Objective To explore the influence of cirrhotic portal hypertension and its complications on liver transplantation. Methods The literatures of the recent years on influence of hepatic cirrhotic portal hypertension on liver transplantation were reviewed. Results Splenomegaly, hypersplenism, portal vein thrombosis, portosystemic shunt and collateral flow in cirrhotic patients will increase the difficulty of liver transplantation and lead to more postoperative complications. Appropriate handling of these conditions can achieve a higher success rate of liver transplantation. Conclusion Correct management of end-stage cirrhotic portal hypertension and its complications can expand the indications of liver transplantation and improve long-term survival rates.
Objective To investigate the pathological characteristics of hepatic energy metabolism changes following hepatic inflow occlusion and the tolerant limit to ischemia in cholestatic rats.Methods On the day 7 after rats biliary obstruction, the survival rate, hepatic mitochondrial respiratory function, content of ATP, and the ketone body ratio in arterial blood were investigated following the different duration of hepatic ischemia and reperfusion with portal blood bypass.Results The survival rate on postoperative day 10 was 100%, 100% and 40% subjected to 30, 60 and 90min of hepatic vascular occlusion. The hepatic energy metabolic function was decreased markedly following ischemia, and was increased markedly on 24 hours following reperfusion subjected to 30, 60min of hepatic vascular occlusion, but it had less increase with 90min of hepatic vascular occlusion.Conclusion The hepatic energy metabolic function injury is reversible in cholestatic rats, and the rats can tolerate hepatic inflow occlusion within 60min, but the hepatic energy metabolic function injury is irreversible after 90min of hepatic occlusion.
Objective To validate the mechanism of effect of hepatic artery ischemia on biliary fibrosis after liver transplantation and the prevention method. Methods Eighteen male dogs were established into the concise auto orthotopic liver transplantation models and assigned into three groups randomly: hepatic artery ischemia (HAI) group, TBB group (transferred the blood by a bridge duct ) and control group, each group contained 6 dogs. After opening portal vein, the samples were cut from liver in each group at the time of 6 h, 3 d and 14 d. The pathological modifications of intrahepatic bile ducts were observed and expression of transforming growth factor-β1 (TGF-β1) were detected in the three times. Expressions of Smad3 and phosphate-Smad3 as well as mRNA of α-smooth muscle actin (α-SMA) in intrahepatic bile ducts were detected 14 d after opening portal vein.Results Compared with control group, the collagen deposition and lumens stenosis in biliary vessel wall were more obviously in HAI group. In TBB group, the pathological modifications were slighter compared with HAI group. The positive cell index of TGF-β1 reached peak on day 3 after opening portal vein, then decreased in TBB group, and which in HAI group kept increase and was significantly higher than that in TBB group （Plt;0.05）. The expression level of phosphate-Smad3 and transcriptional level of α-SMA mRNA were 1.04±0.13 and 1.12±0.55 in TBB group on day 14 after opening portal vein, which were significantly higher than those in control group (0.59±0.09 and 0.46±0.18) and lower than those in HAI group (1.82±0.18 and 1.86±0.73), the diversities among three groups were significant (Plt;0.05). There was not significant difference of expression of Smads among three groups (Pgt;0.05). Conclusions Hepatic artery ischemia could increase the deposition of collagen fibers and the transdifferentiation of myofibroblast in bile duct and result in the biliary fibrosis by activating the TGF-β1/Smads signaling pathway. The bridging bypass device could lessen the biliary fibrosis caused by hepatic artery ischemia by inhibiting the activation of TGF-β1/Smads signal transduction passageway.
Objective To refine the technique of portal inflow occlusion and parenchymal transection for laparoscopic hepatectomy in the porcine model. Methods Ten pigs were used. The portal inflow complete or selective occlusion was carried out with portal triad clamping or dissection and division of the left portal pedicle. The sequential laparoscopic local hepatectomy, left lateral lobectomy, and left medial lobectomy were performed without portal inflow occlusion. Parenchymal transection was performed with harmonic scalpel, LigaSure, microwave dissector, bipolar electrocautery, surgical clips, and endoscopic stapler. The efficacy and safety of different techniques in laparoscopic parenchymal transection of the liver were compared. Results The ischemic liver was darken with complete or selective portal triad clamping. The ischemic demarcation line between left and right lobe was obvious with the dissection and division of the left portal pedicle. There was an applicable scope of each hepatic parenchymal transection apparatus. The optimal combination of different techniques could increase efficacy and reduce hemorrhage in laparoscopic parenchymal transection of the liver. Conclusion Technical refinements of portal inflow occlusion and parenchymal transection in porcine models could provide evidences to clinical appliance of laparoscopic anatomic major hepatectomy.
