PEGylation Can Effectively Strike a Balance in siRNA Delivery Performances of Guanidinylated Linear Synthetic Polypeptides with Potential Use for Transcriptional Gene Silencing.

The prevailing design philosophy for polymeric vectors delivering siRNA is rooted in the post-transcriptional gene silencing (PTGS) mechanism. Yet, the transcriptional gene silencing (TGS) mechanism offers a potentially more durable silencing effect, which necessitates efficient siRNA delivery into the nucleus. However, it remains a challenge for the polymeric vectors to efficiently deliver siRNA into the nucleus. We have explored guanidinylated cyclic synthetic polypeptides (GCSPs) to enhance the nuclear delivery of siRNA, but an increased cytotoxicity and difficulty in producing the GCSPs on a large scale limit their utility. Herein, we simply prepare PEGylated guanidinylated linear synthetic polypeptides (PGLSPs) exhibiting improved membrane penetration, direct siRNA transport to the nucleus, reduced toxicity, high cellular uptake, and mitigation of protein corona formation. The PEGylation can effectively balance the vector's nuclear delivery capacity with other critical aspects of performances for siRNA delivery. Therefore, the PGLSPs hold promise as TGS-based delivery vectors, offering potential for future therapeutic applications.

Transfer dynamics of antimicrobial resistance among gram-negative bacteria.

Antimicrobial resistance (AMR) in gram-negative bacteria (GNBs) is a significant global health concern, exacerbated by mobile genetic elements (MGEs). This review examines the transfer of antibiotic resistance genes (ARGs) within and between different species of GNB facilitated by MGEs, focusing on the roles of plasmids and phages. The impact of non-antibiotic chemicals, environmental factors affecting ARG transfer frequency, and underlying molecular mechanisms of bacterial resistance evolution are also discussed. Additionally, the study critically assesses the impact of fitness costs and compensatory evolution driven by MGEs in host organisms, shedding light on the transfer frequency of ARGs and host evolution within ecosystems. Overall, this comprehensive review highlights the factors and mechanisms influencing ARG movement among diverse GNB species and underscores the importance of implementing holistic One-Health strategies to effectively address the escalating public health challenges associated with AMR.

Cathepsin C exacerbates EAE by promoting the expansion of Tfh cells and the formation of TLSs in the CNS.

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) mediated by CD4+ T helper (Th) cells, and characterized by immune cell infiltration, demyelination and neurodegeneration, with no definitive cure available. Thus, it is pivotal and imperative to acquire more profound comprehension of the underlying mechanisms implicated in MS. Dysregulated immune responses are widely believed to play a primary role in the pathogenesis of MS. Recently, a plethora of studies have demonstrated the involvement of T follicular helper (Tfh) cells and tertiary lymphoid-like structures (TLSs) in the pathogenesis and progression of MS. Cathepsin C (CatC) is a cysteine exopeptidase which is crucial for the activation of immune-cell-associated serine proteinases in many inflammatory diseases in peripheral system, such as rheumatoid arthritis and septicemia. We have previously demonstrated that CatC is involved in neuroinflammation and exacerbates demyelination in both cuprizone-induced and experimental autoimmune encephalomyelitis (EAE) mouse models. However, the underlying immunopathological mechanism remains elusive. In the present study, we established a recombinant myelin oligodendrocyte glycoprotein 35-55 peptide-induced EAE model using conditional CatC overexpression mice to investigate the effects of CatC on the alteration of CD4+ Th subsets, including Th1, Th2, Th17, Tfh and T regulatory cells. Our findings demonstrated that CatC particularly enhanced the population of Tfh cell in the brain, resulting in the earlier onset and more severe chronic syndrome of EAE. Furthermore, CatC promoted the formation of TLSs in the brain, leading to persistent neuroinflammation and exacerbating the severity of EAE in the chronic phase. Conversely, treatment with AZD7986, a specific inhibitor of CatC, effectively attenuated the syndrome of EAE and its effects caused by CatC both in vivo and in vitro. These findings provide a novel insight into the critical role of CatC in innate and adaptive immunity in EAE, and specific inhibitor of CatC, AZD7986, may contribute to potential therapeutic strategies for MS.

Pectin from steam explosion-treated citrus peel exhibits good emulsion properties and bioavailability-promoting effect in vitro of nobiletin.

