Construction and application of a noval gene-target therapy system in hepatocellular carcinoma / 生物医学工程学杂志

This paper is aimed to explore the efficiency of a noval gene-target therapy system in hepatocellular carcinoma (HCC) treatment in vitro, by using survivin gene as the target. A new fusion promoter (AV) driving pcDNA3. 1 (-)AV plasmid, which contained the cytomegalovirus (CMV) enhancer and alpha-fetoprotein (AFP) promoter, was constructed by molecular biologic method. The eukaryotic expression plasmid pcDNA3. 1(-)AVGFP and pcDNA3. 1 (-)AVsiRNA-survivin were constructed by cloning and inserting the green fluorescent protein (GFP) and siRNA-survivin sequence into pcDNA3. 1(-) AV plasmid separately. Then these two plasmids were transfected into HepG2, SMMC-7721 and Hela cells by using nanoparticles of calcium phosphate. The transfection efficiencies were detected by GFP. Reversed transcript polymerase chain reaction (RT-PCR) and western-blot were used to evaluate the knockdown efficiency of siRNA-survivin. The growth curves and cell death of HepG2 cells transfected with or without pcDNA3. 1(-)AVsiRNA-survivin were detected by MTT assay and flow cytometry assay, respectively. The results showed that after transfected with pcDNA3. 1(-)AVGFP vector, only HepG2 cells displayed strong GFP signaling, whereas, no GFP was found in Hela cells, suggesting that AV promoter can specifically drive downstream of gene expression in HCC cells. Furthermore, the mRNA and protein expression levels of survivin in HepG2 cells, but not in Hela and SMMC-7721 cells, were significantly silenced after pcDNA3. 1(-)AVsiRNA-survivin transfection. Finally, compared with the control cells, HepG2 cells, which were transfected with pcDNA3. 1(-) AVsiRNA-survivin plasmid, presented 68.8% cell death, including 38.68% apoptosis and 30.12% necrosis, and significant cell growth inhibition (P < 0.05). These findings indicated that this noval gene-target therapy system could specifically target HCC cells with high efficiency, providing a new gene therapy strategy for HCC.

MAGI1 inhibits cancer cell migration and invasion of hepatocellular carcinoma via regulating PTEN / 中南大学学报(医学版)

Objective To explore the biological function and molecular mechanism of membrane associated guanylate kinase,WW and PDZ domain containing 1 (MAGI1) in hepatocellular carcinoma.Methods HepG2MAGI1 stable cell line was constructed by transfecting HepG2 cells with pcDNA3.1-MAGI1 plasmid.Wound healing and invasion assay were performed to compare the migration and invasion ability of HepG2MAGI1 and HepG2 cells.Furthermore,the expression of MAGI1 and phosphatase and tensin homolog deleted on chromosome ten (PTEN) was also examined by Western blot and the relationship was analyzed.Results The wound healing assay showed that the closure of HepG2MAGI1 cells was significantly slower than that of HepG2 cells [(90±10)% vs.(50±15)%,P＜0.05],and the invasion assay showed that the number of HepG2MAGI1 cells that passed through the matrigel was fewer than HepG2 cells (68±18 vs.150±30,P＜0.05).The protein expression level of PTEN was significantly elevated in HepG2MAGI1 cells compared with HepG2 cells (1.40±0.32 vs.0.28±0.15,P＜0.05).MAGI1 and PTEN protein expression levels were positively correlated (r=0.913,P＜0.01).Conclusion MAGI1 may inhibit the cancer cell migration and invasion of hepatocellular carcinoma via regulating PTEN.

RNA interference of HDAC7 expression in hepatocellular carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To investigate the role of histone deacetylase 7 (HDAC7) in the occurrence and development of hepatocellular carcinoma.@*METHODS@#HepG2 cells and human microvascular endothelial cells (HMEC-1) were divided into 3 groups after transfection pSUPER-HDAC7 retroviral interference plasmid: a pSUPER(HDAC7RNAi+) group, a pSUPER(HDAC7RNAi-) group, and a pSUPER group as the control group. The expression of HDAC7, p21, cyclin E, matrix metalloproteinases 10 (MMP10), and hypoxiainducible factor-1alpha (HIF-1alpha) were detected by Western blot. The expression of HDAC7 in cell lines was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry, the nude mice modle, and vascular endothelial cells 2-dimensional tubulogenesis in vitro and in vivo.@*RESULTS@#Compared with the control group and the pSUPER(HDAC7RNAi-) group, the expression of HDAC7 was downregulated, the rate of cell growth inhibition and apoptosis in the pSUPER(HDAC7RNAi+) group increased more significantly; the expression of p21 and HIF-1alpha was increased significantly, while the expression of cyclin E and MMP10 in the pSUPER(HDAC7RNAi+) group was downregulated (P<0.05).@*CONCLUSION@#The expression of HDAC7 protein plays an important role in the apoptosis and vascular tubulogenesis of hepatocellular carcinoma by the upregulation of p21 and HIF-1alpha and the downregulation of cyclin E and MMP10.

Analysis of the causes and complications of reoperation for thyroid diseases / 中国普通外科杂志

Objective To study the causes of thyroid reoperation and the prevention of operative (complications). Methods The clinical data of 96 cases who underwent reoperation of thyroid were (retrospctively) analysed. Results There were 31 cases of thyroid carcinoma(14.0%, P0.05). Conclusions Thyroid (reoperation) is more difficult than primary operation, and has a higher post-operative complications rate. (Thyroid) reoperation should be avoided or decreased, so the operator should do the best at the time of the (primary) operation, and timely reoperation should be performed when necessary. Thyroid reoperation should be performed carefully to decrease complications as much as possible.

