Huoxue San (HXS) is a traditional Chinese medicinal formulation widely used to treat bone fractures and blood stasis. Comprising seven herbs-Siphonostegia chinensis Benth, Kochia scoparia (L.) Schrad, Scutellaria barbata D.Don, Polygonum cuspidatum Sieb. et Zucc, Arisaema erubescens (Wall.) Schott, Phellodendron chinense Schneid, and Eupolyphaga sinensis Walker-HXS has been administered at Nanjing Chinese Medicine Hospital for over 50 years. It is effective in promoting fracture healing, supporting soft tissue repair, and rarely causing adverse reactions such as skin allergies. The present study aimed to elucidate the molecular mechanisms underlying HXS's therapeutic effects. Ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) was used to identify HXS components absorbed into the bloodstream. Network pharmacology, molecular docking, and molecular dynamics simulations were then conducted to explore the active ingredients and their regulatory mechanisms in fracture healing and blood stasis. Transdermal absorption tests identified 20 active compounds from HXS. Network pharmacology analyses using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform highlighted vanillic acid, demethyleneberberine, palmatine hydrochloride, luteolin, apigenin, and wogonin as key active ingredients. Molecular dynamics simulations further validated the stability, conformational changes, and interactions of these compounds with their target proteins. Analysis of the transdermal absorption samples revealed 291 potential active targets for HXS in treating fractures and blood stasis, of which 159 were common to both conditions. Protein-protein interaction (PPI) network analysis identified core targets including AKT1, ALB, EGFR, STAT3, and CTNNB1. Molecular docking confirmed strong binding interactions between HXS compounds and these core targets, while molecular dynamics simulations validated the stability and mechanistic plausibility of these interactions. This study provides a systematic elucidation of HXS's molecular mechanisms in fracture healing and blood stasis. Identification of active compounds, core targets, and their interactions offers a scientific basis for the therapeutic effects of HXS and supports the rational development of herbal-medicine-based interventions for fracture management and blood stasis treatment.

Identification and functional characterization of a regioselective O-methyltransferase involved in physcion biosynthesis in Polygonum cuspidatum.

Menopause increases cardiometabolic risk, partly by reducing the protective effects of estrogens and inducing gut microbiota dysbiosis, which can promote the production of atherogenic metabolites such as trimethylamine N-oxide (TMAO). Polyphenols may reduce TMAO levels, though interindividual variability limits reproducibility. We compared urinary and serum TMAO levels, and urinary trimethylamine (TMA) and dimethylamine (DMA) levels between healthy women of reproductive age (Pre-M, n = 120) and non-medicated postmenopausal women (Post-M, n = 90) using UPLC-QqQ-MS/MS. In Post-M women, we conducted a randomised, placebo-controlled crossover study to evaluate the effects of a polyphenol-rich extract mixture containing pomegranate, Polygonum cuspidatum, and red clover (sources of ellagitannins, resveratrol, and isoflavones) on TMAO, TMA, and DMA in the whole group and after metabotyping. Because medication is common in Post-M women due to age and cardiometabolic risk, trials in non-medicated participants are challenging, yet avoiding drug-diet interactions allows clearer attribution of dietary effects. Urinary TMAO and DMA levels were higher in Post-M than in Pre-M. No changes were observed in serum TMAO. However, the intervention reduced urinary TMAO and DMA versus baseline and placebo. The effects varied by metabotype. TMAO reduction was significant in urolithin A metabotype (UMA), equol producers (EP), and lunularin non-producers (LNP). Reductions and effect sizes were most pronounced in the metabotype clusters MC3 (UMA + EP + LP) and MC7 (UMA + EP + LNP), which represented 39% of participants. DMA decreased selectively in UMA. No correlations were found between TMAO or DMA changes and BMI, age at menopause onset, or years since menopause. These findings show that polyphenol supplementation reduces urinary TMAO in a metabotype-dependent manner and support metabotyping as a precision-health strategy to mitigate cardiometabolic risk after menopause.

Mechanism of Reynoutria Japonica Houtt and its active component luteolin in treating acute lung injury by targeting AKT1 to inhibit macrophage apoptosis and mitochondrial damage.

