同种异体耳移植的模型建立

目的 建立同种异体耳移植的动物模型.方法 以青紫蓝兔和日本大耳白兔为实验动物.以耳大动脉、耳大静脉及外侧耳缘静脉为蒂,获得单侧(右)全耳,应用显微外科微血管吻合技术进行异体移植.其中青紫蓝兔为受体,日本大耳白兔为供体.实验共选用15对(青紫蓝兔和日本大耳白兔各15只)动物,随机分为两组:A组,对照组(n=5对),行自体兔耳离断回植术;B组,实验组(n=10对)行同种异体耳移植术.动、静脉吻合均选用端端吻合技术.术后观察移植耳存活、急性排异发生情况及供体组织的病理分级.结果 ①通过1%甲紫溶液灌注显示应用此方式获得的游离耳模型,在以耳大动脉、耳大静脉及同侧耳缘静脉为蒂供血后,可以保证耳移植后的存活.②A组自体移植复合组织瓣存活30 d以上.③B组异体移植复合组织瓣全部发生排斥反应.结论 建立的同种异体兔耳移植模型可重复性强,是进行相关研究的良好模型.     

同种异体耳移植的实验研究

目的：①观察同种异体兔耳移植后排斥反应的组织病理学、细胞免疫、体液免疫变化的情况;②为提高同种异体移植耳成活率,预防排斥反应,提供一个有效的免疫抑制治疗方案。方法：以青紫蓝兔和日本大耳白兔为实验动物。以耳大动、静脉及外侧耳缘静脉为蒂,获得单侧（右）全耳,应用显微外科微血管吻合技术进行异体移植。其中青紫蓝兔为受体,日本大耳白兔为供体。实验共选用40对（青紫蓝兔和日本大耳白兔各40只）动物,按受体免疫抑制剂治疗方案随机分为5组：O组,手术对照组（N=5对）,行自体兔耳离断回植术;C组,对照组（N=5对）,同种异体耳移植后不给予免疫抑制剂;T1组（N=10对）,同种异体耳移植后给予FK506单一免疫抑制剂;T2组（N=10对）,同种异体耳移植后给予FK506和强的松两联免疫抑制剂;T3组（N=10对）,同种异体耳移植后给予FK506、MMF和强的松三联免疫抑制剂。动脉、静脉吻合均选用端端吻合技术。术后观察移植耳存活情况、急性排异发生率及严重程度;供体组织病理分级;受体血浆IL-2、IL-4、IL-8的变化以及受体T淋巴细胞CD4^＋、CD8^＋的变化。结果：同种异体兔耳移植30天、100天后T3组在急性排斥反应发生率、供体组织病理分级和受体IL-2、IL-4、IL-8水平与CD4、CD8值的变化等方面与C组、T1组、T2组相比较均有显著差异（P〈0.05）。结论：三联免疫抑制剂方案（FK506、MMF和强的松）是预防同种异体兔耳移植排斥反应的一个有效治疗方案。     

犬头面部复合组织同种异体移植模型的建立

目的建立犬头面部复合组织同种异体移植的实验动物模型。方法①解剖研究：用4只杂种犬进行头面颈部的解剖及复合组织瓣的切取研究。②自体回植（Ⅰ组n=5）：为选择最佳的复合组织瓣移植方式，进行了三种形式的复合组织瓣白体回植。③异体移植（Ⅱ组n=6）：以第三种复合组织瓣及其改良的组织瓣形式进行了两种头面部复合组织异体移植。免疫抑制治疗方案为环孢霉素A和皮质激素联合应用，依据环孢霉素A血药浓度调整用药剂量。分别在术后4、6、12周及6个月对眼轮匝肌进行肌电图检测。结果①犬头面部的解剖特征与人相似，以颈外动静脉为血管蒂能够为头面部复合组织瓣提供良好的血供。②IC组两只犬长期存活，但出现唾液漏。③Ⅱa组中1只犬于移植术后28d免疫排斥，通过调整环孢霉素A及强的松的用量并局部应用氯倍他索涂抹，2周后治愈，现已存活309d。Ⅱb组中3只犬现已分别存活159、129和108d，无并发症，肌电图显示眼轮匝肌功能逐步恢复。结论改良犬单侧上半头面部组织瓣异体移植模型是研究头面部复合组织异体移植的理想模型。     

