The in vivo tumorigenicity of murine B16 melanoma cells engineered to secret TNF-a was observed. The retrovirus containing mouse TNF-a cDNA was generated by the virus-packing cell PA317 transfected with plasmid pXT-TNF. The B16 cell clone secreting the highest TNF-a level was obtained after G418 resistance selection, limiting dilution and the assay of TNF-a activity. After the mice were inoculated subcutaneously with the cell clone, we found the tumor growth was inhibited and the survival period of the mice extended when compared with the mice inoculated with the wild-type B16 cells . We also found that the tuinorigenicity of B16-TNF-a+ cell was associated with the cell number inoculated. At or above the 1.25? 104 cells, the percentage of the mice with detectable tumor correlated negatively with the cell number inoculated: however, at the 6.25 ? 103 cells, the percentage was higher than that at 2.5?10~(4) cells. These results encourage us to do further experiments on the following tumor cell-targeted TNF-a gene therapy.

In the present study, the effects of G-CSF gene therapy was investigated on the recovery of murine hematopoietic suppression induced by high dose of cyclophosphomide (Cy) . The results showed that G-CSF gene therapy could slow down the Cy-induced decreasing of peripheral WBCs and platelets, and accerelate their recovery.It also could increase the CFU-GM, CFU - MK, CFU-S derived from the splenocyte and bone marrow cells in the chemotherapy-treated mice. The data demonstrated that fibroblast-mediated G-CSF gene therapy could significantlyreduce the hematopoietic damage to less extent, and accerelate hematopoietic recovery after chemotherapy.

Objective To explore the chemotherapeutic agents-induced modulating effects of Egr-1 promoter sequence on GM-CSF expression and its protective effect against injury to haematopoiesis due to chemotherapy.Methods Human GM-CSF cDNA and enhanced green fluorescent protein(EGFP) cDNA were linked to internal ribosome entry site(IRES) respectively,and then recombined to pCIneo vector containing Egr-1 promoter(Egr-EG).The vector was transferred into human bone marrow stromal cell line HFCL by lipofection,and the HFCL/EG cells were then finally constructed.MTT assay was performed to determine the effects of cisplatin(DDP),5-fluorouracil(5-FU),gemcitabine(GEM) and paclitaxel(PTX) on the survival rates of HFCL/EG and HFCL cells,and the IC50 of such agents against HFCL/EG and HFCL cells were calculated.The percentage of HFCL/EG cells which positively expressed EGFP was assessed by both flow cytometry and inverted fluorescence microscope.The effects of the active oxygen inhibitor N-acetylcysteine(NAC) on the expression of GM-CSF,which was modulated by promotor Egr-1,were detected by ELISA.The effects of HFCL/EG supernatant on CFU-GM after being exposed to chemotherapeutic agents were examined.Results With different sensitivity to DDP,5-FU,GEM and PTX,HFCL/EG cells were successfully constructed.The drugs showed higher IC50 value against HFCL/EG cells than HFCL cells(P

Radiotherapy is one of major cancer treatment methods.However,radiation resistance is an important reason to restrict the efficacy of radiotherapy and lead to treatment failure.In recent years,the relationship between the canonical Wnt signaling pathway and tumor radiation resistance has more and more attention of the scholars.This review summarized recent ten years findings concerning the canonical Wnt signaling pathway and tumor radiation resistance and tried to find some valuable rules or some internal relationships among different pathways by systemically analyzing.

