On-target, off-tumor toxicity presents a significant challenge in chimeric antigen receptor (CAR) T therapy for solid tumors. Traditional approaches to managing adverse reactions, such as suppressing in vivo CAR-T cell activity, risk impairing their antitumor efficacy, often resulting in treatment failure. Intravenously infused CAR-T cells initially traffic to the lungs, where they are activated by tumor-associated antigens (TAAs) expressed on pulmonary tissues, leading to acute lung injury. To address this, this study developed a strategy involving leukocyte function-associated antigen 1 (LFA-1) neutralization at the time of infusion. This approach modulates CAR-T cell pharmacokinetics to enhance efficacy while minimizing toxicity. Post-infusion, CAR-T cells preferentially sequester and activate in the lung, secreting tumor necrosis factor α (TNF-α), which upregulates intercellular adhesion molecule 1 (ICAM-1) expression on pulmonary endothelial cells. This triggers an "activation-adhesion" feedback loop via the LFA-1/ICAM-1 pathway, exacerbating lung injury. Neutralization of LFA-1 during infusion significantly reduces CAR-T cell adhesion to pulmonary endothelium, disrupts this feedback loop, and mitigates acute lung injury. By accelerating the pharmacokinetic progression of CAR-T cells beyond the lung, this strategy not only alleviates the acute toxicity associated with high-dose regimens but also enhances the antitumor efficacy of low-dose CAR-T cells, thus expanding the therapeutic window.

Exosomes play a crucial role in intercellular communication and disease diagnostics, yet their affinity enrichment remains challenging due to the heterogeneous expression of surface markers. To address this, we engineered a recombinant fusion protein, ExoBp, designed to bind multiple exosome surface biomarkers through integrated peptide motifs and protein domains with sterically unhindered conformation revealed by structure prediction. The constructed protein was expressed but required denaturing conditions for purification. Despite refolding challenges, ExoBp was successfully immobilized onto streptavidin-coated magnetic beads in its denatured state, followed by on-bead refolding to restore functionality. The resulting ExoBp magnetic beads demonstrated efficient exosome binding, as confirmed by the detection of exosomal markers after coincubation with exosome derived from different types of cells. Both dialysis- and dilution-based refolding methods yielded stable ExoBp-bead complexes with retained binding capacity. This study presents a novel, affinity-based exosome enrichment tool leveraging multitarget binding, offering a promising platform for exosome isolation and downstream biomedical applications.

A lateral flow analysis (LFA) strip for detecting dual miRNAs (miR155 and miR21) was constructed based on the “AND” logic gate and DNA self-assembly. The ternary synergistic strategy of “logic gating-signal amplification-visual output” enables highly sensitive and visual detection of miR21 and miR155 for breast cancer diagnosis. Only if both miR21 and miR155 are present can the trigger DNA (TDNA) in the sensor specifically dissociate and activate the “AND” logic gate (output signal = “1”); in the absence of either miRNA, the system remains in an inactive state (output signal = “0”). TDNA initiates catalytic hairpin assembly (CHA), and the product of the CHA reaction cascaded triggers the hybridization chain reaction (HCR) to achieve dual signal amplification. Gold nanoparticles (AuNPs) were used to label DNA for the construction of the LFA strip, allowing naked-eye observation of the detection results of miR21 and miR155. Detection results showed that the signal intensity of the test line (T line) had a positive correlation with the concentration of miRNAs (with a miR155:miR21 ratio of 1:1). Within the concentration range of 0.1–10 nM, the average grayscale value of the T line exhibited a good correlation with the logarithm of miRNA concentration (correlation coefficient r = 0.9983), and the limit of detection (LOD) was 30.98 pM. This LFA strip has been preliminarily validated to the detection of human serum samples, and the results indicated that the levels of miR155 and miR21 in the serum of breast cancer patients were significantly higher than those in normal individuals. Free from the limitation of large-scale instruments, this strip holds great commercialization potential.

