In 2025, innovation in drug development remained stable, with the proportion of first-in-class drugs continuing to increase. The Center for Drug Evaluation and Research (CDER) of the US Food and Drug Administration (FDA) approved a total of 46 new drugs in the past year, including 34 new molecular entities (NMEs) and 12 biologics license applications (BLAs). The number of approvals slightly decreased compared to the previous three years. This year, 31 small molecule drugs were approved, accounting for 67% of the total, among which 13 were first in class small molecule drugs. The treatment areas covered rare diseases, oncology, autoimmune diseases, respiratory diseases, and more. Several first in class small molecule drugs have breakthrough clinical value, such as the world's first non-opioid analgesic drug, NaV1.8 inhibitor suzetrigine, the first approved local TRPM8 receptor agonist acoltremon for dry eye disease, the first targeted mitochondrial inner membrane cardiolipin elamipretide, the first approved selective phosphodiesterase 4B (PDE4B) inhibitor nerandomilast for idiopathic pulmonary fibrosis (IPF), and the first selective oral Menin-MLL inhibitor iftomenib. First in class drugs have significant clinical, academic, and commercial value and have attracted extensive attention from patients, researchers and enterprises. This article briefly analyzes the research background, process and therapeutic applications of three representative first in class small molecule drugs from the perspective of medicinal chemistry, with the aim of providing more research directions and methods for the development of first in class drugs.

The 2025 edition of the Chinese Pharmacopoeia defines films as film-like preparations made by processing active pharmaceutical ingredients with suitable polymers, intended for oral or mucosal use. Among these, orodispersible films (ODFs) are specifically defined as films that rapidly dissolve in the oral cavity to exert systemic or local effects. ODFs circumvent the drawbacks associated with orally disintegrating tablets, such as fragility and the potential risk of choking. This dosage form has aroused significant interest among both academic world and the pharmaceutical industry due to its accurate dosing, portability, and suitability for elderly and pediatric patients or those with swallowing difficulties. This review summarizes the advantages and limitations of ODFs, their listing status, suitable active pharmaceutical ingredients, commonly used excipients, preparation techniques, and emerging nanoparticle technologies. Additionally, it emphasizes quality evaluation requirements—such as dissolution, disintegration time, and mechanical properties—tailored to the characteristics of this dosage form. The aim is to provide readers and researchers with a comprehensive understanding of the development and evaluation of ODFs.

This study focuses on the innovative applications of microneedles in transdermal drug delivery, aiming to explore their underlying mechanisms for overcoming skin barrier limitations and summarise cutting-edge advances. Recent transdermal delivery research has increasingly emphasised mechanism optimisation and material innovation for microneedles. This review analyses five delivery mechanisms — including "poke and patch" and "coat and poke" — clarifying their principles, advantages, and existing challenges. By examining application cases across ophthalmic disorders, dermatology, oncology, autoimmune diseases, and chronic metabolic conditions, it demonstrates broad prospects for drug delivery efficiency and health monitoring. Future efforts must address critical challenges including scalable production, long-term safety validation, and personalised treatment approaches to accelerate clinical translation.

Bile acids (BAs) have evolved from their traditional role as auxiliary molecules in lipid digestion to critical signaling molecules regulating host metabolism. In recent years, the role of microbiota-derived bile acid-modifying enzymes and their mediated novel bile acid metabolic pathways in the pathogenesis and progression of metabolic diseases has become increasingly prominent. These microbial enzymes fine-tune glucose and lipid metabolism, energy homeostasis, and inflammatory responses by modulating the composition and proportion of the bile acid pool, thereby influencing the activity of key receptors such as the farnesoid X receptor (FXR) and the G protein-coupled bile acid receptor (TGR5). The theory of "harmonizing the liver and spleen" in traditional Chinese medicine (TCM) is closely related to the maintenance of homeostasis of the liver-spleen axis. TCM posits that "liver Qi failing to ensure free flow" (liver dysfunction) and "spleen failing to transport and transform" (spleen dysfunction) are the core pathogenesis of metabolic diseases, while modern research has confirmed their intrinsic pathophysiological links to bile acid metabolic disorders and gut microbiota dysbiosis. Studies have shown that TCM formulations and active ingredients based on the "harmonizing the liver and spleen" principle can effectively ameliorate metabolic disorders such as insulin resistance and lipid accumulation by targeted intervention of microbiota-derived bile acid-modifying enzymes, regulating downstream receptor signaling pathways like FXR and TGR5, consequently correcting bile acid metabolic abnormalities and remodeling the gut microbiota structure. This review systematically summarizes the key targets of the microbiota-derived bile acid-receptor axis in metabolic diseases and delves into the intervention mechanisms of "harmonizing the liver and spleen" TCM strategies based on this axis, aiming to provide a new scientific explanation and potential therapeutic strategies for the prevention and treatment of metabolic diseases with TCM.

Microreactors and related technologies, as key components of continuous manufacturing, offer significant advantages in pharmaceutical production, including enhanced manufacturing flexibility, substantial cost reduction, shortened reaction times, improved production efficiency, environmental sustainability, and strengthened supply chain resilience. They are widely regarded as a core enabling technology for realizing continuous pharmaceutical manufacturing and driving the transformation and upgrading of the pharmaceutical industry. In particular, the integration of microreactors with artificial intelligence and automation technologies has reshaped the traditional large-scale production paradigm of "reactor-production line-manufacturing plant", giving rise to new characteristics such as intelligent controllability, operational flexibility, and mobility, as well as modular and stepwise integration, and on-demand production. This paper systematically reviews the performance characteristics of various types of microreactors in pharmaceutical synthesis and their potential applications in intelligent, continuous pharmaceutical manufacturing. Furthermore, it analyzes the development of intelligent and distributed pharmaceutical synthesis systems enabled by the deep integration of artificial intelligence and chip-based microreactors, aiming to achieve continuous, precise, and emergency-responsive drug production, thereby providing important support for the digitalization and intelligent transformation of pharmaceutical manufacturing.

Diabetic kidney disease (DKD) is a progressive renal complication for which effective reversal strategies remain elusive. Traditional Chinese medicine (TCM), based on the principle of "strengthening the body's defenses and expelling pathogens" demonstrates advantages in multi-targeted, holistic regulation for controlling blood glucose and improving renal function. Astragalus membranaceus, a key Qi-tonifying herb, excels at replenishing Qi, elevating Yang, promoting diuresis, and reducing edema. Its properties align closely with the core pathogenesis of DKD—Qi deficiency with retention of turbidity—making it a frequently used clinical agent for preventing and treating DKD. This systematic review of relevant literature reveals that Astragalus membranaceus, through its active components (e.g., astragaloside Ⅳ, astragalus polysaccharide, and calycosin), synergistic combinations (e.g., Astragalus membranaceus-Dioscorea opposita, Astragalus membranaceus-Cornus officinalis, Astragalus membranaceus-Caragana sinica), and formulae (e.g., Shenqi Dihuang Decoction, Yuye Decoction, Huangqi Gegen Decoction, Huangqi Liuyi Decoction, Huangqi Decoction), and related proprietary Chinese medicines like Keluxin Capsule, Shenqi Jiangtang Capsule, and Niaoduqing Granule demonstrate promising potential in DKD prevention and treatment. Its mechanisms involve regulating glucose and lipid metabolism, inhibiting oxidative stress and inflammatory responses, improving insulin resistance, and modulating multiple pathways of programmed cell death (pyroptosis/apoptosis/ferroptosis/autophagy), counteracting renal fibrosis, and alleviating endoplasmic reticulum stress. This review systematically summarizes research progress on Astragalus membranaceus and its formula combinations in intervening DKD, aiming to preliminarily elucidate the modern scientific implications of Astragalus membranaceus's "Qi-tonifying and water-draining" efficacy, providing reference for its clinical application and further investigation.

Schisandra chinensis, a traditional Chinese materia medica, possesses unique pharmacological activities and extensive clinical application value. Its processing techniques have evolved and improved over thousands of years. In the traditional Chinese medicine system, the processing of Schisandra chinensis directly affects its efficacy and is closely related to quality control and medication safety. Currently, there are still deficiencies in the standardization of processing techniques, quality control of active components, and research on the component-efficacy relationship. This review summarizes the historical evolution and modern technological advancements in the processing of Schisandra chinensis. Integrating contemporary analytical techniques and pharmacological studies, it discusses the quality control methods for its main active components, the mechanisms by which different processing methods influence these components and their efficacy, as well as its applications in metabolic diseases, liver diseases, respiratory diseases, and neurological disorders. This paper aims to provide a reference for the standardization of processing techniques, the precision of quality control, and the modernization of clinical applications of Schisandra chinensis.

