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1.
Front Nutr ; 11: 1335538, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38562486

RESUMEN

The Chinese name "Lingzhi" refers to Ganoderma genus, which are increasingly used in the food and medical industries. Ganoderma species are often used interchangeably since the differences in their composition are not known. To find compositional metabolite differences among Ganoderma species, we conducted a widely targeted metabolomics analysis of four commonly used edible and medicinal Ganoderma species based on ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Through pairwise comparisons, we identified 575-764 significant differential metabolites among the species, most of which exhibited large fold differences. We screened and analyzed the composition and functionality of the advantageous metabolites in each species. Ganoderma lingzhi advantageous metabolites were mostly related to amino acids and derivatives, as well as terpenes, G. sinense to terpenes, and G. leucocontextum and G. tsugae to nucleotides and derivatives, alkaloids, and lipids. Network pharmacological analysis showed that SRC, GAPDH, TNF, and AKT1 were the key targets of high-degree advantage metabolites among the four Ganoderma species. Analysis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes demonstrated that the advantage metabolites in the four Ganoderma species may regulate and participate in signaling pathways associated with diverse cancers, Alzheimer's disease, and diabetes. Our findings contribute to more targeted development of Ganoderma products in the food and medical industries.

2.
Zool Res ; 45(1): 189-200, 2024 Jan 18.
Artículo en Inglés | MEDLINE | ID: mdl-38199973

RESUMEN

Monitoring the prevalence of antimicrobial resistance genes (ARGs) is vital for addressing the global crisis of antibiotic-resistant bacterial infections. Despite its importance, the characterization of ARGs and microbiome structures, as well as the identification of indicators for routine ARG monitoring in pig farms, are still lacking, particularly concerning variations in antimicrobial exposure in different countries or regions. Here, metagenomics and random forest machine learning were used to elucidate the ARG profiles, microbiome structures, and ARG contamination indicators in pig manure under different antimicrobial pressures between China and Europe. Results showed that Chinese pigs exposed to high-level antimicrobials exhibited higher total and plasmid-mediated ARG abundances compared to those in European pigs ( P<0.05). ANT(6)-Ib, APH(3')-IIIa, and tet(40) were identified as shared core ARGs between the two pig populations. Furthermore, the core ARGs identified in pig populations were correlated with those found in human populations within the same geographical regions. Lactobacillus and Prevotella were identified as the dominant genera in the core microbiomes of Chinese and European pigs, respectively. Forty ARG markers and 43 biomarkers were able to differentiate between the Chinese and European pig manure samples with accuracies of 100% and 98.7%, respectively. Indicators for assessing ARG contamination in Chinese and European pigs also achieved high accuracy ( r=0.72-0.88). Escherichia flexneri in both Chinese and European pig populations carried between 21 and 37 ARGs. The results of this study emphasize the importance of global collaboration in reducing antimicrobial resistance risk and provide validated indicators for evaluating the risk of ARG contamination in pig farms.


Asunto(s)
Antiinfecciosos , Microbioma Gastrointestinal , Humanos , Animales , Porcinos , Antibacterianos/farmacología , Estiércol , Farmacorresistencia Bacteriana/genética
3.
Sci Total Environ ; 882: 163511, 2023 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-37080303

RESUMEN

Integrated and monoculture freshwater aquaculture systems are often regarded as important reservoirs for antimicrobial resistance genes (ARGs) and antimicrobial resistance bacteria (ARBs), yet only a few studies have assessed differences in the antimicrobial resistome and antibiotic residues between aquaculture modes. In this study, a metagenomic approach was used to comprehensively explore the dynamic patterns and potential transmission mechanisms of ARGs in ducks, human workers, fish, water and sediments during the transition from an integrated to a monoculture freshwater aquaculture mode and to investigate the associations of ARGs with potential hosts in microbial communities using network analysis and a binning approach. The results showed that the abundance and diversity of ARGs were higher under integrated fish-duck farming than in single fish ponds. During the transition from an integrated to a monoculture aquaculture farm, ARGs in workers and sediments were not easily removed. However, ARGs in the aquatic environment underwent regular changes. In addition, duck manure was probably the most dominant source of ARGs in the duck farm environment. Network analysis indicated that Escherichia spp. were the most dominant hosts of ARGs. Variation partitioning analysis (VPA) showed that in water samples, the bacterial community played an important role in the ARG profile. In addition, we identified a potential risk of the presence of highly virulent and antimicrobial-resistant Klebsiella pneumoniae in workers. These results help assess the risk of ARG transmission in integrated and monoculture aquaculture farms and suggest that we should strengthen the monitoring of long-term resistance in integrated aquaculture environments.


