Helicobacter pylori (H. pylori) infects 48.5% of the global population, with approximately 42.8% in China and is linked to chronic gastritis and gastric cancer. Current mainstream antibiotic therapies against H. pylori are limited in clinical application due to antibiotic resistance and high recurrence rates. Consequently, there is an urgent need to develop novel therapeutic strategies. Research indicates that persistent H. pylori infection relies on its immune evasion mechanisms. Outer membrane vesicles (OMVs) serve as critical mediators in pathogen-host interactions, facilitating multiple immune escape strategies by delivering virulence factors, nucleic acids, and other substances. Deciphering the mechanisms of OMVs-mediated immune evasion in H. pylori is pivotal for overcoming the bottlenecks in OMVs-based H. pylori vaccine development and devising novel targeted therapeutic strategies. This review synthesizes recent research advances in OMVs-driven immune evasion, analyzes challenges in translational applications, and proposes future directions to inform innovative anti-H. pylori strategies.

Pruritus is an important physiological sensation mediated by the somatosensory system, and its clinical significance is increasingly recognized. Based on the underlying mechanisms, pruritus can be classified into histaminergic and non-histaminergic types. Non-histaminergic pruritus constitutes the major form of most chronic pruritus conditions and often shows poor response to conventional antihistamines. The gastrin-releasing peptide receptor (GRPR) in the spinal cord, serving as a pruritus-specific receptor, plays a critical role in the transmission of non-histaminergic pruritic signals. The κ-opioid receptor (KOR) and the μ-opioid receptor (MOR) exert opposite effects in pruritus regulation: MOR activation promotes pruritus, whereas KOR activation produces inhibitory effects. This review systematically elaborates on the multiple molecular mechanisms through which KOR inhibits pruritus, with a focus on the direct inhibition of GRPR function via protein kinase Cδ (PKCδ)-mediated phosphorylation, interference with intracellular calcium signaling, potential modulation of the phosphoinositide 3-kinase (PI3K)/Akt pathway, and comprehensive regulation of pruritic pathways through signaling molecules such as Gβγ and β-arrestin 2. Together, these mechanisms constitute the basis for KOR-mediated regulation of pruritic signals at the spinal level. Although KOR agonists (e.g., TRK-820, CR845) have shown therapeutic potential for chronic pruritus in clinical studies, translational challenges such as species differences and cross-talk among signaling pathways warrant further attention. In-depth elucidation of the KOR regulatory network not only provides novel insights into the physiological and pathological mechanisms of pruritus but also establishes a solid theoretical foundation for the development of novel, highly-targeted anti-pruritus drugs with fewer side effects.

Recent advances in understanding the modulatory functions of gut microbiota on human diseases facilitated our focused attention on the contribution of the gut to the pathophysiological alterations of many extraintestinal organs. Tryptophan-derived indole metabolites have become crucial signaling molecules in the gut-organ axis, playing a core role in anti-inflammation, immune homeostasis, and maintaining mitochondrial function by activating the aryl hydrocarbon receptor and pregnane X receptor. This review systematically explores the regulatory mechanisms of indole metabolites on host diseases through the gut-liver axis, gut-brain axis, gut-heart axis, and gut-lung axis, providing a new theoretical perspective for understanding the interaction between the gut microbiota and the host and developing new strategies for the prevention and treatment of related diseases.

Antibody-drug conjugates (ADC) are a type of innovative and highly effective anti-cancer drugs. They connect the high specificity of monoclonal antibody drugs with the efficient killing ability of small molecule toxins through a linker, effectively enhancing tumor targeting and reducing toxic side effects. As a highly promising innovative therapy in the field of cancer treatment, ADC have brought new hope to cancer patients. However, the issue of tumor permeability has become a key factor restricting the full exertion of their efficacy. This review introduces the mechanism of action and structural optimization strategies of ADC, analyzes the complex factors that hinder the permeation of ADC in the tumor microenvironment, and discusses the drug administration strategies to improve ADC permeability. Through the systematic review of relevant research progress, it provides theoretical basis and practical guidance for promoting the further development of ADC in tumor treatment.

Bacterial and parasitic infections rank as the second leading cause of death globally. The persistent rise in antibiotic resistance, particularly against β-lactams, vancomycinand methicillin, poses a significant challenge to public health. The World Health Organization (WHO) has prioritized the fight against antibiotic resistance, recognizing the urgent need for new targets and the development of novel antibiotics. Given that many potential drugs focus on pathogens prioritized by the WHO, which often share characteristics with existing antibiotic classes, fostering resistance, there is an imperative requirement to identify new targets and develop agents that specifically address these challenges. Bacterial peptidoglycan, with its specificity for bacteria and essential role in their survival, has become a focal point of research. Due to the enzymes involved in peptidoglycan biosynthesis being exclusive to bacteria, they represent ideal targets for the discovery of new antimicrobials. Inhibiting these enzymes offers an optimal inhibition profile, where the inhibitors are inactive towards eukaryotic cells but effectively combat bacterial infections. The aim of this review is to introduce the advancements in the synthesis of a series of inhibitors targeting the bacterial transferase MraY, which catalyzes the first membrane-bound step in peptidoglycan biosynthesis. Currently, there are no antibiotics in clinical use that target MraY. By inhibiting MraY, the hope is to mitigate the development of resistance. Natural inhibitors of MraY, based on the aminoarabinosyluridine scaffold, have been known for their biological activity. However, due to their limited antibacterial efficacy, several research groups have embarked on the development of simplified analogues to enhance their therapeutic potential.

QS-21 is an immunologically active saponin-based immunostimulant extracted from the bark of the South American soapbark tree (Quillaja saponaria, QS). It consists of four structural domains: Quillaic acid, a trisaccharide side chain, a tetrasaccharide chain, and a pseudodimeric acyl chain. It exists in two isomeric forms, QS-21-Api and QS-21-Xyl, among which the C-4 aldehyde group and the acyl side chain are the key moieties for adjuvant activity. In terms of its immunological mechanism, QS-21 can simultaneously activate Th1/Th2-type immune responses, enhance the cross-presentation of antigens by dendritic cells, and activate the MAPK (mitogen-activated protein kinase) pathway in T cells as well as the NLRP3 (NOD-like receptor protein 3) inflammasome. It exhibits particularly prominent performance in inducing cellular immunity. This article summarizes the currently developed composite adjuvant systems such as AS01, AS02, Matrix-M, AS15, and GLA-LSQ, along with their applications in vaccines including the malaria vaccine Mosquirix and the shingles vaccine Shingrix. Regarding issues such as supply shortages associated with natural extraction, technologies for chemical total synthesis, semi-synthesis, and yeast-based biosynthesis are making gradual breakthroughs, Strategies including some derivatives (such as VSA-1 and TQL1055) also provide references for the development of new vaccines and the prevention and control of infectious diseases as well as immunotherapy.

