RESUMO
Transcriptional regulation is crucial for regulated gene expression. Due to the complexity, it has been difficult to engineer eukaryotic transcription factor (TF) and promoter pairs. The few availabilities of eukaryotic TF and promotor pairs limit their practical use for clinical or industrial applications. Here, we report a de novo construction of synthetic inhibitory transcription factor and promoter pairs for mammalian transcriptional regulation. The design of synthetic TF was based on the fusion of DNA binding domain and Kruppel associated box transcription regulating domain (KRAB). The synthetic promoter was constructed by inserting the corresponding TF response element after SV40 promoter. We constructed and tested five synthetic inhibitory transcription factor and promoter pairs in cultured mammalian cells. The inhibition capability and orthogonality were verified by flow cytometry. In summary, we demonstrate the feasibility of constructing mammalian inhibitory TF and promoter pairs, which could be standardized for advanced gene-circuit design and various applications in the mammalian synthetic biology.
Assuntos
Animais , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Mamíferos , Regiões Promotoras Genéticas , Fatores de Transcrição , Transcrição GênicaRESUMO
Objective To investigate the effect and mechanism of autophagy inhibited by Met/HGF passway in alleviating myocardial ischemia and reperfusion injury.Methods Lsolated cardiomyocytes of rats were divided into 5 groups:control group,ischemia and reperfusion (IR) group,Met-transfection group,Met siRNA-transfection group,Met-transfection and HGF activator group.Over or less expressions of Met were controlled by the method of transfection.Cell proteins were extracted after IR injury.The expression of cell protein such as met,HGF,AMPK,PI3K,autophagy-related protein LC3B,Beclin-1 and apoptosis-related protein Bcl-2 were detected by Western blot.Results Compared with control group,Western blot results showed that the expression of AMPK and PI3K increased significantly by the method of over expression of Met and activation of HGF,the expression of autophagy-related protein LC3B and Beclin-1 decreased significantly at the same time.However,it showed an opposite trend in IR group and Met siRNA-transfection group.Conclusion Activation of the Met/HGF signal pathway inhibits autophagy and attenuates myocardial ischemia-reperfusion injury.
RESUMO
<p><b>BACKGROUND</b>The prevalence of developmental dysplasia of the hip (DDH) is unknown in China. We aimed to determine the prevalence of DDH in Chinese adults.</p><p><b>METHODS</b>In this study, we performed a cross-sectional survey of a nationally representative sample of Chinese adults. All participants underwent questionnaire investigation, physical examination, and X-ray examination. Factors associated with DDH were analyzed with logistic regression.</p><p><b>RESULTS</b>We invited 29,180 individuals aged 18 years and over to participate, randomly selected from 18 primary sampling units (street districts in urban areas and townships in rural areas). The survey and examination were completed in 25,767 people (10,296 men and 15,471 women). DDH was diagnosed in 391 people, yielding an overall DDH prevalence of 1.52%. Based on this information, we estimate the number of individuals with DDH in China to be approximately 16.05 million. DDH prevalence increased with age (odds ratio = 1.53 [1.03-2.27], P = 0.036), was significantly higher among women than men (2.07% vs. 0.75%, P< 0.001), and was higher among rural residents than urban residents (1.75% vs. 1.29%, P< 0.001). Economic development was independently associated with the presence of DDH. There was no evidence of an association between body mass index alone, education, or current smoking or drinking and risk of DDH (P > 0.05).</p><p><b>CONCLUSIONS</b>DDH has become an important public health problem. Special attention should be paid to residents with DDH. Screening for DDH should be performed in China.</p>