We used retroviral vector pLXSN to construct recombinant retroviral vectors with the human apoptosis gene, interleukin-l? converting enzyme (ICE). The vectors were introduced into packaging cell line PA317 by electroporation method. The G418 resistant colonies were selected, and the supematants of the colonies were used to infect the human hepatocellular carcinoma cell line SMMC7721. G418 resistant colonies of SMMC7721 were named SMMC7721-MCE and SMMC7721-neo. The results of RT-PCR analysis showed that exogenous hICE gene had successfully integrated into the genome of SMMC7721-hICE cells. The proliferation rate and tumorigenicity of cells in nude mice were examined. Our data showed that the growth rate and the tumorigenicity of SMMC7721-hICE cells in nude mice were considerablely decreased comparing with parent SMMC7721 and SMMC7721-neo. These results suggested that the retroviral vector expressing hICE gene was successfully constructed and could suppress the growth ability and tumorigenicity of human hepatocellular carcinoma cells, which provided a basis for further investigation of hICE gene therapy.

Cytotoxic T lymphowcytes (CTL) play a major role in host anti-tumor immune responses. A major obstacle to the application of adoptive immunotherapy in the treatment of human maligancy is the inability to generate enough activated CTLs since the cytotoxic T cell undergoes activation induced apoptosis during destroying tumor cells. It is important to study how to limit activaton induced apoptosis of T cell so as to maximize the number of CTL and to enhance the tumor cytotoxicity. We have used CD3-induced Jurkat cell line as an activated T cell apoptosis model and introduced the anti-sense ICE cDNA into Jurkat T cells with retroviral vector. The effect on apoptosis of Jurkat cell induced by anti-CD3 or anti-Fas monoclonal antibody was evaluated after transfection with antisense human ICE. We found that the level of ICE expression in Jurkat cell transduced by the vector decreased and apoptosis was minimized in antisense ICE-transfected Jurkat cell after anti-CD3 or anti-Fas treatment. These results suggest that antisense blocking of ICE expression can partially inhibit Jurkat cell apoptosis induced by anti-CD3 or anti-Fas. Thus, applying antisense blocking of ICE to gene therapy may block the apoptosis of activated T cells, furthennore, enhance the antitumor effect.

Objective To evaluate the efficacy of laparoscopic-assisted resection of the small bowel for stromal tumors.Methods From January 2003 to May 2007,laparoscopic abdominal exploration was carried out under general anesthesia on 10 patients with space occupying lesion and 5 patients with abdominal pain and hemafecia of unknown origin.After an intestinal lesion was found,a small incision was made at the proper site on the abdominal wall according to the location of the lesion.The diseased intestine was resected outside the abdominal cavity,and then end-to-end anastomosis was performed.Pneumoperitoneum was rebuilt after closing the abdominal cavity in order to observe the blood circulation of the small bowel and intra-abdominal hemorrhage.Results The operation was completed in all the 15 cases.The postoperative pathological examination showed stromal tumors with CD117 positive in 14 cases(93%)and CD34 positive in 9(60%).Three(20%)of the patients were at very low risk,5(33%)at low risk,4(27%)at moderate risk,and 3(20%)at high risk.The mean diameter of the tumors was(2.44?0.63)cm(ranged from 1.5 to 3.6).The operation time was 38 to 72 minutes with a mean of(57.8?10.4),and mean blood loss was(20.1?6.5)ml(10 to 30).The patients were discharged 4 to 7 days after the operation [mean,(5.3?1.1)days],and were followed up for 5 to 36 months [mean,(24.3?8.4)months].No patient had postoperative complications or recurrence in this series.Conclusion Laparoscopy is effective for resection of small bowel stromal tumors.

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.

Cancer is one of the main diseases that threaten the health of human.In order to increase the cure rate of cancer,the accurate therapy of cancer must be developed speedily.The most effective method for curing cancer is intensity modulated radiation therapy(IMRT),which can increase the local control rate of cancer and decrease complications of tissues.IMRT is considered to be an important breakthrough in cancer therapy.The dose produced by it is better than 3D-CRT and can achieves better results of therapy,which has been confirmed in the clinical therapies of head cancer,neck cancer,prostate cancer,breast cancer,cervical cancer and pancreas cancer etc.The developing course of IMRT,the methods of intensity modulation,the enforcement process of IMRT and the feasibility test of therapy plans are emphatically introduced.

