【Abstract】ObjectiveTo inquire the therapeutic effect of retroperitoneal splenic autotransplantation combined with lower esophageal transection in the treatment of hepatic cirrhosis induced portal hypertension with randomized comparasion.MethodsThe hepatic cirrhosis induced portal hypertension patients with Child A or B grade of liver function were randomly divided into splenic autotransplantation group and splenectomy group.In the splenic autotransplantation group, retroperitoneal transplantation of pedicled autosplenic tissue combined with modified lower esophageal transection was performed,while in the splenectomy group, splenectomy combined with modified lower esophageal transection was conducted.The general conduction, splenic scanning, liver function, serum tuftsin and IgM levels in patients were observed 2 to 6 months after operation, and compared with those before operation. ResultsOne patient died in the splenectomy group on the 6th postoperative day, rebleeding occurred in one case of the splenic autotransplantation group. The levels of tuftsin and IgM in splenic autotransplantation group were higher than those of splenectomy group after operation, with significant difference (P＜0.01). The liver function between two groups showed no difference (Pgt;0.05).ConclusionSpleen autografts could maintain the basic immune function of spleen and survive for a long time.
【Abstract】Objective To evaluate the value of pTNM classification in predicting the prognosis of hepatic cell carcinoma after liver transplantation. Methods Fifty-nine HCC cases undergoing liver transplantation between April 1993 and January 2003 were retrospectively reviewed. Fiftynine cases were staged by using the pTNM classification. Results The 1-year survival rates were 66.67%, 66.67%, 40.91% and 31.75% for Ⅰ，Ⅱ，Ⅲa and Ⅳa stages，2-year survival rates were 66.67%, 66.67%, 21.29% and 31.75%, the difference was not statistically significant. Conclusion The pTNM classification is not good enough to predict the prognosis of hepatic cell carcinoma after liver transplantation.
Objective To investigate the expression of human leukocyte antigen-G （HLA-G） in hepatocellular carcinoma （HCC） tissues， and to evaluate the prognosis of patients after liver transplantation. Methods The clinical data of 83 patients with HCC who underwent liver transplantation from January 2004 to May 2008 in the Liver Transplantation Center of the Third Affiliated Hospital of Sun Yat-sen University were analyzed retrospectively. The expression of HLA-G in HCC tissues was detected by using immunohistochemical analysis. The Kaplan-Meier method was used to evaluate the cumulative survival rate and tumor-free survival rate. Log-rank test and Cox regression model were used to analyze the single and muti-factor for tumor-free survival rate, respectively. Results Among the 83 patients，there were tumor recurrence in 35 patients （42.2%）. The 1-year，3-year, and 5-year of cumulative survival rate was 97.2%，89.8%, and 43.1%, respectively. The 1-year，3-year， and 5-year of tumor-free survival rate was 93.6%，68.9%, and 38.7%, respectively. The positive rate （68.7%） of HLA-G expression in HCC tissues was significantly higher than that in paracancerous tissues （15.7%）, P＜0.01. A significant association was found between expression of HLA-G and tumor size， vascular invasion, and pathology differentiation （P＜0.05）. Single factor analysis showed that the expression of HLA-G （P＜0.01）， tumor size （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.01） effected on tumor-free survival rates of HCC patients after liver transplantation. The tumor-free survival rate in positive expression of HLA-G group was significantly lower than that in negative expression of HLA-G group （P＜0.01）. Cox regression model analysis showed that the expression of HLA-G （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.05） were independent risk factors that affected the tumor-free survival rate of HCC patients after liver transplantation. Conclusions There is expression of HLA-G in HCC tissues. The independent risk factors that affecting the tumor-free survival rate of HCC patients after liver transplantation include the expression of HLA-G， vascular invasion, and pathological differentiation. Taking interferential measures for patients with positive expression of HLA-G and strict selection of indication of liver transplantation for HCC can reduce the recurrence rate of tumor.
Objective To construct the expression vector of HLA-G-shRNA and investigate the effect of HLA-GshRNA from NK cell lysis. Methods Four HLA-G shRNA plasmids were constructed and transiently transfected to Bel-7402 cell lines， the levels of mRNA and protein of HLA-G were detected by Real-Time PCR and Western blot. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay. Results The gel electrophoresis and sequencing showed that the inserted sequence was identical to the one which we designed, and no aberrations such as mutation，deletion or insertion occurred. The expressions of HLA-G confirmed by Real Time-PCR and Western blot were significantly down-regulated. Bel-7402 cell lines transfected HLA-G shRNA showed higher lytic activity （P＜0.01）. After KIR2DL4 receptor blocked，lytic activity of NK-92 MI cell were decreased （P＜0.01）. Conclusions HLA-G shRNA plasmids are successfully constructed and HLA-G down-regulated can increase NK cytolysis against Bel-7402 cell. After HLA-G combines with KIR2DL4 receptor at the surface of NK cells， the inhibition effect is transferred.
