Since its conceptual inception in 1995, Molecular Pharmacognosy has evolved over three decades from a nascent idea into a frontier interdisciplinary discipline characterized by a comprehensive theoretical system, mature methodologies, and extensive applications. This paper systematically reviews the significant milestones of this discipline in theoretical innovation, technical standardization, industrial application, and platform construction. It highlights breakthrough achievements in areas such as technical standards for the molecular identification of crude drugs, elucidation of the formation mechanisms of Dao-di herbs, establishment of conservation models for rare and endangered resources, and biosynthesis of active compounds. The paper further elucidates the evolutionary research paradigm of Molecular Pharmacognosy. Originating from phenomena observed in both the discipline and the industry, this paradigm advances through a progressive cycle of problem derivation—hypothesis presupposition—technical innovation—theoretical enrichment, driven by the dynamic interplay between theory and practice. Furthermore, the paper analyzes current challenges regarding the precision of molecular identification, the complex mechanisms of geo-authenticity, and the sustainable utilization of resources. Molecular Pharmacognosy is poised to integrate deeply with frontier technologies such as artificial intelligence, pan-omics, quantum computing, biomanufacturing, smart breeding, and gene editing. This integration will drive a revolutionary shift in the research paradigm from recognition and revelation to prediction and design. Ultimately, these achievements will mark a new phase defined by precision, systematization, and engineering, injecting fresh momentum into the modernization of TCM.

Focused on the challenges in brain disease treatment and advancements in nose-to-brain delivery research, this article summarized current progress in nasal drug delivery research and proposed several novel evaluation models for nasal drug absorption. By reviewing five types of evaluation models-ex vivo animal tissue models, in situ perfusion models, in vivo pharmacokinetic models, in vitro cell models, and computational fluid dynamics model-it clarified the distinct advantages of nasal drug delivery compared to other administration routes. Furthermore, by outlining emerging nasal drug delivery technologies and irritation evaluation methods, it explored the cutting-edge directions of nose-to-brain delivery. This provides essential support for the selection of nose-to-brain delivery dosage forms and paves new paths for the advancement of nasal drug delivery.

Osteoarthritis (OA) is a common degenerative joint disease characterized by cartilage degeneration, synovial inflammation, osteophyte formation, and joint space narrowing, seriously impairing patients' mobility and quality of life. The pathogenesis of OA is highly complex, involving multiple cell types and signaling pathways. Key pathological processes include abnormal chondrocyte function, sustained activation of inflammatory responses, and imbalances in apoptosis and autophagy. In recent years, microRNAs (miRNAs), small non-coding RNAs, have been recognized for their crucial roles in regulating gene expression and influencing the progression of various diseases. miRNAs exhibit important regulatory functions in OA-related processes such as inflammatory regulation, chondrocyte apoptosis and autophagy, extracellular matrix metabolism, and subchondral bone remodeling. Multiple specific miRNAs have been shown to be closely associated with the onset and progression of OA, demonstrating potential diagnostic and therapeutic value. Traditional Chinese medicine (TCM) is an important treatment approach for OA, offering advantages such as multi-component, multi-target, and multi-pathway regulation. Recent studies have shown that TCM single herbs, compound prescriptions, acupuncture, and moxibustion can upregulate or downregulate the expression of specific miRNAs, thereby intervening in their downstream target genes and related signaling pathways. This leads to anti-inflammatory and analgesic effects, inhibition of chondrocyte apoptosis, promotion of matrix synthesis, and cartilage repair. This review systematically summarizes the mechanisms of miRNAs in the progression of OA, with a focus on the latest research advances in TCM regulation of miRNAs to intervene in OA. The aim is to provide a theoretical basis and new insights for the early molecular diagnosis, targeted therapy, and modern research and clinical translation of TCM in OA.

Colorectal cancer (CRC) is a highly prevalent malignant tumor worldwide, and its occurrence and development are closely related to gut microbiota disturbance. The germ-free (GF) animal model, by providing a unique environment free of indigenous microorganisms, has become a tool for precisely dissecting the causal role and mechanism of the gut microbiota in the occurrence and development of CRC and the therapeutic mechanisms of pharmaceuticals. This review aims to systematically elaborate on the application progress of GF animal models in CRC research, with a focus on the key mechanisms by which specific microorganisms promote the occurrence and development of CRC revealed by this model. Furthermore, this review focuses on the research dynamics of TCM in this field and systematically expounds the application of GF models in the research of TCM against CRC. Through in-depth analysis of the GF animal models of CRC, it is possible to develop precise diagnostic and therapeutic strategies based on TCM and the microbiome in the future, providing new directions and ideas for the prevention and treatment of CRC.

Bacopa monnieri, a plant of Scrophulariaceae, has a long history of medicinal use and is recognized as a traditional medicinal herb in multiple countries. Neurological disorders, due to their extensive damage to neural functions, have become a major public health challenge and garnered worldwide attention. In ancient times, B. monnieri was documented for treating cognitive and memory impairments. Recent studies have elucidated its pharmacological mechanisms and therapeutic targets underpinning the neuroprotective effect. However, the molecular mechanisms of its active ingredients have not been systematically summarized. This study is the first to integrate the target sites of its core ingredients across multiple categories of neurological disorders. It summarizes the pharmacological mechanisms of B. monnieri as well as its active ingredients and derivatives in various neurological diseases, which exert neuroprotective effects through multi-target interactions. This paper provides novel strategies for developing botanical drugs targeting diseases like Alzheimer's and epilepsy and aims to offer evidence-based guidance for the development and application of B. monnieri in functional foods, health supplements, and pharmaceuticals.

Endothelial-to-mesenchymal transition (EndMT) is a process in which endothelial cells lose specific markers and acquire a mesenchymal phenotype under pathological stimulation, contributing to the occurrence and progression of atherosclerosis (AS). Recent studies have shown that TCM, with its unique advantages of holism and treatment based on syndrome differentiation, can inhibit AS by regulating EndMT-related signaling pathways at multiple levels, through multiple pathways and targets. This article systematically reviews the mechanisms of EndMT in AS, as well as the research progress of TCM monomers and compounds in counteracting AS via EndMT intervention, providing new ideas and a scientific basis for the prevention and treatment of AS with TCM.

Epilepsy is a common neurological disorder characterized by refractoriness and recurrence. In China, the prevalence rate is 0.4%-0.7%, with currently over nine million patients, among whom approximately 30% suffer from refractory epilepsy. Although antiepileptic drugs (AEDs) are the mainstay of treatment, they are associated with multi-system adverse reactions and drug resistance issues. In contrast, traditional Chinese medicine (TCM) treatment exhibits significant efficacy with lower toxicity and side effects, and integrated TCM and western medicine treatment can achieve complementary advantages. Studies have shown that integrated therapy demonstrates benefits in various types of epilepsy, including status epilepticus, post-stroke epilepsy, post-traumatic epilepsy, pediatric epilepsy, refractory epilepsy, and epilepsy comorbid with depression and anxiety. It exerts synergistic effects by enhancing AED bioavailability (borneol improves blood-brain barrier permeability), reducing seizure frequency, preventing recurrence, and minimizing western medicine toxicity (Dingxian Decoction decreases digestive/neurological adverse reactions). The underlying mechanisms involve regulating neurotransmitters (modulating glutamate and γ-aminobutyric acid levels) and ion channels (inhibiting sodium/calcium ion channels). In terms of preventive care, TCM plays an important role in preventing epileptic seizures by correcting biased constitutional states. In the future, integrated TCM and western medicine in epilepsy prevention and treatment will move toward individualized treatment, etiological treatment, preventive rehabilitation, and integration of scientific research with clinical practice, forming a three-dimensional development pattern of "technology empowerment, standard guidance, and whole-process management".

