Ferroptosis, a novel form of programmed cell death driven by iron-dependent lipid peroxidation, plays a crucial role in both disease treatment and microbial control due to its multi-level regulatory mechanisms. It mainly involves iron metabolism, lipid peroxidation, and antioxidant systems. In the field of disease treatment, ferroptosis is regarded as a highly promising therapeutic target because of its key role in autoimmune diseases, cancer, and cardiovascular diseases. This review systematically summarizes the core regulatory factors of ferroptosis, such as glutathione peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4) and their interaction networks, and deeply explores the application prospects of targeted intervention strategies based on ferroptosis signaling pathways in disease treatment and microbial control. Additionally, we also summarize the current issues faced by ferroptosis in practical applications and proposes strategies, such as nanodelivery, improved drug chemical stability and enhanced water solubility to optimize therapeutic efficacy. We aim to provide a theoretical basis and practical guidelines for exploring more targeted treatments using ferroptosis.

Attention-deficit/hyperactivity disorder (ADHD) is a common neurodevelopmental disorder in children and adolescents, and is clinically characterized by inattention, hyperactivity, and impaired impulse control. Despite extensive research, its etiology and pathogenesis remain incompletely understood. The dopamine (DA) deficiency theory constitutes a central framework in current ADHD studies. In-depth investigations of dopamine transporter (DAT) and dopamine receptor (DR) functions have led to the development of mainstream pharmacological treatments, which alleviate symptoms by enhancing DA concentrations and further support the essential role of the dopaminergic system in ADHD. Beyond DA transport and receptor signaling, recent findings suggest that impaired DA release may contribute to DA deficiency. The soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, composed of synaptosomal-associated protein 25 (SNAP-25), syntaxin-1A (STX1A), and vesicle-associated membrane protein 2 (VAMP2, also known as synaptobrevin 2), serves as the core molecular machinery mediating synaptic vesicle docking and membrane fusion at the presynaptic terminal. As the minimal apparatus driving DA vesicle exocytosis, the SNARE complex precisely regulates presynaptic DA release through coordinated vesicle fusion and recycling. Functional disruptions in SNARE complex assembly, disassembly, or its auxiliary regulatory proteins may hinder effective DA transmission, potentially contributing to DA deficiency and representing a novel molecular mechanism in ADHD pathogenesis. This review summarizes the current understanding of the SNARE complex and its regulatory network, emphasizing their potential roles in the pathogenesis and progression of ADHD, and offering theoretical insights into disease mechanisms and targeted therapeutic strategies.

With the continuous increase in global obesity prevalence, the impact of obesity on reproductive physiology has garnered widespread societal attention. As a metabolic disorder, obesity is typically accompanied by multiple abnormal physiological phenomena, such as excessive adipose accumulation and exacerbated inflammatory responses, which severely compromise the reproductive health of humans and animals. Reproductive damage induced by obesity involves a series of complex biochemical reactions and in vivo metabolic pathways, manifesting as impaired male sperm quality and female fertility. To better understand the relationship between obesity and reproductive physiology, this review summarizes the reproductive injuries caused by obesity and their underlying mechanisms. In the obese state, conditions such as oxidative stress, insulin resistance, and hyperinsulinemia are induced, with adipokines (leptin, adiponectin, resistin, etc.) and inflammatory factors (TNF-α, IL-6, IL-1β, etc.) interacting synergistically to affect the reproductive system. Oxidative stress activates the MAPK and NF-κB pathways, interfering with insulin signaling, while chronic inflammation leads to adipocyte secretory disorders and disrupts the hypothalamic-pituitary-gonadal regulatory axis. Studies have shown that obese males exhibit significantly decreased testosterone levels and impaired sperm quality, whereas obese females suffer from reproductive hormone imbalance, ovulation disorders, and polycystic ovary syndrome. This review discusses how obesity-induced metabolic disorders lead to impaired reproductive physiology in both males and females, along with the underlying mechanisms, providing a theoretical basis for the prevention and treatment of obesity-related reproductive disorders in the future.

The structure and function of mitochondria and endoplasmic reticulum (ER) are important for maintaining cellular homeostasis. It has been found that the interaction between mitochondria and ER is involved in the occurrence and development of a variety of diseases. The mitochondria-associated ER membrane (MAM) is a membrane contact site between the ER and mitochondria, and is an important communication center between organelles in eukaryotic cell. Calcium channels on the ER side and the mitochondrial side are crucial in the calcium transport process in MAM. The interaction between ER and mitochondria controls mitochondrial biological function and cell survival through calcium transport regulation, and are involved in the occurrence and development of various pathologic process. On the one hand, MAM regulates calcium transport, which is involved in the modulation of various cellular survival and death processes. It plays a profound regulatory role in the damage of tumor cells, neuronal cells, cardiomyocytes, endothelial cells and nucleus pulposus cells through different key molecules within MAM.On the other hand, the regulation of MAM in calcium transport is crucial in the development of mitochondrial dysfunction in Hepa 1-6 cells, the synthesis and secretion of pancreatic β-cells and amyotrophic lateral sclerosis. In addition, MAM also affects cellular transcription processes by regulating calcium transport, thereby exerting significant regulatory effects on angiogenesis and breast cancer. This paper reviews the structural features and pathophysiologic role of calcium transport regulation of MAM, and expects to provide new horizons for prevention and treatment of related diseases targeting MAM.

Occult hepatitis B virus (HBV) infection (OBI) represents a potential reservoir for HBV transmission, capable of spreading through routes such as blood transfusion. Additionally, OBI can contribute to the chronic progression of hepatitis B-related diseases, sustaining a state of chronic HBV infection. In individuals with compromised immune function or undergoing immunosuppressive therapy, OBI may lead to HBV re-activation, potentially triggering severe liver conditions such as acute hepatitis or liver failure. As a result, OBI poses a significant public health challenge, profoundly impacting the health and well-being of affected populations and complicating HBV infection control efforts in China. Clinically diagnosing OBI remains challenging, but its hallmark is serum hepatitis B surface antigen negative and the presence of HBV covalently closed circular DNA (cccDNA) in the liver. With the increasing focus on achieving functional cure for chronic hepatitis B, both domestic and international guidelines have refined functional cure. Notably, these guidelines acknowledge that cccDNA may persist in the liver tissue of individuals who have achieved functional cure, suggesting resemblance of an occult infection state. Here, we provide a comprehensive overview of OBI, including its definition, classification, public health implications, underlying mechanisms, and clinical reactivation. By updating the understanding of OBI, we aim to raise awareness among clinicians and public health professionals regarding the significance of OBI in the current context and encourage greater attention to this population.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by impairments in social interaction, communication, and repetitive behaviors. Accumulating evidence suggests that neuroimmune inflammatory responses may contribute to the pathogenesis of ASD. Within the central nervous system, microglia, as the key innate immune cells, play a pivotal role in shaping the neuroimmune inflammatory microenvironment. This review systematically synthesizes findings regarding alterations in microglial number and morphology across two major categories of ASD mouse models, considering both genetic and environmental dimensions. Specifically, it examines changes in dendritic spine density and neurotransmission function in gene mutation-induced models and environmentally triggered models, such as maternal immune activation models. Furthermore, this article highlights comparative analyses of shared mechanisms, including the interleukin-17 receptor A signaling pathway and the mammalian target of rapamycin signaling pathway, between MIA models and genetically induced BTBR T⁺Itpr3^tf/J mouse models. Building on these insights, the review elaborates on cytokine dysregulation in abnormally activated microglia, mitochondrial oxidative phosphorylation dysfunction, and elevated reactive oxygen species levels within ASD mouse brains, elucidating their implications for cellular function. Finally, the article summarizes how microglia influence neurodevelopment through neurogenesis and synaptic formation and function, exploring potential pathways underlying autism-like behavioral phenotypes and identifying novel therapeutic targets for clinical ASD intervention.

