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Objective To investigate the protective effect of hydrogen-rich water (HRW) on radiation-induced hematopoietic stem and progenitor cells (HSPCs) injury.Methods Totally 32 C57BL/6 mice were randomly divided into four groups with 8 mice in each group,including control,HRW,radiation and radiation + HRW.Mice in HRW and radiation + HRW groups received 0.5 ml hydrogen-rich water per day by intragastric administration 5 min before irradiation until 7 d post-irradiation.Mice in other groups received 0.5 ml distilled water.Mice in radiation and radiation + HRW group were irradiated with 2 Gy of total body irradiation.Bone marrow cells were isolated at 15 d post-irradiation,and LSK cells were examined for the percentage of hematopoietic stem and progenitor cells,the ability of colony formation and reconstitution,reactive oxygen species (ROS) levels and cell apoptosis.Results Compared with radiation group,the percentages of hematopoietic progenitor cells and LSK cells,colony number of bone marrow cells were significantly increased in radiation + HRW group (t =-4.935,-7.898,5.488,P ＜ 0.05).An elevation of donor chimerism was also found in recipient mice administered HRW after competitive bone marrow transplantation (t =-12.769,P ＜ 0.05).Compared with radiation group,the ROS levels and cell apoptosis in LSK cells were significantly decreased (t =4.380,3.954,P ＜ 0.05).Conclusions Hydrogen-rich water exhibited a protective effect on radiation-induced HSPCs injury.
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Objective To investigate the protective effect of theaflavins on thymus injury caused by total body irradiation (TBI).Methods Twenty-five C57BL/6 mice were randomly divided into 5 groups:control group,4 Gy TBI group,4 Gy TBI + 25 mg/kg theaflavins group,4 Gy TBI + 50 mg/kg theaflavins group and 4 Gy TBI + 100 mg/kg theaflavins group.Thymus index and total number of thymocytes were detected at the 14th d post-irradiation to determine the optimal dose of theaflavins.According to this optimal dose,32 C57BL/6 mice were randomly divided into 4 groups:control group,theaflavins group,4 Gy TBI group and 4 Gy TBI + theaflavins group.Thymus histomorphology,CD4CD8 T cell subsets,and reactive oxygen species (ROS) in thymocytes were examined at the 14th d post-irradiation.Results The irradiated thymus exhibited decreased thymus index and total number of thymocytes (P ＜ 0.05),aberrant histomorphology and T cell subsets (P ＜ 0.05),and increased ROS level in thymocytes (P ＜ 0.05).Compared with 4 Gy TBI group,the thymus index and total number of thymocytes were significantly increased in 4 Gy TBI + 50 mg/kg theaflavins group (P ＜ 0.05).The total number of thymocytes was significantly higher in 4 Gy TBI + 50 mg/kg theaflavins group than that in 4 Gy TBI + 25 mg/kg theaflavins group (P ＜ 0.05).Therefore,50 mg/kg theaflavins was chosen as the optimal dose for subsequent experiments.Moreover,the aberrant histomorphology of irradiated thymus was alleviated by theaflavins.A decline in the percentage of CD4-CD8-T cells and an elevation of CD4+CD8-and CD4+CD8+ T cells were found in irradiated mice administered with theaflavins (P ＜ 0.05).Compared with 4 Gy TBI group,the ROS level was significantly decreased in 4 Gy TBI + theaflavins group (P ＜ 0.05).Conclusion Theaflavins exhibits a protective effect on radiation-induced thymus injury.
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Objective To observe the protective effect of anthocyanin on irradiation induced bone marrow c-kit positive cell injury, and further explore its possible mechanism. Methods Mouse bone marrow c-kit positive cells were collected by cell sorting method. There were 2 groups: control group and anthocyanin group, which were sub-divided into three groups and received 0 Gy, 1 Gy and 4 Gy irradiation respectively. The control group was added 700μL cell suspension and an equal volume of serum-free hematopoietic stem/progenitor cell culture medium. The 2 × 10-5 mol/L anthocyanin was co-cultured with mouse bone marrow c-kit positive cells of anthocyanin group half an hour before irradiation exposure, then cells were cultured for 18 hours under the conventional culture conditions (37℃,5%CO2). Mouse c-kit positive cell viability was measured by bioluminescence, and which was reflected by relative light units (RLU). The ability of colony-forming units was reflected by CFU-GM. The reactive oxygen species (ROS) level and mean fluorescence intensity (MFI) ofγ-H2AX were detected by flow cytometry. Results Compared to un-irradiated control group, the cell viability and the number of CFU-GM were decreased significantly, while the ROS level and MFI ofγ-H2AX were increased in c-kit positive cells irradiated with 1 Gy and 4 Gy (P<0.05). Compared to 1 Gy and 4 Gy irradiation groups, c-kit positive cell viability and the number of CFU-GM were increased, the ROS level and MFI of γ-H2AX were decreased in anthocyanin group (P < 0.05). Conclusion Anthocyanin exhibits a promising protective effect on radiation-induced bone marrow c-kit positive cell injury, which may be related to the alleviating ROS and DNA damage in bone marrow cells.
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Objective To compare the advantages and disadvantages of different immunohistochemical methods in detecting isocitrate dehydrogenase-1 (IDH 1) mutations in gliomas,and to optimize the processes these detection.Methods One hundred and thirty-eighty glioma specimens,collected and conformed by pathology in our hospital from January 2013 to December 2013,were used in our study,including 18 of WHO grade Ⅰ,49 of WHO grade Ⅱ,24 of WHO grade Ⅲ and 47 of WHO grade Ⅳ.Manual immunohistochemical method and automatic immunohistochemical instrument were used to detect the IDH1 mutation.PCR-high resolution melting curve analysis (PCR-HRM) was used to verify the above results.Results There were 65.9％ positive specimens those had IDH1 positive tumor cells higher than 75％,and 70.7％ positive specimens those were strong staining.Manual immunohistochemical method enjoyed advantages as clean background,clearness and easy reading,and no interpretation difficulty or false-positive result were noted with this method;while automatic immunohistochemical instrument enjoyed dark background,which led to interpretation difficulty or false-positive result;the results of IDH1 staining had significant differences between and automatic immunohistochemical instrument (x2=22.042,P=0.000).The positive detection rate of automatic immunohistochemical instrument was significantly higher than that of manual immunohistochemical method,and the results of IDH1 detection had no significant difference between manual immunohistochemical method and PCR-HRM (x2=0.800,P=0.371).Conclusions The results of IDH1 detection by manual immunohistochemical method are more accurate than that of immunohistochemical instrument.IDH1 gene mutation only has a relationship with the number of positive tumor cells,and not the staining intensity.The specimen can be considered to IDH1 gene mutation when the positive cells are more than 5％.