Rationale:​​ CAR-T cell therapy has demonstrated remarkable promise for managing specific autoimmune disorders. However, it remains unclear, whether long-term immunosuppressive therapy in autoimmune patients adversely affects the phenotype and function of patient-derived CAR-T products. This study aimed to compare the characteristics of T cells and manufactured CAR-T cells from patients with multiple myeloma (MM) and chronic inflammatory demyelinating polyneuropathy (CIDP). ​​Methods:​​ T cells isolated from MM and CIDP patients, as well as healthy volunteers (for baseline comparisons only), were analyzed. CAR-T cells were generated using an identical manufacturing process. A comprehensive analysis was conducted, including flow cytometry for phenotypic and functional assessment, transcriptomic profiling via RNA sequencing, and in vitro functional assays such as cytokine secretion and cytotoxicity tests. ​​Results:​​ T cells from CIDP patients showed phenotypes and functional profiles more comparable to those from healthy volunteers. In contrast, MM-derived T cells showed increased CD8⁺ T cell frequency, elevated exhaustion markers, reduced naïve and less-differentiated subsets, and enhanced effector molecule production upon non-specific stimulation. CAR-T manufacturing reduced these inherent differences, yielding similar differentiation states, transcriptomic profiles, and convergent cytotoxic capacities. However, distinct immunomodulatory features persisted, as CIDP-derived CAR-T cells displayed reduced activation markers and lower IFN-γ secretion upon antigen stimulation compared to MM-derived CAR-T cells. ​​Conclusions:​​ Our study reveals that CAR-T manufacturing process can reduce pre-existing T-cell heterogeneity across different patient populations. These findings support the feasibility of autologous CAR-T therapies in immunosuppressed autoimmune patients, demonstrating that critical cytolytic functions are preserved despite residual alterations in cytokine profiles.

Evidence-augmented and reliable cell-type annotation remains a major bottleneck in single-cell RNA-seq analysis, particularly for rare, transitional, and disease-associated populations. To address this, we introduce scMarkerAgent, an evidence-grounded cell marker resource developed using an LLM-assisted literature-curation framework. It integrates 294,692 full-text publications to provide 890,296 high-quality cell type–marker annotations from 50,233 cell types across human, mouse, and rat. scMarkerAgent integrates 82,165 curated negative-marker annotations and 417,812 disease-context annotations, improving disambiguation of homologous cell types and delineation of malignant cells. Every cell type–marker annotation is directly supported by sentence-level literature evidence. In the cell annotation workflow, candidate labels are further refined through an LLM-based reasoning step that jointly evaluates positive and negative markers. Compared with existing resources, scMarkerAgent offers broader coverage of markers, tissues, cell types, and diseases. It is released as a FAIR-compliant database together with a code-free web platform that supports marker retrieval, automated cell annotation, and customizable cell scoring (available at https://www.markeragent.net).

Caffeic acid is a phenolic acid compound widely present in nature. While beneficial to human health within a certain concentration range, excessive intake may lead to a series of health issues, such as liver and kidney function impairment. Therefore, developing a method for the precise and rapid detection of caffeic acid concentration in serum is of great significance for guiding rational clinical medication, drug monitoring, and daily health management. This study prepared a molecularly imprinted polymer (MIP) fluorescent probe for caffeic acid detection. Blue fluorescent carbon dots (bCDs) were synthesized via a one-step hydrothermal method using citric acid and diethylenetriamine as precursors. Subsequently, a molecularly imprinted silica shell was constructed on the bCDs' surface via a sol-gel method to obtain the bCDs@MIP fluorescent probe. Leveraging the specifically tailored recognition cavities created by molecular imprinting technology, the probe selectively captures caffeic acid, leading to fluorescence quenching. The degree of fluorescence intensity reduction exhibits a positive correlation with caffeic acid concentration, enabling its quantitative detection. The limit of detection (LOD) for caffeic acid was 0.15 µM. The probe was successfully applied to the determination of caffeic acid in human serum, providing a promising fluorescent strategy for caffeic acid analysis.

Acid-sensitive GOx@MPN@Gel (GOx, luteolin, Fe, MOFs) drives glucose oxidation to fuel enhanced chemodynamic/photodynamic ROS for synergistic cancer therapy.

