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1.
J Cell Sci ; 136(15)2023 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-37461827

RESUMEN

Protein palmitoylation is a post-translational lipid modification of proteins. Accumulating evidence reveals that palmitoylation functions as a sorting signal to direct proteins to destinations; however, the sorting mechanism remains largely unknown. Here, we show that ARF6 plays a general role in targeting palmitoylated proteins from the Golgi to the plasma membrane (PM). Through shRNA screening, we identified ARF6 as the key small GTPase in targeting CD36, a palmitoylated protein, from the Golgi to the PM. We found that the N-terminal myristoylation of ARF6 is required for its binding with palmitoylated CD36, and the GTP-bound form of ARF6 facilitates the delivery of CD36 to the PM. Analysis of stable isotope labeling by amino acids in cell culture revealed that ARF6 might facilitate the sorting of 359 of the 531 palmitoylated PM proteins, indicating a general role of ARF6. Our study has thus identified a sorting mechanism for targeting palmitoylated proteins from the Golgi to the PM.


Asunto(s)
Aparato de Golgi , Proteínas de la Membrana , Membrana Celular/metabolismo , Aparato de Golgi/metabolismo , Proteínas de la Membrana/metabolismo , Transporte de Proteínas
2.
Proc Natl Acad Sci U S A ; 118(21)2021 05 25.
Artículo en Inglés | MEDLINE | ID: mdl-34001600

RESUMEN

G-quadruplexes (G4s) formed by guanine-rich nucleic acids play a role in essential biological processes such as transcription and replication. Besides the >1.5 million putative G-4-forming sequences (PQSs), the human genome features >640 million single-nucleotide variations (SNVs), the most common type of genetic variation among people or populations. An SNV may alter a G4 structure when it falls within a PQS motif. To date, genome-wide PQS-SNV interactions and their impact have not been investigated. Herein, we present a study on the PQS-SNV interactions and the impact they can bring to G4 structures and, subsequently, gene expressions. Based on build 154 of the Single Nucleotide Polymorphism Database (dbSNP), we identified 5 million gains/losses or structural conversions of G4s that can be caused by the SNVs. Of these G4 variations (G4Vs), 3.4 million are within genes, resulting in an average load of >120 G4Vs per gene, preferentially enriched near the transcription start site. Moreover, >80% of the G4Vs overlap with transcription factor-binding sites and >14% with enhancers, giving an average load of 3 and 7.5 for the two regulatory elements, respectively. Our experiments show that such G4Vs can significantly influence the expression of their host genes. These results reveal genome-wide G4Vs and their impact on gene activity, emphasizing an understanding of genetic variation, from a structural perspective, of their physiological function and pathological implications. The G4Vs may also provide a unique category of drug targets for individualized therapeutics, health risk assessment, and drug development.


Asunto(s)
Proteínas de Unión al ADN/ultraestructura , G-Cuádruplex , Genoma Humano/genética , Conformación de Ácido Nucleico , Proteínas de Unión al ADN/genética , Regulación de la Expresión Génica/genética , Humanos , Polimorfismo de Nucleótido Simple/genética , Regiones Promotoras Genéticas/genética , Unión Proteica/genética , Secuencias Reguladoras de Ácidos Nucleicos/genética , Sitio de Iniciación de la Transcripción , Activación Transcripcional/genética
3.
Zhongguo Zhong Yao Za Zhi ; 48(19): 5365-5376, 2023 Oct.
Artículo en Zh | MEDLINE | ID: mdl-38114126

RESUMEN

The present study aimed to explore the underlying mechanism of Wuling Capsules in the treatment of hepatic fibrosis(HF) through network pharmacology, molecular docking, and animal experiments. Firstly, the chemical components and targets of Wuling Capsules against HF were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), Traditional Chinese Medicines Integrated Database(TCMID), GeneCards, and literature retrieval. The protein-protein interaction(PPI) network analysis was carried out on the common targets by STRING database and Cytoscape 3.9.1 software, and the core targets were screened, followed by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. Enrichment analysis was conducted on the core targets and the "drug-core component-target-pathway-disease" network was further constructed. Subsequently, molecular docking between core components and core targets was conducted using AutoDock Vina software to predict the underlying mechanism of action against HF. Finally, an HF model induced by CCl_4 was constructed in rats, and the general signs and liver tissue morphology were observed. HE and Masson staining were used to analyze the liver tissue sections. The effects of Wuling Capsules on the levels of inflammatory factors, hydroxyproline(HYP) levels, and core targets were analyzed by ELISA, RT-PCR, etc. A total of 445 chemical components of Wuling Capsules were screened, corresponding to 3 882 potential targets, intersecting with 1 240 targets of HF, and 47 core targets such as TNF, IL6, INS, and PIK3CA were screened. GO and KEGG enrichment analysis showed that the core targets mainly affected the process of cell stimulation response and metabolic regulation, involving cancer, PI3K-Akt, MAPK, and other signaling pathways. Molecular docking showed that the core components of Wuling Capsules, such as lucidenic acid K, ganoderic acid B, lucidenic acid N, saikosaponin Q2, and neocryptotanshinone, had high affinities with the core targets, such as TNF, IL6 and PIK3CA. Animal experiments showed that Wuling Capsules could reduce fat vacuole, inflammatory infiltration, and collagen deposition in rat liver, decrease the levels of inflammatory cytokines TNF-α, IL-6, and HYP, and downregulated the expressions of PI3K and Akt mRNA. This study suggests that the anti-HF effect of Wuling Capsules may be achieved by regulating the PI3K-Akt signaling pathway, reducing the levels of TNF-α and IL-6 inflammatory factors, and inhibiting the excessive deposition of collagen.