兔同种异体颜面复合组织移植后面神经再生的初步研究

目的 建立兔颜面部复合组织同种异体移植的实验动物模型,观察移植后面神经的再生情况.方法 选取日本大耳白兔16只,随机配对后分别作为供、受体,分为8对.建立兔同种异体颜面复合组织移植的实验动物模型,FK506与皮质激素联合应用作为免疫抑制治疗方案.术后14、28、40d取移植面神经吻合口处标本分别行HE染色光镜和透射电镜观察,术后28、40d分别行神经肌电图检测,分析移植后兔面神经的再生情况.结果 8例移植手术中,6例获得成功.移植后14d神经纤维以变性改变为主,有少量神经纤维长入吻合口,28d移植神经的吻合口有较多的神经纤维长入,神经束排列较前规则,40d再生神经纤维变得较前成熟,有髓神经的数量增多,但仍可见到变性的神经纤维.术后28d未测得明显的神经动作电位,40d测得的神经动作电位波形离散,表现为明显的传导阻滞.结论 同种异体颜面复合组织移植后面神经的再生是个极为缓慢的过程,需要经历一系列的病理生理过程.移植后40d神经吻合口可出现组织结构上较为成熟的再生神经纤维,但要达到功能上的恢复尚需更长时间.     

自体静脉旁路移植术后再狭窄的实验研究

目的 观察自体静脉旁路移植后主体动脉段的血管壁的变化.方法 新西兰大白兔18只,取右侧颈外静脉旁路移植至同侧颈总动脉为实验组,仅游离左侧颈总动脉为假实验组,正常左侧颈总动脉为正常对照组,术后6周取材光镜下观察、测量,比较各血管段内膜、中膜变化差异,电镜观察细胞形态变化.结果 游离的动脉与正常动脉相比无显著性差异(P>0.05);主体动脉近远端底部及盲段内膜明显增厚(P<0.01).结论 自体静脉旁路移植术后,主体动脉近远端底部及盲段内膜明显增厚,考虑与局部血流动力学改变密切相关.     

国外美容医学最新动态

1灵长类动物面部复合组织同种异体移植1：延长移植体存活的技术及免疫抑制疗法 目的：由于排斥反应及传统的免疫抑制剂的应用带来的风险,阻碍了复合组织同种异体移植手术的广泛开展。本文作者为了给临床应用提供更多的实验资料,用灵长类动物进行面部复合组织同种异体研究,并观察其长期的效果。方法：在匹配不良的猕猴间,施行面部复合组织（包括皮肤、肌肉、骨骼）异体移植实验。从供体切取面部复合组织,保留供血的颈总动脉,和颈内、颈外两条引流静脉。将移植体转移至另一猕猴的下腹部-异位移植。供体的颈总动脉与受体的股动脉进行端-端吻合,两条颈静脉分别与股静脉吻合。受体动物接受单一的tacrolimus免疫抑制剂治疗。术后不间断地观察,适时取活体组织检查或直行核磁共振检查。结果：在13个移植体中,两例于术后3～5天因一条静脉栓塞而失效,与排斥无关,其余移植体均成活良好。单一的免疫抑制疗法有效,未发生并发症。移植体的无排斥反应存活期为60～177天。结论：作者利用灵长类动物猕猴施行面部复合组织同种异体移植获得成功。保持两条引流静脉的畅通是成功的要点,采用tocrolimus单项免疫抑制剂可获长期的抑制排斥反应的效果,未出现早期并发症。此模型可作为进一步研究复合组织同种异体移植的研究工具。     

兔同种异体异位膝关节移植模型的建立

目的建立兔同种异体异位膝关节移植实验动物模型.方法在观察兔的下肢解剖的基础上,设计兔同种异体异位膝关节移植术;观察吻合血管同种异体膝关节移植组的自然病程.结果兔吻合血管同种异体膝关节移植共9例,移植成功7例,移植物平均存活(13.57±0.79)d,移植物液化坏死,受体生命体征受排斥反应影响大.结论兔同种异体异位膝关节移植模型操作简便,表现稳定,适用于复合组织移植免疫的研究.     

