As an environmental pollutant, fluoride is widespread in the natural environment in different forms, and drinking water is the primary way of exposure in human and animals. Structural damage to the central nervous system may occur in human and animals after fluoride exposure, which can lead to cognitive dysfunction. However, the mechanism of cognitive impairment caused by fluoride remains unclear. In this research, a fluoride-exposed model of mice and HT-22 cells was established to explore the neurotoxic mechanisms of fluoride. In vitro, CCK-8 results showed that HT-22 cells decreased with the increase in fluoride concentrations, and the morphology appeared abnormal. Similarly, laser confocal microscopy revealed that the number of axons and dendrites decreased with the increase in fluoride concentrations. Western blot results showed that the expression level of synaptic and cytoskeleton-associated proteins decreased in fluoride groups. In vivo, the mice exhibited losses in body and brain weights in the fluoride groups compared with the control group. The step-down test demonstrated that the cognitive ability of mice in the fluoride groups significantly decreased compared with that in the control group. Western blot results showed that the expression of synaptic and cytoskeleton-associated proteins decreased in the low-fluoride group compared with the control group, and qRT-PCR results showed that PSD95 expression decreased significantly compared with the control group. These results indicated that cognitive impairment induced by fluoride is involved in the morphological damage of synapse and the abnormal expression of synaptic proteins.

Lead (Pb) is a neurotoxic environmental pollutant that causes cognitive dysfunction and metabolic disturbances in the brain. Glucose transporter 1 (GLUT1) is essential for cerebral glucose metabolism, and altered expression may impair glucose uptake and utilization, synaptic plasticity, and cognitive performance. We found that acute Pb exposure impaired learning, memory, and anxiety-related behavior of mice. Fasting blood glucose concentrations increased in Pb-exposed mice, whereas GLUT1 expression in the brain was significantly decreased. GLUT1 overexpression alleviated Pb-induced synaptic structural and functional damage in Neuro-2a cells, whereas GLUT1 knockdown exacerbated these effects. Co-treatment with Morinda officinalis oligosaccharides (MOOs) improved behavioral performance, restored GLUT1 expression, and reduced Pb-induced synaptic pathology, whereas BAY-876 aggravated neurological impairment. In conclusion, GLUT1 is a key regulator of Pb-induced neurotoxicity. These findings provide potential therapeutic strategies for preventing the neurotoxic effects of Pb.

Bisphenol A (BPA), an environmental contaminant that migrates from food packaging, poses significant risks to embryonic development. This study investigated the protective effects and mechanisms of vitamin C (l-ascorbic acid) against BPA-induced embryotoxicity using Xenopus laevis embryos and P19 stem cells under a simultaneous exposure design. A multiassay approach was applied, including developmental bioassays, histological analysis, single-cell gel electrophoresis, and molecular profiling. The results showed that vitamin C significantly alleviated BPA-induced mortality, malformations, growth retardation, neurodevelopmental impairments, and notochord damage. Mechanistically, vitamin C restored the expression of antioxidant genes (gpx4, sod, ppard, and cat.2), reduced reactive oxygen species generation and oxidative DNA damage, and suppressed apoptosis via modulation of the ATM/Chk2/p53 signaling pathway. These findings demonstrate that vitamin C mitigates BPA-induced embryotoxicity by limiting oxidative stress and DNA damage-mediated apoptosis under coexposure conditions, supporting further evaluation of antioxidant-based protection across different exposure regimens and in combination with other antioxidants.

