Downregulation of microRNA-9 reduces inflammatory response and fibroblast proliferation in mice with idiopathic pulmonary fibrosis through the ANO1-mediated TGF-ß-Smad3 pathway.

Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease with increasing occurrence, high death rates and unfavorable treatment regimens. In the current study, we identified the expression of microRNA-9 (miR-9) and anoctamin-1 (ANO1) in IPF mouse models induced by bleomycin, and their effects on inflammation and fibroblast proliferation through the transforming growth factor-ß (TGF-ß)-Smad3 pathway. To verify the targeting relationship between miR-9 and ANO1, we used bioinformatics prediction and conducted a dual-luciferase reporter gene assay. The underlying regulatory mechanisms of miR-9 and the target gene ANO1 were investigated mainly with the treatment of miR-9 mimic, miR-9 inhibitor, or siRNA against ANO1 in fibroblasts isolated from IPF mice. Enzyme-linked immunosorbent assay was performed to investigate the effect of miR-9 or ANO1 on inflammatory factors. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry were used to detect fibroblast proliferation and apoptosis. Reverse transcription quantitative polymerase chain reaction and western blot analysis were applied to measure the expression of the TGF-ß-Smad3 pathway-related genes. The determination of luciferase activity suggested that miR-9 targets ANO1. Upregulation of miR-9 or silencing of ANO1 intensified inflammation in IPF, promoted proliferation and inhibited apoptotic ability of lung fibroblasts. MiR-9 negatively modulated ANO1, and thus activated the TGF-ß-Smad3 pathway. These findings suggest that miR-9 can indirectly activate the TGF-ß-Smad3 pathway by inhibiting the expression of ANO1, thereby aggravating inflammation, promotes proliferation and suppressing apoptosis of lung fibroblasts in mice models of IPF.

The growth of ZnO on stainless steel foils by MOCVD and its application in light emitting devices.

Direct fabrication of semiconductor light emitting devices on metal foils is beneficial, because it brings flexibility and good heat sink in the devices. In this work, we have grown ZnO on the commercially available stainless steel foils by metal-organic chemical vapor deposition for the first time. With the increase of growth temperature, the morphology changes from a thin film structure to closely stacked columns, and eventually to nanorods. The change in the migration ability of adatoms due to the increase of growth temperature plays an important role in the evolution of morphology. The samples with nanorod morphology exhibit relatively better crystallinity and optical quality. A PEDOT: PSS/PMMA/ZnO device was fabricated based on the grown ZnO nanorods. The metal-insulator-semiconductor type device shows an uncommon symmetric I-V curve. Under reverse bias, the device emits fairly pure UV light, which comes from the near band edge emission of ZnO. The working mechanism of the devices has been discussed, and a model mainly based on the Poole-Frenkel effect is proposed to describe the charge transportation of the devices.

[Synthesis and Properties of 1,11,15,25-Tetrahydroxy-4,8,18,22-Di (Bridged Dipropionate Carboxyl) Phthalocyanine Copper].

In this dissertation, we study the synthesis and character of new substituted Phthalocyanine. Due to the widely application of Pcs in the fields, such as the communication, medical treatment, chemical industry and so on, therefore, they have been a hot topic over several decades by scientists. Nowadays, scientists have prepared thousands of Pcs and their derivatives. However, along with the human society development and the progress in science and technology, the new phthalocyanine with novle characteristics are still the goal of the scientists. In this dissertion, the synthetic methods of the phthlocyanine is improved. The synthesis and characterization of 1,11,15,25-tetrahydroxy-4,8,18,22-di(bridged dipropionate carboxyl) phthalocyanines are reported in this paper. The mixtures of malonic acid and 3,6-dihydroxy-phthalonitrile was added to water under stiriing. Then, a catalyst amount of sulfuric acid was added. The first synthetic precursor, i. e., malonic acid 3,3'-bis(6-hydroxy phthalonitrile) butter, its molecular formula is C19H8N4O6. phthalocyanines was prepared by malonic acid 3,3'-bis(6-hydroxy phthalonitrile) butter and dihydrate zinc acetate, copper acetate monohydrate in n-amyl alcohol, using DBU as a catalyst under the 135 °C, molecular formula of phthalocyanine complexes is C38H16N8O12M. The product was characterized by Ultraviolet-visible (UV/Vis) Spectrum absorption and fluorescence, The results are agreement with the proposed structures. And electrochemical properties were studied.