Monkeypox is a viral zoonotic disease, and there is currently a lack of safe and effective vaccines against the monkeypox virus. Therefore, screening and developing vaccine candidates is of significant practical importance. With the rapid advancement of molecular biology and plant genetic engineering, plant bioreactors offer promising potential for producing vaccine proteins due to their advantages, including safety, cost-effectiveness, and scalability. In this study, we focused on the monkeypox protein B6R. The recombinant expression plasmid pFolia40108-B6R-Fer was successfully constructed using amplification, enzyme digestion, and flexible linker tandem ferritin technology. A complete transient expression system in Nicotiana benthamiana and a purification system for the recombinant monkeypox protein were established. The optimal expression time was determined to be 12-14 days, with a final purified protein concentration of approximately 1 mg/mL and a yield of 0.85 mg/kg fresh weight. The purified B6R-Fer recombinant protein self-assembled into spherical virus-like particles (VLPs) with an average particle size of 24 nm. The B6R-Fer recombinant protein from this study shows promising potential for use in the development and screening of plant-derived monkeypox vaccine candidates.

Every year, up to 800 million tons of hydrocarbons enter the environment globally, most of which are alkanes. Due to the inactive property and the high freezing points, alkanes have caused serious problems on environmental ecology and oil recovery. Alkane monooxygenase (AlkB) is a transmembrane metalloproteinase, and belongs to the membrane-bound fatty acid desaturase (FADS) family, which is able to convert straight-chain alkanes into the corresponding primary alcohols during the first step of alkane degradation mediated by microorganisms. Thus, AlkB plays a crucial role in the global carbon cycle and bioremediation of oil pollution. In this paper, the characteristics, structure, active site, catalytic mechanism, and the construction of recombinant bacteria of AlkB from different microorganisms were reviewed. In addition, the important significance of AlkB for environmental remediation and oil extraction was also emphasized, which would provide new clues for the bioremediation of hydrocarbon-contaminated sites and improvement of oil recovery rate by AlkB.

The MYB protein family is one of the largest transcription factor families in plants, widely involved in growth and development, stress responses, and secondary metabolism. However, the MYB protein family members in Lonicera japonica have not been identified yet. In this study, a genome-wide identification and analysis of the MYB protein family in L. japonica was conducted using bioinformatics methods, covering basic physicochemical properties, phylogenetic trees, gene structures, conserved motifs, and cis-acting elements. Additionally, the subcellular localization of MYB6, MYB106d, and MYB114 proteins was detected through the construction of pCAMBIA1300-GFP fusion vectors and transient transformation in Nicotiana benthamiana leaves. The results showed that a total of 147 LjMYB genes were identified, belonging to 17 subfamilies (S1-S17). The encoded amino acid lengths ranged from 52 to 1 060 AA, isoelectric points from 4.42 to 11.52 pI, and the number of exons from 1 to 13, with molecular weights ranging from 6 086.3 to 119 075.08 kD. MEME analysis revealed that the number and distribution of motifs in different MYB proteins varied, while the structural features within the same subfamily were similar. The analysis of cis-acting elements indicated that the promoter regions contained light-responsive, hormone-responsive, and biotic and abiotic stress-related elements and binding sites. Chromosome distribution showed significant genome doubling of MYB genes (possibly related to chromosome evolution doubling). The collinearity analysis of MYB genes between L. japonica and Arabidopsis thaliana revealed 121 pairs of homologous genes distributed across all A. thaliana chromosomes, demonstrating evolutionary conservation. Expression profile analysis indicated that L. japonica MYB genes played different roles in growth and development and had varying sensitivities to different light intensities. Subcellular localization showed that LjMYB6, LjMYB106d, and LjMYB114 were all localized in the nucleus. In conclusion, the MYB protein family in L. japonica has diverse biological characteristics and may be involved in growth and development, hormone regulation, and biotic and abiotic stress responses.

In order to investigate the teaching effectiveness of artificial intelligence (AI) in molecular biology, this study selected students from the Animal Medicine major of the College of Animal Science and Technology at Hebei North University in 2022 and 2023 as research subjects. There was no significant difference in professional foundation, admission scores, and other aspects between the two grades, and they were taught by the same lecturer to ensure the reliability of research results. The 2022 students will adopt the traditional teaching mode, while the 2023 students will implement the AI-enabled teaching mode, which includes four stages: pre class exploration, in class assistance, post class learning support, and teaching reflection and improvement. Before class, the teaching team pushes teaching videos of various knowledge points in the course and relevant preview materials organized by the AI system to students to complete self-learning, and use AI systems to track students' learning difficulties. In class, the teacher uses various teaching methods such as case teaching, group discussions, AI animation demonstrations and virtual experiments, etc.. Thus, they provide in-depth explanations of key contents based on the feedback data from the intelligent learning companion AI system, promoting students' understanding of the knowledge. After class, the AI system generates personalized learning plans for students and provides different levels of learning resources to broaden their horizons. At the same time, the AI system provides teachers with students' learning data analysis reports, and teachers can adjust and optimize their teaching plans accordingly. Research has found that students in the 2023 AI-empowered teaching class have significantly higher satisfaction in multiple dimensions such as learning interest, understanding and mastery of knowledge points, and cultivation of scientific research thinking than those in the 2022 traditional teaching class. In terms of student participation in comprehensive activities, the proportion of 2023 students participating in subject competitions and innovation and entrepreneurship activities has significantly increased. In terms of academic performance, the mid-term, laboratory, and final grades of 2023 students are higher than those of 2022 students, with a significant increase in the excellence rate and a significant decrease in the failure rate. The results indicate that the application of AI technology in molecular biology teaching has stimulated students' interest in learning, helped them better understand and master knowledge, significantly improved their academic performance. In sum, it has a positive impact on improving teaching quality.