Steam explosion (SE) is a potential method to modify pectin structure, which might be connected to its emulsifying characteristics and the bioavailability of encapsulated polymethoxyflavone like nobiletin. However, the relationship between SE-modified pectin and the bioavailability of encapsulated nobiletin is still unclear. In this study, nobiletin-loaded emulsion was fabricated using citrus pectin modified with SE (0.15-0.9 MPa, 3 min) as emulsifier for in vitro digestion study, and the transport and absorption of nobiletin in Caco-2 cells to investigate the bioavailability-promoting effect. The results showed that SE treatment lowered the droplet size of emulsion from 21.38 ± 2.30 µm to 2.14 ± 0.12 µm, enhanced the nobiletin encapsulation efficiency from 23.73 ± 0.78% to 86.27 ± 3.81%, improved the nobiletin bioaccessibility in vitro from 2.48 ± 0.10% to 25.42 ± 0.10% and increased the intracellular accumulation of nobiletin by over 10 times, even higher than that of Tween 80. In conclusion, pectin from SE-treated citrus peel exhibited good emulsion properties and bioavailability-promoting effect in vitro of nobiletin.

Identification of critical genes associated with oxidative stress pathways in benzene-induced hematotoxicity.

Background and aims: Bone marrow failure (BMF) is chronic benzene-induced hematotoxicity, which is associated with differential gene expression abnormality. Benzene-induced BMF is characterized by irreversible bone marrow depression. Despite extensive studies have been conducted, there is a lack of reliable, useful and simple diagnostic method for BMF. Previous studies have shown that the aberrant gene expression changes and reactive oxygen species production in bone marrow cells related to the development of BMF. Early detection of differentially expressed genes (DEGs) as potential biomarkers is important for diagnosis and treatment. However, the validation of effective biomarker through DEGs analysis in benzene-induced BMF still deserve to be clarified. This study aimed to identify target genes as potential biomarkers with benzene-induced BMF based on DEGs analysis. Methods: First, we developed a benzene-induced BMF mouse model and obtained the DEGs in bone marrow cells of benzene-exposed CD1 mice. Next, after obtaining the DEGs via RNA-Sequencing (RNA-seq) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were also used, key genes associated with benzene-induced BMF were identified. Additionally, the key markers for benzene poisoning was evaluated using qRT-PCR technique. Results: We identified DEGs for further KEGG functional analysis. Ten statistically significantly (up or down) regulated genes, namely Mapk11, Foxo1, Lefty1, Ren1, Bank1, Fgf3, Cdc42ep2, Rasgrf1, P2rx7, and Shank3 were found mainly associated with mitogen-activated protein kinases (MAPK) oxidative stress pathway . Further analysis using qRT-PCR identified that eight statistically significant DEGs associated with signaling pathways such as MAPK. We found that the level of mRNA expression of Mapk11, Foxo1, Bank1, Lefty1, Ren1, P2rx7, and Fgf3 genes were increased and Cdc42ep2 gene was decreased in BMF mice compared to control mice. Additionally, we validated the eight candidate genes for potential biomarkers in peripheral blood mononuclear cells of benzene poisoning patients by qRT-PCR. Conclusion: Our results indicated that Mapk11 and Fgf3 were predominantly candidate genes linked to novel biomarkers for benzene hematotoxicity in human beings. Our study will provide new candidate genes as useful biomarkers involved in benzene-induced hematotoxicity.

Revealing the Freezing-Induced Alteration in Microplastic Behavior and Its Implication for the Microplastics Released from Seasonal Ice.

Ice can serve as a significant temporary repository and conveyance mechanism for microplastics (MPs). MPs present in the water column can become entrapped within developing ice formations, subsequently being sequestered and transported by ice floes. With changing temperatures, MPs stored in ice can be released back into the environment, while freezing conditions can alter the properties of MPs, ultimately affecting the fate of MPs in the environment. Freezing of MPs in freshwater ice results in the aggregation of MP particles due to physical compression, leading to an increase in particle size once the MPs are released from the ice. The freezing-induced aggregation enhances buoyancy effects, accelerating the settling/rising velocity of MPs in water. Additionally, freezing can lead to enhanced surface wetting alterations, thus improving the dispersion of hydrophobic MPs. The presence of salt in the water can mitigate the effect of freezing on MPs due to the formation of a brine network within the ice structure, which reduces the pressure on MPs entrapped by ice. In cold regions, numerous MPs undergo freezing and thawing, re-entering the water column.

Confirmation of a measurement model for hospital supply chain resilience.

