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Histone acetylation plays critical roles in chromatin remodeling, DNA repair, and epigenetic regulation of gene expression, but the underlying mechanisms are unclear. Proteasomes usually catalyze ATP- and polyubiquitin-dependent proteolysis. Here, we show that the proteasomes containing the activator PA200 catalyze the polyubiquitin-independent degradation of histones. Most proteasomes in mammalian testes ("spermatoproteasomes") contain a spermatid/sperm-specific α subunit α4 s/PSMA8 and/or the catalytic ß subunits of immunoproteasomes in addition to PA200. Deletion of PA200 in mice abolishes acetylation-dependent degradation of somatic core histones during DNA double-strand breaks and delays core histone disappearance in elongated spermatids. Purified PA200 greatly promotes ATP-independent proteasomal degradation of the acetylated core histones, but not polyubiquitinated proteins. Furthermore, acetylation on histones is required for their binding to the bromodomain-like regions in PA200 and its yeast ortholog, Blm10. Thus, PA200/Blm10 specifically targets the core histones for acetylation-mediated degradation by proteasomes, providing mechanisms by which acetylation regulates histone degradation, DNA repair, and spermatogenesis.
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Reparación del ADN , Histonas/metabolismo , Proteínas Nucleares/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Espermatogénesis , Testículo/metabolismo , Acetilación , Secuencia de Aminoácidos , Animales , Roturas del ADN de Doble Cadena , Humanos , Masculino , Ratones , Datos de Secuencia Molecular , Proteínas Nucleares/química , Estructura Terciaria de Proteína , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Alineación de SecuenciaRESUMEN
Valproic acid (VPA) is a first-line antiepileptic drug with broad efficacy. Due to significant individual differences in its metabolism, therapeutic drug monitoring is commonly used. However, the recommended therapeutic range (50-100 µg/mL) is inadequate for predicting clinical outcomes. Additionally, the relationship between VPA metabolites and clinical outcomes remains unclear. In this retrospective study, 485 Chinese Southern Han epilepsy patients receiving VPA monotherapy were analyzed after reaching steady-state levels. Plasma concentrations of VPA and its five main metabolites were determined by liquid chromatography-mass spectrometry (LC-MS). We assessed the relevance of the recommended therapeutic VPA range for clinical outcomes and explored the association between VPA/metabolites levels and treatment efficacy/adverse effects. Vitro experiments were conducted to assess 4-ene-VPA hepatotoxicity. The therapeutic range of VPA exhibited no significant correlation with clinical outcomes, and plasma concentrations of VPA failed to serve as predictive indicators for treatment response/adverse effects. Treatment responders had higher 2-PGA concentrations (median, 26.39 ng/mL versus 13.68 ng/mL), with a threshold of 36.5 ng/mL for optimal epilepsy treatment. Patients with abnormal liver function had a higher 4-ene-VPA median concentration (6.41 µg/mL versus 4.83 µg/mL), and the ratio of 4-ene-VPA to VPA better predicted VPA-induced hepatotoxicity (area under the curve, 0.718) than 4-ene-VPA concentration. Vitro experiments revealed that 4-ene-VPA was more hepatotoxic than VPA in HepaRG and L02 cell lines. Total plasma VPA concentration does not serve as a predictor of clinical outcomes. 2-PGA concentrations may be associated with efficacy, whereas the ratio of 4-ene-VPA to VPA may be considered a better biomarker (threshold 10.03%) for VPA-induced hepatotoxicity. SIGNIFICANCE STATEMENT: This was the first and largest observational cohort in China to explore the relationship between patients' parent and metabolites concentrations of VPA and clinical outcomes during the maintenance of VPA monotherapy in epileptic patients. This study provided feasible references of VPA for epilepsy clinical treatment with a larger sample of patients compared with previous studies for a more definitive conclusion based on real-world situations. We found two potential biomarkers in predicting efficacy and liver injury, respectively. This breakthrough has the potential to assist in the rational use of VPA.
