Characterization and identification of chemical constituents in Corni Fructus and effect of storage using UHPLC-LTQ-Orbitrap-MS.

Corni Fructus (CF) has been widely used as both traditional medicine and food; however, systematic studies on its chemical profile and the impact of storage periods on the indicative components are lacking. In this study, UHPLC-LTQ-Orbitrap-MS was used to investigate the fragmentation behaviors of multiple compounds from CF and the content variety of its indicative components for different storage periods. The major basic components of CF were determined to be iridoid glucosides, pentacyclic triterpenoids, phenolic acids, tannins and flavonoids. The characteristic cleavage pathways of the iridoid glucosides, pentacyclic triterpenoids, phenolic acids, tannins and flavonoids were further investigated and elaborated, which could assist in identifying the structures of similar components of other Chinese herbal medicines. Using accurate mass measurements for each precursor ion and the subsequent fragmented ions, and then comparing with standards and literature data, a total of 130 components, including 69 iridoid glucosides, 9 pentacyclic triterpenoids, 16 phenolic acids, 20 tannins and 16 flavonoids, 47 of which are potentially new compounds, were identified. The storage period studies indicated that the contents of 19 indicative components in CF changed differently with the prolongation of the storage period. Among them, morroniside, loganin, sweroside, cornuside, gallic acid, oleanolic acid and ursolic acid were the most important. These results provide abundant information for the identification and improved understanding of the chemical constituents in CF to clarify the content variety of its indicative components for different storage periods.

Tea consumption and risk of lower respiratory tract infections: a two-sample mendelian randomization study.

BACKGROUND: Observational studies have reported the association between tea consumption and the risk of lower respiratory tract infections (LRTIs). However, a consensus has yet to be reached, and whether the observed association is driven by confounding factors or reverse causality remains unclear. METHOD: A two-sample Mendelian randomization (MR) analysis was conducted to determine whether genetically predicted tea intake is causally associated with the risk of common LRTI subtypes. Genome-wide association study (GWAS) from UK Biobank was used to identify single-nucleotide polymorphisms (SNPs) associated with an extra cup of tea intake each day. The summary statistics for acute bronchitis, acute bronchiolitis, bronchiectasis, pneumonia, and influenza and pneumonia were derived from the FinnGen project. RESULTS: We found that genetically predicted an extra daily cup of tea intake was causally associated with the decreased risk of bronchiectasis [odds ratio (OR) = 0.61, 95% confidence interval (CI) = 0.47-0.78, P < 0.001], pneumonia (OR = 0.90, 95% CI = 0.85-0.96, P = 0.002), influenza and pneumonia (OR = 0.91, 95% CI = 0.85-0.97, P = 0.002), but not with acute bronchitis (OR = 0.91, 95% CI = 0.82-1.01, P = 0.067) and acute bronchiolitis (OR = 0.79, 95% CI = 0.60-1.05, P = 0.100). Sensitivity analyses showed that no heterogeneity and pleiotropy could bias the results. CONCLUSIONS: Our findings provided new evidence that genetically predicted an extra daily cup of tea intake may causally associated with a decreased risk of bronchiectasis, pneumonia, and influenza and pneumonia.

Six undescribed derivatives of stilbene isolated from Lindera reflexa Hemsl. and their anti-tumor and anti-inflammatory activities.

Six undescribed stilbene derivatives Reflexanbene DH (1-4, 6) and Reflexanbene J (5), as well as one known stilbene 3,5-dimethoxystilbene (7), were isolated from the dried roots of Lindera reflexa Hemsl. Their structures and absolute configurations were elucidated using spectroscopy and electronic circular dichroism (ECD) analysis. In cytotoxic assays, moderately inhibitory activities of Reflexanbene F (3) against MGC80-3 and A549 cell lines were observed, with IC50 values of 15.42 and 5.09 µM, respectively. The IC50 value of Reflexanbene E (2) on A549 cell lines was 19.78 µM. The isolated compounds were also tested for their inhibitory effect against LPS-induced NO and IL-6 production in RAW 264.7 cells. In particular, Reflexanbene J (5) and Reflexanbene H (6) showed significant inhibition of NO production in LPS-stimulated macrophage RAW 264.7 cells at the concentration of 20 µM. Furthermore, the expression of IL-6 protein in the LPS-induced RAW 264.7 cells can also be significantly inhibited by different concentrations (5, 10 and 20 µM, p < 0.05 or p < 0.01) of compounds 1-7.

