JAZ proteins, as inhibitors in the jasmonic acid (JA) signaling pathway, play a crucial role in regulating plant growth and development, as well as the biosynthesis of secondary metabolites in plants. To explore the functions of the JAZ gene family in Tripterygium wilfordii, our study systematically identified the members of the JAZ gene family in T. wilfordii. We analyzed their physicochemical properties, chromosome localization, phylogenetic tree, and protein conserved motifs. Additionally, we investigated the expression patterns of TwJAZs in different tissues, constructed a gene regulatory network map, and ultimately cloned the full-length sequences of 18 JAZ genes. The results showed that the 18 TwJAZs were unevenly distributed across 12 chromosomes, encoding amino acid numbers ranging from 122 to 525, with molecular weights of 13.87 to 54.73 kDa, and isoelectric points ranging from 6.83 to 10.27. Subcellular localization prediction indicated that 13 TwJAZs were localized in the nucleus, while 5 TwJAZs were located in the cytoplasm and chloroplasts. The phylogenetic tree analysis revealed that the JAZ family proteins of T. wilfordii could be divided into five subfamilies, with members of the same subfamily sharing similar conserved motifs. Expression pattern analysis demonstrated that TwJAZs were predominantly expressed in flowers, leaves, and peeled stems, with most TwJAZs showing the highest expression levels after 4 hours of methyl jasmonate (MeJA) induction. The transcription factor regulatory network shows that TwJAZs are strongly correlated with transcription factor families such as AP2/ERF-ERF, NAC, and bHLH. This study comprehensively identified all the JAZ gene family sequences in T. wilfordii, initially clarifying the structural and functional characteristics of the TwJAZs, and laying an important foundation for further research on the functions and regulatory mechanisms of TwJAZs.

Exosomes are small vesicles secreted by cells that contain important bioactive molecules such as nucleic acids and proteins. Increasing research indicates that exosomes play a unique and crucial role as signal molecule carriers in various diseases, and they exhibit great potential in disease diagnosis and treatment. Recent studies have shown that exosomes play an important role in immune regulation. The NOD (nucleotide binding oligomerization domain)-like receptors protein 3 (NLRP3), an essential component of the innate immune system, plays a key role in the occurrence and development of various diseases including autoimmune diseases, metabolic diseases, and neurodegenerative diseases. Its activation and regulatory mechanisms are complex and diverse. However, the regulatory mechanisms associated with exosomes and NLRP3 inflammasome have not yet been fully elucidated. This article reviews the regulatory effects of exosomes from different sources on the NLRP3 inflammasome and summarizes the therapeutic potential of exosomes in diseases associated with the NLRP3 inflammasome, aiming to provide new ideas for the prevention and treatment of diseases related to the NLRP3 inflammasome.

To establish the cells stably co-expressing NOD1 receptor and enhanced green fluorescent protein (EGFP)-tagged nuclear factor of activated T cells 2 nuclear factor (NFAT2) (EGFP-NFAT2) in U2OS cell, the NOD1 (NM_006092) pcDNA3.1-3×Flag-hygro recombinant plasmid was transfected into U2OS-EGFP-NFAT2 cells, which were screened by pressure of hygromycin B and then incubated with NOD1 agonist lipopolysaccharides (LPS) for 30 min, and the green fluorescence intensity in the nucleus of the cells was detected by the high content screening assay. There were 46 cell strains expressing NOD1 in U2OS-EGFP-NFAT2 cells by EGFP-NFAT2 nuclear translocation assay. Among these cells, cells No 4 had the highest nuclear translocation function. Therefore, it was selected as the U2OS-EGFP-NFAT2-NOD1 cell for functional validation. The expression levels of NOD1 mRNA and protein in the selected U2OS-EGFP-NFAT2-NOD1 cells and the control cell U2OS-EGFP-NFAT2 were examined by real-time quantitative PCR (RT-qPCR) and Western blot. The results showed that NOD1 mRNA was stably expressed in this stably transfected cell line for 5-20 generations, and NOD1 protein was expressed in U2OS-EGFP-NFAT2-NOD1 stably transfected cell line, whereas no NOD1 protein was expressed in the control cell U2OS-EGFP-NFAT2. U2OS-EGFP-NFAT2-NOD1 cells were treated with histones or LPS for 30 min, and the EGFP-NFAT2 nuclear translocation was detected by the high content screening assay. Histones were found to significantly increase the EGFP-NFAT2 nuclear translocation in U2OS-EGFP-NFAT2-NOD1 stably transfected cells over a range of concentrations. The U2OS-EGFP-NFAT2-NOD1 cells were divided into the solvent control group, NOD1 receptor antagonist nodinitib-1+histone group, and histone group. The drug incubation time was 30 min, and the specificity of the NOD1 cells was verified by observing the EGFP-NFAT2 nuclear translocation through the high content screening assay. Compared with the histones group, the nodinitib-1+histones group significantly decreased EGFP-NFAT2 nuclear translocation in U2OSEGFP-NFAT2-NOD1 cells (P < 0.05). In conclusion, U2OS-EGFP-NFAT2-NOD1 cells stably co-expressing NOD1 and EGFP-NFAT2 are established, which can be used for screening antagonistic compounds targeting NOD1 pathogenic microorganisms with mechanism study.

