ETHNOPHARMACOLOGICAL RELEVANCE:Parameria laevigata has traditionally been used for treatment of traumas caused by falls, fractures, contusions and strains, but its efficacy in wound healing related to the traditional use has not yet been supported by experiments. AIM OF THE STUDY:To validate the effects of P. laevigata in diabetic wound healing, and explore bioactive components and potential pathway. MATERIALS AND METHODS:The effects of P. laevigata on wound healing were evaluated using diabetic mice. Bioactive compounds were obtained by bio-guided isolation, and their bioactivities were evaluated using cellular models. Metabolomics, network pharmacology and molecular docking were applied to explore pathway, which were further validated by reverse transcription-quantitative polymerase chain reaction. RESULTS:P. laevigata enhanced wound healing of diabetic mice roughly equal to bFGF by reducing inflammation and oxidative stress. It reduced O-acetyl-L-serine, and increased 2-pyrocatechuic acid and 5-methylcytidine metabolite levels of skin tissues. Moreover, it downregulated Tnf-α, Il-1β, Il-6 and Nf-κb p65 mRNA, and also upregulated Il-10 mRNA, likely contributing to the modulation of PI3K/AKT/MAPK signaling pathway. In addition, four bioactive compounds were isolated, which might protect endothelial cells from lipopolysaccharide-induced damage and enhance angiogenesis by binding proteins such as IRS, PI3K, AKT, RAS, ERK, TNF-α, IL-1β, IL-6, IL-10 and NF-κB. CONCLUSIONS:P. laevigata with four compounds exhibited bioactivity on wound healing, might involving the PI3K/AKT/MAPK signaling pathway.

Eight previously undescribed benzylisoquinoline alkaloid dimers, neothalfines A-H (1-8), alongside thirteen known analogs (9-21), were isolated from roots of Thalictrum wangii B. Boivin. The structures and absolute configurations were elucidated based on extensive spectroscopic data (HRESIMS, NMR, and IR) in conjunction with TDDFT-ECD calculations. Evaluation of all identified alkaloids revealed that twelve dimers inhibited marked anti-proliferative activity against three human cancer cell lines (A549, MHCC97H, and U-87MG) with IC50 values ranging from 2.1 to 38.1 μM. Among them, dimer 8 was particularly potent, displaying IC50 values of 3.5, 2.1, and 2.8 μM against the respective cell lines, outperforming the positive control. Mechanistic studies revealed that 8 induces mitochondrial dysfunction, ROS-mediated apoptosis, and inhibits colony formation. Network pharmacology and molecular docking identified SRC, EGFR, and GSK3β as core targets, further supporting its role in mitochondrial energy metabolism dysregulation.

Plants of Salvia are frequently used for treatment infections of boils and sores in Traditional Chinese medicine, which might inhibit microbial. In this work, the chemical constituents of the effective fractions were analyzed using UPLC-Q-TOF-MS, and multivariate analysis and network pharmacology were used to predict potential antibacterial substances and their mechanism. Moreover, against methicillin-resistant Staphylococcus aureus (MRSA) effect and pathway of bioactive compounds were validated by experiments in vitro and in vivo. Two unreported anti-MRSA compounds, miltirone and przewaquinone A were proposed and then validated by serial experiments. They acted as bactericides and biofilm scavengers, respectively, and both targeted on the cell membrane, causing leakage of contents and affecting intracellular metabolism, leading to bacterial death without significant toxicity. Furthermore, miltirone ameliorated skin wound infection caused by MRSA in mice, roughly equal to vancomycin. The research supported the traditional use of Salvia plants, and presented two potent bactericidal agents for further investigation.