Asunto(s)
Antagonistas de Receptores de Angiotensina , Genes Bacterianos , Animales , Humanos , Antagonistas de Receptores de Angiotensina/análisis , Farmacorresistencia Microbiana/genética , Inhibidores de la Enzima Convertidora de Angiotensina/análisis , Bacterias/genética , Acuicultura/métodos , Antibacterianos/análisis , Agua/análisis , China
4.
Neuroscience Bulletin ; (6): 245-260, 2023.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-971567

RESUMEN

Defensive behaviors induced by innate fear or Pavlovian fear conditioning are crucial for animals to avoid threats and ensure survival. The zona incerta (ZI) has been demonstrated to play important roles in fear learning and fear memory, as well as modulating auditory-induced innate defensive behavior. However, whether the neuronal subtypes in the ZI and specific circuits can mediate the innate fear response is largely unknown. Here, we found that somatostatin (SST)-positive neurons in the rostral ZI of mice were activated by a visual innate fear stimulus. Optogenetic inhibition of SST-positive neurons in the rostral ZI resulted in reduced flight responses to an overhead looming stimulus. Optogenetic activation of SST-positive neurons in the rostral ZI induced fear-like defensive behavior including increased immobility and bradycardia. In addition, we demonstrated that manipulation of the GABAergic projections from SST-positive neurons in the rostral ZI to the downstream nucleus reuniens (Re) mediated fear-like defensive behavior. Retrograde trans-synaptic tracing also revealed looming stimulus-activated neurons in the superior colliculus (SC) that projected to the Re-projecting SST-positive neurons in the rostral ZI (SC-ZIrSST-Re pathway). Together, our study elucidates the function of SST-positive neurons in the rostral ZI and the SC-ZIrSST-Re tri-synaptic circuit in mediating the innate fear response.


Asunto(s)
Ratones , Animales , Zona Incerta/metabolismo , Neuronas/metabolismo , Miedo/fisiología , Somatostatina/metabolismo
5.
Front Microbiol ; 13: 903983, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36171750

RESUMEN

Artificial cultivation of Morchella sextelata and other morels is expanding in China, but continuous cropping reduces Morchella for unknown reasons. Here, we investigated soil that had been used or not used for M. sextelata cultivation for 0, 1, and 2 years. We found that the continuous cropping of M. sextelata substantially reduced the pH and the nutrient content of the hyphosphere soil and increased sclerotia formation by M. sextelata. Changes in the structure of bacterial and fungal communities were associated with levels of available nitrogen (N) and phosphorus in the soil. With continuous cropping, the richness and diversity of fungal and bacterial communities increased, but the abundance of Bacillus and Lactobacillus decreased and the abundance of pathogenic fungi increased. FAPROTAX analysis indicated that N cycle functions were enriched more with than without continuous cultivation, and that enrichment of N cycle and sulfate respiration functions was higher in the second than in the first year of cultivation. FunGuild analysis indicated that the functions related to pathotrophs and wood saprotrophs were enriched by M. sextelata cultivation. Overall, the results suggest that continuous cropping may reduce M. sextelata production by acidifying the soil and increasing the abundance of pathogenic fungi. Additional research is needed to determine whether increases in the abundance of pathogenic fungi and changes in soil chemistry result in the declines in production that occur with continuous M. sextelata cultivation.

6.
Neuroscience Bulletin ; (6): 565-575, 2022.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-929107

RESUMEN

Licking behavior is important for water intake. The deep mesencephalic nucleus (DpMe) has been implicated in instinctive behaviors. However, whether the DpMe is involved in licking behavior and the precise neural circuit behind this behavior remains unknown. Here, we found that the activity of the DpMe decreased during water intake. Inhibition of vesicular glutamate transporter 2-positive (VGLUT2+) neurons in the DpMe resulted in increased water intake. Somatostatin-expressing (SST+), but not protein kinase C-δ-expressing (PKC-δ+), GABAergic neurons in the central amygdala (CeA) preferentially innervated DpMe VGLUT2+ neurons. The SST+ neurons in the CeA projecting to the DpMe were activated at the onset of licking behavior. Activation of these CeA SST+ GABAergic neurons, but not PKC-δ+ GABAergic neurons, projecting to the DpMe was sufficient to induce licking behavior and promote water intake. These findings redefine the roles of the DpMe and reveal a novel CeASST-DpMeVGLUT2 circuit that regulates licking behavior and promotes water intake.