Background: The hospital supply chain has revealed increasing vulnerabilities and disruptions in the wake of the COVID-19 pandemic, threatening the healthcare services and patient safety. The resilience of hospital supply chains has emerged as a paramount concern within the healthcare system. However, there is a lack of systematic research to develop an instrument tailored to the healthcare industry that is both valid and reliable for measuring hospital supply chain resilience. Therefore, this study aims to construct and validate a comprehensive scale for assessing hospital supply chain resilience, based on dynamic capability theory. Methods: This study followed rigorous scale development steps, starting with a literature review and 15 semi-structured interviews to generate initial items. These items were then refined through expert panel feedback and three rounds of Delphi studies. Using data from 387 hospitals in Province S, mainland China, the scale underwent rigorous testing and validation using structural equation modeling. To ensure the most effective model, five alternative models were examined to determine the most suitable parsimonious model. Results: The study produced a 26-item scale that captures five dimensions of resilience in line with dynamic capability theory: anticipation, adaptation, response, recovery, and learning, all showing satisfactory consistency, reliability and validity. Conclusion: The multi-dimensional scale offers hospital managers a valuable tool to identify areas needing attention and improvement, benchmark resilience against their counterparts, and ultimately strengthen their supply chains against unexpected risks.

Heart size variation in anurans from China does not follow the Hesse's rule.

We failed to dissect the associations between relative heart size and geographical gradients (altitude and latitude) and climatic factors (annual temperature, rainfall, and wind speed) across 116 species, which suggests that heart size variation in anurans did not follow the Hesse's rule.

Shape Memory Polyurethane Composite With Fast Response to Near-Infrared Light Based on Tannic Acid-Iron and Dynamic Phenol-Carbamate Network.

Intelligent materials derived from green and renewable bio-based materials garner widespread attention recently. Herein, shape memory polyurethane composite (PUTA/Fe) with fast response to near-infrared (NIR) light is successfully prepared by introducing Fe3+ into the tannic acid-based polyurethane (PUTA) matrix through coordination between Fe3+ and tannic acid. The results show that the excellent NIR light response ability is due to the even distribution of Fe3+ filler with good photo-thermal conversion ability. With the increase of Fe3+ content, the NIR light response shape recovery rate of PUTA/Fe composite films is significantly improved, and the shape recovery time is reduced from over 60 s to 40 s. In addition, the mechanical properties of PUTA/Fe composite film are also improved. Importantly, owing to the dynamic phenol-carbamate network within the polymer matrix, the PUTA/Fe composite film can reshape its permanent shape through topological rearrangement and show its good NIR light response shape memory performance. Therefore, PUTA/Fe composites with high content of bio-based material (TA content of 15.1-19.4%) demonstrate the shape memory characteristics of fast response to NIR light; so, it will have great potential in the application of new intelligent materials including efficient and environmentally friendly smart photothermal responder.

High burden of human papillomavirus infection among men in Guangzhou, South China: Implications for HPV vaccination strategies.

The epidemiological and clinical aspects of Human Papillomavirus (HPV) infection in women have been extensively studied. However, there is a lack of information regarding HPV characteristics in males. In this study, we conducted a retrospective and observational study of 3737 consecutive male individuals attending outpatient clinics of Guangdong Women and Children Hospital from 2012 to 2023 in Guangzhou, South China, to determine the age- and genotype-specific prevalence of HPV in men. The results showed the overall prevalence of HPV among men was 42.15% (1575/3737), with variations ranging from 29.55% to 81.31% across distinct diagnostic populations. Low-risk HPV6 (15.47%), HPV11 (8.94%), and high-risk HPV52 (5.51%) were the most common types. The annual HPV prevalence decreased significantly (Z = -3.882, p < .001), ranging from 31.44% to 52.90%. 28.77% (1075/3737) of men manifested infection with a singular HPV type, predominantly identified as a low-risk type. The age-specific distribution of HPV infections revealed distinctive peaks in the < 25 y age group (47.60%, 208/437) and the 40-44 y age group (44.51%, 154/346). Notably, the positive rate of Chlamydia trachomatis was significantly higher among HPV-positive individuals in comparison to HPV-negatives (16.14% vs. 11.25%, p < .05). Our findings reveal a substantial prevalence of HPV infection among outpatient men in Guangzhou, South China. It is recommended to consider the inclusion of HPV vaccination for adolescent males in national immunization schedules, once an adequate supply of vaccines is accessible.

Spotlight on the vertical migration of aged microplastics in coastal waters.

