Wei Jia

This study investigated the preservative effect and molecular mechanism of cold-air pre-cooling (CAP) combined with storage at 15 °C/85% relative humidity on Volvariella volvacea. CAP significantly reduced weight loss and browning, maintained moderate respiratory intensity, minimised malondialdehyde accumulation and polyphenol oxidase activity, and preserved higher firmness and soluble-protein content, extending the shelf life by 4 d. An analysis of energy metabolism indices revealed that CAP increased mitochondrial quantity, membrane potential, and ATP content. Specifically, CAP restricted the tricarboxylic acid (TCA) cycle rate by downregulating the activities of succinate dehydrogenase, isocitrate dehydrogenase, and citrate synthase. Additionally, CAP prevented the peak activation of respiratory complex I, while sustaining optimal activity of complexes III and IV, thereby stabilising intracellular ATP levels. Transcriptomic analysis further indicated that CAP suppressed the activity of the TCA cycle and oxidative phosphorylation pathways during postharvest storage. Quantitative real-time PCR (qPCR) validated the downregulation of genes associated with respiratory complexes after CAP treatment. In conclusion, CAP maintained the postharvest quality of V. volvacea by preserving energy metabolism homeostasis, providing a theoretical basis for its application in edible mushroom preservation.

Lyophyllum decastes soluble dietary fiber (LDSDF) is a polysaccharide-based active ingredient derived from the edible and medicinal fungus L. decastes. However, its extraction methods remain unoptimized, and its hypolipidemic and gut microbiota effects have yet to be thoroughly investigated in mice. In this study, response surface optimization of the LDSDF extraction method indicated an optimal extraction temperature of 99 °C, a solid/liquid ratio of 25:1 mL/g, and an extraction time of 1.9 h. The optimal ethanol precipitation parameters were a concentration ratio of 3.9, an ethanol concentration of 74.4%, and a precipitation time of 16.4 h. These conditions afforded an LDSDF yield of 15.83%. Following 6 weeks of oral gavage of LDSDF in obese mice, the results showed that LDSDF inhibited increases in body and organ weight; reduced serum levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol; increased serum levels of high-density lipoprotein cholesterol; decreased alanine aminotransferase and aspartate aminotransferase activities; and lowered systemic levels of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6, and interleukin-1β). Concurrently, it elevated the hepatic activities of superoxide dismutase, catalase, and glutathione peroxidase; reduced malondialdehyde levels; and mitigated lesions in liver and epididymal fat cells. Meanwhile, 16S rRNA sequencing revealed that LDSDF significantly alleviated intestinal flora imbalances. Overall, this study established an optimized extraction process to obtain LDSDF with a high yield and confirmed the hypolipidemic and gut microbiota-modulating efficacy of this active ingredient, highlighting its potential for use as a functional food ingredient.

FVPB1, a novel heteropolysaccharide, was extracted from the Flammulina velutipes fruiting body, and its structure was determined by methylation analysis, nuclear magnetic resonance (NMR) spectroscopy. FVPB1 demonstrated efficacy in inhibiting lipid accumulation in Raw264.7 cells and zebrafish, as well as in reducing weight gain and ameliorating liver injury in high-fat diet-induced mice. High concentration of FVPB1 significantly increased serum ApoA1 levels, while all tested doses (low, medium, and high) reduced serum ApoB levels in mice. Intervention with FVPB1 significantly increased the mRNA expression of Lcat and Cyp7a1 enzymes while markedly reducing the transcriptional level of Hmgcr reductase. Additionally, low concentration of FVPB1 enhanced CYP7A1 protein expression, whereas medium and high concentrations of FVPB1 promoted LCAT protein expression. Medium and high concentrations of FVPB1 significantly enhanced bile acid excretion in mice, with the high dose additionally promoting fecal sterol output. Alpha and beta diversity analyses demonstrated that a high-fat diet induced substantial dysbiosis in the gut microbiota of mice, characterized by reduced microbial diversity and richness. Intervention with FVPB1 significantly modulated the structural composition of the intestinal microbiota in high-fat diet-fed mice. Therefore, FVPB1 exerts lipid-lowering effect in high-fat diet-fed mice by modulating cholesterol metabolism and ameliorating gut microbiota dysbiosis.

The structure and characteristics of LEOPs, β-oligosaccharides from the fruiting body of Lentinus edodes obtained via acid degradation and gel permeation chromatography, were investigated. We performed high-performance liquid chromatography, infrared spectroscopy, methylation analysis, nuclear magnetic resonance, and correlated activity experiments, including antioxidant, immunomodulatory, and liver injury protection to gain insights. LEOPs comprised an oligosaccharide (Mw 2445 Da) based on six β-1, 3-D-glucose residues as the main chain and six β-1, 6-D-glucose residues as the side chain. Surface plasmon resonance analysis indicated that LEOPs directly bound to dectin-1, which facilitated their immunoenhancing activity via downstream NF-κB activation. The results implied that LEOPs may be the active unit of the shiitake β-glucan. The determination of LEOPs structure was performed to reveal the anti-tumor effect and immune-regulatory function of shiitake β-glucan on a molecular level to provide a basis.