Lead (Pb) is a heavy metal environmental pollutant with strong biological toxicity. Our previous study suggested that Pb may impair learning and memory by disrupting cytoskeletal structure and inhibiting the expression of synaptic plasticity-related proteins in mice. However, the exact mechanism of Pb-induced cytoskeletal damage remains unclear. In this study, Neuro-2a cells and Kunming mice were used to explore the neurotoxic mechanism of Pb. The actin dynamics were observed via laser confocal microscopy. The ATP levels and ATPase activity in Neuro-2a cells was measured. In addition, the mRNA and protein expression levels of RhoA/ROCK/Cofilin signaling pathway in brain tissues and Neuro-2a cells was measured, and the mRNA expression levels of glucose metabolism rate-limiting enzymes were detected. Our results showed that Pb induces nerve cell damage and cytoskeletal abnormalities. Western blot and qRT-PCR analyses revealed that Pb activated the RhoA/ROCK/Cofilin signaling pathway. Additionally, ATPase activity significantly decreased following Pb treatment, whereas ATP levels markedly increased in the 50 μM Pb group. In addition, Pb disrupts brain glucose metabolism through affect the transcription of rate-limiting enzymes of glucose metabolism. Overall, these findings suggest that Pb activates the RhoA/ROCK/Cofilin signaling pathway, leading to cytoskeletal damage. Moreover, Pb exposure alters glucose metabolism enzyme activity and ATP production, disrupting the balance between F-actin and G-actin and ultimately affecting neuronal structure and function. These results may provide a better understanding of lead-induced nerve damage.

Enhanced glycolysis and elevated lactic acid (LA) production are observed during sudden death syndrome (SDS) in broilers. However, the mechanism underlying LA-induced cardiomyocyte damage and heart failure in fast-growing broilers remains unclear. In this study, chicken embryo cardiomyocytes (CECs) were cultured and treated with LA to investigate LA-induced CEC injury and its mechanism, aiming to develop strategies to prevent LA-induced SDS in broilers. Results showed that LA inhibited CEC proliferation and contraction whereas inducing apoptosis. Furthermore, LA disrupted mitochondrial ultrastructure, reduced mitochondrial membrane potential, activated mitophagy, and disturbed mitochondrial dynamics. Treatment with Mdivi-1, a selective Drp1 inhibitor, improved CEC viability, restored mitochondrial network integrity, reduced reactive oxygen species production, and inhibited LA-induced apoptosis. These findings suggest that LA-induced cardiomyocyte injury during SDS in broilers is associated with mitochondrial damage and increased mitochondrial fission. The inhibition of mitochondrial hyperfission by Mdivi-1 effectively preserves CEC morphology, structure, and function, playing a critical role in preventing LA-induced damage. This study provides a foundation for strategies to prevent and control SDS in broilers.

Previous studies have demonstrated that oxidative stress and melanogenesis are regulated by the Wnt/β-catenin signaling pathway. However, the precise mechanism by which the PEDF/Wnt/β-catenin axis modulates apoptosis and melanogenesis remains unclear. Cell viability and mortality rates were assessed using CCK-8 assays and lactate dehydrogenase (LDH) release assays. Mitochondrial ultrastructural changes were analyzed by transmission electron microscopy (TEM). Changes in the mitochondrial membrane potential (ΔΨm) were assessed using the JC-1 fluorescent probe. The effects of PEDF on protein and gene expression were evaluated by Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). As the concentration of H2O2 increased, the cell survival rate decreased, which activated the Wnt/β-catenin signaling pathway and increased the apoptosis rate and melanin production. PEDF reversed the H2O2-induced decrease in cell viability and increase in mortality rate in Melan-a cells while ameliorating the impairment of the mitochondrial membrane potential. PEDF ameliorated H2O2-induced protein damage and lipid peroxidation and reduced apoptosis in Melan-a cells. PEDF treatment significantly decreased the protein expression levels of β-catenin, Wnt3a, and Dvl2 (P < 0.05) and reduced the protein levels of Bax and Caspase-3 (P < 0.01) in H2O2- and BML-284-treated Melan-a cells. Furthermore, PEDF significantly reduced the H2O2- and BML-284-induced increases in the MITF and TYR protein levels (P < 0.01). These results suggest that PEDF can reduce H2O2-induced oxidative damage and melanin production in Melan-a cells by inhibiting the activation of Wnt/β-catenin signaling pathway activation. These findings provide a theoretical basis for human oxidative stress and pigment deposition-related diseases.