Coastal waters are complex and dynamic areas with unique environmental attributes that complicate the vertical migration of microplastics (MPs). The MPs that enter coastal waters from diverse sources, including terrestrial, riverine, oceanic, and shoreline inputs undergo various aging pathways. In this study, the variations in the physiochemical characteristics of MPs undergoing various aging pathways and their vertical migration under dynamic conditions subjected to the effects of different MP characteristics and coastal environmental features were comprehensively explored. Opposite effects of aging on the vertical migration of hydrophobic and hydrophilic MPs were observed, with aging appearing to promote the dispersion of hydrophobic MPs but enhance the vertical migration of hydrophilic ones. The positive role of salinity and the negative role of humic acid (HA) concentrations on MP vertical migration were identified, and the mechanisms driving these effects were analyzed. Notably, intense turbulence not only promoted the floating of positively buoyant MPs but also reversed the migration direction of negatively buoyant MPs from downward to upward. Aging-induced changes in MP characteristics had a limited effect on MP vertical migration. The inherent characteristics of MPs and the surrounding environmental features, however, played major roles in their vertical migration dynamics. ENVIRONMENTAL IMPLICATION: Microplastics (MPs) have emerged as a significant global environmental concern and the coastal zones are the hotspots for MP pollution due to their high population density. This study comprehensively investigated the variations in the physiochemical characteristics of MPs undergoing various aging pathways. Their vertical migration patterns under dynamic conditions subjected to the effects of different MP characteristics and coastal environmental features were revealed. The roles of turbulence and MP density in their migration were identified. The findings of this study have important implications for understanding the transport and determining the ecological risks of MPs in coastal waters.

Advances in the Application of Single-Cell Transcriptomics in Plant Systems and Synthetic Biology.

Plants are complex systems hierarchically organized and composed of various cell types. To understand the molecular underpinnings of complex plant systems, single-cell RNA sequencing (scRNA-seq) has emerged as a powerful tool for revealing high resolution of gene expression patterns at the cellular level and investigating the cell-type heterogeneity. Furthermore, scRNA-seq analysis of plant biosystems has great potential for generating new knowledge to inform plant biosystems design and synthetic biology, which aims to modify plants genetically/epigenetically through genome editing, engineering, or re-writing based on rational design for increasing crop yield and quality, promoting the bioeconomy and enhancing environmental sustainability. In particular, data from scRNA-seq studies can be utilized to facilitate the development of high-precision Build-Design-Test-Learn capabilities for maximizing the targeted performance of engineered plant biosystems while minimizing unintended side effects. To date, scRNA-seq has been demonstrated in a limited number of plant species, including model plants (e.g., Arabidopsis thaliana), agricultural crops (e.g., Oryza sativa), and bioenergy crops (e.g., Populus spp.). It is expected that future technical advancements will reduce the cost of scRNA-seq and consequently accelerate the application of this emerging technology in plants. In this review, we summarize current technical advancements in plant scRNA-seq, including sample preparation, sequencing, and data analysis, to provide guidance on how to choose the appropriate scRNA-seq methods for different types of plant samples. We then highlight various applications of scRNA-seq in both plant systems biology and plant synthetic biology research. Finally, we discuss the challenges and opportunities for the application of scRNA-seq in plants.

Emergence and circulation of enterovirus B species in infants in southern China: A multicenter retrospective analysis.

BACKGROUND: Enteroviruses (EV) are common and can cause severe diseases, particularly in young children. However, the information of EV infection in infants in China is limited due to the vast population size and extensive geographical area of the country. Here, we conducted a retrospective multicenter analysis of available EV data to assess the current epidemiological situation in the infant population in southern China. METHODS: The study enrolled infants with suspected EV infection from 34 hospitals across 12 cities in southern China between 2019 to 2022, and the confirmation of EV was done using RT-PCR and VP1 gene sequencing. RESULTS: Out of 1221 infants enrolled, 330 (27.03%) were confirmed as EV-infected. Of these, 260 (78.79%) were newborns aged 0-28 days. The EV belonged to three species: EV-B (80.61%), EV-A (11.82%), and human rhinovirus (7.58%). Newborns were more susceptible to EV-B than older infants (p < 0.001). Within EV-B, we identified 15 types, with coxsackievirus (CV) B3 (20.91%), echovirus (E) 11 (19.70%), and E18 (16.97%) being the most common. The predominant EV types changed across different years. EV infection in infants followed a seasonal pattern, with a higher incidence from May to August. Furthermore, perinatal mother-to-child EV transmission in 12 mother-newborn pairs were observed. CONCLUSION: Our study is the first to demonstrate the emergence and widespread circulation of EV-B species, mainly CVB3, E11, and E18, in southern China, primarily affecting young infants. This research provides valuable insights for future epidemic assessment, prediction, as well as the elimination of mother-to-child transmission.