Asunto(s)
Experimentación Animal , Medicamentos Herbarios Chinos , Animales , Ratas , Interleucina-6 , Farmacología en Red , Factor de Necrosis Tumoral alfa , Simulación del Acoplamiento Molecular , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Cirrosis Hepática/tratamiento farmacológico , Cirrosis Hepática/genética , Medicina Tradicional China , Cápsulas , Fosfatidilinositol 3-Quinasa Clase I , Colágeno , Medicamentos Herbarios Chinos/farmacología
4.
J Anesth ; 29(3): 346-351, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25475993

RESUMEN

PURPOSE: The aim of this study was to investigate the changes in oxidative stress and antioxidants in lung tissue under different tidal volume ventilation conditions. METHODS: Forty-eight male Wistar rats were randomized into four groups, namely, group C, the control group, which was not ventilated, and groups C1, C2 and C3, the treatment groups, which were ventilated for 2 h with tidal volumes of 8, 30 and 42 ml/kg, respectively. The right middle lobe was assayed for malondialdehyde (MDA), the right posterior lobe was assayed using Western blotting for Nrf2, GCLm and SrX1 and the left lobe was assayed for Nrf2, GCLm and SrX1 mRNA. RESULTS: The MDA levels were increased in the three treatment groups, with MDA levels highest in group C3 and lowest in group C1 (C3 > C2 > C1) (all P < 0.05). The mRNA expression of Nrf2, GCLm and SrX1 was highest in group C3 and lowest in group 1 (C3 > C2 > C1) (all P < 0.05). No significant difference was observed between group C1 and group C (P > 0.05). A Western blot analysis showed that Nrf2, GCLm and SrX1 expression was highest in group C3 and lowest in group C1 (C3 > C2 > C1) (all P < 0.05). No significant difference was observed between group C1 and group C (P > 0.05). CONCLUSIONS: Oxidative stress and antioxidant enzyme levels in the lungs of rats were positively associated with the tidal volumes of mechanical ventilation, suggesting that higher tidal volumes cause more severe oxidative stress and increased antioxidant responses.


Asunto(s)
Antioxidantes/metabolismo , Pulmón/metabolismo , Estrés Oxidativo/fisiología , Respiración Artificial/métodos , Animales , Masculino , Malondialdehído/metabolismo , Ratas , Ratas Wistar , Volumen de Ventilación Pulmonar/fisiología
5.
Eur J Pharmacol ; 967: 176383, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38311281

RESUMEN

Toll-like receptor (TLR) 7, a transmembrane signal transduction receptor expressed on the surface of endosomes, has become an attractive target for antiviral and cancer immunotherapies. TLR7 can induce signal transduction by recognizing single-stranded RNA or its analogs, leading to the release of cytokines such as IL-6, IL-12, TNF-α and type-I IFN. Activation of TLR7 helps to enhance immunogenicity and immune memory by stimulating immune cells. Herein, we identified a novel selective TLR7 agonist, GY101, and determined its ability to activate TLR7. In summary, in vitro, compound GY101 significantly induced the secretion of IL-6, IL-12, TNF-α and IFN-γ in mouse splenic lymphocytes; in vivo, peritumoral injection of GY101 significantly suppressed colon cancer CT26, as well as poorly immunogenic B16-F10 and 4T1 cancer cell-derived tumor growth by activating the infiltration of lymphocytes and polarization of M2-like macrophages into M1-like macrophages. These results demonstrate that GY101, as a potent TLR7 agonist, holds great potential for cancer immunotherapy.


Asunto(s)
Neoplasias del Colon , Receptor Toll-Like 7 , Animales , Ratones , Receptor Toll-Like 7/agonistas , Factor de Necrosis Tumoral alfa , Interleucina-6 , Interleucina-12 , Adyuvantes Inmunológicos , Neoplasias del Colon/tratamiento farmacológico
6.
Dev Cell ; 59(3): 384-399.e5, 2024 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-38198890

RESUMEN

Different types of cells uptake fatty acids in response to different stimuli or physiological conditions; however, little is known about context-specific regulation of fatty acid uptake. Here, we show that muscle injury induces fatty acid uptake in muscle stem cells (MuSCs) to promote their proliferation and muscle regeneration. In humans and mice, fatty acids are mobilized after muscle injury. Through CD36, fatty acids function as both fuels and growth signals to promote MuSC proliferation. Mechanistically, injury triggers the translocation of CD36 in MuSCs, which relies on dynamic palmitoylation of STX11. Palmitoylation facilitates the formation of STX11/SNAP23/VAMP4 SANRE complex, which stimulates the fusion of CD36- and STX11-containing vesicles. Restricting fatty acid supply, blocking fatty acid uptake, or inhibiting STX11 palmitoylation attenuates muscle regeneration in mice. Our studies have identified a critical role of fatty acids in muscle regeneration and shed light on context-specific regulation of fatty acid sensing and uptake.


