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Abstract Recognizing online handwriting Yi text is crucial for recording and preserving Yi literature. However, the scarcity of online handwriting Yi text datasets has limited relevant research, impeding the process of Yi informatization. In this work, we use synthetic data to train models, and an Online Handwriting Yi Text Recognition Network (YTRN) is proposed, which extracts robust character features to address the gap between synthetic and real data. YTRN adeptly learns the spatial structure features from path signature feature maps and captures trajectory features from the pen-tip trajectories. Subsequently, an innovative adaptive feature fusion module integrates these two sets of features to yield more comprehensive and robust character representations. Experiments on our newly collected Yi-OLHWDB2.0 dataset demonstrate that our method outperforms previous approaches, achieving an impressive 95.67% accuracy. This highlights the model’s effectiveness in extracting comprehensive and robust features from path signature maps and pen-tip trajectories, significantly enhancing recognition accuracy and generalization.

Objective: This study aimed to explore the clinical effects of music therapy (MT) on premature infants in neonatal intensive care units (NICUs). Methods: A total of 78 premature infants in NICUs admitted from January 2021 to January 2022 in Wuhan Children’s Hospital, Tongji Medical College, and Huazhong University of Science and Technology were selected as the reference group and received routine management. Wuhan Children’s Hospital implemented MT from February 2022 to February 2023, and 74 premature infants in NICUs admitted during the same period were selected as the observation group. The corresponding management mode was implemented on the second day of NICU admission for premature infants. Amplitude-integrated electroencephalogram (aEEG) and neonatal behavioral neurological assessment (NBNA) were adopted to evaluate the brain function, specifically the neurological function of neonates, after the end of management. The degree of parent–child attachment was measured using the pictorial representation of attachment measure (PRAM) in a nonverbal (visual) manner. The temperature, pulse, respiratory rate (RR), the number and duration of crying, and incidence of complications were all compared between the two groups. Results: SPSS showed that no difference existed in the aEEG and NBNA scores between the two groups (P > 0.05). The distance of PRAM self-baby-distance was smaller in the observation group compared with the reference group (P < 0.05). Furthermore, there was no significant difference in temperature between the two groups (P > 0.05). The observation group had significantly lower pulse and RR values than the reference group (P < 0.05). Finally, no significant difference existed in the incidence of complications between the two groups (P > 0.05). Conclusion: MT has a certain application value for premature infants in NICUs and can thus be applied to newborns in other NICUs. However, further studies are required to completely verify the research results.

Abstract The Dongba manuscripts are a unique primitive pictographic writing system that originated among the Naxi people of Lijiang, China, boasting over a thousand years of history. The uniqueness of the Dongba manuscripts stems from their pronounced pictorial and ideographic characteristics. However, the digital preservation and inheritance of Dongba manu manuscripts face multiple challenges, including extracting its rich semantic information, recognizing individual characters, retrieving Dongba manuscripts, and automatically interpreting the meanings of Dongba manuscripts. Developing efficient Dongba character detection technology has become a key research focus, wherein establishing a standardized Dongba detection dataset is crucial for training and evaluating techniques. In this study, we have created a comprehensive Dongba manuscripts detection dataset covering various commonly used Dongba characters and vocabularies. Additionally, we propose a model named STEF. Firstly, the Swin Transformer extracts the complex structures and diverse shapes of Dongba manuscripts’ features. Then, by introducing a Feature Pyramid Enhancement Module, features of different sizes are cascaded to preserve multi-scale information. Subsequently, all features are fused in a FUSION module, resulting in features of various Dongba manuscript styles. Each pixel’s binarisation threshold is dynamically adjusted through a differentiable binarisation operation, accurately distinguishing between foreground Dongba manuscripts and background. Lastly, deformable convolution is introduced, allowing the model to dynamically adjust the convolution kernel’s size and shape based on the Dongba manuscripts’ size, thereby better capturing the detailed information of Dongba characters of different sizes. Experimental results show that STEF achieves a recall rate of 88.88%, a precision rate of 88.65%, and an F-measure of 88.76%, outperforming other text detection algorithms. Visualization experiments demonstrate that STEF performs well in detecting Dongba manuscripts of various sizes, shapes, and styles, especially in blurred handwriting and complex backgrounds.

Abstract Background Colorectal cancer (CRC) commonly exhibits tolerance to cisplatin treatment, but the underlying mechanisms remain unclear. Within the tumor microenvironment, macrophages play a role in resisting the cytotoxic effects of chemotherapy by engaging in efferocytosis to clear apoptotic cells induced by chemotherapeutic agents. The involvement of extracellular vesicles (EVs), an intercellular communicator within the tumor microenvironment, in regulating the efferocytosis for the promotion of drug resistance has not been thoroughly investigated. Methods We constructed GFP fluorescent-expressing CRC cell lines (including GFP-CT26 and GFP-MC38) to detect macrophage efferocytosis through flow cytometric analysis. We isolated and purified CRC-secreted EVs using a multi-step ultracentrifugation method and identified them through electron microscopy and nanoflow cytometry. Proteomic analysis was conducted to identify the protein molecules carried by CRC-EVs. MFGE8 knockout CRC cell lines were constructed using CRISPR-Cas9, and their effects were validated through in vitro and in vivo experiments using Western blotting, immunofluorescence, and flow cytometric analysis, confirming that these EVs activate the macrophage αvβ3-Src-FAK-STAT3 signaling pathway, thereby promoting efferocytosis. Results In this study, we found that CRC-derived EVs (CRC-EVs) enhanced macrophage efferocytosis of cisplatin-induced apoptotic CRC cells. Analysis of The Cancer Genome Atlas (TCGA) database revealed a high expression of the efferocytosis-associated gene MFGE8 in CRC patients, suggesting a poorer prognosis. Additionally, mass spectrometry-based proteomic analysis identified a high abundance of MFGE8 protein in CRC-EVs. Utilizing CRISPR-Cas9 gene edition system, we generated MFGE8-knockout CRC cells, demonstrating that their EVs fail to upregulate macrophage efferocytosis in vitro and in vivo. Furthermore, we demonstrated that MFGE8 in CRC-EVs stimulated macrophage efferocytosis by increasing the expression of αvβ3 on the cell surface, thereby activating the intracellular Src-FAK-STAT3 signaling pathway. Conclusions Therefore, this study highlighted a mechanism in CRC-EVs carrying MFGE8 activated the macrophage efferocytosis. This activation promoted the clearance of cisplatin-induced apoptotic CRC cells, contributing to CRC resistance against cisplatin. These findings provide novel insights into the potential synergistic application of chemotherapy drugs, EVs inhibitors, and efferocytosis antagonists for CRC treatment.