Deciphering the "multi-component, multi-target" synergistic regulation of traditional Chinese medicine (TCM) is a pivotal scientific challenge, particularly given ongoing debates over its relevance to holistic therapeutic outcomes and disease prognosis. Building on the framework of Molecular Holistic State Medicine and the concept that mesoscale key functional modules, termed functional assemblies, serve as the structural basis for TCM's holistic efficacy, this study employs "Xiaoke", a condition where TCM demonstrates recognized therapeutic advantages, and the classical formula Xiaoke Prescription as a proof-of-concept model. Guided by the translational principle of "from clinical observation, through experimental validation, back to clinical confirmation", the investigation starts with a retrospective matched clinical cohort to characterize the molecular holistic profile of diabetes and its evolution under Xiaoke Prescription treatment. These clinical findings are then integrated with mechanistic studies to identify the mesoscale key functional modules responsible for Xiaoke Prescription's antidiabetic effects and to reveal the nonlinear regulatory dynamics through which multiple bioactive components modulate multiple targets within these modules. Finally, prospective clinical trials are conducted to verify the synergistic antidiabetic mechanisms of Xiaoke Prescription through the holistic response of the identified mesoscale modules. By shifting mechanistic interpretation from "macroscopic phenotypes" and "microscopic molecules" to "systems-level integration", this study proposes a novel paradigm and technical platform for unraveling the "multi-component, multi-target" synergistic mechanisms underlying TCM interventions.

This study systematically characterized the chemical composition of Yuanhu Zhitong Oral Liquid (YZOL) and elucidated its therapeutic mechanism in primary dysmenorrhea (PD). Using UPLC-Q-TOF-MS/MS, a total of 100 constituents were preliminarily identified in YZOL, predominantly isoquinoline alkaloids and coumarins. A rat model of PD was established by combined administration of estradiol benzoate and oxytocin, and the pharmacological effects of YZOL were comprehensively evaluated through behavioral assessment, quantification of dysmenorrhea-related mediators, and histopathological examination. The results demonstrated that YZOL significantly reduced writhing frequency and prolonged writhing latency in PD rats (P < 0.05). Specifically, medium- and high-dose YZOL markedly decreased the PGF_2α/PGE₂ ratio (P < 0.05), while all dose groups significantly reduced the TXB₂/6-keto-PGF_1α ratio (P < 0.05). Consistently, YZOL treatment markedly ameliorated uterine histopathological damage, with the high-dose group exhibiting tissue morphology closest to normal. Transcriptomic profiling of uterine tissues identified 403 genes reversed by YZOL treatment, including 280 upregulated and 123 downregulated genes in the YZOL-H versus PD comparison. KEGG pathway enrichment analysis indicated that these reversed genes were primarily involved in the TGF-β, Rap1, PI3K-Akt, and MAPK signaling pathways, as well as ovarian steroidogenesis. Functional annotation further revealed that these pathways are closely associated with nociceptive signaling, inflammatory responses, synaptic activity, and hormone biosynthesis, and that their aberrant activation contributes to pain sensitization, inflammatory mediator release, and excessive uterine smooth muscle contraction during PD progression. Integrative analysis combining transcriptomics and network pharmacology identified seven core targets associated with key pathological processes, including pain transmission, inflammatory regulation, and uterine smooth muscle contraction. Molecular docking analysis using AutoDock Vina demonstrated that berberine, peucedanoside, bakuchiol alcohol, and other constituents exhibited strong binding affinities toward these targets, with binding energies comparable to or exceeding those of positive control compounds. In addition, several compounds showed favorable oral bioavailability, suggesting their potential contribution to the in vivo efficacy of YZOL. Based on bioinformatics analysis and batch molecular docking, a multidimensional "active component-key target-biological effect" network was constructed to systematically illustrate the therapeutic mechanism of YZOL against PD. Collectively, these findings indicate that YZOL exerts anti-dysmenorrheic effects through a multi-component, multi-target, and multi-pathway mode of action, in which key constituents including berberine, peucedanoside, bakuchiol alcohol, dihydrochelerythrine, and imperatorin may act on targets such as IL1B, PRKCG, TRPV1, and PTGER2 to coordinately modulate pain signaling, inflammatory responses, and uterine smooth muscle contraction. The experiment was approved by the Laboratory Animal Ethics Committee of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (approval No. 2021B020).

To evaluate the antitumor activity of the novel antibody-drug conjugate (ADC) hIMB1636-MMAE against human epidermal carcinoma, this study systematically investigated its therapeutic efficacy and mechanism of action in A431 cells. Trophoblast cell surface antigen 2 (TROP2) expression in A431 cells was confirmed by Western blot. Flow cytometry was used to analyze the binding characteristics of hIMB1636-MMAE to A431 cells and its effects on cell cycle arrest and apoptosis. The internalization process of the ADC was visualized using confocal laser scanning microscopy, while cytotoxicity in vitro was assessed via the CCK-8 assay. Moreover, an A431 cell-derived xenograft model was established in nude mice to evaluate the in vivo antitumor efficacy of hIMB1636-MMAE. Results showed that A431 cells exhibited high TROP2 expression. hIMB1636-MMAE could specifically bind to A431 cells, and was effectively internalized and transported to lysosomes. Furthermore, hIMB1636-MMAE specifically killed cancer cells with IC₅₀ value of 1.49 ± 0.22 nmol·L^-1, and induced G2/M phase cell cycle arrest and apoptosis. In A431 cell-derived xenografts, hIMB1636-MMAE at 10 mg·kg^-1 exhibited a significant antitumor effect, achieving a tumor inhibition rate of 93.89% (all animal protocols were approved by the Animal Care and Use Committee at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, No. IMB-20220210D₂03). In conclusion, hIMB1636-MMAE demonstrates promising potential against skin cancer and may serve as a candidate therapeutic for TROP2-positive cutaneous malignancies.

This study aimed to investigate the hepatoprotective effects of the combination of berberine (BBR) and stachyose (Sta) in spontaneous type 2 diabetic KKAy mice. KKAy mice were randomly divided into four groups (n = 14 per group): Model control (model), stachyose (Sta, 200 mg·kg^-1), berberine (BBR, 100 mg·kg^-1), and BBR + Sta (100 mg·kg^-1 BBR + 200 mg·kg^-1 Sta), with oral gavage of water, Sta, BBR, and BBR + Sta respectively for eight weeks. At the end of the experiment, serum and liver tissues were collected, the liver weight was measured, and liver index was calculated. Lipid content and oxidative stress-related indicators in liver were measured. Meanwhile, the levels of lipid, transaminase, and oxidative stress in serum were determined. The expression levels of genes related to lipid metabolism and inflammation in the hepatic tissues were detected using qPCR. The number of macrophages in the liver tissues was measured by flow cytometry. Our findings revealed that the combination of BBR and Sta significantly reduced the liver index, decreased hepatic contents of triglyceride (TG), total cholesterol (TC), nitric oxide (NO), inducible nitric oxide synthase (iNOS), and malonaldehyde (MDA), improved liver histomorphology, attenuated lipid deposition, lowered serum lipids and alanine aminotransferase (ALT) levels, and suppressed the expression of lipid synthesis-related genes [fatty acid synthetase (Fasn), sterol regulatory element-binding protein 1c (Srebp1c)] and inflammation-related genes [interleukin-6 (Il-6), tumor necrosis factor-α (Tnf-α), monocyte chemoattractant protein-1 (Mcp-1)] in hepatic tissues. Furthermore, the combination treatment reduced the total number of hepatic macrophages (CD11b⁺F4/80⁺) and M1 macrophages (F4/80⁺CD11c⁺), and attenuated the antigen-presenting capacity of macrophages. Notably, BBR combined with Sta was superior to BBR alone in reducing blood lipids and inhibiting the expression of hepatic pro-inflammatory genes Il-6. However, no significant difference was observed between the two groups in alleviating hepatic macrophage infiltration and improving oxidative stress indicators. Collectively, the combination of BBR and Sta exerts anti-MASLD (metabolic dysfunction-associated steatotic liver disease) effects by ameliorating hepatic lipid accumulation, inflammation, and oxidative stress, as well as reducing macrophage number and weakening its function. Furthermore, the effects of the combination in improving MASLD are better than BBR alone. These findings suggest that BBR combined with Sta is a potential effective strategy for the treatment of MASLD. The animal experiments were approved by the Institutional Animal Care and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College.

Malassezia furfur (M. furfur) is a common lipophilic fungus with strong epidermal colonization ability and high recurrence rates, capable of inducing a wide range of skin diseases in both animals and humans. Traditional antifungal therapies are associated with significant side effect and frequent drug resistance, necessitating the development of new treatment strategies. This study investigated the efficacy of a protonated phenanthroline-phenanthroline⁺ (ph-ph⁺) compound, previously developed by our team, in inhibiting the growth and reproduction of M. furfur. The results showed that the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of ph-ph⁺ against M. furfur were 2 and 8 µg·mL^-1, respectively. Furthermore, ph-ph⁺ significantly reduced key virulence characteristics of M. furfur, including biofilm formation, hydrophobicity, and adhesion capacity. Assessments using transmission electron microscopy and Annexin V-FITC/propidium iodide staining confirmed that ph-ph⁺ kills M. furfur by inducing apoptosis. In a mouse skin infection model induced by M. furfur, topical application of ph-ph⁺ effectively cleared the fungus from the skin surface without causing skin irritation or other adverse reactions. This study demonstrates that ph-ph⁺ may represent a novel antimicrobial compound distinct from traditional drugs and provides a potential therapeutic approach for treating skin infections caused by M. furfur or other fungi. The experimental protocol was approved by the Animal Experiment Ethics Committee of Southwest University (approval No.: LAC2024-1-0446), and the experiments were conducted strictly in accordance with the ethical principles for the use and care of animals.