Polygonum cuspidatum alleviates hyperuricemia-induced renal injury via rectifying amino acid metabolism disorders.

Global lysine crotonylation profiling reveals metabolic and stress-responsive mechanisms in Reynoutria japonica.

Emodin glucosides, including emodins 1-O-glucoside (E1G), E6G, and E8G, possess significant pharmaceutical and nutraceutical potentials. Here, we identified PcUGT71BE11 from Polygonum cuspidatum as a bifunctional UGT capable of both glycosylating all three hydroxyl groups of emodin and deglycosylating E1G and E8G. Kinetic and time-course analyses showed that E1G was the first product, and its accumulation was pH-dependent. PcUGT71BE11 predominantly produced E1G at pH 8.8 but shifted to E6G at pH 6.8 due to decreased glycosylation and increased deglycosylation at acidic pH. The UDP-dependent deglycosylation was attenuated by sucrose synthase introduction, thereby enhancing E1G accumulation. Structural modeling, molecular dynamics, MM/GBSA binding free energy, and mutagenesis revealed the molecular basis: H19, D120, P190, and E395 governed the bifunctionality, while C144, I192, and L147 contributed to the regioselectivity. Notably, the L147N mutant enhanced the 8-O-glycosylation selectivity. This study elucidates the molecular mechanism of PcUGT71BE11 and provides a promising catalyst for producing diverse emodin glycosides.

Knotweed (Reynoutria japonica) is an invasive plant whose ecological dominance in its invasive range may partly rely on allelopathic interactions mediated by root exudates and especially secondary metabolites of wide polarity range. To elucidate the chemical diversity of these exudates, a metabolomic strategy combining liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) and gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS) was employed. After optimization of EcoRoot methodology with design of experiment (DoE), root exudates were sampled with 3 different sorption materials and analyzed using LC-ESI-MS/MS and GC-HRMS. Molecular networking was applied to MS/MS data to visualize chemical structural similarities and facilitate the identification of metabolites with a confidence annotation scale. Diverse families of compounds, including flavonoids, alkaloids, organic acids, terpenoids, and phenolics, were annotated and many of which could be implicated in allelopathic interactions or rhizospheric communication. Notably, molecular networks highlighted the presence of lineage-specific metabolites (Polygonaceae). Also, suspect screening was performed and led to confirmation of allelochemicals as root exudates: chlorogenic acid, catechin, and polydatin. GC-HRMS further revealed volatile organic compounds such as terpenoids or carbon disulfide, which may contribute to belowground chemical interference. These VOCs were trapped using another sorbent called Sorb-Star® and then thermodesorbed. This study underscores the value of integrated HRMS-based metabolomics for exploring the chemical diversity of R. japonica and provides candidate metabolites (isomers of selinene, carbon disulfide) for future studies in both fundamental and applied ecology and agronomy. Finally, this study presents the first metabolomic fingerprint of R. japonica grown in soil.

Protective effects of Polygonum cuspidatum bioactive constituents against cholestatic liver injury: A mechanistic review.

Development and mechanism of an XOD-targeting fermented traditional Chinese medicine formula for gout prevention in goslings.

Transcriptome insights reveal root hair inhibition and ROS imbalance in radish seedlings treated with rhizome extracts of invasive Fallopia species.

Objective: This study aimed to optimize the extraction of bioactive compounds from Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by evaluating the effect of ethanol concentration. Methods: Ultrasonic extraction was performed using ethanol concentrations of 0%, 30%, 50%, and 70%, and the resulting extracts were assessed for their chemical composition and multifunctional bioactivities. Results: The 70% Ethanol extract exhibited the highest total polyphenol and flavonoid contents and demonstrated the most potent antioxidant, enzyme-inhibitory, and antimicrobial activities, with significant differences (p < 0.05) compared to other concentrations. Chemical analysis identified tannic acid, emodin, and a variety of phenolic compounds as the primary bioactive constituents. Structural analyses using Field Emission Scanning Electron Microscopy (FE-SEM) and Fourier Transform Infrared (FT-IR) Spectroscopy revealed that 70% Ethanol induced the most pronounced structural changes to the cell wall, while FT-IR analysis confirmed the presence of O-H, C=O, C=C, and C-O functional groups, providing a mechanistic basis for the superior extraction efficiency and bioactivity. Conclusions: Ethanol concentration is a critical determinant for maximizing the bioactivity of P. cuspidatum. Extraction with 70% ethanol is identified as the optimal condition, supporting the potential of this plant as a source of natural bioactive compounds.

