Additional file 1 of Homoharringtonine enhances cytarabine-induced apoptosis in acute myeloid leukaemia by regulating the p38 MAPK/H2AX/Mcl-1 axis

Supplementary Material 1

Figures S1 - S5 from Preparation of core–shell structured CaCO₃ microspheres as rapid and recyclable adsorbent for anionic dyes

N₂ adsorption/desorption isotherms and pore-size distribution (inset) of the obtained core-shell structured CaCO₃ MSs; TG analysis of Hesp (a), pure CaCO₃ without Hesp and the obtained core-shell structured CaCO₃ MSs (b).; SEM image of the CaCO₃ MSs after 5 recycling experiments.; N₂ adsorption/desorption isotherms and pore-size distribution (inset) of the CaCO₃ MSs after 5 recycling experiments.; Three-dimensional histogram of the dye adsorption efficiency on the CaCO₃ MSs in 6-10th cycles of the adsorption-desorption

The effect of isoalantolactone on the expression of cell cycle regulators in UM-SCC-10A cells.

<p>(A) Representative pictures for p53, p21 and cyclin D protein expression by western blot analysis. β-actin was used as a control. (B) Isoalantolactone up-regulated p53 and p21 expression, while down-regulating cyclin D expression in a dose-dependent manner. The results are represented as the means ± SEM from three independent experiments with similar results. *<i>P</i><0.05 and **<i>P</i><0.01 compared to the control.</p

The effect of isoalantolactone on the expression of caspase-dependent mitochondrial apoptosis pathway proteins in UM-SCC-10A cells.

<p>(A) Representative images of cytochrome c, Bax, Bcl-2 and caspase 3 protein expression detected by western blot. β-actin was used as a control. (B) The data are represented as the means ± SEM from three independent experiments with similar results. *<i>P</i><0.05 and **<i>P</i><0.01 compared to the control.</p

The chemical structure of isoalantolactone and its growth-inhibiting effect on UM-SCC-10A cells and mouse splenocytes.

<p>(A) The chemical structure of isoalantolactone. (B) UM-SCC-10A cells were pretreated with 0.1% DMSO or various concentrations of isoalantolactone for 24 and 48 h, and cell growth inhibition assays were performed using the MTT method. The data are expressed as the mean ± SEM of three independent experiments. (C) Isoalantolactone-induced morphologic changes in the UM-SCC-10A cells, and these changes were observed using inverted phase contrast microscopy. The control cells adhered well and displayed normal UM-SCC-10A cell morphology. After isoalantolactone treatment, many cytoplasmic vacuoles were observed in the cells. The vacuoles became progressively larger and denser with increasing concentrations of isoalantolactone. (D) Mouse splenocytes were treated with 25 and 50 µM isoalantolactone for 24 h and stained with 0.4% trypan blue, after which they were examined for dead and living cells microscopically. The dead cells stained blue. (E) Cell viability after 25 or 50 µM isoalantolactone treatment in UM-SCC-10A cells and mouse splenocytes for 24 h. The results are the mean ± SEM from three independent experiments. *<i>P</i><0.05 and **<i>P</i><0.01 compared to the control.</p

The effect of isoalantolactone on the MMP in UM-SCC-10A cells.

<p>(A) The MMP of UM-SCC-10A cells treated with isoalantolactone at different concentrations was analyzed by flow cytometry. (B) The loss of the MMP in UM-SCC-10A cells following isoalantolactone treatment in a dose-dependent manner. The data are expressed as the means ± SEM for three independent experiments with similar results. *<i>P</i><0.05 compared to the control.</p

Isoalantolactone-induced apoptosis in UM-SCC-10A cells.

