大鼠喉移植的应用解剖学研究

目的 :观察和研究大鼠颈部肌肉、喉的动脉、静脉及神经支配 ,探讨喉移植血管、神经的选择及吻合模式。方法 :对 5 0只Wister大白鼠进行解剖观察 ,分别测量各主要血管及神经的直径和长度。结果 :大鼠喉的主要动脉为甲状腺上动脉 ,直径 ( 0 68± 0 0 4)mm ,长度 ( 4 74±0 94)mm ,颈总动脉直径 ( 1 5 3± 0 16)mm ,静脉主要是颈外静脉 ,直径 ( 1 82±0 2 3 )mm ,喉返神经为支配喉声带运动的神经 ,直径 ( 0 5 2± 0 0 6)mm。结论 :大鼠喉移植宜选择双侧带有甲状腺上动脉的颈总动脉与受体鼠的颈总动脉、颈外静脉重建血液循环 ,供体鼠的喉返神经可与受体的副神经、舌下神经进行吻合 ,恢复神经支配功能     

移植免疫耐受研究获新进展

中山医科大学肾脏病临床研究所博士研究生史伟等在导师张仕光教授的指导下，根据自身免疫耐受的形成机制，采用逆转录病毒载体介导的基因转移技术，将供鼠主要组织相容性抗原复合体类抗原基因，在体外转移到不同品系的受体鼠T淋巴细胞内表达，然后将此基因修饰的受体T淋巴细胞回输受体鼠胸腺内，以皮肤移植作为器官移植的观察模型，成功地诱导了受体对洪体移植物的特异性免疫耐受，对防止移植物发生免疫排斥反应具有重要意义。此研究为今后的器官移植研究建立了一个符合生理状态的诱导免疫耐受的观察模型。这对于诱导同种异体器官移植免疫…     

透射电镜用于供体喉保存的评价

目的:研究大鼠供体喉的保存,探讨鼠喉保存的评价方法及保存时间。方法:采用Wister大白鼠,切取供体喉后用UW氏液进行灌洗,并低温保存至不同时段,分别通过病理光学及透射电镜观察其形态学改变。结果:病理光镜检查在移植喉保存60 h后,才能发现大鼠喉组织的缺血性改变,透射电镜可以在鼠喉保存24 h后发现细胞的超微结构改变,28 h后供体喉大部分组织出现细胞核固缩和凋亡。结论:透射电镜能及早地反映喉体组织缺血和坏死情况,优于病理光镜检查,UW氏液保存移植喉可达26 h。     

建立裸鼠FaDu喉咽癌原位种植瘤模型及其CT和病理研究

目的建立Fa Du喉咽癌原位种植瘤模型并对其CT及病理诊断进行分析。方法 Fa Du肿瘤细胞株复苏培养传代建立荷瘤裸鼠,在裸鼠喉咽部注入,建立Fa Du喉咽癌原位种植瘤模型。结果裸鼠喉咽部接种Fa Du肿瘤细胞悬液,2周后可成功建立裸鼠喉咽肿瘤模型,成瘤率100%(30/30),动物CT检查能明确发现肿瘤占位,HE染色证实肿瘤组织为中至高分化鳞状细胞癌。结论 Fa Du细胞悬液喉咽癌接种建立的裸鼠头颈部肿瘤模型具有建模周期短,稳定性好、易于重复、移植瘤成功率高、操作简单等特点。     

二甲基肼诱导昆明小鼠大肠癌的研究

目的: 选择性去除骨髓移植物中异基因反应性淋巴细胞,特异性抑制移植物抗宿主病(GVHD).方法: 用携带有FasL基因的重组腺病毒转染Balb/c小鼠来源的树突状细胞(dendritic cells, DC),并与C57BL/6小鼠骨髓细胞移植物共培养,把经过这种处理的骨髓细胞移植物移植给Balb/c受体小鼠(C57BL/6→Balb/c小鼠GVHD 模型 ,H-2b→H-2d),然后观察、比较各组GVHD表现.结果: 致死剂量照射的受体鼠在接受经FasL-DC处理的供体骨髓细胞移植后,没有出现明显的GVHD表现,生存期显著延长,3个月时生存率80%以上.但对照组2周后均出现了明显的GVHD症状,腹泻、脱毛和靶组织淋巴细胞浸润等,生存期没有超过30 d.结论: 转染FasL基因的DC可有效去除骨髓移植物中异基因反应性T淋巴细胞,移植用这种方法处理过的骨髓,能够有效抑制GVHD的发生.     

