The activating transcription factor (ATF) family, a key branch of the basic leucine zipper (bZIP) transcription factors, is widely involved in various biological processes including cellular stress responses, inflammation, apoptosis, and metabolism. In the pathogenesis and progression of pulmonary fibrosis and inflammatory lung diseases, such as acute lung injury/acute respiratory distress syndrome, chronic obstructive pulmonary disease, asthma, ATF family members regulate critical mechanisms including cellular stress, inflammatory responses, and ferroptosis, thereby influencing immune responses and tissue remodeling. They exhibit bidirectional regulatory roles with both pathological promotion and protective effects. This review summarizes the expression changes, biological functions, and potential molecular mechanisms of ATF family members in inflammatory lung diseases and pulmonary fibrosis, aiming to offer novel research perspectives for developing mechanism-based targeted therapies for pulmonary diseases.

Ischemic stroke (IS) remains a leading cause of mortality and disability among adults worldwide, with its core pathological mechanism rooted in ischemic-hypoxic neuronal injury triggered by cerebral blood flow interruption. Current clinical treatments primarily rely on intravenous thrombolysis and endovascular thrombectomy. However, these approaches are constrained by strict therapeutic time windows (typically ≤ 4.5 h), bleeding risks in certain patients, and limited neurorestorative effects following reperfusion. Despite active development of neuroprotective agents and anti-inflammatory drugs in recent years, most have failed in clinical translation due to insufficient efficacy or safety concerns, highlighting the complexity of disease mechanisms and limitations of preclinical evaluation systems. Consequently, developing therapeutics that simultaneously address reperfusion protection and neural repair by targeting the pathological cascade of IS has emerged as a critical direction to overcome current therapeutic bottlenecks. This review synthesizes recent advances in domestic and international IS drug development in recent years, focusing on model establishment, efficacy evaluation metrics, and relevant molecular mechanisms to systematically summarize the preclinical pharmacodynamic evaluation framework for IS therapeutics. The findings provide valuable insights for optimizing preclinical evaluation systems, accelerating drug development, and expanding effective clinical treatment strategies for IS.

As first-line drugs for inhibiting gastric acid and treating digestive system diseases, proton pump inhibitors (PPI) have been found in recent studies to have multi-pathway effects on the regulation of metabolic diseases. There are both therapeutic potentials and risks in the prevention and treatment of metabolic diseases. Studies have indicated that PPI participate in the pathophysiological processes of obesity, type 2 diabetes mellitus (T2DM), and non-alcoholic fatty liver disease (NAFLD) through multiple regulatory mechanisms, including reducing levels of appetite-related hormones, improving insulin sensitivity, decreasing hepatic fat accumulation, and modulating fibrotic progression. However, long-term use of PPI has potential risks, including electrolyte disorders, intestinal flora imbalance, osteoporosis, increased risk of fractures, and increased risk of cardiovascular diseases. In clinical application, the medication regimen should be precisely formulated based on the individual characteristics of patients, long-term use without indications should be avoided, and relevant risks should be closely monitored.

High-throughput screening (HTS) has been widely used in drug discovery recently. It allows batch bioactivity testing of compounds, thereby accelerating the identification of bioactive molecules. However, its screening efficiency and accuracy remain limited. Therefore, artificial intelligence (AI) is gradually integrated into the process of HTS, such as machine learning and deep learning algorithms to enhance ligand-based virtual screening, drug combination screening, and analysis of images, to promote the intelligent development of drug discovery. This review summarized the recent advances in the application of AI technologies in HTS, discussed the applications of machine learning models, and highlighted key AI applications in compound screening and recognition of image in HTS to accelerate the development of innovative drugs.

Tetramic acid unit occurred in a large number of natural products, which normally exhibited antitumor, anti-ulcer, antibacterial, antiviral, and herbicidal activities. Their complex and diverse molecular structures as well as broad biological activities have attracted great attention from biochemists. This article reviewed the natural products with tetramic acid structures isolated since 2013, classified them based on their structures, and introduced their biological activities.

Chemotherapy-induced nausea and vomiting (CINV) represents one of the most prevalent adverse reactions during cancer treatment. Its pathological mechanism is complex, involving multiple neurotransmitters and their biopathways, such as serotonin, substance P, and dopamine, along with their respective receptors. Existing chemical agents for CINV prophylaxis and management face challenges including limited efficacy, serious side effects, and high cost. In contrast, traditional Chinese medicine (TCM) holds considerable promise for CINV prevention and treatment, leveraging advantages such as multi-components, multi-targets, high safety, low side effects, and cost-effectiveness. The structural classes, mechanisms of action, structure-effect relationships, and clinical applications for currently available CINV drugs were systematically reviewed. Moreover, the recent research progress on TCM for preventing and treating CINV was also introduced It will provide significant references for the development of novel CINV therapeutics with sustained efficacy, high efficiency, and low cost.

The voltage-gated sodium channel isoform Nav1.8 is predominantly distributed in peripheral injurious sensory neurons and plays a key regulatory role in the development and maintenance of acute and chronic pain and neuropathic pain. In January 2025, suzetrigine, the first analgesic drug targeting Nav1.8, was approved for marketing in the United States FDA, marking a major breakthrough in the development of Nav1.8-targeted pain therapeutics. The successful development of the first analgesic for Nav1.8 has led to a global interest in the development of Nav1.8 analgesic inhibitors. This article reviews the structure, physiological function, and drug development of Nav1.8. We focus on summarizing the electrophysiological properties of Nav1.8, its role in pain, and the clinical studies of selective inhibitors, aiming to provide a reference for the development of Nav1.8 analgesics.