As indispensable medications in the medical field, antibiotics face severe challenges such as high pollution and high energy consumption in their traditional chemical synthesis processes. The novel enzymatic catalytic process, leveraging the catalytic properties of enzymes and the advantages of environmental friendliness, has emerged as a new direction for the green development of the antibiotic industry. This paper focuses on the application progress of the novel enzymatic catalytic process in the green synthesis of antibiotics, systematically elaborates on its catalytic mechanism, and analyzes the specific applications and advantages of this process in the production of antibiotics such as β-lactams, aminoglycosides, and macrolides. Meanwhile, it points out the issue that catalyst residues during the production process affect the quality and safety of pharmaceutical products, and discusses the current challenges confronted by the novel enzymatic catalytic process for antibiotics. Finally, by applying cutting-edge technologies including synthetic biology and artificial intelligence, this paper provides a theoretical basis and solutions for breaking through existing technical limitations and promoting the green upgrading of the antibiotic industry.

The current quality control standards for traditional Chinese medicine (TCM) are relatively simplistic, as relying solely on the quantification of chemical components fails to fully interpret the quality connotation of multi-effect medicinal substances. In contrast, phase-state analysis, which focuses on the physical forms of multiple components, is considered to provide a new perspective for researching the material basis of the efficacy of TCM. This article focuses on the challenges in quality control of multi-efficacy TCM, systematically analyzes the limitations of current standards in reflecting its multi-functional nature, and reviews research approaches and current progress on the role of physicochemical state in the material basis of TCM efficacy. It elaborates on the mechanism of self-assembly via non-covalent bonds and the structure-function synergy. Building on this, a new "top-down and bottom-up" integrated strategy for quality control research of multi-efficacy TCM is proposed. This aims to promote innovation in TCM quality control models, guide rational drug selection, and provide a scientific basis for constructing a quality evaluation system that aligns with the characteristics of TCM.

With the increasing severity of bacterial resistance issues, β-lactamase, as the key enzyme mediating resistance to β-lactam antibiotics, requires rapid and accurate detection that plays significant importance for clinical treatment and antimicrobial resistance surveillance. In recent years, the rapid development of proteomic technologies has provided novel strategies for high-throughput identification of β-lactamases. This article reviews the application progress of mass spectrometry-based proteomic technologies in β-lactamase detection, compared with traditional phenotypic detection and genetic sequencing methods, proteomic technologies enable direct capture of enzyme expression-level information, offering multidimensional data support for elucidating the association between resistant phenotypes and protein functions. Furthermore, advancements in targeted proteomics have enhanced detection sensitivity and specificity, facilitating the exploration of personalized anti-infective therapies. Looking forward, through the integration of artificial intelligence-assisted analysis and the development of miniaturized high-throughput detection devices, proteomic technologies are expected to play a more prominent role in real-time antimicrobial resistance monitoring and precision medicine.

In the tumor microenvironment (TME) of breast cancer, tumor-associated macrophages (TAMs) predominantly exhibit a pro-tumor M2 phenotype. These M2 TAMs significantly exacerbate breast cancer malignant progression through multiple mechanisms: they secrete growth factors and cytokines to promote the proliferation, invasion and migration of tumor cells; they facilitate tumor metastasis by inducing angiogenesis and remodeling the extracellular matrix; and they mediate the resistance of tumor cells to chemotherapy, targeted therapy and immunotherapy by providing survival signals, etc., leading to drug resistance. Consequently, TAMs are pivotal drivers of breast cancer initiation, progression, metastasis, and therapy resistance. With its excellent targeting, drug-carrying diversity and microenvironmental response properties, the nano-delivery system provides a new strategy for targeting and regulating the phenotypic polarization of TAMs by remodeling the inflammatory environment, modulating oxidative stress and immune signaling networks, and regulating the metabolic reprogramming of glucose, lipids, and amino acids, making it a key research direction for reversing TME, which has an important potential for clinical translation. This review systematically summarizes the critical impact of TAM phenotypic polarization on breast cancer pathogenesis, metastasis, and drug resistance, alongside nano delivery-based strategies for TAM reprogramming and the underlying molecular network mechanisms they regulate, aiming to provide new insights for the precision treatment of breast cancer.

The transdermal delivery system of traditional Chinese medicine (TCM) has emerged as a vital alternative to conventional administration routes, particularly for chronic disease management, owing to its advantages in avoiding hepatic first-pass effects, maintaining stable blood drug concentrations, improving patient compliance, and enabling sustained release. However, the efficiency of TCM transdermal delivery is significantly constrained by the "brick-and-mortar structure" barrier of the stratum corneum and the inherent complexity of TCM components, which also complicates quality control and mechanistic studies. To address these challenges, innovative strategies have been developed in recent years, including microneedles, nanocarriers (liposomes, nanoemulsions, polymeric nanoparticles, nanocrystals and self-assembled nanomicelles), permeation enhancers, iontophoresis, and sonophoresis. Synergistic permeation-enhancing strategies further overcome limitations of single-technique approaches. These technologies demonstrate substantial clinical value in dermatological therapy, pain management, wound healing, and related fields.

The elusive in vivo fate of nanomedicines represents a critical barrier to their clinical translation. Recent rapid advancements in imaging technologies, such as fluorescence imaging (FLI), photoacoustic imaging (PAI), radionuclide imaging (RI), magnetic resonance imaging (MRI), and multimodal imaging (MI) have provided powerful tools for real-time and in situ analysis of nanomedicine behavior in vivo, encompassing morphological evolution, drug release kinetics, distribution, metabolic and so on. However, significant challenges persist, particularly in continuously tracking dynamic morphological changes throughout their entire lifecycle and achieving accurate quantitative analysis within the physiological microenvironment. This review focuses on recent progress in understanding the in vivo fate of nanomedicines, systematically summarizing the fundamental principles, applications, advantages and limitations of these imaging technologies. It also highlights recent breakthroughs in utilizing these technologies to decipher the in vivo fate of nanomedicines and explores future perspectives, such as the development of innovative probes and the integration of multimodal imaging platforms. The objective is to offer robust theoretical and technical foundations for optimizing the design of nanodrug delivery systems and promoting their clinical translation.

Oral mucositis (OM) is a common complication of radiotherapy and chemotherapy in cancer patients. The severe pain and ulcer caused by OM not only significantly impair patients' quality of life but may also lead to the interruption of antitumor therapy. Currently, topical drug administration remains the preferred strategy for the prevention and treatment of OM. However, conventional dosage forms (e.g., mouthwash, gel) are susceptible to salivary clearance, exhibiting pharmacokinetic limitations such as short mucosal retention time and rapid drug clearance. In recent years, novel drug delivery systems, including nanocarrier, microneedle and hydrogel, have demonstrated significant advantages in enhancing mucosal drug targeting and prolonging therapeutic duration due to their excellent biocompatibility and controlled release properties. This review summarizes the recent research progress of the above three drug delivery systems in the prevention and treatment of OM, discusses current challenges faced by advanced delivery technologies, and outlines future research directions, aiming to provide insights for the development of more effective therapeutic strategies for OM.

