Small bowel transplantation in mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.     

A Case of Budd-Chiari Syndrome Associated with Alveolar Echinococcosis

Although alveolar echinococcosis (AE) can cause a serious disease with high mortality and morbidity similar to malign neoplasms. A 62-year-old woman admitted to a hospital located in Sivas, Turkey, with the complaints of fatigue and right upper abdominal pain. On contrast abdominal CT, a 54×70×45 mm sized cystic lesion was detected in the left lobe of the liver that was seen to extend to the posterior mediastinum and invade the diaphragm, esophagus, and pericardium. The cystic lesion was seen to be occluding the inferior vena cava and left hepatic vein at the level where the hepatic veins poured into the inferior vena cava. Bilateral pleural effusion was also detected. We discussed this secondary Budd-Chiari Syndrome (BCS) case, resulting from the AE occlusion of the left hepatic vein and inferior vena cava, in light of the information in literature.     

Establishment of animal model of dual liver transplantation in rat

The animal model of the whole-size and reduced-size liver transplantation in both rat and mouse has been successfully established. Because of the difficulties and complexities in microsurgical technology, the animal model of dual liver transplantation was still not established for twelve years since the first human dual liver transplantation has been made a success. There is an essential need to establish this animal model to lay a basic foundation for clinical practice. To study the physiological and histopathological changes of dual liver transplantation, "Y" type vein from the cross part between vena cava and two iliac of donor and "Y' type prosthesis were employed to recanalize portal vein and the bile duct between dual liver grafts and recipient. The dual right upper lobes about 45-50% of the recipient liver volume were taken as donor, one was orthotopically implanted at its original position, the other was rotated 180° sagitally and heterotopically positioned in the left upper quadrant. Microcirculation parameters, liver function, immunohistochemistry and survival were analyzed to evaluate the function of dual liver grafts. No significant difference in the hepatic microcirculatory flow was found between two grafts in the first 90 minutes after reperfusion. Light and electronic microscope showed the liver architecture was maintained without obvious features of cellular destruction and the continuity of the endothelium was preserved. Only 3 heterotopically positioned graft appeared patchy desquamation of endothelial cell, mitochondrial swelling and hepatocytes cytoplasmic vacuolization. Immunohistochemistry revealed there is no difference in hepatocyte activity and the ability of endothelia to contract and relax after reperfusion between dual grafts. Dual grafts made a rapid amelioration of liver function after reperfusion. 7 rats survived more than 7 days with survival rate of 58.3.%. Using "Y" type vein and bile duct prosthesis, we successfully established a novel rat model of dual right upper liver lobe transplantation.     

经皮微波固化对肝内大血管旁复发性肝癌的疗效及安全性

目的:研究经皮微波固化治疗肝内大血管旁复发性肝癌的疗效和安全性。方法:选择2011年2月~2015年12月在我院进行诊治的不能进行手术切除的肝内大血管(肝静脉、肝后下腔静脉、肝动脉或门静脉)旁复发性肝癌患者80例,随机分为观察组和对照组,观察组进行经皮微波固化治疗,对照组进行手术再切除,于治疗前后检查甲胎蛋白、B超,观察术后并发症的发生情况。结果:观察组治疗后甲胎蛋白转阴16例,降低21例,总有效率为92.5%,明显高于对照组的85.9%(P0.05);治疗后3个月经超声检测肿瘤大小,观察组有15例肿瘤停止生长,21例肿瘤缩小,无效的4例中肿瘤合并肝内转移或出现范围扩大,对照组有14例肿瘤停止生长,19例肿瘤缩小,7例无效,观察组控制肿瘤大小有效率明显大于对照组(P0.05);观察组并发症发生率为22.5%(9/40),明显低于对照组的35.0%(14/40)(P0.05)。结论:经皮微波固化治疗肝静脉、肝后下腔静脉、肝动脉或门静脉旁复发性肝癌疗效确切,术后并发症少,可用于临床肝内大血管旁复发性肝癌患者的治疗。     

Successful use of recombinant factor Viia in traumatic liver injury--a case report

The paper describe the use of rFVIIa in the management of massive bleeding in a patient with polytrauma involving liver injury. An 18-year-old girl with severe polytrauma sustained during a bus-car collision. She had multiple musculoskeletal injuries, severe concussion of the liver with amputation of the left liver lobe, disruption of the left hepatic vein from the inferior vena cava, and impaired hemostasis. Acute bleeding (> 5 L) was not improved by conservative methods and a single dose of rFVlla 90 pg/kg was administered. Infusion of rFVlla resulted in an immediate clinical effect with rapid improvements in blood laboratory measurements and coagulation parameters. rFVIIa should be considered as an adjunctive treatment for the control of hemorrhage in severely injured patients with uncontrolled bleeding and impaired hemostasis.     

