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1.
Mol Pharm ; 21(3): 1300-1308, 2024 Mar 04.
Artículo en Inglés | MEDLINE | ID: mdl-38294949

RESUMEN

Keratin and lipid structures in the stratum corneum (SC) are closely related to the SC barrier function. The application of penetration enhancers (PEs) disrupts the structure of SC, thereby promoting infiltration. To quantify these PE-induced structural changes in SC, we used confocal Raman imaging (CRI) and polarized Raman imaging (PRI) to explore the integrity and continuity of keratin and lipid structures in SC. The results showed that water is the safest PE and that oleic acid (OA), sodium dodecyl sulfate (SDS), and low molecular weight protamine (LMWP) disrupted the ordered structure of keratin, while azone and liposomes had less of an effect on keratin. Azone, OA, and SDS also led to significant changes in lipid structure, while LMWP and liposomes had less of an effect. Establishing this non-invasive and efficient strategy will provide new insights into transdermal drug delivery and skin health management.


Asunto(s)
Liposomas , Piel , Liposomas/farmacología , Epidermis , Ácido Oléico/farmacología , Queratinas
2.
Int J Mol Sci ; 23(4)2022 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-35216331

RESUMEN

A highly efficient Agrobacterium-mediated transformation method is needed for the molecular study of model tree species such as hybrid poplar 84K (Populus alba × P. glandulosa cv. '84K'). In this study, we report a callus-based transformation method that exhibits high efficiency and reproducibility. The optimized callus induction medium (CIM1) induced the development of calli from leaves with high efficiency, and multiple shoots were induced from calli growing on the optimized shoot induction medium (SIM1). Factors affecting the transformation frequency of calli were optimized as follows: Agrobacterium concentration sets at an OD600 of 0.6, Agrobacterium infective suspension with an acetosyringone (AS) concentration of 100 µM, infection time of 15 min, cocultivation duration of 2 days and precultivation duration of 6 days. Using this method, transgenic plants are obtained within approximately 2 months with a transformation frequency greater than 50%. Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and ß-galactosidase (GUS) histochemical staining analyses confirmed the successful generation of stable transformants. Additionally, the calli from leaves were subcultured and used to obtain new explants; the high transformation efficiency was still maintained in subcultured calli after 6 cycles. This method provides a reference for developing effective transformation protocols for other poplar species.


Asunto(s)
Acetofenonas/metabolismo , Populus/genética , Transformación Genética/genética , Agrobacterium tumefaciens/genética , Vectores Genéticos/genética , Hojas de la Planta/genética , Plantas Modificadas Genéticamente/genética , Reproducibilidad de los Resultados
3.
Biotechnol Bioeng ; 118(1): 153-163, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32897579

RESUMEN

Escherichia coli BL21 (DE3) is an excellent and widely used host for recombinant protein production. Many variant hosts were developed from BL21 (DE3), but improving the expression of specific proteins remains a major challenge in biotechnology. In this study, we found that when BL21 (DE3) overexpressed glucose dehydrogenase (GDH), a significant industrial enzyme, severe cell autolysis was induced. Subsequently, we observed this phenomenon in the expression of 10 other recombinant proteins. This precludes a further increase of the produced enzyme activity by extending the fermentation time, which is not conducive to the reduction of industrial enzyme production costs. Analysis of membrane structure and messenger RNA expression analysis showed that cells could underwent a form of programmed cell death (PCD) during the autolysis period. However, blocking three known PCD pathways in BL21 (DE3) did not completely alleviate autolysis completely. Consequently, we attempted to develop a strong expression host resistant to autolysis by controlling the speed of recombinant protein expression. To find a more suitable protein expression rate, the high- and low-strength promoter lacUV5 and lac were shuffled and recombined to yield the promoter variants lacUV5-1A and lac-1G. The results showed that only one base in lac promoter needs to be changed, and the A at the +1 position was changed to a G, resulting in the improved host BL21 (DE3-lac1G), which resistant to autolysis. As a consequence, the GDH activity at 43 h was greatly increased from 37.5 to 452.0 U/ml. In scale-up fermentation, the new host was able to produce the model enzyme with a high rate of 89.55 U/ml/h at 43 h, compared to only 3 U/ml/h achieved using BL21 (DE3). Importantly, BL21 (DE3-lac1G) also successfully improved the production of 10 other enzymes. The engineered E. coli strain constructed in this study conveniently optimizes recombinant protein overexpression by suppressing cell autolysis, and shows great potential for industrial applications.


