Background: Cerebral ischemic stroke (CIS) constitutes a serious and common neuropsychiatric complication. Modified Buyang Huanwu (MBYHW) Decoction, a formula incorporating herbs that promote blood circulation, resolve stasis, tonify Qi, and dredge collaterals, has been shown to improve cerebral circulation and function. However, its comprehensive chemical constituents and underlying pharmacological mechanisms remain incompletely elucidated. Methods: The chemical components of MBYHW were comprehensively examined using UPLC-Q-TOF/MS E . Metabolic analysis was performed on rat plasma after oral administration to identify prototype components and potential metabolites. Network pharmacology approaches, including protein-protein interaction network analysis, Gene Ontology enrichment, and Kyoto Encyclopedia of Genes and Genomes pathway analyses, and molecular docking, were employed to evaluate the antineuroinflammatory components of MBYHW Decoction for CIS treatment. The in vivo efficacy was further validated using a middle cerebral artery occlusion (MCAO) rat model. Results: A total of 190 compounds were identified in MBYHW Decoction in vitro , while 33 prototype components were detected and identified in plasma. Metabolic analysis combined with in vitro antineuroinflammatory evaluation confirmed 5 active components, thereby clarifying the pharmacological substance basis of MBYHW Decoction for CIS treatment. Pharmacodynamic assessments demonstrated that MBYHW Decoction significantly promoted cell survival and inhibited the release of pro-inflammatory factors in BV-2 cells induced by oxygen-glucose deprivation. Specifically, the 5 active compounds—stachydrine hydrochloride, leonurine hydrochloride, albiflorin, salvianolic acid B, and paeoniflorin—significantly suppressed the release of tumor necrosis factor-alpha and interleukin-6 in oxygen-glucose deprivation-induced BV-2 cells. Furthermore, in MCAO rats, MBYHW Decoction ameliorated neurological deficits and reduced cerebral infarct volume, further confirming its therapeutic efficacy in vivo. Conclusion: The chemical components of MBYHW Decoction and their metabolites in rat plasma were comprehensively characterized using UPLC-Q-TOF/MS E . A multidimensional component-disease-target-pathway network was subsequently constructed. Integrated analysis incorporating protein-protein interaction network, Gene Ontology enrichment, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and molecular docking revealed that components, including stachydrine hydrochloride, leonurine hydrochloride, albiflorin, salvianolic acid B, and paeoniflorin are pivotal to the therapeutic effects against CIS. These findings were substantiated by in vitro anti-inflammatory assays and in vivo efficacy validation using the MCAO model.

Introduction:Wuji Wan (WJW) is a classical Chinese formula traditionally prescribed for diarrhea/dysentery and abdominal pain. In ulcerative colitis (UC), inflammatory diarrhea reflects not only mucosal inflammation but also inflammation-linked disruption of epithelial electrolyte and water handling, highlighting membrane transport as a mechanistic bridge between symptom burden and immune activation. However, it remains unclear whether WJW confers therapeutic benefit in UC and whether any benefit is accompanied by coordinated regulation of membrane-transport-linked pathways. This study therefore asked whether WJW shows therapeutic effects in a UC model and whether these effects are accompanied by changes in epithelial Na+/Cl- transport and water-channel programs and by modulation of the T-cell-linked potassium channel Kv1.3. Methods:We investigated this question in a mouse model of DSS-induced colitis (3% dextran sulfate sodium). Our assessment included disease activity index (DAI) scores, histopathological analysis, ELISA, Western blotting, untargeted metabolomics, and whole-cell patch-clamp electrophysiology. Results:WJW significantly ameliorated DSS-induced colitis, as reflected by improved colonic pathology and partial normalization of DSS-associated serum metabolic perturbations. Untargeted metabolomics highlighted transport-related pathways. WJW increased/normalized the expression of key epithelial transport proteins involved in Na+/Cl- absorption and water handling, including sodium/hydrogen exchanger 3 (NHE3), epithelial sodium channel (ENaC), downregulated in adenoma (DRA), aquaporin-3 (AQP3), and aquaporin-8 (AQP8). In parallel, WJW reduced IL-6, IL-17A, and IFN-γ and dampened ERK/NF-κB pathway activation. WJW also reduced colonic Kv1.3 protein expression, and WJW-containing plasma directly inhibited Kv1.3 currents in Jurkat T cells. Conclusion:WJW ameliorated DSS-induced colitis and was accompanied by coordinated modulation of epithelial and immune membrane-transport-linked readouts.