Asunto(s)
Ácidos Grasos , Lipoilación , Músculo Esquelético , Proteínas Qa-SNARE , Regeneración , Animales , Humanos , Ratones , Transporte Biológico , Antígenos CD36/metabolismo , Membrana Celular/metabolismo , Ácidos Grasos/metabolismo , Músculo Esquelético/lesiones , Músculo Esquelético/fisiología , Proteínas Qa-SNARE/metabolismo
7.
Plant Physiol ; 160(2): 696-707, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22885936

RESUMEN

The wall-associated kinase (WAK) gene family is a unique subfamily of receptor-like kinases (RLKs) in plants. WAK-RLKs play roles in cell expansion, pathogen resistance, and metal tolerance in Arabidopsis (Arabidopsis thaliana). Rice (Oryza sativa) has far more WAK-RLK genes than Arabidopsis, but the functions of rice WAK-RLKs are poorly understood. In this study, we found that one rice WAK-RLK gene, DEFECT IN EARLY EMBRYO SAC1 (OsDEES1), is involved in the regulation of early embryo sac development. OsDEES1 silencing by RNA interference caused a high rate of female sterility. Crossing experiments showed that female reproductive organs lacking OsDEES1 carried a functional defect. A detailed investigation of the ovaries from OsDEES1 RNA interference plants indicated that the knockdown of OsDEES1 expression did not affect megasporogenesis but that it disturbed female gametophyte formation, resulting in a degenerated embryo sac and defective seed formation. OsDEES1 exhibited a tissue-specific expression pattern in flowers and seedlings. In the ovary, OsDEES1 was expressed in the megagametophyte region and surrounding nucellus cells in the ovule near the micropylar region. OsDEES1 was found to be a membrane-localized protein with a unique sequence compared with other WAK-RLKs. These data indicate that OsDEES1 plays a role in rice sexual reproduction by regulating female gametophyte development. This study offers new insight into the functions of the WAK-RLK family.


Asunto(s)
Pared Celular/enzimología , Oryza/enzimología , Óvulo Vegetal/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Proteínas Quinasas/metabolismo , Secuencia de Aminoácidos , Membrana Celular/genética , Membrana Celular/metabolismo , Supervivencia Celular , Cruzamientos Genéticos , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Técnicas de Silenciamiento del Gen , Genes de Plantas , Inmunohistoquímica , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Datos de Secuencia Molecular , Oryza/embriología , Oryza/genética , Óvulo Vegetal/enzimología , Infertilidad Vegetal , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/embriología , Plantas Modificadas Genéticamente/enzimología , Plantas Modificadas Genéticamente/genética , Proteínas Quinasas/genética , Interferencia de ARN
8.
Front Plant Sci ; 14: 1100842, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36938012

RESUMEN

Introduction: The decomposition of plant litter mass is responsible for substantial carbon fluxes and remains a key process regulating nutrient cycling in natural and managed ecosystems. Litter decomposition has been addressed in agricultural monoculture systems, but not in intercropping systems, which produce species-diverse litter mass mixtures. The aim here is to quantify how straw type, the soil environment and their combined effects may influence straw decomposition in widely practiced maize/legume intercropping systems. Methods: Three decomposition experiments were conducted over 341 days within a long-term intercropping field experiment which included two nitrogen (N) addition levels (i.e. no-N and N-addition) and five cropping systems (maize, soybean and peanut monocultures and maize/soybean and maize/peanut intercropping). Experiment I was used to quantify litter quality effects on decomposition; five types of straw (maize, soybean, peanut, maize-soybean and maize-peanut) from two N treatments decomposed in the same maize plot. Experiment II addressed soil environment effects on root decomposition; soybean straw decomposed in different plots (five cropping systems and two N levels). Experiment III addressed 'home' decomposition effects whereby litter mass (straw) was remained to decompose in the plot of origin. The contribution of litter and soil effects to the home-field advantages was compared between experiment III ('home' plot) and I-II ('away' plot). Results and discussions: Straw type affected litter mass loss in the same soil environment (experiment I) and the mass loss values of maize, soybean, peanut, maize-soybean, and maize-peanut straw were 59, 77, 87, 76, and 78%, respectively. Straw type also affected decomposition in the 'home' plot environment (experiment III), with mass loss values of maize, soybean, peanut, maize-soybean and maize-peanut straw of 66, 74, 80, 72, and 76%, respectively. Cropping system did not affect the mass loss of soybean straw (experiment II). Nitrogen-addition significantly increased straw mass loss in experiment III. Decomposition of maize-peanut straw mixtures was enhanced more by 'home-field advantage' effects than that of maize-soybean straw mixtures. There was a synergistic mixing effect of maize-peanut and maize-soybean straw mixture decomposition in both 'home' (experiment III) and 'away' plots (experiment I). Maize-peanut showed greater synergistic effects than maize-soybean in straw mixture decomposition in their 'home' plot (experiment III). These findings are discussed in terms of their important implications for the management of species-diverse straw in food-production intercropping systems.

