BACKGROUND:Climate change-induced heatstroke (HS) presents a critical global public health threat, necessitating the development of preventive strategies using natural, food-derived resources. Pogostemon cablin (patchouli) is a medicinal and edible dual-purpose resource that yields an essential oil (PEO), which serves as a core ingredient in traditional medicines and a promising functional food additive. However, the bioactive basis and mechanisms of PEO in maintaining physiological homeostasis against HS remain to be fully elucidated. This study assessed the protective effects of PEO on intestinal barrier function and immune homeostasis under HS conditions. RESULTS:Gas chromatography-mass spectrometry analysis identified 42 volatile constituents in PEO, predominantly sesquiterpenes and sesquiterpene alcohols. In an HS mouse model, oral administration of PEO accelerated thermoregulatory recovery and markedly attenuated hepatic and ileal structural damage. Specifically, PEO preserved intestinal barrier integrity by upregulating key tight junction proteins, including Zonula Occludens-1 and Occludin. Furthermore, PEO suppressed the systemic pro-inflammatory cytokine storm and modulated immune balance in heat-stressed RAW264.7 macrophages by inhibiting M1 polarization (iNOS, TNF-α) and promoting M2 markers (Arg1, IL-10). CONCLUSION:These results revealed that PEO serves as a potent effective component that protects against HS-induced injury by reinforcing intestinal barrier function and regulating immune homeostasis. These findings provide theoretical support for the development of PEO as a promising functional food or beverage additive for mitigating heat-related challenges. © 2026 Society of Chemical Industry.

Epimedium total flavonoid capsules (ETFCs), a traditional Chinese patent medicine derived from Epimedium brevicornu Maxim., have been used for centuries to treat primary osteoporosis and are associated with kidney yang deficiency symptoms. Based on the "kidney-brain axis" hypothesis, previous study demonstrated that ETFCs can improve the cognitive function of MCAO rats, and the mechanism may involve the inflammatory response. However, little is known about their therapeutic components and scientific connotations. This study systematically investigates the anti-neuroinflammatory material basis and the mechanisms of ETFCs, integrating UPLC-Q/ TOF-MS and UPLC-TQ-MS for chemical profiling, quantitative analysis of key components in rats ' plasma and brain, along with network pharmacology and activity verification. As a result, 65 components were identified. Metabolite profiling showed a total of 130 xenobiotics, highlighting the extensive metabolic transformations. Pharmacokinetic studies showed that primary flavonoid glycosides were quickly absorbed, while secondary glycosides exhibited slower absorption and elimination. In brain tissue, both prototype glycosides and secondary glycosides reached their peak concentrations rapidly, with Tmax between 0.25-1 h, while the prototype glycosides were eliminated after 8 h and secondary glycosides exhibited a secondary peak at 6-8 h. A network pharmacology study revealed that prototypes in blood may play a therapeutic role through targets including AKR1B1, PDE5A, and PTGS2 (COX-2). The LPS-induced BV-2 cell model confirmed ETFCs ' constituents showed inhibitory activities on pro-inflammatory cytokine secretion, mRNA expressions of IL-6, TNF-alpha, COX-2, iNOS and could down-regulate the protein expressions of COX-2 and iNOS. This research lays the foundation for the further development and utilization of ETFCs.

The anti-inflammatory phytochemical investigation of the leaves of Illicium dunnianum (I. dunnianum) resulted in the isolation of five pairs of new lignans (1-5), and 7 known analogs (6-12). The separation of enantiomer mixtures 1-5 to 1a/1b-5a/5b was achieved using a chiral column with acetonitrile-water mixtures as eluents. The planar structures of 1-2 were previously undescribed, and the chiral separation and absolute configurations of 3-5 were reported for the first time. Their structures were determined through comprehensive spectroscopic data analysis [nuclear magnetic resonance (NMR), high-resolution electrospray ionization mass (HR-ESI-MS), infrared (IR), and ultraviolet (UV)] and quantum chemistry calculations (ECD). The new isolates were evaluated by measuring their inhibitory effect on NO in lipopolysaccharide (LPS)-stimulated BV-2 cells. Compounds 1a, 3a, 3b, and 5a demonstrated partial inhibition of NO production in a concentration-dependent manner. Western blot and real-time polymerase chain reaction (PCR) assays revealed that 1a down-regulated the messenger ribonucleic acid (mRNA) levels of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), COX-2, and iNOS and the protein expressions of COX-2 and iNOS. This research provides guidance and evidence for the further development and utilization of I. dunnianum.