This article conducted a comparative analysis of the background and fundamental differences between the 505(b)(2) drug and modified new drug, with a focus on successful case studies of 505(b)(2) drugs, discussing clinical requirements and development trends. Based on the national conditions of our country, the project initiation considerations and examples of improved new drugs were carried out from the perspective of preparations. Additionally, the causes of improvement failures and relevant cases were analyzed and discussed, providing critical insights to advance the development of modified new drug in China.

Mitochondria plays an important regulatory role in the anticancer immune response by directly participating in the activation, proliferation and differentiation of immune cells, regulating multiple immune signaling pathways and affecting the activation and function of tumor-infiltrating immune cells with the help of the change of mitochondrial dynamics and metabolic. Mitochondria-targeted immunotherapy is an emerging cancer treatment strategy. The constructed mitochondrial targeted nanoformulation significantly improves the anti-cancer efficacy by inducing immunogenic cell death, regulating mitochondrial dynamics and metabolism, activating immune signaling pathways and other immune strategies. This review focuses on the field of cancer immunotherapy, introduces the relationship between mitochondria and tumor immunity, and summarizes the design of mitochondria-targeted nanomaterials in the past five years. Finally, the challenges and prospects of mitochondria-targeted nanoformulations in anti-cancer immunotherapy are discussed in this paper, which offers support for the development of novel mitochondria-targeted agents for cancer immunotherapy in the future.

It has become an industry consensus to take the concept of quality by design (QbD) as the basic principle of drug development design. On the basis of reviewing the design steps of pharmaceutical QbD, this paper deeply discusses the application of QbD in the development of lipid-based cubic liquid crystalline, and analyzes in detail the QbD elements in the design of lipid-based cubic liquid crystalline, such as quality target product profile, critical material attributes, risk assessment and factor screening, critical material attributes, critical process parameters, experimental design, design space and control strategy, etc. The purpose of this study is to deeply explore the integration and application of QbD concepts and provides a reference for the development of lipid-based cubic liquid crystalline preparations.

Dry granulation is a highly mechanized method for the preparation of drug granules. Roller compaction is a commonly used dry granulation method. The preparation of roller-compacted ribbons is a key step in dry granulation, and the properties of ribbons, such as density, solid fraction, and porosity, is related to the formation rate of drug granules, and directly determine their quality. The conventional method for measuring ribbons properties, which is off-line and time-consuming, can no longer meet the needs of continuous production. The emerging process analysis technology is capable of online real-time detection of ribbons properties. Combined with data analysis technology, it can provide feedback, control, and improve the ribbons preparation process to obtain high-quality ribbons, which is beneficial for continuous production of roller compaction. The method of detecting ribbon properties based on process analysis technology has made significant progress, and has become a hot topic in the analysis and control of roller compaction processes. This article systematically summarizes the latest progress in conventional determination methods and process analysis methods of ribbon properties, in order to provide theoretical guidance for the monitoring of ribbon properties and the continuous production of roller compaction dry granulation.

The intestinal mucus layer is mainly composed of mucin 2 (MUC2), a highly glycosylated protein secreted by goblet cells. The Muc2^‒/‒ mice are a spontaneous colitis model, due to intestinal mucus barrier defects, gut microbes directly contact with intestinal epithelial cells and induce intestinal inflammation. Nevertheless, how gut microbes affect the intestinal epithelium and trigger inflammation in the absence of mucus remains unknown. This study analyzed histopathological changes, colonic transcriptome and gut microbiome of Muc2^‒/‒ mice to elucidate the role of gut microbes in inducing intestinal inflammation. Animal experimentations were approved by the Institutional Animal Care and Use Committee of Nanjing University of Chinese Medicine, Ethical Review No. 202302A007. Pathological analysis showed that Muc2^‒/‒ mice manifested considerable intestinal mucosal damage accompanied by inflammatory cell infiltration and goblet cell exhaustion. Transcriptomic analysis showed that mucus synthesis, endoplasmic reticulum stress, and autophagy-related gene expressions were altered in Muc2^‒/‒ mice. Kyoto Encyclopedia of Genes and Gene Ontology pathway enrichment analyses showed considerable enrichment of inflammation and immune-related pathways, and pathways in response to bacteria derived molecules were upregulated. Microbial gene sequencing revealed dysbiosis of the gut microbiome in Muc2^‒/‒ mice, with Akkermansia muciniphila expansion being the characteristic profile. The expansion of Akkermansia muciniphila was confirmed in Il10^‒/‒ mice, which also displayed mucus barrier defects. Correlation analysis showed that the disturbed gut microbes of Muc2^‒/‒ mice was positively correlated with intestinal inflammation and immune-related differentially expressed genes, which were enriched in pathways such as MAPK cascade and chemokine production. In conclusion, in the absence of intestinal mucus layer, gut microbes can induce immune response by affecting the colonic gene transcription, leading to sustained secretion of inflammatory cytokines and intestinal inflammation. This study investigated the mechanism by which gut microbiota induces colonic inflammation in the absence of the intestinal mucus layer through integrated analysis of colonic transcriptomic profiles and gut microbiota dynamics. The findings aim to identify therapeutic targets for colitis treatment and drug discovery.

