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Human immunodeficiency virus (HIV) infection is still a global public health issue, and the development of an effective prophylactic vaccine inducing potent neutralizing antibodies remains a significant challenge. This study aims to explore the inflammation-related proteins associated with the neutralizing antibodies induced by the DNA/rTV vaccine. In this study, we employed the Olink chip to analyze the inflammation-related proteins in plasma in healthy individuals receiving HIV candidate vaccine (DNA priming and recombinant vaccinia virus rTV boosting) and compared the differences between neutralizing antibody-positive (nab + ) and -negative(nab-) groups. We identified 25 differentially expressed factors and conducted enrichment and correlation analysis on them. Our results revealed that significant expression differences in artemin (ARTN) and C-C motif chemokine ligand 23 (CCL23) between nab+ and -nab- groups. Notably, the expression of CCL23 was negatively corelated to the ID50 of neutralizing antibodies and the intensity of the CD4+ T cell responses. This study enriches our understanding of the immune picture induced by the DNA/rTV vaccine, and provides insights for future HIV vaccine development.
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Vacinas contra a AIDS , Anticorpos Neutralizantes , Anticorpos Anti-HIV , Infecções por HIV , HIV-1 , Proteômica , Vaccinia virus , Humanos , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Vaccinia virus/imunologia , Vaccinia virus/genética , Anticorpos Anti-HIV/sangue , Anticorpos Anti-HIV/imunologia , HIV-1/imunologia , HIV-1/genética , Adulto , Vacinas contra a AIDS/imunologia , Masculino , Infecções por HIV/imunologia , Vacinas de DNA/imunologia , Feminino , Voluntários Saudáveis , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/administração & dosagem , Plasma/imunologia , Adulto JovemRESUMO
Acidic tea (Camellia sinensis) plantation soil usually suffers from magnesium (Mg) deficiency, and as such, application of fertilizer containing Mg can substantially increase tea quality by enhancing the accumulation of nitrogen (N)-containing chemicals such as amino acids in young tea shoots. However, the molecular mechanisms underlying the promoting effects of Mg on N assimilation in tea plants remain unclear. Here, both hydroponic and field experiments were conducted to analyze N, Mg, metabolite contents, and gene expression patterns in tea plants. We found that N and amino acids accumulated in tea plant roots under Mg deficiency, while metabolism of N was enhanced by Mg supplementation, especially under a low N fertilizer regime. 15N tracing experiments demonstrated that assimilation of N was induced in tea roots following Mg application. Furthermore, weighted gene correlation network analysis (WGCNA) analysis of RNA-seq data suggested that genes encoding glutamine synthetase isozymes (CsGSs), key enzymes regulating N assimilation, were markedly regulated by Mg treatment. Overexpression of CsGS1.1 in Arabidopsis (Arabidopsis thaliana) resulted in a more tolerant phenotype under Mg deficiency and increased N assimilation. These results validate our suggestion that Mg transcriptionally regulates CsGS1.1 during the enhanced assimilation of N in tea plant. Moreover, results of a field experiment demonstrated that high Mg and low N had positive effects on tea quality. This study deepens our understanding of the molecular mechanisms underlying the interactive effects of Mg and N in tea plants while also providing both genetic and agronomic tools for future improvement of tea production.
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Camellia sinensis , Camellia sinensis/genética , Camellia sinensis/metabolismo , Magnésio/metabolismo , Glutamato-Amônia Ligase/genética , Glutamato-Amônia Ligase/metabolismo , Nitrogênio/metabolismo , Fertilizantes , Aminoácidos/metabolismo , Chá/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismoRESUMO
Phosphorus nutrition has been known to influence floral transition in plants for a long time, but the underlying mechanism is unclear. Arabidopsis PHOSPHATE1 (PHO1) plays a critical role in phosphate translocation from roots to shoots, but whether and how it regulates floral transition is unknown. Here, we show that knockout mutation of PHO1 delays flowering under both long-day and short-day conditions. The late flowering of pho1 mutants can be partially rescued by Pi supplementation in rosettes or shoot apices. Grafting assay indicates that the late flowering of pho1 mutants is resulted from impaired phosphate translocation from roots to shoots. Knockout mutation of SPX1 and SPX2, two negative regulators of phosphate starvation response, partially rescues the late flowering of pho1 mutants. PHO1 is epistatic to PHO2, a negative regulator of PHO1, in flowering time regulation. Loss of PHO1 represses the expression of some floral activators, including FT encoding florigen, and induces the expression of some floral repressors in shoots. Genetic analyses indicate that at least jasmonic acid signaling is partially responsible for the late flowering of pho1 mutants. In addition, we find rice PHO1;2, the homology of PHO1, plays a similar role in floral transition. These results suggest that PHO1 integrates phosphorus nutrition and flowering time and could be used as a potential target in modulating phosphorus nutrition-mediated flowering time in plants.
