[Experimental study on mechanism of graft injury of small-for-size liver transplantation in miniature pigs].

OBJECTIVE: To elucidate the mechanisms of graft injury in small-for-size liver transplantation. METHODS: Animal models were established with skeletonized and denervated anatomic parahepatic dissection, hepatectomy and perfusion in situ. Chinese Bama miniature pigs were divided into three groups (n = 5): Group A, liver transplantation; Group B, partial liver transplantation with right hemi-liver graft and Group C, liver transplantation with right median and caudate lobe graft. Animals were followed for 7 days with regards to survival, dynamical portal venous pressure (PVP), portal blood flow (PBF) and graft histopathological examination. RESULTS: Animal survivals were as follows: Group A, 5/5, Group B, 5/5 and Group C, 1/5. PVP rose immediately after reperfusion. PVP in Group C peaked to 28.6 +/- 2.07 mm Hg. Portal blood flow (PBF) measured by CDFI showed that the index of PBF per gram liver tissue reached 3.56 +/- 0.11 ml x min(-1) x g(-1) at the first hour post-reperfusion in Group C. Hepatic morphological examination showed that severe pathological changes occurred in small-for-size grafts, including sinusoidal congestion, hemorrhage, hepatocytic ballooning change or necrosis, endothelial cell detachment, Disse's space widening or vanishing and significant apoptosis. CONCLUSION: Portal over perfusion and acute portal hypertension are the primary etiological mechanisms of graft injury in small-for-size liver transplantation.

[Surgical treatment for hilar cholangiocarcinoma of Bismuth-Corlette type IV].

OBJECTIVE: To investigate the surgical treatment for hilar cholangiocarcinoma(HCCA) of Bismuth-Corlette type IV. METHODS: The clinical data of 73 HCCA patients admitted in Southwest Hospital, the Third Military Medical University from January 2002 to December 2008 were analyzed retrospectively. There were 41 males and 32 females, aged from 30 to 84 years old (averaged, 56.8 years old). All patients were diagnosed as hilar mass with hilar biliary obstruction by B-ultrasound, CT, magnetic resonance imaging, endoscopic retrograde cholangiopancreatography, magnetic resonance cholangiopancreatography or percutaneous transhepatic cholangiography, and confirmed by pathological examination in intra-operation and post-operation. Diagnosis of all patients were according to Bismuth-Corlette type IV. RESULTS: Fifteen cases underwent percutaneous transhepatic cholangial drainage, stents were implanted in 8 patients. Simple internal drainage was performed on 25 of the 73 patients and 4 with palliative resection. Radical resection was performed on 19 patients. The radical resection rate of HCCA were 26.0%. The 1, 3 years of survival rates were 36.8%, 10.5% respectively. The 1 year survival rate was 6.3% in drainage group. CONCLUSION: Radical resection is the potentially curative treatment for HCCA, a sufficient, reasonable use of internal and external drainage would improve the patient's quality of life.

[Protective effects of affiliating portasystemic shunt on graft injury in small-for-size liver transplantation in miniature pigs].

OBJECTIVES: To evaluate the protective effects of affiliating portasystemic shunt on small-for-size graft in liver transplantation. METHODS: Fifteen Chinese Bama miniature pigs were divided into three groups: group A (small-for-size liver transplantation), group B (distal splenorenal shunt + small-for-size liver transplantation), and group C (mesocaval H-shape shunt + small-for-size liver transplantation). Animals were followed up for 7 days with survival, dynamical liver function biochemical parameters, liver biopsies, portal venous pressure (PVP) and portal blood flow (PBF). RESULTS: Animal survivals were as follows: group A, 1/5, group B, 3/5 and group C, 5/5.Group A resulted in abnormal liver function parameters that were significantly ameliorated in group B and C. The histological examination of graft in group A displayed severe pathologic changes including hepatocyte vacuolar change or necrosis, sinusoidal congestion, parenchymal hemorrhage. Affiliating portasystemic shunt significantly alleviated graft injuries in group B and C. PVP rose and peaked up to 28.6 mm Hg (1 mm Hg = 0.133 kPa), PBF fluctuated after reperfusion in group A, but group B and C with affiliating portasystemic shunt showed significantly lower PVP and maintained rather stable PBF after reperfusion. There were also statistical differences in PVP or PBF between group B and C. CONCLUSIONS: Affiliating portasystemic shunt effectively might protect small-for-size graft from injuries after reperfusion.