Clarifying the two-dimensional spatial distribution and the seasonal yield dynamics of Epimedium pubescens roots in agroforestry systems can provide an ecological theoretical basis and technical support for optimizing the understory cultivation patterns of E. pubescens. E. pubescens plants cultivated in Ginkgo biloba forests (YX), Phellodendron amurense forests (HB), and monocropping fields (CK) were taken as research subjects. Stratified sampling was conducted during the summer and winter harvest seasons to analyze root spatial distribution, morphological characteristics, and biomass allocation. Additionally, high-performance liquid chromatography (HPLC) was employed to determine the total flavonol glycoside content. The results demonstrated that the spatial distribution of E. pubescens roots exhibited significant differences among planting patterns. Specifically, the HB group showed a significantly higher proportion of fine roots in the surface soil layer (0-10 cm) than the YX and CK groups, whereas the YX group displayed more extensive root distribution in deeper soil layers (30-50 cm) than the HB and CK groups. The root morphology of E. pubescens was significantly influenced by planting patterns and seasonal variations. In summer, the root length density of E. pubescens in the YX group decreased by 56%-75% (P<0.05) in the 0-20 cm soil layer compared with that in the CK group, whereas in winter, both agroforestry systems showed significantly higher root length density than the CK group. No significant difference was observed in aboveground or belowground biomass of E. pubescens among different planting patterns. However, the YX group exhibited significantly higher root biomass allocation ratio and significantly lower rhizome biomass allocation ratio than the CK and HB groups. These findings indicate that E. pubescens roots exhibit remarkable phenotypic plasticity, enabling adaptive adjustment strategies to enhance resource acquisition efficiency in agroforestry systems.

Using the whole-genome data of Atractylodes lancea, members of the auxin (Aux)/indole-3-acetic acid (IAA) gene family were identified and characterized, and the effects of 1-naphthaleneacetic acid (NAA) treatment on the expression levels of this gene family were investigated, providing a foundation for further studies on the functions of the A. lancea Aux (AlAux)/IAA gene family. Members of the AlAux/IAA gene family were identified from the A. lancea whole-genome data using TBtools-Ⅱ. Bioinformatics software was employed to analyze the physicochemical properties of the encoded proteins, construct a phylogenetic tree, and predict conserved motifs and gene structures. Tissue-cultured seedlings of A. lancea were used as experimental materials, and key genes of the AlAux/IAA gene family were screened by transcriptome sequencing. The expression of key AlAux/IAA genes was analyzed after one month of NAA treatment. A total of 53 AlAux/IAA genes were identified from the A. lancea genome. Most AlAux/IAA proteins were localized in the nucleus, lacked transmembrane domains, and exhibited negative hydrophobicity. Among them, 44 AlAux/IAA genes were predicted to encode functional Aux/IAA transport proteins. Transcriptome sequencing and RT-qPCR analyses showed that the AlAux/IAA family genes exhibited spatiotemporal expression specificity, and different members displayed varying response times and intensities to NAA treatment. Tissue-specific expression analysis revealed that the AlAux/IAA gene family was generally highly expressed in flowers. This study identified the Aux/IAA gene family members in A. lancea, preliminarily analyzed their sequence characteristics, and examined their expression features in response to Aux. It is speculated that AlAux/IAA13 and AlAux/IAA38 may be involved in rooting and bud growth and development during the subculture process of A. lancea tissue-cultured seedlings. These results provide a theoretical basis for elucidating the molecular mechanisms of AlAux/IAA gene function and Aux-mediated regulation of rhizome growth in A. lancea.

Based on supramolecular chemistry of TCM, this study investigated the interaction between Cinnamomi Cortex-Coptidis Rhizoma herb pair and compared phase states and bioactivity differences under different decoction methods (co-decoction vs individual decoction followed by mechanical mixing). Phase state variations were observed macroscopically and through turbidimetry. The herb interaction was analyzed using isothermal titration calorimetry, UV and IR spectroscopy. Particle size differences were determined by dynamic light scattering and scanning electron microscopy (SEM). Antimicrobial activity was evaluated through broth dilution, plate coating, live/dead staining, and SEM. Component analysis was conducted using UPLC-MS/MS. Results revealed distinct supramolecular structures in Cinnamomi Cortex-Coptidis Rhizoma decoctions, with co-decoction showing more stable macroscopic phase states. The interaction was a spontaneous exothermic reaction driven by both entropy and enthalpy. UV and IR spectra revealed co-decoction demonstrated more sufficient molecular interactions, more stable supramolecular structures, and smaller particle sizes with a more centralized distribution. In addition, co-decoction resulted in higher berberine and coptisine content and stronger inhibitory activity against Staphylococcus aureus compared to mechanical mixtures. These findings indicated that co-decoction produced Cinnamomi Cortex-Coptidis Rhizoma supramolecular systems with more homogeneous phase states, enhanced measurable coptisine and berberine content, and superior antibacterial efficacy.

This study investigated the toxicological mechanisms of different processed products of Strychnos nux-vomica seeds using serum metabolomics analysis. Seventy-two female KM mice were randomly divided into six groups (n = 12 per group): control, raw seeds, vinegar-processed seeds, urine-soaked seeds, sand-roasted seeds, and oil-fried seeds. Each processed preparation group received an aqueous decoction of the original herb at the same concentration for 7 days. Hepatic and renal coefficients were measured in each group, and the levels of hepatic and renal injury markers, including aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN), and creatinine (CRE) were determined using a fully automated biochemical analyzer. Hematoxylin-eosin (HE) staining was used for histopathological examination of the liver and kidney. Liquid chromatography-mass spectrometry (LC-MS/MS) combined with multivariate statistical analysis was employed to screen for differential serum metabolites across groups. Metabolic pathway enrichment analysis of the selected differential markers was performed using the Human Metabolome Database (HMDB) and MetaboAnalyst 6.0. All processed S. nux-vomica seed products increased hepatic and renal coefficients and elevated hepatic and renal injury markers (AST, ALT, BUN, CRE), although these effects were reduced to varying degrees compared to the raw product. Overall, vinegar processing exhibited the most significant detoxification effect, followed by oil-fried>sand-roasted≈urine-soaked > raw seeds. Metabolomic analysis identified 14 differential metabolites across the groups, primarily including sphingolipids, glycerophospholipids, and vitamin-related compounds. Five significantly perturbed metabolic pathways were identified: sphingolipid metabolism, glycerophospholipid metabolism, α-linolenic acid metabolism, lipoic acid metabolism, and linoleic acid metabolism, with sphingolipid and glycerophospholipid metabolism common to all groups. In conclusion, processing of S. nux-vomica seeds reduced hepatorenal toxicity to varying degrees, and its toxic mechanisms may primarily involve disturbances in sphingolipid and glycerophospholipid metabolism.

