Diterpenoid alkaloids from the root of <i>Aconitum sinchiangense</i> W. T. Wang with their antitumor and antibacterial activities

<p>A phytochemical study of the root barks of <i>Aconitum sinchiangense</i> W. T. Wang, a traditional Chinese herb medicine, led to the isolation of 15 diterpenoid alkaloids, including one new C₁₉-diterpenoid alkaloid, sinchiangensine A (<b>1</b>), whose structure was determined by spectral methods including 2D NMR. Additionally, sinchiangensine A and its known analogue <b>3</b> were first reported as potential antitumor and antibacterial diterpenoid alkaloids, which showed significant antitumor activities against tumour cells (HL-60, A-549, SMCC-7721, MCF-7 and SW480), with IC₅₀ comparable to cisplatin, and significant antibacterial activities against <i>Staphylococcus aureus</i> ATCC-25923 with MIC value of 0.147 and 0.144 μmol/mL, respectively.</p

Additional file 2: Figure S2. of Duck enteritis virus (DEV) UL54 protein, a novel partner, interacts with DEV UL24 protein

WB analyzed the expression of UL24-fusion protein and UL54-fusion protein in HEK293T cells. HEK293T cells were transfected with eukaryotic plasmid pCMV-myc, pCMV-myc-UL24, and pCMV-Flag-UL54 respectively. At 48 h post-infection, the 293 T cell extracts were carried out Western blotting analysis, which indicated that myc-UL24 and Flag-UL54 was expressed in 293 T cells and the molecular mass of fusion protein is about 45 KD, 50.5 KD respectively. Primary Abs against myc-UL24 and Flag-UL54 were serums of rabbit against UL24 and mouse against Flag respectively. (PDF 44 kb

MOESM1 of Microbial community compositions in the gastrointestinal tract of Chinese Mongolian sheep using Illumina MiSeq sequencing revealed high microbial diversity

Additional file 1. Additional tables and figures

Influence of different treatment conditions of resveratrol on DEV infection.

<p>DEFs infected with DEV (MOI  = 0.1) were treated with resveratrol in various mode of actions. (1) Virus inactivation: DEV was treated with drug for 1 h at 37°C before infection. The virus and drug mixture was then added to cells for 1 h at 37°C. The media was removed and replaced with drug-free media. DEV suspended without drug was used as control (effect of temperature). (2) Inhibition of virus attachment: DEFs were infected in media containing the drug and virus for 1 h at 37°C, and then the mixture was removed. The cells were overlaid with drug-free media. (3) Pre-treatment effect: DEFs were pre-treatment with drug for 1 h at 37°C. And then DEV was added to the cells after resveratrol solution removal. After 1 h 37°C, the inoculum was removed and replaced with drug-free media. (4) Intracellular inhibition: after removing the unabsorbed virus, the media containing the resveratrol was added to the DEFs. At 48 h p.i., the total DNA was extracted from DEV-infected DEFs and copies of DEV were evaluated by the real-time FQ-PCR assay. Values are means ± SD (n = 3). Significance: different capital letters represent extremely significant differences among groups (P<0.01, n = 3).</p

Standard curve of real-time FQ-PCR amplification.

<p>Serial 10-fold dilutions of standard DEV DNA from 5.25×10¹⁰ to 10² copies were amplified in this process. Amplification efficiency (E) was 99.8%. The standard curve equations was <i>C</i>t = −3.328×lg [virus copies/5.25]+40.138 (R² = 0.997).</p

Inhibition effects of resveratrol on DEV <i>in vitro</i>^a.

a<p>The inhibition effects on DEV were evaluated by MTT assay.</p>b<p>Inhibition concentration 50% (IC₅₀): concentration required to inhibit DEV at 72 h post-infection by 50%.(n = 3).</p>c<p>Cytotoxic concentration 50% (CC₅₀) concentration required to reduce cell viability by 50%. (n = 3).</p>d<p>SI: Selectivity index is defined as the radio of CC₅₀ to IC₅₀.</p

Growth curves of DEV in DEFs in the presence or absence of reveratrol.

<p>The DEFs were firstly infected with DEV at MOI of 1 and test resveratrol solution for 1 h at 37°C. DEV proliferated in the DEFs in the presence or absence of reveratrol within 72 h. The total DNA was extracted at the indicated time points (2, 4, 6, 8, 10, 12, 24, 48, and 72 h p.i.) and copies of DEV were detected via real-time FQ-PCR assay. Copies of DEV were calculated according to the standard curve equations: <i>C</i>t = −3.328×lg [virus copies/5.25]+40.138 (R² = 0.997). Values are means ± SD (n = 3).</p

Additional file 2: of Virulent duck enteritis virus infected DEF cells generate a unique pattern of viral microRNAs and a novel set of host microRNAs

Table S2. Distribution of sRNAs in DEV-infected and uninfected samples. (DOCX 15 kb

Additional File 10:

Figure S3. Regulatory network of DEF miRNAs and CHv genes. a Gene regulatory network formed by differentially-expressed DEF miRNAs (blue ellipses) and their target genes (yellow rectangles). b Gene regulatory network of differentially-expressed DEF miRNAs (blue circles) and target immediate-early (IE) genes (yellow rectangles). (PDF 353 kb

Additional file 9: of Virulent duck enteritis virus infected DEF cells generate a unique pattern of viral microRNAs and a novel set of host microRNAs

Table S7. Differentially expressed host alignment miRNA in CHv-infected DEF cells. Fold-change = log2 (infected/mock in expression) > 1 or < − 1, and p-value < 0.05 indicated significance differentially expressed miRNA. Fold-change < − 1 indicates down-regulated, Fold-change > 1 indicates up-regulated. (XLSX 12 kb