SLC25A32 promotes malignant progression of glioblastoma by activating PI3K-AKT signaling pathway.

BACKGROUND: Solute carrier family 25 member 32 (SLC25A32) is an important member of SLC25A family and plays a role in folate transport metabolism. However, the mechanism and function of SLC25A32 in the progression of human glioblastoma (GBM) remain unclear. METHODS: In this study, folate related gene analysis was performed to explore gene expression profiles in low-grade glioma (LGG) and GBM. Western blotting, real-time quantitative PCR (qRT-PCR), and immunohistochemistry (IHC) were used to confirm the expression levels of SLC25A32 in GBM tissues and cell lines. CCK-8 assays, colony formation assays, and Edu assays were performed to assess the role of SLC25A32 on proliferation in GBM in vitro. A 3D sphere invasion assay and an ex vivo co-culture invasion model were performed to assess the effects of SLC25A32 on invasion in GBM. RESULTS: Elevated expression of SLC25A32 was observed in GBM, and high SLC25A32 expression was associated with a high glioma grade and poorer prognosis. Immunohistochemistry performed with anti-SLC25A32 on samples from an independent cohort of patients confirmed these results. Knockdown of SLC25A32 inhibited the proliferation and invasion of GBM cells, but overexpression of SLC25A32 significantly promoted cell growth and invasion. These effects were mainly due to the activation of the PI3K-AKT-mTOR signaling pathway. CONCLUSION: Our study demonstrated that SLC25A32 plays a significant role in promoting the malignant phenotype of GBM. Therefore, SLC25A32 can be used as an independent prognostic factor in patients with GBM, providing a new target for the comprehensive treatment of GBM.

Changes of Bioactive Components and Antioxidant Capacity of Pear Ferment in Simulated Gastrointestinal Digestion In Vitro.

Fruit ferment is rich in polyphenols, organic acids, enzymes, and other bioactive components, which contribute to their antioxidant ability. In this study, we investigated the effect of the simulated gastric and intestinal digestion in vitro on the total phenolic content (TPC), total flavonoid content (TFC), phenolic components content, organic acid content, protease activity, superoxide dismutase (SOD) activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (DPPH-RSA), hydroxyl (·OH) radical scavenging activity (·OH-RSA), and total reducing capacity in 'Xuehua' pear (Pyrus bretschneideri Rehd) ferment. The result showed that the TPC, TFC, protease activity, and phenolic components such as arbutin, protocatechuic acid, malic acid, and acetic acid showed a rising trend during the simulated gastric digestion in 'Xuehua' pear ferment, and these components might contribute to the increasing of ·OH-RSA and total reducing capacity. The SOD activity and epicatechin content showed an increasing trend at first and then a decreasing trend, which was likely associated with DPPH-RSA. During in vitro-simulated intestinal digestion, the majority of evaluated items reduced, except for protease activity, quercetin, and tartaric acid. The reason for the decreasing of bio-accessibility resulted from the inhibition of the digestive environment, and the transformation between substances, such as the conversion of hyperoside to quercetin. The correlation analysis indicated that the antioxidant capacity of 'Xuehua' pear ferment was mainly affected by its bioactive compounds and enzymes activity as well as the food matrices and digestive environment. The comparison between the digestive group with and without enzymes suggested that the simulated gastrointestinal digestion could boost the release and delay the degradation of phenolic components, flavonoids, and organic acid, protect protease and SOD activity, and stabilize DPPH-RSA, ·OH-RSA, and total reducing capacity in 'Xuehua' pear ferment; thus, the 'Xuehua' pear ferment could be considered as an easily digestible food.

Shigella flexneri T3SS effector IpaH4.5 modulates the host inflammatory response via interaction with NF-κB p65 protein.

Shigella species possess a type III secretion system (T3SS), which is required for human infection and that delivers effector proteins into target host cells. Here, we show that the effector, IpaH4.5 dampens the pro-inflammatory cytokine response. In both the Sereny test and a murine lung infection model, the Shigella ΔipaH4.5 mutant strain caused more severe inflammatory responses and significantly induced higher pro-inflammatory cytokine levels (MIP-2 and TNF-α) in the lung homogenates of wild type-infected mice. Moreover, there was a threefold decrease in bacterial colonization of the mutant compared with the WT and ΔipaH4.5/ipaH4.5-rescued strains. Yeast two-hybrid screening showed that IpaH4.5 specifically interacts with the p65 subunit of NF-κB. Ten truncated versions of IpaH4.5 and p65 spanning different regions were constructed and expressed to further map the IpaH binding sites with p65. The results revealed thatthe p65 region spanning amino acids 1-190 of p65 interacted with the IpaH4.5/1-293 N-terminal region. In vitro, IpaH4.5 displayed ubiquitin ligase activity towards ubiquitin and p65. Furthermore, the transcriptional activity of NF-κB was shown to be inhibited by IpaH4.5 utilizing a dual-luciferase reporter gene detection system containing NF-κB promoter response elements. Thus, we conclude that the IpaH4.5 protein is an E3 ubiquitin ligase capable of directly regulating the host inflammatory response by inhibiting the NF-κB signalling pathway.

Aqueous two-phase extraction, identification and antioxidant activity of anthocyanins from mulberry (Morus atropurpurea Roxb.).

Aqueous two-phase extraction (ATPE), identification and antioxidant activity of anthocyanins from mulberry (Morus atropurpurea Roxb.) were investigated in this study. The optimal differential partitioning of mulberry anthocyanins (MAY) and sugars was achieved in a system (pH 4.5, temperature=35±1°C) composed of 30% (w/w) ethanol, 20% (w/w) concentration of ammonium sulphate, 10% (w/w) mulberry juice and 40% (w/w) water. The multiple partitioning indicated a single ATPE step could isolate the majority of anthocyanins, while removing near to 90% of the free sugars. The composition of the MAY extracted by ATPE had no obvious changes, and five anthocyanins were identified by HPLC-ESI-MS/MS. The ATPE extract showed a relatively high antioxidant ability compared to that by the conventional extraction. Our results indicated that ATPE was a valuable method for the removal of the majority of free sugars, which held great promise in the purification of natural anthocyanin pigments.