Gluconeogenesis by the fetal sheep liver in vivo

Hepatic gluconeogenesis is an important source of glucose postnatally. Whether hepatic gluconeogenesis contributes to fetal glucose supply has not been studied directly in vivo. Previous studies of gluconeogenesis in fetal sheep have assessed total fetal glucose production, and the results have been controversial. To assess the specific role of the liver in gluconeogenesis in fetal sheep, we placed catheters in the right or left hepatic vein, umbilical vein and the inferior vena cava of six fetal sheep (mean gestational age 134 days) and infused a radioactive gluconeogenic substrate (14C-lactate or 14C-alanine) into the fetal inferior vena cava. We measured 14C-glucose radioactivity (dpm/ml) in the right or left hepatic vein and calculated the arteriovenous difference in 14C-glucose radioactivity (dpm/ml) across the right or left liver lobe. We found that only 0.35% of the 14C substrates perfusing either the right or the left hepatic lobe of the fetal liver were converted to 14C-glucose. Even when considerable glucose was released by the liver, the percentage of substrates converted to glucose remained very low (maximum 1.7%), indicating that gluconeogenesis did not contribute significantly to the glucose released. We conclude that gluconeogenesis by the fetal liver contributes negligibly to the glucose supply in fetal sheep.     

Dependence of the topography of the hepatic veins on the external shape of the liver, its size and on age

Hepatic preparations obtained from 186 newborns, children and mature persons have been studied. In newborns and babies the size of the right and left hepatic lobes are nearly the same. Beginning from an early age, the right hepatic lobe begins to grow, and the oblongatal or oval form of the liver changes into a triangle one. With changes in its size and form, the topography of the hepatic veins changes, too. In children younger than 1 year of age having the oblongatal or oval form of their liver, the diameters of the right, middle and left hepatic veins are nearly equal. In mature persons having the triangle form of the liver, the right hepatic vein is the largest and its diameter is twice as large as that of the left hepatic vein. Additional hepatic veins flowing into the inferior vena cava, are mainly formed by the branches participating in drainage of the right hepatic lobe. In 25 cases there are additional large hepatic veins; they are observed in cases when the right hepatic vein is poorly developed.     

Orthotopic Kidney Transplantation in Mice: Technique Using Cuff for Renal Vein Anastomosis

Mouse renal transplantation is a technically challenging procedure. Although the first kidney transplants in mice were performed over 34 years ago and refined some years later, the classical techniques of mouse renal transplantation required clamping both vena cava and aorta simultaneously and carry out suture anastomoses of the renal artery and vein in a heterotopic position. In our laboratory, we have successfully developed mouse orthotopic kidney transplantation for the first time, using a rapid “cuffed” renal vein technique for vessel anastomosis, wherein the donor’s renal vein was inserted through an intravenous catheter, folded back and tied. During grafting, the cuffed renal vein was directly inserted into the recipient’s renal vein without the need for the clamping vena cava and suturing of renal vein. This technique allowed for the exact transplantation of the kidney into the original position, compared to the classical technique, and has significantly shortened the clamping time due to a quicker and precise anastomosis of renal vein as described. This also allowed for a quicker recovery of the lower extremity activity, reduction in myoglobinuria with resultant kidney graft survival of 88.9%. Thus we believe that the cuffed renal vein technique simplifies microvascular anastomoses and affords important additional benefits.     

Portocaval shunt for hepatocyte package

In developing therapeutic alternatives to liver transplantation, we have used the strategy of applying a small intestinal segment as a scaffold for hepatocyte transplantation and also as a portocaval shunt (PCS) system to address both liver dysfunction and portal hypertension. The aim of this study was to investigate the feasibility of such an intestinal segment in animal models. Hepatocytes isolated from luciferase-transgenic Lewis rats were transplanted into jejunal segments of wild-type Lewis rats with mucosa removal without PCS application. Luciferase-derived luminescence from transplanted hepatocytes was stably detected for 30 days. Then, we performed autologous hepatocyte transplantation into the submucosal layer of an isolated and vascularized small intestinal segment in pigs. Transplanted hepatocytes were isolated from the resected left-lateral lobe of the liver. On day 7, hepatocyte clusters and bile duct-like structures were observed histologically. To create an intestinal PCS system in pigs, an auto-graft of the segmental ileum and interposing vessel graft were anastomosed to the portal vein trunk and inferior vena cava. However, thrombi were observed in vessels of the intestinal PCSs. We measured the correlation between infusion pressure and flow volume in whole intestines ex vivo in both species and found that the high pressure corresponding to portal hypertension was still insufficient to maintain the patency of the intestinal grafts. In conclusion, we demonstrated the feasibility of the small intestine as a scaffold for hepatocyte transplantation in rat and pig models, but PCS using an intestinal graft failed to maintain patency in a pig model.     