Stem cell-like memory T cells (T_SCM), a subpopulation of T cells that combines features of both memory cells and stem cells, are capable of rapidly secreting immune cytokines and differentiating into effector T cells with tumor-killing ability after antigenic stimulation, thereby providing a lasting impetus for the anti-tumor immune response. Therefore, inducing T_SCM generation to bolster T cells' resistance to exhaustion, thereby improving tumor immunotherapy efficacy, has emerged as a major research focus. Nano-based drug delivery system (NDDS) possesses the qualities of in vivo targeted delivery and facilitated drug penetration, which provides the potential feasibility of inducing T_SCM strategy for clinical application. Based on this, this paper systematically describes the characteristics of T_SCM, reviews the current regulatory methods to induce and maintain T_SCM function, and summarizes the research progress of nano-based drug delivery system in regulating T_SCM for anti-tumor therapy, to provide reference and ideas for the application of induced T_SCM generation strategies in the clinic.

The gastrointestinal tract harbors the largest mucosal immune system, and tumor vaccines can leverage this advantage through oral administration to achieve enhanced immune responses. However, vaccines administered orally may encounter multiple physiological barriers that hinder their full efficacy. Nanocarriers can overcome these barriers. This paper discusses the physiological barriers currently faced by oral vaccines and the efforts made by nanotechnology to overcome these barriers. It also reviews the latest research progress and clinical translation status of nanoparticle-based delivery systems for oral tumor vaccines, aiming to provide research insights for the development of oral nanovaccines for the treatment of tumor-related diseases.

Gastrointestinal mucus constitutes a critical physiological barrier in the gastrointestinal tract. Its highly glycosylated mucin network effectively hinders the transport of particles and drug molecules toward the intestinal epithelium. In oral drug delivery, effectively overcoming this gastrointestinal mucus barrier is essential for achieving efficient drug absorption. On one hand, the continuously secreted mucus layer rapidly clears ineffectively penetrating particles due to its shear-thinning rheological property. On the other hand, macromolecular drugs face significant challenges in cross-mucus diffusion when their molecular size exceeds the mucus mesh pore size. Furthermore, certain small-molecule drugs can also be retained within the mucus layer through physicochemical interactions with its components, or rapidly eliminated by the dynamic turnover of the continuously secreted mucus, consequently leading to reduced drug delivery efficiency. Therefore, enhancing the mucus-penetrating capability of drug molecules is crucial for improving their oral bioavailability. This article systematically reviews the composition, structure, physicochemical properties, and barrier characteristics of gastrointestinal mucus, along with its barrier mechanisms affecting oral drug absorption. Additionally, it discusses nanotechnology-based drug delivery strategies designed to overcome the mucus barrier. The review aims to provide a theoretical basis and design principles for enhancing mucus permeability, improving oral bioavailability, and developing highly efficient mucus-penetrating nano-formulations.

Conventional antibody discovery paradigms mainly rely on animal immunization and hybridoma screening, which results in long development lead times, high costs, and a low success rate for antibody therapeutics. Recently, artificial intelligence (AI) is constantly driving the progress of antibody drug research and development technology with advantages in large-scale proteomic data processing. It plays a great important role in the prediction of antibody structure and de novo generation of target-specific sequences, which also substantially enhances the hit rate of drug candidates. Therefore, this article aims to summarize recent developments and challenges in the application of AI technology to antibody discovery and design, while also considering the limitations to traditional methodologies. Hope to provide practical guidance for AI-driven antibody drug development.

This study aims to investigate the inhibitory effects of cryptotanshinone (CPT) on colorectal cancer (CRC) both in vitro and in vivo, and to explore its potential mechanism of action based on ferroptosis. CRC HCT116 and HCT8 cells were treated with different concentrations of CPT. Cell counting, colony formation assays, and Annexin V/PI staining revealed that CPT significantly inhibited CRC cell viability and proliferation and promoted apoptosis. A CRC patient-derived organoid (PDO) (in accordance with the regulations of the Nankai University Biomedical Ethics Committee approval number NKUIRB2025036) model was established, and cell counting, hematoxylin-eosin (HE) staining, and immunohistochemistry (IHC) detection of the tumor marker Ki67 confirmed that CPT suppressed PDO growth and malignant proliferation. A mouse xenograft tumor model was established (in compliance with the regulations of the Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine, approval number: TCM-LAEC2025259K2011). Measurements of tumor volume and weight, HE staining, and Ki67 IHC demonstrated that CPT significantly inhibited tumor growth in vivo. Further studies found that a ferroptosis inhibitor could reverse the inhibitory effect of CPT on CRC cell proliferation, and CPT showed synergistic effects with ferroptosis inducers. Western blot and other detection results showed that CPT reduced the ratio of intracellular reduced glutathione (GSH) to oxidized glutathione (GSSG), and increased the levels of 4-hydroxynonenal (4-HNE) and the lipid peroxidation product malondialdehyde (MDA). Mechanistic studies indicated that CPT decreased glutathione peroxidase 4 (GPX4) mRNA levels and protein expression in a concentration-dependent manner. In conclusion, CPT can induce ferroptosis in CRC cells through GPX4, thereby inhibiting CRC cell proliferation and exerting anti-colorectal cancer effects.

In this study, the formation mechanism of the nanophase of "Rhizoma Anemarrhenae-Gypsum" was investigated, and its compatibility, antipyretic efficacy, and underlying mechanisms were systematically evaluated via network pharmacology. Three groups, namely the Rhizoma Anemarrhenae group, the Gypsum group, and the Rhizoma Anemarrhenae-Gypsum group, were set, with all groups including both nanophase preparations and traditional decoctions. The particle size, salinity, electrical conductivity and surface tension of the solutions were determined, and these combined with transmission electron microscope (TEM) and fingerprint analysis. Targets were screened using the GeneCards and DisGeNet, an active component-target network was constructed, a PPI network was built with the help of the STRING database and Cytoscape, and GO and KEGG enrichment analysis was performed using Metascape. The antipyretic effect of the nanophase solution was verified by hematoxylin-eosin (HE) staining, Western blot and real-time PCR. The results indicated that salinity and electrical conductivity promoted the stable formation of the nanophase; and TEM revealed that timosaponin coated the Gypsum crystal structure, leading to nanoparticle formation. Fingerprint analysis identified eight common peaks (neomangiferin and mangiferin as the main components), and the chemical components of the nanophase were consistent with those of the decoction. Through network pharmacology, 31 active components of Rhizoma Anemarrhenae and Gypsum were obtained, corresponding to 138 potential targets; the number of fever-related targets was of 1 093 fever-related targets were retrieved, among which 188 overlapping targets were screened. The main KEGG enrichment pathways included epidermal growth factor receptor, interleukin-17, hypoxia-inducible factor 1 and other signaling pathways. Experimental results showed that the nanophase group of Rhizoma Anemarrhenae-Gypsum decoction significantly reduced the body temperature in febrile rats, inhibited the expression of inflammatory factors, down-regulated fever medium, up-regulated cytoprotective proteins, and suppressed the expression of related mRNA. These findings were consistent with the predictions of network pharmacology. All animal experiments in this study were approved by the Animal Experimental Ethics Committee of Harbin University of Commerce (HSDYXY-2024022). This study found that Rhizoma Anemarrhenae-Gypsum enhances its therapeutic efficacy through the nanophase, and its antipyretic mechanism involves the multi-target regulation of inflammatory pathways and central signaling molecules thereby providing both theoretical and experimental support for the modernization of traditional Chinese medicine compound prescriptions.