This study aims to investigate the effect and mechanism of the chromone compound 19A derived from Saposhnikovia divaricata in ameliorating acute lung injury (ALI) in mice. Molecular docking, drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and dual-luciferase reporter gene assays were employed to elucidate the binding mode of 19A with farnesoid X receptor (FXR) and its regulatory effect on FXR transcriptional activity. In vitro, an inflammatory model was established by stimulating bone marrow-derived macrophages (BMDMs) with lipopolysaccharide (LPS), and the effects of 19A on cell viability, nitric oxide (NO) release, mRNA expression of inflammatory cytokines, and NF-κB p65 phosphorylation were evaluated. The FXR-dependence of these effects was verified in BMDMs derived from Fxr^-/- mice. In vivo, two ALI mouse models were established via intratracheal instillation of LPS and Escherichia coli (E. coli) to evaluate lung function, total protein concentration in bronchoalveolar lavage fluid (BALF), pulmonary inflammatory histopathology, inflammatory cytokine levels, and reverse validation was performed in Fxr^-/- mice. The use of animals in this study was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine, with the approval number: PZSHUTCM2412200007. 19A forms stable interactions with FXR, enhancing its transcriptional activity. In vitro, 19A inhibited LPS-induced NO release and the mRNA expression of inflammatory cytokines, reduced IκBα degradation and phosphorylation, and decreased NF-κB p65 phosphorylation; these anti-inflammatory and NF-κB-inhibitory effects were markedly attenuated or abolished in Fxr^-/- BMDMs. In wild-type mice, 19A markedly improved LPS-induced lung dysfunction, reduced lung index and BALF total protein, alleviated structural damage and inflammatory cell infiltration in lung tissue, and downregulated pulmonary inflammatory cytokine mRNA levels and NF-κB p65 phosphorylation. In contrast, these protective effects of 19A were largely lost in Fxr^-/- mice. In summary, the Saposhnikoviae Radix chromone 19A attenuates LPS-induced macrophage inflammatory responses and ALI in mice by activating FXR and inhibiting the NF-κB signaling pathway. Compound 19A may serve as a lead candidate for ALI prevention and treatment based on the chromone scaffold of Saposhnikoviae Radix, providing experimental support for the modernization of the traditional Chinese medicine Fangfeng and the development of new therapeutics for ALI.

Halorotetin B is a natural terpenoid compound isolated from the tunic of Halocynthia roretzi. Studies have shown that this compound exhibits significant anti-tumor activity. However, its anti-inflammatory potential remains unexplored, likely owing to the multi-target nature of natural compounds. In this study, we investigated the anti-inflammatory activity of Ha. B. The results showed that Ha. B significantly inhibited the production of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Mechanistically, network pharmacology analysis and Western blot assays verified that Ha. B exerts its anti-inflammatory function by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. Finally, natural product target prediction revealed that Ha. B may directly target mouse double minute 2 (MDM2). Cellular thermal shift assay (CETSA) results confirmed that Ha. B could interact with MDM2 in cells, and molecular docking was performed to simulate their physical binding mode. Meanwhile, RNA interference experiments further verified that Ha. B exerts its anti-inflammatory activity through binding to the E3 ubiquitin ligase MDM2.

Anxiety disorders affect over 3.3% of the global population, with limited treatment options available. Both trace amine-associated receptor 1 (TAAR1) and serotonergic 2C receptor (5-HT_2CR) are implicated in various emotional activities, including anxiety. This study aims to investigate the role of TAAR1/5-HT_2CR in the development of anxiety-like behavior in rats induced by chronic unpredictable mild stress (CUMS), and the intervention effects of a dual agonist. A CUMS-induced rat anxiety model was established. This animal research protocol has been approved by the Yantai University Laboratory Animal Ethics Committee (animal ethics number: YTU20240110). Anxiety-like behavior was assessed using the elevated zero-maze test and open field test. Western blot analysis was employed to detect the expression of TAAR1, 5-HT_2CR, and brain-derived neurotrophic factor (BDNF) proteins in the rat amygdala. To observe the effects of Taar1 and Htr2c gene on anxiety-like behavior in rats, we knocked down or overexpressed these genes in the rat basolateral amygdala (BLA). Results demonstrated that CUMS-induced anxiety significantly reduced the expression levels of TAAR1 and 5-HT_2CR in the amygdala of rats. Overexpression of either Taar1 or Htr2c in the BLA ameliorated anxiety-like behavior. Conversely, knocking down Taar1 or Htr2c in the BLA exacerbated CUMS-induced anxiety-like behavior. Oral administration of PCC0105004 alleviated anxiety-like behavior in CUMS rats, while knocking down both Taar1 and Htr2c in the BLA completely abolished this effect of PCC0105004. Further investigations revealed that PCC0105004 significantly increased the expression of BDNF in the amygdala of rats. These findings indicate that TAAR1 and 5-HT_2CR play crucial roles in the development of anxiety-like behavior in CUMS rats, suggesting that a dual agonist targeting TAAR1/5-HT_2CR may offer a novel therapeutic strategy for anxiety disorders.

The structural and functional homeostasis of the intestinal barrier plays a key role in the occurrence and progression of sepsis. As a highly selective α₂-adrenergic receptor agonist, dexmedetomidine is widely used for sedation in sepsis and perioperative periods requiring surgical intervention. Recent studies have shown that it also exerts anti-inflammatory effects; however, whether it can improve acute intestinal barrier injury in sepsis and the underlying mechanism remain unclear. In this study, an acute sepsis mouse model was established by intraperitoneal injection of lipopolysaccharide (LPS). Evans blue staining, transmission electron microscopy, HE staining, ELISA, and RT-PCR were used to observe the effects of dexmedetomidine (1-6 μg·kg^-1) on intestinal inflammation and barrier function in septic mice. The expression changes of α₂-adrenergic receptors during inflammation, as well as the effects of dexmedetomidine on NF-κB and RhoA/ROCK signaling pathways under LPS stimulation, were further evaluated in mouse intestinal tissues and RAW 264.7 macrophages. The results showed that dexmedetomidine (1-6 μg·kg^-1) administered 6 h after LPS stimulation significantly reduced the levels of inflammatory factors in plasma and intestinal tissues of septic mice, improved intestinal barrier function, and upregulated the expression of tight junction proteins in the small intestine and colon. LPS stimulation downregulated the expression of α₂-adrenergic receptors on macrophages in vivo and in vitro, whereas dexmedetomidine (5-20 nmol·L^-1) reversed this downregulation under inflammatory conditions, inhibited NF-κB pathway activation and inflammatory cytokine secretion in RAW 264.7 cells, and antagonized RhoA/ROCK pathway activation and decreased transepithelial electrical resistance in Caco-2 cells. In conclusion, this study demonstrates that LPS stimulation downregulates α₂-adrenergic receptor expression in macrophages, and dexmedetomidine can upregulate α₂-adrenergic receptors, thereby it may inhibit the NF-κB and RhoA/ROCK pathways, ultimately suppressing inflammation, improving intestinal barrier function, and alleviating acute intestinal barrier injury in sepsis. The animal experiments were approved by the Animal Ethics Committee of Southwest University (Ethics Review No.: IACUC-20260305-12).

This study investigates the protective effects of liquiritin (LQ) against acetaminophen (APAP)-induced liver injury in mice and explores the underlying mechanisms. A drug-induced liver injury model was established in mice by intragastric administration of APAP (300 mg·kg^-1). LQ was administered intragastrically at 1 h after APAP treatment. Serum biochemical indices, oxidative stress markers, and hepatic histopathological changes were evaluated at 6 h after APAP administration to comprehensively assess the ameliorative effects of LQ (animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine, approval Nos. PZSHUTCM2401080003 and PZSHUTCM2510170008). Furthermore, Western blot (WB), quantitative real-time polymerase chain reaction (qPCR), and in vitro cell models were employed to elucidate the underlying molecular mechanisms. The results demonstrated that LQ (25 and 50 mg·kg^-1) significantly ameliorated APAP-induced liver injury (P < 0.05, P < 0.01, P < 0.001), hepatic oxidative stress (P < 0.05, P < 0.01, P < 0.001) and alleviated the inflammatory response (P < 0.05, P < 0.01). WB and qPCR analyses revealed that LQ promoted the nuclear translocation and activation of nuclear factor erythroid 2-related factor 2 (Nrf2), and upregulated downstream gene expression in APAP-induced mouse livers (P < 0.05). Notably, the protective effects of LQ were abolished in Nrf2 knockout mice. These findings indicate that LQ alleviates APAP-induced hepatotoxicity primarily by activating Nrf2 antioxidant signaling pathway to counteract oxidative stress.

