RESUMEN
OBJECTIVE: To study the effect and mechanism of the Clostridium metabolite p-Cresol sulfate (PCS) in primary biliary cholangitis (PBC). METHODS: Gas chromatography-mass spectrometry (GC-MS) was used to detect differences in tyrosine, phenylalanine, tryptophan, PCS, and p-Cresyl glucuronide (PCG) between the serum of PBC patients and healthy controls. In vivo experiments, mice were divided into the normal control, PBC group, and PBC tyrosine group. GC-MS was used to detect PCS and PCG. Serum and liver inflammatory factors were compared between groups along with the polarization of liver Kupffer cells. Additionally, PCS was cultured with normal bile duct epithelial cells and Kupffer cells, respectively. PCS-stimulated Kupffer cells were co-cultured with lipopolysaccharide-injured bile duct epithelial cells to detect changes in inflammatory factors. RESULTS: Levels of tyrosine and phenylalanine were increased, but PCS level was reduced in PBC patients, with PCG showing a lower concentration distribution in both groups. PCS in PBC mice was also lower than those in normal control mice. After oral administration of tyrosine feed to PBC mice, PCS increased, liver inflammatory factors were decreased, and anti-inflammatory factors were increased. Furthermore, Kupffer cells in the liver polarized form M1 transitioned to M2. PCS can damage normal bile duct epithelial cells and suppress the immune response of Kupffer cells. But PCS protects bile duct epithelial cells damaged by LPS through Kupffer cells. CONCLUSIONS: PCS produced by Clostridium-metabolized tyrosine reduced PBC inflammation, suggesting that intervention by food, or supplementation with PCS might represent an effective clinical strategy for treating PBC.
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Cirrosis Hepática Biliar , Ratones , Animales , Cirrosis Hepática Biliar/tratamiento farmacológico , Cirrosis Hepática Biliar/metabolismo , Macrófagos del Hígado/metabolismo , Sulfatos , Inflamación , Lipopolisacáridos/farmacología , Tirosina , Clostridium , FenilalaninaRESUMEN
A drug delivery system of forsythoside A-loaded exosomes(FTA-Exos) with high biocompatibility and low immunogenicity was established to investigate its impact on the migration of human lung epithelial adenocarcinoma A549 cells. The exosomes from A549 cells were extracted and purified by ultra-high speed centrifugation and ultrafiltration. FTA-Exos were prepared by ultrasonic incubation, and characterized by particle size analysis, transmission electron microscopy, and Western blot assay. The uptake of FTA-Exos by A549 cells was observed under the laser confocal microscope, and the impact of FTA-Exos on the migration of A549 cells was investigated by cell scratch assay. The results showed that the average particle size of the prepared FTA-Exos was(138.90±2.37) nm, which increased slightly after drug loading. The PDI was 0.291±0.013, and the average potential was(-10.1±0.66) mV. The FTA-Exos were spheroidal in appearance as observed by transmission electron microscope, with an obvious saucer-like double-layer membrane. Western blot assay indicated that the specific proteins CD63 and Alix were both expressed in exosomes. The laser confocal microscopy suggested that FTA-Exos were taken up by A549 cells and stably maintained in the cell for 4-8 h, and the fluorescence was significantly enhanced at 4 h. The scratch assay showed that the inhibitory effect of FTA-Exos on the migration of A549 cells was significantly stronger than that of forsythoside A(P < 0.05). In conclusion, the drug delivery system of FTA-Exos established in this study had good stability, reliable preparation process, and potent inhibitory effect on the migration of A549 cells in vitro, which can provide an important reference for subsequent in-depth research and application.
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Exosomas , Glicósidos , HumanosRESUMEN
BACKGROUND Kupffer cells and natural killer (NK) cells has been identified as contributing factors in the pathogenesis of hepatitis, but the detailed mechanism of these cell types in the pathogenesis of primary biliary cholangitis (PBC) is poorly understood. MATERIAL AND METHODS In this study, polyinosinic: polycytidylic acid (poly I: C), 2-octynoic acid-bovine serum albumin (2OA-BSA) and Freund's adjuvant (FA) were injected to establish a murine PBC model, from which NK cells and Kupffer cells were extracted and isolated. The cells were then co-cultivated in a designed culture system, and then NK group 2, member D (NKG2D), retinoic acid early inducible-1 (RAE-1), F4/80, and cytokine expression levels were detected. RESULTS The results showed close crosstalk between Kupffer cells and NK cells. PBC mice showed increased surface RAE-1 protein expression and Kupffer cell cytokine secretion, which subsequently activated NK cell-mediated target cell killing via NKG2D/RAE-1 recognition, and increased inflammation. NK cell-derived interferon-γ (IFN-γ) and Kupffer cell-derived tumor necrosis factor alpha (TNF-alpha) were found to synergistically regulate inflammation. Moreover, interleukin (IL)-12 and IL-10 improved the crosstalk between NK cells and Kupffer cells. CONCLUSIONS Our ï¬ndings in mice are the first to suggest the involvement of the NKG2D/RAE-1 interaction and cytokines in the synergistic effects of NK and Kupffer cells in PBC.
