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.

The emergence of CRISPR/Cas system has greatly promoted the progress in the field of gene editing, especially CRISPR/Cas9 system, which has become a core tool in biomedical research. Long noncoding RNAs (lncRNAs) play a key role in gene regulation, cell differentiation and the development of a variety of diseases. Especially in cancer research, lncRNAs have important application prospects as cancer biomarkers and therapeutic targets. However, lncRNAs are generally characterized by low abundance and poor conservation, which limits the study of their functions by traditional means. CRISPR/Cas9 technology provides an efficient, flexible and accurate tool for lncRNA research, which significantly accelerates the progress in this field. This paper first reviews the basic principles of CRISPR/Cas9 system and its wide applications in gene editing, including CRISPR knockout, knock-in, interference, activation and other functional systems. These technologies can not only screen key lncRNAs in specific biological processes, but also be used for gene function research to explore their roles in diseases. This article focuses on the analysis of CRISPR/Cas9 technology in the study of lncRNA functions, regulatory mechanisms, and its key applications in tumor research. In addition, the article also summarizes the methods of genome-wide screening by CRISPR/Cas9 to identify functional lncRNAs, and discusses the roles of these lncRNAs in cancer cell proliferation, migration, invasion and drug resistance. CRISPR/Cas9 knockout system can efficiently knock down lncRNA genes and reveal their specific functions in gene regulation. At the same time, CRISPR activation and interference technology provide a new idea for the research of noncoding genes, and further explore its clinical application in cancer and other diseases by regulating the expression level of lncRNAs. The article also discusses the potential of CRISPR technology in future lncRNA research, especially the progress in solving technical problems such as genome complexity, targeting efficiency and off-target effects. As mentioned in the review, CRISPR/Cas9 technology not only provides a powerful tool for studying lncRNAs, but also provides new ideas and opportunities for developing new means of cancer diagnosis and treatment in the future.

Ferroptosis is a new form of programmed cell death with iron-dependent lipid peroxidation as its core. A variety of metabolites are involved in the regulation of ferroptosis, among which lipid metabolism plays an important role. The most classic lipid metabolism-mediated ferroptosis mechanism is that phospholipids containing polyunsaturated fatty acyl chain (PUFA-PLs) located on biological membranes undergo super-threshold peroxidation, which leads to the destruction of membrane structure and function. In addition, special lipids containing polyunsaturated fatty acid (PUFA), such asphospholipid withdiacyl-PUFA tails (PL-PUFA₂), polyunsaturated ether phospholipid (PUFA-ePL), cholesterol ester containing polyunsaturated fatty acyl chain (PUFA-CE) have also been found to be involved in the ferroptosis process by providing PUFA for peroxidation. Lipid droplets was also found to regulate the sensitivity of ferroptosis through storing and releasing PUFA. Intermediates and derivatives of cholesterol metabolism are mainly involved in the negative regulation of ferroptosis, whereas different classes of sphingolipids were reported to have inconsistent regulatory directions for ferroptosis. A large number of previous studies have confirmed that ferroptosis is closely related to the metastasis and drug resistance of gastrointestinal tumors, therefore, we further summarized the lipid metabolism mechanisms related to ferroptosis resistance in gastrointestinal tumor cells, such as weakening the anabolism and peroxidation processes of PUFA-PLs, enhancing the ferroptosis defense system and so on. At the same time, we elaborated on the relationship between cholesterol metabolism, lipid drop metabolism, sphingolipid metabolism, and ferroptosis resistance in gastrointestinal tumors. Targeting these specific lipids, metabolic enzymes, and pathways to regulate ferroptosis has important clinical potential value. It is expected to provide new ideas for finding new diagnostic and prognostic markers, therapeutic drugs, and reversing chemotherapy resistance for gastrointestinal tumors.

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.

Aging is the result of the accumulation of various molecular and cellular damages over time, encompassing 12 characteristic hallmarks divided into three categories. These include primary hallmarks such as genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, and disabled macroautophagy; antagonistic hallmarks like deregulated nutrient sensing, mitochondrial dysfunction, and cellular senescence; and integrative hallmarks including stem cell exhaustion, altered intercellular communication, chronic inflammation and dysbiosis. Therefore, investigating cellular signaling factors within a single pathway is insufficient to comprehensively understand the complex mechanisms underlying aging. Casein kinase II (CK2), one of the earliest identified protein kinases, is capable of phosphorylating serine/threonine/tyrosine residues on hundreds of substrates. It exhibits highly constitutive expression and activity, and is extensively involved in the regulation of cellular processes such as proliferation, differentiation, apoptosis, stress response, metabolism, and immune function. CK2 plays a unique role in coordinating the cross-talk and integration of various signaling pathways, which is crucial for maintaining cell survival and homeostasis. Recent studies have revealed that CK2 exhibits perturbations in both expression levels and enzymatic activity during aging process, with notable heterogeneity observed across different animals, tissues and cellular models. Overall, the downregulation of CK2 expression not only promotes the development of primary hallmarks but also alleviates antagonistic hallmarks and ameliorates integrative hallmarks of aging, demonstrating dual effect and interconnected mechanisms. Notably, aberrant activation in CK2 expression and activity are associated with various aging-related diseases, including cancer, cardiovascular diseases, chronic metabolic disorders, neurodegenerative diseases and skeletal degenerative conditions. Therefore, maintaining CK2 homeostasis may represent an effective strategy for delaying aging. This review summarizes the latest advances in CK2 and aging research, providing not only deeper insights into the common mechanisms underlying aging and aging-related diseases, but also a theoretical foundation for identifying potential targets for the early prevention of aging-related diseases.