L-theanine (LTA) is a key component in tea plants and is commonly used as a functional ingredient and dietary supplement. In a recent study, we investigated the potential antiaging properties of LTA using a D-galactose-induced L6 skeletal muscle aging cell model. The results showed that LTA treatment improved cell viability and antioxidant enzyme activities while reducing levels of proinflammatory factors and advanced glycation end products. Additionally, LTA demonstrated the ability to decrease senescence-associated beta-galactosidase-positive cells, lower reactive oxygen species and MDA levels, preserve mitochondrial function, inhibit apoptosis, and alleviate cell cycle arrest in the G2/M phase. These protective effects may be attributed to the activation of the p53/p21/CDK4 pathway, regulation of cell cycle-related protein expression, promotion of cell proliferation, and regulation of apoptosis-related protein expression through the p53/Bax/Bcl-2 pathway.

Reinforced concrete structures play a pivotal role in island and reef engineering projects. Given the resource constraints typical of island regions, substituting traditional manufactured sand aggregate with reef limestone not only reduces reliance on river sand but also addresses the issue of disposing of waste reef limestone slag generated during excavation. However, the performance characteristics of reef limestone concrete, particularly its bond strength with reinforcing steel, warrant further investigation. This is particularly true for the bond–slip behavior of the reinforcement. This study aims to elucidate the effects of various parameters on the bond performance between steel and reef limestone concrete through central pullout tests. These parameters include the type and diameter of the reinforcement, bond length, and loading rate. The investigation encompasses the analysis of load–slip curves, bond failure modes, and variations in bond stress. Additionally, using the Abaqus software, a numerical simulation was conducted to analyze the mesoscopic stress characteristics, thereby revealing the mechanisms of bond formation and failure modes between steel reinforcement and reef limestone concrete. The results indicate that the bond–slip curve for reef limestone concrete reinforced with ribbed rebars and Glass Fiber-Reinforced Polymer (GFRP) rebars can be broadly categorized into four phases: minor slip, slip, decline, and residual, with the residual phase exhibiting a wave-like pattern. The predominant failure modes in reef limestone concrete are either pulling out or splitting. The bond stress in reef limestone concrete decreases with an increase in rebar diameter and bond length; conversely, it increases with the loading rate, although the ultimate slip decreases. The mesoscopic failure characteristics of reinforced reef limestone concrete, as simulated in Abaqus, are consistent with the experimental outcomes.

The global burden of liver disease is enormous, which highlights the need for effective hepatoprotective agents. It was reported that allicin exhibits protective effects against a range of diseases. In this study, we further evaluated allicin’s effect and mechanism in acute hepatic injury. Liver injury in mice was induced by intraperitoneal injection with 1% CCl4 (10 mL/kg/day). When the first dose was given, CCl4 was given immediately after administration of different doses of allicin (40, 20, and 10 mg/kg/day) as well as compound glycyrrhizin (CGI, 80 mg/kg/day), and then different doses of allicin (40, 20, and 10 mg/kg/day) as well as compound glycyrrhizin (CGI, 80 mg/kg/day) were administrated every 12 h. The animals were dissected 24 h after the first administration. The findings demonstrated a significant inhibition of CCl4-induced acute liver injury following allicin treatment. This inhibition was evidenced by notable reductions in serum levels of transaminases, specifically aspartate transaminase, along with mitigated histological damage to the liver. In this protective process, allicin plays the role of reducing the amounts or the expression levels of proinflammatory cytokines, IL-1β, IL-6. Furthermore, allicin recovered the activities of the antioxidant enzyme catalase (CAT) and reduced the production of malondialdehyde (MDA) in a dose-dependent manner, and also reduced liver Caspase 3, Caspase 8, and BAX to inhibit liver cell apoptosis. Further analysis showed that the administration of allicin inhibited the increased protein levels of Nuclear factor-erythroid 2-related factor 2 (Nrf2) and NAD(P)H:quinone oxidoreductase 1 (NQO1), which is related to inflammation and oxidative stress. The in vitro study of the LPS-induced RAW264.7 inflammatory cell model confirmed that allicin can inhibit important inflammation-related factors and alleviate inflammation. This research firstly clarified that allicin has a significant protective effect on CCl4-induced liver injury via inhibiting the inflammatory response and hepatocyte apoptosis, alleviating oxidative stress associated with the progress of liver damage, highlighting the potential of allicin as a hepatoprotective agent.