The genus Fusarium is a prolific producer of chemically diverse natural products that exhibit a broad spectrum of biological activities, including antibacterial, insecticidal, and anticancer effects. In this study, bioassay-guided isolation from an endophytic fungus Fusarium iranicum YH6 led to the discovery of four previously undescribed cyclodepsipeptides, fusaristatins I-L (1-4), along with a known analogue fusaristatin A (5). Their planar structures were elucidated on the basis of 1D and 2D NMR, and HR ESI-MS comprehensive spectroscopic analyses. The relative and absolute configurations were rigorously assigned using advanced Marfey’s method, J-based configuration analysis, and DP4+ probability assessments. The biological activities of all isolated cyclodepsipeptides were evaluated, including anti-inflammatory activity in LPS-induced RAW264.7 macrophage cells, anti-proliferative activity against A549 and HCC827 cell lines, and antibacterial activity against Staphylococcus aureus. Compounds 1-5 exhibited potent anti-inflammatory activities by inhibiting NO production in LPS-stimulated RAW264.7 macrophages, with IC50 values of 5.7~20.5 µM. Fusaristatin K (3) displayed the most significant inhibition and down-regulated the expression of TLR4/NF-κB/NLRP3 signaling pathway to exert the antiinflammatory activity.

Two new polyketides, demethyl-epiremisporine B (1) and demethyl-coniochaetone A (2), together with four known polyketide compounds including diaportheone A (3), vertixanthone (4), peniphenone (5) and 3,8-dihydroxy-1-propylanthraquinone (6), were isolated from the endophytic fungus Phomopsis heveicola obtained from Sterculia lychnophora. The structures of the new metabolites were elucidated by NMR and HRESIMS analysis, and the absolute configuration of compound 1 was established by ECD spectroscopy. These compounds were evaluated for their antifungal activity against Candida albicans CMCC 98001 and Aspergillus niger R330. Compounds 4 and 6 showed significant antifungal activity on C. Albicans CMCC 98001, with the MIC of 3.125 μg/ml.

In this study, we employed a combination of genome mining and heteronuclear single quantum coherence (HSQC)-based small molecule accurate recognition technology (SMART) technology to search for fernane-type triterpenoids. Initially, potential endophytic fungi were identified through genome mining. Subsequently, fine fractions containing various fernane-type triterpenoids were selected using HSQC data collection and SMART prediction. These triterpenoids were then obtained through targeted isolation and identification. Finally, their antifungal activity was evaluated. As a result, three fernane-type triterpenoids, including two novel compounds, along with two new sesquiterpenes and four known compounds were isolated from one potential strain, Diaporthe discoidispora. Their structures were elucidated through analysis of high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopic data. The absolute configurations were determined using single-crystal X-ray diffraction analysis and electron capture detector (ECD) analysis. Compound 3 exhibited moderate antifungal activity against Candida albicans CMCC 98001 and Aspergillus niger.

Feature-based molecular networking analysis combined with bioassay-guided method was applied to afford two previously undescribed trichothecenes derivatives (1-2), together with eleven trichothecenes derivatives (3-13) and three β-resorcylic macrolides (14-16) from an endophytic fungus Fusarium equiseti Z8. The new structures were elucidated by using comprehensive spectroscopic analyses of NMR, HR-ESI-MS, and ECD calculations. MSn fragmentation of trichothecenes was also elucidated. All isolated compounds were evaluated the antiproliferative effects against A549, NCI-H1944, and NCI-H1650 human lung adenocarcinoma cell lines, as well as the antibacterial activity against Staphylococcus aureus. Most of the trichothecenes possessed significant antiproliferative activities, especially compounds 6 and 11 exhibited the most potent antiproliferative activities against the three tumor cells, with the IC50 values of 1.6-3.8 nM. Compounds 1-2 are relatively nontoxic to BEAS-2B human lung normal epithelial cells. Additionally, compound 14 displayed significant inhibition against S. aureus (MIC80: 4.17 μg/mL).

