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BACKGROUND: Neuroblastoma (NB) is the most prevalent and deadliest pediatric solid tumor. With of over 50% of high-risk neuroblastoma cases relapse, the imperative for novel drug targets and therapeutic strategies is accentuated. In neuroblastoma, the existence of tumor-associated macrophages (TAMs) correlates with an unfavorable patient prognosis. However, the clinical relevance and prognostic implications of regulatory genes linked to TAMs infiltration in neuroblastoma remain unclear, and further study is required. METHODS: We conducted a comprehensive analysis utilizing transcriptome expression profiles from three primary datasets associated with neuroblastoma (GSE45547, GSE49710, TARGET) to identify hub genes implicated in immune evasion within neuroblastoma. Subsequently, we utilized single-cell RNA sequencing analysis on 17 clinical neuroblastoma samples to investigate the expression and distribution of these hub genes, leading to the identification of TNFAIP3. The above three public databases were merged to allowed for the validation of TNFAIP3's molecular functions through GO and KEGG analysis. Furthermore, we assessed TNFAIP3's correlation with immune infiltration and its potential immunotherapeutic impact by multiple algorithms. Our single-cell transcriptome data revealed the role of TNFAIP3 in macrophage polarization. Finally, preliminary experimental verifications to confirm the biological functions of TNFAIP3-mediated TAMs in NB. RESULTS: A total of 6 genes related to immune evasion were screened and we found that TNFAIP3 exhibited notably higher expression in macrophages than other immune cell types, based on the scRNA-sequencing data. GO and KEGG analysis showed that low expression of TNFAIP3 significantly correlated with the activation of multiple oncogenic pathways as well as immune-related pathways. Then validation affirmed that individuals within the TNFAIP3 high-expression cohort could potentially derive greater advantages from immunotherapeutic interventions, alongside exhibiting heightened immune responsiveness. Deciphering the pseudotime trajectory of macrophages, we revealed the potential of TNFAIP3 in inducing the polarization of macrophages towards the M1 phenotype. Finally, we confirmed that patients in the TNFAIP3 high expression group might benefit more from immunotherapy or chemotherapy as substantiated by RT-qPCR and immunofluorescence examinations. Moreover, the role of TNFAIP3 in macrophage polarization was validated. Preliminary experiment showed that TNFAIP3-mediated TAMs inhibit the proliferation, migration and invasion capabilities of NB cells. CONCLUSIONS: Our results suggest that TNFAIP3 was first identified as a promising biomarker for immunotherapy and potential molecular target in NB. Besides, the presence of TNFAIP3 within TAMs may offer a novel therapeutic strategy for NB.
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Boletus aereus Fr. ex Bull. stands out as a delectable edible mushroom with high nutritional and medicinal values, featuring polysaccharides as its primary nutrient composition. In our continuous exploration of its beneficial substances, a novel polysaccharide (BAPN-1) with a molecular weight of 2279 kDa was prepared. It was identified as a glucan with a backbone composed of the residues â4)-α-Glcp-(1â and â4,6)-α-Glcp-(1â connected in a proportion of 5:1 and a ß-Glcp-(1â side residue attached at C6 of the â4,6)-α-Glcp-(1â residue. Biologically, BAPN-1 exhibited broad-spectrum antiproliferative activities against various NHL cells, including HuT-78, OCI-LY1, OCI-LY18, Jurkat, RL, and Karpas-299, with IC50 values of 0.73, 1.21, 3.18, 1.52, 3.34, and 4.25 mg/mL, respectively. Additionally, BAPN-1 significantly induced cell cycle arrest in the G2/M phase and caused apoptosis of NHL cells. Mechanistically, bulk RNA sequencing and Western blot analysis revealed that BAPN-1 could upregulate cyclin B1 and enhance cleaved caspase-9 expression through the inhibition of FGFR3 and RAF-MEK-ERK signaling pathways. This work supports the improved utilization of B. aereus in high-value health products.