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.

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.

Liquid-liquid phase separation (LLPS) is the process where intracellular biomolecules rapidly form reversible high concentrations of liquid-phase condensates, resulting in the compartmentalization and the formation of intracellular membraneless organelles. LLPS is involved in various biological processes and pathologic processes, such as neurodegenerative diseases and cancers. Long noncoding RNAs (lncRNAs) have been revealed to be closely related to phase separation. It has become a research hotspot in life science recently, because it provides new insight into the mechanism of lncRNAs. In this review, we focuses on the effect of lncRNA SLERT on the phase separation of nucleolar fibrillar center/dense fibrillar component (FC/DFC) by interacting with DDX21 protein as a molecular chaperone; LINC00657 (NORAD) forms NP bodies with PUM proteins, which drives PUM protein liquid droplets and inhibits its activity, and promotes genomic stability; DilncRNA regulates DNA damage response small RNAs (DDRNAs) and LLPS of p53 binding protein 1 (53BP1) in response to DNA damage, and lncRNA LINP1 phase separation droplets bind to Ku protein to promote DNA damage repair; LncRNA SNHG9, MELTF-AS1 and MALR drive LATS1, YBX1 and ILF3 protein LLPS respectively to promote cancer, while GIRGL and lncFASA act as tumor suppressor genes in cancer development through regulating the phase separation of CAPRIN1 and PRDX1 respectively; LncRNA XIST drives X chromosome inactivation by LLPS. In a word, we summarize the latest research progress about the functional roles of lncRNAs in FC/DFC nucleolus, genomic stability and DNA damage and repair, cancer and X-chromosome inactivation through regulating LLPS. This paper shows that lncRNAs can participate in multiple pathophysiological processes by regulating LLPS, which is expected to provide a new direction for the treatment of LLPS-mediated diseases.

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.

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 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.

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.

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.

Circular RNA (circRNA) is a single-stranded RNA with a covalently closed loop structure, which has a more stable structure and lower immunogenicity than linear RNA. Many studies have shown that circRNA has characteristics such as conservation, stability, and tissue specificity, and can act as a microRNA (miRNA) sponge to interact with proteins and translation templates, and regulate biological functions such as gene expression and signal transduction. Based on the characteristics and various biological functions of circRNA, some endogenous circRNA plays an important regulatory role in the occurrence and development of tumors and has the potential to be used as biomarkers and therapeutic targets. In addition, mRNA drugs have limitations such as instability, easy degradation, low translation efficiency, and immunogenicity in practical applications. Engineered translatable exogenous circRNA can solve some of the limitations of linear mRNA application and become a new type of potential efficient drug. In this paper, we introduce the circRNA biogenesis mechanisms, specific biological functions, and the current status of diagnosis and treatment applications in tumors. This includes the diagnostic application of endogenous circRNA in tumors, the design and synthesis strategies of exogenous circRNA, and the current progress in the design and application of engineered circRNA vaccines using their stable and efficient protein expression functions in the treatment of tumors. Finally, we discuss the current clinical diagnostic application problems of circRNA, the challenges of exogenous circRNA therapeutic applications, and the prospects of the field.

Metabolic reprogramming is a crucial feature of cancer. Among various metabolic processes, oxidative phosphorylation (OXPHOS) metabolism serves as the primary biochemical pathway for energy production and significantly influences tumorigenesis and tumor development. Therefore, this study aims to explore the impact of the metabolic characteristics of lung squamous cell carcinoma (LUSC) on its malignant properties. By analyzing the single-cell transcriptome sequencing data of LUSC, lung adenocarcinoma, and normal lung tissues from the gene expression profile database, it was found that the OXPHOS signaling pathway and molecules related to ATP synthesis were remarkably enriched in LUSC tissues. Based on the OXPHOS signal intensity score, LUSC cells were classified into OXPHOS^high and OXPHOS^low groups. Bioinformatics analysis revealed that 126 transcription factors were highly expressed in both LUSC tumor tissues and the OXPHOS^high group. Notably, the expression levels of cancer stem cell-related signals (such as SOX2, SOX9, POU2F1, CDX1, ARID3A, EZH2, and KLF5) were significantly enhanced in the OXPHOS^high cell subset (P<0.05). Treatment of LUSC cells with an OXPHOS antagonist significantly inhibited the sphere formation rate of H520 and SKMES-1 cells, which is a key characteristic of cancer stemness (P <0.05). Analysis of cell-cell communication data from the LUSC single-cell transcriptome indicated that the signal emission and reception in OXPHOS^high cells were stronger than those in OXPHOS^low cells. Moreover, fibroblasts showed significant interaction with OXPHOS^high-LUSC cells. Co-culture of LUSC cell lines H520 and SKMES-1 with human lung fibroblast-like cell lines significantly increased the sphere formation rate of tumor cells (P <0.05). Additionally, the levels of ATP, NADH-active enzymes, and reactive oxygen species (ROS) in co-cultured H520 and SKMES-1 cells were significantly elevated (P <0.05). The results of this study confirm that the OXPHOS pathway is a key signal involved in LUSC metabolism, which is closely associated with the characteristics of cancer stem cells and the regulatory signals of fibroblast interactions. This finding holds promises for providing novel strategies for tumor treatment.