To address the issue of instability induced by direct-drive wind turbines connected to weak AC grids, a grid-connected asymptotic tracking stabilization control strategy for direct-drive wind turbines based on quantitative trajectory constraints is proposed. First, an equivalent mathematical model of the direct-drive wind turbine unit is established, with the dq-axis coupling caused by unmodeled dynamics, power fluctuations, and operational mode switching regarded as inherent disturbances of the system, thereby enhancing modeling accuracy. Secondly, a grid-connected asymptotic tracking stabilization control method for wind turbines based on quantitative trajectory constraints is proposed. This method ensures transient response trajectory constraints throughout the process while guaranteeing steady-state tracking ability and robust operation of the overall system, ensuring that the entire transient response trajectory remains within the specified constraint range, thereby improving the stable operation of the wind turbine grid-connected system. Finally, a simulation model is built in PSCAD for comparative validation. The simulation results demonstrate that the proposed control strategy enhances the dynamic performance of the full transient response trajectory, effectively improving the stable operation capability of the overall system.

Florfenicol (FLO), a widely used antibiotic, can cause early embryonic death and impaired angiogenesis. However, the mechanism by which FLO inhibits angiogenesis and its role in FLO-induced embryonic toxicity are not yet fully understood. This study aimed to investigate the effect of FLO on the proliferation and migration of mouse bEnd.3 endothelial cells (bECs) and the underlying mechanism, given the crucial role of endothelial cells in angiogenesis. The results showed that FLO treatment at 7.5-15 μg/mL significantly inhibited proliferation and migration in a time-dependent manner, with stronger effects observed after 48 h compared to 24 h. RNA sequencing analysis identified 1351 genes differentially expressed in response to FLO. Functional analysis indicated that FLO disrupted ATP metabolism, angiogenesis, vasculature development, and actin filament organization. The F-actin cytoskeleton remodeling was morphologically confirmed, and activation of the ROCK/Cofilin signaling pathway was detected. Subsequent experiments demonstrated that pharmacological inhibition of the pathway normalized cytoskeletal rearrangement and promoted angiogenesis, as evidenced by enhanced wound closure and increased cell viability. Our results demonstrated that ROCK-mediated F-actin remodeling plays a crucial role in the FLO-induced inhibition of proliferation and migration in bECs. These findings may explain why FLO inhibited angiogenesis. However, further investigations are necessary to validate our findings in in vivo experiments.

Candida albicans is the most leading cause of life-threatening fungal invasive infections, especially for vulvovaginal candidiasis (VVC). Resistance and tolerance to common fungicide has risen great demands on alternative strategies for treating C. albicans infections. In the present study, ferroptosis has been proven to occur in C. albicans by directly exposed to FeSO4 via induing hallmarks of ferroptosis, including Fe2+ overload burden, ROS eruption and lipid peroxidation. Transcriptomic profile gave the great hints of the possible mechanism for fungal ferroptosis that FeSO4 disturb pathways associated to ribosome, tyrosine metabolism, triglyceride metabolism and thiamine metabolism, thus mobilizing death-related gene synthesis. Inspired by the results, a FeSO4-loaded hydrogel was prepared as an antifungal agent to treat C. albicans infection. This hydrogel exhibited excellent dressing properties and maintained superior antifungal activity by characterization tests. Besides, mice treated by this composite hydrogel displayed excellent therapeutic efficacy. These results highlighted the potential therapeutic use of FeSO4 as an innovative strategy in treating C. albicans infections by targeting ferroptosis.

Objective Coat color is an important characteristic and economic trait in domestic sheep. In this study, we explored the potential mechanisms and the signaling pathways involved in coat color regulation for sheep. Methods Isobaric tags for relative and absolute quantification (iTRAQ) technology was used to catalog global protein expression profiles in skin of sheep with black versus white coat color. Immunofluorescence was used to observe the expression localization of differential protein. Western blot and quantitative real time polymerase chain reaction (qRT-PCR) were used to evaluate their role in the coat color formation of sheep. Results A total of 136 differential proteins were obtained in different coat colors, including 101 up-regulated and 35 down-regulated. Pigmentation function entries were enriched through gene ontology annotation. Tyrosine metabolism and platelet activation signaling pathway were extracted by Kyoto encyclopedia of genes and genomes analysis. Apolipoprotein A-1 (APOA1) and fibrinogen alpha chain (FGA) were found to be critical differential proteins by the interaction of differential proteins in the direct-interaction network diagram. Strikingly, twenty candidate differential proteins were screened, from which beta-actin (ACTB) protein showed higher expression in white sheep skin, while albumin (ALB), APOA1, MAOA (amine oxidase) and FGA proteins showed higher expression in black sheep skin, which was validated by immunofluorescence, western blot, and qRT-PCR. Conclusion This study identified several novel proteins that may be involved in the coat color formation of sheep. The white and black sheep skin proteome profiles obtained provide a valuable resource for future research to understand the network of protein expression controlling skin physiology and melanogenesis in sheep.