ETHNOPHARMACOLOGICAL RELEVANCE:Dendrobium, commonly known as "Shihu", are the well-known Chinese traditional medicine with the ability of strengthening "Yin", tonify five viscera, lighten the body, prolong life span that were involved in immunity regulation. However, their T-cell-mediated cellular immune function was kept unknown. AIM OF THE STUDY:To investigate its T-cell-mediated cellular immune function, and reveal bioactive constituents and mechanism. MATERIALS AND METHODS:The eliminating of mutant cells (cancer cells) by T-cell-mediated cellular immune was used to evaluate T-cell immunomodulation function through the non-traditional cytotoxicity T-cell immunomodulatory anti-tumor model combination with bioactivity-guided isolation. The LDH killing and ELISA analysis were used to examine killing efficacy and cytokines. The vaccine adjuvant model of mouse breast cancer was used to evaluate T-cell immunomodulation efficacy in vivo. The proteome and phosphoproteome analysis were used to mechanism research combined with blocking rescue analysis. RESULTS:"Shihu" fraction (YD198-6) showed the T-cell immunomodulation killing cancer cells bioactivity among the seven extracts, and fifteen known compounds were identified by bioactivity-guided isolation, which revealed the compound 3,4',5-trihydroxy-3'-methoxybiphenyl (THMB) was a better bioactive constituent. Further mechanism research suggested THMB regulated T-cell against multiple tumor cells major through enhancing JAK3-STAT1 signal to upregulate granzyme B, perforin, IL-1β. Moreover, in mice TNBC model, the primary tumor growth was significantly inhibited about 62%, and lung metastasis was reduced about 51% accompanied with significantly improving overall survival (54 days vs 37 days, p < 0.05). CONCLUSION:These results demonstrated that "Shihu" could enhance T-cell mediated cellular immune function to protect health, and revealed its bioactive constituent and mechanism.

Ni-Fe bimetallic catalysts demonstrate high catalytic activity and strong resistance to deactivation during biomass steam reforming (BSR). However, their activation and reaction with steam (H2O) are complex, and a deeper understanding of the underlying mechanisms is crucial for enhancing catalyst stability. This study systematically investigates the influence of H2O on the deactivation mechanisms of Ni-Fe/Al2O3 catalysts, focusing on its dual role in modulating carbon deposition patterns and triggering metal particle sintering. Our experimental results reveal a paradoxical effect of steam: the introduction of H2O reduces the total amount of carbon deposits, it promotes the formation of structurally disordered carbon. Crucially, this disordered carbon is identified as the primary driver of catalyst deactivation. It not only physically blocks active sites but also accelerates the sintering of metal particles, leading to profound alterations of the catalyst’s surface architecture and a consequent loss of performance. Combining experimental results and DFT calculations, we reveal the energy barriers for carbon evolution and correlate them with catalytic stability. These insights provide a theoretical foundation for improving the deactivation resistance of the catalyst, controlling the structural type of carbon deposits, and optimizing the anti-sintering performance of the etallic catalysts.

Macadamia nutshell, an abundant agro-industrial byproduct, afforded 5 new and 25 known compounds in our investigation, in which macyclophanes A-C feature rare 5-alkylresorcinol-derived 6/18/6 macrocyclic framework reported for the first time. Moreover, new compound macyclophane A showed the most potent bioactivity against Rhizopus stolonifer, with a MIC value of 16 μg/mL, and was associated with disruption of plasma membrane integrity and leakage of cellular contents. Furthermore, macyclophane A suppressed R. stolonifer infection of strawberry effectively, comparable to myclobutanil. The findings suggest that macadamia nutshells might be a promising sustainable natural source for postharvest disease control.

Alstolactines L-N (1-3), three rare monoterpenoid indole alkaloids with complex condensed rings, were isolated from Alstonia scholaris. Among them, 2 possesses two unique bridging oxygen atoms forming a highly fused 6/5/ 6/6/6/6 ring system. The structures of the isolates were elucidated by spectroscopic analyses and electronic circular dichroism calculations. Furthermore, all compounds exhibited anti-inflammatory bioactivity in LPSinduced RAW264.7 cells.

Background The macromolecular monoclonal antibodies targeting CTLA-4/PD-1/PD-L1 have revolutionized cancer immunotherapy. However, their clinical efficacy is limited by poor tissue penetration, strong immunogenicity and frequent cytokine release syndrome (CRS) risk. In contrast, natural small molecules can effectively overcome these challenges due to their strong penetration, minimal immunogenicity and low CRS risk. Purpose Developing natural small molecule-mediated immunotherapeutic drugs represents a promising strategy for improving current cancer immunotherapy. Study design A non-traditional T-cell immunomodulatory anti-tumor model was utilized to explore these potential molecules. Methods LDH release assays and ELISA were used to assess T-cell immune killing activity against tumor cells and cytokine production. The mouse colon cancer lung metastasis model was used to evaluate anti-tumor efficacy in vivo through emodinanthrone monotherapy and combined with anti-PD-1 treatment. Proteomic and phosphoproteomic analyses, combined with blocking and rescue experiments, were employed to investigate the underlying mechanism. Results The natural product emodinanthrone (EA), a precursor of emodin, was first identified as a potential T-cell immunomodulator. Mechanistic studies revealed that EA exerts immunomodulatory effects on T cells to eliminate multiple tumor cells, mainly by enhancing the JAK3-STAT1/3 signaling axis, leading to increased expression of granzyme B, perforin, and IFN-γ. In vivo, EA treatment significantly reduced the number of lung metastatic nodules by approximately 72.4% and extended overall survival by approximately 42.8%. Furthermore, combination treatment with anti-PD-1 antibody markedly decreased the metastasis rate from 59% to 12.8% and improved the survival rate from 30% to 70%. Conclusion This study first reveals that emodinanthrone possesses T-cell immunomodulatory anti-tumor activity. In vivo findings further support its development as a promising candidate to improve anti-PD-1 efficacy.

