Background: Stomach adenocarcinoma (STAD) is a common malignant tumor within the digestive system, characterized by significant morbidity and mortality rates. The identification of innovative biomarkers or therapeutic targets for STAD is of utmost importance. A deeper understanding of the molecular mechanisms underlying STAD progression may facilitate the identification of novel prognostic indicators and therapeutic strategies. This investigation aims to assess the expression patterns of alpha-2-macroglobulin (A2M) across various tumors and their corresponding pathological stages, utilizing data from The Cancer Genome Atlas (TCGA) and University of Alabama at Birmingham Cancer Analysis Portal (UALCAN) databases. Methods: To evaluate the influence of A2M on survival prognosis, we employed the Kaplan-Meier method alongside Cox and receiver operating characteristic (ROC) analysis. Additionally, Tumor Immune Estimation Resource, Version 2 (TIMER2.0) was utilized to examine its impact on the infiltration of immune cells within tumors. By employing R programming, STAD samples were divided into high-expressi on and low-expression groups based on A2M gene expression levels. Differentially expressed genes (DEGs) were subsequently identified, followed by enrichment analyses, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis (GSEA). We further selected STAD cells exhibiting high A2M expression and utilized CRISPR/Cas9 technology to silence A2M, to investigate its effects on cell viability, migration, invasion, and colony formation capabilities. Results: Bioinformatics analysis indicated that A2M is highly expressed in STAD tumor tissues compared to normal gastric tissues. Patients exhibiting elevated A2M levels experienced shorter survival periods compared to those with lower expression levels, with Cox and ROC analyses suggesting A2M's potential as a prognostic biomarker. This implies that A2M plays a role in promoting STAD progression and functions as an oncogene. Pathway enrichment analyses demonstrated that A2M facilitates epithelial-mesenchymal transition (EMT) in STAD cells, showing a significant correlation with EMT marker vimentin and EMTrelated genes. Furthermore, A2M exhibited a positive correlation with the infiltration of various immune cells in STAD tissues, displaying strong associations with multiple immune cell markers. A2M expression also influences the responsiveness of STAD patients to immunotherapy and small-molecule drug therapies. Cell experiments indicated that the silencing of A2M expression led to reduced STAD cell viability, migration, invasion, and colony formation, alongside a decrease in the expression of the mesenchymal marker vimentin. Conclusions: These findings suggest that A2M promotes EMT through the upregulation of vimentin expression, thereby facilitating the malignant progression of STAD. Thus, A2M emerges as a promising therapeutic target that warrants further investigation to refine treatment strategies and improve patient outcomes in STAD.

Abstract Ferroptosis is an iron‐dependent form of regulated cell death driven by lipid peroxidation, characterized by distinct ultrastructural alterations such as mitochondrial shrinkage and disruption of cristae. As an emerging therapeutic target in oncology, ferroptosis opens new therapeutic perspectives for a range of diseases, including cancer. Recent studies have demonstrated that ferroptosis induction exhibits superior efficacy in tumor regulation compared to certain conventional treatment modalities. However, the process of ferroptosis is intricately regulated by multiple signaling pathways involving iron metabolism, lipid metabolism, and disturbances in redox homeostasis, thereby limiting the effectiveness of single‐induction strategies. With advances in nanotechnology, a variety of nanomaterials have been engineered to induce ferroptosis through multi‐level regulatory systems or to serve as delivery vehicles for small‐molecule agents, significantly enhancing targeting capability toward tumor tissues. This review systematically summarizes recent advances in the molecular mechanisms, regulatory networks, pathological roles, small‐molecule inducers, and nanocarrier‐based targeted therapeutic strategies related to ferroptosis. It should be noted that, as a rapidly evolving research field, the application of ferroptosis in cancer therapy remains at an early stage. This article further discusses key challenges and future directions for its translation into clinical precision therapies, and concludes with critical scientific questions that demand urgent investigation in future research.