Cancer is the second leading cause of death worldwide, with solid tumors accounting for the majority of cases in adults. Advanced and metastatic solid tumors are responsible for most cancer-related deaths; however, current treatments still fail to achieve both high efficacy and low toxicity. In this study, we constructed an oncolytic virus (OV) named SKM_KK010(h) based on chimpanzee adenovirus serotype 23 (SAdV-23). This OV utilizes an optimized human telomerase reverse transcriptase (hTERT) promoter to drive e1a gene expression, thereby restricting viral replication and oncolysis to tumor cells. Additionally, it carries the csf2 transgene, which encodes human granulocyte-macrophage colony-stimulating factor (GM-CSF), to activate systemic anti-tumor immunity. Experiments in vitro demonstrated that SKM_KK010(h) selectively kills a broad range of tumor cell lines — including colon, bladder, liver, lung, and prostate cancer cells — while exhibiting no cytotoxicity toward normal cells. Viral replication in tumor cells was 100- to 1 000-fold higher than in normal cells. Furthermore, SKM_KK010(h) selectively and efficiently expresses bioactive GM-CSF in the tested tumor cells, reaching a level of 147.60 ± 7.63 ng per 10⁶ cells per 24 h (e.g., in LoVo cells), far exceeding the in vivo efficacy threshold (40 ng per 10⁶ cells per 24 h) required to elicit potent antitumor immunity. In contrast, GM-CSF expressions were undetectable or minimal in normal cells. In a LoVo xenograft mouse model, intratumoral injection of SKM_KK010(h) effectively inhibited tumor growth, achieving a relative tumor proliferation rate (T/C%) of 33.01%. In summary, we have successfully developed a safe and effective oncolytic chimpanzee adenovirus with strong potential as a platform for next-generation oncolytic virotherapy. All animal experiments were approved by the Institutional Ethics Committee and conducted in accordance with relevant ethical guidelines (Ethics Approval Number: IACUC-KYB2301P).

The rate constant of the bioorthogonal cleavage reaction is a critical parameter for its in vivo translation and application. Building upon the bioorthogonal decaging chemistry of silylborane and N-oxide, this study conducted structural optimization toward silylborane to obtain derivatives with higher second-order rate constants. The structure-reactivity relationships studies revealed that introducing electron-withdrawing groups onto the phenyl ring of the silylbenzyl moiety significantly enhanced the cleavage reaction rate. Furthermore, several silylboranes were successfully identified with fast reaction kinetics (k₂ > 10² L·mol^-1·s^-1). A representative derivative 2b (k₂ = 239 L·mol^-1·s^-1) was able to efficiently and specifically activate the N-oxide based fluorescent probe and prodrug of tamoxifen at the cellular level. In summary, this work provides an efficient cleavage reaction tool for the design of bioorthogonal prodrugs for in vivo applications.

ATP-citrate lyase (ACLY), serving as a critical energy metabolism bridge in tumor cells, is closely implicated in the development of various malignancies. Inhibition or knockout of ACLY has been reported to effectively suppress tumor cell proliferation and induce apoptosis. In this work, we have designed and synthesized 18 dehydrocurvularin cinnamy ACLY inhibitors. Through biological evaluation and molecular dynamics simulations, we systematically explored the structure-activity relationships (SAR) of these derivatives. Compound 4e exhibited potent anti-proliferative activity against MDA-MB-231 and PANC-1 cells with a IC₅₀ value of 0.4 and 1.1 μmol·L^-1, providing crucial insights for structural modification and SAR studies of this class of macrolide derivatives.

A systematic phytochemical investigation of Polygonatum cyrtonema Hua. processed by nine cycles of steaming and sun-drying was carried out using silica gel, Toyopearl HW-40C, octadecylsilyl (ODS) silica gel, and semi-preparative HPLC, leading to the isolation of three nitrogen-containing compounds. The structures of the isolates were elucidated by comprehensive spectroscopic analyses, including HRESIMS, NMR, IR, and UV, and were identified as cyrtoneine F (1), (2S,5S)-5-[(1S)-1-methylpropyl]-3,6-dioxo-2-piperazine propanoic acid (2), (2S,5S)-5-(2-methylpropyl)-3,6-dioxo-2-piperazine propanoic acid (3). Compound 1 was characterized as a new amide, whereas compounds 2 and 3 were determined to be diketopiperazines, which were isolated for the first time from Polygonatum cyrtonema Hua.

The secondary metabolites of the fungus Furcasterigmium sp. L-YB-P1-1, isolated from Jinlun Cave in Guangxi were investigated. The crude extract was separated and purified by chromatographic techniques including thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The structures of the isolated compounds were elucidated through comprehensive spectroscopic analysis such as HNMR and mass spectrometry. Ten compounds were obtained, comprising one new compound, (2R,3S)-2-hydroxy-3-methylpentanoic acid (E)-5′-hydroxy-3′-methylpent-2′-enoate (1) and 9 known compounds: Uridine (2), adenosine (3), 2-methoxyl-β-L-arabinofuranosyl uracil (4), thymidine (5), 2′-O-methyladenosine (6), 5′-O-acetyl-9-β-arabinofuranosyl adenosine (7), uridine-5′-α-hydroxypropanoate (8), asperteretal G (9), and trans-ferulic acid (10). The antioxidant activity of the compounds was evaluated using the DPPH free radical scavenging assay, which revealed that compounds 2-5 and 10 possessed antioxidant activity. Furthermore, the anti-inflammatory activity was assessed by measuring the inhibition of NO production in LPS-induced RAW 264.7 cells using the Griess method, indicating that compound 9 exhibited certain anti-inflammatory activity.

Two compounds were isolated from the traditional Chinese medicine Ganoderma lucidum using various chromatographic techniques. Through methods such as one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy, both compounds were identified as new lanostane-type triterpenoids. Their absolute configurations were determined via quantum chemical calculations, and they were named baoslingzhines X and Y (1 and 2). In vitro and in vivo experimental results revealed that these two compounds exhibit potent activity against neuroinflammation, oxidative stress, and depression. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Shenzhen University (approval number: IACUC-202400148).

Verticillium-derived secondary metabolites have demonstrated promising biological activities, including antibacterial, antitumor, and antimalarial effects. To further explore the chemical diversity and bioactive potential of secondary metabolites from the genus Verticillium, the present study aimed to isolate and characterize secondary metabolites from the fermentation broth of the fungal strain Verticillium sp. SN-1 isolated from Daya Bay, Guangdong Province, China and to evaluate their antibacterial activities. Thirteen compounds were isolated from the crude extract of Verticillium sp. SN-1 using a combination of chromatographic techniques, including normal-phase silica gel column chromatography, reversed-phase ODS column chromatography, and semi-preparative high-performance liquid chromatography (HPLC). Their chemical structures were elucidated by comprehensive spectroscopic analyses, such as nuclear magnetic resonance (NMR) spectroscopy, ultraviolet (UV) spectroscopy, and mass spectrometry (MS). Based on spectral data interpretation, the compounds were identified as cytosporone G (1), verticillpyrone A (2), methyl 1, 5-dihydroxy-3-methylxanthone-6-carboxylate (3), acremonidin E (4), acremonoxide (5), alternethanoxin A (6), janthinone (7), 3-deoxy-methylsulochrin (8), astrophenone (9), acremoxanthone F (10), acremoxanthone C (11), acremoxanthone B (12), and acremonidin G (13). Among these isolates, compounds 1 and 2 were characterized as new compounds, while compounds 3-13 were known secondary metabolites. The antibacterial activities of all isolated compounds were evaluated using the broth microdilution method according to established protocols. The results showed that compounds 4 and 10-13 exhibited significant inhibitory effects against methicillin-resistant Staphylococcus aureus (MRSA), with minimum inhibitory concentrations (MIC) of 32, 16, 32, 8, and 8 μg·mL^-1, respectively. Additionally, compounds 11-13 displayed inhibitory activities against Enterococcus faecalis, with corresponding MIC values of 32, 32, and 8 μg·mL^-1.

Six new compounds including two diarylheptanoids (1, 2), one norlignan (3), and three gingerenols (4-6) were isolated from the rhizoma of Zingiber officinale by column chromatography of Diaion HP-20, MCI Gel CHP-20, Sephadex LH-20, ODS-A gel and semi-preparative HPLC. Structural identification of the isolated compounds was performed via modern spectroscopic methods (1D and 2D NMR, UV, IR, and MS), and they were characterized as 3-keto-5(R)-(6-amino-9H-purin-9-yl)-1, 7-(4-hydroxy-3-methoxyphenyl)-heptane (1), (5α, 6α)1, 5-epoxy-3-one-6-hydroxy-1, 7-(4-dihydroxy-3-methoxyphenyl)-heptane (2), (2S)1-(bis(4-hydroxy-3-methoxyphenyl)methyl)-dihydrofuran-5-(2H)one (3), 1-(4-hydroxy-3-methoxyphenyl)-decane-3, 5, 6-triol (4), 1-(4-hydroxy-3-methoxyphenyl)-decane-3, 8-diol (5) and 1-(4-hydroxy-3-methoxyphenyl)-decane-3, 5, 10-triol (6), by using modern spectroscopic methods (1D and 2D NMR, UV, IR, and MS). The absolute configurations of compounds 1 and 3 were determined through the comparisons of optical rotations in references. However, the ECD data of compound 3 were failed to obtain, and only the relative configuration was determined through the NMR calculations. Due to insufficient sample amounts, the absolute configurations of compounds 3-6 were not further determined.

