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.

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

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

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

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

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

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

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

Set against the artificial intelligence (AI)-driven transformation of the educational landscape, this paper discusses how the Molecular Biology course is implementing a question-driven pedagogical reform, centering on fostering innovation as its primary goal. We introduce the educational philosophy of “Co-evolution of Inquiry and Learning”, positing that questioning and knowledge acquisition function as interdependent strands—analogous to DNA’s double helix—mutually catalyzing cognitive development and igniting innovative potential. Guided by this conceptual framework, a comprehensive instructional system has been constructed. At the content level, a three-level question bank (Foundational Cognition-Comprehensive Application-Innovative Exploration) incorporates AI tools to streamline knowledge acquisition, enhance complex problem-solving capabilities,and facilitate cutting-edge research exploration. Methodologically, the four-step teaching method (ContextualQuestioning-Autonomous Inquiry-Collaborative Discussion-Reflective Questioning) systematically activates the question bank through sequenced implementation, enabling students’transition from responding to instructor-initiated questions to generating original inquiries. The formative assessment system examines five dimensions of students’competencies while providing timely diagnostic feedback. Through two years of implementation, this AI-enhanced “inquiry-initiated, mutually reinforcing” pedagogical model has successfully transformed Molecular Biology course instruction from knowledge transmission to cognitive cultivation, demonstrating significant improvements in students’scientific reasoning, self-regulated learning capacities, and innovative practical abilities. This teaching innovation offers a forward-looking and implementable pathway for reforming core STEM curricula in the modern era.

Methcathinone, a widely abused synthetic cathinone, poses a significant public health risk because of its severe neurotoxicity and high addictive potential. In this study, we used a zebrafish model to investigate the mechanisms underlying its neurobehavioural effects. Methcathinone exposure reduced larval survival and induced a range of abnormal behaviours. Most notably, conditioned place preference (CPP) tests demonstrated the potent rewarding properties of methcathinone, as shown by a significant 22.2% increase in time spent in the drug-paired light zone compared with baseline(P<0.01). Transcriptomic analysis of brain tissue revealed systemic disruption of the neuroactive ligand-receptor interaction pathway. Gene set enrichment analysis (GSEA) further revealed significant suppression of γ-aminobutyric acid (GABA) signalling (NES =-1.64, P<0.05) and glutamate receptor signalling, including ionotropic and AMPA receptor signalling (P<0.05). Quantitative PCR validation confirmed the marked downregulation of key genes involved in these pathways: the mRNA expression of GABAergic receptors(e.g., gabra1 and gabra2), glutamatergic receptors (e.g., gria2 and grin2B), and the dopamine transporter slc6a3 decreased by 67.0% to 97.9% (all P <0.01). These results suggest that methcathinone drives reward-seeking behaviour through a synergistic dual-target mechanism; specifically, the concurrent suppression of GABAergic inhibition and slc6a3-mediated reuptake likely facilitates dopaminergic hyperactivity, whereas the downregulation of glutamate receptors reflects a homeostatic response to overstimulation. Our findings provide novel mechanistic insight into the development of methcathinone use disorder in humans.