Diagnosis and treatment of hashimoto′s disease complicated with thyroid adenomas / 中国普通外科杂志

Objective To investigate the diagnosis and treatment of hashimoto′s disease(HD) complicated with thyroid adenomas(TA).Methods Clinical data of 50 cases of HD complicated with TA were analysed retrospectively. Results 8 patients were diagnosed before operation ,misdiagnosis rate was 84.0%.Intra operation frozen section diagnosis rate was 81.4% (35/43).All patients were diagnosed as HD complicated with TA by pathology after operation.Various extension thyroidectomies were performed according to the patients conditions. After operation,27 cases were given small dosage of thyroxine and 2 cases were given small dosage of prednisone. All the 50 patients were followed up for 3 months to 1 year, none presented hypothyroidism.Conclusions Complete serum immunologic examinations,ultrasonography and fine needle aspiration biopsy are helpful for the diagnosis of HD with TA. HD with TA should be treated surgically,and the intraoperation frozen section for the diagnosis is important.The operation procedures,extension of excision and medication after operation should be individulization to avoid the occurrence of postoperative hypothyroidism as possible.

Research on the expression difference of hot shock protein 70 before and after laparoscopic cholecystectomy for acute cholecystitis patients / 中国普通外科杂志

Objective To compare the expression difference of heat shock protein 70 (HSP70)!in blood of acute cholecystitis (AC) patients before and after laparoscopic cholecystectomy, and to explore the relationship between HSP70 and AC at molecular and proteinic level.Methods Subjects in the study included 85 in-hospital AC patients who received laparoscopic cholectectomy. Blood before and after laproscopic operation were obtained. Ezyme linked immunosorbent assay (ELISA) was used to determine the quantity of HSP70. Four subjects were randomly selected and the total RNA and protein in blood were extracted. RT-PCR and Western-blotting were applied to determine the expression of HSP70. The data on the figure was analysed with the software Quantity one of Bio-rad Company.Results The expression level of HSP70 of acute cholecystitis patients before laparoscopic cholecystectomy (181.42?13.58) was significantly higher than that of those patients after laparoscopic cholecystectomy (63.74?9.03) (P

Prognostic risk factors and therapeutic effect of radiofrequency ablation for primary hepatic carcinoma:a report of 195 cases / 中国普通外科杂志

Objective To investigate the effect and the prognostic factors of radiofrequency ablation(RFA) for primary liver cancer(PLC).Methods RFA was performed in 195 patients with PLC,and the efficency was evaluated with univariate analysis.Cox′s regression analysis model was used to analyze the factors affecting the prognosis.Results In the whole group of patients,the 1,2,3 and 5-year overall survival rate was 80.5%,67.4%,49.1% and 32.7% respectively,and in the patients with small PLC(the size≤3 cm),the 1,2,3 and 5-year overall survival rate was 91.7%,81.2%,60.5% and 40.4% respectively.The Cox regression models indicated that the independent factors in determining the prognosis were: Liver function of Child-Pugh stage,tumor size and occlusion of hepatic inflow during RFA.Conclusions RFA is an effective mini-invasive treatment for patients with PLC,and especially for the patients with small PLC.The efficency of RFA for small PLC has the same outcome as that of surgical resection of the tumor.Liver function of Child-Pugh stage,tumor size and occlusion of hepatic inflow during RFA are the independent prognostic factors.

Surgical treatment of thyroid cancer(a report of 246 cases) / 中国普通外科杂志

Objective To investigate the diagnosis, surgical treatment of thyroid cancer and prevention of the surgical complications. Methods Clinical data of 246 cases of thyroid cancer were analysed retrospectively from March 1990 to April 2001. Results 135 and 80 patients were diagnosed before and during operation, and all patients were diagnosed after operation; Reoperation was performed in 57 patients and residual carcinoma tissue was found in 30 patients; 1, 3 and 5 years surviving rate of thyroid cancer after operation were 99.2% , 96.3% and 91.5% respectively; No death occurred in the 246 cases after operation, 4 cases of whom showed temporary vocal cord paralysis and 3 cases showed tic due to hypocalcemia. Conclusions Intraoperative frozen section is helpful for the diagnosis and the choice of operative methods; The extention of thyroidectomy and whether to perform lymph dissection vary with the pathology types and risk factors. Patients should take thyroid tablets all their lives.

Diagnosis and treatment of nodular goiter with thyroid carcinoma / 中国普通外科杂志

Objective To investigate the diagnosis and treatment of nodular goiter with thyroid carcinoma.Methods The clinical data of forty cases of nodular goiter with thyroid carcinoma admitted to our hospital from January 2005 to December 2007 were retrospectively analyzed.Results Among the 40 cases,only four cases were preoperatively diagnosed as nodular goiter with thyroid carcinoma.All cases received operation and the diagnosis were comfirmed by frozen section examination.Various modes of thyroidectomy were performed according to the pathological results,including four cases had unilateral total thyroidectomy,27 had unilateral total thyroidectomy combined with opposite subtotal thyroidectomy,one case had bilateral subtotal thyroidectomy,and eight cases of bilateral total thyroidectomy.Of all 40 cases,11 cases received unilateral cervical lymph node dissection.There were only three cases occurred convulsion or numbness,and no hoarseness occurred postoperatively.Compared to simple nodular goiter,the incidence of calcification in nodular goiter with thyroid carcinoma was significantly increased(P