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Aqueous zinc-ion batteries (ZIBs) are widely recognized as highly promising energy storage devices because of their inherent characteristics, including superior safety, affordability, eco-friendliness, and various other benefits. However, the significant corrosion of the zinc metal anode, side reactions occurring between the anode and electrolyte, and the formation of zinc dendrites significantly hinder the practical utilization of ZIBs. Herein, we utilized an electrodeposition method to apply a unique hydrous molybdenum oxide (HMoOx) layer onto the surface of the zinc metal anode, aiming to mitigate its corrosion and side reactions during the process of zinc deposition and stripping. In addition, the HMoOx layer not only improved the hydrophilicity of the zinc anode, but also adjusted the migration of Zn2+, thus facilitating the uniform deposition of Zn2+ to reduce dendrite formation. A symmetrical cell with the HMoOx-Zn anode displayed reduced-voltage hysteresis (80 mV at 2.5 mA/cm2) and outstanding cycle stability after 3000 cycles, surpassing the performance of the uncoated Zn anode. Moreover, the HMoOx-Zn anode coupled with a γ-MnO2 cathode created a considerably more stable rechargeable full battery compared to the bare Zn anode. The HMoOx-Zn||γ-MnO2 full cell also displayed excellent cycling stability with a charge/discharge-specific capacity of 129/133 mAh g-1 after 300 cycles. In summary, this research offers a straightforward and advantageous approach that can significantly contribute to the future advancements in rechargeable ZIBs.
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BACKGROUND: Chenxiang Huaqi tablets (CXHQTs) are a traditional Chinese medicine (TCM) commonly used to treat stomach-related diseases. Currently, the ministerial standards do not provide detailed guidance and regulations on the content determination of CXHQTs, and the reported studies only use individual active components as indicators for determining effective ingredients. OBJECTIVE: The present study aimed to propose a methodology for quality control of CXHQTs based on high-performance liquid chromatography (HPLC) fingerprinting combined with the quantitative analysis of multi-components by single marker (QAMS) method. METHODS: HPLC method was used to determine seven active ingredients and performed fingerprint analysis of CXHQTs. To further process chemometric assessment, technical analysis-model including similarity analysis (SA), hierarchical clustering analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) was set up to differentiate and classify the 20 batches of samples. RESULTS: After comparing the results of QAMS method with the external standard method (ESM), we found there is no significant difference. Besides, the fingerprint of CXHQT was also established. CONCLUSION: HPLC fingerprint combined with the QAMS could be an efficient and selective analysis technique to achieve a qualitative and quantitative evaluation of executing quality processes.
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Medicamentos de Ervas Chinesas , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Medicina Tradicional Chinesa , Controle de Qualidade , ComprimidosRESUMO
The mechanical strength and ionic conductivity of sulfide solid electrolytes have received widespread attention for their application in solid sodium batteries. Herein, first-principles calculations are used to determine the properties, including the electronic, mechanical and ionic transport properties, of Na3PS4 sulfide solid electrolytes doped with low and high Ca ion concentrations. Our theoretical results demonstrate that low Ca ion concentrations can be easily doped in tetragonal and cubic phases (t-Na3PS4 and c-Na3PS4) and create a suitable number of Na vacancies based on the formation energy analysis. Furthermore, the calculated density of states and charge density differences indicate that the surrounding electronic environment is changed, and Ca-S ionic bonds are formed in Na3PS4 with Ca-doping. In addition, the improved ductility and mechanical strength of c-Na3PS4 and t-Na3PS4 achieved by low-concentration Ca doping may help suppress dendritic growth and electrode deformation. Finally, sodium ion migration in Ca-doped Na3PS4 is described with the aid of the CI-NEB method, and it is found that the migration energy barriers are less than those of pure Na3PS4, which suggests that the sodium ion conductivity can be effectively improved by doping with low Ca2+ concentrations. The present work improves the understanding of the influence of doping on the performance of solid electrolytes and provides a feasible framework for the future design of high-performance solid electrodes.