Objective To examine the effects of newly designed LY52 on the expression of matrix metalloproteinases and invasive ability of hepatocellular carcinoma HepG2 cells. Methods The effects of LY52 on the proliferations of HepG2 cells were detected by MTT assay. Gelatin zymography and Western blot were used to detect the effects of LY52 on matrix metalloproteinase-2 expression in the cell line. Transwell chamber assay was used to detect the effects of LY52 on the invasion of the cells. Results No obvious inhibitory or cytotoxicity effects of LY52 was found in lower concentrations (lt;200 μg/ml) of LY52. Gelatin zymography and Western blot showed that matrix metalloproteinase-2 expression were inhibited by LY52 in a dose-dependent manner in HepG2 cells. Furthermore, transwell chamber assay showed that LY52 could significantly inhibit the invasion of the cell line in a dose-dependent manner.Conclusion The results suggest that LY52 may inhibit the invasion of hepatocellular carcinoma cells by suppressing the matrix metalloproteinase-2 activity.
Objective To construct gene-modified hepatic stem cells (WB-F344 cells), which have rat IL-13 gene and can secrete the recombinant rat IL-13 cytokine in the cells. Methods Firstly, the rat IL-13 sequences were synthesized. Then the sequences were amplificated in bacterium coli after recombinated with pWPXL-MOD plasmid. After PCR and sequence identification, the positive clones were packaged into lentivirus. After detecting the virus titer, the WB-F344 cells with constructed lentivirus vector with rat IL-13 gene were cultured, then the valid targets (expression level of the IL-13) were detected by real time-PCR and Western blot in cultured WB-F344 cells on 5 days. Results The valid DNA of rat IL-13 was recombinated and packaged in lentivirus vector. The recombinant gene sequence was correct by checking with gene sequence test. Then the recombinant was introducted into the WB-F344 cells cultures. The best multiplicity of infection (MOI) value for effective transfection was 5. IL-13 had been detected on day 5 after transfection by checking with real-time PCR and Western blot. Conclusion The recombinant rat IL-13 gene with lentivirus vector is constructed and gene-modified WB-F344 cells are cultured successfully, which can be used in next animal experiment.
Objective To establish the model of hepatic VX2 tumor in rabbits and to offer the experimental evidences for the application of the model. Methods The hepatoma model was reproduced with VX2 cell lines in rabbits. The method to reproduce the model was improved. The changes of liver function (ALT, AST and TB) were determined at a different phase. Tumor’s growth and metastases, pathological changes, images and spontaneous survival time of the animal were observed. Results The tumors could grow up to 1.5-2.0 cm in diameter in 3 weeks after implanting. The successful rate of implantation was 100%. Nodular enhanced echo was found in the liver by color ultrasound. CT scans showed the low density foci in liver, while enhanced CT scans demonstrated asymmetrical intensification in the foci. Macroscopic observation showed that the tumors were grayish white in color and felt harder, necrotic foci was present in the center of tumor. Observation with light microscope showed that the tumor cells’ nucleoplasm proportion was great, tumor cells arranged irregularly, and the tumors displayed invasive growth and no obvious envelope around them. Animals’ spontaneous survival time was 40-53 days. The cause for their death was multiple system organ failure. Conclusion In pathological morphology, pathological process and prognosis, the hepaticVX2 tumors in rabbits are similar to human hepatocarcinoma. It has such characteristics as easy reproduction, short growth period, high success rate, high stability and so on. The model is an ideal hepatoma model in animals.
