Cleavage of AUF1 by Coxsackievirus B Affects DDX5 Regulatory on Viral Replication through iTRAQ Proteomics Analysis.

Coxsackievirus B (CVB) 3C protease (3Cpro) plays a specific cleavage role on AU-rich binding factor (AUF1, also called hnRNP D), which consequently disputes the regulation of AUF1 on downstream molecules. In our study, the iTRAQ approach was first used to quantify the differentially expressed cellular proteins in AUF1-overexpressing HeLa cells, which provides straightforward insight into the role of AUF1 during viral infection. A total of 1,290 differentially expressed proteins (DEPs), including 882 upregulated and 408 downregulated proteins, were identified. The DEPs are involved in a variety of cellular processes via GO terms, protein-protein interactions, and a series of further bioinformatics analyses. Among the DEPs, some demonstrated important roles in cellular metabolism. In particular, DDX5 was further verified to be negatively regulated by AUF1 and increased in CVB-infected cells, which in turn promoted CVB replication. These findings provide potential novel ideas for exploring new antiviral therapy targets.

Comparison of intravenous and oral acetaminophen for pain control after total knee and hip arthroplasty: A systematic review and meta-analysis.

OBJECTIVE: To evaluate the efficacy between intravenous and oral acetaminophen as adjunct to multimodal analgesia regimens for pain management after total knee and hip arthroplasties. METHODS: We conduct electronic searches of Medline (1966-2017.09), PubMed (1966-2017.09), Embase (1980-2017.09), ScienceDirect (1985-2017.09), and the Cochrane Library. Only randomized controlled trials (RCTs) are included. The quality assessment is performed according to the Cochrane systematic review method. Fixed/random effect model is adopted according to the heterogeneity tested by I statistic. Meta-analysis is performed using Stata 11.0 software. RESULTS: Two RCTs are included involving 236 patients. The present meta-analysis demonstrated that there were no significant differences between groups regarding pain scores at 12, 24, or 48 hours. No significant differences were observed in terms of opioid consumption at 12, 24, or 48 hours after arthroplasties. CONCLUSION: Intravenous acetaminophen to multimodal analgesia dose not demonstrate a significant benefit in reducing pain and opioid consumption compared oral formulation after total knee arthroplasty and total hip arthroplasty. Higher-quality RCTs are required for further research.

miR-330 regulates interleukin-13-induced MUC5AC secretion by targeting Munc18b in human bronchial epithelial cells.

Mucus hypersecretion by airway epithelium and plugging of the airways are primary reasons of mortality in asthma patients and major causes of asthma disease progression and exacerbation. MUC5AC protein is a major component of airway mucus. MicroRNAs (miRNAs), a class of small noncoding RNAs, have emerged as moderators of MUC5AC production and secretion and are implicated in the pathogenesis of asthma. Recently, miR-330 has been reported to be downregulated in the blood of asthmatic patients, acting as a biomarker for asthma. The role of miR-330 in asthma, however, is unclear. Here, we showed that interleukin (IL)-13 induced MUC5AC secretion and inhibited miR-330 expression in a concentration-dependent manner in human bronchial epithelial cells (HBE16). Upregulation of miR-330 in HBE16 cells inhibited IL-13-induced MUC5AC secretion while, conversely, depletion of endogenous miR-330 exacerbated MUC5AC secretion. Munc18b (Syntaxin-Binding Protein 2; STXBP2) is a limiting component of the exocytic machinery of airway epithelial cells. We identified and validated that Munc18b was a direct target of miR-330 and miR-330 regulated MUC5AC secretion in HBE16 cells by acting directly on the 3'UTR of Munc18b mRNA. Collectively, these data reveal that miR-330 inhibits IL-13-induced MUC5AC secretion in human bronchial epithelial cells by targeting Munc18b, encouraging us to further explore the potential of manipulating miR-330 in treatment of airway diseases with mucus hypersecretion.

[Detection method of phthalate esters and their metabolites in blood].

