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Non-small cell lung cancer (NSCLC) is still an unresolved source of tumor-related death internationally. Current studies have discovered that microRNAs (miRNAs) are associated with diverse cancers development, including NSCLC. Our paper focused on the functional character of miR-4286 in NSCLC. miR-4286 level in 68 cases of NSCLC tissues, matched neighboring nontumor tissues and different cancer cell lines were inspected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The connection concerning miR-4286 expression and clinicopathological features of patients with NSCLC were further determined. After knockdown or overexpression of miR-4286, cell viability, cell cycle, and/or apoptotic cells were examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assay, respectively. Moreover, the cell cycle- and apoptosis-related proteins were estimated by qRT-PCR and Western blot. In comparison with the matched nontumor tissues, miR-4286 was significantly enhanced in lung malignancy tissues and different cell lines. miR-4286 expression was related with the tumor-node-metastasis stage, lymphatic metastasis, and distant metastasis. Cell viability was ominously weakened by suppression of miR-4286 in A549 cells, whereas was statistically upregulated by overexpression of miR-4286 in NCI-H1299 cells. Additionally, we detected that suppression of miR-4286 tempted cell cycle arrest in G1 stage and fortified apoptosis in A549 cells. Runx3 was recognized as one target gene of miR-4286, and the impacts of suppression of miR-4286 on cell viability and apoptosis were through regulation of Runt-related transcription factor 3. Our study suggests that miR-4286 overexpression represents a tumor promoter role in NSCLC cells.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/genética , Subunidad alfa 3 del Factor de Unión al Sitio Principal/genética , MicroARNs/genética , Pronóstico , Células A549 , Adulto , Anciano , Apoptosis/genética , Biomarcadores de Tumor/genética , Carcinoma de Pulmón de Células no Pequeñas/epidemiología , Carcinoma de Pulmón de Células no Pequeñas/patología , Puntos de Control del Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Supervivencia sin Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Invasividad Neoplásica/genética , Invasividad Neoplásica/patologíaRESUMEN
OBJECTIVE: To screen and characterize the aptamer of Escherichia coli outer member protein tolC. METHODS: By using the recombinant E.coli outer member protein tolC for the screening target, oligonucleotides which were capable of specifically binding to the protein were screened from a random oligonucleotide library through the stematic evolution of ligand by exponential enrichment (SELEX) technique. The binding capacity of ssDNA to the targeted protein from each round was detected by the FITC fluorescence labeling technique.The ssDNA from the last cycle was cloned and sequenced,and the second structure was further analyzed by the DNAMan program. RESULTS: After 12 cycles of selection, 40 clones were selected randomly and sequenced. Although a unique conserved sequence was not obtained among the 23 obtained aptamers by the primary structure analysis,three pairs of aptamers and two pairs of aptamers were found to be identical.Analysis of the secondary structure revealed that the stem-loop and bulge loop were the main motifs,indicating that they might play a key role in the binding of aptamers to the target protein. According to the characteristic of the second structure,23 aptamers were divided into four families,and aptamer 20 bore the greatest affinity. CONCLUSION: Aptamers against E.coli outer member protein tolC were successfully identified by the SELEX method. The results laid a foundation for the investigation of the interference to the drug resistance of E. coli and the underlying mechanisms.
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Aptámeros de Nucleótidos/genética , Proteínas de la Membrana Bacteriana Externa/química , Proteínas de Escherichia coli/química , Escherichia coli , Proteínas de Transporte de Membrana/química , ADN de Cadena Simple , Biblioteca de Genes , Proteínas Recombinantes/química , Técnica SELEX de Producción de AptámerosRESUMEN
The eddy covariance (EC) technique has emerged as the most widely used method for long-term continuous methane flux (FCH4) observations. However, the completeness of the FCH4 time series is limited by instrumental failures and data quality issues, resulting in missing data gaps ranging from 20 % to 90 %. In this situation, the excellent performance of machine learning (ML) algorithms in filling missing FCH4 data has provided a foundation for developing regional-scale FCH4 models. In this study, we established estimation models for FCH4 utilizing random forest (RF), support vector machine (SVM), back propagation (BP) and nonlinear multiple regression (MLR) algorithms. The maximal information coefficient (MIC) technique was employed to identify and rank the environmental factors that were correlated with FCH4. Our findings revealed that soil temperature (Ts), soil water content (SWC) and air temperature (Ta) were the primary environmental factors influencing FCH4. Among the four algorithms, from perspectives of model accuracy and relatively small number of driving factors, the RF models exhibited the best performance, followed by BP and SVM, whereas MLR demonstrated the lowest performance. Among the 144 RF models established using nine datasets, RF model with 8 driving factors in all-year (RFall-year8) could capture seasonal variations. Ultimately, we recommend (RFall-year8 as the optimal model for estimating FCH4 in the Dajiuhu subalpine peatland.