The stem bark of Aquilaria yunnanensis (Thymelaeaceae) is traditionally used to treat cough and asthma in folk medicine. However, its active components have not been reported. In the present study, the chemical constituents of A. yunnanensis stem bark were isolated and identified, and their inhibitory activities against nitric oxide (NO) production in RAW264.7 cells induced by lipopolysaccharide (LPS) were evaluated to clarify the anti-inflammatory components in the stem bark of A. yunnanensis. A total of 22 compounds, including a new norlignan [(±)-1], were isolated from the ethanolic extract of A. yunnanensis stem bark by column chromatography, high-performance liquid chromatography (HPLC), recrystallization, etc. Through chiral separation of (±)-1, (-)-1 and (+)-1 were obtained. Based on electronic circular dichroism (ECD) calculations, the absolute configurations of (-)-1 and (+)-1 were determined to be (-)-(7R, 8S, 8′S)-4-O- (1, 3-dihydroxypropan-2-yl) zhebeiresinol and (+)-(7S, 8R, 8′R)-4-O- (1, 3-dihydroxypropan-2-yl) zhebeiresinol. The known compounds included (±)-zhebeiresinol (2), (+)-syringaresinol (3), (+)-episyringaresinol (4), orcinol (5), orsellinic acid (6), methyl orsellinate (7), 3, 4-dihydroxybenzoic acid (8), lecanoric acid (9), barbatic acid (10), 7β-hydroxy-β-sitosterol (11), β-sitosterol (12), stigmasterol (13), ergosterol peroxide (14), genkwanin (15), pilloin (16), 5-hydroxy-3′, 4′, 7-trimethoxyflavone (17), (-)-epicatechin (18), (-)-epicatechin gallate (19), tectorigenin 2-O-α-L-rhamnopyranoside (20), 2, 3-dihydroxypropyl docosanoate (21), and 1-O-linolenoylglycerol (22). According to the bioassay testing results, compounds 4 and 22 which inhibited NO production with IC₅₀ values of (32.82±1.04) and (18.50±0.95) μmol·L^-1, respectively, exhibited anti-inflammatory activities.

In this study, the total alkaloids of Corydalis delicatula were isolated and purified by a combination of silica gel, aluminum oxide basic column chromatography and other chromatographic techniques. The obtained compounds were structurally identified with the help of modern spectral resolution techniques such as nuclear magnetic resonance (1D/2D NMR), high-resolution mass spectrometry (HR-ESI-MS), and single crystal X-ray diffraction. Nine alkaloids were obtained from the total alkaloids of C. delicatula, encompassing two new compounds, corlicatine (1) and N-methyl-13-oxoprotopine (2), and seven known ones: (+)-ochotensine (3), rupestrine A (4), (+)-ochotensimine (5), oxohydrastinine (6), α-magnoflorine (7), 6, 7-methylenedioxy-N-methylisoquinoline (8), and N-methyl-6, 7-dimethoxyisoquinoline (9). The inhibitory activities of the compounds against acetylcholinesterase were evaluated, with compound 4 exhibiting moderate inhibition. Molecular docking was further performed to elucidate the binding mode of this compound. The results provide a basis for elucidating the medicinal substance basis of C. delicatula.

The chemical constituents from Sanguisorba officinalis were isolated and purified through macroporous resin, silica gel, polyamide, ODS, Sephadex LH-20 column chromatography, and preparative HPLC. Their structures were determined by extensive analysis of data from MS, NMR, OR, and calculated NMR and ECD. Twenty-six compounds (1-26) were isolated from the 95% ethanol extract of S. officinalis and identified as (1S, 4S, 5R, 7S, 10R, 11R)-1α, 11, 12-trihydroxy-eudesmane (1), vomifolilol (2), (3S, 5R, 6R, 7E)-3, 5, 6-trihydroxy-7-megastigmen-9-one (3), threo-1, 2-bis- (4-hydroxy-3-methoxyphenyl)-propane-1, 3-diol (4), erythro-1, 2-bis- (4-hydroxy-3-methoxyphenyl)-propane-1, 3-diol (5), phenylformic acid (6), vanillic acid (7), veratric acid (8), 3, 4-dimethoxy-5-hydroxybenzoic acid (9), 4- (1-hydroxy-1-methylethyl)-benzoic acid (10), p-hydrocoumaric acid (11), 4-hydroxyphenethyl alcohol (12), ω-hydroxypropioguaiacone (13), cytosporone V (14), indazole (15), 1H-indole-3-carboxylic acid (16), 7, 8-dimethyl-iso-alloxazine (17), (-)-jasmonic acid (18), 12-hydroxyjasmonic acid (19), (-)-methyl 12-hydroxyjasmonate (20), 5-hydroxyhexan-4-olide (21), (8R^*, 9R^*, 10S^*, 6Z)-trihydroxyoctadec-6-enoic acid (22), (9S, 10R, 11E, 13R)-9, 10, 13-trihydroxyoctadec-11-enoic acid (23), methyl (9S, 10R, 11E, 13R)-9, 10, 13-trihydroxyoctadec-11-enoate (24), pinellic acid (25), and (9E)-8, 11, 12-trihydroxyoctadecenoic acid methyl ester (26). Compound 1 was a new eudesmane-type sesquiterpenoid and named sanguisorbaol M. Compounds 3-5, 8, 10, 14-17, 19-23, 25, and 26 were isolated from Rosaceae for the first time. Compounds 2, 11, 12, 18, and 24 were firstly isolated from the genus Sanguisorba and compound 6 was isolated from S. officinalis for the first time. The inhibitory effects of the isolated compounds on melanin synthesis were evaluated via the 3-isobutyl-1-methylxanthine (IBMX)-induced B16F10 melanoma cell model. The results showed that compounds 1-3 at a concentration of 50 μmol·L^-1 had a significant anti-melanogenic activity.

This study established a hyperlipidemia model by feeding Sprague-Dawley rats a high-fat diet for 8 weeks. The rats were randomly assigned to the following groups: model group, atorvastatin calcium group (4.8 mg·kg^-1), low-, medium-, and high-dose Tanyu Tongzhi Optimization Decoction (TYTZD) groups (3.6, 7.2, and 14.4 g·kg^-1), and a normal diet control group. After 4 weeks of continuous administration, hematoxylin-eosin (HE) and oil red O staining were used to observe liver pathological changes and lipid infiltration. Automatic biochemical analyzer were performed to assess blood lipid profiles, coagulation function, and liver function. Transcriptomic and proteomic analyses were employed to identify differentially expressed genes (DEGs) and proteins (DEPs), followed by enrichment analysis. The MCODE algorithm was applied to classify DEGs and DEPs into modules, and network separation index (S_AB) was calculated to assess module separation, enabling construction of a gene-protein co-expression network for core target screening. The diagnostic accuracy of core targets was evaluated by area under the receiver operating characteristic (ROC) curve (AUC), and ELISA was used to measure core target expression. Western blot detected the expression of core pathway-related proteins in liver tissue. Results demonstrated that TYTZD significantly improved dyslipidemia, coagulation dysfunction, liver injury, hepatic pathology, and lipid infiltration in hyperlipidemic rats. Transcriptomic analysis identified 571 DEGs significantly reversed by TYTZD, mainly enriched in inflammatory signaling pathways such as Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB). Proteomic analysis identified 102 reversed DEPs, mainly involved in cholesterol metabolism pathways. Integrated analysis identified core targets including TLR4, tumor necrosis factor-α (TNF-α), integrin subunit alpha M (ITGAM), Toll-like receptor 2 (TLR2), matrix metalloproteinase 9 (MMP9), interleukin-1β (IL-1β), apolipoprotein E (APOE), and apolipoprotein C2 (APOC2), all with AUC values greater than 0.70. ELISA showed that TYTZD intervention significantly downregulated MMP9, TNF-α, IL-1β, TLR2, ITGAM, and TLR4, and upregulated APOC2 and APOE. Western blot indicated that TYTZD reduced TLR4, p-NF-κB, and IL-1β protein expression in liver tissue. In conclusion, TYTZD may exert anti-hyperlipidemic effects through regulation of core targets such as ITGAM, TLR4, and APOC2, and by modulating the TLR4/NF-κB signaling pathway to intervene in inflammatory responses and cholesterol metabolism, thereby achieving multi-target, multi-pathway therapeutic effects against hyperlipidemia.