DC converter stations have a high voltage level, a long transmission distance, and complex internal equipment, and contain power electronic devices, which seriously endanger the stable operation of the system itself and the active distribution network at the receiving end when faults occur. Accurate fault analysis and diagnosis are critical to the safe and stable operation of power systems. Traditional fault diagnosis methods often rely on a single source of information, leading to issues such as insufficient information utilization and incomplete diagnostic scope when applied to DC transmission systems. To address these problems, a fault diagnosis method for converter stations based on preliminary identification of the fault range and the fusion of evidence information of the switch signal and electrical quantity is proposed. First, the preprocessing of converter station sequential event recording (SER) events and a statistical analysis of event characteristics are completed to initially determine the range of the fault.Then, a fuzzy Petri net model and a BP neural network model are constructed on the basis of the fault data from a real-time digital simulation system (RTDS), and the corresponding evidence information of the switch signal and electrical quantity are obtained via iterative inference and deep learning methods. Finally, on the basis of D-S evidence theory, a comprehensive diagnosis result is obtained by fusing the switch and electric evidence information. Taking the fault data of a DC converter station as an example, the proposed method is analyzed and compared with the traditional method, which is based on single information. The results show that the proposed method can reliably and accurately identify fault points in the protected area of the converter station.

犬肥大细胞瘤(Canine mast cell tumor,MCT)是起源于真皮组织肥大细胞的恶性肿瘤,最常见于躯干、四肢和头颈部.犬肥大细胞瘤的临床表现多样,其表面可能脱毛、溃疡、红斑、色素过度沉着.一例患犬通过临床症状检查、血常规、血液生化、C-反应蛋白(C-reactive protein,CRP)、X射线以及病理组织学观察,最终确诊为犬的多发性肥大细胞瘤.对该犬实施了肿瘤及周围健康组织切除手术,并建议术后进行化疗,为后续提高犬多发性肥大细胞瘤治疗和诊断水平提供参考.

Fluoride compounds are abundant and widely distributed in the environment at various concentrations, which can seriously injure the human body. In this study, we aim to evaluate the effects of excessive fluoride exposure on the liver, kidney, and heart tissues of healthy female Xenopus laevis by administering NaF (0, 100, and 200 mg/L) in drinking water for 90 days. The expression level of procaspase-8, cleaved-caspase-8, and procaspase-3 proteins were determined by Western blot. Compared with the control group, the group exposed to NaF exhibited expression levels of procaspase-8, cleaved-caspase-8, and procaspase-3 proteins that were considerably upregulated at a concentration of 200 mg/L in the liver and kidney. The cleaved-caspase-8 protein expression in the group exposed to a high concentration of NaF was lower than that in the control group in heart. Histopathological results by hematoxylin and eosin staining showed that excessive NaF exposure caused necrosis of hepatocytes and vacuolization degeneration. Granular degeneration and necrosis in renal tubular epithelial cells were also observed. Moreover, hypertrophy of myocardial cells, atrophy of myocardial fibers and disorder of myocardial fibers were detected. These results demonstrated that NaF-induced apoptosis and the mediated death receptor pathway activation ultimately damaged the liver and kidney tissues. This finding offers a fresh perspective on the effects of F-induced apoptosis in X. laevis.

Florfenicol (FLO) has been shown to elicit diverse toxic effects in plants, insects, and mammals. Previously, our investigations revealed that FLO induced abnormal cardiac development and early embryonic mortality in chicken embryos. However, the effect of FLO on mitochondrial responses in stem cells remains unclear. In this study, we show that FLO significantly diminishes proliferation viability and obstructs the directed differentiation of P19 stem cells (P19SCs) into cardiomyocytes. Proteomic analysis revealed 148 differentially expressed proteins in response to FLO. Functional analysis has pinpointed FLO interference with biological processes associated with oxidative phosphorylation within the mitochondria. In alignment with the results of proteomic analysis, we confirmed that FLO inhibits the expression of both nuclear DNA-encoded and mitochondrial DNA-encoded subunits of the electron transport chain. Subsequent experiments demonstrated that FLO disrupts mitochondrial dynamics and induces the mitochondrial unfolded protein response to maintain mitochondrial homeostasis. These findings collectively highlight the significance of mitochondrial dynamics and the mitochondrial unfolded protein response to mediate the decreased proliferation viability and directed differentiation potential in P19SCs treated with FLO. In conclusion, this study provides a comprehensive overview of mitochondrial responses to FLO-induced cytotoxicity and enhances our understandings of the molecular mechanisms underlying FLO-induced embryonic toxicity.