Common mineral nutrients in ankylosing spondylitis: A 2-sample Mendelian randomization study.

OBJECTIVE: The causal relationship between common mineral nutrients and ankylosing spondylitis (AS) has not been studied. So this Mendelian randomization (MR) study aims to investigate the causal association of varying levels of calcium, zinc, copper, and selenium on AS. DESIGN: We selected 4 elements potentially associated with the onset and development of AS as exposure factors, single nucleotide polymorphisms (SNPs) as instrumental variables, and these SNPs are independent of each other(r2 < 0.05) and highly correlated with each of the 4 elements (P < 5 × 10-8 ). The 2-sample MR method takes Inverse-variance weighted (IVW) and MR-Egger as the main method and Simple mode (SM), Weighted median (WM1 ), and Weighted mode (WM2 ) as supplementary methods to evaluate the causal effect of mineral levels on AS. RESULTS: The IVW analysis does not provide convincing evidence to support a causal association between calcium (odds ratio [OR] = 1.000, 95% CI = 0.994, 1.005, P = .875), copper (OR = 1.000, 95% CI = 1.000, 1.001, P = .533) and selenium (OR = 0.999, 95% CI = 0.998, 1.000, P = .229) and AS. The IVW (OR = 1.001, 95% CI = 1.000, 1.002, P = .029) and WM1 (OR = 1.001, 95% CI = 1.000, 1.002, P = .011) results of zinc show that per standard deviation increment in zinc is a suggestive association with risks of AS, and MR-Egger (OR = 1.004, 95% CI = 0.996, 1.013, P = .265) and other supplementary methods indicate that zinc is not causally associated with AS. All MR-Egger intercept parameters and MR Pleiotropy RESidual Sum and Outlier tests demonstrated the absence of horizontal pleiotropy. CONCLUSIONS: This study does not provide convincing evidence to support a causal correlation between calcium, zinc, copper, and selenium with AS.

Changes in the Chemical Components of Processed Rehmanniae Radix Distillate during Different Steaming Times.

Distillate was obtained in different processing cycles of processed Rehmanniae Radix (PRR). In this study, we investigated the chemical compositions of distillates 1 (Dis1) to 9 (Dis9) via GC-MS and LC-MS. Differences between Dis1-Dis9 were noticeable. A total of 13 and 21 compounds were detected via GC-MS and LC-MS, respectively, including organic acids, furans, alcohols, iridoid glycosides, phenylpropanoid glycosides, and saccharides. The relative contents of compound 2,5-hydroxymethylfurfural and furans all gradually increased with steaming time. Other compounds, however, exhibited a negative trend or fluctuated. Of these compounds, iridoid glycosides and phenylpropanoid glycosides were unstable and easily degraded, which led to a gradually decreasing concentration with increased steaming times. In addition, the degradation products were mainly derived from catalpol and acteoside, among which catalpol mainly existed as aglycone and its rearranged products. However, acteoside was converted into verbasoside through the removal of caffeoyl. Some volatile alcohols, such as phenylethyl alcohol, hydroxyphenyl ethanol, and 3-hydroxy-4-methoxybenzoic acid, were also likely from the degradation of acteoside and its homologs. These results provide an important reference basis for the processing methods, quality evaluation, and rational clinical application of PRR and its distillate.

Comprehensive 16S rDNA Sequencing and LC-MS/MS-Based Metabolomics to Investigate Intestinal Flora and Metabolic Profiles of the Serum, Hypothalamus and Hippocampus in p­Chlorophenylalanine-Induced Insomnia Rats Treated with Lilium brownie.