Differentiation therapy holds significant potential for the treatment of multiple myeloma (MM). We previously identified that the aminopeptidase N (APN) inhibitor Bestatin promotes MM cell differentiation. Herein, we elucidate the underlying molecular mechanisms of this process. Utilizing MM1.S, U266, and RPMI-8226 cell lines, a combination of CCK-8 assays, flow cytometry, Wright-Giemsa staining, Western blotting, qRT-PCR, ELISA, APN enzymatic activity analysis, SA-β-gal staining, and bioinformatic analyses revealed elevated APN expression across all cell types. Bestatin treatment induced MM cell differentiation in a concentration-dependent manner, which was accompanied by the upregulation of the differentiation marker CD49e, increased immunoglobulin light chain secretion, elevated cellular senescence, and a concomitant suppression of cell proliferation and APN enzymatic activity. Mechanistically, Bestatin exerts its effects by downregulating the CD79B/BTK signaling pathway, thereby activating the downstream transcription factor STAT3. Consistent with this axis, direct inhibition of CD79B/BTK alone was sufficient to induce differentiation, while blockade of STAT3 completely abrogated the differentiation-promoting effect of Bestatin. The APN-neutralizing antibody (WM15) yielded consistent results with Bestatin, further validating this regulatory axis. Furthermore, both the CD79B/BTK inhibitor Ibrutinib and the STAT3 agonist GCDA potentiated the cytotoxicity of the clinical MM drug Ixazomib. Bestatin itself synergized with Ixazomib and enhanced the anti-proliferative effect of IL-6. In summary, our findings establish that the APN inhibitor Bestatin induces MM cell differentiation via the CD79B/BTK-STAT3 signaling axis. Targeting this pathway represents a promising strategy to enhance the efficacy of Ixazomib, providing a compelling rationale for novel combination therapies in MM.

Chemoresistance, inadequate tumor targeting, and severe systemic toxicity remain key barriers to effective breast cancer treatment. Ferroptosis-inducing strategies have emerged as a promising therapeutic avenue, yet their efficacy is frequently compromised by the intrinsic antioxidant defense system and insufficient immune activation within the tumor microenvironment (TME). Herein, a hyaluronic acid (HA)-functionalized, berbamine (BBM)-loaded iron-based metal-organic framework (MOF) nanoplatform (HA-FeTCPP@BBM, HAFTB) was engineered. This nanosystem executes a "three-in-one" synergistic strategy that coordinates FeTCPP-mediated sonodynamic therapy (SDT), iron-dependent chemodynamic therapy (CDT), and BBM-mediated chemotherapy and ferroptosis sensitization, while HA functionalization facilitates tumor-oriented delivery to enhance the induction of ferroptosis-associated immunogenic cell death in triple-negative breast cancer (TNBC). Upon ultrasound (US) exposure, the HAFTB scaffold functions simultaneously as a sonosensitizer and an iron source. Ultrasound activation initiates sonodynamic reactions and enhances Fenton-like catalytic activity, substantially boosting reactive oxygen species (ROS) generation. Moreover, the localized release of BBM contributes to glutathione peroxidase 4 (GPX4) suppression and weakens the cellular antioxidant defense system, thereby sensitizing tumor cells to ferroptotic stress. This tripartite oxidative stress results in extensive lipid peroxidation and robust ferroptotic cell death. Abundant damage-associated molecular patterns (DAMPs) are concurrently released, supporting the occurrence of ferroptosis-associated immunogenic responses and subsequent immune activation. In an orthotopic TNBC mouse model, HAFTB integrated with SDT achieves pronounced tumor suppression with low systemic toxicity, while transcriptomic profiling supports the involvement of ferroptosis- and oxidative stress-related pathways. Overall, the principal advance of this cascade nanoplatform lies in the coordinated integration of FeTCPP-mediated SDT, iron-dependent CDT, and BBM-associated ferroptosis sensitization within an HA-assisted MOF delivery system, providing a promising therapeutic strategy for TNBC.

NUC1031 is a gemcitabine ProTide prodrug which is currently undergoing phase III. CD13 inhibitor bestatin is utilized as an adjunct therapy in conjunction with chemotherapy for cancer treatment, which has limitations in cytotoxic efficacy. In this study, we designed and synthesized a novel series of bestatin-gemcitabine's ProTide prodrug conjugates aimed at enhancing the antitumor efficacy of NUC1031. The representative compound 5f demonstrates a 10-fold increase in anti-proliferative activity compared to NUC-1031, with an IC50 of 8.5 nM against the prostate cancer cell line 22Rv1. In vitro and in vivo pharmacokinetic studies revealed that compound 5f gradually degrades into the metabolic product 17, potentially extending its anti-tumor activity. 5f demonstrates significant in vivo anti-tumor activity in 22Rv1 xenograft tumor models. Our findings indicate that 5f shows strong potential for further development as a candidate for the treatment of prostate cancer.

