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Human milk oligosaccharides (HMOs) promote the growth and adhesion of bifidobacteria, thus exerting multiple biological functions on intestinal epithelial cells. Bacterial surface proteins play an important role in bacterial-host intestinal epithelial interactions. In this study, we aim to investigate the effects of surface proteins extracted from Bifidobacterium bifidum DNG6 (B. bifidum DNG6) consuming 2'-fucosyllactose (2'-FL) on Caco-2 cells monolayer barrier injury induced by lipopolysaccharide, compared with lactose (Lac) and galacto-oligosaccharides (GOS). Our results indicated that 2'-FL may promote the surface proteins of B. bifidum DNG6 to improve intestinal barrier injury by positively regulating the NF-κB signaling pathway, reducing inflammation(TNF-α reduced to 50.34%, IL-6 reduced to 22.83%, IL-1ß reduced to 37.91%, and IL-10 increased to 63.47%)and strengthening tight junction (ZO-1 2.39 times, Claudin-1 2.79 times, and Occludin 4.70 times). The findings of this study indicate that 2'-FL can further regulate intestinal barrier damage by promoting the alteration of B. bifidum DNG6 surface protein. The findings of this research will also provide theoretical support for the development of synbiotic formulations.
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At present, computational methods for drug repositioning are mainly based on the whole structures of drugs, which limits the discovery of new functions due to the similarities between local structures of drugs. In this article, we, for the first time, integrated the features of chemical-genomics (substructure-domain) and pharmaco-genomics (domain-indication) based on the assumption that drug-target interactions are mediated by the substructures of drugs and the domains of proteins to identify the relationships between substructure-indication and establish a drug-substructure-indication network for predicting all therapeutic effects of tested drugs through only information on the substructures of drugs. In total, 83 205 drug-indication relationships with different correlation scores were obtained. We used three different verification methods to indicate the accuracy of the method and the reliability of the scoring system. We predicted all indications of olaparib using our method, including the known antitumor effect and unknown antiviral effect verified by literature, and we also discovered the inhibitory mechanism of olaparib toward DNA repair through its specific sub494 (o = C-C: C), as it participates in the low synthesis of the poly subfunction of the apoptosis pathway (hsa04210) by inhibiting the Inositol 1,4,5-trisphosphate receptor(s) (ITPRs) and hydrolyzing poly (ADP ribose) polymerases. ElectroCardioGrams of four drugs (quinidine, amiodarone, milrinone and fosinopril) demonstrated the effect of anti-arrhythmia. Unlike previous studies focusing on the overall structures of drugs, our research has great potential in the search for more therapeutic effects of drugs and in predicting all potential effects and mechanisms of a drug from the local structural similarity.
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Biología Computacional , Bases de Datos Factuales , Interacciones Farmacológicas , Reposicionamiento de Medicamentos , Genómica , Humanos , Proteínas/química , Proteínas/metabolismoRESUMEN
BACKGROUND: The selection of endoscopic treatments for small rectal neuroendocrine tumors is controversial. OBJECTIVE: To retrospectively compare the effectiveness and safety of precut endoscopic mucosal resection (EMR-P) and endoscopic submucosal dissection (ESD) for small rectal neuroendocrine tumors (NETs). METHODS: Data from 98 patients with small rectal NETs who were hospitalized at Shenzhen Second People's Hospital between August 2014 and November 2021 were collected. The en bloc resection rate, pathological complete resection rate, radical resection rate, operation time, adverse event rate and hospital stay were compared between the two groups. RESULTS: The operation time in the EMR-P group was significantly shorter than that in the ESD group. The median hospital stay in the EMR-P group was also significantly shorter than that in the ESD group. There were no significant differences between the two groups in terms of the en bloc resection, complete resection or radical resection rates. There was also no significant difference in the incidence of adverse events between the two groups. The delayed bleeding and delayed perforation rates of the two groups were improved after conservative treatment without surgery. There was no significant difference in the rate of positive vertical margins and horizontal margins between the EMR-P group and the ESD group. No local recurrence or metastasis was found during follow-up. CONCLUSION: EMR-P is an effective and safe endoscopic treatment for rectal NETs with a diameter of less than 10 mm. EMR-P is a significantly shorter procedure and requires a shorter hospital stay than ESD. EMR-P does not increase the cut margin positivity rate.