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Enfermedad Hepática Inducida por Sustancias y Drogas , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Epilepsia , Humanos , Anticonvulsivantes/efectos adversos , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Monitoreo de Drogas , Epilepsia/tratamiento farmacológico , Estudios Retrospectivos , Ácido Valproico/efectos adversosRESUMEN
The treatment of immunomodulation in multiple sclerosis (MS) can alleviate the severity and relapses. However, it cannot improve the neurological disability of patients due to a lack of myelin protection and regeneration. Therefore, remyelinating therapies may be one of the feasible strategies that can prevent axonal degeneration and restore neurological disability. Natural product icariin (ICA) is a flavonol compound extracted from epimedium flavonoids, which has neuroprotective effects in several models of neurological diseases. Here, we attempt to explore whether ICA has the potential to treat demyelination and its possible mechanisms of action using lipopolysaccharide-treated BV2 microglia, primary microglia, bone marrow-derived macrophages, and cuprizone-induced demyelination model. The indicators of oxidative stress and inflammatory response were evaluated using commercial kits. The results showed that ICA significantly reduced the levels of oxidative intermediates nitric oxide, hydrogen peroxide, malondialdehyde, and inflammatory cytokines TNF-α, IL-1ß, and increased the levels of antioxidants superoxide dismutase, catalase, glutathione peroxidase, and anti-inflammatory cytokines IL-10 and TGF-ß in vitro cell experiments. In vivo demyelination model, ICA significantly alleviated the behavioral abnormalities and enhanced the integrated optical density/mm2 of Black Gold II and myelin basic protein myelin staining, accompanied by the inhibition of oxidative stress/inflammatory response. Immunohistochemical staining showed that ICA significantly induced the expression of nuclear factor erythroid derived 2/heme oxygenase-1 (Nrf2/HO-1) and inhibited the expression of toll-like receptor 4/ nuclear factor kappa B (TLR4/NF-κB), which are two key signaling pathways in antioxidant and anti-inflammatory processes. Our results strongly suggest that ICA may be used as a potential agent to treat demyelination via regulating Nrf2/HO-1-mediated antioxidative stress and TLR4/NF-κB-mediated inflammatory responses.
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Antioxidantes , Enfermedades Desmielinizantes , Flavonoides , Humanos , Antioxidantes/farmacología , Cuprizona/farmacología , Receptor Toll-Like 4 , FN-kappa B , Factor 2 Relacionado con NF-E2 , Antiinflamatorios/farmacología , Citocinas , Enfermedades Desmielinizantes/inducido químicamente , Enfermedades Desmielinizantes/tratamiento farmacológicoRESUMEN
The magneto-optical resonance response of sodium atoms generated by a high-energy solid-state pulse Nd:YAG laser is studied in different external magnetic fields. We investigate the resonance fluorescence signal of sodium atoms in a simulated sea fog environment based on the laser-induced plasma (LIP) effect. By ionizing an NaCl solution spray to generate sodium atoms in an atmospheric environment, we build a Bell-Bloom magneto-optical resonance system under laboratory conditions. With the help of laser-induced breakdown spectroscopy (LIBS) and extinction spectrum, we obtain sodium atoms with a lifetime of 250 µs. A narrowband tunable continuous wave (CW) 589-nm laser tuned at the D2 line with a modulation frequency around the Larmor frequency is used as the pump beam to polarize sodium atoms in the test magnetic field. We find that the magneto-optical resonance signals vary with different external magnetic fields and the positions of the resonance signal are consistent with the theoretical values. An intrinsic magnetometric sensitivity of 620.4 pT in a 1-Hz bandwidth is achieved.
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Twin-field quantum key distribution (TF-QKD) has attracted considerable attention and developed rapidly due to its ability to surpass the fundamental rate-distance limit of QKD. However, the device imperfections may compromise its practical implementations. The goal of this paper is to make it robust against the state preparation flaws (SPFs) and side channels at the light source. We adopt the sending or not-sending (SNS) TF-QKD protocol to accommodate the SPFs and multiple optical modes in the emitted states. We analyze that the flaws of the phase modulation can be overcome by regarding the deviation of the phase as phase noise and eliminating it with the post-selection of phase. To overcome the side channels, we extend the generalized loss-tolerant (GLT) method to the four-intensity decoy-state SNS protocol. Remarkably, by decomposing of the two-mode single-photon states, the phase error rate can be estimated with only four parameters. The practical security of the SNS protocol with flawed and leaky source can be guaranteed. Our results might constitute a crucial step towards guaranteeing the practical implementation of the SNS protocol.
