Up-regulation of HO-1 expression induced by LXA₄ in cardiomyocytes.

<p>In A, cardiomyocytes were pretreated with LXA₄ (10 nM) for 1,6,12 and 24 h before H/R exposure. In B and C, cardiomyocytes were preincubated with LXA₄ (0, 0.1, 1, 10 nM) for 12 h before H/R exposure. In A and B, the expressions of HO-1 mRNA and protein were measured by using real-time RT-PCR and Western blot respectively. The amount of PCR products was normalized with β-actin to determine the relative expression ratio (log2 fold) for each mRNA. The Western blotting of tubulin protein in lower panel served as a loading control. Protein expression of HO-1 was shown as HO-1/tubulin ratio for each sample. In C, HO-1 activity was expressed as picomoles of bilirubin formed per milligram of protein per hour. The HO-1 levels in cell lysates were assessed by ELISA. In D, the expression and the localization of HO-1 in the cells treated with or without H/R injury and with or without LXA₄ pretreatment were investigated by using fluorescence microscopy (×400). Values were and mean ± SEM of three independent experiments. *<i>P</i><0.05, as compared to the cells exposed to H/R injury alone.</p

LXA₄-stimulated HO-1 expression is dependent on formation of Nrf2/ARE complex.

<p>In A, cardiomyocytes were transfected with mouse HO-1 promoter construct (E1) or a mutated mouse HO-1 promoter construct (M739) with or without transfection of dominant-negative Nrf2 (dnNrf2), and then treated with H/R injury with or without pretreatment with LXA₄. Fold induction of luciferase activity of HO-1 promoter was assessed by employing the reporter gene transfection assays. In B, nuclear extracts were prepared from cardiomyocytes, and then subjected to EMSA with biotin-labeled double-stranded oligonucleotide probe of ARE. Supershift assay was performed using an antibody against to Nrf2. In C, ChIP assay determining Nrf2 binding to the HO-1 E1 enhancer following treatment of the cells with H/R injury, LXA₄ or H/R injury plus LXA₄. Results are representative of three independent experiments. In A, *<i>P</i><0.05, as compared to the cells without treatment.</p

Cellular morphology, viability, LDH and CK release.

<p>Figure A to H showed the morphology of rat cardiomyocyte monolayer (×400). Control cells were not treated (A). In B, F, G and H, the cells were exposed to H/R injury for 12 h. In C and D, the cells were treated with LXA₄ or ZnPP-IX respectively, without H/R injury. The arrows denote the necrotic cells with pyknotic nuclei. In F, cardiomyocytes was pretreated with LXA₄ for 12 h and then exposed to H/R injury. In E and H, cardiomyocytes were preincubated with LXA₄ (10 nM) 12 h, and coincubation was continued for 12 h with ZnPP-IX (10 µM) with or without H/R injury respectively. In I, cardiomyocyte viability was as measured by MTT assay. The cells without any treatment (A) served as controls. The letters A to H within the columns represent the same treatment of the cells as shown in Figure A to H. In J, the levels of LDH and CK in the cell supernatants were shown respectively. Results are representative of five independent experiments. Values are means ± SEM, *<i>P</i><0.05, as compared to the control cells without treatment.</p

Synthesis of Pyrrolo(spiro-[2.3′]-oxindole)-spiro-[4.3″]-oxindole via 1,3-Dipolar Cycloaddition of Azomethine Ylides with 3‑Acetonylideneoxindole

A series of novel dispirooxindole derivatives, 3-acetyl-5-phenyl-pyrrolo­(spiro-[2.3′]-1′-benzyl-oxindole)-spiro-[4.3″]-1″-benzyl-oxindoles, were synthesized via 1,3-dipolar cycloaddition of the azomethine ylide with 3-acetonylideneoxindole in high regioselectivities and yields. An unusual regioselectivity was observed in this 1,3-dipolar cycloaddition, leading to the construction of novel dispirooxindole skeleton. The substituent effects on the regioselectivity were also investigated

