Anti-proliferative effect of the extract of Guangzao (Fructus Choerospondiatis) on cultured rat cardiac fibroblasts.

OBJECTIVE: To ascertain if total flavonoids of Guangzao (Fructus Choerospondiatis) (TFFC) extracted from Guangzao (Fructus Choerospondiatis) can inhibit angiotensin II-induced proliferation of cardiac fibroblasts (CFs). METHODS: CFs were cultured by the differential attachment method. A model of cell proliferation was established by stimulation with Ang II. Cardiac fibroblasts growth was determined using a hemocytometer. Cell proliferation was detected by methyl thiazole tetrazolium. Lactate dehydrogenase activity was measured by chemical colorimetric method. RESULTS: Proliferation of TFFC-treated (25, 50, 100 mg/L) fibroblasts was significantly less than that of cells in the angiotensin II group (P < 0.01), and TFFC inhibited proliferation in a dose-dependent manner. These inhibitory effects were partly blocked by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) and 1H-[1,2,4]-oxadiazole-[4,3-a]-quinoxalin-1-one (ODQ). CONCLUSION: TFFC inhibited angiotensin II-induced proliferation of cardiac fibroblasts via a mechanism that probably involves activation of the NO-cyclic guanosine monophosphate signaling pathway.

[In vitro effect of total flavones of Fructus Chorspondiatis on expression of collagen type I and type III mRNA and protein of cultured rat cardiac fibroblasts].

This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.

Overexpression of FcγRIIB regulates downstream protein phosphorylation and suppresses B cell activation to ameliorate systemic lupus erythematosus.

The present study aimed to examine the effects of FcγRIIB on systemic lupus erythematosus (SLE) and to investigate the underlying mechanisms. For this purpose, lentiviral vector carrying the membrane­bound type FcγRIIB gene (mFcγRIIB lentivirus) and soluble FcγRIIB (sFcγRIIB) protein were used to treat B cells from patients with SLE. The B cells were treated with calf thymus DNA (ctDNA) and anti­calf thymus DNA­immune complexes (anti­ctDNA­IC). mFcγRIIB lentivirus and sFcγRIIB protein were also injected into MRL/lpr SLE mice. The results revealed that anti­ctDNA­IC treatment significantly downregulated the IgG antibody secretion of B cells treated with mFcγRIIB lentivirus. mFcγRIIB and sFcγRIIB decreased the phosphorylation level of Bruton's tyrosine kinase (BTK) in B cells, and increased the phosphorylation level of Lyn proto­oncogene (Lyn), docking protein 1 (DOK1) and inositol polyphosphate­5­phosphatase D (SHIP). mFcγRIIB promoted the apoptosis of B cells. Following the treatment of MRL/lpr SLE mice with mFcγRIIB lentivirus, the levels of urinary protein, serum anti­nuclear and anti­dsDNA antibodies were decreased, while the levels of mFcγRIIB in B cells were increased. mFcγRIIB ameliorated the pathologies of the kidneys, liver and lymph node tissues of the MRL/lpr SLE mice. Following treatment of the MRL/lpr SLE mice with sFcγRIIB, the levels of urinary protein, serum anti­dsDNA antibody and BTK and SHIP phosphorylation levels in B cells were decreased, while the serum sFcγRIIB and sFcγRIIB­IgG levels were increased. On the whole, the findings of the present study demonstrate that recombinant FcγRIIB inhibits the secretion of IgG antibody by B cells from patients with SLE, ameliorates the symptoms of SLE in mice, and alters the phosphorylation levels of downstream proteins of the FcγRIIB signaling pathway in B cells. These results suggest that FcγRIIB may play preventive and therapeutic roles in SLE by inhibiting B cell activation via the FcγRIIB signaling pathway, which provides a novel theory and strategy for the prevention and treatment of SLE.

Bis[4-oxido-2-oxo-2,3-di-hydro-pyrimidin-1-ium-5-carboxyl-ato(1.5-)-κO,O]bis-(1,10-phenanthroline-κN,N')dysprosium(III) dihydrate.

In the title compound, [Dy(C(5)H(2.5)N(2)O(4))(2)(C(12)H(8)N(2))(2)]·2H(2)O, the Dy(III) ion is located on a twofold rotation axis and is coordinated in a square-anti-prismatic geometry by two chelating 1,10-phenanthroline mol-ecules as well as two 4-oxido-2-oxo-2,3-di-hydro-pyrimidin-1-ium-5-carboxyl-ato(1.5-) anions. N-H⋯O and O-H⋯O hydrogen bonds help to stabilize the crystal structure. The H atom involved in an N-H⋯N hydrogen bond is disordered around a twofold rotation axis.