Asunto(s)
ARN Polimerasas Dirigidas por ADN/biosíntesis , Regulación hacia Abajo , Escherichia coli , Expresión Génica , Vectores Genéticos , Regiones Promotoras Genéticas , Proteínas Virales/biosíntesis , ARN Polimerasas Dirigidas por ADN/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Virales/genética
4.
Ann Vasc Surg ; 61: 203-211, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31381999

RESUMEN

BACKGROUND: Willis covered stents are used in clinical practice for some complex cerebrovascular diseases. However, the performance of the Willis covered stent requires further investigation. In this study, we investigate the safety and efficacy of Willis covered stents for the treatment of complex vascular diseases of the internal carotid artery (ICA). METHODS: Thirteen patients with complex ICA diseases treated with the Willis covered stent system at our institution from October 2016 to January 2018 were analyzed retrospectively. Follow-up observation and digital subtraction angiography (DSA) examination were conducted at about 6-10 months after the treatment. RESULTS: The complex vascular diseases of the ICA were successfully treated in 12 patients. The technical success rate was 92.3%. Pathologically, 13 lesions included blood blister-like aneurysm (n = 7), traumatic pseudoaneurysm (n = 1), traumatic carotid artery rupture (n = 1), and aneurysm with arteriovenous fistula (n = 4). Thirteen patients with complex vascular diseases of the ICA were treated with 15 Willis covered stents. The release sites of Willis covered stents were the C7 (n = 2), C6 (n = 1), C5 and/or C4 (n = 9), and the C2 (n = 3) segment of the ICA. DSA performed immediately after stent deployment revealed that complete occlusion of the lesion was achieved in 11 patients and endoleak was observed in 2 patients. Of the 11 patients, postoperative DSA examination indicated that the lesions were occluded completely. Among 2 patients, who had a second stent implantation at the break of the ICA, the traumatic ICA rupture was essentially completely obstructed in 1 patient. The endoleak remained in 1 patient with carotid cavernous sinus fistula because the placement of the second stent system was difficult with his ICA tortuosity. No recurrence of aneurysms, hemorrhagia, and other lesions was observed, and the patients' parent arteries were patent without stenosis. No procedure-related complications or deaths occurred during follow-up. CONCLUSIONS: For the treatment of complex vascular diseases in the ICA, Willis covered stent implantation is safe and effective. However, longer follow-up, large-sample controlled studies, and multicenter studies are needed for further confirmation.


Asunto(s)
Enfermedades de las Arterias Carótidas/terapia , Arteria Carótida Interna/fisiopatología , Circulación Cerebrovascular , Círculo Arterial Cerebral/fisiopatología , Procedimientos Endovasculares/instrumentación , Stents , Adolescente , Adulto , Angiografía de Substracción Digital , Enfermedades de las Arterias Carótidas/diagnóstico por imagen , Enfermedades de las Arterias Carótidas/fisiopatología , Arteria Carótida Interna/diagnóstico por imagen , Círculo Arterial Cerebral/diagnóstico por imagen , Procedimientos Endovasculares/efectos adversos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Diseño de Prótesis , Estudios Retrospectivos , Factores de Tiempo , Resultado del Tratamiento , Adulto Joven
5.
J Cell Biochem ; 119(1): 1008-1016, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-28686347

RESUMEN

There was no effective measures can be obtained at present to reverse or prevent airway remodeling. We investigated the therapeutic effect of Erythropoietin (EPO) gene modified mesenchymal stem cells (MSCs) on asthmatic airway remodeling and the possible underlied molecular mechanisms. EPO gene was transfected into MSCs via lentivirus vector. The transfected cells (EPO-MSCs) were identified by flow cytometry and the EPO secreting function was detected by PCR and Western blot. MSCs or EPO-MSCs were administrated to albumin (OVA)-induced chronic asthmatic mouse model via tail veins. The asthmatic phenotype was analyzed. Number of cells in bronchoalveolar lavage fluid (BALF) was counted using a hemocytometer. Histological findings of airways were evaluated by microscopic examination. The concentrations of interleukin 4(IL-4), interleukin 5(IL-5), and interleukin 13(IL-13) in lung homogenate were determined by ELISA. The activation state of transforming growth factor-ß 1 (TGF-ß1), Transforming growth factor beta-activated kinase 1 (TAK1), and p38 Mitogen Activated Protein Kinase (p38MAPK) signaling was detected by Real-Time PCR and Western blotting. EPO-MSCs were successfully constructed. EPO-MSCs showed a more potently suppressive effect on local asthmatic airway inflammation and the level of IL-4, IL-5, and IL-13 in lung tissue than MSCs. Moreover, the numbers of goblet cells, the thicknesses of smooth muscle layer, collagen density, percentage of proliferating cell nuclear antigen positive (PCNA+ ) mesenchymal cells, and von Willebrand factor positive(vWF+ ) vessels were also significantly inhibited by EPO-MSCs. Furthermore, EPO-MSCs could downregulate the expression of TGF-ß1, TAK1, and p38MAPK in lung tissue both in mRNA level and in protein level. EPO gene modified MSCs may more efficiently attenuate asthmatic airway remodeling, which maybe related with the downregulation of TGF-ß1-TAK1-p38MAPK pathway activity.


