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BACKGROUND: Studies have shown that fine particulate matter (PM2.5) remains a significant problem in developing countries and plays a critical role in the onset and progression of respiratory illnesses. Circular RNAs (circRNAs) are involved in many pathophysiological processes,but their relationship to PM2.5 pollution is largely unexplored. OBJECTIVES: To elucidate the functional role of hsa_circ_0000992 in PM2.5-induced inflammation in a human bronchial epithelial cell line (16HBE) and to clarify whether the competing endogenous RNA (ceRNA) mechanism is involved in the interrelationships between hsa_circ_0000992 and hsa-miR-936 and the inflammatory signaling pathways. METHODS: Detection of inflammatory factors in 16HBE cells exposed to PM2.5 by RT-qPCR and ELISA.High throughput sequencing and bioinformatics analysis methods were used to screen circRNA.The bioinformatics analysis method western blotting and dual-luciferase reporter gene system were used to verify mechanisms associated with circRNA. RESULTS: PM2.5 cause inflammation in the 16HBE cells. High throughput sequencing and RT-qPCR result revealed that the expression of hsa_circ_0000992 was markedly up-regulated in 16HBE exposed to PM2.5. The binding sites between hsa_circ_0000992 and hsa-miR-936 was confirmed by dual-luciferase reporter gene system.Western blotting and RT-qPCR showed that hsa_circ_0000992 can interact with hsa-miR-936 to regulate AKT serine/threonine kinase 3(AKT3),thereby activating the PI3K/AKT pathway and ultimately promoting the expression of interleukin (IL)- 1ß and IL-8. CONCLUSION: PM2.5 can induce the inflammatory response in 16HBE cells by activating the PI3K/AKT pathway. The expression of hsa_circ_0000992 increased when PM2.5 stimulated 16HBE cells,and the circRNA could then regulate the inflammatory response.Hsa_circ_0000992 regulates the hsa-miR-936/AKT3 axis through the ceRNA mechanism,thereby activating the PI3K/AKT signaling pathway,increasing the expression of cellular inflammatory factors,and promoting PM2.5-induced respiratory inflammation.
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MicroARNs , Humanos , MicroARNs/genética , MicroARNs/metabolismo , ARN Circular/genética , ARN Circular/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Células Epiteliales/metabolismo , Material Particulado/toxicidad , Inflamación/inducido químicamente , Inflamación/genética , LuciferasasRESUMEN
The melanocortin 4 receptor (MC4R) is a G protein-coupled transporter that mediates the regulation of thyroid hormones and leptin on energy balance and food intake. However, the mechanisms of transcriptional regulation of Mc4r by thyroid hormone and leptin in fish have been rarely reported. The messenger RNA expression of Mc4r gene was significantly higher in brain than those in other tissues of mandarin fish. We analyzed the structure and function of a 2029 bp sequence of Mc4r promoter. Meanwhile, overexpression of NKX2.1 and incubation with leptin significantly increased Mc4r promoter activity, but triiodothyronine showed the opposite effect. In addition, mutations in the NKX2.1 binding site abolished not only the activation of Mc4r promoter activity by leptin but also the inhibitory effect of thyroid hormones on Mc4r promoter activity. In summary, these results suggested that thyroid hormones and leptin might regulate the transcriptional expression of Mc4r through NKX2.1.
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Peces , Genes Homeobox , Leptina , Animales , Proteínas de Unión al ADN/genética , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Peces/genética , Peces/metabolismo , Leptina/genética , Leptina/farmacología , Regiones Promotoras Genéticas/genética , Receptor de Melanocortina Tipo 4/genética , Receptor de Melanocortina Tipo 4/metabolismo , Hormonas Tiroideas , Factor Nuclear Tiroideo 1/genética , Factor Nuclear Tiroideo 1/metabolismo , Humanos , Células HEK293RESUMEN
At present, studies have found that latent Epstein-Barr virus (EBV) infection is associated with a variety of human tumours, and a vaccine is not available in this field. In this research, RT-PCR was used to obtain BZLF1 (immediately expressed early antigen Z) and LMP2 (latent membrane protein 2) cDNA from EBV. A ZLP2 fusion gene containing a linker sequence that encoded the polypeptide (Gly4Ser)3 was obtained using the sequence splicing overlap extension method. Then, ZLP2 was inserted into pMV261 cells, and the recombinant plasmid pMV-ZLP2 was transformed into BCG competent cells. After EB virus-positive tumour cell (NPRC18) cancer models were established with C57BL/6 J mice, tumour weight, tumour formation time and mouse survival conditions were analyzed, and flow cytometry was used to analyze the quantities of CD8 + and CD4 + T cells. HE staining was used to detect and analyze lymphocyte infiltration, and statistical analysis was used to analyze the immunological effect of recombinant BCG (rBCG). Compared with the control group, rBCG could significantly prolong the survival time of mice, slow tumour growth and delay tumour formation time. Recombinant BCG exhibits an obvious immune effect in mice and an inhibitory effect on EBV-positive cancer.Key points⢠AZLP2 fusion gene with BZLF1 and LMP2 of EB virus was constructed.⢠ZLP2 fusion gene was expressed with rBCG.⢠rBCG with ZLP2 has an obvious effect on EBV-positive cancer.
