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Objective: To investigate the immune mechanism of osteosarcoma (OS)-specific markers to mitigate bone destruction in the aggressive OS, prone to recurrence and metastasis. Methods: Gene expression patterns from the Gene Expression Omnibus (GEO) database (GSE126209) were analyzed using weighted gene co-expression network analysis (WGCNA), protein-protein interaction (PPI) analysis, least absolute shrinkage and selection operator (LASSO) modeling, and survival analysis to identify charged multivesicular body protein 4C (CHMP4C). Subsequently, its role in regulating the immune system and immune cell infiltration was explored. CHMP4C expression and signaling molecules in OS were assessed in osteosarcoma cell lines (MG63, U2OS, HOS) and hFOB1.19 cells using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The impact of CHMP4C upregulation and interference on OS-related signaling molecules in MG63 cells was studied. Functional validation of CHMP4C in MG63 OS cells was confirmed through cell counting Kit-8 (CCK-8), transwell, and colony formation assays. In vivo experiments were conducted using Specific Pathogen Free (SPF)-grade male BALB/C nude mice for OS xenograft studies. Results: Based on the gene expression profiles analysis of six osteosarcoma samples and six normal tissue samples, we identified 1,511 upregulated DEGs and 5,678 downregulated DEGs in normal tissue samples. A significant positive correlation between the "yellow-green" module and OS was found through WGCNA analysis. Expression levels of CHMP4C, phosphorylated Glycogen Synthase Kinase 3ß (p-GSK3ß), and ß-catenin were notably higher in U2OS, HOS, and MG63 OS cells than in hFOB1.19 human osteoblasts. Overexpressing CHMP4C in MG63 OS cells upregulated CHMP4C, p-GSK3ß, and ß-catenin while downregulating GSK3ß, leading to increased proliferation and migration of MG63 cells. Conversely, interrupting CHMP4C had the opposite effect. High expression of CHMP4C significantly accelerated the growth of OS in nude mice, resulting in substantial upregulation of CHMP4C, p-GSK3ß, and ß-catenin expression and suppression of Glycogen Synthase Kinase 3ß (GSK3ß) expression in OS tissues. Conclusion: CHMP4C may serve as a specific immunomodulatory gene for OS. Its activation of the Wnt/ß-catenin signaling pathway, mainly by increasing the phosphorylation echelon of GSK3ß, promotes the invasion and spread of OS.
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The "schisandra-evodia" herb pair (S-E) is a herbal preparation to treat Alzheimer's disease (AD). This study aims to investigate the therapeutic efficacy and potential mechanism of S-E in AD rats, utilizing pharmacodynamic assessments and serum- and urine-based metabolomic analyses. Pharmacodynamic assessments included Morris water maze test, hematoxylin-eosin staining and immunohistochemistry experiments. The results of the study showed that the AD model was successful; the S-E significantly enhanced long-term memory and spatial learning in AD rats. Meanwhile, S-E notably ameliorated Aß25-35-induced cognitive impairment, improved hippocampal neuron morphology, decreased Aß deposition in the hippocampus and mitigated inflammatory damage. We then analyzed serum and urine samples using UPLC-MS/MS to identify potential biomarkers and metabolic pathways. Metabolomic analysis revealed alterations in 40 serum metabolites and 38 urine metabolites following S-E treatment, predominantly affecting pathways related to taurine and hypotaurine metabolism, linoleic acid metabolism, α-linolenic acid metabolism, glycerophospholipid metabolism and arachidonic acid metabolism. This study elucidates the biochemical mechanism underlying AD and the metabolic pathway influenced by S-E, laying the groundwork for future clinical applications.
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Doença de Alzheimer , Metaboloma , Metabolômica , Ratos Sprague-Dawley , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/metabolismo , Animais , Metabolômica/métodos , Ratos , Cromatografia Líquida de Alta Pressão/métodos , Masculino , Metaboloma/efeitos dos fármacos , Metaboloma/fisiologia , Espectrometria de Massas em Tandem/métodos , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/administração & dosagem , Biomarcadores/sangue , Biomarcadores/urina , Hipocampo/metabolismo , Hipocampo/efeitos dos fármacos , Peptídeos beta-Amiloides/metabolismoRESUMO
Schisandra chinensis and Evodia rutaecarpa are traditional Chinese herbs used to treat neurodegenerative diseases. This study investigates the combined effects of SC and ER on learning and memory in an Alzheimer's disease rat model and their underlying mechanisms. Methods: High-performance liquid chromatography was employed to analyze the primary active constituents of Schisandra and Evodia. The effects of the combined treatment of Schisandra and Evodia on learning and memory in an Alzheimer's disease rat model were evaluated through Morris water maze and Hematoxylin-Eosin staining experiments. Immunohistochemical analysis was conducted to investigate the impact of S-E on Aß1-42 and P-tau proteins. Western blotting and real-time quantitative polymerase chain reaction were utilized to quantify the expression of pivotal proteins and genes within the BDNF/TRKB/CREB and GSK-3ß/Tau pathways. Results: The treatment group exhibited significant neuroprotective effects, ameliorating learning and memory impairments in the Alzheimer's disease rat model. The treatment regimen modulated the activity of the BDNF/TRKB/CREB and GSK-3ß/Tau pathways by influencing the expression of relevant genes, thereby reducing the generation of Aß1-42 and P-Tau proteins and inhibiting the deposition of senile plaques. Furthermore, among the three treatment groups, the combined treatment demonstrated notably superior therapeutic effects on Alzheimer's disease compared to the single-drug treatment groups.
