Objective　To study the mRNA expression of MCT1 and MCT2 genes in human hepatocellular carcinoma (HCC) and paracarcinoma liver tissue (PCLT). Methods　The semi-quantitative analysis of MCT1 and MCT2 genes mRNA expression in human HCC and PCLT was conducted by RT-PCR method and electrophoresis band opacity density (OD) comparison analysis method in 25 patients with HCC. Results　The mRNA expression of MCT1 was significantly higher than MCT2 in HCC and PCLT, in HCC the mRNA expression of MCT1 and MCT2 genes were significant higher than that in PCLT. Conclusions　The high expression of mRNA of MCT1 and MCT2 genes in HCC indicates that these genes may take a significant role on lactate and other monocarboxylate transmembrane transportation and on pH regulation in tumor cells.

Objective To study the mRNA expression of MCT1 and MCT2 genes in human hepatocellular carcinoma (HCC) and paracarcinoma liver tissue (PCLT). Methods The semi-quantitative analysis of MCT1 and MCT2 genes mRNA expression in human HCC and PCLT was conducted by RT-PCR method and electrophoresis band opacity density (OD) comparison analysis method in 25 patients with HCC. Results The mRNA expression of MCT1 was significantly higher than MCT2 in HCC and PCLT, in HCC the mRNA expression of MCT1 and MCT2 genes were significant higher than that in PCLT. Conclusions The high expression of mRNA of MCT1 and MCT2 genes in HCC indicates that these genes may take a significant role on lactate and other monocarboxylate transmembrane transportation and on pH regulation in tumor cells.

BACKGROUND: Dendritic cells play an important role in antigen present in vivo, and the mechanism of tumor cells in escaping the antigen presentation of dendritic cells existed in the patients with tumor.OBJECTIVE: To sensitize dendritic cells from human peripheral blood with apoptotic hepatoma cells induced by mitomycin.DESIGN: A randomized control trial by taking apoptotic hepatoma cell sensitized dendritic cells as the observed subjects.SETTINGS: Institute of Field Surgery, Daping Hospital, the Third Military Medical University of Chinese PLA; Institute of Radiation Medicine,Chinese PLA Academy of Military Medical Sciences.MATERIALS: The experiments were carried out in the Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences from April 1998 to May 1999. The cell strain was the QBC939 bile duct cancer cell strain, and mitomycin was used as the antitumor drug.METHODS: Mononuclear cells were isolated from the peripheral blood of normal people, 50μg/L granulocyte-macrophage colony stimulating factor(GM-CSF) and 1 000 U/mL interleukin-4 (IL-4) were added, once every other day for 4 times. On the 3rd day of culture, the apoptotic bile duct cancer cells induced by mitomycin was added, and then cultured in vitro for 4 days, finally the dendritic cells were collected.MAIN OUTCOME MEASURES: ① The identification of the cultured dendritic cells was observed; ② The dendritic cells were co-cultured with necrotic and normally cultured bile duct cancer cells respectively, and the phagocytized apoptotic body loaded antigens were observed; ③ The immunostimulatory activity of dendritic cells (1×103, 5×103 and 1×104/well)and that after loaded by antigen were detected, and the mononuclear cells were taken as controls.RESULTS: ① The cultured and amplified dendritic cells expressed high levels of costimulatory molecules of CD1a and B7, and there were typical irregular processes on the surface. ② The tumor cells formed apoptotic bodies when they were induced by mitomycin, which were arrested and phagocytized by dendritic cells. ③ The ability of the antigen loaded dendritic cells in stimulating the proliferation of allogenic lymphocyte T was further enhanced.CONCLUSION: The apoptotic tumor cells induced by mitomycin can induce the mononuclear cells from human peripheral blood differentiating into the dendritic cells with the concomitance of GM-CSF and recombinant IL-4 and amplify dendritic cells. Meanwhile, the dendritic cells can effectively present the antigens of apoptotic bile duct cancer cells, and it probably becomes a new effective approach for tumor antigen to sensitize dendritic cells.

