Glucosinolates (GSLs), one type of secondary metabolites mainly enriched in cruciferous plants, play important roles in environmental stress responses. However, the function and regulatory mechanism of GSLs in combating salt stress in broccoli are largely unclear. In this study, BolTGG1, a member of β-thioglucoside glucohydrolase (TGG) genes mainly functioning to hydrolyze GSLs, was identified in broccoli. BolTGG1 expression was upregulated under salt stress. Overexpression of BolTGG1 significantly improved the tolerance to salt stress in both broccoli and Arabidopsis. Subsequently, BolTGG1 was heterologously expressed in yeast and the BolTGG1 protein was isolated to evaluate its ability to hydrolyze GSLs. The results confirmed that exogenous addition of BolTGG1 could obviously increase the yield of sulforaphane (SFN), a major hydrolysis product of GSLs, in vitro. Meanwhile, the content of SFN was significantly increased in BolTGG1-overexpression transgenic broccoli plants. The role of SFN in the salt stress response was then explored. The results indicated that external application of SFN could directly enhance the salt resistance with the elevated activities of superoxide dismutase (SOD) and peroxidase (POD), and reduced malondialdehyde (MDA) levels in broccoli. Furthermore, BolMYB122 was confirmed to be the upstream regulatory transcription factor of BolTGG1. BolMYB122 positively regulated BolTGG1 transcription. In summary, these results indicated that BolMYB122-BolTGG1 module confers salt stress tolerance mainly by accelerating the generation of SFN. These findings provide new insights into the role and regulatory mechanism of GSL metabolites in plant salt stress response, and suggest a novel breeding strategy for stress resistance from the perspective of regulating plant secondary metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE:Kai-Xin-San (KXS) is a classic prescription for treating affective disorders in traditional Chinese medicine (TCM) over millennium. Although antidepressant effects of KXS have been demonstrated in both clinical and preclinical studies, antidepressant constituents and action mechanisms of KXS remain unclear. AIM OF THE STUDY:To reveal antidepressant constituents and action mechanisms of KXS using the method of "fraction-spectrum-effect-mechanism". MATERIALS AND METHODS:Homologous fractions of KXS were isolated through solvent extraction combined with macroporous resin chromatography methods. The quantification of the fractions was performed using chemical chromogenic methods. Antidepressant effects of the homologous fractions were assessed using tail suspension test (TST), forced swimming test (FST) and open field test (OFT) in mice. Two major fractions of total phenols (TP) and total saponins (TS) were chosen for further studies. The constituents of TP and TS were identified using UPLC-Q-Exactive Orbitrap-MS (UPLC-MS) method, and their antidepressant effects were verified on chronic restraint stress (CRS) model in mice by behavioral evaluations including sucrose preference test (SPT), TST, FST and OFT. Network pharmacology was used to predict antidepressant mechanisms of TP and TS based on the homologous constituents determined in UPLC-MS analysis. Proteomic analyses were undertaken with tissues of hippocampus (HP) and prefrontal cortex (PFC) in CRS mice treated by TP and TS. Western blot (WB) was employed to verify the findings in network pharmacology and proteomic analyses. RESULTS:Five homologous fractions of essential oils (EO), TP, TS, oligosaccharides (OL) and polysaccharides (PO) were isolated from KXS with homologous contents exceeding 50 %. Results of TST and FST indicated that all the five fractions significantly decreased the immobility time of mice, while OFT results exhibited that their locomotor activities remained unchanged. A total of 50 constituents and 114 constituents were identified in the TP and TS, respectively. CRS tests demonstrated that both TP and TS significantly increased the percent of sucrose consumption and decreased the immobility time in the TST and FST, without affecting their locomotor activities. Results of both network pharmacology and proteomic analyses primarily pointed to similar contents highly related to depression. GO analysis on TP and TS principally enriched in synaptic structures and functions, as well as neural metabolic and gene expression processes. KEGG analysis on TP and TS principally enriched in pathways of neurodegeneration-multiple diseases, as well as neurotrophin signaling pathway in HP and PFC. Results of WB verified that TP and TS significantly reversed the decrease in signature proteins of synaptic plasticity including presynaptic SYN1, postsynaptic PSD95 and GRIA1, as well as the underlying mechanism of BDNF/TrkB/Akt pathway in the HP and PFC induced by CRS. CONCLUSION:This is the first study on antidepressant constituents and action mechanisms of KXS guided by the method of "fraction-spectrum-effect-mechanism". Albeit the findings are still preliminary, it provides an example in methodology for unraveling the complicated chemical system and action mechanisms of TCM prescriptions.