Piper longum L., a widely recognized Chinese traditional spice, is extensively used as dietary flavoring agent in Southeast Asia due to its unique taste. As a dual-purpose resource for both food and medicine, the fruits of P. longum have garnered significant attention for their diverse chemical constituents and notable physiological activities. In this study, the phytochemical investigation of the fruits of P. longum resulted in the isolation and identification of 20 structurally diverse piperidine amide alkaloids, including 4 new (1a/1b-4a/b) and 16 known analogues (5-20). The enantiomeric mixture of 1a/1b was successfully separated using a chiral column with acetonitrile-water as eluents, while chiral HPLC analysis confirmed that 4 existed as an enantiomeric mixture (4a/4b) without achieving separation. Their structures were elucidated through comprehensive spectroscopic data analysis (NMR, HRESIMS, IR, UV), electronic circular dichroism (ECD) and by comparison with literature data. Biological evaluations revealed that compounds 2, 3, 16, and 17 exhibited significant inhibitory effects on NO secretion in LPS-stimulated RAW264.7 cells, with IC50 values ranging from 9.9 to 15.76 μM, which were better than hydrocortisone (IC50 = 34.26 ± 2.20 μM). Furthermore, Western blot and real-time PCR assays demonstrated that 2 and 17 markedly downregulated the mRNA levels and the protein expression of iNOS. Notably, 17 partially inhibited the phosphorylation of p65 and reduced the protein expression of TLR4. Thus, the discovery of structurally diverse anti-inflammatory piperidine amide alkaloids from the fruits of P. longum in this study could benefit the further development and utilization of this plant.

Background Gout is a systemic metabolic disease with rising prevalence and complex etiology, yet its molecular mechanisms remain incompletely defined. Multi-omics Mendelian randomization (MR) enables causal prioritization of genes implicated in gout. Because the gut microbiota (GM) can modulate urate metabolism and inflammation, integrating microbiome analyses may help nominate potential therapeutic candidates. This study aimed to nominate candidate targets for gout by integrating multi-omics MR, clinical validation, GM analysis, and in-silico druggability assessment. Methods We performed multi-omics MR (pQTL/eQTL/mQTL) to prioritize genes with putative causal effects on gout risk. Candidate gene expression patterns were examined in a public single-cell RNA sequencing dataset to determine their distribution across immune cell subtypes and functional context. Associations between identified candidate genes and GM composition were evaluated using the MiBioGen database. Potential therapeutic interactions were predicted through the Comparative Toxicogenomics Database (CTD). Expression of candidate genes was validated in peripheral blood samples from 51 gout patients and 50 healthy controls by quantitative real-time polymerase chain reaction (qRT-PCR). Finally, molecular docking and molecular dynamics (MD) simulations were conducted to explore binding poses and stability between prioritized proteins and predicted drugs. Results MR analysis identified four gout-associated genes (BAIAP2, CD248, GCHFR, and ABHD14B). Among them, BAIAP2, CD248, and GCHFR showed significant causal relationships with specific gut microbial taxa in MiBioGen, whereas ABHD14B showed no GM associations. CTD-based drug prediction further highlighted three compounds—benzbromarone, calcitriol, and cyclosporine—potentially targeting these genes. In an independent cohort (51 gout vs. 50 controls), qRT-PCR confirmed GCHFR dysregulation consistent with MR estimates, and molecular docking plus MD simulations supported stable binding of benzbromarone to GCHFR, providing mechanistic plausibility. Conclusions This integrative framework convergently prioritizes GCHFR as a candidate for therapeutic investigation in gout and identifies CD248, BAIAP2, and ABHD14B as additional gout-related candidates. These results generate testable hypotheses for GCHFR-focused mechanistic and therapeutic studies, as well as further functional studies of the other genes.