Objective To study the effect of anti-CD40L monoclonal antibody on the rejection of rat pancreatic islet xenografts and its mechanism. Methods The animal models of human-rat pancreatic islet xenografts were established and were treated with anti-CD40L monoclonal antibody. The levels of blood glucose of transplantation rats were measured and the survival of grafts and transplantation rats were observed after transplantation. The morphological changes of grafts were observed and the levels of cytokines (IL-2 and TNF-α) were quantified by ELISA. Results ①Level of blood glucose in all the rats with diabetes decreased to normal on day (2.3±0.2) after transplantation. The average level blood glucose of control group began to increase on day (8.1±0.6), while the treatment group began to increase on day (18.5±1.2) after transplantation, which was significantly postponed compared with control respectively (P＜0.01). ②Grafts of treatment group and control group survived for (22±8.2) and (10±2.1) days respectively. Survival of grafts in treatment group was significant longer than that in control group (P＜0.01). ③Survival of transplantation rats were (35±6.5) and (21±5.7) days in treatment group and control group respectively. The survival of transplantation rats in treatment group was significant longer than that in control group (P＜0.05). ④Levels of serum IL-2 and TNF-α in control group increased dramatically within (3.2±0.3) days and reached peak within (7.3±0.5) days after transplantation, which were significantly higher than those measured before transplantation (P＜0.01); While in treatment group, the levels of serum IL-2 and TNF-α began to increase on day (22.6±1.7) after transplantation, and reached peak on day (28.5±2.2), which was significantly postponed than those in control group (P＜0.01). Conclusion Anti-CD40L monoclonal antibody can inhibit the rejection of rat pancreatic islet xenografts and prolong the survival time of transplantation rats and grafts.
Objective To investigate the cell growth inhibition and apoptosis induced by rapamycin on human hepatocellular carcinoma Bel-7402 cells and to study the role of mitochondrium membrane potential in the process of apoptosis. Methods Bel-7402 cells in vitro were given 5, 10, 20, 30, 40 and 50 nmol/L different concentrations of rapamycin, and the cell growth inhibiting ratio of Bel-7402 was assessed by MTT assay. The changes of morphology of Bel-7402 were observed by Hoechst 33258 staining and flow cytometry (FCM), respectively; The cell mitochondrial membrane potential was detected by using JC-1 staining method. Results Rapamycin could inhibit the growth of Bel-7402 cells significantly by inducing apoptosis, and the growth suppression and the cell apoptosis both presented time-effect relationship and were also dose-dependent. The rates of inhibiting and cell apoptosis after 72 h exposure to 50 nmol/L rapamycin were significantly higher that those of other groups (P＜0.01). Typical morphological changes of cell apoptosis were observed very clearly after the Bel-7402 cells had been exposed to rapamycin for 48 hours using Hoechst 33258 staining method, and it was also observed that the mitochondrial membrane potential decreased when apoptosis occured (P＜0.01). Conclusion Rapamycin could inhibit the growth of Bel-7402 cells by inducing cell apoptosis, and the descent of mitochondrial membrane potential may play an important role in the process of cell apoptosis.
Objective To study the superiority and efficiency of total peritoneum intraperitoneal onlay mesh (TPIPOM) in laparoscopic inguinal hernioplasty. Methods One hundred and five cases of laparoscopic inguinal hernioplasty with TPIPOM and 34 cases of inguinal hernioplasty with trans-abdominal preperitoneal laparoscopic mesh repair (TAPP) were performed from January 2002 to August 2005. Perioperative data and follow-up results were collected and compared in two groups. Results The laparoscopic hernioplasty was successfully performed in all patients. The total operation time, hospital stay, average off-bed time, duration of pain in TPIPOM group were significant shorter than those in TAPP group 〔(30.8±10.3) min vs (68.4±22.4) min, (3.8±1.3) d vs (4.3±1.5) d, (1.2±0.5) d vs (1.8±0.7) d, (1.0±0.5) d vs (1.6±0.9) d, respectively〕, P＜0.01, the total hospital cost was RMB 5 000.8±800.5 in TPIPOM group and that was RMB 8 000.5±950.6 in TAPP group (P＜0.01). No significant scrotal edema was observed postoperatively and no recurrence reported during (18.6±8.9) months follow-up in both groups. Conclusion Laparoscopic inguinal hernioplasty with TPIPOM is safe and efficacy with advantages of mini-invasion, simple procedures, shorter operation time, no complications and better recovery.
【Abstract】ObjectiveTo investigate the growth effect of methylprednisolone （MP） on human hepatocellular carcinoma cell line HepG2.Methods Periodic distribution of cells and cellular apoptosis were detected by using cell culture,immunofluorescence staning of Annex Ⅴ and flow cytometric analysis in hepatocellular carcinoma cell.Results Compared with control group, methylprednisolone increased Ｇ0/Ｇ1 phase cell, decreased S phase cell on human hepatocellular carcinoma cell line HepG2 ,which had positive correlation with the time.The apoptosis rate and the necrosis rate of cells had the relation of dose-dependent with the concentration of MP, the cell membrane of early cellular apoptosis was stained green fluorescence. Conclusion Methylprednisolone can induce Ｇ0/Ｇ1 arrest , may play a proliferation-inhibition effect on the hepatocellular carcinoma cell line HepG2.