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In this work, a sensitive and efficient method was established and validated for qualitative and quantitative analysis of major quassinoid diterpenoids constituents from the extract of Eurycoma longifolia by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and ultra-performance liquid chromatography coupled with a triple quadrupole mass spectrometry(UPLC-QQQ-MS/MS). The UPLC-Q-TOF-MS analysis was performed on an Agilent Eclipse Plus C_(18) RRHD(2.1 mm×100 mm,1.8 µm) column with acetonitrile and 0.1% formic acid water as mobile phase by gradient elution. The UPLC-QQQ-MS/MS analysis was performed on an Agilent Eclipse Plus C_(18) RRHD(2.1 mm×50 mm, 1.8 µm)column with acetonitrile and 0.1% formic acid water as mobile phase by gradient elution. The data was collected by electrospray ionization in positive mode. According to the contrast of the reference standards and the accurate masses of molecules, a total of 32 quassinoid diterpenoids in E. longifolia extract were identified by UPLC-Q-TOF-MS. For quantitative the linear range of 4 detected quassinoid diterpenoids were good(r≥0.999 6), and the overall recoveries ranged from 90.35% to 106.4%, with the RSD ranging from 1.8% to 3.6%. The method was accurate, reliable and efficient, and could comprehensively reflect the chemical constituents and content of E. longifolia, and could provide a reference for further elucidating its pharmacological basis and quality control.
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Medicamentos Herbarios Chinos , Eurycoma , Cuassinas , Cromatografía Líquida de Alta Presión , Diterpenos , Espectrometría de Masas en TándemRESUMEN
Through routine and nested PCR amplifications, four complete genome sequences of porcine Torque teno virus (TTV) type II were obtained from swine herds. By comparison with the TTV genome sequences deposited in GenBank, we found the most divergent types so far described. The level of genetic diversity between these genomes is higher than would be expected within a single virus species. A nucleotide and amino acid phylogenetic tree was constructed.
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Genoma Viral , Torque teno virus/genética , Animales , ADN Viral , Bases de Datos Genéticas , Genes Virales , Variación Genética , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Filogenia , PorcinosRESUMEN
Here, we report three complete genome sequences of porcine torque teno virus type I (TTV1) which were obtained from swine tissues and sera from southern China through routine and nested PCR amplification and characterized together with other genome sequences already deposited in GenBank. The results showed that the TTV1 sequences were highly divergent and could be divided into 1a and 1b subtypes.
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ADN Viral/química , ADN Viral/genética , Genoma Viral , Torque teno virus/genética , Animales , China , Genotipo , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Porcinos , Torque teno virus/clasificación , Torque teno virus/aislamiento & purificaciónRESUMEN
OBJECTIVE: To study the inhibiting effect of Endostar combined with ginsenoside Rg3 on breast cancer tumor growth in tumor-bearing mice. METHODS: Female mice were selected as experimental animals, and breast cancer tumor-bearing mouse models were established and then divided into groups A, B, C and D that respectively received saline, recombinant human endostatin, ginsenosides Rg3 and recombinant human endostatin combined with Rg3 intervention; 7 d, 14 d and 21 d after intervention, tumor tissue volume was measured; 21 d after intervention, mice were killed, tumor tissue was collected, and mRNA contents of angiogenesis molecules, invasion molecules, autophagy marker molecules and autophagy signaling pathway molecules were detected. RESULTS: At 7 d, 14 d and 21 d after intervention, tumor tissue volume of groups B, C and D was lower than that of group A, and tumor tissue volume of group D was lower than that of groups B and C; mRNA contents of VEGFA, VEGFB, VEGFC, MMP2, MMP9, p62, mTOR, PI3K, Akt, JNK and Beclin-1 in tumor tissue of groups B, C and D were significantly lower than those of group A, and LC3-II/LC3-I was significantly higher than that of group A; mRNA contents of VEGFA, VEGFB, VEGFC, MMP2, MMP9, p62, mTOR, PI3K, Akt, JNK and Beclin-1 in tumor tissue of group D were significantly lower than those of groups B and C, and LC3-II/LC3-I was higher than that of groups B and C. CONCLUSIONS: Endostar combined with ginsenoside Rg3 has stronger inhibiting effect on breast cancer tumor growth in tumor-bearing mice than single drug, and it can inhibit angiogenesis and cell invasion, and enhance cell autophagy.
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Recently pseudorabies outbreaks have occurred in many vaccinated farms in China. To identify genetic characteristics of pseudorabies virus (PRV) strains, we obtained the genomic sequences of PRV strains HeN1 and JS, which were compared to 4 PRV genomes and 729 partial gene sequences. PRV strains isolated in China showed marked sequence divergence compared to European and American strains. Phylogenetic analysis revealed that for the first time PRV can be divided into 2 distinct clusters, with Chinese strains being genotype II and PRVs isolated from other countries being genotype I. Restriction fragment length polymorphism analysis confirmed differences between HeN1 and Bartha strains, as did the presence of unique insertion/deletion polymorphisms and microsatellites. This divergence between the two genotypes may have been generated from long-term, independent evolution, which could also explain the low efficacy of the Bartha vaccine in protecting pigs infected with genotype II PRV.
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Variación Genética , Herpesvirus Suido 1/clasificación , Herpesvirus Suido 1/genética , Seudorrabia/epidemiología , Seudorrabia/virología , Animales , China/epidemiología , Análisis por Conglomerados , ADN Viral/química , ADN Viral/genética , Genoma Viral , Genotipo , Herpesvirus Suido 1/aislamiento & purificación , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia , PorcinosRESUMEN
One sediment core was obtained from the center of Yamenqi Lake at the Songnen Plain in eastern China in August 2010 using a gravity corer. The sediment samples were digested using HNO3-HClO4-HF. Concentrations of the metals Cr, Cu, Mn, Ni, Fe, Pb, Zn, Cd, Al, Ba, Ca, K, Li, Mg, Na and Sr were determined using inductively coupled plasma-mass spectrometry (ICP-MS). With the dating approach, the characteristics and polluted history of heavy metals in lake sediments were determined. The heavy metal pollution of sediment was discussed based on the enrichment factor. The results are listed as following:(1) the contents of all elements had a smooth variation trend before 1950,and fluctuated severely from 1950 to 1990. The contents of Mn, Zn, Cd, Pb, Fe, Ni, Cr, Cu, Ca, Li and Sr increased obviously since 1990. (2) Mn, Zn, Pb and Cd showed light contamination degree. Contents of Ni, Cr and Cu were below the contamination level.