大鼠肝小肠联合移植肠壁酶活性变化的研究

目的探讨肝小肠联合移植后肠壁酶活性的变化及其与移植肠功能和免疫排斥反应之间的关系。方法用封闭群大鼠建立肝小肠联合移植模型,术后定期对移植组织进行病理学、酶组织化学检查。结果单独小肠移植组术后均发生排斥反应,肠壁酶活性消失。肝小肠联合移植组５６％可避免排斥反应,肠壁酶活性和神经成分得以保持和恢复。结论肝小肠联合移植可使供肠避免或推迟被排斥。术后检测肠壁酶活性和神经成分的变化可用于监测排斥反应     

乙肝型炎肝硬化患者肝移植后近期肝肾脾功能的变化

目的:观察乙型肝炎肝硬化患者肝移植前后肝、肾及脾功能的变化。方法:行肝移植治疗的11例乙型肝炎肝硬化患者,移植前后检测肝、肾及脾功能,分析其变化。结果:凝血酶原时间:术后2d已显著低于术前,术后5d正常;球蛋白:术后1d正常,2d显著低于术前;肌酐:术后1d显著高于术前,7d恢复正常;血小板:术后3d显著下降,14d显著高于术前,21d正常。肾功能不全发生率72.3%,但均能恢复。结论:肝移植后,凝血酶原时间和球蛋白恢复正常,肌酐上升后短时间内可恢复正常,血小板下降后又恢复正常。     

Host immune suppression after small bowel/liver transplantation in rats

Simultaneous liver grafting in the Lewis (RT1¹)-to-DA (RT1^a) rat strain combination protects small intestinal grafts from rejection. The present study examined host immune responses after combined small bowel/liver transplantation (SBL) in this model. Orthotopic liver transplantation and heterotopic small intestinal transplantation were performed simultaneously and compared with isolated small bowel allografts (SBA) and isolated small bowel isografts (SBI). All rats were sacrificed on postoperative day (POD) 7 or 14 for immunological and histological studies. The mean time to rejection of the SBA was 6.6±0.3 days. Incontrast, there was no clinical or histological evidence of intestinal rejection in SBL recipients during the 14 days of follow-up. The SBL recipients showed clinical and histological evidence of graft-versushost disease (GVHD). Lmphocyte proliferation and IL-2 production in response to donor antigens were suppressed after SBL transplantation compared with the SBA or the SBI controls (P<0.05). Cell-mediated cytotoxicity and lymphocytotoxic antibody production against donor cells were also significantly inhibited in the SBL recipients compared with the SBA control group (P<0.05). We conclude that SBL transplantation in the Lewis-toDA rat strain combination: (1) suppresses host alloimmune responses, (2) prevents early intestinal rejection, and (3) favors the development of GVHD.     

蛙皮素对大鼠小肠移植黏膜免疫的影响

目的探讨蛙皮素对大鼠小肠移植肠道免疫的影响。方法本实验通过建立大鼠小肠移植模型,术后3d应用FK506。皮下微量注射蛙皮素后用流式细胞仪检测移植肠肠黏膜的淋巴细胞的T细胞的亚型B淋巴细胞的含量和组成的变化,观察蛙皮素对小肠移植术后移植肠肠免疫的影响。结果黏膜固有层BBS+FK506组CD3+、CD8+、CD45RA+细胞较生理盐水+FK506组明显增加(P<0.05)。Peyerpatches淋巴结BBS+FK506组CD3+、CD8+、CD45RA+细胞较生理盐水+FK506组明显增加(P<0.05)。CD4+细胞两组改变不明显。结论BBS可以明显改善小肠移植和免疫抑制剂造成的移植肠非特异性免疫细胞的减少,而不特异性免疫细胞的减少无明显改善,有助于小肠移植术后感染的防治。     

