Co-delivery of docetaxel and p53 gene from cationic nanoparticles based on poly (l-lactide) and low-molecular-weight polyethyleneimine (PEA).

Recent findings revealed that low-concentration paclitaxel(DTX) could enhance cytotoxicity by upregulating p53 expression in lung cancer cell lines. So, co-delivery of DTX and RFP-p53 gene with PEA nanoparticles (NPs) was studied. The prepared DTX loaded PEA NPs (PEA/DTX) were characterized by particle size distribution, morphology, zeta potential, and crystallography and cytotoxicity. Results showed that the PEA/DTX NPs had a mall particle size (≤100 nm), moderate zeta potential (≥40 mV) and drug loading of 9.0%, DTX was released from PEA/DTX NPs in an extended period in vitro. More important, agarose gel electrophoresis showed that PEA/DTX cationic NPs were able to completely bind RFP-p53 gene with mean particles size and zeta potential. Studies on cellular uptake of (PEA/DTX)/RFP-p53 NPs demonstrated that both drug and gene were effectively taken up by A549 tumor cells. It was found that intravenous injection of (PEA/DTX)/RFP-p53 NPs efficiently inhibited growth of subcutaneous A549 carcinoma in vivo (p < 0.05) and was significantly less side effect than that of mice treated with the other groups. Therefore, the (PEA/DTX)/RFP-p53 NPs might be a promising candidate for A549 cancer therapy.

Enrofloxacin perturbs nitrogen transformation and assimilation in rice seedlings (Oryza sativa L.).

The extensive use of antibiotics worldwide has led to phytotoxicity and high risks to humans. Although research on the physiological toxicity of antibiotics is extensive, its influence on plant nitrogen uptake and assimilation remains unclear. The effect of enrofloxacin on nitrogen transformation and assimilation in rice (Oryza sativa L.) seedlings was investigated in this study. Enrofloxacin had no significant effect on rice growth, nitrogen assimilation and metabolism at low concentration, while significant changes were observed in high concentration. The growth of rice seedlings was inhibited, nitrate uptake was enhanced and nitrogen content increased significantly in both shoots and roots in enrofloxacin (800 µg L-1) treatment. Furthermore, enrofloxacin promoted the activity of enzymes related to nitrogen assimilation, including nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, and glutamate dehydrogenase. High enzyme activity resulted in an increase in intermediate products and protein content, suggesting that rice seedlings may detoxify enrofloxacin stress through amino acid binding and nitro-oxidative stress might be one of the reasons of phenotype change. Gas chromatography-mass spectrometry results revealed that different types of metabolites in both shoots and roots increased with enrofloxacin stress. Specifically, glycine, serine, and threonine metabolism; aminoacyl-tRNA biosynthesis; alanine, aspartate, and glutamate metabolism; butanoate metabolism; glyoxylate and dicarboxylate metabolism in shoot; and tyrosine metabolism and citrate cycle in root were affected. Moreover, a significant correlation between nitrogen content, nitrogen assimilation enzyme activity, and metabolite content was observed. Collectively, these findings reveal the potential risks of using reclaimed wastewater irrigation and/or antibiotic-containing animal fertilizers on crops.

Muse Cells Derived from Dermal Tissues Can Differentiate into Melanocytes.

The objective of the authors has been to obtain multilineage-differentiating stress-enduring cells (Muse cells) from primary cultures of dermal fibroblasts, identify their pluripotency, and detect their ability to differentiate into melanocytes. The distribution of SSEA-3-positive cells in human scalp skin was assessed by immunohistochemistry, and the distribution of Oct4, Sox2, Nanog, and SSEA-3-positive cells was determined by immunofluorescence staining. The expression levels of Sox2, Oct4, hKlf4, and Nanog mRNAs and proteins in Muse cells were determined by reverse transcription polymerase chain reaction (RT-PCR) analyses and Western blots, respectively. These Muse cells differentiated into melanocytes in differentiation medium. The SSEA-3-positive cells were scattered in the basement membrane zone and the dermis, with comparatively more in the sebaceous glands, vascular and sweat glands, as well as the outer root sheath of hair follicles, the dermal papillae, and the hair bulbs. Muse cells, which have the ability to self-renew, were obtained from scalp dermal fibroblasts by flow cytometry sorting with an anti-SSEA-3 antibody. The results of RT-PCR, Western blot, and immunofluorescence staining showed that the expression levels of Oct4, Nanog, Sox2, and Klf4 mRNAs and proteins in Muse cells were significantly different from their parental dermal fibroblasts. Muse cells differentiated into melanocytes when cultured in melanocyte differentiation medium, and the Muse cell-derived melanocytes expressed the melanocyte-specific marker HMB45. Muse cells could be obtained by flow cytometry from primary cultures of scalp dermal fibroblasts, which possessed the ability of pluripotency and self-renewal, and could differentiate into melanocytes in vitro.

Muse Cells, a New Type of Pluripotent Stem Cell Derived from Human Fibroblasts.

A new type of mesenchymal stem cells (MSCs) that expresses stage-specific embryonic antigen 3 (SSEA-3) and the mesenchymal cell marker CD105 are known as multilineage-differentiating stress-enduring (Muse) cells. Studies have shown that stem cells in suspension cultures are more likely to generate embryoid body-like stem cell spheres and maintain an undifferentiated phenotype and pluripotency. We separated Muse cells derived from human dermal fibroblasts by long-term trypsin incubation (LTT) through suspension cultures in methylcellulose. The Muse cells obtained expressed several pluripotency markers, including Nanog, Oct4, Sox2, and SSEA-3, and could differentiate in vitro into cells of the three germ layers, such as hepatocytes (endodermal), neural cells (ectodermal) and adipocytes, and osteocytes (mesodermal cells). These cells showed a low level of DNA methylation and a high nucleo-cytoplasmic ratio. Our study provides an innovative and exciting platform for exploring the potential cell-based therapy of various human diseases using Muse cells as well as their great possibility for regenerative medicine.

N,N,N,N-Tetra-ethyl-6-{2-[(E)-1-(4-nitro-phen-yl)ethyl-idene]hydrazino}-1,3,5-triazine-2,4-diamine.

The title compound, C(19)H(28)N(8)O(2), was prepared by the reaction of N(2),N(2),N(4),N(4)-tetra-ethyl-6-hydrazino-1,3,5-triazine-2,4-diamine and 1-(4-nitro-phen-yl)ethanone in ethanol at room temperature. The mol-ecular conformation is stabilized by intra-molecular C-H⋯N hydrogen-bonding inter-actions. There are also inter-molecular N-H⋯O hydrogen bonds, and C-H⋯π and π-π inter-actions, which help to stabilize the crystal structure. The centroid-centroid distance is 3.6172 (10) Šbetween adjacent benzene and 1,3,5-triazine rings.