Transcriptome Analysis Reveals Key miRNA-mRNA Pathways in Ovarian Tissues of Yunshang Black Goats With Different Kidding Numbers.

The granulosa cell (GC) is the basic functional unit of follicles, and it is important for promoting follicle growth and sex hormones, as well as growth factor secretion in the process of reproduction. A variety of factors influence granulocyte proliferation, yet there are still many gaps to be filled in target and non-coding RNA regulation. In our study, the differentially expressed (DE) mRNAs and miRNAs were detected by using RNA-seq, and we constructed a mRNA-miRNA network related to goat prolificacy. Then, the goat primary GCs were isolated from the follicle for the function validation of candidate genes and their regulator miRNAs. A total of 2,968 DE mRNAs and 99 DE miRNAs were identified in the high- and low-prolificacy goat by RNA-seq, of which there were 1,553 upregulated and 1,415 downregulated mRNAs, and 80 upregulated and 19 downregulated miRNAs, respectively. JAK3 was identified as highly expressed in the low-prolificacy goats (3 times higher than high-prolificacy goats), and the integrated analysis showed that chi-miR-493-3p was a potential regulator of JAK3. The analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that JAK3 was involved in the PI3K-Akt signaling pathway, the Jak-STAT signaling pathway, and signaling pathways regulating pluripotency of stem cells. In particular, the PI3K-Akt signaling pathway was a typical pathway for cell proliferation, differentiation, apoptosis, and migration. We found that the chi-miR-493-3p targets JAK3 directly via RT-qPCR, dual fluorescence assays, and Western blot. Furthermore, the expression of JAK3 was significantly decreased by the chi-miR-493-3p mimic and increased by the chi-miR-493-3p inhibitor. The CCK-8 assay showed that overexpression of JAK3 promoted cell proliferation, while inhibiting JAK3 had the opposite effect. The expression of cell proliferation markers CDK4 and cyclin D2 also showed the same results. Moreover, the enzyme-linked immunosorbent assay showed that steroid hormones E2 and PROG were increased by overexpressing JAK3 and decreased by inhibiting JAK3. Therefore, our results identified a chi-miR-439-3p-JAK3 regulatory pathway, which provided a new insight into the GC proliferation and prolificacy of goat.

Effects of Medium Composition and Genetic Background on Agrobacterium-Mediated Transformation Efficiency of Lentinula edodes.

The establishment of genetic transformation method is crucial for the functional genomics research in filamentous fungi. Although the transformation method has been developed in several types of fungi, a highly efficient and convenient transformation system is desperately needed in Lentinula edodes. Present work established the Agrobacterium-mediated transformation (ATMT) of basidiomycete L. edodes in both monokaryon and dikaryon mycelia by using constructed binary plasmid pCAMBIA-1300-GFP. Then, the transformation efficiency of ATMT was evaluated by using different mediums for recipient incubation and different varieties of L. edodes. The results showed that in dikaryon strain W1, the positive hygromycin-resistant transformants was observed in all medium with the positive frequency of selected transformants that ranged from 0 to 30%. While in the monokaryon strain W1-26, only the millet medium group obtained positive transformants with a positive frequency of 75.48%. Moreover, three dikaryotic wild strains (YS55, YS3334, and YS3357) and two dikaryotic cultivated strains (W1 and S606) showed the highest transformation efficiency, with 32.96% of the germination frequency, and 85.12% of positive frequency for hygromycin-resistant transformants. This work demonstrated that Agrobacterium-mediated transformation was successfully performed in L. edodes, and the genotype of recipients as well as the medium for mycelial incubation were suggested to play key roles in determining the transformation efficiency. These findings may provide new avenues for the genetic modification of edible mushroom and may extend the cognition of DNA-mediated transformation in filamentous fungi.

Agrobacterium-mediated transformation of the ascomycete mushroom Morchella importuna using polyubiquitin and glyceraldehyde-3-phosphate dehydrogenase promoter-based binary vectors.

Morchella importuna is a worldwide distributed edible mushroom with high ecological and economic values, but the molecular and genetic research about this mushroom has been hindered due to lack of an efficient transformation method. Here, we report for the first time the successful transformation of M. importuna by using a hypervirulent Agrobacterium tumefaciens strain bearing the constructed binary plasmid p1391-U-GUS. The selectable markers used were the genes for hygromycin resistance under the control of the polyubquitin promoter from M. importuna. The reporter genes were those for enhanced green fluorescent protein (EGFP) and the ß-Glucuronidase (GUS) under the control of glyceraldehyde-3-phosphate dehydrogenase promoter and polyubquitin promoter respectively. The presence of the reporter gene EGFP in the transformants was confirmed by the fluorescence and confocal microscope and molecular analysis and that of the reporter gene GUS was verified by enzyme activity and molecular analysis. The analysis results of both reporter genes indicated that Agrobacterium-mediated transformation was successfully performed in M. importuna.

Sulfonated reduced graphene oxide catalyzed cyclization of hydrazides and carbon dioxide to 1,3,4-oxadiazoles under sonication.