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Peatlands play a crucial role in mediating the emissions of methane through active biogeochemical cycling of accumulated carbon driven by methane-metabolizing microorganisms; meanwhile, they serve as vital archives of atmospheric heavy metal deposition. Despite many edaphic factors confirmed as determinants to modulate the structure of methanotrophic and methanogenic communities, recognition of interactions among them is limited. In this study, peat soils were collected from Dajiuhu peatland to assess the presence of heavy metals, and methanotrophs and methanogens were investigated via high-throughput sequencing for functional genes mcrA and pmoA. Further analyses of the correlations between methane-related functional groups were conducted. The results demonstrated that both methane-metabolizing microorganisms and heavy metals have prominent vertical heterogeneity upward and downward along the depth of 20 cm. Pb, Cd, and Hg strongly correlated with methanotrophs and methanogens across all seasons and depths, serving as forceful factors in structural variations of methanogenic and methanotrophic communities. Particularly, Pb, Cd, and Hg were identified as excessive elements in Dajiuhu peatland. Furthermore, seasonal variations of networks among methane-related functional groups and environmental factors significantly affected the changes of methane fluxes across different seasons. Concretely, the complicated interactions were detrimental to methane emissions in the Dajiuhu peatland, leading to the minimum methane emissions in winter. Our study identified the key heavy metals affecting the composition of methane-metabolizing microorganisms and linkages between seasonal variations of methane emissions and interaction among heavy metals and methane-metabolizing microorganisms, which provided much new reference and theoretical basis for integrated management of natural peatlands.
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Mercurio , Metales Pesados , Metano , Cadmio , Plomo , Suelo/químicaRESUMEN
The rapid development of modern society has resulted in discharge of large, heavy metal quantities into wetlands that have been continuously accumulating, causing severe pollution. Dajiuhu, located in the Shennongjia Forest District of Hubei Province in China, is a wetland of significant value internationally, serving as a model wetland ecosystem with heightened scientific research value. In this study, 27 surface sediment samples from nine sub-lakes in Dajiuhu were collected in August 2020. The concentrations of Cd, Cr, Cu, Ni, Pb, and Zn in the sediments were determined. The heavy metal occurrence and speciation characteristics were analyzed by an improved BCR (European Community Bureau of Reference) extraction method. Four methods were used to evaluate heavy metals' pollution degree and ecological risk. The possible source of heavy metals was inferred using correlation analysis and principal component analysis. The heavy metal content in the lake sediments of Dajiuhu wetland was from the highest to the lowest concentration as follows: Zn [Formula: see text] Cr [Formula: see text] Ni [Formula: see text] Pb [Formula: see text] Cu [Formula: see text] Cd. The average Cd content exceeded the national nature reserve threshold values, while the other heavy metals measured were below their respective threshold values. However, due to the occurrence of Pb and Cd in different forms, they still pose certain pollution and ecological risk to the lake wetlands. On the other hand, Zn, Cr, Ni, and Cu do not pose an ecological risk in the lakes of the Dajiuhu wetland. The spatial distribution of heavy metal content in the nine sub-lakes did vary significantly. Regarding the heavy metal sources in the lake sediments, Ni, Cr, and Cu originate from natural factors, and Cd and Pb have mainly anthropogenic origins. In contrast, Zn has both natural and anthropogenic origins. This study provides further insights into the study of heavy metal pollution in lake wetlands. It provides a framework and a direction for managing heavy metal pollution in the Dajiuhu wetland.