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Traditional Chinese medicine (TCM) has evolved over several thousands of years, which has been shown to be efficacious in the treatment of ischemic heart disease. Three classical TCM prescriptions, namely Xuefu Zhuyu Decoction, Zhishi Xiebai Guizhi Decoction, and Gualou Xiebai Banxia Decoction, have been extensively used in the treatment of coronary heart disease (CHD). Based on molecular network modeling, we performed a comparative pharmacogenomic analysis to systematically determine the drug-targeting spectrum of the three prescriptions at molecular level. Wide-area target molecules of CHD were covered, which was a common feature of the three decoctions, demonstrating their therapeutic functions. Meanwhile, collective signaling involved metabolic/pro-metabolic pathways, driving and transferring pathways, neuropsychiatric pathways, and exocrine or endocrine pathways. These organized pharmacological disturbance was mainly focused on almost all stages of CHD intervention, such as anti-atherosclerosis, lipid metabolism, inflammation, vascular wall function, foam cells formation, platelets aggregation, thrombosis, arrhythmia, and ischemia-reperfusion injury. In addition, heterogeneity analysis of the global pharmacological molecular spectrum revealed that signaling crosstalk, cascade convergence, and key targets were tendentious among the three decoctions. After all, it is unadvisable to rank the findings on targeting advantages of the three decoctions. Comparative pharmacological evidence may provide an appropriate decoction scheme for individualized intervention of CHD.
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Enfermedad Coronaria/tratamiento farmacológico , Enfermedad Coronaria/genética , Medicamentos Herbarios Chinos/uso terapéutico , Pruebas de Farmacogenómica , Humanos , Medicina Tradicional China , Modelos MolecularesRESUMEN
Detecting pesticide residues in human serum is a challenging process. In this study we developed and validated a method for the extraction and recovery of residues of multiple classes of pesticides from serum using one reagent. Salt-assisted acetonitrile extraction and high-performance liquid chromatography with quadrupole time of flight tandem mass spectrometry were used to quantitate 34 pesticides classified in nine groups of chemicals in human serum samples, which are frequently detected in food. The recoveries for 33 of analyzed pesticides ranged from 86 to 112% with relative standard deviations below 15%. The limits of quantitation and linearity of 31 of the pesticides were 1 µg/L and >0.990, respectively. The lower limit of quantitation has been reported in the literature particularly for multi-classes pesticide mixtures in human serum. The salt-acetonitrile reagent was allowed to achieve good recoveries and detection limits, which could be attributed to salt altering the solvent polarity, preferentially collecting the organic phase in the solution, and promoting the extraction. The developed method was applied for two organophosphate pesticide metabolites, diethylphosphate and 3,5,6-trichloro-2-pyridinol, in serum from rats that were fed a nonlethal quantity of chlorpyrifos. The concentrations of these two were 252.18 ± 15.47 and 0.63 ± 0.23 µg/L, respectively.
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Acetonitrilos/química , Compuestos Organofosforados/sangre , Residuos de Plaguicidas/sangre , Plaguicidas/sangre , Cromatografía Líquida de Alta Presión , Humanos , Sales (Química)/química , Espectrometría de Masas en TándemRESUMEN
Danshen (Salvia miltiorrhiza) preparations such as Danhong injection, Danshen injection, Salvianolate injection, compound Danshen injection and Sodium Tanshinone IIA Sulfonate (STS) injection are widely used in China to treat stable angina (angina pectoris) caused by coronary heart disease. In this study we compared the network pharmacological mechanisms of the 5 Danshen preparations. Following a literature search performed in PubMed, EMBASE, Cochrane Library, China National Knowledge Infrastructure (CNKI) database, China Biology Medicine (CBM) database, China Conference Paper Database, Wanfang Database, VIP Database and Conference Proceedings Citation Index (through January 2015), 444 randomized controlled trial publications detailing the use of the 5 Danshen-based injections for treating stable angina were identified, and their combined data were analyzed using a network meta-analysis. All of the 5 Danshen-based preparations were effective in treating stable angina with clinical improvement rates of 72.4%-91.6% and electrocardiogram (ECG) improvement rates of 54.5%-71.6%. According to both clinical improvement and ECG improvement, the 5 Danshen-based preparations were ranked as follows: Danhong injection > Salvianolate injection > STS injection > compound Danshen injection > Danshen injection. There were no significant differences among the safety profiles of the 5 Danshen preparations. The meta-analysis results were further examined using a network pharmacology approach and functional enrichment analysis, which revealed that Danshen and Danhong injections affected 4 and 15 signaling pathways, respectively, and that the 4 signaling pathways affected by Danshen were a subset of those influenced by Danhong. Therefore, Danhong injection affected some unique signaling pathways that might regulate lipoprotein metabolism, oxidation, and inflammation, and protect vascular endothelia, reflecting the multi-component and multi-target characteristics of this traditional formula and its strengths in treating complex diseases.