Background:Ischemic heart disease (IHD) among young adults represents an emerging global health concern, yet comprehensive epidemiological assessments remain limited. This study aimed to quantify the global burden of early-onset IHD and project future trends through 2046. Methods:We analyzed data from the Global Burden of Disease (GBD) 2021 study, examining IHD burden among adults aged 15-44 years across 204 countries and territories from 1990 to 2021. Age-standardized rates were calculated using WHO standard population weights. Temporal trends were assessed using estimated annual percentage change (EAPC) methods. Age-Period-Cohort models were employed to project burden through 2046. DALYs were disaggregated into Years Lived with Disability (YLDs) and Years of Life Lost (YLLs). Results:Globally, age-standardized incidence rates increased modestly from 36.54 (95% UI: 21.69-54.31) per 100,000 in 1990 to 39.18 (95% UI: 23.32-58.20) per 100,000 in 2021, representing a 7.2% increase. Despite modest rate changes, absolute incident cases increased substantially from 1.26 millions to 2.17 millions. Age-standardized mortality rates declined significantly by 15.2%, from 12.21 (95% UI: 11.57-12.88) to 10.35 (95% UI: 9.68-11.04) per 100,000. Disaggregated DALYs analysis revealed divergent trends: YLDs rates increased (EAPC: 0.39%) while YLLs rates declined (EAPC: -0.66%), reflecting improved acute survival but growing chronic disease burden. Men consistently demonstrated higher burden across all measures, with male-to-female ratios ranging from 1.7:1 for incidence to 2.7:1 for mortality in the 40-44 years age group. Substantial regional heterogeneity was observed, with East Asia showing the steepest incidence increases (EAPC: 0.63%) while High-income North America demonstrated declining trends (EAPC: -2.4%). Central Europe achieved the most substantial improvements in both mortality decline (EAPC: -2.16%) and overall disease burden reduction (EAPC: -4.46%). Projections indicate continued increases in incidence and prevalence through 2046, with incident cases reaching 2.58 millions and prevalent cases reaching 12.7 millions globally. Conclusions:Early-onset IHD represents a growing global health challenge characterized by increasing incidence and prevalence but improving survival outcomes. The substantial sex and regional disparities, coupled with projected increases in absolute burden, underscore the urgent need for balanced strategies addressing both acute care improvements and long-term disability prevention.

Chimeric antigen receptor (CAR)-T cell therapy, which utilizes genetic engineering techniques to modify T-cells to achieve specific targeting of cancer cells, has made significant breakthroughs in cancer treatment in recent years. All marketed CAR-T products are second-generation CAR-T cells containing co-stimulatory structural domains, and co-stimulatory molecules are critical for CAR-T cell activation and function. Although CD28-based co-stimulatory molecules have demonstrated potent cytotoxicity in the clinical application of CAR-T cells, they still suffer from high post-treatment relapse rates, poor efficacy durability, and accompanying severe adverse reactions. In recent years, researchers have achieved specific results in enhancing the anti-tumor function of CD28 by mutating its signaling motifs, combining the co-stimulatory structural domains, and modifying other CAR components besides co-stimulation. This paper reviewed the characteristics and roles of CD28 in CAR-T cell-mediated anti-tumor signaling and activation. We explored potential strategies to enhance CAR-T cell efficacy and reduce side effects by optimizing CD28 motifs and CAR structures, aiming to provide a theoretical basis for further clinical CAR-T cell therapy development.

BACKGROUND:The heterogeneity of Neuroblastoma (NB) leads to variation in response to treatment and outcomes. The aim of the current study is to discover AI-empowered cellular morphometric biomarkers (CMBs), to establish the corresponding CMB risk score (CMBRS), CMB risk group (CMBRG), large language model driven CMB risk score (CMB-LLM-RS), and large language model driven CMB risk group (CMB-LLM-RG), and to investigate and validate their prognostic and predictive power in NB. METHODS:In this study, the retrospective cohort enrolled 84 primary NBs between 1/2020 and 12/2021, followed up through 11/22/2024; the prospective cohort enrolled 67 primary NBs between 1/2022 and 7/2023, followed up through 11/22/2024. RESULTS:We identified 9 CMBs from a retrospective NB cohort, enabling the CMBRS, CMBRG, CMB-LLM-RS, and CMB-LLM-RG. Both CMBRG and CMB-LLM-RG are significantly associated with prognosis (p < 0.0001) and treatment response (p < 0.0001). Furthermore, we double-blindly validated the predictive power of CMBRG and CMB-LLM-RG in a prospective NB cohort, which confirms their potential value in real clinical settings. Importantly, CMBRG provides clinical value independent of the International Neuroblastoma Risk Group (INRG) classification system in both retrospective and prospective NB cohorts (p < 0.05); and the combination of CMBRG and INRG significantly increases prognostic and predictive performance for NB patients. CONCLUSIONS:These findings suggest that CMBRG and CMB-LLM-RG have prognostic and predictive value for NB and warrants evaluation in larger multicenter cohorts.

