Proteolysis-targeting chimeras (PROTACs) leverage their "event-driven" mechanism to target traditionally undruggable proteins inaccessible to conventional small molecule inhibitors, thereby significantly expanding the therapeutic landscape of drug discovery. Furthermore, PROTACs overcome the resistance associated with small molecule inhibitors by achieving complete degradation of target proteins rather than transient functional inhibition. However, their sustained catalytic activity may induce off-target effects and systemic toxicity. To mitigate these limitations, stimuli-responsive PROTAC prodrug systems have emerged as a promising strategy to spatiotemporally control PROTAC activation within tumor microenvironments. PROTAC prodrug systems demonstrate tumor microenvironment-responsive activation through endogenous/exogenous stimuli (e.g., light, enzymes, reactive oxygen species), enabling spatiotemporal control of PROTAC liberation. This precision engineering strategy significantly reduces off-target toxicity while maintaining potent on-target protein degradation efficacy. This review provides a systematic overview of cutting-edge advances in stimuli-activatable PROTAC platforms, critically examines current limitations and challenges in the field, and proposes strategic directions for future development to accelerate the evolution of precision cancer therapeutics.

Targeted protein degradation (TPD) technology achieves selective degradation of target proteins through the ubiquitin-proteasome system (UPS) or lysosomal pathways. Among TPD strategies, proteolysis-targeting chimeras (PROTACs) have emerged as a transformative approach in innovative drug discovery, offering unique advantages such as the ability to target "undruggable" proteins, catalytic degradation kinetics, potential to circumvent drug resistance, and high selectivity. However, conventional small-molecule PROTACs depend on ligands binding to target proteins, limiting their effectiveness against molecules without well-defined binding sites, such as transcription factors (TFs) and RNA-binding proteins (RBPs). To overcome this limitation, oligonucleotide-based PROTAC technology has been developed, utilizing sequence-specific nucleic acid recognition to significantly expand the repertoire of degradable targets. This review systematically summarizes recent advances in oligonucleotide-PROTACs, including innovative designs like TF-PROTACs, RNA-PROTACs, G4-PROTACs, and aptamer-PROTACs, along with their therapeutic potentials. We further discuss persisting challenges such as molecular stability and delivery efficiency, and propose future directions for the field. By integrating nucleic acid molecular biology with protein degradation technologies, this interdisciplinary paradigm promises to unlock novel therapeutic strategies for "undruggable" targets and previously intractable diseases.

Breast cancer remains one of the most prevalent malignancies worldwide, with approximately 2.3 million new cases and 670 000 deaths reported globally in 2022. Current therapeutic challenges include the difficulty in targeting breast cancer-associated proteins lacking active sites through traditional small-molecule inhibitors, as well as the high incidence of acquired drug resistance, which collectively undermine treatment outcomes. The advent of proteolysis-targeting chimera (PROTAC) technology has provided a solution, offering a unique protein degradation mechanism to circumvent these limitations. Nevertheless, clinical translation of PROTACs requires addressing inherent limitations such as poor solubility, low bioavailability, and insufficient targeting specificity. To overcome these barriers, multifunctional nano-delivery systems have emerged as a promising technological platform to enhance the therapeutic potential of PROTACs. This review systematically examines the latest advances in nanotechnology-enabled PROTAC delivery for breast cancer treatment, with particular emphasizes rational nanocarrier design principles and synergistic mechanisms of antitumor combination therapies. We further critically analyze persistent translational challenges and propose forward-looking solutions. The integration of nanotechnology with PROTACs not only overcomes the limitations of traditional therapies but also offers new insights into the precise treatment of breast cancer.

Targeted protein degradation (TPD) drugs, which specifically degrade protein of interest, provide novel therapeutic strategies for a spectrum of diseases. However, traditional small-molecule degraders still face challenges such as poor bioavailability, limited availability of E3 ligases, and significant off-target effects. Macromolecule-based TPD agents, characterized by greater design flexibility and enhanced specificity, hold promise for overcoming the limitations of conventional proteolysis-targeting chimeras (PROTACs). This article focuses on macromolecular degradation systems, systematically analyzing and comparing the molecular mechanisms, clinical translation potential, and inherent challenges of different technological approaches based on two major intracellular degradation pathways—the ubiquitin-proteasome system and the lysosomal pathway. Furthermore, we discussed emerging directions, including transmembrane protein degradation and light-controlled protein degradation, offering insights for developing precision-targeted protein degradation-based therapeutic frameworks.

