Direct Access to 3,4,6-Trisubstituted 2-Pyrones via Carbene Catalysis.

An N-heterocyclic carbene-catalyzed [4 + 2] annulation of ß,γ-unsaturated α-keto esters and phenylacetate esters was developed for the direct and efficient construction of 2-pyrones. This approach provides a practical synthesis pathway for various 3,4,6-trisubstituted 2-pyrones in moderate to good yields and features broad substrate scope and good functional group tolerance. Moreover, the products can also be readily transformed to naphthalene and acylamide.

Carbene-Catalyzed [3 + 3] Annulation of Enals and Vinyl Sulfoxonium Ylides.

Carbene-catalyzed [3 + 3] annulation of enals and vinyl sulfoxonium ylides has been demonstrated. This method efficiently synthesizes a range of 2-sulfenylidene-3-cyclohexen-1-ones with high atom economy. Notably, the presence of the sulfoxonium ylide moiety in the obtained products significantly enhances their potential for further synthetic transformations.

Laboratory Investigation of Dodecylbenzene Detection in Seawater Via Fluorometric Index of Excitation-emission Matrix Spectra.

​Oil-filled submarine cables have come to dominate the reliable delivery of electrical power today, but oil leakages still remain a challenge. This work focuses on exploring the detection of dodecylbenzene (DDB) after leakage. DDB can be dispersed as small droplets in seawater, which the diameter of DDB droplets increased from 60 nm to 200 nm with the increase of the concentration from 10 ppm to 1000 ppm. Moreover, excitation-emission matrix (EEM) spectra were employed to investigated the detection of DDB. DDB has characteristic fluorescence peaks located at λEx/λEm = 225/350 and 255/350. The FIo/w value of 50 ppm DDB was 28.869, which was significantly different from simulated seawater low as 1.3926. Contaminants such as metal ions (Cu2+, Cd2+, Zn2+) and organic matter (tetracycline) do not significantly affect the qualitative identification of DDB by EEMs. This study suggests a research basis for exploring the way for detection of DDB.

Rapid screening and evaluation of natural antioxidants from leaf, stem, and root of Artemisia argyi by online liquid microextraction combined with HPLC-based antioxidant assay system coupled with calibration quantitative analysis.

To reveal the utilization value of leaf, stem, and root of Artemisia argyi, a rapid online liquid microextraction combined with a high-performance liquid chromatography coupled with 2,2-nitrogen-di (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt antioxidant assay system was established for analysis of antioxidants in the leaf, stem, and root of A. argyi, and a calibration quantitative method of antioxidant activity with equivalent chlorogenic acid was proposed. Thirty-three positive peaks were identified; among them, 12 compounds were found that possess good antioxidant activity including eleven organic acids (components 2-4, 8, 11-14, 17, 19, and 21) and one flavonoids (component 22). The proposed calibration quantitative method avoided the influence of content of compound and compared the extent of radical scavenging capacity of five antioxidant compounds, which were ranked as follow: 3,5-dicaffeoylquinic acid > 3,4-dicaffeoylquinic acid ≈ 4,5-dicaffeoylquinic acid > 1,4-dicaffeoylquinic acid > chlorogenic acid. In conclusion, this study provided composition and biological potential for the future development of the leaf, stem, and root of A. argyi. It is believed that the online liquid microextraction combined with high-performance liquid chromatography based antioxidant assay system can be widely used for the rapid screening of natural antioxidant components in the different parts of natural products.

Analysis of Antioxidants from Three Parts of Polygonum Chinense Based on Online Extraction High Performance Liquid Chromatography Antioxidant Analysis System.

In the current study, a rapid online extraction combined with high-performance liquid chromatography (HPLC) antioxidant analysis approach was developed to explore the antioxidant ingredients in three different parts of Polygonum Chinense. A total of 22 chromatographic peaks were found, among which 8 components were found for the first time. And among these 22 peaks, 19 of them were demonstrated as antioxidants. Furthermo, all three parts from Polygonum Chinense exhibited antioxidant activity (13.19, 3.89, and 19.85 mg CE/g extract, for the leaf, stem, and flower, respectively) and the antioxidant activity values of the leaf and flower were more than 3 times that of the stem. The leaf, flower, and stem of Polygonum Chinense had 17, 13, and 15 antioxidant components, respectively. Among them, neochlorogenic acid (2) and quercetin-3-O-malonylglucoside (18) were considered as the main antioxidants in the leaf of Polygonum Chinense. In terms of flower, geraniin (10) and quercetin-3-O-malonylglucoside (18) were proved as the major antioxidants. This is the first report on the antioxidant components from different parts of Polygonum Chinense. It provides foundational scientific data for the continued exploration of chemical and pharmacological study of Polygonum Chinense, beneficial for the future product development and quality evaluation improvement of Polygonum Chinense.