Tianshu Capsule (TSC), a traditional Chinese medicine (TCM) formula derived from “Da Chuan Xiong Fang”, has a long history in migraine treatment. Studies have shown that TSC crude polysaccharides exhibit superior anti-migraine effects, but their chemical characteristics, in vivo pharmacokinetics, and distribution profiles remain uncharacterized. In this study, we elucidated the structures of a 1,4-glucan (TSCP-1a) and a novel pectin-like polysaccharide (TSCP-4) featuring a backbone of glucogalacturonan substituted by arabinans and arabinogalactan (AG) residues from TSC. In a nitroglycerin-induced migraine animal model, the homogeneous polysaccharide TSCP-4 significantly alleviated migraine symptoms, corresponding to the consistent bioactivity of TSC formula. To track its in vivo behavior, TSCP-4 was fluorescently labeled with cyanine 5.5 amine (C-TSCP4) and FITC (F-TSCP4). Near-infrared imaging showed that orally administered C-TSCP4 was absorbed in the small intestine and distributed to liver and kidney. Furthermore, the quantitative method for F-TSCP4 was applied to the pharmacokinetic analysis, revealing that F-TSCP4 exhibited favorable absorption characteristics and prolonged mean residence time (MRT). Conclusively, this study provides a detailed analysis of the fine structure and oral absorption properties of TSC-active homogeneous polysaccharides, offering valuable insights into the material basis of compound polysaccharides in TCM formulations.

A novel acidic polysaccharide, ATP-4, was isolated from Amomum tsao-ko and demonstrated immunomodulatory potential in the immunosuppressed mouse model and cellular assays. Structural characterization via monosaccharide analysis and gel permeation chromatography revealed ATP-4 as a heteropolysaccharide (4.23 × 104 Da), composed of Rha, GalA, Glc, Gal, Xyl, and Ara in a molar ratio of 9.03:52.37:7.89:12.34:6.47:11.89. Integrated analysis of conventional techniques (PMP-HPLC, GC-MS, and NMR) and UPLC-Orbitrap-MS/MS profiling of free radical-degraded components (AO-1/2/3) elucidated its unique repeating units, which are characteristic of a novel polysaccharide assembled through specific patterns between the HG regions and branched domains such as GRP, RGP, etc. In vitro and in vivo studies demonstrated ATP-4's dual mechanisms: direct immune cell activation and indirect gut microbiota modulation. Notably, active structural domains AO-2 (the nonlinear branched region of ATP-4) significantly induced cytokine production at a lower concentration, guiding activity-directed enzymatic modification of ATP-4. The optimized derivative, ATP-4e, maintained a nonlinear branching structure while demonstrating enhanced immunoregulatory capacity, improved conformational flexibility, and superior physicochemical properties compared to the native ATP-4. This study establishes an integrated degradation-analysis-modification paradigm for polysaccharide research and provides a methodological framework for elucidating structure-activity relationships of traditional Chinese medicine polysaccharides.

