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1.
Bing Du Xue Bao ; 30(4): 382-6, 2014 Jul.
Artículo en Chino | MEDLINE | ID: mdl-25272590

RESUMEN

Rolling circle amplification (RCA) is a newly developed experimental technique that can specific ally amplify circular DNA. Since 2008, RCA has been extensively used in hepatitis B virus (HBV) research, such as the amplification of the full-length sequence of the HBV genome, and the analysis of the drug-resistant mutations of HBV covalently closed circular DNA (cccDNA), amongst others. To create an easy assay for the analysis of duck hepatitis B virus (DHBV) cccDNA, this study established an RCA-based method. DHBV cccDNA was amplified from the DHBV DNA samples of duck liver with four pairs of sulfur-modified primers, which were designed according to the highly conserved sequence of DHBV using sera DHBV DNA as the negative control. DHBV cccDNA was detected in the obtained RCA products by the sequencing of RCA amplicons that were amplified with primer pairs on both sides of the gap of DH BV relaxed circular DNA, rather than by digesting RCA products with a restriction enzyme. The liver and sera DHBV DNA samples of 39 ducks infected with DHBV were examined with the RCA-based DHBV cccDNA detection method, and the results showed that while DHBV cccDNA was detected from all 39 liver DHBV DNA samples, no DHBV cccDNA was found in any of the sera DHBV DNA samples. These results suggest that the method established in the study is highly specific and sensitive for the detection of DHBV cccDNA. The establishment of this RCA-based DHBV method for cccDNA detection lays the groundwork for using a DHBV model to study the role of cccDNA in the pathogenesis of hepatitis B and to evaluate the effect of anti-virus therapies.


Asunto(s)
ADN Circular/genética , ADN Viral/genética , Infecciones por Hepadnaviridae/veterinaria , Virus de la Hepatitis B del Pato/aislamiento & purificación , Enfermedades de las Aves de Corral/virología , Animales , Cartilla de ADN/genética , Patos , Infecciones por Hepadnaviridae/virología , Virus de la Hepatitis B del Pato/genética , Hígado/virología , Reacción en Cadena de la Polimerasa/métodos
2.
Huan Jing Ke Xue ; 32(10): 3106-12, 2011 Oct.
Artículo en Chino | MEDLINE | ID: mdl-22279931

RESUMEN

The chemical-enhanced washing of Cu2+ or/and phenanthrene (PHE) single or combined contaminated loess soil in Gansu Province was investigated with disodium ethylene diamine tetraacetate (EDTA) or/and sodium dodecyl sulfate (SDS) by the batch equilibrium experiments. The experimental results showed that EDTA or/and SDS could remove efficiently Cu2+ and/or PHE in single-contaminated or combined contaminated loess soils. The Cu2+ removal was significantly promoted by coexisting PHE with low concentration of EDTA (EDTA < 0.1 mol/L), however, the removal was slightly hindered with high concentration of EDTA (EDTA > 0.1 mol/L). As for the PHE removal by EDTA, it was founded that coexisting Cu2+ could enhance the PHE removal in the investigated ranges of the concentrations of EDTA. When concentration of EDTA was 0.01 mol/L, the removal of combined PHE was 20.94% higher than that of single PHE. The experimental results of the removal of contaminations by SDS showed that coexisting Cu2+ could suppress slightly the removal of PHE at a concentration of less than 4 000 mg/L SDS, but could assist the removal of PHE in 5 000 mg/L or higher SDS concentration. On the contrary, the influence of coexisting PHE for the removal of Cu2+ by SDS was that it facilitated Cu2+ extraction by low concentrations of SDS, however, it inhibited the removal of Cu2+ at high concentrations of SDS. The removal efficiencies of PHE and Cu2+ were improved greatly as using combined EDTA-SDS. Beside, there are some differences in the removal efficiency of the oth contaminants with the different sequence of EDTA and SDS added in the washing solution. In EDTA washing followed by SDS, SDS washing followed by EDTA and mixture of SDS-EDTA washing concurrently, the removal of Cu2+ is 91.40%, 95.10% and 96.50%, respectively, which is 28.46%, 32.16%, 33.56% higher than that of combined, 62.94%, by single EDTA. For PHE, the removal is 68.30%, 85.40%, 84.95%, respectively, which is 16.14%, 33.24%, 32. 79% higher than that of combined PHE, 52.16%, by single SDS. Thus, SDS washing followed by EDTA or mixture of SDS-EDTA washing concurrently is considered as the optimal washing sequence for PHE and Cu2+ removal.


Asunto(s)
Cobre/aislamiento & purificación , Restauración y Remediación Ambiental/métodos , Fenantrenos/aislamiento & purificación , Contaminantes del Suelo/aislamiento & purificación , China , Cobre/química , Ácido Edético/química , Fenantrenos/química , Dodecil Sulfato de Sodio/química , Contaminantes del Suelo/química
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