Multiple myeloma (MM) is a hematologic malignancy characterized by clonal proliferation of plasma cells within the bone marrow, with pathological features including abnormal secretion of monoclonal immunoglobulins, osteolytic bone disease, and multi-organ dysfunction. Despite significant advancements in therapeutic approaches that have markedly extended patient survival, primary drug resistance and relapse remain major obstacles to clinical cure. The pathogenesis and progression of MM are intricately regulated by the bone marrow microenvironment (BMME), a dynamic network composed of diverse cellular and non-cellular components. The BMME not only supports the survival and proliferation of MM cells but also plays a pivotal role in disease progression by modulating bone metabolic homeostasis, mediating immune escape, and promoting drug resistance. In recent years, groundbreaking therapeutic strategies targeting the BMME have emerged, including immunomodulatory drugs, bispecific antibodies, CAR T-cell therapies, and microenvironment-modulating agents. These approaches have significantly improved objective response rates and survival outcomes in relapsed/refractory MM by disrupting cytokine signaling, reprogramming the immunosuppressive microenvironment, or inhibiting tumor-stromal interactions. However, challenges such as drug resistance, treatment-related toxicity, and tumor heterogeneity persist in clinical practice. This review systematically delineates the roles of BMME components in MM pathogenesis, analyzes the molecular mechanisms underlying MM cell-BMME interactions, and explores innovative strategies to enhance therapeutic efficacy and prognosis through targeted modulation of the BMME. These insights provide a foundation for developing novel therapeutic paradigms aimed at overcoming current limitations in MM treatment.

UBE2O is a distinctive ubiquitin-conjugating enzyme characterized by its large size (1 292 residues) and dual E2/E3 enzymatic activities, enabling diverse ubiquitylation types. Unlike typical E2 enzymes (150~200 residues), UBE2O’s multifunctionality allows it to regulate substrate degradation, subcellular localization, and functional modulation. Emerging studies highlight its critical roles in protein quality control, erythroid differentiation, metabolic regulation, and maintenance of circadian rhythm. Dysregulation of UBE2O is implicated in various diseases, including cancers, neurodegenerative disorders, and metabolic diseases. This review extensively discusses the unique structural features, diverse biological functions, and pathological roles of UBE2O, as well as its therapeutic potential for associated diseases.

Multiple myeloma (MM) is a hematologic malignancy characterized by the malignant proliferation of plasma cells. Currently, proteasome inhibitors (PIs),immunomodulatory drugs (IMiDs), and autologous hematopoietic stem cell transplantation (ASCT) are the primary approaches used to improve the survival outcomes of MM patients. However, as treatment advances, most patients still face refractory and relapsed diseases, and the path to a cure remains challenging. In recent years, monoclonal antibodies (mAbs), chimeric antigen receptor T cells (CAR-T),antibody-drug conjugates (ADCs), and bispecific antibodies (BsAbs) have all demonstrated promising efficacy in clinical studies and applications. These antibody and cell-based immunotherapies have brought new hope to patients with refractory and relapsed MM (RRMM). Each type of immunotherapy offers unique advantages in activating immune cells, targeting tumors, and improving patient outcomes, yet they also encounter challenges related to safety and resistance. In the future, with continuous advancements in molecular biology and antibody engineering, novel immunotherapeutic products are expected to achieve synergistic effects through combination strategies, thereby providing longer survival benefits and improved life quality for RRMM patients. This article aims to review the latest progress in immunotherapy for MM, systematically discussing the mechanisms of action, current clinical applications, and future development trends of mAbs, CAR-T, ADCs, and BsAbs. It is intended to serve as a comprehensive reference for researchers and clinicians and promote advancing precision immunotherapy for MM.

Multiple myeloma (MM) is the second most common hematological malignancy, predominantly affecting the elderly. With the development of an aging society in China, the incidence of MM has been steadily increasing, becoming a significant concern for public health and a considerable socioeconomic burden. Over the past two decades, breakthroughs have been achieved in both clinical management and basic research on MM. However, the field now stands at a crossroads for the next phase, facing numerous developmental directions and challenges.

Biotoxins are widely distributed in animals, plants, and microorganisms, functioning as “chemical weapons” of proteins, peptides, or small molecules that evolved for predation, defense, and competition. Compared with general chemical toxins, biotoxins exhibit high potency, strong specificity, and remarkable molecular diversity. While posing potential threats to human health, they also provide unique value in elucidating disease mechanisms and inspiring drug development. Representative drugs such as captopril, botulinum toxin, ziconotide, and GLP-1 receptor agonists mark milestones in the therapeutic application of toxins. To date, over one hundred biotoxin-derived drug candidates have entered clinical trials across multiple major diseases. However, this field still faces challenges, including low efficiency in resource discovery, limited structural and mechanistic insights, inherent toxicity, and constraints in synthesis and modification technologies. Looking forward, advances in multi-omics, artificial intelligence, and synthetic biology will drive efficient toxin discovery, detoxification strategies, and precision applications, ultimately promoting a closed-loop progression from basic research to clinical translation.

tRNA is one of the RNA molecules with the most diverse post-transcriptional modifications. It not only functions as an adaptor molecule transporting amino acids during translation but also relies on its extensive post-transcriptional modifications to regulate gene expression, thereby influencing numerous biological processes. These modifications are dynamically regulated by tRNA methyltransferases and demethylases, which collaboratively maintain a balance—the former catalyze methyl group addition, while the latter remove methyl groups, ensuring reversible control. Mutations or dysregulation of these enzymes are closely associated with tumorigenesis and other diseases, constituting a major research focus. They promote tumor progression through two distinct pathways: cytoplasmic tRNA (ctRNA) methylation enhances the translation of oncogenes, whereas mitochondrial tRNA (mtRNA) methylation optimizes mitochondrial protein synthesis to reprogram energy metabolism. From the dual dimensions of"enzyme supply" and"energy provision", tRNA methylation establishes a material foundation for tumor cell growth and directly contributes to cell cycle dysregulation. The cell cycle, an orderly process governing cell division, is tightly controlled by checkpoint proteins to guarantee accurate genetic information transmission. Notably, aberrant cell cycle activation is not only a hallmark of cancer but also a pivotal therapeutic target. tRNA methyltransferases and demethylases exert multidimensional control over both cell cycle progression and metabolic adaptation. Elucidating the mechanisms underlying tRNA modification mediated cell cycle regulation will not only accelerate the discovery of novel therapeutic targets but may also overcome the constraints of conventional cell cycle targeted therapies. Within this context, we systematically review the molecular mechanisms by which tRNA methylation-modifying enzymes regulate the cell cycle, with an emphasis on their translational potential.