7.
Chinese Journal of Pediatrics ; (12): 413-420, 2022.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-935713

RESUMEN

Objective: To explore current vitamin D status and influential factors of vitamin D deficiency and insufficiency among children under 7 years of age in 11 provinces, autonomous regions or municipalities of China. Methods: According to the "province-city-hospital" sampling technical route, a total of 1 531 healthy children under 7 years of age were sampled from 11 provinces, autonomous regions or municipalities in China by the cluster random sampling method from November 2020 to November 2021. The demographic information, family conditions, behavior and living habits and feeding behaviors were collected using unified questionnaire. Serum 25-hydroxyvitamin D(25(OH)D) levels were measured by liquid chromatography-tandem mass spectrometry. Serum 25(OH)D<30 nmol/L was considered deficient and 30-50 nmol/L was considered insufficient. With 25(OH)D≤50 nmol/L as the dependent variable, multivariate Logistic regression was applied to analyze the association between vitamin D deficiency and insufficiency and potential influential factors. Results: The prevalence of vitamin D deficiency and insufficiency among children under 7 years of age in 11 provinces, autonomous regions or municipalities of China was 14.0% (215/1 531), 3.8% (25/664) and 21.9% (190/867) in 0-<3 and 3-<7 of age years, respectively. Compared to children aged 0-<3 years, children aged 3-<7 years had a 2.6-fold increased risk of vitamin D deficiency and insufficiency (OR=3.60, 95%CI 1.93-6.72, P<0.001). Frequent sunlight exposure (OR=0.46, 95%CI 0.29-0.73, P=0.001), vitamin D supplementation (sometimes, OR=0.33, 95%CI 0.21-0.51, P<0.001; daily, OR=0.20, 95%CI 0.11-0.36, P<0.001) and infant formula intake(4-7 times per weeks, OR=0.43, 95%CI 0.28-0.68, P<0.001) were protective factors for vitamin D deficiency and insufficiency. Conclusion: Vitamin D deficiency and insufficiency are common among children under 7 years of age in 11 provinces, autonomous regions or municipalities of China, which is affected by age, sunlight exposure, vitamin D supplementation and infant formula intake.


Asunto(s)
Niño , Humanos , Lactante , China/epidemiología , Estudios Transversales , Vitamina D , Deficiencia de Vitamina D/epidemiología , Vitaminas
8.
Chinese Journal of Epidemiology ; (12): 1139-1141, 2013.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-321705

RESUMEN

In many studies about biomedical research factors influence on the outcome variable,it has no influence or has a positive effect within a certain range.Exceeding a certain threshold value,the size of the effect and/or orientation will change,which called threshold effect.Whether there are threshold effects in the analysis of factors (x) on the outcome variable (y),it can be observed through a smooth curve fitting to see whether there is a piecewise linear relationship.And then using segmented regression model,LRT test and Bootstrap resampling method to analyze the threshold effect.Empower Stats software developed by American X & Y Solutions Inc has a threshold effect analysis module.You can input the threshold value at a given threshold segmentation simulated data.You may not input the threshold,but determined the optimal threshold analog data by the software automatically,and calculated the threshold confidence intervals.