Atherosclerosis (AS) is a vascular disease characterized by lipid deposition, chronic inflammation, and plaque formation, serving as the primary pathological basis for cardiovascular events. Although conventional lipid-lowering therapies have achieved certain clinical efficacy, the underlying core pathogenesis remains to be further elucidated. In recent years, the gut-brain axis, a bidirectional regulatory network connecting the gastrointestinal tract and central nervous system, has gradually become a research hotspot in AS due to its pivotal role in the interaction between neuroendocrine immunity and metabolism. The gut-brain axis influences AS progression through the gut microbiota-derived metabolites such as short chain fatty acids (SCFAs), trimethylamine N-oxide (TMAO) and intestinal hormones, which modulate host inflammation, lipid metabolism, and endothelial function; the autonomic nervous system (sympathetic/parasympathetic) affects plaque stability by regulating immune cell activity and inflammatory cytokine release. Dysregulated neuro-metabolic crosstalk may exacerbate microbial imbalance and metabolic disturbances, promoting a vicious cycle of AS progression. This review systematically summarizes the complex bidirectional signaling mechanisms between gut microbiota and the host nervous system, as well as their roles in the pathogenesis and progression of AS. It explores how gut microbiota regulate the host’s neuroendocrine and immune responses through metabolic products, thereby influencing the function of both the central and peripheral nervous systems; conversely, neural activities can also directly or indirectly modulate the homeostasis of the gut microbiota. Finally, the paper further outlines interventional strategies targeting the gut-brain axis for the treatment of AS and discusses their clinical feasibility. These findings provide theoretical foundations for developing precision prevention and treatment strategies centered on gut-brain axis regulation, while also identifying promising new targets and research directions for clinical interventions.

Dysregulation of cholesterol homeostasis plays a pivotal role in tumorigenesis, progression, and immune evasion. This review summarizes the core regulatory mechanisms of intracellular cholesterol metabolism, including biosynthesis mediated by sterol regulatory element-binding protein 2 (SREBP2) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), low-density lipoprotein receptor (LDLR)-dependent exogenous uptake, ATP-binding cassette transporter A1/G1 (ABCA1/ABCG1)-driven efflux, and esterification catalyzed by acyl-CoA:cholesterol acyltransferase 1/2 (ACAT1/2). It further systematically elaborates on the multidimensional regulatory functions of these metabolic pathways within the tumor immune microenvironment (TIME). In the TIME, tumor cells reprogram their own and surrounding immune cells’ cholesterol metabolism to establish an immunosuppressive milieu. Specifically, enhanced ACAT1 activity leads to cholesterol ester accumulation in T cells, impairing receptor clustering and effector function. Oxysterols, such as 25-hydroxycholesterol (25-HC) and 27-hydroxycholesterol (27-HC), modulate macrophage polarization, inhibit dendritic cell (DC) migration, and induce T cell exhaustion via liver X receptor (LXR) signaling. The functional maturation of myeloid-derived suppressor cells (MDSCs) is also regulated by the XBP1-cholesterol axis. Moreover, the antitumor activity of natural killer (NK) cells is negatively regulated by LDLR-mediated cholesterol uptake and the bile acid metabolite iso-lithocholic acid (iso-LCA). Targeting cholesterol metabolism demonstrates significant antitumor potential: statins inhibit the mevalonate pathway; inhibitors of proprotein convertase subtilisin/kexin type 9 (PCSK9) or niemann-pick C1-like 1 (NPC1L1) block cholesterol uptake; and the ACAT inhibitor avasimibe not only directly suppresses tumor growth but also reverses immunosuppressive states. Notably, combining cholesterol metabolism modulators with immune checkpoint inhibitors produces synergistic effects, significantly enhancing antitumor immunity. Therefore, cholesterol metabolism serves not only as a metabolic foundation for tumor cell proliferation but also as a critical metabolic hub regulating TIME function. Its precise intervention holds promise for developing novel and effective combinatorial strategies for cancer therapy.

