[Responses of chlorophyll fluorescence and non-structural carbohydrate accumulation of Castanopsis kawakamii seedlings to seed dispersal positions]. / æ ¼æ°æ ²å¹¼è‹—å¶ç»¿ç´ è§å ‰å’Œéžç»“æž„æ€§ç¢³æ°´åŒ–åˆç‰©ç§¯ç´¯å¯¹ç§å­æ•£å¸ƒä½ç½®çš„å“åº”.

When seeds fallen from the mother trees, their initial contact physical environment was litter or soil. The dispersal positions of seeds (seeds positioned on top of the litter, the soil surface and beneath the litter) determine the process of their natural regeneration. We simulated three different dispersal positions of Castanopsis kawakamii, including seeds positioned on top of the litter (2 and 4 cm litter was placed below the seed layer), soil surface (without litter), and seeds beneath the litter (2, 4, 6 and 8 cm litter covers in the upper layer of seeds). We examined the effects of seed dispersal position on the chlorophyll fluorescence characteristics, non-structural carbohydrate, specific leaf area, leaf dry matter content and nutrient content of seedlings. The results showed that leaf nitrogen content per area of seedlings had significantly positive correlation with soluble sugar content, non-structural carbohydrate content, and negative correlation with specific leaf area across different dispersal positions. Seedlings of the moderate litter cover (2 and 4 cm) adopted resource acquisitive strategies by increasing relative chlorophyll content, soluble sugar content, non-structural carbohydrate content, leaf dry matter content, leaf nitrogen content and phosphorus contents per area, and decreasing specific leaf area to achieve their demands for rapid growth. Seedlings grew on soil surface and beneath the deep litter (6 and 8 cm) adopted the resource conservative strategies with higher leaf nitrogen content per mass and specific leaf area, lower leaf dry matter content, and non-structural carbohydrate content to intercept more effective light resources to compensate for the shady environment brought by deep litter. This would further decrease the probability of seedling mortality due to 'carbon starvation'. Seedlings under litter layer stored starch in leaf, and reduced the energy consumption of photosynthetic tissues (low PSⅡ maximum photochemical efficiency) to maintain seedling growth. Comprehensive analysis of entropy method indicated that low amount of litter cover (2 cm) significantly promoted seedling growth of C. kawakamii. In the future, we could regulate the thickness of litter layer to promote the growth and regeneration of C. kawakamii seedlings in natural forest.

Bioinspired DNA nanocockleburs for targeted delivery of doxorubicin.

A variety of three-dimensional DNA assemblies have been proposed as drug carriers owing to their good biocompatibility and easy fabrication. In this study, inspired by the structure of cockleburs, a novel aptamer-tethered DNA assembly was developed for effective targeted drug delivery. The Apt-nanocockleburs were fabricated via a facile process of DNA base pairing: four complementary DNA single strands, including one aptamer-ended strand and three sticky-end strands, were applied to pair with each other. The main body of the nanocockleburs can load doxorubicin (Dox) whilst the covered aptamer spines bind to the target MCF-7 cells. The self-assembled Apt-nanocockleburs exhibit higher cell uptake as well as increased cytotoxicity to MCF-7 cells than DNA nanocockleburs without aptamers. This study provided a DNA constructing platform to produce new drug carriers with high selectivity for cancer targeted drug delivery.

Small-molecule-based generation of functional cardiomyocytes from human umbilical cord-derived induced pluripotent stem cells.

The purpose of this study was to investigate the cardiac-differentiation potential of induced pluripotent stem cells (iPSCs) generated from human umbilical cord-derived mesenchymal cells. Spontaneous beating colonies were observed at day 7 after the sequential addition of CHIR99021 and IWP-4. The combined use of CHIR99021 and IWP-4 downregulated the expression of pluripotency markers while upregulating cardiac transcription factors and cardiomyocyte-specific markers. The derived cardiomyocytes demonstrated typical sarcomeric structures and action-potential features; most importantly, the derived cells exhibited responsiveness to ß-adrenergic and muscarinic stimulations. The analyses of molecular, structural, and functional properties revealed that the derived cardiomyocytes were similar to cardiomyocytes derived from BJ foreskin fibroblast cells. In summary, our results demonstrate that functional cardiomyocytes can be generated from human umbilical cord-derived cells. The methodology described here has potential as a means for the production of functional cardiomyocytes from discarded human umbilical cord tissue.