近交系大鼠腹部游离皮瓣移植模型的建立及意义

目的:为异体复合组织移植及皮肤移植免疫学研究建立一种可靠、简便而实用的动物模型。方法:以近交系Brown Norway(RT1n)大鼠为供体,Lewis(RT11)大鼠为受体,以股动静脉为血管蒂,进行腹部游离皮瓣异体移植。实验分为2组:急性排斥组(A组)术后不予免疫抑制治疗;免疫抑制组(B组)术后腹腔注射CsA剂量为20mg/kg/天。术后观察移植皮瓣的排斥情况和皮瓣存活时间,并进行组织病理学检查,以此对动物模型的可靠性和实用性进行评价。结果:A组移植皮瓣均在术后(7±1)天发生明显的逐渐加重的急性免疫排斥反应;B组移植皮瓣均可良好存活,存活时间均>100天。组织病理学检查,A组皮瓣有血管炎、毛囊炎及真皮炎症等典型急性排斥反应表现;B组皮瓣均无明显免疫排斥病理学表现。术后IL-2水平与皮瓣表面及病理学检查所见排斥反应程度相一致。结论:近交系大鼠腹部游离皮瓣移植模型是操作更为简易的血管化皮肤移植模型,其免疫排斥的发生易于观察并可稳定控制,是异体皮肤移植免疫学研究的良好动物模型。     

大鼠同种异体喉移植模型的建立

应用现代显微外科手术，将双侧连接于体供喉的颈总动脉分别与受体一侧的颈总动脉和颈外静脉进行端－侧吻合，共对１３只大鼠实施喉异位移植。建立了同种异体喉移植的实验模型。移植动物全部存活，移植术后３天，移植物周围有纤维素粘粘连，７天后粘连加重，移植物表面附有灰白色的纤维结缔组织气管内     

近交系大鼠后肢移植模型的建立及意义

目的:介绍异体复合组织移植研究中操作难度较大的大鼠后肢移植模型的建立方法,操作体会及此动物模型在复合组织移植中的意义。方法:以近交系BrownNorway(RT1n)大鼠为供体,Lewis(RT11)大鼠为受体,吻合股动、静脉为供血血管,进行近交系大鼠后肢异体移植。实验分为2组:急性排斥组(A组)术后不予免疫抑制治疗;免疫抑制组(B组)术后腹腔注射FK506剂量为1mg/kg/天。术后观察移植肢体的排斥情况及其存活时间,并取皮肤进行组织病理学检查,以此对动物模型的可靠性和实用性进行评价。结果:A组移植皮瓣均在术后(4±1)天发生明显的逐渐加重的急性免疫排斥反应;B组移植肢体均可良好存活,存活时间均>100天。皮肤组织病理学检查上,A组有血管炎、毛囊炎及真皮炎症等典型急性排斥反应表现;B组均无明显免疫排斥病理学表现。术后IL-2水平与肢体外观表现及病理学检查所见排斥反应程度相一致。结论:近交系大鼠后肢移植模型是得到广泛承认的复合组织移植模型,也是最具挑战性的动物模型之一,完善近交系大鼠后肢移植模型的建立方法,使模型不断规范化,对复合组织异体移植研究的发展具有重要意义。     

