[Determination of short- and medium-chain chlorinated paraffins in different components of human blood using gas chromatography-electron capture negative ion-low resolution mass spectrometry].

Short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) have attracted significant attention because of their persistence, biotoxicity, bioaccumulation, and long-range migration. Given their worldwide detection in a variety of environmental matrices, concerns related to the high exposure risks of SCCPs and MCCPs to humans have grown. Thus, knowledge of the contamination patterns of SCCPs and MCCPs and their distribution characteristics in the vivo exposure of humans is of great importance. However, little information is available on the contamination of SCCPs and MCCPs in human blood/plasma/serum, mainly because of the difficulty of sample preparation and quantitative analysis. In this study, a new blood sample pretreatment method based on Percoll discontinuous density gradient centrifugation was developed to separate plasma, red blood cells, white blood cells, and platelets from human whole blood. A series of Percoll sodium chloride buffer solutions with mass concentrations of 1.095, 1.077, and 1.060 g/mL were placed in a centrifuge tube from top to bottom to establish discontinuous density gradients. The dosage for each density gradient was 1.5 mL. Human whole blood samples mixed with 0.85% sodium chloride aqueous solution were then added to the top layer of the Percoll sodium chloride solution. After centrifugation, the whole blood was separated into four components. The plasma was located at the top layer of the centrifuge tube, whereas the platelets, white blood cells, and red blood cells were retained at the junction of the various Percoll sodium chloride solutions. The sampling volume of human whole blood and incubation time were optimized, and results indicated that an excessively long incubation time could lead to hemolysis, resulting in a decrease in the recoveries of SCCPs and MCCPs. Therefore, a sampling volume of 1.5 mL and incubation time of 10 min at 4 ℃ were adopted. The cells of the blood components were further broken and extracted by ultrasonic pretreatment, followed by multilayer silica gel column chromatography for lipid removal. The use of 80 mL of n-hexane-dichloromethane (1∶1, v/v) and 50 mL of dichloromethane as the elution solvents (collected together) for the gel column separated the SCCPs and MCCPs from the lipid molecules in the blood samples. Gas chromatography-electron capture negative ion-low resolution mass spectrometry (GC-ECNI-LRMS) was used to determine the SCCPs and MCCPs. Quantification using the corrected total response factor with degrees of chlorination was achieved with linear corrections (R2=0.912 and 0.929 for the SCCPs and MCCPs, respectively). The method detection limits (MDLs) for the SCCPs and MCCPs were 1.57 and 8.29 ng/g wet weight (ww, n=7), respectively. The extraction internal standard recoveries were 67.0%-126.6% for the SCCPs and 69.5%-120.5% for the MCCPs. The developed method was applied to determine SCCPs and MCCPs in actual human whole blood samples. The contents of SCCPs and MCCPs were 10.81-65.23 and 31.82-105.65 ng/g (ww), respectively. Red blood cells exhibited the highest contents of CPs, followed by plasma, white blood cells, and platelets. The proportions of SCCPs and MCCPs in red blood cells and plasma were 70% and 66%, respectively. In all four components, the MCCP contents were higher than the SCCP contents, and the ratios of MCCPs to SCCPs ranged from 1.04 to 3.78. Similar congener patterns of SCCPs and MCCPs were found in the four components of human whole blood. C10-CPs and C14-CPs were predominantly observed in the SCCPs and MCCPs, respectively. In summary, a simple and efficient method was proposed to determine low concentrations of SCCPs and MCCPs in human blood with high sensitivity and selectivity. This method can meet requirements for the quantitative analysis of SCCPs and MCCPs in human blood components, thereby providing technical support for human health risk assessment.

Potential suppressive effects of theophylline on human rectal cancer SW480 cells in vitro by inhibiting YKL-40 expression.