This study aims to characterize and identify the chemical constituents in 11 parts of Forsythia suspensa by using ultra-performance liquid chromatography-quadrupole time of flight-mass spectrometry(UPLC-Q-TOF-MS) combined with a self-established chemical constituent database, including leaves, flowers, fruits, green F. suspensa, old F. suspensa, and seeds. The quality attributes and differences of different parts of F. suspensa were evaluated by principal component analysis, partial least square discriminant analysis, and other stoichiometric methods. A total of 79 compounds were identified, including 13 phenylethanol glycosides, 10 lignans, 12 flavonoids, 10 organic acids, 14 terpenoids, and 20 other types of compounds. Among them, 34 compounds were the main variables of difference between the different parts of F. suspensa, and the content of each component was relatively higher in the leaves and green F. suspensa. The LPS-induced inflammation model of RAW264.7 cells was applied to study the anti-inflammatory activity of the extracts of the different parts of F. suspensa and the main constituents. The results show that the extracts of green F. suspensa, flower, twig, and stem exhibited anti-inflammatory activity, and the constituents such as forsythoside A, phyllyrin, phillygenin, and(+)-pinoresinol-β-D-glucopyranoside could significantly inhibit anti-inflammatory activity released by NO. The chemical constituent in different parts of F. suspensa is analyzed comprehensively, and the anti-inflammatory activity is evaluated in this study, which provides a reference for the development and comprehensive utilization of F. suspensa resources.

This study focused on the bioactive secondary metabolites of an endophytic fungus Aspergillus sp. CCH-1E from Catharanthus roseus. The secondary metabolites from Aspergillus sp. CCH-1E were isolated by using various chromatographic methods [such as normal-phase and reversed-phase chromatography and high-performance liquid chromatography(HPLC)], and their structures were identified by various spectroscopic methods [e.g., ultraviolet(UV) spectroscopy, infrared(IR) spectroscopy, nuclear magnetic resonance(NMR) spectroscopy, and high-resolution electrospray ionization mass spectrometry(HR-ESI-MS)]. Twelve compounds were yielded and identified from Aspergillus sp. CCH-1E, which are chermesinone H(1), chermesinone I(2), chermesinone B(3), 8,11-didehydrochermesinone B(4), chermesinone C(5), chermesinone A(6), chevalone B(7), barbacenic acid(8), 3,6,8-trihydroxy-3,5,7-trimethyl-3,4-dihydroisocoumarin(9), 5-hydroxy-2-methoxy-7-methyl-1,4-naphthoquinone(10), 1-hydroxy-6,8-dimethoxy-3-methylanthracene-9,10-dione(11), and 7-drimen-9α,11,12-triol(12). Among them, compounds 1 and 2 are new compounds. The growth inhibition effects of all compounds were evaluated against non-small cell lung cancer cell lines A549 and NCI-H1650, as well as human cervical cancer cell line HeLa by using methylthiazolyldiphenyl-tetrazolium bromide(MTT). Compound 7 significantly inhibited the growth of three tumor cells with the IC_(50) values of 1.22-2.43 μmol·L~(-1), respectively. Compounds 1-6 showed moderate cell growth inhibition with the IC_(50) values of 16.24-35.28 μmol·L~(-1).