Objective Biliary epithelial cell (BEC) proliferated actively induced by ischemia-type biliary lesion (ITBL), which played an important role in the development of biliary complication after orthortopic liver transplantation (OLT). The aims of this study is to provide novel method to protect the liver endured cold preservation and reperfusion injury (CPRI) and reduce posttransplant biliary complication, and explore its possible mechanism.Methods Based on constructed OLT models for studying ITBL, the hepatic oval cell (HOC) or the IL-13 genemodified HOC to the portal vein of the recipient 〔OLT+HOC group and OLT+IL-13· HOC group〕 were-transfused, then the pathology change, the liver function and the expressions of the α-smooth muscle actin (αSMA) and Heme oxygenase-1 (HO-1) mRNA of the transplanted liver of CPRI were observed, the proliferation of BEC and survival rate of the recipients were also observed. Results The BEC injury was showed in grafts with prolonged ischemia time, characterized by induction of BEC proliferation, liver function injury and cholestasis sign reflecting the increase of serum ALT, AST and TBIL. The OLT+IL-13·HOC group had better results than OLT and OLT+HOC group, which indicated the OLT+IL-13·HOC group had low level of expression α-SMA (after operation 7 d, Plt;0.05) and proliferation of BEC (after operation 3 d, Plt;0.05). The expressions of HO-1 mRNA were higher in OLT+IL-13·HOC group than in other groups. The survival rate of OLT group was lower than that of the OLT+IL-13·HOC group and sham operation group (Plt;0.05).Conclusion High expression level of IL-13 in recipient rats could promote the expression of HO-1 mRNA in transplant liver, and profit to protection donor liver, and recover of the liver function after liver transplantation. It perhaps is the mechanism of protective effect of IL-13 on graft that stimulate the expression of HO-1 mRNA significantly.
【Abstract】Objective To establish animal model of orthotopic liver transplantation(OLT) in miniature pigs with high standardization, reproducibility and stability. Methods OLTs were performed without venovenous bypass in Bama miniature pigs. The survival rates and the changes of hemodynamics and metabolism were investigated. Results Twenty OLTs were performed between pairs of miniature pigs. The mean operative time and anhepatic phase were (181±25.8) min and (28.4±3.2) min respectively. During the anhepatic phase, dramatic hemodynamics and metabolism changes accompanied hyperkalemia identified. MAP and CVP decreased from (14.59±1.68) kPa and (0.66±0.11) kPa to (5.87±0.91) kPa and (0.27±0.10) kPa respectively, while temperature, pH, BE and HCO3- were significantly reduced (P＜0.05) and HR and K+ in serum were remarkable increased. After reperfusion, the disorder of hemodynamics and metabolism described above recovered gradually. 1week survival rate was 90%. Sixteen animals survived more than 2 weeks. AST, ALT and TBIL were significantly increased and reached the peak level on postoperative 1 day. From postoperative 2 day, AST, ALT and TBIL began to decrease and reached postanaesthesia level on postoperative 7 day. Conclusion The animal model of OLT without venovenous bypass in miniature pig, with its high standardization, reproducibility and stability, is an ideal one for series studies related to liver transplantation.
Objective To develop hepatic surgical planning software for hepatic operation on deciding the rational operational scheme and simulating procedures before the operation to accomplish the precise liver resection and decrease the operational risk. Methods3D-econstruction of liver was restored from spiral computed tomography (CT) data by using LiVirtue software. The liver and its anatomic structures were reconstructed to illuminate the location of the tumor and its related vessels to design a rational operational scheme. The virtul results, such as liver volume, hepatic sections, anatomy of portal vein and hepatic veins or possible operation plans, were compared with the actual situations during the operations. Results3D models of liver, tumor and their relative vessels were reconstructed successfully. Preoperative planning and intra-perative navigation based on the models ensured the safety of liver resection in our 32 cases of right lobe tumors. This preoperative simulation allowed surgeons to dissect the liver with reduced complications. These models could be also viewed and manipulated on personal computers.ConclusionThe LiVirtue is very helpful in the hepatic surgery, for clearly disclosing hepatic structures, rationally deciding operation schemes, virtually simulating the operations. This preoperative estimation from 3D model of liver benefits a lot to complicated liver resection.