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Objective:To study the influence of circular RNA hsa_circZDHHC21_004 on the proliferation of human small intestinal epithelial cells HIEC-6 after 60Co γ-rays exposure. Methods:HIEC-6 cells were exposed to 60Co γ-rays at 0, 5, 10, and 15 Gy with a dose rate of 1 Gy/min. The expression level of hsa_circZDHHC21_004 in the irradiated HIEC-6 cell was detected. Hsa_circZDHHC21_004 was knocked-down to investigate the influences of hsa_circZDHHC21_004 on the proliferation of irradiated HIEC-6 cells by CCK-8 assay and colony formation assay. Results:The expression level of hsa_circZDHHC21_004 in HIEC-6 cells was upregulated by (1.00±0.24), (1.34±0.28), (1.85±0.31), and (2.80±0.64) times of control after 0, 5, 10, and 15 Gy irradiation, respectively and there were significant difference between 10 or 15 Gy group and 0 Gy group ( F=10.86, P=0.008). Knockdown of hsa_circZDHHC21_004 significantly increased the proliferation rate of HIEC-6 cells at 24, 48, and 72 h after 10 Gy irradiation compared with non-irradiated control ( t=-6.25, -5.83, -7.75, P < 0.001). Under 2 and 5 Gy irradiation, the clone formation rates of the hsa_circZDHHC21_004 knockdown cells were significantly higher than those of the control ( t=-7.45, -8.83, P<0.01). Conclusions:Hsa_circZDHHC21_004 is increased after irradiation and influenced the proliferation of irradiated HIEC-6 cells.
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Objective:To investigate the metabolite changes in rat plasma after total body irradiation (TBI) and to explore dose classification based on radiation sensitive metabolites.Methods:The differential metabolites induced by radiation were screened and verified by metabolomics. In the discovery stage, 50 SD rats were irradiated with 0, 1, 2, 3, 5 and 8 Gy of 60Co γ-rays. In the verification stage, 25 rats were irradiated with 0, 0.5, 2.5, 4 and 6 Gy. Peripheral blood samples were collected 4 h after irradiation, and plasma was separated. Radiation-induced differential metabolites were identified and their concentrations were determined. Receiver operating characteristic (ROC) curve of the differential metabolites was used to classify dose range. Results:In the discovery stage, 8 radiation-induced differential metabolites in rat plasma were identified and four of them (cytosine, L-hexylcarnitine, Linoelaidylcarnitine and L-palmitylcarnitine) were upregulated, which was confirmed in the verification stage. The area under the curve (AUC) for the specific dose was >0.75. After combining these four metabolites, the AUC value to classify the radiation dose of 0 Gy versus >0 Gy, <2 Gy versus ≥2 Gy, <5 Gy versus ≥5 Gy were 0.96, 1 and 0.94, respectively.Conclusions:The metabolites in rat plasma changed significantly at 4 h after TBI, where 8 differential metabolites were identified. Cytosine, L-hexylcarnitine, linoelaidylcarnitine and L-palmiylcarnitine were stably over-expressed in the plasma after irradiation. The combination of these four compounds had high classification accuracy and thus may applicable as radiation sensitive biomarkers for dose classification.
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Objective:To screen radiosensitive lipid metabolites in rat plasma and analyze their metabolic pathways in order to provide scientific basis for radiation damage biomarker.Methods:The whole body irradiation of 60Co γ rays was performed to rats with different doses of 0, 1, 3 and 5 Gy. The changes of lipids in plasma were detected by untargeted lipidomics method based on liquid chromatography coupled mass spectrometry. Results:Twenty plasma lipids were identified as the potential radiosensitive biomarkers at 7 days after irradiation, including 13 over-expressed lipids and 7 down-expressed lipids, where 12 lipids well responded to radiation doses.Conclusions:Lipid metabolites in rat plasma are significantly changed after exposure to γ rays, and the metabolic pathways of sphingolipid, glycerophospholipid and glycosylphosphatidylinositol (GPI) are significantly enriched.
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Objective:To explore the effects of ultraviolet B (UVB) on the premature senescence of human immortalized keratinocytes HaCaT cells and the possible underlying molecular mechanism.Methods:HaCaT cells were exposed with UVB of different doses (20, 50, 80 and 100 mJ/cm 2). At 72 h after exposure, cellular morphology was observed by Giemsa staining, cell proliferation was detected by clone formation assay, and the proportion of premature senescence cells was detected by β-galactosidase staining. The number change of lysosomes was detected by Lyso-Tracker Red fluorescence probe at 24, 48 and 72 h after exposure. Cell migration was measured by scratch test at 24 h and 48 h after exposure. The protein expressions of p53 and p16 related to premature senescence were detected by Western blot assay at 72 h after exposure. Results:After UVB exposure, HaCaT cells showed a premature senescence phenotype. At 72 h after exposure, the cell volume increased ( F=115.18, P<0.05), the cell proliferation ability decreased ( F=410.32, P<0.05), the activity of β-galactosidase increased ( F=16.31, P<0.05), and the expressions of P53 and P16 increased. In addition, the number of lysosomes increased at 24, 48, and 72 h after exposure ( F=17.65, 38.36, 13.66, P<0.05), and cell migration capacity was inhibited at 24 and 48 h after exposure ( F=8.21, 11.48, P<0.05). Conclusions:UVB exposure can induce premature senescence of HaCaT cells by increasing the expression of p53 and p16 proteins.