The development of therapeutic interventions for central nervous system (CNS) diseases poses significant challenges. These challenges are primarily attributable to the limitations imposed by the blood-brain barrier (BBB) permeability and the inherent complexity of protein aggregates. Targeted protein degradation (TPD) technology selectively removes pathogenic proteins through the ubiquitin-proteasome system or lysosomal pathways, offering a new strategy for CNS treatment. In comparison with conventional small molecule drugs, TPD employs an event-driven mode of action, capable of degrading β-amyloid and other "undruggable" targets, and demonstrating efficacy at sub-chemometric doses. This is anticipated to markedly reduce the complexity of CNS delivery. The disconnecting of pharmacodynamics and pharmacokinetics further reduces the risk of off-targeting, making it particularly suitable for the CNS, a therapeutic area that requires a high degree of drug safety. Three CNS degraders have already entered clinical trials. This review comprehensively elaborates on the application progress of TPD technologies based on different degradation systems in the treatment of CNS diseases, systematically analyzes the advantages and limitations of each technology, and discusses the common challenges and general optimization strategies for TPD application in CNS disease therapy. The findings aim to provide new insights for the in-depth application of TPD technology in the treatment of CNS diseases.

Neurodegenerative diseases (NDDs) are marked by disruptions in protein homeostasis, including abnormal synthesis, misfolding, aggregation, and defective degradation of pathogenic proteins, leading to neurotoxicity. In recent years, multilayered therapeutic strategies targeting protein homeostasis have been developed, such as transcriptional and translational regulation, conformational stabilization, aggregation inhibition, and protein clearance approaches represented by targeted protein degradation (TPD). Since the pathological lesions of NDDs are primarily located in the central nervous system, effective interventions require that therapeutic agents cross the blood-brain barrier (BBB) and be precisely delivered to diseased brain regions. This review systematically summarizes the pathogenic mechanisms underlying protein homeostasis imbalance in NDDs and highlights current therapeutic strategies, with an emphasis on recent advances and therapeutic potential of TPD approaches. We further overview various brain-targeted nanodelivery systems, including those responsive to the brain microenvironment and capable of targeting specific cells and subcellular organelles, which collectively enable precise trans-BBB delivery and intracellular targeting. These advances support the development of precision therapies aimed at restoring protein homeostasis. This review provides insights into the progress of brain-targeted TPD technologies and their supporting nanodelivery strategies.

Molecular glues can modulate biological functions by specifically inducing interactions between target proteins and E3 ubiquitin ligases, demonstrating significant advantages for disease treatment. However, the discovery of most molecular glues still relies on serendipity, lacking systematic methods and rational design strategies. In recent years, rapid advances in chemical biology and in-depth research on molecular glue-protein interaction mechanisms have gradually enabled systematic development of molecular glues. Therefore, this review aims to organize discovery strategies and representative molecules from three perspectives: serendipitous discovery, screening methods, and rational design, providing critical insights and references for future molecular glue development.

Proteostasis is a core biological process through which cells maintain the stability and functional integrity of the proteome via a multi-level quality control network. As a key molecule in the ubiquitination regulatory pathway, the deubiquitinase USP7 can drive tumorigenesis and progression by altering the ubiquitination status of substrate proteins when its expression is aberrant or activity is dysregulated, thus emerging as a critical target in precision cancer therapy. This article systematically outlines the structural features of USP7, its role in regulating substrate protein homeostasis, and its biological functions in tumor development. It focuses on recent advances in USP7-based anti-tumor strategies, including the development of small-molecule inhibitors, applications of proteolysis-targeting chimeras (PROTACs) technology, and exploration of novel approaches such as deubiquitinase-targeting chimeras (DUBTACs). This review provides a systematic theoretical framework and cutting-edge research directions for the development of USP7-targeted anti-tumor drugs.