Background: Meniere’s disease (MD) is a rare inner ear disorder characterized by endolymphatic hydrops and symptoms such as vertigo and hearing loss, with no curative treatment currently available. XuanYunNing tablets (XYN) have been clinically used to treat MD, but their molecular mechanisms remain unclear. Objective: This study aimed to systematically evaluate the pharmacological effects of XYN in a guinea pig model of MD and to elucidate the underlying molecular mechanisms of both MD pathogenesis and XYN intervention through integrated multi-omics analyses, including transcriptomics, proteomics, and bioinformatics. Methods: A guinea pig model of endolymphatic hydrops was induced by intraperitoneal injection of desmopressin acetate (dDAVP). Pharmacodynamic efficacy was evaluated via behavioral scoring and histopathological analysis. The differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) modulated by XYN treatment were identified using high-throughput transcriptomic and proteomic sequencing. These data were integrated through multi-omics bioinformatic analysis. Key molecular targets and signaling pathways were further validated using RT-qPCR and Western blotting. Results: Pharmacological evaluations showed that guinea pigs in the model group exhibited a 26% increase in endolymphatic hydrops area, while high-dose XYN treatment reduced this area by 19% and significantly improved functional parameters, including overall physiological condition (e.g., weight and general appearance), auricular reflexes to low-, medium-, and high-frequency sound stimuli, nystagmus, and the righting reflex. High-throughput sequencing combined with integrative omics analysis identified 513 potential molecular targets of XYN. Subsequent network and module analyses pinpointed the JAK-STAT signaling pathway as the central axis. Mendelian randomization (MR) analysis further supported a causal relationship between MD and metabolic, immune, and inflammatory traits, reinforcing the central role of JAK-STAT signaling in both MD progression and XYN-mediated intervention. Mechanistic studies confirmed that XYN downregulated IFNG, IFNGR1, JAK1, p-STAT3/STAT3, and AOX at both mRNA and protein levels, thereby inhibiting aberrant JAK-STAT pathway activation in MD model animals. In addition, a total of 125 chemical constituents were identified in XYN by UHPLC-MS analysis. ZBTB20 and other molecules were identified as potential blood-based biomarkers for MD. Conclusions: This study reveals that XYN alleviates MD symptoms by disrupting a pathological cycle driven by JAK-STAT signaling, inflammation, and metabolic dysfunction. These findings support the clinical potential of XYN in the treatment of Meniere’s disease and may inform the development of novel therapeutic strategies.

The curd is a unique indeterminate inflorescence structure, which is the most important edible organ of broccoli. Premature bolting of the curd is a major factor that limits the yield and marketability of broccoli. However, the formation and development of broccoli curd, especially the bolting of curd, remain poorly understood. In this study, BolAG, a homologous gene of the floral development gene AtAG, was highly expressed in curd and floral organ of broccoli. Overexpression of BolAG in broccoli significantly promoted curd development, leading to early bolting. Correspondingly, RNA interference (RNAi) of BolAG resulted in arrested curd development, leading to delayed or even absent bolting. Physiological analyses indicated that BolAG-overexpressing broccoli exhibited higher starch and soluble sugar contents (SSC) compared to the wild-type (WT). RNA-Seq analysis also identified significant enrichment of genes related to starch metabolism. Among them, the starch hydrolysis-related gene BolBAM4 was confirmed to be the target gene of BolAG. Additionally, BolAG was confirmed to interact with ribose-5-phosphate isomerase 2 (BolRPI2), a protein involved in the pentose phosphate pathway and their interaction enhanced the transcriptional promotion of BolAG on BolBAM4, accelerating starch degradation and thereby leading to the accumulation of soluble sugars. While BolRPI2 was demonstrated to have a positive effect on chloroplast function and promote starch synthesis, BolAG appeared to promote starch synthesis in a BolRPI2-independent manner. These results confirm that the BolAG-BolRPI2-BolBAM4 module plays a crucial role in regulating starch metabolism, providing a theoretical foundation for addressing industrial losses due to premature bolting.

