Aptamer-cocktail Functionalized Nano-microfluidic Chip for Enhancing Isolation and Characterization of Circulating Cancer Cells.

BACKGROUND/AIM: Circulating tumor cells (CTCs) have been shown to have a correlation to metastasis and prognosis of patients with cancer. The enumeration and downstream gene analysis of CTCs have attracted efforts for personalized medicine. However, enumeration and phenotypic profiling in most capture devices are challenging due to the rarity and heterogeneity of CTCs. Here, we report an aptamer-cocktail strategy coupled in nano-microfluidic chip for enhancing isolation performance and characterizing the phenotypes of CTCs. MATERIALS AND METHODS: Aptamer-cocktail recognizing EpCAM/Vimentin/EGFR/CD44 were bound to a nanopillar array on a nano-microfluidic chip. The recognition was validated with cancer cells by flow cytometry and the critical parameters were optimized with the nano-microfluidic chip. Finally, the system was applied to clinical samples. RESULTS: The proposed aptamer-cocktail showed the predominant affinity with MDA-MB-231 and SK-BR-3. When utilized to capture artificial clinical samples, it showed 71% to 83% capture efficiency. CTC detection rate was 100% in five pre-treatment and five post-treatment breast cancer patients. The enumeration data ranged from 6-33 per 2 ml. The number of CTCs in breast cancer patients before therapy was 1.27-2 times higher than that after therapy. CTCs with epithelial and mesenchymal phenotype were both detected and identified; interestingly, the mean diameter of CTCpck pos acquired in these cases was much larger than that of CTCvimentin pos Conclusion: The nano-microfluidic chip not only made it easier to phenotyping epithelial-like or mesenchymal-like CTCs, but can also be used to detect downstream genetic variation. The established platform can be applied in clinical research and facilitate auxiliary diagnosis with tumor recurrence and metastasis in advance.

Application of SmartRoot in Phenotype Profile Analysis of Armillaria gallica / 中国实验方剂学杂志

Objective:To establish a research platform for obtaining accurate phenotypic spectrum data is a technical difficulty that needs to be resolved in the research of traditional Chinese medicine resources. For example，the traditional phenotypic characterization method of Armillaria rhizomorph is mostly in a form of descriptive text，which is subjective and empirical. There is an urgent need for an objective and accurate method to characterize the phenotype of honey fungus rhizomorph. Method:Based on the image processing software Image J and the root identification plug-in SmartRoot combined with the Synbiosis ProtoCol 3 image analyzer，the growth picture of Armillaria spp. was analyzed，and the length，growth rate，branching situation，and angle of nascent rhizomorph of Armillaria gallica were measured to establish a measurement system for the phenotypic analysis of Armillaria rhizomorph. Result:Based on the method developed in this paper，the growth length，growth rate，number of branches，angle of nascent rhizomorph，and other phenotypic changes can be analyzed in a real-time manner without affecting the growth of Armillaria gallica. Armillaria spp. grew fastest at 9-12 days after generation，and the angle between the nascent rhizomorph and the parent rhizomorph was nearly vertical. This method had a certain correlation with the dry weight of traditional Armillaria biomass phenotypic parameters，with a high value in practical application. Conclusion:This study has established an objective，accurate，fast and real-time phenotypic analysis and measurement system for Armillaria rhizomorph，which expands the scope of application of SmartRoot and can be used for phenotypic analysis of traditional Chinese medicine resources under controlled experimental conditions.

Effect of olive oil phenolic compounds on osteoblast differentiation.

BACKGROUND: Osteoporosis is a skeletal disorder characterized by compromised bone strength that predisposes individuals to an increased risk of fracture. Previous in vivo and in vitro studies have reported that phenolic compounds present in extra virgin olive oil have a beneficial effect on osteoblasts in terms of increase cell proliferation. The aim of this study was to determine whether phenolic compounds present in olive oil could modify the expression of cell differentiation markers on osteoblasts. STUDY DESIGN: An in vitro experimental design was performed using MG-63 osteoblasts cell line. METHODS: MG63 cells were exposed to different doses of luteolin, apigenin, or p-coumaric, caffeic or ferulic acid. Alkaline phosphatase (ALP) was evaluated by spectrophotometry and antigen expression (cluster of differentiation [CD] 54, CD80, CD86 and HLA-DR) by flow cytometry. RESULTS: At 24 hour, treated groups showed an increased ALP and modulated antigen profile, with respect to the nontreated group. CONCLUSION: These results demonstrate that the phenolic compounds studied induce cell maturation in vitro, increasing ALP synthesis and reducing the expression of antigens involved in immune functions of the osteoblast which would improve bone density.