Traditional Chinese medicines (TCMs), with its rich historical legacy and demonstrated efficacy in treating chronic non-communicable diseases, particularly in addressing cancer, cardiovascular diseases, diabetes, and chronic respiratory diseases, presents unique challenges for scientific investigation due to its complex chemical composition and poorly understood mechanisms of action. This review systematically examines the evolution of target fishing technology, evaluates established target identification technology and ligand fishing technology in TCMs research, and synthesizes recent advancements in applying these methodologies to elucidate the therapeutic mechanisms of TCMs for major chronic diseases, with special emphasis on component-target interactions. By providing a critical analysis of current applications and future directions, this work aims to advance the integration of target fishing technologies into modern TCMs research and establish a robust framework for mechanistic studies in this field.

Alzheimer's disease (AD) is a complex neurodegenerative disorder, and its pathological mechanisms are not yet fully understood. In recent years, mitochondrial dysfunction has been recognized as one of the important pathological features of AD. As the central hub of intracellular energy metabolism, mitochondria maintain ATP homeostasis through the tricarboxylic acid cycle and oxidative phosphorylation under physiological conditions, and they are involved in important physiological processes. However, under the pathological state of AD, mitochondrial function undergoes statistically significant changes, characterized by bioenergetic metabolism imbalance, impaired mitophagy, abnormal dynamics (including excessive fission and fusion defects), and excessive production of mitochondrial reactive oxygen species. Moreover, abnormal interactions between mitochondria and the endoplasmic reticulum disrupt cellular homeostasis, thereby significantly accelerating neuronal degeneration by exacerbating the pathological processes of AD. This article systematically reviews the core pathological features of AD, focusing on the physiological functions of mitochondria and their dynamic changes during the pathological progression of AD. Finally, this article summarizes the anti-AD therapeutic strategies that regulate mitochondrial function, including small molecule drugs and natural products, providing new ideas and potential solutions for the treatment of AD.

Neuroinflammation plays a pivotal regulatory role in the onset and progression of various central nervous system diseases. As a key immune sensor, the NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome, upon activation, can exacerbate neuroinflammation, leading to neuronal injury and death, thereby accelerating the pathological processes of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Consequently, the NLRP3 inflammasome has emerged as an important therapeutic target for modulating neuroinflammation. In recent years, significant progress has been made in the development of NLRP3 inflammasome inhibitors. Numerous novel inhibitors have been identified that effectively reduce inflammatory responses and protect neurons from damage, demonstrating promising efficacy in animal models, particularly in improving cognitive and motor functions. This review will summarize the canonical activation mechanisms of the NLRP3 inflammasome, explore its pathogenic roles in central nervous system diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, and depression, and provide an overview of recent advances in NLRP3-targeted drug development, aiming to offer new insights and strategies for the treatment of central nervous system diseases.

Natural components are crucial sources for drug discovery. Based on the central dogma of molecular biology, we proposed a novel paradigm for classifying natural products by categorizing natural components into "direct gene-encoded components" (including nucleic acids and peptides) and "indirect gene-encoded components" (encompassing primary metabolites and secondary metabolites). Utilizing multi-omics and artificial intelligence technologies, we systematically analyzed the nuclear and organellar genomes of 1 037 medicinal species sourced from eight authoritative global pharmacopeias, integrating multidimensional data resources to establish the gene-encoded natural diversity components repository (GNDC). This paper comprehensively describes: (1) The construction methodology of GNDC, including data integration strategies, standardized annotation pipelines, and rigorous quality control; (2) Core features and data scale: GNDC is currently the world's largest repository of medicinal natural products, housing over 234 million gene-encoded (directly or indirectly) natural components. It encompassed four specialized sub-databases: HerbalMDB for 2.32 million secondary metabolites, HerbalPDB for 229 million small peptides, HerbalRDB for 2.38 million small RNAs, and HerbalCDB for 0.26 million carbohydrates; (3) Exploring the application prospects of GNDC in the modernization of traditional medicine research. GNDC will provide an unprecedented expansive "chemical space" for drug discovery. It will powerfully drive a paradigm shift in drug development from "experience-oriented" to "big data-driven" approaches, offering a transformative framework for traditional medicine research.

