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1.
Amino Acids ; 2020 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-33245424

RESUMO

As a promising cell therapy, neural crest-derived ectoderm mesenchymal stem cells (EMSCs) secrete high amounts of extracellular matrix (ECM) and neurotrophic factors, promoting neural stem cell (NSC) differentiation into neuronal lineages and aiding tissue regeneration. Additionally, the forced overexpression of secreted proteins can increase the therapeutic efficacy of the secretome. Tissue transglutaminase (TG2) is a ubiquitously expressed member of the transglutaminase family of calcium-dependent crosslinking enzymes, which can stabilize the ECM, inducing smart or living biomaterial to stimulate differentiation and enhance the neurogenesis of NSCs. In this study, we examined the neuronal differentiation of NSCs induced by TG2 gene-modified EMSCs (TG2-EMSCs) in a co-culture model directly. Two weeks after initiating differentiation, levels of the neuronal markers, tubulin beta 3 class III and growth-associated protein 43, were higher in NSCs in the TG2-EMSC co-culture group and those of the astrocytic marker glial fibrillary acidic protein were lower, compared with the control group. These results were confirmed by immunofluorescence, and laminin, fibronectin and sonic hedgehog (Shh) contributed to this effect. The results of western blot analysis and the enzyme-linked immunoassay showed that after TG2-EMSCs were co-cultured for 2 weeks, they expressed much higher levels of Shh than the control group. Moreover, the sustained release of Shh was observed in the TG2-EMSC co-culture group. Overall, our findings indicate that EMSCs can induce the differentiation of NSCs, of which TG2-EMSCs can promote the differentiation of NSCs compared with EMSCs.

2.
Int J Med Mushrooms ; 22(7): 705-717, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32865927

RESUMO

Ganoderma tsugae strain MCCCMAS0053 cultivation on short logs results in varied yield and quality under different growth conditions. Thus, growth conditions need optimization to increase yield and quality. An indoor experiment with three shade treatments (A1, two layers of black sun-shade net; A2, one layer of black sun-shade net plus plastic mulch; A3, one layer of black sun-shade net) and a field experiment in two forest types (pine or mixed pine-oak) were conducted. The results showed that shading and forest type significantly affected light intensities and the growth, size, biomass, and bioactive components of G. tsugae fruiting bodies. In the indoor experiment, the mean dry weight of the fruiting body and the diameter of the pileus in A2 increased by 21.51-44.98% and 13.42-22.26%, respectively, compared with those of A1 and A3. Similarly, the accumulation of the bioactive compounds (polysaccharides, total amino acids, and total essential amino acids) in the pileus and stipe were greater in A2 than in A1 and A3. Furthermore, compared with pure pine forest cultivation, fruiting bodies cultivated in mixed pine-oak forest had greater dry weight, pileus diameter, and more bioactive compounds. In addition, no significant difference was found between the A2 and mixed pine-oak treatments, which had a similar light intensity (from 1116 to 2367 lx). Hence, this suggests that the A2 shade treatment or cultivation in mixed pine-oak forest is beneficial for production of G. tsugae fruiting bodies, and light intensity may play a critical role in this process.

3.
Ann Vasc Surg ; 65: 66-71, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31751599

RESUMO

BACKGROUND: Current methods of treating lower extremity deep venous thrombosis (LEDVT), such as catheter-directed thrombolysis (CDT) alone, or percutaneous mechanical thrombectomy (PMT) alone, are accompanied by unacceptably high risks of complications. This preliminary retrospective study evaluated the efficacy of CDT combined with PMT (via the AngioJet system), relative to CDT alone, in treating LEDVT. METHODS: Forty-two patients (43 limbs) with symptomatic deep venous thrombosis received either CDT alone (n = 12) or PMT combined with CDT (PMT + CDT) from May 2012 to December 2016. The groups were compared for clinical outcomes and demographics, LEDVT risk factors, and dosages of urokinase. Thrombus removal, by venographic evidence, was classified as grades I (<50%), II (50 to 99%), or III (>99%). RESULTS: In the CDT (PMT + CDT) cohorts, grades I, II, and III thrombus removal was achieved by 8% (3%), 17% (10%), and 75% (87%) of patients, respectively. The urokinase dosage and hospitalization required by the CDT group (5.29 ± 0.45 million IU, 20.4 ± 4.6 days) were significantly greater than those required by the PMT + CDT group (4.08 ± 1.15 million IU, 16.0 ± 6.0 days; P = 0.001, 0.039). The clinical outcomes of the 2 groups were similar. CONCLUSION: Combined PMT and CDT was effective and safe for LEDVT clinical therapy, and hospital stay, urokinase dosage, and complications were less compared with patients who received CDT only.


