The purpose of this study was to investigate the appropriate proportion of Caco-2 and HT29 co-culture in vitro cell models for permeability studies. The results showed that the transepithelial electrical resistance values of 9:1 and 1:0 groups (263 ± 3.61 and 300 ± 7.55) after 21-day culture were >250 Ω cm(2), which were suitable for further experiments. The confocal laser microscopy showed that the group of 9:1 (Caco-2:HT29) had the highest integrity, whereas the group of 0:1 (Caco-2:HT29) exhibited the lowest. The staining study confirmed that mucus was successfully produced by HT29 cells, and it was also produced in co-cultures with Caco-2 cells model, but the Caco-2 monocultures did not have any blue staining, which made us affirm that mucus is only produced in the presence of HT29 cells. The real-time PCR results showed that the total highest expression level of ALPi and MUC5AC was the ratio of 9:1 (Caco-2:HT29) and lowest is 1:1 (Caco-2:HT29). So we concluded that 9:1 (Caco-2:HT29) is the optimal Caco-2 to HT29 ratio in the in vitro model co-culture for permeability studies.

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How to target cancer cells with high specificity and kill cancer cells with high efficiency remains an urgent demand for anticancer drugs. Temporin-La, which belongs to the family of temporins, presents antitumor activity against many cancer cell lines. We first used a whole bioinformatic analysis method as a platform to identify new anticancer antimicrobial peptides (AMPs). On the basis of these results, we designed a temporin-La analog (temporin-Las) and related constructs containing the Arg-Gly-Asp (RGD) tripeptide, the integrin avb3 homing domain (RGD-La and RGD-Las). We detected a link between the net charges and integrin avb3 expression of cancer cell lines and the antitumor activities of these peptides. Temporin-La and its synthetic analogs inhibited cancer cell proliferation in a dose-dependent manner. Evidence was provided that the affinity between RGD-Las and tumor cell membranes was stronger than other tested peptides using a pull-down assay. Morphological changes on the cell membrane induced by temporin-La and RDG-Las, respectively, were examined by scanning electron microscopy. Additionally, time-dependent morphological changes were detected by confocal microscopy, where the binding process of RGD-Las to the cell membrane could be monitored. The results indicate that the electrostatic interaction between these cationic peptides and the anionic cell membrane is a major determinant of selective cell killing. Thus, the RGD tripeptide is a valuable ligand motif for tumor targeting, which leads to an increased anticancer efficiency by RGD-Las. These AMP-derived peptides have clinical potential as specifically targeting agents for the treatment of avb3 positive tumors.

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The effect of there fining process on the physicochemical properties, fatty acid composition, bioactive components, and antioxidant properties of evening primrose oil (EPO) was investigated. The results showed that the peroxide value, acid value, and p-anisidine value changed significantly throughout the refining process (p < .05). Twelve fatty acids were identified in oil samples, while their percentage contents were not significantly different during the refining process. Contents of tocopherol and total phenols were removed by 23.5% and 79.4%, respectively, after refining. The refining process resulted in losses of campesterol and β-sitosterol of approximately 16.0% and 13.8%, respectively. Moreover, squalene was detected in evening primrose oil for the first time, and its content decreased by 20.2% during the refining process. Furthermore, during the whole refining process, the DPPH free radical-scavenging ability of EPO decreased by 10.6%, and the FRAP reducing capacity dropped by 56%, meanwhile, its antioxidant stability was affected. How to cite this article: Pan F, Li Y, Luo X, et al. Effect of the chemical refining process on composition and oxidative stability of evening primrose oil.