RESUMEN
Echinacoside (ECH) is the main compound of Cistanche deserticola, which possesses antioxidant, antitumor, antifatigue, and anti-inflammatory properties. The present study investigated the protective effects of echinacoside on type 2 diabetes mellitus (T2DM)-induced injury in T2DM injury db/db mice model and insulin-resistant LO2 cell model. The results demonstrated that ECH probably alleviated T2DM-induced injury by mediating the AKT pathway, which provided a new direction for the treatment of T2DM-induced injury.
Asunto(s)
Diabetes Mellitus Tipo 2 , Proteínas Proto-Oncogénicas c-akt , Ratones , Animales , Proteínas Proto-Oncogénicas c-akt/metabolismo , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Glicósidos/farmacología , AntioxidantesRESUMEN
Avian influenza viruses (AIV) with good adaptation and various mutations have threatened both human and animals' health. The H7 subtypes have the potential to cause pandemic threats to human health due to the highly pathogenic characteristics. Therefore, it is quite urgent to develop a novel biosensor for rapid and sensitive detection of H7 subtypes. In this work, a biosensor based on luminescence resonance energy transfer (LRET) from BaGdF5:Yb/Er upconversion nanoparticles (UCNPs) to gold nanoparticles (AuNPs) has been developed for rapid and sensitive H7 subtypes detection. The amino modified capture oligonucleotide probes are covalently linked to poly(ethylenimine) (PEI) modified BaGdF5:Yb/Er UCNPs. The thiol modified oligonucleotides with H7 hemagglutinin gene sequence are conjugated to surfaces of AuNPs. The hybridization process between complementary strands of H7 Hemagglutinin gene and its probe brings the energy donor and acceptor into close proximity, leading to the quenching of fluorescence of UCNPs. A linear response is obtained ranging from 10 pm to 10 nm and the limit of detection (LOD) is around 7 pm with detection time around 2 hours. This biosensor is expected to be a valuable diagnostic tool for rapid and sensitive detection of AIV.
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Técnicas Biosensibles/métodos , Transferencia Resonante de Energía de Fluorescencia/métodos , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Virus de la Influenza A/aislamiento & purificación , Gripe Aviar/diagnóstico , Animales , Técnicas Biosensibles/instrumentación , Aves , Transferencia Resonante de Energía de Fluorescencia/instrumentación , Oro , Humanos , Subtipo H7N1 del Virus de la Influenza A/genética , Subtipo H7N1 del Virus de la Influenza A/aislamiento & purificación , Subtipo H7N2 del Virus de la Influenza A/genética , Subtipo H7N2 del Virus de la Influenza A/aislamiento & purificación , Subtipo H7N3 del Virus de la Influenza A/genética , Subtipo H7N3 del Virus de la Influenza A/aislamiento & purificación , Subtipo H7N7 del Virus de la Influenza A/genética , Subtipo H7N7 del Virus de la Influenza A/aislamiento & purificación , Subtipo H7N9 del Virus de la Influenza A/genética , Subtipo H7N9 del Virus de la Influenza A/aislamiento & purificación , Virus de la Influenza A/clasificación , Virus de la Influenza A/genética , Gripe Aviar/virología , Gripe Humana/diagnóstico , Gripe Humana/genética , Límite de Detección , Luminiscencia , Nanopartículas del Metal/química , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