Background: Migraine is widely recognized as the third most prevalent medical condition globally. Tianshu capsule (TSC), derived from "Da Chuan Xiong Fang" of the Jin dynasty, is integral in the clinical treatment of migraine. However, the chemical properties and therapeutic mechanisms of TSC different portions remain unclear. Purpose: This study was designed to investigate the effects of TSC different portions (including small molecular TSCP-SM and polysaccharides TSC-P) on migraine and explore the underlying mechanisms. Study design and methods: First of all, migraine rats were established by nitroglycerin injection and treated with TSC, TSC-P, and TSC-SM. ELISA, qPCR, and immunofluorescence were used to evaluate the pharmacological effects on migraine rats. Secondly, UPLC-Q/TOF-MS and GC--MS were employed to detect the components of TSC-SM. PMP-HPLC, NMR, FT-IR, UV-Vis, AFM, and SEM were used for the chemical profiling of polysaccharides. Thirdly, the metabolic behavior profile of TSC-P was characterized by oral administrated fluorescence-labeled TSC-P and detected by NIRF imaging. Finally, the anti-migraine mechanisms were explored by determining the composition of gut microbiota, analyzing colonic short-chain fatty acids (SCFAs), and examining serum tryptophan-related metabolites. Results: Both small molecules (45 volatiles and 114 small molecules) and polysaccharides (including Glc, Ara, Gal, and Gal A) have exhibited effectiveness in alleviating migraine, and this efficacy is associated with reduced CGRP and iNOS levels, along with increased beta-EP expressions. Further mechanistic exploration revealed that small-molecules exhibited effectiveness in migraine treatment by exerting antioxidative actions, while polysaccharides demonstrated superior therapeutic effects in regulating 5-HT levels. By monitoring the metabolic behavior of polysaccharides with fluorescent labeling, it was observed that TSC-P exhibited poor absorption. Instead, TSC-P demonstrated its therapeutic effects by modulating the aberrations in gut microbiota (including Alloprevotella, Muribaculaceae_ge, and Ruminococcaceae_UCG-005), cecum short-chain fatty acids (such as isobutyric, isovaleric, and valeric acids), and serum tryptophan-related metabolites (including indole-3-acetamide, tryptophol, and indole-3-propionic acid). Conclusion: This research provides innovative insights into chemical composition, metabolic behavior, and proposed anti-migraine mechanisms of TSC from a polarity-based perspective, and pioneering an exploration focused on the polysaccharide components within TSC for the first time.

Four previously unreported diarylheptanoids (1a/1b-2a/2b), one undescribed sesquiterpenoid (8), one new diterpenoid (12), and twelve known analogs were isolated from the fruits of Alpinia oxyphylla. The structural elucidation of these compounds was achieved through a comprehensive analysis of spectroscopic data, single-crystal X-ray diffraction, electronic circular dichroism (ECD), and modified Mosher's method. Enantiomeric mixtures (1a/1b, 2a/2b, 3a/3b, 4a/4b, and 5a/5b) were separated on a chiral column using acetonitrile-water mixtures as eluents. Among them, compounds 3a/3b and 4a/4b were isolated as optically pure enantiomers in the initial chiral separation. Furthermore, most of the isolates were evaluated for their inhibitory effects against the production of nitric oxide (NO) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Interestingly, 2 and 4 showed significant inhibitory activities against NO production with IC50 values of 33.65 and 9.88 μmol·L-1 (hydrocortisone: IC50 34.26 μmol·L-1), respectively. Additionally, they also partially reduced the secretion of IL-6.

Four previously unreported phenylpropanoid glycosides (1–4), together with four known analogues (5–8), were isolated from the leaves of Illicium dunnianum. The structures of these new compounds were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR, IR, UV) and chemical methods. In addition, the neuroprotective activities of all the isolates were evaluated by measuring their cell viability in H2O2-induced OLN-93 cell injury model.

