Objective To investigate the prevention of HBV reinfection in the perioperative period of liver transplantation on HBV-related diseases． Methods Published papers were collected and reviewed． Results HBV-related diseases were the main indications of liver transplantation．The prevention for HBV reinfection affects the survivals remarkably. Nowadays, a lot of medication have been used in the prevention of HBV reinfection， and the therapeutic regimens were different from each other. Conclusion Liver transplantation is an effective treatment for HBV-related disease． Appropriate prevention of HBV reinfection in the perioperative period of liver transplantation is important for the survivals of patients．
Objective To determine the value of detection of micrometastasis in peripheral blood to hepatocellular carcinoma （HCC） metastasis or recurrence． Methods Reviewed the related literatures, the methods and significances of the detection of HCC micrometastasis in peripheral blood were analyzed. Results Currently, there are mainly two methods, hematogenous dissemination cell detection and HCC specific mRNA biomarker detection, for detection of HCC micrometastasis in peripheral blood. Theoretically, although they are considered as early detections of HCC metastasis or recurrence, researches still not have a abroad agreeable conclusion from different studies. After adjusting and improving the methods and detection time, different studies also have not gotten a quite consistent conclusion. Conclusion There is a great significance in detection of HCC micrometastasis in peripheral blood to understanding the mechanisms of HCC metatasis and recurrence, and also to improving the clinical therapy. Theoretically and practically, the method should be improved for facilitating the mechanism research of HCC metastasis and recurrence, and the application of detection.
ObjectiveTo investigate the expression of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) in human pancreatic adenocarcinoma and their correlation with clinicobiological behavior.MethodsThe expression of COX-2 and VEGF in 51 cases of human pancreatic ductal adenocarcinoma were detected with immunohistochemistry of Envision.ResultsExpression of COX-2 and VEGF in pancreatic ductal adenocarcinoma were 74.5% and 68.6%, respectively; no expression of COX-2 and VEGF in adjacent normal tissue was detected. Both COX-2 and VEGF expression in clinical stage Ⅲ-Ⅳ were much higher than those in clinical stage Ⅰ-Ⅱ, and also higher in positive group of lymph node metastasis than in negative group as well (Plt;0.05). None of them had relation with histological grades, age, sex, tumor size and location. The expression of COX-2 was closely correlated with VEGF (r=0.411, Plt;0.01).ConclusionCOX-2 and VEGF may play a pivotal role in tumorigenesis and tumor progression in pancreatic cancer, they may provide new targets for therapy of pancreatic cancer.
Objective To construct a green fluorescent protein expression plasmid pEGFP-C3-anti-TAG72 scFv-CD28, containing anti-TAG72 single chain variable fragment (scFv) fused into the transmembrane and intracellular domain of the signal-transducing chain of CD28 gene, and to transfect it into peripheral blood mononuclear cells． Methods Recombinant transmembrane and intracellular domain of CD28 cDNA and anti-TAG72 scFv cDNA fragment was subcloned into pEGFP-C3 vector. Recombinant clones were selected by Kanamyein， and then identified by PCR， enzyme digestion analysis and DNA sequencing. The recombinant plasmid was transfected into peripheral blood mononuclear cells by means of lipofection. The recombinant protein expression was confirmed by immunocytochemistry, laser scanning confocal microscope, PCR and Western blot analysis. Results The fused gene fragment of anti-TAG72 scFv-CD28 was successfully inserted into pEGFP-C3 plasmid, and it was confirmed by enzyme digestion and DNA sequencing. The fused anti-TAG72 scFv-CD28 gene and its protein was identified in peripheral blood mononuclear cells． Conclusion The eukaryotic expression plasmid pEGFP-C3-anti-TAG72 scFv-CD28 was successfully constructed and transiently expressed in peripheral blood mononuclear cells， which would lay a foundation for further studies on the role of it to activate tumor-associated antigen-specific T lymphocyte, for generating of modified T lymphocytes targeting gastrointestinal tumors.
Objective To study the prokaryotic expression of the anti-tumor associated glycoprotein-72 (TAG-72) singlechain fragment variable (scFv) antibody and its specific affinity to hepatocellular carcinoma cell lines and tissues. Methods The cDNA of anti-TAG-72 scFv antibody was inserted into pCANTAB5E to obtain phage vector anti-TAG-72 -scFv-pCANTAB5E. Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of anti-TAG-72 scFv antibody. SDS-PAGE and Western blot were used to identify the anti-TAG-72 scFv antibody. Human hepatocellular carcinoma cells were cultured, and TAG-72 was determined with the obtained scFv by immunohistochemistry in the cells and paraffin-embedded hepatocellular carcinoma tissues. Results SDS-PAGE and Western blot showed that the anti-TAG-72-scFv antibody was successfully expressed. Anti-TAG-72 scFv could bind to hepatocellular carcinoma cell lines SMMC7721, HepG2, and HHCC, but not to BEL7402, suggesting that SMMC7721, HepG2, and HHCC cells expressed TAG-72. For the 40 cases of hepatocellular carcinoma tissues, the positive rate of TAG-72 in stage Ⅰ and Ⅱ-Ⅲ was 23.08% (3/13) and 62.96%(17/27), respectively. While no TAG-72 expression was found in the 10 normal cases. TAG-72 expression was significantly different between hepatocellular carcinomas and normal tissues (Plt;0.05). Conclusions The prokaryotic expression of anti-TAG-72 scFv antibody is successfully achieved, and can be used to identify TAG-72 antigen. TAG-72 is highly expressed in hepatocellular carcinoma, but not in normal liver tissue, which may be suggested as cancer marker of hepatocellular carcinoma.