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Infecciones por Virus de Epstein-Barr , Neoplasias , Animales , Vacuna BCG , Linfocitos T CD4-Positivos , Herpesvirus Humano 4/genética , Ratones , Ratones Endogámicos C57BLRESUMEN
The improvement of the harvest period standards is critical in the quality control of Chinese medicinal materials. The present study statistically analyzed the harvest period standards of plant medicinal materials in the 2020 edition of Chinese Pharmacopoeia(Vol.â ) and put forward the existing problems and suggestions based on herbal records and modern research to provide references for the improvement of the standards. According to the statistical analysis, in 499 types of plant medicinal materials, harvest period standards are recorded under 486 types, accounting for 97.4%, and are lacking in the remaining. Only one medicinal material(Stellariae Radix) is recorded with the standard of the harvest year. The standards of the harvest season and phenological period are recorded under 233 types, accounting for 46.7%. For 237 types, only harvest season is specified, accounting for 47.5%, and for 15 types, only harvest phenological period is specified, accounting for 3.0%. Among 222 types mainly derived from cultivation and 51 types from wild resources and cultivation, only 11 types are recorded with harvest period of cultivated products. Only Stellariae Radix is recorded with the harvest period standards for the wild and cultivated products separately. The harvest period standards of plant medicinal materials with different medicinal parts have certain rules to follow. The main problems about the harvest period standards are discovered. Specifically, no harvest period standards are recorded under 13 types of plant medicinal materials. Almost all perennial cultivated medicinal materials are not recorded with harvest year standard. No phenological period standard is found under 250 types of plant medicinal materials. There is no clear distinction between the harvest period standards of cultivated and wild products. The evidence for harvest period standards of 26 types of plant medicinal materials that can be harvested all year round is insufficient. As a result, it is proposed to strengthen basic research in response to the above-mentioned problems and improve the harvest period standards as soon as possible to ensure the quality of Chinese medicinal materials.
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Medicamentos Herbarios Chinos , Plantas Medicinales , Control de Calidad , China , Medicamentos Herbarios Chinos/normas , Medicina Tradicional China , Farmacopeas como AsuntoRESUMEN
With annual Salvia miltiorrhiza seedlings as experimental material, using "3414" optimal regression design recommended by the Ministry of Agriculture and regularly watered with nutrient solution, through the dynamic sampling of S. miltiorrhiza in different growing stages, and the growth index, dry weight of plant root and content of active components were measured. The potted experiments were applied to study the effects of different nitrogen and phosphorus ratios on the growth, dry matter accumulation and accumulation of active components of S. miltiorrhiza, in order to explore a compatible fertilization method of nitrogen and phosphorus ratio that are suitable for production and quality of S. miltiorrhiza. The results reported as followsï¼â High concentrations of nitrogen fertilizer was beneficial to dry matter accumulation of S. miltiorrhiza aerial parts, and low concentration of nitrogen fertilizer transferred the dry matter accumulation to underground, and N1P1 could make the transfer ahead of time;â¡Regression analysis showed that in the early growth stage (before early July), we could use the nitrogen and phosphorus as basic fertilizer at a concentration of 1.521,0.355 gâ¢L⻹ respectively to promote the growth of S. miltiorrhiza and at a concentration of 2.281,0.710 gâ¢L⻹ respectively to promote the dry matter accumulation of root (after mid-August);â¢Five kinds of active components of S. miltiorrhiza decreased with the increase of nitrogen concentration, and increased with the increase of the concentration of phosphate fertilizer. Nitrogenous fertilizer, phosphate fertilizer in N-P=2â¶3 ratio was more suitable for the accumulation of salvianolic acids, in N-P=1â¶2 ratio was more suitable for the accumulation of tanshinone.