This study investigated the asthma-alleviating effects of Poria cocos from the perspectives of immuno-inflammation and gut microbiota (GM), providing a theoretical basis for its rational clinical use. Male Balb/c mice were randomly divided into normal, model, positive control (montelukast sodium, 1.5 mg·kg^-1), low-dose P. cocos (0.057 g·kg^-1), medium-dose P. cocos (0.114 g·kg^-1), and high-dose P. cocos (0.227 g·kg^-1) group. Except for the normal group, the other groups were sensitized with ovalbumin (OVA) by intraperitoneal injection combined with nebulization to establish an asthma model, and treated with the corresponding drugs. After modeling, cough and wheezing indices, lung function, and asthma characteristic indicators including immunoglobulin E (IgE), interferon-γ (IFN-γ), interleukin 4 (IL4), and interleukin 10 (IL10) levels were measured. Lung tissue pathology was observed by hematoxylin-eosin (HE) and Masson staining. Quantitative real-time PCR (qRT-PCR) was used to detect airway inflammation-and remodeling-related mRNA levels in lung tissue. Levels of 12 inflammatory factors in serum and bronchoalveolar lavage fluid were detected by multiplex assay. Furthermore, the asthma-alleviating mechanism of P. cocos was analyzed by network pharmacology and GM 16S rRNA sequencing. Results showed that P. cocos reduced cough frequency in asthmatic mice, improved lung function, alleviated pathological damage in lung tissue, decreased serum IgE and IL 4 levels, increased IL 10 and IFN-γ levels, and multi-target regulated related inflammatory factors in serum and alveolar lavage fluid, maintaining immune homeostasis. Network pharmacology predicted that P. cocos exerts its asthma-alleviating effects via inflammation regulation pathways. GM sequencing showed that P. cocos effectively alleviated dysbiosis induced by OVA sensitization, significantly downregulated Muribaculum intestinale and Alistipes inops, and correlation analysis revealed both were positively correlated with pro-inflammatory factors. Therefore, it is speculated that P. cocos restores intestinal microecological homeostasis by reducing levels of M. intestinale and A. inops, decreases pro-inflammatory factor production, blocks the inflammatory cascade, and thereby exerts its asthma-alleviating effects.

This study investigates the inducing effects of jatrorrhizine (JAT), a bioactive compound extracted from Coptidis Rhizoma, on the p53-mediated ferroptosis in human colorectal cancer (CRC) cell lines HT29 and HCT116, specifically through the modulation of spermidine/spermine N1-acetyltransferase 1 (SAT1). Cell viability was assessed via the cell counting kit-8 (CCK-8) assay, while the colony formation assay was employed to determine the proliferative capacity. Cell cycle distribution was analyzed by flow cytometry, and mitochondrial membrane potential was evaluated via JC-1 dye. High-throughput RNA sequencing (RNA-seq) was performed to analyze transcriptomic changes in JAT-treated HT29 and HCT116 cells. Lentiviral transduction was utilized to generate SAT1-overexpressing HCT116 and HT29 cell lines, with transfection efficiency confirmed by qPCR. The levels of intracellular Fe²⁺, reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) were quantified via specific commercial assay kits. The protein and mRNA levels of key ferroptosis-related markers: solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), tumor protein p53, and SAT1 were determined by Western blot and qPCR, respectively. The results demonstrated that JAT treatment significantly inhibited both cell viability and proliferation, induced cell cycle arrest, and reduced mitochondrial membrane potential in HCT116 and HT29 cells. RNA-seq results revealed that the KEGG pathways most significantly enriched in JAT-treated cells were protein digestion and absorption, arginine and proline metabolism, glycine, serine and threonine metabolism, and ferroptosis. Notably, the mRNA level of SAT1 in the arginine metabolism pathway was significantly upregulated following JAT treatment. Furthermore, JAT treatment increased the levels of Fe²⁺, ROS, and MDA, while markedly decreasing GSH content in a dose-dependent manner. In addition, the treatment downregulated the protein levels of the ferroptosis inhibitors SLC7A11 and GPX4, and upregulated the expression of the pro-ferroptotic proteins ACSL4, SAT1, and p53. SAT1 overexpression inhibited cell proliferation and activated the p53-mediated ferroptosis signaling pathway. The combination of SAT1 overexpression and JAT treatment (oveSAT1+JAT group) exhibited a synergistic effect, showing superior efficacy over SAT1 overexpression alone (oveSAT1 group). In conclusion, JAT activates the p53-dependent ferroptosis signaling pathway by upregulating the key metabolic enzyme SAT1, which is involved in proline metabolism, thereby inhibiting colorectal cancer cell proliferation.

This study investigated the effects and molecular mechanisms of sarsasapogenin (SSG) on colorectal cancer (CRC). After HRT-18 and HCT116 cells were treated with varying concentrations of SSG, the effect of SSG on CRC cell proliferation, migration, and glycolysis was assessed by cell counting kit-8 (CCK-8), real-time cellular analysis (RTCA), colony formation assay, high-content imaging analysis, and Seahorse glycolysis stress test. Changes in intracellular lactate release and adenosine triphosphate (ATP) levels under SSG treatment were measured by lactate and ATP detection kits, respectively. The effects of SSG on the transcription level and protein expression of the key glycolytic gene hypoxia-inducible factor 1α (HIF1α) were evaluated via quantitative polymerase chain reaction (qPCR) and Western blot. Direct binding between SSG and HIF1α was identified by drug affinity responsive target stability (DARTS). The UbiBrowser database, microscale thermophoresis (MST), and molecular docking simulations were employed to screen and validate the E3 ubiquitin ligase mediating HIF1α degradation. The results showed that SSG significantly inhibited CRC cell proliferation and colony formation in a dose-dependent manner. The 24-hour half-maximal inhibitory concentrations (IC₅₀) of SSG for HRT-18 and HCT116 cells were 6.275 μmol·L^-1 and 7.029 μmol·L^-1, respectively. After 24 hours of treatment with SSG at concentrations of 2.0, 4.0, and 6.0 μmol·L^-1, the HRT-18 and HCT116 clone formation rate was significantly reduced. High-content imaging results showed that SSG significantly inhibited the migration ability of CRC cells. Seahorse glycolysis stress test, along with lactate and ATP measurements, demonstrated that SSG suppressed basal glycolysis, glycolytic capacity, glycolytic reserve, lactate release, and ATP content in CRC cells. The qPCR and Western blot results showed that SSG had no significant effect on HIF1α mRNA expression, and the HIF1α protein expression was downregulated. DARTS confirmed the direct binding of SSG to HIF1α. UbiBrowser analysis identified five E3 ubiquitin ligases potentially targeting HIF1α. Molecular docking and MST results indicated that the F-box/WD repeat-containing protein 7 (FBXW7) exhibited strong binding affinity to HIF1α in the presence of SSG. In summary, SSG exerts its anti-cancer effects in CRC by targeting HIF1α to promote its FBXW7-mediated proteasomal degradation and consequently inhibiting glycolytic reprogramming.