Asunto(s)
Remodelación de las Vías Aéreas (Respiratorias)/efectos de los fármacos , Asma/terapia , Modelos Animales de Enfermedad , Eritropoyetina/farmacología , Células Madre Mesenquimatosas/citología , Animales , Asma/genética , Asma/metabolismo , Asma/patología , Líquido del Lavado Bronquioalveolar/citología , Eritropoyetina/genética , Regulación de la Expresión Génica , Terapia Genética , Interleucinas/metabolismo , Lentivirus/genética , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C
6.
Planta ; 244(2): 417-27, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27084679

RESUMEN

MAIN CONCLUSION: The PeNAC1 promoter is a non-tissue-specific and stress-inducible promoter containing a GA-responsive element and a MYB recognition sequence that are responsible for induced expression patterns. NAC transcription factors play vital roles in complex signaling networks during plant stress responses. Promoters as crucial molecular switches are involved in the transcriptional regulation of gene activities dynamic network controlling a variety of biological processes, such as developmental processes, responses to hormone and abiotic stress. In this study, a 1217-bp flanking fragment of the stress-responsive NAC gene PeNAC1 was isolated from Populus euphratica. In transgenic Arabidopsis, this promoter fragment was found to have a higher activity than the cauliflower mosaic virus 35S promoter and remained active throughout the plant life cycle, particularly in the spiral vessels and cortical cells of vascular tissues of various organs. We identified a gibberellic acid-responsive element, required for response to gibberellic acid and involved in the salt-stress signaling pathway, and a MYB recognition sequence, which has an important role in promoter response to drought stress, in the PeNAC1 promoter. These results suggest that the PeNAC1 promoter is more effective, non-tissue-specific, and inducible. In addition, the presence of a putative NAC protein-binding motif in the PeNAC1 promoter indicates that PeNAC1 is either regulated by other NAC transcription factors or is self-regulated. Our research will help reveal the regulatory mechanism of the upstream region of the PeNAC1 gene and provide a foundation for the use of the PeNAC1 promoter in molecular breeding.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Populus/genética , Regiones Promotoras Genéticas , Factores de Transcripción/genética , Arabidopsis/genética , Giberelinas/farmacología , Plantas Modificadas Genéticamente/metabolismo , Análisis de Secuencia de ADN , Estrés Fisiológico/genética , beta-Galactosidasa/análisis
7.
Cancer Sci ; 105(6): 660-6, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24673742

RESUMEN

Metastasis is the leading cause of cancer-related death in almost all types of cancers, including colorectal cancer (CRC). Metastasis is a complex, multistep, dynamic biological event, and epithelial-mesenchymal transition (EMT) is a critical process during the cascade. Ajuba family proteins are LIM domain-containing proteins and are reported to be transcription repressors regulating different kinds of physiological processes. However, the expression and pathological roles of Ajuba family proteins in tumors, especial in tumor metastasis, remain poorly studied. Here, we found that JUB, but not the other Ajuba family proteins, was highly upregulated in clinical specimens and CRC cell lines. Ectopic expression of JUB induced EMT and enhanced motility and invasiveness in CRC, and vice versa. Mechanistic study revealed that JUB induces EMT via Snail and JUB is also required for Snail-induced EMT. The expression of JUB shows an inverse correlation with E-cadherin expression in clinical specimens. Taken together, these findings revealed that the LIM protein JUB serves as a tumor-promoting gene in CRC by promoting EMT, a critical process of metastasis. Thus, the LIM protein JUB may provide a novel target for therapy of metastatic CRC.