OBJECTIVE: A method for determination of phthalate esters and their metabolites in blood by using SPE-ultra performance liquid chromatography tandem mass spectrometry. METHODS: A stable isotope detection method been developed for the determination of 20 kinds of phthalate esters and 6 kinds of phthalate esters metabolites in blood by ultra-performance liquid chromatography-mass spectrometry( UPLC-MS) coupled with SPE column purification. Blood samples were diluted with acetate buffer( pH5. 2), enzymatic hydrolyzed by ß-glucuronidase for 12 h, then purified with HLB column. An ACQUITYBEH Phenyl column( 2. 1 mm ×100 mm, 1. 7 µm) was used for separation by the gradient elution with acetonitrile and aqueous solution containing 0. 02% formic acid as the mobile phases. In the present study, an electrospray ionization( ESI) source was used, and the internal standard method was used for quantitation. RESULTS: The linear ranges of the 26 analytes were from 1. 0- 20. 0 µg/L, the coefficients of correlation were greater than 0. 995. The limits of detection( LODs) of the 26 analytes were all lower than1. 0 µg/L. Recoveries studies were carried out using serum samples fortified with the 26 analytes at the levels of 2. 5, 5. 0 and 10. 0 µg/L, recoveries were obtained in the range of63. 0%- 97. 7% with relative standard deviations( RSDs) from 1. 9% to 19. 1%. CONCLUSION: The established method is accurate and highly sensitive, and can be used for the qualitative and quantitative analysis of the residues of the phthalate esters and their metabolites in the blood samples.

Novel dispersive micro-solid-phase extraction combined with ultrahigh-performance liquid chromatography-high-resolution mass spectrometry to determine morpholine residues in citrus and apples.

This paper presents a new analytical method for the determination of morpholine residues in citrus and apples using a novel dispersive micro-solid-phase extraction (DMSPE), followed by ultrahigh-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS). Samples were extracted with 1% formic acid in acetonitrile/water (1:1, v/v) and then cleaned up using the DMSPE procedure. Morpholine from the extract was adsorbed to a polymer cation exchange sorbent and eluted with ammonium hydroxide/acetonitrile (3:97, v/v) through a 1 mL syringe with a 0.22 µm nylon syringe filter. All of the samples were analyzed by UHPLC-HRMS/MS on a Waters Acquity BEH hydrophilic interaction chromatography column using 0.1% formic acid and 4 mM ammonium formate in water/acetonitrile as the mobile phase with gradient elution. The method showed good linearity (R(2) > 0.999) in the range of 1-100 µg/L for the analyte. The limit of detection and limit of quantitation values of morpholine were 2 and 5 µg/kg, respectively. The average recoveries of morpholine from the citrus and apple samples spiked at three different concentrations (5, 20, and 100 µg/kg) were in a range from 78.4 to 102.7%.

[Determination of 18 pesticide residues in red wine by ultra high performance liquid chromatography-high resolution mass spectrometry with isotope dilution technique].

A method for the simultaneous determination of 18 pesticide residues in red wine was developed using ultra high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) with isotope dilution technique. The red wine samples were extracted with acetonitrile, and the extracts were cleaned up with dispersive solid phase extraction (dSPE) using the mixture of N-propyl ethylene diamine (PSA) and C18 powder as sorbent. The extracted components were separated on a BEH C18 column by gradient elution. The qualitative and quantitative analyses were operated under full scan/data dependent MS/MS (ddms2) and targeted selective ion monitoring (tSIM) by high resolution mass spectrometry, respectively. Carbendazim-D4, chlorpyrifos-D10, imidacloprid-D4, methoxyfenozide-D9, pyrimethanil-D5 and tebuconazole-D6 were used as the internal standards to reduce the matrix effects. The response of each pesticide showed a good linearity in the range of 0.5-50 microg/kg with the correlation coefficient more than 0.999. The limits of detection and quantification for the 18 pesticides in the spiked blank red wine were 0.5 microg/kg and 1.0 microg/kg, respectively. The recovery results with spiked blank red wine samples at the levels of 1 to 40 microg/kg were satisfactory with average recoveries of 85.4% - 117.9% and the RSDs of 0.5%-6.1%. The method was applied for the determination of the red wine real samples from the market. Carbendazim, imidacloprid, pyrimethanil, tebuconazole and triadimenol were detected in the samples. The results show that the method is suitable for the rapid screening and quantitative analysis of pesticide residues in red wine.