Polygonum cuspidatum is a well-known and versatile medicinal plant. In Taiwan, P. cuspidatum is typically found in central mountainous regions. Once acclimated, it can also thrive in flat areas, where it is known as flatland P. cuspidatum. Flatland P. cuspidatum has several advantages over alpine P. cuspidatum; for example, flatland P. cuspidatum grows faster and has larger leaves. This study enhanced the functionality of different parts of flatland P. cuspidatum (flowers, leaves, and rhizomes) by using microwave-assisted extraction (MAE) technology and Box-Behnken response surface methodology. Experiments revealed that the combination of MAE parameters that yielded optimal results was influenced by which plant part was used as input material. Regarding whitening activity, the extracts were ranked as follows: leaf > rhizome > flower. Leaf extracts had higher total flavonoid content, and rhizome extracts had higher total phenolic content. Regarding antiaging activity, the extracts were ranked as follows: rhizome > leaf > flower. The rankings for antimicrobial activity were as follows: leaf > rhizome > flower. Regarding anti-inflammatory activity, the extracts were ranked as follows: flower > rhizome > leaf. The rhizome extract exhibited slight cytotoxicity. UHPLC-UV-Q-TOF-HRMS/MS analysis identified 27, 34, and 37 bioactive compounds in the leaf, rhizome, and flower extracts, respectively. Given the relatively low pharmacological activity observed in the MAE-optimized flower extract, fermentation with Aspergillus oryzae was employed to enhance its efficacy. This process significantly enhanced the extract's pharmacological properties, including its whitening, antiaging, and antimicrobial properties. Increased levels of 3-O-caffeoylquinic acid, decursin, quercetin, quercitrin, kaempferol, resveratrol, epicatechin gallate, resveratrol-3-O-D-(2-galloyl)-glucopyranoside, resveratrol-4'-O-β-D-glucoside, apigenin, emodin-8-O-(6'-O-malonyl)-glucoside, physcion, emodin, and torachrysone in the fermented flower extract likely contributed to its enhanced pharmacological activities. The results of this study indicate that the newly developed flatland P. cuspidatum extracts can be considered viable substitutes for alpine P. cuspidatum extracts.

Understanding the distribution and diversity of invasive alien species is increasingly crucial to meet legal requirements and guide effective management. Portugal currently hosts a large number of invasive alien plant species, with significant environmental and socio-economic impacts. However, the distribution of most of these species across the territory and the factors driving their spread and diversity remain largely unexplored. To address this, we present the first atlas of invasive alien plants in mainland Portugal. A total of 96 terrestrial and aquatic invasive alien plants are presented, encompassing all species listed under Portuguese national legislation and the European Union’s list of invasive species of concern. Occurrence data were collected from a broad array of sources, including national and international biodiversity observation databases, citizen science data, literature, and data collections owned by managers and researchers—totaling approximately 85,000 records, with a mean of 879 records per species (range: 1–9,190). Each species was characterized based on multiple distributional parameters, and a k-means analysis was used to group species with similar distribution patterns. The richness of invasive plants was mapped at the municipality level. The drivers of their spatial variation were investigated using a comprehensive set of 30 variables representing multiple environmental and human factors. Results show that invasive plants are present in all mainland Portuguese municipalities but with high variability. Four main patterns of distribution were identified: species primarily located along the coast (e.g., Acacia saligna ) and a gradient of species with narrow (e.g., Reynoutria japonica ), moderate (e.g., Ipomoea indica ), and wide distribution ranges (e.g., Cortaderia selloana ). Invasive species richness was significantly higher in coastal and larger municipalities, particularly those closer to major urban centers and with a higher density of power lines. Our results provide the first comprehensive assessment of the distribution of invasive alien plants in mainland Portugal, establishing a much-needed baseline for future invasion prevention and management efforts.

