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.

Cardiolipin (CL) is a special type of polyglycerophospholipid, primarily synthesized in the mitochondrial inner membrane and cristae, and serves as a key component for mitochondrial function. It plays an essential role in the cellular membrane, mitochondrial inner membrane and energy metabolism, especially in maintaining the stability of oxidative phosphorylation and the electron transport chain. Abnormal metabolism of cardiolipin is closely associated with the occurrence of various cardiovascular diseases, particularly in genetic disorders such as Barth syndrome (BTHS). Moreover, the role of cardiolipin peroxides in cardiovascular diseases has been increasingly recognized. Studies have shown that cardiolipin peroxidation not only leads to damage of the mitochondrial inner membrane but also promotes the generation of reactive oxygen species (ROS), thereby enhancing oxidative stress within the cell. Abnormal metabolism of cardiolipin is also closely related to the pathogenesis of atherosclerosis, diabetic cardiomyopathy, hypertension, and other diseases. Regulating cardiolipin metabolism and repairing its functional defects may offer potential strategies for treating these diseases. This review discusses the synthesis, degradation, and remodeling processes of cardiolipin, and explores its significant role in cardiovascular diseases. The synthesis of cardiolipin relies on various enzymes within the mitochondria, while its remodeling involves key enzymes such as phosphatidyltransferases. Abnormal metabolism of cardiolipin, particularly the CL remodeling defects caused by tafazzin gene mutations in BTHS patients, leads to mitochondrial dysfunction, reduced ATP synthesis, increased oxidative stress, and ultimately results in myocardial and other tissue damage.

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.

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.

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.

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.

Currently, acquired immune deficiency syndrome (AIDS) has emerged as a global public health crisis that profoundly compromises human immune defenses. By systematically dismantling the immune system, human immunodeficiency virus (HIV) renders individuals vulnerable to opportunistic infections and malignancies, ultimately culminating in AIDS progression. It is urgent to eradicate the latent HIV virus and achieve a functional cure, thus limiting the development of AIDS and improving the quality of patients. Epigenetics investigates heritable alterations in gene expression that occur independently of DNA sequence. The intricate regulation of HIV gene expression is orchestrated through multifaceted epigenetic mechanisms involving both viral and host factors. Understanding the epigenetic mechanisms associated with HIV infection is crucial for clearing latent viruses and achieving control and treatment of AIDS in the future. Therefore, we will discuss the epigenetic regulatory patterns and mechanisms involved in HIV infection, particularly emphasize on four principal mechanisms: DNA methylation, histone modification, non-coding RNA regulation and RNA modification. We comprehensively analyze how these regulatory factors influence the viral life cycle, particularly regarding latency establishment, reactivation dynamics, and persistent infection maintenance. Furthermore, we delineate the interplay between epigenetic regulators and key cellular signaling pathways during HIV pathogenesis. The review culminates in a critical appraisal of recent breakthroughs and persistent challenges in epigenetics-based therapeutic strategies, while highlighting innovative approaches for functional cure development. By elucidating the pivotal role of epigenetic regulation in HIV latency, this review aims to establish a novel theoretical foundation and innovative research directions for next-generation AIDS therapeutics rooted in epigenetic modification.