9.
Chinese Journal of Epidemiology ; (12): 1173-1175, 2013.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-321698

RESUMEN

Objective The objective of this study was to analyze the related factors that influencing the level of urinary iodine (UI).Methods 994 adult cases were selected from medical centers.Morning urine was collected and questionnaire including age,sex,family medical history of thyroid disease administered.Information on previous 24-hour consumption of iodine-containing foods was collected.Data was analyzed by Empower(R) software with logistic model.Results The median UI level was 193.0 μg/L.After adjusting for potential confounders,gender (P=0.000),family monthly income per capita (P=0.000),the amount of iodized salt intake (P =0.041),and eating kelps (P=0.000) appeared to be associated with the level of UI.Risk regarding the prevalence of excess UI (> 300 μg/L) increased with the increasing amount of salt and kelp consumption:salt consumption >165 g/m (OR=24.3,95%CI:1.1-523.8,P<0.05) ; kelp consumption (OR=9.6,95%CI:2.6-35.1,P<0.001).Conclusion UI was associated with factors as:gender,family monthly income per capita,intake of iodized salt and the amount of kelp consumption.Excessive intake of iodine might be associated with high intake of iodized salt and kelp.

10.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-314978

RESUMEN

<p><b>OBJECTIVE</b>To clone SABATH methyltransferase (rLjSABATHMT) gene in Lonicera japonica var. chinensis, and compare the gene expression and intron sequence of SABATH methyltransferase orthologous in L. japonica with L. japonica var. chinensis. It provide a basis for gene regulate the formation of L. japonica floral scents.</p><p><b>METHOD</b>The cDNA and genome sequences of LjSABATHMT from L. japonica var. chinensis were cloned according to the gene fragments in cDNA library. The LjSABATHMT protein was characterized by bioinformatics analysis. SABATH family phylogenetic tree were built by MEGA 5.0. The transcripted level of SABATHMT orthologous were analyzed in different organs and different flower periods of L. japonica and L. japonica var. chinensis using RT-PCR analysis. Intron sequences of SABATHMT orthologous were also analyzied.</p><p><b>RESULT</b>The cDNA of LjSABATHMT was 1 251 bp, had a complete coding frame with 365 amino acids. The protein had the conservative SABATHMT domain, and phylogenetic tree showed that it may be a salicylic acid/benzoic acid methyltransferase. Higher expression of SABATH methyltransferase orthologous was found in flower. The intron sequence of L. japonica and L. japonica var. chinensis had rich polymorphism, and two SNP are unique genotype of L. japonica var. chinensis. The motif elements in two orthologous genes were significant differences.</p><p><b>CONCLUSION</b>The intron difference of SABATH methyltransferase orthologous could be inducing to difference of gene expression between L. japonica and L. japonica var. chinensis. These results will provide important base on regulating active compounds of L. japonica.</p>


Asunto(s)
Secuencia de Bases , Clonación Molecular , Biología Computacional , Evolución Molecular , Regulación de la Expresión Génica de las Plantas , Lonicera , Genética , Metiltransferasas , Genética , Motivos de Nucleótidos , Filogenia , Alineación de Secuencia , Análisis de Secuencia
11.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-266089

RESUMEN

<p><b>OBJECTIVE</b>To explore the exposure level and risk factors of heavy metal among Shanghai infants in their prenatal period.</p><p><b>METHODS</b>A total of 1652 pregnant or puerperal women were recruited from 10 midwifery-qualified hospitals in Shanghai since October 2008 to October 2009, by the stratified cluster sampling method. They answered the questionnaire and their umbilical cord blood and serum were collected to detect the content of Pb, Hg, Mn, Cd, As and Tl. The risk factors were analyzed by single and multiple regression methods respectively.</p><p><b>RESULTS</b>The median value of the content of Pb, Hg, Mn, Cd, As and Tl were 41.00, 1.88, 4.10, 0.03, 0.86 and 0.02 µg/L respectively. The Hg concentration of pregnant women who ate fish for 4 - 7 times per week (9.7% (160/1652)) was 2.76 µg/L, which was higher than that of pregnant women who only ate fish for 1-3 times per week (49.3% (814/1652)) and those who seldom ate fish (40.0% (661/1652)); the Hg concentration in the two groups above were 2.41 and 2.03 µg/L separately. The difference had statistical significance (χ(2) = 36.40, P < 0.001). Meanwhile, the concentrations of Pb and Tl in the group of pregnant women whose houses were remodeled by PVC pipe (85.0% (1404/1652)) was higher than the concentrations in group of pregnant women whose houses were remodeled by galvanized pipe (15.0% (248/1652)); the Pb concentration in the two groups above were 45.54 and 40.00 µg/L (Z = 2.54, P < 0.05) and the Tl concentration in the two groups above were 0.021 and 0.018 µg/L (Z = 2.97, P < 0.05). However, the As concentration in the group of PVC pipe remodeled was 4.33 µg/L, which was lower than that in the group of galvanized pipe (9.37 µg/L). The difference had statistical significance (Z = 3.99, P < 0.01). The concentrations of Mn, Cd and Tl in the groups of pregnant women whose house had been remodeled in the last 3 years (38.0% (628/1652)) were 14.78, 0.51 and 0.022 µg/L separately, which were all significantly higher than those in the groups of women whose houses' were not remodeled (62.0% (1024/1652)), whose concentrations were 11.01, 0.29 and 0.02 µg/L respectively. The differences had statistical significance (Mn: Z = 2.46, P < 0.05; Cd: Z = 2.38, P < 0.05; Tl: Z = 2.81, P < 0.01).</p><p><b>CONCLUSION</b>The infants in Shanghai were exposed to heavy metals in their prenatal period. The main sources of the exposure were daily diet and remodeled pollution.</p>