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Resección Endoscópica de la Mucosa , Tumores Neuroendocrinos , Neoplasias del Recto , Humanos , Resección Endoscópica de la Mucosa/efectos adversos , Resección Endoscópica de la Mucosa/métodos , Tumores Neuroendocrinos/cirugía , Tumores Neuroendocrinos/patología , Estudios Retrospectivos , Resultado del Tratamiento , Neoplasias del Recto/cirugía , Neoplasias del Recto/patología , Márgenes de Escisión , Mucosa Intestinal/patología , Recurrencia Local de Neoplasia/epidemiologíaRESUMEN
Influenza viruses continue evolving and have the ability to cause a global pandemic, so it is very important to elucidate its pathogenesis and find new treatment methods. In recent years, proteomics has made important contributions to describing the dynamic interaction between influenza viruses and their hosts, especially in posttranslational regulation of a variety of key biological processes. Protein posttranslational modifications (PTMs) increase the diversity of functionality of the organismal proteome and affect almost all aspects of pathogen biology, primarily by regulating the structure, function, and localization of the modified proteins. Considerable technical achievements in mass spectrometry-based proteomics have been made in a large number of proteome-wide surveys of PTMs in many different organisms. Herein we specifically focus on the proteomic studies regarding a variety of PTMs that occur in both the influenza viruses, mainly influenza A viruses (IAVs), and their hosts, including phosphorylation, ubiquitination and ubiquitin-like modification, glycosylation, methylation, acetylation, and some types of acylation. Integration of these data sets provides a unique scenery of the global regulation and interplay of different PTMs during the interaction between IAVs and their hosts. Various techniques used to globally profiling these PTMs, mostly MS-based approaches, are discussed regarding their increasing roles in mechanical regulation of interaction between influenza viruses and their hosts.
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Gripe Humana , Proteómica , Acetilación , Humanos , Espectrometría de Masas , Fosforilación , Procesamiento Proteico-Postraduccional , Proteoma/genética , Proteoma/metabolismoRESUMEN
The development of cationic polymers as non-viral gene vectors has been hurdled by their high toxicity, thus degradable and biocompatible polymers are urgently demanded. Herein, five polyesters (B3a-B3e) were synthesized based on the ring-opening copolymerization between α-allyl-δ-valerolactone and δ-valerolactone derivatives decorated with alkyl or alkoxyl chains of different lengths, followed by the modification with 1,5,9-triazacyclododecyl ([12]aneN3) through thiol-ene click reactions. The five polyesters effectively condensed DNA into nanoparticles. Of them, B3a with a shorter alkyl chain and B3d with more positive charged units showed stronger DNA condensing performance and can completely retard the migration of DNA at N/P = 1.6 in the presence of DOPE. B3b/DOPE with a longer alkyl chain exhibited the highest transfection efficiency in HeLa cells with 1.8 times of 25 kDa PEI, while B3d/DOPE with more positive charged units exhibited highest transfection efficiency in A549 cells with 2.3 times of 25 kDa PEI. B3b/DOPE and B3d/DOPE successfully delivered pEGFP into zebrafish, which was superior to 25 kDa PEI (1.5 folds and 1.1 folds, respectively). The cytotoxicity measurements proved that the biocompatibility of these polyesters was better than 25 kDa PEI, due to their degradable property in acid environment. The results indicated that these cationic polyesters can be developed as potential non-viral gene vectors for DNA delivery.