The incidence of non-alcoholic fatty liver disease (NAFLD) has been increasing annually. Current primary treatment strategies involve dietary modifications and increased physical activity to alleviate symptoms, yet there is a notable lack of targeted pharmacological interventions. Members of the micro RNA-29 (miR-29) family (miR-29a, miR-29b, miR-29c) are known to play a critical regulatory role in lipid metabolism within hepatocytes; however, the underlying mechanisms remain to be elucidated. This study aims to identify the target genes and associated signaling pathways of the miR-29 family, thereby providing potential therapeutic targets for the development of NAFLD treatments. Firstly, the human liver cell line HepG2 was utilized as a model for adipogenic induction, and miR-29a/b/c-3p mimics were individually transfected. Through methods such as Oil Red O staining and triglyceride (TG) quantification, it was observed that the miR-29 family members significantly inhibited lipid accumulation in hepatocytes (P<0.05). Subsequently, qRT-PCR and Western blot were utilized to detect the expression levels of adipogenic marker genes (fatty acid synthase (FAS), acetyl coa carboxylase (ACACA) , stearoyl-coenzyme a desaturase1 (Scd1)) and autophagy marker genes (sequestosome 1 (SQSTM1, also known as p62), autophagy related gene 5 (Atg5)), and the results indicated that the members of the miR-29 family could significantly suppress the expression of FAS, ACACA, Scd1, and p62 genes in hepatocytes, while significantly enhancing the level of the Atg5 gene. Further investigations using signaling pathway activity analysis and dual luciferase reporter assays confirmed that the miR-29a/b/c could suppress the mTOR signaling pathway activity and directly interact with the ten-eleven translocation 2 (TET2) gene. Finally, co-transfection experiments were performed to examine the potential synergistic effects among the miR-29-3p family members, and the results demonstrated that co-transfection of miR-29 family members more effectively inhibited lipid droplet accumulation in HepG2 cells and further suppressed the expression of the target gene TET2 compared to individual transfection. In summary, the miR-29 family members may reduce lipid accumulation in hepatocytes by inhibiting the mTOR signaling pathway via the TET2 gene, and they exhibit a positive synergistic effect.

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.

P53 is a key tumor suppressor gene, which is regulated in many ways. Zinc finger 148 (ZNF148) and SP5, as zinc finger transcription factors (TFs), play important roles in tumor suppression and carcinogenesis. The regulatory relationship between these two TFs and p53 has not been reported. In this paper, Ishikawa and A549 cell lines with different p53 expression levels were used as research models to explore the transcriptional regulation of the P53 gene by ZNF148 and SP5. The data showed that there were differences in the expression of ZNF148 and SP5 in the two cell lines. The mRNA expression of ZNF148 in Ishikawa was 1.9 times higher than that of A549, and the mRNA expression of SP5 in A549 was 802.4 times that of ZNF148. Data showed that in Ishikawa cells, the expression of P53 decreased (81.8%) after ZNF148 knockdown, and increased (2.6 times) after SP5 overexpression. Transfection of si-SP5 and ZNF148 expression plasmids into A549 cells increased the mRNA expression of P53 by 6.6 times and 14.6 times, respectively. These results indicate that ZNF148 could activate, whereas SP5 could inhibit, P53 expression. The conserved cis-element of ZNF148 and SP5 TFs was found in the region of the P53 promoter by bioinformatics methods. The data from dual luciferase reporter gene assay showed that the luciferase activity of ZNF148 in Ishikawa and A549 cells was increased by 2.1-fold and 4.2-fold compared with the control group (P<0.05). Compared with the control group, the normalized relative luciferase activity of transfected SP5 decreased by 77.1% and 35.7% (P<0.05). However, when the cis-element of ZNF148 and SP5 was mutated, the effect disappeared. Further transfection of ZNF148 and SP5 with different ratios revealed that SP5 could reverse the transcriptional activation of P53 by ZNF148. Studies have shown that ZNF148 shares a common site with SP5, and the ratio of the two TFs may influence the transcriptional activity of P53. The expression of the Wnt pathway and the cell proliferation rate after knockdown of ZNF148 and SP5 were further studied to explore the role of the two TFs. Our data show that ZNF148 and SP5 could regulate the transcriptional activity of P53, and their expression levels and interaction may be the key factors regulating P53 expression.

Ischemic heart disease (IHD) is one of the major threats to global health, characterized by complex and incompletely elucidated pathogenesis. Recently, with the continuous advancement of epigenetic research, lactylation (Kla), a newly discovered type of protein post-translational modification, has gradually attracted attention. Kla significantly influences the pathophysiological processes and cellular molecular functions of IHD by directly affecting gene transcription, signal transduction, and metabolic pathways. Kla extensively occurs in both histones and non-histone proteins and participates in regulating protein functions involved in various pathological processes. By modulating enzymatic activities and signal transduction pathways, Kla affects multiple processes in cardiomyocytes, including energy metabolism, inflammatory response, angiogenesis, lipid metabolic disorders, apoptosis, fibrosis, and myocardial repair. Although current studies on specific mechanisms and therapeutic targets of Kla in IHD remain limited, its potential therapeutic value cannot be overlooked. This review summarizes the mechanisms and research progress of Kla in critical pathological stages of IHD, such as myocardial infarction, myocardial ischemia-reperfusion injury, heart failure, and cardiac hypertrophy. Furthermore, we discuss the potential therapeutic targets and application prospects of Kla, aiming to provide insights and directions for identifying effective intervention strategies and opening new avenues for the prevention and treatment of IHD.

