ObjectiveTo study the relationship between hepatocellular apoptosis and glycogen contents during hepatic cold preservationreperfusion and its mechanism.MethodsBased on the model of four groups of rabbit livers with different hepatocellular glycogen contents, hepatocellular apoptosis and bax gene expression were observed during hepatic cold preservationreperfusion.ResultsApoptotic hepatocytes were obviously found in 60 minute reperfusing livers subsequent to 9 hour cold storage, and there was significant difference in the numbers of apoptotic hepatocytes among all the groups. In the same time, there was the close relationship between the levels of bax gene expression and the glycogen contents of hepatocytes.ConclusionIntracellular abundant glycogen may significantly depress the hepatocellular apoptosis during hepatic cold preservationreperfusion by decreasing hepatocellular bax gene expression.
ObjectiveTo study the expression of survivin protein in primary hepatocellular carcinoma(PHC) and its relationship to the proliferation of the tumor cells and prognosis of PHC. MethodsThe expression of survivin protein and the proliferation of tumor cells marked by proliferating cell nuclear antigen (PCNA) in 48 cases of PHC were determined by immunohistochemical method. ResultsThe survivin protein was expressed in 31 of 48 cases of PHC (64.6%). The expression of PCNA was significantly higher in hepatocellular carcinoma (HCC) with positive survivin expression than in HCC with negative survivin expression. The patients with positive survivin expression had the worse prognosis than those with negative survivin expression. ConclusionThe expression of survivin may play an important role in the proliferation of PHC cells and closely associate with the prognosis of PHC, and probably become the prognostic factor and an important target of therapy.
Objective To prepare the immunonanospheres［SC3Ab-HSA(5-Fu)-NS］ against human colorectal cancer and evaluate its immunoreactivity and effects on cancer. Methods SC3Ab-HSA(5-Fu)-NS was prepared by intermolecular cross-linking the monoclonal antibody SC3Ab with human serum albumin nanospheres containing 5-Fu ［HAS(5-Fu)-NS］ via new hetero-bifunctional crosslinker SPDP. Condensation test and immunoflurecence were used to evaluate the immunoreactivity, the specific binding of SC3Ab-HSA(5-Fu)-NS with colorectal cancer cell line SW1116 was observed by microscope and electron microscope. The specific cytotoxic effects on target cells were evaluated in vitro by MTT assay. SC3AbHSA(5-Fu)-NS, HSA(5-Fu)-NS and 5-Fu were injected into nude mice bearing human colorectal carcinoma, to study the inhibitory activity of SC3Ab-HSA(5-Fu)-NS in vivo. Results The immunoreactivity of SC3Ab-HSA(5-Fu)-NS was well preserved. SC3Ab-HSA(5-Fu)-NS can bind the SW1116 cells specifically. The IC50 value for SC3Ab-HSA(5-Fu)-NS on SW1116 cells was 24.6 μg/ml,which was lower than that of HSA(5-Fu)-NS(345.3 μg/ml) and 5-Fu(325.6 μg/ml). The inhibitory rate of SC3Ab-HSA(5-Fu)-NS on the growth of colorectal cancer xenografts was significantly higher than that of HSA(5-Fu)-NS or 5-Fu(P＜0.001).Conclusion SC3Ab-HSA(5-Fu)-NS has immunoreactivity and specific active targeting to the colorectal cancer cells. The anticancer ability of SC3Ab-HSA(5-Fu)-NS is significantly higher than that of HSA(5-Fu)-NS and 5-Fu.
Objective To study the results of in situ microwave thermocoagulation therapy for liver neoplasms. Methods Thirty-one patients (male 28, female 3) with liver neoplasms underwent in situ microwave thermocoagulation therapy in recent 4 years were studied. The time of the therapy arranged from 2 to 6 minutes with the core temperature from 110℃ to 125℃. Twenty six of the thirty one (83.9%) were followed up. Results Ninty point three percent of these patients have a good result. The average survival time after the operation was 19.7 months. One-year and three-year survival rate were 77.4% and 38.7%, respectively. Conclusion The in situ microwave thermocoagulation therapy have the advantages of causing slight trauma, promoting repair, good tolerance and curative effectiveness. It’s a simple, safe and effective method with less adverse effect for treating the liver neoplasms, especially for unresectable neoplasms.
Objective To discuss the way of animal model building of hepaticocholedochostomy(HC) and hepaticojejunostomy(HJ) and to compare the short-term effect. Metheds Twenty-nine dogs were divided randomly into control group(n=5) and the experimental group (stenosis of left hepatic duct, n=24). After 7 weeksof stenosis of left hepatic duct,24 dogs in the experimental group were divided randomly into HC subgroup (n=12) and HJ subgroup (n=12) .The operation time and the blood loss during operation were recorded and the hepatic function was detected.Results The diameter of left hepatic duct was significantly expended after 7 week’s stenosis. Hepaticocholedochostomy took shorter time and lost less blood than hepaticojejunostomy. The dogs in HC subgroup lost less weight than thosein HJ subgroup. In HC and HJ subgroups, the mortality rates were 1/12 and 3/12;the infectious rates of incision were 3/12and 5/12 respectively. Serum levels of total bilirubin and transaminase increased significantly in the 7th week after stenosis of left hepatic duct compared with before stenosis of left hepatic duct. However, Serum levels of total bilirubin and transaminase restored to normallevels after 1 month of HC or HJ.Conclusion It is feasible to establish animal model of bile duct reconstruction on the basis of stricture of bile duct. The dogs undergoing hepaticocholedochostomy have less trauma, better results than the dogs undergoing hepaticojejunostomy. Both hepaticocholedochostomy and hepaticojejunostomy are able to relieve the obstruction of bile duct.
