小鼠异位肢体复合组织移植模型的建立

目的建立稳定的小鼠异位肢体复合组织移植模型。方法选用25对C57BL/6小鼠行同品系异位肢体复合组织移植手术。取供体小鼠后肢,于腹股沟韧带下缘起沿股动脉、股静脉向上游离,电凝切断除股静脉、股动脉外所有分枝血管至髂外动脉、髂外静脉,断髂外动脉、髂外静脉后,横断股骨、股部肌肉。将供体髂外动脉、髂外静脉分别与受体小鼠右侧颈总动脉、颈外静脉行端侧吻合。连续缝合供体皮缘于受体皮肤切缘固定肢体。于供肢胫骨远端关节处切断,结扎残端。结果手术成功率76%,总手术时间(87.1±10.1)min,供体获取时间(23.8±4.9)min,血管吻合时间(55.3±9.6)min。结论小鼠异位肢体复合组织移植成功率较高,稳定可靠,是一种新的研究移植免疫的理想的动物模型。     

动脉不同剪切力实验动物模型的建立

目的寻找一种简单、准确的方法建立动脉的不同剪切力动物模型,为研究剪切力在心血管系统中的作用及机制奠定基础。方法 30只SD大鼠随机分为3组,即颈总动脉结扎组、股动静脉吻合组和髂总动脉结扎组,每组又分为手术组和假手术组(对照组),利用多普勒超声仪检测手术前后动脉血流,估算血流剪切力的变化。利用免疫荧光组织化学法检测不同剪切力下血管内皮e NOS的表达情况。结果颈总动脉结扎法大鼠死亡率高;股动静脉吻合法手术耗时长,且因动脉太小不易测得股动脉血流;单侧髂总动脉结扎手术可以使双侧髂总动脉分别形成不同血流剪切力。单侧髂总动脉结扎术后,结扎侧髂总动脉内皮e NOS免疫反应性减弱,而结扎对侧e NOS免疫反应性增强。结论单侧髂总动脉结扎手术简单易操作,髂总动脉血流速度通过多普勒超声仪可获得,是制作不同剪切力动物模型的最佳方法。     

吻合皮下静脉的带蒂皮瓣修复四肢皮肤软组织缺损的效果分析

摘要 目的：探讨与分析吻合皮下静脉的带蒂皮瓣修复四肢皮肤软组织缺损的效果。方法：选择2018年12月到2021年12月在本院创伤造成的四肢皮肤软组织缺损60例患者作为研究对象,将其随机分为吻合皮下静脉带蒂皮瓣组与传统带蒂皮瓣组各30例。吻合皮下静脉带蒂皮瓣组给予吻合皮下静脉的带蒂皮瓣修复治疗,传统带蒂皮瓣组给予常规直接覆盖创面修复治疗。结果：所有患者都顺利完成手术,吻合皮下静脉带蒂皮瓣组围手术指标时间均较传统带蒂皮瓣组少（P<0.05）。吻合皮下静脉带蒂皮瓣组术后3个月的总有效率为96.7 %,高于传统带蒂皮瓣组的76.7 %（P<0.05）。吻合皮下静脉带蒂皮瓣组术后3个月的并发症发生率较传统带蒂皮瓣组低（P<0.05）。吻合皮下静脉带蒂皮瓣组术后6个月的感觉功能恢复情况好于传统带蒂皮瓣组（P<0.05）。结论：吻合皮下静脉的带蒂皮瓣能促进患者的创面愈合,提高治疗效果,减少并发症,加快恢复患者的四肢皮肤软组织缺损。     

部分股骨移植:一个新的带血管的骨髓移植模型

目的:大鼠后肢移植是带血管的骨髓移植的主要模型,同时含有骨髓和骨髓微环境,有利于免疫耐受的诱导.但其成分复杂,为研究其中骨髓的作用,构建一个成分相对单一的带血管的骨髓移植模型.方法:以MHC不相符的近交系Lewis和BN大鼠分别作为供体或受体,将含血管蒂的供体2/3股骨移植到受体腹股沟区,显微吻合供受体间的股动静脉.实验分为3组:同基因移植组Lewis→Lewis;排斥组Lewis→BN;免疫抑制组Lewis→BN,术后给予环孢素A.通过大体和病理学检查观察各组移植物存活情况,外周血流式监测排斥组和免疫抑制组的嵌合水平.结果:术后7d,排斥组排斥反应显著,骨髓细胞明显减少、坏死,外周血嵌合水平几乎为0,而同基因移植组股骨存活良好,骨髓HE染色大致正常,可见髓腔内有大量的嗜碱性骨髓细胞;术后60 d,同系移植组和免疫抑制组的大体观察及组织病理学大致相同,均证实移植物存活良好,同时流式可见免疫抑制组术后7、28、60 d外周血中存在供体特异性的嵌合.结论:带血管蒂的部分股骨移植物是一个简便可靠的带血管的骨髓移植模型,嵌合分析表明其具有诱导耐受的潜能,有利于阐明异体复合组织中骨髓对免疫耐受诱导的作用和机制.     