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Fertilizantes , Nitrógeno/química , Fósforo/química , Salvia miltiorrhiza/crecimiento & desarrollo , Raíces de Plantas/crecimiento & desarrollo , Salvia miltiorrhiza/químicaRESUMEN
The PstICL1 gene, which encodes isocitrate lyase, a key enzyme in the glyoxylate cycle, was cloned and characterized in the biotrophic wheat pathogen Puccinia striiformis f. sp. tritici (Pst). Expression analyses of PstICL1 exhibited high levels of transcripts in ungerminated urediniospores, which showed low isocitrate lyase enzyme activity. In planta, PstICL1 expression was continuously down-regulated upon germination. During the later stages of the infection of wheat, the level of PstICL1 expression was extremely low. The function of PstICL1 was identified via mutant complementation. The expression of PstICL1 in Saccharomyces cerevisiae can complement the defects of the â³ICL mutant. Using 3-nitropropionate, we observed that inactivation of isocitrate lyase greatly reduced the germination rate of urediniospores, indicating that PstICL1 plays a key role during Pst germination. Furthermore, analysis of lipid bodies revealed that lipid components continuously enter the germ tube from the urediniospore cell during germ tube elongation. Moreover, during this period, the lipid contents continuously decreased, and the total carbohydrates markedly increased, demonstrating that the lipids are being converted into carbohydrates. These results suggest that PstICL1 is required for Pst germination.
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Basidiomycota/crecimiento & desarrollo , Germinación , Isocitratoliasa/genética , Isocitratoliasa/metabolismo , Basidiomycota/enzimología , Clonación Molecular , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación de la Expresión Génica de las Plantas , Nitrocompuestos/farmacología , Filogenia , Propionatos/farmacología , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genéticaRESUMEN
The fungus Puccinia striiformis f. sp. tritici (Pst), the causal agent of wheat stripe rust, is an obligate biotrophic basidiomycete. Many studies have found that myosins play important roles during fungal growth and propagation. However, there are few reports on the myosins of Pst. In this study, we cloned and obtained the myosin light chain gene PsMLC1 from Pst and characterized its expression. Furthermore, the function of PsMLC1 was identified by mutant complementation. As a result, we found that expression of PsMLC1 in Schizosaccharomyces pombe mostly complemented the defects of the cdc4 mutant, indicating that PsMLC1 belongs to the myosin light chain family member. Expression studies showed that the transcript levels of PsMLC1 little changed before 24 h post inoculation then was suddenly down-regulated during Pst infection of wheat. By using ML-7, we observed that inactivity of PsMLC1 greatly reduced the germination rate of urediniospores. These results suggest that PsMLC1 is essential for the early stages of Pst infection of wheat but unnecessary for the later stages of infection. This work elucidates the function of the myosins in Pst and may provide some theoretical basis for controlling strip rust.
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Basidiomycota/genética , Cadenas Ligeras de Miosina/genética , Basidiomycota/aislamiento & purificación , Clonación Molecular , Eliminación de Gen , Expresión Génica , Perfilación de la Expresión Génica , Prueba de Complementación Genética , Enfermedades de las Plantas/microbiología , Schizosaccharomyces/genética , Triticum/microbiologíaRESUMEN
The beneficial effect of social interaction in mitigating the incidence of post-stroke depression (PSD) and ameliorating depressive symptoms has been consistently demonstrated through preclinical and clinical studies. However, the underlying relationship with oxytocin requires further investigation. In light of this, the present study aimed to explore the protective effect of pair housing on the development of PSD and the potential relationship with oxytocin receptors. The PSD model was induced by middle cerebral artery occlusion (MCAO) for 50 min, followed by 4-week isolated housing and restrained stress. Subsequently, each mouse in the pair-housing group (PH) was pair-housed with an isosexual healthy partner. Another group was continuously administrated fluoxetine (10 mg/Kg, i.p, once a day) for 3 weeks. To elucidate the potential role of oxytocin, we subjected pair-housed PSD mice to treatment with an oxytocin receptor (OXTR) antagonist (L368,889) (5 mg/Kg, i.p, once a day) for 3 weeks. At 31 to 32 days after MCAO, anxiety- and depressive-like behaviors were assessed using sucrose consumption, forced swim test, and tail-suspension test. The results showed that pair housing significantly improved post-stroke depression to an extent comparable to that of fluoxetine treatment. Furthermore, pair housing significantly decreased corticosterone in serum, increasing OXT mRNA expression in the hypothalamus. Treatment with L368,889 essentially reversed the effect of pair housing, with no discernible sex differences apart from changes in body weight. Pair housing increased hippocampal serotonin (5-HT), but treatment with L368,889 had no significant impact. Additionally, pair housing effectively reduced the number of reactive astrocytes and increased Nissl's body in the cortex and hippocampal CA3 regions. Correspondingly, treatment with L368,889 significantly reversed the changes in the Nissl's body and reactive astrocytes. Moreover, pair housing downregulated mRNA levels of TNF-α, IL-1ß, and IL-6 in the cortex caused by PSD, which was also reversed by treatment with L368,889. In conclusion, pair housing protects against the development of PSD depending on OXT and OXTR in the brain, with no significant divergence based on sex. These findings provide valuable insights into the potential of social interaction and oxytocin as therapeutic targets for PSD. Further research into the underlying mechanisms of these effects may contribute to the development of novel treatments for PSD.