As the "power factory" of cells, mitochondria are important organelles for maintaining cellular homeostasis. The depletion of glucose and oxygen causes mitochondrial damage when ischemic stroke occurs, leading to energy metabolism disorders, reactive oxygen species (ROS) aggregation, which further aggravates ischemic nerve injury. Therefore, the aim of this study is to establish a rat middle cerebral artery occlusion/reperfusion (MCAO/R) model in vivo to investigate whether LIG can reduce ischemic nerve injury by restoring energy metabolism and the mechanism. The animal experiment protocol was approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine before implementation (animal ethics number: AHUCM-rats-2021030). TTC staining, neurological function score and brain index were used to evaluate nerve injury in MCAO/R rats. The levels of ROS, mitochondrial membrane potential, ATP, lactic acid and mitochondrial respiratory chain enzyme complex Ⅰ-Ⅳ were detected. The expression of glucose transporter 4 (GLUT 4) was detected by Western blot. Ligustilide was used for molecular docking with AMPK. Western blot was used to detect the protein expression levels of adenosine 5'-monophosphate (AMP)-activated protein kinase [AMPK], p-AMPK, and peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) in HT22 cells damaged by oxygen-glucose deprivation/reoxygenation (OGD/R). The results showed that LIG treatment significantly reduced cerebral infarct volume, brain index and neurobehavioral score in MCAO/R rats. It inhibited the aggregation of ROS, increased mitochondrial membrane potential and ATP content, and reduced lactic acid production. Molecular docking showed that LIG had a good binding ability with AMPK. In addition, LIG increased the protein expression of p-AMPK, AMPK and PGC-1α in HT22 cells damaged by OGD/R. These results suggest that LIG alleviates ischemic brain injury by improving mitochondrial function and restoring energy metabolism, and the mechanism may be related to the activation of AMPK/PGC-1α signaling pathway.

Drug targeting of the human ether-à-go-go-related gene (hERG)-encoded rapid delayed rectifier potassium channel induces acquired long QT syndrome (acLQTS), predisposing individuals to life-threatening arrhythmias. Previous studies demonstrate that the antipsychotic agent thioridazine (THIO) triggers acLQTS. Our findings aimed to elucidate the underlying mechanisms of the therapeutic effects against drug-induced cardiotoxicity. Using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) integrated with multi-electrode arrays (MEA) and optical mapping technology, we evaluated and compared the efficacy of tanshinone IIA (TANIIA), propranolol (Prop) and resveratrol (Res) in reversing THIO-induced prolongation of action potential duration (APD) and field potential duration (all animal experiments were conducted in accordance with the Animal Care Guidelines of Harbin Medical University and were approved by the Ethics Committee of Harbin Medical University, the approval number is IRB3020622). The results showed that TANIIA significantly reversed the THIO-induced prolongation of APD₉₀, APD₅₀, and field potential, whereas neither Prop nor Res counteracted this effect. Building on these results, we further explored the mechanistic basis and molecular targets underlying TANIIA's reversal effects. Western blot and whole-cell patch-clamp demonstrated that TANIIA restored hERG protein expression downregulated by THIO and rescued its channel trafficking defects and rescued channel trafficking defects. Mechanistically, TANIIA attenuated THIO-induced aberrant interactions between hERG and the calnexin, thereby rectifying impaired channel folding and trafficking. Concurrently, TANIIA substantially reduced THIO-induced L-type calcium current. In conclusion, TANIIA ameliorates THIO-induced acLQTS by up regulating hERG current expression and down regulating L-type calcium current. These findings highlight its therapeutic potential of drug-induced acLQTS for clinical.

Ribozymes and DNAzymes are limited in their applications as nucleic acid therapeutics due to their strong dependence on metal ions. Therefore, the development of novel metal-independent ribozymes has become a key research focus. This study evaluated the fidelity of wild-type Tgo DNA polymerase in simultaneously incorporating sugar backbone-modified 2'-deoxy-2'-fluoroarabinonucleic acid (FANA) along with base-modified DNA, constructed a modified nucleotide library containing four modified nucleotides, and employed systematic evolution of ligands by exponential enrichment (SELEX) technology to select ribozymes with RNA cleavage activity. Unlike conventional methods for selecting modified functional nucleic acids, the modified library here was directly used as templates for PCR amplification, eliminating the reverse transcription step which copies the modified phenotypic functional sequence into unmodified cDNA, and dramatically simplified the screening process. This study employed an RNA substrate region composed of 15 consecutive ribonucleosides for selection. Through sequence homology analysis and secondary structure prediction, we verified the cleavage activity of the top twenty abundant sequences obtained from sequencing the most active round 4 library. A sequence denoted R4-11 with considerably high RNA cleavage activity was successfully identified, and it preferentially cleaves the phosphodiester linkage between the rC-rA dinucleotide. This study provides a method for selecting ribozymes composed of diverse modified nucleotides and offers novel tools for disease-targeted therapy through mRNA silencing.

Candida albicans (C. albicans) is the most common opportunistic pathogen in clinical fungal infections, and its invasive pathogenic mechanisms and drug resistance are closely associated with biofilm formation. Therefore, research on biofilm inhibitors holds significant importance. This study found that N-(2,4-dimethylphenyl) maleimide (abbreviated as IMB-D7) can inhibit the formation of C. albicans biofilms and has a certain clearance effect on mature biofilms. IMB-D7 significantly reduces the adhesion capacity and hydrophobicity of C. albicans cells, inhibits the yeast-hyphae phase transition and hyphae formation, and affects the content of key components of the cell wall. IMB-D7 significantly downregulates the expression of genes associated with adhesion, hyphal formation, and cell wall component synthesis. Additionally, IMB-D7 exhibits low cytotoxicity toward human oral epithelial cells. Therefore, further research on IMB-D7 holds promise for the discovery of novel antifungal active lead compounds.