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ADN/genética , Técnicas de Transferencia de Gen , Lactonas/química , Nanopartículas/química , Poliésteres/química , Cationes/química , Cationes/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Vectores Genéticos/química , Humanos , Estructura Molecular , Plásmidos/genética , Poliésteres/farmacología , Polimerizacion , Relación Estructura-ActividadRESUMEN
Two-photon fluorescent Acenaphtho[1,2-b]quinoxaline (ANQ) and the hydrophilic di-(triazole-[12]aneN3) moieties were combined through an alkyl chain (ANQ-A-M) or a ß-hairpin motif with two aromatic γ-amino acid residues (ANQ-H-M) to explore their capabilities for in vitro and in vivo gene delivery and tracing. ANQ-A-M and ANQ-H-M showed the same maximum absorption at 420 nm, and their fluorescent intensities around 650 nm were varied in different solvents and became poor in the protic solvents. Gel electrophoresis assays indicated that both compounds completely retarded the migration of pDNA at 20 µM in the presence of DOPE. However, the DNA condensation with ANQ-H-M was not reversible, and the particle size of the corresponding complexes were larger indicated from the SEM and DLS measurements. In vitro transfections indicated ANQ-A-M/DOPE achieved Luciferase and GFP expressions were to be 7.9- and 5.7-fold of those by Lipo2000 in A549 cells respectively. However, ANQ-H-M showed very poor transfection efficiency in Luciferase expression. With the help of single/two-photon fluorescence imaging it clearly demonstrated that the successful transfection of ANQ-A-M was attributed to its cellular uptake, apparent lysosomal escape, and reversible release of DNA; and the poor transfection of ANQ-H-M was resulted from the aggregation of the DNA complexes which prevented them from the cellular uptake, and also the strong binding ability which is not easy to release DNA. ANQ-A-M/DOPE also exhibited robust gene silencing (83% knockdown of Luciferase) and GFP expression (2.47-fold higher) efficiency compared with Lipo2000 in A549 and zebrafish, respectively. The work demonstrated that the linkage structure between fluorescent and di(triazole-[12]aneN3) played the important role for their gene delivery performance, and that ANQ-A-M represents a vector with the strong transfection efficiency in vitro and in vivo as well as the efficient real time bioimaging properties, which is potential for the development in biomedical research.
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Compuestos de Anilina/química , ADN/genética , Colorantes Fluorescentes/química , Técnicas de Transferencia de Gen , Imagen Óptica , Fotones , Quinoxalinas/química , ARN Interferente Pequeño/genética , Compuestos de Anilina/síntesis química , Colorantes Fluorescentes/síntesis química , Vectores Genéticos/síntesis química , Vectores Genéticos/química , Quinoxalinas/síntesis químicaRESUMEN
Copper-catalyzed azide-alkyne cycloaddition (CuAAC) has been widely used in a variety of scientific research, including dynamic proteomics. The current well-established protocols of CuAAC for proteomics analysis introduce labeling tags (azide- or alkyne-containing reagents) at the protein level, followed by downstream analysis by mass spectrometry. In the present study, a new method for proteomic profiling of nascent proteins relying on highly efficient peptide-based click chemistry is proposed, in which the CuAAC reaction was performed at the peptide level, leading to a significant increase in efficiency of the click conjugation reaction. A remarkable improvement in identification rate for spectrum, distinct peptide, and protein was achieved when proteins to be analyzed were proteolytically cleaved into peptides prior to the click conjugation reaction, which would be beneficial to downstream proteomics analysis, especially for the detection of AHA-tagged proteins in very low amounts.