Background: Jin-Zhen oral liquid (JZOL), a well-known traditional Chinese medicine prescription (TCMp), has extensively been used to treat acute bronchitis in children more than four hundred years in China. However, the current quality control standard of JZOL is inadequate, posing challenges for its internationalization.Purpose: In this study, a Q-marker screening strategy based on multi-factor analysis was proposed to compre-hensively evaluate anti-inflammatory Q-markers of Jin-Zhen oral liquid (JZOL).Methods: Firstly, the chemical profile and the pharmacokinetics properties of multiple components in JZOL were characterized by UPLC-Q/TOF-MS and UPLC-QqQ-MS. By integrating the measurable and absorbed components, twenty-two components with structures accurately defined, were selected as candidate Q-markers of JZOL. Following that, a network connecting 22 components and targets closely associated with bronchitis was established. Afterwards, a multi-factor analysis mode was developed to balance the components' multiple characteristics, and screen out the anti-inflammatory Q-markers in JZOL. Finally, the anti-inflammatory activity evaluation was conducted by LPS-induced RAW 264.7 macrophages to prove the representativeness of anti-inflammatory Q-markers in JZOL.Results: As a result, a total of 92 components were characterized in JZOL and the pharmacokinetics properties of 11 bioactive components in vivo were further characterized. Then, an overlapping of 46 targets involved in the interactions of selected 22 candidate Q-markers and the regulation of bronchitis inflammation were collected. Subsequently, a multi-factor analysis was developed on 22 candidate anti-inflammatory Q-markers covering five factors, with the statistic KMO values of 0.645, and the P values of Bartlett's Test equal to 0.000. A total of seven ingredients (aloeemodin-8-O-beta-D-glucopyranoside, baicalin, chrysin-7-O-beta-D-glucuronide, oroxylin A 7-O-beta-D- glucuronide, wogonoside, chrysophanol-8-O-beta-D-glucopyranoside, and skullcapflavone II) were selected as Q -markers of JZOL, and the inhibitory effects of these candidate Q-markers on the secretion of inflammatory cytokes (NO, IL-6, IL-1 beta, and PGE2) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells were evaluated and confirmed.Conclusion: This study not only offers a fresh approach to uncovering Q-markers in the quality control research of TCMps but also identifies the suitable anti-inflammatory Q-markers for JZOL for the first time, with the potential to serve as a reference for existing quality control standards of JZOL.

One undescribed benzofuran derivative (illiciumphenolicacid A, 1) and one new phenolic glycoside (illiciumphenolicacid B, 2), together with six known compounds (3-8) were isolated from the leaves of Illicium dunnianum Tutcher. Their structures were elucidated by detailed spectroscopic data (UV, IR, HR-ESI-MS, 1D and 2D NMR). In addition, we determined the α-glucosidase inhibitory activity of the isolates in vitro using spectrophotometric methods. Compared with the positive control acarbose (IC50 306.2 ± 4.1 μM), compounds 1-8 were shown to be moderate potential α-glucosidase inhibitory activity with IC50 values in the range 380-655 μM.

The fruits of Alpinia oxyphylla have been used for centuries in China as both edible resources and traditional Chinese medicine. In order to identify structurally interesting and bioactive constituents from the fruits of A. oxyphylla, bioassay-guided fractionation and purification of the crude extracts were performed, which led to the isolation of 38 sesquiterpenoids, including six previously undescribed eremophilane sesquiterpenoids (1-6), six new cadinane sesquiterpenoids (23-24, 26-29), and 26 known analogues (7-22, 25 and 31-38). The structures of these compounds were elucidated by comprehensive spectroscopic data analysis, single crystal X-ray diffraction, quantum chemistry calculations (13C-NMR and ECD), and Mo2(OAc)4 reaction. Several of the isolated compounds (8, 13, 17, 18, 30, 31 and 35) showed moderate to strong inhibition of the secretion of cytokines (NO, TNF-α and IL-6) in LPS-stimulated BV-2 cells. Furthermore, western blot, immunofluorescence, and real-time PCR assays indicated that 18 could down-regulate the mRNA levels of TNF-α, IL-6, COX-2, and iNOS and the protein expression of COX-2 and iNOS. Meanwhile, 18 was able to partially inhibit the phosphorylation of ERK1/2, JNK, and p38. Thus, the discovery of structurally diverse anti-inflammatory sesquiterpenoids from the fruits of A. oxyphylla in this study could benefit the further development and utilization of this plant.