This study systematically investigated the therapeutic mechanism of matrine derivative, (6aS, 10S, 11aR, 11bR, 11cS)-10-methylaminododecahydro-3a, 7a-diaza-benzo (de) anthracene-8-thione (MASM), in experimental autoimmune encephalomyelitis (EAE) through multi-dimensional analysis. The myelin oligodendrocyte glycoprotein MOG_35-55/pertussis toxin-induced EAE mouse model was established, and neurological impairment was confirmed using Kono's 5-point scoring system, mice with scores between 0.5 and 3.5 were considered successful models. These mice were randomly divided into six groups: EAE control, MASM groups (1.5, 3 and 6 mg·kg^-1), a positive control group (fingolimod, 1.0 mg·kg^-1), and a normal control group. Drug administration was initiated on the day of grouping, once daily for 21 consecutive days. The Kono's score was used to evaluate the neurological function; HE staining and electron microscopy was used to observe the pathological changes of spinal cord; transforming growth factor-β1 and monocyte chemoattractant protein-1 content was detected by ELISA. The gene expression of H2T23 was detected by qRT-PCR, the binding interactions between MASM and human frizzled homolog (FZD5) was evaluated by molecular docking, and protein expression levels of glial fibrillary acidic protein (GFAP), complement 3 (C3), wingless type mouse mammary tumor virus integration site family member 5a (Wnt5a), FZD5, protein kinase Cα (PKCα), and the phosphorylation level of glycogen synthase kinase 3 (GSK3β) were detected by Western blot. MASM significantly improved neurological function, reduced spinal cord inflammation and demyelination, and downregulated astrocyte activation markers (GFAP, C3, H2T23) (P < 0.05, P < 0.01) and Wnt5a/FZD5 pathway components (Wnt5a, FZD5, PKCα, p-GSK3β) (P < 0.05, P < 0.01). MASM demonstrates strong binding capacity to FZD. These findings suggest that MASM alleviates EAE by inhibiting astrocyte polarization via the Wnt5a/FZD5 pathway. The animal study was approved by the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine (approval number: FJTCMIACUC 2023319).

M1 macrophages play a critical role in tumorigenesis and development. Insufficient infiltration of M1 macrophages can lead to inadequate CD8⁺ T cell activation, impairing immune surveillance and thereby promoting immune escape. This study investigated the mechanism by which baicalin induces the polarization of RAW264.7 cells towards the M1 phenotype using an in vitro cell model. Cell viability was assessed by the CCK-8 assay. The expression of the M1 surface marker CD86 was analyzed by flow cytometry and immunofluorescence. Cytokine tumor necrosis factor-alpha (TNF-α) levels were measured by ELISA. Lactate levels were detected using a lactate assay kit. Western blot was used to determine the protein expression of key components of the PI3K/Akt/HIF-1α glycolytic pathway and the key rate-limiting glycolytic enzymes lactate dehydrogenase A (LDHA) and pyruvate kinase M2 (PKM2) in RAW264.7 cells. The PI3K inhibitor LY294002 and the glycolytic inhibitor 2-DG were employed to validate the mechanism of action of baicalin. The results showed that baicalin at concentrations of 12.5-100 μmol·L^-1 had no significant effect on the viability of RAW264.7 cells. At 100 μmol·L^-1, baicalin significantly up-regulated the expression of the M1 surface marker CD86 and the cytokine TNF-α, increased the protein expression of p-PI3K, p-Akt, HIF-1α, LDHA, and PKM2, and elevated lactate levels. However, the up-regulatory effects of baicalin on CD86 were inhibited, and the expression of related pathway proteins was reversed, upon addition of LY294002 or 2-DG. This study reveals for the first time that baicalin drives macrophage polarization towards the M1 phenotype by activating the PI3K/Akt/HIF-1α signaling axis and enhancing glycolytic metabolism. This finding provides new mechanistic evidence based on metabolism for its therapeutic potential in tumor immune regulation and inflammation-related diseases.

The phytochemical investigation of the ethyl acetate fraction of 95% ethanol extract of Dendrobium nobile led to the isolation of two previously unreported compounds by various column chromatography methods, including silica gel, MCI, Toyopearl HW-40C combined with the semi-preparative HPLC. Their structures were determined by HRESIMS, NMR, IR, and UV, and identified as neodendronbibisline A (1), neodendronside A (2). Furthermore, compound 1 is a new phenanthrene with a benzofuran structural unit, and compound 2 is a new phenylpropanin glycoside.