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease closely associated with metabolic syndrome, and its animal models are essential for investigating the pathological mechanisms and disease progression of NAFLD. Currently, the most commonly used NAFLD animal models include the high-fat, methionine- and choline-deficient diet (HFMCD) and the high-fat diet (HFD)-induced mouse models. In this study, we compared NAFLD mouse models established by an 8-week HFMCD diet and a 12-week HFD, systematically evaluating differences in liver weight, histological alterations, lipid metabolism, and inflammation-related gene expression. The experimental results showed that, compared with the normal chow diet (NCD) group, mice in the HFMCD group exhibited significant reductions in body weight and blood glucose levels, whereas those in the HFD group exhibited significant increases. Both dietary models led to an increase in liver weight; however, hematoxylin and eosin (H&E) staining revealed more severe liver damage in the HFMCD group. Oil red O staining indicated a significantly greater degree of hepatic lipid accumulation in the HFMCD group than in the HFD group. Quantitative PCR (qPCR) analysis demonstrated that, compared with the NCD group, both the HFMCD and HFD groups exhibited significantly elevated hepatic expression of inflammatory cytokines. Sirius red staining demonstrated pronounced hepatic fibrosis in the HFMCD group, accompanied by a significant upregulation of genes associated with collagen synthesis, whereas the HFD group exhibited no evident fibrotic alterations. F4/80 immunohistochemical staining showed increased hepatic macrophage infiltration in both models, with a significantly higher number of F4/80-positive macrophages in the HFMCD group than in the HFD group. In conclusion, the HFMCD and HFD-induced NAFLD models exhibit distinct pathological characteristics and molecular mechanisms. The HFMCD model is more suitable for studying the progression of NAFLD to non-alcoholic steatohepatitis (NASH), whereas the HFD model is more appropriate for investigating the metabolic mechanisms of simple steatosis. This study provides experimental evidence for NAFLD research and offers guidance for selecting appropriate animal models based on specific research objectives. All experimental animals used in this study were approved by the Animal Care and Use Committee of Nanjing University and were handled in strict accordance with the guidelines and recommendations provided by the Nanjing University's Animal Ethics Committee.

Breast cancer bone metastasis disrupts bone metabolism, enhancing osteogenic differentiation can improve the metastatic microenvironment and treatment outcomes. Thus, developing agents that inhibit breast cancer and promote osteogenesis may offer new strategies. Tripterygium wilfordii possesses antitumor and bone-regulating properties, but its toxicity limits clinical application. Here, a carbon dot was synthesized from Tripterygium wilfordii (Tripterygium wilfordii carbon dots, T-CDs) and characterized by spectral analysis, dynamic light scattering, and transmission electron microscopy. T-CDs exhibited a near-spherical morphology with a lattice spacing of 0.20 nm, an average size of 5.38 nm, and a ζ potential of -8.46 mV. The surface was rich in -OH, C=O, and C=C groups, with ultraviolet absorption at 280 nm. T-CDs disrupted redox homeostasis in breast cancer cells, inducing ferroptosis and immunogenic cell death. Moreover, T-CDs modulated cytokine secretion and promoted osteogenic differentiation, offering potential for the treatment of breast cancer and its bone metastasis.

