The Role of Nanoparticles-Mediated Ligustrazine in the Treatment of Knee Osteoarthritis and Its Effect on Matrix Metalloproteinases and Upstream NF-κB Signaling Pathway in Knee Osteoarthritis.

Knee osteoarthritis (KOA) is a joint degenerative arthropathy, characterized by cartilage degeneration of knee joint. Ligustrazine is an effective component of traditional Chinese medicine chuanqiong. It is reported that ligustrazine is used as a kind of anti-inflammatory medicine in folk prescription, especially in the treatment of knee osteoarthritis. The study is aimed to study the therapeutic effect of ligustrazine mediated by nanoparticle on knee osteoarthritis and its impact on MMPs and upstream NF-κB signaling pathway in synovial fluid. Nanoparticle-mediated system is a kind of nano traditional Chinese medicine preparation, which is made by taking nanoparticle and combining with the effective components, effective parts, raw materials, and their compounds in a certain way. We found that the combination of nanoparticle and ligustrazine can improve its bioavailability and targeting, reduce the adverse reactions in the treatment of knee osteoarthritis. The ligustrazine mediated by nanoparticle can effectively alleviate knee osteoarthritis by reducing the level of MMPs in synovial fluid and the expression of NF-κB in upstream NF-κB signaling pathway.

Shikonin overcomes drug resistance and induces necroptosis by regulating the miR-92a-1-5p/MLKL axis in chronic myeloid leukemia.

Development of resistance to tyrosine kinase inhibitors (TKIs) targeting the BCR/ABL fusion protein represents a major challenge in the treatment of chronic myeloid leukemia (CML). Since apoptosis resistance is the fundamental mechanism impeding TKIs' therapeutic effects, alternative approaches that induce nonapoptotic cell death are being pursued to treat TKI-resistant CML. Induction of necroptosis, a distinct, caspase-independent form of programmed cell death, may be a valuable strategy in this respect. The present study shows that shikonin, an herbal compound used in traditional Chinese medicine, overcomes TKI resistance in BCR/ABL-positive CML cells by inducing necroptosis via activation of RIPK1/RIPK3/MLKL signaling. This effect occurs both in vitro and in vivo and involves downregulation of miR-92a-1-5p, a poor-prognosis marker frequently overexpressed in leukemia patients. Based on gene expression experiments, we conclude that miR-92a-1-5p promotes CML progression by inhibiting MLKL expression. Accordingly, we show that antagomiR-mediated in vivo inhibition of miR-92a-1-5p reduces the growth of CML tumors in mice through necroptosis induction. Our research suggests that therapies that relieve MLKL suppression by targeting miR-92a-1-5p may represent a useful strategy to treat TKI-refractory CML.

Modeling of the static batch desorption and dynamic column elution of geniposidic acid from a porous anion-exchanger.

For several decades, plenty of iridoid glycosides including geniposide (GS) and geniposidic acid (GSA) in the gardenia yellow pigment extraction waste water was not recovered effectively. This study is aimed to supply an efficient GSA recycling route. In this study, a model incorporating a superficial desorption rate constant was applied to the batch GSA desorption process, i.e., recycling, for verification. Then, the model was further developed to research the feasibility in dynamic column elutions simulation through porous uniform media. The simulation approach was done by coupling velocity field and mass transfer equations using COMSOL Multiphysics™ Finite element method, with appropriate mesh refinement was employed to solve the equation system. The HCl solutions ranging from 0.03 mol/L to 0.06 mol/L were used to desorb/elute the GSA from a presaturated polymeric porous anionic resin D08. Good results were accomplished in terms of ion exchange desorption rate and GSA recovery. The pore diffusion model (PDM) considering counter ion was established to describe the desorption/elution kinetics in the batch/column experiment. By the least square fitting method, the superficial desorption rate constant Kd of GSA/HCl reaction on the ion-exchange sites of porous resin was fitted to 0.116 L/(mol s). Subsequently, this value was sequentially applied in the simulation of the dynamic elution process. The individual pore diffusion coefficients for GSA and Cl- were estimated to be 5.07 × 10-10 and 1.77 × 10-9 m2/s, respectively. In order to validate the simulation feasibility of this pore diffusion model to a dynamic column elution process, the effects of HCl concentration, flow rate and column's height/diameter ratio on the column performance were investigated systematically. The results from this work should serve as motivation for further experimental and theoretical study in the scaling-up of GSA purification process. Finally, repeated adsorption-elution column cycles were simulated by the PDM model well.