The CD13 inhibitor ubenimex is used as an adjuvant drug with chemotherapy for the treatment of cancer due to its function as an immunoenhancer, but it has limitations in its cytotoxic efficacy. The proteasome inhibitor ixazomib is a landmark drug in the treatment of multiple myeloma with a high anti-cancer activity. Herein, we conjugated the pharmacophore of ubenimex and the boric acid of ixazomib to obtain a dual CD13 and proteasome inhibitor 7 (BC-05). BC-05 exhibited potent inhibitory activity on both human CD13 (IC50 = 0.13 μM) and the 20S proteasome (IC50 = 1.39 μM). Although BC-05 displayed lower anti-proliferative activity than that of ixazomib in vitro, an advantage was established in the in vivo anti-cancer efficacy and prolongation of survival time, which may be due to its anti-metastatic and immune-stimulating activity. A pharmacokinetic study revealed that BC-05 is a potentially orally active agent with an F% value of 24.9%. Moreover, BC-05 showed more favorable safety profiles than those of ixazomib in preliminary toxicity studies. Overall, the results indicate that BC-05 is a promising drug candidate for the treatment of multiple myeloma.

目的 探究血管紧张素受体AT1相关受体蛋白(puta-tive receptor protein related to AT1,APJ)同源二聚体对人脐静脉内皮细胞(HUVECs)行为,即增殖、迁移和成管的作用.方法 将HUVECs随机分为对照组(PBS)、APJ单体组[爱帕琳肽13(Apelin-13)+TM1]和APJ同源二聚体组(Apelin-13+PBS),分别用Western blot和基质辅助激光解吸飞行时间质谱仪(MALDI-TOF MS)检测APJ和APJ同源二聚体在HUVECs中的表达;实时细胞分析仪(RTCA)检测Apelin-13引起50％最大效应的浓度(EC50);CCK-8法测定HUVECs的细胞活力;划痕实验检测HUVECs的迁移能力;人工基底膜(matrigel)检测HUVECs的成管.结果 Western blot和MALDI-TOF MS实验结果显示,APJ和APJ同源二聚体均表达于HUVECs中.Apelin-13的EC50为2.26×10-8 mol·L-1,且引起最大效应的浓度为1.0×10-6 mol·L-1.CCK-8实验、迁移实验和成管实验结果显示,各组细胞随着时间的延长逐渐向划痕裸区迁移,镜下也观察到各组细胞均出现成管,数据统计发现Apelin-13+PBS和Apelin-13+TM1组的HUVECs增殖、迁移和成管能力均较PBS组明显增加,且Apelin-13+PBS组的增殖、迁移和成管能力明显好于Apelin-13+TM1组(P<0.05).结论 APJ同源二聚体对HUVECs的增殖、迁移和成管均具有促进作用,且效果优于APJ单体.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) has emerged as a promising therapeutic target for the treatment of hyperlipidemia. In discovery of novel small molecules that interfere PCSK9/LDLR protein-protein interaction (PPI), structural modification was performed based on our previously derived compounds. A series of [5,5'-bibenzo[d][1,3]dioxol]-6-amine analogs were designed and synthesized for the activity evaluation. In the PCSK9/LDLR PPI impairing test, molecules D28 and D29, exhibited remarkable inhibitory potency with IC50 values of 8.30 and 6.70 μM compared with SBC-115337 (17.89 μM), respectively. Molecular docking predicted the binding pattern of compounds D28 and D29 in the LDLR binding site of PCSK9. Hydrophobic interactions play an important role in the binding of aromatic molecular fragments to the pockets in the PCSK9/LDLR binding interface. Further LDLR expression and LDL uptake studies revealed that both D28 and D29 restored LDLR expression on the surface of hepatic HepG2 cells and improved extracellular LDL uptake in the presence of PCSK9. It is significant that molecules D28 and D29 exhibited potential for the treatment of hyperlipidemia in current in vitro investigations. Generally, lead compounds with novel structures were developed in the present study for further design of lipid-lowering molecules by targeting PCSK9/LDLR PPI.

ProTide and cyclic phosphate ester are two successful prodrug technologies to overcome the limitations of nucleoside drugs, among which the cyclic phosphate ester strategy has not been widely used in the optimization of gemcitabine. Herein, we designed a series of novel ProTide and cyclic phosphate ester prodrugs of gemcitabine. Cyclic phosphate ester derivative 18c exhibits much higher anti-proliferative activity than positive control NUC-1031 with IC50s of 3.6-19.2 nM on multiple cancer cells. The metabolic pathway of 18c demonstrates that 18c's bioactive metabolites prolong its anti-tumor activity. More importantly, we separated the two P chiral diastereomers of gemcitabine cyclic phosphate ester prodrugs for the first time, revealing their similar cytotoxic potency and metabolic profile. 18c displays significant in vivo anti-tumor activity in both 22Rv1 and BxPC-3 xenograft tumor models. These results suggest that compound 18c is a promising anti-tumor candidate for treating human castration-resistant prostate and pancreatic cancer.