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

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

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

With the accelerating pace of global aging, senescence and its associated diseases have emerged as significant public health challenges. The gut, a crucial organ in the regulation of aging, undergoes a process characterized by a loss of gut microbiota diversity, impaired intestinal barrier function, and disrupted immune regulation. These changes subsequently lead to increased oxidative stress and a systemic inflammatory state, ultimately accelerating overall organismal aging and the onset and progression of various diseases. This review systematically summarizes the mechanisms and potential applications of probiotics in alleviating gut aging. Research indicates that probiotics can mitigate the intestinal aging process via multiple pathways, including enhancing intestinal barrier function, modulating immune and antioxidant activity, regulating gut-brain axis communication, and restoring gut microbiota diversity. Furthermore, this article reviews the clinical applications of probiotics in age-related diseases, such as improving metabolic diseases (e.g., NAFLD) via the “gut-liver axis”, alleviating digestive disorders through microbiome restoration, counteracting musculoskeletal decline via the “gut-muscle/bone axis”, delaying age-related immunosenescence through immune modulation, ameliorating neurodegenerative diseases via the “gut-brain axis”, and slowing skin aging through the “gut-skin axis”. Despite the promising potential demonstrated by existing studies, challenges remain, including strain-specific effects, individual variability, and long-term safety concerns. Future research should integrate multi-omics technologies, personalized intervention strategies, and the development of novel formulations to promote the precise application of probiotics in the field of healthy aging. This review aims to provide a theoretical reference and future perspectives for mechanistic research and clinical practice concerning probiotic interventions for gut aging and related diseases.

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

Mitochondria are the energy centers of cells, and their gene expression regulation is essential for maintaining cellular homeostasis. The mitochondrial genome is a circular double-stranded DNA molecule, and its transcription depends on a transcriptional machinery composed of multiple nuclear-encoded proteins. During transcription initiation, POLRMT uses NAD as a non-canonical initiation nucleotide at the start of transcription, forming a 5′NAD cap, which links transcription with the cell′s metabolic state. The mitochondrial DNA is transcribed into long, continuous polycistronic precursor transcripts, which require subsequent processing and modification to become mature. These steps take place in mitochondrial RNA granules, where they regulate RNA stability and translation efficiency. Once matured, RNA not only stays in the mitochondrial matrix but can also be transported to the cytoplasm and nucleus. This review summarizes recent advances in mitochondrial DNA transcription and post-transcriptional processing and modification, with an emphasis on the underlying mechanisms and their sub-mitochondrial localization. It further focuses on the roles of mitochondrial RNA modifications in regulating RNA stability, translation efficiency, and gene expression, and discusses the distribution and transport mechanisms of mature mitochondrial RNA. This review aims to clarify the regulatory mechanisms of mitochondrial gene expression and to provide new insights into the mechanisms underlying mitochondrial dysfunction-related diseases and potential therapeutic targets.

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

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

Tuberculosis (TB) remains a major global public health threat, and the worsening of multidrug-resistant tuberculosis has further intensified the challenges in its prevention and control. Conventional diagnostic methods are limited by their low sensitivity and significant delays, while the introduction of artificial intelligence (AI) technology offers a breakthrough solution for TB control. In terms of diagnosis, AI technologies significantly improve the efficiency of TB screening and enable accurate identification of disease manifestations. Meanwhile, AI also plays an important role in the discovery of TB biomarkers, where it identifies high-performance novel diagnostic markers through the analysis of multi-omics data. In the field of treatment, AI models can predict drug efficacy and the risk of adverse reactions, supporting personalized therapeutic strategies. In drug development, AI accelerates the discovery of drug targets and the screening of compounds for tuberculosis, and even enables the de novo design of novel drugs. This review summarizes the latest applications of AI in the diagnosis, treatment, and drug development of tuberculosis, aiming to clarify the current role of AI in TB control and outline future research directions.

