Additional file 1 of A novel semiautomatic Chinese keywords instrument screening delirium based on electronic medical records

Additional file 1

Comprehensive profiling of the metabolome in corn silage inoculated with or without <i>Lactiplantibacillus plantarum</i> using different untargeted metabolomics analyses

Silage fermentation is a complicated biochemical process involving interactions between microbes and metabolites. However, the overall metabolome feature of ensiled forage and its response to lactic acid bacteria inoculation is poorly understood. Hence, in this study metabolome profiles of whole-plant corn silage inoculated with or without Lactiplantibacillus plantarum were characterised via solid-phase microextraction/gas chromatography/mass spectrometry (SPME-GC-MS), gas chromatography/time-of-flight mass spectrometry (GC-TOF-MS), and Liquid chromatography/Q Exactive HFX mass spectrometry (LC-QE-MS/MS) analysis. There were 2087 identified metabolites including 1143 reliably identified metabolites in fresh and ensiled whole-plant corn. After ensiling, the increased metabolites in whole-plant corn were mainly composed of organic acids, volatile organic compounds (VOC), benzene and substituted derivatives, carboxylic acids and derivatives, fatty acyls, flavonoids, indoles and derivatives, organooxygen compounds (including amines and amides), phenols, pyridines and derivatives, and steroids and steroid derivatives, which includes neurotransmitters and metabolites with aromatic, antioxidant, anti-inflammatory, and antimicrobial activities. Phenylacetaldehyde was the most abundant aromatic metabolite after ensiling. L-isoleucine and oxoproline were the major free amino acids in silage. Ensiling markedly increased the relative abundances of 3-phenyllactic acid, chrysoeriol, 6-O-acetylaustroinulin, acetylcholine, γ-aminobutyric acid, pyridoxine, and alpha-linoleic acid. Inoculation with L. plantarum remarkably changed silage VOC composition, and essential amino acids, 3-phenyllactic acid, and cinnamaldehyde compared with untreated silage. The present study does not only provide a deeper insight into metabolites of the ensiled whole-plant corn but also reveals metabolites with specific biological functions that could be much helpful in screening novel lactic acid bacteria to well ensile forages. Inoculation with L. plantarum significantly affects the metabolome in ensiled whole-plant corn.</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>Concentration and purity of total RNA isolated from leaves of <i>Salvia miltiorrhiza</i> and <i>Platycodon grandiflorus</i> using 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 <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

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

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

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