Effect of different drying techniques on rose (Rosa rugosa cv. Plena) proteome based on label-free quantitative proteomics.

To explore the molecular mechanisms of different processing technologies on rose tea (Rosa rugosa cv. Plena), we investigated the rose tea proteome (fresh rose tea [CS], vacuum freeze-drying rose tea [FD], and vacuum microwave rose tea [VD]) using label-free quantification proteomics (LFQ). A total of 2187 proteins were identified, with 1864, 1905, and 1660 proteins identified in CS, FD, and VD, respectively. Of those, 1500 proteins were quantified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation and enrichment analysis of differential expression proteins (DEPs) in VD vs. CS, FD vs. CS, and FD vs. VD showed that these pathways were associated with energy metabolism, the metabolic breakdown of energy substances and protein biosynthesis, such as oxidative phosphorylation, citrate cycle, carbon metabolism pathways, and ribosome and protein processing in endoplasmic reticulum. FD could ensure the synthesis of protein translation and energy metabolism, thereby maintaining the high quality of rose tea.

Isolation of mononuclear cells from CNS tissue using Percoll gradients centrifugation / 中国药理学通报

Aim To establish a rapid method to efficiently iso-late mononuclear cells from central nervous system ( CNS) tis-sues of mice that can be effectively utilized for identification of various immune cell populations in a single sample by flow cy-tometry. Methods For defining the feasibility and practicality of the method, wild-type C57BL/6 mice and two mouse models of CNS disease including EAE mice and APP/PS1 mice were used in this study. After the collection and homogenization of the brain and spinal cord tissues respectively, the mononuclear cells were isolated by spinning the 70% -30% Percoll gradients. Cell activities were detected by trypan blue staining, and the im-mune population that infiltrated CNS was identified by flow cy-tometry. Results The results of trypan blue staining showed that the survival rate of the isolated cells was above 90% in all groups. Flow cytometry analysis showed that the relative num- bers of lymphocytes infiltrating CNS of EAE and AD mice in-creased significantly compared with wild-type C57BL/6 mice. In addition, the relative numbers of Th1 and Th17 cell subsets me-diating the inflammatory response also increased significantly, while the decreased regulatory T cells frequency was observed in the two mouse models of CNS disease. Conclusions The cells isolated by the 70% ~30% Percoll gradients centrifugation can be effectively utilized for the identification of various immune cell populations in a single sample by flow cytometry. The meth-od described in this article is simple and rapid in operation and with high survival rate and activity of the cells, which can be ap-plied to the study of the mononuclear cells in CNS.