Ideological and political education within university courses is essential for fulfilling the institution’s core mission of cultivating virtue through education. As artificial intelligence technology increasingly penetrates the educational domain, the need to reform and innovate curriculum-based ideological and political instruction has become pressing. Generative Artificial Intelligence (Generative AI), a significant subset of AI, presents new avenues for overcoming teaching challenges and transforming educational methodologies through its extensive database and advanced interactive understanding and dialogue capabilities. This study investigates the application of Generative AI in the ideological and political education of the course “Clinical Immunological Testing Techniques,” aiming to improve the efficacy of moral education in teaching and to achieve a seamless integration of knowledge transfer, skill development, and value guidance. Utilizing Generative AI technology, we intelligently extracted and reconstructed ideological and political content from the curriculum, creating a human-AI collaborative teaching model structured around “precise pre-class preparation—collaborative in-class inquiry—post-class reflection and internalization”. Students from the Class of 2021 (n=219) and Class of 2022 (n=217) in the Medical Laboratory Science program were designated as the control and research groups, respectively. Teaching interventions were conducted under uniform conditions, including the same teaching teams, course content, and assessment criteria. The effectiveness was evaluated by comparing the two groups across various metrics: academic performance in professional knowledge, practical skills, learning behavior data (encompassing classroom participation and AI interaction frequency), and questionnaire responses. Results indicated that the research group significantly outperformed the control group in professional knowledge and practical skills (P<0.001). Learning behavior data revealed higher AI interaction frequency and discussion participation rates in the research group. Moreover, 89.2% of students expressed a positive attitude towards AI-enhanced ideological and political education. In conclusion, the implementation of Generative AI in “Clinical Immunological Testing Techniques” has contributed to enhancing students’ professional competence and practical abilities, enabling the precise integration of ideological elements, and facilitating the subtle infusion of value guidance, thus providing a reproducible and practical model for the reform of ideological and political education in medical courses.

Intracerebral hemorrhage (ICH) is a subtype of hemorrhagic stroke with extremely high mortality and disability rates, caused by the spontaneous rupture of non-traumatic cerebral blood vessels and the subsequent infiltration of blood into brain tissue. Due to its high incidence and poor long-term prognosis, it imposes a heavy burden on patients’ lives and the economy. Therefore, minimizing the neurological deficits after injury has become an urgent problem to be solved. In recent years, the role of extracellular vesicles (EVs) in mediating complex intercellular signaling and maintaining brain tissue homeostasis has attracted attention. Most EVs have good biocompatibility, stable membrane structure, low immunogenicity, and the ability to carry various bioactive molecules, providing favorable conditions for their successful passage through the blood-brain barrier and targeted regulation of brain cells. This article systematically reviews the biological and functional characteristics of EVs, thoroughly analyzes their dual regulatory roles in the pathophysiological process of cerebral hemorrhage, interprets the application potential of EVs as biomarkers for dynamic disease monitoring in the diagnosis of cerebral hemorrhage, and highlights the latest progress and improvement strategies of EVs as drug delivery and gene editing vehicles. This review aims to provide theoretical support for the precise clinical application of EVs in the diagnosis and treatment of cerebral hemorrhage.

Exendin-4 (Exe), a glucagon-like peptide-1 (GLP-1) receptor agonist, has a protective effect on pancreatic β cells; however, its underlying mechanism is not well understood. In this study, Exendin-4 exhibited a higher antioxidant ability and protected MIN6 cells from high glucose-induced oxidative damage by reducing reactive oxygen species (ROS) production (P < 0.01), and maintaining mitochondrial function in pancreatic β cells. The beneficial effects of Exendin-4 included increased cell viability (P < 0.05) and insulin secretion (P < 0.05), as well as improved mitochondrial membrane potential (MMP) (P < 0.01) and ATP levels (P < 0.05). Additionally, Exendin-4 inhibited lactate dehydrogenase (LDH) activity (P < 0.001) and reduced intracellular malondialdehyde (MDA) levels (P < 0.01), and boosted the activities of antioxidant enzymes such as superoxide dismutase (SOD) (P < 0.01) and catalase (CAT) (P < 0.01). Glutaredoxins (Grxs) were identified as glutathione (GSH)-dependent oxidoreductases, and the Grx/GSH system is commonly referred to as the cellular antioxidant system acting in the defense of pancreatic β cells against oxidative stress and the mitochondrial damage. Our findings revealed that Exendin-4 significantly enhanced the protein expression levels of glutaredoxin 1 (Grx1), glutaredoxin 2 (Grx2) and glutathione reductase (GR) (P < 0.05), and improved the GSH/GSSG ratio (P < 0.01) and NADPH/NADP⁺ ratio (P < 0.05). These results indicate that Exendin-4 improved the function of pancreatic cells under high glucose condition. The underlying mechanism involves increasing the expression levels of key proteins in the glutaredoxin system and inhibiting the dysfunction of the Grx/GSH system, thereby reducing mitochondrial oxidative damage and functional disorders.