FVPT1, a novel heteropolysaccharide, was purified from the fruiting body of Flammulina velutipes using magnetic-field-assisted three-phase partitioning and gel permeation chromatography. The structure was characterized using monosaccharide composition and methylation analysis, infrared spectroscopy and nuclear magnetic resonance (NMR). The FVPT1 (~1.64 × 10⁴ Da) was composed of L-fucose, D-galactose, D-glucose and D-mannose at a molar ratio of 1.0:3.5:1.0:1.4. The polysaccharide repeating unit of FVPT1 was established with methylation analyses and NMR spectroscopy. Moreover, a zebrafish larva hyperlipidemia model test demonstrated that FVPT1 can show appreciable lipid-lowering effects. In addition, the FVPT1 exhibited remarkable immunoregulatory activity by increasing nitric oxide, interleukin (IL)-1β and IL-1 secretion in macrophages. Therefore, these results suggest that FVPT1 has the potential to be developed into a new immune or hypolipidemic health product.

Antrodin C, a bioactive component of Taiwanofungus camphoratus, exhibits good immunophysiological and antitumour activities, including a broad spectrum of anticancer effects. Exogenous additives can bind to metabolites during the submerged culture of T. camphoratus and affect secondary metabolite yields. However, the lack of molecular genetic studies on T. camphoratus has hindered the study of the antrodin C biosynthetic pathway. In this study, we conducted a ribonucleic acid-sequencing-based transcriptional analysis to identify the differentially expressed genes involved in the synthesis of antrodin C by T. camphoratus, using inositol and maleic acid (MAC) as exogenous additives. The addition of inositol significantly upregulated carbohydrate and sugar metabolism pathway genes (E3.2.1.14, UGDH, and IVD). When MAC was used, amino and nucleotide sugar metabolism and starch and sucrose metabolism pathways were significantly inhibited, and the associated genes (E3.2.1.14 and E3.2.1.58) were also significantly downregulated. The biosynthesis pathway genes for ubiquinone and other terpene quinones (COQ2, ARO8, and wrbA), which may play an important role in antrodin C synthesis, were significantly downregulated. This study advances our understanding of how the additives inositol and MAC affect metabolite biosynthesis in T. camphorates. This could be beneficial in proposing potential strategies for improving antrodin C production using a genetic approach.

The current study aims to explore the possible anti-lung carcinoma activity of ADC as well as the underlying mechanisms by which ADC exerts its actions in NSCLC. Findings showed that ADC potently inhibited the viability of SPCA-1, induced apoptosis triggered by ROS, and arrested the cell cycle at the G2/M phase via a P53 signaling pathway. Interestingly, phenomena such as autophagosomes accumulation, conversion of the LC3-I to LC3-II, etc., indicated that autophagy could be activated by ADC. The blockage of autophagy-augmented ADC induced inhibition of cell proliferation, while autophagy activation restored cell death, indicating that autophagy had a protective effect against cell death which was induced by ADC treatment. Meanwhile, ADC treatment suppressed both the Akt/mTOR and AMPK signaling pathways. The joint action of both ADC and the autophagy inhibitor significantly increased the death of SPCA-1. An in vitro phase I metabolic stability assay showed that ADC was highly metabolized in SD rat liver microsomes and moderately metabolized in human liver microsomes, which will assist in predicting the outcomes of clinical pharmacokinetics and toxicity studies. These findings imply that blocking the Akt/mTOR signaling pathway, which was independent of AMPK inhibition, could activate ADC-induced protective autophagy in non-small-cell lung cancer cells.

Polysaccharides from the immunomodulatory medicinal mushroom Inonotus obliquus (IOPS) were extracted and purified using three-phase partitioning (TPP), which is an efficient, fast, safe, and green purification technique. An optimal extraction procedure that gave a good 2.2% isolated yield was identified, using the following protocol: a solid-liquid ratio of 1 g to 12 mL; mass fraction of (NH₄)₂SO₄ 20% (w/v); 11 mL t-butanol; pH 8.0; temperature 30 °C; and extraction time 30 min. The purified IOPS was shown to be a proteoglycan of 40 kDa molecular weight comprising of d-galactose, d-glucose, d-xylose, and d-mannose in a molar ratio of 2.0:3.5:1.0:1.5. The purified IOPS displayed strong free-radical scavenging abilities, antioxidant activities, and immunological activity in vitro. IOPS’ Trolox antioxidant equivalent capacity and ferric-reducing ability of plasma were 251.2 μmol Trolox/g sample and 1040.5 μmol Fe²⁺/g sample, respectively, with the activity of its immunomodulatory behavior shown to be gradient dependent.

Taiwanofungus camphoratus has been reported to have antitumor effects against various cancer cells. The aim of this study was to investigate the direct inhibitory effect of By-1 (3-isobutyl-l-methoxy-4-[4'-(3-methylbut-2-enyloxy)phenyl]-1H-pyrrole-2,5-dione), a compound from spent broth from submerged cultures of T. camphoratus, on human lung adenocarcinoma cells and to determine the molecular mechanism underlying this effect. The growth-inhibitory assay and colony formation assay showed that cell viability was significantly decreased. A By-1 concentration of 300 μmol/L caused 73.55% cell death and at a concentration of 240 μmol/L led to a 58% reduction in the number of colonies. The wound-healing assay showed that the distance of migration was 0.3 times shorter than that of untreated cells. Flow cytometry revealed that By-1 could suppress DNA synthesis, cause cell cycle arrest at the S phase, and induce apoptosis in a reactive oxygen species-dependent manner. Furthermore, the expression of caspase-3 and P53 was 4 times higher than that in untreated cells, and the antiapoptotic protein Bcl-2 was decreased 2 times compared with the protein in untreated cells. It is interesting to note that apoptosis and autophagy were both induced during treatment with By-1, and autophagy inhibition decreased cell proliferation. By-1 potently inhibited the growth of SPCA-1 cells by inducing cell cycle arrest and apoptosis. The combination of proapoptosis agents and antiautophagy agents could effectively enhance anticancer efficacy, which may be a new strategy in treating non-small cell lung cancer.