OBJECTIVE: To study the effect of FAM172A protein related to diabetic macroangiopathy on apoptosis and proliferation in HEK293 cells. METHODS: The pDrive-FAM172A plasmid constructed in our previous study was used as a template to amplify human FAM172A open reading frame by a polymerase chain reaction. The resulting PCR products were subcloned into the eukaryotic expression vector PDC315 to construct recombinant PDC315-FAM172A plasmid. PDC315-FAM172A plasmid was identified by enzyme cleavage and sequencing analysis. HEK293 cells were transiently transfected respectively with appropriate PDC315 or PDC315-FAM172A plasmid by Lipofectamine 2000 according to the manufacturer's instruction. XTT assay and growth curve were used to observe the effect of over-expression of FAM172A gene on HEK293 cell proliferation. PI and Annexin V/PI staining method were used to assess the effect of FAM172A gene on apoptosis and cell cycle of HEK293 cell. RESULTS: Eukaryotic expression vector PDC315-FAM172A was successfully constructed and identified by enzyme cleavage and sequencing analysis. Compared with PDC315 plasmid transfection, the XTT assay showed that optical density (A) value increased by 52% when transfected with PDC315-FAM172A plasmid (0.21±0.07 vs 0.32±0.06, P<0.01). Growth curve revealed that HEK293 cells transfected with PDC315-FAM172A plasmid proliferated faster than those transfected with PDC315 plasmid. PI staining showed that, as compared with PDC315 plasmid transfection, the apoptotic rate of HEK293 cells transfected with PDC315-FAM172A plasmid decreased by 38.5% (23.79±1.36 vs 14.64±0.95, P<0.01), cell percentage of G0-G1 phases significantly decreased (66.79±1.73 vs 58.16±0.75, P<0.01) and cell percentage of S phases significantly increased (22.62±1.16 vs 33.56±0.94, P<0.01). Annexin V/PI staining revealed that, as compared with PDC315 plasmid transfection, the percentage of early and advanced apoptotic cells decreased by 28% (13.63±0.56 vs 9.79±0.39, P<0.01) and 29% (7.70±0.29 vs 5.43±0.29, P<0.01) respectively. CONCLUSION: FAM172A protein promotes cell proliferation, inhibits cell apoptosis and facilitates S-phases entry. It indicates that FAM172A protein is involved in cell growth regulation. Our findings provide a clue for further study on its physiological functions and roles in diabetic macroangiopathy.
Asunto(s)
Apoptosis , Proliferación Celular , Proteínas/farmacología , Transfección , Ciclo Celular , Vectores Genéticos , Células HEK293 , Humanos , Riñón/citología , Riñón/embriología , Plásmidos , Proteínas/genéticaRESUMEN
The gut microbiota has been shown to play an important role in chronic liver disease. It has been found that both Lactobacillus rhamnosus and its culture supernatant have the potential to mitigate alcoholic steatohepatitis. However, the exact mechanism is still not fully understood. Bone marrow mesenchymal stem cells have immunosuppressive effects with few side effects. The synergistic effect between Lactobacillus rhamnosus culture supernatant and bone marrow mesenchymal stem cells (BMMSCs) deserves further observation. In this study, a mouse model of chronic alcoholic hepatitis was established by eight weeks of Lieber-DeCarli liquid diet feeding; and LGG-s, BMMSCs or a combination of the two were used to explore a new therapeutic method for alcoholic liver disease and to study the mechanism. The results showed that the combined LGG-s and BMMSC treatment might have a synergistic effect and could improve the symptoms of alcoholic hepatitis by regulating inflammation, autophagy and lymphocyte subsets through the PI3k/NF-kB and PI3K/mTOR pathways. With the treatment, the autophagy rate accelerated, and alcohol-induced natural killer B (NKB) cell and follicular helper T (TFH) cell numbers decreased. These findings suggest that the development of alcoholic hepatitis may occur via PI3K/NF-kB and PI3K/mTOR pathway overactivation as well as through NKB and TFH cell imbalances. Moreover, LGG-s and BMMSCs can regulate these factors and alleviate the disease.