Asunto(s)
Adulto , Femenino , Humanos , Recién Nacido , Masculino , Embarazo , China , Contaminación Ambiental , Sangre Fetal , Química , Exposición Materna , Metales Pesados , Factores de Riesgo , Encuestas y Cuestionarios
12.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-322807

RESUMEN

<p><b>OBJECTIVE</b>To explore the effect of trichostatin A (TSA) on human lung cancer cell strains A549.</p><p><b>METHODS</b>A549 cells were exposed to TSA at different concentrations, then the growth-inhibiting effects of the cell line were detected with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay; After the cells were exposed to TSA for 48 and 96 hours at 300 nmol/L, the change of the cell cycle and apoptosis of A549 were analyzed with flow cytometry. p21 protein and extracellular signal regulated kinase (ERK) expression were detected by Western blot.</p><p><b>RESULTS</b>TSA inhibited the growth of A549 cells in time- and concentration-dependent manners. The proportion of apoptosis, G0/G1 and G2/M phase increased in accordance with raising of the TSA concentration. The expression of p21 protein was significantly up-regulated and the expression of phosphorylation ERK was significantly down-regulated after A549 cells were treated with TSA.</p><p><b>CONCLUSIONS</b>Histone deacetylase inhibitor TSA can inhibit the proliferation of human lung cancer cell strains A549 and induce the cell cycle arrest and apoptosis in the A549 cells. This may be related to up-regulation of p21 protein expression and the down-regulation of phosphorylation ERK.</p>


Asunto(s)
Humanos , Adenocarcinoma , Metabolismo , Patología , Apoptosis , Ciclo Celular , Línea Celular Tumoral , Quinasas MAP Reguladas por Señal Extracelular , Metabolismo , Inhibidores de Histona Desacetilasas , Farmacología , Ácidos Hidroxámicos , Farmacología , Neoplasias Pulmonares , Metabolismo , Patología
13.
Chinese Journal of Oncology ; (12): 250-253, 2009.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-293139

RESUMEN

<p><b>OBJECTIVE</b>To study the effects of HDACi on the expression of survivin and NF-kappaB in human myeloma cell line U266 cells.</p><p><b>METHODS</b>U266 cells were cultured in RPMI 1640 in the presence of MS-275, and the cell viability was evaluated by trypan blue exclusion assay and cell count. The cell morphological changes were observed with Wright-Giemsa staining. The cell cycle was analyzed by flow cytometry, and the proteins of poly (ADP-ribose) polymerase (PARP), caspase-3, survivin, p21, CDK4 and IkappaB-alpha were detected by Western blotting.</p><p><b>RESULTS</b>MS-275 inhibited the growth of U266 cells in a dose- and time-dependent manner. The cell cycle was arrested at G(0)/G(1) phase. After exposure at 1.39 micromol/L MS-275 for 48 hours, the cell viability was decreased to 50%. The cell ratios of G(0)/G(1) phase were increased to (64.57 +/- 4.09)% for 24 h and (87.20 +/- 2.83)% for 36 h after 2 micromol/L MS-275 treatment. The visible morphological changes of U266 cells were confirmed with Wright-Giemsa staining. Cleaved-PARP, increased expression of p21, downregulation of expression of survivin, CDK4 and the phosphorylation of IkappaB-alpha was found by Western blot in MS-275 treated U266 cells.</p><p><b>CONCLUSION</b>MS-275-induced apoptosis of U266 cells is mediated by downregulation of expression of survivin and inactivation of NF-kappaB survival signalling pathways.</p>