[Study on spectra properties of novel octa-substituted phthalocyanines].

The spectrum properties of four novel 1, 4, 8, 11, 15, 18, 22, 25-octaoxybutyl copper phthalocyanine; 1,4,8,11,15,18, 22, 25-octamethoxybutanoate manganese phthalocyanine; 1, 4, 8, 11, 15, 18, 22, 25-octamethoxybutanoate copper phthalocyanine; 1, 4, 8, 11, 15, 18, 22, 25-octamethoxybutanoate zinc phthalocyanine were investigated by infrared, fluorescence and UV-visible spectrum in the the paper. There is no rule in the infrared spectrum of these octa-substituted phthalocyanines. The orders of the Q band, B band and Pc dimer band are different among the above Octa-substituted Phthalocyanines in the UV and fluorescence spectra. The reason is related to the interaction between the ligand and the central metal of these octa-substituted phthalocyanines.

[Electroluminescence character of novel unsymmetry substituted phthalocyanines].

The authors for the first time fabricated OLEDs employing novel phthalocynines: 2(3)-(p-tert-butylphenoxy) copper phthalocyanine(1), 2(3),16(17)-di(p-tert-butyl-phenoxy) copper phthalocyanine(2) and 2(3), 9(10), 16(17)-tri (p-tert-butylphenoxy) copper phthalocyanine(3) as light emitting layer, and their electroluminescence character was studied. The final structures of three-layer OLEDs based on copper 2(3)-(p-tert-butylphenoxy) copper phthalocyanine (1) and 2(3), 9(10), 16(17)-tri (p-tert-butylphenoxy) copper phthalocyanine(3) were ITO/NPB(40 nm)/Pc(30 nm)/AlQ(43.5 nm)/LiF (0.5 nm)/Al(120 nm). The structure of three-layer OLED based on 2(3), 9(10), 16(17)-tri (p-tert-butylphenoxy) copper phthalocyanine (3) was ITO/NPB(30 nm)/Pc(30 nm) /BCP(20 nm)/A1Q(30 nm)/LiF (0. 5 nm)/Al(120 nm). Room-temperature electroluminescence was observed at about 869 nmand 1 062 nm for 2(3)-(p-tert-butylphenoxy) copper phthalocyanine(1); room-temperature electroluminescence of 2(3),16(17) -di(p-tert-butyl-phenoxy) copper phthalocyanine(2) was found at about 1050 nm and 1110 nm; and room-temperature electroluminescence of 2(3), 9(10), 16( 17)-tri (p-tert-butylphenoxy) copper phthalocyanine(3) was studied at about 1095 and 1204 nm. The emission wavelengths and the half bandwidths were quite different for the phthalocyanine, which may be due to the differences in the number of substituted and the molecular aggregations in vacuum sublimed films. The difference in Stokes shift relaxation was also induced by the molecular aggregations.

[Effect of peroxisome proliferators activated receptor gamma and its ligand on airway mucus hypersecretion in rats].

