Anti-inflammatory activity of chitosan nanoparticles carrying NF-κB/p65 antisense oligonucleotide in RAW264.7 macropghage stimulated by lipopolysaccharide.

The purpose of this present study is to prepare NF-κB/p65 antisense oligonucleotide loaded chitosan nanoparticles (NPs) and evaluate their physicochemical characterization and antisense effects in RAW264.7 macrophages. Condensed nanoparticles with mean particle size of 128±16nm, average Zeta potential of 19.6±6.3mV and high entrapment efficiency (EE) of 98.6±0.11% were formed between NF-κB/p65 antisense gene (NAG) and chitosan by complex coacervation method. Trypan blue staining and MTT tests showed that NAG chitosan NPs had no toxic effect on RAW264.7 macrophages when the dose was no more than 20µg/mL. Confocal microscopy images showed that NAG chitosan NPs were capable to deliver NAG into cytoplasm of RAW264.7 macrophages and finally into nucleus. Real-time PCR tests verified that NAG chitosan NPs could significantly decrease the mRNA expression level of NF-κB/p65 and inflammatory cytokines including TNF-ɑ, IL-1 and IL-6. Accordingly, western blot study showed that NAG NPs uptaken in the cells could efficiently reversed the expression of NF-κB/p65 protein induced by LPS. At last, downstream release level of inflammatory factors including TNF-ɑ, IL-1 and IL-6 in LPS stimulated RAW264.7 macrophages was significantly decreased after treated by NAG chitosan NPs. It could be concluded that chitosan NPs were excellent delivery vectors to ferry the NAG into the cytoplasm and nucleus of macrophages. The NAG chitosan NPs might be a novel therapeutic apparatus for the treatment of LPS induced sepsis by inhibiting NF-κB-related pro-inflammatory cytokines secretion.

Synthesis, characterization and DFT studies of two new pi-conjugated-conjugated pyridine-based tetrathiafulvalene derivatives.

Two new pi-conjugated pyridine-based tetrathiafulvalene derivatives, 2-(2- (4,5-bis(methylthio)-1,3-dithiol-2-ylidene)-6-phenyl-[1,3]dithiolo[4,5-b][1,4]dithiin-5-yl)pyridine (2a) and 3-(2-(4,5-bis(methylthio)-1,3-dithiol-2-ylidene)-6-(pyridin-2-yl) -[1,3]dithiolo[4,5-b][1,4]dithiin-5-yl)quinoline (2b), have been synthesized and characterized by 1H NMR, elemental analysis and mass spectroscopies. The compound 2a has also been studied by X-ray crystallography and theoretical calculations using density functional theory (DFT) framework with B3LYP/6-311+G(d,p) level of theory. Its crystal structure is triclinic system, space group P1-. The unit cell dimensions are: a = 8.813(3) Å, b = 11.082(3) Å, c = 12.620(4) Å, alfa = 88.805(5)°, beta = 80.440(5)°, gama = 75.680(5)°, V = 1177.3(6) Å3, Z = 2. The molecule exhibits one classical C-H···N intermolecular hydrogen bonds, two kinds of short intermolecular S···S interactions and two types of C-H···pi supramolecular interactions.

Fluorescence enhancement and lifetime modification of single nanodiamonds near a nanocrystalline silver surface.

Fluorescent nanodiamond (FND) contains nitrogen-vacancy defect centers as fluorophores. The intensity of its fluorescence can be significantly enhanced after deposition of the particle (35 or 140 nm in size) on a nanocrystalline Ag film without a buffer layer. The excellent photostability (i.e. neither photobleaching nor photoblinking) of the material is preserved even on the Ag film. Concurrent decrease of excited state lifetimes and increase of fluorescence intensities indicate that the enhancement results from surface plasmon resonance. Such a fluorescence enhancement effect is diminished when the individual FND particle is wrapped around by DNA molecules, as a result of an increase in the distance between the color-center emitters inside the FND and the nearby Ag nanoparticles. A fluorescence intensity enhancement up to 10-fold is observed for 35 nm FNDs, confirmed by fluorescence lifetime imaging microscopy.

The use of plasma progesterone profiles to predict the reproductive status of anestrous gilts and sows.

A plasma progesterone profile obtained from three consecutive blood samples with an interval of 7 days was evaluated for usefulness as the basis for the diagnosis and treatment of anestrous gilts and sows. Four reproductive statuses were categorized based on the plasma progesterone levels and pathological examination of the reproductive organs from 25 gilts and 12 sows with anestrus. Category 1: fluctuating (at least one sample <2.5 ng/ml and one >10 ng/ml) with normal ovary; Category 2: sustained low (<2.5 ng/ml) with inactive ovary; Category 3: persistent high (>5 ng/ml) with normal sized or cystic corpora lutea; and Category 4: animals not included in the categories mentioned, such as pigs with luteinized cysts and follicular cysts. Using the plasma progesterone profiles and this categorization, the reproductive status of 54 gilts and 38 sows with anestrus was predicted. Hormonal treatments were performed with moderate to high success. Results from this study indicate that plasma progesterone profiles can be useful for the determination of estrus status, for the diagnosis of the causes of anestrus, and for the prediction of the next estrus for an appropriate hormonal treatment in anestrous gilts and sows.