The chemical constituents of the soft coral Sinularia sp. collected off Ximao Island in the South China Sea were isolated by comprehensively utilizing various separation techniques such as thin-layer chromatography, silica gel column chromatography, octadecylsilane reverse phase column chromatography, Sephadex LH-20 column chromatography, and high-performance liquid chromatography (HPLC). The planar structure and stereochemistry of the isolated compounds were determined by nuclear magnetic resonance (NMR), mass spectrometry (MS), infrared spectroscopy (IR), and quantum chemical calculations. Six compounds were isolated from the titled soft coral and identified as (3R, 6S)-3-((R)-sec-butyl)-6-isopropylmorpholine-2, 5-dione (1), oculatolide (2), octahydro-4-hydroxy-3R-methyl-7-methylene-R-(1-methylethyl)-1H-indene-1-methanol (3), ent-4(15)-eudesmene-1β, 6α-diol (4), 5-epi-eudesm-4(15)-ene-1β, 6β-diol (5), and ergosterol peroxide (6), respectively. Among them, compound 1 was characterized as a new morpholine derivative, and compounds 3-5 were isolated for the first time from this genus of soft corals. In terms of biological activity, compounds 1-6 underwent screening for antibacterial, anti-inflammatory, and anti-cancer activities, compounds 2 and 6 exhibited significant and broad-spectrum antimicrobial activity, while compound 2 also showed potent anti-inflammatory (IC₅₀ = 5 ± 0.03 mmol·L^-1) and strong anti-proliferative activity against human non-small cell lung cancer cells (A549) and human colorectal adenocarcinoma cells (HT-29) with IC₅₀ values of 17.27 ± 2.32 and 14.14 ± 0.72 mmol·L^-1, respectively.

Employing modern chromatographic techniques, including silica gel, gel chromatography, and semi-preparative liquid chromatography, the chemical constituents from the aerial parts of Salvia hispanica L. were isolated and purified. The structures of the obtained compounds were elucidated by means of contemporary spectroscopic methods. Three neo-clerodane diterpenoids were isolated and identified as 10-deoxyhispanin J (1), ent-(5R, 9R)-15, 16-epoxy-10S-hydroxycleroda-3, 13(16), 14-triene-17, 12S; 18, 19-diolide (2), and dehydrokerlin (3). Among them, compound 1 was a new diterpenoid, and compounds 2 and 3 were isolated from this plant for the first time. The lipid-lowering activity of compound 1 was evaluated in vitro using an oleic acid-induced high-lipid model in HL7702 cells. The results demonstrated that, compared with the positive control curcumin, compound 1 inhibited intracellular lipid accumulation and reduced triglyceride levels, indicating its potential as a lipid-lowering agent.

Three limonoids were isolated and purified from the 95%, 85% ethanol extracts of Melia azedarach using silica gel column chromatography, Sephadex LH-20 column chromatography, MCI column chromatography, and preparative RP-HPLC. Their structures were determined based on physicochemical properties and spectral data, and they were identified as 29-O-ethyl neoazedarachin D (1), 29-O-meliarachin G (2), toosendanin (3). Compound 1 is a new limonoid, while compound 2 is a new natural product isolated from plants for the first time. The in vitro anti-hepatocarcinoma activities of compounds 1-3 were evaluated by MTT assay. The results showed that compounds 1-3 exhibited different degrees of inhibitory effects on both HepG-2 and Hep3B cell lines, and the inhibitory effect of compound 3 on HepG-2 cells was comparable to that of the positive drug cisplatin. In addition, compounds 1-3 exhibited IC₅₀ values of 38.67 ± 1.336, 28.21 ± 1.028 and 8.21 ± 1.231 μg·mL^-1 against HepG-2 cells, and 47.36 ± 1.008, 35.69 ± 1.217 and 15.10 ± 1.017 μg·mL^-1 against Hep3B cells, respectively.

The study aimed to establish a HPLC method and a two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry (2D-LC-Q-TOF/MS) method for the determination of the related substances degradated by tryptophan in pediatric compound amino acid injection and the exploration of the degradation pathway of tryptophan. The HPLC method was carried out on a Waters Atlantis T3 C18 (250 mm × 4.6 mm, 5 μm) column by gradient elution using phosphate buffer solution-acetonitrile (990∶12) and phosphate buffer solution-acetonitrile (65∶35) as the mobile phases to separate tryptophan and the related substances degradated by tryptophan. The selected components were then trapped in switch valve tube lines respectively and delivered to the second-dimensional desalting and deacidification gradient elution which was performed with an Agilent ZORBAX Eclipse Plus C18 (50 mm × 3.0 mm, 1.8 μm) column using acetonitrile-water (95∶5) as the mobile phases. The positive ion mode of the electrospray ionization source was adopted, the parent ions were detected using MS scan mode and the fragment ions were detected using targeted MS/MS mode. The structure of the unknown impurities was figured out by mass spectrometry elucidation. The results showed that tryptophan and its related substances in pediatric compound amino acid injection can be adequately separated under the established analytical conditions. A new impurity produced by the interaction between tryptophan and antioxidant excipients such as sodium bisulfite or sodium metabisulfite was discovered for the first time. The method can provide reference for the quality control of this injection.

Deer antler, a valued traditional Chinese medicine, faces persistent challenges in quality control due to the complex species origins, prevalent adulteration practices, and the absence of specific quantitative standards. To address this challenge, a proteomics strategy was employed to screen signature peptides from deer antlers and reindeer antlers in our pevious study. Combining the reported antler peptides from Cervidae, a comprehensive specificity revalidation was conducted. Finally, six deer antler signature peptides were identified as quality control (QC) markers for quantitative analysis, while reindeer, cow, sheep, and pig-specific peptides were determined as markers for adulteration inspection. Based on these QC markers, an ultrasonic-assisted enzymatic digestion method that completed sample preparation within 6 hours was developed in this study, significantly enhancing analytical efficiency. A UHPLC-MS/MS was utilized for establishment of absolute quantification and adulteration inspection method for deer antlers. The results indictated the validated methods showed high sensitivity and specificity, meeting all requirements for methodological requirements. The proposed method was subsequently applied to 56 commercially available samples (including 28 batches of deer antler slices and 28 batches of deer antler powder). The results revealed that 17.9% for slices and 10.7% for powder were deemed authentic and unadulterated. The total detection rate for the ingredients of reindeer was 73.2% and some samples were found to contain cattle-derived ingredient, indicating the rampant chaos in the deer antler market. This study provided a comprehensive and reliable technical approach to address the aforementioned challenges and achieve quality control for deer antler and its related preparations. The summarized strategy for screening species-spcific peptides and developing quality control methods would provide significant references on the study of quality standards for animal-derived traditional Chinese medicines.