游离空肠移植重建下咽及颈段食管的临床应用及初步评价

自1986年9月至1988年9月,我院用游离空肠移植方法重建下咽及颈段食管癌术后缺损,共完成7例.男5例,女2例;年龄自38岁～67岁,平均57.3岁.Ⅲ期6例(T_3N_05,T_3N_11),Ⅳ期1例(T_3N_2).梨状窝癌累及喉5例,环后癌1例,颈段食管癌1例.全喉咽及颈段食管切除6例,颈段食管切除(保留喉)1例.1例发生肠段坏死,血管吻合成功率为85.7%(6/7).3、5年生存率分别为57.1%(4/7)及33.3%(1/3).介绍了手术要点,该术式的优点及局限性.     

bcr-abl反转录病毒介导的慢性粒细胞白血病样小鼠二次移植模型的建立

目的:构建bcr-abl反转录病毒介导的小鼠慢性粒细胞白血病(chronic myeloid leukemia,CML)样二次移植模型,为深入研究CML的发病和治疗机制奠定可靠的基础.方法:收集已成功建模的由bcr-abl反转录病毒介导的CML样模型小鼠的骨髓细胞或脾细胞,经尾静脉输注入经亚致死剂量γ射线(450 cGy)照射的同种雌性受体鼠,进行造血组织的形态学观察、Y染色体及其特异性性别决定基因Sry的检测以及bcr-abl mRNA和蛋白水平表达的检测,分析鉴定CML样小鼠二次移植模型情况.结果:移植后12 d,肉眼可见明显的脾结节形成,连续切片可见成堆脾集落.移植4～5周后,在雌性受体小鼠骨髓中检出相似Y染色体及Y染色体特异性Sry基因,骨髓和脾脏中检出bcr-abl融合基因.移植10～12周后外周血白细胞水平升高至正常未处理小鼠的4～8倍,粒系细胞水平明显升高至(20～32)×109个/L,外周血涂片幼稚细胞占10%～20%;骨髓涂片结果显示粒系细胞显著增多,易见幼稚细胞,肝脾也可见白血病细胞浸润;同时,在肝脏中检测出bcr-abl融合基因,骨髓中检测出bcr-abl融合蛋白的表达.移植18周后,外周血涂片幼稚细胞占30%以上,小鼠发生急性变,最终因肺出血相继死亡.结论:本研究成功建立了bcr-abl反转录病毒介导的小鼠CML样二次移植模型,该模型可用于后续CML信号通路和治疗机制的研究.     

大鼠喉移植供体喉的体外保存研究

背景与目的：喉作为一个复合器官,供体喉的体外保存时间不能超过3小时。本研究探讨供体喉的最佳保存方法及时间。方法：在大鼠喉移植实验中,采用HCA液（hyperlonie citrate adeuinc solution）和UW氏液（university of winstousin solution）分别进行供体喉原位灌注,观察低温保存后喉体的连续形态学改变,比较两种保存液的保存效果。结果：HCA液保存20小时后,移植喉出现粘膜上皮、血管内皮、喉内肌等组织细胞的超微结构变化,线粒体肿胀、崩解,细胞核染色质边集,22小时后细胞发生核固缩,继而出现凋亡现象;UW氏液保存24小时后才开始出现喉体各组织细胞缺血、坏死改变。结论：UW氏液对供体喉的保存效果优于HCA液,能保存喉体达24小时。     