A total of seven furan natural products were isolated from the ethyl acetate extract of the fermentation product of the mycelium of Cordyceps militaris through a combination of chromatographic techniques, including silica gel column chromatography and semi-preparative HPLC. Based on their physicochemical properties and spectroscopic data, the structures of isolated compounds were identified as ethyl 5-[(3R)-hydroxybutyl]-2-furanacetate (1), 5-[(3R)-hydroxybutyl]-2-furancarboxylic acid (2), methyl 5-[(3R)-hydroxybutanoyl]-2-furanacetate (3), 5-[(3R)-hydroxybutyl]-2-furancarboxaldehyde (4), methyl 5-(2-hydroxyethyl)-2-furanacetate (5), 5-[(3R)-hydroxybutyl]-2-furanacetic acid (6), and methyl 5-[(3R)-hydroxybutyl]-2-furanacetate (7). Among them, compounds 1-3 were new compounds, compound 4 and 5 represents a new natural product, and compound 6 and 7 were isolated from Cordyceps militaris for the first time. Compounds 1 and 3-7 exhibited anti-inflammatory activity in varying degrees and significantly inhibited nitric oxide production in lipopolysaccharide-induced mouse mononuclear macrophage RAW264.7 cell line. Compound 2 showed some cytotoxic activity against Spodoptera frugiperda ovarian Sf9 cells with an IC₅₀ value of 57.01 μmol·L^-1.

The purpose of this study was to evaluate the pharmacokinetic behavior and metabolite analysis of a novel anti-triple negative breast cancer candidate compound MT-1 in SD rats. Rats were randomly divided into intragastric gavage group and tail vein injection group. Blood samples were collected at different time points after single dose of MT-1 administration. An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established to determine the concentration of MT-1 in plasma with coumarin as the internal standard, and the methodology was verified. The pharmacokinetic parameters were calculated by DAS 2.0 software, and the blood concentration-time curve was drawn by GraphPad Prism 10.1.2 software. At the same time, UPLC-Q-TOF/MS^E combined with UNIFI software was used to analyze the metabolites of plasma samples. The results of pharmacokinetic analysis showed that the oral bioavailability of MT-1 was 11.26%, and the absorption was fast. The quantitative range of methodological investigation was 2-800 ng·mL^-1, showing a good linear relationship. The matrix effect, extraction recovery, accuracy and stability all met the standards of biological analysis. Metabolite analysis speculated four potential metabolites, mainly involving methylation, cysteine and glucuronic acid binding. This study systematically revealed the pharmacokinetic characteristics and metabolic transformation of MT-1 in vivo, which provided an experimental basis for its further development and application in the treatment of triple-negative breast cancer. This experiment was approved by the Experimental Animal Ethics Committee of Guizhou Medical University. (approval No.: 2503423).

Psoralea corylifolia L., a widely used traditional Chinese medicine, has recently demonstrated therapeutic potential in the treatment of liver cancer. However, rigorous evaluations regarding its clinical efficacy and quality control remain insufficient. To address this issue, this study selected 10 batches of Psoralea corylifolia L. from different habitats as research objects. Firstly, high-performance liquid chromatography (HPLC) was employed to optimize the chromatographic conditions, and based on this, the chemical fingerprints of the 10 batches of medicinal materials were constructed to characterize their chemical composition profiles. Meanwhile, the IC₅₀ values of medicinal extracts from each batch against Hepa1-6 liver cancer cells were determined; using spectrum-effect correlation analysis, potential anti-liver cancer active components were preliminarily screened, and their activities were further validated by cell experiments. Finally, an effective-constituents index (ECI) was established for the 10 batches of Psoralea corylifolia L. and applied to its quality evaluation. The results showed that although the similarity of the chemical fingerprints of the 10 batches of Psoralea corylifolia L. exceeded 0.95, indicating a relatively consistent overall chemical composition, the content of common components varied significantly among different batches. In terms of anti-liver cancer activity, the IC₅₀ values of the 10 batches of medicinal materials against Hepa1-6 cells ranged from 34.54 μmol·L^-1 (minimum) to 85.04 μmol·L^-1 (maximum), showing obvious differences in activity, which further confirmed the necessity of establishing a more targeted quality evaluation system. Through spectrum-effect relationship analysis and in vitro cell activity verification, it was confirmed that isobavachin, neobavaisoflavone, bavachin, psoralidin, isobavachalcone, bavachinin and bakuchiol are the main active components responsible for the anti-liver cancer effect of Psoralea corylifolia L. Notably, psoralen and isopsoralen, which are specified as index components in the Pharmacopoeia, showed no inhibitory activity against Hepa1-6 liver cancer cells in this study. In addition, the constructed ECI could classify the 13 batches of Psoralea corylifolia L. into three quality grades, and this classification result was consistent with the results of linear discriminant analysis, indicating good reliability of the ECI. More importantly, the correlation coefficient between the ECI of Psoralea corylifolia L. and its anti-liver cancer activity reached as high as 0.957, which fully proved that the ECI can effectively reflect the anti-liver cancer efficacy of Psoralea corylifolia L. In conclusion, the ECI established in this study can be used for the quality evaluation of the anti-liver cancer effect of Psoralea corylifolia L. It not only provides a new method for improving the quality control standard of Psoralea corylifolia L. but also offers a valuable reference for the quality control and efficacy evaluation of other anti-tumor traditional Chinese medicines.

To investigate the in-situ spatial distribution and dynamic transformation of ginsenosides and lipids during the steaming process of Panax ginseng, desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) was employed to analyze the main root sections steamed for 0, 2, 4, 6, 8, and 10 h in both the positive and negative ion modes. Ginsenosides were detected using 95% methanol containing 0.1 mmol·L^-1 NH₄Cl as the spray solvent, while lipids were detected using methanol containing 0.01 mmol·L^-1 TFA. The spray solvent flow rate was 2 μL·min^-1, with X and Y pixel sizes of 200 μm and a raster speed of 500-700 μm·s^-1. The results showed that, after the steaming, most macromolecular ginsenosides underwent degradation reactions such as demalonylation, deglycosylation, and deacetylation, leading to an increase in the content of rare ginsenosides. With prolonged steaming time, the signal intensity of most lipids decreased, and their distribution contours became blurred. Through multivariate statistical analysis, a total of 29 differential markers were screened and identified, including seven ginsenosides and 22 lipids, which were mainly distributed in the epidermis, cortex, and xylem, and their content changes were closely correlated with the steaming time. DESI-MSI technology can intuitively reveal the spatial distribution and dynamic transformation patterns of chemical components during the steaming process of P. ginseng, and the screened markers provide new scientific basis for studying the processing mechanisms and quality evaluation of P. ginseng.

Recombinant adeno-associated virus (rAAV) is a pivotal vector for gene therapy, and accurate identification of its capsid proteins is crucial for ensuring product safety, efficacy, and batch-to-batch consistency. Although existing LC-MS peptide mapping methods based on in-solution digestion can confirm the primary structure of proteins, they suffer from an inherent limitation: VP1, VP2, and VP3 share an identical C-terminal sequence (over 400 amino acids in the case of AAV9). Consequently, common peptides generated from mixed digestion cannot be assigned to their parental protein origin, making it particularly difficult to independently assess the sequence coverage integrity and specific post-translational modifications of low-abundance VP1 and VP2. To address this inherent limitation, this study establishes, for the first time, a systematic peptide mapping method for rAAV capsid proteins based on "SDS-PAGE separation, in-gel digestion, and LC-MS analysis". The core strategy of this method involves the initial physical separation of the three capsid proteins (VP1, VP2, and VP3) by electrophoresis, followed by independent in-gel trypsin/chymotrypsin digestion of each individual protein band, thereby enabling specific evaluation of the sequence coverage for each protein. It is noteworthy that despite the natural stoichiometry of VP1, VP2, and VP3 in the capsid being approximately 1:1:10 (i.e., VP1 and VP2 each constitute only about 1/12 of the total protein), and despite further loss of peptide quantities during the multi-step recovery process of in-gel digestion, this method achieves a combined sequence coverage of over 98% for the low-abundance VP1 and VP2, fully demonstrating its exceptional detection sensitivity and specificity. Method validation demonstrated that the approach is highly specific (no interference from blank matrix), reproducible (total peptide coverage ≥ 98% in triplicate runs, retention time RSD of key peptides < 1%), and robust (stable coverage under variations of key parameters). This method effectively resolves the challenge of ambiguous peptide assignment caused by identical C-terminal sequences of VP1/VP2/VP3 in solution-based digestion, providing a reliable, stable, and high-resolution analytical tool for accurate serotyping, quality attribute investigation, and quality control of rAAV products.