Driven by the new wave of technological revolution, big data, artificial intelligence (AI), and genetic technologies are profoundly integrating into the medical field, systematically transforming healthcare models and practices. As a reconstruction of the medical education system and philosophy, the “New Medical Education” (NME) initiative responds to demands of technological advancement and educational transformation. Grounded in the fundamental mission of fostering virtue through education, the curriculum team systematically explores a digital intelligence-integrated curriculum reform pathway for Medical Genetics to overcome traditional teaching limitations, including spatiotemporal constraints, insufficiently digital-intellectualized resources, and personalized teaching challenges. This is achieved through upgrading teaching resources via a “Basic-Clinical” integrated massive open online courses (MOOC), a virtual simulation experiment platform for tiered professional training, deep integration of curriculum-based ideological and political education resources, and advanced scientific research modules; constructing a smart curriculum system by integrating knowledge-problem-competency maps with AI learning companions; and designing flexibly configurable teaching plans for diverse learner needs. Empirical results demonstrate that the reform significantly enhances teaching quality and student competency, establishing an intelligent teaching model characterized by “medical-education synergy, science-education integration, and virtual-real combination.” This provides a replicable paradigm for the digital transformation of medical education, thus contributing to cultivating high-quality innovative medical talents equipped for the intelligent era.

Protein kinase C substrate 80K-H, also known as the Glucosidase Ⅱ beta subunit, interacts with the Glucosidase Ⅱ alpha subunit to jointly participate in regulating the processing and maturation of glycoproteins in the endoplasmic reticulum, and plays an important role in N-glycosylation, a key post-translational modification. At present, the close relationship between PRKCSH gene mutations and Autosomal Dominant Polycystic Liver Disease has been widely studied and concerned. The multiple functions of PRKCSH, including regulating signaling pathways such as IGF1R and IRE1α, make it a promising therapeutic target and biomarker for cancer immunotherapy. Meanwhile, in cancer, PRKCSH can affect cell growth, metastasis, and responses to growth factors through pathways such as responding to endoplasmic reticulum stress, coordinating cell death programs, and participating in immunotherapy, thereby influencing autophagy or apoptosis. PRKCSH can also enhance cancer treatment and regulate anti-tumor immunity by boosting the activity of NK cells and T cells. In addition, PRKCSH can generate different isoforms through alternative splicing, affecting processes such as epithelial-mesenchymal transition and lung cancer cell proliferation. Although PRKCSH has been identified in various diseases, more evidence is still needed to clarify its specific mechanism of action and development potential.

Protein arginine methyltransferase 1 (PRMT1), a key enzyme regulating protein arginine methylation, is involved in the development of various tumors. This study aimed to investigate the role of PRMT1 in hepatocellular carcinoma (HCC) progression and its clinical significance. We first analyzed the pan-cancer expression patterns of PRMT1 and its association with prognosis by integrating public databases such as TCGA, GTEx, and GEPIA2. Subsequently, real-time quantitative PCR (qPCR) and Western blotting were used to detect PRMT1 expression in the HCC cell line Huh7, and its subcellular localization was determined by immunofluorescence. Transwell and wound healing assays were further employed to assess the effect of PRMT1 knockdown on the migration and invasion abilities of Huh7 cells. Integrated analysis of public databases including TCGA revealed that PRMT1 expression was significantly upregulated in multiple tumor tissues (P < 0.05), and its high expression was associated with poor patient prognosis. Functional enrichment analysis indicated that PRMT1 is mainly involved in pathways such as the cell cycle and DNA repair. In cellular experiments, qPCR and Western blotting results showed that PRMT1 was highly expressed in the HCC cell line Huh7. Immunofluorescence further confirmed that PRMT1 was primarily localized in the nucleus. Transwell and wound healing assays demonstrated that knockdown of PRMT1 significantly reduced the migration and invasion abilities of Huh7 cells. These findings indicate that PRMT1 exhibits high expression and nuclear localization in HCC and regulates the migration and invasion processes of HCC cells, providing new experimental evidence for understanding the role of PRMT1 in the development and progression of HCC.