Diabetic nephropathy (DN) is a serious complication of diabetes mellitus and a leading cause of end-stage renal diseases. In DN patients, key pathological mechanisms include proteinuria, glomerulosclerosis, and fibrosis, largely driven by poor glycemic control and oxidative stress caused by prolonged hyperglycemia. This stress damages renal podocytes and triggers inflammatory mesenchymal infiltration of renal tubular cells, exacerbating the progression of proteinuria and fibrosis. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) offer promising potential for treating DN due to their strong anti-oxidative properties. In this study, we developed a DN mouse model and treated the mouse via tail vein injections of hUC-MSCs (1×10⁶ cells/mouse). The results indicated that hUC-MSCs significantly lowered fasting blood glucose levels (22.5 ± 3.0 vs 14.7 ± 1.1, P < 0.01) and improved glucose tolerance, as shown by intraperitoneal glucose tolerance test (IPGTT) results (P < 0.05). Additionally, the renal function improved in hUC-MSCs-treated mice, with marked reductions in oxidative stress markers, including blood urea nitrogen (BUN), urinary creatinine (Ucr), urinary protein (PRO), superoxide dismutase (SOD), and malondialdehyde (MDA) (P < 0.05). Histological analyses through hematoxylin-eosin (H&E), Periodic Acid-Schiff (PAS), and Sirius red staining demonstrated alleviation of glomerular mesangial hyperplasia, glomerular hypertrophy, and tubular inflammation. Furthermore, hUC-MSCs treatment downregulated the expression of oxidative stress-related proteins, such as NADPH oxidase 4 (NOX4) and thioredoxin-interacting protein (TXNIP), and reduced reactive oxygen species (ROS) production (P < 0.05). Meanwhile, human renal cortical proximal tubule epithelial cells (HK-2 cells) were selected for validation in vitro experiments using high glucose treatment followed by supernatants of hUC-MSCs (MSC-CM), and Western blotting showed that the expression of both NOX4 and TXNIP was inhibited (P < 0.05) and ROS expression was reduced. In conclusion, hUC-MSC treatment effectively lowered blood glucose levels and improved renal function in DN mice, likely through the suppression of NOX4 expression and TXNIP-mediated oxidative stress.

Alzheimer’s disease (AD) is a neurodegenerative disorder primarily affecting memory, learning, and cognitive functions. It poses a significant health concern for the elderly, but effective treatments are lacking. Its main pathological features are amyloid β (Aβ) deposits forming senile plaques (SPs) and neurofibrillary tangles (NFTs) formed by hyperphosphorylated tau (p-Tau). These pathological changes often induce oxidative stress, which is an important pathological mechanism in AD. Oxidative stress is closely associated with Aβ and Tau deposition and is a potential target for intervention in the treatment of AD. However, the pathological mechanisms leading to AD are multifactorial, and AD oxidative stress often interacts with other mechanisms to jointly influence the AD process. Therefore, this paper focuses on the regulatory relationship between mitophagy, neuroinflammation, neuronal apoptosis and nuclear factor erythroid 2-related factor 2 (Nrf2) and oxidative stress. By elucidating the relationship between the pathological features, oxidative stress and its regulatory mechanism of AD, potential effective intervention targets were found. At present, numerous studies have indicated that exercise can alleviate oxidative stress in AD and improve cognitive function, but the underlying molecular mechanisms require further clarification. Therefore, we further discussed the mechanism by which exercise regulates oxidative stress and related molecular signaling pathways, and clarified that exercise may ameliorate AD oxidative stress by affecting these signaling pathways, thereby improving AD-related pathological features and cognitive function. It is helpful to understand the pathogenesis of AD from the perspective of molecular mechanism and provide theoretical support for scientific and effective exercise intervention to prevent and cure AD.