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Células Asesinas Naturales/metabolismo , Macrófagos del Hígado/metabolismo , Cirrosis Hepática Biliar/metabolismo , Animales , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Células Asesinas Naturales/patología , Macrófagos del Hígado/patología , Cirrosis Hepática Biliar/fisiopatología , Ratones , Ratones Endogámicos C57BL , Subfamilia K de Receptores Similares a Lectina de Células NK/metabolismo , Proteínas Asociadas a Matriz Nuclear/metabolismo , Proteínas de Transporte Nucleocitoplasmático/metabolismoRESUMEN
The 59 1st-3rd order tributaries in the middle reach of Qiantang River are negatively affected by different intensities of urbanization. In April 2010, an investigation was conducted on the water bodies' physical and chemical properties and macrobenthos communities of the tributaries, with the relationships between the tributaries' water quality and biological communities and the percentage of ground surface impervious area (PIA), an indicator of urbanization intensity. The Spearman correlation analysis showed that the water bodies' NH(4+)-N, PO4(3-)-P, TP, COD(Mn), conductivity, width, depth, and fine sand/silt ratio were positively correlated with PIA, and negatively correlated with forest land area. The fitted nonlinear regression equations revealed that all the test macro-benthic invertebrate's parameters had significant relationships with PIA, of which, the total number of taxa, Shannon diversity index, richness index, EPT (%), predators (%), shredders (%), filterers (%) and scrapers (%) were negatively correlated to PIA but positively correlated to forest land area, and the BI, collectors (%), tolerance taxa (%) and oligochaeta (%) were positively correlated to the PIA. Our study indicated that under the impact of urbanization, these tributaries presented the common features of degradation, i. e., high concentrations of nitrogen and phosphorus, degradation of physical habitat, disappearance of pollution-sensitive macro-benthic invertebrate species, and dramatic increase of pollution-tolerant species individuals.
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Invertebrados/crecimiento & desarrollo , Nitrógeno/análisis , Fósforo/análisis , Urbanización , Contaminantes Químicos del Agua/análisis , Calidad del Agua , Animales , China , Monitoreo del Ambiente/métodos , Invertebrados/clasificación , Dinámica Poblacional , RíosRESUMEN
Water eutrophication is a global problem, and one of the major environmental problems in China. At present, its studies in China are mainly focused on lakes and reservoirs, whereas a few on rivers. Based on the stress-response of benthic macroinvertebrate assemblages to water nutrients, we used the nonparametric deviance reduction (change point analysis) to compute the breakpoints of nutrients concentrations, in the upper reaches of Xitiao Stream in Zhejiang. The results indicated that the breakpoints of total nitrogen (TN) and total phosphors (TP) were 1.409 mg x L(-1) and 0.033-0.035 mg x L(-1), respectively. The TN and TP concentrations at reference sites were lower, while those at urban sewage- polluted sites were higher than the thresholds. In latter case, a serious degradation of benthic macroinvertebrates assemblages could be induced. To establish water nutrients criteria with relation to aquatic organisms would make the biological monitoring play a full role in water management, and provide scientific data to estimate the total maximum daily load (TMDL) of TN and TP in water body.
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Monitoreo del Ambiente/métodos , Invertebrados/clasificación , Nitrógeno/análisis , Fósforo/análisis , Contaminantes Químicos del Agua/análisis , Animales , China , Invertebrados/crecimiento & desarrollo , Dinámica Poblacional , RíosRESUMEN
Watermelon is a popular fruit in the world with soluble solids content (SSC) being one of the major characteristics used for assessing its quality. This study was aimed at obtaining a method for nondestructive SSC detection of watermelons by means of visible/near infrared (Vis/NIR) diffuse transmittance technique. Vis/NIR transmittance spectra of intact watermelons were acquired using a low-cost commercially available spectrometer operating over the range 350~1000 nm. Spectra data were analyzed by two multivariate calibration techniques: partial least squares (PLS) and principal component regression (PCR) methods. Two experiments were designed for two varieties of watermelons [Qilin (QL), Zaochunhongyu (ZC)], which have different skin thickness range and shape dimensions. The influences of different data preprocessing and spectra treatments were also investigated. Performance of different models was assessed in terms of root mean square errors of calibration (RMSEC), root mean square errors of prediction (RMSEP) and correlation coefficient (r) between the predicted and measured parameter values. Results showed that spectra data preprocessing influenced the performance of the calibration models. The first derivative spectra showed the best results with high correlation coefficient of determination [r=0.918 (QL); r=0.954 (ZC)], low RMSEP [0.65 degrees Brix (QL); 0.58 degrees Brix (ZC)], low RMSEC [0.48 degrees Brix (QL); 0.34 degrees Brix (ZC)] and small difference between the RMSEP and the RMSEC by PLS method. The nondestructive Vis/NIR measurements provided good estimates of SSC index of watermelon, and the predicted values were highly correlated with destructively measured values for SSC. The models based on smoothing spectra (Savitzky-Golay filter smoothing method) did not enhance the performance of calibration models obviously. The results indicated the feasibility of Vis/NIR diffuse transmittance spectral analysis for predicting watermelon SSC in a nondestructive way.
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Algoritmos , Citrullus/química , Análisis de los Alimentos/métodos , Frutas/química , Fotometría/métodos , Extractos Vegetales/análisis , Espectrofotometría Infrarroja/métodos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , SolubilidadRESUMEN
The objective of the present research was to evaluate the potential of NIR diffuse reflectance spectroscopy as a way for nondestructive measurement of loquat soluble solid content of three varieties harvested from two orchards. According to the analysis, it was shown that the correlation coefficient of curves was relatively high in the two regions of 1400-1500 nm and 1900- 2000 nm. Statistical models were developed using partial least square regression (PLSR), stepwise multiple linear regression (SMLR) and principal component regression (PCR) methods in the full spectral region from 800 to 2500 nm and in the combined region of 1400-1500 nm and 1900-2000 nm. The results of PLSR model in the full spectral region were better than those of other models. The modeling results based on derivative spectra were not as good as those based on original spectra. Finally, a model was established based on the original spectra with 17 points smoothing for all the three varieties in the full spectral region by PLSR method. The correlation coefficients of calibration and validation were 0.96 and 0.95, respectively. The results indicate that it is feasible to use NIR spectroscopy technique for quantitative analysis of loquat soluble solid content.