The mitogen-activated protein kinase (MAPK) cascades act as crucial signaling modules that regulate plant growth and development, response to biotic/abiotic stresses, and plant immunity. MAP3Ks can be activated through MAP4K phosphorylation in non-plant systems, but this has not been reported in plants to date. Here, we identified a total of 234 putative TaMAPK family members in wheat (Triticum aestivum L.). They included 48 MAPKs, 17 MAP2Ks, 144 MAP3Ks, and 25 MAP4Ks. We conducted systematic analyses of the evolution, domain conservation, interaction networks, and expression profiles of these TaMAPK–TaMAP4K (representing TaMAPK, TaMAP2K, TaMAP3K, and TaMAP4K) kinase family members. The 234 TaMAPK–TaMAP4Ks are distributed on 21 chromosomes and one unknown linkage group (Un). Notably, 25 of these TaMAP4K family members possessed the conserved motifs of MAP4K genes, including glycine-rich motif, invariant lysine (K) motif, HRD motif, DFG motif, and signature motif. TaMAPK3 and 6, and TaMAP4K10/24 were shown to be strongly expressed not only throughout the growth and development stages but also in response to drought or heat stress. The bioinformatics analyses and qRT-PCR results suggested that wheat may activate the MAP4K10–MEKK7–MAP2K11–MAPK6 pathway to increase drought resistance in wheat, and the MAP4K10–MAP3K8–MAP2K1/11-MAPK3 pathway may be involved in plant growth. In general, our work identified members of the MAPK–MAP4K cascade in wheat and profiled their potential roles during their response to abiotic stresses and plant growth based on their expression pattern. The characterized cascades might be good candidates for future crop improvement and molecular breeding.

Abstract Background RING (Really Interesting New Gene) zinc finger (RING-zf) proteins belong to an important subclass of zinc fingers superfamily, which play versatile roles during various developmental stages and in abiotic stress responses. Based on the conserved cysteine and histidine residues, the RING-zf domains are classified into RING-HC (C3HC4), RING-H2 (C3H2C3), RING-v, RING-D, RING-S/T, RING-G, and RING-C2. However, little is known about the function of the RING-zfs of wheat. Results In this study, 129 (93.5%) of 138 members were found in nucleus, indicating TaRING-zf were primarily engaged in the degradation of transcription factors and other nuclear-localized proteins. 138 TaRING-zf domains can be divided into four canonical or modified types (RING-H2, RING-HC, RING-D, and RING-M). The RING-M was newly identified in T. aestivum, and might represent the intermediate other states between RING-zf domain and other modified domains. The consensus sequence of the RING-M domain can be described as M-X2-R-X14-Cys-X1-H-X2-Cys-X2-Cys-X10-Cys-X2-Cys. Further interspecies collinearity analyses showed that TaRING-zfs were more closely related to the genes in Poaceae. According to the public transcriptome data, most of the TaRING-zfs were expressed at different 15 stages of plant growth, development, and some of them exhibited specific responses to drought/heat stress. Moreover, 4 RING-HC (TraesCS2A02G526800.1, TraesCS4A02G290600.1, TraesCS4B02G023600.1 and TraesCS4D02G021200.1) and 2 RING-H2 (TraesCS3A02G288900.1 and TraesCS4A02G174600.1) were significantly expressed at different development stages and under drought stress. These findings provide valuable reference data for further study of their physiological functions in wheat varieties. Conclusions Taken together, the characterization and classifications of the TaRING-zf family were extensively studied and some new features about it were revealed. This study could provide some valuable targets for further studies on their functions in growth and development, and abiotic stress responses in wheat.

Dark tea (containing Eurotium cristatum) and black tea have hypoglycemic effects. The black brick tea with fungal growth is obtained from black tea by adding E. cristatum, followed by steaming, pressing, fungal growth, and drying. However, the hypoglycemic effects of black brick tea are still unexplored. Here, we used black brick tea with fungal growth and black tea as raw materials to study their hypoglycemic effects in a hyperglycemic mice model. Both these types of black tea could lower the content of blood glucose and increase the content of hepatic glycogen by upregulating the proteins and mRNA expression of phosphatidylinositol-4,5-bisphosphate3-kinase, glycogen synthase, protein kinase B and phosphoinositide-dependent protein kinase-1 and downregulating the protein and mRNA expression of glycogen synthase kinase 3β. These regulatory steps were followed by high activities of glutathione peroxidase and superoxide dismutase and low content of malondialdehyde. These teas can promote blood glucose transport and absorption by upregulating the protein and mRNA expression of insulin receptor substrate-1 and glucose transporter-2 and stimulate glycolysis by upregulating the protein and mRNA expression of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2. Our study suggests that the black brick tea was more effective than the black tea in terms of hypoglycemic.

The ubiquitin-specific protease (UBP) family represents a type of deubiquitinase (DUB) that plays a significant role in plant abiotic stress responses, growth, and development. Although UBP genes have been characterized in many species, a genome-wide systematic analysis and the potential functions under abiotic stresses have not yet been reported in rice (Oryza sativa L.). In this study, we identified 21 UBP gene family members in the genome of rice, all of which had UCH domains containing short but highly conserved Cys-box and His-box. The DUF4220 and DUF594 domains, which are found in monocotyledonous plants, were unique. Phylogenetic analysis suggested that OsUBP7 and OsUBP12 genes had the closest genetic relationships with AtUBP12 and AtUBP13, implying that OsUBP7 and OsUBP12 might have similar biological functions to AtUBP12 and AtUBP13. Synteny analysis revealed that OsUBP genes were expanded through segmental duplication under purifying selection. Meanwhile, they had closer evolutionary relationships with monocotyledonous plants. Additionally, all OsUBP genes had an ABRE cis-element, which was related to abscisic acid (ABA) stress hormone response. Protein interaction analysis suggested that OsUBP22 might modulate gene regulation through OsGCN5- and OsADA2-mediated chromatin modification, and OsUBP10 might regulate salt stress through modulating OsDNA2_4 stability. Specifically, the expression profiles and the transcriptional levels of 11 representative OsUBPs suggested the potential important functions of OsUBP9 and OsUBP17 under drought and cold stresses. Taken together, our study provided a systematic analysis of OsUBPs and preliminarily explored their potential roles under abiotic stresses. Our results lay a foundation for further research on the functions of the OsUBP gene family under abiotic stresses.