The limited regenerative potential of the optic nerve in adult mammals presents a major challenge for restoring vision after optic nerve trauma or disease. The mechanisms of this regenerative failure are not fully understood1,2. Here, through small-molecule and genetic screening for epigenetic modulators3, we identify DNA methyltransferase 3a (DNMT3a) as a potent inhibitor of axon regeneration in mouse and human retinal explants. Selective suppression of DNMT3a in retinal ganglion cells (RGCs) by gene targeting or delivery of shRNA leads to robust, full-length regeneration of RGC axons through the optic nerve and restoration of vision in adult mice after nerve crush injury. Genome-wide bisulfite and transcriptome profiling in combination with single nucleus RNA-sequencing of RGCs revealed selective DNA demethylation and reactivation of genetic programs supporting neuronal survival and axonal growth/regeneration by DNMT3a deficiency. This was accompanied by the suppression of gene networks associated with apoptosis and inflammation. Our results identify DNMT3a as the central orchestrator of an RGC-intrinsic mechanism that limits optic nerve regeneration. Suppressing DNMT3a expression in RGCs unlocks the epigenetic switch for optic nerve regeneration and presents a promising therapeutic avenue for effectively reversing vision loss resulted from optic nerve trauma or diseases.

The brain injury caused by cerebral ischemia-reperfusion is related to mitochondrial damage. Maintaining the normal function of mitochondria, promoting angiogenesis, protecting neuronal cells, and resisting oxidative stress are the keys to functional recovery after acute ischemic stroke. In this study, we established a middle cerebral artery occlusion (MCAO) model and investigated the effects of 1α,25-dihydroxyvitamin D3 (VitD or 1,25-D3) on mitochondrial function via the adenosine 5'-monophosphate-activated protein kinase (AMPK)/protein kinase B (AKT)/glycogen synthase kinase-3β (GSK-3β) signaling pathway in rats with cerebral ischemia-reperfusion injury. The neurological function and infarct size were measured in each group. Hematoxylin-eosin, neuronal nucleus, and Nissl staining procedures were conducted to observe the morphology and number of the cerebral cortical neurons. Western blotting was then used to analyze p-AMPK, vitamin D receptor (VDR), p-GSK-3β, p-AKT, P53, cytochrome C (CytC), TGF-β, and vascular endothelial growth factor (VEGF) in mitochondria. Immunofluorescence staining was used to observe the expression of CytC and caspase-3. Succinate dehydrogenase, ATPase, reactive oxygen species, and malondialdehyde were detected by kits. RT-qPCR was used to analyze TGF-β, VEGF, P53, and CytC mRNA. The results revealed that the cerebral infarct volume, neurological function score, apoptotic protein P53, CytC, caspase-3, reactive oxygen species, and malondialdehyde were significantly increased in MCAO rats. 1,25-D3 reduced the infarct size and neurological function score, activated VDR, upregulated TGF-β, p-AMPK, p-AKT, p-GSK-3β, VEGF, ATP, and succinate dehydrogenase, and downregulated P53, CytC, caspase-3, reactive oxygen species, and malondialdehyde. As an antagonist of VDRs, pyridoxal-5-phosphate could partially block the neuroprotective effect of 1,25-D3. In conclusion, 1,25-D3 activated AMPK/AKT/GSK-3β signaling and VDRs, inhibited P53, CytC, and caspase-3, increased TGF-β and VEGF, regulated mitochondrial metabolism, reduced neuronal apoptosis, promoted vascular growth, and exerted neuroprotective effects. These findings suggest that this signaling pathway may be an effective target for the treatment of ischemic stroke.