ETHNOPHARMACOLOGICAL RELEVANCE:Kadsura longipedunculata is a Yi ethnic herb for hepatic disorders treatment in China. However, its effects on herb- and chemical-induced liver injury remain unknown. AIM OF THE STUDY:To validate traditional ethnomedicinal applications of K. longipedunculata and characterize its bioactive constituents. MATERIALS AND METHODS:The hepatoprotective effect was evaluated using a triptolide-induced mice liver injury model. Bio-guided isolates were screened in an acetaminophen (APAP)-induced HepG2 cell model. Underlying signaling pathways were investigated using transcriptomics, network pharmacology, molecular docking, molecular dynamics simulations, and subsequent Western blotting validation. RESULTS:Abnormal biochemical indices, antioxidant defenses, and liver pathological necrosis in triptolide-induced mice were all adjusted by treatment with K. longipedunculata extract, exhibiting effects superior to those of the positive control, silymarin. The PI3K/AKT/NF-κB signaling pathway was identified as a core therapeutic pathway through transcriptomic profiling, supported by the upregulation of phosphorylated AKT and MTOR concurrent with the downregulation of NF-κB and TNF-α. Four compounds were isolated, among which catechin-5-O-β-D-glucopyranoside (Catechin GP) showed potent anti-apoptotic and antioxidative bioactivities in APAP-induced hepatocytes through the same pathway. The formation of a stable Catechin GP-AKT1 complex was further supported by computational analyses of binding stability and safety. CONCLUSION:The protective effect of K. longipedunculata and its bioactive compound, Catechin GP, against liver injury triggered by both herbal (triptolide) and chemical (acetaminophen) agents is regulated via the PI3K/AKT/NF-κB pathway.

Chemical investigation of Veratrum nigrum L. (Melanthiaceae) resulted in the isolation of three previously undescribed steroidal alkaloids (1-3) and sixteen known analogues (4-19), including a solanidine-type steroidal alkaloid bearing a 3-O-β-D-glucosyl moiety (1) and a 15-hydroxylated jervine-type steroidal alkaloid (2). The absolute configurations of the undescribed compounds were determined by single-crystal X-ray diffraction, DP4+, and ECD calculations. Evaluation of the anticoagulant activity of all isolated compounds revealed that alkaloid 3 prolonged activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) and moderately inhibited rabbit platelet aggregation induced by adenosine diphosphate (ADP) and arachidonic acid (AA).

Phellinus tremulae is a valued mushroom with dual medicinal and edible properties. A chemical investigation on P. tremulae led to the isolation and structural elucidation of eight previously undescribed sesquiterpenoids, named phellitremulins A-H (1, 2, and 5-10), along with six known analogues. Compound 5 features a unique dimeric scaffold linked by two fomannosane units, while 6 should be a key intermediate in the biosynthetic pathway leading to protoilludane sesquiterpenoids. Compounds 1-5 notably inhibited NO production in LPS-stimulated RAW264.7 cells, with IC50 ranging from 1.1 to 6.6 μM. Particularly, compounds 1 and 5 markedly reduced the secretion of the pro-inflammatory cytokines TNF-α and IL-6, and significantly downregulated the protein expression level of iNOS and COX-2 in activated macrophages. Furthermore, 1 and 5 could effectively ameliorate acute inflammation in λ-carrageenan-induced mouse paw edema model. This study reveals that P. tremulae is rich in protoilludane-derived sesquiterpenoids and holds potential for anti-inflammatory applications.