OBJECTIVE: To analyze the chemical components of Mudan Huaban recipe ((sic)(sic)(sic)(sic)(sic), MHR) and evaluate its efficacy and possible mechanism in melasma mice. METHODS: The chemical compositions of MHR were determined by the ultra-high performance liquid chromatography coupled with quadrupole-exactive mass spectrometry method. Female C57BL/6 mice were exposed to ultraviolet B and progesterone for 21 d to induce melasma, and Fontana-Masson staining was used to assess the effects of MHR on melasma. Luteinizing hormone (LH), estradiol, and follicle-stimulating hormone levels were detected by enzyme-linked immunosorbent assay. The superoxide dismutase (SOD) activity and malonic dialdehyde content were detected by chemiluminescence. Tyrosinase and related proteins expressions were detected by Western blots and immunohistochemistry. RESULTS: A total of 43 chemical components were identified in MHR, MHR significantly decreased the melanin particles of melasma mice. MHR treatment significantly reversed the high contents of LH and low activity of SOD in models. MHR significantly reduced the higher levels of tyrosinase, tyrosinase-related proteins-1 (TRP-1), TRP-2, microphthalmia-associated transcription factor and phosphorylation of cyclic adenosine monophosphate response element-binding protein (p-CREB)/CREB in the skin of melasma mice. CONCLUSIONS: MHR protects against melasma via regulation of sex hormones, oxidative stress, and melanogenesis-related proteins, suggesting its possible use as a supplement and alternative drug therapy for melasma. (c) 2025 JTCM. All rights reserved.

ABSTRACTPanax notoginseng (P. notoginseng) is one of the most famous natural medicines and widely used to promote blood circulation in health care. However, the active component group of P. notoginseng for activating blood is not clear. We aim to screen and validate the pharmacodynamic component group (PCG), which could exert the same blood‐activating effect as P. notoginseng. To clarify the active components, the chemical components were determined by liquid chromatography‐tandem mass spectrometry, and the fingerprint of P. notoginseng was established. Twenty candidate active monomers were selected through the spectrum–effect relationship analysis. Eleven active monomers, including Ginsenoside Rg1, Rb1, Rd, F1, Rh1, Rg2, Rb2, Rg3, and Rk1 and Notoginsenoside R1 and R2, were screened out as the PCG through validation by platelet aggregation test. Among them, the antiplatelet aggregation activity of Ginsenoside Rh1 was directly confirmed for the first time. The active component group could exert similar efficacy to the P. notoginseng extract in vitro and in vivo through the validation of in vitro platelet aggregation test and the rats with cerebral ischemia. This study laid the foundation for the quality evaluation of P. notoginseng and provided a reference for the research on the material basis of the pharmacodynamics of other Chinese herbs.