Asunto(s)
Neoplasias Colorrectales/patología , Transición Epitelial-Mesenquimal , Proteínas con Dominio LIM/metabolismo , Células CACO-2 , Cadherinas/biosíntesis , Movimiento Celular , Neoplasias Colorrectales/genética , Células HCT116 , Humanos , Proteínas con Dominio LIM/genética , Invasividad Neoplásica , Metástasis de la Neoplasia , Interferencia de ARN , ARN Interferente Pequeño , Transducción de Señal , Factores de Transcripción de la Familia Snail , Esferoides Celulares/patología , Factores de Transcripción/metabolismo , Células Tumorales Cultivadas , Regulación hacia Arriba
8.
Respirology ; 19(1): 122-31, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24372635

RESUMEN

BACKGROUND AND OBJECTIVE: Previous studies have demonstrated that our recombinant bacille Calmette-Guerin (rBCG), which expresses Der p2 in house dust mite (Der p2 rBCG) suppresses asthmatic airway inflammation by regulating the phenotype and function of dendritic cells (DC) and reprogramming T helper (Th) 0 cell differentiation into different T cell (Th1/Th2/Treg) subtypes. However, the exact role of Der p2 rBCG in reprogramming Th17 differentiation and the relevant mechanisms are not known. The aim of this study was to examine whether Der p2 rBCG-mediated inhibition of allergic airway inflammation is mediated by regulating Th17 differentiation in a murine asthma model. METHODS: Primary mouse bone marrow-derived dendritic cells (BMDC) were infected with Der p2 rBCG and adoptively transferred to Der p2-intranasally sensitized mice. The role of Der p2 rBCG-BMDC on the regulation of airway inflammation and Th17 cell differentiation was assessed. RESULTS: Adoptive transfer of Der p2 rBCG-BMDC suppressed airway inflammation and mucin secretion. Der p2 rBCG-BMDC inhibited excessive Th17 immune responses but not BCG-BMDC. Furthermore, Der p2 rBCG decreased jagged-2 and increased delta-like-4 expressions on BMDC to a greater extent than BCG. CONCLUSIONS: These findings suggest that DC plays a key role in Der p2 rBCG-induced immunoregulation. Der p2 rBCG also displayed a potent inhibitory effect on Th17 differentiation, and these findings increase our understanding of the cellular basis of Der p2 BCG-mediated inhibition of asthma.


Asunto(s)
Antígenos Dermatofagoides/genética , Proteínas de Artrópodos/genética , Asma/genética , Células de la Médula Ósea/patología , Células Dendríticas/inmunología , Regulación Bacteriana de la Expresión Génica , Mycobacterium bovis/metabolismo , Células Th17/inmunología , Animales , Antígenos Dermatofagoides/biosíntesis , Proteínas de Artrópodos/biosíntesis , Asma/inmunología , Asma/metabolismo , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/microbiología , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Femenino , Ratones , Ratones Endogámicos C57BL , Mycobacterium bovis/inmunología , ARN/genética , Células Th17/metabolismo
9.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(5): 1421-4, 2014 May.
Artículo en Zh | MEDLINE | ID: mdl-25095451

RESUMEN

In M star population, some special objects, which may be of magnetic activity, may be giant stars, or may be of other rare properties, are very important for the follow-up observation and the scientific research on galactic structure and evolution. For local bias of M-type star spectral characteristic lines contained in subspace, a late-type star spectra outlier data mining system is given in the present paper. Firstly, for the sample of M stellar spectral characteristic lines indices, its distribution characteristics in attribute spaces are measured by using the sparse factor and sparsity coefficient, and then this sample is discretized and dimension-reduced to the spectral subspace. Secondly, local outlier subspaces are extracted by PSO (particle swarm optimization) algorithm and identified. Additionally, the effects of sparse coefficient and sparse factor on the number of outliers are discussed by experiments on the sample of SDSS M stellar spectral line index set, and the outliers are compared with spectral type provided by SDSS. In this way, the feasibility and value of this system were validated.

