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Objective To construct the expression vectors of pSilencer3.1-hif-1? and identify the inhibition of hif-1? in Hela299 cells after transfection with the combinative plasmids.Methods The interfering sequences of hif-1? were designed according to the sequence of hif-1? of GenBank.Three recombinant plasmids of pSilencer3.1-hif-1? were constructed by DNA ligase.Trizol reagent was used to extract the whole RNA of cells and RT-PCR assay was applied to detect the expression of hif-1? mRNA.Lysis assay was used to extract the protein from cells and Western blotting was adopted to observe the expression of HIF-1? protein.Results The vectors were identified to be right after sequencing.The mRNA level was decreased 24 h after transfection with three vectors of pSilencer-hif-1?,and the ratios of hif-1? /GAPDH in control,group transfecting with pSilencer-sihif-1?-1,group transfecting with pSilencer-sihif-1?-2,group transfecting with pSilencer-sihif-1?-3 were 0.55,0.13,0.33,and 0.08,respectively(P
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Objective To study the inhibitory effects of gene combined radiotherapy on mice transplanted with B16 melanoma.Methods Alkaline lysis assay was used to extract and purify the plasmid.Plasmid DNA was injected into tumor by microinjection assay.Mice were inoculated with 5?10~5 of B16 melanoma in right hind legs and the therapy was performed when the diameter of tumor reached at 0.30.5 cm.pNE-mIL-12 and pcDNA-B7-1 plasmids were injected locally three times following with irradiation three times.The tumor growth rate of mice was observed.Results The anti-tumor effect of pNE-mIL-12 combined with pcDNA-B7-1 plasmid following with 2 Gy X-ray irradiation was much better than other groups.It showed that the tumor growth rate was slowed,the survival days of mice were delayed significantly(P
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Objective: To explore effect of TfR on immune function after ionizing by investigating changes in TfR expression on splenic lymphocytes of mice after whole body irradiation (WBI) with different dose X-rays. Methods: Direct immunoflurescence antibodies and flow cytometry were used to examine the changes of TfR expression. Results: The cell number of TfR positive expression in spleens increased significantly at 24 hours and 72 hours after WBI with 75 mGy X-rays but the cell number of TfR positive expression in spleens decreased significantly at 24 hours after WBI with 1～6 Gy X-rays. The activity of IL-2, meanwhile, demonstrated a parallel change. Conclusion: These results suggest that the TfR enhances immune function in low dose ionizing radiation but suppresses immune function in high dose. The change of TfR expression may be due to the change of IL-2 activity caused by ionizing radiation.
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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.
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Objective:To study effects of large dose thymopeptides on T cell subpopulations of patients with malignant tumor under radiotherapy.Methods:Fifty one patients with malignant tumor under radiotherapy were divided into 2 groups with 100 mg and 200 mg thymopeptides respectively.The patients we re given thymopeptides,100 mg/d or 200 mg/d,iv, for 10 days.The positive percent ages of CD4,CD8,CD25 and CD56 in T cells of peripheral blood before and after thymopeptide treatment were determined by flow cytometry.Results:The positive percentages of CD4 and CD25 in T cells of peripheral bl ood after 100 mg/d thymopeptide treatment were significantly higher than those befor e thymopeptide treatment (P<0.05),while those of CD4，CD8，CD25 and CD56 in T cells of peripheral blood after 200 mg/d thymopeptide treatment all increased significantly (P<0.05 or P<0.01). Conclusion:These results suggest that large dose of thymopeptides can increase i mmune function of patients with malignant tumor under radiotherapy,and the curat ive effect of 200 mg/d thymopeptides is better.