Modulating cell endocytosis activity to reduce host susceptibility to virus represents a promising strategy for antiviral drug development. In this study, we reveal that lactate transporter SLC16A3 is a critical host factor for reducing diverse virus invasion. By performing metabolomics, proteomics, and thermal proteome profiling experiments, AP1G1, a pivotal protein involved in cellular endocytosis, was indiscriminately screened as a chaperone of SLC16A3. SLC16A3 decides the membrane enrichment of AP1G1 by protein interaction, thereby influencing host susceptibility to diverse viruses. This conclusion was further validated in SLC16A3 knocked-down cells, which indicated a broad-spectrum target for anti-virus drug development to recede virus entry by blocking the interaction between AP1G1 and SLC16A3. This conclusion has been validated with a patent medicine Shufengjiedu (SFJD). SFJD exhibits strong effect in decreasing the susceptibility of host cells to viral infections. Molecularly, SFJD administration results in disrupting the interaction between SLC16A3 and AP1G1 and reduced membrane localization of AP1G1. Consequently, it achieves a receded endocytosis activity of host cells on viral particles. This provides evidence for the practicability of the SLC16A3-AP1G1 strategy. Taken together, the regulation on the SLC16A3-AP1G1 interaction represents a broad-spectrum and practicable antiviral strategy. This study offers insights into a novel approach for inhibiting viral infections through the alteration of host susceptibility and advances the idea for antiviral drug development.IMPORTANCEWe have discovered that a broad-spectrum antiviral strategy, highlighting the lactate transporter SLC16A3 as a critical determinant of host cell susceptibility to viruses. SLC16A3 was found to interact with AP1G1, which is a pivotal protein involved in cellular endocytosis. Disrupting the interaction between AP1G1 and SLC16A3 leads to reduced membrane localization of AP1G1, thereby reducing the host cell endocytosis of viral particles. Importantly, we found that the patent medicine Shufengjiedu (SFJD can significantly reduce the susceptibility of host cells to viral infection through this mechanism, providing evidence for the practicability of SLC16A3-AP1G1 strategy. Taken together, the modulation on the SLC16A3-AP1G1 interaction represents a broad-spectrum and practicable antiviral mechanism. This study offers novel insights into strategies for inhibiting viral infections through the alteration of host susceptibility and advances the idea for antiviral drug development.

Natural products (NPs) make a major contribution to drug development, offering a huge molecule pool for drug leads. Nevertheless, the pharmaceutical industry and academy have declined their enthusiasm to NPs research since the great challenges in elucidating the complex component and intricate mechanism of NPs. Here, we introduce an efficient fragment-based target research (FBTR) approach for pharmacology study and optimization of NPs. Focusing on the core fragment within the molecules of NPs, we screen the outstanding activity that be triggered, and corresponding target. Finally, drug optimization was carried out around the molecules that obtaining the activity-related core fragment and verified both in vitro and in vivo. With this approach, we obtained an optimized NPs named Erigeron breviscapus polyphenols (EBP) with definite target. After optimization, EBP plus (EBPP) not only trigger immunogenic cell death (ICD) of glioblastoma (GBM) cells effectively by targeting to Cys105 amino acid site of Fas-associating protein with a novel death domain (FADD) protein, but also prolong the survival of GBM mice by an average of 17.6 days. Significantly, our investigation presents an approach for addressing challenges in NPs development and opening up new opportunities for drug discovery. Our findings demonstrate the utility of FBTR in exploring the function of NPs, revealing the target, and advancing drug optimization for stronger clinical translation.

Background:Pseudobulbus Cremastrae seu Pleiones (Shancigu), a traditional Chinese medicine (TCM), has been extensively used in clinical practice for the treatment of various tumors, particularly liver cancer. Shancigu is classified into two commercial specifications-"Maocigu" and "Bingqiuzi"-which exhibit significant differences in appearance, chemical composition, and price, posing challenges for the quality control of medicinal materials. Purpose:The aim of this study was to clarify the quality evaluation indicators based on the anti-liver cancer active components in Shancigu and to establish a reliable quality evaluation method to preliminarily assess the quality of Shancigu from different commercial specifications and production areas. Methods:Twenty-six batches of Shancigu samples were collected. High-performance liquid chromatography (HPLC) was used to establish fingerprint spectra. In vitro anti-liver cancer pharmacological effect indicators were analyzed using the CCK-8 assay and scratch wound healing assays. Through spectrum-effect relationship analysis, serum pharmacochemistry analysis, and in vitro/in vivo anti-liver cancer activity evaluation, the effective component combinations of Bingqiuzi and Maocigu were identified and validated. Gray relational analysis (GRA) and the technique for order preference by similarity to ideal solution (TOPSIS) were subsequently applied to assess the quality of Shancigu based on their different specifications and origins. Results:Eleven key anti-liver cancer active components from Shancigu were screened and confirmed, namely, malic acid, citric acid, 2-isobutylmalic acid, gastrodin, batatasin III, 2-p-hydroxybenzyl-5,3'-dihydroxy-3-methoxybibenzyl, coelonin, 1-p-hydroxybenzyl-2,7-dihydroxy-4-methoxyphenanthrene, blestriarene A, blestriarene B, and monbarbatain A. These components are present in Bingqiuzi and Maocigu in different proportions, and the anti-liver cancer pharmacological effects of the effective component combinations were found to be equivalent to those of the original materials, both in vitro and in vivo. These 11 components can be used as indicators for evaluating the quality of Shancigu. Quality evaluations revealed no significant differences between Bingqiuzi and Maocigu. For Bingqiuzi, medicinal materials produced in Guizhou and Yunnan were of better quality; for Maocigu, those from Guizhou and Sichuan were superior. Conclusion:In this study, we established quality evaluation criteria for Shancigu and developed an innovative method to comprehensively assess the quality of Shancigu from different commercial specifications and production regions. By integrating component analysis with anti-liver cancer activity assessment, this research provides a valuable reference for the quality evaluation of other Chinese medicinal materials.

