Lipidomic and transcriptomic profiles of glycerophospholipid metabolism during Hemerocallis citrina Baroni flowering.

BACKGROUND: Hemerocallis citrina Baroni (daylily) is a horticultural ornamental plant and vegetable with various applications as a raw material in traditional Chinese medicine and as a flavouring agent. Daylily contains many functional substances and is rich in lecithin, which is mostly composed of glycerophospholipids. To study the comprehensive dynamic changes in glycerophospholipid during daylily flowering and the underlying signalling mechanisms, we performed comprehensive, time-resolved lipidomic and transcriptomic analyses of 'Datong Huanghua 6' daylily. RESULTS: Labelling with PKH67 fluorescent antibodies clearly and effectively helped visualise lipid changes in daylily, while relative conductivity and malonaldehyde content detection revealed that the early stages of flowering were controllable processes; however, differences became non-significant after 18 h, indicating cellular damage. In addition, phospholipase D (PLD) and lipoxygenase (LOX) activities increased throughout the flowering process, suggesting that lipid hydrolysis and oxidation had intensified. Lipidomics identified 558 lipids that changed during flowering, with the most different lipids found 12 h before and 12 h after flowering. Transcriptome analysis identified 13 key functional genes and enzymes in the glycerophospholipid metabolic pathway. The two-way orthogonal partial least squares analysis showed that diacylglycerol diphosphate phosphatase correlated strongly and positively with phosphatidic acid (PA)(22:0/18:2), PA(34:2), PA(34:4), and diacylglycerol(18:2/21:0) but negatively with phospholipase C. In addition, ethanolamine phosphotransferase gene and phospholipid-N-methyltransferase gene correlated positively with phosphatidylethanolamine (PE)(16:0/18:2), PE(16:0/18:3), PE(33:2), and lysophosphatidylcholine (16:0) but negatively with PE(34:1). CONCLUSIONS: Overall, this study elucidated changes in the glycerophospholipid metabolism pathway during the daylily flowering process, as well as characteristic genes, thus providing a basis for future studies of glycerophospholipids and signal transduction in daylilies.

[Screening and confirmation of pesticide residues in substitutional tea such as orange peel, lotus leaf, pueraria lobata and Pangdahai].

OBJECTIVE: The screening method was established for pesticide residues in substitutional teas such as orange peel, lotus leaf, pueraria lobata and Pangdahai using data base. The method for analyzing of 26 pesticide residues was confirmed in four substitutional teas with gas chromatography-tandem mass spectrometry(GC-MS/MS) and dispersive solid-phase extraction(d-SPE) for sample preparation. METHODS: The 26 types of pesticides were selected as target compounds, including screened out, commonly used in substitutional tea planting and forbidden in agriculture planting. The samples extracted with acetonitrile, and purified by adsorbent(e. g PSA, GCB, C_(18)), the purification solutions were separated on DB-5 MS UI column((15 m+PUU+15 m)×0. 25 mm×0. 25 µm) with programmed temperature and determined by GC-MS/MS in multi-reaction monitoring(MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity(r≥0. 995)in concentration range(2-100 µg/L) for 26 kinds of pesticides. The limits of quantification(LOQ) were 3-40 µg/kg. The average recoveries range were 71. 9%-114. 4% in three add levels of 10, 50, 200 µg/kg, and relative standard deviations of 1. 0%-16. 0%(n=6). In the four matrixes, the pesticides were detected concentrating on orange peel, and there were kinds of residues. CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of pesticide screening and simultaneous analysis of multiple pesticide residues in orange peel, lotus leaf, pueraria lobata and Pangdahai.

[Determination of pesticide residues in substitutional tea by dispersive solid-phase extraction and gas chromatography-triple quadrupole mass spectrometry].

OBJECTIVE: The method was established for determination 14 kinds of psticide residues in substitutional tea using gas chromatography-triple quadrupole mass spectrometry( GC-MS/MS) for instrument method and dispersive solid-phase extraction( dSPE) for sample preparation. METHODS: The samples extracted with acetonitrile, and purified by N-propyl ethlene diamine( PSA), the extracts were separated on DB-5MS column( 30 m × 0. 25 mm, 0. 25 µm) with programmed temperature and determined by GCMS/MS in multi-reaction monitoring( MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity( r ≥0. 999) in certain ranges for 14 kinds of pesticides. The limits of quantification( LOQ)were 4-40 µg/kg. The average recoveries range were 65. 6%-116. 6% in three add levels of 20, 40 and 200 µg/kg, and relative standard deviations( RSD S) of 1. 5%-15. 7%( n = 6). CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of simultaneous analysis of multiple pesticide residues in substitutional tea.

Ameliorative effects of Qingfei Tongluo formula on experimental mycoplasmal pneumonia in mice.

Mycoplasma pneumoniae pneumonia (MPP) is a common disease in children. Qingfei Tongluo formula (QTF) has been used for the treatment of MPP clinically, but the chemical constituents and mechanism involved remain unclear. This study aimed to analyze the main chemical constituents and to explore the possible mechanism of action associated with QTF treatment of MPP. Liquid chromatography-mass spectrometry was employed to identify the compounds contained in the QTF extract. A BALB/c mouse model of MP infection was established. After treatment with QTF (0.85 and 1.70 g/kg) for 3 days, hematoxylin and eosin staining was performed in lung tissues for histological examination. Inflammatory cytokines were detected by ELISA. Western blot analysis was used for detecting phosphorylated proteins involved in MAPK and nuclear factor (NF)-κB signaling pathways. In the mouse model, a large amount of pulmonary interstitial infiltration of lymphocytes and plasmacytes were seen as well as bronchus and vasodilation congestion. Following QTF treatment, inflammation was alleviated significantly compared with the model group. Inflammatory cytokines [interleukin (IL)-6, transforming growth factor-ß1, IL-8, IL-1ß and tumor necrosis factor-α] in bronchoalveolar lavage fluid were decreased dramatically. In addition, we found that QTF inhibited activation of phosphorylation of JNK, ERK and NF-κB. In conclusion, QTF alleviates MPP inflammation possibly via inhibitory activation of MAPK/NF-κB pathways, which can act as a new agent for MPP treatment.