Assuntos
Fibrinolíticos/administração & dosagem , Extremidade Inferior/irrigação sanguínea , Trombectomia/instrumentação , Terapia Trombolítica , Ativador de Plasminogênio Tipo Uroquinase/administração & dosagem , Trombose Venosa/terapia , Adulto , Idoso , Terapia Combinada , Feminino , Fibrinolíticos/efeitos adversos , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Trombectomia/efeitos adversos , Terapia Trombolítica/efeitos adversos , Fatores de Tempo , Resultado do Tratamento , Ativador de Plasminogênio Tipo Uroquinase/efeitos adversos , Grau de Desobstrução Vascular , Trombose Venosa/diagnóstico por imagem , Trombose Venosa/fisiopatologia
4.
BMC Genomics ; 20(1): 880, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31747891

RESUMO

BACKGROUND: Melatonin is a ubiquitous molecule and exists across kingdoms. Studies on melatonin in plants have mainly focused on its physiological influence on growth and development, and on its biosynthesis. A number of studies have been conducted on the melatonin content and exogenous melatonin treatment of grapevine (Vitis vinifera L.). However, key genes or enzymes of the melatonin biosynthetic pathway remain unclear. RESULTS: In this study, we cloned and identified the gene encoding serotonin N-acetyltransferase (SNAT) in grapevine (VvSNAT2). The VvSNAT2 protein was identified from a collection of 30 members of the grapevine GCN5-related N-acetyltransferase (GNAT) superfamily. Phylogenetic and protein sublocalization analyses showed that the candidate gene VvGNAT16 is VvSNAT2. Characterization of VvSNAT2 showed that its enzymatic activity is highest at a pH of 8.8 and a temperature of 45 °C. Analysis of enzyme kinetics showed the values of Km and Vmax of VvSNAT2 using serotonin were 392.5 µM and 836 pmol/min/mg protein, respectively. The expression of VvSNAT2 was induced by melatonin treatment and pathogen inoculation. Overexpression of VvSNAT2 in Arabidopsis resulted in greater accumulation of melatonin and chlorophyll and enhanced resistance to powdery mildew in the transgenic plants compared with the wild type (WT). Additionally, our data showed that the marker genes in the salicylic acid (SA) signaling pathway were expressed to higher levels in the transgenic plants compared with the WT. CONCLUSIONS: The VvSNAT2 gene was cloned and identified in grapevine for the first time. Our results indicate that VvSNAT2 overexpression activates the SA and JA signaling pathways; however, the SA pathway plays a central role in VvSNAT2-mediated plant defense.


Assuntos
Arilalquilamina N-Acetiltransferase/genética , Regulação da Expressão Gênica de Plantas , Melatonina/biossíntese , Doenças das Plantas/genética , Proteínas de Plantas/genética , Vitis/genética , Arabidopsis/enzimologia , Arabidopsis/genética , Arabidopsis/imunologia , Arilalquilamina N-Acetiltransferase/imunologia , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/patogenicidade , Clorofila/biossíntese , Clonagem Molecular , Ciclopentanos/metabolismo , Expressão Gênica , Oxilipinas/metabolismo , Filogenia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/imunologia , Plantas Geneticamente Modificadas , Plasmídeos/química , Plasmídeos/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Ácido Salicílico/metabolismo , Serotonina/metabolismo , Transdução de Sinais , Vitis/classificação , Vitis/enzimologia , Vitis/imunologia
5.
Plant Physiol Biochem ; 143: 183-189, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31513952

