This study was designed to investigate the role of activation of the ferroptosis pathway in the inhibition of esophageal cancer proliferation and metastasis by realgar, using esophageal cancer Eca109 and KYSE150 cells as the target cells. The rate of inhibition and half-inhibitory concentration (IC₅₀) were measured by the CCK-8 method; clone formation ability was measured by clone formation assay; the changes in reactive oxygen species (ROS) were detected by flow cytometry; the ultrastructure of the cells was observed by transmission electron microscopy; the distribution of intracellular iron particles was observed by Prussian blue staining; the expression of glutathione peroxidase 4 (GPX4) was detected by immunofluorescence staining; the scratch assay was used to detect the cell migration ability; the Transwell assay was used to detect the cell invasion ability; and western blotting was used to detect E-cadherin, Slug and N-cadherin protein expression in the cells. The results show that realgar inhibited the proliferation of Eca109 and KYSE150 cells in a time- and concentration-dependent manner, and the IC₅₀ of Eca109 and KYSE150 cells was 64.297 and 51.337 μmol‧L^-1, respectively. Compared with the control group, many mitochondria in the cytoplasm of Eca109 and KYSE150 cells in the realgar 2IC₅₀ group were swollen and blue-stained particles of different sizes and amounts were found, and ROS fluorescence intensity was significantly increased while GPX4 protein expression was significantly reduced (P < 0.01). Compared with the realgar group, the proliferation, migration, membrane penetration and Slug and N-cadherin protein expression were significantly increased, and the cell inhibition rate and E-cadherin protein expression were significantly decreased in Eca109 and KYSE150 cells in the realgar+ZVAD-FMK group (P < 0.05). The proliferation, migration, membrane penetration and Slug and N-cadherin protein expression were significantly decreased, and the cell inhibition rate and E-cadherin protein expression were significantly increased of Eca109 and KYSE150 cells in the realgar +erastin group (P < 0.05). The above results show that realgar inhibited the proliferation of Eca109 and KYSE150 cells and induced partial ferroptosis in the cells, and the proliferation and metastasis effects of realgar on esophageal cancer cells may work through partial ferroptosis pathway activation.

The purpose of this study was to explore the mechanism of Sanzi Yangqin decoction (SYD) from the perspective of hypothalamic metabonomics to prevent the development of prehypertension (PHT) and excessive phlegm-dampness syndrome in rats. WKY rats were randomly divided into a normal group, a model group and a SYD group. The high-fat + high-salt method was used to induce PHT and excessive phlegm-dampness syndrome in the model and SYD rats. Different doses of SYD were used as an intervention over 8 weeks. Food consumption, weight, height, blood lipid and blood pressure were recorded for each group. The results show that, compared with the model group, the diet, weight and blood pressure of rats in SYD group decreased. Metabolites in the hypothalamus which differed in their expression between the three groups were identified and analyzed by LC-MS and the metabolic pathways were then determined. Fourteen metabolites in the hypothalamus were down-regulated and were mainly related to amino acid metabolism, glutathione metabolism, sphingolipid metabolism and glyceride metabolism, suggesting that SYD might alter hypothalamic dietary behavior, thereby affecting amino acid metabolism and energy metabolism. This study was approved by the Animal Ethics Review Committee of Shandong University of Traditional Chinese Medicine (approval number: SDUTCM20211103001).

By means of several chromatographic techniques, such as silica gel, Sephadex LH-20 and semi-preparative high performance liquid chromatography (HPLC), two caryolane-type sesquiterpenoids were isolated from the petroleum ether soluble fraction of the aerial parts of Pogostemon cablin (Blanco) Benth., and their structures were identified as caryolanol A (1) and 1α-ethoxy-9α-hydroxy-senecrassane (2) by various spectroscopic methods, including NMR and HR-ESI-MS. Compounds 1 and 2 were a pair of C₅ epimers, and compound 1 was a new compound. In addition, the anti-influenza virus activity of compounds 1 and 2 was evaluated.