Acid catalysts facilitate many chemical reactions. Sulfonated reduced grapheneoxide (rGOPhSO3H) has shown to be an encouraging solid acid catalyst because of its efficiency, cost-effectiveness and safety of use. In this study, we prepared the rGOPhSO3H nano acid catalyst, with the introduction of aromatic sulfonic acid radicals onto GO by fractional removal of oxygenated functions. It was thoroughly characterized by FT-IR, X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Raman spectroscopy, energy dispersive spectroscopy (EDS) and solid state 13C MAS NMR (SSNMR). Here we report the conversion of CO2 (1.0 atm pressure, at = 50 °C, the source of C1 carbon feed stock) with hydrazides and a catalytic amount rGOPhSO3H, which through a cyclization reaction results in a new strategy for the synthesis of 5-substituted-3H-[1,3,4]-oxadiazol-2-ones (SOxdOs) under ultrasonic irradiation. Hence this concept of cyclization opens up for new insights.

Water-based synthesis and processing of novel biodegradable elastomers for medical applications.

Biodegradable elastomers in the form of polyurethane nanoparticles (NPs) were successfully synthesized based on the combinations of two hydrolysis-prone polyester diols by a green water-based process. The anionic nature of the polymers successfully rendered polyurethane NPs (30-50 nm) consisting of approximately 200-300 polymer chains. The mechanical properties and degradation rate could be adjusted by the types and ratios of the component oligodiols in the soft segment. We demonstrated the feasibility using these biodegradable NPs as building blocks to generate self-assembled morphologies in nanometric, micrometric, or bulk scale, bearing excellent elasticity and biocompatibility. The elastic NPs and their various assembled forms represent a series of smart biodegradable elastomers with potential medical applications.

Organic/Metallic nanohybrids based on amphiphilic dumbbell-shaped dendrimers.

In this study, we synthesized a series of amphiphilic dumbbell-shaped dendrimers through the addition reactions of a hydrophilic poly(oxyalkylene) with hydrophobic dendrons based on 4-isocyanate-4'-(3,3-dimethyl-2,4-dioxo-azetidine)diphenylmethane with different numbers of branching generations. The addition reaction of azetidine-2,4-diones of dendrons to amines of poly(oxyalkylene) was proceeded by stirring the reactants in dry tetrahydrofuran (THF) under nitrogen at 60 °C. In aqueous media, the dumbbell-shaped dendrimers self-assembled into micelles with their hydrophobic dendrons in the core and their hydrophilic poly(oxyalkylene) segments forming loops in the corona shell. Employing the unique self-assembled micelle structures as templates for subsequent chemical reduction of the Ag(+) ions, we generated new types of organic/metallic [silver nanoparticle (AgNP)] nanohybrid clusters. The long poly(oxyalkylene) loops that extended into the aqueous phase complexed with the Ag(+) ions, providing the suspension with steric stabilization to prevent the AgNPs from collision and flocculation. After reduction, the AgNPs were present in a homogeneous distribution in the round dendrimer micelle-stabilized nanoclusters. The diameter of each AgNP was less than 10 nm; the diameter of each round nanocluster was in the range of 50-200 nm. The encapsulation efficiency of the AgNPs in micelles was about 54-69% for the dumbbell-shaped dendrimer based organic/AgNP nanohybrid.

Antibacterial properties of silver nanoparticles in three different sizes and their nanocomposites with a new waterborne polyurethane.

Silver nanoparticles (AgNPs) are strong bactericidal agents but they are also cytotoxic. Embedding them in a polymer matrix may reduce their cytotoxic effect. In the present study, AgNPs in three average sizes were tested for their antibacterial activities and cytotoxicity. Nanocomposites from a new waterborne polyetherurethane (PEU) ionomer and AgNPs were prepared without the use of any crosslinker. It was observed that the antibacterial activity of AgNPs against Escherichia coli started at the effective concentration of 0.1-1 ppm, while that against Staphylococcus aureus started at higher concentrations of 1-10 ppm. Cytotoxicity of AgNPs was observed at the concentration of 10 ppm. AgNPs with smaller average size showed greater antibacterial activity as well as cytotoxicity. The PEU synthesized in this study showed high tensile strength, and the addition of AgNPs at all sizes further increased its thermal stability. The delicate surface features of nanophases, however, were only observed in nanocomposites with either small-or medium-sized AgNPs. PEU-Ag nanocomposites had a strong bacteriostatic effect on the growth of E. coli and S. aureus. The proliferation of endothelial cells on PEU-Ag nanocomposites was enhanced, whereas the platelet adhesion was reduced. The expression of endothelial nitric oxide synthase gene was upregulated on PEU-Ag containing small-sized AgNPs (30 ppm) or medium-sized AgNPs (60 ppm). This effect was not as remarkable in nanocomposites from large-sized AgNPs. Overall, nanocomposites from the PEU and 60 ppm of the medium-sized (5 nm) AgNPs showed the best biocompatibility and antibacterial activity. Addition of smaller or larger AgNPs did not produce as substantial an effect in PEU, especially for the larger AgNPs.