This study investigated the chemical and biological activity of the secondary metabolites from an endophytic fungus Fusa-rium solani MBM-5 of Datura arborea. A total of six alkenoic acid compounds, including a new compound and five known ones, were isolated from the ethyl acetate extract of F. solani MBM-5 by using the chromatographic methods(open ODS column chromatography, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC). The structures of the compounds were identified by using their physical and chemical data, spectroscopic methods(UV, IR, NMR, and HR-ESI-MS), and Mosher's reaction, which were fusaridioic acid E(1), fusaridioic acid C(2), fusaridioic acid A(3), L660282(4), hymeglusin(5), and hymeglnone(6). Compound 1 is new. MTT assay and Griss method were used to evaluate the growth inhibition of all the compounds against two tumor cells, as well as their influence and anti-inflammatory action on the release of NO from LPS-induced RAW264.7 cells. The results showed that compound 5 had strong growth inhibition activity against A549 and HepG2 cell lines, with IC_(50) values of 4.70 and 13.57 μmol·L~(-1), respectively. Compounds 1 and 6 significantly inhibited the release of NO from LPS-induced RAW264.7 cells, with IC_(50) values of 77.00 and 70.33 μmol·L~(-1), respectively.

Lignin, a major wood component, is the key limiting factor for wood conversion efficiency. Its biosynthesis is controlled by transcriptional regulatory networks involving transcription factor (TF)-DNA interactions. However, the epigenetic mechanisms underlying these interactions in lignin biosynthesis remain largely unknown. Here, using yeast one-hybrid, chromatin immunoprecipitation, and electrophoretic mobility shift assays, we identified that PtrbZIP44-A1, a key wood-forming TF, directly interacts with the promoters of PtrCCoAOMT2 and PtrCCR2, genes involved in the monolignol biosynthetic pathway. We used yeast two-hybrid, bimolecular fluorescence complementation, biochemical analyses, transient and CRISPR-mediated transgenesis in Populus trichocarpa to demonstrate that PtrHDA15, a histone deacetylase, acts as an epigenetic inhibitor and is recruited by PtrbZIP44-A1 for chromatin histone modifications to repress PtrCCoAOMT2 and PtrCCR2, leading to reduced lignin deposition. In transgenic lines overexpressing PtrbZIP44-A1 or PtrHDA15, histone acetylation at the promoters of PtrCCoAOMT2 and PtrCCR2 decreased, reducing their expression and lignin content. Conversely, in loss-of-function ptrbzip44-a1 and ptrhda15 mutants, histone acetylation levels at PtrCCoAOMT2 and PtrCCR2 promoters increased, enhancing target gene expression and lignin content. Our study uncovered an epigenetic mechanism that suppresses lignin biosynthesis. This finding may help fill a knowledge gap between epigenetic regulation and lignin biosynthesis during wood formation in Populus.

Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities.Methods: The AlCl 3 colorimetric method and Prussian Blue assay were used to determine the contents of total flavonoids and total phenolic acids in extraction layers, and the bioactive layers was screened through anti-oxidative activity in vitro. The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC BEH C 18 column（2.0 mm × 150 mm, 5 μm） were used to identify the ingredients. And anti-oxidative ingredients were screened by off-line UPLC-QTOF-MS/MS-free radical scavenging. The ameliorative role of it was further evaluated in a high-fat, streptozotocin-induced type 2 diabetic rat model and the study was carried out on NADPH oxidase（PDB ID: 2CDU） by molecular docking.Results: Combined with the results of activity screening in vitro, the anti-oxidative part was identified as the ethyl acetate layer. A total of 24 chemical constituents were identified by liquid chromatographymass spectrometry in the ethyl acetate layer and 13 main anti-oxidative active constituents were preliminarily screened out through off-line UPLC-QTOF-MS/MS-free radical scavenging. In vivo experiments showed that flowers of X. sorbifolia could significantly reduce the blood glucose level of diabetic mice and alleviate liver cell damage. Based on the results of docking analysis related to the identified phytocompounds and oxidase which involved in type 2 diabetes, quercetin 3-O-rutinoside, kaempferol-3-O-rhamnoside, isorhamnetin-3-O-glucoside, and isoquercitrin showed a better inhibitory profile.Conclusion: The ethyl acetate layer was rich in flavonoids and phenolic acids and had significant anti-oxidant activity, which could prevent hyperglycemia. This observed activity profile suggested X. sorbifolia flowers as a promising new source of tea to develop alternative natural anti-diabetic products with a high safety margin.

