Specific recognition of protein by deep eutectic solvent-based magnetic ß-cyclodextrin molecularly imprinted polymer.

A magnetic ß-cyclodextrin (MCD) surface molecularly imprinted polymer (MIP) based on deep eutectic solvents (DESs) as cross-linker and functional monomer (MCD@DES-MIP) was successfully synthesized for the specific recognition of bovine hemoglobin (BHb). The adsorption behavior of MCD@DES-MIP for BHb was investigated by adsorption thermodynamics, adsorption kinetics, and pH control experiments. The maximum adsorption capacity of MCD@DES-MIP for BHb under the optimized conditions was 195.94 mg g-1 and the imprinting factor was 4.68. In addition, the competitive adsorption experiments demonstrated that MCD@DES-MIP showed excellent selective extraction ability for BHb in the binary mixture of BHb and bovine serum albumin (BSA). The actual sample analysis manifested that MCD@DES-MIP effectively separated BHb from complex samples. The results of circular dichroism spectra proved that the secondary structure of BHb did not change during elution. The result indicated that MCD@DES-MIP can be used as a new imprinting material for the separation and purification of BHb.Graphical abstract Magnetic imprinted microspheres (MCD@DES-MIP) were prepared by free radical polymerization using magnetic ß-cyclodextrin (MCD) as carrier, deep eutectic solvents (DESs) as functional monomer and cross-linker. MCD@DES-MIP show high adsorption capacity and excellent selectivity for BHb.

Knockdown of Trop2 inhibits proliferation and migration and induces apoptosis of endometrial cancer cells via AKT/ß-catenin pathway.

Endometrial cancer (EC) is the most common gynaecologic malignancy in western countries and has been reported to account for about 7% of female malignant tumours and 20% to 30% of female genital system malignant tumours. Accumulating evidence showed the expression of human trophoblast cell surface antigen 2 (Trop2) was abnormal in many cancers; however, the expression and role of Trop2 in EC are not clear. The Trop-2 protein expression was detected by western blot in EC cells. Cell proliferation, apoptosis, and migration were measured by CCK-8, flow cytometry, and Transwell assay, respectively. The epithelial mesenchymal transition (EMT) and AKT/ß-catenin signalling pathway-related proteins in EC cell lines were detected by western blot assay following Trop2 gene silencing. The present study revealed that the Trop2 protein was highly expressed in EC cell lines compared with human endometrial epithelial cells. The Trop2 mRNA and protein were obviously decreased following transfection with Trop2-siRNA sequence in KLE and Ishikawa cells. Meanwhile, Trop2 gene silencing in KLE and Ishikawa cells strongly inhibited cell proliferation and migration and increased cell apoptosis. Investigation into the molecular mechanism indicated that the Trop2 gene silencing suppressed EMT and AKT/ß-catenin signalling pathway activation. SIGNIFICANCE OF THE STUDY: These findings suggested that Trop2 silencing inhibited EC cell proliferation and migration and promoted cell apoptosis. The mechanism might be related to the inhibition of the AKT/ß-catenin signalling pathway in EC cells. Therefore, Trop2 may be a potential therapeutic target for the treatment of EC.

Screening of differentially expressed proteins from syncytiotrophoblast for severe early-onset preeclampsia in women with gestational diabetes mellitus using tandem mass tag quantitative proteomics.

BACKGROUND: Previous studies have revealed that women with gestational diabetes mellitus (GDM) have an increased risk of developing preeclampsia (PE). The possible reason is the abnormal lipid metabolism caused by GDM that leads to dysfunction of vascular endothelial cells and atherosclerosis, resulting in the onset of PE. However, studies focusing on the pathogenesis of PE in syncytiotrophoblast of GDM patients are lacking. This study aimed to compare differentially expressed proteins from syncytiotrophoblast between women with GDM and women with GDM with subsequently developed PE. METHODS: Syncytiotrophoblast samples were obtained from pregnant women immediately after delivery. To explore the protein expression changes of syncytiotrophoblast that might explain the pathogenesis of PE in women with GDM, quantitative proteomics was performed using tandem mass tag (TMT) isobaric tags and liquid chromatography-tandem mass spectrometry. Bioinformatics analysis was performed to enrich the biological processes that these differentially expressed proteins were involved in. RESULTS: A total of 28,234 unique peptides and 4140 proteins were identified in all samples. Among them, 23 differentially expressed proteins were identified between patients with GDM and patients with GDM with subsequently developed PE. Therein, 11 proteins were upregulated and 12 proteins were downregulated. Two relative proteins (FLT1 and PABPC4) were independently verified using immunoblotting analysis. Bioinformatic results indicated that the onset of PE in patients with GDM is a multifactorial disorder, involving factors such as apoptosis, transcriptional misregulation, oxidative stress, lipid metabolism, cell infiltration and migration, and angiogenesis. CONCLUSION: These results indicated that the inadequacy of endometrium infiltration, angiogenic disorder, and oxidative stress in syncytiotrophoblast are more likely to occur in patients with GDM and may be the potential mechanisms leading to such patients secondarily developing severe early-onset PE.

