Tangzukang Ointment (T-Ointment) is formulated by adding ingredients such as Astragalus, Angelica sinensis, Cinnamomi Cortex, and Coix seed to the traditional formula ErMiao Pill. This ointment has the effects of clearing heat and removing dampness, warming the yang energy, strengthening the qi, promoting blood circulation and tissue regeneration. Clinically, it is used by traditional Chinese medicine to treat diabetic foot ulcers (DFU). This study established an experimental rat model of ischemic DFU to investigate its therapeutic effect and potential mechanism in DFU. The results showed that T-Ointment significantly promoted wound healing in diabetic rats by inhibiting excessive inflammation, promoting epithelial regeneration, angiogenesis and collagen deposition. Network pharmacology analysis indicated that the core compounds (quercetin, phellochin, fumarine, poriferast-5-en-3beta-ol, and jaranol) in T-Ointment targeted multiple key genes (AKT1, TGF-β, HIF-1, TNF, IL-6, EGFR, TP53 and MAPK) and were closely associated with multiple important signaling pathways (PI3K/AKT, EGFR, TGF-β/Smad, and MAPK/ERK etc.). Analysis of combined results from KEGG pathway enrichment analysis and molecular docking, the key pathways potentially promoted by T-Ointment for diabetic foot ulcer healing were identified: the PI3K/AKT and EGFR pathways. Further research has confirmed that T-Ointment indeed reversed the downregulation of the PI3K/AKT and EGFR pathways in the diabetic foot ulcer group. This indicates that its role in accelerating ulcer healing is closely related to the activation of these two pathways. In conclusion, this study clarified the multi-component, multi-target and multi-pathway mechanism of T-Ointment in treating diabetic foot ulcers, providing a scientific basis for its clinical application.

Substrate-specific kinases catalyze addition of phosphate groups to specific amino acids, resulting in kinase-specific phosphorylation. It participates in various signaling pathways and regulation processes. The relevant computational methods can accelerate study of protein function research, disease exploration, and drug development. Existing approaches typically rely on global and local sequences to extract predictive features but often neglect position information and critical feature interaction, which is essential for effective feature representation. In this work, we propose a novel kinase-specific phosphorylation site prediction model, DCPPS, by leveraging dynamic embedding encoding and interaction between global and local representations. Specifically, to enrich sequence position information and strengthen features, we construct a dynamic embedding encoding (DEE) to capture amino acid semantics and positional information of upstream and downstream amino acids, dynamically optimizing feature embeddings. Considering the lack of in-depth feature interaction between local and global information, we design a cross-representation interaction unit (CRIU) to facilitate in-depth mining and complementary improvement of potential connections between multi-source features. Results of kinase-specific phosphorylation and multiple extended experiments show that DCPPS has better predictive performance and scalability. Further ablation studies demonstrate that incorporating global protein information, DEE, and CRIU markedly enhances phosphorylation site prediction accuracy, particularly in mitigating class imbalance.

BACKGROUND Numerous epidemiological studies have found that pesticide exposure is associated with the incidence of type 2 diabetes (T2D); however, the underlying mechanisms remain unknown. DNA methylation may play a role in this process. AIM To identify the genes associated with pesticide exposure and T2D by reviewing the current literature. METHODS We systematically searched PubMed and Embase for relevant studies that examined the association between pesticide exposure and DNA methylation, and studies on DNA methylation and T2D through January 15, 2024. RESULTS We identified six genes (Alu , CABLES1 , CDH1, PDX1 , PTEN , PTPRN2 ) related to pesticide exposure and T2D. We also suggested future research directions to better define the role of DNA methylation in the association between pesticide exposure and T2D. CONCLUSION DNA methylation of specific genes may play a vital role in the association between pesticide exposure and T2D.

