New bisesquiterpenoid lactone from the wild rhizome of <i>Atractylodes macrocephala</i> Koidz grown in Qimen

<p>The rhizomes of <i>Atractylodes macrocephala</i> are used as both a food source and traditional Chinese medicine in China. A phytochemical investigation was carried out on wild <i>A. macrocephala</i> grown in Qimen County in eastern China, and yielded a novel bisesquiterpenoid lactone, namely, biatractylenolide II (<b>1</b>), along with two known compounds, atractylenolide II (<b>2</b>) and taraxeryl acetate (<b>3</b>). The structure and relative configuration of the new compound were elucidated mainly by 1D and 2D NMR spectroscopic methods in combination with HRESIMS experiments. This paper describes the isolation and structural elucidation of the new bisesquiterpenoid lactone (<b>1</b>).</p

Comprehensive analysis of complete chloroplast genome sequence of <i>Plantago asiatica</i> L. (Plantaginaceae)

Plantago asiatica L. is a representative individual species of Plantaginaceae, whose high reputation is owed to its edible and medicinal values. However, the phylogeny and genes of the P. asiatica chloroplast have not yet been well described. Here we report the findings of a comprehensive analysis of the P. asiatica chloroplast genome. The P. asiatica chloroplast genome is 164,992 bp, circular, and has a GC content of 37.98%. The circular genome contains 141 genes, including 8 rRNAs, 38 tRNAs, and 95 protein-coding genes. Seventy-two simple sequence repeats are detected. Comparative chloroplast genome analysis of six related species suggests that a higher similarity exists in the coding region than the non-coding region, and differences in the degree of preservation is smaller between P. asiatica and Plantago depressa than among others. Our phylogenetic analysis illustrates P. asiatica has a relatively close relationship with P. depressa, which was also divided into different clades with Plantago ovata and Plantago lagopus in the genus Plantago. This analysis of the P. asiatica chloroplast genome contributes to an improved deeply understanding of the evolutionary relationships among Plantaginaceae.</p

Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using the modified CHAN method.

<p>Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using the modified CHAN method.</p

A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using <i>Dendrobium huoshanense</i>

<div><p>Acquiring high quality RNA is the basis of plant molecular biology research, plant genetics and other physiological investigations. At present, a large number of nucleotide isolation methods have been exploited or modified, such as commercial kits, CTAB, SDS methods and so on. Due to the nature of different plants, extraction methods vary. Moreover, efficiency of certain approach cannot be guaranteed due to composition of different plants and extracting high quality RNA from plants rich in polysaccharides and polyphenols are often difficult. The physical and chemical properties of polysaccharides which are similar to nucleic acids and other secondary metabolites will be coprecipitated with RNA irreversibly. Therefore, how to remove polysaccharides and other secondary metabolites during RNA extraction is the primary challenge. <i>Dendrobium huoshanense</i> is an Orchidaceae perennial herb that is rich in polysaccharides and other secondary metabolites. By using <i>D</i>. <i>huoshanense</i> as the subject, we improved the method originated from CHAN and made it suitable for plants containing high amount of polysaccharides and polyphenols. The extracted total RNA was clear and non-dispersive, with good integrity and no obvious contamination with DNA and other impurities. And it was also evaluated by gel electrophoresis, nucleic acid quantitative detector and PCR assessment. Thus, as a simple approach, it is suitable and efficient in RNA isolation for plants rich in polysaccharides and polyphenols.</p></div

Concentration and purity of total RNA isolated from leaves of <i>Salvia miltiorrhiza</i> and <i>Platycodon grandiflorus</i> using modified CHAN method.

<p>Concentration and purity of total RNA isolated from leaves of <i>Salvia miltiorrhiza</i> and <i>Platycodon grandiflorus</i> using modified CHAN method.</p

Agarose gel electrophoresis of the RT-PCR tubulin products.

<p>Lane 1, lane 2 and lane 3 showed RT-PCR products from <i>D</i>. <i>huoshanense</i> stem, leaf and flower. Lanes 0 showed RT-PCR product by substituting <i>D</i>. <i>huoshanense</i> cDNA for ddH₂O. Marker: DL2,000 DNA marker.</p

Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using original CHAN method.

<p>Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using original CHAN method.</p

Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using Trizol method.

<p>Concentration and purity of total RNA isolated from <i>D</i>.<i>huoshanense</i> stem, leaf and flower using Trizol method.</p

1.0% agarose gel electrophoresis of total RNA isolated.

<p>A, three intact RNA bands for 28S, 18S and 5S RNA. Lane 1, lane 2 and lane 3 in A, B, C, D and E contain 1 μg of total RNA from <i>D</i>. <i>huoshanense</i> stem, leaf and flower, respectively. A: modified CHAN method; B: original CHAN method; C: Trizol method; D: RNeasy Plant Mini Kit method; E: RNAprep Pure Plant Kit method.</p

DataSheet1_Paeoniflorin alleviates 17α-ethinylestradiol-induced cholestasis via the farnesoid X receptor-mediated bile acid homeostasis signaling pathway in rats.docx

Cholestasis, characterized by disturbance of bile formation, is a common pathological condition that can induce several serious liver diseases. As a kind of trigger, estrogen-induced cholestasis belongs to drug-induced cholestasis. Paeoniflorin is the most abundant bioactive constituent in Paeonia lactiflora Pall., Paeonia suffruticosa Andr., or Paeonia veitchii Lynch, a widely used herbal medicine for treating hepatic disease over centuries in China. However, the pharmacologic effect and mechanism of paeoniflorin on estrogen-induced cholestasis remain unclear. In this experiment, the pharmacological effect of paeoniflorin on EE-induced cholestasis in rats was evaluated comprehensively for the first time. Ultra-high-performance liquid chromatography coupled with Q-Exactive orbitrap mass spectrometer was used to monitor the variation of bile acid levels and composition. It was demonstrated that paeoniflorin alleviated 17α-ethinylestradiol (EE)-induced cholestasis dose-dependently, characterized by a decrease of serum biochemical indexes, recovery of bile flow, amelioration of hepatic and ileal histopathology, and reduction of oxidative stress. In addition, paeoniflorin intervention restored EE-disrupted bile acid homeostasis in enterohepatic circulation. Further mechanism studies using western blot, quantitative Real-Time PCR, and immunohistochemical showed that paeoniflorin could upregulate hepatic efflux transporters expression but downregulate hepatic uptake transporter expression. Meanwhile, paeoniflorin reduced bile acids synthesis by repressing cholesterol 7α-hydroxylase in hepatocytes. Paeoniflorin affected the above transporters and enzyme via activation of a nuclear receptor, farnesoid X receptor (FXR), which was recognized as a vital regulator for maintaining bile acid homeostasis. In conclusion, paeoniflorin alleviated EE-induced cholestasis and maintained bile acid homeostasis via FXR-mediated regulation of bile acids transporters and synthesis enzyme. The findings indicated that paeoniflorin might exert a potential therapeutic medicine for estrogen-induced cholestasis.</p