Orphan G protein-coupled receptor 35 (GPR35) is a GPCR that modulates lipid metabolism and exerts cell-type-specific metabolic control in distinct adipocyte populations. In white adipocytes, GPR35 regulates both lipolysis and lipogenesis, and is tightly linked to insulin sensitivity and inflammatory responses. In brown adipocytes, it orchestrates lineage commitment and energy expenditure by activating uncoupling protein 1 (UCP1) and regulator of G-protein signaling 14 (RGS14). In beige adipocytes, GPR35 promotes “browning”, fine-tunes mitochondrial function, and governs thermogenic capacity. Within adipose tissue, GPR35 further modulates immune, neuronal, and vascular compartments—alleviating inflammation and tuning blood flow—to orchestrate intercellular crosstalk that ultimately shapes adipocyte metabolism. Clinical studies of obesity, type 2 diabetes, and MASLD have implicated GPR35 in lipolysis, energy expenditure, insulin sensitivity, and inflammatory tone. Nevertheless, its precise mechanisms remain incompletely understood, and the selectivity of candidate ligands requires optimization, leaving therapeutic translation still fraught with challenges. Future work should leverage molecular docking, deep-learning models, and genome-editing technologies to clarify GPR35 function, validate drug specificity, and develop innovative therapeutic strategies for metabolic diseases.

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling pathway, as a crucial component of innate immunity, plays a pivotal role in anti-tumor immunity. cGAS recognizes aberrant endogenous or exogenous DNA and catalyzes the synthesis of the second messenger cGAMP, which activates the STING protein. This triggers the TBK1-IRF3/NF-κB signaling axis to induce the secretion of cytokines such as interferon-I and other cytokines, which activates dendritic cells, enhances natural killer cell function, promotes T cell responses, and strengthens immune surveillance, thereby inhibiting tumor progression. However, tumor cells can perform immune surveillance via multiple mechanisms, including downregulation of cGAS/STING expression, accelerated STING protein degradation, or inhibition of cGAMP synthesis and signal transduction, leading to the formation of an immunosuppressive microenvironment and facilitating tumor immune escape. Current strategies to target cGAS-STING pathway activation primarily involve the development of STING agonists, optimization of delivery systems, and combination therapies with PD-1/PD-L1 inhibitors, radiotherapy, or chemotherapy to synergistically enhance antitumor immune responses. Despite promising prospects, this field still faces challenges such as systemic toxicity of STING agonists, tumor microenvironment heterogeneity, and drug resistance mechanisms. This review summarizes the function and molecular mechanisms of the cGAS-STING pathway, its role in tumor immunity, the development in STING agonists and delivery systems, as well as its potential for combination with immune checkpoint inhibitors, radiotherapy, and chemotherapy. It also discusses the challenges and optimization directions for targeting the cGAS-STING pathway, providing theoretical foundations and insights for developing more effective tumor immunotherapy strategies.

O-glycosylation (including mucin-type O-glycosylation and O-GlcNAcylation), as a critical post-translational modification (PTM), regulates protein function, stability, and subcellular localization through the addition of glycan chains to serine or threonine residues, which participates in cellular signaling, metabolic regulation, and stress responses. DNA damage refers to the disruption of genomic integrity caused by endogenous factors (e.g., metabolic byproducts, replication errors) or exogenous agents (e.g., radiation, chemical substances), leading to carcinogenesis, aging, and genetic disorders. To counteract DNA lesions, organisms have evolved the DNA damage response (DDR) system, which orchestrates complex protein networks to detect DNA damage and facilitate repair processes. Emerging evidence indicates that O-glycosylation can modulate DDR by influencing the activity, localization, and interactions of DNA repair-associated proteins. However, the precise mechanisms underlying O-glycosylation-mediated DDR remain to be clarified. This review systematically summarizes: (1) the biosynthetic pathways of mucin-type O-glycosylation and O-GlcNAcylation, the cascade reactions in DDR; and (2) current research advances regarding O-glycosylation in tumor-associated DDR. Furthermore, we propose novel mechanistic perspectives and therapeutic strategies targeting O-glycosylation-mediated DDR dysregulation in malignancies, aiming to provide a theoretical basis for tumor treatment.

This study aims to explore the efficacy and mechanisms of achyranthes bidentata extract on chondrocytes pyroptosis and cartilage injury in knee asteoarthritis in a rat model. A rat KOA model was constructed using “medial collateral ligament transection (MCLT) + partial meniscectomy (PM)”method. Hematoxylin and eosin (H&E) staining and safranin O/fast green staining were performed to estimate the pathological status of the damage. TUNEL staining was performed to detect the chondrocytes pyroptosis. Micro-CT was used to assess bone damage and KOA severity. LPS-treated chondrocytes extracted from rat knee cartilage were used as an in vitro model. Cytotoxicity and cell viability were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) kit. The levels of inflammatory cytokine and stromal proteins were quantified by ELISA and Western blot assays. Caspase-1 activity was evaluated by flow cytometry. The level of NLRP3 in chondrocytes was examined by immunofluorescence. Our results revealed that ABE inhibits rat chondrocyte injury and pyroptosis by reducing the levels of LDH (P<0.01), IL-18 (P<0.01), IL-1β (P<0.01), MMP-1 (P<0.01), and MMP-13 (P<0.001) at the same time increasing collagen II (P<0.001) expression. Furthermore, ABE suppressed the NLRP3 (P<0.001) inflammasome and Caspase-1 (P<0.001) signaling pathways to alleviate pyroptosis. The inhibitory effects of ABE on chondrocyte pyroptosis were mediated by P2X7R regulation. P2X7R overexpression suppressed the above positive changes caused by ABE. We further confirmed that ABE could prevent the pathological conditions in the rat KOA model by suppressing P2X7R/NLRP3-mediated pyroptosis. In conclusions, ABE suppressed KOA progress by repressing P2X7R/NLRP3 signaling-mediated pyroptosis in chondrocytes and rat KOA models, indicating that ABE may act as a promising medicine for KOA treatment.