80％门静脉结扎后肝脏组织中MMP-9的表达与肝再生作用的相关性研究

目的：探讨基质金属蛋白酶-9（MMP-9）在大鼠80％门静脉分支结扎模型中大鼠增生肝脏组织中的表达及其与肝再生作用的关系。方法：健康SD雄性大鼠48只,随机平均分成假手术对照组（Sham）和门静脉结扎实验组（PVL）。观察术后1、3、7和14d保留侧肝叶重量／体重比值;下腔静脉采血后检测血清谷丙转氨酶（ALT）、谷草转氨酶（AST）值的变化;光镜下观察保留侧肝脏组织的病理形态变化;用免疫组化法检测增殖细胞核抗原（Proliferating cell nuclear antigen,PCNA）的表达,用免疫印记法检测MMP-9的表达,并进行统计分析。结果：80％门静脉分支结扎后,结扎侧肝叶呈进行性萎缩,保留侧肝叶重量／体重比值逐渐增加,7d达“平台期”;与对照纽明显不同,PVL组的ALT、AST的值在1h达到高峰,7d后回到正常水平;保留侧肝脏组织中PCNA阳性细胞计数与对照组比较,3d开始表述增强（P〈0．05）,7d以后逐渐恢复至正常水平（P〉0．05）;保留侧肝叶MMP-9蛋白的表达在术后3d开始增加,术后7d达到高峰。结论：MMP-9蛋白的表达在80％门静脉结扎后大鼠肝再生过程中发挥重要作用。     

Chronic utero-ovarian vein catheterization with subsequent occlusion prolongs the estrous cycle and changes electromyographic activity in the myometrium of ewes

The objective of our study was to determine the effect of chronic utero-ovarian vein catheterization in ewes on estrous cycle length, plasma progesterone (P) concentration, and myometrial electromyographic activity. Cyclic ewes with inferior vena cava catheters were used as controls. Estrus was synchronized in ten ewes and 10 to 12 d following estrus, the ewes were anesthetized, fitted with myometrial electromyograph leads and with utero-ovarian vein (n = 5) or inferior vena cava (n = 5) catheters. After surgery, ewes returned to estrus as expected (16 to 18 d interestrus interval). The second cycle of four of five ewes with utero-ovarian vein catheters were prolonged (40 to 58 d). The inferior vena cava catheterized ewes had normal length second cycles. Plasma P concentrations reflected the estrous cycles: low ( 0.05).     

80%门静脉结扎后肝脏组织中MMP-9 的表达与肝再生作用 的相关性研究

目的:探讨基质金属蛋白酶-9(MMP-9)在大鼠80%门静脉分支结扎模型中大鼠增生肝脏组织中的表达及其与肝再生作用的关系。方法:健康SD雄性大鼠48只,随机平均分成假手术对照组(Sham)和门静脉结扎实验组(PVL)。观察术后1、3、7和14d保留侧肝叶重量/体重比值;下腔静脉采血后检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)值的变化;光镜下观察保留侧肝脏组织的病理形态变化;用免疫组化法检测增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)的表达,用免疫印记法检测MMP-9的表达,并进行统计分析。结果:80%门静脉分支结扎后,结扎侧肝叶呈进行性萎缩,保留侧肝叶重量/体重比值逐渐增加,7d达"平台期";与对照组明显不同,PVL组的ALT、AST的值在1h达到高峰,7d后回到正常水平;保留侧肝脏组织中PCNA阳性细胞计数与对照组比较,3d开始表达增强(P<0.05),7d以后逐渐恢复至正常水平(P>0.05);保留侧肝叶MMP-9蛋白的表达在术后3d开始增加,术后7d达到高峰。结论:MMP-9蛋白的表达在80%门静脉结扎后大鼠肝再生过程中发挥重要作用。     

THE SURGICAL TREATMENT OF PORTAL HYPERTENSION

Portacaval shunt operations were done in 15 of 18 patients who were treated surgically for portal hypertension. In eight cases the shunt was established by splenorenal anastomosis; in seven by anastomosis of the portal vein to the side of the inferior vena cava. Of the eight patients with splenorenal shunt, two are well, four are considered improved, and two have died. Of the seven in whom the portal vein was joined to the vena cava, two are improved, one is unimproved, and four have died.     