OBJECTIVE:To analyze the chemical components of Mudan Huaban recipe (, MHR) and evaluate its efficacy and possible mechanism in melasma mice. METHODS:The chemical compositions of MHR were determined by the ultra-high performance liquid chromatography coupled with quadrupole-exactive mass spectrometry method. Female C57BL/6 mice were exposed to ultraviolet B and progesterone for 21 d to induce melasma, and Fontana-Masson staining was used to assess the effects of MHR on melasma. Luteinizing hormone (LH), estradiol, and follicle-stimulating hormone levels were detected by enzyme-linked immunosorbent assay. The superoxide dismutase (SOD) activity and malonic dialdehyde content were detected by chemiluminescence. Tyrosinase and related proteins expressions were detected by Western blots and immunohistochemistry. RESULTS:A total of 43 chemical components were identified in MHR, MHR significantly decreased the melanin particles of melasma mice. MHR treatment significantly reversed the high contents of LH and low activity of SOD in models. MHR significantly reduced the higher levels of tyrosinase, tyrosinase-related proteins-1 (TRP-1), TRP-2, microphthalmia-associated transcription factor and phosphorylation of cyclic adenosine monophosphate response element-binding protein (p-CREB)/CREB in the skin of melasma mice. CONCLUSIONS:MHR protects against melasma via regulation of sex hormones, oxidative stress, and melanogenesis-related proteins, suggesting its possible use as a supplement and alternative drug therapy for melasma.

Glucosinolates (GSLs) are secondary metabolites popularly existing in Brassicaceae. However, the role and regulation of GSLs in environmental stress response remain ambiguous. Here, Methylthioalkylmalate synthase 1 (BolMAM1), a GSL biosynthesis-associated gene, was identified in broccoli. The overexpression of BolMAM1 enhanced both salt and drought tolerance in broccoli, along with an increase in aliphatic GSLs, such as 2(R)-hydroxy-3-butenyl GSL (progoitrin). Progoitrin could induce ABA accumulation in vitro. Consistently, ABA homoeostasis was modulated to accumulate more ABA in OEX-BolMAM1 transgenic broccoli. Moreover, sulforaphane (SFN), one of the hydrolysates of aliphatic GSLs, also exhibited accelerated accumulation in OEX-BolMAM1 transgenic broccoli. External application of SFN could rapidly induce stomatal closure. Furthermore, BolMYB28 was demonstrated to directly bind to the promoter of BolMAM1 and activate its transcription. These results indicated that BolMYB28-BolMAM1 confers tolerance to salt and drought stresses mainly by accelerating the biosynthesis of certain specific aliphatic GSLs. Progoitrin is a newly reported GSL positively regulating ABA accumulation. SFN may act as a signaling molecule to regulate stomatal behavior. These findings reveal the role of BolMYB28-BolMAM1 in abiotic stress response via regulating aliphatic GSL biosynthesis, and suggest the applications of progoitrin and SFN as natural defence factors to resist abiotic stresses in plants.