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The soluble crude polysaccharides from Dioscorea opposita (DOP1 and DOP2) were prepared and characterized. DOP1 and DOP2 obtained carbohydrate (65.71% and 70.18%, respectively), uronic acid (63.71% and 24.84%, respectively) and protein (8.09% and 9.51%, respectively) with molecular weight of 49.24 kDa and 21.62 kDa, respectively. DOP samples were mainly composed of mannose, glucose, galacturonic acid, galactose, and glucuronic acid. The digestibility in vitro, antioxidant activity and intestinal peristalsis effect were then investigated. DOP1 and DOP2 were degraded with decreased molecular weights (39.58 kDa and 18.56 kDa respectively), increased reducing sugar contents (from 16.95% to 19.27%; 12.45% to 15.50% respectively) and free monosaccharides (from 0.89% to 1.42%; 0.90% to 1.14% respectively) after gastric digestion. Both DOP1 and DOP2 were resistant to intestinal digestion, suggesting that DOP samples can be considered as a dietary fiber. Additionally, DOP1 and DOP2 exhibited antioxidant activities positively correlated with the concentration and remained the activities after gastrointestinal digestion in vitro. Furthermore, DOP reduced the fluorescence intensity significantly, indicating DOP can promote the intestinal peristalsis of zebrafish larvae (5 pdf) at 500 µg/mL. Therefore, DOP1 and DOP2 have a better functionality as dietary fibers, including antioxidant activity and intestinal peristalsis promotion, which can be developed as functional foods.
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Antioxidantes , Dioscorea , Animais , Antioxidantes/farmacologia , Peixe-Zebra/metabolismo , Dioscorea/metabolismo , Peristaltismo , Polissacarídeos/farmacologia , Polissacarídeos/metabolismo , Fibras na DietaRESUMO
The edible films composed ofcarboxymethyl cellulose (CMC), glycerol, mucilage from Chinese yam (DOM)and ZnO nanoparticles (ZnO-NPs), were prepared by a casting method. To evaluate the applicability for food packaging, prepared films were characterised morphological, physical, rheological, mechanical and barrier properties, performed FT-IR, thermal analysis, and finally investigated the antibacterial activity and acute oral toxicity of films. The surface of films presented irregular arrangement with nanoparticles combined in the networks, suggesting the best "CMC to DOM weight ratio" of approximately 1:1 could provide a smooth surface. The films with 2.0 g ZnO-NPs presented antibacterial effects against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria. With the increase of DOM, the antibacterial rate dropped at 400 and 450 µL/mL. The film-forming solutions with higher ZnO-NPs content display shear-thinning properties and liquid-like behaviour. The edible films have a great potential to be used in application in food packaging.
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Antibacterianos/química , Antibacterianos/farmacologia , Carboximetilcelulose Sódica/química , Dioscorea/química , Filmes Comestíveis , Glicerol/química , Óxido de Zinco/química , Escherichia coli/efeitos dos fármacos , Nanocompostos/química , Nanopartículas/química , Staphylococcus aureus/efeitos dos fármacosRESUMO
The biodegradable films, composed of carboxymethyl cellulose (CMC), glycerol, mucilage from Dioscorea opposita (DOM) and Ag nanoparticles, were prepared by a casting method and characterised including colour measurement, solubility, mechanical and water barrier properties etc. to fit the requirements for food packaging. The films were also analysed by fourier transformed infrared spectroscopy, thermal analysis, scanning electron microscopy, and rheometer etc., meanwhile, the antimicrobial activities and acute toxicity of the films were investigated. The addition of Ag nanoparticles and DOM decreased the tensile strength, water solubility and transparency value, while increased the elongation at break. The functional groups and thermal analysis of films presented no significant differences, and the film-forming solutions showed similar shear-thinning properties. Antimicrobial evaluation revealed that the films achieved significant antibaterial effects against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria. The biodegradable films presented a compact and uniform structure with antibacterial properties and without toxicological responses, which has excellent potential to be applied in food packaging.