Efficacy and Safety of Right Hemihepatectomy Through the Right Retrohepatic Tunnel

We wished to study the efficacy and safety of the retrograde ligation of short hepatic veins (SHVs) and the right hepatic vein (HV) through the retrohepatic tunnel in patients who underwent hemihepatectomy due to large hepatic carcinoma in the right lobe of the liver. Right hemihepatectomy was performed in 23 patients with tumors larger than 8 cm in diameter. The liver was separated at the secondary porta, and the interspace between right HVs and middle HVs was expanded. The right hepatic portal vein and hepatic artery were freed and ligated, followed by the retrograde dissection of SHVs and the right HV along the right retrohepatic space anterior to the inferior vena cava. A blocking belt was set at the left side of the midline, after which the right liver was cut off. The procedure was successfully completed in all patients. The average amount of intraoperative blood loss was 640 ml. The change in hepatic function was observed on the third postoperative day. Twenty-two patients exhibited satisfactory results; one patient died from postoperative hepatic failure. In conclusion, this procedure can be safely performed in most hemihepatectomy patients with liver tumors.     

Precise Reconstruction of Veins and Bile Ducts in Rat Liver Transplantation

Rat orthotopic liver transplantation (ROLT) remains a technically demanding procedure, especially regarding the reconstruction of the suprahepatic vena cava (SHVC). In this study, a new microsuture technique was developed for anastomosis of the SHVC, and a special single-groove cuff and blade-cut stent were introduced. With these modified techniques, we aimed to make a precise anastomosis of the SHVC and to provide optimal cuffs and stents for the reconstruction of the veins and bile ducts. According to different microsuture techniques for the SHVC and different types of cuffs and stents, three ROLT groups were created to compare the operation times and prognoses. Sham operations were performed as controls in the fourth group. The time expenditures with each step were compared among the transplantation groups. Biochemical parameters were tested at the end of a 1-month observation period. The short- and long-term survival rates of the transplantation groups were recorded and compared. Our new microsuture technique was faster than the conventional continuous suture technique for SHVC anastomosis (P < 0.05). The use of a single-groove cuff for reconstruction of the portal vein and the infrahepatic vena cava shortened the anastomotic time (P < 0.05). The use of blade-cut stents resulted in fewer biliary complications and better survival over the short and long terms (P < 0.05). Our new microsuture technique and the single-groove cuffs proved to be a precise method for venous reconstruction which shortened the anhepatic time and the anastomotic time significantly. The blade-cut stents apparently reduced the incidence of biliary complications. In summary, with this precise microsuture technique and delicate cuffs and stents, excellent long-term survival can be achieved easily and stably for ROLT.     

肝脏移植术前受体64 排CT 血管成像临床应用价值

目的:探讨肝脏移植术前受体64排螺旋CT血管成像(computed tomography angiography CTA)在临床的应用价值。方法:对28例晚期肝病患者进行64排增强后动脉期、门脉期及静脉期扫描,使用多种重建方法,由两名以上有经验的医师对肝脏血管进行分析评价,并与18例移植术后结果比较,探讨64排CT血管成像对肝移植术前准备的意义。结果:发现肝动脉变异6例,腹腔干起始部狭窄2例,腹腔干动脉瘤1例,脾动脉瘤2例,肝脏主要供血动脉直径<3mm 3例,肝门静脉海绵样变3例,肝外门静脉主干栓塞5例,伴有肝内门静脉栓塞3例,肝外门静脉主干栓塞伴肠系膜上静脉广泛栓塞2例,下腔静脉癌栓3例,其中癌栓达右心房2例。结论:64排CT血管成像无创、可靠,通过多种重建方法,能清晰显示肝脏血管,对肝脏移植术前血管做出评价,具有可靠的临床指导意义。     

Contribution of the umbilical and portal veins to the hepatic blood supply of guinea pig fetuses--an angiographic study.