BACKGROUND:Human spermatogonial stem cells (SSCs) exhibit a remarkable capacity for proliferation, crucial for sustaining spermatogenesis throughout life. While the Cullin-RING E3 ubiquitin ligase 2 (CRL2) complex is known to regulate various cellular functions, its precise role in human SSCs has not been fully elucidated. This study aimed to investigate a novel variant of the CRL2 complex, termed CRL2LRRC41, and its role in SSC function. METHODS:We utilized molecular biology techniques, including gene knockdown and functional assays, to assess the effects of CRL2LRRC41 on the proliferative and migratory abilities of human spermatogonial stem cell-like cell (SSCLC) line. Additionally, we employed proteomics and biochemical approaches to identify potential substrates of CRL2LRRC41. We specifically focused on ATP-dependent RNA helicase DDX5, a known regulator of spermatogenesis, to explore its interaction with CRL2LRRC41 and the downstream molecular mechanisms involved. RESULTS:Our findings revealed that the disruption or dysfunction of CRL2LRRC41 led to reduced proliferative and migratory abilities in human SSCLCs. Through our investigation, we identified DDX5 as a ubiquitination substrate of CRL2LRRC41. Notably, the ubiquitination of DDX5 fosters its interaction with the RNA-binding protein ELAVL1, without directing DDX5 towards degradation via the ubiquitin-proteasome system (UPS). This interaction enhances the stability of the downstream transcript, Noggin (NOG), thereby supporting human SSCLC proliferation and migration. CONCLUSIONS:This study provides the first identification of the CRL2LRRC41 complex in human SSCLCs and elucidates the molecular mechanisms by which CRL2LRRC41 facilitates SSCLC function via ubiquitination-mediated protein interactions. These findings offer novel insights into the molecular underpinnings of male infertility.

Ischemic stroke has a higher survival rate and is more likely to result in cognitive impairment than hemorrhagic stroke. The primary pathological mechanism underlying cognitive impairment involves dysfunction of neural circuits and damage to specific brain regions. This review aims to investigate the role of the hippocampus in cognitive impairment following a stroke. A review of the literature suggests that the hippocampus is a metabolically active structure that is easily involved in various metabolic states, such as hypoxia and hypoglycaemia. The functional changes in hippocampal cells associated with poststroke cognitive impairment mainly manifest as neuronal apoptosis, impaired synaptic plasticity, and decreased neurogenesis. The primary pathological mechanism of poststroke cognitive impairment involves a complex cascade of reactions, including neuroinflammatory activation, bursts of oxidative stress, and neuronal apoptosis induced by mitochondrial dysfunction. Interventional drugs for cognitive impairment after cerebral ischemia include neuroprotective drugs, traditional Chinese medicines and their extracts, and stem cell therapies. Many of these drugs have unique advantages, including the inhibition of neuroinflammation, the prevention of apoptosis, and the promotion of neurogenesis. They hold great potential for the prevention and treatment of cognitive impairment following cerebral ischemia. However, most current studies are animal experiments, and relatively few clinical studies exist. In future research, emphasis should be placed on interventions for cognitive impairment following cerebral ischemia. These findings offer novel perspectives for the treatment of cognitive impairment after cerebral ischemia. Finally, the role of hippocampal cell dysfunction in other diseases associated with cognitive decline is briefly discussed. The aim of this review is to provide researchers with a comprehensive overview of the role of the hippocampus in cognitive impairment and its intervention strategies.

The PDCA cycle, also known as the PDCA (Plan-Do-Check-Act) cycle, is a well-established continuous quality improvement framework. This study aimed to evaluate the impact of implementing a nursing safety management strategy grounded in the PDCA cycle and safety culture principles in the context of Day Ward chemotherapy. This a prospective group comparison study (cohort comparison) based on principles of randomization. A total of 120 patients receiving intravenous chemotherapy at the Day Ward of Nantong First People’s Hospital from January 2023 to December 2023 were recruited as research participants. They were randomly assigned to either a control group or a study group, which were managed using the conventional nursing quality management approach and the PDCA cycle-based safety culture management method, respectively. The primary outcomes measured were nursing satisfaction, chemotherapy-related symptom burden, and the incidence of total implantable venous access port catheter (TIVAP)-related adverse events. After three months, the study group showed significantly lower scores on all MSAS-SF subscales (GDI: 1.05 ± 0.33, PHYS: 0.69 ± 0.35, PSYCH: 1.15 ± 0.42, TMSAS: 2.62 ± 0.34) compared to the control group (GDI: 1.22 ± 0.47, PHYS: 0.85 ± 0.32, PSYCH: 1.43 ± 0.73, TMSAS: 2.81 ± 0.36) (all P < 0.05). Nursing satisfaction was higher in the study group (95.00