Current understanding on antibacterial mechanisms and research progress of tea polyphenols as a supplementary disinfectant for drinking water.

Disinfection by-products (DBPs) generated during the disinfection of drinking water have become an urgent problem. So, tea polyphenol, a natural green disinfectant, has attracted widespread attention in recent years. This review summarizes the antibacterial mechanism of tea polyphenols and the recent findings on tea polyphenols as disinfectants for drinking water. These studies show that tea polyphenol is an antibacterial agent that works through different mechanisms and can be used as a supplementary disinfectant because of its higher lasting effect and economical cost. The dosage of tea polyphenols as a disinfectant of ultrafiltration effluent is the lowest among all the tea polyphenols disinfection methods, which can ensure the microbial safety of drinking water. This application of tea polyphenols is deemed a practical solution to solving the issue of disinfecting drinking water and reducing DBPs. However, it is necessary to further explore the influence of factors such as pipeline materials on the disinfection process and efficacy to expand the application scope of tea polyphenols. The large-scale application of tea polyphenols still needs to be fine-tuned but with new developments in tea polyphenol purification technology and the long-term need for drinking water that is safe for human consumption, tea polyphenols have good prospects for further development.

Reassessment of the Effect of Transcutaneous Auricular Vagus Nerve Stimulation Using a Novel Burst Paradigm on Cardiac Autonomic Function in Healthy Young Adults.

BACKGROUND: Transcutaneous auricular vagus nerve stimulation (taVNS) may modulate cardiac autonomic function. However, the response rate of the traditional tonic paradigm is low, and the results remain inconsistent. A recent pilot study presented a novel burst paradigm to activate the cardiac parasympathetic system, which might offer a new approach to treat cardiac autonomic function. The present study reassessed the effect of burst taVNS on modulating heart rate variability and explored the difference between burst and traditional tonic paradigms. MATERIALS AND METHODS: Forty-two young adults were recruited for this study. Each participant underwent three types of taVNS with sham (30 sec of stimulation), tonic (25 Hz, 500 µsec), and burst (five pulses at 500 Hz every 200 msec) paradigms, respectively, with simultaneous electrocardiogram recording. One-way analysis of variance, multivariate analysis of variance, and linear regression were used for analysis. Multiple testing was performed using Bonferroni correction. RESULTS: Both burst and tonic paradigms induced a significant decrease in heart rate, which continued until poststimulation, and increased cardiac parasympathetic activity. Moreover, two parasympathetic system indicators showed significant increase only in burst taVNS. The response rates during burst (35.7%) and tonic (38.1%) stimulations were both higher than that during sham stimulation (11.9%). The response to taVNS showed parameter specificity with few nonresponders to the tonic paradigm responding to the burst paradigm. The overall response rate increased from 38.1% in tonic taVNS to 54.8% in taVNS using both burst and tonic paradigms. For both burst and tonic responders, baseline cardiac parasympathetic activity was found to be significantly negatively correlated with changes during stimulation. CONCLUSION: The burst parameter could be used as an alternative strategy for regulating cardiac parasympathetic function by taVNS, which has the potential to be used as a complementary paradigm to traditional tonic taVNS for promoting clinical treatment efficacy.

Magnetization transfer in liposome and proteoliposome samples that mimic the protein and lipid composition of myelin.

Although magnetization transfer (MT) has been widely used in brain MRI, for example in brain inflammation and multiple sclerosis, the detailed molecular origin of MT effects and the role that proteins play in MT remain unclear. In this work, a proteoliposome model system was used to mimic the myelin environment and to examine the roles of protein, cholesterol, brain cerebrosides, and sphingomyelin embedded in the liposome matrix. Exchange parameters were determined using a double-quantum filter experiment. The goal was to determine the relative contributions to exchange and MT of cerebrosides, sphingomyelin, cholesterol, and proteins in 1,2-dimyristoyl-sn-glycero-3-phosphocholine bilayers. The main finding was that cerebrosides produced the strongest exchange effects, and that these were even more pronounced than those found for proteins. Sphingomyelin (which also has exchangeable groups at the head of the fatty acid chains, albeit closer to the lipid acyl chains) and cholesterol showed only minimal transfer. Overall, the extracted exchange rates appeared much smaller than commonly assumed for -OH and -NH groups.