异体复合组织移植皮肤淋巴引流隔绝动物模型的建立

目的:通过SD大鼠下腹部游离皮瓣及皮管的移植,建立大鼠异体复合组织移植皮肤淋巴引流隔绝模型。方法:7～8周龄雄性SD大鼠切取下腹部皮瓣并游离行同种异体移植,皮肤隔离组(A组):将供体皮瓣边缘对合、缝合成管状,底边留血管蒂出口用隔离器隔离皮肤,显微外科吻合血管,皮管缝合于隔离器,隔离器固定于受体;皮肤未隔离组(B组):将供体游离皮瓣卷制成管状,显微外科吻合血管并将供受体皮肤对位缝合;空白对照组(C组):行下腹部游离皮瓣异体移植。大体观察每组皮瓣或皮管存活时间,于术后3、5、7、14、28、35d取移植物皮肤组织行HE染色,观察病理学变化。结果:A组:皮管存活时间为32(34,32)天;B组:皮管存活时间为15(16,15)天;C组:皮瓣存活时间为6(7,6)天。A组与C组、B组与C组、A组与B组存活率差异均有统计学意义(P<0.05)。结论:本实验成功建立了大鼠的异体复合组织移植淋巴引流隔绝模型,并证实了隔绝皮肤淋巴引流能有效延长移植物的存活时间。     

Experimental Canine Facial Transplantation

Objective

Facial allotransplantation represents a novel frontier in the reconstruction of complex human facial defects. To develop more refined surgical techniques and yield fine results, it is required to make a suitable animal model. The development of a model of composite facial and scalp allograft in canines is more appealing: In large animals, canine facial anatomy is the most similar to humans; its facial nerve anatomy also resembles humans'; and canines possess the most similar facial vascular anatomy to humans. These factors led to the development of a canine composite facial allograft model.

Methods

Two operative teams performed simultaneously on both donor and recipient to harvest the hemifacial/scalp flap and to prepare the recipient to shorten operative time. The flap was harvested with the common carotid artery and external jugular vein and transferred to the recipient. After insetting of the cartilage, skin, and muscles, the anastomosis of the external jugular vein and anastomosis between the external carotid artery and lingual artery were performed.

Results

The total mean time of transplantation was 5 hours ± 30 minutes. All of the transplanted animals were wreated with FK-506 [tacrolimus, 2 mg/kg] for 7 days after surgery. Clinical rejection response was also identified by close monitoring. Most allografts survived with perfect viability without vascular problems in the early postoperative period.

Conclusions

We documented that this model is well qualified in every aspect for use as a standard transplantation training model and future research work.     

Laryngeal allotransplantation by stages in beagles: an initial study

Objective

To establish a model of cynolaryngeal transplantation.

Methods

Ten beagles, matched for weight (8–10 kg) were divided into five groups for allotransplantation. The vascular circulation was managed with an end-to-end or end-to-lateral anastomosis of allograft bilateral common carotid arteries to recipient bilateral common carotid arteries, and bilateral external jugular veins. The recurrent laryngeal nerve was anastomosed end-to-end. Then, a vascularized allograft was embeded in the cervical subcutaneous tissue of the recipient, and cyclosporine administered every day.

Results

The allografts in groups 3, 4, and 5 survived up to 1 month. Groups 1 and 2 showed necrosis and cervical infection.

Conclusions

Beagles accepted laryngeal allotransplantation by stages (laryngeal allograft fore-embed model). Allograft survival was secluded by glandular organ secretion. Immunosuppression must be continuously administered.     

深低温冷冻技术对带血管同种异体骨移植抗原性作用的实验研究

[目的]探讨深低温冷冻技术对带血管同种异体骨移植物抗原性的影响,观察移植骨的吻合血管通畅率和骨连接情况,了解免疫抑制剂的用量。[方法]健康成年新西兰白兔48只,随机取16只作供体,32只作受体,分为5组。A组:8只,深低温冷存的带肱血管的异体桡骨段移植,手术后4周加用CsA(每天10 mg/kg);B组:8只,深低温冷存的带肱血管的异体桡骨段移植,手术后4周加用CsA(每天5 mg/kg);C组:8只,深低温冷存的带肱血管的异体桡骨段移植,手术后4周加用CsA(每天2 mg/kg);D组:8只,深低温冷存的带肱血管的异体桡骨段移植;E组:16只供体兔取移植骨段后继续喂养4周,作为免疫学检测,无移植对照组。术后观察吻合血管通畅率,术后7 d血清IFN-γ水平检测,术后2、4、8、12周各组动物X线检测,术后12周移植骨组织学检查。[结果]A、B组之间血清IFN-γ水平无显著差异,A、B组血清IFN-γ水平低于C组。[结论]深低温冷冻技术降低了带血管同种异体骨移植物的抗原性,移植后显著降低器官的移植排斥反应,保证吻合血管的通畅,移植骨的存活,加快骨连接,减少免疫抑制剂的用量。     