Ovarian clear cell carcinoma (OCCC) represents a rare and aggressive subtype of epithelial ovarian cancer with distinctive clinical and molecular characteristics. However, the identification, origin, and molecular features of the malignant epithelial cells in OCCC remain poorly studied. We establish an OCCC-associated transcriptional landscape using single-cell RNA sequencing and investigated the properties of epithelial cells in tissues from normal ovaries, ovarian endometriosis, primary OCCC and recurrent OCCC to assess the status of malignant epithelial cells. We identify a specific subcluster of malignant epithelial cells and further analyze them to discover 173 candidate factors associated with OCCC. Regulon and pseudotime trajectory analyses reveal six transcription factors (TFs) and their corresponding targets among these candidate factors, highlighting their roles in OCCC onset and reoccurrence. Through experimental validation, we confirm the crucial involvement of STAT3, KLF5, and TRIM28 in the proliferation and migration of OVISE cells. Silencing these three TFs also results in the down-regulation of their associated TF targets linked to OCCC. Overall, we characterize complex malignant-like cell populations at single-cell resolution and highlighted several TFs and their targets, providing essential resources for understanding the regulatory mechanisms underlying OCCC initiation and recurrence.

Neuroinflammation plays a key role in the progression of secondary brain injury after ischemic stroke, and exosomes have been increasingly recognized to eliminate inflammatory responses through various mechanisms. This study aimed to explore the effect and possible mechanism of human umbilical vein endothelial cells derived exosomes (H-EXOs) on neuroinflammation. We established a transient middle cerebral artery occlusion/reperfusion (tMCAO/R) in male rats and oxygen-glucose-deprivation/reoxygenation (OGD/R) model in cultured neurons to mimic secondary brain injury after ischemic stroke in vivo. H-EXOs were administered at the same time of reperfusion. Results showed that the production of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6, and the transcription factor Krüppel-like factor 14 (KLF14) were significantly increased both in rat brain tissue and cultured neural cells after ischemic-reperfusion (I/R) injury. H-EXOs treatment significantly improved the cultured cell viability, reduced infarct sizes, mitigated neurobehavioral defects, and alleviated the expression of pro-inflammatory cytokines compared with the control group, indicating that H-EXOs exerted anti-inflammatory effect against I/R injury. Further studies revealed that the anti-inflammatory effect of H-EXOs could be weakened by small-interfering RNA (siKLF4) transfection. KLF14 was a protective factor produced during cerebral ischemia-reperfusion injury. In conclusion, H-EXOs protect neurons from inflammation after I/R injury by enhancing KLF14 expression.

Male infertility, frequently driven by oxidative stress, impacts half of infertile couples globally. Despite its significance, the precise mechanisms governing this process remain elusive. In this study, we demonstrate that ASB1, the substrate recognition subunit of a ubiquitin ligase, is highly expressed in the mouse testis. Mice lacking the Asb1 gene exhibit severe fertility impairment, characterized by oligoasthenoteratozoospermia. Subsequent investigations unveiled that Asb1 knockout (Asb1-KO) mice encountered excessive oxidative stress and decreased hydrogen sulfide (H2S) levels in their testes, and severe sperm DNA damage. Notably, the compromised fertility and sperm quality in Asb1-KO mice was significantly ameliorated by administering NaHS, a H2S donor. Mechanistically, ASB1 interacts with ELOB to induce the instability of sulfide-quinone oxidoreductase (SQOR) by enhancing its K48-linked ubiquitination on residues K207 and K344, consequently triggering proteasomal degradation. This process is crucial for preserving H2S homeostasis and redox balance. Overall, our findings offer valuable insights into the role of ASB1 during spermiogenesis and propose H2S supplementation as a promising therapeutic approach for oxidative stress-related male infertility.

Resveratrol (Res) has long been discovered to have antioxidant effects to prevent such as oxidation, inflammation, neurodegeneration and age-related diseases. However, its poor water solubility, low bioavailability and instability have become a barrier to its pharmaceutical application. In order to improve the neuroprotective effects and develop more potential usage of Res, three Res derivatives containing one or two glucose groups, i.e., Res-Glu1, Res-Glu2 and Res-Glu3, were designed and synthesized through click reaction. Res-Glu1, Res-Glu2 and Res-Glu3 were tested being better water solubility and stability compared to Res. Res derivatives reduced •OH radicals-induced DNA damage. PC12 assays indicated that glucosylated Res derivatives could alleviate H2O2-induced neurotoxicity and reduce intracellular ROS generation, demonstrating their neuroprotective effects. In addition, Res derivatives enhanced the protective effects on cerebral ischemia-reperfusion injury in rats. Res-Glu3 displayed the best neuroprotective effects among the three derivatives.