Multiple myeloma (MM) is a hematologic malignancy originating from plasma cells in the bone marrow. In recent years, the use of novel drugs and hematopoietic stem cell transplantation (HSCT) has significantly improved the prognosis of MM patients. However, MM remains challenging to cure and is prone to relapse and resistance. For patients with relapsed/refractory multiple myeloma (RRMM), there is an urgent need to explore novel therapeutic approaches. Bispecific antibodies (BsAbs) are a promising class of immunotherapeutics that functions by simultaneously binding to tumor cell antigens and endogenous effector cells, thereby forming an immunological synapse. This interaction facilitates the activation of effector cells, leading to the targeted lysis of tumor cells. Numerous clinical studies have demonstrated the significant efficacy of BsAbs, either as monotherapy or in combination with other treatment regimens, in patients with RRMM. This article summarizes the mechanisms of action, efficacy, and safety of BsAbs, and discusses optimal sequencing strategies in immunotherapy, aiming to provide new perspectives for the treatment of MM.

Most recently, Quake proposed the “cellular dogma” in which the central dogma, cell theory and theory of evolution should be integrated to understand a major conceptual challenge for biology in the next decade. However, the “cellular dogma” remains to be developed theoretically and experimentally. Quantum biology is an interdisciplinary field to study quantum effects in dynamic structures of biological molecules and energy transfer, and further understand the quantum mechanics of chemical processes and the origin of life by using quantum theories and methods. We wonder whether quantum biology could explain the basic nature of life in the most general sense. Herein, we first provide a brief review on the development of quantum mechanics and the conceptions in quantum biology, then we summarize the present theories of quantum biology and discuss a possible prospectives. We cover the following contents of quantum biology: Quantum tunneling due to a decrease of proton stability by hydrogen bond breakage in DNA replication and repair cause permanent mutations with evolutionary impacts; the tunneling due to conformational short-range motion and static environment optimization in enzyme molecules is found to promote enzymatic reactions by reduction of the effective barrier height; the activated electrons are in a phase-synchronized superposition state between multiple pigment molecules, enhancing the conversion of light energy into chemical energy with high efficiency by optimizing the energy transfer path through phase-length interference; quantum compasses in bird migration lead to sense the way by detecting weak magnetic fields through electron transfer and successive conformational changes in cryptochrome; collapses of conformational state of microtubules in the nervous system due to quantum coherence, superposition, entanglement and Orchestrated Objective Reduction (Orch-OR) collapse can sense and transmit neuro-information. Finally, the paper also discusses the possibilities and prospectives in experimental evidence for quantum mechanical simulation of life activities, whether the “observer effect” may provide new ideas for explaining the origin of life, and whether the Copenhagen interpretation could explain the basic nature of life.

The aim of this study is to explore a new path of empowering ideological and political education in courses with digital intelligence technology, therefore to improve the teaching quality of biology courses in universities, promote the comprehensive implementation of the fundamental task of cultivating morality and talents, and continuously improve the level of "three pronged education". By using the biology course "Immunology" as a practical carrier, we have constructed a dual-driven framework of "ideological and political guidance+digital intelligence empowerment" utilizing digital tools to further highlight pedagogical feature enhancement, curricular optimization, innovative teaching models, multidimensional assessment, and enhanced teaching refinement. The empowerment of digital intelligence in the ideological and political construction of the course "Immunology" has achieved notable progress. Students’ professional abilities and ideological and political literacy have significantly improved, and the proportion of students with excellent grades has been increasing year by year (81.93% of students with good grades or above in 2024), demonstrating outstanding performance in various social practice activities; The course exhibits excellent demonstration and radiation effects on campus, and the teaching results have been supported by the Ministry of Education and provincial-level education reform projects. This study serves as a practical example for the digital transformation of ideological and political education in higher education courses, providing reference and innovative insights for implementing ideological and political elements in other courses of biology majors.

Breast cancer (BRCA) remains one of the leading causes of cancer-related deaths worldwide due to its high rates of metastasis and recurrence, making it crucial to explore its underlying molecular mechanisms. Our previous study demonstrated that miR-326 inhibits BRCA progression by targetingEPH receptor B3(EPHB3). This study further explores the molecular mechanism by which long non-coding RNAs (LncRNAs) regulates BRCA progression via the competing endogenous RNA (ceRNA) mechanism, in which it competes with EPHB3 for miR-326 binding. Bioinformatics analysis identified LncRNA Small Nucleolar RNA Host Gene 12 (SNHG12) as a potential miR-326-binding molecule. SNHG12 was found to be significantly upregulated in BRCA tissues, exhibiting a negative correlation trend with miR-326 and a positive correlation trend with EPHB3, suggesting its potential involvement in the ceRNA regulatory network. Nuclear-cytoplasmic fractionation assays revealed cytoplasmic localization of SNHG12, while dual-luciferase reporter assays confirmed its direct binding to miR-326. Functional experiments demonstrated that SNHG12 knockdown significantly suppressed BRCA cell proliferation, invasion, and migration, while miR-326 inhibition reversed these effects. Furthermore, miRNA pulldown assay revealed significant enrichment of SNHG12 and EPHB3 in the miR-326 pulldown products, indicating direct binding between them. Western blotting and rescue experiments revealed that SNHG12 upregulates EPHB3 expression by sponging miR-326, thereby promoting the malignant behaviors of BRCA cells. Collectively, this study revealed that LncRNA SNHG12 promotes BRCA progression by regulating the miR-326/EPHB3 axis through a ceRNA mechanism. The SNHG12/miR-326/EPHB3 pathway may represent a promising target for the molecular diagnosis and targeted therapy of BRCA.

Non-alcoholic fatty liver disease (NAFLD) is an increasingly serious chronic liver disease worldwide, with complex pathogenesis and many challenges in diagnosis and treatment. In recent years, genome-wide studies have revealed the important roles of epigenetic modifications in the development of NAFLD, especially the involvement of imprinted genes. The parental origin effect of NAFLD suggests that imprinted genes play a key role in its pathogenesis. The Dlk1-Dio3 gene cluster, as one of the largest clusters of imprinted genes, has become a focus of research because of its central role in embryonic development and metabolic regulation. This review explores the structure and function of the Dlk1-Dio3 gene cluster and its potential role in NAFLD pathogenesis. This gene cluster plays a key role in the “second strike” of NAFLD through a complex regulatory network that affects biological processes such as lipid metabolism, glucose metabolism, inflammatory response and oxidative stress in the liver. Specifically, DLK1 acts as a negative regulator, inhibiting adipocyte differentiation and thus reducing hepatic lipid accumulation, while DIO3 promotes adipocyte differentiation and increases hepatic lipid accumulation by regulating thyroid hormone conversion. In addition, the Dlk1-Dio3 gene cluster regulates lipid metabolism by modulating multiple microRNAs (e.g. miR-370, miR-122, etc.). miR-370 exacerbates lipid accumulation by inhibiting CPT1α; miR-122 up-regulates SREBP-1c and promotes fatty acid synthesis; and miR-379/410 clusters increase lipid scavenging capacity by decreasing lipid accumulation. Long non-coding RNA MEG3 also plays an important role in NAFLD. meg3 promotes fatty acid oxidation and reduces lipid droplet accumulation by up-regulating SIRT6, and attenuates lipid synthesis by inhibiting the Wnt/mTOR signaling pathway through binding to miR-21. In terms of insulin resistance, DLK1 inhibits gluconeogenesis and promotes fatty acid oxidation by activating the PI3K/Akt/mTOR pathway, thereby reducing hepatic lipid burden. DIO3, on the other hand, affects insulin sensitivity by regulating thyroid hormones and promotes the development of NAFLD. Meanwhile, the Dlk1-Dio3 gene cluster also plays an important role in regulating oxidative stress and inflammatory responses, and DLK1 attenuates hepatic oxidative stress injury by inhibiting inflammatory factor expression and activating antioxidant signaling. Taken together, the Dlk1-Dio3 gene cluster plays a multidimensional role in the occurrence and development of NAFLD, providing potential biomarkers and therapeutic targets.