Abstract Invasive plant species can significantly alter soil ecosystems, affecting biodiversity, nutrient cycling, and ecological balance. This study investigates the effects of Fallopia japonica and Solidago gigantea on soil nematode communities in riverbank and permanent grassland ecosystems across different geomorphological regions. It explores how these invasions affect soil properties and nematode community structures, with an experimental study testing the effects of organic matter from these two invasive plant species on soil health. The findings indicate that F. japonica in riparian ecosystems increased soil pH, reduced nematode abundance, especially among plant-parasitic nematodes. In the grassland, S. gigantea increased soil pH and bacterivore and fungivore populations while negatively affecting soil maturity and functional indices. Experimental trials demonstrated that organic matter from both invasive plants reduced nematode diversity and influenced soil moisture and pH. The study emphasises the need for effective environmental management strategies to mitigate the ecological impacts of invasive plant species on soil ecosystems. Information © The Authors 2025

Fungal-based soil food webs, not local plant nor abiotic properties, are most affected by rhizome-derived secondary metabolites of Japanese knotweed (Reynoutria japonica)

Background Polydatin, a natural component of Polygonum cuspidatum, exhibits potent anti-metabolic properties. The treatment with Poly (10 µm/L) effectively reversed the high glucose (HG)-induced reduction in acetylcholine (ACh)-elicited vasodilation in the aortas of Sprague-Dawley rats. Methods Male Sprague-Dawley rats were used to evaluate the effects of polydatin on endothelial function under HG conditions. Endothelium-dependent relaxation (EDR) was assessed in isolated thoracic aortic rings using ACh, with or without L-NAME or tempol. Human umbilical vein endothelial cells (HUVECs) were also treated under normal glucose (NG), HG, or HG + polydatin conditions. Gene expression (NLRP3, VCAM-1, GAPDH) was measured by RT-PCR, while protein levels of eNOS, iNOS, NLRP3, VCAM-1, and GAPDH were analyzed by western blotting. Results HG significantly impaired ACh-induced EDR in rat aortic rings, while polydatin (10 µmol/L) restored vascular responsiveness. Mechanistically, polydatin upregulated eNOS and suppressed iNOS expression, and its vasoprotective effects were partially inhibited by L-NAME, indicating nitric oxide (NO) pathway involvement. In both aortic tissues and HUVECs, HG markedly increased NLRP3 and VCAM-1 expression, which was effectively reversed by polydatin, indicating its anti-inflammatory action. Conclusion Polydatin counteracts hyperglycemia-induced endothelial dysfunction by enhancing eNOS-dependent NO signaling to restore vasodilatory capacity, while inhibiting NLRP3 inflammasome activation and downstream VCAM-1 expression to attenuate vascular inflammation. These dual mechanisms position polydatin as a therapeutic agent for preserving vascular function in diabeticconditions.

Ferroptosis, an iron- and reactive oxygen species (ROS)-dependent cell death process, plays a key role in acute kidney injury (AKI). Inhibiting ferroptosis has been shown to mitigate AKI severity, highlighting its potential as a therapeutic target. Lysine methyltransferase SETD2 regulates ferroptosis-related proteins and may serve as a novel target for ferroptosis inhibitors. Polydatin, a resveratrol glucoside derived from Polygonum cuspidatum , has demonstrated protective effects in septic-induced AKI, but its role in ischemia-reperfusion-induced AKI remains unclear. This study investigates whether Polydatin protects against ischemia-reperfusion-induced AKI in mice and explores the underlying mechanisms. A mouse AKI model was established by clamping renal pedicles for 30 min, followed by Polydatin (40, 80 mg/kg) administration 1 h before injury. Renal function was assessed via serum creatinine (SCr) and blood urea nitrogen (BUN) levels, alongside histopathological analyses. RNA sequencing identified potential mechanisms, and molecular docking examined Polydatin's interaction with SETD2. ChIP-qPCR was used to assess SETD2-mediated regulation of ACSL4 via H3K36me3 modification. SETD2 knockout mice were utilized to validate Polydatin's protective mechanism. Polydatin significantly improved renal function, reducing SCr, BUN, and injury biomarkers (NGAL, KIM-1). Invitro, it alleviated hypoxia/reoxygenation injury in HK-2 cells. Mechanistically, Polydatin inhibited ferroptosis by targeting SETD2, as confirmed by RNA sequencing and molecular docking. SETD2 overexpression negated Polydatin's benefits, while knockout enhanced protection. Polydatin protects against ischemia-reperfusion-induced AKI by inhibiting ferroptosis through SETD2 modulation, highlighting its potential for AKI treatment.