Testicular ageing is accompanied by a series of morphological changes, while the features of mitochondrial dysfunction remain largely unknown. Herein, we observed a range of age-related modifications in testicular morphology and spermatogenic cells, and conducted single-cell RNA sequencing on young and old testes in Drosophila. Pseudotime trajectory revealed significant changes in germline subpopulations during ageing. Our examination unveiled that genes showing bias in spermatids exhibited higher dN/dS than those in GSCs_Spermatogonia. Genes biased towards young GSCs_Spermatogonia displayed higher dN/dS than those in old GSCs_Spermatogonia. Interestingly, genes biased towards young spermatids demonstrated lower dN/dS in contrast to those in old spermatids, revealing the complexity of evolutionary adaptations during ageing. Furthermore, mitochondria associated events, including oxidative phosphorylation, TCA cycle and pyruvate metabolism, were significantly enriched in germline subpopulations. Specifically, mitochondrial function was significantly impaired during the process of testicular ageing, concurrently emphasising the role of several key nuclear genome-encoded mitochondrial regulatory genes, such as Hsp60B, fzo, Tim17b1 and mRpL12. Our data offer insights into testicular homeostasis regulated by mitochondrial function during the ageing process.

OBJECTIVE:To evaluate the efficacy and safety of Ginkgolide Meglumine Injection (GMI) combined with Butylphthalide in the treatment of Acute Ischemic Stroke (AIS), and provide reference for rational clinical medication. METHODS:PubMed, Embase, Web of science, CNKI, Wanfang, VIP and other databases were searched for published studies on the treatment of AIS with GMI combined with Butylphthalide in both Chinese and English. The search period was from the establishment of the database to July 2023. The included studies that met the inclusion criteria were analyzed using RevMan 5.3 software for Meta-analysis. RESULTS:A total of 25 studies involving 2362 patients (experimental group = 1182, control group = 1180) were included. The results of meta-analysis showed that the overall effective rate of the experimental group was significantly higher than that of the control group [RR = 1.21, 95% CI (1.16, 1.26), P< 0.00001]. In addition, compared with the control group, the experimental group showed significant improvement in NIHSS score [SMD = -1.59, % CI (-2.00-1.18), P< 0.00001] and ADL score [SMD = 2.12, 95% CI (1.52, -2.72), P<0.00001], significant decrease in CRP [SMD = -2.24, 95% CI (-3.31, -1.18), P<0.0001] and TNF-α [SMD = -2.74, 95% CI (-4.45, -1.03), P< 0.005] levels, and improvement in plasma viscosity [SMD = -0.86, 95% CI (-1.07, -0.66), P< 0.00001]. However, the influence on homocysteine level remains inconclusive. Furthermore, there was no significant difference in the incidence of adverse reactions between the two groups [SMD = 0.95, 95% CI (0.71, 1.28), P> 0.05]. CONCLUSION:GMI combined with Butylphthalide shows good clinical application effects and good safety in the treatment of AIS. However, more large-sample, multicenter, randomized controlled are needed to confirm these findings.

As time goes by, the morbidity of diabetes mellitus continues to rise, and the economic burden of diabetic foot ulcers as a common and serious complication of diabetes is increasing. However, currently there is no unified clinical treatment strategy for this complication, and the therapeutic efficacy is unsatisfactory. Recent studies have revealed that biological effects of exosomes involved in multiple stages of the process of wound closure are similar to source cells. Compared with source cells, exosomes possess lowly immunogenicity, highly stability and easily stored, etc. Accumulating evidence confirmed that exosomes promote diabetic wound healing through various pathways such as promoting angiogenesis, collagen fiber deposition, and inhibiting inflammation. The superior therapeutic efficacy of exosomes in accelerating diabetic cutaneous wound healing has attracted an increasing attention. Notably, the molecular mechanisms of exosomes vary among different sources in the chronic wound closure of diabetes. This review focuses on the specific roles and mechanisms of different cell- or tissue-derived exosomes relevant to wound healing. Additionally, the paper provides an overview of the current pre-clinical and clinical applications of exosomes, illustrates their special advantages in wound repair. Furthermore, we discuss the potential obstacles and various solutions for future research on exosomes in the management of diabetic foot ulcer. The aim is to offer novel insights and approaches for the treatment of diabetic foot ulcer.

