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Objective We aimed to (i) investigate the protective effect of active components of water extract of huangqi (milkvetch root) on high glucose (25 mmol/L) induced injury of human umbilical vein endothelial cells (HUVEC) and (ii) elucidate the underlying mechanism. Methods Male SD rats were continuously fed with water extract [20 g/(kg·d)] for 7 d, and blood samples were colleeted 1, 2 and 4 h after the last gavage, combined with supernatant, and performed ultra-performance liquid chromatography (UPLC-MS) analysis to screen the active components of the extract in rats. Cells were divided into normal group, model group, different active components groups treated with different concentrations, and the concerted application of active components group. Cell viability was measured by Cell Counting Kit-8 to explore the optimal concentration of the effective components of milkvetch root. Oxidative damage was measured by reactive oxygen species (ROS). The enzyme-linked immunosorbent assay measured the contents of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA); the protein expression level of nuclear factor E2 related factor (Nrf 2) and heme oxygenase 1 (HO-1) by Western blotting. Results Formononetin, daidzein, and calycosin are the active components of milkvetch root water extract.Compared with the normal group, the cell viability of the model group was increased (P<0.05). Compared with the model group, cell viability was decreased in each concentration(26, 13, 6.5 μmol/L) of formononetin group (P<0.05), a high dose of daidzein (0.11 μmol/L) could reduce cell viability (P<0.05). Compared with the normal group, the ROS levels in the model group were increased(P<0.05); compared with the model group, the ROS levels in the formononetin high and medium concentrations(26, 13 μmol/L) groups, different concentrations of daidzein (0.11, 0.06, 0.03 μmol/L), the calycosin high concentration group (7.00 μmol/L) were decreased (P<0.05). The combined active components inhibitive rate was higher than the single active components, and highest in the high concentration group. Compared with the normal group, Nrf2 and HO-1 protein expressions were decreased in the model group (P<0.05); compared with the model group, Nrf2 and HO-1 proteins were increased in the group with high concentration of concerted application group (P<0.05). Compared with the normal group, SOD, CAT were decreased (P<0.05), MDA increased (P<0.05) in the model group; compared with the model group, SOD, CAT in the high concentration concerted application group were increased (P<0.05), MDA was significantly decreased (P<0.05). Conclusion The active components of milkvetch root water extract, formononetin, daidzein, calycosin can protect HUVEC from high glucose-induced injury. The mechanism may be related to the regulation of Nrf2, HO-1, SOD, CAT, and MDA expression and their antioxidant effects. [ABSTRACT FROM AUTHOR]

In order to develop a novel effective biological insecticide for controlling oleander hawk moth, a new pathogen was isolated from naturally diseased Daphnis nerii. Based on scanning electron microscopy, full-length amplification of cDNAs (FLAC), and phylogenetic analysis of genome segments 2and 10,the virus was identified as a new type of cypovirus (Da phnis nerii cypovirus [DnCPV]). Electrophoresis analysis showed that DnCPV had a genome comprising 10double-stranded RNA (dsRNA) segments, ranging from 892 to 4160bp.Using FLAC, the cDNAs from the 10 dsRNA segments of the new CPV were cloned and genome segments 2and 10 were sequenced. Sequencing results showed that segment 2 encoded RNA-dependent-RNA-polymerases (RdRps) and segment 10 encoded polyhedrin. These two segments shared conserved terminal sequences of AGUCAAA and AGC at the 5’and 3’ends,respectively.These conserved terminal sequences were not consistent with any of the known CPV types.Phylogenetic analysis of the RdRp and polyhedrin indicated that this CPV was more closely related to CPV type 19 and type 5than other CPV types. Based on the unique conserved terminal sequences and the electrophoresis pattern of the new virus, we tentatively named it DnCPV Nanchang isolate: DnCPV-NC.