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Antineoplásicos , Apoptose , Proliferação de Células , Linfoma não Hodgkin , Polissacarídeos , Humanos , Proliferação de Células/efeitos dos fármacos , Linhagem Celular Tumoral , Apoptose/efeitos dos fármacos , Antineoplásicos/farmacologia , Antineoplásicos/química , Linfoma não Hodgkin/tratamento farmacológico , Polissacarídeos/química , Polissacarídeos/farmacologia , Polissacarídeos/isolamento & purificação , Peso Molecular , Basidiomycota/químicaRESUMO
Due to the increasing demand for higher security and low-cost energy storage systems, the main research focus has been developing a suitable substitute for lithium-ion batteries. Aqueous zinc ion batteries (AZIBs) are considered the best alternative to lithium-ion batteries in large-scale energy storage devices. Owing to its high capacity, vanadate is a promising cathode material for AZIBs. The crystallographic orientation of cathode materials dramatically influences the rate performance and cycling life. Here, Mg0.57V5O12·2.3H2O (MgVO) with favorable (001) crystal orientation and significantly improved electrochemical performance is prepared by a simple stirring method. The crystal growth orientations of MgVO are altered by adjusting the aging time of the reactant solution. The (001)-orientated grain growth of MgVO delivers a 232.5 mA h g-1 capacity at 5 A g-1 with a 94% capacity retention rate after 1400 cycles. The zinc ion storage performance of MgVO demonstrates that the orientation-controlled method can design effective cathode materials for high-performance ZIBs.
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Emotions, bodily sensations and movement are integral parts of musical experiences. Yet, it remains unknown i) whether emotional connotations and structural features of music elicit discrete bodily sensations and ii) whether these sensations are culturally consistent. We addressed these questions in a cross-cultural study with Western (European and North American, n = 903) and East Asian (Chinese, n = 1035). We precented participants with silhouettes of human bodies and asked them to indicate the bodily regions whose activity they felt changing while listening to Western and Asian musical pieces with varying emotional and acoustic qualities. The resulting bodily sensation maps (BSMs) varied as a function of the emotional qualities of the songs, particularly in the limb, chest, and head regions. Music-induced emotions and corresponding BSMs were replicable across Western and East Asian subjects. The BSMs clustered similarly across cultures, and cluster structures were similar for BSMs and self-reports of emotional experience. The acoustic and structural features of music were consistently associated with the emotion ratings and music-induced bodily sensations across cultures. These results highlight the importance of subjective bodily experience in music-induced emotions and demonstrate consistent associations between musical features, music-induced emotions, and bodily sensations across distant cultures.
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Música , Humanos , Música/psicologia , Sensação , Comparação Transcultural , Acústica , Emoções , Percepção AuditivaRESUMO
Whole milk powder and skimmed milk powder are suitable for different groups of people due to their differences in composition. Therefore, a rapid classification method for whole milk powder and skimmed milk powder is urgently needed. In this paper, a novel strategy based on laser-induced breakdown spectroscopy (LIBS) and feature processing methods combined with logistic regression (LR) was constructed for the classification of milk powder. A LR classification model based on mini-batch gradient descent (MGD) was employed first. As indicated by the research results, the accuracy of the MGD-LR model for the milk powder samples in the test set is 96.33% and the modeling time is 33.07 s. The modeling efficiency is low and needs to be improved. Principal components analysis (PCA) and mutual information (MI) were used as feature processing methods to reduce the high dimensional LIBS data into fewer features for improving the modeling efficiency of the classification model. The research results indicate that the accuracy of the PCA-MGD-LR model and the MI-MGD-LR model for the test set of milk powder samples was 99.33% and 99.67%, respectively. Compared with MGD-LR model, the modeling efficiency of PCA-MGD-LR and MI-MGD-LR models has increased by 89.7% and 74.8%, respectively. The results fully demonstrate the feasibility of rapid milk powder classification based on LIBS and feature processing methods combined with LR, and it will provide a new technology for the identification and classification of milk powder.
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Lasers , Leite , Humanos , Animais , Leite/química , Pós/análise , Modelos Logísticos , Análise Espectral/métodosRESUMO
Knowledge about the impact of different dissolved oxygen (DO) on the composition and function of gut bacteria of aquatic insects is largely unknown. Herein, we constructed freshwater environments with different DOs (hypoxia: 2.50 ± 0.50, normoxia: 7.00 ± 0.50, and hyperoxia: 13.00 ± 0.50 mg/L) where aquatic firefly Aquatica leii larvae lived for three months. Their gut flora was analyzed using the combination of 16S rRNA amplicon sequencing and metagenomics. The results showed no difference in alpha diversity of the gut flora between A. leii inhabiting various DOs. However, the relative abundance of several bacterial lineages presented significant changes, such as Pseudomonas. In addition, bacterial genes with an altered relative abundance in response to various DOs were primarily related to metabolism. The alteration of these functions correlated with the DO change. This is the first to uncover structure of gut flora under various DOs in aquatic insect larvae.
