Telomerase, as a factor of tumour's happening and development, is a kind of cell element not existing or with low activity in normal cells. Moreover, the activity of telomerase is a common channel for cell's immortalization and deterioration, and is also the main factor in cloning tumour and developing malignant turnour. Therefore, the tumour gene therapy focusing on telomerase has become the new target of present study of tumor.This therapy has greater prospects on clinical application than the past tumour gene therapy simply directing at one cancer gene.

As a T cell subsets growth factor,interleukin-2 (IL-2) can inhibit cancer cell proliferation,reduce tumorigenicity and transitivity,improve local tumor microenvironment,enhance the immunogenicity of hepatocellular carcinoma cells,reduce the load of hepatitis virus and greatly enhance the ability of anti-tumor.On the other hand,it can enhance tumor hypercoagulable state.The effect of IL-2 on primary hepatic carcinoma and its potential clinical value have a bright future.

Lentiviral vectors (LV),a special member of retroviral vectors,has become hot in gene therapy research.Studies have shown that LV can make exogenous gene expressed stably in tumor cells.As a tool for transportation,LV plays a significant role in the gene therapy of digestive system,hematologic system and gynecological tumor,which provides a promising new way for neoplasms gene therapy.

Objective To observe the effect of human telomerase reverse transcriptase-thymidine kinase/ganciclovir (phTERT-TK/GCV) system combined human telomerase reverse transcriptase-tumstatin (phTERT-tumstatin) system on apoptosis of human HepG2 and mRNA expression and protein content of AFP,RhoC related with cancer.Methods Fluorescence microscopy was used to observe expression of EGFP and MCHERRY in transfected HepG2 and L-02.Real-time PCR and Western blot were used to detect AFP and RhoC mRNA and protein content.Flow cytometry was used to detect the apoptosis of transfected HepG2.Results phTERT-tumstatin and phTERT-TK/GCV genes expressed in transfected HepG2.Real-time PCR showed that AFP and RhoC mRNA expression in different group were 0.76±0.09 and 0.80±0.04 (TK/GCV group),0.62±0.09 and 0.40±0.02 (TM group),0.49±0.07 and 0.54±0.03 (MK group).The differences were significant (P ＜ 0.01) except TK/GCV group compared with empty plasmid group.Western blot test results showed that protein content of AFP and RhoC were higher in TK/GCV group (0.97±0.02/1.17± 0.01),TM group (0.83±0.02/0.99±0.02),MK group (0.69±0.01/0.77±0.02) than in empty plasmid group (1.19±0.03/1.32±0.05) and non-transfected group (1.15±0.05/1.29±0.30) (P ＜ 0.01).Additionally,protein content of AFP and RhoC in MK group were significant difference with TK/GCV group and TM group (P ＜ 0.01).Flow cytometry showed that phTERT-tumstatin,phTERT-TK/GCV co-transfected HepG2 cells apoptosis rate was significantly higher than both individually transected group.Cells apoptosis rate of alone and co-transfected groups was significant difference compared with empty vector group and untransfected group.Conclusions The effect of phTERT-TK/GCV and phTERT-tumstatin on pro-apoptotic of HepG2 cells is significant.phTERT-TK/GCV combined with phTERT-tumstatin has strong therapeutic function.

Objective To observe if IL-2 gene can express in hepatic carcinoma cells by double targeting of hepatitis B virus envelope (HBVE) and alpha fetoprotein (AFP) promoter.Methods HepG2 cells,L02 cells,and HepG2.2.15 cells were cultured in vitro.HBVE was obtained by PEG8000 concentration assay,and the acquired HBVE was used to pack recombintional gene.Human AFP promoter-IL-2 recombinational gene was obtained by PCR.Then HepG2 cells and L02 cells were transfected by transient transfection and the expression of IL-2 was detected by RT-PCR and Western blot.Results IL-2 was detected in HepG2 cells by RT-PCR and Western blot but not in L02 cells.Conclusion By using HBVE as a gene transporter,human AFP promoter-IL-2 recombinational gene can express in hepatic carcinoma cells,thereby it can increase the safety of exogenous gene transfection of hepatic carcinoma cells.

The process of apoptosis runs through a variety of signal transduction, and is regulated by many apoptosis-related genes. Inhibitors of apoptosis protein (IAPs) are closely related to the infinite proliferation of tumor cells. IAPs including the recently discovered survivin, livin, are closely related to tumors. The anti-apoptotic mechanism is to be further studied.

The hTERT gene promoter is a tumor-specific promoter, regulated by a number of factors.Modulation of the chromatin structure via DNA methylation or histone acetylation plays an important role in regulating the hTERT promoter. Recent study also found that glycogen synthase kinase -3 cytokines, macrophage colony-stimulating factor receptor 1, bone inducing factor -7, transcription factors c-myc inhibitor, transforming growth factor β, cyclopentenone prostaglandin regulate the activity of hTERT through various pathways. Telomelysin, the telomerase-specific replication competent oncolytic adenovirus, has been used in clinical settings.Various gene therapies based on hTERT are current under-way.

