Cloning and catalytic analysis of Isatis indigotica chalcone isomerase in vitro / 中国中药杂志

Chalcone isomerase is a key rate-limiting enzyme in the biosynthesis of flavonoids in higher plants, which determines the production of flavonoids in plants. In this study, RNA was extracted from different parts of Isatis indigotica and reverse-transcribed into cDNA. Specific primers with enzyme restriction sites were designed, and a chalcone isomerase gene was cloned from I. indigotica, named IiCHI. IiCHI was 756 bp in length, containing a complete open reading frame and encoding 251 amino acids. Homology analysis showed that IiCHI was closely related to CHI protein of Arabidopsis thaliana and had typical active sites of chalcone isomerase. Phylogenetic tree analysis showed that IiCHI was classified into type Ⅰ CHI clade. Recombinant prokaryotic expression vector pET28a-IiCHI was constructed and purified to obtain IiCHI recombinant protein. In vitro enzymatic analysis showed that the IiCHI protein could convert naringenin chalcone into naringenin, but could not catalyze the production of liquiritigenin by isoliquiritigenin. The results of real-time quantitative polymerase chain reaction(qPCR) showed that the expression level of IiCHI in the aboveground parts was higher than that in the underground parts and the expression level was the highest in the flowers of the aboveground parts, followed by leaves and stems, and no expression was observed in the roots and rhizomes of the underground parts. This study has confirmed the function of chalcone isomerase in I. indigotica and provided references for the biosynthesis of flavonoid components.

Influence of nocturnal orthokeratology lens on sleep quality in myopic patients / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the influence of wearing nocturnal orthokeratology lens on sleep quality in myopic patients.?METHODS: Totally 30 myopic patients ( 58 eyes ) wearing nocturnal orthokeratology lens, who received medical treatment at the Department of Ophthalmology of Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine, were included in study ( treated group ) . And 30 myopic patients wearing spectacles at the same time were randomly selected as the control group. Sleep quality of two groups were assessed with Pittsburgh sleep quality index ( PSQI ) before and after wearing the lens in the first and the third months, and compared.?RESULTS:Equivalent diopter of the 30 patients wearing nocturnal orthokeratology lens was -2. 78D±0. 90D before treated with orthokeratology lens, -0. 30D ± 0. 31D after 1mo of treatment, and -0. 28D ± 0. 30D after 3mo of treatment. There was significant difference between the equivalent diopter got before treated with orthokeratology lens and that got after 1mo of treatment (P<0.05), and between the equivalent diopter got before treated with orthokeratology lens and that got after 3mo of treatment (P<0. 05). The total scores of PSQI of 30 myopic patients got before wearing nocturnal orthokeratology lens and after 1 or 3mo of that were respectively 2. 13±1. 36, 2. 47±1. 98, and 1. 74±1. 39. There only was significant difference in scores of subjective sleep quality got after 3mo of treatment between treated group and control group (P<0. 05), but no significant difference in scores of total PSQI, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, use of sleeping medication, and daytime dysfunction( P>0. 05).?CONCLUSION:Wearing nocturnal orthokeratology lens has no significant effect on the overall sleep quality index of myopic patients, but the subjective sleep quality decreased in the early stage of wearing the lens.