Chimeric antigen receptor (CAR)-T cell therapy has revolutionized the treatment of hematologic malignancies. However, it continues to encounter significant obstacles, including treatment relapse and limited efficacy in solid tumors. While effector T cells exhibit robust cytotoxicity, central memory T cells and stem cell-like T cells are essential for in vivo expansion, long-term survival, and persistence. Strategies such as genetic engineering to enhance CAR-T cell efficacy and durability are often accompanied by increased safety risks, which not only raise regulatory approval thresholds but also escalate CAR-T production costs. In contrast, optimizing ex vivo manufacturing conditions represents a more straightforward and practical approach, offering the potential for rapid application to commercially approved CAR-T products and enhancement of their clinical outcomes. This review examines several factors that have been shown to improve T cell memory phenotype and in vivo cytotoxic activity, including cytokines, electrolytes, signaling pathway inhibitors, metabolic modulators, and epigenetic agents. The insights provided will guide the optimization of CAR-T cell industrial production. Furthermore, considerations for selecting appropriate conditions are discussed, balancing effectiveness, cost-efficiency, safety, and regulatory compliance while addressing current challenges in the field.

Precise assessment of the cytotoxic activity of engineered immune cell therapeutics, such as chimeric antigen receptor-engineered T (CAR-T) cells, is essential for their development and quality control. However, luciferase (Luc)-based viability assays, which evaluate target cell viability by overexpressing Luc tags and measuring chemiluminescent signals, may yield biased results depending on the promoter driving Luc expression. This study demonstrates that CAR-T cells can enhance cytomegalovirus (CMV) promoter-driven transcription in target cells via the interferon-gamma (IFN-γ)/nuclear factor κB (NF-κB) signaling pathway, leading to elevated Luc expression and a discrepancy between chemiluminescent signals and actual target cell death. These findings underscore the limitations of CMV promoters in functional protein overexpression systems in the context of engineered T cell killing of target cells due to their susceptibility to transcriptional interference. Statistical analyses indicate that Luc expression driven by the elongation factor-1 alpha (EF1α) promoter exhibits the highest concordance with flow cytometry-based quantification across three CAR-T cytotoxicity assay platforms, making it a more reliable choice for evaluating CAR-T cell cytotoxicity. This study highlights the necessity of selecting appropriate promoters to ensure accurate Luc-based cytotoxicity assessments and provides critical insights for standardizing detection methodologies in CAR-T cell evaluation.

BACKGROUND:Vitamin D is a fat-soluble secosteroid that plays essential roles in calcium homeostasis, bone metabolism, and numerous other physiological processes. Vitamin D deficiency has been associated with increased risk of various diseases and mortality. However, population-based studies examining the relationship between vitamin D and mortality across different age groups remain limited. METHODS:To investigate the correlation between 25-hydroxyvitamin D [25(OH)D] levels, vitamin D status, and mortality in a cohort of 47,478 individuals aged 18-85 years. RESULTS:Higher 25(OH)D levels were associated with lower mortality risk. Compared to the vitamin D deficiency group, the Hazard ratios (HR) for all-cause mortality were 0.71 (95%CI: 0.66-0.76) in the insufficiency group and 0.64 (95%CI: 0.58-0.70) in the sufficiency group. The association varied by age: strongest in adults aged 40-59 years (HR: 0.74, 95% CI: 0.65-0.85), significant in those ≥60 years (HR: 0.86, 95% CI: 0.82-0.90), but non-significant in those aged 18-39 years. The RCS analysis revealed a non-linear relationship between 25(OH)D and mortality, with significant risk reduction observed between 59.25-261.45 nmol/L for the overall population. The optimal 25(OH)D levels (lowest HR) varied by subgroups: 96.81 nmol/L for the overall population, 102.9 nmol/L for females, 67 nmol/L for ages 40-59, and 104.23 nmol/L for ages ≥60 years, while no significant association was found in ages 18-39 years. CONCLUSION:Our findings suggest that Vitamin D are associated with mortality among the whole population. Individuals aged 40-59 may derive potential benefits from vitamin D supplementation.