The early changes of colon motility in the rats after liver transplantation

The purpose of this study was to explore the early changes of colon motility in rats after liver transplantation. Thirty Wistar rats were divided into a sham operation group (n = 10) and a liver transplantation group (n = 10 pairs). The number of stools, the contractility of muscle strips, the length of smooth muscle cells, the levels of plasma endotoxin, the morphological changes, and the expression of inducible nitric oxide synthase (iNOS) in the colon of the rats were observed in the two groups. N(6)-(iminoethyl)-L-lysine (L-NIL, a selective iNOS antagonist) was used to confirm the activity of iNOS in the contractility of colonic motility. We observed the changes of nitrogen monoxide (NO) in plasma and colon mucosa of the two groups. Results showed that the liver transplantation group compared with the sham operation group showed significantly decreased contractility of the colon with significant differences in the morphological changes in rat colon. The expression of iNOS protein and iNOSmRNA was significantly increased in the liver transplantation group. The concentrations of plasma and colon mucosa NO and the levels of endotoxin were higher among the liver transplantation group than the sham operation group (P < .05). When the strips and cells of smooth muscle from the liver transplantation group were treated with L-NIL, their contractility increased. We concluded that colon motility decreased in the rats after transplantation, which could be related to the levels of plasma endotoxin and iNOS expressing in the colon.     

Early complications after orthotopic liver transplantation

The cost and impact of early post-transplant complications continue to be high. Diagnosis and management involves a high index of suspicion, rapid diagnostic and therapeutic interventions, and elimination of technical problems. Preoperative assessment of the donor and recipient medical condition and meticulous attention to detail during the technical performance of OLTx are the mainstays in achieving a good outcome.     

原位肝移植中移植肝早期功能的评价

目的通过测定原位肝移植中一些血清学指标来评价肝移植术后早期移植肝的功能状况。方法随机选取瑞金医院肝移植中心原位肝移植患者20例。分别在麻醉开始后（S）、无肝期开始即刻（A1）、无肝期结束前即刻（A2）、门静脉血流开放即刻（R0）、门静脉血流开放60min（R1）、门静脉血流开放3h（R3）、门静脉血流开放6h（R6）、门静脉血流开放12h（R12）、门静脉血流开放24h（R24）时间点抽取外周血，测定血清ALT、AST、LDH、GMP-140、vWF和HA。结果ALT、LDH在再灌注后即刻（R0）、AST在R6即达到峰值（P〈0．05），以后逐渐下降，但仍维持较高水平。LDH与ALT呈高度正相关（R=0．948，P〈0．001），LDH与AST呈中度正相关（R=0．646，P〈0．001）。GMP-140再灌注后开始升高，R3、R6、R12点达到峰值（P〈0．001），以后逐渐下降，但仍维持高水平。血管性血友病因子（von Willebrand factor，vWF）在再灌注后R6、R12、R24点高于正常。HA在再灌注后均高于正常。结论AST和ALT仍然是原位肝移植术后早期反映移植肝功能状况的可靠指标；LDH与ALT及AST存在良好相关性；GMP-140、vWF和HA的升高反映了肝窦内皮细胞（SEC）的损伤程度，但能否准确反映早期移植肝功能状况则有待进一步证实。     

肝硬化鼠肝移植后早期全身和内脏血流动力学的变化

目的 观察正常鼠肝移植及及肝硬化鼠肝移植术早期全身和内脏血流动力学的变化。方法 实验动物随机分为正常鼠（NL,10只）、肝硬化鼠（IHPH,10只）、正常鼠肝移植（NL－OLT,9只）、肝硬化鼠肝移植（IHPH－OLT,16只）组。分别采用放射性微球法行血流动力学研究。结果 NLOLT鼠绝大多数血流动力学参数与NL鼠比较差异无显著意义。IHPH及IHPH－OLT 3d,7d组心输量和内脏血流量和内脏血流量增加,平均动脉压、周围血管总阻力和内脏血管阻力降低。内脏血流动力学紊乱较全身明显。结论 肝硬化鼠肝移植后的血流动力学紊乱可能与移植前已存在的病理生理因素有关。     

Surgical technique for combined liver/intestine transplantation in rats.