As an improved method of the lattice Boltzmann method (LBM), the regularized lattice Boltzmann method (RLBM) has been widely used to simulate fluid flow. For solving high Reynolds number problems, large eddy simulation (LES) and RLBM can be combined. The computation of fluid flow problems often requires a large number of computational grids and large-scale parallel clusters. Therefore, the high scalability parallel algorithm of RLBM with LES on a large-scale cluster has been proposed in this paper. The proposed parallel algorithm can solve complex flow problems with large-scale Cartesian grids and high Reynolds numbers. In order to achieve computational load balancing, the domain decomposition method (DDM) has been used in large-scale mesh generation. Three mesh generation strategies are adopted, namely 1D, 2D and 3D. Then, the buffer on the grid interface is introduced and the corresponding 1D, 2D and 3D parallel data exchange strategies are proposed. For the 3D lid-driven cavity flow and incompressible flow around a sphere under a high Reynolds number, the given parallel algorithm is analyzed in detail. Experimental results show that the proposed parallel algorithm has a high scalability and accuracy on hundreds of thousands of cores.

Based on the high-fat diet of successfully modeled obese and hypercholesterolemic rats, using simvastatin as a positive control，the results of epigallocatechin gallate (EGCG, 83.3 mg·kg−1·d−1) + L-Theanine (16.7 mg·kg−1·d−1), EGCG (100 mg·kg−1·d−1) and L-theanine (100 mg·kg−1·d−1) continuously fed by gavage to high-fat diet SPF-grade SD rats for 56 d were compared to investigate the synergistic moderating effects of L-theanine and EGCG for the prevention of obesity and hypercholesterolemia. The results showed that compared with the normal group, there were significant differences (P

ObjectivesUnexplained recurrent spontaneous abortion (URSA) remains an intractable reproductive dilemma due to the lack of understanding of the pathogenesis. This study aimed to evaluate the preclinical evidence for the mesenchymal stromal cell (MSC) treatment for URSA.MethodsA meticulous literature search was independently performed by two authors across the Cochrane Library, EMBASE, and PubMed databases from inception to April 9, 2023. Each study incorporated was assessed using the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE) risk of bias tool. The amalgamated standardized mean difference (SMD) accompanied by 95% confidence interval (CI) were deduced through a fixed-effects or random-effects model analysis.ResultsA total of ten studies incorporating 140 mice were subjected to data analysis. The MSC treatment yielded a significant reduction in the abortion rate within the URSA model (OR = 0.23, 95%CI [0.17, 0.3], P0.05).ConclusionsThe findings underscore the considerable potential of MSCs in URSA therapy. Nonetheless, the demand for enhanced transparency in research design and direct comparisons between various MSC sources and administration routes in URSA is paramount to engendering robust evidence that could pave the way for successful clinical translation.

The present trial was performed to reveal the regulatory effects of L-theanine on the levels of lipopolysaccharide (LPS) endotoxin within different biofluids, as well as relevant inflammatory responses of dairy cattle under heat stress conditions. Thirty lactating Chinese Holstein dairy cattle (189 ± 47 d in milk, and 2 ± 1 parities) were allocated in a completely randomized design to each of 3 dietary treatments: the control (CON, 0 g/d per cow L-theanine), the low L-theanine dosage treatment (LL, 16 g/d per cow L-theanine), and the high L-theanine dosage treatment (HL, 32 g/d per cow L-theanine). This trial consisted of 38 d (7 d for adaption and 31 d for data and sample collection), and sample collection for rumen liquid, blood plasma or serum, and milk were conducted on the d 27 and 38, respectively. Dairy cattle were constantly exposed to environmental heat stress during this experiment according to the recorded temperature-humidity index (THI). In the LL treatment, LPS concentration in rumen liquid was higher (P

Abstract Background Emerging evidences have revealed that long non-coding RNAs (lncRNAs) have played critical roles in tumor occurrence and progression. LINC00641 has been reported to be involved in the initiation and development of several cancers in the recent years. However, the detailed biological role of LINC00641 in renal cell carcinoma (RCC) remains largely unclear. Methods In this study, the expression and biological function of LINC00641 were assessed in renal carcinoma both in vitro and in vivo. Cell proliferation, migration and colony formation assay were performed to explore the effect of LINC00641on growth, progression and invasion of RCC cell. qRT-PCR, flow cytometry and luciferase reporter assay and in vivo tumorigenicity assay were also carried out. Results The expression of LINC00641 was overexpressed in RCC tissues and cell lines, and high LINC00641 expression was correlated with tumor-node-metastasis stage. Furthermore, Silencing of LINC00641 remarkably inhibited the ability of cell proliferation, colony formation, and invasive capacities, as well as increasing the apoptotic rates of RCC cells in vitro. Mechanistically, miR-340-5p was validated to be targeted by LINC00641 and knockdown of miR-340-5p counteracted LINC00641 silencing-mediated inhibition of RCC progression. In addition, in vivo experiment confirmed the findings discovered in vitro. Conclusions These results suggested that LINC00641 promoted the progression of RCC by sponging miR-340-5p.

Sugars and phytohormones co-regulate various common responses and development processes in plants. The previous studies reveal that the cross-talk between sugar and phytohormone signaling affects cellular elongation in root and hypocotyl. The elongation of hypocotyl is vital for the initial growth and development of seedlings during the transit from skotomorphogenesis to photomorphogenesis. Here, we found that LecRK-Ⅷ.2 gene regulated hypocotyl elongation involving in cross-talk between sugar and BR signaling. The results showed that sugar treatment could partly recover the defect of brassinosteroid (BR) response in bak1 mutant. LecRK-Ⅷ.2 gene regualted hypocotyl elongation by integrating the signaling cross-talk among BR, sucrose, glucose and fructose. Expression analysis of BR signaling related genes in response to exogenous sugar and BR suggested that LecRK-Ⅷ.2 may play essential roles in regulating hypocotyl growth. We also studied the roles of sugar transporter genes (STP1 and STP13) during hypocotyl elongation in the lecrk-Ⅷ.2 mutants in response to sugar/BR. The results showed that STP1 and STP13 expression in lecrk-Ⅷ.2-1 mutants were higher than that of Col-0 plant under sugar-free conditions. The expressions of STP1 in lecrk-Ⅷ.2 mutants were down-regulated compared to Col-0 under glucose treatments. The expressions of STP13 in lecrk-Ⅷ mutants were up-regulated compared to Col-0 under sucrose treatments. Altogether, our work indicated that LecRK-Ⅷ.2 regulated hypocotyl elongation by mediating the signaling cross-talk among sucose, glucose and BR.