肝细胞移植治疗大鼠急性肝衰竭的疗效观察

目的观察肝细胞移植治疗大鼠急性肝衰竭的疗效, 探讨原代肝细胞提取方法, 观察体外培养的大鼠肝细胞生存情况。方法 利用胶原酶消化法分离大鼠肝细胞, 用D-氨基半乳糖(D-GalactosamineN, D-GalN)制作大鼠急性肝功能衰竭模型, 并以此模型为基础行大鼠同种异体肝细胞移植实验。结果 分离的肝脏实质细胞(Hepatocyte)的存活率在87%-95%。平均每只大鼠肝脏可分离出7.8×10⁷个肝脏实质细胞。肝细胞移植组大鼠7天的存活率为54.2%(13/24), 对照组8.3%(2/24), 移植组大鼠存活率与非移植组大鼠存活率之间具有显著性差异(P＜0.005)。结论 经腹腔移植同种异体肝细胞可明显改善D-GalN诱导的急性肝衰竭大鼠的存活率, 腹腔肝细胞移植是治疗急性肝衰竭安全有效的方法。     

声门上喉鳞癌nm23H1、表皮生长因子受体表达及其与颈淋巴结转移和预后的关系

目的:研究声门上喉癌中nm23H1,表皮生长因子受体(epidemal growth factor receptor,EGFR)表达及其与颈淋巴结转移和预后的关系.方法:回顾分析77例声门上喉鳞癌和23例癌旁正常喉粘膜标本.应用免疫组化技术研究nm23H1,EGFR的表达.结果:声门上喉鳞癌中nm23H1,EGFR的阳性表达率为76.62%和72.73%.声门上喉鳞癌和癌旁正常喉粘膜中nm23H1和EGFR表达具有显著差异(P＜0.05).nm23H1表达在颈淋巴结转移阳性组中显著降低(P＜0.05),且与临床分期间有显著差异(P＜0.05).EGFR表达在声门上喉癌与癌旁正常喉粘膜,颈淋巴结转移阳性与阴性组间无显著差异.Kaplan-Meier与Cox模型分析显示nm23H1,EGFR表达均不能影响生存和复发.结论:nm23H1表达在颈淋巴结转移中显著降低,提示有可能将之作为声门上喉癌颈淋巴结转移的预测指标.     

同种异体松质骨移植动物模型的建立和评价

目的建立同种异体松质骨移植动物模型，并对其术后免疫反应进行评价。方法取近交系Lewis大鼠和Fisher344大鼠各15只，切取双侧股骨下端干骺端（松质骨）作为移植骨块。另取Fisher344大鼠30只作为受体，随机分为A组（同基因移植组）、B组（同种异基因移植组），每组15只。移植手术为在受者双侧股后肌袋内植入移植骨块，术后2、4、8周分批取材检测，观察术后受者的淋巴细胞刺激增殖效应、T淋巴细胞亚群CD4／CD8比值变化以及移植块的组织学改变。结果B组的淋巴细胞刺激增殖指数及淋巴细胞亚群CD4／CD8的比值在术后的各个时期均比A组要高（P〈0．05），在术后第4周达到高峰；组织学检查表明B组移植骨块的炎性细胞浸润较多，成骨较少。结论本研究建立的同种异体松质骨移植实验动物模型术后发生了免疫反应并可以检测，可以作为下一步研究降低同种异体松质骨免疫原性方法的实验动物模型。     

Nephron-sparing surgery of a low grade renal cell carcinoma in a renal allograft 12 years after transplantation

Renal cell carcinoma (RCC) occurring in renal allografts after cadaveric kidney transplantation has rarely been observed. RCC accounts for 2.3% of all malignancies in the general population, but up to 4.8% of malignancies in renal transplant recipients. Most have been reported in the patient's own diseased kidneys, whereas RCC in the renal allograft occur in only 10%. Here, we describe an organ-preserving surgical technique of a malignant renal tumor in a kidney allograft using a harmonic scalpel (Ultracision) for tumor enucleation. Furthermore we demonstrate by DNA microsatellite analysis the tumor's genetic origin as donor related. Collectively, we suggest that patients with a well defined low grade RCC in the kidney allograft and altogether low malignancy and good allograft function should only undergo an organ-preserving procedure and short-term postoperative screening.     