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Herein, we report an organic photoredox-catalyzed hydroazidation of trifluoromethyl alkenes with user-friendly trimethylsilyl azide (TMSN3), enabling a direct access to a broad range of valuable ß-CF3-azides with exclusive selectivity under mild reaction conditions. The synthetic utility of this reaction was demonstrated by the late-stage modification of complex drug derivatives, scale-up of the reaction and diverse further derivatizations of the ß-CF3-azide product.
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Herein, we report a photoredox enabled defluorinative benzylation of trifluoromethyl alkenes with readily available alkylarenes, which provides convenient access to a series of structurally valuable benzylated gem-difluoroalkenes under mild reaction conditions. The synthetic value of this protocol has been demonstrated by the transformations of several substrates bearing drug moieties, gram-scale reactions, and various further derivatizations of the gem-difluoroalkene products. The preliminary mechanistic investigations suggest a reaction pathway with rate-determining benzyl C-H bond cleavage of toluene followed by benzylic radical formation.
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Early cancer diagnosis increases therapy efficiency and saves huge medical costs. Traditional blood-based cancer markers and endoscopy procedures demonstrate limited capability in the diagnosis. Reliable, non-invasive, and cost-effective methods are in high demand across the world. Worm-based diagnosis, utilizing the chemosensory neuronal system of C. elegans, emerges as a non-invasive approach for early cancer diagnosis with high sensitivity. It facilitates effectiveness in large-scale cancer screening for the foreseeable future. Here, we review the progress of a unique route of early cancer diagnosis based on the chemosensory neuronal system of C. elegans. We first introduce the basic procedures of the chemotaxis assay of C. elegans: synchronization, behavior assay, immobilization, and counting. Then, we review the progress of each procedure and the various cancer types for which this method has achieved early diagnosis. For each procedure, we list examples of microfluidics technologies that have improved the automation, throughput, and efficiency of each step or module. Finally, we envision that microfluidics technologies combined with the chemotaxis assay of C. elegans can lead to an automated, cost-effective, non-invasive early cancer screening technology, with the development of more mature microfluidic modules as well as systematic integration of functional modules.
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The membrane-proximal external region (MPER) represents a highly conserved region of the Human Immunodeficiency Virus (HIV) envelope glycoprotein (env) targeted by several broadly neutralizing antibodies (bnAbs). In this study, we employed single genome amplification to amplify 34 full-length env sequences from the 2005 plasma sample of CBJC504, a chronic HIV-1 clade B infected individual. We identified three amino acid changes (N671S, D674N, and K677R) in the MPER. A longitudinal analysis revealed that the proportion of env sequences with MPER mutations increased from 26.5 % in 2005 to 56.0 % in 2009, and the sequences with the same mutation clustered together. Nine functional pseudoviruses were generated from the 34 env sequences to examine the effect of these mutations on neutralizing activity. Pseudoviruses carrying N674 or R677 mutations demonstrate increased sensitivity to autologous plasma and monoclonal antibodies 2F5, 4E10, and 10E8. Reverse mutations were performed in env including N674, R677, D659, and S671/N677 mutations, to validate the impact of the mutations on neutralizing sensitivity. Neutralization assays indicated that the N671S mutation increased neutralization sensitivity to 2F5 and 10E8. The amino acid R at position 677 increased viral resistance to 10E8, whereas N enhanced viral resistance to 4E10 and 10E8. It has been proposed that critical amino acids in the extra-MPER and the number of potential N-like glycosylation sites (PNGSs) in the V1 loop may have an impact on neutralizing activity. Understanding the mutations and evolution of MPER in chronically infected patients with HIV-1 is crucial for the design and development of vaccines that trigger bnAbs against MPER.