Shikonin exerts antitumor activity in Burkitt's lymphoma by inhibiting C-MYC and PI3K/AKT/mTOR pathway and acts synergistically with doxorubicin.

Burkitt's lymphoma (BL) is a highly aggressive malignancy molecularly characterized by deregulation of the C-MYC proto-oncogene. Recently, it has been confirmed that phosphatidylinositol-3-kinase (PI3K) pathway activation is a crucial element in the malignant transformation of the B cells in BL. Despite the better outcome of adults with BL treated with high-intensity chemotherapy regimens, the overall survival rate for patients older than 60 years remains dismal. Shikonin, a natural naphthoquinone derived from Chinese herbal medicine plant, has the potential to induce cell death in a series of human cancer. In the present study, we investigated the effect and molecular mechanisms of Shikonin in treatment with BL. Shikonin suppressed cellular proliferation and induced caspase-dependent apoptosis in BL cells. Inhibition of C-MYC and suppression of PI3K/AKT/mTOR pathway played critical roles in SHK-induced apoptosis in BL both in vitro and in vivo. Besides, Shikonin potentiated doxorubicin-induced growth inhibition and apoptosis in vitro. Furthermore, the growth of a subcutaneous xenograft tumor model of BL was significantly inhibited by shikonin. Importantly, we did not find the effect of shikonin on liver function in mice. In summary, these data suggest that shikonin may be an encouraging chemotherapeutic agent in the clinical treatment of BL.

ERG11 Gene Mutations and MDR1 Upregulation Confer Pan-Azole Resistance in Candida tropicalis Causing Disseminated Candidiasis in an Acute Lymphoblastic Leukemia Patient on Posaconazole Prophylaxis.

In this study, we present a rare case of fatal breakthrough Candida tropicalis infection in a patient with acute lymphoblastic leukemia (ALL) while on posaconazole prophylaxis. Then, we explore the mechanisms underlying azole resistance by focusing on enhanced efflux pumps and changes in the azole target enzyme Erg11p, which was encoded by the ERG11 gene. Our study demonstrates that Y132C substitution of Erg11p combined with MDR1 overexpression may be the pan-azole resistance mechanisms in Candida tropicalis.

Total coumarins of Hedyotis diffusa induces apoptosis of myelodysplastic syndrome SKM-1 cells by activation of caspases and inhibition of PI3K/Akt pathway proteins.

ETHNOPHARMACOLOGICAL RELEVANCE: Hedyotis diffusa is an ethno-medicine used for anti-cancer treatment in the clinic of traditional Chinese medicine (TCM). The total coumarins of Hedyotis diffusa (TCHD) was a selected extract with observed antiproliferative activity, which has not been tested in treatment of myelodysplastic syndromes (MDS) or acute myeloid leukemia (AML). AIM OF THE STUDY: This study aimed to evaluate the apoptosis-inducing effect of TCHD on human MDS cell line (SKM-1) and explore its action mechanism in association with caspase family and PI3K/Akt signaling pathway. MATERIALS AND METHODS: The chemical constituents and total coumarins content of TCHD were determined by High Performance Liquid Chromatography-tandem mass spectrometry (HPLC-MS/MS) and UV-vis spectrophotometry, respectively. MTT assay, Hoechst 33258 staining, and Annexin V-FITC/PI double labeling were applied to evaluate TCHD's efficacy on SKM-1 cells. Western blot analysis was also used to clarify the action mechanism of TCHD on protein expression level. RESULTS: Two compounds, p-coumaric acid and E-6-O-p-coumaroyl scandoside methyl ester, were identified in TCHD, and its total coumarins content reached 87.4%. By MTT assay, apoptosis-inducing effect of TCHD on SKM-1 cells was found in a dose-dependent manner after 24-48h treatment, with IC50 values of 104.48µg/ml and 100.66µg/ml, respectively. Morphological and flow cytometry observation also confirmed such effect of TCHD. Western blot analysis clarified its action mechanism associating with the activation of caspases and inhibition of PI3K/Akt pathway proteins. CONCLUSIONS: This is the first report regarding the apoptosis-inducing efficacy and mechanism of TCHD on SKM-1 cells, providing a promising candidate of TCM for MDS and AML therapy with fewer side effects.