Chitinase-3-like-1 protein (YKL-40), a member of the mammalian chitinase-like glycoproteins, serves a key role in the pathogenesis of rectal cancer. The present study examined the antitumor effect of theophylline, a pan-chitinase inhibitor, in rectal cancer in vitro and investigated the mechanism by which it acted. SW480 cell lines were treated with varying theophylline concentrations (10-2, 10-3, 10-4 and 10-5 mol/l). An MTT assay was used to observe cell proliferation and identify the optimal theophylline concentration. Western blotting was used to analyze YKL-40 expression. The cell cycle distribution of SW480 cell lines treated with theophylline was measured by flow cytometry. The angiopoietin-2 expression level was measured by ELISA. The expression levels of YKL-40 were evidently decreased in theophylline-treated SW480 cell lines. The proliferation of SW480 cells was inhibited following theophylline treatment, which was associated with G1 phase cell cycle arrest and a decrease in the expression of angiopoietin-2. The mechanism of theophylline action may involve the downregulation of YKL-40 expression, arrest of the cell cycle at G1 phase and inhibition of angiopoietin-2 expression. These results provide a rationale for the potential use of anti-YKL-40 and anti-angiogenic strategies in treating rectal cancer.

Effect of calcium or vitamin D supplementation on vascular outcomes: a meta-analysis of randomized controlled trials.

BACKGROUND: Whether calcium or vitamin D supplementation reduces serious vascular outcomes in older people remains unclear. We conducted a meta-analysis based on randomized controlled trials to evaluate the effect of calcium or vitamin D supplementation on the risk of major cardiovascular outcomes. METHODS: We performed electronic searches in PubMed, Embase, and the Cochrane Library to identify relevant randomized controlled trials. Odds ratios (ORs) were used to measure the effect of calcium or vitamin D supplementation on the risk of major vascular outcomes with a random-effect model. RESULTS: Of the 1643 identified studies, we included 11 trials reporting data on 50,252 individuals. These studies reported 2685 major cardiovascular events, 1097 events of myocardial infarction, and 1350 events of stroke. Calcium or vitamin D supplementation did not have an effect on major cardiovascular events (OR, 1.03; 95% confidence interval [CI]: 0.94-1.12; P=0.54), myocardial infarction (OR, 1.08; 95% CI: 0.96-1.22; P=0.21), or stroke (OR, 1.01; 95% CI: 0.91-1.13; P=0.80) when compared to the effect with a placebo. Subgroup analysis indicated that calcium supplementation alone might play an important role in increasing the risk of major cardiovascular events, myocardial infarction, and stroke, but this difference could not be identified as statistically significant. Furthermore, males seem to experience more harmful effects with supplements of calcium or vitamin D than the effects experienced by females. CONCLUSIONS: Calcium supplementation might increase the risk of major cardiovascular events, myocardial infarction, and stroke compared to the risk with a placebo.

Using Ruditapes philippinarum conglutination mud to produce bioflocculant and its applications in wastewater treatment.

A novel bioflocculant-producing bacterium, ZHT4-13, was isolated from Ruditapes philippinarum conglutination mud. By biomicroscope morphological observation, 16S rDNA sequence identification and physiological and biochemical characteristics, strain ZHT4-13 was identified as Rothia sp. The bioflocculant MBF4-13 produced by strain ZHT4-13 had a flocculating efficiency of 86.22% for 5 g L(-1) Kaolin clay suspension when the initial pH was 9 and the temperature was 20 degrees C. It had flocculating effect in a wide range, pH 1-13 and temperature 4-100 degrees C. Analysis of MBF4-13 by UV-Vis spectrophotometer, Fourier-transform infrared spectrophotometer (FT-IR) and (1)H nuclear magnetic resonance (NMR) indicated that the main component of MBF4-13 is polysaccharide. The culture conditions to produce strain ZHT4-13 were optimized with orthogonal design of experiments. MBF4-13 had high efficiency in decolorizing dye solutions, had some abilities to remove heavy metal ions (Cr(2)O(7)(2-), Ni(2+)) and improve performance of activated sludge.