Microvascular reconstruction of the pharyngoesophagus with free jejunal graft

Free jejunal graft reconstruction of the pharyngoesophagus has become reliable one-stage technique, but the microsurgeon must be aware of numerous pitfalls. Forty-two patients were retrospectively reviewed. There were four graft failures for a success rate of 90.5%. When harvesting the graft the thick fatty mesentery may make dissection difficult, and injuries to the mesenteric artery and vein can occur. Neck preparation can be tedious because of previous radiation, and size discrepancies with the carotid branches and the mesenteric artery are seen. Intimal dissection of the mesenteric artery can present as an isolated flap or a circumferential "rosette," despite the most careful preparation of the vessels; and great care must be taken in the microvascular anastomosis. The end-to-end arterial anastomosis to a branch of the external carotid and an end-to-side venous anastomosis to the internal jugular were preferred. Vein grafts are occasionally necessary.     

Free flaps for head and neck cancer reconstruction: Does the use of both large cervical vessels as recipient vessels and the employment of end-to-side technique enhance flap survival?

This study specifically investigates whether the use of both large cervical vessels (the external carotid artery and the internal jugular vein) as recipient vessels with end-to-side anastomosis enhance free flap survival in head and neck cancer reconstruction, when compared with the use of other standard smaller neck recipient vessels and end-to-end anastomosis. A total of 84 consecutive patients were included and were divided into two groups (42 in each group) according to the recipient vessels. The overall vessel thrombosis rate was 6% (five of 84 cases) and the overall flap loss rate was 2.4% (two of 84 cases) yielding a flap salvage rate of 60%. Vessel thrombosis occurred in three cases of the smaller vessels group and in two cases of the large cervical vessels group. This was not statistically significant.     

Versatility of a posterior-wall-first anastomotic technique using a short-thread double-needle microsuture for atherosclerotic arterial anastomosis

To perform safe anastomosis for atherosclerotic arteries, we employed a posterior-wall-first anastomotic technique with a short-thread double-needle microsuture. Between October 2005 and September 2007, we performed 17 free flap transfers to the head and neck regions with this technique. Fifteen of 17 patients had one or more risk factors for atherosclerosis. In all patients except one, the superior thyroid artery was employed as a recipient artery and end-to-end anastomosis was performed close to its origin with the external carotid artery. Postoperatively, no anastomotic thrombosis occurred in any patients. The greatest advantage of the double-needle microsuture is that each stitch can be placed from the inner side of the arterial wall to the outer side. This "inner-to-outer" stitch secures full-thickness bite and minimizes intimal damage. Safe and reliable anastomosis could be carried out in an unfavorable setting because the double-needle suture offers a high degree of freedom in suture placement.     