A Feature Extraction Method Based on Differential Entropy and Linear Discriminant Analysis for Emotion Recognition.

Feature extraction of electroencephalography (EEG) signals plays a significant role in the wearable computing field. Due to the practical applications of EEG emotion calculation, researchers often use edge calculation to reduce data transmission times, however, as EEG involves a large amount of data, determining how to effectively extract features and reduce the amount of calculation is still the focus of abundant research. Researchers have proposed many EEG feature extraction methods. However, these methods have problems such as high time complexity and insufficient precision. The main purpose of this paper is to introduce an innovative method for obtaining reliable distinguishing features from EEG signals. This feature extraction method combines differential entropy with Linear Discriminant Analysis (LDA) that can be applied in feature extraction of emotional EEG signals. We use a three-category sentiment EEG dataset to conduct experiments. The experimental results show that the proposed feature extraction method can significantly improve the performance of the EEG classification: Compared with the result of the original dataset, the average accuracy increases by 68%, which is 7% higher than the result obtained when only using differential entropy in feature extraction. The total execution time shows that the proposed method has a lower time complexity.

Modelling Thermally Induced Non-Equilibrium Gas Flows by Coupling Kinetic and Extended Thermodynamic Methods.

Thermally induced non-equilibrium gas flows have been simulated in the present study by coupling kinetic and extended thermodynamic methods. Three different types of thermally induced gas flows, including temperature-discontinuity- and temperature-gradient-induced flows and radiometric flow, have been explored in the transition regime. The temperature-discontinuity-induced flow case has shown that as the Knudsen number increases, the regularised 26 (R26) moment equation system will gradually loss its accuracy and validation. A coupling macro- and microscopic approach is employed to overcome these problems. The R26 moment equations are used at the macroscopic level for the bulk flow region, while the kinetic equation associated with the discrete velocity method (DVM) is applied to describe the gas close to the wall at the microscopic level, which yields a hybrid DVM/R26 approach. The numerical results have shown that the hybrid DVM/R26 method can be faithfully used for the thermally induced non-equilibrium flows. The proposed scheme not only improves the accuracy of the results in comparison with the R26 equations, but also extends their capability with a wider range of Knudsen numbers. In addition, the hybrid scheme is able to reduce the computational memory and time cost compared to the DVM.

SKIP controls flowering time via the alternative splicing of SEF pre-mRNA in Arabidopsis.

BACKGROUND: Similar to other eukaryotes, splicing is emerging as an important process affecting development and stress tolerance in plants. Ski-interacting protein (SKIP), a splicing factor, is essential for circadian clock function and abiotic stress tolerance; however, the mechanisms whereby it regulates flowering time are unknown. RESULTS: In this study, we found that SKIP is required for the splicing of serrated leaves and early flowering (SEF) pre-messenger RNA (mRNA), which encodes a component of the ATP-dependent SWR1 chromatin remodeling complex (SWR1-C). Defects in the splicing of SEF pre-mRNA reduced H2A.Z enrichment at FLC, MAF4, and MAF5, suppressed the expression of these genes, and produced an early flowering phenotype in skip-1 plants. CONCLUSIONS: Our findings indicate that SKIP regulates SWR1-C function via alternative splicing to control the floral transition in Arabidopsis thaliana.

Study on the inactivation effect and mechanism of EGCG disinfectant on Bacillus subtilis.

The widespread use of chlorine-based disinfectants in drinking water treatment has led to the proliferation of chlorine-resistant bacteria and the risk of disinfection byproducts (DBPs), posing a serious threat to public health. This study aims to explore the effectiveness and potential applications of epigallocatechin gallate (EGCG) against chlorine-resistant Bacillus and its spores in water, providing new insights for the control of chlorine-resistant bacteria and improving the biological stability of distribution systems. The inactivation effects of EGCG on Bacillus subtilis (B. subtilis) and its spores were investigated using transmission electron microscopy, ATP measurement, and transcriptome sequencing analysis to determine changes in surface structure, energy metabolism, and gene expression levels, thereby elucidating the inactivation mechanism. The results demonstrate the potential application of EGCG in continuously inhibiting chlorine-resistant B. subtilis in water, effectively improving the biological stability of the distribution system. However, EGCG is not suitable for treating raw water with high spore content and is more suitable as a supplementary disinfectant for processes with strong spore removal capabilities, such as ozone, ultraviolet, or ultrafiltration. EGCG exhibits a disruptive effect on the morphological structure and energy metabolism of B. subtilis and suppresses the synthesis of substances, energy metabolism, and normal operation of the antioxidant system by inhibiting the expression of multiple genes, thereby achieving the inactivation of B. subtilis.