Graph neural networks (GNNs) have garnered significant attention for their ability to effectively process graph-related data. Most existing methods assume that the input graph is noise-free; however, this assumption is frequently violated in real-world scenarios, resulting in impaired graph representations. To address this issue, we start from the essence of graph structure learning, considering edge discovery and removal, reweighting of existing edges, and differentiability of the graph structure. We introduce virtual nodes and consider connections with virtual nodes to generate optimized graph structures, and subsequently utilize Gumbel-Softmax to reweight edges and achieve differentiability of the Graph Structure Learning (VN-GSL for abbreviation). We conducted a thorough evaluation of our method on a range of benchmark datasets under both clean and adversarial circumstances. The results of our experiments demonstrate that our approach exhibits superiority in terms of both performance and efficiency. Our implementation will be made available to the public.

In this paper, stabilization control of a class of delayed inertial neural networks (INN) is investigated. Employing matrix measure method and two Halanay-type inequalities, some succinct stabilization criteria in terms of algebraic inequalities are derived for the INN with time-varying delays under periodically intermittent control (PIMC) strategy. Moreover, more precise results are obtained to stabilize the INN with time-invariant delays by using comparison principle. Specifically, the criteria of matrix measure form proposed in this paper can be converted into LMI-type condition for the case of 2-norm, which provides a bridge between the matrix-measure method and the Lyapunov function method. Finally, two numerical examples validate the efficacy of the derived results. The comparative research shows that the proposed methods generalize and develop some known results.

Abstract DNA barcoding is currently an effective and widely used tool that enables rapid and accurate identification of plant species. The Orchidaceae is the second largest family of flowering plants, with more than 700 genera and 20,000 species distributed nearly worldwide. The accurate identification of Orchids not only contributes to the safe utilization of these plants, but also it is essential to the protection and utilization of germplasm resources. In this study, the DNA barcoding of 4 chloroplast genes (matK, rbcL, ndhF and ycf1) were used to provide theoretical basis for species identification, germplasm conservation and innovative utilization of orchids. By comparing the nucleotide replacement saturation of the single or combined sequences among the 4 genes, we found that these sequences reached a saturation state and were suitable for phylogenetic relationship analysis. The phylogenetic analyses based on genetic distance indicated that ndhF and ycf1 sequences were competent to identification at genus and species level of orchids in a single gene. In the combined sequences, matK + ycf1 and ndhF + ycf1 were qualified for identification at the genera and species levels, suggesting the potential roles of ndhF, ycf1, matK + ycf1 and ndhF + ycf1 as candidate barcodes for orchids. Based on the SNP sites, candidate genes were used to obtain the specific barcode of orchid plant species and generated the corresponding DNA QR code ID card that could be immediately recognized by electronic devices. This study provides innovative research methods for efficient species identification of orchids. The standardized and accurate barcode information of Orchids is provided for researchers. It lays the foundation for the conservation, evaluation, innovative utilization and protection of Orchidaceae germplasm resources.

The CCHC-type zinc finger proteins (CCHC-ZFPs) play versatile roles in plant growth, development and adaptation to the environment. However, little is known about functions of CCHC-ZFP gene family memebers in Triticum aestivum. In the present study, we identified a total of 50 TaCCHC-ZFP genes from the 21 wheat chromosomes, which were phylogenetically classified into eight groups based on their specific motifs and gene structures. The 43 segmentally duplicated TaCCHC-ZFP genes were retrieved, which formed 36 segmental duplication gene pairs. The collinearity analyses among wheat and other eight mono/dicots revealed that no gene pairs were found between wheat and the three dicots. The promoter analyses of the TaCCHC-ZFP genes showed that 636 environmental stress-responsive and phytohormone-responsive cis-elements. The gene ontology enrichment analysis indicated that all the TaCCHC-ZFP genes were annotated under nucleic acid binding and metal ion binding. A total of 91 MicroRNA (miRNA) binding sites were identified in 34 TaCCHC-ZFP genes according to the miRNA target analysis. Based on the public transcriptome data, the 38 TaCCHC-ZFP genes were identified as differentially expressed gene. The expression profiles of 15 TaCCHC-ZFP genes were verified by the quantitative real-time PCR assays, and the results showed that these genes were responsive to drought or heat treatments. Our work systematically investigated the gene structures, evolutionary features, and potential functions of TaCCHC-ZFP genes. It lays a foundation for further research and application of TaCCHC-ZFP genes in genetic improvement of T. aestivum.

Epigallocatechin-3-gallate (EGCG) and L-theanine (LTA), which are active ingredients in green tea, can inhibit fat accumulation; however, EGCG bioavailability is

The intestine is a key organ for the absorption of amino acids. L-theanine (LTA) is a structural analog of glutamine and a characteristic non-protein amino acid found in tea (Camellia sinensis) that regulates lipid and protein metabolism. The present study explored the role of LTA in intestinal amino acid absorption, protein synthesis, and its mechanisms. Overall, our findings suggest that LTA supplementation not only affects serum alkaline phosphatase (AKP), total protein (TP), and urea nitrogen (BUN) levels, but it also upregulates the mRNA and protein expression of amino acid transporters (EAAT3, EAAT1, 4F2hc, y+LAT1, CAT1, ASCT2, and B0AT1), and activates the mTOR signaling pathway. The downstream S6 and S6K1 proteins are regulated, and the expression of amino acid transporters is regulated. These findings suggest that LTA increases intestinal AA absorption, promotes protein metabolism, and increases nitrogen utilization by upregulating AAT expression, activating the mTOR signaling pathway, and phosphorylating the mTOR downstream proteins S6 and S6K1.