Cold preservation of pig liver grafts with warm ischemia and pentoxifylline-UW solution.

BACKGROUND: We undertook this study to investigate the safe time limits of cold preservation in UW solution of liver grafts subjected to warm ischemia (WI) for 20 min and the changes of the limits when pentoxifylline is added to UW solution. METHODS: The safe time limit was studied in a simple porcine orthotopic liver transplantation (LTx) model. In donors, livers were subjected to 20 min of WI and subsequent 12-h (group 1, n = 5), 16-h (group 2, n = 5), and 20-h (group 3, n = 3) cold preservation in UW solution, respectively. After the safe time limits were clear, another group (group 4, n = 5) was built to test whether or not the limits can be changed when pentoxifylline is added to UW solution in an unsafe time limit group. RESULTS: All five animals in group 1 survived up to 7 days of the survey endpoint. In group 2, only one animal survived up to the same survey endpoint and all animals in group 3 died within 12 h. The 1-week survival rate of group 1 was significantly higher than the other two groups. Group 1 had a lower level of alanine aminotransferase (ALT) or aspartase aminotransferase (AST) after LTx, less pathological damage, higher concentration of adenosine triphosphate (ATP) and higher microcirculation blood flux in the grafted liver tissue at 1 h after reperfusion than the other two groups. The results primarily showed that 12-h cold preservation was safe, 16 h was unsafe, and 20 h was highly unsafe. But when pentoxifylline was added to UW solution in cold preservation (16-h group, group 4), in contrast to group 2, the incidence of liver tissue necrosis and primary graft nonfunction was significantly lower in group 4 than in group 2. The 1-week survival rate of the pigs was 100% in the former and 20% in latter group. Levels of ALT and AST in recipients' artery blood, malondialdehyde and TNF-alpha concentration in grafted liver tissue, resistance of portal vein and hepatic artery after preservation in group 4 were significantly reduced, whereas microcirculation blood flux of the grafted liver, superoxide dismutase concentration and ATP concentration in grafted liver tissue were significantly elevated. CONCLUSIONS: The safe time limit of cold preservation in UW solution of liver grafts subjected to WI for 20 min was about 12 h and the limits can be prolonged to 16 h when pentoxifylline is added to UW solution. Many mechanisms were involved.

Clinical analysis of surgical treatment of portal hypertension.

AIM: To review the experience in surgery for 508 patients with portal hypertension and to explore the selection of reasonable operation under different conditions. METHODS: The data of 508 patients with portal hypertension treated surgically in 1991-2001 in our centers were analyzed. Of the 508 patients, 256 were treated with portaazygous devascularization (PAD), 167 with portasystemic shunt (PSS), 62 with selective shunt (SS), 11 with combined portasystemic shunt and portaazygous devascularization (PSS+PAD), 9 with liver transplantation (LT), 3 with union operation for hepatic carcinoma and portal hypertension (HCC+PH). RESULTS: In the 167 patients treated with PSS, free portal pressure (FPP) was significantly higher in the patients with a longer diameter of the anastomotic stoma than in those with a shorter diameter before the operation (P < 0.01). After the operation, FPP in the former patients markedly decreased compared to the latter ones (P < 0.01). The incidence rate of hemorrhage in patients treated with PAD, PSS, SS, PSS+PAD, and HCC+PH was 21.09% (54/256), 13.77 (23/167), 11.29 (7/62), 36.36% (4/11), and 100% (3/3), respectively. The incidence rate of hepatic encephalopathy was 3.91% (10/256), 9.58% (16/167), 4.84% (3/62), 9.09% (1/11), and 100% (3/3), respectively while the operative mortality was 5.49% (15/256), 4.22% (7/167), 4.84% (3/62), 9.09% (1/11), and 66.67% (2/3) respectively. The operative mortality of liver transplantation was 22.22% (2/9). CONCLUSION: Five kinds of operation in surgical treatment of portal hypertension have their advantages and disadvantages. Therefore, the selection of operation should be based on the actual needs of the patients.

Etiology and surgical treatment of hilar bile duct stricture.