Gut microbiota homeostasis in the organism and insomnia have been reported to influence each other. In the study, a method of 16S rRNA gene sequencing combined with ultra-high performance liquid chromatography-mass/mass spectrometry was adopted to evaluate the effects of Lilium brownie (LB) on intestinal flora and metabolic profiles of serum, hypothalamus and hippocampus in insomnia rat induced by p­chlorophenylalanine (PCPA). It was observed that the imbalance in the diversity and abundance of gut microbiota induced by PCPA was restored after LB intervention. Among these, the Porphyromonadaceae, Lactobacillus and Escherichia were significantly adjusted at the genus level by PCPA and LB, respectively. It was also found that the most of metabolic phenotypes in serum, hypothalamus and hippocampus perturbed by PCPA were regulated towards normal after LB intervention, especially 5-hydroxy-L-tryptophan of the hypothalamus involving in 5-HT metabolism. Moreover, the arachidonic acid metabolism in serum, hypothalamus and hippocampus, and the serotonergic synapse in hypothalamus and hippocampus were the most fundamentally and significantly affected pathways after LB intervention. The results of correlation analysis showed that several floras including Pseudoruegeria have an outstanding contribution to the change of differential metabolites. In brief, the results confirm that gut microbiota is significantly returned to normal and may interact with the corresponding metabolites to relieve insomnia under LB intervention.

A Universal and Reversible Wet Adhesive via Straightforward Aqueous Self-Assembly of Polyethylenimine and Polyoxometalate.

The excellent adhesion of mussels under wet conditions has inspired the development of numerous catechol-based wet adhesives. Nevertheless, the performance of catechol-based wet adhesive suffers from the sensitivity toward temperature, pH, or oxidation stimuli. Therefore, it is of great significance to develop non-catechol-based wet adhesives to fully recapitulate nature's dynamic function. Herein, a novel type of non-catechol-based wet adhesive is reported, which is readily formed by self-assembly of commercially available branched polyethylenimine and phosphotungstic acid in aqueous solution through the combination of electrostatic interaction and hydrogen bonding. This wet adhesive shows reversible, tunable, and strong adhesion on diverse substrates and further exhibits high efficacy in promoting biological wound healing. During the healing of the wound, the as-prepared wet adhesive also possesses inherent antimicrobial properties, thus avoiding inflammations and infections due to microorganism accumulation.

Chemical constituents of the antiulcer purified fractions of Lindera reflexa Hemsl. and its quantitative analysis.

The root of Lindera reflexa Hemsl. (LR) is a folk Chinses herbal medicine that has been used to treat gastritis and peptic ulcers. In this study, three new stilbenes (1-3) and two known flavonoids (4 and 5) were isolated from the antiulcer purified fractions of LR. The chemical structures of the isolated compounds were characterized comprehensively based on the basis of extensive spectroscopic data. Absolute configurations of compounds 1, 2, and 3 were determined by ECD calculations. The cytotoxic activities of compounds 1-5 were evaluated by MTT assay. Compound 4 showed the strongest inhibitory effect on the proliferation of tumor cells lines MGC803 and SMMC-7721, with IC50 values of 2.65 and 4.13 µM, respectively. The quantitative analysis of 12 compounds of the antiulcer purified fractions of LR were carried out by using the reversed-phase high-performance liquid chromatography (HPLC) method. Within the test range, all calibration curves showed good linearity (R2 > 0.9993). The LOD, LOQ, specificity, precision, and accuracy of the method were verified. Therefore, the present study may provide a valuable method for quality control the antiulcer purified fractions of LR.

Screening, and identification of the binding position, of xanthine oxidase inhibitors in the roots of Lindera reflexa Hemsl using ultrafiltration LC-MS combined with enzyme blocking.

A method based on enzyme blocking combined with ultrafiltration liquid chromatography-mass spectrometry (LC-MS) has been developed to identify xanthine oxidase (XOD) inhibitors in the roots of Lindera reflexa Hemsl (LR) and determine their binding positions. Allopurinol and febuxostat, known XOD inhibitors, which occupy different binding positions in XOD, were used as blockers and pre-incubated with XOD. Then the LR extract was incubated without XOD, and with XOD, allopurinol-blocked XOD and febuxostat-blocked XOD before ultrafiltration LC-MS was performed. By comparing the chromatographic profiles of the incubation samples, not only the ligands, but also the binding position of these ligands with XOD could be determined. Finally, three compounds, pinosylvin, pinocembrin and methoxy-5-hydroxy-trans-stilbene, were identified as potential XOD inhibitors and the binding modes of these three compounds were shown to be similar to those of febuxostat. To verify the XOD inhibitory activity of the screened compounds, the microplate method and molecular docking in silico were used to evaluate the enzyme inhibitory activities and the binding positions with XOD. The results showed that the developed method could screen for XOD ligands in LR extracts and also determine the binding positions of the ligands. To our knowledge, this is the first report of the XOD inhibitory activity of these three compounds.