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Canfanos , Depresión , Modelos Animales de Enfermedad , Fluoxetina , Piperazinas , Receptores de Oxitocina , Animales , Receptores de Oxitocina/metabolismo , Masculino , Depresión/etiología , Depresión/metabolismo , Ratones , Fluoxetina/farmacología , Infarto de la Arteria Cerebral Media/complicaciones , Infarto de la Arteria Cerebral Media/psicología , Vivienda para Animales , Oxitocina/farmacología , Oxitocina/metabolismo , Ratones Endogámicos C57BL , Accidente Cerebrovascular/complicaciones , Accidente Cerebrovascular/psicología , Conducta Animal/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/efectos de los fármacosRESUMEN
Mycobacterium intracellulare-caused pulmonary infections have mostly been reported in immunocompromised hosts, while cutaneous M. intracellulare infections are rare. We describe here an immunocompetent patient with cutaneous lesions due to M. intracellulare, which was diagnosed by acid-fast staining, in vitro culture, histopathology, and PCR-restriction fragment length polymorphism analysis and gene sequencing of heat-shock protein (hsp) 65 and 16S rDNA genes. In vitro susceptibility testing was also carried out and the patient was successfully treated with clarithromycin, rifampicin, and ethambutol.
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Inmunocompetencia , Complejo Mycobacterium avium/aislamiento & purificación , Infección por Mycobacterium avium-intracellulare/microbiología , Enfermedades Cutáneas Bacterianas/microbiología , Piel/microbiología , Adulto , Antituberculosos/uso terapéutico , Proteínas Bacterianas/genética , Chaperonina 60/genética , Claritromicina/uso terapéutico , ADN Bacteriano/análisis , Quimioterapia Combinada , Etambutol/uso terapéutico , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Complejo Mycobacterium avium/efectos de los fármacos , Complejo Mycobacterium avium/genética , Infección por Mycobacterium avium-intracellulare/tratamiento farmacológico , Infección por Mycobacterium avium-intracellulare/inmunología , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Ribotipificación , Rifampin/uso terapéutico , Piel/efectos de los fármacos , Piel/inmunología , Piel/patología , Enfermedades Cutáneas Bacterianas/tratamiento farmacológico , Enfermedades Cutáneas Bacterianas/inmunologíaRESUMEN
Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.
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Genes del Cloroplasto , Plantas Medicinales/genética , Proteínas Proto-Oncogénicas pp60(c-src)/genética , Rheum/genética , Secuencia de Aminoácidos , Secuencia de Bases , ADN de Plantas/genética , Contaminación de Medicamentos , Genes de Plantas , Genotipo , Datos de Secuencia Molecular , Mutación , Filogenia , Rheum/clasificación , Rizoma/genética , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Ibuprofen/ethyl-cellulose (EC)-polyvinylpyrrolidone (PVP) sustained-release composite particles were prepared by using supercritical CO2 anti-solvent technology. With drug loading as the main evaluation index, orthogonal experimental design was used to optimize the preparation process of EC-PVP/ibuprofen composite particles. The experiments such as encapsulation efficiency, particle size distribution, electron microscope analysis, infrared spectrum (IR), differential scanning calorimetry (DSC) and in vitro dissolution were used to analyze the optimal process combination. The orthogonal experimental optimization process conditions were set as follows: crystallization temperature 40 degrees C, crystallization pressure 12 MPa, PVP concentration 4 mgmL(-1), and CO2 velocity 3.5 Lmin(-1). Under the optimal conditions, the drug loading and encapsulation efficiency of ibuprofen/EC-PVP composite particles were 12.14% and 52.21%, and the average particle size of the particles was 27.621 microm. IR and DSC analysis showed that PVP might complex with EC. The experiments of in vitro dissolution showed that ibuprofen/EC-PVP composite particles had good sustained-release effect. Experiment results showed that, ibuprofen/EC-PVP sustained-release composite particles can be prepared by supercritical CO2 anti-solvent technology.