OBJECTIVE: To improve the surgical effects of hilar duct stricture. METHODS: The clinical data of 76 patients with hilar bile duct stricture treated at our hospital from 1990 to 2000 were analyzed. The diagnosis was determined by triad signs of cholangitis, increase of ALP and gamma-GGT levels, dilation of intrahepatic and extrahepatic bile ducts confirmed by ultrasonography (US), computed tomography (CT), percutaneous transhepatic cholangiography (PTC) or endoscopic retrograde cholangiopancreatography (ERCP). The location of stricture was divided according to the Bismuth classification standard. RESULTS: Among the 76 patients, 46 (60.5%) suffered from injurious stricture, including 13% of Bismuth type I, 39% of type II, 19.4% of type III, and 28.2% of type IV. Inflammatory stricture was found in 28 patients, locating in the left hepatic duct (LHD) 46.4% (13/28), the right hepatic duct (RHD) 35.7% (10/28), and the common hepatic duct (CHD) 17.9% (5/28), respectively. The percentages of patients with stricture due to Mirizzi's syndrome, bile duct cyst, and sclerosing cholangitis were 9.2%, 3.9% and 2.6%, respectively. Bile duct repair procedures included biliary reconstruction with pedicled umbilical vein graft for 9.2% of the patients, and proximal cholangiojejunostomy combined with LHD and RHD plasticity for 92.2%. Seventy of the 76 patients were followed up for 2-10 years, and the excellent outcome rate was 94.7%. CONCLUSIONS: Injurious stricture is the major type of hilar bile duct stricture. Inflammatory stricture is mainly composed of RHD. Hilar bile duct stricture should be treated surgically according to various etiological features and technical principles of biliary repair.

[Immune response of dendritic cells acquiring antigens from apoptotic cholangiocarcinoma cells induced by mitomycin].

OBJECTIVE: To investigate the anti-tumor immune response of dendritic cells (DCs) acquiring antigens from apoptotic cholangiocarcinoma cells and their therapeutic effects on cholangiocarcinoma cells. METHODS: DCs from human peripheral blood monocytes which acquired antigen capturing and processing capacity, characteristics of maturation, were established in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then cholangiocarcinoma cells were induced to apoptosis with mitomycin. The three groups included (1) coculture of DCs, apoptotic cancer cells and T cells, (2) coculture of DCs, necrotic cancer cells and T cells, (3) coculture of DCs, cultured cancer cells and T cells. After 7 days, DCs and T cells were riched separately to perform anti-tumor cells test and immune response test. RESULTS: these cells had typical dendritic cell morphology, expressed high levels of CD1a and B7, acquired antigen from apoptotic cells caused by mitomycin and could stimulate T cells to inhibit, even kill cholangiocarcinoma cells. CONCLUSIONS: The DCs from peripheral blood monocytes induced by GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by mitomycin, and stimulate T cells activity obviously. It maybe become an effective therapy for tumor.

[Screening and identification for cDNA of differentially expressed genes in human primary hepatocellular carcinoma].

OBJECTIVE: Screening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC). METHODS: The differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis. RESULTS: Thirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC. CONCLUSION: The subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC.

Effect of aging on cytoskeleton system of Kupffer cell and its phagocytic capacity.

AIM:To investigate the age-related alterations of cytoskeleton system in liver Kupffer cell and their relation to the changed phagocytic function.METHODS: The phagocytic function of Kupffer cells from rats of various ages (6mo, 12mo,18mo and 24mo) were quantitatively evaluated by phagocytosis of polystyrene beads. The actin distribution and measurement of Kupffer cell were determined by a phalloidin-TRITC method; and the myosin and vimentin distribution and measurement with indirect immunochemical staining.RESULTS: Aging resulted in significant alterations of actin, myosin and vimentin distributions and reductions in Kupffer cell; the 3 cytoskeleton components of 24-mo-old Kupffer cell were significantly decreased to 68.0%, 84.9% and 75.5%, respectively of these of 6-mo-old Kupffer cell(P < 0.01,0.01 and 0.01). And these decreases had significant positive relations with the damaged phagocytosis of the aged Kupffer cell.gammavalues were 0.96(P < 0.05), 0.99(P < 0.01) and 0.95 (P < 0.05) respectively.CONCLUSION: The cytos-keleton system of the aged Kupffer cell presents an evident state of senescence, which may be an important mechanism of decreased phagocytosis of the aged Kupffer cell.