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Milk powder can provide the necessary nutrients for the growth of infants, and the level of its energy value is an important factor in the measurement of its nutritional value. Therefore, the measurement of the energy value in milk powder is of great significance for the nutritional health of infants. In this study, samples of 32 different brands of milk powder were selected for spectral analysis, and laser-induced breakdown spectroscopy (LIBS) combined with deep belief network (DBN), back propagation (BP) neural network, and long short-term memory (LSTM) models was used to achieve quantitative analysis of the energy value of the milk powder. The experimental results revealed that the LSTM model outperformed the DBN and BP models in terms of accuracy, with a mean relative error (MREP) of 1.0029%, which was 73.03% lower than that of DBN (3.7186%) and 69.53% lower than that of BP (3.2914%). Moreover, the determination coefficient (RP2) value improved significantly from 0.9341 for DBN and 0.9766 for BP to 0.9984. In addition, the root mean square error (RMSEP) decreased to 0.2140 from 0.7042 for DBN and 0.9051 for BP. These results demonstrate that the LSTM model has superior predictive performance compared to the other models. Therefore, the combination of LIBS and LSTM can accurately measure the energy value of milk powder and provide an effective and feasible means for its commercial measurement.
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Memória de Curto Prazo , Leite , Lactente , Humanos , Animais , Pós , Memória de Longo Prazo , LasersRESUMO
Background: Total hip arthroplasty (THA) has emerged as a pivotal approach for addressing femoral neck fractures (FNFs), a prevalent type of fracture in older people. Restoring joint functionality following surgery significantly contributes to patients' overall well-being. Therefore, the implementation of rational and effective rehabilitation exercises is crucial. Objective: This study aims to investigate the impact of phased rehabilitation management on patients with FNFs who have undergone THA. Methods: In this prospective comparative study, a total of 89 patients were enrolled. Among them, 49 patients underwent phased rehabilitation management, while 40 patients received conventional postoperative rehabilitation. The evaluation encompassed a range of assessments, including the Harris Hip Score (HHS) for the evaluation of hip joint function, the Five-Times-Sit-to-Stand Test (FTSST) to quantify lower limb muscle strength, the Barthel Index to assess activities of daily living, and the Visual Analog Scale (VAS) to measure pain intensity. Furthermore, preoperative and postoperative serum levels of interleukin-6 (IL-6), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were diligently measured to provide a comprehensive understanding of the patient's inflammatory responses. Results: Overall, the study group exhibited a significantly higher average HHS after surgery compared to the control group. Notably, the VAS scores at 1 day and 3 days post-surgery in both groups demonstrated a reduced trend compared to preoperative values. This trend was more significant in the study group compared to the control group. The time taken for the FTSST in patients undergoing phased rehabilitation management was less compared to the control group. Furthermore, phased rehabilitation management was associated with more significant improvements in activities of daily living. Notably, IL-6 levels were higher in both the study and control groups at 1 day postoperatively than before surgery, while they decreased at 3 days postoperatively compared to the 1-day mark. The study group exhibited significantly lower levels of CRP (mg/L) and ESR (mm/h) compared to the control group. Conclusions: Implementing phased rehabilitation management for patients with FNFs following THA improves hip joint function, lower limb muscle strength, daily living activities, pain intensity, and inflammatory response.