Technical aspects of segmental pancreatic autotransplantation in dogs

Heterotopic segmental pancreatic autotransplantation in dogs is more appropriate than allograft models for the investigation of several problems associated with transplantation. We have defined the anatomic variations of blood supply in the pancreatic tail and designed various modifications of vascular anastomosis to the iliac vessels in order to eliminate technical failures, such as thrombosis, as much as possible. In 187 of 240 dogs (77.9 percent), the pancreatic artery originated from the splenic artery and the pancreatic vein entered the splenic vein (normal anatomy). The main venous variation was direct confluence of the pancreatic and portal veins (12.1 percent), and the main arterial variation was origin of the pancreatic artery from the superior mesenteric artery (10 percent). Ninety-seven animals with normal anatomy qualified for a comparative study of seven methods of segmental pancreatic autotransplantation. Venous anastomoses were always performed in an end-to-side fashion between the splenic and external iliac veins. Arterial anastomosis techniques follow. Group I: interposition of the splenic artery into the external iliac artery; (14 days, failure rate 50 percent), Group II: end-to-end arterial anastomosis of the splenic artery to a long external iliac artery segment with the graft directed caudad, resulting in an acute curve to the vessel loop (8 dogs, failure rate 38 percent); Group III: end-to-end arterial anastomosis to a long external iliac artery with the graft directed cephalad, resulting in a gentle curve to the vessel loop (11 dogs, failure rate 36 percent); Group IV: end-to-end arterial anastomosis to a short external iliac artery stump (20 dogs, failure rate 20 percent); Group V: same as in Group IV with the addition of a distal splenic arteriovenous fistula (12 dogs, failure rate 17 percent); Group VI: end-to-side anastomosis of the splenic artery to the external iliac artery (31 dogs, failure rate 6 percent); Group VII: same as in Group VI but with the addition of an arteriovenous fistula of the distal splenic vessels (1 dog, failure rate 0). The end-to-side technique proved to be straight-forward and reliable. The low failure rate with this method allows metabolic preservation and other aspects of pancreatic transplantation to be studied and the results to be interpreted without the influence of a high complication rate from the operation itself.     

Comparison of continuous and interrupted suture techniques in microvascular anastomosis.

We evaluated the efficacy of the continuous suture technique (CST) in arteries and veins with varying external diameters (ED). In study 1 a direct end-to-end anastomosis was performed in 5 groups of animals (n = 15 in each group): group 1, rabbit carotid artery (ED, 1.8-2.0 mm); group 2, rabbit femoral artery (ED, 1.4-1.6 mm); group 3, rat femoral artery (ED, 0.7-0.9 mm); group 4, rabbit femoral vein (ED, 2.0-2.2 mm); and group 5, rat femoral vein (ED, 1.0-1.2 mm). In study 2 a graft from the femoral vein was interposed into the carotid artery, with a ratio of the diameter of graft to artery of 1.3:1 in the rats (group 6, n = 12) and 1:1 in the rabbits (group 7, n = 12). In each animal the vessel on one side was repaired using CST and the opposite vessel using the interrupted suture technique. Vessel samples were harvested 1, 2, and 4 weeks after anastomosis. The CST significantly reduced anastomosis time by up to 47% in arteries and 41% in veins. Bleeding time and blood loss were also significantly reduced with CST. Similar results were found in study 2. The total thrombosis rate was 8%, but no significant patency difference was noted between the CST and the interrupted suture technique in any vessel category. We conclude that the CST is a reliable and time-saving procedure in microvascular anastomosis of arteries with diameters greater than 0.7 mm and of veins with diameters greater than 1.0 mm.     

猕猴异体子宫移植技术的应用研究

目的探讨猕猴异体子宫移植活体捐赠手术技巧应用,为下一步人体子宫移植临床试验提供实验数据。方法选用性成熟、月经规律的雌性猕猴2只,根据血型和体重选取供体1例和受体1例,将切除的供体子宫及髂血管给予器官保存液冷保存、灌注后原位移植到受体盆腔,供体的髂内动脉、髂外静脉与受体的髂外动、静脉使用端-侧吻合法间断吻合,术中根据移植血管的搏动、子宫的色泽判断移植是否成功,术后给予他克莫司、环孢霉素、泼尼松三联免疫抑制剂预防移植免疫排斥。结果受体猕猴移植双侧髂血管吻合成功,可触及移植血管搏动,移植子宫恢复血流灌注后子宫颜色由灰白逐渐转为暗红。受体动物于子宫移植术后28d死亡,评估死亡原因可能为慢性衰竭。结论猕猴是适用于异体子宫移植研究的大型动物模型,使用髂内动脉与受体的髂外动脉进行端-侧吻合法吻合是可行的,三联免疫抑制剂可有效预防移植免疫排斥。