RESUMO

DOF proteins are plant-specific transcription factors that play vital roles in plant development and defense responses. However, DOFs have primarily been investigated in model plants, and fairly limited research has been performed on grape (Vitis vinifera). In this study, we isolated and characterized a C2-C2 zinc finger structural DOF gene, VvDOF3, from the grape cultivar Jingxiu. The VvDOF3 protein showed nuclear localization and transcriptional activation ability, indicating that it functions as a transcription factor. The VvDOF3 gene was rapidly induced by exogenous salicylic acid (SA), jasmonic acid (JA), and powdery mildew infection. Overexpression of VvDOF3 in Arabidopsis thaliana enhanced resistance to Golovinomyces cichoracearum. Expression of the SA-responsive defense-related gene PR1 and the concentration of SA were up-regulated in transgenic Arabidopsis plants overexpressing VvDOF3. Together, these data suggest that VvDOF3 functions as a transcription factor in grape and enhances powdery mildew resistance through the SA signaling pathway.


Assuntos
Proteínas de Plantas/metabolismo , Vitis/microbiologia , Ciclopentanos/farmacologia , Resistência à Doença/genética , Resistência à Doença/fisiologia , Oxilipinas/farmacologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Ácido Salicílico/farmacologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Vitis/metabolismo
6.
Int J Mol Sci ; 20(17)2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31480584

RESUMO

Powdery mildew is a disease caused by fungal pathogens that harms grape leaves and fruits. The TIFY gene family is a plant-specific super-family involved in the process of plants' development and their biotic and abiotic stress responses. This study aimed to learn the function of the VvTIFY9 gene to investigate molecular mechanisms of grape resistance to powdery mildew. A VvTIFY9 protein encoding a conserved motif (TIF[F/Y]XG) was characterized in grape (Vitis vinifera). Sequence analysis confirmed that VvTIFY9 contained this conserved motif (TIF[F/Y]XG). Quantitative PCR analysis of VvTIFY9 in various grape tissues demonstrated that the expression of VvTIFY9 was higher in grape leaves. VvTIFY9 was induced by salicylic acid (SA) and methyl jasmonate (MeJA) and it also quickly responded to infection with Erysiphe necator in grape. Analysis of the subcellular localization and transcriptional activation activity of VvTIFY9 showed that VvTIFY9 located to the nucleus and had transcriptional activity. Arabidopsis that overexpressed VvTIFY9 were more resistant to Golovinomyces cichoracearum, and quantitative PCR revealed that two defense-related genes, AtPR1 and AtPDF1.2, were up-regulated in the overexpressing lines. These results indicate that VvTIFY9 is intimately involved in SA-mediated resistance to grape powdery mildew. This study provides the basis for exploring the molecular mechanism of grape resistance to disease resistance and candidate genes for transgenic disease resistance breeding of grape plants.


Assuntos
Resistência à Doença , Micoses/metabolismo , Doenças das Plantas , Fatores de Transcrição/metabolismo , Vitis/metabolismo , Acetatos/metabolismo , Sequência de Aminoácidos , Ascomicetos , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Ácido Salicílico/metabolismo , Análise de Sequência de Proteína , Estresse Fisiológico , Fatores de Transcrição/química , Fatores de Transcrição/fisiologia , Vitis/fisiologia
7.
Mol Biotechnol ; 61(11): 816-825, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31486973

RESUMO

Methyl parathion hydrolase (MPH) plays an important role in degrading a range of organophosphorus compounds. In order to display MPH on the cell surface of Escherichia coli strain RosettaBlue™, the Flavin-based fluorescent protein EcFbFP was severed as an auto-anchoring matrix. With net negative charges of EcFbFP supplying the driving forces, fusion protein MPH-EcFbFP through a two-step auto-surface display process was finally verified by (a) inner membrane translocation and (b) anchoring at outer membrane. Cells with surface-displayed MPH obtained activity of 0.12 U/OD600 against substrate methyl parathion. MPH when fused with engineered EcFbFP containing 20 net negative charges exhibited fivefold higher anchoring efficiency and tenfold higher enzymatic catalytic activity of 1.10 U/OD600. The above result showed that MPH was successfully displayed on cell surface and can be used for biodegradation of methyl parathion.