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Artroplastia de Quadril , Fraturas do Colo Femoral , Humanos , Idoso , Artroplastia de Quadril/reabilitação , Atividades Cotidianas , Estudos Prospectivos , Interleucina-6 , Resultado do Tratamento , Fraturas do Colo Femoral/cirurgiaRESUMO
The mechanisms underlying the toxicity of environmental stress are unclear for marine macrobenthos. Copper/Cu has posed the most serious threats to amphioxus, an ancient and model benthic cephalochordate. Herein, a dynamic change in the physiological parameters (GR, SOD, ATP, and MDA) was detected with ROS accumulation in Branchiostoma belcheri exposed to 0.3 mg·L-1 Cu. Transcriptomes and microRNAomes of B. belcheri were generated to investigate the molecular mechanisms by which this amphioxus copes with Cu exposure. Time-specific genes identified at different time points after exposure were involved in the stimulus and immune response, detoxification and ionic homeostasis, aging and the nervous system, sequentially, with prolongation of exposure time, forming a dynamic process of molecular response to Cu stress. In total, 57 differentially expressed miRNAs were identified under Cu stress. Transcriptomics-miRNAomics analyses indicate that these miRNAs targeted genes associated with many key biological processes such as xenobiotics degradation, oxidative stress, and energy metabolism. The constructed miRNA-mRNA-pathway network uncovered a broad post-transcriptional regulatory mechanism in B. belcheri to cope with Cu stress. Overall, this integrated analyses show that enhanced defense response, accelerated ROS elimination, and repressed ATP production constitute a comprehensive strategy to cope with Cu toxicity in the ancient macrobenthos.
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Anfioxos , MicroRNAs , Animais , Transcriptoma , Cobre/toxicidade , Cobre/metabolismo , Espécies Reativas de Oxigênio/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Trifosfato de Adenosina/metabolismoRESUMO
Calcium is the main mineral responsible for healthy bone growth in infants. Laser-induced breakdown spectroscopy (LIBS) was combined with a variable importance-based long short-term memory (VI-LSTM) for the quantitative analysis of calcium in infant formula powder. First, the full spectra were used to establish PLS (partial least squares) and LSTM models. The R2 and root-mean-square error (RMSE) of the test set (R P2 and R M S E P) were 0.1460 and 0.0093 in the PLS method, respectively, and 0.1454 and 0.0091 in the LSTM model, respectively. To improve the quantitative performance, variable selection based on variable importance was introduced to evaluate the contribution of input variables. The variable importance-based PLS (VI-PLS) model had R P2 and R M S E P of 0.1454 and 0.0091, respectively, whereas the VI-LSTM model had R P2 and R M S E P of 0.9845 and 0.0037, respectively. Compared with the LSTM model, the number of input variables in the VI-LSTM model was reduced to 276, R P2 was improved by 114.63%, and R M S E P was reduced by 46.38%. The mean relative error of the VI-LSTM model was 3.33%. We confirm the predictive ability of the VI-LSTM model for the calcium element in infant formula powder. Thus, combining VI-LSTM modeling and LIBS has great potential for the quantitative elemental analysis of dairy products.
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Cálcio , Fórmulas Infantis , Lactente , Humanos , Cálcio/análise , Pós , Memória de Curto Prazo , Análise Espectral/métodos , LasersRESUMO
Photodeposition of dual-cocatalysts Pt-Cr or Rh-Cr on the surface of ZnIn2S4 is used to achieve overall water splitting. Compared with the hybrid loading of the Pt element and Cr element, the formation of the Rh-S bond results in space separation of the Rh element and Cr element. The Rh-S bond and space separation of cocatalysts promote the transfer of bulk carriers to the surface and inhibit self-corrosion.
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We report herein that TSPAN32 is a key node factor for Philadelphia (Ph+) leukemia pathogenesis. We found that TSPAN32 expression was repressed by BCR-ABL and ectopic TSPAN32 expression upon Imatinib treatment inhibited the proliferation of Ph+ cell lines. Tspan32 overexpression significantly prevented BCR-ABL induced leukemia progression in a murine model and impaired leukemia stem cell (LSC) proliferation. LSCs represent an obstacle for chronic myeloid leukemia (CML) elimination, which continually replenish leukemia cells and are associated with disease relapse. Therefore, the identification of essential targets that contribute to the survival and self-renewal of LSCs is important for novel curative CML. Mechanistically, TSPAN32 was shown to interact with PTEN, increased its protein level and caused a reduction in PI3K-AKT signaling activity. We also found that TSPAN32 was repressed by BCR-ABL via the suppression of an important transcription factor, TAL1. Ectopic expression of TAL1 significantly increased TSPAN32 mRNA and protein level, which indicated that BCR-ABL repressed TSPAN32 transcription by decreasing TAL1 expression. Overall, we identified a new signaling axis composed of "BCR-ABL-TAL1-TSPAN32-PTEN-PI3K-AKT". Our findings further complement the known mechanisms underlying the transformation potential of BCR-ABL in CML pathogenesis. This new signaling axis also provides a potential means to target PI3K-AKT for CML treatment.