9.
Nat Commun ; 13(1): 6004, 2022 10 12.
Artículo en Inglés | MEDLINE | ID: mdl-36224181

RESUMEN

Aberrant activation of EGFR due to overexpression or mutation is associated with poor prognosis in many types of tumors. Here we show that blocking the sorting system that directs EGFR to plasma membrane is a potent strategy to treat EGFR-dependent tumors. We find that EGFR palmitoylation by DHHC13 is critical for its plasma membrane localization and identify ARF6 as a key factor in this process. N-myristoylated ARF6 recognizes palmitoylated EGFR via lipid-lipid interaction, recruits the exocyst complex to promote EGFR budding from Golgi, and facilitates EGFR transporting to plasma membrane in a GTP-bound form. To evaluate the therapeutic potential of this sorting system, we design a cell-permeable peptide, N-myristoylated GKVL-TAT, and find it effectively disrupts plasma membrane localization of EGFR and significantly inhibits progression of EGFR-dependent tumors. Our findings shed lights on the underlying mechanism of how palmitoylation directs protein sorting and provide an potential strategy to manage EGFR-dependent tumors.


Asunto(s)
Factores de Ribosilacion-ADP , Neoplasias , Factores de Ribosilacion-ADP/metabolismo , Membrana Celular/metabolismo , Receptores ErbB/metabolismo , Guanosina Trifosfato/metabolismo , Humanos , Lípidos , Neoplasias/metabolismo , Transporte de Proteínas
10.
Cryobiology ; 62(2): 135-44, 2011 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-21276438

RESUMEN

Although incubation with glucose before freezing can increase the recovery of human red blood cells frozen with polymer, this method can also result in membrane lesions. This study will evaluate whether addition of oligosaccharide (trehalose, sucrose, maltose, or raffinose) can improve the quality of red blood cell membrane after freezing in the presence of glucose and dextran. Following incubation with glucose or the combinations of glucose and oligosaccharides for 3h in a 37°C water bath, red blood cells were frozen in liquid nitrogen for 24h using 40% dextran (W/V) as the extracellular protective solution. The postthaw quality was assessed by percent hemolysis, osmotic fragility, mean corpuscle volume (MCV), distribution of phosphatidylserine, the postthaw 4°C stability, and the integrity of membrane. The results indicated the loading efficiency of glucose or oligosaccharide was dependent on their concentrations. Moreover, addition of trehalose or sucrose could efficiently decrease osmotic fragility of red blood cells caused by incubation with glucose before freezing. The percentage of damaged cell following incubation with glucose was 38.04±21.68% and significantly more than that of the unfrozen cells (0.95±0.28%, P<0.01). However, with the increase of the concentrations of trehalose, the percentages of damaged cells were decreased steadily. When the concentration of trehalose was 400mM, the percentage of damaged cells was 1.97±0.73% and similar to that of the unfrozen cells (P>0.05). Moreover, similar to trehalose, raffinose can also efficiently prevent the osmotic injury caused by incubation with glucose. The microscopy results also indicated addition of trehalose could efficiently decrease the formation of ghosts caused by incubation with glucose. In addition, the gradient hemolysis study showed addition of oligosaccharide could significantly decrease the osmotic fragility of red blood cells caused by incubation with glucose. After freezing and thawing, when both glucose and trehalose, sucrose, or maltose were on the both sides of membrane, with increase of the concentrations of sugar, the percent hemolysis of frozen red blood cells was firstly decreased and then increased. When the total concentration of sugars was 400mM, the percent hemolysis was significantly less than that of cells frozen in the presence of dextran and in the absence of glucose and various oligosaccharides (P<0.01). However, when both glucose and trehalose were only on the outer side of membrane, with increase of the concentrations of sugars, the percent hemolysis was increased steadily. Furthermore, addition of oligosaccharides can efficiently decrease the osmotic fragility and exposure of phosphatidylserine of red blood cells frozen with glucose and dextran. In addition, trehalose or raffinose can also efficiently mitigate the malignant effect of glucose on the postthaw 4°C stability of red blood cells frozen in the presence of dextran. Finally, addition of trehalose can efficiently protect the integrity of red blood cell membrane following freezing with dextran and glucose. In conclusion, addition of oligosaccharide can efficiently reduce lesions of freezing on red blood cell membrane in the presence of glucose and dextran.