Tyrosine-decahydrofluorene derivatives are a class of hybrid compounds that integrate the properties of polyketides and nonribosomal peptides. These compounds feature a [6.5.6] tricarbocyclic core and a para-cyclophane ether moiety in their structures and exhibit anti-tumor and anti-microbial activities. In this study, we constructed the biosynthetic pathway of xenoacremones from Xenoacremonium sinensis ML-31 in the Aspergillus nidulans host, resulting in the identification of four novel tyrosine-decahydrofluorene analogs, xenoacremones I-L (1-4), along with two known analogs, xenoacremones A and B. Remarkably, compounds 3 and 4 contained a 12-membered para-cyclophane ring system, which is unprecedented among tyrosine-decahydrofluorene analogs in X. sinensis. The successful reconstruction of the biosynthetic pathway and the discovery of novel analogs demonstrate the utility of heterologous expression strategy for the generation of structurally diverse natural products with potential biological activities.

Two new dipyrroloquinones, namely talaroterreusinones A (1) and B (2), together with four known secondary metabolites, terreusinone A (3), penicillixanthone A (4), isorhodoptilometrin (5), and chrysomutanin (6), were isolated from the solid culture of the endophytic fungus Talaromyces sp. by integrating mass spectrometry-based metabolic profiling and a bioassay-guided method. Their planar structures and stereochemistry were elucidated by comprehensive spectroscopic analysis including NMR and MS. The absolute configuration at C-1″ of terreusinone A (1) was established by applying the modified Mosher’s method. Compounds 1–6 were evaluated for anti-inflammatory activity and cytotoxicity. As a result, 1–3 inhibited the LPS-stimulated NO production in macrophage RAW264.7 cells, with IC50 values of 20.3, 30.7, and 20.6 µM, respectively. Penicillixanthone A (4) exhibited potent cytotoxic activity against Hep G2 and A549 cell lines, with IC50 values of 117 nM and 212 nM, respectively, and displayed significant antitumour effects in A549 cells by inhibiting the PI3K-Akt-mTOR signalling pathway.

Aristolochic acids (AAs) are a group of nitrophenanthrene carboxylic acids present in many medicinal herbs of the Aristolochia genus that may cause irreversible hepatotoxicity, nephrotoxicity, genotoxicity and carcinogenicity. However, the specific profile of AAs and their toxicity in Aristolochia plants, except for AAs Ι and ΙΙ, still remain unclear. In this study, a total of 52 batches of three medicinal herbs belonging to the Aristolochia family were analyzed for their AA composition profiles and AA contents using the UPLC-QTOF-MS/MS approach. The studied herbs were A. mollissima Hance (AMH), A. debilis Sieb.etZucc (ADS), and A. cinnabaria C.Y.Cheng (ACY). Chemometrics methods, including PCA and OPLS-DA, were used for the evaluation of the Aristolochia medicinal herbs. Additionally, cytotoxicity and genotoxicity of the selected AAs and the extracts of AMH and ADS were evaluated in a HepG2 cell line using the MTT method and a Comet assay, respectively. A total of 44 AAs, including 23 aristolochic acids and 21 aristolactams (ALs), were detected in A. mollissima. Moreover, 41 AAs (23 AAs and 18 ALs) were identified from A. debilis Sieb, and 45 AAs (29 AAs and 16 ALs) were identified in A. cinnabaria. Chemometrics results showed that 16, 19, and 22 AAs identified in AMH, ADS, and ACY, respectively, had statistical significance for distinguishing the three medicinal herbs of different origins. In the cytotoxicity assay, compounds AL-BΙΙ, AAΙ and the extract of AMH exhibited significant cytotoxicities against the HepG2 cell line with the IC50 values of 0.2, 9.7 and 50.2 μM, respectively. The results of the Comet assay showed that AAΙ caused relatively higher damage to cellular DNA (TDNA 40–95%) at 50 μM, while AAΙΙ, AMH and ADS extracts (ranged from 10 to 131 μM) caused relatively lower damage to cellular DNA (TDNA 5–20%).