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Leucemia Mielogênica Crônica BCR-ABL Positiva , PTEN Fosfo-Hidrolase , Tetraspaninas , Animais , Camundongos , Mesilato de Imatinib/farmacologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Tetraspaninas/metabolismoRESUMO
Chemical fertilizers are important for effectively improving soil fertility, promoting crop growth, and increasing grain yield. Therefore, methods that can quickly and accurately measure the amount of fertilizer in the soil should be developed. In this study, 20 groups of soil samples were analyzed using laser-induced breakdown spectroscopy, and partial least squares (PLS) and random forest (RF) models were established. The prediction performances of the models for the chemical fertilizer content and pH were analyzed as well. The experimental results showed that the R 2 and root mean square error (RMSE) of the chemical fertilizer content in the soil obtained using the full-spectrum PLS model were .7852 and 2.2700 respectively. The predicted R 2 for soil pH was .7290, and RMSE was .2364. At the same time, the full-spectrum RF model showed R 2 of .9471 (an increase of 21%) and RMSE of .3021 (a decrease of 87%) for fertilizer content. R 2 for the soil pH under the RF model was .9517 (an increase of 31%), whereas RMSE was .0298 (a decrease of 87%). Therefore, the RF model showed better prediction performance than the PLS model. The results of this study show that the combination of laser-induced breakdown spectroscopy with RF algorithm is a feasible method for rapid determination of soil fertilizer content.
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Purinostat Mesylate (PM) is a novel highly selective and active HDAC I/IIb inhibitor, and the injectable formulation of PM (PMF) based on the compound prescription containing cyclodextrin completely can overcome PM's poor solubility and improves its stability and pharmacokinetic properties. Here, we showed that PM effectively repressed the survival of Ph+ leukemia cells and CD34+ leukemia cells from CML patients in vitro. In vivo studies demonstrated that PMF significantly prevented BCR-ABL(T315I) induced CML progression by restraining leukemia stem cells (LSCs), which are insensitive to chemotherapy and responsible for CML relapse. Mechanism studies revealed that targeting HDAC I/IIb repressed several important factors for LSCs survival including c-Myc, ß-Catenin, E2f, Ezh2, Alox5, and mTOR, as well as interrupted some critical biologic processes. Additionally, PMF increased glutamate metabolism in LSCs by increasing GLS1. The combination of PMF and glutaminase inhibitor BPTES synergistically eradicated LSCs by altering multiple key proteins and signaling pathways which are critical for LSC survival and self-renewal. Overall, our findings represent a new therapeutic strategy for eliminating LSCs by targeting HDAC I/IIb and glutaminolysis, which potentially provides a guidance for PMF clinical trials in the future for TKI resistance CML patients.
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Mitochondria are the leading organelle for energy metabolism. The toxic effects of environmental toxicants on mitochondrial morphology, energy metabolism, and their determination of cell fate have already been broadly studied. However, minimal research exists on effects of environmental toxicants such as pesticides on mitochondrial energy metabolism at in vitro subcellular level, particularly from an omics perspectives (e.g., metabolomics). Here, German cockroach (Blattella germanica) was fed diets with (0.01 and 0.001 mg/mL) and without abamectin, and highly purified fat body mitochondria were isolated. Swelling measurement confirmed abnormal mitochondrial swelling caused by abamectin stress. The activity of two key mitochondrial energy metabolism-related enzymes, namely succinic dehydrogenase and isocitrate dehydrogenase, was significantly affected. The metabolomic responses of the isolated mitochondria to abamectin were analyzed via untargeted liquid chromatography/mass spectrometry metabolomics technology. Fifty-two differential metabolites (DMs) were identified in the mitochondria between the 0.001 mg/mL abamectin-fed and the control groups. Many of these DMs were significantly enriched in pathways involved in ATP production and energy consumption (e.g., oxidative phosphorylation, TCA cycle, and pentose phosphate pathway). Nineteen of the DMs were typically related to energy metabolism. This study is valuable for further understanding mitochondrial toxicology under environmental toxicants, particularly its subcellular level.