Asunto(s)
Criopreservación/métodos , Crioprotectores/farmacología , Membrana Eritrocítica/efectos de los fármacos , Membrana Eritrocítica/metabolismo , Eritrocitos/efectos de los fármacos , Oligosacáridos/farmacología , Conservación de la Sangre/métodos , Supervivencia Celular/efectos de los fármacos , Dextranos/farmacología , Recuento de Eritrocitos , Eritrocitos/metabolismo , Glucosa/farmacología , Hemólisis/efectos de los fármacos , Humanos , Maltosa/farmacología , Fragilidad Osmótica/efectos de los fármacos , Fosfatidilserinas/sangre , Fosfatidilserinas/metabolismo , Rafinosa/farmacología , Sacarosa/farmacología , Temperatura , Trehalosa/farmacología
11.
Environ Sci Pollut Res Int ; 28(9): 11618-11636, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33128145

RESUMEN

The thermal effect on the flow and dispersion of pollutants emitted from a rooftop stack is investigated by means of CFD (computational fluid dynamics) models with wind tunnel experimental validations. The leeward wall and its nearby ground are heated simultaneously to mimic solar radiation. Seventeen Ri (Richardson number) cases with four inflow wind speeds (1, 3, 6, and 9 m/s) and five temperature differences (0, 60, 120, 180, and 240 K) between the heated surface and ambient air are considered to represent the interaction between thermal buoyancy force and inertia force. The results reveal that (1) the steady RANS (Reynolds Averaged Navier-Stokes) computations with Boussinesq approximation can generally reproduce the effect of thermal buoyancy on the wake flow and pollutant distribution in wind tunnel experiments; (2) the wake vortex flow is less affected by the thermal buoyancy force at small Ri (e.g., Ri ≤ 0.26) while an upward flow rather than a clockwise vortex structure is developed in the near wake at Ri ≥ 0.58; (3) it is inappropriate to place fresh air intakes on the leeward wall of the emitting building, but natural ventilation through windows on the leeward wall can be implemented at higher Ri (e.g., Ri = 2.33); (4) at the pedestrian respiration height downstream of the building, the distance between the location of maximum pollutant concentration and the leeward wall increases linearly with Ri while the maximum dimensionless concentration decreases exponentially with increasing Ri; (5) the air temperature is rapidly reduced away from the heated wall/ground and a heat accumulation zone is formed at the ground corner next to the leeward wall. This study can be helpful for determining the strategy for natural ventilation through windows and for evaluating the impacts of rooftop stack exhaust on air quality downstream of emitting buildings.


Asunto(s)
Contaminantes Atmosféricos , Contaminación del Aire , Contaminantes Atmosféricos/análisis , Hidrodinámica , Modelos Teóricos , Temperatura
12.
Front Physiol ; 12: 777908, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34955890

RESUMEN

The Asian citrus psyllid (ACP) Diaphorina citri is the main vector of the pathogen Candidatus Liberibacter asiaticus (CLas), which is the causal agent of citrus Huanglongbing disease. Feeding by both ACP nymphs and adults on host plants allows them to obtain nutrition. Therefore, the nutritional content within the plant phloem is of much importance for the development and reproduction of ACP. The infection by pathogenic microbiomes may affect the amino acid contents of their host plants and then indirectly affect the biology of sap-feeding insects. In this study, we investigated the amino acid contents and their proportions in both CLas-infected and CLas-free citrus plants, ACP adults, and also in honeydew produced by ACP nymphs. Results showed that infection by CLas had a large impact on the amino acid species and proportion in all the tested target plants, ACP adults, and in the honeydew of ACP nymphs. The content of total amino acids in CLas-infected citrus was much higher than that of CLas-free citrus. However, CLas infection significantly reduced the proportion of essential amino acids (EAAs) in these plants. When feeding on CLas-infected citrus plants, ACP adults absorbed less total amino acids than those adults feeding on healthy plants, but the proportion of EAAs was significantly higher when they fed on CLas-infected citrus plants. The proportion of EAAs also significantly increased in the honeydew secreted by ACP nymphs that fed on CLas-infected citrus plants. However, EAA detection in the honeydew of ACP nymphs indicated that the utilization rate of EAAs by CLas positive ACP nymphs was reduced. Our study has revealed that CLas infection significantly affects the contents, proportion, and utilization efficiency of different amino acids in citrus plants, ACP adults, and nymphs, leading to a developmental pattern of ACP that is more conducive to CLas transmission.