ObjectiveThis study aimed to explore the role of ACSL4 in CD8+ T cell tumor infiltration and outcomes of bladder cancer (BLCA) patients after immunotherapy.MethodsThe correlation between ACSL4 expression and tumor infiltration of immune cells was analyzed using the Tumor Immune Estimation Resource database. The prognostic significance of ACSL4 in BLCA was analyzed using Kaplan–Meier curves. Immunohistochemistry was used to detect CD8+ T cell infiltration in tumors with high and low ACSL4 expression obtained from patients at the Fudan University Shanghai Cancer Center. The relationships between immune checkpoint genes and immune response were analyzed using The Cancer Genome Atlas and IMvigor 210 cohorts. The molecular functions, cellular components, and biological processes involving ACSL4 were explored using Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment pathway analyses.ResultsThe expression level of ACSL4 was significantly correlated with the infiltration of CD8+ T cells in BLCA tumors (r = 0.192, P = 2.22e-04). Elevated ACSL4 was associated with suppressed tumor progression and better outcomes for BLCA patients. The higher expression level of ACSL4 predicted better immunotherapeutic responses and was associated with higher expression levels of core immune checkpoint genes, including CD274, CTLA4, PDCD1, and LAG3, compared with the low ACSL4 expression group.ConclusionThis study demonstrated for the first time that elevated ACSL4 correlated significantly with CD8+ T cell infiltration and contributed to better immunotherapeutic responses in BLCA patients. Furthermore, ACSL4 serves as a novel biomarker for predicting patient outcomes after immunotherapeutic treatments, which may improve the development of individualized immunotherapy for BLCA.

With a direct bandgap, two-dimensional (2D) ZnSe is a promising semiconductor material in photoelectric device fields. In this work, based on first-principles methods, we theoretically studied the modulation of the Schottky barrier height (SBH) by applying horizontal and vertical strains on graphene/ZnSe heterojunction. The results show that the inherent electronic properties of graphene and ZnSe monolayers are both well-conserved because of the weak van der Waals (vdW) forces between two sublayers. Under horizontal strain condition, the n(p)-type SBH decreases from 0.56 (1.62) eV to 0.21 (0.78) eV. By changing the interlayer distance in the range of 2.8 Å to 4.4 Å, the n(p)-type SBH decreases (increases) from 0.88 (0.98) eV to 0.21 (1.76) eV. These findings prove the SBH of the heterojunction to be tuned effectively, which is of great significance to optoelectronic devices, especially in graphene/ZnSe-based nano-electronic and optoelectronic devices.

Nicotine hydrochloride (NCT) has a good control effect on hemiptera pests, but its poor interfacial behavior on the hydrophobic leaf leads to few practical applications. In this study, a vesicle solution by the eco-friendly surfactant, sodium diisooctyl succinate sulfonate (AOT), was prepared as the pesticide carrier for NCT. The physical chemical properties of NCT-loaded AOT vesicles (NCT/AOT) were investigated by techniques such as dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), and cryogenic transmission electron microscopy (cryo-TEM). The results showed that the pesticide loading and encapsulation efficiency of NCT/AOT were 10.6% and 94.8%, respectively. The size of NCT/AOT vesicle was about 177 nm. SAXS and surface tension results indicated that the structure of the NCT/AOT vesicle still existed with low surface tension even after being diluted 200 times. The contact angle of NCT/AOT was always below 30°, which means it could wet the surface of the cabbage leaf well. Consequently, NCT/AOT vesicles could effectively reduce the bounce of pesticide droplets. In vitro release experiments showed that NCT/AOT vesicles had sustained release properties; 60% of NCT in NCT/AOT released after 24 h, and 80% after 48 h. Insecticidal activity assays against aphids revealed that AOT vesicles exhibited insecticidal activity and could have a synergistic insecticidal effect with NCT after the loading of NCT. Thus, the NCT/AOT vesicles significantly improved the insecticidal efficiency of NCT, which has potential application in agricultural production activities.

l-Theanine (LTA), a non-protein-derived amino acid, is widely used as a functional ingredient. Here, we established an immune-stressed mouse model by infecting mice with enterotoxigenic Escherichia coli (ETEC) to study the immunomodulatory effects of LTA with respect to nucleotide-binding oligomerization domain (NOD)-like receptor signaling in vivo. Compared to ETEC control group, LTA alleviated ileal tissue lesions, significantly attenuated interleukin-1 beta (IL-1β) and tumor necrosis alpha (TNFα) overexpression, which was consistent with their protein levels. Additionally, LTA inhibited the ETEC-induced overexpression of NOD2 and receptor-interacting protein 2 (RIP2) proteins and mRNAs, TGE-beta-activated kinase 1 (TAK1) and proteasome subunit alpha type-7 (PSMA7) mRNAs, and partially decreased nuclear factor-κB (NF-κB) p65 protein levels. Furthermore, it suppressed c-Jun NH2-terminal kinase 1/2 (JNK1/2), extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 mitogen-activated protein kinase (p38) phosphorylation. Thus, we demonstrated that LTA regulates the innate immunity of ETEC-induced immune-stressed mice via NOD1/2-NF-κB and NOD1/2-MAPK pathways.

