Regulation of anti-apoptotic and pro-apoptotic protein expression caused by tamoxifen are not affected by risperidone.

<p>Cells were treated with 1 µM tamoxifen with or without 3 µM risperidone or 0.3 µM fluoxetine for 72 hours. Treatment of tamoxifen with or without risperidone resulted in decreased protein expression of Bcl-2 (A) and Bcl-x_L (B). Protein expression of Bax (C) and Bak (D) were increased by tamoxifen with or without risperidone. Graphs show mean ± SEM of three or more independent experiments. *, <i>p</i><0.05 to control group; #, <i>p</i><0.05 to tamoxifen-treated group; t-test. Tam, tamoxifen; Ris, risperidone; Flx, fluoxetine.</p

Decrease of tumor volume by tamoxifen is not influenced by combination treatment of risperidone.

<p>In T47D-inoculated female nude mice, tumor masses had been recorded since day 21, and the individual or combination regimens of each drug [25 µg tamoxifen (Tam) per mouse or/and 2.5 µg risperidone (Ris) per mouse] were started at Day42 with intraperitoneal injection every 2 days. Tumor volume (tumor volume  =  length×width²×0.5) was recorded once a week until Day 91. Graph shows mean ± SEM of four mice in each group.</p

T47D human breast cancer cells exhibit tamoxifen-induced cytotoxic effect dose- and time-dependently.

<p>(<b>A</b>) T47D human breast cancer cells but not MCF-7 cells expressed significant amount of CYP2D6 protein. A549 human lung cancer was loaded as positive control which shows prominent protein expression. (<b>B</b>) Cells were treated with tamoxifen, 4-OH-tamoxifen, and endoxifen (0.1–3 µM) for 7 days. Cell viability of T47D cells was examined by crystal violet (CV) staining (<b>C, D</b>) Tamoxifen (1 µM), 4-OH-tamoxifen (1 µM), and endoxifen (1 µM) markedly inhibited cell viability from Day3 to Day7 in T47D cells, measured by both crystal violet (CV) staining and MTT assay. Graphs show mean ± S.E.M. of at least three independent experiments. <b>*</b>, <i>p</i><0.05 to control group; t-test. Tam, tamoxifen; 4-OHTam, 4-hydroxy-tamoxifen; Endx, endoxifen.</p

Tamoxifen-induced cell cycle arrest in G0/G1 phase is not interfered by risperidone in T47D cells.

<p>Cells were treated with tamoxifen (1 µM), 4-OH-tamoxifen (1 µM), endoxifen (1 µM), risperidone (3 µM), paliperidone (3 µM), tamoxifen with 3 µM risperidone, or tamoxifen with 0.3 µM fluoxetine for 7 days. Cell viability was examined by crystal violet staining (<b>B</b>), MTT assay (<b>C</b>), and SRB assay (<b>D</b>). Representative crystal violet staining was shown as (<b>A</b>) and quantified by spectrophotometry (<b>B</b>). (<b>E</b>, <b>F</b>) Tamoxifen-induced cytostasis analyzed by flow cytometry showed that cell cycle was arrested at G0/G1 phase, since the percentage of cells at G0/G1 phase was markedly increased, and percentage of cells at S and G2/M phase were decreased respectively by tamoxifen treatment for 2 days. Graphs show mean ± S.E.M. of at least three independent experiments. <b>*</b>, <i>p</i><0.05 to control group; #, <i>p</i><0.05 to tamoxifen-treated group; t-test. Tam, tamoxifen; 4-OHTam, 4-hydroxy-tamoxifen; Endx, endoxifen; Ris, risperidone; Pali, paliperidone; Flx, fluoxetine.</p

Risperidone shows no interference in tamoxifen-induced cytotoxic effect in T47D cells.

<p>Cells were treated with risperidone (<b>A</b>) or its main metabolite paliperidone (0.01–10 µM) (<b>B</b>) for 7 days. Cells were treated with control vehicle (closed bar) or 1 µM tamoxifen (open bar) with or without risperidone (<b>C</b>) or fluoxetine (<b>D</b>) for 7 days. Cell viability of T47D cells was examined by crystal violet (CV) staining. Graphs show mean ± S.E.M. of at least three independent experiments. <b>*</b>, <i>p</i><0.05 to control vehicle group; #, <i>p</i><0.05 to control tamoxifen alone group; t-test.</p

Risperidone has no influence on tamoxifen-induced cleavage of caspases and PARP-1 in T47D cells.