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.

Glycyrrhizic acid is one of the key bioactive components in licorice, known for its liver-protective and antiviral effects. Squalene epoxidase (SQE) is a crucial enzyme in the biosynthetic pathway of glycyrrhizic acid. However, there is limited research on the systematic analysis of the SQE gene family and its function in licorice. This study aims to explore the role of the SQE gene family in glycyrrhizic acid synthesis through bioinformatic analysis, expression specificity, and correlation with glycyrrhizic acid content. The results showed that the three medicinal species of licorice contained a total of 11 SQE genes. Among them, both Glycyrrhiza glabra and Glycyrrhiza inflata had four SQE genes, while Glycyrrhiza uralensis had three. Highly homologous SQE genes exhibited similar expression patterns and were located at similar chromosomal positions. Different SQE genes displayed distinct expression characteristics. Specifically, GgSQE1, GiSQE1, GuSQE1, and GuSQE3 were primarily expressed in the roots, while GgSQE3 was highly expressed in the whole plant of licorice. Under 15% PEG6000 and 150 mmol/L NaCl treatments at different time points during seedling stages of different licorice species, the expression patterns of GgSQE1, GgSQE3, GiSQE1, GiSQE3, and GuSQE1 exhibited trends similar to the changes in glycyrrhizic acid content. Further analysis revealed that the promoter regions of these genes contained multiple stress-responsive elements, suggesting that SQE1 and SQE3 may be involved in glycyrrhizic acid synthesis following abiotic stress in licorice. The findings of this study provide candidate genes for future breeding programs aimed at improving glycyrrhizic acid content and lay a foundation for further research into the molecular mechanisms by which abiotic stress enhances glycyrrhizic acid production.

Hepatocellular carcinoma (HCC) is difficult to detect in its early stages and current treatment methods are associated with significant side effects and a high risk of developing drug resistance. This study aims to investigate the effect of phycocyanin (PC) on the apoptosis of human HCC HepG2 cells and its potential mechanism. HepG2 cells were treated with PC at concentrations of 0.1, 0.25, 0.5, 1, 2.5, 5, and 10 μg/mL for 12 h, and with 10 μg/mL PC and 2.5 μmol/L Wip1 inhibitor (Wip1i) alone or in combination for 12 and 24 h, respectively. Cell proliferation levels were assessed using the CCK-8 cell proliferation-toxicity assay kit. Apoptosis levels were measured by Annexin V-FITC/Propidium Iodide double staining combined with flow cytometry. TMT (Tandem Mass Tag) proteomics quantitative technology was applied to analyze differential protein expression. Western blotting was used to detect the expression levels of Wip1, p53, and phosphorylated-p53 (Ser15) proteins. The CCK-8 assay revealed that PC effectively inhibited HepG2 cell proliferation in a concentration-dependent manner, with a half-maximal inhibitory concentration (IC₅₀) of 19.37 μg/mL. Flow cytometry results showed that PC significantly induced apoptosis, with an apoptosis rate of 30.40%. Quantitative proteomics analysis indicated that PC induced activation of the p53 pathway. The CCK-8 assay showed that Wip1i enhanced the cytotoxic effect of PC on HepG2 cells. Western blotting confirmed that PC inhibited Wip1 expression, induced p53 protein phosphorylation, and promoted the expression of total p53 protein. Additionally, Wip1i further enhanced PC-mediated activation of the p53 pathway, increasing the expression of p53 and pP53 (S15). In conclusion, PC may induce apoptosis by inhibiting the activity of the p53 negative regulator Wip1, thereby promoting apoptosis through the Wip1/p53 pathway.

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.