Pure total flavonoids from Citrus paradisi Macfadyen act synergistically with arsenic trioxide in inducing apoptosis of Kasumi-1 leukemia cells in vitro.

To investigate the potential effects of pure total flavonoid compounds (PTFCs) from Citrus paradisi Macfadyen separately or combined with arsenic trioxide on the proliferation of human myeloid leukemia cells and the mechanisms underlying the action of PTFCs. The effects of PTFCs separately or combined with arsenic trioxide on the proliferation and apoptosis of leukemia cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), fluorescence microscopy, and flow cytometry. Their effects on the expression levels of apoptosis-related regulators were determined by Western blot assay. PTFCs combined with arsenic trioxide significantly inhibited the growth of Kasumi-1 cells, and apoptosis was confirmed by flow cytometry analysis. Hoechst 33258 staining showed more significant morphological changes and more apoptosis following the combined treatment. Western blots showed changes in the expression of genes for poly ADP-ribose polymerase (PARP), caspase 3/9, and P65. The results indicated that PTFCs separately or combined with arsenic trioxide inhibited proliferation of leukemia cells in vitro and induced their apoptosis by modulating the expression of apoptosis-related regulator genes.

Matrine and CYC116 synergistically inhibit growth and induce apoptosis in multiple myeloma cells.

OBJECTIVE: To investigate whether CYC116 can potentiate matrine-dependent growth inhibition and apoptosis in multiple myeloma (MM) cells. METHODS: The dose response relationship of matrine to dexamethasone-resistant and dexamethasone-sensitive MM cells was first established. Myeloma RPMI8226 cells were treated with matrine alone or combined with CYC116 for 24 h. Cell proliferation was measured using an MTT assay and apoptosis induction was evaluated by flow cytometry. Activation of the caspase pathway and expression of apoptosis regulator proteins were detected by Western blotting. RESULTS: Matrine significantly induced growth arrest and apoptosis in both drug-resistant and drug-sensitive MM cells. Treatment with the combination of matrine and CYC116 had a stronger cytotoxic effect on MM cells than did single drug treatments. Enhanced apoptosis observed following the combined treatment of matrine and CYC116 was associated with higher levels of activation of caspase-9, caspase-3, and poly adenosine diphosphate ribose polymerase (PARP) and down-regulation of the anti-apoptotic proteins Bcl-2 and Mcl-1 and the signaling proteins p-Akt and nuclear factor κB (NF-κB). CONCLUSION: CYC116 enhances the growth inhibitory and apoptotic effects of matrine on MM cells.

[Homoharringtonine combined arsenic trioxide induced apoptosis in human multiple myeloma cell line RPMI 8226: an experimental research].