Unveiling the Vertical Migration of Microplastics with Suspended Particulate Matter in the Estuarine Environment: Roles of Salinity, Particle Properties, and Hydrodynamics.

The estuary is an energetic area connecting the inland, river, and ocean. The migration of microplastics (MPs) in this highly complex area is tied to the entire ecosystem. In this study, the effects of cohesive SPM (clay) and noncohesive SPM (sand) on the vertical migration of positively buoyant MPs, polyethylene (PE), and negatively buoyant MPs, polytetrafluoroethylene (PTFE), in the estuarine environment under hydrodynamic disturbances were investigated. The settling of positively buoyant MPs was more reliant on the cohesive SPM compared to the settling of negatively buoyant MPs. Moreover, MPs interacting with the SPM mixture at a clay-to-sand ratio of 1:9 settled more efficiently than those interacting with clay alone. A significant positive correlation was observed between MP settling percentage and the salinity level. MP settling percentage was significantly negatively correlated with fluid shear stress for both types of MPs, meanwhile, negatively buoyant MPs were able to resist greater hydraulic disturbances. In the low-energy mixing state, for both types of MPs, the settling percentage reached about 50% in only 10 min. The resuspension process of MPs under hydrodynamic disturbances was also uncovered. Additionally, the migration and potential sites of MPs were described in the context of prevalent environmental phenomena in estuaries.

Inhibiting ferroptosis in brain microvascular endothelial cells: A potential strategy to mitigate polystyrene nanoplastics‒induced blood‒brain barrier dysfunction.

Polystyrene nanoplastics (PS-NPs), a group of ubiquitous pollutants, may injure the central nervous system through the blood‒brain barrier (BBB). However, whether exposure to PS-NPs contributes to BBB disruption and the underlying mechanisms are still unclear. In vivo, we found that PS-NPs (25 mg/kg BW) could significantly increase BBB permeability in mice and downregulate the distribution of the tight junction-associated protein zona occludens 1 (ZO-1) in brain microvascular endothelial cells (BMECs). Using an in vitro BBB model, exposure to PS-NPs significantly reduced the transendothelial electrical resistance and altered ZO-1 expression and distribution in a dose-dependent manner. RNA-seq analysis and functional investigations were used to investigate the molecular pathways involved in the response to PS-NPs. The results revealed that the ferroptosis and glutathione metabolism signaling pathways were related to the disruption of the BBB model caused by the PS-NPs. PS-NPs treatment promoted ferroptosis in bEnd.3 cells by inducing disordered glutathione metabolism in addition to Fe2+ and lipid peroxide accumulation, while suppressing ferroptosis with ferrostatin-1 (Fer-1) suppressed ferroptosis-related changes in bEnd.3 cells subjected to PS-NPs. Importantly, Fer-1 alleviated the decrease in ZO-1 expression in bEnd.3 cells and the exacerbation of BBB damage induced by PS-NPs. Collectively, our findings suggest that inhibiting ferroptosis in BMECs may serve as a potential therapeutic target against BBB disruption induced by PS-NPs exposure.

Prolonged drying impedes the detachment of microplastics in unsaturated substrate: Role of flow regimes.

The detachment of microplastics (MPs) from porous media under different moisture conditions and flow regimes has garnered limited attention within the research community. The present study investigates the detachment of MPs from porous media under wet and dry conditions combined with steady and transient flow. For both the wet and dry conditions, the increase in flow rates is found to decrease the detachment of hydrophobic polyethylene of two sizes and of hydrophilic polymethylmethacrylate. Intermittent flow is found to result in effluent peaks and a higher rate of MP detachment compared to steady flow. The ionic strength of inflow drops in a stepwise manner, leading to abrupt peaks followed by a tail corresponding to the arrival of each ionic strength front. Each step increase in flow rate leads to a steep peak followed by slow release over several pore volumes. Although transient flow facilitates the detachment of MPs, drying significantly impedes the detachment of MPs irrespective of flow regime. Ultraviolet weathering of MPs for 60 days weakens the inhibition effect of drying on hydrophilic polymethylmethacrylate, facilitating their detachment. Furthermore, the release of MPs decreases markedly with an increase in air-drying duration from 0 h to 72 h. Hydrus-1D two-site kinetic models are used to successfully simulate time-dependent processes, implying that drying heightens the energy barrier for MPs to detach. Our analysis confirms the significance of moisture in determining the remobilization of MPs, providing valuable insights concerning the fate of MPs in unsaturated substrate under prolonged drought conditions.

