured in 108 kidney recipients. RESULTS: The allograft weight was inversely correlated with the mean sCr 3 years after transplantation (P<0.01). CONCLUSION: The renal allograft weight, recipient's body weight and the ratio of allograft weight to recipient's body weight are important indicators of the long-term allograft function after transplantation, and recipients with greater body weight should receive allografts of higher weight.