This study explored the mechanism by which Suanzaoren Decoction (SZRD) ameliorates hippocampal neuronal damage in APP/PS1 mice by inhibiting Bax/Bcl-2/caspase-3-mediated apoptosis through the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Differentially expressed genes (DEGs) in Alzheimer's disease (AD) were screened using the GEO database, and KEGG and GO enrichment analyses were performed using R software. Thirty 6-month-old APP/PS1 transgenic mice were randomly assigned to three groups: APP/PS1 group, low-dose SZRD group (12.96 g·kg^-1), and high-dose SZRD group (25.92 g·kg^-1), while ten age-matched male C57BL/6J mice served as the blank control group. The treatment groups received SZRD by gavage for 28 consecutive days. Following treatment, spatial learning and memory were assessed using the Morris water maze test. Neuronal numbers were evaluated by Nissl staining. Immunohistochemistry (IHC) was used to detect neuron-specific nuclear protein (NeuN) and assess amyloid-β (Aβ) deposition in hippocampal neurons. Western blot was performed to measure the expression levels of PI3K/Akt pathway proteins and apoptosis-related markers, including Bcl-2, Bax, caspase-3, and cleaved-caspase-3 in hippocampal tissues. Bioinformatics analysis identified 686 DEGs, with GO enrichment linked to synaptic function and neurotransmitter transport, and KEGG pathways including neuroactive ligand-receptor interaction and PI3K/Akt signaling. In vivo experiments showed that, compared with APP/PS1 mice, both low-and high-dose SZRD groups exhibited improved learning and memory, increased neuronal numbers, and decreased Aβ deposition. Furthermore, hippocampal levels of p-PI3K/PI3K, p-Akt/Akt, and Bcl-2 were upregulated, while Bax and cleaved-caspase-3/caspase-3 levels were downregulated. In conclusion, SZRD may improve hippocampal neuronal damage and cognitive function in APP/PS1 mice by regulating Bax/Bcl-2/caspase-3-mediated apoptosis through the PI3K/Akt signaling pathway.

This article aims to investigate the inhibitory effect and molecular mechanism of sculponeatin A (STA) on triple negative breast cancer (TNBC). MDA-MB-436 and MDA-MB-468 were selected as cell models. The MTT assay, real-time cell analysis (RTCA), and colony formation assay were used to evaluate the effects of different concentrations of STA on the proliferation of TNBC cells. JC-1 staining and Annexin V-FITC/PI double staining combined with flow cytometry were used to measure the effect of STA on the apoptosis of TNBC cells. Western blot was employed to determine the expression changes of apoptosis-related proteins [cysteinyl aspartate-specific proteinase (caspase)-9, caspase-3, and poly-ADP-ribose polymerase (PARP)] and cancerous inhibitor of protein phosphatase 2A (CIP2A)/protein kinase B (AKT) signaling pathway proteins [CIP2A, AKT, phosphorylated (p)-AKT] in TNBC cells after STA treatment. To clarify the function of CIP2A in the action of STA, a CIP2A overexpression or CIP2A knockdown plasmid was transfected into cells, which were then treated with STA. Cell proliferation, apoptosis, and protein expression changes were evaluated by CCK-8, flow cytometry, and Western blot. Further, RT-qPCR and Western blot both showed that STA significantly downregulated the mRNA and protein levels of CIP2A and the protein level of c-Myc. The overexpression of c-Myc antagonized the downregulating effects of STA on the protein and mRNA levels of CIP2A, while the knockdown of c-Myc enhanced this effect. The drug affinity-responsive target stability (DARTS) assay and microscale thermophoresis (MST) assay confirmed the existence of direct binding between STA and c-Myc protein. The results indicated that STA significantly inhibited the proliferation and colony formation and induced the apoptosis of TNBC cells, manifested by an increased apoptosis rate, downregulated precursor protein expression of caspase-9 and caspase-3, and increased PARP cleavage. STA treatment reduced the p-AKT level but did not affect total AKT. Functionally, the knockdown of CIP2A enhanced the STA effects of inhibiting proliferation and inducing apoptosis, while the overexpression of CIP2A produced antagonistic effects. From a mechanism perspective, STA directly targets and binds to c-Myc to downregulate its expression, thereby inhibiting the transcription and translation of CIP2A and ultimately blocking the c-Myc/CIP2A signaling pathway. In conclusion, STA inhibits the malignant progression of TNBC by targeting the c-Myc/CIP2A signaling axis.

This study aims to investigate whether the mitophagy receptor FUN14 domain-containing 1 (FUNDC1) serves as a molecular link between mitophagy and neuronal ferroptosis, and to determine whether salidroside (Sal) can inhibit neuronal ferroptosis after oxygen-glucose deprivation/reoxygenation (OGD/R) by inducing FUNDC1 expression and regulating mitophagy, thereby exerting neuroprotective effects. An in vitro model of neuronal ischemia-reperfusion injury was established with HT22 cells subjected to OGD/R. The experiment consisted of three parts: ① control, OGD/R, ferrostatin-1 (Fer-1), FUNDC1 overexpression (OV-FUNDC1), and OV-FUNDC1+Fer-1 groups; ② control, OGD/R, 3-methyladenine (3MA, an autophagy inhibitor), Sal, and Sal+3MA groups; ③control, OGD/R, Sal, FUNDC1 silencing (Si-FUNDC1), and Sal+Si-FUNDC1 groups. In the first part of the experiment, the survival rate of cells in each group was detected by the CCK-8 assay, and the protein levels of p62, microtubule-associated protein light chain 3 (LC3), acyl-CoA synthetase long-chain family 4 (ACSL4), and glutathione peroxidase 4 (GPX4) were measured by Western blot. The results showed that compared with the control group, the OGD/R group had down-regulated protein levels of p62 and GPX4 (P<0.05, P<0.01) and up-regulated protein level of ACSL4 (P<0.01); compared with the OGD/R group, the OV-FUNDC1 group showed increased protein levels of LC3 and GPX4 (P<0.05, P<0.01) and decreased protein levels of p62 and ACSL4 (P<0.05, P<0.01). In the second part of the experiment, after Sal intervention, the protein levels of FUNDC1, p62, LC3, ACSL4, and GPX4 were determined by Western blot, and changes in mitochondrial membrane potential were measured via JC-1. The results showed that compared with the OGD/R group, the Sal group had up-regulated protein levels of FUNDC1, LC3, and GPX4 (P<0.01), down-regulated protein levels of p62 and ACSL4 (P<0.01), and increased mitochondrial membrane potential (P<0.01); the 3MA group showed decreased mitochondrial membrane potential (P<0.01). In the third part of the experiment, after silencing of FUNDC1, the intracellular Fe²⁺ content was measured via a ferrous ion assay kit; reactive oxygen species (ROS) levels were measured by flow cytometry; the mitochondrial function was assessed via MitoTracker Red; adenosine triphosphate (ATP) and glutathione (GSH) levels were detected using assay kits; the expression of ACSL4 and GPX4 was detected by immunofluorescence; the protein levels of p62, LC3, ACSL4, and GPX4 were measured by Western blot. The results showed that compared with the Si-FUNDC1 group, the Sal group had decreased Fe²⁺ concentration (P<0.01), reduced ROS level (P<0.01), increased MitoTracker Red fluorescence intensity (P<0.01) and GSH content (P<0.01), weakened ACSL4 fluorescence intensity (P<0.01), enhanced GPX4 fluorescence intensity (P<0.01), up-regulated protein levels of LC3 and GPX4 (P<0.01), and down-regulated protein levels of p62 and ACSL4 (P<0.01). This study reveals that FUNDC1 may be a key protein linking mitochondrial autophagy and ferroptosis in neurons after ischemia-reperfusion injury, and Sal intervention can inhibit neuronal ferroptosis after ischemia-reperfusion by promoting the non-ubiquitinated FUNDC1-dependent mitophagy pathway.