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Antibacterianos/química , Plásticos Biodegradáveis/química , Dioscorea/química , Nanopartículas Metálicas/química , Adesivos/química , Adesivos/farmacologia , Antibacterianos/farmacologia , Plásticos Biodegradáveis/farmacologia , Carboximetilcelulose Sódica/química , Carboximetilcelulose Sódica/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/patogenicidade , Testes de Sensibilidade Microbiana , Reologia , Prata/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/patogenicidadeRESUMO
The physical properties of the mucilage polysaccharides of Dioscorea opposita (DOMP) were investigated in this study. The monosaccharide and amino acid contents, and molecular weight were determined, and morphology was observed. The rheological and emulsifying properties of different concentrations of DOMP were determined at acidic and basic pH (pH 5.0 and 9.0). The glucose and protein contents were 11.05% and 13.39%, respectively, and the average molecular weight was 9062 Da. The DOMP particles were spheres of 0.18 µm diameter, which aggregated in solution. The viscosity of DOMP decreased gradually with increase in shear rate, which was indicative of pseudoplastic characteristics. DOMP showed relatively better emulsification properties than Konjac glucomannan (KGM). The particle size of DOMP decreased and its emulsifying properties improved under both acidic and basic conditions. These results suggested that DOMP can be used as a natural processing agent for improving the mouth-feel of food.
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Aminoácidos/análise , Dioscorea/química , Mucilagem Vegetal/análise , Polissacarídeos/análise , Emulsificantes/análise , Manipulação de Alimentos , Humanos , Peso Molecular , Monossacarídeos/análise , Tamanho da Partícula , Reologia , ViscosidadeRESUMO
BACKGROUND: The SnRK2s (Plant specific protein kinase) are involved in various biological processes, such as plant defense and environmental challenges. In Arabidopsis, AtSnRK2s regulate the expression of some metal transporters. For example, AtSnRK2.4 plays a role in the regulation of Arabidopsis tolerance to Cd; AtSnRK2.2 and AtSnRK2.3 are involved in Cd uptake and translocation. However, the functions of their homologs, TpSnRK2.10 and TpSnRK2.11 from dwarf Polish wheat are unknown. RESULTS: TpSnRK2.11 encodes a cytoplasmic protein. TpSnRK2.10 and TpSnRK2.11 have different expression patterns at different growth stages. Expression of TpSnRK2.10 increased yeast's sensitivity to Cd; conversely, expression of TpSnRK2.11 enhanced yeast's tolerance to Cd. Overexpression of TpSnRK2.10 or TpSnRK2.11 did not affect Cd sensitivity in Arabidopsis, but significantly increased Cd accumulation in roots and shoots, and Cd translocation from roots to shoots. While, Fe accumulation was significantly increased in roots but decreased in shoots by overexpression of TpSnRK2.10; opposite results were observed in TpSnRK2.11-overexpressing lines. Subcellular distribution analysis found that overexpression of TpSnRK2.10 and TpSnRK2.11 increased Cd concentration in cell wall and organelle fractions of roots and shoots; meanwhile, they also differentially influenced Fe distribution. CONCLUSIONS: These results indicated that TpSnRK2.10 and TpSnRK2.11 are involved in the uptakes and the translocations of Cd and Fe, possibly by regulating the expression of AtNRAMP1 and AtHMA4, and other genes involved in the synthesis of phytochelatins or hemicellolosic polysaccharides.
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Arabidopsis/crescimento & desenvolvimento , Cádmio/metabolismo , Ferro/metabolismo , Proteínas Quinases/metabolismo , Triticum/metabolismo , Adaptação Fisiológica , Arabidopsis/genética , Arabidopsis/metabolismo , Parede Celular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Proteínas Quinases/genética , Triticum/genéticaRESUMO
This study compared the characterisations of polysaccharides from Chinese yam (Dioscorea opposita Thunb.) growing in sandy soil (SSCY), loessial soil (LSCY) and second-year continuous cropping (CCCY). SSCY contained the highest total polysaccharides (36.55%) and 80.19% glucose, CCCY from sandy soil obtained 24.55% polysaccharides with 43.66% glucose, whereas LSCY contained 27.54% total polysaccharides and 7.94% glucose. The results indicated that Dioscorea opposita from sandy soil may obtain higher level of glucose. CCCY increased the galacturonic acids in yams from 7.03% to 26.19%, which may have been caused by the decrease in soil pH due to continuous cropping. The starches of SSCY and CCCY from sandy soil belongs to C-type, whereas the starch of LSCY from loessial soil has the A-type pattern. The results suggested that the two types of soil and continuous cropping change the compounds and contents of yams, which provide valuable evidences for cropping management and allelopathy effects.