The secondary metabolites of Fusarium sporotrichioides, an endophytic fungus with anti-tumor activity isolated from Rauvolfia yunnanensis Tsiang, were investigated. Five trichothecenes, including one previously undescribed metabolite, were isolated and identified. Their structures were elucidated by means of extensive spectroscopic methods; the absolute configuration of compound 1 was determined by the ECD method. Surprisingly, 8-n-butyrylneosolaniol (3) exhibited stronger anti-tumor activity than T-2 toxin against Huh-7 cell line, with an IC50 value of 265.9 nM. 8-n-butyrylneosolaniol (3) promoted apoptosis induction in Huh-7 cells. Moreover, cell cycle analysis showed that cell cycle arrest caused by 8-n-butyrylneosolaniol (3) at the G2/M phase resulted in cell proliferation inhibition and pro-apoptotic activity. Further studies showed a significant decrease in mitochondrial membrane permeabilization and a significant increase in ROS generation, which led to the activation of caspase cascades and subsequent cleavage of PARP fragments. In conclusion, 8-n-butyrylneosolaniol (3) induced cell apoptosis in Huh-7 cells via the mitochondria-mediated apoptotic signaling pathway, which could be a leading compound for anti-tumor agents.

Three new ergosterols, colletosterols A-C (1-3), together with two known analogues 4 and 5, were isolated from the endophytic fungus Colletotrichum magnisporum associated with the leaves of Rauvolfia verticillata by a bioassay-guided fractionation method. The new structures were elucidated on the basis of extensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. All the ergosterols were evaluated for their cytotoxic activities against A549 and HeLa cell lines. Compounds 1-3 exhibited notable cytotoxicity with the IC50 values of 3.76-11.18 mu M.

As a classic prescription, Wuji Pills is composed of Coptidis Rhizoma, Euodiae Fructus Preparata, and stir-fried Paeo-niae Radix Alba at the ratio of 6∶1∶6. The practical application of it is limited compared with other famous Chinese medicine prescriptions. Only one company produces Wuji Pills in China. In this study, ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to analyze and identify 26 identical compounds from Wuji Pills and drug-containing plasma of rats. Based on these components, 46 potential targets were screened out with network pharmacology methods, followed by the component-target network construction, Gene Ontology(GO) term enrichment, Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment, and disease prediction. It was concluded that Wuji Pills acted on core targets such as PTGS2, PTSG1, NCOA2, HSP9 OAD1, and RXRA through magnoflorine, hydroxyevodiamine, daucosterol, and berberine and exerted pharmacodynamic effects through various pathways such as calcium ion signaling pathway, phosphatidylinositol-3-kinase-protein kinase B(PI3 K-Akt) signaling pathway, and vascular endothelial growth factor(VEGF) signaling pathway. Thus, Wuji Pills has therapeutic potential for Alzheimer's disease, diabetes mellitus, myocardial ischemia, and other diseases in addition to the conventional disease(irritable bowel syndrome, IBS). The above research results can provide a reference for the comprehensive interpretation of the pharmacodynamic basis of Wuji Pills and the expansion of clinical application. At the same time, a lot of components in serum and the in vivo transformed and metabolized components of Wuji Pills have similar structure and relative molecular weight. In theory, these components may show additive effects and the competitive/antagonistic effects on the same target. According to the hypothesis of "additive effect of multiple components for a single target" in traditional Chinese medicine, multiple similar components may exert the additive effects on local targets. This study can partly prove the scientificity of this hypothesis and provide laboratory evidence.

Five novel tyrosine-decahydrofluorene analogues, xenoacremones D–H (1–5), each bearing a fused 6/5/6 tricarbocyclic core and a 13-membered para-cyclophane ring system, were isolated from the endophytic fungus Xenoacremonium sinensis. Compound 1 was a novel polyketide synthase–nonribosomal peptide synthetase (PKS–NRPS) tyrosine-decahydrofluorene hybrid containing a 6/5/6/6/5 ring system. Their structures were elucidated from comprehensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. All compounds were evaluated for their inhibitory activities on LPS-induced NO production in macrophages and their cytotoxicities against the NB4 and U937 cell lines. Compounds 3 and 5 exhibited potent anti-inflammatory activities in vitro. Compounds 1 and 3–5 displayed significant antiproliferative activity against the tumor cell lines (IC50 < 20 µM).