Rhabdomyosarcoma (RMS) is one of the most common malignant soft-tissue tumors in children and adolescents, characterized by a blockade of skeletal-muscle differentiation. Here we summarize basic studies and key advances in the field. We cover three aspects in RMS: differentiation control, cell-cycle regulation, and signaling networks. During early differentiation, Myogenic Differentiation 1 (MyoD) heterodimerizes with E-proteins and initiates muscle-lineage gene programs together with muscle-specific miR-206 and selected lncRNAs. During late differentiation, Myogenin orchestrates myoblast fusion and myotube maturation, while its activity and stability are modulated by upstream regulators including the miR-1-TRPS1 axis, Arp5, and IL-4/STAT6. At the cell-cycle level, the p21/p27 and Rb-E2F axes promote G₁ arrest to license differentiation. When these signaling pathways are disrupted, tumor cells maintain high proliferative activity, but cell differentiation is blocked. At the signaling level, aberrant activation of PI3K/AKT/mTOR and MAPK/ERK, together with differentiation-suppressive pathways such as Notch and Hedgehog, interconnect to form a self-sustaining “proliferation-de-differentiation” loop. Mechanistically informed strategies—including inhibition of aberrant pathways, correction of epigenetic and non-coding RNA imbalances, restoration of MyoD/Myogenin function, and CRISPR-based precision interventions—show potential to induce differentiation and restrain tumor progression. Remaining challenges include unclear causal relationships among pathways and subtype-specific drivers, a paucity of predictive biomarkers, and insufficient definition of therapeutic windows and resistance dynamics for combination regimens. Future work should leverage single-cell and spatial omics to integrate epigenetic and post-transcriptional layers, reconstruct actionable differentiation networks, and—under biomarker guidance—develop and stratify cross-pathway combination strategies to advance RMS differentiation therapy toward precision and clinical translation.

The engineering application of microbially induced carbonate precipitation (MICP) is limited by pH-dependent conformational dynamics of urease. Focusing on the α-subunit urease from Sporosarcina pasteurii, this study integrated conductivity experiments and constant-pH molecular dynamics simulations to analyze active site conformational dynamics and catalytic function across pH 3-11. Results showed that under neutral conditions (pH 7-8), key histidine residues (HIS139/HIS249) exhibited minimal displacement (<0.5 Å), the longest hydrogen bond lifetime (>8 ps), highest conformational stability (root mean square deviation, RMSD: 0.15-0.18 nm), and optimal catalytic activity (conductivity change rate: 0.03 mS/cm·min^-1, CaCO₃ precipitation: 3.84 g). Extreme pH (pH 3/11) induced structural collapse (displacement up to 1.8 Å) and complete activity loss. Simulations revealed that neutral pH stabilizes a protonation-dependent cooperative allosteric network by maintaining active site cavity volume (~120 ų) and moderate conformational coherence (correlation coefficient ~0.8). This work deciphers the molecular mechanism of pH-regulated urease dynamics through protonation states, providing theoretical support for MICP applications in acidic mine tailing remediation and alkaline soil stabilization.

Wound healing is a dynamic physiological process involving haemostasis, inflammation, proliferation and tissue remodeling. Skin injuries, such as diabetic foot ulcers, venous ulcers, and pressure ulcers, are difficult to heal and impose a serious physical and psychological burden on patients, and traditional treatments are difficult to address such problems. In recent years, Chinese herbal medicine-derived extracellular vesicles (CHMEVs) have shown promising potential in the field of wound repair and drug delivery due to their excellent biocompatibility, low immunogenicity and high safety. CHMEVs are nano-like particles enriched with herbal small molecule compounds, proteins and metabolites, and are able to cross biological barriers and effectively regulate intercellular communication to promote tissue repair. In this review, the isolation and extraction methods of CHMEVs and their roles in wound repair are reviewed, and the prospects and challenges of their clinical applications are discussed. The combination of engineered modification and biomaterials of CHMEVs further expands their potential for application in precision medicine and provides a new idea for the treatment of hard-to-heal wounds in the future.

Lung cancer poses a serious threat to global public health security. Chemotherapy, as the main strategy for cancer treatment, faces challenges such as high toxicity and drug resistance. Anticancer peptides have the potential of being developed into new anticancer drugs due to their advantages of broad-spectrum anticancer activity, rapid action, and difficulty in generating drug resistance, but they also face shortcomings such as weak activity and strong toxic side effects. The weakly acidic microenvironment of tumors (pH 6.5-6.8) provides a good idea for the design of anticancer peptides of high-efficiency and low-toxicity. Previously, we designed the acid-sensitive antibacterial peptide pHly-1 using the wolf spider (Lycosa singoriensis) toxin Lycosin-I as a template. In this study, we found that pHly-1 also had acid-sensitive anticancer activity. Further alanine scanning analysis of pHly-1 was carried out, and we obtained a mutant pHTP-2 with better acid sensitivity, whose IC₅₀ (half maximal inhibitory concentration) against A549 cells was 15.68 μmol/L at pH 6.6 and was greater than 100 μmol/L at pH 7.4. At pH 6.6, pHTP-2 could act on various lung cancer cell lines and induce the death of A549 cells by rapid lysis; at pH 7.4, 500 μmol/L pHTP-2 had weak toxicity to red blood cells (the hemolysis rate was approximately 38%) and primary myocardial cells (the inhibition rate was 49.7%, with P< 0.05). Analysis of its charge, particle size, morphology, and secondary structure showed that at pH 6.6, the histidine in the sequence of pHTP-2 was protonated, increasing the positive charge (P<0.01), decreasing the hydrated particle size (P<0.05) and forming an α-helical structure to induce membrane lysis of A549 cells. At pH 7.4, it was deprotonated, the positive charge decreases, a β-sheet structure was formed and self-aggregation occurred, limiting its effect on the A549 cell membrane and showing weak activity. In summary, pHTP-2 could respond to the weakly acidic microenvironment of tumors to exert selective cytotoxic activity, effectively overcoming the shortcomings of anticancer peptides such as low efficiency and high toxicity. Our findings suggest that it is a high-quality lead molecule for anticancer drugs.