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Péptidos/química , Proteínas/análisis , Proteómica , Alquinos/química , Azidas/química , Catálisis , Química Clic , Cobre/química , Reacción de Cicloadición , Células HEK293 , HumanosRESUMEN
Ni-rich Li-ion cathode materials promise high energy density, but are limited in power density and cycle life, resulting from their poor dynamic characteristics and quick degradation. On the other hand, capacitor electrode materials promise high power density and long cycle life but limited capacities. A joint energy storage mechanism of these two kinds is performed in the material-compositional level in this paper. A valence coupling between carbon π-electrons and O2- is identified in the as-prepared composite material, using a tracking X-ray photoelectron spectroscopy strategy. Besides delivering capacity simultaneously from its LiNi0.8 Co0.1 Mn0.1 O2 and capacitive carbon components with impressive amount and speed, this material shows robust cycling stability by preventing oxygen emission and phase transformation via the discovered valence coupling effect. Structural evolution of the composite shows a more flattened path compared to that of the pure LiNi0.8 Co0.1 Mn0.1 O2 , revealed by the in situ X-ray diffraction strategy. Without obvious phase transformation and losing active contents in this composite material, long cycling can be achieved.
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Recurrence and adverse events after hepatocellular carcinoma (HCC) treatment occur frequently even treated with the most efficient therapy for HCC, liver transplantation. Therefore, better understanding of HCC progression is required to advance the therapeutic strategy of HCC. This study aims to explore the effect and mechanism of small nucleolar RNA host gene 14 (SNHG14) on HCC cell invasion and migration. SNHG14 and miR-656-3p expression in HCC tissues and cells were examined by qRT-PCR. After co-transfection with sh-SNHG14, miR-656-3p inhibitor, miR-656-3p mimic, si-SIRT5, pcDNA3.1-SIRT5 and corresponding negative controls, HepG2 and MHCC97H cell proliferation, invasion and migration were detected. Then the expression levels of SNHG14, miR-656-3p and SIRT5 were measured by qRT-PCR and Western blot. Luciferases reporter gene assay and RNA pull down identified the relation between SNHG14 and miR-656-3p and between miR-656-3p and SIRT5. SNHG14 was upregulated and miR-656-3p was downregulated in HCC cells. Inhibition of SNHG14 could inhibit HepG2 and MHCC97H cell proliferation, invasion and migration. Upregulation of miR-656-3p or knockdown of SIRT5 significantly suppressed the biological process of HepG2 and MHCC97H cells. SNHG14 directly acted on miR-656-3p and SIRT5 was a target gene of miR-656-3p. miR-656-3p inhibitor or pcDNA3.1-SIRT5 could reverse the inhibition of sh-SNHG14 on cell proliferation, invasion and migration of HCC cells. SNHG14 promotes HCC cell invasion and migration through regulating miR-656-3p/SIRT5 axis.
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Carcinoma Hepatocelular/metabolismo , Movimiento Celular , Neoplasias Hepáticas/metabolismo , MicroARNs/metabolismo , Proteínas de Neoplasias/metabolismo , ARN Largo no Codificante/metabolismo , ARN Neoplásico/metabolismo , Transducción de Señal , Sirtuinas/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , MicroARNs/genética , Invasividad Neoplásica , Proteínas de Neoplasias/genética , ARN Largo no Codificante/genética , ARN Neoplásico/genética , Sirtuinas/genéticaRESUMEN
OBJECTIVES: The aim of this study was to develop a radiomics nomogram by combining the optimized radiomics signatures extracted from 2D and/or 3D CT images and clinical predictors to assess the overall survival of patients with non-small cell lung cancer (NSCLC). METHODS: One training cohort of 239 and two validation datasets of 80 and 52 NSCLC patients were enrolled in this study. Nine hundred seventy-five radiomics features were extracted from each patient's 2D and 3D CT images. Least absolute shrinkage and selection operator (LASSO) regression was used to select features and generate a radiomics signature. Cox hazard survival analysis and Kaplan-Meier were performed in both cohorts. The radiomics nomogram was developed by integrating the optimized radiomics signature and clinical predictors, its calibration and discrimination were evaluated. RESULTS: The radiomics signatures were significantly associated with NSCLC patients' survival time. The signature derived from the combined 2D and 3D features showed a better prognostic performance than those from 2D or 3D alone. Our radiomics nomogram integrated the optimal radiomics signature with clinical predictors showed a significant improvement in the prediction of patients' survival compared with clinical predictors alone in the validation cohort. The calibration curve showed predicted survival time was very close to the actual one. CONCLUSIONS: The radiomics signature from the combined 2D and 3D features further improved the predicted accuracy of survival prognosis for the patients with NSCLC. Combination of the optimal radiomics signature and clinical predictors performed better for individualied survival prognosis estimation in patients with NSCLC. These findings might affect trearment strategies and enable a step forward for precise medicine. KEY POINTS: ⢠We found both 2D and 3D radiomics signature have favorable prognosis, but 3D signature had a better performance. ⢠The radiomics signature generated from the combined 2D and 3D features had a better predictive performance than those from 2D or 3D features. ⢠Integrating the optimal radiomics signature with clinical predictors significantly improved the predictive power in patients' survival compared with clinical TNM staging alone.