Immune checkpoints are critical for maintaining autoimmune homeostasis and are implicated in various autoimmune diseases, with their significance increasingly recognized. Investigating the functions and mechanisms of these checkpoints is essential for the development of more effective treatments. Leukocyte immunoglobulin-like receptor subfamily B member 4 (LILRB4) stands out as a unique immune checkpoint, with limited expression in most normal tissues but prominent presence in various hematological and solid tumors. It is also expressed on numerous immune and stromal cells, functioning as both a “Tumor Immune Checkpoint” and a “Tumor Stromal Immune Checkpoint.” Due to its distinct expression profile, LILRB4 plays a pivotal role in tumors, autoimmune diseases, allergic reactions, and the maintenance of immune homeostasis during transplantation and pregnancy. A thorough understanding of its ligands, functions, mechanisms, and ongoing therapeutic strategies targeting LILRB4 will be crucial for the development of advanced therapeutic options. This review examines LILRB4 expression and function across multiple diseases and discusses therapeutic approaches targeting LILRB4 in various contexts. Additionally, the potential of combining current drugs with LILRB4-targeted therapies is explored. Challenges in developing LILRB4-targeting drugs are also addressed, offering valuable insights for future research.

On-target off-tumor effects precipitate severe adverse reactions in patients, significantly hindering the application of chimeric antigen receptor (CAR) T cells in both hematological and solid tumors. The underlying mechanisms remain elusive due to the absence of suitable preclinical models. To elucidate these mechanisms, a human epidermal growth factor receptor 2 (Her2) transgenic mouse model was developed to investigate CAR-T cell-induced on-target off-tumor effects. CAR-T cells initially migrated to the lungs, targeting alveolar epithelial cells and resulting in interferon-γ (IFN-γ)-dependent acute lung injury. Additionally, a regulatory mechanism involving IFN-γ-induced degradation of caspase-7 mRNA 5' untranslated regions (UTR), which amplifies acute lung injury mediated by CAR-T cells, was identified. Consequently, a strategy was validated to antagonize IFN-γ during CAR-T cell infusion, thereby mitigating acute lung injury without compromising antitumor efficacy. These findings elucidate the mechanisms of CAR-T cell-induced acute lung injury and demonstrate the viability of targeting IFN-γ to prevent this adverse reaction.

​​Triple-negative breast cancer (TNBC) patients exhibiting high PD-L1 expression demonstrate poor responses to anti-PD-L1 therapy and aggressive lung metastasis. The paradoxical role of PD-L1 beyond its immune checkpoint function and the impact of interferon-γ-secreted during immunotherapy-on metastasis remain poorly understood. Integrated reanalysis of single-cell RNA sequencing (scRNA-seq) data from TNBC lung metastases identified enriched signaling pathways. IFN-γ function was assessed using murine and human TNBC cell lines, employing in vitro assays and in vivo modeling in both immunocompetent and immunodeficient mice. CRISPR/Cas9-mediated PD-L1 ablation, pharmacological inhibitors, RNA sequencing (RNA-seq), chromatin immunoprecipitation sequencing (ChIP-seq), co-immunoprecipitation (Co-IP), bioinformatics analyses, and in vivo metastasis assays were utilized to dissect underlying mechanisms. scRNA-seq revealed significant enrichment of IFN-γ signaling within a distinct metastatic TNBC cluster. IFN-γ pretreatment potently enhanced the lung metastatic capacity of TNBC cells in both immunocompetent and immunodeficient murine models. CRISPR/Cas9-mediated PD-L1 ablation abolished IFN-γ-driven metastasis without affecting proliferation, indicating an immune checkpoint-independent mechanism. Mechanistically, IFN-γ facilitated HDAC2-mediated deacetylation of PD-L1, promoting its nuclear translocation. RNA-seq identified lymphocyte antigen 6 complex locus E (LY6E) as a key downstream effector, with expression correlating with PD-L1 in TNBC patient samples. Nuclear PD-L1 bound to the RNA polymerase II subunit POLR2A to form a transcriptional complex that directly activated LY6E expression, thereby driving metastatic dissemination. Our findings unveil a novel IFN-γ-nuclear PD-L1/POLR2A-LY6E signaling axis critical for TNBC lung metastasis. This immune-independent mechanism, driven by nuclear PD-L1 transcriptional activity, provides a mechanistic basis for the limited efficacy of anti-PD-L1 antibodies against metastasis and nominates nuclear PD-L1 complexes and LY6E as potential therapeutic targets to overcome metastatic resistance in TNBC.