Identification of metabolites of Danshen-Honghua herb pairs in isolated rat intestinal flora based on HPLC-Q-TOF-MS/MS technique. By incubating enterobacteria in isolated rats as well as inactivated enterobacteria in the incubation solution. The extracts of Danshen-Honghua herb pairs were added separately and co-incubated under anaerobic conditions. Animal experiments and protocols were approved by the Laboratory Animal Ethics Committee of Shaanxi University of Traditional Chinese Medicine (approval number: TCM-2020-030-E05). A total of 14 compounds, including 5 prototypes and 9 metabolites, were identified in the isolated rat intestinal incubation fluid. In contrast, no metabolites were detected in the inactivated enterobacterial fluid, except for the prototype component. The results showed that the main components of the Danshensu, salvianolic acid B, rosmarinic acid, lithospermic acid, and hydroxysafflor yellow A, could be metabolized by the intestinal flora, and these active ingredients were mainly metabolized in the rat intestinal flora in isolation by hydroxylation, decarboxylation, deoxygenation, decarboxylation and dehydration in phase Ⅰ, sulfate esterification and methylation in phase Ⅱ. This proved that the Danshen-Honghua herb pair could be transformed into various metabolites by the action of rat intestinal flora, further clarifying the role of intestinal flora in the metabolic transformation of the active ingredients of Chinese medicine and laying the foundation for perfecting the potent substances of the pair.

We predicted the anti-ulcerative colitis (UC) mechanism of Fructus Amomi based on network pharmacology. The anti-UC activity of Fructus Amomi were investigated by in vivo animal experiment, and the active components of Fructus Amomi were obtained through TCMSP, PubChem database and literature research. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of Institute of Materia Medica of Chinese Academy of Medical Sciences. The potential targets of the active components and UC were predicted by SwissTargetPrediction, GeneCards and TTD databases. The protein-protein interaction (PPI) network was constructed by String database and Cytoscape software was used to construct a visual network of active component-disease target and perform topological analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using Metascape platform. The molecular docking of key components and core targets was carried out by Sybyl X software. We screened out a total of 12 active components and 189 disease-component overlapping targets. Enrichment analyses obtained 227 related GO items and 168 signaling pathways. According to the results of molecular docking, most active components of Fructus Amomi showed good affinity with the JAKs receptor family. Furthermore, Western blot results verified that Fructus Amomi could effectively inhibit JAK/STAT signaling pathway, indicating that Fructus Amomi might exert the anti-UC activity by regulating JAK/STAT signaling pathway.

Acute lung injure (ALI) is a severe diffused lung disease, which is caused by pathogen-induced infections, inhalation of irritates, and so on. It could lead to acute respiratory distress syndrome (ARDS). Long oil (LO) is a lipidic mixture extracted from multiple medicinal animals and plants. It has been used for clinical wound repair. Here, an O/W LO emulsions (LOE) was prepared, which was composed of LO, Tween-80, propylene glycol, xanthan gum, and water. The droplet size of LOE was 671.63 ± 7.21 nm, and the zeta potential was-17.8± 1.26 mV. The size of LOE was small and homogenous, and the stability was satisfied. The aerosols had an aerodynamic diameter of 2.25 ± 0.05 μm after atomization of LOE with a vibrating screen atomizer, where the percentage of particle sizes within 1-5 μm was 81.40%, indicating effective deep lung deposition and suitable pulmonary inhalation. The safe dose of LOE was high to 12.50 μg·mL^-1 on human embryonic lung fibroblast MRC-5 cells. In the range of 0.02-2.50 μg·mL^-1 of LOE, the proliferation and migration of mouse fibroblast L929 cells were improved. Animal experiments were approved by the Ethics Committee of Institute of Radiation Medicine, Academy of Military Medical Sciences, and the experiments were conducted by relevant guidelines and regulations. No significant toxicity was observed after intratracheal (i.t.) administration of LO (3.25 mg·kg^-1) to mice. Mouse i.t. administration of LOE remarkably attenuated lung injury induced by lipopolysaccharide with mitigations of inflammatory factors (tumor necrosis factor-α, interleukin-6) and total proteins. LOE is a promising inhaled formulation for the treatment of ALI.

