Epigenetic modification is crucial to keep the self-renewal and the "stemness" states of stem cells, not letting them to differentiate. The actual roles of Histone 3 Lysine 9 dimethylation (H3K9me2) and its methyltransferase G9a in this process are still unclear, especially in cancer stem cells. In our study, we found an interesting observation that most CD133-positive cells were H3K9me2 negative, both in glioma tissues and in cultured cells, although most cancer cells were detected to be H3K9me2 immunopositive. This implied that the G9a-dependent H3K9me2 was one of the crucial barriers of cancer stem cell self-renewal. To test the hypothesis, we examined the loss-of-function and gain-of-function of G9a. We found that bix01294, the selective inhibitor of G9a, can stimulate the sphere formation rate of glioma cancer stem cells, together with increasing Sox2 and CD133 expressions. The increase of CD133-active stem cells was confirmed by flow cytometry. On the other aspect, overexpression of G9a increased the H3K9me2 and decreased the sphere formation rate as well as the CD133 and Sox2 expressions. Since H3K9me2 modification is the major repressive switch, we predict that the repressive H3K9me2 modification may happen at the CD133 promoter regions. By chromatin precipitation assay, we confirmed that the CD133 and Sox2 promoter regions were modified by the H3K9me2. Therefore, we concluded that the G9a-dependent H3K9me2 repression on CD133 and Sox2 was one of the main switches of the self-renewal in glioma cancer stem cells.
Interest has developed in the potential of mulberry (Morus alba), a woody perennial, for revegetating the hydro-fluctuation belt of the Three Gorges Reservoir due to its resistance to water-logging stress. To be useful, the trees must also be able to withstand dry conditions in summer when temperatures can be very high and droughts become severe quickly. Here, we report a study in which mulberry seedlings were grown in a greenhouse under a variety of simulated soil water conditions reflecting potential summer scenarios in the hydro-fluctuation belt of the Three Gorges Reservoir Area. We compared the responses of two pretreatment groups of mulberry seedlings to different levels of drought stress. The pretreatment groups differed with respect to drought hardening: the daily-managed (DM) group had relative soil moisture held constant in the range 70-80 %, while the drought-hardened (DH) group had relative soil moisture held constant at 40-50 %. Following the month-long pretreatment of seedlings, the two groups of young trees (DM and DH) were then respectively subjected to three levels of drought stress for a month: normal watering, moderate drought stress, and severe drought stress. A series of measurements comparing the physiological status of the plants in the two groups were then made, and the following results were obtained: (1) As drought stress increased, the heights, base diameters, root surface areas, photosynthetic rates (Pn), stomatal conductances (Gs), and transpiration rates (Tr) of the mulberry trees in both groups (DM and DH) decreased significantly, while the specific root area and abscisic acid (ABA) contents had increasing trends. Root activity and instantaneous water use efficiency of mulberry trees in both groups (DM and DH) were all raised under drought stress conditions than under normal watering, but the root/shoot ratio and leaf water potential were lowered. (2) At the same level of soil water content, the heights, base diameters, root/shoot ratios, root surface areas, specific root areas, photosynthetic rates (Pn), stomatal conductances (Gs), and transpiration rates (Tr) of the young mulberry trees in the DH were all significantly higher than those of the control group (DM). Leaf water potential, instantaneous water use efficiency, and abscisic acid content of DH were all significantly lower than DM. Under different degrees of drought stress, the growth of mulberry trees will be inhibited, but the trees can respond to the stress by increasing the root absorptive area and enhancing capacity for water retention. Mulberry trees demonstrate strong resistance to drought stress, and furthermore drought resistance can be improved by drought hardening during the seedling stage.
In the present study, the biosynthesis of silver nanoparticles (AgNPs) and their antibacterial activity against gram-positive and gram-negative bacteria were investigated. Glycyrrhizin (GL) was used as a reducing agent and stabilizer to rapidly prepare the AgNPs. The distinctive absorption peak at 419 nm confirmed the formation of GL-reduced AgNPs. The TEM and particle size analysis shows that the prepared GL-reduced AgNPs were mostly circular with good dispersion and a relatively uniform particle size of 35 nm on average. Fourier transform infrared spectroscopy analysis was performed to identify the possible biomolecules in the capping and active stabilization of the GL-reduced AgNPs. The antibacterial activity of the GL-reduced AgNPs was analyzed with the Oxford cup diffusion method and filter paper diffusion method. The experimental results show that these properties endowed the GL-reduced AgNPs with high antibacterial activity against Escherichia coli and Staphylococcus aureus and lay a foundation for the use of colloidal silver in antibacterial applications. The GL-reduced AgNPs also had stronger antibacterial activity than sodium citrate-reduced AgNPs, which indicates the advantages of GL-reduced AgNPs compared with sodium citrate-reduced AgNPs in inducing bacteriostasis. The cytotoxicity of GL-reduced AgNPs on human kidney epithelial 293A (HEK293) cells was evaluated via the MTT assay. The results show that GL-reduced AgNPs had lower toxicity to HEK293 cells than sodium citrate-AgNPs, which indicates that the as-prepared GL-reduced AgNPs are environmentally friendly.
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