OBJECTIVE: To clarify the effects and mechanisms of homoharringtonine (HHT) monomer therapy or combination therapy with arsenic trioxide (ATO) on human multiple myeloma (MM) cell line RPMI 8226 in in vitro researches. METHODS: Effects of HHT, ATO, and HHT combined ATO on the growth of MM cell line RPMI 8226 were detected using MTT assay. The morphological changes of cell apoptosis were detected by Hoechst staining. The early apoptosis rate was detected using flow cytometry. Expressions of Caspase-3, Caspase-9, poly-ADP-ribose polymerase (PARP), Bcl-2, Mcl-1, Bcl-xl, and AKT protein were detected by Western blot. RESULTS: HHT and ATO inhibited the proliferation of RPM1 8226 cell line in a time- and dose-dependent manner (P < 0.05). Synergistic effects was shown in the combination group (Cl < 1). HHT and ATO induced the apoptosis of RPMI 8226 in a dose-dependent manner with typical morphological changes of apoptosis and higher early stage apoptosis rate. The enhancement in apoptotic induction was seen when two agents were combined. HHT activated expressions of Caspase-3 and PARP in a dose dependent manner at 24 h. HHT at 40 ng/mL and ATO at 8.5 micromol/L could significantly activate expressions of Caspase-3 and Caspase-9, and down-regulate expressions of anti-apoptotic proteins Bcl-xl and Mcl-1. In addition, the combination therapy of HHT at 40 ng/mL and ATO at 8.5 micromol/L inhibited phosphorylation of AKT in a time-dependent manner. CONCLUSION: HTT, ATO, and combination therapy of HHT and ATO induced the apoptosis of RPMI 8226 cell line possibly through activating Caspase pathways, regulating expressions of Bcl-2 families, and inhibiting phosphorylation of AKT.

Arsenic trioxide-enhanced, matrine-induced apoptosis in multiple myeloma cell lines.

Matrine and arsenic trioxide are monomers used in traditional Chinese medicine possessing anti-myeloma activities. In this study, we evaluated the effects and mechanisms of matrine, arsenic trioxide, and their combination therapy on the proliferation and apoptosis of the myeloma cell lines RPMI8226 and U266. The effects of growth inhibition were measured by MTT, and apoptotic cells were analyzed by Hoechst 33258 staining and flow cytometry. The levels of caspase-3, poly (ADP-ribose) polymerase (a DNA repair enzyme), Bcl-2 and survivin (antiapoptotic signaling proteins), Bim (a proapoptotic signaling protein), total AKT, and phosphorylated AKT were evaluated by Western blot. Matrine significantly inhibited proliferation of RPMI8226 and U266 cell lines in a dose- and time-dependent manner with an IC50 at 24 h of 2.25 g/L and 2.18 g/L, and at 48 h of 1.64 g/L and 1.58 g/L, respectively. Arsenic trioxide also displayed a dose- and time-dependent inhibition of growth of multiple myeloma cell lines, and synergistic effects occurred when the two were combined. Matrine (0.5, 1.0 g/L) and arsenic trioxide (2, 4 ug/mL) induced the apoptosis of myeloma cells; more early-stage apoptotic cells were seen with the combination therapy (matrine 0.5 g/L plus arsenic trioxide 2 ug/mL and matrine 1.0 g/L plus arsenic trioxide 4 ug/mL). Activation of caspase-3 and poly (ADP-ribose) polymerase, upregulation of Bim expression, downregulation of Bcl-2, survivin expression, as well as inhibition of phosphorylated AKT related to matrine (0, 0.25, 0.5, 1.0, and 2.0 g/L)-mediated apoptosis, and the effects were enhanced when arsenic trioxide (8 ug/mL) was combined with matrine (1.0 g/L). In conclusion, matrine displayed anti-myeloma effects through apoptotic induction, and arsenic trioxide had synergistic effects with matrine enhancing matrine-induced apoptosis.

Heat shock protein 90 inhibition results in altered downstream signaling of mutant KIT and exerts synergistic effects on Kasumi-1 cells when combining with histone deacetylase inhibitor.

KIT mutations may be associated with a poor prognosis in t(8;21) AML. Heat shock protein 90 (Hsp90) is a molecular chaperone frequently used by cancer cells to stabilize mutant oncoproteins. Inhibition of Hsp90 by 17-allylamino-17-demethoxygeldanamycin (17-AAG) disrupted downstream signaling pathways of mutant KIT in Kasumi-1 cells. AML1-ETO fusion gene and mutated KIT act as "two-hit" factors in Kasumi-1 cells. Histone deacetylation (HDAC) inhibitors sodium phenylbutyrate (PB) and valproic acid (VPA) block AML1-ETO. Co-treatment with 17-AAG and PB or 17-AAG and VPA resulted in a synergistic effect in Kasumi-1 cells. Our results confirmed that Hsp90 and mutated KIT were valid molecular targets in the therapy of AML.