OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

Objective:To explore the will of the aged to participate in long-term care insurance in Qinhuangdao and its influencing factors under the background of aging of population.Methods:From April to May 2019, we selected 450 elderly people as subjects by stratified cluster random sampling in Qinhuangdao. All elderly people were investigated with the self-designed Long-term Care Insurance Need Questionnaire for the Aged. The individual characteristics, economic state and consciousness were analyzed with the methods of descriptive analysis, chi-square test and Logistic regression analysis.Results:A total of 445 valid questionnaires were collected. There were 60.7% (270/445) of aged with the wills to participate in long-term care insurance and 57.8% (156/270) of them hope the insurance mostly in cash or nursing service. Single factor analysis showed that there were statistical differences in the wills to participate in long-term care insurance among the aged with different ages, education levels, with or without a spouse, living situation, number of children, number of sons, primary caregivers, conscious health status, number of chronic diseases, number of hospitalizations in the past year, main source of income, monthly income, type of medical security, preference for care places (χ 2=50.228, 19.662, 26.504, 36.934, 16.456, 23.963, 44.482, 27.684, 24.783, 29.278, 22.313, 14.836, 10.446, 9.087; P<0.05) . Logistic regression analysis showed that the influencing factors of the will of the aged to participate in long-term care insurance included the ages, spouses and caregivers with statistical differences ( P<0.05) . Conclusions:Elderly people in Qinhuangdao are not willing to participate in long-term care insurance, and the elderly who are low age, have no spouse and have no caregivers are more willing to participate in long-term care insurance.