To explore the mechanism of Lvjiaobuxue Granule (LBG) in improving postpartum Qi and blood deficiency in rats through milk metabolomics combined with biological target network analysis, the model of postpartum Qi and blood deficiency in rats was established by "orbital bloodletting combined with exhaustive swimming under negative gravity". The behavioral indexes, blood routine parameters, immune organ index and changes of immunoglobulin G (IgG) in milk of rats were detected to evaluate the improvement effect of LBG on postpartum Qi and blood deficiency in rats. LC-MS milk metabolomics was applied to find the differential metabolites in the milk of rats with postpartum Qi and blood deficiency model regulated by LBG. Combined with biological target network analysis, the material basis of the efficacy, action targets and signaling pathways of LBG in improving postpartum Qi and blood deficiency were predicted, and molecular docking was used for verification. The animal experiments in this study were conducted in compliance with the regulations of the Scientific Research Ethics Committee of Shanxi University and approved by the Animal Experimentation Ethics Review (approval No.: SXULL2023022). LBG significantly improved the abnormal behavior, decreased blood routine parameters, immune organ damage and decreased IgG in milk of rats with postpartum Qi and blood deficiency model (P < 0.05, P < 0.01). Twenty endogenous metabolites were found to be differential in milk, serving as potential biomarkers for LBG to improve postpartum Qi and blood deficiency in rats, and these metabolites mainly included amino acids, fatty acids and nucleotides, etc. Biological target network analysis showed that its potential mechanism might be related to the signaling pathway of steroid hormone biosynthesis. The key components in LBG, luteolin, caffeic acid and ferulic acid, might respectively regulate the core targets of CYP19A1, CYP1A1, CYP1A2 and CYP3A4, affect the changes of linoleic acid endogenous metabolites in milk, and regulate the biosynthesis of steroid hormones to play its role in improving postpartum Qi and blood deficiency. In addition, the results of molecular docking showed that the three key components in LBG had significant binding effects with their core targets. LBG may affect the linoleic acid level in milk by regulating CYP19A1, CYP1A1, CYP1A2 and CYP3A4 with luteolin, ferulic acid and caffeic acid respectively, and then regulate the signaling pathway of steroid hormone biosynthesis to improve postpartum Qi and blood deficiency in rats.

Right ventricular (RV) dysfunction is a pivotal prognostic determinant in patients with pulmonary arterial hypertension (PAH). Hypoxic stimulation induces hypoxic pulmonary vasoconstriction (HPV) and exerts cardiotoxic effects, thereby accelerating RV remodeling in PAH. Previous in vitro vascular ring experiments demonstrated that α-phellandrene (PE) exhibits vasodilatory effects, but its impact on hypoxia-induced RV structural and functional alterations and underlying mechanisms remain unclear. To address this, we established a hypoxia-induced pulmonary arterial hypertension (HPH) rats model and comprehensively evaluated the therapeutic effects of PE on multi-scale RV pathological remodeling through echocardiography, histomorphometric analyses, and Western blot. Additionally, the molecular mechanisms underlying PE-mediated modulation of the nitric oxide (NO)-soluble guanylyl cyclase (sGC)-cyclic guanosine monophosphate (cGMP) signaling pathway were investigated using H9C2 cardiomyocytes. Results demonstrated that compared with the model group, PE treatment significantly improved RV systolic function in HPH rats, as evidenced by increased tricuspid annular plane systolic excursion, pulmonary artery acceleration time, fractional area change, and RV ejection fraction. Concurrently, PE reduced RV free wall thickness in diastole and RV inner diameter, effectively attenuating hypoxia-induced RV remodeling. PE administration also markedly decreased mean pulmonary artery pressure and RV hypertrophy index compared to the model group. Histomorphological analysis demonstrated that PE treatment significantly reduced collagen deposition in RV tissues and downregulated expression of fibrotic markers, including α-smooth muscle actin, collagen Ⅰ and collagen Ⅲ. Mechanistic investigations revealed that PE upregulated endothelial nitric oxide synthase expression in both rats RV tissues and H9C2 cardiomyocytes, enhanced NO bioavailability, and activated sGC- cGMP signaling axis. These molecular effects collectively ameliorated hypoxia-induced oxidative damage and attenuated fibrotic progression. This study reveals that PE ameliorates hypoxia-induced RV remodeling and dysfunction by modulating the NO-sGC-cGMP signaling pathway to reduce oxidative damage and fibrotic remodeling in hypoxic conditions. These findings provide experimental evidence for developing RV-targeted therapeutic strategies against HPH. All animal experiments were approved by the Institutional Animal Care and Use Committee of Qinghai University (approval No: PJ-202302-01).