Preparation and supramolecular self-assembly of amphiphilic dendron-POSS nanohybrids.

Organic/inorganic urea/malonamide dendron-POSS nanohybrids with different generations of dendrons and peripheral groups were prepared. Chemical structures of the nanohybrids were characterized with FTIR, 1H NMR, molecular mass spectrometry, and elemental analysis. The nanohybrids exhibited better solubility in organic solvents as compared to their corresponding dendrons. Self-assembled layered dendron/POSS structures were observed for the nanohybrids with the low generation dendrons, as evidenced by TEM analysis. The nanohybrids also exhibited amphiphilic property because of the hydrophilicity of urea/malonamide dendrons. Moreover, the morphologies of the nanohybrids could be tailored via using various casting solvents.

Functionalization of silica nanoparticles with 4-isocyanato-4'-(3,3'-dimethyl-2,4-dioxo-azetidino)diphenyl methane, surface chemical reactivity and nanohybrid preparation.

This paper reports a facile approach to functionalize silica nanoparticle (SNP) surfaces, using 4-isocyanato-4'-(3,3'-dimethyl-2,4-dioxo-azetidino)diphenyl methane (MIA) as a reaction agent. Incorporation of MIA onto SNP surfaces is demonstrated with infrared spectrometry, (1)H NMR, and thermal analysis. The amounts of MIA moieties bonded to SNP surfaces are adjustable with the MIA contents in reaction systems. This modification approach brings azetidine-2,4-dione groups, which are selectively reactive toward primary amines, to SNP surfaces and the result is that the MIA-modified SNPs are effective and divergent intermediates for preparation of SNP-based nanohybrid materials. Consequently, SNP-cored polystyrene star polymers and SNP-polyhedral oligomeric silsesquioxane (POSS) nanohybrids are prepared with reaction of MIA-modified SNPs with amine-functionalized agents, demonstrating the applications of MIA-modified SNP particles in nanomaterial preparations.

Orderly arranged NLO materials based on chromophore-containing dendrons on exfoliated layered templates.

Three chromophore-containing dendrons were intercalated into montmorillonite layered silicates via an ion-exchange process. Enlarged d spacings ranging from 50 to 126 A were achieved for these novel organoclays. After the organoclays were blended with a polyimide, the steric bulkiness of the dendrons and the interaction between dendron and polyimide resulted in an ordered morphology. The orderly arranged nanocomposites were characterized by a UV-visible spectrophotometer, a variable-temperature infrared spectrometer, and electro-optical modulation. The dendrons in layered silicates were capable of undergoing a critical conformational change into an ordered structure, indicated by the drastic changes of interlayer distances at certain packing densities. Electro-optical coefficients increased sharply from 0 to 6 pm/V while the conformational change occurred. Furthermore, the addition of a polyimide capable of interaction-induced orientation was found to exert an enhancing effect on the degree of the noncentrosymmetric alignment.

Stabilizing effect of pseudo-dendritic polyethylenimine on platinum nanoparticles supported on carbon.

A new type of surfactant with a hydrophile of dendritic polyethylenimine (C(12)(EI)(7)) was synthesized by a cationic polymerization of dodecylamine with aziridine and was used as a stabilizer to prepare Pt colloid, which is then used in situ to prepare carbon-supported Pt nanoparticles. The effects of supporting carbon, surfactant concentration, and calcination time on the nanoparticle size and catalytic performance were determined from the transmission electron microscopic analyses and cyclic voltammograms. In the presence of carbon, the Pt particle size increased slightly with lower C(12)(EI)(7) content, while those with higher C(12)(EI)(7) concentrations remained unchanged. For the heat-treated Pt/C catalyst, the molar ratio of C(12)(EI)(7) to H(2)PtCl(6) ([N]/[Pt] ratio) dominated the growth of Pt nanoparticles. The size decreased from 7.6 nm for a [N]/[Pt] ratio of 5 to 3.8 nm for a [N]/[Pt] ratio of 40. X-ray photoelectron spectroscopy revealed that metallic Pt(0) (81.6%) predominated the Pt species in the heat-treated catalyst, which is more than the commercial E-TEK catalyst. The data show clearly that thermal treatment had successively removed the stabilizing shells; moreover, it did not cause serious aggregation of particles in the presence of C(12)(EI)(7) and thus enhanced the catalytic activity. The interaction between Pt and C(12)(EI)(7) were studied and were explained in terms of a mechanism of dual-functional stabilization both on carbon and in the thermal treatment.

Generation and synthetic uses of stable 4-[2-isopropylidene]-phenol carbocation from bisphenol A.

[reaction: see text] Stable 4-[2-isopropylidene]-phenol carbocation, IPP cation 1, was generated readily by addition of bisphenol A in concentrated sulfuric acid at ambient temperature, and the cation could be used for facile syntheses of 4-isopropenyl phenol (IPP), IPP dimers, and spiro-bisphenol derivatives.