肾移植患者术后肿瘤发病特点及预后

背景与目的由于肾移植患者长期应用免疫抑制剂,与普通人群相比更易发生肿瘤。本研究中我们主要分析肾移植患者术后发生肿瘤的临床特点和预后,并评价根治性手术(radicalsurgery,RS)对其预后的影响。方法对本院1987年11月～2004年5月行肾移植手术的2160例患者进行回顾性分析,分析肾移植术后肿瘤的发生时间、肿瘤类型及生存时间等,总结肾移植术后肿瘤的发病特点。按是否行RS将肿瘤患者分为两组,对比研究RS对其预后的影响。结果2160例肾移植手术的患者中33例术后发生肿瘤,以消化系统肿瘤为主(33.3%)。10例行RS治疗(RS组)的患者中位生存时间为41.5个月;23例未行RS治疗(非RS组)者中位生存时间为6.0个月。两组20个月生存率分别为70.0%和13.0%。结论肾移植患者比普通人群更易发生肿瘤,肿瘤类型与普通人群所患不同,以肝癌、皮肤癌、淋巴瘤、甲状腺癌等为主。早期发现、早期治疗,尤其是病情允许行根治性手术者,近期疗效较好,远期效果有待进一步观察。     

Incidence of hyperthyroidism after unrelated donor allogeneic stem cell transplantation

We report on three patients who developed overt thyrotoxicosis after volunteer unrelated donor bone marrow transplantation for Philadelphia chromosome positive chronic myeloid leukemia shortly after the onset of chronic graft versus host disease. In all three cases, the etiology of hyperthyroidism is likely to be a combination of toxic factors and an immune process. Systematic evaluation of thyroid function tests in 97 unrelated allograft recipients from our center who survived at least 100 days from stem cell or bone marrow transplantation for hematological diseases gave a rate of overt thyrotoxicosis at 3.1% in this cohort.     

Biological testing of antilymphocyte sera and globulin in the baboon

A variety of antilymphocyte sera (ALS) and globulins (ALG) as well as other adjuvant compounds (procytoxide, PCO) were evaluated in 40 adult chacma baboons housed at the Roswell Park Surgical Primate Center. Animals were characterized in terms of human erythrocyte and leukocyte compatibility and then entered into the test evaluation program. Paired animals exchanged first- and second-set skin grafts and subsequently renal allografts with and without therapy. Appropriate interval periods elapsed between each allograft test period. Baboon–goat ALS and human thymus ALG were all significantly effective in prolongation of first- and second-set skin grafts as well as renal allografts. PCO slightly prolonged survival of sensitized renal allograft recipients. Serological (immunochemical) studies, level of peripheral lymphocytes, and prior in vitro tests all failed to correlate with the observed effectiveness of the ALS and ALG preparations noted in the present series of baboons. The baboon thus appears to be a vital as well as valid in vivo model for testing and evaluation of ALS and ALG preparations.     

Papillary microcarcinoma of the thyroid gland in renal transplant patients

Among organ transplant recipients there is a world wide increase in the number of de novo tumors as well as a decrease in the time of the first appearance after the transplantation. Between 1973 and the 31st of August 1999 1709 cadaver renal allograft transplantations were performed in our Department. Four thyroid cancers were detected among the renal transplanted patients. Two of them proved to be papillary microcarcinomas. Although the elevated risk of thyroid cancers is well established in the literature papillary microcarcinomas have never been reported before in an immunosuppressed patient. Authors highlight that the thyroid gland should always be carefully checked in organ transplant recipients, since better survival might be achieved even in the immunosuppressed population. Metastatic tumor is relatively benign which is in correlation with the literature, but there has been little experience in organ transplanted patients so far.     

The current status of reduced-intensity allogeneic hematopoietic stem cell transplantation for multiple myeloma.