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Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Imagenología Tridimensional/métodos , Neoplasias Pulmonares/diagnóstico , Nomogramas , Tomografía Computarizada por Rayos X/métodos , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
Nano drug delivery is a promising domain in biomedical theranostics and has aroused more and more attention in recent years. We report here an amphiphilic polymer TPG1, bearing a H2O2-sensitive benzil and an AIE fluorophore tetraphenylethene (TPE) unit, which is able to self-assemble into spherical nanosized micelles in aqueous solution. Doxorubicin (DOX) can be encapsulated into TPG1 micelles efficiently with the loading capability of up to 59% by weight. The benzil moiety could be cleaved via the Baeyer-Villiger type reaction in the presence of H2O2, leading to the decomposition of TPG1 micelles and release of DOX. In vitro studies indicated that DOX-loaded TPG1 micelles can be internalized by cancer cells, followed by unloading encapsulated DOX under the stimulation of H2O2. The drug release process can be monitored by the AIE fluorescence from the degradation products containing a TPE moiety. MTT assays against HeLa and HepG2 cancer cells demonstrated that DOX-loaded micelles showed good anticancer efficacy. The polymer TPG1 and the corresponding decomposed products showed great biocompatibility. Our data suggest that TPG1 has the potential to be employed for the controlled drug delivery system.
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Doxorrubicina/farmacología , Sistemas de Liberación de Medicamentos , Colorantes Fluorescentes/química , Peróxido de Hidrógeno/farmacología , Fenilglioxal/análogos & derivados , Polímeros/farmacología , Estilbenos/química , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/química , Portadores de Fármacos/química , Portadores de Fármacos/farmacología , Células HeLa , Células Hep G2 , Humanos , Peróxido de Hidrógeno/química , Micelas , Estructura Molecular , Imagen Óptica , Fenilglioxal/química , Fenilglioxal/farmacología , Polímeros/químicaRESUMEN
A 71-year-old man came to our hospital for endoscopic treatment of a suspicious early gastric carcinoma. A 0.6×0.4 cm protrusive esophageal lesion with smooth surface was found accidentally, located at about 20 cm from the incisors. The lesion was successfully resected by endoscopic mucosal resection, which was esophageal cyst containing heterotopic gastric glands.