Excessive fluoride intake poses health risks to humans and animals. Many studies have indicated that fluoride exposure can damage the cytoskeleton and synapses, which has negative effects on the intellectual development of humans and animals. Our previous study suggested that the RhoA/ROCK signalling pathway is activated by NaF exposure in HT-22 cells and plays a vital role in cytoskeletal assembly and synaptogenesis. However, the mechanism underlying RhoA/ROCK-mediated cytoskeletal injury induced by fluoride remains unclear. In this study, Neuro-2A cells and ICR mice were used to investigate the effects of RhoA/ROCK activation inhibition on NaF-induced synaptic dysfunction and cognitive impairment. We detected the expression of GAP, RhoA, ROCK1/ 2, and (p)-MLC in vivo and in vitro model. The results showed that NaF exposure activated the RhoA/ROCK/MLC signalling pathway. We measured the effects of RhoA/ROCK inhibition on synaptic injury and intellectual impairment induced by NaF exposure. In vitro, Y-27632 suppressed activated RhoA/ROCK, attenuated morphological and ultrastructural damage, and decreased the survival rate and synapse-functional protein expression caused by NaF. In vivo, the results showed that the RhoA/ROCK/MLC pathway was inhibited by fasudil and improved pathological damage in the hippocampus, cognitive impairment, and decreased expression of neurofunctional proteins induced by NaF. Overall, these results suggest that fasudil and Y-27632 can reverse neurotoxicity caused by fluoride exposure. Furthermore, inhibition of RhoA/ROCK may be a future treatment for CNS injury, and more detailed studies on other neurodegenerative disease models are required to confirm its effectiveness.

Background: Fluoride, an environmental contaminant, is ubiquitously present in air, water, and soil. It usually enters the body through drinking water and may cause structural and functional disorders in the central nervous system in humans and animals. Fluoride exposure affects cytoskeleton and neural function, but the mechanism is not clear.Methods: The specific neurotoxic mechanism of fluoride was explored in HT-22 cells. Cellular proliferation and toxicity detection were investigated by CCK-8, CCK-F, and cytotoxicity detection kits. The development morphology of HT-22 cells was observed under a light microscope. Cell membrane permeability and neuro-transmitter content were determined using lactate dehydrogenase (LDH) and glutamate content determination kits, respectively. The ultrastructural changes were detected by transmission electron microscopy, and actin homeostasis was observed by laser confocal microscopy. ATP enzyme and ATP activity were determined using the ATP content kit and ultramicro-total ATP enzyme content kit, respectively. The expression levels of GLUT1 and 3 were assessed by Western Blot assays and qRT-PCR.Results: Our results showed that fluoride reduced the proliferation and survival rates of HT-22 cells. Cytomor-phology showed that dendritic spines became shorter, cellular bodies became rounder, and adhesion decreased gradually after fluoride exposure. LDH results showed that fluoride exposure increased the membrane perme-ability of HT-22 cells. Transmission electron microscopy results showed that fluoride caused cells to swell, microvilli content decreased, cellular membrane integrity was damaged, chromatin was sparse, mitochondria ridge gap became wide, and microfilament and microtubule density decreased. Western Blot and qRT-PCR an-alyses showed that RhoA/ROCK/LIMK/Cofilin signaling pathway was activated by fluoride. F-actin/G-actin fluorescence intensity ratio remarkably increased in 0.125 and 0.5 mM NaF, and the mRNA expression of MAP2 was significantly decreased. Further studies showed that GLUT3 significantly increased in all fluoride groups, while GLUT1 decreased (p < 0.05). ATP contents remarkably increased, and ATP enzyme activity substantially decreased after NaF treatment with the control. Conclusion: Fluoride activates the RhoA/ROCK/LIMK/Cofilin signaling pathway, impairs the ultrastructure, and depresses the connection of synapses in HT-22 cells. Moreover, fluoride exposure affects the expression of glucose transporters (GLUT1 and 3) and ATP synthesis. Sum up fluoride exposure disrupts actin homeostasis, ultimately affecting structure, and function in HT-22 cells. These findings support our previous hypothesis and provide a new perspective on the neurotoxic mechanism of fluorosis.