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Agricultura/métodos , Dioscorea/química , Polissacarídeos/química , Solo/química , Amilose/análise , Dioscorea/anatomia & histologia , Peso Molecular , Monossacarídeos/análise , Tamanho da Partícula , Polissacarídeos/isolamento & purificação , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Amido/química , ViscosidadeRESUMO
Different intra- or interspecific wheat show different interactions of Cd/Zn. Normally, Zn has been/being widely utilized to reduce the Cd toxicity. In the present study, the DPW seedlings exhibited strong Cd tolerance. Zn and Cd mutually inhibited their uptake in the roots, showed antagonistic Cd/Zn interactions. However, Zn promoted the Cd transport from the roots to shoots, showed synergistic. In order to discover the interactive molecular responses, a transcriptome, including 123,300 unigenes, was constructed using RNA-Sequencing (RNA-Seq). Compared with CK, the expression of 1,269, 820, and 1,254 unigenes was significantly affected by Cd, Zn, and Cd+Zn, respectively. Only 381 unigenes were co-induced by these three treatments. Several metal transporters, such as cadmium-transporting ATPase and plant cadmium resistance 4, were specifically regulated by Cd+Zn. Other metal-related unigenes, such as ABC transporters, metal chelator, nicotianamine synthase (NAS), vacuolar iron transporters (VIT), metal-nicotianamine transporter YSL (YSL), and nitrate transporter (NRT), were regulated by Cd, but were not regulated by Cd+Zn. These results indicated that these transporters participated in the mutual inhibition of the Cd/Zn uptake in the roots, and also participated in the Cd transport, accumulation and detoxification. Meanwhile, some unigenes involved in other processes, such as oxidation-reduction, auxin metabolism, glutathione (GSH) metabolism nitrate transport, played different and important roles in the detoxification of these heavy metals.
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Cd and Zn have been shown to interact antagonistically or synergistically in various plants. In the present study of dwarf polish wheat (DPW)roots, Cd uptake was inhibited by Zn, and Zn uptake was inhibited by Cd, suggesting that Cd and Zn interact antagonistically in this plant. A study of proteomic changes showed that Cd, Zn, and Cd+Zn stresses altered the expression of 206, 303, and 190 proteins respectively. Among these, 53 proteins were altered significantly in response to all these stresses (Cd, Zn, and Cd+Zn), whereas 58, 131, and 47 proteins were altered in response to individual stresses (Cd, Zn, and Cd+Zn, respectively). Sixty-one differentially expressed proteins (DEPs) were induced in response to both Cd and Zn stresses; 33 proteins were induced in response to both Cd and Cd+Zn stresses; and 57 proteins were induced in response to both Zn and Cd+Zn stresses. These results indicate that Cd and Zn induce differential molecular responses, which result in differing interactions of Cd/Zn. A number of proteins that mainly participate in oxidation-reduction and GSH, SAM, and sucrose metabolisms were induced in response to Cd stress, but not Cd+Zn stress. This result indicates that these proteins participate in Zn inhibition of Cd uptake and ultimately cause Zn detoxification of Cd. Meanwhile, a number of proteins that mainly participate in sucrose and organic acid metabolisms and oxidation-reduction were induced in response to Zn stress but not Cd+Zn stress. This result indicates that these proteins participate in Cd inhibition of Zn uptake and ultimately cause the Cd detoxification of Zn. Other proteins induced in response to Cd, Zn, or Cd+Zn stress, participate in ribosome biogenesis, DNA metabolism, and protein folding/modification and may also participate in the differential defense mechanisms.
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Construction as well as characterization of a polish wheat transcriptome is a crucial step to study useful traits of polish wheat. In this study, a transcriptome, including 76,014 unigenes, was assembled from dwarf polish wheat (DPW) roots, stems, and leaves using the software of Trinity. Among these unigenes, 61,748 (81.23%) unigenes were functionally annotated in public databases and classified into differentially functional types. Aligning this transcriptome against draft wheat genome released by the International Wheat Genome Sequencing Consortium (IWGSC), 57,331 (75.42%) unigenes, including 26,122 AB-specific and 2,622 D-specific unigenes, were mapped on A, B, and/or D genomes. Compared with the transcriptome of T. turgidum, 56,343 unigenes were matched with 103,327 unigenes of T. turgidum. Compared with the genomes of rice and barley, 14,404 and 7,007 unigenes were matched with 14,608 genes of barley and 7,708 genes of rice, respectively. On the other hand, 2,148, 1,611, and 2,707 unigenes were expressed specifically in roots, stems, and leaves, respectively. Finally, 5,531 SSR sequences were observed from 4,531 unigenes, and 518 primer pairs were designed.