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Metales Pesados , Contaminantes Químicos del Agua , Humedales , Ecosistema , Lagos/análisis , Cadmio/análisis , Plomo/análisis , Sedimentos Geológicos/análisis , Monitoreo del Ambiente/métodos , Contaminantes Químicos del Agua/análisis , Metales Pesados/análisis , Medición de Riesgo , ChinaRESUMEN
We collected evapotranspiration data of Dajiuhu peatland in Shennongjia from 2016 to 2017 with eddy covariance method and estimated the value of crop coefficient (Kc) using FAO56 Penman-Monteith equation and the linear relationship between actual evapotranspiration (ETa) and referenced evapotranspiration (ET0). We analyzed the characteristics of referenced evapotranspiration and its main influencing factors and calculated the crop coefficient of the wetland dominated by Sphagnum. The results showed that the daily averaged ETa were 1.63 and 1.38 mm·d-1 in 2016 and 2017, the daily averaged ET0 were 1.61 and 1.23 mm·d-1 in 2016 and 2017. Environmental factors influencing ET0 included net radiation, air temperature, vapor pressure deficit, wind speed, and relative humidity. The Kc values for the growing seasons of 2016, 2017, and 2016-2017 were 0.95 (R2 of linear regression between ETa and ET0 was 0.96), 1.03 (R2=0.95), and 0.98 (R2=0.95). The Kc values in 2016, 2017, and 2016-2017 were 0.92 (R2=0.94), 0.95 (R2=0.89), and 0.93 (R2=0.92). Kc was effective in the range of 0.92-1.03 for the wetland dominated by Sphagnum. The identified parameters could be widely used in studies on climate change, ecosystem services, and water management in peatlands.
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Ecosistema , Transpiración de Plantas , Productos Agrícolas , Temperatura , Agua , VientoRESUMEN
Biological invasion is regarded as one of the greatest environmental problems facilitated by globalization. Some hypotheses about the invasive mechanisms of alien invasive plants consider the plant-herbivore interaction and the role of plant defense in this interaction. For example, the "Shift Defense Hypothesis" (SDH) argues that introduced plants evolve higher levels of qualitative defense chemicals and decreased levels of quantitative defense, as they are released of the selective pressures from specialist herbivores but still face attack from generalists. Common groundsel (Senecio vulgaris), originating from Europe, is a cosmopolitan invasive plant in temperate regions. As in other Senecio species, S. vulgaris contains pyrrolizidine alkaloids (PAs) as characteristic qualitative defense compounds. In this study, S. vulgaris plants originating from native and invasive ranges (Europe and China, respectively) were grown under identical conditions and harvested upon flowering. PA composition and concentration in shoot and root samples were determined using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). We investigated the differences between native and invasive S. vulgaris populations with regard to quantitative and qualitative variation of PAs. We identified 20 PAs, among which senecionine, senecionine N-oxide, integerrimine N-oxide and seneciphylline N-oxide were dominant in the roots. In the shoots, in addition to the 4 PAs dominant in roots, retrorsine N-oxide, spartioidine N-oxide and 2 non-identified PAs were also prevalent. The roots possessed a lower PA diversity but a higher total PA concentration than the shoots. Most individual PAs as well as the total PA concentration were strongly positively correlated between the roots and shoots. Both native and invasive S. vulgaris populations shared the pattern described above. However, there was a slight trend indicating lower PA diversity and lower total PA concentration in invasive S. vulgaris populations than native populations, which is not consistent with the prediction of SDH.
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The main purpose of this study conducted from August 2010 was to find biomass and productivity of epilithic algae and their relations to environmental factors and try to explore the restrictive factors affecting the growth of algae in the Gufu River, the one of the branches of Xiangxi River located in the Three Gorges Reservoir of the Yangtze River, Hubei Province, Central China. An improved method of in situ primary productivity measurement was utilized to estimate the primary production of the epilithic algae. It was shown that in rivers, lakes, and reservoirs, algae are the main primary producers and have a central role in the ecosystem. Chlorophyll a concentration and ash-free dry mass (AFDM) were estimated for epilithic algae of the Gufu River basin in Three Gorges Reservoir area. Environmental factors in the Gufu River ecosystem highlighted differences in periphyton chlorophyll a ranging from 1.49 mg m-2 (origin) to 69.58 mg m-2 (terminal point). The minimum and maximum gross primary productivity of epilithic algae were 96.12 and 1439.89 mg C m-2 day-1, respectively. The mean net primary productivity was 290.24 mg C m-2 day-1. The mean autotrophic index (AFDM:chlorophyll a) was 407.40. The net primary productivity, community respiration ratio (P/R ratio) ranged from 0.98 to 9.25 with a mean of 2.76, showed that autotrophic productivity was dominant in the river. Relationship between physicochemical characteristics and biomass was discussed through cluster and stepwise regression analysis which indicated that altitude, total nitrogen (TN), NO3--N, and NH4+-N were significant environmental factors affecting the biomass of epilithic algae. However, a negative logarithmic relationship between altitude and the chlorophyll a of epilithic algae was high. The results also highlighted the importance of epilithic algae in maintaining the Gufu River basin ecosystems health.