Objective: To study the expression and activity of the apoptosis protease-Caspase 3 in(E.coli) BL21(DE3). Methods: The cDNA of Caspase 3 was amplified by PCR and inserted into the plasmid pCMV-Myc,it is then cloned to prokaryotic expression vector pGEX-6p,after which Caspase 3 was induced by IPTG.The protein induced was identified by SDS-PAGE and Western blot. Results: After induced by IPTG for 3 hours,the concentration of Caspase 3 reached the highest level. Conclusion: Active Caspase 3 can be induced within E.coli BL21(DE3),further research can be done about the role of Caspase 3 in apoptosis.

A nucleotide diversity in DNA sequence is called polymorphism,which results in intersubjects variation in therapeutic drug effects and toxicity.Clinical studies were found that a single nucleotide polymorphism influenced response or drug toxicity in cancer chemotherapies,which were based on 5-fluorouracil,Irinotecan,platinum and so on.This review will focus on the recently development in the relation between drug effects or toxicity and polymorphism.

Objective:To investigate the effects of tetrandrine(Tet) on proliferaton and apoptosis of nasopharyngeal carcinoma cell line CNE.Methods:CNE cells were divided into groups and treated by Tet at different concentrations.Inhibitory effects of tetrandrine were determined by MTT assay.The morphologic changes of CNE cells were observed by transmission electron microscope.DNA fragmentations were determined by gel electrophoresis assay.Cell apoptosis was investigated by flow cytometer.The expressions of apoptosis-related genes were detected by retrotranscriptase polymerase chain reaction(RT-PCR).Results:Tetrandrine possessed inhibitory effect on CNE cell proliferation in a concentration-dependent manner(P

Stereotactic body radiation therapy (SBRT) adopts different tumor-killing mechanisms from the conventional fractionated radiotherapy.In SBRT,a single high-dose radiation can destroy the membrane of tumor cells and induce the release of tumor-associated antigen,also named in situ tumor vaccine,which stimulates the immune system to kill the residual tumor;a single-fraction radiation with a dose larger than 8-10 Gy can induce rapid apoptosis of vascular endothelial cells via the acid sphingomyelinase pathway at 1-6 hours after radiation,which causes tumor vascular occlusion and secondary tumor-killing effects.In order to understand whether SBRT improves the clinical treatment outcomes via the above mechanisms,this paper reviews the clinical research advances in SBRT for primary liver cancer.

Objective:To investigate the anti-tumor effect of tetrandrine on human liver cancer cell line 7402 in vitro.Effects of tetrandrine on proliferation and apoptosis of human liver cancer cell 7402 were observed.Methods:The effects of tetrandrine on proliferation of 7402 cells was observed by MTT assay and clonogenic assay.Apoptosis was observed by acridine orange(AO)/ethidium bromide(EB) Fluorescent staining?DNA gel electrophoresis and flow cytometry,and the expression of apoptosis related gene was analyzed by immunohistochemical staining method.Results:Tetrandrine inhibits the proliferation of 7402 cells in a dose dependent manner and induces apoptosis.Immunohistochemical staining showed that the expression of Bcl-2 was decreased and the expression of Bax was increasd in 7402 cells treated with tetrandrine.Conclusion:Tetrandrine inhibits the proliferation of 7402 in the dose dependent manner,and induces apoptosis.The antitumor effect of tetrandrine may be due to the regulation of the expressions of Bcl-2 and Bax.

Human gliomas are one of the most aggressive tumors in brain.Radiotherapy plays an important role for patients with gliomas,as well as surgery.The efficacy of radiotherapy is associated with radiosensitivity of human gliomas.Radiosensitive genes of gliomas and apoptosis、cell cycle transformation、DNA damage induced by irradiation and DNA repair promoted by them are suggested to be associated with glioma radiosensitivity.The rediosensitive genes associated with apoptosis and DNA repairment are becoming hot spots of study.The other radiosensitivity genes are paid attention too.This paper makes a summary of current situation and progress for radiosensitive genes of human brain glomas.

Objective: To compare the difference of intensity modulated radiation therapy (IMRT),3-D imensional conformal radiation therapy (3DCRT) for patients with upper esophageal carcinoma. Methods: Ten patients with upper esophageal carcinoma were treated by intensity modulated radiation therapy and 3-D imensional conformal radiation therapy at the same TPS, the difference of exposure dose between target area and critical organ was compared by dose volume histogram(DVH) with the plan target volume (PTV) must reach 95% of the prescription dose. Results: There was significant difference in dose of 95% plan target volume (PTV) (P <0.05) IMRT better than 3D-CRT. For two target conformal index and the prescription dose coverage of GTV percentage IMRT was better than 3DCRT. IMRT reduced maximum dose of spinal cord (P <0.05). There was no difference in the dose of lung and heart (P >0.05). Compared with 3D-CRT, IMRT planning has better dose distribution and protection of normal tissue. Conclusions: IMRT was better than 3DCRT, IMRT is the best radiation therapy.