Assuntos
Escherichia coli/genética , Hidrolases/genética , Proteínas Luminescentes/genética , Metil Paration/metabolismo , Biocatálise , Biodegradação Ambiental , Membrana Celular/metabolismo , Técnicas de Visualização da Superfície Celular , Escherichia coli/enzimologia , Escherichia coli/metabolismo , Mononucleotídeo de Flavina , Hidrolases/metabolismo , Proteínas Luminescentes/química , Compostos Organofosforados/metabolismo , Proteínas Recombinantes de Fusão
8.
Pathol Res Pract ; 215(10): 152555, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31445715

RESUMO

Recent evidence highlights that microRNAs serve as crucial regulators of tumorigenesis, including non-small cell lung cancer (NSCLC). The present study was designed to investigate the expression profile, clinical significance and biological role of miR-421 in NSCLC. The results showed that miR-421 expression was markedly increased in NSCLC tissues and cell lines. Further experimental data indicated that knockdown of miR-421 significantly inhibited NSCLC cell proliferation and induced cell cycle arrest in vitro. The migratory and invasive abilities of NSCLC cells were also attenuated following miR-421 knockdown. Furthermore, PDCD4 was identified as a direct target of miR-421, and its expression was inversely correlated with miR-421 expression in NSCLC tissues. PDCD4 also abrogated the oncogenic role of miR-421 in NSCLC cells. Collectively, our study revealed that miR-421 is significantly upregulated in NSCLC and might represent a potential therapeutic target for NSCLC patients.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Proliferação de Células/fisiologia , Neoplasias Pulmonares/metabolismo , Pulmão/metabolismo , MicroRNAs/metabolismo , Invasividade Neoplásica/patologia , Proteínas de Ligação a RNA/metabolismo , Idoso , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Pulmão/patologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica/genética
9.
Exp Cell Res ; 361(1): 56-62, 2017 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-28986067

RESUMO

Senior individuals older than 65 years of age are at a disproportionally higher risk of developing pneumonia. Impaired capacity to defend against airway infections may be one of the reasons. It is generally believed that weaker regulatory T cell responses may be beneficial to host defense against pathogens. In senior patients with community-acquired bacterial pneumonia, we investigated the frequencies and functions of regulatory T cells. Interestingly, we found that compared to age- and sex-matched healthy controls, senior pneumonia patients presented lower frequencies of Foxp3-expressing and Helios-expressing CD4+ T cells. The quantity of Foxp3 and Helios being expressed, measured by their mRNA transcription levels, was also lower in CD4+ T cells from pneumonia patients. Furthermore, following TCR and TGF-ß stimulation, pneumonia patients presented impaired capacity to upregulate Foxp3 and Helios. Functional analyses revealed that CD4+ T cells from pneumonia patients secreted lower amounts of IL-10 and TGF-ß, two cytokines critical to regulatory T cell-mediated suppression. Also, the expression of granzyme B and perforin, which were cytolytic molecules potentially utilized by regulatory T cells to mediate the elimination of antigen-presenting cells and effector T cells, were reduced in CD4+CD25+ T cells from senior pneumonia patients. In addition, the CD4+CD25+ T cells from senior pneumonia patients presented reduced capacity to suppress effector CD4+ and CD8+ T cell proliferation. Moreover, the value of pneumonia severity index was inversely correlated with several parameters of regulatory T cell function. Together, our results demonstrated that senior pneumonia patients presented a counterintuitive impairment in regulatory T cell responses that was associated with worse prognosis.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Infecções Comunitárias Adquiridas/imunologia , Pneumonia Bacteriana/imunologia , Índice de Gravidade de Doença , Linfócitos T Reguladores/imunologia , Idoso , Estudos de Casos e Controles , Infecções Comunitárias Adquiridas/patologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Pneumonia Bacteriana/patologia
10.
APMIS ; 125(12): 1108-1116, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28913840

