The integrated microfluidic system consists of laminar stream-based microchannels and flow cytometric analysis to enhance the human sperm motility sorting efficiency. This modified sperm sorter provided a new selection that had advances in resolving traditional sorting problems. In order to design the sorting sorting device, CFDRC simulation was performed to analyze the phenomenon of 3-D flow field. Microscopic examination revealed that sperms with different motility. For the purpose of confirming the real sorting situation, the proportions of living and dead sperm after sorting were readily identified and quantified through use of flow cytometric analysis. The viability results of sample injected from two-outlet and four-outlet chip were 95.2% and 92.2% that presented the higher quality of sorting sperm. As a result, more amounts of living sperms that mean motile sperms were counted in four-outlet channels. With this approach, the characteristics of laminar flow and a novel design were utilized to demonstrate the microfluidic system with high efficiency for sperm motility sorting. Therefore, the optimized microfluidic system provides a good opportunity to use and an inexpensive requirement to select the most appropriate sperms before IVF (in vitro fertilization) process.
Background: Airway mucus acts as an indispensable protective component of innate immune response against invading pathogens. However, airway mucus hypersecretion, largely consisting of mucin 5AC (MUC5AC), is the leading cause of airflow obstruction and airway hyperresponsiveness that contributes to chronic obstructive pulmonary disease (COPD). MicroRNAs (miRNAs) are frequently dysregulated in the pathogenesis of COPD, but the definite role of miRNAs in airway mucus hypersecretion is not well understood.Methods: A cell model of mucus hypersecretion was established in 16HBE cells by treatment with TNF-α.Cell viability and apoptosis were assessed using cell counting kit-8 (CCK-8) and flow cytometry, respectively.The aberrant expression of miR-146a-5p and miR-134-5p was assayed in TNF-α-treated 16HBE cells, and the effect of miR-146a-5p and miR-134-5p on regulating MUC5AC expression was evaluated using quantitative real-time PCR (qPCR) and Western blot analysis.Results: TNF-α treatment resulted in a significant decrease of cell viability, and increase of cell apoptosis and MUC5AC expression in 16HBE cells. Additionally, the expression of miR-134-5p and miR-146a-5p was markedly decreased in the cell model. Importantly, forced expression of miR-134-5p and miR-146a-5p significantly repressed TNF-α-induced upregulation of MUC5AC. Mechanistically, although miR-134-5p did not affect 16HBE cells viability and apoptosis, miR-134-5p partially blocked TNF-α-induced MUC5AC expression by inhibiting the activation of NF-κB signaling. On the other hand, miR-146a-5p enhanced cell viability and reduced cell apoptosis. miR-146a-5p also repressed TNF-α-induced MUC5AC expression by inhibiting p38 MAPK (mitogen-activated protein kinase) signaling activation.
Conclusions:The current data demonstrated that both miR-134-5p and miR-146a-5p conferred protection against TNF-α-induced mucus hypersecretion through repressing NF-κB and p38 MAPK signaling, indicating that miR-134-5p and miR-146a-5p may serve as the biomarker for COPD.
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