13.
Insect Sci ; 27(5): 1053-1066, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31454147

RESUMEN

Alcohol dehydrogenase 5 (ADH5) is a member of medium-chain dehydrogenase/reductase family and takes part in cellular formaldehyde and S-nitrosoglutathione metabolic network. 2-tridecanone (2-TD) is a toxic compound in many Solanaceae crops to defend against a variety of herbivory insects. In the broader context of insect development and pest control strategies, this study investigates how a new ADH5 from Helicoverpa armigera (HaADH5) regulates the expression of CYP6B6, a gene involved in molting and metamorphosis, in response to 2-TD treatment. Cloning of the HaADH5 complementary DNA sequence revealed that its 1002 bp open reading frame encodes 334 amino acids with a predicted molecular weight of 36.5 kD. HaADH5 protein was purified in the Escherichia coli Transetta (pET32a-HaADH5) strain using a prokaryotic expression system. The ability of HaADH5 protein to interact with the 2-TD responsive region within the promoter of CYP6B6 was confirmed by an in vitro electrophoretic mobility shift assay and transcription activity validation in yeast. Finally, the expression levels of both HaADH5 and CYP6B6 were found to be significantly decreased in the midgut of 6th instar larvae after 48 h of treatment with 10 mg/g 2-TD artificial diet. These results indicate that upon 2-TD treatment of cotton bollworm, HaADH5 regulates the expression of CYP6B6 by interacting with its promoter. As HaADH5 regulation of CYP6B6 expression may contribute to the larval xenobiotic detoxification, molting and metamorphosis, HaADH5 is a candidate target for controlling the growth and development of cotton bollworm.


Asunto(s)
Aldehído Oxidorreductasas/genética , Familia 6 del Citocromo P450/genética , Proteínas de Insectos/genética , Cetonas/metabolismo , Mariposas Nocturnas/genética , Aldehído Oxidorreductasas/química , Aldehído Oxidorreductasas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Familia 6 del Citocromo P450/química , Familia 6 del Citocromo P450/metabolismo , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Larva/enzimología , Larva/crecimiento & desarrollo , Larva/metabolismo , Mariposas Nocturnas/enzimología , Mariposas Nocturnas/crecimiento & desarrollo , Filogenia
14.
Microbiologyopen ; 7(3): e00561, 2018 06.
Artículo en Inglés | MEDLINE | ID: mdl-29573202

RESUMEN

Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed.


Asunto(s)
Hemípteros/crecimiento & desarrollo , Hemípteros/microbiología , Insectos Vectores/microbiología , Wolbachia/aislamiento & purificación , Estructuras Animales/microbiología , Animales , Asia , Carga Bacteriana , Femenino , Hibridación Fluorescente in Situ , Masculino , Nymphaea/microbiología , Cigoto/microbiología
15.
Zhonghua Nan Ke Xue ; 12(9): 798-9, 802, 2006 Sep.
Artículo en Zh | MEDLINE | ID: mdl-17009530

RESUMEN

OBJECTIVE: To investigate the characteristics of erectile dysfunction (ED) in old males with lacunar infarction. METHODS: A total of 38 old patients ages from 60 to 70 years were involved. The questionnaire of international index of erectile function 5 (IIEF -5) was used to determine the status and severity of ED. According to the focus of infarction on MRI, the patients were divided into two groups, Group I with lacunar infarction and minor neurological deficits, and Group II with none. The total IIEF-5 scores were compared between the two groups and repeatedly evaluated six months after discharge. RESULTS: According to the total scores of IIEF-5, the prevalence of ED in Group II (95%) was higher, and the incidence of severe ED was significantly increased (60.0% vs. 44.4%, P < 0.05) as compared with Group II. In both the two groups, severe ED was more often seen in diabetic patients. At six months after discharge, the total scores of IIEF-5 were significantly increased (11.2 +/- 3.2 vs. 15.6 +/- 2.2, P < 0.05). CONCLUSION: ED is significantly increased in old males with lacunar infarction, and it is more severe in diabetic patients. Post-stroke rehabilitation care helps to improve ED.


Asunto(s)
Infarto Encefálico/complicaciones , Disfunción Eréctil/epidemiología , Erección Peniana , Anciano , Infarto Encefálico/rehabilitación , Complicaciones de la Diabetes , Disfunción Eréctil/etiología , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Encuestas y Cuestionarios
16.
Guang Pu Xue Yu Guang Pu Fen Xi ; 25(6): 975-8, 2005 Jun.
Artículo en Zh | MEDLINE | ID: mdl-16201387

RESUMEN

Four new complexes of Ni(II), Cu(II), Zn(II) and Co(III) with HL (HL= N,N'-di-beta-D-glucosylethylenediamine) have been synthesized in methanol solutions. The compositions of the complexes determined by elementary analysis are [Ni(HL)(H2O)2]2+ Cl2.CH3OH.2H2O, [Cu(HL)]2+ Cl2.CH3CH2OH.3H2O, [Zn(HL)]2+ Cl2.H2O and [Co(HL)(H2O)(OH)]2+ Cl2.CH3OH.2H2O respectively. They were also characterized by IR, UV and NMR spectra which show that complexes of Ni(II) and Co(III) have octahedral configuration, while complexes of Cu(II) and Zn(II) have tetralhedral configuration. The hydrolytic kinetics of p-Nitrophenyl picolinate(PNPP) catalyzed by these complexes in buffer solution was also studied spectrophotometrically.