This study observed the effect of Chaihu Jia Longgu Muli Decoction on myocardial fibrosis in the rat model of myocardial infarction (MI) and explored the effect of this decoction on ventricular remodeling induced by myocardial fibrosis after MI based on transforming growth factor-β1 (TGF-β1)/Smad proteins (Smads) pathway, aiming to reveal the therapeutic mechanism of this decoction. MI model rats were established by coronary artery ligation and assigned into sham, model, and low-dose, medium-dose, and high-dose (2.09, 4.19, and 8.37 g·kg^-1, respectively) Chaihu Jia Longgu Muli Decoction groups, with 10 rats in each group. Chaihu Jia Longgu Muli Decoction groups were administrated with the decoction at different doses by gavage, and the sham and model groups received an equal volume of distilled water once daily for 4 weeks. The cardiac function of rats after MI was assessed via echocardiography. Myocardial fiber deposition and collagen volume fraction (CVF) were observed by Sirius red staining and Masson staining. Ultrastructure changes of myocardial cells were observed by transmission electron microscopy. Creatine kinase isoenzymes (CK-MB), cardiac troponin T (cTnT), and soluble growth stimulation expressed gene 2 (sST2) levels were measured by ELISA. Western blot was employed to assess the protein levels of collagen Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) associated with myocardial fibrosis, and TGF-β1, Smad3, and Smad7 associated with the TGF-β1/Smads pathway. Compared with the sham group, the model group had significantly decreased body weight, compromised cardiac function, abnormal ventricular structure, collagen fiber deposition, increased fibrosis area and CVF, seriously damaged ultrastructure of myocardium, a part of mitochondria being swollen or even ruptured, disarrangement and lysis of myofibril, and broken myotome and myofilament with emergence of space. Furthermore, the model group showed risen levels of cTnT, sST2 and CK-MB, obvious myocardial fibrosis, up-regulated protein levels of pro-fibrotic cytokines TGF-β1 and Smad3, down-regulated protein level of the inhibitory cytokine Smad7, and increased expression of myocardial fibrosis-related proteins Col Ⅰ and α-SMA. Compared with the model group, Chaihu Jia Longgu Muli Decoction groups showed increased body weight, improved cardiac function and ventricular structure, reduced collagen fiber deposition, fibrosis area, and CVF, intact ultrastructure of myocardium, normal mitochondrial structure, and well-arranged myofibril and myofilament. In addition, Chaihu Jia Longgu Muli Decoction groups presented declined cTnT, sST2, and CK-MB levels, down-regulated protein levels of TGF-β1 and Smad3, up-regulated protein level of Smad7, and reduced expression of Col Ⅰ and α-SMA. Chaihu Jia Longgu Muli Decoction may reduce excessive deposition of extracellular matrix by regulating the expression of proteins in the TGF-β1/Smads pathway, thereby alleviating myocardial fibrosis and ameliorating ventricular remodeling.

This study explored the mechanism by which the classical prescription Baihe Dihuang Decoction alleviates neuronal injury in anxious depression, based on the Src homology 2 domain-containing protein tyrosine phosphatase (SHP2) signaling pathway. Sprague-Dawley rats were randomly divided into a normal group, model group, venlafaxine group (13.5 mg·kg^-1), and Baihe Dihuang Decoction high-, medium-, and low-dose groups (16, 8, and 4 g·kg^-1, respectively). An anxious depression model was established using 28 days of chronic restraint stress (6 h/day) combined with corticosterone administration (30 mg·kg^-1, ih). From day 8 of modeling, intragastric administration of the corresponding treatments was given for 21 consecutive days. Anxiety-and depression-like behaviors were evaluated. Golgi staining was used to observe dendritic spine morphology of hippocampal neurons, and Western blot was performed to detect hippocampal phosphorylated (p)-SHP2, synapsin 1 (SYN1), and postsynaptic density protein-95 (PSD-95). In cell experiments, primary hippocampal neurons were transfected with lentiviruses to specifically overexpress or knock down SHP2. Transfection efficiency was assessed by RT-PCR and Western blot. A corticosterone-induced neuronal injury model was established, followed by intervention with Baihe Dihuang Decoction-containing serum. Cell viability was measured using the MTT assay, apoptotic morphology was observed by Hoechst 33342 staining, apoptosis rates were determined via flow cytometry, and immunofluorescence was used to detect SHP2, p-SHP2, SYN1, and PSD-95 expression. The results showed that, compared with the normal group, model rats exhibited hippocampal dendritic spines with atrophy, loss, irregular morphology, and shortening, accompanied by significantly reduced SYN1 and PSD-95 expression and markedly increased p-SHP2/SHP2 levels. After Baihe Dihuang Decoction intervention, hippocampal neuronal injury was significantly alleviated, and p-SHP2/SHP2 levels were suppressed. In vitro, neuronal SHP2 overexpression (SHP2 OE) or corticosterone modeling led to a marked decrease in cell viability, increase in apoptosis rate, and downregulation of SYN1 and PSD-95. Following treatment with Baihe Dihuang Decoction-containing serum, SHP2 overexpression was inhibited, cell viability increased, apoptosis decreased, and SYN1 and PSD-95 expression significantly increased. In addition, SHP2 knockdown (SHP2 KD) via lentiviral transfection also mimicked the protective effects of Baihe Dihuang Decoction. In conclusion, Baihe Dihuang Decoction improves hippocampal neuronal injury in anxious depression by regulating the SHP2 signaling pathway.

This study investigates the mechanisms and target sites of Renshen (Ginseng Radix et Rhizoma)-Danshen (Salviae Miltiorrhizae Radix et Rhizoma) (RSDS) in regulating myocardial ischemia-reperfusion injury (MIRI) via the silent information regulator 3 (SIRT3)/peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α)/voltage-dependent anion-selective channel 1 (VDAC1) signaling pathway. A rat MIRI model was established and randomly divided into control, model, fosinopril, and low-, medium-, and high-dose RSDS groups. Cardiac function, cell apoptosis, and myocardial fibrosis were assessed using cardiac ultrasound, TUNEL staining, hematoxylin-eosin (HE) staining, and Masson staining. Serum factors and the expression of key proteins and genes, such as SIRT3, PGC-1α, and VDAC1 in myocardial tissue, were analyzed using ELISA, Western blot, and RT-qPCR to investigate the underlying mechanisms through the SIRT3/PGC-1α/VDAC1 signaling pathway. The experimental results showed that, compared with the model group, RSDS significantly improved cardiac function, with the high-dose group exhibiting the most pronounced cardioprotective effect, including increased left ventricular ejection fraction (LVEF), decreased left ventricular end-systolic diameter (LVESD), and increased left ventricular fractional shortening (LVFS). TUNEL staining demonstrated that RSDS reduced myocardial cell apoptosis, and Masson staining indicated that RSDS significantly alleviated myocardial fibrosis. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), and creatine kinase-MB (CK-MB) were significantly decreased, while superoxide dismutase (SOD) levels were significantly increased, suggesting strong anti-inflammatory and antioxidant effects. Western blot and RT-qPCR analyses demonstrated that RSDS improved mitochondrial function, inhibited oxidative stress, and protected myocardial cells by regulating the SIRT3/PGC-1α/VDAC1 signaling pathway. In conclusion, RSDS significantly ameliorates cardiac dysfunction induced by MIRI through regulation of the SIRT3/PGC-1α/VDAC1 signaling pathway, reducing myocardial apoptosis, fibrosis, and inflammation.