10.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(9): 2424-8, 2014 Sep.
Artículo en Zh | MEDLINE | ID: mdl-25532338

RESUMEN

Pearl is a precious ornament and traditional Chinese medicine, which application history in China is more than 2000 years. It is well known that the chemical ingredients of shell and pearl are very similar, which all of them including calcium carbonate and various amino acids. Generally, shell powders also can be used as medicine; however, its medicinal value is much lower than that of pearl powders. Due to the feature similarity between pearl powders and shell powders, the distinguishment of them by detecting chemical composition and morphology is very difficult. It should be noted that shell powders have been often posing as pearl powders in markets, which seriously infringes the interests of consumers. Identification of pearl powder was investigated by microscopic infrared reflectance spectroscopy, and pearl powder as well as shell powder was calcined at different temperatures for different time before infrared reflectance spectroscopy analysis. The experimental results indicated that when calcined at 400 °C for 30 minutes under atmospheric pressure, aragonite in pearl powder partly transformed into calcite, while aragonite in shell powder completely transformed into calcite. At the same time, the difference in phase transition between the pearl powders 'and shell powders can be easily detected by using the microscopic infrared reflectance spectroscopy. Therefore, based on the difference in their phase transition process, infrared reflectance spectroscopy can be used to identify phase transformation differences between pearl powder and shell powder. It's more meaningfully that the proposed infrared reflectance spec- troscopy method was also investigated for the applicability to other common counterfeits, such as oyster shell powders and abalone shell powders, and the results show that the method can be a simple, efficiently and accurately method for identification of pearl powder.

11.
BMC Cancer ; 13: 412, 2013 Sep 05.
Artículo en Inglés | MEDLINE | ID: mdl-24006921

RESUMEN

BACKGROUND: Colorectal cancer (CRC) is one of the most common cancers worldwide and a leading cause of cancer related death. Although the mortality rate of CRC is decreasing, finding novel targets for its therapy remains urgent. Carboxypeptidase E (CPE), a member of the pro-protein convertases, which are involved in the maturation of protein precursors, has recently been reported as elevated in many types of cancer. However, its role and mechanisms in tumor progression are poorly understood. METHODS: In the present study, we investigated expression of CPE in CRC cell lines and tumor tissues using Western blot and real-time qRT-PCR. Plasmids for overexpression and depletion of CPE were constructed and analyzed by Western blot, MTT and colony formation assays and bromodeoxyuridine incorporation assays. The relative expression of p21, p27, and cyclin D1 were analyzed by Real-time qRT-PCR in the indicated cells. RESULTS: Our study showed that CPE was significantly upregulated in CRC cell lines and tumor tissues. MTT and colony formation assays indicated that overexpression of CPE enhanced cell growth rates. BrdU incorporation and flow-cytometry assays showed that ectopic expression of CPE increased the S-phase fraction cells. Soft agar assay proved enhanced tumorigenicity activity in CPE over-expressing CRC cells. Further studies of the molecular mechanisms of CPE indicated that is promoted cell proliferation and tumorigenicity through downregulation of p21 and p27, and upregulation of cyclin D1. CONCLUSIONS: Taken together, these data suggest that CPE plays an important role in cell cycle regulation and tumorigenicity, and may serve as a potential target for CRC therapeutics.


Asunto(s)
Carboxipeptidasa H/genética , Transformación Celular Neoplásica/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Regulación Neoplásica de la Expresión Génica , Carboxipeptidasa H/metabolismo , Línea Celular Tumoral , Proliferación Celular , Ciclina D1/genética , Ciclina D1/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Fase S/genética , Regulación hacia Arriba
12.
Transfus Apher Sci ; 49(3): 474-81, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23548440

RESUMEN

Transfusion-related acute lung injury (TRALI) is a serious complication associated with blood transfusion and can cause transfusion associated fatalities. Both antibody dependent and non-dependent mechanisms are involved in TRALI, as proposed over the past years. Nonetheless, many details of the immune cells involved in TRALI, particularly the Mac1(+)/Gr1(+) cells from donors, are not fully understood yet. Here we used an in vitro transwell system and a mouse model to study the role of donor leukocytes, present in the donor material, in the occurrence of TRALI reactions. We found that there is a number of immature myeloid cells with Mac1(+)/Gr1(+) phenotype present in the red blood cell (RBC) products, when prepared by regular methods. We found that murine Mac1(+)/Gr1(+) cells from stored RBC products display an elevated MHC I and CD40 expression, as well as an enhanced tumor necrosis factor alpha(TNF-α), interlukin-6(IL-6) and macrophage inflammatory protein 2 (MIP-2) secretion. When tested in a transwell endothelial migration assay, Mac1(+)/Gr1(+) cells showed a significant capability to cross the endothelial barrier. In vivo investigation demonstrated that compared to the purified RBC transfusion, more murine Mac1(+)/Gr1(+) cells from the regular method produced RBC sequestered in the lung, which associated to shorter survival. Taken together, these data suggest that donor derived Mac1(+)/Gr1(+) cells can play a significant role in TRALI reactions, and that reduction of Mac1(+)/Gr1(+) cell number from RBC products is necessary to control the severity of TRALI reactions in clinic.