Terpinen-4-ol (T4O) is a bioactive monoterpenoid with significant cardiovascular pharmacological activity. This study investigated T4O's protective effects against high glucose (HG) induced injury in rat cardiomyocytes (H9C2) and its impact on silencing the sirtuin 1 (SIRT1)/peroxisome proliferator-activated receptor-γ (PPARγ) signaling pathway. In the experiment, H9C2 cells were pre-incubated with different doses of T4O for 2 h, followed by co-incubation with HG for 48 h to create an injury model. Cell survival was determined using the MTT assay. Intracellular reactive oxygen species (ROS) levels were detected using the fluorescent probe DCFH-DA. Apoptosis rates were detected using flow cytometry. Mitochondrial membrane potentials were observed using JC-1 staining. Western blot was used to detect the expression levels of the apoptosis-related proteins Bcl-2, Bax, Caspase-9, and Caspase-3, as well as SIRT1 and PPARγ. Immunofluorescence was used to observe the localization and expression of SIRT1 and PPARγ. Silencing SIRT1 expression in H9C2 cells was achieved by SIRT1 siRNA transfection. The effects of T4O on ROS content, apoptosis rate, mitochondrial membrane potential, and expression of the aforementioned proteins in HG-induced H9C2 cells were then observed. Compared to the HG group, T4O significantly improved HG-induced injury to H9C2 cells. It decreased ROS levels and the apoptosis rate, increased the mitochondrial membrane potential, and down-regulated the expression levels of the Caspase-9 and Caspase-3 proteins induced by HG. Meanwhile, it increased the Bcl-2/Bax ratio and SIRT1 and PPARγ expression levels (P < 0.05, P < 0.01). After SIRT1 was silenced, compared with the negative control + HG group, the SIRT1 siRNA + HG group exhibited significantly increased intracellular ROS levels and apoptosis rates, reduced mitochondrial membrane potential, and upregulated of Caspase-9 and Caspase-3 protein expression. while the Bcl-2/Bax ratio and expression levels of SIRT1 and PPARγ were further downregulated (P < 0.05, P < 0.01). Co-treatment with SIRT1 silencing and T4O reversed the protective effect of T4O against H9C2 cells damage. In conclusion, T4O ameliorates HG-induced injury to H9C2 cells by activating the SIRT1/PPARγ signaling pathway. This provides a new idea and potential target for developing anti-myocardial injury drugs.

Urtica fissa, a commonly used traditional Chinese herb, frequently causes contact dermatitis during application. However, the inflammatory components have not been reported. In this study, RAW264.7 cell model was employed first to identify the dichloromethane fraction as the primary inflammatory fraction. Then, using chromatographic techniques such as silica gel column chromatography and preparative HPLC, combined with HR-MS, NMR, and UPLC-IM-Q-TOF-MS analysis, 44 compounds, including nettlol, cinnamic acid, and salicylic acid, were identified from the dichloromethane fraction. Subsequently, the potential inflammatory components were confirmed by transcriptomic analysis and molecular docking. Finally, the inflammatory effects of these candidates were evaluated by an open topical application test on mouse skin. The results showed that cinnamic acid and salicylic acid significantly induced skin erythema and ear swelling in mice, and markedly elevated histamine and NO levels in serum and skin, confirming them as the key inflammatory constituents. All animal experiments were reviewed and approved by the Ethics Committee of Shanghai Jiao Tong University Laboratory Center (A2025060-001). The findings of this study provided a scientific basis for further research and application of U. fissa.

This study employed CHO-K1-FSHR-CRE-Luc transgenic cells as target cells to detect the biological activity of rhFSH-CTP using a luciferase detection system. Key parameters of the method were optimized, and its performance was validated according to the requirements of the 2020 Edition of Pharmacopoeia of the People's Republic of China (Vol IV), including relative accuracy, intermediate precision, linearity and range, and reliability results. The results showed that rhFSH-CTP conformed to the four-parameter equation: y = (A-D)/[1+(x/C)^B]+D, exhibiting an excellent dose-effect curve. The geometric means of relative biological activities (n = 8) at five titer levels (50%, 80%, 100%, 125%, and 200%) were (53.87 ± 2.47)%, (81.58 ± 1.66)%, (96.76 ± 4.64)%, (127.03 ± 8.19)%, and (207.62 ± 15.52)%, respectively. The relative bias (RB) and its 90% confidence intervals were 7.7% (4.2%, 10.8%), 2.0% (0.6%, 3.3%), -3.2% (-6.4%, -0.3%), 1.6% (-2.9%, 5.8%), and 3.8% (-1.6%, 8.9%), with absolute values of RB and confidence intervals all less than 11%. The upper limits of geometric coefficients of variation (GCV) and their confidence intervals for five titer levels were less than 20%. The linear range spanned 50%-200%, with a slope of 0.974 2 and R² = 0.994 8, meeting the criteria of General Chapter 9401 in Volume IV of the 2020 Pharmacopoeia. Additionally, the deviation parallelism and regression terms satisfied the reliability test requirements of General Chapter 1431 (biological assay statistics) in the same pharmacopoeia edition. In conclusion, this study successfully established a method for detecting the biological activity of rhFSH-CTP, which demonstrates good validity and reliability.