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Celulosa/análogos & derivados , Composición de Medicamentos , Ibuprofeno/administración & dosificación , Povidona/administración & dosificación , Tecnología Farmacéutica/métodos , Rastreo Diferencial de Calorimetría , Dióxido de Carbono/química , Celulosa/administración & dosificación , Celulosa/química , Cristalización , Preparaciones de Acción Retardada , Portadores de Fármacos , Ibuprofeno/química , Microscopía Confocal , Tamaño de la Partícula , Povidona/química , Solubilidad , Espectrofotometría InfrarrojaRESUMEN
The contents of K, Ca, Na, Mg, Fe, Cu and so on in the roots of Paeonia lactiflora Pall and in the soil in which they grew were determined by ICP-AES technique to study the mineral elements utilization of P. lactiflora Pall. The results indicate that the elements utilization rate of P. lactiflora Pall is different in different locations, so the contents of mineral elements in the roots of P. lactiflora Pall and even the quality of medicinal materials coming from them in different locations are different. The contents of Ca, Fe, Zn and so on in Chi Shao in Heilongjiang and Inner Mongolia are relatively high. The contents of mineral elements in soil influence not only themselves' utilization but also other mineral elements utilization of P. lactiflora Pall.
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Minerales/análisis , Paeonia/química , China , Raíces de Plantas/química , Suelo/químicaRESUMEN
Objective: In Chinese herbal medicine (CHM) history, Lonicerae Japonicae Flos and Lonicerae Flos were used clinically as one drug, but now they are admitted as two herbal medicines in Chinese Pharmacopoeia (2010 edition). This study used network pharmacology to investigate whether the two can be used interchangeably for the treatment of inflammatory diseases in TCM clinical practice. Methods: Lonicerae Japonicae Flos and Lonicerae Flos were compared in the inflammation mechanism including core targets, Gene Ontology (GO), pathway and principle chemical components by the method of network pharmacology. Results: Lonicerae Japonicae Flos and Lonicerae Flos shared in six targets accounting for 66.7% of the entire core targets and more than half of the GO terms and pathways are similar. Organic acids are dominent compounds responsible for anti-inflammatory effects. Three of the compounds that bind to core targets including luteolin, quercetin and kaempferol, are shared in both herbs. Conclusion: Due to high similarity between Lonicerae Japonicae Flos and Lonicerae Flos, we believe that they can be used interchangeably for the inflammation in clinical treatment.
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Affinity analysis is a key biotechnique used in the fields of biology and biomedicine. Herein, we advanced the concept of moving affinity boundary (MAB) using metal ion Ni(II) and histidine (His) as the model inorganic ion and ligand, respectively, developed the simple method of MAB affinity capillary electrophoresis (MAB-ACE), and carried out the relative experiments. The experiments manifested that (a) an MAB could be created with the model metal ion and ligand; (b) the MAB-ACE could specifically capture His rather than other amino acids, or numerous metabolites in human urine; and (c) the capture had the merits of simultaneous focusing and separation to the target metabolite of His. It was further revealed that the specificity of MAB-ACE was originated from the selective affinity interaction and the effective control of affinity conditions. The analyses of His in raw urine by the MAB-ACE are in agreement with those via the standard amino acid analyzer, indicating the reliability of the developed method. Additionally, the MAB-ACE with UV detector had good sensitivity (LOD = 43 ng mL(-1), S/N = 3), 1.0-150 microM linearity and <5% intra-/inter-day variations. The novel method has an evident potential application for capture of a target metabolite in complex biological sample.