Simultaneous liver transplantation may reduce the risk of intestinal transplant rejection. We have recently developed two new models of combined liver/intestine transplantation (LIT) in the rat to study this phenomenon. Herein, we report our experience with LIT using a single donor (SD) or multiple donors (MD). Large volumes of fluid were required to prevent a drop in the mean arterial pressure during the anhepatic phase in the SD recipients. Many of the SD recipients died of intraoperative hypovolemic shock (57%). The MD recipients had a shorter anhepatic time (12 +/- 1 minutes vs. 17 +/- 2 minutes; P less than 0.01) and a shorter warm intestinal ischemia time (15 +/- 1 minutes vs. 32 +/- 2 minutes; P less than 0.01). Operative mortality rates were much lower in the MD recipients (10% vs. 68%; P less than 0.01). The long-term survival rate using MD was 71% at 1 month. Graft function was normal in the long-term survivors. LIT with MD provides a good model to study the immunological effects of multivisceral grafting.     

Early vascular complications after pediatric liver transplantation

Vascular complications have a detrimental effect on the outcome after liver transplantation. Most studies focus exclusively on hepatic artery thrombosis (HAT). The current study analyzed the incidence, consequences, and risk factors for HAT, portal vein thrombosis (PVT), and venous outflow tract obstruction (VOTO) in a consecutive series of 157 pediatric liver transplantations. The overall incidence of vascular complications was 21%. The incidences of HAT, PVT, and VOTO were 10%, 4%, and 6%, respectively. Patient survival after PVT and VOTO and graft survival after HAT and PVT were less compared with survival of grafts without vascular complications. To identify risk factors for vascular complications, factors related to recipient, donor, and surgical techniques were analyzed. A low donor-recipient (D/R) age ratio, long surgical time, and use of the proper hepatic artery of the recipient for arterial reconstruction were risk factors for HAT Young age, low weight, segmental grafts, and piggyback technique were risk factors for PVT. Fulminant hepatic failure, high D/R age and weight ratios, and use of segmental grafts were related to VOTO. Vascular complications, which occurred in 21% of the pediatric liver transplantations, had a significant impact on patient and graft survival. Size disparity between donor and recipient was an important risk factor for vascular complications, especially in the case of transplantation of segmental grafts. Patient and graft survival might improve by avoiding the identified risk factors.     

Nitric oxide-related neural components in the rat small intestine after transplantation

Abstract The changes in nitric oxide (NO)-related neural components in the transplanted small intestine are unknown. In this study, the NO neural component was examined using electrophysiological and NADPH-diaphorase his-tochemical techniques in a rat small bowel transplantation model. Syngeneic total small bowel transplantation was performed in 26 male Lewis rats using microsurgical techniques. The rats were divided into four groups: an untreated control group and animals at 1 (G1), 2 (G2), and 4 (G4) weeks after transplantation. Jejunal strips were mounted in a superfusion apparatus for examination. In the presence of atropine and guanethidine, the effect of the NO synthesis inhibitor L-N^G-nitro-arginine (L-NNA, 100 µM) relaxation mediated by the nonadrenergic, noncholinergic (NANC) neural system was assessed following electrical stimulation at 2 Hz. The inhibitory effect of L-NNA on relaxation was taken as an indicator of NO production. The percentage of inhibition in the control group, and in G1, G2, and G4 was 43.30 %+ 6.08 % (mean ± SE), 42.10 %± 6.69 %, 43.62 ± 10.00 %, and 52.46 %+ 6.00 %, respectively. Inhibition in G4 was significantly higher than in the other groups ( P< 0.01). The percentage of NADPH-diaphorase-positive fibers in the control group, G1, G2, and G4 was 25.06%+4.70% (mean ± SE), 26.27 %± 2.17 %, 24.73%±2.87%, and 30.76 %± 3.19 %, respectively. Again, G4 showed a significantly higher level than the other groups ( P < 0.01). We conclude that increased NO production may play a significant role in maintaining the intrinsic nervous system of the small intestine after transplantation.     