Asunto(s)
Lesión Pulmonar Aguda/etiología , Células Mieloides/inmunología , Reacción a la Transfusión , Lesión Pulmonar Aguda/inmunología , Adolescente , Adulto , Animales , Anticuerpos/inmunología , Donantes de Sangre , Antígeno CD11b/biosíntesis , Antígeno CD11b/inmunología , Citocinas/inmunología , Femenino , Citometría de Flujo , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Modelos Animales , Receptores de Quimiocina/biosíntesis , Receptores de Quimiocina/inmunología , Adulto Joven
13.
Respiration ; 86(4): 341-6, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23860410

RESUMEN

BACKGROUND: Mucus overproduction is one of the major pathological features of asthma, and MUC5AC is the major mucin component of airway mucus. However, whether Notch signaling is implicated in the regulation of MUC5AC expression in airway secretary cells is still undetermined. OBJECTIVE: The aim of this study is to examine whether Notch signaling can regulate MUC5AC expression and explore the molecular mechanisms. METHODS: Mouse mtCC1-2 cells and human NCI-H292 cells were transfected with NIC, and MUC5AC expression was examined. Using gene reporter assays, site-directed mutagenesis, and ChIP assays, the activity of both mouse and human MUC5AC promoter was analyzed. RESULTS: Notch signaling regulated MUC5AC expression both in mouse mtCC1-2 cells and in human NCI-H292 cells. Several Hes-binding site N-boxes were identified in the 5' region of both mouse and human MUC5AC promoters. Overexpression of NIC resulted in activation of the MUC5AC promoter. Site-directed mutagenesis report assays revealed that Hes proteins might repress both mouse and human MUC5AC promoter activity. Furthermore, ChIP assays confirmed that Hes1 binds to the MUC5AC promoter both in mouse mtCC1-2 cells and in human NCI-H292 cells. CONCLUSIONS: Notch signaling can directly downregulate MUC5AC promoter activity through Hes1-dependent mechanisms, which may be identified as possible targets for pharmacotherapy of airway mucus hypersecretion in asthma.


Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Proteínas de Homeodominio/metabolismo , Mucina 5AC/metabolismo , Receptores Notch/metabolismo , Mucosa Respiratoria/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Línea Celular , Regulación hacia Abajo , Humanos , Ratones , Datos de Secuencia Molecular , Mucina 5AC/genética , Regiones Promotoras Genéticas , Alineación de Secuencia , Factor de Transcripción HES-1
14.
Respiration ; 85(1): 49-58, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23018133

RESUMEN

BACKGROUND: Previous studies showed that a recombinant bacille Calmette-Guérin (rBCG) which expressed the Der p 2 of house dust mites (Der p 2 rBCG) could suppress asthmatic airway inflammation. There are two possible mechanisms: (1) Der p 2 rBCG elicits immune deviation from Th2 to Th1, and (2) Der p 2 rBCG induces antigen-specific regulatory T cells. However, the role of dendritic cell (DC) Der p 2 rBCG in this protective effect and in reprogramming T-cell commitment still needs to be studied. OBJECTIVES: The aim of this study was to determine whether DCs play a central role in the Der p 2 rBCG-mediated inhibition of allergic airway inflammation. METHODS: DCs were collected from Der p 2 rBCG-immunized mice (Der p 2 rBCG-DCs) and adoptively transferred to Der p 2-sensitized mice. The effects of DCs on airway inflammation and immune regulation were analyzed. RESULTS: Adoptive transfer of DCs from Der p 2 rBCG-immunized mice suppressed asthmatic responses, including airway inflammation, mucin secretion and airway responsiveness. Der p 2 rBCG-DCs could effectively inhibit excessive Th2 immune responses and induced a subtype of CD4+CD25+Foxp3+ anti-specific regulatory T cells in this asthma model. Furthermore, Der p 2 rBCG immunization recruited more plasmacytoid DCs in abdominal draining lymph nodes. CONCLUSIONS: These findings suggest that DCs played a key role in Der p 2 rBCG-induced immunoregulation. Compared with BCG, Der p 2 rBCG displayed a more potent inhibitory effect on asthma responses, which may be related to the increase in plasmacytoid DC recruitment. These results improve our understanding of the cellular basis of Der p 2 BCG-mediated inhibition of asthma.