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Phenyllactic acid (PLA) has been demonstrated to possess antibacterial activity and capacity to prolong food shelf life. However, studies on the performance of PLA in inhibiting Staphylococcus aureus and its effectiveness when applied to dairy products are largely lacking. Here, antibacterial activity (planktonic and biofilm states) of PLA against S. aureus CICC10145 (S. aureus_45) were investigated. The results showed that PLA inhibited growth of S. aureus_45 and formation of S. aureus_45 biofilm. Next, the antibacterial action target of PLA was uncovered from both physiological and phenotypic perspectives. The results showed that PLA decreased cell metabolic activity and cell viability, damaged cell membrane integrity, triggered leakage of intracellular contents (DNA, proteins, and ATP), and caused oxidative stress damage and morphological deformation of S. aureus_45. In practical application, the antibacterial activity of PLA against S. aureus_45 cells was further confirmed in skim milk and cheese as dairy food models, and the antibacterial effects can be adequately maintained during storage for 21 d, at least at 4°C. These findings suggested that PLA could be a potential candidate for controlling S. aureus outgrowth in dairy foods.
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Queijo , Infecções Estafilocócicas , Animais , Staphylococcus aureus , Queijo/microbiologia , Leite/microbiologia , Infecções Estafilocócicas/veterinária , Antibacterianos/farmacologia , PoliésteresRESUMO
Heterostructured double-phase composites are promising electrode candidates for high-performance secondary metal batteries due to their superior capacity and ion transfer kinetics compared with the pristine phase. Herein, a Zn3V3O8/VO2 (ZVO/VO) heterostructure with abundant phase boundaries was designed as the cathode for aqueous zinc-ion batteries (ZIBs). The preparation method is based on a solid pre-intercalation approach, and the Zn content in the ZVO/VO heterostructure can be precisely controlled. The electrochemical performance of ZVO/VO containing different amounts of Zn, pristine ZVO, and VO phases was compared. ZVO/VO showed superior capacity and cycling stability compared to pristine ZVO and VO. The ZVO/VO heterostructure showed a capacity of 328.4 mA h g-1 at 0.3 A g-1 after 200 cycles. The long-term cycling performance of ZVO/VO was evaluated at 3 A g-1, and it delivered a capacity retention of 90.5% after 1000 cycles. The ion storage mechanism of the ZVO/VO electrode was analyzed by ex situ X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). This work provides a simple strategy for designing vanadium-based heterostructure composites as advanced cathodes for ZIBs.
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Aquatic insects are well-adapted to freshwater environments, but metabolic mechanisms of such adaptations, particularly to primary environmental factors (e.g., hypoxia, water pressure, dark light, and abundant microbes), are poorly known. Most firefly species (Coleoptera: Lampyridae) are terrestrial, but the larvae of a few species are aquatic. We generated 24 global metabolomic profiles of larvae and adults of Aquatica leii (freshwater) and Lychnuris praetexta (terrestrial) to identify freshwater adaptation-related metabolites (AARMs). We identified 110 differentially abundant metabolites (DAMs) in A. leii (adults vs. aquatic larvae) and 183 DAMs in L. praetexta (adults vs. terrestrial larvae). Furthermore, 100 DAMs specific to aquatic A. leii larvae were screened as AARMs via interspecific comparisons (A. leii vs. L. praetexta), which were primarily involved in antioxidant activity, immune response, energy production and metabolism, and chitin biosynthesis. They were assigned to six categories/superclasses (e.g., lipids and lipid-like molecules, organic acids and derivatives, and organoheterocyclic compound). Finally, ten metabolic pathways shared between KEGG terms specific to aquatic fireflies and enriched by AARMs were screened as aquatic adaptation-related pathways (AARPs). These AARPs were primarily involved in energy metabolism, xenobiotic biodegradation, protection of oxidative/immune damage, oxidative stress response, and sense function (e.g., glycine, serine and threonine metabolism, drug metabolism-cytochrome P450, and taste transduction), and certain aspects of morphology (e.g., steroid hormone biosynthesis). These results provide evidence suggesting that abundance changes in metabolomes contribute to freshwater adaptation of fireflies. The metabolites identified here may be vital targets for future work to determine the mechanism of freshwater adaptation in insects.