Asunto(s)
Glicósidos/química , Metales/química , Compuestos Organometálicos/química , Análisis Espectral/métodos , Catálisis , Cobalto/química , Cobre/química , Ésteres , Glicósidos/síntesis química , Glicósidos/metabolismo , Hidrólisis , Cinética , Espectroscopía de Resonancia Magnética , Modelos Químicos , Estructura Molecular , Níquel/química , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/metabolismo , Ácidos Picolínicos/química , Ácidos Picolínicos/metabolismo , Espectrofotometría Infrarroja , Espectrofotometría Ultravioleta , Zinc/química
17.
Medicine (Baltimore) ; 94(6): e502, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25674742

RESUMEN

Our previous study indicated overexpression of metadherin (MTDH) is an adverse prognostic factor in squamous cell carcinoma of the head and neck (SCCHN) and promotes SCCHN cell proliferation and invasion. However, its mechanism remains unclear. Recent studies have indicated that MTDH is a cancer-metastasis-associated molecule that participates in the process of angiogenesis. Therefore, the study is aimed to investigate that whether vascular endothelial growth factor (VEGF), as one of the most potent proangiogenic cytokines, is regulated by MTDH and the role of the phosphatidylinositide 3-kinases/Protein Kinase B (PI3K/Akt) pathway in this process of regulation and the clinical significance of both MTDH and VEGF in SCCHN.Immunohistochemistry was used to assay the expression of MTDH and VEGF in a cohort of 189 SCCHN patients with intact follow-up information. The expression of MTDH was then upregulated or inhibited by lentivirus-mediated MTDH Complementary deoxyribonucleic acid or MTDH short hairpin ribonucleic acid (shRNA) to observe the resulting alterations in VEGF expression and the PI3K/Akt signaling pathway in SCCHN cell lines. In addition, the PI3K/Akt pathway was modulated to observe the resulting changes in the MTDH-mediated expression of VEGF.The immunohistochemistry data showed that MTDH expression is positively correlated with VEGF expression in SCCHN tissues. Moreover, the overexpression of MTDH in SCCHN Tu686 and 5-8F cells led to increases in the expression of VEGF, and this effect was accompanied by activation of the PI3K/Akt pathway. Conversely, shRNA-mediated knockdown of MTDH led to decreased VEGF expression. In addition, inhibition of the Akt signaling pathway reversed the upregulation of VEGF resulting from MTDH overexpression. Moreover, the survival analysis revealed that VEGF is an independent prognostic factor, and a combined survival analysis based on both MTDH and VEGF showed synergistic effects in the prognosis evaluation of SCCHN patients.The findings of the present study demonstrate that MTDH regulates the expression of VEGF via the PI3K/Akt signaling pathway, indicating the potential role of the MTDH-mediated activation of VEGF signaling pathway in SCCHN angiogenesis and metastasis.


Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Moléculas de Adhesión Celular/fisiología , Neoplasias de Cabeza y Cuello/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Western Blotting , Carcinoma de Células Escamosas/mortalidad , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Neoplasias de Cabeza y Cuello/mortalidad , Humanos , Inmunohistoquímica , Proteínas de la Membrana , Pronóstico , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , ARN Interferente Pequeño/fisiología , Proteínas de Unión al ARN , Transducción de Señal/fisiología , Transfección , Regulación hacia Arriba
18.
Burns ; 41(8): 1748-1757, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26514700

RESUMEN

BACKGROUND: Puerarin, the main isoflavone glycoside extracted from the root of Pueraria lobata, is widely prescribed for patients with cardiovascular disorders in China. This study investigates the effect of puerarin on severe burn-induced acute myocardial injury in rats and its underlying mechanisms. MATERIALS AND METHODS: Healthy adult Wistar rats were divided into three groups: (1) sham group, sham burn treatment; (2) burn group, third-degree burns over 30% of the total body surface area (TBSA) with lactated Ringer's solution for resuscitation; and (3) burn plus puerarin group, third-degree burns over 30% of TBSA with lactated Ringer's solution containing puerarin for resuscitation. The burned animals were sacrificed at 1, 3, 6, 12, and 24 h after burn injury. Myocardial injury was evaluated by analyzing serum creatine kinase MB fraction (CK-MB) activity and cardiac troponin T (cTNT) level. Changes in cardiomyocyte ultrastructure were also determined using a transmission electron microscope. Tumor necrosis factor (TNF)-α concentration in serum was measured by radioimmunoassay. Cardiac myeloperoxidase (MPO) activity and malondialdehyde (MDA) concentration were measured to determine neutrophil infiltration and oxidative stress in the heart, respectively. The expression of p38 mitogen-activated protein (MAP) kinase in the heart was determined by Western blot analysis. RESULTS: After the 30% TBSA full-thickness burn injury, serum CK-MB activities and cTnT levels increased markedly, both of which were significantly decreased by the puerarin treatment. The level of serum TNF-α concentration in burn group at each time-point was obviously higher than those in sham group (1.09±0.09 ng/ml), and it reached the peak value at 12 h post burn. Burn trauma also resulted in worsen ultrastructural condition, elevated MPO activity and MDA content in heart tissue, and a significant activation of cardiac p38 MAP kinase. Administration of puerarin improved the ultrastructural changes in cardiomyocytes, decreased TNF-α concentration in serum as well as suppressed cardiac MPO activity and reduced MDA content, and abolished the activation of p38 MAP kinase in heart tissue after severe burn. CONCLUSIONS: These results suggest that puerarin attenuates inflammatory responses, reduces neutrophil infiltration and oxidative stress in the heart, and protects against acute myocardial injury induced by severe burn.