膝内侧游离皮瓣修复足跟软组织缺损的研究与应用

目的:探讨膝内侧游离皮瓣修复足跟皮肤软组织缺损的临床效果.方法:2007-2009应用膝内侧游离皮瓣移植修复足跟皮肤软组织缺损12例.将膝降动脉和其伴行静脉与胫后动静脉吻合,皮瓣大隐静脉与受区大隐静脉吻合,将股内侧皮神经、隐神经与受区腓肠神经或隐神经吻合,使皮瓣具有良好的血运和感觉功能;将缝匠肌腱和股薄肌腱重建跟腱缺损修复跟腱功能.结果:移植皮瓣全部成活,创面一期愈合,经6周-18个月随诊,皮瓣感觉恢复良好,足跟外形及稳定性良好.结论:膝内侧游离皮瓣血管口径粗,蒂长,带有两根知名感觉神经,皮瓣较薄,修复足跟外形美观,耐磨,稳定好,特别是带有缝匠肌腱和股薄肌腱修复跟腱缺损,是足跟部皮肤软组织缺损修复良好供区.     

小型猪胰腺移植急性排斥反应模型的实验研究

目的　建立小型猪胰腺移植动物模型 ,探索早期诊断急性排斥反应的方法。方法　 4 0只猪随机配对行胰腺移植 2 0次 ,将供胰所带的血管与受体髂血管吻合 ,所带小段十二指肠与空肠吻合 ,术中监测平均动脉压、中心静脉压及血气。术后测定受体的血淀粉酶、血糖 ,监测外周血免疫指标 ,彩色多普勒检测供胰血流及超声引导下活检、组织病理检查。结果　移植手术成功率为 90 % ,受体平均动脉压在吻合血管开放后有明显下降 ,与血流开放前差异有显著性 (P <0 0 5 ) ,输血有助于手术成功 ,受体术后平均存活 (12 6± 2 3)d。外周血免疫学监测指标早于供胰的组织病理改变 ,两者的改变均早于急性排斥反应的临床表现。结论　小型猪适用于胰腺移植模型的建立 ,加强术中循环功能的管理、及时输血有利于受体成活 ;超声引导下穿刺活检供胰的组织病理学检查与监测外周血免疫指标均适于早期诊断急性排斥反应     

大鼠腹腔异位心脏移植模型建立及改进

目的:建立一种快速有效的大鼠腹腔异位心脏移植模型。方法:采用SD大鼠作为受体,Wistar大鼠作为供体,行同种异位腹腔心脏移植,术后给以CsA5 mg/kg/d灌服,心脏移植手术方法采用改良的Ono术式,观察改良的腹腔异位心脏移植各步骤所需时间、术后成功率及主要并发症发生率。结果:共建立40只大鼠异位心脏移植模型,手术成功率92.5%。动脉吻合时间12.5±2.3min,静脉吻合时间12.3±1.5 min,供心缺血时间37±3.5 min,受体血管阻断时间34.2±2.6 min,总手术时间90.2±4.8 min,出现的主要并发症为出血和供心复跳失败(各占5%、2.5%)。结论:改进的大鼠腹部异位心脏移植技术是一种简便、快速、有效、成功率高的模型制作方法。     