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Substituição de Aminoácidos , Anticorpos Neutralizantes , Anticorpos Anti-HIV , Infecções por HIV , HIV-1 , Testes de Neutralização , Produtos do Gene env do Vírus da Imunodeficiência Humana , Humanos , HIV-1/genética , HIV-1/imunologia , Anticorpos Neutralizantes/imunologia , Infecções por HIV/virologia , Infecções por HIV/imunologia , Anticorpos Anti-HIV/imunologia , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia , Estudos LongitudinaisRESUMO
Tea polysaccharide (TPS) is a bioactive compound extracted from tea. It has raised great interest among researchers due to its bioactivity. However, few studies focused on the diversity of TPS in its compositions and antioxidant activity. This study collected 140 different tea varieties from four tea germplasm gardens in China, and their TPSs in tea shoots were extracted. The extraction efficiency, composition contents, including neutral sugar, uronic acid, protein, and tea polyphenols, and the scavenging abilities of hydroxyl radical (·OH) and superoxide radical (O2-·) of 140 TPSs were determined and analyzed. The results showed significant differences in the compositions and antioxidant activities of TPS extracted from different tea varieties. By applying hierarchical clustering analysis (HCA), we selected nine tea varieties with high TPS extraction efficiency and 26 kinds of TPS with high antioxidant capacity.
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Epigallocatechin-3-gallate (EGCG), a flavoured and healthy compounds in tea, is affected by the ecological factors. However, the biosynthetic mechanisms of EGCG in response to the ecological factors remian unclear. In this study, a response surface method with a Box-Behnken design was used to investigate the relationship between EGCG accumulation and ecological factors; further, integrative transcriptome and metabolome analyses were performed to explore the mechanism underlying EGCG biosynthesis in response to environmental factors. The optimal environmental conditions obtained for EGCG biosynthesis were as follows: 28â, 70 % relative humidity of the substrate, and 280 µmol·m-2·s-1 light intensity; the EGCG content was increased by 86.83 % compared to the control (CK1). Meanwhile, the order of EGCG content in response to the interaction of ecological factors was as follows: interaction of temperature and light intensity > interaction of temperature and relative humidity of the substrate > interaction of light intensity and relative humidity of the substrate, indicating that temperature was the dominant ecological factors. EGCG biosynthesis in tea plants was found to be comprehensively regulated by a series of structural genes (CsANS, CsF3H, CsCHI, CsCHS, and CsaroDE), miRNAs (miR164, miR396d, miR5264, miR166a, miR171d, miR529, miR396a, miR169, miR7814, miR3444b, and miR5240), and transcription factors (MYB93, NAC2, NAC6, NAC43, WRK24, bHLH30, and WRK70); further, the metabolic flux was regulated and converted from phenolic acid to the flavonoid biosynthesis pathway based on accelerated consumption of phosphoenolpyruvic acid, d-erythrose-4-phosphate, and l-phenylalanine in response to ambient changes in temperature and light intensity. Overall, the results of this study reveal the effect of ecological factors on EGCG biosynthesis in tea plants, providing novel insights for improving tea quality.
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Camellia sinensis , Camellia sinensis/química , Transcriptoma , Metaboloma , Chá/químicaRESUMO
Catechins are the major flavor substances in teas, which have a variety of health effects; however, high catechin and high sensory quality are a pair of contradictions that are difficult to coordinate. To explore the processing procedure with high catechins and high sensory quality, a single-factor processing experiment was carried out over the processing production of oolong tea. Combined with orthogonal partial least square discriminant analysis (OPLS-DA), correlation analysis, and principal component analysis (PCA), the optimal production procedure for oolong tea is as follows: red light withering for 8 h, leaf rotating for 10 min with a total standing time for 8 h, drum roasting for 5 min at 290 °C, low-temperature rolling (flattening at 4 °C for 5 min, without pressure for 1 min and under pressure for 5 min), microwave drying (800 W for 7.5 min). This study demonstrates a significant increase in the retention of catechins, which contributes to the mellow and brisk tastes of oolong tea, addressing the challenge of catechin content and sensory quality. Our study provides a novel insight into the relationship between the oolong tea processing and flavor formation.