Objective: To observe the effect of ultrasound-guided clavicular brachial plexus block in upper limb surgery.Methods: Eighty patients undergoing upper limb surgery were enrolled and randomly divided into two groups: ultrasound-guided clavicular brachial plexus block group (ultrasound guidance group, n=40) and nerve stimulator-assisted positioning of the supraclavicular brachial plexus block group (nerve stimulator group,n =40).The block effect, anesthesia effect, anesthesia completion time, nerve block onset time, nerve block duration and complication were compared and analyzed statistically between the groups.Results: The completed rate of block was 97.5%in the ultrasound guidance group, which was significantly higher than that in the nerve stimulator group (65.0%) (P<0.05);the uncompleted rate was significantly lower than that in the nerve stimulator group.The fine/excellent rate of anesthesia was 95.0% , which was significantly higher than that of the nerve stimulator group (75.0%, 30/40) (P<0.05);the complete time of anesthesia and nerve block onset time were significantly shorter than those in the nerve stimulator group (P<0.05);the duration of nerve block was significantly longer than that in the nerve stimulator group (P<0.05);the incidence of complications was 7.5%), which was significantly lower than that of the nerve stimulator group (37.5%, 15/40) (P<0.05).Conclusion: In upper limb surgery, ultrasound-guided nerve stimulator assisted positioning of clavicular brachial plexus block is better than nerve stimulator assisted positioning of clavicular brachial plexus block.

Objective To compare the transfection efficiency of galactosylated chitosan nanoparticle vehicle with chitosan nanoparticles vehicle,and observe the therapeutic effect of pGL3-hTERTp-TK on HCC cell line HepG2. Methods Preparing the chitosan and galaetosylated chitosan. Constructing the pGL3-hTERTp-TK plasmid and the Ch/DNA and GC/DNA complexes.Transfecting the HepG2 and the normal hepatic cell L-02 with chitosan/DNA and galactosylated chitosan/DNA complexs.Detecting the fluorescence and the expression of luciferase gene using the fluorescent microscope and the scintillation counter.Detecting the cell growth and apoptosis through the Caspase-3 and the flow cytometry. Results Two clear straps appeared in the agarose gel.The locations were 300 bp and 1100 bp.The relative lueiferase activity of pGL3-hTERTp-Luc+mediated by galactosylated chitosan was powerful than which of pGL3-hTERTp-Luc+mediated by chitosan in HepG2 by the scintillation counter.However,the relative luciferase activity was very weak in L-02.The same results were observed by fluorescent microscope.When the concertration of the GCV was 10 μg/ml(t=51.40,P=0.000),the HepG2 cell inhibition which was transfected by GC-pGL3-hTERTp-TK was obviously different from the L-02 cell inhibition which was transfected by GC-pGL3-hTERTp-TK in the statistics.The significant apoptotic rate was 65.28%in HepG2 which was transfected with GC/DNA,whereas it was only 10.80% in L-02. The significant apoptotic rate in HepG2 which was transfected with GC/DNA was very higher than the other groups (LSD, P ＜0.05). The significant apoptotic rate (10.80%) in L-02 which was transfected with GC/DNA was very higher than the group which was Ch-pGL3-control (LSD, P =0.000). The average fluorescence intensity of the HepG2 which is transfected by the Ch-pGL3-hTERTp-TK was 168.02± 3.68. The average fluorescence intensity of the HepG2 which is transfected by the GC-pGL3-hTERTp-TK was 204.45 ±3.45. The two groups had a significant difference in the statistics (t=-12.504, P＜0.05). Conclusion Galactosylated chitosan has higher transfection efficiency than chitosan. GC-pGL3-hTERTp-TK could specially attack HCC cell line and almost has no influence on normal hepatic cells.

Objective: To study the effect of intracellular-free calcium and the expression of Fas and Fas L on the process of pancreatic carcinoma cell apoptosis. Methods: Apoptosis induced by 2 μmol/L arsenic trioxide in pancreatic cancer cell lines SW-1990 was investigated.Concentration of intracellular-free calcium （［Ca2+］i） was determined by Fura-2a fluorescein load technique. Fas and FasL were determined by flow cytometry. Results: Pancreatic cancer cells treated with 2 μmol/L arsenic trioxide presented apoptotic features: intact cell membrane, chromatin condensation, nucleic fragmentation and apoptotic body formation; agarose electrophoresis showed marked ladder; flow cytometery analysis showed a sub-G1 cell peak. In the process of pancreatic carcinoma cell apoptosis Fas and FasL and the ［Ca2+］i were significantly higher than that in the control. Conclusion: The pancreatic cancer cell apoptosis induced by arsenic trioxide is related to Fas and FasL expression by the cancer cells and the ［Ca2+］i increase in the cancer cells.