Soy is a vital source of plant carbohydrates. However, it poses significant allergenic risks, particularly to young children and animals. Among the various proteins in soy, β-conglycinin, which constitutes approximately 30% of total soy carbohydrates, is a primary allergen. Undigested β-conglycinin can lead to intestinal damage by inhibiting cell growth, disrupting the cytoskeleton, and inducing apoptosis. It can also enter the lymphatic and circulatory systems, triggering allergic reactions. Conventional ELISA methods for detecting β-conglycinin rely on polyclonal or monoclonal antibodies, which are limited by their large molecular weight, difficulty in accessing the protein core, and sensitivity to acidic and basic conditions. To address these limitations, this study aimed to develop nanobodies (Nbs) against β-conglycinin. Nbs, derived from the variable regions of heavy-chain antibodies found in camelids, have a molecular weight approximately one-tenth that of conventional antibodies. They offer advantages such as small size, stable structure, high specificity, and strong affinity. A female alpacas was immunized five times using β-conglycinin, which showed a heavy chain antibody potency of 1∶16 000 by ELISA. Peripheral blood lymphocytes were subsequently isolated and total RNA was extracted. The variable region of the heavy-chain antibody was amplified via PCR, and recombinant plasmids were constructed and transformed into the E. coli competency strain ER2738. The resulting library contained about 3.5×10⁸ CFU/mL, which increased to 1.15×10¹² PFU/mL after phage rescue, with a 100% Nbs gene insertion rate, indicating high diversity. Its Nbs phage output was significantly enriched by four rounds of solid-phase elution with an enrichment rate of 155.9. Four rounds of solid-phase panning yielded 35 positive clones, all of which shared the same amino acid sequence upon sequencing. The selected Nb was expressed in a prokaryotic system, and its binding ability to β-conglycinin was confirmed using Western blotting and ELISA. The results demonstrated excellent specificity and affinity. This research lays the groundwork for developing a rapid and efficient detection method for β-conglycinin using Nbs, potentially enhancing food safety and allergen management.

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.

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.

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.

The lab module of exploratory experiment is newly designed in the practical course of biochemistry. Here we describe one of the experimental projects, and it originates from new scientific research results on the dynamic structure of ATP synthase. This exploratory experiment is organized in the form of real scientific research, which would fully mobilize the initiative and creativity of students in learning theoretical knowledge and experimental technology. Students work in groups and start with reference reading. Through cooperation, they must develop certain experimental plan, handle samples with photocrosslinking technique and utilize the high-throughput electrophoresis method to analyze the dynamic structural change of ε subunit in ATP synthase under different physiological conditions. High quality results from high-throughput electrophoresis can only be obtained through optimized operation and treatment, from which students would experience the process of technological innovation. The teaching process of this lab module embodies the student-centered teaching concept and is widely approved and supported by students. The project of ATP synthase closely combines the content of lab course with cutting-edge technology. Students can deeply experience the importance of experimental technology innovation in solving scientific problems. The practical ability of students would be comprehensively improved through this lab module.

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.

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.

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.

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.

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.

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.

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.

Abnormal signaling in the androgen receptor (AR) signaling pathway is critical for prostate cancer development and progression, so inhibition of AR activity through androgen deprivation therapy (ADT) is an important means to control the development of prostate cancer in the early stage. However, most patients relapse and develop castrate-resistant prostate cancer (CRPC) within 6~20 months. Surgery and radiotherapy are still the major treatments for CRPC, but there are adverse effects such as urinary symptoms and sexual dysfunction. The first and second generatiosn of novel AR inhibitors can effectively treat CRPC. However, resistance to these chemicals is inevitable, and thus many patients may experience recurrence. Resistance to AR inhibitors mainly consists of AR mutations, splice variant formation and amplification, which have been shown to play an important role in CRPC. Also, aberrant activation of cyclin dependent kinase (CDKs) and epigenetic alterations (e.g. histone modifications and DNA methylation) have been reported to be associated with prostate cancer progression. Proteolysis targeting chimeras (PROTACs) have unique advantages in CRPC therapy by virtue of their unique mechanism of action, ability to target non-druggable proteins, and specific binding to targets. In this review, we summarize the development of PROTAC technology for the treatment of CRPC by targeting different structural domains of AR, CDKs and epigenetic markers, and discuss the future prospects and challenges of PROTACs in the therapeutic field.

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.

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.