During spermatogenesis, mitochondria extend along the whole length of spermatid tail and offer a structural platform for microtubule reorganization and synchronized spermatid individualization, that eventually helps to generate mature sperm in Drosophila. However, the regulatory mechanism of spermatid mitochondria during elongation remains largely unknown. Herein, we demonstrated that NADH dehydrogenase (ubiquinone) 42 kDa subunit (ND-42) was essential for male fertility and spermatid elongation in Drosophila. Moreover, ND-42 depletion led to mitochondrial disorders in Drosophila testes. Based on single-cell RNA-sequencing (scRNA-seq), we identified 15 distinct cell clusters, including several unanticipated transitional subpopulations or differentiative stages for testicular germ cell complexity in Drosophila testes. Enrichments of the transcriptional regulatory network in the late-stage cell populations revealed key roles of ND-42 in mitochondria and its related biological processes during spermatid elongation. Notably, we demonstrated that ND-42 depletion led to maintenance defects of the major mitochondrial derivative and the minor mitochondrial derivative by affecting mitochondrial membrane potential and mitochondrial-encoded genes. Our study proposes a novel regulatory mechanism of ND-42 for spermatid mitochondrial derivative maintenance, contributing to a better understanding of spermatid elongation.

The toxic heavy metal antimony (Sb) is ubiquitous in our daily lives. Various models have shown that Sb induces neuronal and reproductive toxicity. However, little is known about the developmental toxicity of Sb exposure during gestation and the underlying mechanisms. To study its effects on growth and development, Drosophila stages from eggs to pupae were exposed to different Sb concentrations (0, 0.3, 0.6 and 1.2 mg/mL Sb); RNA sequencing was used to identify the underlying mechanism. The model revealed that prenatal Sb exposure significantly reduced larval body size and weight, the pupation and eclosion rates, and the number of flies at all stages. With 1.2 mg/mL Sb exposure in 3rd instar larvae, 484 genes were upregulated and 694 downregulated compared to controls. Biological analysis showed that the disrupted transcripts were related to the oxidative stress pathway, as verified by reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) and glutathione (GSH) intervention experiments. Sb exposure induced oxidative stress imbalance could be rectified by chelation and antioxidant effects of NAC/GSH. The Drosophila Schneider 2 (S2) model further demonstrated that NAC and GSH greatly ameliorated cell death induced by Sb exposure. In conclusion, gestational Sb exposure disrupted oxidative stress homeostasis, thereby impairing growth and development.

OBJECTIVE:Our previous study found that ErbB4 gene expression was changed after oxygen-glucose deprivation/reperfusion (OGD/R). However, the exact role and mechanism of ErbB4 in brain ischemia are largely unknown. In this study, we explored the protective effects of ErbB4 and its possible mechanism after OGD/R. METHODS:Cerebral ischemia/reperfusion (I/R) injury model was established in vitro and in vivo. Cell viability, apoptosis, and ROS production were measured by MTT, TUNEL, and fluorescent probe 2', 7'-dichlorofluorescein diacetate (DCFH-DA). Infarct size was evaluated by TTC. We performed bioinformatics analyses to screen for novel key genes involved in ErbB4 changes. RNA-Seq was used to transcriptome analysis. RNA and protein expression were detected by quantitative RT‒PCR and western bloting. RESULTS:The expression of 80-kDa ErbB4 decreased after cerebral I/R injury in vitro and in vivo. Co-expression network analysis revealed that ErbB4 expression was correlated with the changes in Adrb1, Adrb2, Ldlr, and Dab2. Quantitative RT‒PCR revealed that the mRNA expression levels of Adrb1, Adrb2, and Dab2 were upregulated, and that of Ldlr was decreased after OGD/R. Activation of ErbB4 expression by neuregulin 1 (NRG1) significantly promoted cell survival, attenuated hippocampal apoptosis, and decreased ROS production after OGD/R. Furthermore, the elimination of ErbB4 using a specific siRNA reversed these beneficial effects. CONCLUSION:Our data revealed the neuroprotective effects of ErbB4 against OGD/R injury, and the action could be related to changes in the ErbB4 membrane-associated fragment and the expression of Adrb1, Adrb2, Ldlr, and Dab2.

