Alangium chinense is a commonly used medicinal plant of Alangiaceae Alangium, one of the characteristic Miao medicines in Guizhou. The genus is strongly differentiated and has complex morphological variation, with significant differences in active ingredients and potency among herbs. In this paper, we sequenced the chloroplast genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib using Illumina high-throughput sequencing technology. The genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplasts were sequenced, their assembly annotation and structural characterization were completed, and the genomes of the congeners Alangium chinense and Alangium alpinum were downloaded from NCBI for comparative analysis. Finally, the phylogenetic tree was constructed by selecting published species with close affinity to Alangium chinense family from NCBI. The results were as follows: Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplast genomes were 156 670, 156 672 and 156 871 bp in length, with a typical four-segment structure, all containing one long single copy region (LSC), one short single copy region (SSC) and two inverted repeat regions (IRa and IRb). All of them were annotated to 133 genes, including 88 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Three types of long repeat sequences and SSR loci, forward repeats, palindrome repeats and reverse repeats, were found in all chloroplast genomes. Comparative genomic analysis showed that there was diversity in the boundary transition regions of the five species, no rearrangements or inversions were found in the chloroplast genome, and there were significant differences in the coding regions of ndhA, ycf1, rpl16, ycf2, and petD genes, which provided new loci for molecular identification of Patagonia. In the phylogenetic analysis, Alangium kurzii var. kurzii clustered as a single species, and Alangium chinense subsp. Pauciflorum and Alangium chinense subsp. Strigosum clustered as a single species, with a support rate of 93 and close affinity, indicating that the phylogenetic tree can be used for the species identification. This paper can provide a basis for the taxonomic identification of Alangium, ensure the safety of clinical use of anise herbs and regulate the market of Alangium chinense herbs.

For rapid discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (M^pro) inhibitors from a natural product library, a novel colorimetric screening assay was developed. According to the colorimetric principle, the synthetic peptide TSAVLQ-para-nitroanilide (pNA) was used as the M^pro hydrolysis substrate. Subsequently, the working concentration of pNA substrate, M^pro working concentration, hydrolysis time and DMSO tolerance were optimized for the development of a simple and robust colorimetric screening assay. Through these systematic optimizations, we selected 0.4 μmol·L^-1 M^pro and 100 μmol·L^-1 pNA substrate as the optimal working concentrations in this colorimetric screening assay, and a high Z' factor of 0.9 was achieved. Using this screening assay, natural product ginkgolic acid C13:0 (GA13:0) was identified as a novel competitive M^pro inhibitor in vitro. Taken together, we have successfully developed a simple and optimized colorimetric screening assay, which will be vital for the discovery of novel SARS-CoV-2 M^pro inhibitors.

The enteric-hepatic axis plays an important role in the occurrence, progression and regression of nonalcoholic fatty liver disease (NAFLD). Obeticholic acid (OCA) is a farnesoid X receptor agonist. In this study, C57BL/6 mice were fed with methionine and choline deficient (MCD) diet for 8 weeks with OCA (6.5 mg·kg^-1·d^-1) administration by gavage at the same time. The effects of OCA on serum lipid and bile acid metabolomics and ileal gut microbiota (GM) of MCD mice were studied by UPLC-MS and 16S rDNA sequencing. The results were as follows: (1) OCA decreased the activities of alanine aminotransferase and aspartate aminotransferase in serum and the contents of triglyceride (TG) and malondialdehyde in liver, alleviated the accumulation of liver fat and inflammation of MCD mice. OCA down-regulated the contents of 2 eicosanoids (12, 13-EPOME, 9, 10-EPOME) and 4 free fatty acids (FFA16∶1, FFA18∶1, FFA16∶2, FFA18∶3) and TG (16∶1_16∶1_18∶2) in serum, and up-regulated the content of 1 eicosanoid thromboxanes B3. KEGG differential metabolite pathway analysis showed that fatty acid biosynthesis might be the main way that OCA ameliorated lipid metabolism disorder of MCD mice. OCA reduced the relative abundance of Christensenellaceae and Lachnospiraceae_UCG-006 in the GM of MCD mice; OCA decreased the serum levels of 23-deoxycholic acid, porcine deoxycholic acid, 3-deoxycholic acid, glycine deoxycholic acid, glycine cholic acid, taurine deoxycholic acid, taurocholic acid and taurine. These results suggest that the alleviating effect of OCA on NAFLD of MCD mice may be related to its above-mentioned regulation of the metabolism of the free fatty acids, oxidized lipids, 12α-hydroxylated bile acids and the abundance of GM. The animal experiments were approved by the Experimental Animal Ethics Committee of Hubei University (No. 20220036).

A new [7, 10∶1, 5]-patchoulane-type sesquiterpenoid was isolated and purified by silica gel column chromatography, medium pressure liquid chromatography, and semi-preparative high performance liquid chromatography. Based on IR, HR-ESI-MS, NMR, and X-single crystal diffraction data analyses, compound 1 was determined to be (-)-(3S, 4R, 5R, 7R, 10R)-[7, 10∶1, 5]patchoul-1(2)-en-3, 4-diol. It showed an inhibitory effect on LPS-induced NO production in RAW264.7 cells.

Heterotrimeric G proteins are classes of signal-transducing proteins that bind to guanine nucleotides and possess GTP hydrolase activity. G proteins are composed of three subunits α, β, and γ, and are considered as the "molecular switch" in the GPCR signaling pathway. The abnormal activation of G protein is strongly related to diseases such as uveal melanoma, asthma, et al., making directly targeting G protein as an promising strategy for combating diseases. In this review, the classification and physiological functions of G protein are briefly described, and the research progress of G proteins in diseases, G protein modulators are reviewed.

