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

ObjectiveTo evaluate the clinical effect of Jiechang Buxu pills on chronic relapsing ulcerative colitis （syndrome of spleen-kidney Yang deficiency） and the repair effect on mucosal barrier injury. MethodA total of 108 patients were randomly assigned into observation （54 cases） and control （54 cases） groups. Both groups were treated with mesalazine enteric-coated tablets， 0.5 g/time， 3 times/d. In addition， the observation group received Jiechang Buxu pills， 10 g/time， 3 times/d， and the control group received simulant of Jiechang Buxu pills， 10 g/time， 3 times/d. Both groups were treated for 3 consecutive months. During monthly follow-up， clinical response， clinical remission rate， C-reactive protein （CRP） compliance rate， fecal calprotectin （FC） compliance rate， and endoscopic mucosal healing rate were compared. The early recurrence rate was recorded after 3 months of follow-up. The modified Mayo score， mucosal histological score， colonoscopic mucosal score， inflammatory bowel disease questionnaire （IBDQ） score， and syndrome score of spleen-kidney Yang deficiency were compared before and after treatment. The levels of hypoxia-inducible factor-1α （HIF-1α）， diamine oxidase （DAO）， D-lactic acid （D-LA）， interleukin（IL）-1β， IL-6， IL-17A， IL-22， tumor necrosis factor-α （TNF-α）， T helper 17 （Th17）， and regulatory T cells （Treg）， and Th17/Treg ratio were measured and calculated before and after treatment. ResultThe clinical response rate， clinical remission rate， and CRP compliance rate in the observation group were higher than those in the control group （P<0.05， P<0.01） 1， 2 and 3 months after treatment. The FC compliance rate in the observation group was higher than that in the control group （P<0.05） 2 and 3 months after treatment. After treatment， the observation group had higher endoscopic mucosal healing rate （P<0.01） and lower early recurrence rate （P<0.05） than the control group. In addition， the observation group had lower Mayo score， mucosal histological score， colonoscopic mucosal score， and syndrome score of spleen-kidney Yang deficiency （P<0.01）， higher IBDQ score （P<0.01）， lower levels of HIF-1α， DAO， D-LA， IL-1β， IL-6， IL-17A， TNF-α， Th17， and Th17/Treg and higher levels of IL-22 and Treg （P<0.01） than the control group. ConclusionJiechang Buxu pills combined with mesalazine enteric-coated tablets can improve the clinical response rate， clinical remission rate， CRP compliance rate， FC compliance rate， and endoscopic mucosal healing rate， reduce the early recurrence rate and disease activity， promote histological remission， and improve the quality of life in the patients with chronic relapsing ulcerative colitis （syndrome of spleen-kidney Yang deficiency）. The therapy may exert the therapeutic effect by inhibiting inflammation， inducing intestinal mucosal immune tolerance， and protecting the function of intestinal mucosal barrier.

Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.

Gitelman syndrome(GS) is an autosomal recessive genetic disease caused by mutations in the SLC12A3 gene located in chromosome 16q13. The incidence of GS is 1-10∶40 000. SLC12A3 encodes thiazide-sensitive sodium-chloride cotransporters(NCC) which play key roles in Na + and Cl - reabsorption. GS is characterized by hypokalemic metabolic alkalosis in combination with significant hypomagnesaemia and low urinary calcium excretion. There are some correlations between genotypes and phenotypes. In previous studies, more than 500 mutations have been identified and some of them have been functionally analyzed. We review genetic mutations and functional studies related to GS as well as the relationship between genotypes and phenotypes, and summarize the research process in molecular genetics of GS.