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Angina Estable/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Medicina Tradicional China/métodos , Salvia miltiorrhiza , Transducción de Señal/efectos de los fármacos , Biología de Sistemas/métodos , Adulto , Anciano , Anciano de 80 o más Años , Angina Estable/diagnóstico , Angina Estable/metabolismo , Angina Estable/fisiopatología , Medicamentos Herbarios Chinos/efectos adversos , Electrocardiografía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Recuperación de la Función , Resultado del TratamientoRESUMEN
Red wines produced in the Xinjiang region of China possess poor color density, and lack fruity notes and elegance. The freeze concentration technique, as a well-established concentration method for liquid food systems, was applied to the Cabernet Sauvignon (Vitis vinifera L.) wine-making process, aiming to investigate its effect on wine quality improvement. Results showed that the freeze concentration treatment did not significantly alter the physicochemical properties of the wine, except for an increase of glycerol and alcoholic content. This technique increased ester contents, as well as decreasing the amount of volatile acids. Higher alcohol contents were also increased, but within an acceptable content range. All taken into consideration, the freeze concentration treated wine showed better fragrance characters according to sensory evaluation. The non-anthocyanin composition was altered by this application, however, the difference disappeared after the aging process. Fortunately, sensory evaluation showed that the treated wine possessed better mouthfeel properties. Anthocyanin contents were enhanced, and effectively stabilized the fresh wine color attributes, resulting in an improvement in appearance of the treated wine. All results considered, it can be concluded that freeze concentration treatment could be a good choice to improve wine quality.
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Congelación , Fenoles/análisis , Fenoles/química , Vitis/química , Vino/análisis , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Fitoquímicos/análisis , Fitoquímicos/química , Espectrometría de Masas en Tándem , GustoRESUMEN
The objective of this work was to study the correlation between the variation of phenolic compounds and sensory characteristics in white wine during bottle storage and to explore the compounds that affected sensory evolution. Chardonnay (Vitis vinifera L. cv.) dry white wines were bottled under six types of stoppers and stored for 18 months. The composition of phenolic compounds was analyzed, and the sensory attributes of these wines were evaluated by professional panel. Multivariate statistical analysis demonstrated that bottle aging period exhibited a more important effect on phenolic compound evolution than stopper type. Most of the phenolic compounds disappeared after 18 months of bottle storage, whereas the wine sensory attributes were significantly improved after 15-month of bottle aging. No strong correlation existed between the phenolic variation and the dissolved oxygen content. Wine color characteristics developed towards better quality accompanying with the reduction of detectable hydroxycinnamic acid derivatives and flavan-3-ols, while the wine mouth-feel was related mainly to gallic acid and ferulic acid ester. This work provided some references for wine producers to select appropriate storage duration for bottled white wine.
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AIM: Our previous studies have showed that ursodeoxycholic acid (UA) and jasminoidin (JA) effectively reduce cerebral infarct volume in mice. In this study we explored the pure synergistic mechanism of these compounds in treatment of mouse cerebral ischemia, which was defined as synergistic actions specific for phenotype variations after excluding interference from ineffective compounds. METHODS: Mice with focal cerebral ischemia were treated with UA, JA or a combination JA and UA (JU). Concha margaritifera (CM) was taken as ineffective compound. Cerebral infarct volume of the mice was determined, and the hippocampi were taken for microarray analysis. Particular signaling pathways and biological functions were enriched based on differentially expressed genes, and corresponding networks were constructed through Ingenuity Pathway Analysis. RESULTS: In phenotype analysis, UA, JA, and JU significantly reduced the ischemic infarct volume with JU being superior to UA or JA alone, while CM was ineffective. As a result, 4 pathways enriched in CM were excluded. Core pathways in the phenotype-positive groups (UA or JA) were involved in neuronal homeostasis and neuropathology. JU-contributing pathways included all UA-contributing and the majority (71.7%) of JA-contributing pathways, and 10 new core pathways whose effects included inflammatory immunity, apoptosis and nervous system development. The functions of JU group included all functions of JA group, the majority (93.1%) of UA-contributing functions, and 3 new core functions, which focused on physiological system development and function. CONCLUSION: The pure synergism between UA and JA underlies 10 new core pathways and 3 new core functions, which are involved in inflammation, immune responses, apoptosis and nervous system development.