This study aims to investigate the mechanism by which modified Xiangsha Liujunzi Decoction (M-XS) ameliorates hepatic inflammation in hyperlipidemic mice by regulating the intestinal high-density lipoprotein 3 (HDL3) level. Intestine-specific ATP-binding cassette transporter A1 (ABCA1) knockout mice (Abcal-Villin-cre-Tg) were randomized into 3 groups (n = 8): intestine-specific knockout group (Abcal^△Vill), intestine-specific knockout mice fed a high-fat diet group (Abcal^△Vill + HFD), and intestine-specific knockout with Chinese herbal medicine intervention group (Abcal^△Vill + HFD + M-XS). Littermate control mice were randomly allocated into 3 groups (n = 8): blank group (Abcal^fl/fl), high-fat diet group (Abcal^fl/fl+HFD), and Chinese herbal medicine intervention group (Abcal^fl/fl +HFD+M-XS). Except the Abcal^fl/fl and Abcal^△Vill groups, the other groups were fed a HFD for 10 weeks. Drug administration lasted for 6 weeks at a dose of 23.66 g·kg^-1·d^-1. Serum lipid profiles (four items), hepatic histopathology (HE and oil red O staining), and ileal histopathology (HE staining) were assessed. Ileal HDL3 levels were measured by ELISA, hepatic triacylglycerol (TG) levels by the GPO-PAP method, and co-localization of CD86 and Toll-like receptor 4 (TLR4) in the liver tissue by immunofluorescence. The mRNA levels of hepatic lipopolysaccharide-binding protein (LBP), nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-κB p65), inhibitor of nuclear factor kappa-B alpha (IκBα), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were determined by real-time PCR. The protein levels of ABCA1 and apolipoprotein A-I (apoA-I) in the ileum, as well as LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β in the liver, were assessed by Western blot. Compared with the Abcal^fl/fl group, the Abcal^fl/fl +HFD group showed increased blood lipid levels, numerous vacuoles and lipid droplets in the liver, shortened intestinal villi, elevated hepatic TG level, reduced levels of HDL3, ABCA1, and apoA-I in the ileum, co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β as well as protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β in the liver (P<0.05). Compared with the Abcal^fl/fl +HFD group, the Abcal^fl/fl +HFD+M-XS group showed decreased blood lipid levels, reduced vacuoles and lipid droplets in the liver, increased intestinal villus length, decreased hepatic TG level, elevated levels of HDL3, ABCA1, and apoA-I in the ileum, weakened co-localization of CD86 and TLR4 in the liver, and down-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β (P<0.05). Compared with the Abcal^fl/fl+HFD group, the Abcal^△Vill+HFD group showed raised blood lipid levels, increased vacuoles and lipid droplets in the liver, elevated hepatic TG level, enhanced co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, TNF-α, and IL-1β (P<0.05). Compared with the Abcal^fl/fl+HFD+M-XS group, the Abcal^△Vill+HFD+M-XS group showed raised blood lipid levels, increased vacuoles and lipid droplets in the liver, elevated hepatic TG level, enhanced co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β (P<0.05). In conclusion, modified Xiangsha Liujunzi Decoction ameliorates hepatic inflammation and corrects dyslipidemia by modulating intestinal HDL3 levels.

Using bioinformatics analysis, network pharmacology, and cellular experiments, this study investigated the effects and potential mechanisms of Panax notoginseng saponins (PNS) on osteogenic induction of immortalized human aortic valve interstitial cells (hVICs). A calcific aortic valve disease (CAVD) cell model was established by treating hVICs with osteogenic induction medium (OM). Experimental groups included a control group, a model group, and PNS treatment groups (0.075, 0.05, and 0.025 mg·mL^-1). Cell viability was assessed by the CCK-8 assay. Calcification was evaluated by alkaline phosphatase (ALP) and alizarin red S staining. Western blot was performed to measure the expression of calcification-related proteins runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), and ALP. Transcriptomic datasets related to CAVD were obtained from the Gene Expression Omnibus (GEO) database, and CAVD disease targets were retrieved from GeneCards. Active components of PNS and their potential targets were collected from HERB, Batman-TCM 2.0, and ETCM databases. The intersecting genes were subjected to protein-protein interaction (PPI) network construction, Gene Ontology (GO) enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, identifying the spleen tyrosine kinase (SYK)/nuclear factor kappa-B (NF-κB) signaling pathway as a core pathway. Western blot was then used to assess the effects of PNS at the optimal concentration on SYK/NF-κB pathway-related proteins. NF-κB p65 nuclear translocation was examined by immunofluorescence co-localization, and ELISA was applied to measure tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) levels. Results showed that PNS reduced alizarin red S and ALP staining in calcification-induced hVICs and downregulated calcification-related proteins BMP2, ALP, and RUNX2 (P<0.05, P<0.01), with the most significant effect observed at 0.075 mg·mL^-1. A total of 1 416 differentially expressed genes were identified from GEO, while 5 646 CAVD-related targets were obtained from GeneCards. From HERB, Batman-TCM 2.0, and ETCM, 605 PNS targets were collected, with 130 overlapping genes identified. These were mainly involved in inflammatory responses, lipopolysaccharide-mediated signaling, transcriptional regulation, protein phosphorylation, and NF-κB pathway activation, occurring primarily in membrane rafts and the golgi apparatus, with molecular functions such as kinase activity regulation and protein binding, and enriched in lipid metabolism, atherosclerosis, and NF-κB signaling pathways. Experimental validation showed that PNS significantly decreased NF-κB p65 expression (P<0.05), significantly increased IκBα expression (P<0.01), and markedly suppressed expression of p-SYK (P<0.01), p-IκBα (P<0.01), p-NF-κB p65 (P<0.01), and p-NF-κB p50 (P<0.01). PNS also significantly reduced NF-κB p65 nuclear/cytoplasmic fluorescence intensity (P<0.01), decreased TNF-α secretion in the supernatant (P<0.01), and promoted IL-10 secretion (P<0.01). In conclusion, PNS ameliorates hVICs calcification by downregulating IκBα, NF-κB p65, and NF-κB p50 expression, inhibiting phosphorylation of SYK, IκBα, NF-κB p65, and NF-κB p50, reducing NF-κB p65 nuclear translocation, and suppressing TNF-α expression.

This study aims to investigate the effects of aged male parents on the learning ability of offspring and the intervention effect of Wuzi Yanzong Pills based on the microRNA-34a-5p (miR-34a-5p)/silent information regulator 1 (SIRT1) signaling pathway. Thirty-two SD male rats of 15 months old were randomized into aged model, model+high-dose (8 g·kg^-1) Wuzi Yanzong Pills, model+low-dose (2 g·kg^-1) Wuzi Yanzong Pills, and model+vitamin C (100 mg·kg^-1) groups (n=8). In addition, 8 SD male rats of 3 months old were selected as the control group. Rats in treatment groups were fed the diets containing different doses of Wuzi Yanzong Pills or vitamin C, and the control and model groups received a regular diet for 12 weeks. After 5 days of co-caging with 3-month-old female mice, the fertilization rate was recorded. An automated sperm analyzer was used to examine the sperm motility and count, and the testicular spermatogenesis was assessed by hematoxylin-eosin staining. The senescence cells in the testicular tissue was detected byβ-galactosidase staining, and miR-34a-5p expression was quantified via qPCR. The litter size was counted, and the body mass and body length were measured on days 1 and 30 to assess offspring development. For the offspring of 30 days old, their learning ability was examined via Morris water maze, and Nissl staining was employed to count hippocampal neurons. The miR-34a-5p expression in the hippocampal tissue of the offspring was determined by qPCR, and the protein levels of brain-derived neurotrophic factor (BDNF) and SIRT1 were determined by Western blot. Compared with the control group, the model group exhibited reductions in fertility rate, litter size, and sperm motility and count, as well as impaired testicular spermatogenesis (P<0.01). In addition, the model group showed increased senescence cells in testicular and epididymal tissue, accompanied by elevated miR-34a-5p expression in sperms. The 30-day-old offspring showed slow growth, reduced hippocampal neurons, up-regulated miR-34a-5p expression, and down-regulated protein levels of SIRT1 and BDNF in the hippocampus (P<0.01), along with impaired learning and memory performance (P<0.01). Compared with the model group, both high-dose Wuzi Yanzong Pills and vitamin C improved the fertilization rate, litter size, sperm motility, sperm count, and testicular spermatogenesis (P<0.05). The 30-day-old offspring in the two groups showed accelerated growth and development, increased hippocampal neurons, and elevated BDNF protein level in the hippocampus (P<0.05), along with enhanced learning and memory capabilities (P<0.05). Compared with the vitamin C group, the high-dose Wuzi Yanzong Pills group exhibited accelerated offspring growth (P<0.05), increases in fertilization rate and litter size (P<0.05), and improved learning and memory abilities (P<0.05). These findings indicate that Wuzi Yanzong Pills can improve testicular spermatogenesis and sperm quality in aged rats, thereby enhancing offspring's learning and memory performance. Specifically, Wuzi Yanzong Pills regulate miR-34a-5p expression to delay spermatogenic cell senescence in the testicular tissue and improve the offspring's cognitive function by miR-34a-5p mediated intergenerational transmission.