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As the main flavor components of tea, the contents of epigallocatechin-3-gallate (EGCG), theanine and caffeine are regulated by ambient temperature. However, whether the biosynthesis of EGCG, theanine and caffeine in response to temperature is regulated by endogenous hormones and its mechanism is still unclear. In this study, tea cuttings cultivated in the phytotron which treated at different temperatures 15â, 20â, 25â and 30â, respectively. The UPLC and ESI-HPLC-MS/MS were used to determine the contents of EGCG, theanine, caffeine and the contents of phytohormones in one leaf and a bud. The results showed that indoleacetic acid (IAA), gibberellin 1(GA1) and gibberellin 3 (GA3) were significantly correlated with the content of EGCG; Jasmonic acid (JA), jasmonate-isoleucine (JA-Ile) and methyl jasmonate (MeJA) were strongly correlated with theanine content; IAA, GA1 and gibberellin 4 (GA4) were significantly correlated with caffeine content at different temperatures. In order to explore the internal intricate relationships between the biosynthesis of these three main taste components, endogenous hormones, and structural genes in tea plants, we used multi-omics and multidimensional correlation analysis to speculate the regulatory mechanisms: IAA, GA1 and GA3 up-regulated the expressions of chalcone synthase (CsCHS) and trans-cinnamate 4-monooxygenase (CsC4H) mediated by the signal transduction factors auxin-responsive protein IAA (CsIAA) and DELLA protein (CsDELLA), respectively, which promoted the biosynthesis of EGCG; IAA, GA3 and GA1 up-regulated the expression of CsCHS and anthocyanidin synthase (CsANS) mediated by CsIAA and CsDELLA, respectively, via the transcription factor WRKY DNA-binding protein (CsWRKY), and promoted the biosynthesis of EGCG; JA, JA-Ile and MeJA jointly up-regulated the expression of carbonic anhydrase (CsCA) and down-regulated the expression of glutamate decarboxylase (CsgadB) mediated by the signal transduction factors jasmonate ZIM domain-containing protein (CsJAZ), and promoted the biosynthesis of theanine; JA, JA-Ile and MeJA also jointly inhibited the expression of CsgadB mediated by CsJAZ via the transcription factor CsWRKY and AP2 family protein (CsAP2), which promoted the biosynthesis of theanine; IAA inhibited the expression of adenylosuccinate synthase (CspurA) mediated by CsIAA via the transcription factor CsWRKY; GA1 and gibberellin 4 (GA4) inhibited the expression of CspurA mediated by CsDELLA through the transcription factor CsWRKY, which promoted the biosynthesis of caffeine. In conclusion, we revealed the underlying mechanism of the biosynthesis of the main taste components in tea plant in response to temperature was mediated by hormone signal transduction factors, which provided novel insights into improving the quality of tea.
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Bone drilling is an important procedure in medical orthopedic surgery and it is inevitable that heat will be generated during the drilling process and higher temperatures can cause thermal damage to the bone tissue near the drilled hole. Therefore, the capability to obtain the cortical bone drilling temperature distribution area can have great significance for medical bone surgery. Based on the theory of heat transfer, a predictive model for cortical bone drilling temperature distribution was established. The energy distribution coefficient in cortical bone drilling was derived, based on conjugate gradient inversion. A cortical bone drilling experiment platform was built to verify the temperature distribution prediction model. The results show that the model of cortical bone drill temperature distribution could predict accurately the drilling temperature distribution, both for different depths and for different radial distances. Additionally, the effects of different drilling conditions (spindle speed, feed rate, drill diameter) on the temperature of drilling cortical bone were considered.