Immune checkpoint inhibitors (ICIs) have emerged as critical agents in cancer immunotherapy; however, their resistance and limited response in most patients pose significant challenge. The gut microbiota, as a pivotal immune regulator, has been increasingly recognized for its role in enhancing the therapeutic efficacy of ICIs. Studies demonstrate that fecal microbiota transplantation or transplantation of specific bacterial strains can directly reshape the gut microbiota composition, thereby improving ICI therapeutic outcomes. Furthermore, dietary interventions, prebiotics, and postbiotics have shown potential in augmenting the anti-tumor effects of ICIs through gut microbiota modulation. Despite these promising findings, further investigations are required to optimize microbiota-based strategies and therapeutic protocols. This review highlights the critical role of gut microbiota modulation in ICI-based cancer therapy and explores its clinical applications, offering both practical insights and theoretical foundations for improving immunotherapy outcomes against various cancers.

To investigate the effect and underlying mechanism of action of Yueju volatile oil (YJVO) in the treatment of high altitude sleep disturbance (HASD) mice, gas chromatography-mass spectrometry (GC-MS) was used to identify the components of YJVO, while network pharmacology was applied to predict the mechanism of action. KM mice were selected and randomly assigned to several groups: the control group (control), the model group (model), and the YJVO treatment groups at low (YJVO-L, 200 μL·m^-3), medium (YJVO-M, 400 μL·m^-3), and high (YJVO-H, 800 μL·m^-3) doses, as well as the diazepam group (DZP, 2 mg·kg^-1). Except for the control group, all mice were subjected to a hypobaric oxygen chamber to establish the HASD model. Animal experiments were approved by the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics No.: TEMPOR20230088). Co-sleeping and weight-bearing swimming experiments were conducted to assess the sleep-regulating effects of YJVO. Hematoxylin and eosin (H&E) staining was employed to observe damage in hypothalamic and hippocampal brain tissues. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure levels of melatonin (MT), γ-aminobutyric acid (GABA), interleukin-6 (IL-6), interleukin-1beta (IL-1β), 5-hydroxytryptamine (5-HT), orexin-A, and tumor necrosis factor-α (TNF-α) in both plasma and brain tissues. Concurrently, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were detected to evaluate oxidative stress. Western blot was used to determine the expression of key proteins in the NF-κB/NLRP3 pathway, and immunohistochemistry (IHC) was used to detect the expression of 5-hydroxytryptamine 1A receptor (5-HT_1A) in hippocampal and hypothalamic tissues. The GC-MS results revealed that YJVO was identified to contain 68 components. Network pharmacology results indicated that the mechanism of YJVO in treating HASD involves multiple signaling pathways such as AGE-RAGE, TNF, serotonin, and NF-κB. Behavioral experiments indicated that YJVO significantly prolonged the sleep duration, reduced the sleep latency, and bolstered the physical endurance and anti-fatigue capabilities of mice (P < 0.01). H&E staining results showed significant improvement in the pathological damage of the hippocampus and hypothalamus tissues in HASD mice. ELISA results indicated that YJVO increased the concentrations of sleep-inducing neurotransmitters MT and GABA within brain tissues, decreased the levels of wakefulness-inducing neurotransmitters orexin-A and 5-HT in plasma, and attenuated the secretion of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α (P < 0.01). Biochemical results indicated that YJVO could inhibit the production of MDA in the brain tissue of HASD mice and enhance the activity of SOD and GSH-PX (P < 0.01). Western blot results showed that YJVO downregulated the protein expression of phosphorylate nuclear factor-kappa B (p-NF-κB p65)/nuclear factor-kappa B (NF-κB p65), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase 1 (Caspase-1), and IL-1β in the brain tissue of HASD mice (P < 0.01). IHC results demonstrated that YJVO downregulated the protein expression of 5-HT_1A in the hippocampus and hypothalamus tissues of HASD mice (P < 0.01). The study findings indicate that YJVO demonstrates potent therapeutic effects in HASD mice, potentially through the modulation of neurotransmitters and the attenuation of neuroinflammation.