Connective tissue disease-associated interstitial lung disease (CTD-ILD) is a systemic autoimmune disease with high morbidity and hazard, characterized by progressive pulmonary inflammation and fibrosis. The monomer formulation of polydatin and curcumin (PD + Cur) for lung injury in CTD-ILD was optimized from Curcumae Longae Rhizoma (Curcuma Longa L.) and Polygoni Cuspidati Rhizoma Et Radix (Polygonum cuspidatum Sieb. et Zucc.). Mice with CTD-ILD-like lung injury were established by a single intratracheal drip of bleomycin. After intervening in model mice for 4 weeks, PD + Cur attenuated alveolar atrophy, fibrillar collagen formation, and thickened alveolar septa in the lung, improved serum biomarkers TOLLIP, MUC5B, KL-6, SP-D, and RCN3, and suppressed serum immunoinflammatory factors IL-6, CCL-18, and SF. The transcriptome sequencing showed that PD + Cur ameliorated CTD-ILD mainly by regulating aberrant immunoinflammation, which was further confirmed by proteomics that the PI3K/AKT/TGF-β pathway was a key pathway. Further, PD + Cur was found to affect amino acid metabolism in the serum significantly. The B-type receptor for GABA (GABBR) agonist baclofen was further found to attenuate CTD-ILD-like lung injury and modulate PI3K/AKT/TGF-β signaling. However, the inhibition of AKT, transforming growth factor beta receptor type 3 (TGFβR3), a key indicator downstream of PI3-kinase subunit p85-alpha (PI3KR1), by PD + Cur was reversed after intervention with the GABBR receptor inhibitor CGP52432. PD + Cur has an ameliorative effect on CTD-ILD-like lung injury by targeting GABBR to modulate the PI3K/AKT/TGF-β pathway.

Traditional antiviral strategies primarily rely on vaccines and virus protein-targeting drugs, which adopt a virus-targeting approach. However, the rapid mutation of viruses often leads to vaccine failure and drug resistance, highlighting the limitations of these conventional methods. Consequently, the development of novel broad-spectrum, host-targeting antiviral strategies has become a major research focus. Itaconate, an endogenous immunomodulatory metabolite, inhibits viral replication via post-translational modifications; however, its mechanism in suppressing viral endocytosis remains unclear. This study demonstrates that itaconate inhibits viral endocytosis by covalently modifying the Cys128 site of the adaptor-related protein complex 1 gamma 1 subunit (AP1G1), thereby providing a new target for host-directed antiviral drug development. It was found that itaconate binds to AP1G1 at Cys128, impairing its interaction with clathrin, which inhibits clathrin-mediated viral particle uptake and reduces cellular susceptibility to infection (i.e., the likelihood of cells being infected by viruses and undergoing infection). Furthermore, the natural product Licochalcone B was identified as targeting the same site as itaconate. In both BEAS-2B cell models and mouse infection models, Licochalcone B reduced pulmonary viral loads by over 95%. This study is the first to propose and validate the feasibility of inhibiting broad-spectrum viral infection by targeting AP1G1, elucidating a novel molecular mechanism of itaconate-mediated regulation, offering a new target for broad-spectrum antiviral drug development, and identifying Licochalcone B as a promising broad-spectrum antiviral agent.

Larix kaempferi, one of the representative gymnosperms, is the main afforestation and timber species in the world. As one essential micronutrient, zinc plays a vital role in Larix kaempferi growth, yet its overaccumulation causes substantial growth inhibition. However, the underlying mechanism of zinc stress response in Larix kaempferi is still not well elucidated. In the present study, a potential zinc stress-related gene, LkCNR6 (Cell number regulator 6) was identified in Larix kaempferi. LkCNR6 was confirmed to localize in the plasma membrane and displayed rapidly inducible expression patterns under zinc stress. To uncover the roles of LkCNR6, the LkCNR6-overexpressing transgenic Arabidopsis lines were created. The phenotypic data confirmed that ectopic overexpression of LkCNR6 significantly improved zinc stress tolerance in Arabidopsis, although the LkCNR6-overexpressing transgenic plants displayed slower growth rate and smaller aerial organ size than that of the wild-type plants. Zinc accumulation assay indicated that the improved tolerance to zinc stress was mostly attributed to increase zinc excretion in the overexpressing LkCNR6 transgenic plants. Furthermore, yeast two-hybrid assay identified that LkHMP1, LkWAKL4 and LkRTFL, were physically interacted with LkCNR6. The interaction between LkCNR6 and LkHMP1, a protein associated with metal transport, was further confirmed by bimolecular fluorescence complementation (BiFC) assay. In summary, these results demonstrated that LkCNR6 plays important roles in positively responding zinc stress and negatively regulating organ development. These findings provide new insights into the regulatory mechanism of LkCNR6-mediated zinc stress response, and suggest the application of LkCNR6 in breeding high-heavy metal resistance crops. LkCNR6 from Larix kaempferi confers zinc stress tolerance mostly by increasing zinc excretion from the plant body but negatively regulating growth and development in Arabidopsis.