The development of tailor-made tumor vaccines targeting specific tumor antigens is crucial for improving treatment accuracy. Tumor mRNA, as an emerging and promising vaccine modality, holds distinct technical edge in addressing this challenge, thus attracting considerable attention in cancer treatment. This article reviews the design and synthesis processes of tumor mRNA vaccines, highlighting the latest advances in antigen identification, optimization of RNA sequence and structure, and its delivery. Additionally, it discusses the challenges these vaccines face and outlines potential future directions for development.

Gefitinib, a first-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), exerts significant therapeutic efficacy in the treatment of non-small cell lung cancer (NSCLC) by selectively targeting mutant forms of EGFR. However, the development of acquired resistance significantly limits its long-term clinical benefits. Cell division cycle 20 (CDC20), a key regulator of cell cycle progression, has been implicated in the tumorigenesis and progression of various malignancies. Nevertheless, its role and underlying regulatory mechanisms in the acquisition of drug resistance in NSCLC remain largely unexplored. This study aimed to elucidate the molecular mechanisms by which CDC20 contributes to gefitinib resistance in NSCLC. Gefitinib-resistant cell lines, HCC827/GR (IC₅₀ 0.05 ± 0.01 μmol/L vs 36.24 ± 6.21 μmol/L) and PC9/GR (IC₅₀ 0.02 ± 0.01 μmol/L vs 25.36 ± 5.57 μmol/L), were established through stepwise drug induction, exhibiting markedly increased IC₅₀ values compared to their parental counterparts. Bioinformatics analysis revealed that the transcriptional level of CDC20 is significantly upregulated in lung cancer tissues and is associated with poor patient prognosis. Western blotting analysis confirmed elevated CDC20 protein levels in the resistant HCC827/GR and PC9/GR cells relative to the parental HCC827 and PC9 cells. To further investigate the functional role of CDC20 in NSCLC gefitinib resistance, CDC20 was knocked out using CRISPR/Cas9 technology. This genetic intervention significantly restored gefitinib sensitivity in resistant cells (IC₅₀ 37.08 ± 6.15 μmol/L vs 10.49 ± 1.83 μmol/L, 7.23 ± 1.55 μmol/L), while concurrently promoting apoptosis and inducing G₂/M phase cell cycle arrest. Conversely, CDC20 overexpression decreased drug sensitivity in parental cells and notably attenuated gefitinib-induced apoptosis and cell cycle arrest. Mechanistically, CDC20 depletion was found to inhibit activation of the PI3K/Akt/mTOR signaling pathway, upregulate pro-apoptotic proteins such as cleaved-Caspase 3 and Bax, and downregulate the anti-apoptotic protein Bcl-2. Collectively, these findings demonstrate that CDC20 mediates gefitinib resistance in NSCLC through modulation of the PI3K/Akt/mTOR signaling pathway, thereby identifying CDC20 as a potential therapeutic target for overcoming resistance to EGFR-targeted therapies.

The origin of life and its organizing principlesrepresent a long-standing hard problem in science. Because information, together with matter and energy, form the three cornerstones of our knowledge system and perhaps even the universe, we ask whether life can be defined from a purely mathematical and information science perspective. Here, we postulate a unified mathematic framework termed the Self-Information Theory of Life to define what life is and how it operates. Our theory states that all dynamic activity events in life—including DNA replication, RNA transcription, protein synthesis, cellular signaling, energy metabolism and even the brain’s high cognition—carry the hidden and intrinsic self-information based on their probability distributions using the ternary coding scheme. Specifically, the events with a higher probability distribution carry lower information content, whereas those lower probability events carry higher information content. The high-probability events represent the ground equilibrium state, whereas those low-probability states that deviate from this ground state signal positive surprisal or negative surprisal information, depending on the direction of the deviation. Thus, a given biochemical or biophysical step can be regarded as a self-information unit which generates the dynamic ternary information code explicitly tagged with a specific self-information value. A living organism is constructed by a set of self-information units organized in such a way to further produce their joint self-information probability distribution as the joint self-information groups. Through the feedback control mechanisms, these joint self-information groups form various closed loops to process both environmental inputs, to maintain internal homeostasis and to generate adaptive outputs, subsequently generating the chain of joint self-information flow. Those spatiotemporally ordered activities of each joint self-information group execute a specific given function such as gene replication, protein synthesis, energy metabolism, intracellular and neuronal signaling, learning and memory, intelligent and conscious behaviors, etc. The more advanced a life evolves (i.e. human), the larger numbers of the self-information units and joint self-information groups are, the richer self-information conscious levels and higher intelligence, thereby the lower its life entropy value is. Life and its activity defined by this purely mathematical and self-information principle surpass the traditional realm of matter and energy. This self-information theory of life may further provide a novel framework to design and build artificial life on earth or to explore and study extraterrestrial life in the Universe.

Biochemistry, as a fundamental course for science and engineering majors related to biology and chemistry, holds a significant position in the curriculum. The course team at Dalian Minzu University is committed to teaching innovation, adopting the outcome-based education (OBE) concept for teaching design and incorporating ideological and political elements, in order to achieve the dual goals of knowledge transmission and value guidance. The team has established a three-dimensional teaching goal of "knowledge, morality, and ability", covering "consolidating core knowledge, cultivating moral sentiment, and enhancing innovation ability". Through a multi-dimensional integrated teaching method of "three integrations and five combinations", multiple rounds of teaching practice have been carried out in the applied chemistry major using "classification and specificity of enzyme" as an example. The output of teaching results and survey questionnaires show that students highly recognize the teaching design and its "process-based learning" evaluation method, fully reflecting the student-centered teaching idea. Research has shown that OBE design combined with ideological and political elements can effectively promote students’ knowledge acquisition, moral growth, and innovation ability improvement in the course of Biochemistry. This teaching design not only helps students construct correct worldviews, outlooks on life, and values, but also significantly enhances their innovative thinking and practical abilities. This teaching design can not only effectively improve the teaching quality of the course, but also provide new perspectives and ideas for the teaching design of Biochemistry, realizing the organic integration of professional knowledge imparting and ideological and political education, and has certain innovation and practical significance.