Facile construction of 2-pyrones under carbene catalysis.

2-Pyrones are valuable structural motifs in organic chemistry, found in numerous natural products and pharmaceuticals. The synthesis of these heterocycles has been significantly advanced by the application of N-Heterocyclic Carbene (NHC) catalysis. This review examines the recent advancements in NHC-catalyzed synthesis of 2-pyrones, highlighting key methodologies, mechanisms, and synthetic applications. NHC catalysis has revolutionized the synthesis of 2-pyrones, providing efficient, selective, and versatile methods for constructing these valuable heterocycles.

Diabetes Mellitus Inhibits Hair Follicle Regeneration by Inducing Macrophage Reprogramming-Mediated Pyroptosis.

Background: Diabetes mellitus (DM) is known to inhibit skin self-renewal and hair follicle stem cell (HFSC) activation, which may be key in the formation of chronic diabetic wounds. This study aimed to investigate the reasons behind the suppression of HFSC activation in DM mice. Methods: Type 1 DM (T1DM) was induced in 6-week-old mice via streptozotocin, and hair follicle growth was subsequently monitored. RNA sequencing, bioinformatics analyses, qRT‒PCR, immunostaining, and cellular experiments were carried out to investigate the underlying mechanisms involved. Results: T1DM inhibited HFSC activation, which correlated with an increase in caspase-dependent programmed cell death. Additionally, T1DM triggered apoptosis and pyroptosis, predominantly in HFSCs and epidermal regions, with pyroptosis being more pronounced in the inner root sheath of hair follicles. Notably, significant cutaneous immune imbalances were observed, particularly in macrophages. Cellular experiments demonstrated that M1 macrophages inhibited HaCaT cell proliferation and induced cell death, whereas high-glucose environments alone did not have the same effect. Conclusion: T1DM inhibits HFSC activation via macrophage reprogramming-mediated caspase-dependent pyroptosis, and there is a significant regional characterization of cell death. Moreover, T1DM-induced programmed cell death in the skin may be more closely related to immune homeostasis imbalance than to hyperglycemia itself. These findings shed light on the pathogenesis of diabetic ulcers and provide a theoretical basis for the use of hair follicle grafts in wound repair.

ROS-mediated lysosomal membrane permeabilization and autophagy inhibition regulate bleomycin-induced cellular senescence.

Bleomycin exhibits effective chemotherapeutic activity against multiple types of tumors, and also induces various side effects, such as pulmonary fibrosis and neuronal defects, which limit the clinical application of this drug. Macroautophagy/autophagy has been recently reported to be involved in the functions of bleomycin, and yet the mechanisms of their crosstalk remain insufficiently understood. Here, we demonstrated that reactive oxygen species (ROS) produced during bleomycin activation hampered autophagy flux by inducing lysosomal membrane permeabilization (LMP) and obstructing lysosomal degradation. Exhaustion of ROS with N-acetylcysteine relieved LMP and autophagy defects. Notably, we observed that LMP and autophagy blockage preceded the emergence of cellular senescence during bleomycin treatment. In addition, promoting or inhibiting autophagy-lysosome degradation alleviated or exacerbated the phenotypes of senescence, respectively. This suggests the alternation of autophagy activity is more a regulatory mechanism than a consequence of bleomycin-induced cellular senescence. Taken together, we reveal a specific role of bleomycin-induced ROS in mediating defects of autophagic degradation and further regulating cellular senescence in vitro and in vivo. Our findings, conversely, indicate the autophagy-lysosome degradation pathway as a target for modulating the functions of bleomycin. These provide a new perspective for optimizing bleomycin as a clinically applicable chemotherapeutics devoid of severe side-effects.Abbreviations: AT2 cells: type II alveolar epithelial cells; ATG7: autophagy related 7; bEnd.3: mouse brain microvascular endothelial cells; BNIP3L: BCL2/adenovirus E1B interacting protein 3-like; CCL2: C-C motif chemokine ligand 2; CDKN1A: cyclin dependent kinase inhibitor 1A; CDKN2A: cyclin dependent kinase inhibitor 2A; FTH1: ferritin heavy polypeptide 1; γ-H2AX: phosphorylated H2A.X variant histone; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HUVEC: human umbilical vein endothelial cells; HT22: hippocampal neuronal cell lines; Il: interleukin; LAMP: lysosomal-associated membrane protein; LMP: lysosome membrane permeabilization; MTORC1: mechanistic target of rapamycin kinase complex 1; NAC: N-acetylcysteine; NCOA4: nuclear receptor coactivator 4; PI3K: phosphoinositide 3-kinase; ROS: reactive oxygen species; RPS6KB/S6K: ribosomal protein S6 kinase; SA-GLB1/ß-gal: senescence-associated galactosidase, beta 1; SAHF: senescence-associated heterochromatic foci; SASP: senescence-associated secretory phenotype; SEC62: SEC62 homolog, preprotein translocation; SEP: superecliptic pHluorin; SQSTM1/p62: sequestosome 1; TFEB: transcription factor EB.