OBJECTIVE: To explore the effect and the molecular mechanisms of peroxisome proliferators activated receptor gamma (PPAR-gamma) and its ligand on airway mucus hypersecretion. METHODS: Thirty-six Sprague-Dawley rats were randomized into the following groups: (1) Rats in the saline control group (n = 6) received normal saline inhalation; (2) Rats in the rosiglitazone control group (n = 6) received inhaled saline and oral rosiglitazone 8 mg/kg simultaneously; (3) Rats in the acrolein group (n = 6) received inhaled acronine 3.0 mg/L, 6 h/day, for 12 days; (4) Rats in the rosiglitazone intervention group (n = 18) received inhaled acrolein and oral rosiglitazone 2 mg/kg, 4 mg/kg, 8 mg/kg, respectively, as the low dose, the moderate dose and the high dose intervention groups (n = 6 each). The lung tissue sections were stained with HE for histopathological examination. The changes of airway mucus were examined with AB-PAS. Expressions of MUC5AC and PPAR-gamma protein in the bronchial epithelium were detected by immunohistochemistry. The expression of mRNA was measured with real time RT-PCR. The data were analyzed with SPSS 10.0 software. Variables were compared with One-Way ANOVA and q test. The correlations between variables were analyzed using Pearson's correlation coefficient. RESULTS: The levels of airway mucus were (60.2 +/- 9.3)%, (4.9 +/- 1.0)%, (53.3 +/- 8.5)%, (26.5 +/- 7.4)%, (12.5 +/- 3.7)% respectively in the acrolein group, the saline control group, the low dose rosiglitazone intervention group, the moderate dose rosiglitazone intervention group, and the high dose rosiglitazone intervention group, the difference being significant among groups (F = 93.80, P < 0.01). The protein expressions of MUC5AC in the bronchial epithelium examined by immunohistochemistry were 4339 +/- 453, 1636 +/- 282, 3996 +/- 346, 3048 +/- 331, 2376 +/- 343 respectively in the acrolein group, the saline control group, the low dose rosiglitazone intervention group, the moderate dose rosiglitazone intervention group, and the high dose rosiglitazone intervention group, the difference being significant among groups (F = 67.74, P < 0.01). The protein expressions of PPAR-gamma were 1159 +/- 184, 838 +/- 151, 1272 +/- 189, 1568 +/- 282, 1872 +/- 270 respectively in the acrolein group, the saline control group, the low dose rosiglitazone intervention group, the moderate dose rosiglitazone intervention group, and the high dose rosiglitazone intervention group, the difference being significant among groups (F = 21.53, P < 0.01). The mRNA expressions of MUC5AC (the relative copies) were 35.3 +/- 10.0, 2.2 +/- 0.7, 30.5 +/- 10.2, 18.6 +/- 5.3, 10.8 +/- 2.6 respectively in the acrolein group, the saline control group, the low dose rosiglitazone intervention group, the moderate dose rosiglitazone intervention group, and the high dose rosiglitazone intervention group, the difference being significant among groups (F = 29.67, P < 0.01). The mRNA expressions of PPAR-gamma (the relative copies) were 7.8 +/- 1.9, 2.0 +/- 0.6, 9.8 +/- 2.8, 18.6 +/- 5.3, 31.6 +/- 8.9 in the acrolein group, the saline control group, the low dose rosiglitazone intervention group, the moderate dose rosiglitazone intervention group, and the high dose rosiglitazone intervention group, the difference being significant among groups (F = 39.47, P < 0.01). The expression of MUC5AC mRNA was negatively correlated with the protein expression of PPAR-gamma in the acrolein group (r = -0.880, P < 0.01). CONCLUSIONS: PPAR-gamma was involved in airway mucus hypersecretion induced by acrolein. PPAR-gamma and its ligand rosiglitazone inhibited acrolein-induced airway mucus hypersecretion, possibly through downregulation of MUC5AC.

VEGFR-2 antagonist SU5416 attenuates bleomycin-induced pulmonary fibrosis in mice.

OBJECTIVE: Abnormal angiogenesis is a central hallmark for the development and progression of idiopathic pulmonary fibrosis. It has been shown that vascular endothelial growth factor (VEGF) is one of the critical angiogenic factors in angiogenesis. The aim of the present study was to assess whether disruption of VEGF pathway would attenuate bleomycin-induced pulmonary fibrosis. METHODS: Bleomycin-induced pulmonary fibrosis mice were treated intraperitoneally with VEGF receptor tyrosine kinase inhibitor SU5416 at different phases after bleomycin infusion. We measured angiogenesis and inflammatory response in both bleomycin-treated and control mice, and correlated these levels with pulmonary fibrosis. RESULTS: The increased expressions of VEGF/VEGFR (Flk-1) were correlated to a larger number of microvessels and a higher score of pulmonary fibrosis. Early administration of SU5416 inhibited pulmonary collagen deposition, histopathologic fibroplasias and the activation of TGF-beta1/Smad3 signaling pathway in bleomycin-stimulated lung. These were also paralleled by a reduction of VEGF/VEGFR-2 (Flk-1) expression and microvessel numbers in lung. Furthermore, SU5416 inhibited inflammatory cell numbers and LDH activity in BALF and IL-13 expression in lung tissue at early inflammatory phase of bleomycin-induced pulmonary fibrosis. CONCLUSION: These results suggest that the VEGFR-2 inhibitor, SU5416, attenuates histopathologic fibroplasias and collagen deposition by regulating angiogenesis and inflammation in the lung.