Mesenchymal stem cells (MSCs) and human umbilical cord MSC-derived exosomes (hUC-MSCs-Exos) based therapy has been widely regarded as a potential strategy for the treatment of hepatic fibrosis, and the aim of the present study was to investigate the effects of human umbilical cord MSCs (hUC-MSCs) and hUC-MSCs-Exos on hepatic fibrosis and to reveal their potential mechanisms of action. By constructing an animal model of carbon tetrachloride (CCl₄)-induced hepatic fibrosis, observing the histomorphological and pathological changes, and combining the changes in liver function indices, inflammatory factor levels and peroxide content, we comprehensively evaluated the anti-fibrotic ability of hUC-MSCs and hUC-MSCs-Exos. Meanwhile, miRNA sequencing (miRNA-seq) technology was used to screen for the differentially expressed genes, and the role of hUC-MSCs and hUC-MSCs-Exos on the regulation of proteins related to the Wnt/β-catenin signaling pathway was verified with the help of protein immunoblotting (Western blot, WB). All protocols and procedures were approved by the Animal Ethics Committee of the Animal Experiment Center of Xinjiang Medical University, and all experimental procedures complied with Chinese regulations concerning laboratory animals (approval No.: IACUC-JT-20240627-26). The results showed that both hUC-MSCs and hUC-MSCs-Exos could effectively alleviate hepatic fibrosis by reducing collagen accumulation, improving liver function, inhibiting inflammation, and decreasing oxidative stress, and hUC-MSCs-Exos was more effective than hUC-MSCs in treating hepatic fibrosis; at the same time, both of them could down-regulate the expression of Wnt/β-catenin pathway components (Wnt3a, β-catenin, WISP1/CCN4, cyclin D1) in the hepatic fibrosis tissues. In addition, miRNA-seq results further indicated that the expression of miRNA-184 was significantly involved in the regulation of this pathway. The findings in this paper confirmed that both hUC-MSCs and hUC-MSCs-Exos improved liver fibrosis in rats, and the antifibrotic effect of hUC-MSCs-Exos may be related to the Wnt/β-catenin signaling pathway, which provides an effective therapeutic strategy for the treatment of liver fibrosis.

The purpose of this study was to investigate the potential regulatory mechanisms underlying the neuroprotective effects of Gualou Guizhi Decoction (GLGZD) in cerebral ischemia-reperfusion (CI/R)-injured rats. The main chemical components of GLGZD were identified by high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOFMS). Molecular docking was performed using the CB-Dock2 platform to evaluate the binding interactions between GLGZD components and Fn14. After the middle cerebral artery occlusion (MCAO) model was established in male SD rats, the therapeutic efficacy of GLGZD was evaluated using a combination of assessments. The modified neurological severity score (mNSS) and Catwalk test were conducted to evaluate neurological and motor functions of rats. MRI was used to measure cerebral infarction volume. Nissl staining was performed to observe the morphological structures of the neurons. Western blot was used to detect protein expression of tumor necrosis factor-like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible immediate-early response protein 14 (Fn14), TNF receptor associated factor 2 (TRAF2), cellular inhibitor of apoptosis 1 (cIAP1), NF-κB inducing kinase (NIK), inhibitor of kappa B kinase α (IKKα), phosphorylated IKKα, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) in ischemic cortex. Immunofluorescence was used to analyze the expression of Fn14 in neurons, and immunohistochemistry was employed to examine the nuclear translocation of nuclear factor kappa B (NF-κB). The results showed that the 8 identified ingredients of GLGZD could bind to Fn14. Compared with the sham surgery group, the model group exhibited significantly reduced motor function, Nissl-positive cells, and protein expression levels of cIAP1, TRAF2, and total Bcl-2 (P < 0.01). Conversely, mNSS scores, cerebral infarction volume, TWEAK, Fn14, NIK, Bax protein expression levels, IKKα phosphorylation levels, and neuronal Fn14-positive cell counts all significantly increased (P < 0.01). GLGZD treatment markedly improved motor function in MCAO rats, increased the number of Nissl-positive cells in the ischemic cortex, and enhanced cIAP1, TRAF2, and total Bcl-2 protein expression (P < 0.05, P < 0.01). It also significantly reduced mNSS and cerebral infarction volume, decreased TWEAK, Fn14, NIK, and Bax protein expression in the ischemic cortex, lowered IKKα phosphorylation, and reduced neuronal Fn14-positive cell counts (P < 0.05, P < 0.01). In conclusion, GLGZD can improve neurological and motor function after stroke in MCAO rats. This neuroprotective effect may be mediated through the TWEAK/Fn14/NF-κB pathway, which reduces neuronal damage. The animal study was approved by the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine (approval number: FJTCM IACUC 2021087).

This study aimed to investigate the interventional effect and potential mechanism of a lignan compound, (7S, 8R, 8'R)-eplignan C (EPC), from Ephedra intermedia, on ovalbumin (OVA)-sensitized asthmatic mice. Specific pathogen-free (SPF) male BALB/c mice were randomly assigned to normal control (CON), model (M), positive control (montelukast sodium, 1.3 mg·kg^-1), EPC low-dose (EPC-L, 10 mg·kg^-1), and EPC high-dose (EPC-H, 20 mg·kg^-1) groups. An allergic asthma model was established through intraperitoneal injection of OVA with aluminum hydroxide adjuvant followed by OVA aerosol challenge. After 7 consecutive days of administration, cough and asthma indicators, lung function, serum immunoglobulin E (IgE) levels, and lung tissue pathology were assessed 24 h post-final dose. Enzyme-linked immunosorbent assay (ELISA) was employed to measure serum levels of mast cell degranulation products and inflammatory factors; flow cytometry was utilized to detect immune cells (dendritic cells, mast cells, Tregs) and Ca²⁺ levels; immunofluorescence was applied to evaluate tryptase alpha/beta 1 (TPSAB1) and septin 11 (Septin11) protein expression in lung tissue; Western blot analysis was conducted to examine protein expression and activation levels of key molecules in the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in lung tissue samples. Furthermore, a co-culture system comprising mouse lung epithelial cells (MLE-12) and primary mouse splenocytes, combined with the JAK2/STAT3-specific inhibitor tyrphostin AG490, was employed to elucidate the mechanism underlying EPC's intervention in OVA-sensitized asthma. Results demonstrated that EPC alleviated asthma-related cough, inhibited airway hyperresponsiveness, improved lung pathological damage, modulated immune cell levels, suppressed inflammatory responses and mast cell degranulation, exhibiting significant anti-asthmatic effects, and reduced key protein expression in the JAK2/STAT3 signaling pathway. Further mechanistic investigation revealed that EPC regulated Th17/Treg immune imbalance and attenuated inflammatory responses in the co-culture system, effects that were blocked by AG490. Thus, it is speculated that EPC may exert its anti-asthmatic effects by regulating the JAK2/STAT3 signaling pathway to inhibit mast cell degranulation and modulate immune-inflammatory responses. This animal experiment gas been approved by the Animal Ethics Committe of Henan University of Chinese Medicine (approval number DWLL2018080003).