Baicalin eye drops have broad clinical application prospects in the treatment of ocular diseases. However, there is still a lack of qualitative and quantitative research on the related substances in them. This study first established an analytical method for related substances in baicalin eye drops, providing a scientific basis for product development and quality control. Through LC-Q/TOF-MS/MS, seven related substances were preliminarily identified in baicalin eye drops, namely scutellarin, oroxylin A-7-O-glucuronide, oroxin A, quercetin-7-O-glucuronide/quercetin 3-O-glucuronide, chrysin-7-O-glucuronide, baicalein, and scutellarein. The quantitative analysis of oroxin A, scutellarin and oroxylin A-7-O-glucuronide was carried out by HPLC. The chromatographic column was the NanoChrom AR C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase was 0.1% phosphoric acid aqueous solution (A)-acetonitrile (B), with gradient elution. The column temperature was 20 ℃, the flow rate was 0.8 mL·min^-1, and the detection wavelength was 254 nm. The established method has strong specificity, high sensitivity, and good precision. In this study, a method for the qualitative and quantitative analysis of related substances in baicalin eye drops was established, providing a basis for ensuring their quality.

An UPLC-QTRAP-MS/MS analytical method for six blood components (rhein, aloeemodin, sinapic acid, liquiritin, isoliquiritin apioside, and wogonoside) in Daxiefei Decoction was established and applied to the pharmacokinetic study in normal and chronic obstructive pulmonary disease (COPD) model rats to reveal their absorption kinetics characteristics and elimination rules in vivo, in order to provide a scientific basis for the clinical application and further development of Daxiefei Decoction. The plasma concentrations of each substance at different time points were determined by ultra-performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry (UPLC-QTRAP-MS/MS), and the pharmacokinetic parameters of each component were calculated by pharmacokinetic software DAS 2.0. The results showed that the linear relationship between the six components was good in their respective ranges, and the methodological investigation met the requirements. The pharmacokinetic parameters of six components, such as peak concentration (C_max), peak time (t_max), half-life (t_1/2), area under the plasma concentration-time curve (AUC_0-t), were successfully calculated by using the pharmacokinetic software DAS 2.0 non-compartmental model. There were significant differences in the pharmacokinetic characteristics of the active ingredient group between the normal group and the model group, suggesting that the body was in different physiological states, and the change of the body state would have a certain impact on the absorption and elimination of the active ingredient of Daxiefei Decoction. The experiment was approved by the Laboratory Animal Ethics Committee of Nanjing University of Chinese Medicine (No. 202406A085).

A HPLC-QAMS method was established for the simultaneous determination of 15 components in Xiao'er Kechuanling oral liquid, providing a method for its quality control. The analysis was performed on a 30 ℃ Kromasil C18 column (250 mm × 4.6 mm, 5 μm), with the mobile phase comprising of acetonitrile-0.1% formic acid flowing at 1.0 mL·min^-1 in a gradient elution manner. The injection volume was 5 μL and the detection wavelength was set at 254 nm. Chlorogenic acid was used as an internal standard to calculate the relative correction factors of the other fourteen components, after which the content determination was made. Cluster analysis and principal component analysis were conducted on 22 batches of samples. Fifteen components showed good linear relationships within their own ranges (r ≥ 0.999 5), whose average recoveries were 96.52%-99.02% with the RSDs of 0.57%-1.40%. The result obtained by HPLC-QAMS approximated those obtained by external standard method. Cluster analysis of 22 batches of samples was divided into four classes. Four principal components were extracted by principal component analysis, and the contribution rate reached 81.706%. This accurate and specific method can be used for the quality control of Xiao'er Kechuanling oral liquid.