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Objective:To screen the indicators of retrospective dose estimation, based on 5 cytogenetic methods to assess the victim followed-up at 4 year after 192Ir radiation accident in Nanjing. Methods:The chromosome aberration (dic + r) assay, cytokinesis block micronucleus (MN) and nucleoplasmic bridge (NPB) assay, fluorescence in situ hybridization (FISH)-based and G banding-based translocation analysis were used to retrospective biological dose estimation. Results:The estimated doses of FISH-based and G banding -based analysis were 1.45 and 1.21 Gy respectively, which was similar to the biological dose estimated short time after the accident. However, the estimated doses by chromosome aberration, micronucleus and nucleoplasmic bridge method were 0.56, 0.45 and 0.41 Gy respectively, which were lower than the corresponding biodose. Correction factors were used to adjust the biodose.Conclusions:In the 4th years after exposure, the estimated biological doses by FISH-based and G banding-based translocation were consistent with the biodose.Therefore, the two methods were suitable for retrospective dose estimation, while correction factors should be considered in chromosome aberration method for retrospective dose estimation.
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Objective To study the effects of sex,age,length of service,type of work and annual effective radiation dose on nucleoplasmic bridge (NPB) in the peripheral blood lymphocytes of radiation workers.Methods The peripheral blood samples of 100 radiation workers in Henan province were collected and the NPB in peripheral blood lymphocytes were measured by CBMN assay.The frequencies of NPB formation and NPB-containing cells were calculated,and the effects of various factors on NPB incidence were analyzed statistically.Results The NPB frequency in radiation workers was higher than that in healthy people (Z=-8.123,P＜0.01).Except for sex,the factors of age,length of service,type of work and annual effective dose had significant influences on NPB (x2=7.202-45.571,P＜0.05).Conclusions NPB reflects the effect of low-dose long-term chronic irradiation on the occupational radiation workers.
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Objective@#To study the effects of sex, age, length of service, type of work and annual effective radiation dose on nucleoplasmic bridge (NPB) in the peripheral blood lymphocytes of radiation workers.@*Methods@#The peripheral blood samples of 100 radiation workers in Henan province were collected and the NPB in peripheral blood lymphocytes were measured by CBMN assay. The frequencies of NPB formation and NPB-containing cells were calculated, and the effects of various factors on NPB incidence were analyzed statistically.@*Results@#The NPB frequency in radiation workers was higher than that in healthy people (Z=-8.123, P<0.01). Except for sex, the factors of age, length of service, type of work and annual effective dose had significant influences on NPB (χ2= 7.202-45.571, P<0.05).@*Conclusions@#NPB reflects the effect of low-dose long-term chronic irradiation on the occupational radiation workers.
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OBJECTIVE: To explore the dose-effect of X-ray irradiation and chromosome aberrations in human peripheral blood cells,and establish a dose-response curve of dicentric and ring(dic+r)aberration induced by X-ray irradiation.METHODS: Human peripheral blood samples were collected from three healthy individuals and were exposed to X-ray at the doses of 0.00,0.25,0.50,0.75,1.00,2.00,3.00,4.00 and 5.00 Gy in vitro.The dose rate was 1.158 Gy/min.The blood cells were harvested after routine culture,and the chromosome preparation was carried out.The dicentrics and rings in metaphase cells were counted under microscope,and a dose-response curve was fitted by using the software of CABAS.Dose estimation was performed according to the curve from two blind samples.RESULTS: Aberration of dic+r increased with irradiation doses in the range of 0.00-5.00 Gy(P<0.01).The dose-response relationship followed a linear-quadratic equation:■,where■ is the yield of dic+r,and D is the irradiated dose.The estimated doses of the two blind samples were in accordance with the actual doses.CONCLUSION: The dose-response curve and mathematical model of chromosome aberration following exposure to 0.00-5.00 Gy X-ray irradiation is established in this study provide a reliable method for the accurate dose estimation.
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Objective To explore the influences of the final concentration and adding time of Cytochalasin-B (Cyt-B) on radiation-induced nucleoplasmic bridges (NPB) in cytokinesis-block assay.Methods Hunan peripheral blood samples were divided into 5 final concentration groups (group 2,4,6,8,10 μg/ml) according to different final concentrations of Cyt-B.Moreover,blood samples were divided into 4 adding time groups (group 0,28,40,44 h) according to different adding times of Cyt-B.Blood samples were irradiated with 0 (sham irradiation) and 2 Gy 60Co-rays in vitro,at a dose rate of 1 Gy/min.A cytokinesis-block assay was carried out to prepare NPB samples.The percentages of mononucleated,binucleated and multinucleated cells,as well as the frequencies of NPB and micronucleus (MN) in binucleated cells were analyzed using an optical microscope.Results Nuclear division index (NDI) and the percentages of binucleated cells increased with increased concentration of Cyt-B,and decreased with delayed adding time of Cyt-B (except group 0 h) in both final concentration groups and adding time groups.After exposed to 2 Gy,NPB frequencies were no significant difference (except group 0 h).MN frequencies had the trend of decreased with the increased concentration of Cyt-B,but no significant difference with adding time of Cyt-B.Conclusions In cytokinesis-block assay,different final concentration and adding time of Cyt-B may induce to the variation of NPB frequencies,but there was no significant difference.Appropriate increased final concentration or ahead adding time of Cyt-B can increase the percentage of binucleated cells that help to improve the efficiency of analysis.