Activity and transcriptional regulatory elements of the promoter in Arctic fox (Vulpes lagopus) β-defensin103 gene / 生物工程学报

The aim of this study was to screen the active regions and transcription factor binding sites in the promoter of the CBD103 gene related to Arctic fox coat color, and to provide a basis for revealing the molecular genetic mechanism of CBD103 gene regulating the coat color formation. The 5'-flanking region fragment 2 123 bp of Arctic fox CBD103 gene was cloned, and 4 truncated promoter reporter vectors of different lengths were constructed. The promoter activity was detected by the dual-luciferase reporter assay system. Point mutations were performed on the 3 predicted specificity protein 1 (Sp1) transcription factor binding sites in the highest promoter active region, and 3 mutant vectors were constructed. The activity was then detected by the dual-luciferase reporter assay system. The results showed that the region 1 656 (-1 604/+51) had the highest activity in the 4 truncated promoters of different lengths, and the promoter activity of the three mutant vectors constructed in this region were significantly lower than that of the wild type (fragment 1 656). The region of -1 604 /+51 was the core promoter region of CBD103 gene in Arctic fox and -1 552/-1 564, -1 439/-1 454 and -329/-339 regions were positive regulatory regions. This study successfully obtained the core promoter region and positive regulation regions of the Arctic fox CBD103 gene, which laid a foundation for further study on the molecular genetic mechanism of this gene regulating Arctic fox coat color.

Clinical effect of orbital fat replacement combined with orbicularis oculi muscle flap folding for correction of severe upper eyelid depression with multiple eyelid / 中华医学美学美容杂志

Objective To explore the method and effect of orbital lateral fat replacement combined with orbicularis oculi muscle flap for correction of severe upper eyelid depression with multiple eyelid.Methods From January 2014 to January 2017,26 patients with severe upper eyelid sag with multiple eyelid were treated in Department of Plastic Surgery,Huai'an Second People's Hospital,including 18 cases of multiple physiological sag eyelid,improper operation on the formation of 8 cases of multiple eyelid eyelid sag.All the patients adopt outside orbital fat reset with the orbicularis muscle flap folded for the treatment.Results The follow-up time was 6 months to 1 year,with an average of 8.5 months.23 patients with upper eyelid sag with multiple eyelid were rectified,3 cases of upper eyelid depression associated with multiple eyelid depression alleviated,multiple eyelid disappeared,including 5 cases of preoperative patients with mild ptosis,and symptoms improved after surgery.Conclusions The orbital lateral fat replacement combined with orbicularis oculi muscle flap folding is a safe and effective treatment for upper eyelid depression with multiple eyelid.

Identification of the core promoter of the pmel gene of Bashang long-tail chickens / 生物工程学报

To explore the activity of the pmel core promoter of Bashang long-tail chickens, we constructed dual-luciferase expression vectors and transiently transfected into DF1 cells with Lipofectamine 2000. We measured the luciferase activity with the dual-luciferase detection kit. The 1 268 bp fragment in 5-flanking region of the pmel gene in Bashang long-tail chickens was cloned. The region from -1 200 bp to +68 bp included 2 CpG islands and multiple transcription factor binding sites. We constructed 9 expression vectors with different promoter regions and a mutant vector of the core promoter region of the pmel gene of Bashang long-tail chickens. The core promoter region from -840 bp to +68 bp was identified in the pmel gene. The region from -590 to -525 bp negatively regulated the pmel gene during the transcription process. The -840--590 bp and -525--266 bp regions were positive regulatory regions. The polymorphic sites (-456, -435, -410, -374 and -341) had a significant effect on the promoter activity of the pmel gene.