Objective To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of αfetoprotein （AFP） enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection. Methods Angiostatin cDNA was amplified from normal human eukaryotic cells by using RTPCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1AFABangiostatin K5His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with antiHis antibody. Results The 500base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1AFABangiostatin K5His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDSPAGE and Western blot. Conclusion Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.
Objective To explore the expression and function of NDRG2 gene in human primary hepatocellular carcinoma and normal hepatic tissues. Methods The immunohistochemical ABC method， Western blot， and Real-time PCR were used to investigate the expression and content of NDRG2 in human hepatocellular carcinoma and hepatic normal biopsies. Results The NDRG2 protein located in cytoplasm. The positive rate was 16.67%（5/30） and 100%（30/30） in hepatocellular carcinoma and normal hepatic tissues, respectively. The relative content of NDRG2 protein in hepatocellular carcinoma and normal hepatic tissues were 0.029 0±0.005 9 and 0.109 2±0.002 8. There were significant differences between human hepatocellular carcinoma and hepatic normal biopsies both in staining positive rates and relative content（P＜0.05）. The Western blot also agreed with the result，the expression level of NDRG2 protein in hepatocellular carcinoma and normal hepatic tissues was 1.13±0.15 and 1.57±0.18， respectively, there was significant difference（P＜0.05）. Also, compared with normal hepatic tissues, the expression level of NDRG2 mRNA in carcinoma tissues was reduced significantly (0.89±0.15 vs. 1.48±0.17, P＜0.05). However, there were no significant differences in NDRG2mRNA expression between Edmondson-Steiner grades. Conclusions There possibly have difference in NDRG2 expression between human primary hepatocellular carcinoma and normal hepatic tissue. NDRG2 gene may take part in the pathogenesis of human primary hepatocellular carcinoma. Futher study will be needed to study its mechanism and function.
Objective To explore the differential expressions of seven microRNAs between hepatocellular carcinoma (HCC) and adjacent nontumorous tissues (NT), analyze the correlations between differential expressing microRNAs and the levels of tumor markers in serum, and furnish evidence for novel diagnostic and prognostic tool of HCC. Methods Real-time quantitative PCR technique was used to measure the differential expressions of seven microRNAs in HCC tissues compared with NT. Results Compared with NT, the relative expressions of seven microRNAs in HCC tissues manifested statistical difference (Plt;0.05). MiR-34c, miR-21, miR-16, and miR-10b presented higher expressions in the HCC samples than those in the NT samples, while miR-200a, miR-148b, and miR-Let-7i demonstrated lower expressions in the HCC samples than those in the NT samples. In addition, miR-200a and miR-148b were markedly down-regulated in the HCC tissues than those in the NT. The differential expressions of miR-200a in HCC compared with NT samples was correlated with serum AFP level of the patients (r=0.848 9, Plt;0.01), while the differential expressions of the other six microRNAs had no correlation with the levels of tumor markers in serum (Pgt;0.05). Conclusions There are differential expressions of microRNAs between HCC and NT. MiR-200a may serve as a novel diagnostic and prognostic tool of HCC.
ObjectiveTo discuss the effect of palliative drainage operation on the life quality of hilar cholangiocarcinoma. MethodsCholangiocarcinoma data of our hospital in recent 21 years were analysed retrospectively. They were divided into four groups: RouxenY choledochojejunostomy group, bridge internal drainage group, PTCD (or ERBD) internal drainage group, and operative external drainage group. The operative mortality, incidence of postoperative cholangitis and survival period were compared among groups.ResultsThe total perioperative mortality of 193 cases of palliative operation was 9.3%, there was no difference among groups (P＞0.05). The rate of postoperative cholangitis in the bridge internal drainage group (10.0%) was lower than that of RouxenY choledochojejunostomy group (19.4%),P＜0.05, the rate of cholangitis in PTCD (or ERBD) internal drainage group (37.5%) and operative external drainage group (38.1%) were significantly higher than that of RouxenY choledochojejunostomy group (P＜0.01). There was no significant difference between RouxenY choledochojejunostomy group 〔(9.2±1.8) months〕 and PTCD (or ERBD) internal drainage group 〔(8.8±1.9) months〕 in survival period (P＞0.05)，but the survival period of the above groups were significantly higher than that of bridge internal drainage group 〔(6.5±1.6) months〕，P＜0.05, and operative (or PTCD) external drainage group 〔(4.3±2.0) months〕,P＜0.01.ConclusionThe life quality of RouxenY choledochojejunostomy group is better than that of bridge internal drainage group and PTCD (or ERBD) internal drainage group, the life quality of external drainage is worse than that of the other groups.
Objective To investigate the expression of RNA editase ADAR1 in the lymphocytes in rats’ spleen with liver transplantation rejection. Methods Thirty SD rats and 75 Wistar rats were included. Fifteen livers from Wistar rats were transplanted to 15 Wistar rats (isograft group), 30 livers from SD rats were transplanted to 30 Wistar rats (allograft group and allograft+FK506 group), and 15 of them were then intramuscularly injected with FK506, 2 mg/(kg·d), the other 15 Wistar rats were only operated similarly to the other rats without any liver transplantation (control group). Five rats were killed and their splenetic tissues were collected on day 3, day 5, and day 7, respectively. The expression of ADAR1 mRNA in lymphocytes of the spleen in acute rejection was detected by RT-PCR. Results Different performance of pathology was observed in all the liver and spleen tissues from the transplanted rats over time, especially in allograft group. The expression of ADAR1 mRNA in the allograft group was significant higher than that of isograft and allograft+FK506 groups （P＜0.001）, especially on the 5th day. Conclusion There was a significant positive correlation between expression of ADAR1 and the severity of acute rejection, but the mechanism by which ADAR1 affected the acute rejection is unknown and needs to be further studied. FK506 may inhibit the expression of ADAR1 and remarkably reduce the severity of acute rejection.