OBJECTIVES:This study was designed to investigate the pharmacological activity and therapeutic mechanism of Mahuang Xixin Fuzi decoction (MXFD) on migraine.METHODS:Migraine model rats induced by nitroglycerin were established, and then orally administered with MXFD for 7 days. Blood and urine samples were collected to identify differential metabolites with metabolomics. To integrate the findings from network pharmacology and metabolomics analysis, the metabolites and targets related to MXFD therapy for migraine were filtered.KEY FINDINGS:MXFD was found to alleviate the symptoms of migraines in rats. After treatment with MXFD, nine metabolites were found to be regulated and returned to normal levels. MXFD acted directly on nine key targets including MAOB, MAOA, ADRB1, ADRB2, ADRB3, ADORA2A, ADORA2B, DRD5, and HTR4 and regulated two out of nine metabolites, namely deoxycholic acid and 5-methoxyindoleacetate.CONCLUSIONS:The study found that MXFD can alleviate migraines through multitarget and multicomponent interaction networks.

Broccoli, a cruciferous vegetable, has a unique indeterminate inflorescence structure known as curds. It is the main edible organ of broccoli and has a rich nutritional value and health benefits. However, the formation and development mechanism of the curd is still not well understood. In the present study, the shoot apical meristem (SAM) stage and three different development stages of curd (formation stage (FS), expansion stage (ES), and maturation stage (MS)) were identified and subjected to transcriptome sequencing to uncover the potential genes and regulatory networks involved in curd formation and development. The results indicated that the genes associated with the development of SAM such as BolAP1A, BolAP1C, BolCAL, and BolAGL6 play an important role in the abnormal differentiation of the curd apical buds. The genes, BolFRI, BolbHLH89, BolKAN4, BolAGL12, and BolAGL24, displayed significantly differential expression patterns in curd development may function in the regulation of the transition from inflorescence meristem (IM) to floral meristem (FM). Moreover, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed genes (DEGs) indicate that phytohormones, such as auxin (AUX), gibberellins (GA), and abscisic acid (ABA) also play an important role in SAM proliferation and the transition from SAM to IM. In addition, the genes regulating photosynthetic reaction (BolLHCA1, BolLHCB1, BolPsbO, etc.) have a key involvement in the differentiation of secondary IMs during curd expansion. The genes associated with the metabolism of starch and sucrose (e.g., BolSPS4, BolBAM4) were significantly upregulated at the MS should contribute to the maturation of the curd. These findings provide new insights into the potential key regulatory factors and metabolic pathways involved in the formation and development of broccoli curds.

Airway remodelling in lung diseases can be treated by inhibiting excessive smooth muscle cell proliferation. Zedoarondiol (Zed) is a natural compound isolated from the Chinese herb Curcuma longa. The caveolin-1 (CAV-1) is widely expressed in lung cells and plays a key role in platelet-derived growth factor (PDGF) signalling and cell proliferation. This study aims to investigate the effect of Zed on human bronchial smooth muscle cell (HBSMC) proliferation and explore its potential molecular mechanisms. We assessed the effect of Zed on the proliferation of PDGF-stimulated HBSMCs and performed proteomic analysis to identify potential molecular targets and pathways. CAV1 siRNA was used to validate our findings in vitro. In PDGF-stimulated HBSMCs, Zed significantly inhibited excessive proliferation of HBSMCs. Proteomic analysis of zedoarondiol-treated HBSMCs revealed significant enrichment of differentially expressed proteins in cell proliferation-related pathways and biological processes. Zed inhibition of HBSMC proliferation was associated with upregulation of CAV1, regulation of the CAV-1/PDGF pathway and inhibition of MAPK and PI3K/AKT signalling pathway activation. Treatment of HBSMCs with CAV1 siRNA partly reversed the inhibitory effect of Zed on HBSMC proliferation. Thus, this study reveals that zedoarondiol potently inhibits HBSMC proliferation by upregulating CAV-1 expression, highlighting its potential value in airway remodelling and related diseases.