RESUMO

Pneumoconiosis is caused by the accumulation of airborne dust in the lung, which stimulates a progressive inflammatory response that ultimately results in lung fibrosis and respiratory failure. It is possible that regulatory cells in the immune system could function to suppress inflammation and possibly slow or reverse disease progression. However, results in this study suggest that in pneumoconiosis patients, the regulatory T cells (Tregs) and B cells are functionally impaired. First, we found that pneumoconiosis patients presented an upregulation of CD4+ CD25+ T cells compared to controls, whereas the CD4+ CD25+ and CD4+ CD25hi T cells were enriched with Th1- and Th17-like cells but not Foxp3-expressing Treg cells and evidenced by significantly higher T-bet, interferon (IFN)-γ, and interleukin (IL)-17 expression but lower Foxp3 and transforming growth factor (TGF)-ß expression. Regarding the CD4+ CD25hi T-cell subset, the frequency of this cell type in pneumoconiosis patients was significantly reduced compared to controls, together with a reduction in Foxp3 and TGF-ß and an enrichment in T-bet, RORγt, IFN-γ, and IL-17. This skewing toward Th1 and Th17 types of inflammation could be driven by monocytes and B cells, since after depleting CD14+ monocytes and CD19+ B cells, the levels of IFN-γ and IL-17 were significantly decreased. Whole peripheral blood mononuclear cells and isolated monocytes and B cells in pneumoconiosis patients also presented reduced capacity of TGF-ß secretion. Furthermore, monocytes and B cells from pneumoconiosis patients presented reduced capacity in inducing Foxp3 upregulation, a function that could be rescued by exogenous TGF-ß. Together, these data indicated a potential pathway for the progression of pneumoconiosis through a loss of Foxp3+ Treg cells associated with impaired TGF-ß secretion.


Assuntos
Pneumoconiose/imunologia , Linfócitos T Reguladores/imunologia , Fator de Crescimento Transformador beta/biossíntese , Idoso , Linfócitos B/imunologia , Estudos de Casos e Controles , Citocinas/genética , Progressão da Doença , Fatores de Transcrição Forkhead/metabolismo , Humanos , Tolerância Imunológica , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Pneumoconiose/etiologia , Pneumoconiose/genética , Proteínas Proto-Oncogênicas c-bcl-6/metabolismo , Proteínas com Domínio T/metabolismo , Linfócitos T Reguladores/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo
11.
Mycologia ; 109(1): 64-74, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28402798

RESUMO

Three new species of Coltricia from southern China are described on the basis of morphological characters and molecular analyses. Phylogenetic analyses based on nuc rDNA ITS1-5.8S-ITS2 and partial 28S rDNA, and partial sequences of genes for RNA polymerase II subunits 1 and 2 confirmed the generic placement of the three new species. Coltricia lateralis is characterized by small, laterally stipitate basidiocarps, obtuse pileus margin, cinnamon-buff to honey yellow pore surface, and broadly ellipsoid basidiospores that measure 7-8 × 5-6 µm. Coltricia rigida is characterized by large, woody-hard fan-shaped basidiocarps, small pores (7-8 per mm), presence of septate sclerified hyphae, and subglobose to globose basidiospores that measure 6-7 × 5-6.5 µm. Coltricia wenshanensis is characterized by centrally stipitate basidiocarps, a distinctly concentrically zonate and sulcate pileal surface, large pores (0.5-2 per mm), and broadly ellipsoid to subglobose basidiospores that measure 7.5-8 × 6-7 µm.


Assuntos
Basidiomycota/classificação , Basidiomycota/isolamento & purificação , Basidiomycota/citologia , Basidiomycota/genética , China , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microscopia , Filogenia , RNA Polimerase II/genética , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
12.
PLoS One ; 11(8): e0160367, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27479442

RESUMO

Ice nucleation protein (INP) is frequently used as a surface anchor for protein display in gram-negative bacteria. Here, MalE and TorA signal peptides, and three charged polypeptides, 6×Lys, 6×Glu and 6×Asp, were anchored to the N-terminus of truncated INP (InaK-N) to improve its surface display efficiency for human Arginase1 (ARG1). Our results indicated that the TorA signal peptide increased the surface translocation of non-protein fused InaK-N and human ARG1 fused InaK-N (InaK-N/ARG1) by 80.7% and 122.4%, respectively. Comparably, the MalE signal peptide decreased the display efficiencies of both the non-protein fused InaK-N and InaK-N/ARG1. Our results also suggested that the 6×Lys polypeptide significantly increased the surface display efficiency of K6-InaK-N/ARG1 by almost 2-fold, while also practically abolishing the surface translocation of non-protein fused InaK-N, indicating the interesting roles of charged polypeptides in bacteria surface display systems. Cell surface-immobilized K6-InaK-N/ARG1 presented an arginase activity of 10.7 U/OD600 under the optimized conditions of 40°C, pH 10.0 and 1 mM Mn2+, which could convert more than 95% of L-Arginine (L-Arg) to L-Ornithine (L-Orn) in 16 hours. The engineered InaK-Ns expanded the INP surface display system, which aided in the surface immobilization of human ARG1 in E. coli cells.


