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Background The promotion of appropriate traditional Chinese medicine (TCM) techniques in primary care institutions is an important measure emphasizing the co-development of Chinese and western medicine, and enhancing the inheritance and development of TCM. Objective To analyze primary health workers'preferences for the use of appropriate TCM techniques, providing policy recommendations for the promotion and use of such techniques in primary healthcare institutions. Methods This questionnaire survey was conducted between July and August 2021. A discrete choice model with seven attributes and 18 choice sets obtained through the use of an efficient orthogonal design were used to develop the questionnaire used in the survey, and the questionnaire has three versions each version with evenly allocated six choice sets (including an opt-out option). Additionally, the three versions have a common general demographic part (sex, age, education level, professional title, etc.) and one common consistency test question. A randomized multistage stratified cluster sampling was used to select 319 health workers on duty on the survey day from a total of 23 primary healthcare institutions in three counties in eastern, central and western China (one county was extracted from each region), and each worker was required to complete one version of the questionnaire extracted sequentially by themselves. Two hundred and ninety-five cases who returned a questionnaire passing a consistency test were included for analysis, resulting in a response rate of 92.5%. Primary health workers' preferences for appropriate TCM techniques were analyzed using conditional Logit models and latent class models. Furthermore, the changes in utility of primary health workers due to attribute level changes were monetized using willingness to pay and presented accordingly. Results The conditional Logit analysis showed that medical insurance, the average duration of each operation with the TCM technique, the influence of operation with the TCM technique on income, work burden, the promotion of professional title, and patient healthcare cost, as well as effectiveness of the TCM technique were associated with primary health workers' preferences for the use of appropriate TCM techniques (P

Introduction:Artemisia species are widely spread in north hemisphere. Artemisia sieversiana pollen is one of the common pollen allergens in the north of China. At present, seven allergens were identified and had been listed officially from A. sieversiana pollen, but the remaining allergens are still insufficiently studied, which need to be found. Methods: Pectate lyase was purified from the extracts of A. sieversiana pollen by anion exchange, size exclusion, and HPLC-hydrophobic interaction chromatography. The gene of A. sieversiana pectate lyase (Art si pectate lyase) was cloned and expressed in Escherichia coli. The enzyme activity and circular dichroism (CD) spectrum of natural and recombinant proteins were analyzed. The allergenicity of Art si pectate lyase was characterized by enzyme-linked immunosorbent assay (ELISA), Western blot, inhibition ELISA, and basophil activation test. The allergen's physicochemical properties, three-dimensional structure, sequence profiles with homologous allergens and phylogenetic tree were analyzed by in silico methods. Results: Natural Art si pectate lyase (nArt si pectate lyase) was purified from A. sieversiana pollen extracts by three chromatographic strategies. The cDNA sequence of Art si pectate lyase had a 1191-bp open reading frame encoding 396 amino acids. Both natural and recombinant pectate lyase (rArt si pectate lyase) exhibited similar CD spectrum, and nArt si pectate lyase had higher enzymatic activity. Moreover, the specific immunoglobulin E (IgE) binding rate against nArt si pectate lyase and rArt si pectate lyase was determined as 40% (6/15) in patients' serum with Artemisia species pollen allergy by ELISA. The nArt si pectate lyase and rArt si pectate lyase could inhibit 76.11% and 47.26% of IgE binding activities to the pollen extracts, respectively. Art si pectate lyase was also confirmed to activate patients' basophils. Its structure contains a predominant motif of classic parallel helical core, consisting of three parallel β-sheets, and two highly conserved features (vWiDH, RxPxxR) which may contribute to pectate lyase activity. Moreover, Art si pectate lyase shared the highest sequence identity of 73.0% with Art v 6 among currently recognized pectate lyase allergen, both were clustered into the same branch in the phylogenetic tree. Conclusion: In this study, pectate lyase was identified and comprehensively characterized as a novel allergen in A. sieversiana pollen. The findings enriched the allergen information for this pollen and promoted the development of component-resolved diagnosis and molecular therapy of A. sieversiana pollen allergy. [ABSTRACT FROM AUTHOR]