Objective To investigate the effects of improved end-to-end invagination pancreaticojejunostomy on the occurrence of pancreatic fistula after pancreaticoduodenectomy.Methods The clinical data of 396 patients who received pancreaticoduodenectomy at the Cancer Hospital of Shandong Province from January 2001 to January 2011 were retrospectively analyzed.All patients were divided into the improved group(235 patients)and traditional group(161 patients)according to different anastomotic methods.All the operations were done by the same surgical group,and the digestive tract was reconstructed by the Child method.Patients in the improved group received improved end-to-end invagination pancreaticojejunostomy,and patients in the traditional group received traditional end-to-end anastomosis.The volume of operative bleeding,operation time,incidence of pancreatic fistula and duration of hospital stay of the 2 groups were compared.All data were analyzed using the t test,chisquare test or Fisher exact probability.Results The operative blood loss,operation time and duration of hospital stay were(383 ±56)ml,(7.2 ± 1.0)hours,(21 ±3)days in the improved group,and(381 ±39)ml,(7.0 ± 0.5)hours,(22 ± 5)days in the traditional group,with no significant difference between the 2 groups(t =0.388,1.680,-1.835,P ＞ 0.05).No operative death was detected in the 2 groups,and the overall incidence of pancreatic fistula was 7.6％(30/396).The incidence of pancreatic fistula of the improved group was 0(0/235),which was significantly lower than 18.6％(30/161)of the traditional group(P ＜ 0.05).Patients complicated with pancreatic fistula in the traditional group were cured by drainage,somatostatin administration and parenteral nutrition.Conclusion Improved end-to-end invagination pancreaticojejunostomy can significantly reduce the incidence of pancreatic fistula after pancreaticoduodenectomy.

Objective To evaluate complications from two anal stapling operations of anorectal surgery,the procedure for prolapse and hemorrhoids (PPH) and the stapled transanal rectal resection (STARR).Methods The data of 1 276 patients undergoing PPH for hemorrhoids and 149 patients having STARR for ODS from January 2010 to January 2015,in Beijing Chaoyang Hospital were studied.More than six months of follow-up was done.Statistics used included t Test and Chi-square test,Logistic regression analysis.Results Complications of PPH and STARR included hemorrhage in 27 cases (2.1％) vs.3 cases (2.0％);pain in 285 cases (22.3％) vs.6 cases (4.0％);uroschesis in 96 cases (7.5％) vs.12 cases (8.1％);edema in 227 cases (17.8％) vs.16 cases (10.7％);defecatory urgency in 194 cases (15.2％) vs.38 cases (25.5％);anastomotic infection in 17 cases (1.3％) vs.2 cases (1.3％);mild anal incontinence in 11 cases (0.9％) vs.2 cases (1.3％);anastomotic stenosis in 11 cases (0.9％) vs.1 case (0.7％).There was a positive correlation between dry stool,defecatory urgency and hemorrhage after PPH.PPH compared to STARR:The post-operative pain,edema and defecatory urgency and overall complication rate was significantly different (t =26.51,x2 =4.69,x2 =10.38,x2 =37.12,P ＜ 0.05).Conclusions PPH and STARR have rare serious complications and easy to handle.Abnormal defecation such as dry stool and defecatory urgency is an important risk of hemorrhage.

Objective To study the results of laparotomy and Cool-tip radiofrequency ablation to treat large liver tumors.Methods Laparotomy and Cool-tip radiofrequency ablation were carried out on 64 patients with large hepatic cancer.To destroy the tumor completely,for tumors of 3.0～4.0 cm in diameter,7 ablations were required; for 4.0～5.0 cm in diameter 15 ablations; for 5.0～6.0 cm in diameter 19 ablations; for 6.0～7.0 cm in diameter 40 ablations.Result The complete necrosis rate of laparotomy and radiofrequency ablation was 93.75％ (60/64).The short-term results were good.Conclusions Laparotomy and Cool-tip multipoint overlapping radiofrequency ablation for large liver tumors (tumor diameter＞3 cm) could result in a high complete necrosis rate and a low complication rate.It is a good radical treatment for unresectable and large liver cancer.

Objective: To compare the pharmacokinetics and bioavailability of verapamil hydrochloride pulsed release tablets with core tablets. Methods: Latin test was employed in the single oral administration of the Ⅲ,Ⅳ type of pulsed release tablets and core tablets in 8 volunteers. The pharmaceutics behavior of the tablet in vivo was evaluated by the lag time, c max ,AUC and so on. Results: The pharmacokinetics results demonstrated that the Ⅲ type of pulsed tablet in humans could be released after about 4 h lag time. In a proper range, pulsed release tablets only changed the beginning time while c max and AUC were not different from the core tablets. Conclusion: A new system to reduce the early morning symptoms of ischemic heart disease is prepared. [

Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+,　CD4+,　CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.