Polycystic ovary syndrome (PCOS) is one of the most prevalent endocrine, metabolic, and reproductive disorders among women of reproductive age, with increasing incidence worldwide, posing significant threats to female reproductive health and long-term quality of life. The syndrome is clinically characterized by hyperandrogenemia (HA), ovulatory dysfunction, and polycystic ovarian morphology, and is frequently accompanied by insulin resistance (IR), dyslipidemia, and a chronic low grade inflammatory state. In recent years, with advances in molecular research, ferroptosis, a novel form of iron-dependent regulated cell death, has been implicated as a key player in the pathogenesis of PCOS. Multiple types of non-coding RNA (ncRNA), including microRNA (miRNA), long noncoding RNA (lncRNA), and circular RNA (circRNA), have been shown to finely regulate ferroptosis through various mechanisms targeting iron metabolism, redox homeostasis, lipid metabolism, and related signaling pathways. This ncRNA-mediated regulatory network of ferroptosis is extensively involved in core pathological processes of PCOS, such as granulosa cell (GC) dysfunction, follicular development arrest, excessive androgen synthesis, and dysregulated insulin secretion, thereby contributing to disease progression at multiple levels. This review systematically summarizes the core molecular mechanisms of ferroptosis, highlights the specific roles of ncRNAs in modulating ferroptosis, and elaborates on the potential implications of this regulatory axis in PCOS and its related complications. Furthermore, current limitations and challenges in studying ncRNA-regulated ferroptosis in PCOS are discussed, along with prospective directions for future research. This review aims to provide new theoretical insights into the molecular pathology of PCOS and to offer potential strategies for ncRNA-based clinical diagnosis and therapeutic interventions.

Endometrial carcinoma (EC) is one of the most common malignancies of the female reproductive system, and its incidence continues to rise. While diagnostic and therapeutic approaches have advanced, the prognosis for patients with advanced-stage remains poor. Emerging treatment strategies, particularly targeted therapies, offer new hope and underscore the urgent need for novel therapeutic targets. The ubiquitin-proteasome system (UPS), a key pathway for intracellular protein degradation, significantly influences cancer progression by regulating key proteins in oncogenic pathways through ubiquitination or deubiquitination. microRNAs (miRNAs) are a class of small RNAs widely present in eukaryotic cells that regulate gene expression and the cell cycle by binding to specific mRNAs and inhibiting their posttranscriptional expression. They can also directly target key ubiquitin ligases or deubiquitinating enzymes (DUBs), affecting downstream pathways. This review discusses how ubiquitin ligases or deubiquitinating enzymes regulate related proteins in several signaling pathways in EC and explores the mechanisms by which abnormally expressed microRNAs influence oncogenesis by targeting mRNAs of proteins in the UPS. By elucidating the roles of dysregulated ubiquitination and deubiquitination from RNA and protein perspectives, this study identifies potential therapeutic strategies and provides new insights into molecular diagnostic markers and treatments for EC.

