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ABSTRACTRaddeanin A (RA) has indicated suppressive effects on various human tumor cells, and insufficient vitamin D was associated with human papillomavirus (HPV) persistence and gynecological tumors. However, combined effects of RA and vitamin D on HPV-positive cells remain elusive. Herein, we aimed to investigate the combined effects of RA and 1ɑ,25(OH)2D3(VD3) on cellular viability and modulation of HPV18E6/E7, programmed cell death 1 ligand (PD-L1) and vitamin D receptor (VDR) expression in HeLa cells in vitro. HeLa cells were treated with RA alone or VD3 combined with RA. Cell viability was measured using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), and apoptosis was detected by flow cytometry. Real-time PCR (qRT-PCR) and Western blot were used to determine the gene/protein expression levels. The autophagosomes were observed by Transmission electron microscopy (TEM). The result showed that cell viability was inhibited by RA, and apoptosis in HeLa cells treated with RA was elevated accordingly. The expression of Bax, Cleaved-caspase-3, Cleaved-caspase-9 and Cleaved-PARP increased, and Bcl-2 decreased. The autophagy was induced by RA, as evidenced by elevated autophagosomes and the increased LC3-II/I ratio and Beclin-1. The expression of HPV18E6/E7, PD-L1 and VDR was reduced by RA. Moreover, RA combined with VD3 had a stronger effect on HeLa cells than RA alone. In conclusion, RA inhibits HeLa proliferation and induces apoptosis and autophagy via suppressing HPV18E6/E7, PD-L1 and VDR, and VD3 showed reinforced effects of RA on HeLa cells. Therefore, combined usage of VD3 with RA might be a potential novel immunotherapy strategy for HPV-related diseases.

AbstractFlavonoids are important polyphenols, and widely distributed in plants. Anthocyanidin reductase (ANR), a precursor enzyme in (-)- epicatechin synthesis, could play an important role in anthocyanins biosynthesis. In the present study, the ANRgene was cloned from Vitis viniferaL. cv.Cabernet Sauvignon. Compared with VvANR (GenBank Accession No. NM_001280956.1), they differed in only 4 bp and shared similarity of 99.6%. The ANR protein belongs to the cl33424 Superfamily. In addition, many light-responsive elements and abiotic stress-related elements (ABRE, MYB, WRKY, etc.) were found in the promoter region of the ANRgene. STRING analysis predicted that VvMYBPA1and VvMYBPARmay be involved in regulating the expression of the ANRgene. Different UV-C irradiation times were used for Cabernet Sauvignon berries during grape berry development, and the comparison analysis showed that UV-C irradiation improved significantly the accumulation of catechin and epicatechin during 20–50 days after full bloom, especially, under UV-C irradiation for 5 min. Furthermore, UV-C irradiation for 5 min had a greater effect on the expression of the ANRgene, the accumulation of catechin and epicatechin, and the enzymatic activity of ANR than UV-C irradiation for 10 min. Through the virus-induced gene silencing system, the ANR-silenced grape leaves were chlorotic, in which the contents of catechin and epicatechin were both significantly lower than those in the control, showing a very significant difference, respectively. In summary, our results demonstrated that the ANRgene played a crucial role in the accumulation of epicatechin in grapes, and its expression was upregulated by UV-C irradiation.

This study investigated the chemical compositions of essential oils (EOs) from ten taxa belonging to the Pinus genus. The studied taxa that grow wildly in China, and the EOs were extracted by steam distillation; the chemical compositions were isolated and characterized by GC-MS. Eighty-one components, representing over 92.10% of the EOs, were identified. The main constituents of EOs were α-pinene (6.44%–53.00%), β-caryophyllene (2.43%–24.64%), β-pinene (0.00%–22.32%), δ-cadinene (2.56%–17.56%), germacrene D (0.74%–11.38%), and camphene (0.78%–10.48%). Furthermore, we determined the total phenolic content (TPC) of the EOs, with the values from 26.50 to 60.01 mg eq GAE/mL EO. DPPH free radical scavenging activity (DPPH), ferric reducing antioxidant power (FRAP), and ABTS radical cation scavenging activity (ABTS) were used to evaluate the antioxidant capacity of the EOs, and the obtained values were ranged from 499.15 to 1,272.75 mmol eq Trolox/mL EO, 1,255.67 to 3,857.93 mmol eq Trolox/mL EO, and 370.81 to 1,677.19 mmol eq Trolox/mL EO, respectively. The results showed that all of the EOs studied had strong antioxidant activity, and these pine plants could be used as natural antioxidants in functional food and pharmaceutical industries.