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A novel water-soluble polysaccharide (HEP-4) with a molecular weight of 1.98 × 105Da was extracted from honeysuckle. Structural characterization was performed using high-performance liquid chromatography (HPLC), gas chromatography, Fourier transform-infrared (FT-IR) spectrum, nucleus magnetic resonance (NMR) spectra, and scanning electron microscopy. The results showed that HEP-4 is primarily composed of mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose with a mole ratio of 6.74:1.56:1.04:14.21:4.31:5.4, and the major types of the glycosidic bond types of HEP-4 were 1-α-D-Glcp, 1,4-β-D-Glcp, 1-β-D-Arap, 1,3,4-β-D-Arap, and 1,3,6-β-D-Manp. The results of bioactivity experiments revealed that HEP-4 had antioxidant in vitro. In addition, HEP-4 inhibited H2O2-induced oxidative damage and increased the activity of HepG2 cells by reducing MDA levels and inhibiting ROS production. Meanwhile, HEP-4 significantly enhanced the activities of GSH-Px and CAT, indicating that HEP-4 exerts a protective effect on H2O2-induced oxidative stress. These results indicate that HEP-4 could be a potential natural antioxidant.

The decreased number of viable bacteria and the ability of Bifidobacterium to adhere to and colonize the gut in the gastrointestinal environment greatly limit their efficacy. To solve this problem, thiolated carboxymethyl cellulose sodium (CMC) probiotic double-layered multinucleated microcapsules with Bifidobacterium adolescentis FS2-3 in the inner layer and Bacillus subtilis SN15-2 embedded in the outer layers were designed. First, the viable counts and release rates of microcapsules were examined by in vitro simulated digestion assays, and it was found that microcapsules were better protected from gastrointestinal digestion than the controls. Compared with free Bifidobacterium strains, double-layered multinucleated microcapsules have higher viable bacterial survival rates and storage stability. Second, through in vitro rheology, tensile tests, isotherm titration calorimetry, and adhesion tests, it was observed that thiolated CMC could enhance the strong interaction of Bifidobacterium with intestinal mucus and significantly promote the proliferation and growth of probiotics. Finally, double-layered multinucleated microcapsules containing B. adolescentis FS2-3 and B. subtilis SN15-2 modified with sulfhydryl-modified CMC were studied in the intestine. Alleviation of Escherichia coli O157:H7 induced intestinal inflammation. The results showed that microencapsulation could significantly increase the colon content of Bifidobacterium, relieve intestinal inflammation symptoms in mice with bacterial enteritis, and repair the intestinal microbiota disorder caused by inflammation. The probiotic double-layered multinucleated microcapsules prepared in this study can improve the survival rate of probiotics and promote proliferation, adhesion, and colonization of probiotics.

Aim: This study aimed to develop and validate an efficient LC–MS/MS method for the simultaneous determination of Hcy, Cys, Met and 5-methyltetrahydrofolate in human serum and to apply this method to patients with coronary artery disease. Methodology and results: Serum samples were prepared by reduction with dithiothreitol followed by protein precipitation. The analytical run time was 2.2 min. The linearity was good in the range of 2–100 μmol/l-1 for Hcy and Met, 10–500 μmol/l-1 for Cys, and 1–50 ng/ml-1 for 5-methyltetrahydrofolate. Conclusion: An accurate and precise method that was rapid, robust and with high-throughput for the routine clinical monitoring of patients with coronary artery disease was developed.