Assuntos
Arginase/metabolismo , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/metabolismo , Escherichia coli/metabolismo , Arginase/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Citometria de Fluxo , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Humanos , Lisina/genética , Lisina/metabolismo , Microscopia de Fluorescência , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Ornitina/metabolismo , Engenharia de Proteínas , Sinais Direcionadores de Proteínas/genética , Translocação Genética
13.
Appl Phys Lett ; 108(21): 213901, 2016 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-27375297

RESUMO

This work introduces a type of CdS/CuxS quantum dots (QDs) as sensitizers in quantum dot sensitized solar cells by in-situ cationic exchange reaction method where CdS photoanode is directly immersed in CuCl2 methanol solution to replace Cd2+ by Cu2+. The p-type CuxS layer on the surface of the CdS QDs can be considered as hole transport material, which not only enhances the light harvesting of photoanode but also boosts the charge separation after photo-excitation. Therefore, both the electron collection efficiency and power conversion efficiency of the solar cell are improved from 80% to 92% and from 1.21% to 2.78%, respectively.

14.
Int J Occup Med Environ Health ; 28(1): 62-70, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26159948

RESUMO

INTRODUCTION: Information on the changes of pulmonary function and the right cardiac structure in patients with coal worker's pneumoconiosis in China is very scarce. This study was performed to clarify the changes of pulmonary function and right cardiac structure in patients with coal worker's pneumoconiosis in China. MATERIAL AND METHODS: Pulmonary function, pulmonary artery systolic pressure, and the right cardiac structure were evaluated by spirometry and color Doppler echocardiography. RESULTS: The pulmonary artery systolic pressure of patients with coal worker's pneumoconiosis was increased with disease severity. Patients with coal worker's pneumoconiosis also exhibited an impaired pulmonary function and altered right cardiac structure compared with control subjects. A significant linear correlation of the variables of pulmonary ventilation and diffusion function with the indicators of the right cardiac structure was found in patients with coal worker's pneumoconiosis in China. CONCLUSIONS: This study elucidated a deterioration of pulmonary function and right cardiac structure in patients with coal worker's pneumoconiosis in China.


Assuntos
Antracose/fisiopatologia , Átrios do Coração/diagnóstico por imagem , Ventrículos do Coração/diagnóstico por imagem , Idoso , Pressão Arterial , China , Ecocardiografia Doppler , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Artéria Pulmonar , Índice de Gravidade de Doença , Espirometria , Sístole
15.
PLoS One ; 10(7): e0131757, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26134129

RESUMO

The present study assessed high-level expression of the KOD DNA polymerase in Pichia pastoris. Thermococcus kodakaraensis KOD1 is a DNA polymerase that is widely used in PCR. The DNA coding sequence of KOD was optimized based on the codon usage bias of P. pastoris and synthesized by overlapping PCR, and the nonspecific DNA-binding protein Sso7d from the crenarchaeon Sulfolobus solfataricus was fused to the C-terminus of KOD. The resulting novel gene was cloned into a pHBM905A vector and introduced into P. pastoris GS115 for secretory expression. The yield of the target protein reached approximately 250 mg/l after a 6-d induction with 1% (v/v) methanol in shake flasks. This yield is much higher than those of other DNA polymerases expressed heterologously in Escherichia coli. The recombinant enzyme was purified, and its enzymatic features were studied. Its specific activity was 19,384 U/mg. The recombinant KOD expressed in P. pastoris exhibited excellent thermostability, extension rate and fidelity. Thus, this report provides a simple, efficient and economic approach to realize the production of a high-performance thermostable DNA polymerase on a large scale. This is the first report of the expression in yeast of a DNA polymerase for use in PCR.