ABSTRACT Invasive plants pose a significant threat to ecosystems by disrupting the ecological balance, which includes the alteration of biogeochemical processes. Among the most aggressive invaders is Fallopia japonica, a species that thrives in riparian zones – critical interfaces between aquatic and terrestrial environments – where it significantly impacts biodiversity and ecosystem functions. Despite its success as an invader, the mechanisms that drive both the impact and success of F. japonica upon ecosystem processes remain poorly understood. Prior studies have suggested that F. japonica may exhibit traits such as a unique preference for ammonium over nitrate, potentially altering nitrogen availability for native plants like Urtica dioica. Additionally, it has been proposed that the species leverages phenolic compounds to influence soil biogeochemistry and nitrogen cycling. However, these processes lack comprehensive investigation. Using stable isotope labelling (15N and 13C), we found that, contrary to prior assumptions, F. japonica showed an overall lower uptake of both ammonium and nitrate relative to the native competitor, U. dioica. Although we expected a preference for ammonium, F. japonica instead exhibited a slight preference for nitrate. In addition, F. japonica demonstrated higher nitrogen-use efficiency and allocated more freshly assimilated carbon and nitrogen to root growth than U. dioica. These findings suggest that traits such as efficient nitrogen use and strategic root allocation may contribute to F. japonica's ability to establish itself in nitrogen-variable environments like riparian zones. By prioritising belowground biomass during early development, F. japonica may gain a competitive advantage that enables it to disrupt native plant communities and alter ecosystem dynamics. This study underscores the value of stable isotopes in understanding plant–soil interactions and informs strategies for managing invasive species in sensitive ecosystems.

The use of antioxidants in the dermatocosmetic industry has become increasingly popular to help protect and stabilize other sensitive active ingredients, prolonging the effectiveness and durability of the cosmetic product. Grape pomace, as the main by-product generated through winemaking, and Polygonum cuspidatum, concentrate bioactive metabolites with high antioxidant activity. Hydroalcoholic extracts obtained from grape pomace (Merlot and Feteasca Neagra varieties) and the root and flower of Japanese knotweed, respectively, alone and in mixtures, were characterized, and preliminary assays were conducted for their incorporation in two gel-based cosmetic formulations. The characterization of the extracts revealed the presence of catechin, vanillic acid, caffeic acid, myricetin, resveratrol, and kaempferol. The hydroalcoholic extract of P. cuspidatum flower and root was found to have the highest content of total phenolic compounds (10.920 ± 0.268 mg GAE/mL, respectively, 4.751 ± 0.072 mg GAE/mL), and the highest antioxidant activity (expressed as DPPH Radical Scavenging Capacity, IC50) by 28.04 ± 1.12 µg GAE/mL and 83.91 ± 1.13 µg GAE/mL, respectively. Catechin was the most abundant polyphenol found in pomace extract (687.87 mg/kg). The type and the concentration of the plant extract used in dermatocosmetic gel formulations influenced their antioxidant activity. Encapsulation of P. cuspidatum flower extract in liposomes prior to their incorporation into the gel formulation demonstrated the role of liposomes in enhancing the stability and modulation of phenolic compound delivery. It is worth noting that this dermatocosmetic formulation, which contains the flower extract of P. cuspidatum, was the subject of a pending patent application.