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Coristoma/patología , Quiste Esofágico/patología , Mucosa Gástrica , Anciano , Coristoma/diagnóstico por imagen , Endosonografía , Quiste Esofágico/diagnóstico por imagen , Humanos , Hallazgos Incidentales , MasculinoRESUMEN
Glycogen synthase kinase-3 beta (GSK-3ß) is involved in multiple signaling pathways. Consistent with its critical roles in normal cells, abnormalities in GSK-3ß activity have been implicated in diabetes, heart disease, Parkinson disease, and Alzheimer's disease. In this study, a series of new scaffolds of small molecule inhibitors of GSK-3ß were identified by virtual screening and bioassay. Candidates that adhere to drug-like criteria from a virtual library of compounds were tested using computational docking studies. Twenty selected compounds were tested, which led to the discovery of two hits. Compound 14 (IC50 = 8.48 µM) and compound 19 (IC50 = 2.19 µM) were identified with high affinity. Molecular dynamics (MD) simulations, in conjunction with molecular mechanics/Poisson-Boltzmann surface area binding free-energy analysis, were employed to gain insight into the binding modes and energetics of GSK-3ß inhibitors. The detailed analysis of molecular dynamics results shows that Ile62, Val70, Tyr134, and Leu188 in GSK-3ß are key residues responsible to the binding of compound 14 and compound 19. Importantly, our results also validated this combined virtual screening and biophysical technique approach to discovery kinase inhibitors, which may be applied for future inhibitor discovery work for GSK-3ß.
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Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Glucógeno Sintasa Quinasa 3 beta/química , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Ensayo de Cambio de Movilidad Electroforética , Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Estructura Secundaria de ProteínaRESUMEN
Cardiac glycosides are natural compounds used for the treatment of congestive heart failure and cardiac arrhythmias. Recently, they have been reported to exhibit anticancer activity. Proscillaridin A (PSN-A), a cardiac glycoside constituent of Urginea maritima has been shown to exhibit anticancer activity. However, the cellular targets and anticancer mechanism of PSN-A in various cancers including prostate cancer remain largely unexplored. In the present study, we have shown that PSN-A inhibits proliferation and induces apoptosis in prostate cancer cells in a dose-dependent manner. Further mechanistic study have shown that anticancer activity of PSN-A in prostate cancer cells is associated with ROS generation, Bcl-2 family proteins modulation, mitochondrial membrane potential disruption and ultimately activation of caspase-3 and cleavage of PARP. Moreover, we found that PSN-A inhibits JAK2/STAT3 signaling and augments doxorubicin toxicity in prostate cancer cells. Of note, LNCaP cells were found to be more sensitive to PSN-A treatment as compared to DU145 cells. Taken together, the data provided first evidence of the anticancer activity and possible molecular mechanism of PSN-A in prostate cancer. Further study is needed to develop PSN-A into a potential lead compound for the treatment of prostate cancer.
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Apoptosis/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Proscilaridina/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Factor de Transcripción STAT3/metabolismo , Antibióticos Antineoplásicos/toxicidad , Línea Celular Tumoral , Proliferación Celular , Doxorrubicina/toxicidad , Humanos , Masculino , Factor de Transcripción STAT3/efectos de los fármacosRESUMEN
There is a significant unmet need in the treatment of primary biliary cirrhosis (PBC) despite significant data on the effector pathways that lead to biliary duct damage. We focused attention on a murine model of PBC, the dominant negative transforming growth factor ß receptor II (Tg) mice. To further define the pathways that lead to biliary pathology in these mice, we developed Tg mice deleted of CD4 cells (CD4(-/-)Tg). Interestingly, these mice developed more severe cholangitis than control Tg mice. These mice, which lack CD4 cells, manifested increased levels of IFN-γ produced by effector CD8 cells. It appears that increased cholangitis is due to the absence of CD4 Treg cells. Based on these data, we parabiosed CD4(-/-)Tg mice with established disease at 8-9 weeks of age with C57BL/6 control mice. Such parabiotic "twins" had a significant reduction in autoimmune cholangitis, even though they had established pathology at the time of surgery. We prepared mixed bone marrow chimera mice constructed from CD4(-/-)Tg and CD8(-/-) mice and not only was cholangitis improved, but a decrease in terminally differentiated CD8(+) T effector cells in the presence of wild type CD4 cells was noted. In conclusion, "correcting" the CD4 T cell subset, even in the presence of pathogenic CD8 T cells, is effective in treating autoimmune cholangitis.