Asunto(s)
Humanos , Apoptosis , Benzamidas , Farmacología , Caspasa 3 , Metabolismo , Ciclo Celular , Línea Celular Tumoral , Supervivencia Celular , Quinasa 4 Dependiente de la Ciclina , Metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Metabolismo , Relación Dosis-Respuesta a Droga , Inhibidores de Histona Desacetilasas , Farmacología , Proteínas Inhibidoras de la Apoptosis , Proteínas Asociadas a Microtúbulos , Metabolismo , Mieloma Múltiple , Metabolismo , Patología , FN-kappa B , Metabolismo , Poli(ADP-Ribosa) Polimerasas , Metabolismo , Piridinas , Farmacología , Transducción de Señal
14.
Chinese Journal of Pediatrics ; (12): 185-189, 2008.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-326188

RESUMEN

<p><b>OBJECTIVE</b>To survey the sleep habits (bedtime, wake time), sleep duration, and sleep problems in school-age children of China.</p><p><b>METHOD</b>From November to December, 2005, a total of 19,299 school-age children from 55 elementary schools of 9 cities entered the study by a cross-sectional survey. A parent-administered questionnaire and the Chinese version of the Children's Sleep Habits Questionnaire were applied to investigate children's sociodemographic characteristics and sleep behaviors, respectively.</p><p><b>RESULTS</b>The mean sleep duration was 9 hours and 10 minutes (9:10, SD:48 min) during the weekdays and 9:48 (SD: 63 min) during the weekends. In about 71.4% and 41.8% school-aged children the sleep duration per day did not reach the lowest criterion of 10 hours recommended by the Ministry of Education of China during weekdays and weekends, respectively. Sleep problems were common with prevalence ranging from 14.5% for sleep-disordered breathing to 75.3% for daytime sleepiness. Parasomnia (chi(2) = 13.76, P < 0.01) and sleep-disordered breathing (chi(2) = 119.83, P < 0.01) were more prevalent in boys than in girls; however, sleep anxiety was more prevalent in girls than in boys (chi(2) = 19.42, P < 0.01). Except for night waking, other types of sleep problems were significantly associated with age.</p><p><b>CONCLUSIONS</b>Inadequate sleep duration and sleep problems prevail among school-age children, which indicates that children's sleep health may be a major public health concern in China.</p>


Asunto(s)
Niño , Femenino , Humanos , Masculino , China , Epidemiología , Estudios Transversales , Hábitos , Sueño , Fases del Sueño , Trastornos del Sueño-Vigilia , Epidemiología , Estudiantes , Encuestas y Cuestionarios , Población Urbana
15.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-267893

RESUMEN

This study was aimed to investigate the possible influence of a novel E3 ubiquitin ligase CHIP (carboxyl terminus of Hsc70/Hsp70-interacting protein) on biological characteristics of cancer cells. Stable overexpression models in CML K562 cells were established via lipofectamine-mediated wild type CHIP and its TPR or U-box deletion mutants gene transfection. Followed G418 pressure selection, K562-CHIP stable transfected cell clones were obtained by limited dilution. The proliferation status and cell cycle were observed by MTT assay and FACS. The expression of related proteins and morphological changes were detected by Western blot and Wright-Giemsa staining. The results showed that overexpression of wild type CHIP did not inhibit cell proliferation, but slightly increased cell ratio of G(2)/M phase. CHIP gene had no effect on the stability of BCR-ABL kinase protein. HDAC inhibitor FK228-induced BCR-ABL degradation did not enhanced by CHIP. Notably the enlarged cells and abnormal mitotic cells remarkably increased in K562 WT-CHIP cells, indicating that CHIP may involve in the regulation of mitotic process. It is concluded that wild type CHIP induces mitotic abnormity in K562 cells.


