T-bet is a key regulator for the lineage commitment in CD4+ T helper (Th) 1 cells by activating the hallmark production of interferon-γ. Previously, two single nucleotide polymorphisms (SNPs) in the TBX21 promoter, T-1993C and T-1514C, have been shown by statistic studies to associate with systemic lupus erythematosus (SLE). The effect of -1993 SNP on the Yin Yang 1 transcription factor-mediated promoter activity has been already indicated. This study aimed to investigate roles of the T-1514C SNP on TBX21 transcription and its functional effect by luciferase reporter, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, and flow cytometric analysis of intracellular T-bet, IFN-γ, and IL-4 expression in activated CD4+ T cells. The TBX21 promoter carrying -1514C possessed significantly lower transcriptional activity than that of -1514T and was markedly downregulated by the overexpression of upstream stimulatory factor 1 (USF-1) when compared with the promoter carrying -1514T. EMSA indicated that the transcription factor USF-1 was bound to the -1514C allele probe with the affinity higher than that to the -1514T allele probe. ChIP assay suggested that USF-1 bound around -1514 of TBX21 genomic DNA in vivo in the human T cell line Jurkat with -1514C/T. The individuals carrying -1514C allele were determined to have significantly diminished expression of T-bet and IFN-γ and increased IL-4 production in CD4+ T cells compared with those of -1514T allele. The findings demonstrate that the T-1514C polymorphism affects TBX21 gene expression and Th1 cytokine production by binding USF-1 to the SNP site.
In Qinghai province, Gazella subgutturosa reginae (Adlerberg, 1931) is only distributed in Qaidam basin and it is beneficial for the balance of this ecosystem. In this paper, we present the complete mitochondrial genome of Gazella subgutturosa reginae firstly, a circularized sequence with 16,435 bp, containing a total of 13 protein coding genes, 22 transfer RNA (tRNA) genes, and 2 ribosomal RNA (rRNA) genes. The sequence is similar to other subspecies of Gazella subgutturosa, the phylogenetic tree revealed that Gazella subgutturosa reginae and Gazella subgutturosa subgutturosa are more closely related to each other. Our research is useful for the taxonomic and evolutionary research of goitered gazelle.
In order to effectively evaluate the pollution status of available lead (Pb) in farmland environment, a method for the determination of available Pb in greenhouse soil by atomic absorption spectrometry with primary secondary amine (PSA) as solid phase dispersion adsorption material was established. In combination with neutral salt (such as 0.01 mol/L CaCl2) extraction technology, neutral lead-containing aqueous solution was obtained. Based on the rapid adsorption of Pb in neutral environment and the rapid desorption of Pb under acidic conditions, the pretreatment method of sample enrichment and purification based on PSA was established by optimizing the parameters. The samples after enrichment and purification were determined by atomic absorption spectrometry. Results showed that the linearity of the method was good in the range of 0.010 ∼ 0.050 mg/kg (R2 = 0.9990). The limit of detection was 0.0051 mg/kg and the limit of quantity was 0.017 mg/kg. The recoveries were 97.6% to 99.2% and the relative standard deviation was 0.79% ∼ 1.56% at the concentrations of 0.010, 0.030 and 0.050 mg/kg.
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