We have identified anti-inflammatory quality markers (Q-markers) of Jiangzhenxiang. The chemical components of Jiangzhenxiang were identified by mass spectrometry and the substances that contribute to its anti-inflammatory activity, their targets and signaling pathways were analyzed by network pharmacology to identify potential Q-markers of the anti-inflammatory action of Jiangzhenxiang. The potential Q-markers were verified by high performance liquid chromatography, and in vitro experiments verified the anti-inflammatory activity and the target of the potential Q-markers. The experimental scheme was approved the Guangxi University of Chinese Medicine Institutional Animal Ethical and Welfare Committee. The results show that 31 chemical components were identified by mass spectrometry from the Jiangzhenxiang extract. Through network pharmacological screening, 727 component targets, 422 disease targets and 110 targets including prostaglandin G/H synthase 2 (PTGS2) were obtained. These targets were mainly enriched in 498 biological processes including inflammatory response and response to lipopolysaccharide, with 101 pathways that included the TNF signaling pathway, toll-like receptor signaling pathway and others. Isorhamnetin, formononetin, naringenin, glycitein, ursolic acid and oleanolic acid were detected by high performance liquid chromatography. Jiangzhenxiang medicated serum and 6 components thereof could significantly reduce the content of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-α in RAW264.7 cells induced by lipopolysaccharide (P < 0.01 or P < 0.05). These six components are regarded as the potential anti-inflammatory Q-markers of Jiangzhenxiang. Isorhamnetin was screened and verified from the 6 potential Q-markers as an inhibitor of PTGS2 by molecular docking and in vitro cyclooxygenase 2 (COX-2) activity assay. The half-inhibitory concentration of isorhamnetin was 9.55μmol·L^-1. In summary, extracts of Jiangzhenxiang showed significant in vitro anti-inflammatory actions. The anti-inflammatory mechanism of Jiangzhenxiang appears to be related to regulation of TNF signaling pathway and Toll-like receptor signaling pathway meditated by IL-6, RAC alpha serine/threonine protein kinase, PTGS2 and other targets. Isorhamnetin, formononetin, naringenin, glycitein, ursolic acid and oleanolic acid could be regarded as the Q-markers of Jiangzhenxiang. Isorhamnetin appears to act as a COX-2 inhibitor.

A fungal strain WXF1904, was isolated from a starfish sample corrected in the South China Sea. According to its internal transcribed spacer (ITS) analysis, the strain was identified as a member of the genus Aspergillus and designated as Aspergillus sp. WXF1904. One new isocoumarin containing halogennamed 6-chloro-5, 7-dihydroxy-3, 8-dimethylisocoumarin (1), along with six known compounds pilobolusate (2), p-hydroxybenzaldehyde (3), methyl orsellinate (4), catechol (5), vanillic acid (6), and wasabidienone E (7), were isolated from the cultures of Aspergillus sp. WXF1904 by silica gel column chromatography, ODS gel column chromatography, and high performance liquid chromatography (HPLC). Their structures were elucidated by high resolution mass spectrometry (HR-MS), nuclear magnetic resonance (NMR) as well as literature comparison. The new compound 5-chloro-6, 7-dihydroxy-3, 8-dimethylisocoumarin and compound 2 showed weak acetylcholinesterase inhibitory activity.