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Isquemia Encefálica/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Medicina Tradicional China/métodos , Biología de Sistemas/métodos , Animales , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/genética , Isquemia Encefálica/metabolismo , Minería de Datos , Bases de Datos Genéticas , Modelos Animales de Enfermedad , Combinación de Medicamentos , Sinergismo Farmacológico , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Genómica , Masculino , Ratones , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , Mapas de Interacción de Proteínas , Transducción de Señal/efectos de los fármacos , Integración de SistemasRESUMEN
OBJECTIVE: To investigate the relationship between clinical indicators of CRAB symptoms and antioxidant enzyme activity in patients with multiple myeloma (MM). METHODS: The activity of catalase (CAT), glutathione peroxidase (GPX), and superoxide dismutase (SOD) in the bone marrow supernatants of 44 patients with MM and 12 patients with non-malignant hematological diseases was detected by colorimetric assay, and then the differences in the activity of antioxidant enzymes between the two groups were compared. Furthermore, the relationship between the activity of antioxidant enzymes in the MM group and the levels of serum calcium, serum creatinine (Scr), hemoglobin (Hb), alkaline phosphatase (ALP) as well as bone lesions were analyzed. RESULTS: The antioxidant enzyme activity was lower in MM patients compared with the control group (P < 0.05). When the concentrations of serum calcium and ALP were higher than the normal levels, Hb was lower than 85 g/L, and there were multiple bone lesions, the activity of CAT, SOD and GPX was significantly declined (P < 0.05); When the concentration of Scr≥177 µmol/L, the activity of GPX was significantly declined (P < 0.05). Regression analyses showed that CAT, SOD and GPX were negatively correlated with serum calcium (r =-0.538, r =-0.456, r =-0.431), Scr (r =-0.342, r =-0.384, r =-0.463), and ALP (r =-0.551, r =-0.572, r =-0.482). CONCLUSION: The activity of antioxidant enzymes, including CAT, SOD and GPX, were decreased in patients with MM and they were negatively correlated with some clinical indicators of CRAB symptoms (such as serum calcium, Scr, and ALP), which suggests that promoting the activity of antioxidant enzymes may be beneficial to treat the CRAB symptoms of the patients with MM.
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Antioxidantes , Mieloma Múltiple , Humanos , Fosfatasa Alcalina/sangre , Fosfatasa Alcalina/metabolismo , Antioxidantes/metabolismo , Médula Ósea , Braquiuros , Calcio/sangre , Calcio/metabolismo , Catalasa/sangre , Catalasa/metabolismo , Creatinina/sangre , Glutatión Peroxidasa/sangre , Glutatión Peroxidasa/metabolismo , Mieloma Múltiple/sangre , Mieloma Múltiple/complicaciones , Mieloma Múltiple/enzimología , Mieloma Múltiple/metabolismo , Superóxido Dismutasa/sangre , Superóxido Dismutasa/metabolismoRESUMEN
Maize-soybean compound intercropping has the potential to increase yield and is being tested for spreading in Huang-Huai-hai Plain. However, the main regulatory regions of this cropping pattern on soil microbial communities have not been clarified. In the present study, the tested samples were collected from three maize root zones of bulk soil, rhizosphere soil, and roots under mono- and intercropping planting modes, respectively. The non-rhizosphere soil chemical properties and enzyme activities were determined, and bacterial communities were characterized using high-throughput sequencing of the 16S rRNA gene V3-V4 region. Compared with monocropping, the maize bulk soil electric conductivity ï¼ECï¼, soil organic matter ï¼SOMï¼, available potassium ï¼AKï¼, available phosphorus ï¼APï¼, total nitrogen ï¼TNï¼, and enzyme activities of intercropping were significantly increased. The α diversities and ß diversity of the bacterial community in rhizosphere soil were significantly different between the two planting modes. There were 11 bacteria genera with significantly higher abundance in the rhizosphere soil of compound planting than that of monoculture, and TN, AP, and catalase were the three most important factors contributing to their distribution. The abundances of 8 genera among the 11 genera mentioned above, unclassified Vicinamibacterales, unclassified Geminicoccaceae, MND1, unclassified Gemmatimonadaceae, Acidibacter, unclassified Vicinamibacteraceae, Sphingomonas, and unclassified Comamonadaceae were significantly positively correlated with TN. As for the bacteria distribution in maize root, AK contributed the most and had a significantly negative correlation with unclassified Rhizobiaceae and unclassified Microscillaceae and a positive correlation with Haliangium. Maize-soybean compound intercropping affected mainly the bacterial community of maize rhizosphere and had an evident effect on soil fertilizer cultivation and microbial diversity regulation, which provides a theoretical basis and practical guidance for rational intercropping to maintain agroecosystem biodiversity.