小鼠Lewis肺癌不同部位皮下移植瘤模型的比较

目的探究不同部位肺癌皮下移植瘤存在的差异,为肺癌研究者提供提供基础依据。方法将稳定表达虫萤光素酶的小鼠Lewis肺腺癌细胞(ll2-luc-m38)分别注射于C57 BL/6小鼠的右腋皮下、右腹股沟皮下、脚垫皮下,定期利用小动物活体成像系统观察小鼠皮下移植瘤成瘤情况及转移情况,观察不同部位荷瘤小鼠的生存时间和死亡率,肿瘤组织取材,采用石蜡包埋、切片、HE染色做出病理学诊断。并将小鼠处死后立即进行肺部组织固定、观察转移灶。对皮下移植瘤行切除术观察小鼠术后生存情况和转移情况。结果腋下组和腹股沟组成瘤时间较早,成瘤率为100%,脚垫组成瘤时间较晚,成瘤率为33%;腹股沟组和腋下组瘤体积增长迅速,其中腹股沟组瘤体积增长最快;接种后第21天腋下组70%的小鼠出现了肺转移,转移灶数目较多;腹股沟组50%小鼠出现肺转移,且转移灶数目较少;脚垫组小鼠未观察到肺转移灶。脚垫组小鼠死亡率最高。腹股沟组和腋下组小鼠可行皮下移植瘤切除术,术后存活率为100%。结论小鼠lewis肺癌皮下移植瘤模型中,腹股沟组和腋下组成瘤率高,可耐受移植瘤切除术,手术死亡风险低,便于监测,操作简单且可重复性高,其中腋下组转移性更好。     

负载供者抗原的第三方树突状细胞延长大鼠移植心脏存活

目的观察负载供者抗原第三方未成熟DC对大鼠同种异体移植心脏存活期的影响。方法培养骨髓来源第三方大鼠未成熟DC,负载供者抗原,CTLA-4 Ig致耐受处理。建立大鼠心脏腹部移植模型,A组：心脏移植;B组：心脏移植,术前输注供者源未成熟DC;C组：心脏移植,术前输注第三方未成熟DC;D组：心脏移植,术前输注负载供者抗原致耐受第三方未成熟DC。分别行移植心病理、IFNγmRNA表达和移植心存活时间观察。结果D组的移植心病理改变轻,IFNγmRNA表达下调,移植心脏存活时间显著延长。结论负载供者抗原第三方致耐受未成熟DC能够诱导供者特异性免疫耐受,明显延长大鼠移植心脏存活时间。     

大鼠心脏移植急性排斥反应动物模型的建立

目的为探索器官移植后急性排斥反应的一种快速可靠的诊断方法。方法先暴露和游离受体的吻合段血管,再获取供心以缩短移植心脏冷缺血时间;保留供者的心脏及右上肺,将其胸主动脉与受者的腹主动脉间断端侧吻合;开放血流时,先开放远心端再间断开放近心端,同时按摩心脏至心脏搏动规律、有力,其色泽恢复红润,右上肺亦充盈良好。结果A组(n=20)成功16只,动脉吻合口出血2只、心脏复跳失败1只、吻合口狭窄1只;B组(n=20)成功17只,2只死于吻合口出血,1只死于吻合口狭窄。A组手术成功率为80%,B组为85%(P>0.05)。结论(1)先暴露并游离好受体血管吻合段,再获取供心,可以缩短移植心冷缺血时间。(2)边灌注边获取的方法利于快速而又完整的获取供心。(3)在切取和植入的过程中一定要注意低温保护。(4)利用腹主动脉段进行移植,使手术方便易行。左心做功的大鼠腹部心脏移植模型简单、安全、实用,因其左心参与循环而使血流动力学更接近生理状态,是进行心脏移植及血流动力学研究的较理想模型。     

A new flap design: neural-island flap

This report introduces the "neural-island flap" concept, which represents a consistent and reliable skin flap design supplied only by the intrinsic vasculature of a cutaneous nerve. In this study, the lateral femoral cutaneous nerve was selected as the pedicle of the neural-island flap, and a standard skin flap, which is the territory of the accompanying vessels (i.e., iliac branches of the iliolumbar artery and vein), was elevated on the lower dorsal region of the rats. In a total of 92 Wistar rats, three experiments were performed. In part I (n = 24), the vascular anatomy of the lateral femoral cutaneous nerve was established by the methods of dissection, microangiography, nerve mapping, perfusion with colored latex and India ink, and histologic analysis. In part II (n = 46), the role of the cutaneous nerve in supporting an acutely elevated skin flap was explored by creating five flap groups as follows: group 1, conventional flap (artery, vein, and nerve intact); group 2, neural island flap (only the nerve intact); group 3, neurocutaneous flap (vein and nerve intact); group 4, denervated flap (artery and vein intact); and group 5, skin graft. In part III (n = 22), the role of a preliminary surgical delay procedure to augment the survival of the neural island flap was investigated. Results of the anatomic studies indicated a consistent perineural vasculature by the accompanying iliolumbar artery. Skin flaps survived totally in groups where the artery and vein were intact, whereas mean survival rates for the neural island flap and the neurocutaneous flap were 38.2 +/- 3.1 percent and 44.5 +/- 3.8 percent, respectively (p > 0.05). Results of part III of the experiment demonstrated a significantly higher survival for the delayed neural island flap (94.5 +/- 5.5 percent) compared with the acutely elevated neural island flap (p < 0.05). The perineural and intraneural vessels were found to be greatly dilated after a delay procedure, demonstrated by direct observation, microangiography, histologic analysis, dye injection study, and scanning electron microscopy. On the basis of this promising series of experiments, a clinical technique was developed using the sural neural-island flap. The flap was used to reconstruct lower extremity defects in four cases. A delay procedure was accomplished in the first stage by elevating a fasciocutaneous flap from the midcalf region based on a posterior skin bridge and the sural nerve. After a 2-week delay period, a sural neural-island flap was created based on the nerve and transposed to the defect. Flap survival was complete in all cases, with a satisfactory result. The authors conclude that this report proves for the first time that a robust and reliable skin flap can be created pedicled only by the intrinsic vasculature of a cutaneous nerve, after a proper surgical delay. The so-created neural-island flap design offers two novel advantages: (1) a very narrow pedicle and (2) a pedicle without any restriction to a specific pivot point, in addition to the previously described unique advantages of preservation of a major artery and avoidance of microvascular anastomoses.     