<p>Cells were treated with 1 µM tamoxifen with or without 3 µM risperidone or 0.3 µM fluoxetine for 72 hours. Representative protein blotting images are shown in (<b>A</b>). Treatment of tamoxifen with or without risperidone resulted in increased protein expression of cleaved caspase 9 (<b>B</b>), caspase 7 (<b>C</b>), caspase 3 (<b>D</b>), and PARP-1 (<b>E</b>). Graphs show mean ± SEM of three or more independent experiments. <b>*</b>, <i>p</i><0.05 to control group; #, <i>p</i><0.05 to tamoxifen-treated group; t-test. Tam, tamoxifen; Ris, risperidone; Flx, fluoxetine.</p

Tamoxifen down-regulates cell cycle regulators pRb, cyclin D1 and oncoprotein c-Myc without disturbing by risperidone.

<p>Cells were treated with 1 µM tamoxifen with or without 3 µM risperidone or 0.3 µM fluoxetine for 48 hours. Protein expression of cell cycle regulators pRb (A) and cyclin D1 (B) and oncoprotein c-Myc (C) were measured by Western blotting. Graphs show mean ± SEM of three or more independent experiments. *, <i>p</i><0.05 to control group; #, <i>p</i><0.05 to tamoxifen-treated group; t-test. Tam, tamoxifen; 4-OHTam, 4-hydroxy-tamoxifen; Endx, endoxifen; Ris, risperidone; Pali, paliperidone; Flx, fluoxetine.</p

Associations between the Duration of Dialysis, Endotoxemia, Monocyte Chemoattractant Protein-1, and the Effects of a Short-Dwell Exchange in Patients Requiring Continuous Ambulatory Peritoneal Dialysis

<div><p>Background</p><p>Endotoxemia is exaggerated and contributes to systemic inflammation and atherosclerosis in patients requiring continuous ambulatory peritoneal dialysis (CAPD). The risk of mortality is substantially increased in patients requiring CAPD for >2 years. However, little is known about the effects of long-term CAPD on circulating endotoxin and cytokine levels. Therefore, the present study evaluated the associations between plasma endotoxin levels, cytokine levels, and clinical parameters with the effects of a short-dwell exchange on endotoxemia and cytokine levels in patients on long-term CAPD.</p><p>Methods</p><p>A total of 26 patients were enrolled and divided into two groups (short-term or long-term CAPD) according to the 2-year duration of CAPD. Plasma endotoxin and cytokine levels were measured before and after a short-dwell exchange (4-h dwell) during a peritoneal equilibration test (a standardized method to evaluate the solute transport function of peritoneal membrane). These data were analyzed to determine the relationship of circulating endotoxemia, cytokines and clinical characteristics between the two groups.</p><p>Results</p><p>Plasma endotoxin and monocyte chemotactic protein-1 (MCP-1) levels were significantly elevated in the long-term group. PD duration was significantly correlated with plasma endotoxin (<i>r</i> = 0.479, <i>P</i> = 0.016) and MCP-1 (<i>r</i> = 0.486, <i>P</i> = 0.012). PD duration was also independently associated with plasma MCP-1 levels in multivariate regression. Plasma MCP-1 levels tended to decrease (13.3% reduction, <i>P</i> = 0.077) though endotoxin levels did not decrease in the long-term PD group after the 4-h short-dwell exchange.</p><p>Conclusion</p><p>Long-term PD may result in exaggerated endotoxemia and elevated plasma MCP-1 levels. The duration of PD was significantly correlated with circulating endotoxin and MCP-1 levels, and was an independent predictor of plasma MCP-1 levels. Short-dwell exchange seemed to have favorable effects on circulating MCP-1 levels in patients on long-term PD.</p></div

Comparison of circulating endotoxin and cytokine levels after a 4-h exchange between the short- and long-term PD groups.

<p>Values are median (interquartile range) or means±standard error of the mean.</p><p>% change, percentage change; GM-CSF, granulocyte-macrophage colony-stimulating factor; IFN-γ, interferon-γ; IL, interleukin; IL-1RA, interleukin-1 receptor antagonist; MCP-1, monocyte chemoattractant protein-1; TNF, tumor necrosis factor.</p><p>Comparison of circulating endotoxin and cytokine levels after a 4-h exchange between the short- and long-term PD groups.</p

Correlations between PD duration, clinical parameters, and cytokine levels.

<p>CCI, Charlson's comorbidity index; iPTH, intact parathyroid hormone; MCP-1 monocyte chemoattractant protein-1; PD, peritoneal dialysis; <i>r</i>, Spearman's rank correlation coefficient.</p><p>Correlations between PD duration, clinical parameters, and cytokine levels.</p