Due to the inability of the current quality standards to effectively control the quality of Lepidii Semen, this study established the HPLC characteristic chromatogram and multi-component content determination method for Lepidii Semen. Using a NanoChrom ChromCore HP AQ C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile and 0.2% phosphoric acid aqueous solution as the mobile phase for gradient elution at a flow rate of 1.0 mL·min^-1, with a column temperature of 35 ℃, and detection wavelength of 210 nm. The characteristic HPLC chromatogram of Lepidii Semen was established with twelve characteristic peaks marked, eleven of which were identified by comparison with reference standards. The similarity of 13 batches of market-sourced samples exceeded 0.90. The established chromatogram effectively distinguished Lepidii Semen from four other cruciferous seeds and four common adulterants. The method was simultaneously applied to determine the contents of glucotropaeolin (GPL), glucoerucin (GR), sinapine thiocyanate (SPT), and quercetin-3-O-β-D-[2-O-(6-O-sinapoyl)-β-D-glucopyranosyl]-glucopyranoside (QSGG). Two detection wavelengths (210 and 330 nm) were selected based on the maximum absorption of each compound. Method validation confirmed that the method possesses high specificity, good linearity, and satisfactory precision, repeatability, stability, and recovery rates. The contents range of GPL, GR, SPT and QSGG in 13 batches of Lepidii Semen were 1.58%-2.40%, 1.27%-1.78%, 0.22%-0.27%, and 0.28%-0.42%, respectively. The established HPLC characteristic chromatogram and multi-component content determination methods are stable and reliable, providing a scientific basis for the quality control and evaluation of Lepidii Semen, standardization of the herbal market and rational clinical use.

Gallbladder cancer (GBC) is a highly invasive malignant tumor of the digestive system. Chemotherapy plays a crucial role in its advanced treatment, yet significant interpatient variability in drug response and prominent drug resistance issues persist. Currently, there is a lack of systematic transcriptomic databases and drug response prediction tools specifically tailored for GBC. To address this gap, we developed the Gallbladder Cancer Database (GBCDB, http://www.gbcdb.com/). This platform integrates five bulk-transcriptome datasets (126 samples) and one single-cell dataset (24 samples, encompassing 140 870 cells across eight major cell lineages), comprehensively covering neoplastic tissues, adjacent normal tissues, and cholecystitis tissues. GBCDB offers a user-friendly interface supporting flexible query modes for single-gene, dual-gene, and multi-gene analyses. The platform enables comprehensive comparative analysis through multiple visualization tools: Box plots display expression distributions across sample types, correlation scatter plots reveal gene associations, heatmaps visualize gene expression patterns, and Unified Manifold Approximation and Projection (UMAP) is applied to single-cell transcriptomic data. Additionally, the RefDNN model predicts resistance to five commonly used gallbladder cancer chemotherapeutics — gemcitabine, cisplatin, 5-fluorouracil, mitomycin C, and paclitaxel. GBCDB provides systematic data resources and analytical tools for molecular mechanism research and chemotherapy resistance assessment in gallbladder cancer, offering scientific foundations for developing personalized treatment strategies.

To evaluate the excretion pathway, pharmacokinetic properties, and cardiac safety of the dipeptidyl peptidase-4 (DPP-4) inhibitor LGT-6 in rats, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was used to determine drug concentrations in biological samples, in combination with an automated patch-clamp technique (automated patch clamp, APC) to assess the risk of human ether-à-go-go-related gene (hERG) potassium channel inhibition. The results showed that LGT-6 is primarily excreted via the biliary route in rats, with a cumulative excretion rate of approximately 82.4%. Following single-dose administration via tail vein injection (5 mg·kg^-1) or oral gavage (5 mg·kg^-1), the absolute bioavailability of LGT-6 was 77.2%. The IC₅₀ of LGT-6 against the hERG potassium channel was greater than 30 μmol·L^-1. The analytical methods established in this project for quantifying various target components in in vivo and in vitro biological samples all complied with relevant regulatory technical requirements. These data indicate that LGT-6 exhibits absorption, distribution, metabolism, and excretion (ADME) characteristics dominated by biliary excretion and high oral absorption efficiency, along with a low cardiac safety risk, thereby providing key experimental evidence to support its further preclinical safety evaluation and subsequent development. This experiment was approved by the Animal Ethics Committee of Guizhou Medical University (Grant No: 1900912).

The study aims to investigate the metabolic process of the antidepressant component (BS-60E) from Paeoniae Radix Alba in rats, and to compare the differences in the disposition of its metabolites between normal and depressed states. In this study, a chronic unpredictable mild stress (CUMS) depression model was established in rats. Normal and depressed rats were orally administered the BS-60E aqueous solution, and serum and urine samples were collected. The prototype constituents and metabolites were identified using UPLC-Q-TOF/MS combined with UNIFI software, and multivariate statistical analysis was employed to compare the differential constituents between normal and depressed rats. Animal experimental procedures were approved by the Animal Ethics Committee of the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College (approval No.: SLXD-20211105019). The results showed that 9 and 28 prototype constituents, as well as 33 and 60 metabolites, were detected in serum and urine samples, respectively. The main metabolic pathways included phase Ⅰ reactions such as hydroxylation and desaturation, and phase Ⅱ reactions including glucuronidation and sulfation. Compared with normal rats, the levels of monoterpene constituents such as paeoniflorin and 1-O-β-D-glucopyranyl paeoniflorone were decreased in both serum and urine of depressed rats, while the levels of polyphenolic constituents such as methyl gallate and ethyl gallate were decreased in serum but increased in urine. These results indicate that the depressed state significantly alters the metabolic disposition of the main constituents of BS-60E in rats. This is manifested as reduced absorption of monoterpene constituents, leading to decreased exposure of prototype constituents in serum and urine, suggesting that their pharmacological effects may primarily depend on metabolites and are excreted via the kidneys in metabolized forms. Meanwhile, the renal excretion of polyphenolic constituents is accelerated, resulting in decreased serum concentrations and increased urinary excretion, suggesting that their prototype constituents may directly participate in the pharmacological process and are excreted in urine both as prototypes and metabolites.

To explore a method for the selective separation and enrichment of protocatechuic acid (PCA) from actual samples of traditional Chinese medicinal materials, in this study, based on the boron affinity and molecular imprinting strategy, a specific imprinted magnetic material with high affinity for PCA was prepared by using Fe₃O₄ magnetic nanoparticles (Fe₃O₄ MNPs) as the substrate and amplifying the number of boronic acid ligands on the product surface with branched polyethylenimine (PEI). The prepared Fe₃O₄@PEI@FPBA MIPs were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), and other instruments, and their adsorption performance was systematically investigated. Due to the boron affinity-directed surface imprinting mechanism, the prepared Fe₃O₄@PEI@FPBA MIPs showed high selectivity for PCA, with a maximum adsorption capacity of 475.33 mg·g^-1 and a maximum imprinting factor (IF) of 5.24. The adsorption process conformed to the pseudo-first-order kinetic model, and the adsorption isotherm conformed to the Langmuir model. In addition, the results of the repeatability test also indicated that Fe₃O₄@PEI@FPBA MIPs could maintain good adsorption performance after repeated use. The recovery rate of the actual sample analysis was 96.67% to 110.17% (relative standard deviation was 1.65% to 4.52%).

Local precision therapy for osteoarthritis (OA) still faces dual challenges of low delivery efficiency and high toxicity. This study constructed a triptolide liposome microneedles (TP-Lip-MNs) delivery system. TP liposomes (TP-Lip) were prepared by the thin-film dispersion-ultrasonication method, and the microneedle preparation process was optimized via microneedle mold casting-centrifugal filling-drying technology. The final TP-Lip-MNs exhibited a drug loading content of 63.49 μg per needle, a single-needle mechanical strength of 0.9 N, and a peak drug release at a skin depth of 120 μm. In the pharmacokinetic (PK) study, a quantitative analysis method for TP in rat knee joint microdialysate was established using microdialysis sampling combined with LC-MS/MS. The results showed that the area under the curve (AUC_{0-8 h}) of the TP-Lip-MNs group was 156.80 ± 37.04 ng·h·mL^-1, 2.3-fold higher than that of the TP-Lip group (68.31 ± 5.92 ng·h·mL^-1), and the peak concentration (C_max) was (66.35 ± 18.59 ng·mL^-1), 3.0-fold higher than that of the TP-Lip group (22.49 ± 2.36 ng·mL^-1), significantly enhancing the drug diffusion and absorption rates of TP in the knee joint. In the pharmacodynamic (PD) evaluation using a sodium monoiodoacetate-induced rat OA model, TP-Lip-MNs inhibited inflammation-induced bone hyperplasia as demonstrated by knee swelling rate and CT imaging. The treatment group showed a significant decrease in IL-1β, IL-6, and TNF-α levels after administration, further verifying the anti-inflammatory effect of TP. Serum biochemical indices (ALT, AST, BUN, CRE) confirmed its good biosafety. This study provides an innovative and translationally potential solution for local OA therapy through a "model-delivery-PK-PD" closed-loop design. The animal experiments were approved by the Ethics Committee for Animal Experiments of Tianjin University of Traditional Chinese Medicine under the ethical review number TCM-LAEC2024125g1009.