Asunto(s)
Humanos , Proteínas de Choque Térmico , Genética , Metabolismo , Células K562 , Mitosis , Proteínas de Complejo Poro Nuclear , Genética , Metabolismo , Proteínas Proto-Oncogénicas , Genética , Metabolismo , Eliminación de Secuencia , Transfección , Ubiquitina-Proteína Ligasas , Genética , Metabolismo
16.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-230286

RESUMEN

The study was aimed to investigate the molecular mechanisms of histone deacetylase inhibitor SAHA-induced apoptosis of acute myeloid leukemia cell line HL-60. The effect of SAHA on HL-60 cell proliferation was detected by MTT assay and the cell morphological changes were observed with Wright-Giemsa and Hoechst33342 staining. The cell cycle distribution was determined by flow cytometry and the expression of cell signaling proteins were detected by Western-blot analysis. The results showed that SAHA inhibited the proliferation of HL-60 cells in dose- and time-dependent manners, after 2 micromol/L SAHA exposure for 12 - 48 hours, the cell cycle was arrested at G(0)/G(1) phase and apoptotic cell death was confirmed by either defined apoptotic bodies stained by Hoechst33342, Western blot showed cleaved-PARP, which represents the activation of caspase 3. The Western blot analysis indicated the activation of two important survival signal pathways after SAHA treatment, the phosphorylation of Raf and its downstream ERK kinases were remarkable downregulated, whereas the phosphorylation of AKT and its downstream molecular mTOR were not changed. It is concluded that SAHA-induced apoptosis of HL-60 cells is mediated by inactivation of p44/42 MAPK signaling pathway.


Asunto(s)
Humanos , Apoptosis , Células HL-60 , Inhibidores de Histona Desacetilasas , Ácidos Hidroxámicos , Farmacología , Sistema de Señalización de MAP Quinasas , Proteína Quinasa 1 Activada por Mitógenos , Metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Metabolismo , Proteínas Quinasas Activadas por Mitógenos , Metabolismo , Transducción de Señal
17.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-230274

RESUMEN

To investigate the possible involvement of Ras signaling in the hematopoietic differentiation of embryonic stem cells (ES cells), ES cells were transfected with RasN17, the dominant-negative mutant of Ras. Western blot was used to test the effect of RasN17 expression on Erk1/2 and Akt phosphorylation, semi-quantitative RT-PCR was used to detect expression of gene related to hematopoiesis in differentiation of ES cells. The results showed that the expression of RasN17 in the ES cells remarkably downregulated the phosphorylation of Erk1/2 and Akt simultaneously. Moreover, the expression of several markers related with hematopoiesis including Runx1, SCL and beta-major globin, were significantly suppressed in the EB expressing RasN17, whereas the transcription of Flk1, a gene required earlier than SCL in development of hematopoietic and endothelial lineages, was not influenced. It is concluded that the activation of Ras is pivotal for in vitro hematopoietic differentiation of ES cells.


Asunto(s)
Animales , Ratones , Diferenciación Celular , Fisiología , Células Cultivadas , Células Madre Embrionarias , Biología Celular , Genes ras , Hematopoyesis , Fisiología , Células Madre Hematopoyéticas , Biología Celular , Proteína Quinasa 3 Activada por Mitógenos , Metabolismo , Fosforilación , Transducción de Señal , Proteínas ras , Fisiología
18.
Artículo en Inglés | WPRIM (Pacífico Occidental) | ID: wpr-317017

RESUMEN

In the post-genomic era, identification of specific regulatory motifs or transcription factor binding sites (TFBSs) in non-coding DNA sequences, which is essential to elucidate transcriptional regulatory networks, has emerged as an obstacle that frustrates many researchers. Consequently, numerous motif discovery tools and correlated databases have been applied to solving this problem. However, these existing methods, based on different computational algorithms, show diverse motif prediction efficiency in non-coding DNA sequences. Therefore, understanding the similarities and differences of computational algorithms and enriching the motif discovery literatures are important for users to choose the most appropriate one among the online available tools. Moreover, there still lacks credible criterion to assess motif discovery tools and instructions for researchers to choose the best according to their own projects. Thus integration of the related resources might be a good approach to improve accuracy of the application. Recent studies integrate regulatory motif discovery tools with experimental methods to offer a complementary approach for researchers, and also provide a much-needed model for current researches on transcriptional regulatory networks. Here we present a comparative analysis of regulatory motif discovery tools for TFBSs.


