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- 저자 : Yan Pi (검색결과 20 건)
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Molecular Cloning and Characterization of a Novel Stem-specific Gene from Camptotheca acuminata
Pi, Yan,Liao, Zhihua,Chai, Yourong,Zeng, Hainian,Wang, Peng,Gong, Yifu,Pang, Yongzhen,Sun, Xiaofen,Tang, Kexuan Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.1
In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.
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An optimized procedure for detection of genetically modified DNA in refined vegetable oils
Yuzhu Duan,Yan Pi,Changwen Li,Keji Jiang 한국식품과학회 2021 Food Science and Biotechnology Vol.30 No.1
In this study, the amplifiable DNA from refinedvegetable oils was isolated by using commercial DNAextraction kits based on the CTAB method in combinationwith nucleic acid enrichment, and then the presence ofgenetically modified (GM) soybean and maize DNA in theoils was traced by PCR. The results showed that theduration and intensity of heating had no significant effecton the DNA stability and concentration in oils for a shortperiod, suggesting that DNA in oils could be stablyreserved for a certain time, thus making it possible to tracedown refined vegetable oils reliably and effectively. Theresults provided a set of primers suitable for systematicGM oil detection. More importantly, this study made animportant contribution to the economical and reliabledetection of GM vegetable oils regarding food authenticityissues.
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Huang, Zhuoshi,Jiang, Keji,Pi, Yan,Hou, Rong,Liao, Zhihua,Cao, Ying,Han, Xu,Wang, Qian,Sun, Xiaofen,Tang, Kexuan Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.5
The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.
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Zheng Chao,Wu Yan,Liang Zhen Hua,Pi Jin Song,Cheng Shi Bin,Wei Wen Zhuo,Liu Jing Bo,Lu Li Zhi,Zhang Hao 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.2
Objective: Cage rearing has critical implications for the laying duck industry because it is convenient for feeding and management. However, caging stress is a type of chronic stress that induces maladaptation. Environmental stress responses have been extensively studied, but no detailed information is available about the comprehensive changes in plasma metabolites at different stages of caging stress in ducks. We designed this experiment to analyze the effects of caging stress on performance parameters and oxidative stress indexes in ducks. Methods: Liquid chromatography tandem mass spectrometry (LC/MS-MS) was used to determine the changes in metabolites in duck plasma at 5 (CR5), 10 (CR10), and 15 (CR15) days after cage rearing and traditional breeding (TB). The associated pathways of differentially altered metabolites were analyzed using Kyoto encyclopedia of genes and genomes (KEGG) database. Results: The results of this study indicate that caging stress decreased performance parameters, and the plasma total superoxide dismutase levels were increased in the CR10 group compared with the other groups. In addition, 1,431 metabolites were detected. Compared with the TB group, 134, 381, and 190 differentially produced metabolites were identified in the CR5, CR10, and CR15 groups, respectively. The results of principal component analysis (PCA) show that the selected components sufficiently distinguish the TB group and CR10 group. KEGG analysis results revealed that the differentially altered metabolites in duck plasma from the CR5 and TB groups were mainly associated with ovarian steroidogenesis, biosynthesis of unsaturated fatty acids, and phenylalanine metabolism. Conclusion: In this study, the production performance, blood indexes, number of metabolites and PCA were compared to determine effect of the caging stress stage on ducks. We inferred from the experimental results that caging-stressed ducks were in the sensitive phase in the first 5 days after caging, caging for approximately 10 days was an important transition phase, and then the duck continually adapted. Objective: Cage rearing has critical implications for the laying duck industry because it is convenient for feeding and management. However, caging stress is a type of chronic stress that induces maladaptation. Environmental stress responses have been extensively studied, but no detailed information is available about the comprehensive changes in plasma metabolites at different stages of caging stress in ducks. We designed this experiment to analyze the effects of caging stress on performance parameters and oxidative stress indexes in ducks.Methods: Liquid chromatography tandem mass spectrometry (LC/MS-MS) was used to determine the changes in metabolites in duck plasma at 5 (CR5), 10 (CR10), and 15 (CR15) days after cage rearing and traditional breeding (TB). The associated pathways of differentially altered metabolites were analyzed using Kyoto encyclopedia of genes and genomes (KEGG) database.Results: The results of this study indicate that caging stress decreased performance parameters, and the plasma total superoxide dismutase levels were increased in the CR10 group compared with the other groups. In addition, 1,431 metabolites were detected. Compared with the TB group, 134, 381, and 190 differentially produced metabolites were identified in the CR5, CR10, and CR15 groups, respectively. The results of principal component analysis (PCA) show that the selected components sufficiently distinguish the TB group and CR10 group. KEGG analysis results revealed that the differentially altered metabolites in duck plasma from the CR5 and TB groups were mainly associated with ovarian steroidogenesis, biosynthesis of unsaturated fatty acids, and phenylalanine metabolism.Conclusion: In this study, the production performance, blood indexes, number of metabolites and PCA were compared to determine effect of the caging stress stage on ducks. We inferred from the experimental results that caging-stressed ducks were in the sensitive phase in the first 5 days after caging, caging for approximately 10 days was an important transition phase, and then the duck continually adapted.