RESUMEN
Angiopoietin-like protein 4 (Angptl4) is a secreted protein predominantly expressed in liver and adipose tissues, and has been identified as an adipokine. Angptl4 is the target gene of peroxisome proliferatoractivated receptors, which are widely used as lipidlowering and antidiabetic drugs, and previous studies have demonstrated that Angptl4 is able to directly stimulate adipocyte lipolysis. The current study focused on how Angptl4 was involved in regulating lipid and glucose metabolism in highfatdiet (HFD) C57 mice. In the present study, mice were divided into three groups, with standard chow mice as a normal control, adenovirus (adv)injected HFD mice as a model control and advAngptl4injected HFD mice as the Angptl4+ group. Firstly, compared with the normal control group, mice in the model control group gained more body weight with severe liver steatosis and increased serum levels of triglyceride, total cholesterol, free fatty acids, alanine aminotransferase and aspartate aminotransferase. In the Angptl4+ group, Angptl4 reduced the weight growth rate, aggravated hepatic steatosis and further increased all the aforementioned serum indexes. Secondly, compared with the normal control, the model control group had a reduced glucose tolerance and developed insulin resistance. Angptl4 expression and the phosphorylation levels of several insulin signaling pathwayassociated genes, insulin receptor substrate 1, protein kinase B, janus kinase 2, signal transducer and activator of transcription 3 were downregulated in the liver samples. AdvAngptl4 injection was observed to improve glucose tolerance and insulin resistance. The genes measured were identified to be upregulated close to normal levels. All the results suggested that Angptl4 served an important role in lipid and glucose metabolism in HFDinduced obese mice, and this may have a great significance for treatment of hyperlipidemia, diabetes, metabolic syndrome and other diseases.
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Angiopoyetinas/metabolismo , Dieta Alta en Grasa/efectos adversos , Hígado Graso/etiología , Hígado Graso/metabolismo , Resistencia a la Insulina , Hígado/metabolismo , Alanina Transaminasa/sangre , Proteína 4 Similar a la Angiopoyetina , Angiopoyetinas/genética , Animales , Aspartato Aminotransferasas/sangre , Colesterol/sangre , Ácidos Grasos/sangre , Hígado Graso/sangre , Hígado Graso/patología , Glucosa/metabolismo , Insulina/metabolismo , Hígado/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Transducción de Señal , Triglicéridos/sangre , Regulación hacia ArribaRESUMEN
OBJECTIVE: The aim of this study was to investigate the prevalence of Mutans streptococci (MS) in children of 3-4 years and thus reveal the relationship between the children's acquisition of MS and their mothers' pathogen. METHODS: Fifty mother-child pairs were selected, examined and divided into three groups according to the children's caries. MS in plaque and mothers' salivary samples were detected by MSB medium. Then 200 MS strains from 20 mothers-children were analyzed by AP-PCR. RESULTS: Acquisition of MS was identified in 37 of 50 children (74%), including 11 of 24 caries-free children and all 26 children with caries. The difference was significant (P<0.01). Genotypes showed that 16 of 37 children (43.2%) had the same fingerprint as their mothers'. The level of MS identified in mothers' salivary sample was lower than that in mothers' plaque sample (32% and 56%). CONCLUSION: These results suggested that caries in children of 3-4 years are closely related with MS acquisition. Mothers are still their important source of MS. The sensitivity of mothers' salivery samples is much lower than that of plaque samples in studying the transmission of MS.
Asunto(s)
Caries Dental/microbiología , Transmisión Vertical de Enfermedad Infecciosa , Madres , Streptococcus mutans/aislamiento & purificación , Adulto , Preescolar , Dermatoglifia del ADN , ADN Bacteriano/genética , Caries Dental/prevención & control , Pruebas de Actividad de Caries Dental , Femenino , Humanos , MasculinoRESUMEN
In this paper, biofunctional magnetic beads were investigated for bacterial cells concentration in a nanoporous alumina membrane based immunosensor for ultra-sensitive detection of E. coli O157:H7. The specific antibody modified magnetic beads were used for concentration of E coli O157:H7 from samples in a small region to enhance sensitivity. The magnetic bead conjugated E. coli O157:H7 cells were then captured on the nanoporous alumina membrane with immobilized antibody via assembled PEG-silane linker. Scanning electron microscopy and fluorescent microscopy were used to demonstrate the magnetic bead-bacteria cell conjugation and bacteria cells magnetic concentration, respectively. Impedance spectroscopy was used to monitor the pure E coli O157:H7 cells and magnetic bead conjugated E coli O157:H7 cells binding on antibody immobilized nanoporous membrane with or without magnetic field. Compared with direct detection of pure bacteria cells, this method via magnetic bead conjugation and concentration demonstrated the ultrasensitivity of 10 CFU/mL for E coli O157:H7 detection.