Due to the multitudinous structural types of glycosylated com-ponents,accurate identification of glycosylation modifications and secondary metabolite structures in herbs remains a challenge for natural drug analysis and new drug discovery[1].

Sulforaphane (SFN) is one of the hydrolysates of glucosinolates (GSLs), primarily derived from Brassica vegetables like broccoli. In clinical therapy, SFN has been proven to display antimicrobial, anticancer, antioxidant, and anti-inflammatory properties. However, the antimicrobial effects and mechanism of SFN against plant pathogens need to be further elucidated, which limits its application in agriculture. In this study, the genetic factors involved in SFN biosynthesis in 33 B. oleracea varieties were explored. The finding showed that besides the genetic background of different B. oleracea varieties, myrosinase and ESP genes play important roles in affecting SFN content. Subsequently, the molecular identification cards of these 33 B. oleracea varieties were constructed to rapidly assess their SFN biosynthetic ability. Furthermore, an optimized protocol for SFN extraction using low-cost broccoli curds was established, yielding SFN-enriched extracts (SFN-ee) containing up to 628.44 μg/g DW of SFN. The antimicrobial activity assay confirmed that SFN-ee obtained here remarkably inhibit the proliferation of nine tested microorganisms including four plant pathogens by destroying their membrane integrity. Additionally, the data demonstrated that exogenous application of SFN-ee could also induce ROS accumulation in broccoli leaves. These results indicated that SFN-ee should play a dual role in defense against plant pathogens by directly killing pathogenic cells and activating the ROS signaling pathway. These findings provide new evidence for the antimicrobial effect and mechanism of SFN against plant pathogens, and suggest that SFN-ee can be used as a natural plant antimicrobial agent for crop protection and food preservation.

Ethnopharmacological relevanceThe traditional Chinese medicine (TCM) formula Banxia Xiexin decoction (BXD) has definite therapeutic effect in treating stress-induced gastric ulceration (SIGU) and many other gastrointestinal diseases, but its effect on gastric lymphatic pumping (GLP) remains unclear.Aim of the studyElucidating the role of GLP in SIGU and BXD treatment, and exploring the molecular mechanisms of GLP regulation.Materials and methodsIn vivo GLP imaging were performed on SIGU rat model, and the lymphatic dynamic parameters were evaluated. Gastric antrum tissues and serum were collected for macroscopic, histopathological and ulcerative parameters analysis. Gastric lymphatic vessel (GLV) tissues were collected for RNA-Seq assays. Differentially expressed genes (DEGs) were screened from RNA-Seq result and submitted for transcriptomic analysis. Key DEGs and their derivative proteins were measured by qRT-PCR and WB.ResultsGLP was significantly suppressed in SIGU rats. BXD could recover GLP, ameliorate stomach lymphostasis, and alleviate the ulcerative damage. Transcriptome analysis of GLV showed the top up-DEGs were concentrated in smooth muscle contraction signaling pathway, while the top the down-DEGs were concentrated in energy metabolism pathways especially fatty acid degradation pathway, which indicated BXD can promote lymphatic smooth muscle contraction, regulate energy metabolism, and reduce fatty acid degradation. The most possible target of these mechanisms was the lymphatic smooth muscle cells (LSMCs) which drove the GLP. This speculation was further validated by the qRT-PCR and WB assessments for the level of key genes and proteins.ConclusionsBy activating the smooth muscle contraction signaling pathway, restoring energy supply, modulating energy metabolism program and reducing fatty acid degradation, BXD effectively recovered GLP, mitigated the accumulation of inflammatory cytokines and metabolic wastes in the stomach, which importantly contributes to its efficacy in treating SIGU.