硬膜外导管在大鼠肾移植血管吻合中的应用

目的建立一种手术难度低,成功率高的大鼠肾移植模型。方法 Wistar大鼠作供体,SD大鼠作受体,将供体腔静脉与受体肾静脉端端吻合,供体腹主动脉与受体腹主动脉端侧吻合,供体膀胱瓣与受体膀胱吻合。根据血管吻合时应用硬膜外导管与否,将受体分为有支架组和无支架组两组。结果有支架组共进行肾移植30次,成活26只;无支架组共进行肾移植20次,成活10只。有支架组的成活率86.7%（26/30）较无支架组50.0%（10/20）明显提高（P〈0.05）,血管吻合总时间（22±2）min较无支架组（32±2）min明显缩短（P〈0.05）。结论硬膜外导管应用于大鼠肾移植血管吻合,降低了手术难度,减少了吻合口出血,提高了手术成功率。     

Surgical technique in the rat model of kidney transplantation

Today successful kidney transplantation procedures, techniques and immunosuppression protocols are a consequence of extensive research on animal models. During every transplantation surgery there are two crucial points for the success of the entire procedure: vascular (arterial end venous) and ureteral or ureterovesical anastomosis. Renal artery and vein of the donor kidney can be anastomosed end-to-side to the abdominal aorta and vena cava of the recipient (heterotopic transplantation), or end-to-end to the remains of renal artery and vain of the recipient (orthotopic transplantation) after nephrectomy. The ureter can be anastomosed also end-to-end or we can connect it directly to the urinary bladder (ureterocystoneostomy). The aim of this study was to elucidate which technique has better results according to: animal survival, reperfusion and perfusion of the transplanted kidney, elimination of the urine from the transplanted kidney and procedure costs. The study included 240 (120 donors and 120 recipients) male Wistar rats (3 months old; weight 250-300 g Our results are clearly showing that the end-to-end vascular anastomosis, and Paquins ureterovesical anastomosis have better results in transplanted rat kidneys survival and urine drainage compared to end-to-side vascular anastomosis and end-to-end ureteral anastomosis. Based on our experience we can conclude that described methods of end-to-end vascular anastomosis and Paquins ureterovesical anastomosis are less technically demanding and have a shorter learning curve. Therefore, we can recommend the use of described methods in kidney transplantation related researches.     

Application of the cross-bridge microvascular anastomosis when no recipient vessels are available for anastomosis: 85 cases

The purpose of this article is to introduce the results of free tissue transfers using the technique of the cross-bridge microvascular anastomosis when the recipient lacks suitable vessels for anastomosis. Between May of 1982 and June of 2002, a series of 85 patients underwent this procedure. The transferred tissues were the free latissimus dorsi myocutaneous flap, the free vascularized fibula, the free fibular osteocutaneous flap, and the free iliac osteocutaneous flap, alone or in combination. The donor vessels were the anterior tibial artery and great saphenous vein, the posterior tibial artery and its venae comitantes, and the radial artery and cephalic vein. Good results were achieved. The success rate reached 95.29 percent. The authors believe this procedure can be performed in the event of serious tissue defect where the vessels are unsuitable for anastomosis.     