Asunto(s)
Antígenos Dermatofagoides/uso terapéutico , Proteínas de Artrópodos/uso terapéutico , Asma/terapia , Vacuna BCG/uso terapéutico , Células Dendríticas/inmunología , Inmunoterapia Adoptiva/métodos , Inflamación/tratamiento farmacológico , Animales , Antígenos Dermatofagoides/inmunología , Proteínas de Artrópodos/inmunología , Asma/inmunología , Asma/patología , Vacuna BCG/inmunología , Modelos Animales de Enfermedad , Femenino , Inflamación/inmunología , Inflamación/patología , Ratones , Ratones Endogámicos C57BL , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/uso terapéutico , Células TH1/inmunología , Células Th2/inmunología
15.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(8): 2255-8, 2013 Aug.
Artículo en Zh | MEDLINE | ID: mdl-24159888

RESUMEN

Outlier mining is one of the effective methods to find the abnormal celestial spectrum data, and is also one of effective ways to discover the special and unknown celestial bodies. In the present paper, an abnormal characteristic line mining method of celestial spectrum is presented based on the attribute weight and w(k)-distance by using the idea of information entropy. Based on it, an abnormal characteristic line mining system of celestial spectrum was designed and implemented. Firstly, attribute weight of characteristic line was determined by using the idea of information entropy, so that important degree was effectively reflected for each characteristic line. Secondly, massive characteristic line data set of celestial spectrum was reduced by utilizing pruning technique based on neighborhood radius, so that candidate set of abnormal characteristic line was obtained by deleting data objects in which there may not be abnormal characteristic lines. Thirdly, w(k)-distance sum was computed according to the deviation between the data objects in the candidate set, and the objects whose w(k)-distance sum value ranks the first top n were regarded as abnormal characteristic line data objects. In the end, the experimental and the system's running results validated the effectiveness and feasibility of the method by using the SDSS star spectral data set.

16.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(1): 237-40, 2013 Jan.
Artículo en Zh | MEDLINE | ID: mdl-23586264

RESUMEN

Automatic classification and analysis of observational data is of great significance along with the gradual implementation of LAMOST Survey, which will obtain a large number of spectra data. In classification rules extracted, there is often a great deal of redundancy which will reduce the classification efficiency and quality seriously. In the present paper, a post-processing method of star spectra classification rule based on predicate logic is presented by using predication to describe the classification rules and logical reasoning to eliminate redundant rules. In the end, some experimental results on LAMOST's stellar spectra data show that, with no classification accuracy reduction, the efficiency of auto classification is significantly improved.

17.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(10): 2875-8, 2013 Oct.
Artículo en Zh | MEDLINE | ID: mdl-24409754

RESUMEN

Frequent pattern, frequently appearing in the data set, plays an important role in data mining. For the stellar spectrum classification tasks, a classification rule mining method based on classification pattern tree is presented on the basis of frequent pattern. The procedures can be shown as follows. Firstly, a new tree structure, i. e., classification pattern tree, is introduced based on the different frequencies of stellar spectral attributes in data base and its different importance used for classification. The related concepts and the construction method of classification pattern tree are also described in this paper. Then, the characteristics of the stellar spectrum are mapped to the classification pattern tree. Two modes of top-to-down and bottom-to-up are used to traverse the classification pattern tree and extract the classification rules. Meanwhile, the concept of pattern capability is introduced to adjust the number of classification rules and improve the construction efficiency of the classification pattern tree. Finally, the SDSS (the Sloan Digital Sky Survey) stellar spectral data provided by the National Astronomical Observatory are used to verify the accuracy of the method. The results show that a higher classification accuracy has been got.