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Agricultura , Bacterias , Glycine max , Raíces de Plantas , Rizosfera , Microbiología del Suelo , Zea mays , Zea mays/crecimiento & desarrollo , Zea mays/microbiología , Glycine max/crecimiento & desarrollo , Glycine max/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Raíces de Plantas/microbiología , Raíces de Plantas/crecimiento & desarrollo , Agricultura/métodos , ARN Ribosómico 16S/genética , Microbiota , Suelo/química , Producción de Cultivos/métodosRESUMEN
In this paper, we described a palladium/norbornene-catalyzed ortho-C-H phosphormethylation of aryl iodides using XCH2P(O)RR', offering a reliable method for the modular synthesis of polysubstituted α-phosphorylated arenes. Alkenylation, hydrogenation, cyanation, methylation, and arylation were all viable termination steps compatible with the reaction. This method demonstrates excellent functional group tolerance and can be extended to the late-stage modification of bioactive molecules. Furthermore, the synthetic transformations of the products demonstrate the practical utility of this reaction.
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In this study, a novel electrochemiluminescence (ECL) biosensor was developed to detect microRNA-21 (miRNA-21) with high sensitivity by leveraging the combined mechanisms of resonance energy transfer (RET) and surface plasmon coupling (SPC). Initially, the glassy carbon electrode (GCE) were coated with Cu-Zn-In-S quantum dots (CZIS QDs), known for their defect-related emission suitable for ECL sensing. Subsequently, a hairpin DNA H3 with gold nanoparticles (Au NPs) attached at the end was modified over the surface of the quantum dots. The Au NPs could effectively quench the ECL signals of CZIS QDs via RET. Further, a significant amount of report DNA was generated through the action of a 3D DNA walker. When the report DNA opened H3-Au NPs, the hairpin structure experienced a conformational change to a linear shape, increasing the gap between the CZIS QDs and the Au NPs. Consequently, the localized surface plasmon resonance ECL (LSPR-ECL) effect replaced ECL resonance energy transfer (ECL-RET). Moreover, the report DNA was released following the addition of H4-Au NPs, resulting in the formation of Au dimers and a surface plasma-coupled ECL (SPC-ECL) effect that enhanced the ECL intensity to 6.97-fold. The integration of new ECL-RET and SPC-ECL biosensor accurately quantified miRNA-21 concentrations from 10-8 M to 10-16 M with a limit of detection (LOD) of 0.08 fM, as well as successfully applied to validate human serum samples.
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Técnicas Biosensibles , ADN , Técnicas Electroquímicas , Mediciones Luminiscentes , MicroARNs , Puntos Cuánticos , Resonancia por Plasmón de Superficie , MicroARNs/análisis , MicroARNs/sangre , Humanos , Técnicas Electroquímicas/métodos , Técnicas Biosensibles/métodos , ADN/química , Puntos Cuánticos/química , Resonancia por Plasmón de Superficie/métodos , Mediciones Luminiscentes/métodos , Oro/química , Límite de Detección , Transferencia de Energía , Nanopartículas del Metal/químicaRESUMEN
Long-term continuous cropping of facility soils could influence soil properties; however, the differences in soil properties among different continuous cropping years are still not well understood. The objective of this study was to explore the effects of continuous cropping years of tomato on the physical and chemical properties and biological characteristics of facility soil. Conventional analysis, high-throughput sequencing, and other methods were used to examine the soil physicochemical properties, soil microbial community diversity, and enzyme activities in facility soil after continuous tomato cropping for 1-3 years, 5-7 years, and more than 10 years. As the continuous tomato cropping years increased, soil bulk density and pH decreased; soil maximal water holding capacity increased; and organic matter, total nitrogen, and total phosphorus accumulated. As continuous cropping years increased, the total salt and EC value decreased with continuous cropping for 5-7 years and increased from 5-7 years to more than 10 years continuous cropping and showed a trend of secondary soil salinization. There was a significant increase in alkaline phosphatase for 1-3 years to 5-7 years continuous tomato cropping. There were significant differences in fungal community abundance among different cropping years. The Simpson index and Shannon index of fungi showed a trend of increasing first and then decreasing with the extension of continuous cropping years and reached the maximum value at 5 years of continuous cropping. The Chao1 index decreased continuously following the cropping years. As continuous cropping years increased, Streptomyces became the dominant bacteria, and Aspergillus and Pseudaleuria became the dominant fungi. The key factors affected by continuous cropping years were available potassium and available nitrogen based on the redundancy analysis. The results of this study lay the foundation for future research on the influence of continuous cropping years on the health of facility soil.