Assuntos
Proteínas Arqueais/genética , Proteínas de Ligação a DNA/genética , DNA Polimerase Dirigida por DNA/biossíntese , DNA Polimerase Dirigida por DNA/genética , Proteínas Fúngicas/biossíntese , Proteínas Fúngicas/genética , Pichia/enzimologia , Proteínas Arqueais/biossíntese , Códon , DNA/química , Proteínas de Ligação a DNA/biossíntese , DNA Polimerase Dirigida por DNA/química , Escherichia coli/enzimologia , Glicoproteínas/química , Fases de Leitura Aberta , Reação em Cadeia da Polimerase , Estrutura Terciária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Análise de Sequência de DNA , Temperatura , Thermococcus/enzimologia
16.
Artigo em Inglês | MEDLINE | ID: mdl-25703950

RESUMO

A sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine kurarinone in rat plasma using chlorzoxazone as the internal standard (IS). Sample preparation was accomplished through a liquid-liquid extraction procedure with ethyl acetate to 0.2 mL plasma sample. The analyte and IS were separated on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 µm) with the mobile phase of acetonitrile and 1% formic acid in water with gradient elution at a flow rate of 0.40 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) by multiple reactions monitoring (MRM) of the transitions at m/z 437.0→301.2 for kurarinone and m/z 168.1→132.1 for IS. The linearity of this method was found to be within the concentration range of 20-2000 ng/mL with a lower limit of quantification of 20 ng/mL. Only 3.0 min was needed for an analytical run. The matrix effect was 94.7-107.2% for kurarinone. The intra- and inter-day precision (RSD%) were less than 8.2% and accuracy (RE%) was within ±9.0%. The recovery ranged from 77.3% to 85.6%. Kurarinone was sufficiently stable under all relevant analytical conditions. The method was also successfully applied to the pharmacokinetic study of kurarinone in rats.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/sangue , Espectrometria de Massas em Tandem/métodos , Administração Intravenosa , Animais , Estabilidade de Medicamentos , Flavonoides/administração & dosagem , Flavonoides/química , Flavonoides/farmacocinética , Modelos Lineares , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
17.
J Insect Sci ; 14: 253, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25434038

RESUMO

Musca domestica L. (Diptera: Muscidae), the housefly, exhibits unique immune defenses and can produce antibacterial substances upon stimulation with bacteria. On the basis of the cDNA library constructed using the suppression subtractive hybridization method, a 1188-bp antibacterial substance gene, which we named AS566, was amplified by rapid amplification of cDNA ends from M. domestica larva stimulated with Salmonella pullorum (Enterobacteriaceae: Salmonella). In this study, the full-length AS566 gene was cloned and inserted into a His-tagged Escherichia coli (Enterobacteriaceae: Escherichia) prokaryotic expression system to enable production of the recombinant protein. The recombinant AS566 protein was purified in denatured form from inclusion bodies and renatured to obtain functionally active AS566 protein. The bacteriostatic activity of the recombinant purified AS566 protein was assessed using the Oxford plate assay system and the results indicated that AS566 had antibacterial activity against six bacteria, including an E. coli clinical isolate, S. pullorum, Streptococcus bovis (Streptococcaceae: Streptococcus), Streptococcus suis, and Staphylococcus aureus (Staphylococcaceae: Staphylococcus) in vitro. The antibacterial activity of AS566 toward Gram- bacteria was two times greater than that against Gram+ bacteria. The sequencing results and BLAST analysis showed that the antibacterial substance gene AS566 was not homologous to any other antibacterial substance genes in GenBank. The antibacterial mechanisms of the newly discovered AS566 protein warrant further study.