Plasmids are fundamental tools in the life sciences and biomedical research; however, developing efficient and low-cost extraction methods remains a major challenge. Here, we evaluated silica particles of diverse morphologies and sizes as DNA adsorbents, optimized the alkaline lysis buffer, and systematically refined critical parameters, including chaotropic salt concentration, silica dosage, and adsorption and elution kinetics. These optimizations substantially improved extraction performance. The resulting protocol is streamlined, inexpensive, and consistently yields high-quality plasmid DNA, with outputs comparable to those obtained using commercial spin-column kits. Moreover, the method is linearly scalable, achieving yields and purity similar to those of silica membrane-based Maxiprep columns, and is fully compatible with downstream applications such as restriction digestion and cell transfection. Collectively, this work provides a robust and scalable plasmid extraction strategy with strong potential for commercial translation.

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

Amyloid precursor protein (APP) metabolic imbalance is one of the core pathological mechanisms of Alzheimer’s disease (AD). The normal metabolism of APP depends on the dynamic balance between the non-amyloid pathway (mediated by α/γ-secretase to produce soluble APPα and p3 fragments) and the amyloid pathway (mediated by β/γ-secretase to produce Aβ peptides). In the pathological state of AD, a variety of factors such as abnormalities in genetics (such as APP, PSEN gene mutation, APOEε4 allele), epigenetics and transporters (such as LRP1, SNXs family) work together to break this balance, resulting in excessive activation of the amyloid pathway and increased production of toxic Aβ42/43 oligomers. These Aβ oligomers, as key toxic substances, eventually lead to synaptic loss and neuronal death by destroying neuronal membrane structure, interfering with synaptic receptor function and neurotransmitter balance, inducing mitochondrial dysfunction and oxidative stress, activating microglia/astrocytes-mediated neuroinflammation, promoting abnormal phosphorylation of Tau protein, and damaging cerebrovascular function and Aβ clearance ability. APP metabolic imbalance is the initial and continuous driving link of AD pathology. Although immunotherapy targeting Aβ has made progress, future research needs to focus on analyzing the fine regulation mechanism of APP metabolic network, developing etiological intervention strategies such as specific regulation of secretase activity, enhancement of Aβ clearance and use of AICD negative feedback, and integrating early lifestyle intervention to achieve effective prevention and treatment of AD. This article systematically reviews the regulatory networks of the two types of APP metabolic pathways, analyzes the key factors of their imbalance, elucidates the Aβ-mediated AD pathological cascade, and summarizes the progress and limitations of Aβ-targeted therapies.

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

Fiber quality is an important economic trait of cotton and one of the major breeding targets. In this study, 408 upland cotton accessions were genotyped using the CottonSNP80K chip, yielding 12 874 high-quality SNP linkage disequilibrium block (SNPLDB) markers. A total of 128 SNPLDBs significantly associated with fiber length, 163 with fiber strength, 149 with fiber fineness, 94 with fiber uniformity, and 92 with fiber elongation were detected across six environments (five single environments and one multi-environment) using restricted two-stage multi-locus genome-wide association analysis (RTM-GWAS). Among these, 10, 13, 9, 4 and 4 SNPLDBs associated with the five traits, respectively, were detected in at least two environments. Further single-marker/haplotype analysis revealed significant phenotypic differences among alleles of 6, 12, 9, 3 and 3 SNPLDBs associated with these five traits, respectively. A total of 11 loci associated with fiber length, 15 with fiber strength, 8 with fiber fineness, 6 with fiber uniformity, and 4 with fiber elongation identified in this study were found to overlap with QTLs/markers reported in previous studies. Among them, the loci Block_A13_74977977_75077975 (associated with fiber strength), Block_A05_24309961_24322003 and Block_D03_2459185_2528302 (associated with fiber fineness), and Block_D04_47711028_47718399 (associated with fiber elongation) were detected in multiple environments in this study as well as in previous studies. Combining GO enrichment and RNA-seq analysis indicated that 8 genes related to fiber length, 5 genes related to fiber strength, 8 genes related to fiber fineness, 4 genes related to fiber uniformity, and 3 genes related to fiber elongation may be potential candidate genes for fiber quality. This study provides a new perspective for elucidating the genetic basis of fiber quality in upland cotton and lays an important foundation for molecular breeding of fiber quality.