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Electroforesis Capilar/métodos , Histidina/aislamiento & purificación , Histidina/orina , Humanos , Níquel/químicaRESUMEN
Soybeans contain oil bodies (OBs) that encapsulate triacylglycerols (TAGs) with a phospholipid monolayer carrying scattered proteins. In nature, soybean OBs can form natural emulsions in aqueous media and may serve as natural, minimally processed, stable, and pre-emulsified oil for addition into appropriate food systems. In this study, OBs were obtained by aqueous extraction from the mature seeds of 2 soybean crop cultivars, high-fat soybean and low-fat soybeans. The compositions of the extracted OBs were analyzed during storage at room temperature up to 14 d (pH = 7). The oxidative stability of these OBs, stored at 60 °C, was evaluated by measuring the presence of primary (lipid hydroperoxides) and secondary lipid oxidation products (malondialdehyde) by determining the standard peroxide value (PV) and thiobarbituric acid-reactive substances (TBARS) value. During storage, the contents of unsaturated fatty acids, phospholipids, and tocopherols declined in both OBs, while their mean particle diameters (d32 ) and ζ-potentials increased. The changes in PV and TBARS values exhibited a similar trend for both OBs, but the OBs from low-fat soybeans had significantly lower PV and higher TBARS values than the OBs from high-fat soybean cultivars (P < 0.05). Overall, the OBs from both soybean cultivars had good stability during storage.
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Glycine max/química , Gotas Lipídicas/química , Aceite de Soja/química , Emulsiones/química , Conservación de Alimentos , Oxidación-Reducción , Semillas/química , Sustancias Reactivas al Ácido TiobarbitúricoRESUMEN
This paper investigated potential utility of capillary zone electrophoresis (CZE) for very succinct but robust quantitative analysis of pyoluteorin (Plt) in anti-fungal fermentation liquor of Pseudomonas species. The experimental conditions for the separation and quantification of Plt were optimized at first. The optimized conditions are: 80 mmol/L pH 8.40 Gly-NaOH buffer, 51 cm total length (42 cm effective) and 75 microm I.D. capillary, 230 nm wavelength, 25 kV, 13 mbar 10s pressure sample injection and 24 degrees C air-cooling. Under the optimized conditions, the migration times of Plt and the internal standard phenobarbital are 2.09 and 2.49 min, respectively, the linear response of Plt concentration ranges from 5.0 to 1000 microg/mL with high correlation coefficient (r=0.99977, n=9), the limits of detection (LOD) and quantification (LOQ) for Plt are 0.66 and 2.2 microg/mL, the precision values (expressed as R.S.D.) of intra- and inter-day are 1.19-1.94% and 1.55-6.21%, respectively, the recoveries of Plt at three concentration levels of 750, 250 and 50 microg/mL range from 90.31% to 97.85% and to 98.96%, respectively. The developed method can be well used for the quantification of Plt in the fermentation liquor.
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Antifúngicos/análisis , Electroforesis Capilar/métodos , Fermentación , Fenoles/análisis , Pseudomonas/metabolismo , Pirroles/análisis , Antifúngicos/aislamiento & purificación , Tampones (Química) , Concentración de Iones de Hidrógeno , Fenoles/aislamiento & purificación , Pirroles/aislamiento & purificación , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
AIM: The aim of this study is to investigate the effect of pre-endurance training on the prevention of alcohol-induced acute hepatic injury and on hepatic mitophagy. METHODS: Forty-eight male Sprague-Dawley rats were randomly divided into four groups: (1) control group, (2) 12-week exercise training group, (3) 5-day alcohol intake group, and (4) 12-week exercise training plus 5-day alcohol intake group. The rats were examined to determine the following: BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3), hypoxia-inducible factor-1α (HIF-1α), cytochrome P450 2E1 (CYP2E1), alcohol dehydrogenase (ADH), microtubule-associated protein 1 light chain 3 (LC3II), Beclin1 mRNA and protein expressions, mitochondrial reactive oxygen species (ROS) production, mitochondrial thiobarbituric acid-reactive substances (TBARS) level, aconitase and ATP synthase activities, mitochondrial inner membrane potential, NADH/NAD(+) ratio, triglyceride (TG), the number of mtDNA and mitochondrial respiration functions in liver tissue, and serum ALT and AST. RESULTS: Pre-endurance training attenuated acute alcohol treatment-induced increase in mitochondrial TBARS, ROS production, NADH/NAD(+) ratio, state 4 respiration rate, TG, serum ALT and AST, as well as BNIP3, HIF-1α, LC3II, and Beclin 1 mRNA and protein levels, however, CYP2E1 and ADH mRNA and protein levels unchanged. Meanwhile, it attenuated the acute alcohol intake-induced decrease in aconitase activity, inner mitochondrial membrane potential (Δψ), ATP synthase activity, state 3 respiration rate, respiratory control ratio, and the number of mtDNA. CONCLUSION: Pre-endurance training can decrease acute alcohol intake-induced damaged mitochondria accumulation and reduced acute alcohol intake-induced mitophagy, which built a new balance between mitophagy and damaged mitochondria accumulation.