Asunto(s)
Quemaduras/complicaciones , Corazón/efectos de los fármacos , Isoflavonas/farmacología , Isquemia Miocárdica/etiología , Miocardio/inmunología , Infiltración Neutrófila/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Vasodilatadores/farmacología , Animales , Forma MB de la Creatina-Quinasa/sangre , Forma MB de la Creatina-Quinasa/efectos de los fármacos , Inflamación , Malondialdehído/metabolismo , Isquemia Miocárdica/sangre , Miocardio/metabolismo , Peroxidasa/efectos de los fármacos , Peroxidasa/metabolismo , Ratas , Ratas Wistar , Índices de Gravedad del Trauma , Troponina T/sangre , Troponina T/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
19.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 25(3): 267-70, 2003 Jun.
Artículo en Zh | MEDLINE | ID: mdl-12905737

RESUMEN

OBJECTIVE: To measure hip bone mineral density (BMD) on different femoral rotations and to identify the influence of femoral rotation during hip densitometry. METHODS: Forty women were recruited at the out-department of orthopedic, with the average age of 60.7 years (45-85 years), the average height of 161.8 cm (150-175 cm), and the average weight of 67.4 kg (48-80 kg). BMD measurement of hip was done in neutral position (0 degrees), 15 degrees and 30 degrees of internal rotation under quality control of DXA machine. All the subjects were asked to get down scan table and reposition after each measuring in different rotation. RESULTS: Hip BMD of proximal femur (femoral neck, Ward's area and trochanter region) have significant variations in measurement (P < 0.01) among neutral position, 15 degrees and 30 degrees of internal rotation. There were more BMD change at the cancellous region (Ward's triangle and trochanter region) than at cortical bone area (femoral neck region). Statistical analysis showed a significant positive correlation for hip BMD change between neutral degrees to 15 degrees of internal rotation and 15 degrees to 30 degrees of internal rotation. But no correlation between neutral degrees to 30 degrees of internal rotation for hip BMD change. CONCLUSIONS: Femoral rotation was shown to have a significant effect on hip BMD measurements. Cortical bone region of hip have a less BMD change than cancellous bone region. Proper positioning of the femur during scan can improve precision significantly.


Asunto(s)
Absorciometría de Fotón/métodos , Densidad Ósea , Fémur , Rotación , Anciano , Anciano de 80 o más Años , Femenino , Cadera , Humanos , Persona de Mediana Edad , Postura
20.
Pest Manag Sci ; 70(10): 1604-10, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24488542

RESUMEN

BACKGROUND: Whitefly Bemisia tabaci is a globally distributed and most destructive pest to agriculture. Owing to increasing chemical resistance, a long-lasting strategy to manage this pest must involve biological control. Herbivore-induced plant volatiles (HIPVs) usually play a profoundly important role in the foraging behaviour of natural enemies. Here, the effects of HIPVs from Chinese broccoli on the foraging behaviour of two dominant natural enemy species of B. tabaci in China, Encarsia formosa and Serangium japonicum, were investigated using a four-arm olfactometer, and exogenous jasmonic acid (JA) was used to induce plant volatiles to mimic the damage of the herbivore pest. RESULTS: The parasitoid E. formosa was found to be more attracted by the volatiles from JA-induced broccoli than those from control plants. The residence times of E. formosa in the final-choice areas closed to volatiles from shoot JA (SJA)- and root JA (RJA)-induced plants were 119.8 ± 35.2 s and 99.8 ± 34.7 s respectively in the dual-choice experiments, and 123.8 ± 32.0 s and 102.3 ± 28.7 s respectively in the three-choice experiment. All are significantly longer than those spent in the final-choice area closed to volatiles from control plants (CON). JA-induced volatiles were also only a little more attractive to the predator S. japonicum. However, there was no significant difference between each of the two natural enemies in residence time spent in the final-choice areas closed to SJA, RJA or CON volatiles in both the dual- and three-choice experiments. Furthermore, the number of times that the parasitoid or predator entered the SJA, RJA and CON final-choice areas was not significantly different in any of the experiments. CONCLUSION: The present results indicate that RJA- and SJA-induced plant volatiles have higher efficiencies in attracting the natural enemies of whitefly B. tabaci. The attractive efficiencies varied according to the plant volatiles that JA induced, and also depended on the natural enemy species. The parasitoid E. formosa seems to be more sensitive to and attracted by the induced volatiles than the predator S. japonicum.


Asunto(s)
Brassica/química , Hemípteros , Control Biológico de Vectores/métodos , Reguladores del Crecimiento de las Plantas/fisiología , Animales , Brassica/parasitología , China , Escarabajos/fisiología , Ciclopentanos , Femenino , Herbivoria , Himenópteros/fisiología , Odorantes , Oxilipinas
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