Three new compounds were isolated from the n-butanol fraction of Zingiber officinale (Ginger) peel by MCI Gel CHP-20, Sephadex LH-20, ODS and semipreparative high performance liquid chromatography. Their structures were identified as dendranthemoside C (1), zingpyranoside E (2), zingpyranoside F (3) by 1D-NMR, 2D-NMR, HR-ESI-MS, circular dichroism (CD) and ECD calculation techniques.

Through several chromatographic methods, such as silica gel, Sephadex LH-20 and high performance liquid chromatography (HPLC), six iridoids were isolated from the roots and rhizomes of Patrinia scabiosaefolia. By means of nuclear magnetic resonance (NMR) spectroscopy, ¹³C NMR calculation and mass spectrometry (MS), their structures were identified as patrinin A (1), patrinin B (2), loganin aglycone (3), isovillosol (4), 1, 3-dimethoxy-4, 7-dimethyl-octahyhro-cyclopenta[c]pyran-6, 7-diol (5) and viburnshosin A (6). Among them, compounds 1 and 2 were new compounds. Furthermore, the anti-influenza virus and anti-inflammatory activities of the isolates were evaluated.

Through literature analysis, the advantages of oral pharmaceutical vehicles in solving the temporary dispensing problems for special populations are summarized. Detailed introduction is made on the research, development, production, use, and regulatory experience of oral pharmaceutical vehicles in the United States, as well as the current research progress, achievements, and challenges faced by domestic oral pharmaceutical vehicles. The United States has formed a mature system in the field of oral pharmaceutical vehicles, and China can learn from the experience of the United States to improve relevant laws and policies, promote the establishment of technical guidance principles and technical standards for the use of oral pharmaceutical vehicles in temporary dispensing, and accelerate its clinical use.

Based on the genome and transcriptome data of Salvia miltiorrhiza, a cytochrome P450 gene annotated as SmCYP72A395, which was highly expressed in the periderm of S. miltiorrhiza root was cloned. The total length of the cDNA sequence of SmCYP72A395 was 1 578 base pairs, encoding 525 amino acids. The predicted molecular weight of the protein was 59.9 kDa. The physicochemical properties, subcellular localization, protein structure, and conserved domains were predicted by online bioinformatic tools. The theoretical isoelectric point of SmCYP72A395 was 8.68, with a transmembrane domain in the protein. This gene was highly expressed in the flower, leaf, and root periderm tissues of S. miltiorrhiza. To further identify the biological function of SmCYP72A395, the transgenic hairy roots with SmCYP72A395 overexpressed (SmCYP72A395-OE) and RNA interference (SmCYP72A395-RNAi) were constructed, and the content of tanshinone compounds was detected by UPLC between these transgenic lines and control line (haboring the vector plasmid in the transgenic line). Compared with the control line of transgenic hairy roots, the content of dihydrotanshinone Ⅰ, cryptotanshinone, tanshinone Ⅰ and tanshinone ⅡA in the SmCYP72A395-OE lines was significantly less than those in the control line. On the contrary, the content of dihydrotanshinone Ⅰ, cryptotanshinone, and tanshinone Ⅰ was higher in the SmCYP72A395-RNAi lines than those in the control line. These results demonstrated that SmCYP72A395 played a negative role in the regulation of tanshinone accumulation in S. miltiorrhiza. This study lays the foundation for further elucidating the biosynthesis and regulatory pathways of tanshinone compounds in S. miltiorrhiza.

In 2024, drug discovery continues to develop on the basis of the achievement before. The number of approved first-in-class (FIC) drugs reach a higher level. The Center for Drug Evaluation and Research of U.S. Food and Drug Administration has totally approved 50 novel drugs, including 30 small molecule drugs and 20 macromolecule drugs. Comparing to the number of approved drugs in 2023, the total in 2024 falls slightly. But the proportion of FIC drugs increase obviously. There are 24 FIC drugs (48%) approved in 2024, which include 13 FIC small molecule drugs and 11 FIC macromolecule drugs and whose indications mainly focus on tumor, and endocrine and metabolic disease. Among 24 FIC drugs, many drugs have breakthrough clinical value, such as the first combination of muscarinic acetylcholine receptor M1/M4 agonist and peripheral muscarinic acetylcholine receptor antagonist, xanomeline and trospium chloride, the first thyroid hormone receptor beta agonist, resmetirom, the first dual antagonist of endothelin receptor type A and B to treat hypertension, aprocitentan. The huge value of FIC drugs in clinical therapy, academic research and commerce, is attracting wide attention from the patients, researchers and enterprises. This review will analyze the research background, development process and therapeutic application of three first-in-class small molecule drugs in this year from a medicinal chemistry perspective, expecting to provide more research directions and methods for the development and research of FIC drugs.