A green deep eutectic solvent modified magnetic titanium dioxide nanoparticles for the solid-phase extraction of chymotrypsin.

Magnetic titanium dioxide nanoparticles modified with green deep eutectic solvent (DES) composed of choline chloride (ChCl) and xylitol (Xyl) (Fe3O4@TiO2@[ChCl][Xyl]) were synthesized and applied to the solid-phase extraction(MSPE) of chymotrypsin (Chy). The physicochemical properties and morphology of Fe3O4@TiO2@[ChCl][Xyl] was characterized by Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), Zeta potential, X-ray diffraction (XRD), vibrating sample magnetometer (VSM) and transmission electron microscope (TEM). The experiment parameters such as initial concentration of Chy, extraction time, pH value, ionic strength, extraction temperature and sample matrix were effectively optimized. Under the optimal experimental conditions, the extraction capacity of Fe3O4@TiO2@[ChCl][Xyl] obtained a significantly improvement after the modification of Fe3O4@TiO2 nanoparticles by [ChCl][Xyl], and reached up to 347.8 mg g-1. In the elution experiment, 10% sodium dodecyl sulfate-acetic acid (SDS-HAc) was used as eluent, achieving an elution rate of 85.9% for the Chy on Fe3O4@TiO2@[ChCl][Xyl]. And the Fe3O4@TiO2@[ChCl][Xyl] still maintained a good extraction capacity for Chy after six times of reuse. The application result in the extraction of Chy from porcine pancreas crude extract showed a good practical application ability for Chy extraction. All the results indicated that the synthesized Fe3O4@TiO2@[ChCl][Xyl] has good application potential in the extraction of biomolecular molecules such as protein.

A novel"turn-off"fluorescence assay based on acid-copper nanoclusters in deep eutectic solvent micelles for co-aggregation inducing fluorescence enhancement and its application.

As mixtures, deep eutectic solvent (DES) is designability. By adjusting the long alkyl chain hydrogen bond acceptors (HBAs) or hydrogen bond donors (HBDs), the DES displays surfactant characteristics and can form micelles. Hence, a novel, simple, facile and green natural organic acids capped copper nanoclusters (Aci-CuNCs) was synthesized and the spectrum behavior of Aci-CuNCs in DES micelles was researched. It was found that the surfactant-like DES can form micellar co-aggregation with Aci-CuNCs, resulting in the fluorescence (FL) intensive of Aci-CuNCs increase. Corresponding performance of spectral properties of Aci-CuNCs in DES medium were systematically studied by fourier transform infrared spectrometer, 3D FL spectroscopy, FL emission/excitation spectra, ultraviolet absorption spectroscopy. In the mechanism exploration part, on the one hand, the existence of micellar co-aggregation was confirmed by the conductivity, the mass effect of DES, dynamic light scattering and transmission electron microscopy. On the other hand, the influence of different kinds of DESs (types of HBAs/HBDs, molar ratio) and some possible factors (ionic strength and temperature) were discussed in detail to investigate the main driving forces for the formation of micellar co-aggregates. The results of mechanism exploration prove that the long alkyl chain of DES is amphiphilic which can form micellar co-aggregation with Aci-CuNCs through hydrogen bonding. The DES micelle provides Aci-CuNCs with a relatively stable and closed micro-environment which can effectively prevent collisions with water molecules and weakening of fluorescence intensity. On the basic of the above research, a "turn-off" fluorimetric method based on Aci-CuNCs in DES medium was applied for the determination of Fe3+. Under the optimum conditions, the assay worked in the Fe3+ concentration ranges from 1 to -20 µM and had a detection limit of 0.0374 µM. Method validation study illustrates the proposed system can provide a good accuracy, repeatability and stability conditions. Furthermore, the real sample analysis result demonstrates that no obvious matrix effect is found. As a consequence, the FL assays (Aci-CuNCs-based DES) composed of natural organic acid capped CuNCs and green solvent DES provides a simple, gentle and environmentally friendly method for the detection of iron ions.

A deep eutectic solvent modified magnetic ß-cyclodextrin particle for solid-phase extraction of trypsin.