Protoberberine alkaloids exhibit significant antitumor and angiogenesis-regulating activities, yet their specific effects on angiogenesis intervention and metabolic regulatory mechanisms remain systematically elucidated. In this study, ultra-high performance liquid chromatography coupled with quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q Exactive Orbitrap HRMS) was employed to identify the chemical composition profiles of Coptidis Rhizoma and Corydalis yanhusuo. The anti-angiogenic effects of the major protoberberine-type alkaloid components were evaluated using the quail chorioallantoic membrane (qCAM) model and human umbilical vein endothelial cell (HUVEC) proliferation assays, while vascular endothelial growth factor receptor 2 (VEGFR2) mRNA levels were detected via qPCR experiments. Further analysis was conducted using desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) technology to examine the metabolic profile changes induced by the intervention of key anti-angiogenic components. The interaction between these components and critical angiogenesis targets was characterized by integrating network pharmacology with molecular docking technology, followed by a "structure-activity" analysis. A total of 13 alkaloid components were identified in Coptidis Rhizoma, and 17 alkaloid components were identified in Corydalis yanhusuo, with protoberberine-type alkaloids being the predominant type in both. Both the qCAM model and HUVEC experiments demonstrated that oxyberberine exhibited the most significant anti-angiogenic activity. These metabolic abnormalities characterized by suppressed energy and lipid metabolism, may inhibit angiogenesis by affecting cell membrane structure (phosphatidic acid, sphingomyelin, phosphatidylcholine), signal transduction (VEGFR2), oxidative stress, and inflammatory responses. Molecular docking and network pharmacology analyses demonstrated that C-8 oxidation enhances oxyberberine's binding affinity to EGFR and JAK2 through reduced binding energy and increased weak interactions, ultimately conferring superior anti-angiogenic efficacy compared to berberine. These findings systematically elucidate the structure-activity relationship and metabolic basis of protoberberine-type alkaloids in angiogenesis modulation, providing novel insights for anti-angiogenic drug development.

Carbon quantum dots (CDs) are a class of zero-dimensional carbon nanomaterials with remarkable fluorescent properties, consisting of carbon nanoparticles with dimensions below 10 nm. Due to its advantages of excellent optical properties, good water solubility and biocompatibility, it shows great potential in photothermal/photodynamic therapy of cancer. In recent years, carbon quantum dots prepared from traditional Chinese medicine as a carbon source have attracted extensive attention from scientists. In this study, we synthesized carbon quantum dots of five Chinese medicines, Ganoderma lucidum, Poria cocos, Dendrobium officinale, Coix lacryma, and Ginger, in one step by a conventional hydrothermal method, and screened out the Ganoderma lucidum CDs with the best fluorescence quantum yields and photothermal conversion properties. The results show that the Ganoderma lucidum CDs prepared in this study, with a particle size of (6.40 ± 1.31) nm, have good in vivo and in vitro fluorescence imaging properties, demonstrating certain tumor targeting properties. And they can generate photothermal/photodynamic synergy to efficiently kill pancreatic cancer cells under the mediation of near-infrared laser. The animal experiment has been approved by the Animal Ethics Committee of Guizhou University of Chinese Medicine (No. 20230022).

O-Methyltransferases (OMTs) are key enzymes involved in the methylation modification of benzylisoquinoline alkaloids (BIAs), and their functional studies contribute to elucidating the molecular mechanisms underlying BIA biosynthesis. In this study, a novel OMT gene was successfully cloned from Stephania cephalantha and subjected to bioinformatics analysis and in vitro functional characterization. The coding sequence (CDS) of ScOMT26 is 1 092 bp in length, encoding a protein of 363 amino acids with a molecular formula of C₁₈₂₇H₂₈₇₉N₄₆₇O₅₄₃S₁₈, classified as a stable hydrophobic protein. In vitro enzymatic assays demonstrated that ScOMT26 catalyzes the methylation of the C4′-hydroxyl group of coclaurine to produce 4′-methylcoclaurine. The optimal reaction conditions were identified as pH 7.0 in 50 mmol·L^-1 potassium phosphate buffer at 25 ℃. This study represents the first report of cloning and functional characterization of an OMT from S. cephalantha, providing critical insights into the role of OMT genes in the biosynthetic pathway of benzylisoquinoline alkaloids.