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      • KCI등재

        Detection of Spinal Muscular Atrophy Using a Duplexed Real-Time PCR Approach With Locked Nucleic Acid-Modified Primers

        Pan Jianyan,Zhang Chunhua,Teng Yanling,Zeng Sijing,Chen Siyi,Liang Desheng,Li Zhuo,Wu Lingqian 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.1

        Background: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder mainly caused by homozygous deletions that include exon 7 of the survival motor neuron 1 (SMN1) gene. A nearby paralog gene, SMN2, obstructs the specific detection of SMN1. We optimized a duplexed real-time PCR approach using locked nucleic acid (LNA)-modified primers to specifically detect SMN1. Methods: An LNA-modified primer pair with 3´ ends targeting SMN1 specific sites c.835-44g and c.840C was designed, and its specificity was examined by real-time PCR and Sanger Sequencing. A duplexed real-time PCR approach for amplifying SMN1 and control gene albumin (ALB) was developed. A randomized double-blind trial with 97 fresh peripheral blood samples and 25 dried blood spots (DBS) was conducted to evaluate the clinical efficacy of the duplexed approach. This new approach was then used to screen 753 newborn DBS. Results: The LNA-modified primers exhibited enhanced specificity and 6.8% increased efficiency for SMN1 amplification, compared with conventional primers. After stabilizing the SMN1 test by optimizing the duplexed real-time PCR approach, a clinical trial validated that the sensitivity and specificity of our new approach for detecting SMA patients and carriers was 100%. Using this new approach, 15 of the screened 753 newborns were identified as carriers via DBS, while the rest were identified as normal individuals. These data reveal a carrier rate of 1.99% in Hunan province, South Central China. Conclusions: We have developed a novel, specific SMN1 detection approach utilizing real-time PCR with LNA-modified primers, which could be applied to both prenatal carrier and newborn screening.

      • KCI등재

        Polymethoxylated Flavone Extracts from Citrus Peels for Use in the Functional Food and Nutraceutical Industry

        Xiaolin Yao,Siyi Pan,Chunhong Duan,Fang Yang,Gang Fan,Xinrong Zhu,Shuzhen Yang,Xiaoyun Xu 한국식품과학회 2009 Food Science and Biotechnology Vol.18 No.5

        Polymethoxylated flavones (PMFs) extracted from Citrus sinensis ‘Jincheng’ peel were characterized by chromatographic and spectroscopic techniques. Seven individual PMF were identified. 3, 3’, 4’, 5, 6, 7-hexamethoxyflavone (HEX), nobiletin (NOB), heptamethoxyflavone (HEP), 5-demethylnobiletin (DN), and tangeretin (TAN) were characterized through electrospray ionization mass spectrometry (ESI-MS) in positive mode of protonated molecular ions [M+H]?, the diagnostic fragment ions, together with the UV-Vis spectra and high performance liquid chromatography (HPLC) elution order from literature data. Sinensetin (SIN) and tetramethyl-O-scutellarein (SCU) were isolated and identified through their MS, ¹H nuclear magnetic resonance (NMR) and UV-Vis spectral studies. The levels of PMFs in peels from different cultivars of citrus fruits grown in China were determined for the first time. The results showed that C. aurantium ‘Bitter orange’ peel was the most promising variety for HEP. C. sinensis peel was a good source for SIN and SCU.

      • KCI등재

        Control of Citrus Green and Blue Molds by Chinese Propolis

        Shuzhen Yang,Litao Peng,Yunjiang Cheng,Siyi Pan,Feng Chen 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.5

        Green and blue molds, caused by Penicillium digitatum and Penicillium italicum, respectively, are economically important postharvest diseases of citrus fruits. In this study, Chinese propolis ethyl acetate extract (PEAE) was evaluated to control P. digitatum and P. italicum on postharvest citrus fruits. The results indicated PEAE strongly inhibited mycelia growth and induced hyphae prominent abnormal morphological alterations. Also, PEAE had strong detrimental effect on spore germination of the tested pathogens in a concentrationdependent manner. For in vivo tests, PEAE could both reduce decay caused by P. digitatum and P. italicum respectively in wound-inoculated fruit and naturally infected fruit; meanwhile, no negative influences on the overall quality of the citrus fruits were observed with PEAE treatment. Therefore, PEAE could be used as a natural antifungal agent to control citrus blue and green mold.

      • KCI등재

        Volatile Compounds of Orange Wines Produced with and without Peel Contact

        Gang Fan,Xiaolin Yao,Yongxia Xu,Huanhuan Li,Hongfei Fu,Kexing Wang,Siyi Pan 한국식품과학회 2009 Food Science and Biotechnology Vol.18 No.6

        The present study focused on the effects of peel contact on the volatile compounds of orange wines. The volatile compounds were analyzed by sensory and instrumental analyses. Solid-phase microextraction (SPME) was used for extraction of volatile compounds. A total of 19 and 27 volatile compounds were identified in without and with peel contact wines respectively. Esters were quantitatively the dominant group of volatile compounds in without peel contact wines, while terpenes were the most abundant compounds in peel contact wines. Totally 11 and 14 new formed compounds were found in without and with peel contact wines, mainly were esters, alcohols, and acids. According to sensory analysis, the peel contact wine showed a more citrus-like and fruity aroma than the wines without peel contact.

      • KCI등재

        Antifungal Activity and Action Mode of Pinocembrin from Propolis against Penicillium italicum

        Litao Peng,Shuzhen Yang,Yunjiang Cheng,Feng Chen,Siyi Pan,Gang Fan 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.6

        The antifungal activity and possible mode of action of pinocembrin isolated from propolis against Penicillium italicum were investigated. Pinocembrin exhibited strong antifungal activity against P. italicum in a dosedependent manner. Respiration rates of P. italicum during spore germination and mycelial growth were significantly inhibited when exposure to pinocembrin. The respirations of mitochondria in state 2 and state 3 from mycelia were significantly inhibited in the presence of this compound. The phosphorylated adenosine nucleotides levels in hyphae of P. italicum treated with pinocembrin were significantly low and energy charge value became unstable. Unltrastructure of hyphae was seriously damaged with pinocembrin incubation for 24 h, which was further confirmed by the increase of relative ionic leakage and soluble protein loss of P. italicum mycelia treated with pinocembrin. It was concluded that pinocembrin inhibited the mycelial growth of P. italicum by interfering energy homeostasis and cell membrane damage of the pathogen. Pinocembrin would be a promising bioactive compound for treatment of P. italicum infections on postharvest citrus fruit.

      • KCI등재

        Study on the optimization of the decolorization of orange essential oil

        Jing-Nan Ren,Yan Zhang,Gang Fan,Mei-Ping Wang,Lu-Lu Zhang,Zi-Yu Yang,Siyi Pan 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.4

        The effects of diatomite, activated clay and acticarbon on the decolorization of orange essential oil were investigated. Single factor and orthogonal tests were performed to determine the optimum discoloring conditions. The results showed that the activated clay exhibited the most satisfactory effect on discoloring. Then it was used as the decolorizer for the decolorization of orange essential oil. The highest decolorization rate (84.5%) was obtained using 10% activated clay at 60 C for 30 min. The contents of oxygenated compounds (linalool and citral) increased from 1.4 to 3.1% after decolorization. Sensory assessment revealed that the orange essential oil after decolorization using activated clay had a mellow and characteristic orange aroma. Chromaticity analysis showed that it had excellent transparency and yellow color under the optimized condition. Thus, decolorization with activated clay could maintain the quality and prolong the storage of orange essential oil.

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