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Yufei Sun,Guifang Feng,Yan Zheng,Shu Liu,Yan Zhang,Zifeng Pi,Fengrui Song,Zhiqiang Liu 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.1
Background: Exploring the pharmacokinetic (PK) changes of various active components of single herbsand their combinations is necessary to elucidate the compatibility mechanism. However, the lack ofchemical standards and low concentrations of multiple active ingredients in the biological matrix restrictPK studies. Methods: A putative multiple reaction monitoring strategy based on liquid chromatography coupledwith mass spectrometry (LCeMS) was developed to extend the PK scopes of quantification withoutresorting to the use of chemical standards. First, the compounds studied, including components withavailable reference standard (ARS) and components lacking reference standard (LRS), were preclassifiedto several groups according to their chemical structures. Herb decoctions were then subjected toultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometryanalysis with appropriate collision energy (CE) in MS2 mode. Finally, multiple reaction monitoringtransitions transformed from MS2 of ultrahigh-performance liquid chromatography coupled withquadrupole time-of-flight mass spectrometry were used for ultrahigh-performance liquid chromatographycoupled with triple quadrupole mass spectrometry to obtain the mass responses of LRS components. LRS components quantification was further performed by developing an assistive groupdependentsemiquantitative method. Results: The developed method was exemplified by the comparative PK process of single herbs RadixGinseng (RG), Radix Polygala (RP), and their combinations (RGeRP). Significant changes in PK parameterswere observed before and after combination. Conclusion: Results indicated that Traditional Chinese Medicine combinations can produce synergisticeffects and diminish possible toxic effects, thereby reflecting the advantages of compatibility. The proposedstrategy can solve the quantitative problem of LRS and extend the scopes of PK studies.
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Yufei Sun,Guifang Feng,Yan Zheng,Shu Liu,Yan Zhang,Zifeng Pi,Fengrui Song,Zhiqiang Liu 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.1
Background: Exploring the pharmacokinetic (PK) changes of various active components of single herbs and their combinations is necessary to elucidate the compatibility mechanism. However, the lack of chemical standards and low concentrations of multiple active ingredients in the biological matrix restrict PK studies. Methods: A putative multiple reaction monitoring strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to extend the PK scopes of quantification without resorting to the use of chemical standards. First, the compounds studied, including components with available reference standard (ARS) and components lacking reference standard (LRS), were preclassified to several groups according to their chemical structures. Herb decoctions were then subjected to ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry analysis with appropriate collision energy (CE) in MS² mode. Finally, multiple reaction monitoring transitions transformed from MS² of ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry were used for ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry to obtain the mass responses of LRS components. LRS components quantification was further performed by developing an assistive group-dependent semiquantitative method. Results: The developed method was exemplified by the comparative PK process of single herbs Radix Ginseng (RG), Radix Polygala (RP), and their combinations (RG-RP). Significant changes in PK parameters were observed before and after combination. Conclusion: Results indicated that Traditional Chinese Medicine combinations can produce synergistic effects and diminish possible toxic effects, thereby reflecting the advantages of compatibility. The proposed strategy can solve the quantitative problem of LRS and extend the scopes of PK studies.
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Sun, Yufei,Feng, Guifang,Zheng, Yan,Liu, Shu,Zhang, Yan,Pi, Zifeng,Song, Fengrui,Liu, Zhiqiang The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.1
Background: Exploring the pharmacokinetic (PK) changes of various active components of single herbs and their combinations is necessary to elucidate the compatibility mechanism. However, the lack of chemical standards and low concentrations of multiple active ingredients in the biological matrix restrict PK studies. Methods: A putative multiple reaction monitoring strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to extend the PK scopes of quantification without resorting to the use of chemical standards. First, the compounds studied, including components with available reference standard (ARS) and components lacking reference standard (LRS), were preclassified to several groups according to their chemical structures. Herb decoctions were then subjected to ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry analysis with appropriate collision energy (CE) in MS<sup>2</sup> mode. Finally, multiple reaction monitoring transitions transformed from MS<sup>2</sup> of ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry were used for ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry to obtain the mass responses of LRS components. LRS components quantification was further performed by developing an assistive group-dependent semiquantitative method. Results: The developed method was exemplified by the comparative PK process of single herbs Radix Ginseng (RG), Radix Polygala (RP), and their combinations (RG-RP). Significant changes in PK parameters were observed before and after combination. Conclusion: Results indicated that Traditional Chinese Medicine combinations can produce synergistic effects and diminish possible toxic effects, thereby reflecting the advantages of compatibility. The proposed strategy can solve the quantitative problem of LRS and extend the scopes of PK studies.
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