EMT allows a polarized epithelium to lose epithelial integrity and acquire mesenchymal characteristics. Previously, we found that overexpression of the intracellular domain of Notch3 (N3ICD) can inhibit EMT in breast cancer cells. In this study, we aimed to elucidate the influence of N3ICD or N3ICD combined with the transmembrane domain (TD+N3ICD) on the expression and distribution of TJs/AJs and polar molecules. We found that although N3ICD can upregulate the expression levels of the above-mentioned molecules, TD+N3ICD can inhibit EMT more effectively than N3ICD alone. TD+N3ICD overexpression upregulated the expression of endogenous full-length Notch3 and contributed to correcting the position of TJs/AJs molecules and better acinar structures formation. Co-immunoprecipitation results showed that the upregulated endogenous full-length Notch3 could physically interact with E-ca in MDA-MB-231/pCMV-(TD+N3ICD) cells. Collectively, our data indicate that overexpression of TD+N3ICD can effectively inhibit EMT, resulting in better positioning of TJs/AJs molecules and cell-cell adhesion in breast cancer cells. Abbreviations: EMT: Epithelial-mesenchymal transition; TJs: Tight junctions; AJs: Adherens junctions; aPKC: Atypical protein kinase C; Crb: Crumbs; Lgl: Lethal (2) giant larvae; LLGL2: lethal giant larvae homolog 2; PAR: Partitioning defective; PATJ: Pals1-associated TJ protein

The effects of nanoparticles (NPs) on microbiota homeostasis and their physiological relevance are still unclear. Herein, we compared the modulation and consequent pharmacological effects of oral administration of (−)-epigallocatechin-3-gallate (EGCG)-loaded β-cyclodextrin (β-CD) NPs (EGCG@β-CD NPs) and EGCG on gut microbiota. EGCG@β-CD NPs were prepared using self-assembly and their influence on the intestinal microbiome structure was analyzed using a metagenomics approach. The “Encapsulation efficiency (EE), particle size, polydispersity index (PDI), zeta potential” of EGCG@β-CD NPs were recorded as 98.27 ± 0.36%, 124.6 nm, 0.313 and –24.3 mV, respectively. Surface morphology of EGCG@β-CD NPs was observed as spherical. Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and molecular docking studies confirmed that EGCG could be well encapsulated in β-CD and formed as EGCG@β-CD NPs. After being continuously administered EGCG@β-CD NPs for 8 weeks, the serum cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and liver malondialdehyde (MDA) levels in the rats were significantly decreased, while the levels of catalase (CAT) and apolipoprotein-A1 (apo-A1) in the liver increased significantly in the hyperlipidemia model of rats, when compared to the high-fat-diet group. Furthermore, metagenomic analysis revealed that the ratio of Verrucomicrobia/Bacteroidetes was altered and Bacteroidetes decreased in the high-fat diet +200 mg/kg·bw EGCG@β-CD NPs group, while the abundance of Verrucomicrobia was significantly increased, especially Akkermansia muciniphila in rat feces. EGCG@β-CD NPs could be a promising EGCG delivery strategy to modulate the gut microbiota, enhancing its employment in the prevention of hyperlipidemia.

l-theanine, a non-protein-derived amino acid, is widely used as a functional ingredient in foods. Here, we evaluated the in vivo effects of l-theanine on glutamine metabolism in healthy rats and in a stressed rat model, which was established by infecting rats with enterotoxigenic Escherichia coli (E44813). The results showed that l-theanine significantly enhanced the synthesis of glutamine and stabilized the intestinal tract in healthy rats. In E44813-stressed rats, l-theanine could replace l-glutamine, thereby alleviating intestinal stress by reducing the expression of inflammatory factors, enhancing the activities of antioxidant enzymes, and regulating glutamine metabolism by modulating its synthesis. Treatment with a combination of l-theanine and l-glutamine was superior to treatment with l-glutamine alone in enhancing the activity of antioxidant enzymes, reducing the expression of inflammatory factors, and increasing the intestinal villi height and crypt depth.

The aim of this study was to investigate the oxidative and proliferative effects of grape seed extract (GSE). Piglet intestinal epithelial cells (IPEC1) were selected as an unstressed cell model, or they were exposed to 400 μM H2O2 to establish a H2O2-stimulated cell model. The glutathione (GSH) and total antioxidant capacity in response to GSE addition were tested in the unstressed and H2O2-stimulated cell models. The relative mRNA levels of antioxidant or antioxidant enzymes and apoptosis-related genes were measured by Real-Time RT-PCR. In the unstressed status, the cell survival ratio and GSH increased with the addition of GSE at 1 and 10 μg/mL but diminished at 60 μg GSE/mL. The addition of 1 μg/mL GSE upregulated the mRNA expression levels of B-Cell Lymphoma protein-2 (Bcl-2), cysteine aspartases-3 (Caspase-3), cysteine aspartases-8 (Caspase-8) and glutathione peroxidase-1(GPx-1), while it downregulated that of Bcl2-associated X protein (Bax), copper-zinc superoxide dismutase, glutathione S-transferase (GST), thioredoxin, thioltransferase and thioredoxin reductase. As GSE reached 60 μg/mL, the tumor protein p53 (p53) and caspase-8 gene expressions were upregulated. In stressed status, 1 and 10 μg GSE/mL promoted the increase of GSH. H2O2- induced increases in Bax, p53, and Caspase-3 mRNA expressions were attenuated by the subsequent addition of 1 μg GSE/mL and promoted the gene expression of tumor necrosis factor-α, GPx-1 and thioltransferase (Ttas). Treatment with 60 μg GSE/ml resulted in a significant reduction in Bax, p53, manganese superoxide dismutase and GST mRNA expressions. These results indicate that GSE exhibits antioxidative and proliferative functions on unstressed IPEC1 cells at low and medium levels and oxidative and antiproliferative functions at high levels.