Bone Marrow Transplantation Concurrently Reconstitutes Donor Liver and Immune System across Host Species Barrier in Mice

Liver immunopathologic mechanisms during hepatotropic infection, malignant transformation, and autoimmunity are still unclear. Establishing a chimeric mouse with a reconstituted liver and immune system derived from a single donor across species is critical to study regional donor immune responses in recipient liver. Using a strain of mice deficient in tyrosine catabolic enzyme fumarylacetoacetate hydrolase (fah ^-/-) and bone marrow transplantation (BMT), we reconstituted the donor''s hepatocytes and immune cells across host species barrier. Syngeneic, allogeneic or even xenogeneic rat BMT rescued most recipient fah^-/- mice against liver failure by donor BM-derived FAH⁺ hepatocytes. Importantly, immune system developed normally in chimeras, and the immune cells together with organ architecture were intact and functional. Thus, donor BM can across host species barrier and concurrently reconstitutes MHC-identical response between immune cells and hepatocytes, giving rise to a new simple and convenient small animal model to study donor''s liver immune response in mice.     

Pregnancy established in an infertile patient after transfer of a donated embryo fertilised in vitro.

In vitro fertilisation after stimulation of the ovulatory cycle has led to successful pregnancy. If more oocytes are recovered than are needed they may be left unfertilised, preserved, or donated to a recipient couple from whom oocytes cannot be obtained. A case of human pregnancy initiated by transfer of a donated embryo fertilised in vitro is reported. The donor was a 42 year old woman with primary infertility from whom six follicles were aspirated after stimulation of the ovulatory cycle. The recipient was a 38 year old infertile woman who had undergone several unsuccessful attempts for artificial insemination from a donor. Five oocytes were recovered from the donor''s six follicles, four of which were inseminated with spermatozoa of the donor''s husband and the fifth with a frozen sample of semen. Three of the four embryos fertilised by her husband were returned to the donor and the fifth was transferred to the recipient. No pregnancy was recorded in the donor, but pregnancy was confirmed in the recipient, though spontaneous abortion occurred after 10 weeks. This case will give useful information for further study of in vitro fertilisation, but also raises many ethical issues.     

Complete Suppression of the Gut Microbiome Prevents Acute Graft-Versus-Host Disease following Allogeneic Bone Marrow Transplantation

The hypothesis that elimination of facultative and strict anaerobic microorganisms from the gastro-intestinal tract by antimicrobial drugs in the period of time around allogeneic bone marrow transplantation (BMT) prevents acute graft-versus-host disease (GVHD), was examined in a cohort of 112 children grafted between 1989 and 2002 for hematological malignancies. All patients received T-cell replete marrow from human leukocyte antigens (HLA) matched sibling donors under identical transplantation conditions. To eliminate microorganisms from the gastro-intestinal tract, total gastro-intestinal decontamination (GID) was applied by high doses of non-absorbable antimicrobial drugs while the graft recipient was maintained in strict protective isolation. About half of the children (51%) proved to be successfully decontaminated, and about half (49%) unsuccessfully. One recipient got acute GVHD in the first group and 8 in the second group (p = 0.013). The degree of success of total GID was decisive for the occurrence of acute GVHD, irrespective of the presence of other risk factors such as higher age of recipient and/or donor, female donor for male recipient and carriership or reactivation of herpesviruses. Our results demonstrate that successful total GID of the graft recipient prevents moderate to severe acute GVHD. We suppose that substantial translocation of gastro-intestinal microorganisms or parts of these, functioning as microbial-associated molecular patterns (MAMP''s), triggering macrophages/dendritic cells via pattern recognizing receptors (PRR''s) is prohibited. As a consequence the initiation and progression of an inflammatory process leading to acute GVHD is inhibited.     

Usefulness of a first transferred free flap vascular pedicle for secondary microvascular reconstruction in the head and neck

The authors found that a previously transferred free flap vascular pedicle, distal to the first microvascular anastomosis, can be used as a recipient vessel for an additional free flap transfer. Free flap transfers were performed by using the standard procedure in patients with head and neck cancer. The mean age of the patients was 62 years. Five patients were men and three were women. A second free flap was transferred for secondary primary head and neck cancer in two cases, facial deformity in two cases, osteomyelitis of the skull in two cases, recurrent cancer in one case, and exposure of a mandibular reconstruction plate in one case. The interval between the two operations was from 4 months to 12 years (median, 21 months). All secondary free flaps were performed successfully. In two cases, the external jugular vein proximal to the previously anastomosed site was used for venous drainage. In another case, additional venous anastomosis was performed for flap congestion. It became clear that a previously transferred free flap vascular pedicle could be used as a recipient vessel for microvascular anastomosis. This is an excellent procedure for additional free flap transfers.