Application of Flt-3 Ligand in serving as biological indicator for quickly estimating ionization radiation dosage
A technology of ionizing radiation and biological indicators, which is applied in the field of ionizing radiation biological dose estimation, can solve the problems of complex sampling and long detection cycle, and achieve the effect of simple detection operation, short detection time and simple detection method
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Embodiment 1
[0029] Implementation Example 1: Establishment of Mouse Irradiation Model and Sample Collection
[0030] Experimental animals: male C57BL / 6 mice, 7 weeks old, weighing 19±1g, purchased from Shanghai Slack Experimental Animal Co., Ltd., animal quality certificate number: 2007000534826, experimental animal production license number SCXK (Shanghai) 2007- 0005.
[0031] Serum preparation: Mice were irradiated at 60 In the cobalt source room, 80 mice were randomly divided into four groups, 20 in each group: 1 group in the control group and 4 groups in the radiation group. The mice in the radiation group received 1Gy, 3Gy, 5Gy and 7Gy total body irradiation, respectively, with a dose rate of 0.78Gy / min. 24h, 48h, and 72h after irradiation, blood was collected from the mice through the eyeball. The whole blood was clotted at 37°C for 1h, centrifuged at 3000rpm×15min, and the supernatant serum was separated and distributed in centrifuge tubes at 100μl / tube. Aliquoted serum was froz...
Embodiment 2
[0032] Implementation Example 2: Dose-effect relationship of different doses of ionizing radiation on mouse serum Flt-3L concentration
[0033] (1) Detection of Flt-3L content in serum of non-irradiated mice
[0034] Testing materials and methods:
[0035] Cytokine antibody chip detection. The chip name is: Custom Mouse Array chip, customized by RayBiotech. The chip detection operation steps are as follows:
[0036] ① Place the chip bag at room temperature for 30 minutes, tear off the sealing film, mark the chip, and dry it for 2 hours on a clean bench;
[0037] ②Sample preparation: 60μl serum + 60μl diluent, mix well;
[0038] ③Preparation of standard substance: take 10000pg / ml standard solution (std1), three-fold dilution 6 times (std2~7), the lowest concentration is 13.7pg / ml (std7).
[0039] ④ Preparation of negative control: Take 120 μl of diluent (CNTRL)
[0040] ⑤ Chip blocking and sample incubation: add 100 μl blocking solution to each well, block at room tem...
Embodiment 3
[0063] Implementation Example 3: Statistical Analysis and Fitting of Surfaces and Formulas
[0064] It can be seen from Table 1-4 that the sensitivity of Flt-3L to radiation increases with time after irradiation, and the sensitivity increases, that is, with the same radiation dose, the degree of increase in the concentration of Flt-3L increases with time, and statistics In the analysis, the irradiation time and irradiation dose were used as the influencing factors, and the Flt-3L concentration was used as the effect. SASS9.3 was used to analyze the secondary corresponding surface analysis, and the secondary interaction between irradiation time and irradiation dose was analyzed, and the time- The dose-response corresponding surface diagram, the fitting time-dose-response curve is response=191.54-2.65*time-52.58*dose+0.027*time 2 +1.04*dose 2 +3.06*time*dose, R 2 =0.9711, where response is the concentration of Flt-3L, time is the time after irradiation, and dose is the irradia...
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