Objective: To construct and identify a lentiviral vector harboring RNAi sequence targeting the human high mobility group A1(HMGA1) gene.Methods: The effective sequence of siRNA targeting the HMGA1 gene confirmed in our previous study,the complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCL-GFP vector diced by the restriction enzyme of HpaⅠ and XhoⅠ,which contained the U6 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing HMGA1 shRNA was named LV-sh HMGA1 and confirmed by PCR and DNA sequencing.A total of 293T cells were cotransfected with LV-sh HMGA1,pHelper 1.0 and pHelper 2.0.All the virus stocks were produced by Lipofectamine2000-mediated transfection.The titer of the virus was tested according to the expression level of GFP.Results: PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the human HMGA1 gene was successfully inserted into the lentiviral vector.The titer of the recombinant lentiviral vector was 5?107 TU/ml.Conclusion: The successful construction of the lentiviral vector of HMGA1 has prepared the ground for further studies on the functions of the HMGA1 gene with the RNAi technique.

Objective To retrospectively analyze the engraftment, transplant-related complications and survival after unrelated cord blood transplantation (UCBT) in patients with hematologic malignancies. Methods Fifty consecutive patients with hematological malignancies (median age, 19 years; median weight, 53 kg) were treated with UCBT in single center from April 2000 to August 2009. Thirty-nine patients were high-risk or refractory. Double UCB grafts were used for 26 patients, while single UCB graft for 24 patients. Myeloablative conditioning was given to 45 cases and non-myeloablative regimens to 5 cases. All patients were given a combination of cyclosporin A (CsA) and mycophenolate mofetil (MMF) for graft-versus-host disease (GVHD) prophylaxis. Results The median total nucleated cell (TNC) dose was 4.0 (range, 1.95-16.24)×10~7 TNC/ kginfused, and CD34~+ cell dose was 2.74(range, 0.67-29.28)×10~5/kginfused. Forty-two of 50 patients acquired engraftment with implantation rate being 86%. The median time to engraftment (absolute neutrophil count>500/mm~3 and platelets 20 000/L) was 19 and 34 days. The cumulative incidence of neutrophil engraftment by day 42 was 86.3%(95% confidence interval [CI] 0.769-0.957); the cumulative incidence of platelets engraftment by day 120 was 72.3% (95% CI 0.620-0.821). Twenty cases developed acute GVHD, and the incidence of acute GVHD of grades Ⅲ/Ⅳ by day 100 was 7.1%. The incidence of chronic GVHD within 2 years was 17.4%. During a median follow-up period of 22 months (range 4-116), Overall 6-month, 1-year and 2-year survival rate was 66.2%(95% CI 0.590-0.734), 57.4%(95% CI 0.496-0.652), 54.2%(95% CI 0.462-0.622), respectively. For the patients with non-advanced hemotologic malignancies, 6-month, 1-year and 2-year survival rate was 73.2% (95% CI 0.659-0.805), 66.1% (95% CI 0.579-0.743), and 62.2% (95% CI 0.542-0.682) respectively. Five cases relapsed. The cumulative incidence of relapse within 2 years was 16.2% (95% CI 0.099-0.225). Twenty-one cases died mainly due to infection. Conclusion UCBT could be safely and effectively used for adult patients with hematologic malignancies.

Objective To investigate the radiosensitiation effect of berberine on human nasopharyngeal carcinoma ( NPC) in hypoxia condition and explore the underlying mechanisms. Methods MTT assay, clonogenic assay and flow cytometry were performed to analyze cell proliferation, colony formation and apoptosis, respectively. Male nude mice inoculated subcutaneously with CNE-2 cells were used to examine the radiosensitization effect of berberine in vivo. The expressions of HIF-1α and VEGF were assessed by Western blot. Results Berberine efficiently inhibited the proliferation of CNE-2 cells in time-dependent and dose-dependent fashions with an IC50 of ( 14?9 ± 2?2 ) μmol/L. Clonogenic survival assay showed that berberine ( 5 μmol/L ) sensitized CNE-2 cells to ionizing radiation in hypoxia and its SERD0 was 1?27. Under hypoxic condition, berberine alone (5, 15 μmol/L) could induce apoptosis (t=5?01, 9?02,P<0?05) and it further promoted 8 Gy radiation-induced apoptosis (t =5?31, 9?91,P <0?05). Moreover, berberine significantly delayed the tumor growth in the combination group (berberine +irradiation) compared with the mice received irradiation alone or PBS (t =2?96, 14?52, P <0?05). Immunobloting assay showed that berberine inhibited the upregulation of HIF-1α and VEGF induced by hypoxia in CNE-2 cells. Conclusion Berberine confers radiosensitivity on hypoxic NPC in vitro and in vivo, which is probably associated with the downregulation of HIF-1α and VEGF expressions.