During the development and analysis of low-concentration epinephrine injections, two unreported trace impurities (impurity H and impurity M) have been newly identified. Their analysis and identification were particularly difficult due to their low content, poor stability, and difficult separation in some cases. For impurity H with short retentiontime, poor separation, low content and poor stability, a two-dimensional liquid chromatography-tandem mass spectrometry (2DLC-MS/MS) with column switching technique was employed. This enabled on-line separation, desalination and structure characterization. In contrast, impurity M which exhibits better separability and stability, was isolated and prepared through enrichment and concentration processes. Its structure was then identified using techniques such as liquid chromatography-high resolution mass spectrometry (LC-HRMS) and NMR, enabling the acquisition of more comprehensive structural information. Experimental results showed that impurity H achieved effective separation in the first-dimensional chromatography and successful on-line desalination and fraction cutting in the second-dimensional chromatography. Its potential chemical structure was deduced through multi-stage mass spectrometry data analysis. For impurity M, efficient enrichment and concentration were accomplished; its structure was further confirmed by combining molecular weight and fragment information from LC-HRMS with spectral data from NMR. This study compared the performance of 2DLC-MS/MS with column switching technology with the traditional impurity concentration and identification technology. The experimental results not only provide a valuable strategy for analyzing and identifying trace impurities in complex matrix samples, but also offer an effective analytical tool for the quality control of trace impurities in low concentration epinephrine injections.

Polymer-drug conjugates have shown great potential in the field of drug delivery, but existing systemic pharmacokinetic data are difficult to clarify their pharmacokinetic behavior at the cellular level. In this study, multi-loaded pegylated irinotecan conjugate (PEG-[irinotecan]₃) was used as a model drug to build a research idea based on the interaction of polymorphic components to analyze its cellular pharmacokinetics. An HPLC-MS/MS analysis method based on in-source collision-induced dissociation (IS-CID) technology was established to achieve accurate quantitative analysis of PEG-[irinotecan]₃ and its intracellular polymorphic components, and comprehensively clarify its intracellular uptake, efflux and subcellular distribution characteristics. Uptake studies have shown the presence of polymorphic components in cells such as bonded drugs (PEG-[irinotecan]_{x (x = 3, 2, 1)}), release material (PEG20K), free drug (irinotecan) and active metabolite (SN-38). Subcellular distribution studies have shown that bonded drugs and release materials are mainly distributed in the cytoplasm, while irinotecan is mainly distributed in the cytoplasm and nucleus. This study expands the cellular pharmacokinetic research system of polymer-drug conjugates, and the key information obtained is of great value in guiding drug design, clarifying intracellular pharmacokinetic behavior, and promoting clinical transformation.

Dipeptidyl peptidase 4 (DPP4) is a serine protease widely present in human body, which can specifically catalyze the hydrolysis of proline (Pro) or alanine (Ala) peptide bond at the second N-terminal position of polypeptide chain. It is involved in the activation, partial or complete inactivation of a variety of bioactive polypeptides in vivo. In this study, the kinetic parameters of different probe substrates GP-PNA, GP-AMC and GP-BAN catalyzation by DPP4 were compared, and the inhibitory kinetic differences between DPP4 and its inhibitor sitagliptin/vildagliptin under different probe systems were studied. By means of molecular docking technology, the interaction mechanism between DPP4 and different substrates was revealed from the perspective of small molecules, and the key factors affecting the activity of DPP4 catalytic substrates were discussed. The results show that DPP4 has the best binding property to probe GP-BAN, and its affinity is higher than that of GP-AMC and GP-PNA, which proves that the carbon structure of probe is the key factor affecting the binding. Sitagliptin has better inhibitory activity on DPP4 than vildagliptin. The molecular docking results suggest that the halogenation characteristic of sitagliptin may be the key factor to enhance its binding ability. This study can provide an important basis for the selection of DPP4 probes under different research purposes, and also provide experimental data and research basis for rational design and development of highly selective DPP4 ligand tool molecules and personalized rational drug use in clinic.