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Enfermedades Autoinmunes/cirugía , Colangitis/cirugía , Cirrosis Hepática Biliar/cirugía , Parabiosis/métodos , Animales , Enfermedades Autoinmunes/inmunología , Conductos Biliares/inmunología , Conductos Biliares/patología , Antígenos CD4/genética , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Colangitis/inmunología , Modelos Animales de Enfermedad , Femenino , Trasplante de Células Madre Hematopoyéticas/métodos , Humanos , Hígado/inmunología , Hígado/patología , Cirrosis Hepática Biliar/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Mutación , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/inmunología , Receptor Tipo II de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/genética , Receptores de Factores de Crecimiento Transformadores beta/inmunologíaRESUMEN
AIM: Tubeimoside-1 (TBMS1), a triterpenoid saponin extracted from the Chinese herbal medicine Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), has shown anticancer activities in various cancer cell lines. The aim of this study was to investigate the anticancer activity and molecular targets of TBMS1 in human prostate cancer cells in vitro. METHODS: DU145 and P3 human prostate cancer cells were treated with TBMS1. Cell viability and apoptosis were detected. ROS generation, mitochondrial membrane potential and cell cycle profile were examined. Western blotting was used to measure the expression of relevant proteins in the cells. RESULTS: TBMS1 (5-100 µmol/L) significantly suppressed the viability of DU145 and P3 cells with IC50 values of approximately 10 and 20 µmol/L, respectively. Furthermore, TBMS1 dose-dependently induced apoptosis and cell cycle arrest at G0/G1 phase in DU145 and P3 cells. In DU145 cells, TBMS1 induced mitochondrial apoptosis, evidenced by ROS generation, mitochondrial dysfunction, endoplasmic reticulum stress, modulated Bcl-2 family protein and cleaved caspase-3, and activated ASK-1 and its downstream targets p38 and JNK. The G0/G1 phase arrest was linked to increased expression of p53 and p21 and decreased expression of cyclin E and cdk2. Co-treatment with Z-VAD-FMK (pan-caspase inhibitor) could attenuate TBMS1-induced apoptosis but did not prevent G0/G1 arrest. Moreover, co-treatment with NAC (ROS scavenger), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor) or salubrinal (ER stress inhibitor) significantly attenuated TBMS1-induced apoptosis. CONCLUSION: TBMS1 induces oxidative stress-mediated apoptosis in DU145 human prostate cancer cells in vitro via the mitochondrial pathway.
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Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Fase G1/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Neoplasias de la Próstata/patología , Fase de Descanso del Ciclo Celular/efectos de los fármacos , Saponinas/farmacología , Triterpenos/farmacología , Acetilcisteína/farmacología , Clorometilcetonas de Aminoácidos/farmacología , Antracenos/farmacología , Caspasa 3 , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cinamatos/farmacología , Ciclina E/metabolismo , Quinasa 2 Dependiente de la Ciclina/metabolismo , Relación Dosis-Respuesta a Droga , Estrés del Retículo Endoplásmico/efectos de los fármacos , Humanos , Imidazoles/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , MAP Quinasa Quinasa Quinasa 5/metabolismo , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Piridinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Saponinas/antagonistas & inhibidores , Tiourea/análogos & derivados , Tiourea/farmacología , Triterpenos/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Thymic CD4(+) FoxP3(+) regulatory T (Treg) cells are critical for the development of immunological tolerance and immune homeostasis and requires contributions of both thymic dendritic and epithelial cells. Although B cells have been reported to be present within the thymus, there has not hitherto been a definition of their role in immune cell development and, in particular, whether or how they contribute to the Treg cellular thymic compartment. Herein, using both phenotypic and functional approaches, we demonstrate that thymic B cells contribute to the maintenance of thymic Treg cells and, using an in vitro culture system, demonstrate that thymic B cells contribute to the size of the thymic Treg compartment via cell-cell MHC II contact and the involvement of two independent co-stimulatory pathways that include interactions between the CD40/CD80/CD86 co-stimulatory molecules. Our data also suggest that thymic B cells promote the generation of thymic Treg cell precursors (pre-Treg cells), but not the conversion of FoxP3(+) Treg cells from pre-Treg cells. In addition, thymic B cells directly promote the proliferation of thymic Treg cells that is MHC II contact dependent with a minimal if any role for co-stimulatory molecules including CD40/CD80/CD86. Both pathways are independent of TGFß. In conclusion, we rigorously define the critical role of thymic B cells in the development of thymic Treg cells from non-Treg to precursor stage and in the proliferation of mature thymic Treg cells.