This study used functionality-related characteristics of disintegrants to clarify how different superdisintegrants act in traditional Chinese medicine (TCM) tablets and how their effects are modified by TCM type and drug loading. Three commonly used superdisintegrants and three TCMs representing disintegrating, dissolving/eroding and swelling disintegration patterns were selected as models. The basic properties of the disintegrants, including particle size, moisture, swelling ability and flowability, were systematically characterized and related to tablet disintegration profiles obtained from dynamic disintegration tests. The results showed that the performance of croscarmellose sodium was mainly governed by powder flowability, whereas sodium starch glycolate was controlled by a combination of sedimentation behaviour, particle size and moisture, and crospovidone depended primarily on particle size characteristics and swelling capacity. In tablets containing disintegrating- or dissolving/eroding-type TCMs, selecting grades according to these key properties clearly shortened disintegration time and increased disintegration rate. In high-loading tablets containing swelling-type TCM, competition for water and swelling space between the herbal powder and the disintegrant was observed and weakened the accelerating effect of the disintegrant. These findings indicate that an evaluation framework centred on functionality-related characteristics can link the structure and properties of superdisintegrants to disintegration behaviour in TCM tablets and provides a practical basis for optimizing disintegrant type and grade, particularly in high-loading formulations.

Chinese Medicine Carbon Dots, as an innovative achievement integrating nanotechnology and traditional Chinese medicine in recent years, exhibit unique advantages such as high efficiency, excellent water solubility, and outstanding biocompatibility, demonstrating significant potential in clinical therapy and other areas. This study utilized traditional Chinese herbs Eclipta prostrata L. and Ligustrum lucidum Ait. as precursors to synthesize Eclipta-Ligustrum carbon dots (EL-CDs) via a solvothermal method, exploring their potential as a novel agent for the prevention and treatment of breast cancer. Initially, the carbon dots were characterized using fluorescence spectroscopy and ultraviolet-visible (UV-Vis) absorption spectroscopy, confirming that EL-CDs possess favorable optical properties. The synthesis conditions were optimized to identify the optimal reaction parameters. To further investigate the characteristics of the carbon dots, mouse breast cancer 4T1 cells were used as the model system. Cellular imaging techniques demonstrated that EL-CDs can be effectively applied for cell imaging. MTT assays indicated that EL-CDs exert cytotoxic effects on 4T1 cells, while scratch assays revealed their ability to inhibit the migration of 4T1 cells. The changes in reactive oxygen species (ROS) levels in 4T1 cells were detected using DCFH-DA staining combined with inverted fluorescence microscopy. Compared to the control group, the EL-CDs treatment group showed a significant increase in ROS expression in a dose-dependent manner. To evaluate the in vivo imaging potential of EL-CDs, both in vivo and ex vivo organ imaging in mice demonstrated their excellent fluorescence properties. Furthermore, EL-CDs exhibited certain antitumor effects without significantly affecting the body weight or growth of the mice during treatment. In summary, EL-CDs possess good biocompatibility, remarkable fluorescence properties, and certain antitumor efficacy, offering a new strategy for breast cancer treatment and holding potential value for clinical translation. This experiment was approved by the Animal Ethics Committee of Shandong University of Traditional Chinese Medicine (No: SDUTCM20240304213).

It is difficult for anti-tuberculosis drugs to reach the tuberculous cavity lesions, and the effects of conventional oral or intravenous administration are not satisfactory. In this study, a rifapentine composite thermosensitive gel with poly (lactide-co-glycolide)-block-poly (ethylene glycol)-block-poly (lactide-co-glycolide) (PLGA-PEG-PLGA) as the gel matrix was developed for bronchoscopic pulmonary interventional therapy of pulmonary tuberculosis to enhance the targeted drug delivery treatment effect for pulmonary tuberculosis. An optimal formulation for rifapentine lipid microspheres was screened via orthogonal design. The lipid microspheres were then mixed with PLGA-PEG-PLGA solution to prepare a composite thermosensitive gel. The formulation was then characterized. The erosion of the in-situ gel was evaluated using a membrane-free method, while the in-vitro release and release mechanism were assessed by dialysis bag method. Uptake of the drug in the gel by A549 cells was observed using confocal microscopy. Antibacterial assays were assessed using lung tissue homogenates collected at various time points following a single administration, in order to investigate the pulmonary retention of the thermosensitive gel after intratracheal instillation in rats. All animal experimental procedures were approved by the Experimental Animal Ethics Committee of Hebei Medical University (No: IACUC-Hebmu-P 2022084) and carried out in accordance with its guidelines. The particle size of the prepared lipid microspheres was 482.4 ± 0.22 nm, which increased slightly after incorporation into the composite gel. The gelation temperature of the composite gel was 34.47 ± 0.15 ℃, with a gelation time of 41.33 ± 1.70 s. The formulation exhibited good injectability before gelation while showed a suitable viscosity at gelation temperature. The drug release model fitted with the Ritger-Peppas equation. The results of laser confocal microscopy showed that the lipid microspheres effectively delivered lipophilic drugs into lung epithelial cells. Antibacterial testing of lung tissue extracts confirmed that a single dose of the composite gel could sustain antibacterial activity in rat lungs for at least 14 days. The rifapentine lipid microsphere-thermosensitive gel composite formulation significantly enhances the solubility of rifapentine. Its temperature-dependent phase transition from liquid to gel not only ensures feasibility for bronchoscopic administration but also enables prolonged and effective drug release at the lesion site, thereby achieving the design goal of efficient local therapy for tuberculosis.

As a traditional bulk Chinese medicinal herb, Salvia miltiorrhiza's phenolic acids are the core bioactive components responsible for its pharmacological effects. Environmental stress can regulate relevant metabolic pathways to promote the accumulation of phenolic acids. To identify key transcription factor genes in S.miltiorrhiza under H₂O₂ stress, we first used aseptic seedlings cultured under different H₂O₂ concentrations as experimental materials, performed RNA-seq analysis following standard protocols with three biological replicates, and conducted untargeted metabolomics analysis via UHPLC-QE-MS with six biological replicates. All assays were commissioned to Gene Denovo Biotechnology Co., Ltd. The results showed a concentration-dependent stress effect: Low concentrations of H₂O₂ slightly inhibited seedling growth but activated genes associated with photosynthesis and phenolic acid biosynthesis. In contrast, high concentrations of H₂O₂ strongly suppressed growth and redirected metabolic resources from growth to defense processes. This response was accompanied by the upregulation of key enzyme genes in the phenylpropanoid and tyrosine pathways, including PAL, RAS and CYP98A2, thereby promoting the accumulation of ferulic acid (FA), rosmarinic acid (RA) and salvianolic acid B (Sal B). Meanwhile, oxidative stress repressed the expression of genes related to phytohormone synthesis and photosynthesis, reducing resource allocation for growth, and maintained intracellular redox homeostasis by activating genes such as GRXC1. Transcriptional regulatory analysis revealed that WRKY28 expression was significantly and positively correlated with salvianolic acid content, while MYB305 was inhibited as a negative regulator. These findings elucidate the regulatory network underlying phenolic acid accumulation in S. miltiorrhiza under H₂O₂ stress from the perspectives of metabolic resource allocation and transcriptional regulation, providing a preliminary theoretical basis for improving the quality of S. miltiorrhiza medicinal materials through molecular breeding or environmental regulation. It should be noted that exogenous H₂O₂ treatment is a controllable experimental method, and its conclusions may differ from those under natural stress in terms of signal initiation and network integration. Further verification through multi-dimensional approaches is required for refinement.

Bupleurum giraldii (H. Wolff) Koso-Pol. is a perennial herbaceous plant belonging to the Bupleurum L. (Apiaceae). It is recognized as a provincially protected rare medicinal plant in Shaanxi with the whole plant has acknowledged medicinal value. In this study, the complete plastid genome sequence of B. giraldii was sequenced and characterized, and comparative and phylogenetic analyses were then conducted alongside other published Bupleurum plastid genome sequences. The whole plastid genome of B. giraldii was 155 761 bp in length, which exhibited the conserved circular quadripartite structure consisting of a large single copy (LSC, 85 663 bp), a small single copy (SSC, 17 492 bp), and a pair of inverted repeats (IRa/IRb, 26 303 bp). A total of 129 genes were annotated, including 84 protein-coding genes (PCGs), 37 transfer RNA (tRNA) genes and 8 ribosomal RNA (rRNA) genes, of which, 6 protein-coding genes, 7 tRNA genes and 4 rRNA genes were duplicated in the IR regions. A total of 66 simple sequence repeats (SSRs), 34 tandem repeats, and 31 dispersed repeats were detected. In the plastid genome of B. giraldii, codons with the relative synonymous codon usage (RSCU) value greater than 1 showed a tendency to end in adenine (A) or thymine (T), whereas those with an RSCU value less than 1 tended to end in cytosine (C) or guanine (G). Collectively, neutral plotting, ENC-plot, and PR2-plot analyses all revealed that natural selection might be the predominant factor shaping this codon usage bias. The plastid genome of Bupleurum species exhibited highly conserved gene order and content, along with little variation in overall size and GC content, no significant expansion or contraction of the IR regions was detected. Interspecific sequence variation mainly occurred in the non-coding intergenic regions, and sequence divergence in the LSC and SSC regions was significantly higher than that in the IR regions. Eleven divergence hotspot sequences were identified, located in the LSC and SSC regions respectively. Phylogenetic inferences inferred that all Bupleurum species clustered into a monophyletic clade with BS_ML = 100%, PP = 1, and which could be further divided into two independent lineages corresponding to subgenus Bupleurum and subgenus Penninervia. Bupleurum giraldii located in subgenus Bupleurum and showed the closest relationship to B. falcatum L. The complete plastid genome of B. giraldii was comprehensively investigated for the first time and its phylogenetic position in Bupleurum was elucidated with high resolution in this study, these results will provide basis for the conservation and sustainable utilization of the germplasm resources of medicinal Bupleurum species.

