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.

Gene editing technology has become an important tool for biomedical research because of its high efficiency and precision in target gene localization and cleavage. The technology not only facilitates basic research on gene function, but also provides new strategies for gene therapy of hereditary diseases and genetic improvement of crops. With the integration of artificial intelligence (AI) technology, especially the application of machine learning algorithms, the design and execution of gene editing have become more intelligent. AI technology optimizes the design of sgRNAs through predictive analysis and pattern recognition, improving the specificity and efficiency of editing while reducing the risk of off-target effects. In addition, AI plays a key role in the parsing of large-scale genomic data, providing new perspectives for understanding complex biological processes and disease mechanisms. This paper reviews the research progress of data-driven gene editing technology in target precision, safety enhancement and personalized therapy, aiming to provide reference and inspiration for researchers in the field of gene editing technology and to promote the application and development of AI in gene editing technology.

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.

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.

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.

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.

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.

In the canonical model of gene transcription, transcription factors regulate the transcription of target genes by binding to double-stranded DNA (dsDNA) containing its specific consensus motifs. In contrast to dsDNA, G-quadruplexes are atypical nucleic acid secondary structures formed by guanine-rich sequences. G-quadruplex structures are involved in regulating various biological processes, including gene transcription, and have emerged as a significant research topic in the field of molecular biology. Recently, many research groups, including us, have revealed that G-quadruplex structures recruit transcription factors to bind to the promoters more efficiently than dsDNA, thereby enhancing target gene expression. However, there is currently a lack of comprehensive summaries and discussions regarding this atypical model of gene transcription regulation. In this review, we first elucidate the characteristics of G-quadruplex structures and the techniques used to identify these structures. G-quadruplexes can be classified into intramolecular and intermolecular types; intramolecular G-quadruplexes are further divided into parallel, antiparallel and hybrid types. The G-quadruplex structures can be determined using techniques such as circular dichroism, nuclear magnetic resonance spectroscopy and gel migration, among others. Then, we discuss the regulatory role of G-quadruplex in gene transcription. G-quadruplexes are mainly highly enriched in gene promoter. Early studies revealed that G-quadruplexes can inhibit gene transcription. However, lots of studies have recently proven that this structure has a new function of recruiting transcription factors to activate gene transcription. Finally, we summarize the classification of transcription factors with G-quadruplex binding activity, including transcription factors with C₂H₂ zinc fingers, forked head/winged helices, and p53 domains. The DNA-binding domain determines the interaction between transcription factors and G-quadruplex. Moreover, we propose future research directions in the field of G-quadruplex and transcription factors in regulating gene transcription. In conclusion, this review can provide important guidance for understanding the concept of G-quadruplex as a cis-acting element for transcriptional activation.

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.

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.

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.

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.

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.

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.

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.

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.

The incidence and mortality rate of lung cancer rank among the highest worldwide, severely endangering human health and life. Metformin, an anti-diabetes drug, has been shown to elicit anticancer activities in various tumors. However, its underlying mechanisms remain elusive. In this work, we explore the role of receptor-interacting protein 1 (RIP1) which plays a crucial role in the process of cell death, in metformin-induced anticancer activities in lung cancer. Metformin inhibits lung cancer cell proliferation in a dose-dependent manner and promotes apoptotic cell death, as evidenced by metformin-induced PARP and caspase cleavage. Furthermore, the pan-caspase inhibitor z-VAD-fmk reverses metformin-induced cell death. Western blot and qPCR results suggest that metformin markedly downregulates RIP1 expression without affecting its mRNA and ubiquitination levels (0 vs 80 mmol/L, 100% vs 20%, 100% vs 15%). Additionally, co-immunoprecipitation and immunofluorescence results reveal that metformin may suppress RIP1 expression in an Hsp70-dependent manner, as metformin promotes Hsp70 degradation, and Hsp70 endogenously interacts with RIP1. Subsequent CCK-8, flow cytometry, and Western blot analyses suggest that metformin decreases Hsp70/RIP1 expression through AMPK/PKA/GSK-3β axis. Consistently, results from a subcutaneous transplant tumor model indicate that metformin retards tumor growth without affecting mouse body weight. Collectively, these data highlight the part of RIP1 in metformin-induced anticancer activities in lung cancer in vitro and in vivo, providing novel strategy for lung cancer administration.

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.

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.

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.

Collagen is a matrix protein essential for maintaining the function of various tissues in animals. There are many types of collagen in vertebrates, and the function of each type of collagen in the body is closely related to its sequence and structure. In addition to the common Gly-X-Y amino acid sequence, there is also a special structure Gly-Gly-Y in the natural collagen. In order to explore its effect on collagen, this study constructed mutants containing Gly-Gly-Y at the levels of collagen polypeptides, long-chain collagen, and collagen polymers. The thermal stability of the mutant before and after mutation was characterized by circular dichroism scanning. At the same time, molecular dynamics simulation was used to calculate the hydrogen bonding probability and bending degree of collagen polypeptides, explaining the molecular mechanism of changes in collagen stability and flexibility. The results showed that the mutant containing the Gly-Gly structure would decrease the Tm value of the sample, but this effect would gradually weaken as the collagen triple helix region lengthened and the degree of protein self-assembly increased, and the reduced Tm values are 5 ℃, 3 ℃, and 1 ℃, respectively. At the same time, the simulation results show that the curvature of the polypeptide containing the Gly-Gly structure also increases to some extent, indicating that the structure near the mutation site has greater flexibility. This provides a new idea for the design of rubber materials with certain flexibility.

Potassium-calcium activates channel subfamily N member 3 (KCNN3/SK3/ KCa2.3) is involved in regulating cellular calcium signaling, muscle contraction and neurotransmitter release. Dysregulation of the KCNN3 channel is associated with the development of various tumors. We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma (STAD) as a prognostic target. By analyzing the Human Protein Atlas (HPA) database and The Cancer Genome Atlas (TCGA) database, we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD, and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD. In addition, we found that high expression of KCNN3 in STAD reduced the IC₅₀ of several drugs in STAD cells, suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD. To investigate the underlying biological mechanism, we identified a potential KCNN3 interaction factor, tumor necrosis factor receptor superfamily member 7 (CD27/TNFRSF7), which is expressed at low levels in STAD. RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels, and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm. Moreover, we confirmed the inhibitory effect of KCNN3 on the proliferation, migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments. Furthermore, GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport. Lastly, we verified through cell co-culture, RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells. Therefore, our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.

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 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.

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.

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.

Neuropathic pain (NP) is a type of chronic pain caused by damage or disease of the nervous system. It is mainly characterized by spontaneous pain, hyperalgesia, and allodynia, which seriously affect the quality of life of patients. The pathogenesis of NP is complex, involving abnormal regulation such as peripheral sensitization, central sensitization, ion channel changes, and glial cell activation. In recent years, the role of m6A in NP has attracted extensive attention. However, the research on the role of m⁶A modification in different diseases and pain models is still limited. Therefore, it is particularly important to clarify the role of m6A modification in different diseases and pain models. This article reviews the research progress on the role and mechanism of m⁶A methylation modification in the pathogenesis of NP in recent years, especially the role mechanism of the five classical m⁶A modification factors, METTL3, METTL14, FTO, ALKBH5 and YTHDF1, in mediating the formation of NP in different diseases and pain models, with the expectation of providing new insights and ideas for the drug development and prevention of NP from the perspective of m6A modification.