The aim of this study is to elucidate the mechanism of metadoxine through which it attenuates alcoholic liver disease-associated hepatic steatosis. In this study, the effect of metadoxine on alcoholic liver disease was evaluated in mouse model of alcoholic liver disease induced by high alcohol and high fat diets. In addition, RNA sequencing (RNA-Seq) of the liver tissues was applied to determine the dominant signaling pathways in response to metadoxine treatment. The mechanism of metadoxine on metabolic was investigated by mitochondrial function in HepG2 cells. The animal experiments in this study were conducted in accordance with the guidelines of the Ethics Committee for Scientific Research of Hefei University of Technology and were approved by the Animal Experiment Ethics Committee (approval No.: HUFT20210615002). When mice were fed a diet designed to induce alcoholic liver disease, hepatic lipid accumulation resulted. In this study, the level of serum biomarkers of liver injury, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST), was substantially decreased in mice treated with metadoxine compared to the model group (P < 0.05). Meanwhile, the inflammatory infiltration and steatosis in liver were alleviated by the administration of metadoxine compared to the model group (P < 0.01). In vitro, metadoxine significantly reversed ethanol overload-induced impairment of mitochondrial respiratory function and increased mitochondrial membrane potential in hepatocytes. Transcriptomic sequencing (RNA-Seq) analysis of liver tissue identified significant enrichment of the mitogen-activated protein kinase (MAPK) signaling pathway. Measurement of MAPK phosphorylation levels in liver tissue showed that metadoxine treatment significantly reduced MAPK phosphorylation (P < 0.01). Furthermore, the protective effects of metadoxine were abolished upon co-treatment with a p38-MAPK inhibitor. Conversely, co-treatment with a p38-MAPK activator reversed the inhibitory effect of metadoxine on p38-MAPK phosphorylation. This study investigated the mechanism of metadoxine in alcoholic liver disease and found that metadoxine enhanced mitochondrial function by the phosphorylation of MAPK and reduced the lipid accumulation and inflammation in alcoholic liver disease.

To investigate the regulatory effects of icariin (ICA) on the proliferation of H9C2 cardiomyocytes injured by oxygen and glucose deprivation/reperfusion (OGD/R) and on filamin B (FLNB)/β-actin (ACTB)/cyclin D1 (CCND1) signaling pathway, H9C2 cardiomyocytes were used as the research model. Cell viability was determined using the cell counting kit-8 assay. The cells were divided into three groups, normal control group, OGD/R group, and ICA group. The normal control group was cultured under normal conditions, while the other two groups were subjected to 4 hours of OGD followed by 2 hours of reperfusion. Enzyme-linked immunosorbent assay was employed to measure the levels of aspartate transaminase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK). Transcriptome sequencing was performed to identify the target genes of ICA in H9C2 cells; differentially expressed genes (DEGs) were analyzed via gene ontology (GO) functional annotation and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment. Molecular docking and molecular dynamics simulation were used to evaluate the binding affinity between ICA and core targets. Compared with the OGD/R group, ICA groups at concentrations of 1-10 μmol·L^-1 exerted a significant protective effect, and the levels of AST, LDH, and CK were notably decreased. Transcriptome sequencing results revealed that ICA significantly regulated 436 common DEGs in H9C2 cells after OGD/R, primarily affecting biological functions such as anatomical structure morphogenesis, protein binding, and intracellular anatomical structure, as well as signaling pathways including focal adhesion and pathways in cancer. STRING database analysis identified core targets including FLNB, ACTB, and CCND1. ICA significantly reversed the OGD/R-induced upregulation of FLNB, ACTB, and CCND1 expression. Molecular docking demonstrated strong binding between ICA and the core differential proteins, and molecular dynamics simulation confirmed that ICA significantly maintained the stability of FLNB protein. In conclusion, ICA can alleviate OGD/R-induced injury in H9C2, reduce the levels of AST, LDH, and CK, and downregulate the expression of FLNB, ACTB, and CCND1. This protective effect may be closely associated with ICA's binding to FLNB directly and subsequent mediation of the ACTB/CCND1 signaling pathway.

The pyrogallol compounds containing adamantanes were designed and synthesized by acyl chloride method and carbodiimide condensation method in order to obtain better anti-coronavirus activity compounds. All the 20 target compounds were characterized by mass spectrometry, ¹H NMR and ¹³C NMR, and the coronavirus main protease (Mpro) inhibitory activity and molecular docking were tested. The activity of 20 pyrogallol analogues targeting Mpro was screened. The IC₅₀ values of compounds 2e and 2m were 0.98 and 0.36 μmol·L^-1, respectively. Compound 2e is close to the positive drug (baicalein) IC₅₀. Compared with baicalein, compound 2m has better activity, and its IC₅₀ value is about 0.4 times that of baicalein. The molecular docking results show that it interacts with SARS-CoV-2 Mpro mainly through hydrogen bonds and hydrophobic bonds. The hit compound 2m obtained in this study provides a new material basis and research ideas for the development of anti-coronavirus drugs.

β-Lactamase inhibitor sulbactam has unique activity against Acinetobacter baumannii, but its direct target protein has not been fully elucidated. In this research, thermal proteome profiling and surface plasmon resonance technologies were applied to identify and validate RNA 5′ pyrophosphohydrolase (RppH) as the direct target of sulbactam against A. baumannii. And further, using the checkerboard method, we found that sulbactam could synergize with carbapenem antibiotics to battle A. baumannii. The research results supplement the new molecular mechanism of action of sulbactam and provide an effective combined regimen for its combination with other antibiotics to combat clinical intractable drug-resistant bacteria infections.

Based on the ProTide technology, structural modification was performed on the 3′-hydroxyl group of 2′-deoxy-5-fluorouridine (FdUR) to design and synthesize a series of novel 5-FU phosphoramidate diester dual prodrugs (A1-A10). The structures of all compounds were confirmed by spectroscopic techniques, and their anti-hepatocellular carcinoma activity was evaluated using the MTT assay. The results indicated that several compounds exhibited significant anti-liver tumor activity. Among them, compound A6 showed the most potent activity with IC₅₀ values of 0.15 μmol·L^-1 (against HepG2 cells) and 0.34 μmol·L^-1 (against Huh7 cells), which were superior to the positive control 11b. It also demonstrated good selectivity towards normal cells. Mechanistic studies revealed that ³¹P NMR monitoring confirmed that A6 follows the classic ProTide activation pathway and can be hydrolyzed by carboxypeptidase Y to remove the carboxylate ester. In vitro metabolic stability studies indicated that A6 remained stable in rat plasma for up to 48 h, whereas it underwent rapid activation in liver homogenate. Further in vivo experiments confirmed that in a mouse orthotopic Huh7 xenograft model, A6 (45 mg·kg^-1, administered every 2 days) achieved a tumor inhibition rate of 55.84%, which was significantly higher than that of the sorafenib control group (38.98%), without causing significant changes in body weight. The animal protocols were approved by the Animal Ethics Committee of Capital Medical University (No. AEEI-2025-536). This study successfully developed a class of 5-FU dual prodrug candidates with significant anti-HCC activity. Among them, A6 exhibited excellent in vitro activity, metabolic stability, and in vivo antitumor efficacy. Its activation mechanism aligned with the designed pathway, providing a valuable lead compound and theoretical support for anti-HCC drug development.