Assuntos
Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Moscas Domésticas/genética , Proteínas de Insetos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Biblioteca Gênica , Moscas Domésticas/crescimento & desenvolvimento , Moscas Domésticas/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
18.
Am J Ind Med ; 57(8): 866-71, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24899082

RESUMO

BACKGROUND: Coal workers' pneumoconiosis (CWP) is characterized by progressive pulmonary fibrosis. Inflammation is crucial in the host response to silica and it contributes to pulmonary fibrosis. Cyclooxygenase-2 is involved in this process. The association between single-nucleotide polymorphisms in cyclooxygenase-2 gene with pneumoconiosis risk was investigated. METHODS: An association study was conducted by analyzing two single-nucleotide polymorphisms of cyclooxygenase-2 (rs689466 and rs20417) in a case-control study involving 90 patients and 90 healthy individuals (controls). Genotyping was performed by the TaqMan method. RESULTS: The rs689466 AG and GG and rs689466 GC polymorphisms were significantly less frequent in patients than in controls. Cyclooxygenase-2 rs689466 and rs20417 variant genotypes exhibited 21% and 12% decreased CWP risk, respectively. CONCLUSION: Cyclooxygenase-2 rs689466 and rs20417 polymorphisms were associated with CWP risk.


Assuntos
Antracose/genética , Ciclo-Oxigenase 2/genética , Polimorfismo de Nucleotídeo Único , Alelos , Estudos de Casos e Controles , Genótipo , Humanos , Pessoa de Meia-Idade , Fatores de Risco
19.
Huan Jing Ke Xue ; 34(10): 3840-6, 2013 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-24364301

RESUMO

Runoff samples of the Shanghai elevated inner highway were collected and analyzed during the rainy season of 2012 to investigate the pollution characteristics of 16 USEPA priority pollutant polycyclic aromatic hydrocarbons (PAHs). Total PAHs concentrations ranged from 1.585 to 7.523 microg.L-1. The sources of PAHs in the runoff were determined by using 2 source apportionment models, positive matrix factorization (PMF) and principal component analysis with multivariate linear regression (PCA/ MLR). Source profiles and contributions obtained from the two models were comparable. For PMF, PAHs mainly came from vehicle emissions (37. 7% ), followed by petroleum products (21. 9% ) , combustive gas (26. 4%), and other sources (14.0% ). As for PCA/MLR, it was 44. 3% , 28.9% , 18. 3% , 8.5% , respectively. The relation between the modeled total PAH concentrations and the measured concentrations was strong, with the correlation coefficient of 0. 961 for PMF and 0. 997 for PCA/MLR.


Assuntos
Monitoramento Ambiental , Poluição Ambiental/análise , Modelos Teóricos , Hidrocarbonetos Policíclicos Aromáticos/análise , China , Petróleo , Análise de Componente Principal , Chuva , Estações do Ano , Emissões de Veículos
20.
Artigo em Chinês | MEDLINE | ID: mdl-23803542

RESUMO

OBJECTIVE: To investigate the effects of whole lung lavage (WLL) on the pulmonary function and exercise capacity in patients with pneumoconiosis. METHODS: Forty-one patients with pneumoconiosis who quit dust-exposed work not more than 6 months before underwent WLL. Clinical symptom assessment, pulmonary function test, and cardiopulmonary exercise test were performed before and one week after WLL, and the results were compared. RESULTS: The patients with pneumoconiosis showed no significant changes in clinical symptoms after WLL. At one week after WLL, the patients with pneumoconiosis showed nonsignificant increases in forced vital capacity, forced expiratory volume in one second (FEV1.0), and percent predicted FEV1 (P > 0.05); peak oxygen uptake (peak VO2) increased from 2140.6 ± 353.2 ml/min before WLL to 2374.6 ± 362.4 ml/min after WLL, percent predicted peak VO2 increased from 82.2 ± 13.7% before WLL to 91.0 ± 14.0% after WLL, peak VO2/kg increased from 30.6 ± 3.5 ml/min×kg before WLL to 34.2 ± 3.7 ml/min×kg after WLL, and ventilatory equivalent for carbon dioxide decreased from 30.6 ± 3.1 before WLL to 26.1 ± 2.7 after WLL (P < 0.05). CONCLUSION: WLL can remarkably improve the oxygen uptake and ventilatory efficiency in patients with pneumoconiosis during exercise, so it can improve the exercise capacity of these patients.


Assuntos
Lavagem Broncoalveolar , Tolerância ao Exercício , Pulmão/fisiopatologia , Pneumoconiose/terapia , Adulto , Humanos , Pessoa de Meia-Idade , Pneumoconiose/fisiopatologia
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