The structure and function of proteins are closely related. The structure of a protein determines its function, while functional demands drive the evolution and adaptation of its structure. This dynamic relationship enables proteins to meet diverse biological needs and perform a variety of key biological functions within cells. Structural motifs and domains, as fundamental units of protein structure and function, play a crucial role in the evolution of protein functions. In biochemistry education, the chapter on "protein structure and function" is a core topic, with structural motifs and domains being key concepts. However, these topics are often characterized by their theoretical abstraction and logical complexity, posing significant challenges for both teaching and learning. This paper takes the classic helix-turn-helix (HTH) motif as an example and explores the evolutionary process of structural optimization and functional diversification from a single motif to the homeodomain (HD), then to the interactions between homeodomains and to the functional versatility of the BEN domain. By analyzing this evolutionary process, the paper reveals the important role of structural motifs and domains in the evolution of protein functions and proposes specific design and implementation plans. These plans are intended to provide valuable references for the reform of biochemistry education.

The 14-3-3 family is a group of evolutionarily highly conserved proteins in eukaryotes, which typically function as dimers to interact with phosphorylated target proteins to regulate critical biological processes such as cell physiology, metabolism, and immunity. In mammals, this family comprises seven isoforms (β, ε, η, σ, γ, ζ and θ), each exhibiting distinct structural features, tissue distribution, and ligand preference, which contribute to their functional diversity. 14-3-3 binds to target proteins by recognizing phosphorylated motifs, and regulates the functions of target proteins. In cellular autophagy, 14-3-3 can modulate the autophagic process by controlling the synthesis or degradation of key autophagy-related molecules such as Beclin-1 and ATG13. In cellular apoptosis, 14-3-3 exerts anti-apoptotic effects by inhibiting the Ask1-JNK/p38 signaling pathway and preventing the dimerization of procaspase-2. In the context of disease development, 14-3-3 proteins demonstrate multifaceted biological roles. In viral infections, they exhibit dual functions—either hijacked by viruses to facilitate replication and spread or employed by the host to inactivate viral proteins and activate immune defenses. In neurodegenerative diseases, the γ and θ isoforms of 14-3-3 exert neuroprotective effects by inhibiting the kinase activity of LRRK2 and the aggregation of α-syn. In various cancers, 14-3-3 promotes tumorigenesis and development by activating pro-proliferative pathways such as MAPK/c-Jun and PI3K/Akt pathways or inhibiting apoptosis. Although 14-3-3 holds potential as a therapeutic target, and research has advanced from structural analysis to exploration of functional mechanisms, there remain issues such as the lack of systematic studies on isoform functions and unclear in vivo dynamic regulatory mechanisms. This review summarizes recent research findings on the role of 14-3-3 in regulating autophagy, apoptosis, and the development of diseases, and presents examples of emerging biological technologies applied in 14-3-3 research, providing a theoretical basis for the subsequent development of drugs targeting 14-3-3.

In-fusion cloning technology, as a revolutionary and efficient molecular biology tool, has been applied in multiple research fields such as basic biology, biotechnology, and biomedicine. In this article, we introduce a teaching reform project suitable for undergraduate students in the course of “Molecular Biology Laboratory”, which utilizes in-Fusion cloning technology to construct a prokaryotic expression vector for alkaline phosphatase mutant genes. Through specific teaching cases, we systematically explored the design and implementation of experimental projects, and focused on analyzing the key and difficult points of the teaching content. Our teaching practice has found that the implementation of this educational reform project has achieved very good results in enhancing students’ core biological literacy, bioinformatics skills, research thinking, and innovation abilities. At the same time, the application of this technology can significantly improve the quality of experimental teaching, providing new ideas and practical references for promoting the reform and innovation of National First-Class Courses.