To further clarify the neuroprotective active components of the marine-derived fungus Phaeosphaeriopsis sp. WP-26, the extract from its rice medium fermentation product was isolated and purified using various column chromatography techniques and high-performance liquid chromatography (HPLC). Nine compounds, including 5 isocoumarin monomers and 4 isocoumarin dimers, were isolated from the extract of the fermentation product of Phaeosphaeriopsis sp. WP-26. The chemical structures of the compounds were elucidated via NMR experiments, high-resolution mass spectrometry analysis, X-ray diffraction analysis, and comparison with the literature. Their structures were identified as phaeosphaerin F (1), 4-chloro-6, 8-dihydroxy-3-methylisochroman-1-one (2), 6-demethylkigelin (3), (3R)-6, 7-dimethoxymellein (4), lignicol (5), spegazmarin C (6), spegazmarin D (7), spegazmarin E (8), and spegazmarin F (9). Among them, compound 1 is a new isocoumarin derivative, and compounds 2, 6-9 were isolated from fungi of the genus Phaeosphaeriopsis for the first time. Compounds 1-9 exhibited neuroprotective effects to varying degrees against H₂O₂-induced SH-SY5Y cell damage.

Cyanobacteria are an important source of novel chemical structures and bioactive lead compounds. A new anabaenopeptin, okeanopeptin A (1), was isolated from the culture products of a new genus of marine cyanobacterium Okeanomitos corallinicola. Its planar structure and absolute configuration were elucidated through extensive spectroscopic analysis including NMR and HRESIMS, as well as chemical derivatization. This compound represents the first reported anabaenopeptin incorporating an N-methylphenylalanine residue. Biological evaluation revealed that okeanopeptin A exhibited weak inhibitory activity against carboxypeptidase A.

Through several chromatographic methods, such as silica gel, Sephadex LH-20, medium-pressure liquid chromatography (MPLC), combined with modern spectroscopic techniques including nuclear magnetic resonance (1D and 2D NMR) and high resolution mass spectrometry (HRMS), two iridoids and six oleanane-type triterpenoids were isolated and identified from the roots and rhizomes of Patrinia scabiosaefolia. Among them, compound 1 was a new 3, 8-epoxy iridoid and named as patrinin C (1). Compounds 2-8 were isolated from this plant for the first time, and their structures were identified as alboside II (2), 11α, 12α-epoxy-3β, 23-dihydroxyolean-28, 13β-olide (3), 23-hydroxy-3-oxo-28-nor-12, 16-dien oleanane (4), 3′-O-acetyl-3-O-α-L-arabinosyl-23-hydroxyolean-12-en-28-oic acid (5), 2′, 3′-O-diacetyl-3-O-α-L-arabinopyranosyl-23-hydroxyolea-12-en-28-oic acid (6), hederagenin 3-O-(2-O-acetyl-β-D-xylopyranosyl)-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside (7) and pictoside A (8), respectively.

By comprehensively applying various separation techniques such as silica gel, ODS, and Sephadex LH-20 column chromatography, as well as preparative HPLC, fourteen C21 steroidal compounds were isolated from the chloroform extract of Cynanchum wallichii Wight. The structures of these compounds were identified through spectroscopic techniques including mass spectrometry (MS), nuclear magnetic resonance spectroscopy (NMR), ultraviolet spectroscopy (UV), and infrared spectroscopy (IR) methods as cynanotonin E (1), qingyangshengenin (2), qinyangshengenin-3-O-β-D-cymaropyranoside-(1→4)-β-D-cymaropyranoside (3), qinyangshengenin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosy-(1→4)-β-D-digitoxopyranoside (4), qinyangshengenin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside (5), 12-O-benzoylsarcostin-3-O-β-D-oleandropyranosy-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl (6), 12-O-benzoylsarcostin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranosyl (7), caudatin-3-O-β-D-digitoxopyranoside (8), caudatin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranoside (9), caudatin-3-O-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside (10), caudatin-3-O-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside (11), otophylloside B (12), otophylloside E (13) and otophylloside F (14). Among them, compound 1 is a new compound. The cytotoxic activities of compounds 1-14 against human hepatocellular carcinoma HepG2 cells were tested using the MTS method. The results indicated that all compounds exhibited good inhibitory activities against the proliferation of HepG2 cells, with IC₅₀ values ranging from 4.3 ± 1.25 - 26.3 ± 1.65 μmol·L^-1. Among them, the inhibitory activities of compounds 1, 3 and 6-14 were all stronger than that of the positive control cisplatin (DDP, IC₅₀: 15.9 ± 1.25 μmol·L^-1).

The chemical constituents in the bark of Eucommia ulmoides were isolated and purified using chromatographic techniques, including silica gel, Sephadex LH-20, ODS, MCI, and semi-preparative HPLC. The structures of the compounds were elucidated by spectroscopic analyses (NMR, IR, UV, and MS). A total of six compounds were identified as duzhonglactone (1), (7S, 8R)-4-hydroxy-3, 3′, 5′-trimethoxy-8, 9-dinor-8, 4′-oxyneoligna-7, 9-diol-7′-aldehyde (2), armaosigenin (3), (-) evofolin B (4), N-trans-feruloyltyramine (5), and C-veratroylglycol (6). Among them, compound 1 was a new five-membered ring lactone, while compounds 2-5 were isolated for the first time from Eucommia ulmoides bark. Furthermore, compounds 2-4 demonstrated antioxidant activity, with IC₅₀ values for 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging of 46.34 ± 5.80, 74.17 ± 8.71, and 78.02 ± 8.94 μg·mL^-1, and for 2, 2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging of 50.56 ± 1.35, 54.37 ± 3.04, and 55.61 ± 3.41 μg·mL^-1, respectively.

Through repeated application of various chromatographic techniques including silica gel, ODS, MCI, and Sephadex LH-20, 15 compounds were isolated from Magnolia officinalis var. biloba in Lishui. The structures and relative configurations of these compounds were determined using spectroscopic methods such as high-resolution electrospray ionization mass spectroscopy (HRESIMS) and 1D & 2D-NMR. The absolute configurations of compounds (+)-1, (-)-1, (+)-5, and (-)-5 were ultimately established through chiral resolution techniques and quantum chemical calculations of electronic circular dichroism (ECD) spectra. Among these, compounds (+)-1, (-)-1, 2-4, (+)-5, (-)-5, and 6-9 were previously unreported neolignans primarily derived from honokiol or magnolol, while compounds 10-13 were known compounds. The anti-inflammatory activities of all compounds were evaluated using an LPS-induced RAW264.7 cell model. Among them, compound 4 exhibited significant inhibitory activity against nitric oxide (NO) production, with an IC₅₀ value of 1.23 ± 0.04 μmol·L^-1.