Pharmacokinetics and Tissue Distribution Study of Pinosylvin in Rats by Ultra-High-Performance Liquid Chromatography Coupled with Linear Trap Quadrupole Orbitrap Mass Spectrometry.

Pinosylvin is a potential anti-inflammatory and antioxidant compound and the major effective medicinal ingredient in the root of Lindera reflexa Hemsl. However, few investigations have been conducted regarding the pharmacokinetics, excretion, characteristics of tissue distribution, and major metabolites of pinosylvin in rats after oral administration. To better understand the behavior and mechanisms of action underlying the activity of pinosylvin in vivo, we established a simple, sensitive, and reliable ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for quantifying pinosylvin in rat plasma, urine, feces, and various tissues (including heart, liver, spleen, lung, kidneys, large intestine, small intestine, and stomach). Noncompartmental pharmacokinetic parameters indicated that pinosylvin is rapidly distributed and taken up by tissues. The time to peak (maximum) concentration (Tmax) was 0.137 h, and the apparent elimination half-life (t1/2) was 1.347±0.01 h. The results of the tissue distribution study suggest that pinosylvin is widely distributed to various tissues; the highest concentration was observed after 10 min in the stomach, followed by the heart, lung, spleen, and kidneys. Results of the excretion study suggest that a small amount of pinosylvin is excreted from the urine and feces in the parent form; the 73 h accumulative excretion ratios of urine and feces were 0.82% and 0.11%, respectively. It is likely that pinosylvin is mostly metabolized in vivo. Nine metabolites were found, and the main metabolic pathways of pinosylvin in rats included glucuronidation, hydroxylation, and methylation. Four metabolites had higher concentrations in the stomach, suggesting that the stomach is a potential target organ of pinosylvin. In conclusion, the present study may provide a material basis for studying the pharmacological action of pinosylvin and provides meaningful information for the clinical treatment of chronic gastritis and gastric ulcers using Radix Linderae Reflexae.

Characterization and identification of the chemical constituents in the root of Lindera reflexa Hemsl. using ultra-high performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry.

The root of Lindera reflexa Hemsl. (LR) is a newly discovered herbal drug and has been used to treat gastritis and peptic ulcers. Nevertheless, the chemical profile of LR has not been established. In this study, ultra-high performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap-MS) was performed to investigate the fragmentation behaviors of multiple compounds from LR. The 12 standards were divided into five types according to their basic skeletons: stilbenes, flavonoids, alkaloids, pyranone I and pyranone II. The MS(n) spectra of the [M+H](+) or [M±NH4](+) ions for these compounds provided a wealth of structural information on the five different types of compounds. Stilbenes yielded ions with successive loss of 78Da (C6H6) and 28Da (CO). The subsequent loss of H2O, CO, RDA and C-ring fragmentation were the most possible fragmentation pathways for flavonoids. Fragmentation with successive loss of 17Da (NH3) or 31Da (CH5N), 32Da (CH4O) and 28Da (CO) in the MS(n) spectra were characteristics of alkaloids. The characteristic ions for Pyranone I were m/z 255.1013, m/z 243.1013 and m/z 237.0909, and the diagnostic ions for Pyranone II were m/z 227.0700, m/z 215.0700, m/z 185.0594 and m/z 131.0489. Using accurate mass measurements for each precursor ion and the subsequent fragmented ions, a total of 42 compounds were identified or tentatively characterized in LR, including 24 potentially new compounds.

Isolation and cytotoxic activity of selaginellin derivatives and biflavonoids from Selaginella tamariscina.

Five selaginellin derivatives, including two new selaginellins termed selaginellins M (1) and N (2), and three previously identified compounds, selaginellin (3), selaginellin A (4), and selaginellin C (5), were isolated from the Selaginella tamariscina (Beauv.) Spring plant. In addition, four known biflavonoids, namely neocryptomerin ( 6), hinokiflavone (7), pulvinatabiflavone (8), and 7''- O-methylamentoflavone (9), were also isolated. The structures of new compounds 1 and 2 were elucidated by spectroscopic analysis. The cytotoxic activity of compounds 1- 9 was evaluated against a small panel of human cancer cell lines, including U251 (human glioma cells), HeLa (human cervical carcinoma cells), and MCF-7 (human breast cancer cells). The two new selaginellins, selaginellins M (1) and N (2), showed medium activity against the human cancer cell lines.