Organochlorine pesticides and pyrethroids in Chinese tea by screening and confirmatory detection using GC-NCI-MS and GC-MS/MS.

One hundred and one tea samples including green tea, dark tea, scented tea, black tea, and oolong tea were screened and confirmed for the contamination of 31 organochlorine pesticides (OCPs) and 19 pyrethroids (PYs) by gas chromatography-negative chemical ionization-mass spectrometry (GC-NCI-MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS). 50 pesticides, 3 deuterium-labeled PYs, and 24 (13)C-labeled OCPs were separated well with the limits of detection (LODs) ranging from 0.02 to 4.5 µg/kg for GC-NCI-MS, and the positive samples were verified by GC-MS/MS with LODs of 0.1-5.0 µg/kg. High detection rates for some PYs, such as 63.4% for bifenthrin (not detected (ND)-3.848 mg/kg), 55.4% for λ-cyhalothrin (ND-3.244 mg/kg), 46.5% for cypermethrin (ND-0.499 mg/kg), and 24.8% for fenvalerate (ND-0.217 mg/kg), were found in the 101 tea samples. Endosulfan, DDTs, HCHs, and heptachlor, the persistent OCPs, were frequently detected with rates of 63.4% (ND-1.802 mg/kg), 56.4% (ND-0.411 mg/kg), 24.8% (ND-0.377 mg/kg), and 15.8% (ND-0.100 mg/kg), respectively.

[Determination of 11 industrial antioxidants in the aqueous simulants by ultra-performance liquid chromatography-tandem mass spectrometry].

An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed to identify and determine 11 industrial antioxidants in the aqueous simulants. A ProElut PLS SPE column was used for the enrichment, and an ACQUITY UPLC BEH C18 UPLC column (100 mm x 2.1 mm, 1.7 µm) was used for separation by the gradient elution with pure water and acetonitrile as the mobile phases. The MS/MS detection was performed with an electrospray ionization (ESI) source in negative mode. The external standard method was used for quantitation in the present study. The linear ranges of the 11 analytes were from 5.0 to 100 µg/L. The coefficients of correlation were greater than 0.995. The recoveries of blank aqueous simulants fortified with the 11 analytes at the levels of 5.0, 10.0 and 20.0 µg/L were 61.4% to 109.4% with the relative standard deviations varied from 3.9% to 18.2% (n = 6). The LODs and LOQs of the 11 analytes in aqueous simulants were 0.2-1.0 µg/L and 0.5-3.0 µg/L, respectively. This method is highly sensitive and accurate, and can be applied to the determination of the 11 trace industrial antioxidants in the aqueous simulants.

[Determination of beta2-agonists and beta-blockers in urine using high performance liquid chromatography-ion trap mass spectrometry].

A method has been developed for the determination of 23 beta2-agonists and 5 beta-blockers in urine samples using high performance liquid chromatography-ion trap mass spectrometry (HPLC-IT-MS). Urine samples were first deproteinized by high-speed frozen centrifugation, and the supernatants were loaded on an Extrelut diatomite column for clean-up. The analytes were eluted by ethyl acetate and concentrated for further analysis. The analytical separation was performed on an AtlantisT3-150 mm chromatographic column with the gradient elution using methanol and water (containing 0.1% formic acid). The detection was carried on a linear ion trap mass spectrometer under multiple reaction monitoring (MRM) mode with the source operated in positive mode of electrospray ionization (ESI+). Nine deuterium labeled beta2-agonists were used as internal standards for quantitative analysis. The results showed that the linear ranges for 23 beta2-agonists and 5 beta-blockers were 0.005-0.16 mg/L, and the limits of detection were all around 0.2 microg/L. The mixed standard solution was added into the blank urine samples, and the recoveries of 23 beta2-agonists and 5 beta-blockers were ranged from 57.1% to 127.7% with the relative standard deviations of 1.1%-31.1%. The results demonstrate that the method is easy, fast, sensitive, and suitable for the confirmation and quantification of 23 beta2-agonists and 5 beta-blockers in urine samples.