Pim-3,a new member of the the proto-oncogene pim family which expresses serine and threonine kinase activity,shows a high similar to pim-1 and pim-2 at the function,structure and sequence.Pim-3 plays an important regulative role in the cell cycle transform,cell apoptosis and occurrence of the malignant tumors by phosphorylating numerous specific substrates.

10.3969/j.issn.2095-4344.2013.24.004

Objective: To investigate the anti-tumor efficiency of B16 melanoma HACs in vivo and in vitro.Methods: Tris-HCI extract and Sephacryl S-200 gel filtration were applied to prepare B16 HACs, and the cytolokicity of specific CTL induced by HACs was tested. Results: The 41, 47 and 53 tube HACs obtained by gel filtration could decrease the tumor incidences, delay the time of tumor development and decrease the mortalitites of mice.Conclusion:60 ~ 97 kD HACs from B16 melanoma cytosol have the activites of inhibiting tumor and could be used in effective anti-tumor therapy.

Objective: In the present study we observed the general pattern of the adaptive response of thymocyte apoptosis and cell cycle progression induced by low dose radiation (LDR). Methods: Kunming male mice were irradiated with the inductive dose (D1, 75 mGy) and the challenging dose (D2, 1.5 Gy). The intervals between D1 and D2 were 3, 6, 12, 24 and 60 hours. The changes of thymocyte apoptotic bodies (TAB) and cell cycle progression were measured with flow cytometry with the thymocytes cultured for 4, 20 and 44 hours, respectively, 18 hours after irradiation with D2. Results: When the intervals between D1 and D2 were 3, 6 and 12 hours, the percentages of TAB in the D1 + D2 groups in the thymocytes cultured for 4 and 20 hours were significantly lower than those in the D2 groups (P<0.05) and the percentages of G0/G1 and G2 + M phase cells decreased in varying degrees, while the percentages of S phase cells increased significantly (P<0.05 or P<0.01). Conclusion: The results mentioned above indicate that when the mice were irradiated with 75 mGy (D1, 12.5 mGy/min) 3～12 hours before 1.5 Gy (D2, 0.285 Gy/min) exposure, the adaptive response of apoptosis and cell cycle progression may be induced with the thymocytes cultured for 4 and 20 hours after whole-body irradiation with D2.

Objective To construct the conditionally replicative adenovirus vector pAd-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55K carrying early growth response gene-1 (Egr1)promoter and tumor necrosis factor related apoptosis inducing ligand (TRAIL)gene, and to observe the effects of the vector combined with 2 Gy irradiation on the TRAIL expression in MDA-MB-231 cells.Methods Egr-1 promotor sequence was cloned from pMD18 T-Egr1, TRAIL was constructed the downstream of Egr1 promoter, pShuttle-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55K (CRAd.pEgr1-TRAIL)was constructed,after the adenovirus vector was packaged successfully,MDA-MB-231 cells were infected with them and irradiated with X-rays.Real time PCR method and ELISA were used to detect the expression levels of TRAIL mRNA and protein, respectively. Six groups in the experiment were set up:control, 2 Gy,CRAd.p,CRAd.pEgr1-TRAIL,CRAd.p + 2 Gy and CRAd.pEgr1-TRAIL + 2 Gy. Results The recombinant adenovirus vector pAd-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55K was constructed and packaged successfully.The expression level of TRAIL mRNA in MDA-MB-231 cells transfected with the vector of 5 MOI for 24 h following 2.0 Gy X-rays irradiation began to increase and arrived to the top 8 h later in various groups,then declined.The expression level of TRAIL protein in MDA-MB-231 cells began to increase 6 h after irradiation and reached to the peak 24 h later,then declined 48 h later.There were significant differences in the expression levels of TRAIL protein between CRAd.pEgr1-TRAIL + 2.0 Gy and other groups at the same time point (P<0.01). Conclusion The recombinant adenovirus vector is obtained successfully, and the TRAIL mRNA and protein expression levels in MDA-MB-231 cells can be increased significantly by the vector combined with 2.0 Gy X-rays irradiation.

This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellular growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshuttle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G(2)/M phase and apoptotic rate was increased, and the percentage of cells at G(0)/G(1) phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.

This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellular growth,apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro.The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egrl-shTRAIL-shES and X-ray irradiation.Then MTT assay was used for determining the cellular proliferation,and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression.The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshuttle-Egrl-shTRAIL-shES transfection in conjunction with irradiation.In the TRAIL-endostatin-based single- or double-gene-radiotherapy,the cell viability declined in a time- and dose-dependent manner,the percentage of cells at G2/M phase and apoptotic rate was increased,and the percentage of cells at G0/G1 phase was lowered as compared with those receiving radiotherapy alone.Moreover,TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition,promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.

Objective: To investigate IMRT planning process using the combined application of failure modes and effects analysis (FMEA) and fault tree analysis (FTA) by reference to the report of Task Group 100 of the AAPM, and stablish and optimize the quality. Methods: A multidisciplinary team detailed the process mapping of IMRT planning using Eclipse TPS. The team evaluated the potential failure modes (FMs) of every process step. The evaluation was divided into two groups according to whether quality management (QM) was considered. For every FM, occurrence (O), severity (S) and detectability (D) by consensus were evaluated, and the product of O, S and D yielded the risk priority number (RPN), which permitted the ranking of the FMs. Finally, FTA was used to determine the root factors contributing to the riskiest failure modes. Results: The IMRT plan process consisted of 10 major sub-processes and 33 steps, which amounted to 47 failure modes. For the group without quality management, the RPN of FMs was between 13.2-271.8, 27 of which had RPN≥80, and 18 FMs had S≥8. For the group with quality management, the RPN of FMs was between 11.2-158.4, 11 of which had RPN≥80. The difference of RPN between the two groups was statistically significant (RPN of the group without QM=101.17±66.34, RPN of the group with QM=59.54±35.64, t=8.501, P<0.05). Finally, FTA was used to determine the root factors contributing to the FMs, i. e., prescription dose definition and importing images. Conclusions The FMEA and FTA methods are operable and practical, which can systematically and comprehensively analyze the potential failures and risks existing in the process of IMRT plan. And the FMEA and FTA can contribute to establish and optimize the quality management program in radiotherapy.