Dear Editor, R-spondin3(RSPO3)is essential for vascular development and angiogenesis.Analyzing RSPO3-knockout embryos revealed severe vascular defects in the placenta(Aoki et al.2007).In both Xenopus and murine embryos,RSPO3 KO led to signifi-cant vascular defects(Kazanskaya et al.2008)and embryonic death(Kazanskaya et al.2008).In the placenta,RSPO3 could promote vascular endothelial growth factor(VEGF)expression(Kazanskaya et al.2008).RSPO3 is a ligand of low-density lipopro-tein receptor-related protein 6(LRP6)and leucine-rich repeat G protein-coupled receptor 4(LGR4)to form a multiple ligands-re-ceptors-cluster with Wnt and frizzled(FZD),thereby activating and amplifying downstream β-catenin signaling(Jin and Yoon 2012;Tocci et al.2020).RSPO3 neutralizes two trans-membrane E3 ubiquitin ligases,zinc and ring finger 3(ZNRF3)/ring finger pro-tein 43(RNF43).The two could decrease cell-surface Wnt recep-tors(Jin and Yoon 2012;Tocci et al.2020).

This study serves to investigate the effects of the Smad pathway on TGFβ1-mediated RhoGDI expression and its binding to RhoGTPases in myofibroblast transdifferentiation. Myofibroblast transdifferentiation was induced by TGFβ1 in vitro. Cells were pretreated with different siRNAs or inhibitors. Myofibroblast transdifferentiation was detected by immunohistochemistry. Immunofluorescence was used to observe the nuclear translocation of Smad4, and PSR (Picrositius Red) staining was used to measure collagen concentration. TGFβ1 induced the phosphorylation of Smad2/3 and the nuclear translocation of Smad4 in human aortic adventitial fibroblasts (HAAFs). Furthermore, TGFβ1 increased the expression of RhoGDI and its binding to RhoGTPases. Nevertheless, inhibition of Smad2/3 phosphorylation decreased TGFβ1-induced RhoGDI1/2 expressions and RhoGDI2-RhoGTPases interactions. These data suggested that the inhibition of Smad phosphorylation attenuates myofibroblast transdifferentiation by inhibiting TGFβ1-induced RhoGDI1/2 expressions and RhoGDI-RhoGTPases signaling.

Autophagy has been implicated in stroke. Our previous study showed that the FoxO3 transcription factor promotes autophagy after transient cerebral ischemia/reperfusion (I/R). However, whether the Akt/FoxO3 signaling pathway plays a regulatory role in autophagy in cerebral I/R-induced oxidative stress injury is still unclear. The present study aims to investigate the effects of the Akt/FoxO3 signaling pathway on autophagy activation and neuronal injury in vitro and in vivo. By employing LY294002 or insulin to regulate the Akt/FoxO3 signaling pathway, we found that insulin pretreatment increased cell viability, decreased reactive oxygen species production, and enhanced the expression of antiapoptotic and autophagy-related proteins following H2O2 injury in HT22 cells. In addition, insulin significantly decreased neurological deficit scores and infarct volume and increased the expression of antiapoptotic and autophagy-related proteins following I/R injury in rats. However, LY294002 showed the opposite effects under these conditions. Altogether, these results indicate that Akt/FoxO3 signaling pathway activation inhibited oxidative stress-mediated cell death through activation of autophagy. Our study supports a critical role for the Akt/FoxO3 signaling pathway in autophagy activation in stroke.

Autophagy has been implicated in neurodegenerative diseases. Forkhead box O3 (FoxO3) transcription factors promote autophagy in heart and inhibit oxidative damage. Here we investigate the role of FoxO3 transcription factors in regulating autophagy after oxidative stress injury in immortalized mouse hippocampal cell line (HT22). The present study confirms that hydrogen peroxide (H2O2) injury could induce autophagy and FoxO3 activation in HT22 cells. In addition, overexpression of FoxO3 enhanced H2O2-induced autophagy activation and suppressed neuronal cell damage, while knockdown of FoxO3 reduced H2O2-induced autophagy activation and exacerbated neuronal cell injury. Inhibition of autophagy by 3-methyladenine (3-MA) resulted in reduced cell viability, increased production of reactive oxygen species (ROS), promoted nuclear condensation, and decreased expression of antiapoptotic and autophagy-related proteins, indicating that autophagy may have protective effects on H2O2-induced injury in HT22 cells. Moreover, overexpression of FoxO3 prevented exacerbation of brain damage induced by 3-MA. Taken together, these results show that activation of FoxO3 could induce autophagy and inhibit H2O2-induced damage in HT22 cells. Our study demonstrates the critical role of FoxO3 in regulating autophagy in brain.