This study identified two homogeneous acidic polysaccharides from Gardeniae fructus, GJP50-3 and GJP50-4, which exhibited potential immunomodulatory activities in macrophage activation assays, via liquid-chip technology, and in a zebrafish model. Monosaccharide composition analysis and gel permeation chromatography revealed that GJP50-3 and GJP50-4 were composed of Rha, GalA, Glc, Gal, and Ara in specific ratios and had molecular weights of 91.5 kDa and 140.3 kDa, respectively. Based on FT-IR, GC-MS, and NMR analyses, these polysaccharides were identified as typical pectin polysaccharides with methylation degrees of 24.7 % and 21.4 %, respectively. The primary structures of GJP50-3 and GJP50-4 included linear HG domains and branched RG-I domains with arabinans and AG side chains. In vitro, GJP50-3 and GJP50-4 could stimulate NO release and increase the secretion of TNF-& alpha; in a RAW 264.7 macrophage model. Luminex liquid suspension chip detection revealed that GJP50-3 significantly promoted the secretion of multiple interleukins [IL-6, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13], TNF-& alpha;, and chemokines (G-CSF, GM-CSF, MCP-1 and RANTES). In vivo, these polysaccharides could also increase NO release and neutrophil count in a zebrafish model. These findings suggested that GJP50-3 and GJP50-4 might have the potential to be used as immunomodulators in the food and pharmaceutical industries.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Two undescribed labdane diterpenoids (5S,8S,9R,10S,11E)-8,17-epoxy-13,14-dinorlabd-11-en-13-one (1) and (5S,9R,10S,12E)-17-hydroxy-labd-7,12-dien-15(16)-olide (2), together with seven known sesquiterpenoids (3-9) and two known monoterpenoids (10-11) were isolated from the dried rhizome of Zingiber officinale. Their structures were elucidated by detailed spectroscopic data (IR, UV, HR-ESI-MS, 1D and 2D NMR), X-ray crystallographic and ECD analysis. Moreover, all the 11 compounds were tested for alpha-glucosidase inhibitory effects and 9 was found to exhibit stronger inhibitory effects at IC50 = 4.8 mu M against a positive control acarbose with IC50 = 414.6 mu M. [GRAPHICS] .

Two undescribed sesquiterpenoids, including one nor-eudesmane type (1) and one guaiane type (2), together with two known analogues (3-4) have been isolated and identified from the fruits of Alpinia oxyphylla. The structures of these new compounds were elucidated by extensive spectroscopic analyses (1D-, 2D-NMR, HRESIMS, IR, UV) and NMR calculations with DP4+ analysis. The anti-inflammatory activities of all isolates were evaluated by measuring their inhibitory effects on PGE2 production in LPS stimulated RAW 264.7 macrophages.

Abstract One new phenolic glycoside (1) and one new benzofuran derivative (2) were isolated from the leaves of Illicium dunnianum. The structures of these compounds were established by using comprehensive spectroscopic data analysis, including the 1D and 2D NMR, IR, HR-ESI-MS, electronic circular dichroism and comparison with literature data. All isolates were evaluated for the inhibition against the production of NO by LPS-stimulated RAW 264.7 macrophages. Graphical Abstract

Two new chemical constituents, japopenoid D (1), and japopenoid E (2), were isolated and identified from the flower buds of Lonicera japonica Thunb. The structures of these compounds were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR), and the absolute configurations of 1 and 2 were determined by comparison of their electronic circular dichroism (ECD) spectra with literature and theoretical calculation. The anti-inflammatory activities of the isolates were evaluated by measuring their inhibitory effects on PGE(2) and IL-6 production in LPS stimulated RAW 264.7 macrophages. As a result, compound 1 could reduce PGE(2) and IL-6 levels in LPS-activated RAW 264.7 macrophages with IC50 values of 6.78 and 42.07 mu M, respectively.