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Microbiota , Solanum lycopersicum , Suelo/química , Microbiología del Suelo , Rizosfera , Hongos , NitrógenoRESUMEN
Detecting pesticide residues in human serum is a challenging process due to trace-level chronic exposure. Several methods using magnetic adsorbents have been developed for analyzing pesticide residue levels in human serum, but it is still difficult to achieve lower quantitative levels, and the adsorption mechanism for extracting pesticides is unclear. Herein, we propose a feasibility concept of using C18-functionalized magnetic nanoparticles for the adsorption of target pesticides, focusing on the extensively used weakly polar pesticides based on molecular dynamics (MD) simulations. To support this, the facilitated target nanoparticles of Fe3O4@SiO2-C18 were synthesized at a size of 12-13 nm with a magnetic saturation of 40 emu/g. After optimizing and establishing the extraction conditions (1.8 mL C18 modifier, 10 mg sorbents, 3 min adsorption time, 1000 µL ACN for desorption eluent at pH 3.8 and 5 min desorption time), which exhibited recovery = 72.3%-118.3% with RSDs = 0.03-6.57, linearity at 0.01-10 ng/mL with R2 = 0.9561-0.9993, and LODs = 0.01-0.30 ng/mL for the 11 weakly polar pesticides in human serum. Furthermore, the mechanism by which the C18 group selectively extracts weakly polar pesticides was confirmed by binding van der Waals and electrostatic interactions under stable and strong binding energy. The extraction process of efficient adsorption and desorption with C18 functional magnetite nanoparticles suggests a simple method for detecting weakly polar pesticides. The concept may lead to a general approach to analyzing multiple pesticide residues in human serum at trace levels.
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Nanopartículas de Magnetita , Residuos de Plaguicidas , Plaguicidas , Cromatografía Líquida de Alta Presión , Humanos , Simulación de Dinámica Molecular , Residuos de Plaguicidas/análisis , Plaguicidas/análisis , Dióxido de Silicio , Extracción en Fase Sólida , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs) are a heterogeneous group of subpopulations with differentially expressed surface markers. CD146 + MSCs correlate with high therapeutic and secretory potency. However, their therapeutic efficacy and mechanisms in premature ovarian failure (POF) have not been explored. METHODS: The umbilical cord (UC)-derived CD146 +/- MSCs were sorted using magnetic beads. The proliferation of MSCs was assayed by dye670 staining and flow cytometry. A mouse POF model was established by injection of cyclophosphamide and busulfan, followed by treatment with CD146 +/- MSCs. The therapeutic effect of CD146 +/- MSCs was evaluated based on body weight, hormone levels, follicle count and reproductive ability. Differential gene expression was identified by mRNA sequencing and validated by RT-PCR. The lymphocyte percentage was detected by flow cytometry. RESULTS: CD146 +/- MSCs had similar morphology and surface marker expression. However, CD146 + MSCs exhibited a significantly stronger proliferation ability. Gene profiles revealed that CD146 + MSCs had a lower levels of immunoregulatory factor expression. CD146 + MSCs exhibited a stronger ability to inhibit T cell proliferation. CD146 +/- MSCs treatment markedly restored FSH and E2 hormone secretion level, reduced follicular atresia, and increased sinus follicle numbers in a mouse POF model. The recovery function of CD146 + MSCs in a reproductive assay was slightly improved than that of CD146 - MSCs. Ovary mRNA sequencing data indicated that UC-MSCs therapy improved ovarian endocrine locally, which was through PPAR and cholesterol metabolism pathways. The percentages of CD3, CD4, and CD8 lymphocytes were significantly reduced in the POF group compared to the control group. CD146 + MSCs treatment significantly reversed the changes in lymphocyte percentages. Meanwhile, CD146 - MSCs could not improve the decrease in CD4/8 ratio induced by chemotherapy. CONCLUSION: UC-MSCs therapy improved premature ovarian failure significantly. CD146 +/- MSCs both had similar therapeutic effects in repairing reproductive ability. CD146 + MSCs had advantages in modulating immunology and cell proliferation characteristics.