Lactylation modification is a new type of protein post-translational modification, which mediates the covalent binding of lactic acid groups to lysine residues through amide bonds, thus changing protein function and intracellular signal transduction process. Lactylation modifications can be broadly categorized into two types: histone lactylation and non-histone lactylation, both of which are dynamically and precisely regulated by the "Writer-Eraser" enzyme system. Among them, non-histone lactylation, mainly regulated by enzymes such as AARS1 and SIRT3, plays an important role in the progression of many diseases, including tumor metabolic reprogramming, ROS stress and signal pathway regulation. Especially in tumors, non-histone lactylation is closely related to tumor proliferation, immune escape and drug resistance. Therefore, an in-depth study of the role of non-histone lactylation in the progression of tumors is expected to provide new targets and strategies for the accurate diagnosis and treatment of tumors. It is noteworthy that in the context of non-histone lactylation modification, the interference effect of acetylation modification cannot be ignored. Lactylation and acetylation share similar "writer" and "eraser" enzymes and exhibit overlapping modification sites, suggesting the possibility of functional crosstalk between the two. Due to the current lack of specific editing tools targeting lysine lactylation, it remains challenging to definitively determine whether lactylation plays a predominant regulatory role. This article reviews the research progress of non-histone lactylation in tumors in recent years.

The incidence of gastrointestinal tumors is increasing year by year and it has become a worldwide health problem with high incidence and poor prognosis. Ferroptosis, a new type of cell death, is mainly caused by abnormal intracellular iron metabolism leading to excess iron, which results in high production of intracellular reactive oxygen species (ROS) and accumulation of lipid peroxides. With the gradual deepening of the study of ferroptosis, it is found that ferroptosis can sensitize gastrointestinal tumor cells to drug therapy, so as to achieve a better therapeutic effect. Therefore, ferroptosis has attracted widespread attention in the field of treatment of gastrointestinal tumors in recent years. Traditional Chinese medicines have a long history and have been widely utilized in the treatment of cancer due to its low cost and fewer adverse effects. More and more studies have found that traditional Chinese medicine can induce ferroptosis in gastrointestinal tumors and thus inhibit tumor cell growth and metastasis. Here we first introduce the theory of ferroptosis, then further present traditional Chinese medicine monomers that induce ferroptosis in gastrointestinal tumor cells through modulating iron metabolism, inhibiting ferroptosis related antioxidant system and regulating nuclear factor erythroid 2-related factor 2 (NRF2), thereby inhibiting gastrointestinal tumorigenesis and progression. Moreover, we expound the research of anti-gastrointestinal tumor using traditional Chinese medicine and traditional Chinese medicine monomer combined with chemotherapy drugs to provide a new way of thinking for the treatment of digestive tract tumors.

The ubiquinol-cytochrome c reductase complex chaperone (BCS1L), a key regulator of the mitochondrial complex III assembly, plays a critical role in mitochondrial genetic disorders and prostate cancer-related fatigue, but its functional mechanisms in colon adenocarcinoma (COAD) remained unclear. This study systematically investigated the clinical significance and molecular mechanisms of BCS1L in COAD through integrated multi-omics analysis and functional experiments. Bioinformatics analysis of public databases revealed that BCS1L expression was significantly up-regulated in colon cancer tissues (P<0.05), with high expression correlating with poor prognosis and patient age. Functional enrichment analysis demonstrated a correlation of high BCS1L expression groups with organic anion/carboxylic acid transport pathways. The immune cell infiltration analysis revealed an increased proportion of regulatory T cells (Treg) cells in the high BCS1L expression group. The mutation profile results showed that the mutation frequencies of genes such as PCLO, ABCA13, LRP1B, FAT3, HYDIN, and SOX9 were significantly higher in the high BCS1L expression group (all P<0.05). Experimental validation confirmed that BCS1L knockdown significantly inhibited cell proliferation, clonogenicity, and migration in COAD (all P<0.05). Collectively, these findings demonstrate that BCS1L plays a pivotal role in promoting cell proliferation and migration by modulating transport pathways and facilitating the formation of an immunosuppressive tumor microenvironment in COAD.

Ribonucleotide reductase regulatory subunit M2 (RRM2), a key cell cycle regulatory gene, has been implicated in inducing cellular senescence, cell cycle arrest, or cell death across various cancers. Although previous studies have demonstrated the significant role of RRM2 in cancer cells, the specific mechanism by which RRM2 modulates ferroptosis via sorafenib (sorafenib, Sor) remains inadequately elucidated. This study aimed to investigate the mechanism underlying RRM2-mediated, sorafenib-induced ferroptosis in hepatocellular carcinoma (HCC) cells. Different concentrations of sorafenib (0, 10, 15, 20, 25, 30 μmol/L) were used to treat human hepatoblastoma cells (HepG2) and human hepatobiliary carcinoma cells (Huh7), and then the inhibitory effect of sorafenib on the proliferation of HepG2 cells and Huh7 cells was detected by CCK8 method. The optimal concentration for establishing HepG2 and Huh7 cell models was determined, including SOR group (transfected with empty plasmid combined with 11 μmol/L sorafenib intervention cells), OE-RRM2 group (RRM2 in overexpressed cells), OE-RRM2 sor group (transfected with RRM2 overexpression vector combined with 11 μmol/L sorafenib intervention cells), SINC group (transfected with empty vector as knockdown control), and SIRRM2-1, SIRRM2-2, and SIRRM2-3 group (RRM2 in knockdown cells). CCK-8 and plate cloning assay results indicated that RRM2 knockdown led to reduced cell proliferation, while RRM2 overexpression had the opposite effect (P＜0.05); Glutathione (GSH) assay results showed that RRM2 knockdown led to reduced cellular GSH levels, while RRM2 overexpression had the opposite effect (P＜0.05); Intracellular reactive oxygen species (ROS) assay results showed that RRM2 knockdown led to increased cellular ROS levels, while RRM2 overexpression had the opposite effect (P＜0.05); Mitochondrial membrane potential (MMP) assay results showed that RRM2 knockdown led to increased cellular MMP, while RRM2 overexpression had the opposite effect (P＜0.05); qRT-PCR and Western blot experiments showed that knocking down RRM2 led to decreased expression levels of glutathione peroxidase 4 (GPX4), glutathione synthetase (GSS), and hypoxia-inducible factor-1α (HIF-1α)/inducible nitric oxide synthase (iNOS)/vascular endothelial growth factor (VEGF) gene expression levels, while the RRM2 overexpression group showed the opposite results (P＜0.05). Immunofluorescence (IF) experiments showed that RRM2 knockdown led to a decrease of HIF-1α protein expression levels (P＜0.05). In summary, these findings suggest that sorafenib may activate RRM2 in liver cancer cells, thereby inhibiting ferroptosis. Furthermore, RRM2 may promote the malignant progression of HCC by activating the HIF-1α/iNOS/VEGF signaling pathway to suppress ferroptosis.