In the study, bifunctional tanshinone IIA nanoparticles (Tan IIA/M-NPs) with both mannose targeting and reactive oxygen species response were prepared by nanoprecipitation using methoxypoly (ethylene glycol)-disulfide bond-poly (lactic acid hydroxyacetic acid) copolymer (mPEG_5k-SS-PLGA_60k) and phospholipid poly (ethylene glycol) mannan (DSPE-PEG_5k-Man) as the carriers, and tanshinone IIA (Tan IIA) as the model drug. The preferred process parameters were obtained through one-way experiments, and the optimal conditions were determined as follows: a drug-to-carrier mass ratio of 1∶12, an oil-to-water volume ratio of 1∶10, a peloxam concentration of 2.0%, a sonication time of 12 min, and a targeting material modification ratio of 10%. the average particle size of Tan IIA/M-NPs was 198.3 ± 2.8 nm, and the polydispersity coefficient. The average particle size of Tan IIA/M-NPs was 198.3 ± 2.8 nm, the polydispersity index (PDI) was 0.224 ± 0.002, the zeta potential was -2.62 ± 0.94 mV, and the encapsulation rate was (94.41 ± 0.36)%. The results of in vitro cellular experiments showed that Tan IIA concentration in the range of 0.1-100 µmol·L^-1, Tan IIA, blank vector and Tan IIA/M-NPs were all biocompatible with each other and had no significant toxicity to macrophages; the uptake efficiency of inflammatory macrophages by mannose-modified Tan IIA/M-NPs was significantly higher than that of the unmodified group; and Tan IIA/M-NPs significantly reduced the secretion level of macrophage inflammatory factors (TNF-α, IL-1β), inhibited NO production, decreased the proportion of M1-type macrophages, promoted the polarization of M2-type macrophages, and decreased the intracellular level of reactive oxygen species. Tan IIA/M-NPs had a good drug-carrying capacity, and could enhance the inhibitory effect of Tan IIA via mannose targeting and ROS-responsive release. The inhibitory effect on myocardial inflammation provides a new research idea for the regulation of inflammation and tissue repair after myocardial infarction treated with tanshinone IIA.

Curcumin (CUR), an acidic polyphenolic natural compound extracted from plants of the Zingiberaceae family, exhibits remarkable pharmacological activities such as anti-inflammatory and antioxidant effects. However, its strong hydrophobicity, poor membrane permeability, and low bioavailability severely limit its therapeutic application. In this study, glycyrrhizic acid (GA), a natural compound with surfactant properties, was employed as a carrier to construct curcumin-glycyrrhizic acid self-assembled micelles (CUR-GMS) in order to enhance the solubility and bioavailability of CUR and improve its therapeutic efficacy against ulcerative colitis (UC). Molecular dynamics simulations confirmed that GA and CUR spontaneously formed core-shell structured micelle via hydrophobic interactions and hydrogen bonding, providing theoretical evidence for the feasibility of self-assembly. CUR-GMS were prepared via the thin-film dispersion method and characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), and differential scanning calorimetry (DSC). The micelles exhibited a spherical morphology with a particle size of 46.33 ± 0.63 nm, a zeta potential of 59.76 ± 2.54 mV, an encapsulation efficiency of (98.82 ± 0.41)%, and a drug-loading capacity of (28.24 ± 0.11)%. CUR and GA were present in an amorphous state within the micelles. Compared with the raw drug, CUR solubility was markedly improved; the cumulative in vitro release within 48 h was increased by 2.33 to 3.98-fold, and oral bioavailability was enhanced by 6.36-fold. In vivo pharmacodynamic studies demonstrated that CUR-GMS significantly alleviated symptoms of dextran sulfate sodium (DSS)-induced UC in mice, with the underlying mechanism potentially related to the regulation of macrophage polarization. In conclusion, CUR-GMS constructed with GA significantly improved the oral bioavailability of CUR and enhanced its therapeutic efficacy against UC. All animal experiments were approved by the Ethics Committee of the Animal Experimentation Science and Technology Center, Jiangxi University of Chinese Medicine (No: JZLLSC20230350).

To facilitate the intelligent transformation of traditional Chinese medicine (TCM) manufacturing, this study addresses the challenges in the honey refining process—namely, the reliance on manual experience, difficulties in quantifying multi-dimensional quality attributes, and unclear quality transfer mechanisms. This study, using real-world samples from the production of Tongren Niuhuang Qingxin Pills, established a digital measurement system for physical and chemical quality attributes of the refining process and constructed a pilot-scale quality transfer model. For physical characterization, optimized parameters were determined for dual-mode rheological testing (rotation: shear rate 50 s^-1; oscillation: frequency 5 Hz, strain 50%) to measure honey viscosity. For chemical attributes, a high-performance liquid chromatography (HPLC) method was developed to quantify pH, moisture, fructose, glucose, sucrose, maltose, and 5-hydroxymethylfurfural (5-HMF). Based on 984 data points from both atmospheric and vacuum refining processes, a kinetic model of quality transfer was constructed. The dynamic viscosity (η) showed strong correlations with moisture (S) and temperature (T) (R² > 0.998 3), effectively representing the evolution of physical properties. Additionally, the chemical kinetic model indicated that vacuum refining at low temperature, combined with honey sources low in fructose and monosaccharide/disaccharide ratios, can significantly inhibit Maillard reaction rates. This study presents a systematic digital measurement and modeling framework for the honey refining process, providing technical support for process optimization and quality consistency in TCM manufacturing under the paradigm of intelligent production.

Methicillin-resistant Staphylococcus aureus (MRSA) infection poses a major obstacle to skin wound healing, often accompanied by persistent inflammation and excessive scar formation. There is an urgent clinical need for more effective localized therapeutic strategies. In this study, we developed a responsive nano-oleogel system based on natural bioactive compounds for the synergistic treatment of MRSA-infected wounds. This system consists of self-assembled nanoparticles of hinokitiol and verteporfin (VP), embedded within an oleogel matrix constructed using β-sitosterol. The oleogel conforms well to irregular wound surfaces and enables stimuli-responsive drug release in the presence of reactive oxygen species (ROS) at the infection site. Hinokitiol chelates Fe³⁺ locally, disrupting bacterial iron metabolism and inducing membrane damage and DNA disruption, thereby achieving non-antibiotic bactericidal effects. β-Sitosterol contributes inherent anti-inflammatory activity, helping to modulate early immune responses. VP exerts its effects by inhibiting the Yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) signaling pathway. This multifunctional formulation demonstrates synergistic antibacterial, anti-inflammatory, and scar-modulating effects, offering a promising strategy for the treatment of infected chronic wounds. The animal experiments were approved by the Animal Experimental Research Center of Zhejiang Chinese Medical University and were conducted in accordance with the relevant guidelines and regulations (approval No. IACUC-20241125-07).