Bisphenol A (BPA) has been reported to injure the developing and adult brain. However, the underlying mechanism still remains elusive. This study used neuro-2a cells as a cellular model to investigate the neurotoxic effects of BPA. Microtubule-associated protein 2 (MAP2) and tau protein maintain microtubule normal function and promote the normal development of the nervous system. Synaptophysin (SYP) and drebrin (Dbn) proteins are involved in regulating synaptic plasticity. Cells were exposed to the minimum essential medium (MEM), 0.01% (v/v) DMSO, and 150 μM BPA for 12, 24, or 36 h. Morphological analysis revealed that the cells in the BPA-treated groups shrank and collapsed compared with those in the control groups. CCK-8 and lactate dehydrogenase assay (LDH) assays showed that the mortality of neuro-2a cells increased as the BPA treatment time was prolonged. Ultrastructural analysis further revealed that cells demonstrated nucleolar swelling, dissolution of nuclear and mitochondrial membranes, and partial mitochondrial condensation following exposure to BPA. BPA also decreased the relative protein expression levels of MAP2, tau, and Dbn. Interestingly, the relative protein expression levels of SYP increased. These results indicated that BPA inhibited the proliferation and disrupted cytoskeleton and synaptic integrity of neuro-2a cells.

雄性英国短毛猫近月余出现逆呕、咳喘等症状.通过实验室检查、内窥镜检查、质子泵抑制性实验等方法进行诊断治疗,内窥镜检查中见食管处病变特征及稀薄反流液.综合诊断与治疗结果,该猫初步诊断为反流性食管炎.通过质子泵抑制剂治疗后,预后良好.

Bisphenol A (2,2-bis(4′-hydroxyphenyl) propane, BPA) is a well-known endocrine-disrupting compound that is widely used in various daily products and exhibits embryonic development toxicity and genotoxicity. However, the affected signaling pathways involved in embryonic development especially the interactions of involved proteins remain unclear. In our previous study (Ge et al., 2021), BPA induces DNA damage and apoptosis in Xenopus embryos, resulting in multiple malformations of larvae. However, the signaling pathways induced for apoptosis response to DNA damage are still not well elucidated. Here, we systematically elucidated the enriched pathways affected by BPA and illustrated the interactions of involved proteins. Results indicated that BPA affected multiple embryonic development pathways including Hippo, TGF-β, Wnt, and Notch pathways. Furthermore, the protein-protein interaction network suggested that the c-Abl/YAPY357/p73 pathway may play a key role in apoptosis induction in response to DNA damage. P19 embryonal carcinoma stem cells, as a developmental toxicity model, were treated with different BPA concentrations to establish an in vitro model to verify the role of the c-Abl/YAPY357/p73 pathway in apoptosis. BPA triggered DNA damage and significantly upregulated the expression levels of c-Abl, phosphorylated YAPY357, phosphorylated p73Y99, and cleaved caspase-3 protein (p < 0.05), thus decreasing cell viability and transcriptionally activating the p73 target genes Bax and Puma. These data suggested that BPA activated the c-Abl/YAPY357/p73 pathway in response to DNA damage. Imatinib, an inhibitor of tyrosine kinase c-Abl, significantly downregulated the elevated expression levels of p-YAPY357, p-p73Y99 and cleaved caspase-3 (p < 0.05) caused by BPA and then ameliorated the cell index of P19 cells in the BPA-treated group. Therefore, this substance restrained the phosphokinase activity of c-Abl and suppressed the c-Abl/YAPY357/p73 pathway. Results showed that the c-Abl/YAPY357/p73 pathway served as a mechanism for caspase-3 activation that induced the apoptosis response to DNA damage stress.