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Osso e Ossos , Procedimentos Ortopédicos , Osso Cortical/cirurgia , Temperatura Alta , TemperaturaRESUMO
Enterotoxigenic Escherichia coli (ETEC) is a common enteric pathogen that causes diarrhoea in humans and animals. Lactobacillus rhamnosus LB1 (formerly named Lactobacillus zeae LB1) has been shown to reduce ETEC infection to Caenorhabditis elegans and Salmonella burden in pigs. This study was to evaluate the effect of L. rhamnosus LB1 on the gut health of lactating piglets that were challenged with ETEC. Six-four piglets at 7 days of age were equally assigned into 8 groups (8 piglets per group): 1) control group (basal diet, phosphate buffer saline); 2) CT group (basal diet + 40 mg/kg colistin); 3) LL group (basal diet + 1 × 107 CFU/pig/day LB1); 4) HL group (basal diet + 1 × 108 CFU/pig/day LB1); 5) ETEC group: (basal diet + ETEC challenged); 6) CT + ETEC group (basal diet + CT + ETEC); 7) LL + ETEC group (basal diet + 1 × 107 CFU/pig/day LB1 + ETEC); 8) HL + ETEC group (basal diet + 1 × 108 CFU/pig/day LB1 + ETEC). The trial lasted ten days including 3 days of adaptation. Several significant interactions were found on blood parameters, intestinal morphology, gene, and protein expression. ETEC infection disrupted the cell structure and biochemical indicators of blood, undermined the integrity of the intestinal tract, and induced oxidative stress, diarrhoea, intestinal damage, and death of piglets. The supplementation of L. rhamnosus LB1 alleviated ETEC's adverse effects by reducing pig diarrhoea, oxidative stress, and death, modulating cell structure and biochemical indicators of blood, improving the capacity of immunity and anti-oxidation stress of pigs, and restoring their intestinal integrity. At the molecular level, the beneficial effects of L. rhamnosus LB1 appeared to be mediated by regulating functional related proteins (including HSP70, Caspase-3, NLRP3, AQP3, and AQP4) and genes (including RPL4, IL-8, HP, HSP70, Mx1, Mx2, S100A12, Nrf2, GPX2 and ARG1). These results suggest that dietary supplementation of L. rhamnosus LB1 improved the intestinal functions and health of piglets.
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Escherichia coli Enterotoxigênica , Infecções por Escherichia coli , Lacticaseibacillus rhamnosus , Doenças dos Suínos , Animais , Caenorhabditis elegans , Infecções por Escherichia coli/prevenção & controle , Infecções por Escherichia coli/veterinária , Feminino , Imunidade , Lactação , Lactobacillus , Estresse Oxidativo , SuínosRESUMO
Eco-friendly cellulose acetate (CA) monolith with novel hierarchical micro/nano-porous structure was successfully fabricated via a simple thermally impacted nonsolvent induced phase separation (TINIPS) method. Based on the unique three-dimensional (3D) hierarchical porous structure, CA monolith revealed a high porosity (92.1%), excellent hydrophobicity (water contact angle of 147°) and superoleophilicity (oil contact angle of 0°). As a result, the porous monolith could selectively and efficiently adsorb various oils and organic solvents from oil/water mixtures with high saturation adsorption capacity (Qm) of 6.59-15.03â¯gâ¯g-1. Besides, the monolith exhibited outstanding environmental stability in different pH (1-14), temperature (0-70⯰C) and turbulent environments with almost unchanged hydrophobicity and Qm. Besides, CA monolith also showed a continuous oil/water separation ability to purify the polluted water by using a pump-assisted system, revealing a great potential for controlling ocean oil pollution.
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Celulose/análogos & derivados , Nanoestruturas/química , Óleos/química , Poluentes Químicos da Água/química , Purificação da Água , Água/química , Adsorção , Celulose/química , Interações Hidrofóbicas e Hidrofílicas , Poluição por Petróleo , Porosidade , SolventesRESUMO
A gas chromatographic fingerprint combined with chemical pattern recognition was successfully developed and applied to assess the quality consistency of 20 Chenxianghuaqi tablets. Volatile components from the 20 Chenxianghuaqi tablets were extracted with ethanol under ultrasonic conditions. n-Octadecane was used as the internal standard to calculate the amounts of the three main components and to confirm the relative peak areas of the other components. Gas chromatographic fingerprints of the 20 Chenxianghuaqi tablets were established. There were 11 common peaks in the fingerprints, and the similarity of each batch of samples was obtained. Ten common peaks were identified by gas chromatography-mass spectrometry, with reference comparison. The obtained fingerprints were used for the chemical pattern recognition, including hierarchical cluster analysis and the principal component analysis. Different batches of Chenxianghuaqi tablets could be differentiated effectively, and major markers that led to differences among the sample batches were identified. The method proposed in this study is comprehensive and reliable, and it can be used as a valuable reference to evaluate and control the quality of Chenxianghuaqi tablets.