Asunto(s)
Linfocitos B/inmunología , Proliferación Celular , Linfocitos T Reguladores/inmunología , Timo/inmunología , Animales , Linfocitos B/metabolismo , Antígeno B7-1/inmunología , Antígeno B7-2/inmunología , Antígenos CD40/inmunología , Diferenciación Celular/inmunología , Citometría de Flujo , Antígenos de Histocompatibilidad Clase II/inmunología , Homeostasis/inmunología , Tolerancia Inmunológica/inmunología , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Fluorescente , Linfocitos T Reguladores/metabolismoRESUMEN
CD4(+)Foxp3(+) regulatory T cells (Tregs) play a non-redundant role in control of excessive immune responses, and defects in Tregs have been shown both in patients and murine models of primary biliary cirrhosis (PBC), a progressive autoimmune biliary disease. Herein, we took advantage of a murine model of PBC, the dominant negative transforming growth factor ß receptor II (dnTGFßRII) mice, to assess Treg genetic defects and their functional effects in PBC. By using high-resolution microarrays with verification by PCR and protein expression, we found profound and wide-ranging differences between dnTGFßRII and normal, wild type Tregs. Critical transcription factors were down-regulated including Eos, Ahr, Klf2, Foxp1 in dnTGFßRII Tregs. Functionally, dnTGFßRII Tregs expressed an activated, pro-inflammatory phenotype with upregulation of Ccl5, Granzyme B and IFN-γ. Genetic pathway analysis suggested that the primary effect of loss of TGFß pathway signaling was to down regulate immune regulatory processes, with a secondary upregulation of inflammatory processes. These findings provide new insights into T regulatory genetic defects; aberrations of the identified genes or genetic pathways should be investigated in human PBC Tregs. This approach which takes advantage of biologic pathway analysis illustrates the ability to identify genes/pathways that are affected both independently and dependent on abnormalities in TGFß signaling. Such approaches will become increasingly useful in human autoimmunity.
Asunto(s)
Cirrosis Hepática Biliar/inmunología , Linfocitos T Reguladores/inmunología , Animales , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Modelos Animales de Enfermedad , Femenino , Factores de Transcripción Forkhead/metabolismo , Regulación de la Expresión Génica/genética , Granzimas/metabolismo , Humanos , Mediadores de Inflamación/metabolismo , Interferón gamma/metabolismo , Cirrosis Hepática Biliar/genética , Ratones , Ratones Mutantes , Análisis por Micromatrices , Mutación/genética , Proteínas Serina-Treonina Quinasas/genética , Receptor Tipo II de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/genética , Transducción de Señal , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Avian influenza A viruses are serious veterinary pathogens that normally circulate among avian populations, causing substantial economic impacts. Some strains of avian influenza A viruses, such as H5N1, H9N2, and recently reported H7N9, have been occasionally found to adapt to humans from other species. In order to replicate efficiently in the new host, influenza viruses have to interact with a variety of host factors. In the present study, H7N9 nucleoprotein was transfected into human HEK293T cells, followed by immunoprecipitated and analyzed by proteomics approaches. A series of host proteins co-immunoprecipitated were identified with high confidence, some of which were found to be acetylated at their lysine residues. Bioinformatics analysis revealed that spliceosome might be the most relevant pathway involved in host response to nucleoprotein expression, increasing our emerging knowledge of host proteins that might be involved in influenza virus replication activities.