The Hippo pathway is a conserved tumour suppressor signalling pathway, and its dysregulation is often associated with abnormal cell growth and tumorigenesis. We previously revealed that the transcriptional coactivator Yes-associated protein (YAP), the key effector of the Hippo pathway, is a molecular target for glioblastoma (GBM), the most common malignant brain tumour. Inhibiting YAP with small interfering RNA (siYAP) or the specific inhibitor verteporfin (VP) can diminish GBM growth to a certain degree. In this study, to enhance the anti-GBM effect of siYAP and VP, we designed stepwise-targeting and hypoxia-responsive liposomes (AMVY@NPs), which encapsulate hypoxia-responsive polymetronidazole-coated VP and DOTAP adsorbed siYAP, with angiopep-2 (A2) modification on the surface. AMVY@NPs exhibited excellent blood‒brain barrier crossing, GBM targeting, and hypoxia-responsive and efficient siYAP and VP release properties. By inhibiting the expression and function of YAP, AMVY@NPs synergistically inhibited both the growth and stemness of GBM in vitro. Moreover, AMVY@NPs strongly inhibited the growth of orthotopic U87 xenografts and improved the survival of tumour-bearing mice without adverse effects. Specific targeting of YAP with stepwise-targeting and hypoxia-responsive liposome AMVY@NPs carrying siYAP and VP efficiently inhibited GBM progression. This study provides a valuable drug delivery platform and creative insights for molecular targeted treatment of GBM in the future.

CRISPR-based screens have discovered novel functional genes involving in diverse tumor biology and elucidated the mechanisms of the cancer pathological states. Recently, with its randomness and unbiasedness, CRISPR screens have been used to discover effector genes with previously unknown roles for AML. Those novel targets are related to AML survival resembled cellular pathways mediating epigenetics, synthetic lethality, transcriptional regulation, mitochondrial and energy metabolism. Other genes that are crucial for pharmaceutical targeting and drug resistance have also been identified. With the rapid development of novel strategies, such as barcodes and multiplexed mosaic CRISPR perturbation, more potential therapeutic targets and mechanism in AML will be discovered. In this review, we present an overview of recent progresses in the development of CRISPR-based screens for the mechanism and target identification in AML and discuss the challenges and possible solutions in this rapidly growing field.

Based on the concept of "Evolutionary Traps", targeting survival essential genes obtained during tumor drug resistance can effectively eliminate resistant cells. While, it still faces limitations. In this study, lapatinib-resistant cells were used to test the concept of "Evolutionary Traps" and no suitable target stand out because of the identified genes without accessible drug. However, a membrane protein PDPN, which is low or non-expressed in normal tissues, is identified as highly expressed in lapatinib-resistant tumor cells. PDPN CAR-T cells were developed and showed high cytotoxicity against lapatinib-resistant tumor cells in vitro and in vivo, suggesting that CAR-T may be a feasible route for overcoming drug resistance of tumor based on "Evolutionary Trap". To test whether this concept is cell line or drug dependent, we analyzed 21 drug-resistant tumor cell expression profiles reveal that JAG1, GPC3, and L1CAM, which are suitable targets for CAR-T treatment, are significantly upregulated in various drug-resistant tumor cells. Our findings shed light on the feasibility of utilizing CAR-T therapy to treat drug-resistant tumors and broaden the concept of the "Evolutionary Trap".

Low temperature is one of the main abiotic stresses that affects plant growth, causing serious damage or even death to plants. The differential expression of the TaEXPA19 gene in the above and underground parts of winter wheat and the implications for cold resistance remain unclear. In this study, the TaEXPA19 gene was cloned and analysed for expression in winter wheat, and transgenic Arabidopsis thaliana was constructed to investigate the effect of the TaEXPA19 gene in response to low-temperature stress on plant growth. The TaEXPA19-A and TaEXPA19-D genes have different response patterns in the above and underground parts of transgenic A. thaliana. When plants were subjected to low-temperature stress, the leaves were quickly upregulated and the roots were downregulated, and then upregulated to respond to low-temperature stress to promote the growth of leaf length and leaf width petiole length. The results indicated that TaEXPA19 genes could improve low-temperature tolerance in plants. The results of this study laid a foundation for the study of the cold resistance of winter wheat.