OEOA was retained for a longer period in mouse blood plasma than OA.

<p>MS-MS product ion mass spectra of OEOA (A) and OA (B). (C) K562 cells were treated with OA or OEOA (1 µM) for 6 h. Culture media and cell lysates were collected at indicated time points for HPLC-MS/MS analysis. Data was represented as mean ± SEM in cell lysate compared to total exposure, n = 2 (* <i>p</i><0.05). (D) Concentrations of OA and OEOA in mouse plasma after intraperitoneal injection. Blood was collected from mice at different time points after single administration of OA and OEOA. Data was represented as mean ± SEM, n = 2 animals per time point (* <i>p</i><0.05).</p

OEOA attenuated phosphorylation of Rb protein in K562 and Jurket cells.

<p>(A) K562 and Jurket cells were treated with OEOA (0.1–10 µM) for 2 days, and the cell lysates were subjected to Western blot analysis for p-Rb and Rb. Actin served as an equal loading control. Histograms in (B) show the relative expression of p-Rb (normalized to actin) as compared to the vehicle-treated cells. Results were representative blots from three separate experiments, (* <i>p</i><0.05).</p

OEOA promoted erythroid differentiation in K562 cells.

<p>K562 cells were treated with OEOA (0.1–10 µM) for 2 days. Total RNA was reverse transcribed and subjected to real time-PCR analysis with primers specific to <i>γ-globin</i> (A) and <i>cd41b</i> (B), respectively. <i>hprt1</i> served as an internal housekeeping gene control. Data were expressed as fold change to the control cells as mean ± SEM of three independent experiments (* <i>p</i><0.05). (C) K562 cells were treated with OEOA (0.1–10 µM) for 2 days. Western blot analysis of Bcr-Abl and Erk1/2 was performed. Actin served as an equal loading control. Histograms on the right show the relative expression of various proteins (normalized to actin) as compared to the control cells. Results were representative blots from three separate experiments, (* <i>p</i><0.05).</p

OEOA inhibited cell proliferation in leukemia cell lines.

<p>Different cell lines were treated with various concentrations of OEOA or OA for 2 days. Cell growth was measured by MTT assay: (A) K562, (B) HEL, (C) Jurket (D) HEKneo, and (E) HepG2, MCF<b>-</b>7 and HeLa cells. Data are mean ± SEM of three independent experiments (* <i>p</i><0.05).</p

OEOA did not induce cell death in K562 and HEL cells.

<p>K562 (A & C) and HEL (B & D) cells were treated with OEOA (1 µM) for 6 days. Cell viability was measured by trypan blue exclusion as described in Materials and Methods. Data are mean ± SEM of three independent experiments (* <i>p</i><0.05).</p

OEOA induced G1 cell cycle arrest in K562 cells.

<p>(A) Cells were incubated with OEOA (1 or 10 µM) for 24 h. The distribution of cell cycle was examined by PI staining method. The table summarized the distribution of cells in OEOA-treated or control cells. Data represented mean ± SEM of three independent experiments (* <i>p</i><0.05). (B) K562 cells were cultured in the presence of OEOA (1 or 10 µM) for 2 days. Total proteins were collected for Western blot analysis to detect the expression of p27, Cdk4, Cdk6, Cyclin D1, Cyclin E and RAMP. Actin served as an equal loading control. Histograms on the right show the relative expression of various proteins (normalized to actin) as compared to the control cells. Results were representative blots from three separate experiments, (* <i>p</i><0.05).</p

Diarylheptanoids from Rhizomes of Alpinia officinarum Inhibit Aggregation of α‑Synuclein

Two new diarylheptanoids, alpinin A (<b>1</b>) and alpinin B (<b>2</b>), together with 18 known diarylheptanoids (<b>3</b>–<b>20</b>), were isolated from the rhizomes of Alpinia officinarum. Their structures were elucidated by comprehensive spectroscopic analysis, including high-resolution mass spectrometry, infrared spectroscopy, and one- and two-dimensional nuclear magnetic resonance spectroscopy. Structurally, alpinin A is a new member of the small family of oxa-bridged diarylheptanoids and contains the characteristic 2,6-<i>cis</i>-configured tetrahydropyran motif (C₁–C₅ oxa bridge). The absolute configuration of alpinin A was confirmed by asymmetric total synthesis of the enantiomer (<i>ent</i>-<b>1</b>), corroborating the assignment of the molecular structure. The absolute configuration of alpinin B was determined on the basis of the analysis of the circular dichroism exciton chirality spectrum. We evaluated the inhibitory activity of all isolated diarylheptanoids against α-synuclein aggregation at 10 μM. Alpinins A and B significantly inhibited α-synuclein aggregation by 66 and 67%, respectively

Two pairs of unusual melibiose and raffinose esters from <i>Scrophularia ningpoensis</i>

<p>A pair of unusual melibiose esters (<b>1<i>α</i></b>/<b>1<i>β</i></b>) and a pair of unusual raffinose esters (<b>2<i>α</i>/2<i>β</i></b>), were isolated from <i>Scrophularia ningpoensis</i>. Structures of them were established by detailed spectroscopic analyses to be 6-<i>O</i>-(<i>E</i>)-cinnamoyl-<i>α</i>-d-galactopyranosyl-(1→6)-<i>α</i>(<i>β</i>)-d-glucopyranose (<b>1<i>α</i></b>/<b>1<i>β</i></b>) and 6-<i>O</i>-(<i>E</i>)/(<i>Z</i>)-cinnamoyl-<i>α</i>-d-galactopyranosyl-(1→6)-<i>α</i>-d-glucopyranosyl-(1→2)-<i>β</i>-d-fructofuranose (<b>2<i>α</i>/2<i>β</i></b>), respectively. All these compounds were evaluated for antifouling activity against the settlement of <i>Balanus amphitrite</i> larvae, along with the cytotoxic effect against the proliferation of HeLa cell lines.</p

AA3 reduces the disease severity of EAE mice.

<p>(A) We orally administered AA3 or water (control) daily starting from the day of immunization. The time courses of symptom development (reflected by the EAE score) and the incidence of EAE scores at day 20 are shown (table below graph). (B) We orally administered AA3 or water (control) to EAE mice daily starting from day 8 post-immunization; the time courses of EAE scores are shown. Data are mean ± SEM (<i>n</i> = 24 per treatment group); *<i>p</i> < 0.05, Student’s <i>t</i>-test. (C) Histopathologic examination of spinal cord tissues by hematoxylin and eosin (top and middle panels) and Luxol Fast Blue staining (bottom panels). We obtained tissues from mice injected with complete Freund’s adjuvant (CFA), or EAE mice treated with water (Con/EAE) or AA3 (AA3/EAE). Representative pictures are shown. Scale bar, 100 μm.</p