Asunto(s)
Algoritmos , Secuencias de Aminoácidos , Secuencia de Bases , Sitios de Unión , Biología Computacional , Internet , Datos de Secuencia Molecular , Unión Proteica , Elementos Reguladores de la Transcripción , Genética , Análisis de Secuencia de ADN , Métodos , Homología de Secuencia de Ácido Nucleico , Factores de Transcripción , Metabolismo
19.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-270507

RESUMEN

<p><b>OBJECTIVE</b>To study the effects on the expression of mGluR5 mRNA and protein levels in primarily cultured hippocampal neurons after lead exposure.</p><p><b>METHODS</b>Primary embryonic rat hippocampal neuronal culture was prepared. On the 3(rd) day of incubation, lead chloride solution was added into medium to produce four different lead exposure levels: 0, 1 x 10(-8) mol/L, 1 x 10(-6) mol/L, 1 x 10(-4) mol/L Pb(2+). After 10 days of incubation, the neurons were collected to measure the alteration of mGluR5 mRNA expression by real-time fluorescent quantity PCR and the expression of mGluR5 in protein level by Western blot.</p><p><b>RESULTS</b>The studies revealed that mGluR5 mRNA expression was down-regulated after lead exposure in a dose-dependent manner. The mGluR5 mRNA expression of the lower lead-exposed neurons (Pb(2+) 10(-8) mol/L), the medium lead-exposed neurons(Pb(2+) 10(-6) mol/L), the higher lead-exposed neurons(Pb(2+) 10(-4) mol/L) were 0.724, 0.421, 0.321 times less than that of the controls, respectively. The Western blot demonstrated that mGluR5 expression in protein level should be decreased after lead exposure.</p><p><b>CONCLUSION</b>The expression of mGluR5 in mRNA and protein levels should be down-regulated after lead exposure at different lead levels in a dose-dependent manner.</p>


Asunto(s)
Animales , Femenino , Embarazo , Ratas , Animales Recién Nacidos , Células Cultivadas , Hipocampo , Metabolismo , Plomo , Toxicidad , Intoxicación del Sistema Nervioso por Plomo , Metabolismo , Neuronas , Metabolismo , ARN Mensajero , Genética , Ratas Sprague-Dawley , Receptor del Glutamato Metabotropico 5 , Receptores de Glutamato Metabotrópico , Genética
20.
Chinese Journal of Pediatrics ; (12): 176-180, 2007.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-356206

RESUMEN

<p><b>OBJECTIVE</b>To develop the Chinese version of Children's Sleep Habits Questionnaire (CSHQ) by translating and retranslating the English version of CSHQ and to examine the reliability, content validity, construct validity of the Chinese version in a general population of school-age children.</p><p><b>METHODS</b>During November and December 2005, a total of 20 457 school-age children from 55 primary schools of 9 cities were recruited through cluster-stratified selection, using geographic location, economic standard, and population density as criteria. A parent-administered questionnaire and the Chinese version of CSHQ were completed to quantify sociodemographic characteristics and to characterize sleep patterns and sleep disturbances, respectively.</p><p><b>RESULTS</b>The internal consistency of overall questionnaire and the eight subscales of the Chinese version of CSHQ was good (Cronbach's alpha coefficient was 0.73 and ranged from 0.42 to 0.69, respectively). The consistency between mother and father was ICCs = 0.89 for overall questionnaire and ranged from ICCs = 0.83 to ICCs = 0.92 for subscales. The test-retest reliability was ICCs = 0.85 for overall questionnaire and range from ICCs = 0.60 to ICCs = 0.88 for subscales. Factor analysis revealed a three-factor structure, which could represent bedtime behavior problems, sleep disturbance, sleep duration and daytime sleepiness, respectively, and the three factors could be used to explain 58.63% of the total variance. The factor loading was above 0.5 for the corresponding subscale.</p><p><b>CONCLUSIONS</b>Reliability, content validity and construct validity of the Chinese version of CSHQ in a general population of school-age children are good. It appears to be a suitable instrument for measuring sleep patterns and screening for sleep problems in school-aged children.</p>


Asunto(s)
Niño , Femenino , Humanos , Masculino , China , Epidemiología , Psicometría , Reproducibilidad de los Resultados , Sueño , Trastornos del Sueño-Vigilia , Epidemiología , Encuestas y Cuestionarios
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