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Orthogonal experiment was applied to optimize the water extraction parameters of zinc from Flammulina velutipes, and then the extracts were separated by membrane filter (0.45 mm) and D101 macroporous resin. Six different species of Zn were obtained and the Zn content of various species were determined by flame atomic absorption spectrometry. The optimized conditions for the extraction of Zn were: ratio of dried material to water, 1:30; extraction temperature, 75°C; extraction time, 120 minutes. About 34.43 mg Zn was extracted from 1 g dried F. velutipes powder under the optimal conditions. The recovery value for Zn was 96.5% with a low relative standard deviation. In addition, the content of the organic state of Zn was more than that of the inorganic state, and most of the organic state Zn was found in the polysaccharide and protein fractions. [ABSTRACT FROM AUTHOR]

Overactivation of Wnt/b-catenin pathway due to dysfunction of retinoid-related orphan receptor a (RORa) is related to cancer development and progression. Diallyl disulfide (DADS), an active component of garlic, has been reported in our previous study for upregulation of RORa expression in gastric cancer (GC) cells. It remains to be elucidated the role and mechanism of RORa in DADS against GC. This study revealed that DADS treatment resulted in reduced expression levels of Wnt1, b-catenin, TCF-4, intranuclear b-catenin and p-b-catenin in GC cells, concomitant with the compromised expression of b-catenin target genes (Axin, c-Jun, and c-Myc). RORa overexpression augmented DADS-induced downregulation of Wnt1/b-catenin pathway, G2/M phase arrest, and cell growth inhibition in vitro and in vivo. Contrarily, knockdown of RORa attenuated these effects of DADS. Interestingly, DADS induced an increase in the binding of RORa to b-catenin, which may lead to reduction of b-catenin phosphorylation and nuclear translocation. This interplay modulated by DADS may affect b-catenin target gene expression for that the opposite results were observed in DADS-treated RORa knockdown and overexpression cells. DADS caused a decrease in vimentin, snail and MMP-9, as well as an increase in E-cadherin and TIMP3 expression, which restricted epithelialemesenchymal transition (EMT), migration, and invasion. The aforementioned effects of DADS were weakened simultaneously when the suppression of DADS on the Wnt1/b-catenin pathway was resisted by knockdown of RORa. In contrast, overexpression of RORa enhanced the effects of DADS. Therefore, RORa-mediated downregulation of Wnt1/b-catenin pathway could undertake an important role in anticancer activity of DADS against GC cell proliferation, EMT, migration, and invasion. [ABSTRACT FROM AUTHOR]