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A pot experiment was conducted to study the effect of spent mushroom compost (SMC) in alleviating greenhouse soil secondary salinization and cabbage salt stress. With the amendment of SMC, the salinized soil after 60 day cabbage cultivation had a pH value close to 7.0, its organic matter and available phosphorous contents increased significantly, and the increment of total water-soluble salt content reduced, compared with the control. When the amendment amount of SMC was 10 g x kg(-1), the increment of soil water soluble salt content was the least, suggesting that appropriate amendment with SMC could reduce the salt accumulation in greenhouse soil. Amendment with SMC increased the cabbage seed germination rate, plant height, plant fresh mass, chlorophyll SPAD value, and vitamin C content, and decreased the proline content significantly. All the results indicated that SMC could improve the growth environment of greenhouse cabbage, and effectively alleviate the detrimental effect of salt stress.
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Agricultura/métodos , Brassica/crecimiento & desarrollo , Sales (Química)/análisis , Suelo/análisis , Agaricales , Ambiente Controlado , Eliminación de Residuos/métodos , Estrés FisiológicoRESUMEN
OBJECTIVE: To prepare osteochondral composite scaffold and study its biocompatibility in vitro. METHODS: The composite material of nano-HAP/collagen I was prepared, and the osteochondral scaffold was manufactured by combining nano-HAP, collagen I, and PLGA as the bone section and sodium hyaluronate and PLGA as the chondral section. The diameter, chemical composition and crystallinity of the nano-HAP/collagen I composite particles were assessed with TEM, FTIR and XRD, and the biocompatibility and cytotoxicity of the scaffold were evaluated using MTT assay by co-culturing bone marrow stem cells and the scaffold. RESULTS AND CONCLUSION: The osteochondral composite scaffold has good microstructure without obvious cytotoxicity, possesses good biocompatibility with bone marrow stem cells and is suitable as an osteochondral scaffold material.
Asunto(s)
Materiales Biocompatibles , Durapatita , Ingeniería de Tejidos/métodos , Andamios del Tejido , Células de la Médula Ósea/citología , Células Cultivadas , Condrocitos/citología , Humanos , Ensayo de Materiales , Células Madre Mesenquimatosas/citologíaRESUMEN
An on-line stacking method based on moving reaction boundary (MRB) was developed for the sensitive determination of barbital and phenobarbital in human urine via capillary electrophoresis (CE). The optimized conditions for the method are: 60 mmol L(-1) pH 11.0 Gly-NaOH as the background electrolyte, 10 mmol L(-1) pH 5.5 Gly-HCl as sample buffer, secobarbital as the internal standard (IS), 12.5 kV, 1.4 psi 10s sample injection, 75 microm ID 60.2 cm total length (50 cm effective length) capillary and 214 nm detect wavelength. Under the optimized conditions, the method can well stack and separate barbital and phenobarbital in urine samples and result in 20.5-fold and 22.6-fold improvement in concentration sensitivity for barbital and phenobarbital, respectively. Furthermore, the method holds: (1) good linear calibration functions for the two target compounds (correlation coefficients r>0.999), (2) low limits of detection (0.27 microg mL(-1) for barbital and 0.26 microg mL(-1) for phenobarbital), (3) low limits of quantification (0.92 microg mL(-1) for barbital and 0.87 microg mL(-1) for phenobarbital), (4) good precision (R.S.D. of intra-day and inter-day less than 5.38% for barbital and 1.67% for phenobarbital, respectively) and (5) high recoveries at three concentration levels (90.27-106.36% for barbital and 93.05-113.60% for phenobarbital in urine). The method is simple, sensitive and efficient, and can fit to the need of clinical and forensic toxicology.