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Medicamentos de Ervas Chinesas/análise , Controle de Qualidade , Cromatografia Gasosa-Espectrometria de Massas , ComprimidosRESUMO
Enterotoxigenic Escherichia coli (ETEC) is primary pathogenic bacteria of piglet diarrhea, over two thirds of piglets diarrhea caused by ETEC are resulted from STa-producing ETEC strains. This experiment was conducted to construct the recombinant E. coli expressing STa and study the injury and mechanism of recombinant E. coli expressing STa on 7 days old piglets colon. Twenty-four 7 days old piglets were allotted to four treatments: control group, STa group (2 × 109 CFU E. coli LMG194-STa), LMG194 group (2 × 109 CFU E. coli LMG194) and K88 group (2 × 109 CFU E. coli K88). The result showed that E. coli infection significantly increased diarrhea rates; changed DAO activity in plasma and colon; damaged colonic mucosal morphology including crypt depth, number of globet cells, density of lymphocytes and lamina propria cell density; substantially reduced antioxidant capacity by altering activities of GSH-Px, SOD, and TNOS and productions of MDA and H2O2; obviously decreased AQP3, AQP4 and KCNJ13 protein expression levels; substantially altered the gene expression levels of inflammatory cytokines. Conclusively, STa group had the biggest effect on these indices in four treatment groups. These results suggested that the recombinant strain expressed STa can induce piglets diarrhea and colonic morphological and funtional damage by altering expression of proteins connect to transportation function and genes associated with intestinal injury and inflammatory cytokines.
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Toxinas Bacterianas/metabolismo , Colo/microbiologia , Diarreia/veterinária , Escherichia coli Enterotoxigênica/metabolismo , Enterotoxinas/metabolismo , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/metabolismo , Animais , Toxinas Bacterianas/genética , Colo/metabolismo , Colo/patologia , Diarreia/microbiologia , Diarreia/patologia , Escherichia coli Enterotoxigênica/genética , Enterotoxinas/genética , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Proteínas de Escherichia coli/genética , Expressão Gênica , Glutationa Peroxidase/metabolismo , Peróxido de Hidrogênio/metabolismo , Malondialdeído/metabolismo , Óxido Nítrico Sintase/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Superóxido Dismutase/metabolismo , SuínosRESUMO
Aflatoxin B1 (AFB1) is one of the most hazardous mycotoxins contamination in food and feed products, which leads to hepatocellular carcinoma in humans and animals. In the present study, we isolated and characterized an AFB1 degrading bacteria CG1061 from chicken cecum, exhibited an 93.7% AFB1 degradation rate by HPLC. 16S rRNA gene sequence analysis and a multiplex PCR experiment demonstrated that CG1061 was a non-pathogenic Escherichia coli. The culture supernatant of E. coli CG1061 showed an 61.8% degradation rate, whereas the degradation rates produced by the intracellular extracts was only 17.6%, indicating that the active component was constitutively secreted into the extracellular space. The degradation rate decreased from 61.8 to 37.5% when the culture supernatant was treated with 1 mg/mL proteinase K, and remained 51.3% when that treated with 100°C for 20 min. We postulated that AFB1 degradation was mediated by heat-resistant proteins. The content of AFB1 decreased rapidly when it was incubated with the culture supernatant during the first 24 h. The optimal incubation pH and temperature were pH 8.5 and 55°C respectively. According to the UPLC Q-TOF MS analysis, AFB1 was bio-transformed to the product C16H14O5 and other metabolites. Based on the results of in vitro experiments on chicken hepatocellular carcinoma (LMH) cells and in vivo experiments on mice, we confirmed that CG1061-degraded AFB1 are less toxic than the standard AFB1. E. coli CG1061 isolated from healthy chicken cerum is more likely to colonize the animal gut, which might be an excellent candidate for the detoxification of AFB1 in food and feed industry.