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Técnicas Biosensibles/instrumentación , Conductometría/instrumentación , Escherichia coli O157/aislamiento & purificación , Inmunoensayo/instrumentación , Separación Inmunomagnética/instrumentación , Membranas Artificiales , Nanoestructuras/química , Anticuerpos Antibacterianos/química , Anticuerpos Antibacterianos/inmunología , Diseño de Equipo , Análisis de Falla de Equipo , Escherichia coli O157/inmunología , Nanoestructuras/ultraestructura , Porosidad , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The objective of this study was to evaluate the influence of a genetic variant in the multidrug resistance 1 gene (MDR1) on hepatocellular carcinoma (HCC) risk. This case-control study was conducted in a Chinese population of 645 HCC cases and 658 cancer-free controls. The genotype of the c.3751G>A genetic variant in the MDR1 gene was investigated by created restriction site-polymerase chain reaction (CRS-PCR) and DNA sequencing methods. Our data demonstrated significantly differences detected in the allelic and genotypic frequencies between HCC cases and those of cancer-free controls. Association analyses indicated that there were statistically increased risk of HCC in the homozygote comparison (AA versus (vs.) GG: OR = 2.22, 95% CI 1.51-3.27, χ(2) = 16.90, P < 0.001), dominant model (AA/GA vs. GG: OR = 1.25, 95% CI 1.00-1.55, χ(2) = 3.98, P = 0.046), recessive model (AA vs. GA/GG: OR = 2.14, 95% CI 1.47-3.09, χ(2) = 16.68, P < 0.001) and allele comparison (A vs. G: OR = 1.33, 95% CI 1.13-1.57, χ(2) = 11.66, P = 0.001). The allele-A and genotype-AA may contribute to HCC susceptibility. These preliminary findings suggest that the c.3751G>A genetic variant in the MDR1 gene is potentially related to HCC susceptibility in a Chinese Han population, and might be used as a molecular marker for evaluating HCC susceptibility.
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Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Pueblo Asiatico/genética , Carcinoma Hepatocelular/etiología , Neoplasias Hepáticas/etiología , Polimorfismo de Nucleótido Simple/genética , Subfamilia B de Transportador de Casetes de Unión a ATP , Carcinoma Hepatocelular/epidemiología , Estudios de Casos y Controles , China/epidemiología , Femenino , Estudios de Seguimiento , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Neoplasias Hepáticas/epidemiología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Pronóstico , Factores de RiesgoRESUMEN
Neurochip based on light-addressable potentiometric sensor (LAPS), whose sensing elements are excitable cells, can monitor electrophysiological properties of cultured neuron networks with cellular signals well analyzed. Here we report a kind of neurochip with rat pheochromocytoma (PC12) cells hybrid with LAPS and a method of de-noising signals based on wavelet transform. Cells were cultured on LAPS for several days to form networks, and we then used LAPS system to detect the extracellular potentials with signals de-noised according to decomposition in the time-frequency space. The signal was decomposed into various scales, and coefficients were processed based on the properties of each layer. At last, signal was reconstructed based on the new coefficients. The results show that after de-noising, baseline drift is removed and signal-to-noise ratio is increased. It suggests that the neurochip of PC12 cells coupled to LAPS is stable and suitable for long-term and non-invasive measurement of cell electrophysiological properties with wavelet transform, taking advantage of its time-frequency localization analysis to reduce noise.