Three long-chain polyunsaturated fatty acids, docosahexaenoic acid (DHA, 22:6n-3), eicosapentaenoic acid (EPA, 20:5n-3) and arachidonic acid (ARA, 20:4n-6), are the most biologically active polyunsaturated fatty acids in the body. They are important in developing and maintaining the brain function, and in preventing and treating many diseases such as cardiovascular disease, inflammation and cancer. Although mammals can biosynthesize these long-chain polyunsaturated fatty acids, the efficiency is very low and dietary intake is needed to meet the requirement. In this study, a multiple-genes expression vector carrying mammalian A6/A5 fatty acid desaturases and multiple-genes expression vector carrying mammalian Δ6/Δ5 fatty acid desaturases and Δ6/Δ5 fatty acid elongases coding genes was used to transfect HEK293T cells, then the overexpression of the target genes was detected. GC-MS analysis shows that the biosynthesis efficiency and level of DHA, EPA and ARA were significantly increased in cells transfected with the multiple-genes expression vector. Particularly, DHA level in these cells was 2.5 times higher than in the control cells. This study indicates mammal possess a certain mechanism for suppression of high level of biosynthesis of long chain polyunsaturated fatty acids, and the overexpression of Δ6/Δ5 fatty acid desaturases and Δ6/Δ5 fatty acid elongases broke this suppression mechanism so that the level of DHA, EPA and ARA was significantly increased. This study also provides a basis for potential applications of this gene construct in transgenic animal to produce high level of these long-chain polyunsaturated fatty acid.
Acetyltransferases ; genetics ; metabolism ; Arachidonic Acid ; biosynthesis ; Docosahexaenoic Acids ; biosynthesis ; Eicosapentaenoic Acid ; biosynthesis ; Fatty Acid Desaturases ; genetics ; metabolism ; Fatty Acid Synthases ; genetics ; metabolism ; Fatty Acids, Unsaturated ; biosynthesis ; Genetic Vectors ; HEK293 Cells ; Humans ; Transfection

Objective The present study aimed to explore the effect of miR-29b on ox-LDL-stimulated vascular smooth muscle cells and the underlying mechanism.Methods VSMCs were divided into four groups:control+NC group,control+miR-29b group,ox-LDL+NC group and ox-LDL+miR-29b group.The cells were treated with 80 mg/L ox-LDL for 24 h to estab-lish the model.A cell counting Kit-8(CCK-8)assay was performed to evaluate VSMC proliferation.The expression of miR-29b,MCP-1,MCP-3,SM α-actin,SM MHC and KLF4 was detected via quantitative reverse-transcriptase polymerase chain reaction(qRT-PCR).The expression of MCP-1 and MCP-3 in the supernatant was detected via ELISA.Protein expression was exam-ined via Western blotting.A luciferase reporter assay was used to identify the relationship between KLF4 and miR-29b.Results We found that miR-29b was downregulated significantly in ox-LDL-stimulated VSMCs.The miR-29b mimics inhibited the pro-liferation and expression of KLF4,MCP-1 and MCP-3 and increased the levels of SM α-actin and SM MHC in the VSMCs trea-ted with ox-LDL.miR-29b could bind to the 3'-untranslated region(3'UTR)of KLF4 mRNA and subsequently inhibit the activi-ty of luciferase.The overexpression of KLF4 abrogated the effects of the miR-29b mimics.Conclusion In summary,miR-29b is able to regulate the proliferation and contractile markers of VSMCs via directly regulating KLF4.

How to give full play to the advantages of rehabilitation institutions, and to improve service quality and administration efficiency are some of the complex issues faced by the high-level administrators of rehabilitation institutions. This paper studied a total of 67 institutions to investigate the operation and management model of rehabilitation institutions in China, as well as their advantages and disadvantages.

The establishment of an efficient performance management system is crucial for a rehabilitation institution. This article discussed the concept of performance, performance management and ways to establish an efficient performance management system.

The structure of N-glycans on specific proteins can regulate innate and adaptive immunity via sensing environmental signals. Meanwhile, the structural diversity of N-glycans poses analytical challenges that limit the exploration of specific glycosylation functions. In this work, we used THP-1-derived macrophages as examples to show the vast potential of a N-glycan structural interpretation tool StrucGP in N-glycoproteomic analysis. The intact glycopeptides of macrophages were enriched and analyzed using mass spectrometry (MS)-based glycoproteomic approaches, followed by the large-scale mapping of site-specific glycan structures via StrucGP. Results revealed that bisected GlcNAc, core fucosylated, and sialylated glycans (e.g., HexNAc₄Hex₅Fuc₁Neu5Ac₁, N₄H₅F₁S₁) were increased in M1 and M2 macrophages, especially in the latter. The findings indicated that these structures may be closely related to macrophage polarization. In addition, a high level of glycosylated PD-L1 was observed in M1 macrophages, and the LacNAc moiety was detected at Asn-192 and Asn-200 of PD-L1, and Asn-200 contained Lewis epitopes. The precision structural interpretation of site-specific glycans and subsequent intervention of target glycoproteins and related glycosyltransferases are of great value for the development of new diagnostic and therapeutic approaches for different diseases.
Humans ; B7-H1 Antigen ; Glycosylation ; Polysaccharides/metabolism*