In recent years, the synthetic cannabinoid N-(1-amino-3, 3-dimethyl-1-oxobutan-2-yl)-1-butyl-1H-indazole-3-carboxamide (ADB-BUTINACA) has attracted widespread attention due to its high affinity for the CB1 receptor and associated abuse risk; however, its toxicity mechanism remains poorly understood. This study aimed to systematically investigate the toxicological mechanisms of ADB-BUTINACA using an integrated approach. We established an acute rat exposure model (low and high doses: 0.1 and 1 mg·kg^-1) and employed untargeted metabolomics combined with molecular docking analysis to examine its effects on the urinary metabolome and toxicity pathways. Our analysis identified 52 significantly altered metabolites, including taurine and phenylacetaldehyde, and revealed six disturbed metabolic pathways, such as phenylalanine metabolism and one-carbon metabolism. Notably, choline and methionine were pinpointed as potential dose-dependent biomarkers. Molecular docking results demonstrated a strong binding affinity (-7.8 kcal·mol^-1) between ADB-BUTINACA and aldehyde dehydrogenase ALDH3, suggesting that the compound inhibits ALDH3 activity. This inhibition likely disrupts phenylalanine metabolism, leading to the accumulation of phenylacetaldehyde and the upregulation of phenylacetylglycine, thereby inducing oxidative stress and potential cardiovascular and neurotoxicity. Concurrently, the downregulation of methionine and choline indicated an impairment of one-carbon metabolism, which may further exacerbate oxidative stress. In conclusion, this study elucidates the mechanism by which ADB-BUTINACA exposure induces oxidative stress primarily through disrupting phenylalanine and one-carbon metabolism, utilizing non-targeted metabolomics and molecular docking. The identified biomarkers provide a crucial theoretical foundation for future research on the toxicity mechanisms, clinical detection, and risk assessment of synthetic cannabinoids. The experimental protocol complied with the welfare and ethical requirements for experimental animals and was approved by the Animal Ethics Committee of Zhejiang Chinese Medical University (Approval number: 20220718-11).

Yimaikang Soft Capsule is a traditional Chinese patent medicine used to promote blood circulation, remove blood stasis and improve microcirculation in the treatment of ischemic cardio-cerebrovascular diseases. However, its chemical composition and mechanism of action remain unclear. In this study, the chemical constituents of Yimaikang Soft Capsules were systematically analyzed using high-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (HPLC-Q-Exactive Orbitrap MS) and global natural products social molecular networking (GNPS). A total of 125 in vitro components were identified, including 12 compounds reported for the first time in the Yimaikang series of preparations and its raw material herb Erigeron breviscapus. Subsequently, network pharmacology was employed to predict and screen 160 common targets shared between the 125 identified components and the diseases (ischemic cerebrovascular disease and coronary heart disease). Based on these, a "ingredient-target" interaction network and a protein-protein interaction (PPI) network were constructed to identify the core components and key targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the mechanism of Yimaikang Soft Capsules primarily involves signaling pathways such as the vascular endothelial growth factor (VEGF) signaling pathway, epidermal growth factor receptor (EGFR) signaling pathway and the advanced glycation end-product receptor (AGE-RAGE) signaling pathway. These pathways are closely associated with biological processes including angiogenesis, inflammation regulation, and apoptosis. Molecular docking validation demonstrated strong binding activity between core components (such as 3, 5, 6, 4′-tetrahydroxy-7-methoxyflavone, aesculin and kaempferol) and their corresponding targets (phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit beta, PIK3CB; Harvey rat sarcoma viral oncogene homolog, HRAS; phosphoinositide-3-kinase regulatory subunit 1, PIK3R1). This study preliminarily identified the potential active ingredient group of Yimaikang Soft Capsules and elucidated its multi-target, multi-pathway mechanism of action, providing a scientific basis for further clarifying the modern pharmacological foundation of this formulation.

Rapid and accurate identification of chemical components in classical famous formulas is a crucial aspect of their modern research. This study aimed to establish a method for the rapid identification of chemical components in the mass spectrometry data of Xie Bai San by integrating multi-point mass defect filtering (MDF) and molecular networking (MN) techniques, to achieve accurate and comprehensive characterization of its constituents. The mass spectrometric data of Xie Bai San decoction was acquired using ultra-high performance liquid chromatography coupled with quadrupole-Orbitrap tandem mass spectrometry (UHPLC-Q-Orbitrap-MS/MS) in both positive and negative ion modes. The MDF method was employed, utilizing set multi-point mass defect filtering windows to identify structurally similar compounds. The MN approach was applied to create a GNPS molecular network based on the similarity of MS/MS fragmentation patterns, and Cytoscape 3.6.1 software was used to identify clusters of structurally similar molecules, enabling the identification of homologous components via probe molecules. Using the aforementioned methods, combined with retention time, accurate molecular mass, and secondary MS/MS fragmentation characteristics, and based on database and relevant literature information, a total of 185 compounds were identified in the Xie Bai San decoction. These included 54 flavonoids, 42 alkaloids, 17 saponins, 16 stilbenes, 34 organic acids, 15 phenylpropanoids, and 7 other types of compounds. Among these, 46 compounds were reported for the first time in the Xie Bai San formula or its constituent herbs. This study successfully achieved the rapid, accurate, and comprehensive characterization of the chemical components in Xie Bai San decoction. It provides a chemical component foundation for research on the pharmacodynamic material basis and quality control of Xie Bai San compound preparations, and also offers a methodological reference for the rapid identification of chemical components in complex traditional Chinese medicine systems.

This study developed a sustained-release lipid liquid crystalline (LLC) system for leuprolide acetate (LA) and investigated the feasibility of using the lipase inhibitor orlistat (Orl) to modulate the matrix degradation and prolong the drug release period. The LA-LLC formulation was characterized by using polarized light microscopy (PLM), rheological analysis, and in vitro release studies to guide optimization. The impact of Orl on the sustained-release performance of LA-LLC was evaluated through in vitro degradation, in vitro release, and pharmacokinetic studies in rats. All animal protocols were approved by the Laboratory Animals Care and Use Committee of Yantai University (registration number is YTU20210319) (Yantai, China). Results showed that the LLC system utilizing glycerol dioleate (GDO) as the oil phase exhibited optimal sustained-release characteristics. The optimized LA-LLC formulation comprised soybean phosphatidylcholine, GDO, ethanol, and LA at a mass ratio of 44.5∶44.5∶10∶1. This gel exhibited a hexagonal phase structure by PLM. It demonstrated shear-thinning capacity characteristic of a pseudoplastic fluid. In vitro degradation and release results indicated that Orl significantly inhibited lipid degradation and prolonged drug release. Pharmacokinetic results in rats indicated that the addition of inhibitor Orl prolonged the release period from 42 days to 56 days compared to the LA-LLC group. This study demonstrates that the LA-LLC system, with its structural stability and controllable release, can effectively prolong drug release by employing a lipase inhibitor to regulate matrix degradation. The results of the study provide a novel strategy for the sustained delivery of peptide drugs.