Objective To find the relation between donor liver cold preservation-reperfusion injury and hepatocellular apoptosis during liver transplantation. Methods Four groups of rabbit livers, which had experienced cold storage for 0,3,6,9hr respectively, were observed during their cold preservation-reperfusion by using TdT-mediated dUTP-biotion nickend labeling and electron microscope.Results Apoptopic hepatic parenchymal cells were obviously observed in reperfusing livers subsequent to cold storage. Furthermore, the longer the cold storage duration, the greater the number of apoptotic cells. On the contrary, no or rare apoptotic hepatic parenchymal cells was observed in all the groups at the end of cold preservation. Conclusion It suggests that apoptosis of hepatic parenchymal cells is markedly involved in donor liver cold preservation-reperfusion injury.
Objective To study the protective effects of pre-storing glycogen on warm ischemia reperfusion injury during partial hepatectomy. Methods Thirty-eight patients were randomly divided into a trial group (n=19) and a control group (n=19). In the trial group, patients were given high concentration glucose intravenously during the 24 hours before the operation. The hepatic lesion was resected after portal triad clamping in the two groups. Liver function of all patients was measured before the operation and the first and fifth days after the operation. Normal hepatic tissue was biopsied to measured hepatic tissue glycogen contents before the operation and the change of superoxide dismutase (SOD) at the point of pre-ischemia, post-ischemia, and reperfusion 2 hour. Bcl-2 mRNA, a well known anti-apoptotic factor, was also detected using quantitative polymerase chain reaction. Results The hepatic tissue glycogen content of the trial group was significantly higher than that of the control group before the operation (Plt;0.01). Liver function of the trial group was significantly better than that of the control group on the first and fifth day after operation (Plt;0.05). There was significant difference in SOD activity between the two groups at the end of hepatic vascular occlusion and at the point of 2-hour reperfusion (Plt;0.05). Furthermore Bcl-2 mRNA expression of the trial group was notably up-regulated at the point of 2-hour reperfusion compared to the control group. Conclusion Pre-store storing glycogen might protect liver ischemia reperfusion injury caused by hepatic vascular occlusion during partial hepatectomy. The potential mechanism might be that pre-storing glycogen enhances Bcl-2 expression.
Objective To detect the anti-colon cancer ability of whole cell lysates pulsed dendritic cell (DC) which acts as an adjuvant. Methods Whole cell lysates of LoVo cells were extracted with freeze thawing method, then the monocyte-derived DC were pulsed with this cellular antigen. Subsequently, the capability of antigen pulsed DC to promote T lymphocytes proliferation and the ability of T lymphocytes to kill LoVo cells were detected by 3H-TdR incorporation and lactate dehydrogenase release methods, respectively. Results The whole cell lysates of LoVo cells pulsed DC significantly stimulated T cells proliferation, and the cytotoxicity abilities of primed T cells to kill LoVo cells were also enhanced. Conclusion Tumor cell lysates which act as the cellular antigen to pulse DC can improve the efficacy of anti-cancer immune response， which provides us an experimental evidence for cancer immunotherapy.
Objective To construct AWP1 (associated with protein kinase C related kinase 1) recombinant adenovirus as the tool of transferring the gene and investigate its expression and localization in human vascular endothelial cell ECV304. Methods Cloned AWP1 cDNA was inserted into the multiply clone sites (MCS) of plasmid pcDNA3 for adding flag tag, and the flag-AWP1 gene was subcloned into shuttle vector pAdTrack-CMV. After identified with restrictional enzymes, plasmid pAdTrack-flag-AWP1 was linearized by digestion with restriction endonuclease PmeⅠ, and subsequently cotransformed into E.coli BJ5183 cells with adenoviral backbone plasmid pAdEasy-1 to make homologous recombination. After linearized by PacⅠ, the homologous recombinant adenovirus plasmid transfected into 293 cells with Lipofectamine to pack recombinant adenovirus. After PCR assay of recombinant adenovirus granules, recombinant adenoviruses infected 293 cells repeatedly for obtaining the high-level adenoviruses solution. And then, the recombinant adenoviruses infected human ECV304 cells for observing the expression and localization of AWP1 under laser scanning confocal microscope (LSCM). Results PCR assay showed that recombinant adenovirus Ad-flag-AWP1 was obtained successfully; and ECV304 cells were infected high-efficiently by the homologous recombinant virus. Then, it was observed that flag-AWP1 protein expressed in ECV304 cells and distributed in the leading edges of the cell membrane. Conclusion The vectors of flag-AWP1 recombinant adenovirus are constructed, and the localization of AWP1 protein in ECV304 cells might show that AWP1 may be a potential role on the cell signal transduction.
ObjectiveTo explore the regulation of nuclear factorκB (NFκB) on tumor necrosis factorα (TNFα) expression in the liver and its role in liver injury in rats with acute pancreatitis.MethodsSeventytwo Wistar rats were randomly divided into three groups: acute pancreatitis group (AP), acute pancreatitis treated with pyrrolidine dithiocarbamate (PDTC) group (APP) and sham operation group (SO). The hepatic NFκB activities were determined with electrophoretic mobility shift assays. The expressions of hepatic TNFα mRNA were detected with RTPCR. The levels of serum alanine aminotransferase (ALT) were also measured.ResultsThe NFκB activities were significantly higher in AP and APP groups than those in SO group 3-6 hours after operation. The expressions of TNFα mRNA were ber in AP and APP groups than those in SO group 3-24 hours after operation. The levels of serum ALT were also significantly higher in these two groups than those in SO group 3-24 hours after operation. However, compared with AP group, the activities of NFκB, the expressions of TNFα mRNA and the levels of ALT significantly decreased in APP group.ConclusionThe activation of hepatic NFκB is associated with the liver injury by regulating TNFα mRNA expression in acute pancreatitis.