Genome-wide identification of miRNAs and targets associated with cell wall biosynthesis: Differential roles of dlo-miR397a and dlo-miR408-3p during early somatic embryogenesis in longan.

The dynamic alterations in cell wall (CW) biosynthesis play an essential role in physiological isolation during the plant somatic embryogenesis (SE). However, the mechanisms underlying the functions of cell wall-associated miRNAs (CW-miRNA) remain poorly understood in plant SE. Here, we have identified 36 distinct candidate miRNAs associated with CW biosynthesis from longan third-generation genome as well as miRNA transcriptome, and modified RLM-RACE validated four distinct miRNA, which specifically targeted four CW-related genes. More importantly, we found that the dlo-miR397a-antagomir significantly enhanced DlLAC7 expression and improved laccase activity. Interestingly, inhibition of dlo-miR397a increased CW lignin deposition and promoted the tightening of protodermal cell by miRNA-mimic technology during early SE. Moreover, overexpression of dlo-miR408-3p (dlo-miR408-3p-agomir) markedly decreased DlLAC12 expression. dlo-miR408-3p-agomir activated rapid cell division, thus promoting the globular embryo (GE) development, which might be due to high DNA synthesis activity in protoepidermal cells, rather than affecting lignin synthesis. The subcellular location also indicated that both DlLAC7 and DlLAC12 proteins were primarily localized in CW and regulated CW biosynthesis. Overall, our findings provided new insight on the molecular regulatory networks comprising various miRNAs associated with cell wall, and established that dlo-miR397a and dlo-miR408-3p played differential roles during early SE in longan. The findings also shed some light on the potential role of miRNA target DlLAC regulating in vivo embryonic development of plant.

Three-dimensional nanohydroxyapatite/chitosan scaffolds as potential tissue engineered periodontal tissue.

The development of suitable three-dimensional scaffold for the maintenance of cellular viability and differentiation is critical for applications in periodontal tissue engineering. In this work, different ratios of porous nanohydroxyapatite/chitosan (HA/chitosan) scaffolds are prepared through a freeze-drying process. These scaffolds are evaluated in vitro by the analysis of microscopic structure, porosity, and cytocompatibility. The expression of type I collagen and alkaline phosphatase (ALP) activity are detected with real-time polymerase chain reaction (RT-PCR). Human periodontal ligament cells (HPLCs) transfected with enhanced green fluorescence protein (EGFP) are seeded onto the scaffolds, and then these scaffolds are implanted subcutaneously into athymic mice. The results indicated that the porosity and pore diameter of the HA/chitosan scaffolds are lower than those of pure chitosan scaffold. The HA/chitosan scaffold containing 1% HA exhibited better cytocompatibility than the pure chitosan scaffold. The expression of type I collagen and ALP are up-regulated in 1% HA/chitosan scaffold. After implanted in vivo, EGFP-transfected HPLCs not only proliferate but also recruit surrounding tissue to grow in the scaffold. The degradation of the scaffold significantly decreased in the presence of HA. This study demonstrated the potential of HA/ chitosan scaffold as a good substrate candidate in periodontal tissue engineering.

Synergetic use of thermal and visible imaging techniques for contactless and unobtrusive breathing measurement.

We present a dual-mode imaging system operating on visible and long-wave infrared wavelengths for achieving the noncontact and nonobtrusive measurements of breathing rate and pattern, no matter whether the subjects use the nose and mouth simultaneously, alternately, or individually when they breathe. The improved classifiers in tandem with the biological characteristics outperformed the custom cascade classifiers using the Viola­Jones algorithm for the cross-spectrum detection of face and nose as well as mouth. In terms of breathing rate estimation, the results obtained by this system were verified to be consistent with those measured by reference method via the Bland­Altman plot with 95% limits of agreement from ? 2.998 to 2.391 and linear correlation analysis with a correlation coefficient of 0.971, indicating that this method was acceptable for the quantitative analysis of breathing. In addition, the breathing waveforms extracted by the dual-mode imaging system were basically the same as the corresponding standard breathing sequences. Since the validation experiments were conducted under challenging conditions, such as the significant positional and abrupt physiological variations, we stated that this dual-mode imaging system utilizing the respective advantages of RGB and thermal cameras was a promising breathing measurement tool for residential care and clinical applications.