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.

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.

The ocean plays a critical role in the global carbon cycle, and base on the "dual carbon" goals, ocean carbon sinks have received widespread attention. Shellfish aquaculture is one of the most important sources of carbon sinks in fisheries, which has an important impact on the offshore carbon cycle. As global temperature rises and ocean acidification intensifies, the capacity of the ocean to absorb CO₂ will change. However, the effects of high temperature on the physiology and transcriptome related to carbon metabolism in Mytilus coruscus are not clear enough. This study investigated the effects of high temperatures on the total carbon content, carbon metabolism, antioxidant-related enzyme activities, and the transcriptome of Mytilus coruscus. The results showed that high temperature significantly inhibited the activities of hexokinase and pyruvate kinase, increased carbonic anhydrase activity (P＜0.05), decreased the ATP content of digestive glands (P＜0.05), and affected glycolysis and the tricarboxylic acid cycle, leading to a significant decrease in the mussel’s ability to sequester carbon. High temperature resulted in significant (P<0.05) increases in the levels of reactive oxygen species and malondialdehyde, and enhanced the activities of superoxide dismutase and catalase. Observations by transmission electron microscopy showed that high temperatures damaged the subcellular structure of the digestive gland in Mytilus coruscus, resulting in the shrinkage of the nucleolus, swelling of the endoplasmic reticulum, and a significant reduction in the mitochondrial cristae. Comparative transcriptomic analysis showed that the upregulated DEGs were mainly enriched in protein processing in the endoplasmic reticulum, antigen processing and presentation, and MAPK signaling pathway. The downregulated DEGs were mainly enriched in necroptosis, DNA replication, and the NF-kappa B signaling pathway. In antioxidant-related DEGs, the upregulated DEGs include vitamin K epoxide reductase, peroxidases, heat shock protein 105 kD, heat shock protein 70 kD, and superoxide dismutase; The downregulated DEGs mainly included NADPH oxidase, glutathione reductase, cytochrome b-245, cytochrome P450, and quinone reductase. The upregulated genes enriched in the carbon metabolism pathway included chitinase, phosphatidylinositol 4, 5-bisphosphate 3-kinase, phosphoenolpyruvate carboxykinase, galactokinase, and inositol trisphosphate 3-kinase. The downregulated genes included aldose-1-epimerase, carbonic anhydrase, galactose mutarotase, acyl-CoA synthetase, alcohol dehydrogenase, and hexokinase. In conclusion, high temperature has an inhibitory effect on the activities of enzymes and the expression of genes related to carbon metabolism in Mytilus coruscus. The results of this study are intended to provide a scientific basis for the healthy development of mussel aquaculture and the assessment of carbon sinks.

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.

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.

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.

Subunit 5B of constitutively photomorphogenic 9 signalosome (CSN5B) is an inhibitory factor for the biosynthesis of vitamin C in the L-galactose synthesis pathway in plants. To create mutants with richer vitamin C in tomato fruits, a dual-target vector of pKSE402-SlCSN5B was constructed and transformed into the breeding parent of 1912. Based on the molecular biological assay and DNA sequencing, 17 transgenic positive lines were determined, and 6 lines of them were genetically mutated at the target site of SlCSN5B, with an editing efficiency of 35.3%. Among the mutants, 2 lines were homozygous mutants, csn5b-6 with 180 bp deletion and csn5b-8 with 3 bp deletion. The biological traits of two types of deficiency homozygotes without exogenous T-DNA insertion derived from the T₁ generation were observed, and there were no significant differences in phenotypes of the plant and fruit. Through physiological testing of red-ripe fruits from T₁ generation lines of csn5b-6, the GMPase activity and the vitamin C content significantly increased by 43% and 37.8%, respectively, the content of hydrogen peroxide significantly decreased by 25.9%, and the content of soluble solids did not obviously change, compared with the wild type. There were no significant differences in plant traits and physiological characteristics between T₁ generation lines of csn5b-8 and the wild type. Based on gene expression analysis and protein structure prediction, the results showed that the gene of SlCSN5B normally transcribed, and the loss of large peptide segments of CSN5B encoded by SlCSN5B caused changes in its structure to affect functioning, which led to an increase of vitamin C content in the line of csn5b-6-11. The findings suggest that the vitamin C content of tomato fruit can be improved by CRISPR/Cas9-mediated SlCSN5B gene editing, which provides the valuable resources for high-quality breeding in tomato.