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Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Insuficiencia Ovárica Primaria , Animales , Antígeno CD146/genética , Antígeno CD146/metabolismo , Modelos Animales de Enfermedad , Femenino , Atresia Folicular , Hormonas/metabolismo , Humanos , Células Madre Mesenquimatosas/metabolismo , Ratones , Insuficiencia Ovárica Primaria/metabolismo , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Neospora caninum is an obligate intracellular protozoan that causes neosporosis, N. caninum infection is a major cause of abortion in cattle worldwide. Currently, specific treatment for neosporosis is not available. The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is a cytoplasmic protein complex that plays an important role in host defense against N. caninum infection, but the underlying mechanisms are poorly understood. METHODS: The reactive oxygen species (ROS) inhibitor and the ROS inducer, wild-type (WT) and NLRP3-deficient peritoneal macrophages or mice were used to investigate the role of ROS in NLRP3 inflammasome activation and controlling parasite burdens. ROS production, cell death and cell viability, production of inflammasome-mediated IL-1ß or IL-18, cleavage of caspase-1 and NLRP3 expression, as well as parasite burdens were detected. RESULTS: In vitro, N. caninum induced ROS generation in a dose-dependent manner in peritoneal macrophages. The pretreatment of ROS inhibitor N-acetyl-L-cysteine (NAC) significantly attenuated N. caninum-induced ROS production, LDH release, IL-1ß secretion and NLRP3 expression, whereas N. caninum proliferation was notably increased. In contrary, the ROS inducer pyrogallol (PG) significantly enhanced ROS production and NLRP3 inflammasome activity and decreased the parasite burden in N. caninum-infected peritoneal macrophages. NADPH-dependent ROS-mediated NLRP3 inflammasome activation induced by N. caninum can also be confirmed by using the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI). However, the NAC or DPI pre-treatment or PG treatment did not significantly alter N. caninum-induced inflammasome activities and parasite proliferation in Nlrp3-/- peritoneal macrophages. In vivo, IL-18 releases in serum and parasite burdens in peritoneal exudate cells were significantly increased in PG-treated WT mice after infection with N. caninum; however, IL-18 productions and parasite burdens were not changed in PG-treated Nlrp3-/- mice. Furthermore, PG treatment in WT mice infected with N. caninum significantly decreased the mortality, weight loss and parasite burdens in tissues and histopathological lesions. CONCLUSIONS: Neospora caninum-induced NADPH-dependent ROS generation plays an important role in NLRP3 inflammasome activation and controlling parasites. The ROS inducer PG can control N. caninum infection mainly by promoting NLRP3 inflammasome activation. ROS-mediated NLRP3 inflammasome axis can be a potential therapeutic target for neosporosis.
Asunto(s)
Coccidiosis/veterinaria , Inflamasomas/metabolismo , Macrófagos Peritoneales/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Neospora/inmunología , Especies Reactivas de Oxígeno/metabolismo , Animales , Bovinos/parasitología , Coccidiosis/inmunología , Interacciones Huésped-Parásitos , Inmunidad Innata , Macrófagos Peritoneales/parasitología , Ratones , Cultivo Primario de CélulasRESUMEN
BACKGROUND: To observe the effects of Chinese medicine (CM) Polygonum cuspidatum (PC) on adenosine 5'-monophosphate-activated protein kinase (AMPK), forkhead box O3α (FOXO3α), Toll-like receptor-4 (TLR4), NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and monocyte chemoattractant protein-1 (MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism. METHODS: A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups (n=12 per group). A normal group (n=6) was used as the control. Rats in the normal and model groups were administered distilled water (10 mLâ¢kg-1) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol (23.33 mgâ¢kg-1), and 7.46, 3.73, or 1.87 gâ¢kg-1â¢d-1 PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 mRNA and protein levels in renal tissue or serum. RESULTS: Compared with the normal group, the mRNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6th and 8th weeks (P<0.01 or P<0.05). The mRNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated (P<0.01 or P<0.05). Compared with the model group, at the 6th week, the mRNA transcription levels of AMPK in the high- and medium-dose groups, and protein expression levels of AMPKα1, pAMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and pAMPKα1 in the mediumdose PC group, and pAMPKα1 in the low-dose PC group were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium- and low-dose PC groups were down-regulated (P<0.01 or P<0.05). At the 8th week, the mRNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, pAMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription levels of TLR4 in the medium- and low-dose PC groups, NLRP3 in the high- and low-dose PC groups and MCP-1 in the medium- and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated (P<0.01 or P<0.05). CONCLUSION: PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecular mechanism of PC to improve hyperuricemia-mediated immunoinflflammatory metabolic renal damage.