Preparation, evaluation and application of MRI detectable sunitinib-loaded calcium alginate/poly(acrylic acid) hydrogel microspheres.

Transcatheter arterial chemoembolization (TACE) is an effective method for the treatment of unresectable hepatocellular carcinoma. Although many embolic agents have been developed in TACE, there are few ideal embolic agents that combine drug loading, imaging properties and vessel embolization. Here, we developed novel magnetic embolic microspheres that could simultaneously load sunitinib malate (SU), be detected by magnetic resonance imaging (MRI) and block blood vessels. Calcium alginate/poly (acrylic acid) hydrogel microspheres (CA/PAA-MDMs) with superparamagnetic iron oxide nanoparticles (SPIONs) modified by citric acid were prepared by a drip and photopolymerization method. The embolization and imaging properties of CA/PAA-MDMs were evaluated through a series of experiments such as morphology, X-ray diffraction and X-ray photoelectron spectroscopy, magnetic responsiveness analysis, elasticity, cytotoxicity, hemolysis test, in vitro MRI evaluation, rabbit ear embolization and histopathology. In addition, the ability of drug loading and drug release of CA/PAA-MDMs were investigated by using sunitinib (SU) as the model drug. In conclusion, CA/PAA-MDMs showed outstanding drug loading capability, excellent imaging property and embolization effect, which would be expected to be used as a potential biodegradable embolic agent in the clinical interventional therapy.

The LHT Gene Family in Rice: Molecular Characterization, Transport Functions and Expression Analysis.

Amino acid transporters (AATs) are integral membrane proteins and play important roles in plant growth and development as well as environmental responses. In contrast to the amino acid permease (AAP) subfamily, functional studies of the lysine and histidine transporter (LHT) subfamily have not been made in rice. In the current study, six LHT genes were found in the rice genome. To further investigate the functions of these genes, analyses were performed regarding gene and protein structures, chromosomal locations, evolutionary relationships, cis-acting elements of promoters, gene expression, and yeast complementation. We found that the six OsLHT genes are distributed on 4 out of the 12 chromosomes and that the six OsLHT genes were grouped into two clusters based on the phylogenetic analysis. Protein structure analyses showed that each OsLHT protein has 11 helical transmembrane domains. Yeast complementation assays showed that these OsLHT genes have conserved transport substrates within each cluster. The four members from cluster 1 showed broad amino acid selectivity, while OsLHT5 and OsLHT6 may transport other substrates besides amino acids. Additionally, quantitative real-time PCR analysis of the six OsLHT genes revealed that they have different expression patterns at different developmental stages and in different tissues. It also revealed that some OsLHT genes were responsive to PEG, NaCl and cold treatments, indicating their critical roles in abiotic stress response. Our results will be useful for further characterizing the crucial biological functions of rice LHT genes.

Online Microextraction Coupled with HPLC-ABTS for Rapid Analysis of Antioxidants from the Root of Polygonum bistorta.