Objective To explore the effect of fatty liver on graft survival, especially with reference to macrovesicular and microvesicular steatosis and to evaluate the relationship between histological grading and inflammation activity. Methods Different degrees of rat fatty liver model were established by feeding rats a diet consisting of 79% standard diet, 20% lard and 1% cholesterol. By modified two cuff vascular anastomoses and end to end suture for bile duct, rat orthotopic liver transplantation was performed to evaluate the relationship between donor histological grading and survival rate. Results Low survival rate of macrosteatosis (grade Ⅲ) was found. Most rats died of liver failure in early days after transplantation. Pathological findings showed frequent hepatic necrosis. There was no significant difference between macrosteatosia(gradeⅠ) and the normal group. After transplantation, almost all of the fat was cleared by the end of the fourth week. Diminished steatosis and liver regeneration were found in macrosteatosis (gradeⅡ), while microsteatotic donors had higher survival rate than the other groups except the normal group. Conclusion Macrovesicular steatosis(grade Ⅲ) affects graft survival and these steatotic livers should not be used as donors. However, steatotic livers with mild macrovesicular steatosis (grade Ⅰ) and microvesicular steatosis(grade Ⅲ) do not influence recipient survival, so these livers can be used safely for liver transplantation. The ischemic damage should be considered when using livers of macrovesicular steatosis(gradeⅡ). Donors with numbering score more than 2.7 are correlated with the poor survival.

Objective To determine the effect of diffusion chamber for immune isolation and the proliferation of allogenic hybridoma within diffusion chamber in mice. Methods The hybridoma cell strain was established from SP2/0 cells and the splenocytes of BALB/c mouse by PEG mediate technique. The cells were injected peritoneally into BALB/c ( n =5) and C57 BL/6 (B6, n =5) mice. Diffusion chambers with microporous membrane containing hybridoma cells were implanted peritoneally into 5 BALB/c and 5 B6 mice. Results Within two months after peritoneal injection of the cell strain, four BALB/c mice developed bloody ascites and one developed abdominal tumor, but the B6 mice developed neither ascites nor tumor. After 30 and 101 days of the chamber implantation, the chambers were coated by omentum or mesenterium in all the animals of both strain groups and freshly formed blood vessels to the chamber membrane could be observed. Within the chambers, tumor developed, but the tumor could not fully fill the inner space of the chamber during the whole observation period. Conclusion The hybridoma cells established from SP2/0 cells and the splenocytes of BALB/c mouse may carry BALB/c MHC antigen. The diffusion chambers can effectively isolate immune rejection. Freshly formed blood vessels to the chamber membrane can support the cells within the chambers.

Objective To investigate the effects of antisense TIMP 1 on the interstitial collagen synthesis of fibrotic liver in rats. Methods Fibrosis was induced in the liver of rats with the injection of 60% CCl 4 and 5% alcohol. Recombinant antisense TIMP 1 gene mammalian expression vectors were construct and infused into the fibrotic liver of rats. The expressions of TIMP 1 mRNA, ?1(Ⅲ) mRNA and protein were determined by RT PCR, ISH and immunohistochemistry. The serum levels of PCⅠ, PCⅢ in rats were determined by RIA. Results Antisense TIMP 1 gene could markedly suppress the expression of endogenous TIMP 1 mRNA in the transfected liver of rats( P

Objective To observe the survival time, pathological change and liver regeneration in different kinds of reduced size liver transplantation in steatotic rats. Methods Rat models of different kinds of reduced size orthotopic liver transplantation were performed by modified two cuff vascular anastomoses and end to end suture for bile duct to observe the recipient body weight, graft weight, recipient original liver weight, histological and pathological and electron microscopic findings in comparison with those of the whole rat liver transplantation. Results One week survival rate of the whole liver transplantation, 70% reduced size liver transplantation(ROLT), 60% ROLT and 50% ROLT group (grade Ⅰsteatotic donor) was 91.67%, 75%, 75% and 25%, respectively, and 2 week survival rate was 83.33%, 75%, 58.33% and 0, respectively. In grade Ⅱ steatotic donor, 1 week survival rate of the whole liver transplantation and 70% ROLT was 83.33% and 25%. As to donor livers with microvesicular steatosis, 1 week survival rate of the whole liver, 70% ROLT, 60%ROLT and 50% ROLT was 83.33%, 75%, 75% and 33.33% and the 2 week survival rate was 75%, 66.67% 66.67% and 0. The survival rates of 50% ROLT in grade Ⅰ steatotic donor and livers mainly with microvesicular steatosis were significantly different from those in other groups. The 1 week survival rate of 70% ROLT was very poor in steatotic donors in grade Ⅱ. Pathological findings after operation included liver regeneration and mild lymphocyte infiltration in portal space, the amelioration of the steatosis in some cases and dilation of the central vein and sinusoids. Conclusion To obtain long term survival of reduced size liver transplantation using steatotic donors, the GRBW should be over (2.28?0.12)(the ratio of graft to recipient liver weight over 60%). Steatotic livers in grade Ⅱ should not be used as donors in ROLT. The steatosis can be ameliorated after operation.