BACKGROUND:Fungal diseases affect over one billion individuals worldwide and represent a significant public health concern. Oral candidiasis (OC) is a common fungal infection primarily caused by Candida albicans. Eriocapitella rivularis is a traditional medicinal herb for the treatment of oral fungal infections. PURPOSE:This study aimed to identify potent antifungal compounds from E. rivularis and elucidate their underlying mechanisms of action. METHODS:Bio-guided isolation combined with network pharmacology was used to identify anti-C. albicans compounds from E. rivularis, followed by metabolomics analysis to explore potential regulatory pathways. Antifungal activity and synergistic efficacy were evaluated using broth microdilution, checkerboard, and time-kill assays. Biofilm inhibition was assessed through quantitative and biochemical analyses. Reverse transcription quantitative polymerase chain reaction was performed to elucidate molecular mechanisms. An in vivo murine OC model was employed to verify efficacy. RESULTS:Prosapogenin CP4 disrupted the fungal cell wall, targeted ergosterol to induce plasma membrane permeabilization, inactivated mitochondrial function, and inhibited hyphal growth, potentially through interference with ABC transporters, modulation of cAMP export, and induction of metabolic disorders. Prosapogenin CP4 and ursolic acid exhibited a synergistic inhibitory effect against fluconazole-resistant C. albicans (FICI = 0.375) with low resistance. This combination regulated genes in the Ras/cAMP/PKA signalling pathway, significantly inhibited biofilm formation and reduced fungal burden, inflammatory responses, and fungal colonisation in murine oral tissues, with efficacy comparable or superior to that of nystatin. CONCLUSION:The finding presents a synergistic phytomedicine strategy using two triterpenoids from E. rivularis as a promising therapeutic approach for OC, particularly biofilm-associated infections.

Two previously undescribed lathyrane-type diterpenoids, daphngenoids A-B (1-2), and one new daphnane-type diterpenoid, daphngenoid C (3), along with eight known compounds (4-11), were isolated from the buds of Daphne genkwa. Notably, daphngenoids A-B (1-2) significantly inhibited doxorubicin-induced inflammation and oxidative stress in the human kidney proximal tubular epithelial cell line (HK-2).

Velbanamines A-L (1-12), 12 novel quebrachamine-like indole alkaloids, were isolated from Catharanthus roseus L. (G.) Don under the guidance of HSQC-based DeepSAT. Their structures and absolute configurations were elucidated unambiguously by comprehensive spectroscopic analyses, single-crystal X-ray diffraction analysis, and electronic circular dichroism (ECD) calculations and computational calculations. Structurally, compound 1 features a unique skeleton with a 6/5/8/5 tetracyclic system. The plausible biosynthetic pathway of 1 was also proposed. In addition, all compounds demonstrated anti-ferroptotic activity in cardiomyocytes. Among them, Compound 8 was the most potent (IC50 = 0.12 ± 0.01 μM), and also suppressed lipid ROS accumulation and regulated ferroptosis-related proteins.

Antibiotic-induced depletion of the gut microbiota facilitated the colonization of vancomycin-resistant Enterococci (VRE) in the gastrointestinal tract, and then increased patients' susceptibility to secondary infections. Ellagic acid, a major constituent of fruits and nuts, showed various bioactivities except for antibacterial. Interestingly, it promoted the recovery of gut microbiota, enhanced microbial diversity and stimulated the proliferation of probiotic gut microbes, and then ameliorated the overgrowth of pathogens in vivo in our experiment. Moreover, ellagic acid activated Gpr41 and Gpr43 mainly by promoting the production of short chain fatty acids (SCFAs) such as acetic acid and propionic acid to inhibit the NF-ĸB signaling pathway. Then the dietary supplement with ellagic acid might treat infected gut to avoid antibiotic-associated intestinal diseases, and the finding also provided a novel strategy for exploring antibacterial agent besides screening in vitro.

Alstoschoquinolines A-D (1-4) representing three unprecedented scaffolds were isolated from the leaves of Alstonia scholaris through direct separation by LC/MS detection. 1 and 2 consisted of a 5/6/5-coupled quinoline architecture containing six consecutive chiral carbons, while 3 and 4 possessed a bridged ring featuring 6/6/6/6 and 6/6/8/6 skeletons, respectively. They might be derived from the corynantheine-type indole alkaloid via sequential oxidation and rearrangement. Compound 3 exhibited a significant inhibitory effect on colon carcinoma cells by disturbing glutathione circulation.

Alstoscholarisine M is a 6,7-seco-vallesamine monoterpenoid indole alkaloid with a rare methylcarbamate fragment, isolated from the leaves of Alstonia scholaris. Extensive spectroscopic data analyses, quantum chemical computations, and single-crystal X-ray diffraction elucidated its structure. The fascinating compound exhibited potential antifungal activity against Trichophyton rubrum, and its activity was roughly comparable to the first-line antifungal drug griseofulvin and significantly more effective than berberine, an important antimicrobial drug originating from plants.