This study aims to explore the protective mechanism of Danqi Huayu Recipe against light-induced retinal damage (LIRD) in rats from the perspective of cell apoptosis. Sixty male SD rats were randomly allocated into six groups (n=10): normal group, model group, Ziyin Mingmu Pills group (7.8 g·kg^-1), and low-, medium-, and high-dose (3.45, 10.35, and 31.05 g·kg^-1, respectively) Danqi Huayu Recipe groups. A self-made light damage chamber was used for the modeling of LIRD in rats of other groups except normal group. Seven days after modeling, each group received corresponding agents by gavage. Normal group and model group were administrated with an equal volume of distilled water by gavage for 14 d. The retinal function was evaluated by electroretinography. Hematoxylin-eosin (HE) staining was employed to observe the retinal morphology and measure the thickness of the outer nuclear layer (ONL). The cell apoptosis was examined by terminal deoxynucleotidyl transferase dUTP nick end labeling. The ultrastructure of the retina was observed by transmission electron microscopy. The protein and mRNA levels of factors related to retinal cell apoptosis were determined by Western blot and RT-qPCR, respectively. The results showed that compared with normal group, model group demonstrated significantly decreased amplitudes of ERG a-waves and b-waves, thinned ONL, increased cell apoptosis rate, damaged retinal ultrastructure, down-regulated protein and mRNA levels of the anti-apoptotic factor (Bcl-2), and up-regulated protein and mRNA levels of pro-apoptotic factors (Caspase-3, Caspase-6, p53, and Bax). Compared with model group, Ziyin Mingmu Pills group and medium-and high-dose Danqi Huayu Recipe group recovered the amplitudes of ERG a-waves and b-waves, increased the ONL thickness, decreased the cell apoptosis rate, and improved the retinal ultrastructure. Meanwhile, they up-regulated protein and mRNA levels of Bcl-2 and down-regulated protein and mRNA levels of pro-apoptotic factors in a dose-dependent manner. In conclusion, Danqi Huayu Recipe protects rats from LIRD by regulating the expression of proteins and genes related to cell apoptosis to reduce light-induced retinal cell apoptosis.

This study aimed to investigate whether Xihuang Pills (XHP) enhance the therapeutic efficacy of temozolomide (TMZ) against glioblastoma (GBM) U251 cells by inducing mitochondria-associated ferroptosis via regulation of Nrf2/HO-1/GPX4 signaling axis. The XHP-containing serum was prepared, and the effects of various concentrations of TMZ (0-200 μmol·L^-1) and XHP (0-20%) on U251 cell viability were assessed by CCK-8 assay. The optimal synergistic dose (200 μmol·L^-1 TMZ + 10% XHP) was identified via the SynergyFinder platform for establishing a combined intervention model. Control, XHP, TMZ, and XHP + TMZ groups were designed. Cell proliferation was evaluated by EdU staining and colony formation assay, and cell migration and invasion were assessed by Transwell and wound healing assays. FerroOrange and DCFH-DA fluorescent probes were used to measure the intracellular levels of Fe²⁺ and ROS, respectively. The JC-1 fluorescent probe was used to assess mitochondrial membrane potential, while Mito-PeDPP fluorescent probe to examine mitochondrial lipid peroxidation. Mito-Tracker Green and MitoSOX^TM Red were used to examine mitochondrial morphology and oxidative stress. Transmission electron microscopy was adopted to observe the mitochondrial ultrastructure. GSH and MDA levels were measured by colorimetric assays. Western blot was employed to assess the protein levels of Nrf2, HO-1, xCT, and GPX4. The results showed that, XHP + TMZ suppressed cell viability, proliferation, migration, and invasion (P<0.01), increased Fe²⁺ accumulation, ROS generation, GSH depletion, and lipid peroxidation (P<0.001), reduced mitochondrial membrane potential, caused severe structural damage, and down-regulated the protein levels of Nrf2, HO-1, GPX4, and xCT, which indicated that the antioxidant ferroptosis-protective pathway was co-inhibited. In conclusion, XHP enhances TMZ-induced ferroptosis in GBM cells by suppressing the Nrf2/HO-1/GPX4 signaling axis. The findings provide new mechanism insights into the synergistic antitumor effect of traditional Chinese medicine in glioblastoma therapy and offer experimental evidence for the treatment of refractory brain tumors with combined strategies.

This study aims to compare the efficacy and safety of different traditional Chinese medicine injections (TCMIs) combined with conventional western medicine in the treatment of chronic obstructive pulmonary disease (COPD) with respiratory failure (RF) using a network Meta-analysis approach. A comprehensive literature search was conducted in CNKI, VIP, Wanfang, SinoMed, Web of Science, PubMed, Cochrane Library, and EMbase databases from inception to June 29, 2025. Randomized controlled trials (RCTs) were performed by employing the literature on the treatment of COPD with RF by TCMIs, and data were analyzed by Stata 17. A total of 110 RCTs involving 9 218 participants and 11 different TCMIs were included. The ranking of efficacy in the surface under the cumulative ranking curve (SUCRA) from network Meta-analysis was as follows: ① for improving clinical total response rate, Chuanxiongqin Injection + conventional western medicine, Shankezhi Injection + conventional western medicine, and Danhong Injection + conventional western medicine ranked top three; ② for reducing mortality, Chuanxiongqin Injection + conventional western medicine, Danshen Chuanxiongqin Injection + conventional western medicine, and Shenfu Injection + conventional western medicine ranked top three; ③ for shortening length of hospital stay, Danshen Chuanxiongqin Injection + conventional western medicine, Shenfu Injection + conventional western medicine, and Shenmai Injection + conventional western medicine ranked top three; ④ for reducing duration of mechanical ventilation, Shenqi Fuzheng Injection + conventional western medicine, Shenmai Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three; ⑤ for improving arterial partial pressure of oxygen (PaO₂), Chuanxiongqin Injection + conventional western medicine, Danshen Injection + conventional western medicine, and Danhong Injection + conventional western medicine ranked top three; ⑥ for reducing arterial partial pressure of carbon dioxide (PaCO₂), Shenfu Injection + conventional western medicine, Tanreqing Injection + conventional western medicine, and Danshen Chuanxiongqin Injection + conventional western medicine ranked top three; ⑦ for improving arterial oxygen saturation (SaO₂), Shenqi Fuzheng Injection + conventional western medicine, Xuebijing Injection + conventional western medicine, and Xingnaojing Injection +conventional western medicine ranked top three; ⑧ for improving forced expiratory volume in one second (FEV₁), Chuanxiongqin Injection + conventional western medicine, Shenmai Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three; ⑨ for improving forced vital capacity (FVC), Danshen Chuanxiongqin Injection + conventional western medicine, Xuebijing Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three. In terms of safety, adverse events were mostly mild and occurred at a low incidence. In conclusion, TCMIs combined with conventional western medicine may offer clinical benefits in the management of COPD with RF. However, due to limitations in the number and quality of studies for each intervention, further rigorously designed RCT should be conducted to validate these findings.