Objective To construct adenovirus vector with agiostatinK1-5 gene and to investigate the function of suppression to proliferation and migration for vascular endothelial cells.Methods With the use of gene recombination and clone technology, we constructed the adenovirus vector with the gene of angiostatin K1-5. In vitro vascular endothelial eclls proliferation assay and migration activity were performed through direct infection,MTT and transwell chemotaxis assay. Results 50% TCID indicated that the condence of resultant viruses was 1.5?109PFU/mL. It was purified by CsCL banding,final yield were generally 1.1?1010 PFU/mL plaquing-forming units. Through indirect infect assay and MTT, we found angiostatin K1-5 inhibited human vascular endothelial cells proliferation. We utilized human vascular endothelial cells to study the effect angiostatin K1-5 on cell migration,the result showed that adenoviruse vector with angiostatin K1-5 significantly inhibited HUVEC migration.Conclusion We successfully constructed adenoviruse vector with angiostatin K1-5 and demonstrated its inhibitory effect on proliferation andmigration of HUVEC.

Possessing ideological qualities is a gradual process. We should establish a multiplex, synthetical and entire assessment system for the ideological and political theory course exams in medical colleges. The innovation of exam mode can promote the diversification of teaching style.

Objective To study the protection against hepatic ischemia-reperfusion injury by human IL-10 gene transduction in rats. Methods Ad-hIL10-EGFP (1. 0 ? 109 plaque forming units/ml) was administered into SD rats by intravenous injection 72 hours before hepatic ischemia-reperfusion injury was induced. Liver function were tested and HE pathology was observed. The expression of hIL-10 was studied with ELISA or immunohistochemical method, the expression of EGFP was observed in frozen sections under the fluoroscopy. The apoptosis of hepatocytes was observed with Tunel's assay. Results Compared with control rats, the expression of EGFP and hIL-10 was observed, serum hIL-10 level was (815.74 ? 284. 76) ng/ml, liver function of treatment rats were improved, the paraffin sections showed that the hepatocytes were not significantly swelling and liver pathology ameliorated, the number of apoptosis cells decreased (P

Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

Objective To explore the effect of 1a,25(OH)2 D3 on circadian clock gene expressions in cardiac myocytes.Methods Cultured cardiac myocytes isolated from 7 -day -old Sprague -Dawley(SD)rats were identified by immunofluorescence.The medium including 1a,25 (OH)2 D3 (final concentrations were 0 nmol/L,1 nmol/L, 10 nmol/L,50 nmol/L and 100 nmol/L)were added to primary myocardial cells to culture for 2 h and then total RNA was extracted.Real -time polymerase chain reaction (RT -PCR)was applied to analyze myocardial cells circadian clock gene (Bmal1,Per2,Rev -erba)transcript levels to determine optimum concentration of 1a,25(OH)2 D3 .Then, the primary myocardial cells cultured for 72 h were divided into 3 groups:the control group was of serum -free culture medium;serum shock group was of DMEMcontaining 50%volume fraction of horse serum cultured 2 h;1a,25(OH)2 D3 treatment group receiving 1a,25 (OH)2 D3 at optimal concentration cultured 2 h.The cells were collected at 7 time points (0 h,4 h,8 h,12 h,16 h,20 h,24 h)and then total RNA was extracted.RT -PCR was applied to analyze circa-dian clock gene (Bmal1,Per2,Rev -erba)transcript levels in the myocardial cells.Results In the presence of 50 nmol/L 1 a,25(OH)2 D3 ,the Bmal1 mRNA expression showed the highest level,but the Per2 and Rev -erba mRNA expression levels were minimum.Compared with the control group,both 1a,25 (OH)2 D3 treatment group and serum shock group caused day -cycle rhythmic oscillation in circadian clock genes(Bmal1,Per2,Rev -erba)in the cardiac myocytes.And the expressions pattern of Bmal1 and Per2 genes were in the opposite phase.While Bmal1 gene expres-sion appeared at peak at 12 h,Per2 gene expression appeared in a trough.Expression of Rev -erba gene trend began to rise at 8 h,and the highest expression level appeared at 12 -16 h.Conclusions 1a,25(OH)2 D3 can affect Bmal1, Per2 and Rev -erba mRNA expressions of circadian clock genes in the cardiac myocytes.