Galli Gigerii Endothelium Corneum (GGEC) is a commonly used traditional Chinese medicine for digestion. Its odor is unpleasant, which decreases children's compliance with taking this traditional medicine. Traditional processing methods utilize heat processing methods such as stir-frying and vinegar processing to deodorize the medicine, but this affects the activity of digestive enzymes, so there is a need to find a new method for removing the fishy odor while retaining the beneficial effect of GGEC. Here we have developed the use of supercritical CO₂ low-temperature fluid extraction to eliminate the odor while retaining the medicinal benefits. Headspace-solid-phase microextraction-gas chromatography-triple quadrupole mass spectrometry (HS-SPME/GC-QQQ-MS/MS) combined with the gas activity value method was used to determine compositional differences in the product before and after supercritical CO₂ extraction and separation. Then, based on the sensory evaluation of volunteers, combined with the analysis of volatile components, the fishy odor intensity and the types of fishy odorants were compared between the raw product, stir-fried product, vinegar product and the supercritical CO₂ extract. Pepsin and amylase activity were used to compare the differences in the digestive enzyme activities with the four forms of GGEC, and Fourier transform infrared spectroscopy (FT-IR) was used to compare the differences in the structure. We compared the content of total amino acids, digestive amino acids and bitter amino acids, and an animal model of delayed gastric emptying in mice with soybean oil, based on a phenol red indicator, was used to determine differences in the efficacy of gastric emptying in vivo. The results show that the fishy odor of GGEC powder is significantly reduced after supercritical extraction, and the substances that contribute the fishy odor are only 12.8% of the raw material. The results of FT-IR analysis show that the supercritical extract of GGEC is not changed in its material structure compared with the raw product. The digestive enzyme activity titers showed that amylase and pepsin activity in the raw products are about 3.9 and 1.4 times higher than those of stir-fried products and vinegar products. The activity titers of amylase and pepsin in the supercritical CO₂extracts are about 2.7 and 1.3 times higher than those of stir-fried products and vinegar products, and there was no significant difference in the content of digestive-promoting amino acids in the four types of GGEC. The in vivo validation experiment showed that the average gastric emptying rates of the mice in the raw product group, the supercritical extract group, the stir-fried product group, and the vinegar product group were 69%, 59%, 40% and 51%, respectively. Compared with the stir-frying method and the vinegar-simmering method, the supercritical CO₂ fluid extraction method retained the gastric emptying effect of GGEC. In general, the supercritical CO₂ fluid low-temperature extraction method removes the fishy odor of GGEC as compared with the traditional stir-frying method and vinegar method, and retains the biologically active components and the effect on digestion of GGEC.

We have established a quantitative analysis of multi-components by single marker method (QAMS) for the simultaneous determination of apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, apigenin, hydroxygenkwanin, and genkwanin in Qutan Zhike Granules. The chromatographic column used was an Agilent EC-C18(150 mm × 4.6 mm, 4 μm), the mobile phase was methanol-0.15% phosphoric acid solution (gradient elution), and the detection wavelength was 338 nm. Apigenin was chosen as the internal reference standard, the relative correction factors for the six components were determined by multi-point correction method and included apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, hydroxygenkwanin and genkwanin. According to the two-point correction method combined with the relative retention time correction of the components to be tested, the peak location was determined. The contents of these seven compounds in 10 batches of Qutan Zhike Granules samples were determined with relative correction factors, and the relative error (RE) was used to compare the results to that of the external standard using the External Standard Method to verify the accuracy of QAMS. The relative correction factors for apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, hydroxygenkwanin and genkwanin were 1.762 8, 2.310 4, 1.898 4, 1.282 8, 1.191 3 and 1.066 9, respectively. RSDs of the relative correction factors were all lower than 3%. The peaks for each constituent were accurately located by the two-point correction method combined with relative retention time correction, and the predicted retention time was close to the actual retention time. The relative error of the contents by QAMS and determined by ESM in 10 batches of Qutan Zhike Granules were between -5% and 5%. This content determination method can be used for the simultaneous determination of seven components in Qutan Zhike Granules.