18.
Respiration ; 83(1): 74-80, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-21968331

RESUMEN

BACKGROUND: Asthmatic airway remodeling is an abnormal injury/repair process of the small airways caused by chronic inflammation in which the quantities of multiple cells increase dramatically. However, the origin of these proliferative cells is still undetermined. OBJECTIVE: The aim of this study was to examine whether bone marrow (BM)-derived adult stem cells are responsible for the proliferative cells in asthmatic airway remodeling. METHODS: Adult mice were durably engrafted with BM isolated from green fluorescent protein (GFP) transgenic mice. Using GFP BM chimera mice, an ovalbumin (OVA)-induced chronic asthma mouse model was established. The distribution of BM-derived GFP+ cells in the lungs of chronic asthma mice was detected by fluorescence microscopy. The phenotype of BM-derived GFP+ cells in the lung tissues of chronic asthma mice was analyzed by flow cytometry. RESULTS: BM chimera mice were successfully generated, with no detectable radioactive inflammation observed. Using BM chimera mice, we established a mouse model of chronic asthma characterized by a significant increase in the thickness of the airway subepithelial basement membrane and smooth muscle layers. OVA treatment caused many GFP+ cells to appear at sites of small airway inflammation. The extravascular localization of some GFP+ cells and their morphology were not consistent with leukocytes. Flow cytometric analysis of lung cells revealed a significant increase in type I collagen (Col I)+GFP+ cells and α-smooth muscle actin (α-SMA)+GFP+ cells in OVA-treated GFP BM chimera mice. CONCLUSIONS: Considerable numbers of Col I- and α-SMA-producing cells originated from BM in the lung tissues of mice with OVA-induced chronic asthma.


Asunto(s)
Remodelación de las Vías Aéreas (Respiratorias) , Asma/patología , Células Madre Hematopoyéticas/fisiología , Hipersensibilidad/patología , Inmunidad Celular , Animales , Asma/inmunología , Células de la Médula Ósea/citología , Células de la Médula Ósea/fisiología , Líquido del Lavado Bronquioalveolar/citología , Diferenciación Celular , Modelos Animales de Enfermedad , Femenino , Citometría de Flujo , Células Madre Hematopoyéticas/citología , Hipersensibilidad/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos
19.
Guang Pu Xue Yu Guang Pu Fen Xi ; 32(5): 1435-8, 2012 May.
Artículo en Zh | MEDLINE | ID: mdl-22827108

RESUMEN

Discretization of continuous numerical attribute is one of the important research works in the preprocessing of celestial spectrum data. For characteristic line of celestial spectrum, a soft discretization algorithm is presented by using improved fuzzy C-means clustering. Firstly, candidate fuzzy clustering centers of characteristic line are chosen by using density values of sample data, so that its anti-noise ability is improved. Secondly, parameters in the fuzzy clustering are dynamically adjusted by taking compatibility of decision table as criteria, so that optimal discretization effect of the characteristic line is achieved. In the end, experimental results effectively validate that the algorithm has higher correct recognition rate of the algorithm by using three SDSS celestial spectrum data sets of high-redshift quasars, late-type star and quasars.

20.
Kaohsiung J Med Sci ; 37(2): 121-127, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33022894

RESUMEN

Cullin 4B (CUL4B) was reported to be closely related to the progression of some tumors, but its function in clear cell renal cell carcinoma (ccRCC) has not been reported. Our present study found CUL4B was upregulated in ccRCC, and CUL4B knockdown markedly inhibited ccRCC cell growth and induced apoptosis. In addition, CUL4B knockdown markedly inhibited antiapoptotic proteins' expression in ccRCC cells, including Mcl-1 and Bcl-2, and silenced CUL4B also induced the cleavages of PARP, an important index of apoptosis. We also confirmed microRNA-217 (miR-217) was downregulated in ccRCC tumor tissues, and negatively correlated with CUL4B expression. Further investigations revealed miR-217 targeted CUL4B and markedly inhibited its expression in ccRCC cells. In addition, overexpression of miR-217 by mimics significantly suppressed ccRCC cell growth. In contrast, enforced expression of CUL4B significantly abolished miR-217-induced cell survival inhibition in ccRCC cells. In conclusion, our present results suggested targeting miR-217-CUL4B axis would be a promising strategy for ccRCC treatment.


Asunto(s)
Apoptosis , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/patología , Proteínas Cullin/metabolismo , Neoplasias Renales/genética , Neoplasias Renales/patología , MicroARNs/metabolismo , Apoptosis/genética , Secuencia de Bases , Línea Celular Tumoral , Proliferación Celular/genética , Supervivencia Celular/genética , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , MicroARNs/genética
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