(6-Oxido-2-oxo-1,2-dihydropyrimidine-5-carboxylato-κO,O)(4-oxido-2-oxo-1,2-dihydropyrimidin-3-ium-5-carboxyl-ato-κO,O)bis(1,10-phenanthroline-κN,N')erbium(III) dihydrate.

The erbium(III) atom in the title compound, [Er(C(5)H(2)N(2)O(4))(C(5)H(3)N(2)O(4))(C(12)H(8)N(2))(2)]·2H(2)O, is located on a twofold rotation axis and chelated by two 1,10-phenanthroline heterocycles as well as by a 2,4-dihydroxy-pyrimidine-5-car-box-yl-ate monoanion and a 2,4-dihydroxy-pyrimidine-5-car-box-yl-ate dianion in a square-anti-prismatic coordination geometry.

(µ-6-Oxido-4-oxo-1,2-dihydro-pyrimidine-5-carboxyl-ato-κO,O:O,N)bis-[aquabis-(4-oxido-2-oxo-1,2-dihydro-pyrimidin-3-ium-5-carboxyl-ato-κO,O)-(1,10-phenanthroline-κN,N')neodymium(III)] hexa-hydrate.

The water-coordinated neodymium(III) atom in the centrosymmetric title compound, [Nd(2)(C(5)H(2)N(2)O(4))(C(5)H(3)N(2)O(4))(4)(C(12)H(8)N(2))(2)(H(2)O)(2)]·6H(2)O is chelated by a 1,10-phenanthroline heterocycle and two 2,4-dihydroxy-pyrimidine-5-carboxyl-ate dianions. Two tris-chelated water-coordinated units are bridged by a 2,4-dihydroxy-pyrimidine-5-carboxyl-ate dianion, which is disordered about a center of inversion. The metal center has a monocapped square-anti-prismatic coordination.

(2,4-Dioxo-1,2,3,4-tetra-hydro-pyrimi-dine-5-carboxyl-ato-κO,O)(4-oxido-2-oxo-1,2-dihydro-pyrimidine-5-carboxyl-ato-κO,O)bis-(1,10-phenanthroline-κN,N')yttrium(III) dihydrate.

In the title compound, [Y(C(5)H(2)N(2)O(4))(C(5)H(3)N(2)O(4))(C(12)H(8)N(2))(2)]·2H(2)O, the Y(III) ion lies on a twofold rotation axis and exhibits a distorted square-anti-prismatic coordination geometry. It is chelated by two 1,10-phenanthroline ligands, a 2,4-dioxo-1,2,3,4-tetra-hydro-pyrimidine-5-carboxyl-ate mono-anion and a 4-oxido-2-oxo-1,2-dihydro-pyrimidine-5-carboxyl-ate dianion. The H atom involved in an N-H⋯N hydrogen bond between the 1,2-dihydro-pyrimidine units has half occupancy and is disordered around a twofold rotation axis.

(4-Oxido-2-oxo-1,2-dihydro-pyrimidine-5-carboxyl-ato-κO,O)(4-oxido-2-oxo-1,2-dihydro-pyrimidin-3-ium-5-carboxyl-ato-κO,O)bis-(1,10-phenanthroline-κN,N')gadolinium(III) dihydrate.

The title compound, [Gd(C(5)H(2)N(2)O(4))(C(5)H(3)N(2)O(4))(C(12)H(8)N(2))(2)]·2H(2)O, was obtained from a solvothermal reaction of 2,4-dihydroxy-pyrimidine-5-carboxylic acid (H(3)iso), GdCl(3)·6H(2)O and 1,10-phenanthroline (phen). The Gd(III) ion is located on a twofold rotation axis and is coordinated by four N atoms from two chelating phen ligands and four O atoms (5-carboxyl-ate and 4-oxido O atoms) from H(2)iso(-) and Hiso(2-) ligands. The mol-ecules are linked into a three-dimensional network by N-H⋯O, N-H⋯N and O-H⋯O hydrogen bonds. The H atom involved in an N-H⋯N hydrogen bond is disordered around a twofold rotation axis with half occupancy.

Poly[[diaquamanganese(II)]-mu(3)-4-oxido-2-oxo-1,2-dihydropyrimidine-5-carboxylato-kappa4O4,O5:O2:O2].