With the widespread use of antibiotics, the emergence and spread of drug-resistant bacteria have become a major threat in the global public health field. Small-molecule photodynamic antimicrobial therapy has shown great potential in the treatment of drug-resistant bacteria due to its unique mechanism of action, but small-molecule photosensitizers with near-infrared absorption, which have higher tissue penetration, are still rarely reported. In this study, a cationic small-molecule photosensitizer coumarin-flavylium (CF) with near-infrared absorption was developed, which exhibits excellent anti-drug-resistant bacteria performance. The structure of CF was characterized by ultraviolet-visible absorption spectroscopy, fluorescence emission spectroscopy, etc. The antibacterial effect of CF against methicillin-resistant Staphylococcus aureus (MRSA) was evaluated by the plate coating method, and the antibacterial effect of CF against MRSA was also assessed by experiments such as scanning electron microscopy and LIVE/DEAD^TM staining. The experimental results show that CF has low cytotoxicity, and under the conditions of near-infrared light irradiation (660 nm), low dose (0.5 μmol·L^-1) and low light intensity (20 mW·cm^-2), it exhibits significant antibacterial effect (sterilization efficiency > 99%) and can effectively remove biofilms. This photosensitizer is expected to become a powerful tool to deal with drug-resistant bacterial infections.

This study aims to investigate the compositional characteristics of fresh Panax notoginseng-derived exosome-like nanovesicles (PN-ELNs) and their potential therapeutic effects on psoriasis, providing experimental evidence for plant exosome-based therapeutic strategies to ameliorate psoriasis. PN-ELNs were isolated using differential centrifugation and characterized for morphology and size distribution via transmission electron microscopy and nanoparticle tracking analysis. Their compositional profiles and enriched signaling pathways were analyzed through integrated lipidomics, proteomics, and small RNA sequencing. The in vitro effects on angiogenesis in human umbilical vein endothelial cells (HUVECs) and apoptosis in human keratinocytes (HaCaT) were evaluated. The regulatory effects on lesional symptoms, epidermal hyperplasia, angiogenesis, and immune cell subsets were observed in an imiquimod (IMQ)-induced mouse model of psoriasis. All animal experiments were conducted in accordance with the "Regulations on Laboratory Animals" issued by the National Science and Technology Commission of China and the protocol approved by the Animal Experiment Ethics Committee of Chengdu University of Traditional Chinese Medicine (approval No. 2020DL-126). Results demonstrated the successful isolation of PN-ELNs from fresh Panax notoginseng rhizomes, exhibiting a characteristic bilayer membrane structure with an average diameter of 143.9 nm. Multi-omics analyses identified 30 lipid subclasses, 196 proteins, and 6 056 miRNAs, with enriched pathways such as oxidative phosphorylation and amino acid metabolism being closely associated with psoriasis pathogenesis. PN-ELNs significantly inhibited HUVEC angiogenesis and promoted HaCaT cell apoptosis (P < 0.05). In the psoriatic mouse model, PN-ELNs significantly ameliorated skin lesions [reduced psoriasis area and severity index (PASI) scores], attenuated epidermal thickening, suppressed angiogenesis, and modulated the CD4⁺/CD8⁺ immune balance. In conclusion, PN-ELNs exert anti-psoriatic effects through multi-target mechanisms (inhibiting angiogenesis, promoting keratinocyte apoptosis, and improving the immune microenvironment). This study provides the first systematic elucidation of the compositional characteristics of PN-ELNs and their anti-psoriasis potential, offering theoretical support and experimental evidence for developing novel therapeutic strategies based on plant-derived exosomes.

Natural starch granules exhibit limited stability in Pickering emulsions due to their strong surface hydrophilicity. In this study, debranched Smilax glabra starch was used as a matrix to prepare starch-fatty acid complexes (SGS-LA, SGS-LOA, and SGS-SA) by complexation with lauric acid (LA), linoleic acid (LOA), and stearic acid (SA), respectively. Their physicochemical properties and emulsification performance were systematically evaluated. Scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction analyses indicated that these complexes formed Ⅴ-type crystalline structure through non-covalent interactions. Among them, SGS-LA exhibited the highest complex index (46.07%), with an average particle size of 114.30 μm and a contact angle of 68.71°. In the Pickering emulsions system, SGS-LA demonstrated the best emulsifying performance. Under the conditions of 80% oil phase volume fraction and 2% particle concentration, the emulsion stabilized by SGS-LA had the smallest droplet size (approximately 10 μm) and the highest emulsification index (82.87%). Environmental stability evaluation further revealed that neutral pH and low ionic strength (0-25 mmol·L^-1) were conducive to emulsion stability. This study demonstrates that short-chain fatty acids facilitate the formation of starch-fatty acid complexes with small particle size and good emulsification performance, while also highlighting the important influence of environmental conditions on emulsion stability. These findings provide new insights for developing sustainable natural emulsifiers.