The goal of this work was to develop a sensitive and accurate method based on high performance liquid chro-matography with tandem mass spectrometry (LC-MS) detection for determining the concentration of the Sino-menine derivative SWX in plasma and tissue of rat. Chromatographic separation was achieved on an Agilent SB-C18 (2.1*50 mm, 2.7 mu m) column. The mobile phase consisted of acetonitrile (solvent A) and 0.2 % formic acid in water (solvent B) with gradient elution as follows: 0-3 min, 50-90 % A, 3 min-3.01 min, 90-50 % A, 90 % A in 3.01 min-10 min. The flow rate was set to 0.3 mL/min. The column temperature was 40 degrees C, and the sample injection volume was 10 mu L. The calibration curve had good linearity in the range of 10 ng/mL to 600 ng/mL. Biological samples were prepared by protein precipitation with acetonitrile. The area under the curve (AUC) and the peak drug concentration (Cmax) were linearly related to SWX dose. After oral administration of 25, 50 and 100 mg/kg SWX and intravenous administration of 0.5 mg/kg SWX, respectively, the absolute bioavailability of SWX was estimated as 12.4 %, 12.2 % and 10.6 %. SWX was widely distributed in heart, liver, spleen, lung, kidney, brain, breast, and fat, especially in lung, liver, and spleen. Distribution of SWX in brain suggested that it can pass the blood-brain barrier. The method established in this study has the advantages of high recovery and good reproducibility, and was suitable for the determination of the content of Sinomenine derivatives, providing a reliable scientific tool for carrying out pharmacokinetics research, providing a reliable scientific resource.

Aminopeptidase N(APN/CD13), a Zn2+-dependent ectopeptidase localized on the cell surface, is widely considered to influence the invasion of tumor cells. We found that boroleucine and dino-leucine borate exhibited a strong inhibitory effect on the enzyme activity of aminopeptidase N. The tested assay indicated that both compounds had an anti-proliferative effect on triple-negative breast cancer cells. Wound healing assay, migration test and matrigel-coated transwell assay showed that both boroleucine and dino-leucine borate inhibited the migration and invasion of breast cancer cells. Immunoblot analysis showed that both compounds down-regulated the expression of matrix metalloproteinase-2/9. In the capillary tube formation assay of human umbilical vein endothelial cells (HUVECs), dino-leucine borate showed better antiangiogenic activity than ubenimex even at a low concentration (10 μM). Moreover, compared with ubenimex, the anti-metastatic activity of dino-leucine borate in vivo was similar to or even better than that of ubenimex in the H22 pulmonary metastasis mouse model. In this paper, we found the novel APN inhibitors to markedly suppress the enzyme activity of APN and inhibit the migration and invasion of tumor cells in vitro and in vivo.

Tumor growth and metastasis are caused by many factors. The complexity means that a multi-target combination therapy strategy should be selected against tumor growth and metastasis. Here, cisplatin (CDDP) and bendazac (Ben) were designed as a novel NSAID-Pt(IV) nanoplatform, which is an effective weapon for combating tumor growth and metastasis.

Dysregulation or aberrant signaling transduction contributes to tumorigenesis. Targeting these abnormal signaling pathways becomes an effective anticancer strategy. However, feedback activation or crosstalk between signaling pathways drives adaptive drug resistance which causes failure of cancer therapy. In this review article, we summarized treatments that cause feedback activation of AKT, ERK, STAT3, EGFR, FGFR, and HER2/3 signaling pathways and the combination therapy to enhance anti-tumor effect or to overcome drug resistance, to explore the underlying mechanisms that define the protein molecules participated or regulated the feedback activation. In addition, we reviewed clinical trials that employ combination treatments to suppress feedback activation and improve therapeutic efficacy of cancer treatments.

本研究针对我校医学本科生药理学课堂教学中的实际问题，提出“五体同向”教学模式，即：以讲授法为本体，启发式教学为思体，生生、师生交互讨论为分体，情景演绎教学和案例教学法为源体，以网络平台软件为多媒体，五大方面相互融合，以学生为主导，变被动为主动，完成学生从复制记忆知识到实践应用创新的过渡，达到协同提升医学生学习质量和药理学课堂教学质量的目标。

Survival analysis models are necessary for clinical forecasting with data censorship. Implicitly, existing works focus on the individuals with higher risks while lower risk individuals are poorly characterized. Developing survival models to represent different risk individuals equally is a challenging task but of great importance for providing accurate risk assessments across levels of risk. Here, we characterize this problem and propose an adjusted log-likelihood formulation as the new objective for survival prognostication. Several models are then proposed based on the newly designed optimization objective function which produce risks that count individuals “equally” on risk ratios thus providing representative attention to individuals of varying risk. Extensive experiments on multiple real-world datasets demonstrate the benefits of the proposed approach.