In this study, we investigated the effect of aspirin on tumor biological effects mediated by hepatocyte growth factor/cellular-mesenchymal-epithelial transition factor (HGF/c-Met) axis, and preliminarily explored the molecular mechanism of inhibiting tumor metastasis by aspirin. The binding of aspirin to c-Met was predicted by molecular docking; cellular thermal shift assay (CETSA) was used to verify the binding of aspirin to c-Met at the cellular level. The inhibitory effect of aspirin on c-Met kinase was detected by kinase activity; Western blot, cell scattering test, cell branching morphogenesis and Transwell test were used to evaluate the cell signal transduction, morphological changes and migration and invasion ability. The results showed that aspirin could effectively inhibit the kinase activity of c-Met with a half inhibitory concentration of 0.95 mmol·L^-1. The results of docking showed that aspirin could bind to the ATP pocket of c-Met protein, and the main binding sites were Tyr1230, Tyr1159 and Met1229. The CETSA test also showed that aspirin could form binding complex with c-Met protein. Western blot results showed that aspirin could inhibit the up-regulation of phosphorylated Met stimulated by HGF in a concentration-dependent manner. The results of cell scattering test showed that aspirin could block HGF/c-Met promoted cell scattering in a concentration dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. Similarly, the results of MDCK cell branching morphogenesis experiment showed that aspirin could inhibit HGF/c-Met mediated invasive growth in a concentration dependent manner. The results of Transwell test showed that aspirin could block HGF/c-Met mediated cell migration and invasion in a concentration-dependent manner. Aspirin could almost completely block the biological function mediated by c-Met activation at the concentration of 4 mmol·L^-1, and this effect was independent of HGF. The above results indicate that aspirin can bind to c-Met, thereby blocking the biological effects mediated by HGF/c-Met, and inhibiting tumor metastasis. This study revealed the new biological function of aspirin, and provided a new theoretical basis for a comprehensive understanding of the anti-metastatic effect of aspirin.

Model-informed drug development (MIDD) in the development of pediatric drugs is drawing more and more attention due to the insufficiency of subjects, lack of research on ontogeny, and the limitation of ethic. The core of MIDD used for dose selection includes the population pharmacokinetic (PopPK) model and physiologically based pharmacokinetic (PBPK) model, as well as model-based simulation and prediction. PBPK model has the advantage of predicting the optimal pediatric dose before the clinical trials and has the ability of extrapolation from adult model to pediatric model. PopPK model characterizes the pediatric PK feature based on the analysis of clinical data and can be used to explore the significant covariates, which is a power tool for individualized medicine in children. With their own advantages and disadvantages, PBPK and PopPK model should be jointly used in the pediatric drug development to refine the dose regimen for children at different ages. In this study, the pediatric drug development of rivaroxaban was taken as an example to introduce the combined application of PBPK model and PopPK model in the design and validation of pediatric dose regimen in Phase Ⅰ, Ⅱ and Ⅲ trials, which may provide reference to MIDD in other pediatric drug development.

In this study, a research strategy integrating network pharmacology analysis and animal experimental validation was applied to explore the molecular mechanism of Chuanxiong Qingnao Granules (CXQN) in improving migraine headache (MH). All animal experiments were followed the regulation of the Laboratory Animal Ethics Committee of the China Academy of Chinese Medical Sciences. Based on the network pharmacology analysis, the 27 active ingredients and their corresponding 940 targets were obtained, and 99 common targets of CXQN in the treatment of MH were obtained by intersection, and tumor necrosis factor-α (TNF-α), interleukin (IL)-6, vascular endothelial growth factor A (VEGFA), IL-1β, brain-derived neurotrophic factor (BDNF) were screened out as hub targets. Enrichment analysis showed that the targets of CXQN in the treatment of MH were mainly involved in cyclic adenosine monophosphate (cAMP), hypoxia inducible factor-1 (HIF-1), phosphoinositide 3-kinase-protein kinase B (PI3K-Akt) signaling pathways. In addition, the experimental verification in the MH rat induced by nitroglycerin showed that the CXQN administrated groups could significantly improve the behavioral symptoms and regulate the level of vasoactive substances, and reduce the expression of TNF-α, IL-6, VEGFA, IL-1β, and BDNF at gene and protein levels. This study revealed the multi-component, multi-target, and multi-pathway characteristics of CXQN in the treatment of MH, and elucidated the potential mechanism of CXQN in the treatment of MH, laying a theoretical foundation and scientific basis for its clinical application in the treatment of MH diseases.

Nine compounds were isolated from the leaves of Chimonanthus nitens Oliv. by silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. They were identified as chimnitensene B (1), 1α-hydroxyisodauc-4-en-15-al (2), trans-4, 5-dihydroxycorocalane (3), trefoliol B (4), oplopanone (5), oplodiol (6), 3-(3'-hydroxybutyl)-2, 4, 4-trimethylcyclohexa-2, 5-dienone (7), 9(S)-4-oxo-7, 8-dihydro-β-ionol (8), and saniculamoid D (9) respectively, by MS, NMR and single crystal diffraction. Among them, compound 1 is a new guaiane-sesquiterpenoid, and compounds 2-9 were isolated from this plant for the first time.

The urate transporter 1 (URAT1) which controls urate reabsorption is a membrane transporter in the apical membrane of human renal proximal tubule epithelial cells. It was found that about 90% of patients suffer from hyperuricemia due to insufficient uric acid excretion. Therefore, the development of URAT1 inhibitors that can reduce the level of serum uric acid in vivo by enhancing renal urate excretion has been a hot spot in seeking anti-gout drugs in recent years. In this article, the representative URAT1 inhibitors with uric acid-lowering or anti-gout effects are reviewed, and related medicinal chemical strategies are analyzed, hoping to provide valuable insights into the discovery of new URAT1 inhibitors.