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Li P.,Nanjing Agricultural University | Han H.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Zhai X.,Jiangsu Animal Husbandry and Veterinary College | He W.,Jiangsu Animal Husbandry and Veterinary College | And 2 more authors.
Journal of Chromatographic Science | Year: 2012

An isocratic reversed-phase high-performance liquid chromatography method with UV detection is developed and validated for the simultaneous determination of ketamine, xylazine, and midazolam in canine plasma. Analytes are extracted from alkalinized samples into diethyl ethermethylene chloride (7:3, v:v) using single-step liquidliquid extraction. Chromatographic separation is performed on a C18 column using a mobile phase containing an acetonitrilemethanol- 10 mM sodium heptanesulfonate buffer adjusted to pH 3, with glacial acetic acid (44:10:46, v:v) at a detection wavelength of 210 nm, with a total runtime of 10 min. The calibration is linear over the range of 78.125-5000 ng/mL for ketamine and 15.625-1000 ng/mL for xylazine and midazolam. The limits of detection are 17.8, 10.3, and 15.1 ng/mL for ketamine, xylazine, and midazolam, respectively. The extraction recoveries are 76.1 for ketamine, 91.0 for midazolam, and 78.2 for xylazine. The method is successfully used for clinical and pharmacokinetic studies of the three-drug fixed dose combination formulations. © 2012 The Author. Source


Xiong A.S.,State Key Laboratory of Crop Genetics and Germplasm Enhancement
BMC genomics | Year: 2014

MicroRNAs (miRNAs) are small, non-coding RNAs of 20 to 24 nucleotides that regulate gene expression and responses to biotic and abiotic stress. Till now, no reports have previously been published concerning miRNAs in celery. Two small RNAs libraries were constructed from two celery varieties, 'Jinnan Shiqin' and 'Ventura', and characterized by deep sequencing. A total of 431 (418 known and 13 novel) and 346 (341 known and five novel) miRNAs were identified in celery varieties 'Jinnan Shiqin' and 'Ventura', respectively. Potential miRNA-target genes were predicted and annotated by screening diverse protein databases, including Gene Ontology, Cluster of Orthologous Groups and Kyoto Encyclopedia of Genes and Genomes. Significant differential expression between the two varieties was seen for 221 miRNAs. qRT-PCR was used to analyze the abundance of six miRNAs under cold and heat stress conditions. The results showed that miRNAs may have important functions in controlling temperature stress in celery. A large number of miRNAs were identified in celery, and their target genes, functional annotations, and gene expression patterns have been explored.These findings provide the first information on celery miRNAs and enhance understanding of celery miRNA regulatory mechanisms under extreme temperature stress. Source


Shi L.,Nanjing Agricultural University | Qin L.,Nanjing Agricultural University | Xu Y.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Ren A.,Nanjing Agricultural University | And 4 more authors.
Molecular Biology Reports | Year: 2012

This study investigated the role of the mevalonate pyrophosphate decarboxylase gene in the triterpene biosynthetic pathway of Ganoderma lucidum. The mevalonate pyrophosphate decarboxylase gene (mvd) was isolated using a degenerate primer-PCR technique. An analysis of the Gl-mvd transcription profile revealed a positive correlation between the expression of the Gl-mvd gene and triterpene content changes in G. lucidum during development. Furthermore, a promoter deletion analysis was conducted in G. lucidum to investigate the promoter activity and the role of methyl jasmonate (MeJA) responsive elements in the mvd promoter under the MeJA elicitor. The overexpression of Gl-mvd increased triterpene accumulation compared with the wild-type strain and increased the expression of several genes involved in the triterpene biosynthetic pathway. The findings of this study suggest that mvd may play an important role in triterpene biosynthesis regulation. Moreover, there may be the interactions among the genes involved in the triterpene biosynthetic pathway in the G. lucidum. Additionally, this study provides an approach for improving triterpene content through the overexpression of a key gene. © Springer Science+Business Media B.V. 2011. Source


Sun C.-Q.,Nanjing Agricultural University | Chen F.-D.,Nanjing Agricultural University | Teng N.-J.,Nanjing Agricultural University | Teng N.-J.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | And 4 more authors.
Euphytica | Year: 2010

Chrysanthemum grandiflorum 'Yuhuaxingchen' is an important commercial chrysanthemum cultivar with excellent ornamental quality but low drought tolerance, whereas C. indicum has exceptional drought tolerance. In our earlier study, many hybrid seeds between them were obtained through interspecific hybridization. In the present study, we selected six putative hybrid lines with most drought tolerance from all the hybrid lines by withholding water, indentified their facticity by chromosome counting, and then evaluated their drought tolerance through determining foliar electrolyte leakage (EL), contents of malondialdehyde (MDA) and proline, and plant survival rate after 20% polyethylene glycol 6000 treatment. It was found that 155 out of 282 seeds germinated and only 132 seedlings survived. In addition, chromosome and morphological analysis showed that the six putative hybrids were real hybrids and their morphological features were intermediate between their parents. Furthermore, the density of leaf epidermal hair, proline content, and plant survival rate were the highest in C. indicum and the lowest in C. grandiflorum among the six hybrids and their parents. In contrast, EL value and MDA content were the highest in C. grandiflorum and the lowest in C. indicum. These results suggest that some true hybrids with improved drought tolerance can be obtained through interspecific hybridization in chrysanthemum breeding. Therefore, interspecific hybridization between chrysanthemum cultivars and their wild species may become a promising way to improve their biotic and abiotic resistance in the future breeding. © 2010 Springer Science+Business Media B.V. Source


Wang X.H.,Nanjing Agricultural University | Wang X.H.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Song Y.P.,Nanjing Agricultural University | Song Y.P.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | And 10 more authors.
Journal of Animal and Plant Sciences | Year: 2013

Non-heading Chinese cabbage is one of the main vegetables in Asia, especially in China. To analyze the tolerance of non-heading Chinese cabbage to copper (Cu) stress, 10μM, 100μM and 1mM copper were used to treat two cultivars of 'Wutacai' as well as 'Erqing' and then photosynthesis, chlorophyll fluorescence parameters, biomass and chlorophyll content were investigated. In our study, 1mM copper significantly inhibited the photosynthesis and plant growth of both two cultivars. While at 100μM copper, the growth inhibition was just found in 'Erqing', not in 'Wutacai'. The inhibition of copper on the photosynthesis was not only by stomatal factors but also by non-stomatal factors. Copper stress caused damage of photosystem resulting in reduction of PSII efficiency while non-photochemical-quenching parameter NPQ increased. Besides, at low concentration with 10μM, copper promoted the photosynthesis and plant growth. Therefore, the copper tolerance of non-heading Chinese cabbage was dependent on the genotype, and the plant growth inhibition under copper stress was due to the damage of photosystem. Our results will provide a theoretical basis for a further study on non-heading Chinese cabbage tolerance to copper and will be helpful for the production of non-heading Chinese cabbage in heavy metals contaminated areas. Source

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