Helicide and rosmarinic acid possess sleep-improving properties but suffer from poor aqueous solubility. This study aimed to construct a dual-drug co-loaded amorphous system. Compatibility and intermolecular interactions were predicted using Hansen solubility parameters and molecular docking. Co-evaporates were prepared via the solvent co-evaporation method. Polarized light microscopy, X-ray diffraction, and differential scanning calorimetry analyses confirmed the formation of a single-phase binary co-amorphous system. Fourier-transform infrared spectroscopy, ultraviolet spectroscopy, and nuclear magnetic resonance analyses revealed that hydrogen bonds formed between the aromatic ketone carbonyl group of helicide and the carboxylic acid group of rosmarinic acid, while π-π stacking interactions occurred between their aromatic rings, constituting a stable intermolecular network. Co-amorphization significantly enhanced the apparent solubility of both drugs in pure water by 4.7-fold and 5.1-fold, respectively. In vitro dissolution studies indicated an initial rapid release phase, followed by limitations due to gelation. All animal experiments were conducted in strict accordance with the regulations of the Animal Experiment Ethics Committee of Yunnan University of Chinese Medicine, with approval number: YNUTCM-XMSS-G-20250059. Pharmacodynamic evaluation in a zebrafish insomnia model demonstrated that the co-amorphous system at low and medium doses prolonged total sleep time, increased the number of sleep bouts and average bout length, and shortened sleep latency compared to individual drugs or physical mixture. A combination index (CI) < 0.7 confirmed a synergistic sleep-promoting effect. The solvent co-evaporation method proved efficient and straightforward for preparing the co-amorphous system. This co-amorphous strategy demonstrates significant advantages in improving aqueous solubility and achieving synergistic hypnotic efficacy.

Coreopsis tinctoria Nutt. is an important ethnomedicine traditionally used in Xinjiang, China. This study employed ITS2 barcode sequencing for species authentication of commercial C. tinctoria samples. Furthermore, a comprehensive quality evaluation system was established based on HPLC fingerprinting, quantitative analysis of multiple markers, and chemical pattern recognition. A total of 35 batches of C. tinctoria samples were collected from different growing regions in Xinjiang. Genomic DNA was extracted from leaves, and the ITS2 region was amplified by PCR followed by sequencing. The obtained sequences were assembled, proofread, and validated through BLAST analysis against the NCBI database. Subsequently, a neighbor-joining (NJ) phylogenetic tree was constructed based on the aligned sequences. Chromatographic fingerprinting was performed on a ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm). Similarity analysis of the full-spectrum fingerprints was conducted using the "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 version)". Chemometric approaches including cluster analysis (CA), principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA) were employed. Furthermore, a validated HPLC method was developed for simultaneous quantification of four characteristic constituents. The results showed that the main haplotypes of ITS2 sequences in 35 C. tinctoria samples were H1/H2, with a length of 225 bp. The intraspecific variation was small, and the sequences were relatively conservative, indicating that ITS2 sequences can be used as a reliable DNA barcode for the identification of C. tinctoria. The HPLC fingerprinting showed 16 common peaks, and chlorogenic acid, flavanomarein, marein, and isochlorogenic acid A were successfully identified. The similarity of the samples ranged from 0.936 to 0.998. CA classified the samples into three groups, PCA obtained five principal components with a cumulative variance contribution rate of 84.953%, and PLS-DA identified eight components, including chlorogenic acid, flavanomarein, marein, as markers for regional differences. Quantitative analysis showed that the mass fractions of the four components were as follows: Chlorogenic acid 0.860-7.504 mg·g^-1, flavanomarein 3.368-19.182 mg·g^-1, marein 19.719-83.492 mg·g^-1, and isochlorogenic acid A 2.039-12.568 mg·g^-1. The DNA barcoding technique and integrated HPLC fingerprinting/multi-component method demonstrated high reliability and stability. This study provides a scientific foundation for quality control and further research on C. tinctoria.

The rhizosphere soil microenvironment critically influences secondary metabolite accumulation in medicinal rhizomatous plants. To compare the effects of the rhizosphere soil microenvironment in major production areas of Sichuan province on the accumulation of secondary metabolites in turmeric, we collected turmeric from Qianwei, Shuangliu, and Xingwen production areas in Sichuan province as research subjects. UPLC-Q-TOF-MS-based metabolomics was employed to analyze the composition of metabolites. And the high-throughput sequencing platform was utilized to analyze the rhizosphere soil microbial community structure. Then correlation analyses were performed between the metabolites, microbial community, and physicochemical factors. The results indicated that the metabolite profiles of turmeric from different production areas are relatively similar. The most abundant secondary metabolites in plants include ketones, aldehydes, esters, terpenoids, flavonoids, and polyphenols. Polyphenols, terpenoids, and some curcuminoids were significantly higher in turmeric from Shuangliu region than in samples from Qianwei and Xingwen. Significant differences were observed in rhizosphere soil physicochemical properties, including pH, organic matter, total nitrogen, available phosphorus, available potassium, manganese, magnesium, calcium, copper, and selenium among the different regions. At the phylum level, Proteobacteria and Acidobacteria were the dominant bacterial phyla across all three regions, while considerable variations existed at the genus level. In turmeric, correlation analysis revealed that bisdemethoxycurcumin correlated positively with pH, manganese, magnesium, and calcium. ar-Turmerone correlated positively with organic matter and available phosphorus. And tetrahydrocurcumin correlated positively with Rhodanobacter and Sphingomonas. In summary, secondary metabolites differed between turmeric from different regions. These metabolites exhibited correlations with both the physicochemical properties of the rhizosphere soil and the associated microbial communities. These findings provide a scientific basis for cultivation base selection and ecosystem-based cultivation practices.

The heat shock transcription factor (HSF) family plays a crucial role in plant growth, development, and responses to biotic and abiotic stresses. As a traditional Chinese herbal medicine, Glycyrrhiza uralensis Fisch. is rich in various bioactive compounds, including flavonoids and terpenes. However, a systematic identification and characterization of the HSF gene family in G. uralensis is currently lacking. To clarify the biological functions of this gene family, we systematically identified HSF family members based on the genomic data of G. uralensis, and analyzed their physicochemical properties, chromosomal localization, phylogenetic relationships, conserved motifs, and collinearity. Meanwhile, combined with transcriptome sequencing and qRT-PCR analyses, we elucidated the expression patterns of GuHSF genes and their co-expressed genes associated with flavonoid biosynthesis under drought stress. The results showed that a total of 26 GuHSF genes were identified from the genome of G. uralensis, which can be classified into three subfamilies, and the HSF proteins within the C subfamily exhibited high similarity in conserved motifs. Collinearity analysis revealed 12 pairs of segmental duplications within the GuHSF family, and the syntenic relationships between GuHSF genes and soybean HSF genes were the strongest among the compared species. The results of qRT-PCR analysis indicated that GuHSF2, GuHSF4, GuHSF14, GuHSF18, GuHSF21, and GuHSF26 genes were significantly induced by drought stress. In addition, under drought conditions, strong correlations were observed between GuHSF1, GuHSF4, GuHSF9, GuHSF11, and GuHSF21 genes and flavonoid biosynthesis-related genes. This study systematically identified and analyzed the GuHSF family genes in G. uralensis, which provides important clues for subsequent investigations into the biological functions of core members of this family.