In the title compound, [Mn(C5H2N2O4)(H2O)2]n, the Mn(II) ion has a distorted octahedral geometry and the 4-oxido-2-oxo-1,2-dihydropyrimidine-5-carboxylate (Hiso(2-)) anion acts as a mu3:eta(4)-bridging ligand. Two oxo O atoms from different Hiso(2-) ligands bridge two Mn(II) ions, forming centrosymmetric dinuclear building blocks. Each dinuclear building block interacts with another four by the coordination of the oxide groups and carboxylate O atoms, producing a two-dimensional framework in the ab plane. Hydrogen bonds further extend the two-dimensional sheets into a three-dimensional supramolecular framework.

Quercetin-mediated synthesis of graphene oxide-silver nanoparticle nanocomposites: a suitable alternative nanotherapy for neuroblastoma.

BACKGROUND: Graphene and graphene-related materials have gained substantial interest from both academia and industry for the development of unique nanomaterials for biomedical applications. Graphene oxide (GO) and silver nanoparticles (AgNPs) are a valuable platform for the development of nanocomposites, permitting the combination of nanomaterials with different physical and chemical properties to generate novel materials with improved and effective functionalities in a single platform. Therefore, this study was conducted to synthesize a graphene oxide-silver nanoparticle (GO-AgNPs) nanocomposite using the biomolecule quercetin and evaluate the potential cytotoxicity and mechanism of GO-AgNPs in human neuroblastoma cancer cells (SH-SY5Y). METHODS: The synthesized GO-AgNPs were characterized using various analytical techniques. The potential toxicities of GO-AgNPs were evaluated using a series of biochemical and cellular assays. The expression of apoptotic and anti-apoptotic genes was measured by quantitative real-time reverse transcription polymerase chain reaction. Further, apoptosis was confirmed by caspase-9/3 activity and a terminal deoxynucleotidyl transferase dUTP nick end labeling assay, and GO-AgNPs-induced autophagy was also confirmed by transmission electron microscopy. RESULTS: The prepared GO-AgNPs exhibited significantly higher cytotoxicity toward SH-SY5Y cells than GO. GO-AgNPs induced significant cytotoxicity in SH-SY5Y cells by the loss of cell viability, inhibition of cell proliferation, increased leakage of lactate dehydrogenase, decreased level of mitochondrial membrane potential, reduced numbers of mitochondria, enhanced level of reactive oxygen species generation, increased expression of pro-apoptotic genes, and decreased expression of anti-apoptotic genes. GO-AgNPs induced caspase-9/3-dependent apoptosis via DNA fragmentation. Finally, GO-AgNPs induced accumulation of autophagosomes and autophagic vacuoles. CONCLUSION: In this study, we developed an environmentally friendly, facile, dependable, and simple method for the synthesis of GO-AgNPs nanocomposites using quercetin. The synthesized GO-AgNPs exhibited enhanced cytotoxicity compared with that of GO at very low concentrations. This study not only elucidates the potential cytotoxicity against neuroblastoma cancer cells, but also reveals the molecular mechanism of toxicity.

Polysaccharides from Trichosanthes Fructus via Ultrasound-Assisted Enzymatic Extraction Using Response Surface Methodology.

An efficient procedure for ultrasound-assisted enzymatic extraction of crude polysaccharides from Trichosanthes Fructus (crude TFP) using response surface methodology (RSM) was developed. The Box-Behnken design was applied to optimize the effects of pH (X1), enzyme amount (X2), extraction temperature (X3), and liquid-to-solid ratio (X4) on the extraction. The statistical analysis indicated that the independent variables (X4, X2, and X3), the quadratic coefficients (X12, X22, X32, and X42), and the interaction coefficient (X1X3) had significant impact on the yield of crude TFP. The optimal conditions were determined as follows: pH 4.5, enzyme amount 5000 u/g, extraction temperature 45°C, and liquid-to-solid ratio 30 ml/g. The experimental yield of crude TFP was 6.58%, which was very close to the predicted yield of 6.71%. TFPI was then purified and characterized with Sephadex G-100 column, UV-Vis, GPC, and FT-IR. The average molecular weight of TFPI was calculated to be 1.49 × 105 Da. TFPI exhibited strong reducing power and possessed not only remarkable scavenging activities against ABTS•+ and DPPH radicals, but also high antitumor activities in C4-2, DU145, and PC3 cells. The results suggest that Trichosanthes Fructus and TFPI could be a novel potent natural medicine with antioxidant and antitumor activities.

[Preparation of human soluble FcepsilonR1α and detection of serum FcepsilonR1α antibodies in patients with allergic rhinitis].