Nitric oxide-related neural components in the rat small intestine after transplantation

The changes in nitric oxide (NO)-related neural components in the transplanted small intestine are unknown. In this study, the NO neural component was examined using electrophysiological and NADPH-diaphorase histochemical techniques in a rat small bowel transplantation model. Syngeneic total small bowel transplantation was performed in 26 male Lewis rats using microsurgical techniques. The rats were divided into four groups: an untreated control group and animals at 1 (G1), 2 (G2), and 4 (G4) weeks after transplantation. Jejunal strips were mounted in a superfusion apparatus for examination. In the presence of atropine and guanethidine, the effect of the NO synthesis inhibitor L-N^G-vitro-arginine (L-NNA, 100 M) relaxation mediated by the nonadrenergic, noncholinergic (NANC) neural system was assessed following electrical stimulation at 2 Hz. The inhibitory effect of L-NNA on relaxation was taken as an indicator of NO production. The percentage of inhibition in the control group, and in G1, G2, and G4 was 43.30% ± 6.08% (mean ± SE), 42.10%± 6.69%, 43.62±10.00%, and 52.46% ± 6.00%, respectively. Inhibition in G4 was significantly higher than in the other groups (P< 0.01). The percentage of NADPH-diaphorase-positive fibers in the control group, G1, G2, and G4 was 25.06 % ± 4.70% (mean ± SE), 26.27% ± 2.17% , 24.73% ± 2.87%, and 30.76% ± 3.19%, respectively. Again, G4 showed a significantly higher level than the other groups (P < 0.01). We conclude that increased NO production may play a significant role in maintaining the intrinsic nervous system of the small intestine after transplantation.     

Morphological and physiological changes of interstitial cells of Cajal after small bowel transplantation in rats

Intestinal dysmotility has been reported to be associated with a decreased number of interstitial cells of Cajal (ICCs). However, the chronological changes in ICCs after small bowel transplantation (SBT) have not yet been elucidated. In this study, we aimed to evaluate the chronological change of ICCs after SBT. Orthotopic syngeneic SBT was performed in rats. Graft specimens were obtained at postreperfusion, and on 1, 3, 7, 14, and 30 postoperative day (POD). Thereafter, immunohistochemical staining was performed and the spontaneous contractions measured. During the initial period after SBT, the temporal impairment of ICCs was found. In an immunohistochemical study, c-Kit-positive cells appeared to decrease on POD 0, 1, and 3. Thereafter, the number of cells increased gradually up to POD 7. In contrast, the recovery of the spontaneous contractile amplitude took more time. The frequency of the electrical signal was preserved at almost exactly the same levels throughout this experimental period. Although the network of ICCs was found to be temporarily impaired after SBT in an immunohistochemical examination, this change was reversible. Moreover, the recovery of the function of the intestinal motility associated with ICCs was delayed after the early postoperative period.     

Experimental small bowel transplantation using newborn intestine in rats: I. Lipid absorption restored after transplantation of nonvascularized graft.

BACKGROUND/PURPOSE: Utilizing the characters of neovascularized activity of newborn organs, the authors developed a rat model of small bowel transplantation with a free graft of newborn intestine into the recipient's omentum. METHODS: Segmental intestine from newborn rats were grafted into the omentum without vascular anastomosis in a syngeneic combination (n = 19). The transplanted intestine was examined morphologically and electrophysiologically 4 weeks after grafting. Then, recipients' small intestine was totally substituted by the transplanted newborn intestine, and recipients' survival was recorded after orthotopical reconstruction. During the experimental periods, feces of these rats were collected, and total lipid excretion was measured. The short-gut rats, whose small bowel was totally resected, served as a control (n = 12). RESULTS: Thirteen of 19 grafts (68.4%) were judged as a histologically mature intestine. They showed typical slow waves that were identical to those of native small intestine. After all of the mature grafts were interposed, six recipients (46.2%) survived longer than 15 weeks. Control short gut animals severely lost weight and died except for one. CONCLUSION: Newborn intestinal transplantation could restore severe weight loss in the short-gut rats and save them.