Efficient generation of functional cardiomyocytes from human umbilical cord-derived virus-free induced pluripotent stem cells.

We have previously demonstrated that human umbilical cord-derived mesenchymal stem cells (UC-MSCs) can differentiate into cardiomyocyte-like cells. However, no contracting cells were observed during differentiation. In this study, we generated induced pluripotent stem cells (iPSCs) from UC-MSCs using mRNA reprogramming and focused on the differentiation of reprogrammed iPSCs into functional cardiomyocytes. For cardiac differentiation, the spontaneously contracting cell clusters were present on day 8 of differentiation. Immunostaining studies and cardiac-specific gene expression confirmed the cardiomyocyte phenotype of the differentiated cells. Electrophysiology studies indicated that iPSCs derived from UC-MSCs had a capacity for differentiation into nodal-, atrial-, and ventricular-like phenotypes based on action potential characteristics, and the derived cardiomyocytes exhibited responsiveness to ß-adrenergic and muscarinic stimulations. Moreover, the derived cardiomyocytes displayed spontaneous intracellular Ca2+ transients. These results demonstrate that functional cardiomyocytes can be generated from reprogrammed UC-MSCs, and the methodology described here will serve as a useful protocol to obtain functional cardiomyocytes from human mesenchymal stem cells.

MicroRNA-34a modulates the Notch signaling pathway in mice with congenital heart disease and its role in heart development.

The objective of the study was to elucidate the mechanism by which microRNA-34a (miR-34a) influences heart development and participates in the pathogenesis of congenital heart disease (CHD) by targeting NOTCH-1 through the Notch signaling pathway. Forty D7 pregnant mice were recruited for the purposes of the study and served as the CHD (n=20, successfully established as CHD model) and normal (n=20) groups. The positive expression of the NOTCH-1 protein was evaluated by means of immunohistochemistry. Embryonic endocardial cells (ECCs) were assigned into the normal, blank, negative control (NC), miR-34a mimics, miR-34a inhibitors, miR-34a inhibitors+siRNA-NOTCH-1, siRNA-NOTCH-1, miR-34a mimics+NOTCH-1 OE and miR-34a mimics+crispr/cas9 (mutant NOTCH-1) groups. The expressions of miR-34a, NOTCH-1, Jagged1, Hes1, Hey2 and Csx in cardiac tissues and ECCs were determined by both RT-qPCR and western blotting methods. MTT assay and flow cytometry were conducted for cell proliferation and apoptosis measurement. A dual luciferase reporter assay was applied to demonstrate that NOTCH-1 was the target gene of miR-34a. In comparison to the normal group, the expressions of miR-34a, Jagged1, Hes1 and Hey2 displayed up-regulated levels, while the expressions of NOTCH-1 and Csx were down-regulated in the CHD group. Compared with the blank and NC groups, the miR-34a mimics and siRNA-NOTCH-1 groups displayed reduced expressions of NOTCH-1 and Csx as well as a decreased proliferation rate, higher miR-34a, Jagged1, Hes1 and Hey2 expressions and an increased rate of apoptosis; while an reverse trend was observed in the miR-34a inhibitors group. The expressions of MiR-34a recorded increased levels in the miR-34a mimics+NOTCH-1 OE and miR-34a mimics+crispr/cas9 (mutant NOTCH-1) groups, however no changes in the expressions of NOTCH-1, Jagged1, Hes1, Hey2, Csx, as well as cell proliferation and apoptosis were observed when compared to the blank and NC groups. The results of our study demonstrated that miR-34a increases the risk of CHD through its downregulation of NOTCH-1 by modulating the Notch signaling pathway.