2-Phenyl-imidazolium nitrate monohydrate.

In the title hydrated mol-ecular salt, C(9)H(9)N(2) (+)·NO(3) (-)·H(2)O, the dihedral angle between the aromatic rings in the cation is 11.09 (8)°. In the crystal, the components are linked into chains propagating in [101] by N-H⋯O and O-H⋯O hydrogen bonds.

Bis(2-phenyl-1H-imidazole-κN)silver(I) nitrate.

The asymmetric unit of the title compound, [Ag(C(9)H(8)N(2))(2)]NO(3), contains one complete [Ag(C(9)H(8)N(2))(2)](+) cation and two half-cations (with the other halves generated through inversion) and two NO(3) (-) anions. Each Ag(I) ion shows a linear AgN(2) coordination. The ions are linked by N-H⋯O hydrogen bonds.

Protein phosphatase-2A is a target of epigallocatechin-3-gallate and modulates p53-Bak apoptotic pathway.

(-)-Epigallocatechin-3-gallate (EGCG) is a well-known chemoprevention factor. Recent studies have revealed that EGCG triggers cancer cells undergoing apoptosis through p53-dependent pathway. How EGCG activates p53-dependent apoptosis is not fully understood. In the present study using JB6 cell as a model system, we have shown that EGCG can negatively regulate protein serine/threonine phosphatase-2A (PP-2A) to positively regulate p53-dependent apoptosis. First, EGCG at physiologic levels down-regulates PP-2A at the protein and enzyme activity levels. Second, EGCG induces apoptosis of JB6 cells, which is associated with hyperphosphorylation of p53 and up-regulation of the proapoptotic gene, Bak. DNA sequence analysis, gel mobility shifting, chromatin immunoprecipitation, and reporter gene activity assays revealed that p53 directly controls Bak in JB6 cells. Knockdown of p53 and Bak expression with RNAi substantially inhibits EGCG-induced apoptosis. Third, PP-2A directly interacts with p53 and dephosphorylates p53 at Ser-15 in vitro and in vivo. Fourth, overexpression of the catalytic subunit for PP-2A down-regulates p53 phosphorylation at Ser15, attenuates expression of the downstream proapoptotic gene, Bak, and antagonizes EGCG-induced apoptosis. Inhibition of PP-2A activity enhances p53 phosphorylation at Ser-15 and up-regulates Bak expression to promote EGCG-induced apoptosis. Finally, in the p53(-/-) H1299 and p53(+/+) H1080 cells, EGCG down-regulates PP-2A similarly but induces differential apoptosis. In summary, our results show that (a) PP-2A directly dephosphorylates p53 at Ser-15; (b) P53 directly controls Bak expression; and (c) EGCG negatively regulates PP-2A. Together, our results show that EGCG-mediated negative regulation of PP-2A is an important molecular event for the activation of p53-dependent apoptosis during its chemoprevention.

Hormonally-regulated differential expression of the two duplicated insecticyanin genes, ins-a and ins-b, during development of the tobacco hornworm, Manduca sexta.

The effects of juvenile hormone (JH) and 20-hydroxyecdysone (20E) on the developmental expression of the two insecticyanin genes, ins-a and ins-b, were investigated with two gene-specific probes. Removal of the corpora allata (-CA, source of JH) clearly delayed and down-regulated the epidermal expression of these genes but enhanced their expression in the fat body during the early development of the fifth instar. Application of JH I to the -CA larvae at the time of head capsule slippage completely restored the normal epidermal expression pattern of the two genes in the early fifth instar, then INS-a mRNA declined prematurely whereas INS-b mRNA remained similar to that in the intact larvae. By contrast, in the fat body of -CA larvae, the exogenous JH had little effect on the levels of INS-a mRNA, but enhanced expression of INS-b mRNA relative to intact larvae. Culture of epidermis from day 1 fifth instar larvae with 40 ng/ml 20E for up to 24 h accelerated the loss of INS-a mRNA without affecting the levels of INS-b mRNA. Both mRNAs declined in isolated larval abdomens over a 24 h period, and this decline was slowed by 1 µg methoprene (a JH analog). Together these results indicate that JH controls the levels of the two mRNAs in both the epidermis and fat body, with additional factors involved in regulating these genes in the fat body during the molt and in the epidermis during the growth phase.