The interlobular distribution of the umbilical and portal venous blood flow within the liver was examined in 35 guinea pig fetuses between 59 and 65 days of gestation. Contrast medium was injected into the umbilical or vitelline vein, and its passage through the liver was monitored by serial angiography. In four experiments, injections were made into both the umbilical and vitelline veins of the same fetus. To ease interpretation of the angiograms obtained in vivo, we also made a postmortem examination of livers in which the venous system had been filled with an aqueous suspension of barium sulphate in gelatin. These combined experiments demonstrated no passage of contrast medium from the placenta to the inferior vena cava, which is in accordance with independent evidence that the term guinea pig fetus lacks a functional ductus venosus. The area supplied by the umbilical and portal veins was clearly and consistently delineated. The umbilical vein supplied the left lobe and the left sublobe of the quadrate lobe. The portal vein supplied the right lobe, the smaller caudate lobe, and all or most of the right sublobe of the quadrate lobe. This pattern of distribution appears to be determined by flow and pressure gradients within the hepatic circulation.     

Simultaneous sampling of portal, hepatic and systemic blood during intragastric loading and tracer infusion in conscious pigs

A surgical model for catheterization at multiple sites has been developed for use in long-term metabolic studies. For blood sampling, catheters were inserted into the portal and hepatic veins and the common carotid artery. The hepatic vein catheter was inserted from the margin of a liver lobe and led through the venous system, until the tip was close to the bifurcation with the inferior vena cava. A new technique was developed to ensure correct placement of the hepatic vein catheter using the specific extraction of indocyanin-green over the liver during surgery. Gastrostomy was performed using a Pezzer catheter. Catheters in the artery and hepatic and portal veins were patent for blood withdrawal for up to 4 weeks, and thus allowed repeated metabolic studies. Studies were performed in conscious animals familiar with the experimental situation.     

Velocity profiles in the human ductus venosus: a numerical fluid structure interaction study

The veins distributing oxygenated blood from the placenta to the fetal body have been given much attention in clinical Doppler velocimetry studies, in particular the ductus venosus. The ductus venosus is embedded in the left liver lobe and connects the intra-abdominal portion of the umbilical vein (IUV) directly to the inferior vena cava, such that oxygenated blood can bypass the liver and flow directly to the fetal heart. In the current work, we have developed a mathematical model to assist the clinical assessment of volumetric flow rate at the inlet of the ductus venosus. With a robust estimate of the velocity profile shape coefficient (VC), the volumetric flow rate may be estimated as the product of the time-averaged cross-sectional area, the time-averaged cross-sectional maximum velocity and the VC. The time average quantities may be obtained from Doppler ultrasound measurements, whereas the VC may be estimated from numerical simulations. The mathematical model employs a 3D fluid structure interaction model of the bifurcation formed by the IUV, the ductus venosus and the left portal vein. Furthermore, the amniotic portion of the umbilical vein, the right liver lobe and the inferior vena cava were incorporated as lumped model boundary conditions for the fluid structure interaction model. A hyperelastic material is used to model the structural response of the vessel walls, based on recently available experimental data for the human IUV and ductus venous. A parametric study was constructed to investigate the VC at the ductus venosus inlet, based on a reference case for a human fetus at 36 weeks of gestation. The VC was found to be \(0.687\,\pm \,0.023\) (Mean \(\pm \) SD of parametric case study), which confirms previous studies in the literature on the VC at the ductus venosus inlet. Additionally, CFD simulations with rigid walls were performed on a subsection of the parametric case study, and only minor changes in the predicted VCs were observed compared to the FSI cases. In conclusion, the presented mathematical model is a promising tool for the assessment of ductus venosus Doppler velocimetry.     

Thromboses néoplasiques et thromboses cruoriques mécaniques compliquant un cancer du testicule gauche en TEP/TDM au 18FDG

We report the case of a 46-year-old patient with pulmonary embolism complicating thrombosis of the left renal vein and inferior vena cava, which came from a malignant tumor of the left testicle. The case has been elucidated by positron emission tomography with ¹⁸FDG coupled to the CT-scan (PET/CT), performed as part of the etiological assessment of this pulmonary embolism, and anatomical and physiological considerations. The examination revealed ¹⁸FDG uptake in thrombus of the left kidney vein and the inferior vena cava. In view of the anatomy of the left spermatic vein, this finding led to the left testicle, which shows no nuclide uptake but was bathed in a large hydrocele visible on CT images. If several cases of pulmonary embolism or thrombosis of the inferior vena cava revealing a testicular cancer have been described in the literature, there has been no report including ¹⁸FDG PET/CT to the best of our knowledge. This case illustrates the imaging differences between bland and malignant thrombus as a consequence of their pathophysiology. It highlights the contribution of CT images from the PET/CT for the diagnosis.