The root of Polygonum bistorta (PB) is a traditional Chinese medicinal plant material widely used in China. It has been commonly used for the treatment of hemostasis, detumescence, diarrhea, snake bite, and acute gastroenteritis. However, the research on the antioxidant properties and bioactive compounds from PB is inadequate. In the current research, an online microextraction (OLME) coupled with a high-performance liquid chromatography coupled with the 2,2-nitrogen-di (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt antioxidant assay (HPLC-ABTS) system for rapid analysis of antioxidants from PB was proposed. The PB sample (0.17 mg) was online extracted by mobile phase (acetonitrile and 0.2% acetic acid); a Poroshell 120 SB-Aq column was used for separation; then, an online ABTS assay system was used for screening the antioxidants. Finally, ten components were found in PB, and among them, eight components possessed antioxidant activities. Furthermore, five components (gallic acid, neochlorogenic acid, caffeic acid, chlorogenic acid, and an unknown compound) were proved as major antioxidants when compared with rutin as an antioxidant marker. The results showed that the developed OLME-HPLC-ABTS system was a simple, rapid, green, and efficient instrument for the screening of antioxidants from PB, which provides a powerful tool for the discovery of natural antioxidants in Chinese medicines.

Ferroptosis inhibition protects vascular endothelial cells and maintains integrity of the blood-spinal cord barrier after spinal cord injury.

Maintaining the integrity of the blood-spinal cord barrier is critical for the recovery of spinal cord injury. Ferroptosis contributes to the pathogenesis of spinal cord injury. We hypothesized that ferroptosis is involved in disruption of the blood-spinal cord barrier. In this study, we administered the ferroptosis inhibitor liproxstatin-1 intraperitoneally after contusive spinal cord injury in rats. Liproxstatin-1 improved locomotor recovery and somatosensory evoked potential electrophysiological performance after spinal cord injury. Liproxstatin-1 maintained blood-spinal cord barrier integrity by upregulation of the expression of tight junction protein. Liproxstatin-1 inhibited ferroptosis of endothelial cell after spinal cord injury, as shown by the immunofluorescence of an endothelial cell marker (rat endothelium cell antigen-1, RECA-1) and ferroptosis markers Acyl-CoA synthetase long-chain family member 4 and 15-lipoxygenase. Liproxstatin-1 reduced brain endothelial cell ferroptosis in vitro by upregulating glutathione peroxidase 4 and downregulating Acyl-CoA synthetase long-chain family member 4 and 15-lipoxygenase. Furthermore, inflammatory cell recruitment and astrogliosis were mitigated after liproxstatin-1 treatment. In summary, liproxstatin-1 improved spinal cord injury recovery by inhibiting ferroptosis in endothelial cells and maintaining blood-spinal cord barrier integrity.

ROS-responsive exogenous functional mitochondria can rescue neural cells post-ischemic stroke.

Background: The transfer of mitochondria from healthy mesenchymal stem cells (MSCs) to injured MSCs has been shown to have potential therapeutic benefits for neural cell post-ischemic stroke. Specifically, functional mitochondria can perform their normal functions after being internalized by stressed cells, leading to host cell survival. However, while this approach shows promise, there is still a lack of understanding regarding which neural cells can internalize functional mitochondria and the regulatory mechanisms involved. To address this gap, we investigated the ability of different neural cells to internalize exogenous functional mitochondria extracted from MSCs. Methods: Functional mitochondria (F-Mito) isolated from umbilical cord derived-MSCs (UCMSCs) were labeled with lentivirus of HBLV-mito-dsred-Null-PURO vector. The ability of stressed cells to internalize F-Mito was analyzed using a mouse (C57BL/6 J) middle cerebral artery occlusion (MCAO) model and an oxygen-glucose deprivation/reoxygenation (OGD/R) cell model. The cell viability was measured by CCK-8 kit. Time-course of intracellular ROS levels in stressed cells were analyzed by DCFH-DA staining after OGD/R and F-Mito treatment. MitoSOX, Mitotracker and WGA labeling were used to assess the relationship between ROS levels and the uptake of F-Mito at the single-cell level. Pharmacological modulation of ROS was performed using acetylcysteine (ROS inhibitor). Results: Our findings demonstrate that neurons and endothelial cells are more effective at internalizing mitochondria than astrocytes, both in vitro and in vivo, using an ischemia-reperfusion model. Additionally, internalized F-Mito decreases host cell reactive oxygen species (ROS) levels and rescues survival. Importantly, we found that the ROS response in stressed cells after ischemia is a crucial determinant in positively mediating the internalization of F-Mito by host cells, and inhibiting the generation of ROS chemicals in host cells may decrease the internalization of F-Mito. These results offer insight into how exogenous mitochondria rescue neural cells via ROS response in an ischemic stroke model. Overall, our study provides solid evidence for the translational application of MSC-derived mitochondria as a promising treatment for ischemic stroke.