Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics* ; Manganese/metabolism* ; Plants, Genetically Modified/genetics* ; Plant Proteins/physiology* ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Cation Transport Proteins/metabolism* ; Stress, Physiological

[Objective]To investigate the potential mechanism of Yuquan Capsule in treating diabetic nephropathy(DN)by network pharmacology and in vitro experiments.[Methods]The active ingredients and disease-associated genes of Yuquan Capsule were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Traditional Chinese Medicine Integrated Database(TCMID),the target genes of DN were screened by Genome Annotation Database Platform(GeneCards),Disease Gene Network(DisGeNET)database and Therapeutic Target Database(TTD).The protein-protein interaction(PPI)network was drawn by STRING 11.5 database,and visualized by Cytoscape 3.7.2 software.The key compounds and core targets were screened by CytoHubba and MCODE plugins of Cytoscape 3.7.2 software.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed by R software(ClusterProfiler package),and the molecular docking validation was performed by AutoDock and PyMOL software.[Results]There were 177 Yuquan Capsule against DN,477 active components and 135 targets of Yuquan Capsule against DN.The core targets included serine/threonine kinase 1(AKT1),interleukin-1 beta(IL1B),catalase(CAT),nitric oxide synthase 3(NOS3),leptin(LEP),insulin-like growth factor 1(IGF1)and C-C motif chemokine ligand 2(CCL2).GO enrichment analysis showed 2 557 related items,including 1 415 biological processes,657 cellular components and 48 molecular functions.KEGG pathway analysis showed that the targets of Yuquan Capsule against DN were enriched in 66 pathways,of which cyclic adenosine monophosphate(cAMP)signaling pathway and advanced glycation end products-receptor for advanced glycation end products(AGE-RAGE)signaling pathway in diabetic complications may be the key pathways of Yuquan Capsule in the treatment of DN.Molecular docking results showed that the core target and key compounds could bind well.Experimental verification showed that the protein level of CCL2 decreased in a drugdose-dependent manner,and the protein level of CAT and endothelial nitric oxide synthase(eNOs)increased in a drugdose-dependent manner.[Conclusion]Yuquan Capsule has the characteristics of multiple components and multiple targets in the treatment of DN.The mechanism may be related to the down-regulation of CCL2 and up-regulation of CAT and eNOs protein expression,inhibiting oxidative stress,and regulating the key pathway.

Objective: To explore trends in the asthma burden among Chinese children and adolescents 1-19 years old during 1990-2019. Methods: Based on data from the 2019 Global Burden of Disease Study, joinpoint regression was used to analyze the dynamic changes in the gender and age specific asthma burden, and the asthma burden in China was compared with countries that have different socio demographic indices(SDI). In addition, trends in asthma burden attributed to different risk factors were also investigated. Results: The asthma burden decreased slightly from 1990 to 1996 [annual percent change (APC)=-1.7%], then rapidly decreased from 1996 to 2005 (APC=-5.7%). The age standardized disability adjusted life years (DALYs) rate decreased from 158.55/100 000 to 88.59/100 000 in patients 1-19 years of age. From 2005 to 2017, the DALYs rate for asthma increased slowly, then rapidly. In 2017, the DALYs rate peaked at 176.18/100 000, then decreased to 126.79/100 000 in 2019. The burden of asthma in boys was higher than girls, and the DALYs rate for asthma in the group 5-9 years of age was higher than the remaining age groups. Furthermore, the age standardized DALYs rate for asthma among Chinese children and adolescents was relatively low among countries with a different SDI. In addition, the DALYs rate attributed to high body mass index increased in all age groups in China. Specifically, the average APC (AAPC) was 2.9% in group 1-4 years of age and the AAPC was 4.2% in the remaining age groups. The DALYs rate attributed to occupational asthmagens in the group 15-19 years of age decreased from 1990 to 2019 and the AAPC was -2.5%. Conclusion The asthma burden was relatively low among Chinese children and adolescents, and there were gender and age differences. The gender and age specific DALYs rate for asthma had a tendency to decrease, increase, then decrease. More attention should be paid to boys and the group 5-9 years of age, and strengthen the intervention of obesity and occupational asthmagens.