<p>(A) Apoptosis was evaluated using an annexin V-FITC apoptosis detection kit and flow cytometry. The X- and Y-axes represent annexin V-FITC staining and PI, respectively. The representative pictures are from UM-SCC-10A cells incubated with different concentrations of isoalantolactone (25 and 50 µM) or caspase inhibitor (Z-VAD-FMK 50 µM). (B) Isoalantolactone induced apoptosis in the UM-SCC-10A cells in a dose-dependent manner. Z-VAD-FMK markedly reduced apoptosis in UM-SCC-10A cells treated with high-dose isoalantolactone. The data are expressed as the means ± SEM of three independent experiments with the similar results. *<i>P</i><0.05 and **<i>P</i><0.01 compared to the control. (C) The morphological nuclear changes in UM-SCC-10A cells treated with isoalantolactone at different concentrations. The cells were stained with Hoechst33258 for 30 min in the dark to examine the cleaved nuclei, which is a sign of apoptosis.</p

The effect of isoalantolactone on the cell cycle in UM-SCC-10A cells.

<p>(A) The DNA content in each cell cycle phase in UM-SCC-10A cells was analyzed by flow cytometry. The representative histograms are from UM-SCC-10A cells incubated with different concentration of isoalantolactone (25 and 50 µM) or caspase inhibitor (Z-VAD-FMK50 µM). (B) Isoalantolactone treatment induced a dose-dependent increase in the proportion of cells in the G1 phase and a decrease in cells in the S and G2 phases compared to the control. Z-VAD-FMK treatment did not prevent cell cycle arrest following high-dose isoalantolactone treatment. The results are represented as the mean ± SEM for three independent experiments with similar results. *<i>P</i><0.05 and **<i>P</i><0.01 compared to the control.</p

Nanoscale Glutathione-Functionalized Bone Marrow Mesenchymal Stem Cell-Derived Exosomes Loaded with Metformin for the Treatment of Spinal Cord Injury

Spinal cord injury (SCI) is a central nervous system disease with a high disability. Immune activation of microglia cells can be induced, and the activated microglia cells are mainly divided into two different subtypes, namely, proinflammatory phenotype (M1) and anti-inflammatory phenotype (M2). Regulating the transformation of microglial subtypes is the key to alleviating inflammation. However, because of the blood–spinal cord barrier (BSCB), most drugs cannot reach the target site and give a full effect. Therefore, the purpose of this study was to design a nanoscale glutathione-functionalized bone marrow mesenchymal stem cell-derived exosome (Exos-GSH) as a delivery carrier for metformin. Using Exos-GSH’s ability to cross BSCB, metformin can be efficiently delivered to the injured spinal cord tissue and taken up by neurons and microglia cells at the injured site. Exos-GSH loading metformin (Exos-Met-GSH) had a particle size of about 154 ± 17 nm, and the encapsulation rate was 87.49 ± 3.36%. In vitro and in vivo experiments showed that Exos-Met-GSH could exert good anti-inflammatory effects by inducing the polarization of microglia from the M1 phenotype to the M2 phenotype. In addition, Exos-Met-GSH can also protect mitochondria by relieving the oxidative stress of neurons, thus inhibiting neuronal apoptosis. Finally, Exos-Met-GSH can protect nerve cells through anti-inflammatory, antioxidant stress, and inhibition of apoptosis, thus promoting the recovery of motor function in SCI mice, which is a potential drug for SCI treatment

Macrophage Membrane-Coated MnO₂ Nanoparticles Provide Neuroprotection by Reducing Oxidative Stress after Spinal Cord Injury

Secondary injury following spinal cord injury (SCI) results in a large production of reactive oxygen species (ROS) (e.g., H2O2) in the spinal cord microenvironment, which then leads to an excessive burst of inflammation and ultimately neuronal death. In this study, we prepared manganese dioxide (MnO2) nanoparticles coated by macrophage membranes, named M@MnO2, to cope with early ROS bursts in the SCI microenvironment. The biosafety and targeting ability of the MnO2 nanoparticles were improved through the macrophage membranes. Successful preparation of M@MnO2 was verified by transmission electron microscopy, Western blot, and dynamic light scattering. Small animal imaging showed that M@MnO2 accumulated in large quantities at the site of SCI. In the early stages of SCI, M@MnO2 effectively reduced the ROS content, as well as the hypoxia-inducible factor 1α (HIF-1α) content, malondialdehyde content, and superoxide anion content caused by ROS, further leading to a decrease in some of the proteins associated with inflammation at the site of SCI (CD11b, CD86, COX2, IL-1β, and iNOS), ultimately achieving neuroprotection and recovery of motor function