Objective:To eastablish the efficient presentation of antigen from apoptotic cells by human DC from peripheral blood. Methods: using recombinant human granulocyte/macrophage colonystimulating factor(GM- CSF) and interleukin 4 (IL- 4 ) we have established dendritic cells (DC)from peripheral blood monocyte that maintain the antigen capturing and processing capacity characteristic of immature dendritic cells in vivo. GM - CSF 50ng/ml , IL- 41 000ng/ml once two days(total four). on the 3 rd day of culture, immature DC and apoptotic hepatochlangioma cells were in coculture lasting 7 days. Results:these cells had typical dendritic morphology, express high levels of CD1a ,B7 and acquired antigen from apoptotic cells and induced an increased T cell stimulatory capacity in MLR. Conclusions:we have established DC from blood mononuclear tells using GM- CSF and IL- 4 and DC can be efficiently drived from apoptotic cells and can induce the increase of T cells obviously. It probably becomes an effective approach of antigen transduced with DC.

Objective To study the effects of high cholesterol diet and activated macrophages (M) on glycoprotein secretion from the gallbladder tissue of guniea pig.Method Forty guniea pigs were randomized into group A fed with ordinary diet and group B fed with a diet containing 1.2% cholesterol for one week.Glycoprotein secretion from guinea pig gallbladder was observed in tissue culture using ~3H-glucesamine as a precursor,and in the meantime,with hydrocortisone and activated M to understand the effects on glycoprotein synthesis and secre- tion function of gallbladder epithelium.Results The activity of peritoneal M was significantly increased in guinea pigs fed with high cholesterol diet.High cholesterol diet induced significant release of ~3H-glucosamine-labeled gly- coprotein into the tissue culture medium as compared with the control level of guinea pig fed with normal diet.The gallbladder tissues were co-cultured for 16 hours with peritoneal M of guinea pig fed with high cholesterol diet. Mucin secretion had an evident increase compared with the controls (with the peritoneal M of guinea pig fed with normal diet at 10~4,10~6 cell/ml).Hydrocortisone (10~(-6),10~(-5),10~(-4)mol/l) caused a reversible dose-dependent inhibition on glycoprotein secretion from the gallbadder tissues of guinea pig fed with high cholesterol diet.Hydro- cortisone (10~(-4)mol/l) also inhibited the stimulatory effect of M activated by high cholesterol diet on glycoprotein hypersecretion in the gallbladder tissues of guinea pig fed with ordinary diet.Conclusion (1) High cholesterol diet can induce the increase of glycoprotein secretion from gallbladder tissues of guinea pig;(2) M can be actvi- ated by high cholesterol diet,which stimulates glycoprotein secretion from the gallbladder tissues of guinea pig. Considering the results of experiment using an animal gallbladder stone model,these findings suggest that the hy- persecretion of glycuprotein from guinea pig gallbladder tissue may be related to guinea pig M activated by high cholesterol diet and stimulated to release TNF,IL-I,etc.

Objective To probe into the kinetics of gallstone formation.Methods Fifty seven rabbits were divided into five groups: (1) normal control with standard fodder, (2)1 2% cholesterol was added into the fodder,(3)1 2% cholesterol plus indomethacin in the fodder,(4)1 2% cholesterol plus erythromycin,(5) 1 2% cholesterol plus Dong Li San, a Chinese herb compound. All animals were feed four weeks before measurement.Results Gallstone developed in 0 out of 13 in group 1, in 12 out of 14 rabbits in group 2, in 4 out of 10 rabbits in group 3, in 0 out of 10 in group 4, and in 2 out of 10 in group 5. Compared with that in group 1 rabbits in group 2 had higher level of cholesterol and mucin in bile,much higher common bile duct pressure and cystic duct resistance,much lower gallbladder emptying rate ( P