Epigallocatechin gallate (EGCG) and L-theanine (LTA) are important bioactive components in tea that have shown promising effects on nutrient metabolism. However, whether EGCG alone or combined with LTA can regulate the glucose, lipid, and protein metabolism of healthy rats remains unclear. Therefore, we treated healthy rats with EGCG or the combination of EGCG and LTA (EGCG+LTA) to investigate the effects of EGCG on nutrient metabolism and the role of LTA in the metabolism-regulatory effects of EGCG. The results showed that compared with the control group, EGCG activated insulin and AMP-activated protein kinase (AMPK) signals, thus regulating glucose, lipid, and protein metabolism. Compared with EGCG, EGCG+LTA enhanced hepatic and muscle glycogen levels and suppressed phosphorylation of AMPK, glycogen synthase 2, mammalian target of rapamycin, and ribosomal protein S6 kinase. In addition, EGCG+LTA inhibited the expression of liver kinase B1, insulin receptor and insulin receptor substrate, and promoted the phosphorylation level of acetyl-CoA carboxylase. Furthermore, both EGCG and EGCG+LTA were harmless for young rats. In conclusion, EGCG activated AMPK and insulin pathways, thereby promoting glycolysis, glycogen, and protein synthesis and inhibiting fatty acid (FA) and cholesterol synthesis. However, LTA cooperated with EGCG to promote glycogen metabolism and suppressed the effect EGCG on FA and protein synthesis via AMPK signals.

l-Theanine (LTA), a non-protein-derived amino acid, is isolated from tea and is used as dietary supplement or beverage ingredient. This study established an enterotoxigenic Escherichia coli (ETEC)-infected mouse model of oxidative stress to examine the antioxidative mechanisms of LTA in vivo. LTA regulated non-enzymatic antioxidative activity and increased catalase (CAT) mRNA expression in non-ETEC-infected mice. In addition, LTA decreased oxidative parameters and the activity and mRNA levels of CAT in ETEC-infected mice. LTA increased superoxide dismutase (SOD) activity and mRNA expression of CuZn-SOD, ECSOD and glutathione peroxidase 1 (Gpx1); it restored MnSOD mRNA levels as well. Our results demonstrate that LTA has antioxidative properties via regulation of non-enzymatic pathways. In addition, LTA protects against superoxide and hydrogen peroxide via SOD or Gpx1 as the first and second defence line. Thus, LTA differentially regulates antioxidative enzymes depending on the dose and pathophysiological conditions through transcription, post-transcriptional and post-translational mechanisms in mice.

The safety, efficacy and stability of natural antioxidants have been the focus of research in the food industry, with the aim of rapidly analyzing and controlling the quality of rosemary and its extracts, a novel analytical method involving high-performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous determination of rosmarinic acid, carnosol, carnosic acid, oleanolic acid and ursolic acid in rosemary. Chromatographic separation was conducted with gradient elution mode by using a Zorbax SB-C18 column (4.6 mm × 250 mm, 5 μm) with mobile phases of methanol and 0.6% acetic acid. The drift tube temperature of ELSD was 70 °C, and the pressure of nebulizer nitrogen gas was 40 Psi. The method developed has high sensitivity (with limits of detection from 1.3 to 8.6 μg/mL), acceptable linearity over the tested concentrations (with correlation coefficients from 0.991 to 0.999), good repeatability (with intra- and inter-day CV less than 3.1% for all analytes) and satisfactory accuracy (with recovery between 95.5% and 100.8%). The method has been demonstrated as a powerful tool for the functional ingredients analysis and quality control of rosemary and its extracts in a cost- and time-effective manner.

A new homozygous humanized transgenic mouse strain, HLA-A2.1(+/+)HLA-DP4(+/+) hCD4(+/+)mCD4(-/-)IAβ(-/-)β2m(-/-) (HLA-A2/DP4), was obtained by crossing the previously characterized HLA-A2(+/+)β2m(-/-) (A2) mouse and our previously created HLA-DP4(+/+) hCD4(+/+)mCD4(-/-)IAβ(-/-) (DP4) mouse. We confirmed that the transgenes (HLA-A2, HLA-DP4, hCD4) inherited from the parental A2 and DP4 mice are functional in the HLA-A2/DP4 mice. After immunizing HLA-A2/DP4 mice with a hepatitis B DNA vaccine, hepatitis B virus-specific antibodies, HLA-A2-restricted and HLA-DP4-restricted responses were observed to be similar to those in naturally infected humans. Therefore, the present study demonstrated that HLA-A2/DP4 transgenic mice can faithfully mimic human cellular responses. Furthermore, we reported four new HLA-DP4-restricted epitopes derived from HBsAg that were identified in both vaccinated HLA-A2/DP4 mice and HLA-DP4-positive human individuals. The HLA-A2/DP4 mouse model is a promising preclinical animal model carrying alleles present to more than a quarter of the human population. This model should facilitate the identification of novel HLA-A2- and HLA-DP4-restricted epitopes and vaccine development as well as the characterization of HLA-DP4-restricted responses against infection in humans.

The photocatalyst anatase titanium dioxide (TiO2) films were immobilized via sol-gel technique on foam nickel modified with SiO2 films as transition layer (indicated as TiO2/SiO2 films). The structural properties of TiO2/SiO2 composite films were characterized using TG-DSC, XRD, FE-SEM, and BET. The photocatalytic activities of immobilized TiO2/SiO2 films were tested through photocatalytic degradation reactions of gaseous acetaldehyde under ultraviolet (UV) light irradiation. The substrate modification with transition layer of SiO2 films improved the specific surface area (SSA) of substrate and the acetaldehyde adsorption on photocatalyst film enormously and resulted in great enhancement of photocatalytic activity and stability. The degradation ratio of gaseous acetaldehyde (concentration of 100 ppmv) on TiO2/SiO2 photocatalysts reached 100% after 100 minutes of irradiation. The photocatalytic activity of TiO2/SiO2 films reduced about 15% after 9 consecutive runs, which was more stable than that of TiO2 films, and was completely recovered after heating at 300∘C for 1 hour.