Synthesis of Pyrrolo(spiro-[2.3′]-oxindole)-spiro-[4.3″]-oxindole via 1,3-Dipolar Cycloaddition of Azomethine Ylides with 3‑Acetonylideneoxindole

A series of novel dispirooxindole derivatives, 3-acetyl-5-phenyl-pyrrolo­(spiro-[2.3′]-1′-benzyl-oxindole)-spiro-[4.3″]-1″-benzyl-oxindoles, were synthesized via 1,3-dipolar cycloaddition of the azomethine ylide with 3-acetonylideneoxindole in high regioselectivities and yields. An unusual regioselectivity was observed in this 1,3-dipolar cycloaddition, leading to the construction of novel dispirooxindole skeleton. The substituent effects on the regioselectivity were also investigated

Synthesis of Pyrrolo(spiro-[2.3′]-oxindole)-spiro-[4.3″]-oxindole via 1,3-Dipolar Cycloaddition of Azomethine Ylides with 3‑Acetonylideneoxindole

A series of novel dispirooxindole derivatives, 3-acetyl-5-phenyl-pyrrolo­(spiro-[2.3′]-1′-benzyl-oxindole)-spiro-[4.3″]-1″-benzyl-oxindoles, were synthesized via 1,3-dipolar cycloaddition of the azomethine ylide with 3-acetonylideneoxindole in high regioselectivities and yields. An unusual regioselectivity was observed in this 1,3-dipolar cycloaddition, leading to the construction of novel dispirooxindole skeleton. The substituent effects on the regioselectivity were also investigated

<i>L</i>‑Pyroglutamic Sulphonamide as Hydrogen-Bonding Organocatalyst: Enantioselective Diels–Alder Cyclization to Construct Carbazolespirooxindoles

Hydrogen-bonding organocatalysts <i>L</i>-pyroglutamic sulphonamides were readily synthesized for the first time by fully exploiting the potentials of <i>L</i>-pyroglutamic acid. The newly designed catalyst was successfully applied in catalyzing asymmetric Diels–Alder cyclization of methyleneindolinones with 2-vinyl-1<i>H</i>-indoles to efficiently assemble carbazolespirooxindoles in excellent stereoselectivity (up to 99% ee, >20:1 dr) and yields (up to 99%). Mechanistic studies disclosed that the well-designed hydrogen-bonding modes between <i>L</i>-pyroglutamic sulphonamide and substrates were crucial for stereocontrol in the cyclization

Organocatalytic Asymmetric Allylic Alkylation of Morita–Baylis–Hillman Carbonates with Diethyl 2‑Aminomalonate Assisted by In Situ Protection

With the aid of in situ protection by <i>N</i>-(2-formylphenyl)-4-methyl-benzenesulfonamide, enantioselective allylic alkylation of Morita–Baylis–Hillman carbonates with diethyl 2-aminomalonate was successfully realized. The corresponding adducts can be obtained in up to 99% yield with up to 98% ee as well as excellent regioselectivity. Besides, the adducts with opposite configurations were readily prepared by utilizing easily available and inexpensive quinine or quinidine as organocatalyst. Facile deprotection of the resulting adduct provides straightforward access to enantiopure α-methylene-γ-lactam

A new flavanone from the aerial parts of <i>Penthorum chinense</i>

<div><p>A new flavanone, 5-methoxy-pinocembrin-7-<i>O-</i>β-d-glucoside (<b>1</b>), together with six known compounds, quercetin-3-<i>O-</i>β-d-xyloside (<b>2</b>), quercetin-3′-<i>O-</i>α-l-rhamnoside (<b>3</b>), quercetin (<b>4</b>), pinocembrin-7-<i>O-</i>β-d-glucoside (<b>5</b>), pinocembrin (<b>6</b>), gallic acid (<b>7</b>), were isolated and identified from the aerial parts of <i>Penthorum chinense</i>. Compounds <b>2</b> and <b>3</b> were isolated from this plant for the first time.</p></div

Additional file 3: Table S1. of Profiling of the perturbed metabolomic state of mouse spleen during acute and chronic toxoplasmosis

Summary results of the differential ions. (DOC 32 kb