Of all the treatment modalities employed to control multiple myeloma, only allogeneic hematopoietic stem cell transplantation is potentially curative, due in large part to a graft-versus-myeloma (GVM) effect. Whereas patients who receive either allogeneic or autologous stem cell transplants for multiple myeloma have similar 3-5-year survival, only allograft recipients appear to enjoy long-term disease-free survival. High transplant-related mortality (TRM) associated with allogeneic stem cell transplantation is currently the major limitation to wider use of this potentially curative modality. This high mortality has been the major impetus for exploration of reduced intensity conditioning (RIC) regimens designed to allow engraftment of allogeneic stem cells. With follow-up now extending to 7 years, it is clear that when compared to myeloablative transplants, RIC allografts are associated with lower TRM; however, reduced mortality comes at a cost of higher rates of disease progression and relapse. Strategies designed to improve the therapeutic index of allografts include the use of more intensive, yet still non-myeloablative conditioning regimens, tandem autologous plus RIC allografts, peripheral blood cells rather than bone marrow, graft engineering to improve the GVM activity while reducing graft-versus-host disease, post-transplant maintenance and targeted conditioning therapies such as bone-seeking radioisotopes.     

Islet cell function in long-term surviving primates after segmental pancreatic allotransplantation

Islet cell function was studied in pancreatectomized primates with functioning segmental pancreatic allografts more than 100 days after transplantation. Segmental allograft recipients were immunosuppressed with total lymphoid irradiation (TL1) and cyclosporine (CSA). After 100 days, islet function was assessed, at which stage immunosuppression was terminated. Glucose, insulin, glucagon, and C-peptide response was assessed during intravenous glucose tolerance test (IVGTT) and during arginine and tolbutamide stimulation. In eight normoglycaemic primates in which immunosuppressive treatment had been stopped and with mean graft survival of 145 days, islet stimulation was associated with moderate glucose intolerance, reduced K-values, hypoinsulinaemia, and low C-peptide values. Postmortem findings in all animals intentionally killed revealed severe graft atrophy in the absence of significant rejection. Severe graft atrophy in normoglycaemic primates, together with significantly impaired graft function after segmental pancreatic transplantation compared to normal animals, suggest that transplantation of the whole pancreas may be mandatory if normal or near-normal function is to be achieved.     

Special malignancy pattern in Chinese renal transplantation recipients: a single center experience and literature review

Malignancy is one of the main complications after renal transplantation but the situation in Chinese renal allograft recipients remains an enigma. We therefore reviewed 1,000 (8,531 person-years follow-up) renal allograft recipients from Jinling Hospital, Nanjing University, revealing an incidence rate of 2.4% of post-transplant malignancies, with a standardized incidence ratio (SIR) of 17.8 (95% C.I.: 16.7-18.8); the standardized rate is 1.67%, compared with 0.29% in the general population. However, our group demonstrated an extremely low incidence of skin cancer, which dominates in western countries. To confirm the findings, we reviewed the literature on post-transplant malignancies in Chinese renal allograft recipients, covering 296 malignancies in 18,548 renal transplant recipients in 21 reports. The top three most common sites of malignancies were the digestive tract, bladder and liver. The incidence of skin cancer was very low in most centers. These data show that Chinese renal allograft recipients have a unique spectrum of post-transplant malignancies, with an extremely very low incidence of skin cancer as compared to populations from western countries.     

Dose de tolérance à l’irradiation des tissus sains : larynx et pharynx

For head and neck cancers, the radiation dose usually needed to sterilize a macroscopic tumour is at least 70 Gy in conventional fractionation. In the larynx, this dose level enables optimal tumour control while exposing the patient to a limited risk of severe complications. For oropharynx and nasopharynx tumors, it is sometimes possible to limit the dose received by the larynx according to the extent of the primary lesion. Thus, if the tumour constraints permit, the maximum dose to the larynx must be less than 63 to 66 Gy. To reduce the risk of laryngeal edema, it is recommended if possible to limit the mean non-involved larynx dose to 40 to 45 Gy. In the pharynx, literature's data suggested to minimize the volume of the pharyngeal constrictor muscles receiving a dose greater than or equal to 60 Gy. Limiting the volume receiving a dose greater than or equal to 50 Gy reduces the risk of dysphagia. These dose constraints should be tailored to each patient taking into account the extent of the initial primary lesion, the possible addition of chemotherapy or a modified fractionation radiotherapy.