Foveal Microstructure and Visual Outcomes after Pars Plana Vitrectomy in Patients with Different Types of Epiretinal Membrane Foveoschisis.

INTRODUCTION: The aim of this study was to evaluate the clinical characteristics and surgical outcomes of the epiretinal membrane foveoschisis (ERM-FS) with different morphological types. METHODS: This retrospective observational study reviewed 44 consecutive ERM-FS patients who underwent ERM surgery. According to the optical coherence tomography images, ERM-FS was classified into three groups: group A, FS crossed the fovea with the foveola elevated; group B, FS located at the foveal edges with a near-normal central foveal point thickness; and group C, FS with undermined foveal edges with a near-normal central foveal point thickness. RESULTS: There were 10 eyes in group A, 20 eyes in group B, and 14 eyes in group C. Preoperatively, eyes in group A had the best best-corrected visual acuity (BCVA), the thickest central foveal point thickness, and the highest ellipsoid zone (EZ) intact rate among the three groups. After surgery, a resolution of foveoschisis was observed in 40.0%, 45.0%, and 50.0% of the eyes in group A, group B, and group C (p = 0.928), respectively. BCVA was significantly improved postoperatively. Although there was no significant difference in BCVA among the three groups at 1 month postoperatively, BCVA of group A was the best at 4 and 10 months. Correlation analysis indicated that the type of ERM-FS, baseline BCVA, central foveal point thickness, and postoperative EZ continuity (all p < 0.05) were important factors for the final BCVA. CONCLUSIONS: The damage to the retinal structure and visual function was milder in group A ERM-FS. Our study emphasized the necessity of OCT-based subtyping in patients with ERM-FS.

An Untethered Soft Crawling Robot Driven by Wireless Power Transfer Technology.

Soft robots based on flexible materials have attracted the attention due to high flexibility and great environmental adaptability. Among the common driving modes, electricity, light, and magnetism have the limitations of wiring, poor penetration capability, and sophisticated equipment, respectively. Here, an emerging wireless driving mode is proposed for the soft crawling robot based on wireless power transfer (WPT) technology. The receiving coil at the robot's tail, as an energy transfer station, receives energy from the transmitting coil and supplies the electrothermal responsiveness to drive the robot's crawling. By regulating the WPT's duration to control the friction between the robot and the ground, bidirectional crawling is realized. Furthermore, the receiving coil is also employed as a sensory organ to equip the robot with localization, ID recognition, and sensing capabilities based on electromagnetic coupling. This work provides an innovative and promising strategy for the design and integration of soft crawling robots, exhibiting great potential in the field of intelligent robots.

Transcription factor DgMYB recruits H3K4me3 methylase to DgPEROXIDASE to enhance chrysanthemum cold tolerance.

Cold affects the growth and development of plants. MYB transcription factors and histone H3K4me3 transferase ARABIDOPSIS TRITHORAXs (ATXs) play important regulatory functions in the process of plant resistance to low-temperature stress. In this study, DgMYB expression was responsive to low temperature, and overexpression of DgMYB led to increased tolerance, whereas the dgmyb mutant resulted in decreased tolerance of Chrysanthemum morifolium (Dendranthema grandiflorum var. Jinba) to cold stresses. Interestingly, we found that only peroxidase (POD) activity differed substantially between wild type (WT), overexpression lines, and the mutant line. A DgATX H3K4me3 methylase that interacts with DgMYB was isolated by further experiments. DgATX expression was also responsive to low temperature. Overexpression of DgATX led to increased tolerance, whereas the dgatx mutant resulted in decreased tolerance of chrysanthemum to cold stresses. Moreover, the dgmyb, dgatx, and dgmyb dgatx double mutants all led to reduced H3K4me3 levels at DgPOD, thus reducing DgPOD expression. Together, our results show that DgMYB interacts with DgATX, allowing DgATX to specifically target DgPOD, altering H3K4me3 levels, increasing DgPOD expression, and thereby reducing the accumulation of reactive oxygen species (ROS) in chrysanthemum.