Agarwood is an aromatic resin produced by Aquilaria sinensis and other plants of the Thymelaeaceae family in response to external injury. It is valued both medicinally and as a fragrance material. Sesquiterpenoids are key components responsible for the characteristic aroma and bioactivity of agarwood, but their biosynthetic pathway remains incompletely elucidated. In this study, three sesquiterpene synthase genes AsTPS2, AsTPS10, and AsTPS41, were cloned and identified from methyl jasmonate (MeJA)-induced callus of A. sinensis. Recombinant proteins were obtained by expression and purification in Escherichia coli. In vitro enzymatic assays revealed that AsTPS2 and AsTPS10 exhibit similar functions: Both catalyze farnesyl diphosphate (FPP) to produce α-humulene and β-caryophyllene, and also convert geranyl diphosphate (GPP) into acyclic monoterpenes (geraniol and nerol) as well as monocyclic monoterpenes (α-terpineol and isopulegol). AsTPS41 catalyzes FPP to form β-elemene, α-guaiene, and δ-guaiene, while also acting on GPP to generate geraniol, nerol, and α-terpineol. Protein modeling and docking of AsTPS2 and AsTPS10 using AlphaFold3 and AutoDock Vina indicated that, despite sharing 83.5% sequence identity, residues lining their active cavities are completely identical, supporting their overlapping catalytic activities. Further site-directed mutagenesis was performed on key amino acids lining the active cavity of AsTPS2. The single-site mutant Y411A generated two new products, β-elemene and shyobunol, while retaining the functionality of AsTPS2. Building upon Y411A, the double-site mutant G410A-Y411A produced four additional new products: α-Elemol, nerolidol, trans-farnesol, and pogostol. In the single-site mutant C449A, the proportion of β-caryophyllene increased from 7.2% in wild-type AsTPS2 to 55.1%. Based on these findings, a Saccharomyces cerevisiae expression system was established for AsTPS2, its mutant AsTPS2^C449A, and AsTPS41. Following isolation and structural analysis by nuclear magnetic resonance (NMR), the products α-humulene, β-caryophyllene, and δ-guaiene were identified. These results provide insights into the molecular mechanisms underlying agarwood formation and lay a foundation for further research on enzymes involved in the post-modification steps of agarwood sesquiterpenoid biosynthesis.

Lonicera japonica is a well-known traditional medicinal plant in China, and its floral volatiles are also used in the food, cosmetic, and horticultural industries. The artificially bred variety "Jiu Feng 2" is widely cultivated as a raw material for herbal teas and health products. However, its aromatic profile and underlying biosynthetic basis remain poorly understood. Here, we performed comparative volatile metabolomic and transcriptomic analyses of "Jiu Feng 2" and its parental variety "Da Mao Hua". Metabolomic analysis showed that terpenoids are the predominant volatiles in both varieties, and the golden flowers are the organs with the highest accumulation of volatile metabolites in both varieties. Comparative profiling of golden flowers identified 131 differentially accumulated metabolites (DAMs). β-Ocimene and α-farnesene were enriched in "Da Mao Hua", whereas α-terpinene and perillene were enriched in "Jiu Feng 2". These DAMs were mainly enriched in terpenoid-related secondary metabolic pathways, including monoterpenoid biosynthesis and sesquiterpenoid/triterpenoid biosynthesis. Transcriptome sequencing generated 268.61 Gb of high-quality data and identified 34 281 differentially expressed genes (DEGs). In golden flowers, DEGs were significantly enriched in pathways related to terpenoid, aliphatic, and phenylpropanoid metabolism. Integrated analysis revealed that both DAMs and DEGs were significantly co-enriched in terpenoid biosynthetic pathways. Quantitative real-time PCR further confirmed that two terpene synthase (TPS) genes involved in β-ocimene and α-farnesene formation, LjTPS15 and LjTPS16, showed significantly lower expression in "Jiu Feng 2", consistent with the reduced accumulation of these volatiles. Collectively, this study reveals the major differences in aromatic compound accumulation and differentially expressed genes between two L. japonica varieties, laying the foundation for subsequent in-depth analysis of their molecular mechanism and providing a theoretical basis for breeding of L. japonica.

Recombinant human arginase 1 (rhArg1) has significant therapeutic potential for the treatment of arginase 1 deficiency (ARG1-D) and arginine-auxotrophic cancers. However, its relatively low catalytic efficiency and limited in vivo stability restrict its clinical application. To enhance the catalytic performance of rhArg1, we employed the protein language model SaProt to perform zero-shot global mutational effect prediction across the entire rhArg1 sequence. Based on the predicted scores, the top 37 candidate variants were selected for experimental validation. Enzyme activity assays revealed that 16 of the 37 variants exhibited higher activity than the wild-type (WT) enzyme. Among them, the V30A, C45F, A56V, and H312P variants showed the most pronounced improvements, with catalytic activities increased by 33%, 23%, 22%, and 90%, respectively, compared with WT. Further kinetic characterization demonstrated that the H312P variant increased by 131% in catalytic efficiency (k_cat/K_M) relative to WT. These results indicate that the protein language model SaProt can effectively guide the rational engineering of arginase. The high-activity variants identified in this study provide promising candidate molecules for the treatment of ARG1-D and arginine-auxotrophic cancers.

Centipeda minima (L.) A. Braun et Aschers. is a traditional medicinal plant. Sesquiterpenoids are its core bioactive components, and sesquiterpene synthases (STSs) are the key enzymes in sesquiterpene biosynthesis. In this study, using C. minima as material, nine full-length candidate CmTPSs genes were screened from published transcriptome data by local Blast alignment combined with phylogenetic analysis. Bioinformatics approaches were used to analyze their sequence features, phylogenetic relationships, and conserved motifs. TBtools was employed to construct gene expression heatmaps for investigating their tissue-specific expression patterns. The catalytic functions of CmTPS1, CmTPS3, CmTPS4, and CmTPS5 were further verified by gene cloning, heterologous expression in Saccharomyces cerevisiae, and GC-MS analysis. Results showed that the nine CmTPSs genes fell into five TPS subfamilies, namely TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g, and displayed distinct tissue-specific expression in roots, stems, leaves, and flowers. CmTPS1 and CmTPS4 were specific caryophyllene synthases, while CmTPS3 and CmTPS5 were multifunctional synthases. This study provides key gene resources for elucidating the molecular mechanism of sesquiterpene biosynthesis in C. minima, and offers theoretical and technical support for the efficient production of its bioactive constituents and the development of its medicinal resources.

The dried roots of Phytolacca acinosa Roxb. are widely utilized in traditional Chinese medicine for their potent diuretic, anti-edematous, and detoxifying properties. Esculentosides, the primary bioactive constituents of P. acinosa, exhibit various pharmacological activities, including anti-inflammatory, anti-tumor, and antifungal effects. However, their clinical potential is hampered by low natural abundance and the inherent challenges of chemical synthesis due to their complex structural scaffolds, while synthetic biology represents a promising strategy for the sustainable and green production of esculentoside. However, the biosynthetic pathways remain unclear, and their key functional enzymes have not been reported yet. Herein, through activity-guided enzyme mining and transcriptome analyses, three novel glycosyltransferases (PaUGT2, PaUGT15, and PaUGT25) were functionally characterized from P. acinosa callus. These enzymes were capable of catalyzing the regio-selective glycosylation of 28-COOH of phytolaccagenin. Notably, PaUGT2 could efficiently catalyze 28-O-glycosylation of esculentoside A and esculentoside B, and PaUGT15 and PaUGT25 also exhibited catalytic activities toward these substrates. These findings not only preliminarily clarified the crucial enzymes for 28-O-glycosylation and elucidated the molecular basis for the structural diversity of oleanane-type esculentosides in Phytolacca plants, but also paved the way for further deciphering the biosynthetic pathway and constructing an artificial pathway of these glycosides.