MicroRNAs (miRNAs) are increasingly implicated in regulating tumor malignance through their capacity to coordinately repress expression of tumor-related genes. Here, we show that overexpression of miR-194 in lung cancer cell lines, results in suppressing metastasis of lung cancer cells, while inhibiting its expression through 'miRNA sponge' promotes the cancer cells to metastasize. miR-194 expression is also found to be in strongly negative association with metastasis in clinical specimens of non-small cell lung cancer. We demonstrate that miR-194 directly targets both BMP1 and p27(kip1). The resulting downregulation of BMP1 leads to suppression of TGFβ activity and, thus, to downregulation of the expression of key oncogenic genes (matrix metalloproteinases MMP2 and MMP9). This leads, in turn, to decreased tumor invasion. In addition, the miRNA-194-induced suppression of p27(kip1) activates the RhoA pathway, producing enhanced development of actin stress fibers and impaired migration of cancer cells. These findings reveal two structurally independent but functionally linked branches of the regulatory and signaling pathway that together provide a bridge between the metastasis-depressing miRNA and the key genes that govern the malignancy of lung cancers.
Aim: To evaluate a flow cytometry protocol that uses reference beads for the enumeration of live and dead bacteria present in a mixture. Methods and Results: Mixtures of live and dead Escherichia coli with live:dead concentration ratios varying from 0 to 100% were prepared. These samples were stained using SYTO 9 and propidium iodide and 6-lm reference beads were added. Bacteria present in live samples were enumerated by agar plate counting. Bacteria present in dead samples were enumerated by agar plate counting before treatment with isopropanol. There is a linear relationship between the presented flow cytometry method and agar plate counts for live (R 2 = 0Á99) and dead E. coli (R 2 = 0Á93) concentrations of c. 10 4 to 10 8 bacteria per ml within mixtures of live and dead bacteria. Conclusions: Reliable enumeration of live E. coli within a mixture of both live and dead was possible for concentration ratios of above 2Á5% live and for the enumeration of dead E. coli the lower limit was c. 20% dead. Significance and Impact of the Study: The ability to obtain absolute cell concentrations is only available for selected flow cytometers, this study describes a method for accurate enumeration that is applicable to basic flow cytometers without specialized counting features. By demonstrating the application of the method to count E. coli, we raised points of consideration for using this FCM counting method and aim to lay the foundation for future work that uses similar methods for different bacterial strains.
MicroRNAs (miRNAs) have recently been recognized as targets for anti-metastatic therapy against cancer malignancy. Development of effective miRNA mediated therapies remains a challenge to both basic research and clinical practice. Here we presented the evidence for a miR-708-5p mediated replacement therapy against metastatic lung cancer. Expression of miR-708-5p was substantially reduced in metastatic lung cancer samples and cancer cell lines when compared to non-metastatic counterparts. Expression of the miRNA suppressed cell survival and metastasis in vitro through its direct target p21, and inhibited the PI3K/AKT pathway and stem cell-like characteristics of lung cancer cells. Systemic administration of this miRNA in a mouse model of NSCLC using polyethylenimine (PEI)-mediated delivery of unmodified miRNA mimics induced tumor specific apoptosis. It also effectively protected the tested animals from developing metastatic malignancy without causing any observed toxicity.The findings strongly support miR-708-5p as a novel and effective therapeutic agent against metastatic malignancy of non-small cell lung cancer.
Hybridization of silicon integrated circuits (ICs) with compound semiconductor device arrays are crucial for making functional hybrid chips, which are found to have enormous applications in many areas. Although widely used in manufacturing hybrid chips, the flip-chip technology suffers from several limitations that are difficult to overcome, especially when the demand is raised to make functional hybrid chips with higher device array density without sacrificing the chip footprint. To address those issues, Beida Jade Bird Display Limited has developed its unique wafer-level monolithic hybrid integration technology and demonstrated its advantages in making large-scale hybrid integration of functional device arrays on Si IC wafers. Active matrix micro-light-emitting diode micro-displays with a resolution of 5000+ pixel per inch were successfully fabricated using Beida Jade Bird Display Limited's monolithic hybrid integration technology. The general fabrication method is described, and the result is presented in this paper. The fabricated monochromatic micro-light-emitting diode micro-displays exhibit improved device performance than do other micro-display technologies and have great potentials in applications such as portable projectors and near-to-eye projection for augmented reality. More importantly, the wafer-scale monolithic hybrid integration technology offers a clear path for low-cost mass production of hybrid optoelectronic IC chips.
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