OBJECTIVE: To induce the expression of human soluble Fc epsilon receptor I alpha (sFcepsilonR1α) in a prokaryotic expression vector, purify the recombinant human sFcepsilonR1α protein, detect its binding affinity for human serum IgE antibodies and detect the levels of sFcepsilonR1α, sFcepsilonR1α-IgE and FcepsilonR1α antibodies. METHODS: The FcepsilonR1α extracellular region gene was amplified using nested polymerase chain reaction (PCR) and was expressed in a prokaryotic expression vector pET-sFcepsilonR1α using recombinant DNA technology under optimal conditions. The human sFcepsilonR1α protein was purified using iminodiacetic acid (IDA) His binding resin and identified using Western blotting. The affinity between the recombinant human sFcepsilonR1α and serum IgE antibodies and the levels of total sFcepsilonR1α, sFcepsilonR1α-IgE and FcepsilonR1α antibodies were measured using ELISA. RESULTS: The amplified gene corresponding to the extracellular region FcepsilonR1α was approximately 600 bp. PCR, double enzyme digestion and sequencing confirmed the correct sequence of the expression vector pET-sFcepsilonR1α. After human sFcepsilonR1α protein was induced in the expression vector pET-FcepsilonR1α and purified, Western blotting showed that its relative molecular mass (Mr) was approximately 42,000. ELISA revealed that the human sFcepsilonR1α bound with a high affinity to serum IgE, and the lower levels of total sFcepsilonR1α and sFcepsilonR1α-IgE and higher levels of serum anti-FcepsilonR1α antibodies in the patients with allergic rhinitis than in the normal subjects. CONCLUSION: We successfully synthesized human sFcepsilonR1α which had a strong binding affinity for human serum IgE. The higher levels of serum anti-FcepsilonR1α antibodies in the patients with allergic rhinitis than the normal subjects.

Thermally Driven Photonic Actuator Based on Silica Opal Photonic Crystal with Liquid Crystal Elastomer.

We have developed a novel thermoresponsive photonic actuator based on three-dimensional SiO2 opal photonic crystals (PCs) together with liquid crystal elastomers (LCEs). In the process of fabrication of such a photonic actuator, the LCE precursor is infiltrated into the SiO2 opal PC followed by UV light-induced photopolymerization, thereby forming the SiO2 opal PC/LCE composite film with a bilayer structure. We find that this bilayer composite film simultaneously exhibits actuation behavior as well as the photonic band gap (PBG) response to external temperature variation. When the SiO2 opal PC/LCE composite film is heated, it exhibits a considerable bending deformation, and its PBG shifts to a shorter wavelength at the same time. In addition, this actuation is quite fast, reversible, and highly repeatable. The thermoresponsive behavior of the SiO2 opal PC/LCE composite films mainly derives from the thermal-driven change of nematic order of the LCE layer which leads to the asymmetric shrinkage/expansion of the bilayer structure. These results will be of interest in designing optical actuator systems for environment-temperature detection.

Total Flavonoids of Fructus Chorspondiatis inhibits collagen synthesis of cultured rat cardiac fibroblasts induced by angiotensin II: correlated with NO/cGMP signaling pathway.

AIM: To investigate the molecular mechanism of Total Flavonoids of Fructus Chorspondiatis (TFFC) on preventing cardiac fibroblasts collagen synthesis induced by angiotensin II. METHODS: Collagen synthesis was determined by measuring (3)H-proline incorporation cardiac fibroblasts and hydroxyproline content in the culture mediums. The expression of collagen types I and III mRNA and protein was measured by RT-PCR and western blot, respectively. NO level in the culture medium was measured by the Griess reagent. NOS level in the culture medium was measured by chemical colorimetric method. The cellular concentration of cyclic GMP (cGMP) was measured by radioimmunoassay. RESULTS: TFFC (25, 50, and 100mg/L) inhibited collagen synthesis in cardiac fibroblasts in a dose-dependent manner compared with angiotensin II group (P<0.01), and the inhibitory effects were blocked by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) and 1H-[1,2,4]-oxadiazole-[4,3-a]-quinoxalin-1-one (ODQ). TFFC increased nitric oxide (NO) and nitric oxide synthase (NOS) levels in the culture medium, increased intracellular cGMP level in cardiac fibroblasts, decreased collagen types I and III protein level in cardiac fibroblasts. The mRNA expression of collagen type I and III was suppressed by TFFC. CONCLUSIONS: These results suggested that TFFC inhibited collagen synthesis induced by angiotensin II in cardiac fibroblasts, and the inhibitory effect might associate with the activation of the NO/cGMP signaling pathway.