An environmentally friendly deep eutectic solvent (DES) composed of trifluoroacetamide and benzyltrimethylammonium chloride modified ß-cyclodextrin (CD) grafted magnetic beads (MB-NH2@CD@DES) were synthesized for the first time and applied to the solid-phase extraction of trypsin. Among the five trypsin extractants prepared in this work (MB, MB-NH2, MB-NH2@CD, MB-NH2@DES, MB-NH2@CD@DES), the extractant MB-NH2@CD@DES with higher extraction capacity for trypsin was selected as final extractant. The extraction capacity of MB-NH2@CD@DES for trypsin can reach up to 549.87 mg⋅g-1 under the optimized conditions. The Langmuir adsorption equilibrium was found fitted better with equilibrium relation between MB-NH2@CD@DES and trypsin than Freundlich adsorption equilibrium. And a superior extraction for trypsin was verified by comparing the extraction capacity of MB-NH2@CD@DES for trypsin and four other common proteins. Compared with some reported trypsin extractants, the MB-NH2@CD@DES had a shorter extraction process, higher extraction capacity, more convenient operation of separation, a safer and more environmentally friendly synthesis process. With the optimized eluent, a great elution rate (74.32%) of trypsin was achieved. The absolute recovery of trypsin in trypsin standard solution was calculated to be 16.8%. And the extraction capacity of MB-NH2@CD@DES toward trypsin still maintained well after ten times recycling and reuse. The detection limit (LOD) and quantitative limit (LOQ) were 0.072 mg⋅mL-1 and 0.240 mg⋅mL-1 respectively. By sodium dodecyl sulfate polyacrylamide gel electrophoresis experiment, the extraction ability of MB-NH2@CD@DES to trypsin from real sample was fully demonstrated. All above results showed the potential of fabricated MB-NH2@CD@DES as a superior extractant for trypsin from real complex samples.

A new magnetic molecularly imprinted polymer based on deep eutectic solvents as functional monomer and cross-linker for specific recognition of bovine hemoglobin.

A novel magnetic molecularly imprinted polymer only based on deep eutectic solvents (DESs-MMIP) was successfully synthesized. The DESs-MMIP was constructed by using 2-hydroxyethyl methacrylate/tetrabutylammonium chloride deep eutectic solvent (DES1) as functional monomer, arylamide/(3-acrylamidopropyl) trimethylammonium chloride deep eutectic solvent (DES2) as cross-linker and bovine hemoglobin (BHb) as template through surface imprinting technology. The obtained DESs-MMIP was characterized by transmission electron microscope, X-ray diffraction, fourier transform infrared spectrometry, thermal gravimetric analysis and vibrating sample magnetometer. Under the optimized conditions, the maximum adsorption capacity of DESs-MMIP on BHb was 229.54 mg g-1 and the imprinting factor reached up to 21.89. The selective adsorption experiments indicated that compared with seven references, DESs-MMIP showed significant selectivity for BHb. The new-type DESs-MMIP exhibited higher adsorption capacity and imprinting factor on BHb than molecularly imprinted polymers constructed with traditional functional monomer and cross-linker in reported methods. The recognition of BHb by DESs-MMIP in calf blood samples demonstrated the practicality of the particles. The DESs-MMIP only based on deep eutectic solvents with excellent selectivity is expected to become an ideal candidate for selective recognition of BHb in complicated samples.

The association between calcium supplement and preeclampsia and gestational hypertension: a systematic review and meta-analysis of randomized trials.

OBJECTIVES: The purpose of this study is to investigate whether calcium supplement with or without other drugs could reduce the risk of preeclampsia and gestational hypertension based on existed evidence, and to clarify whether there is discrepant effect among different population and using different dose. METHODS: PubMed, Cochrane library, and EMBASE database were searched. Two authors independently screened all records and extracted data. The meta-analysis was performed to calculate risk ratios and 95% CIs using random-effects models. RESULTS: 27 studies, with 28 492 pregnant women were included. The results showed calcium supplement was associated with lower incidence of preeclampsia (RR 0.51, 95% CI: 0.40 to 0.64) and gestational hypertension (RR 0.70, 95% CI: 0.60 to 0.82). Sub-analyses revealed high-dose (1.2-2 g/day), moderate-dose (0.6-1.2 g/day), and low-dose (<0.6 g/day) of calcium supplement could reduce the risk of preeclampsia. For gestational hypertension, only high dose and moderate dose groups were associated with reducing the risk of gestational hypertension. However, we could draw a conclusion which does group was the most protective, as we were unable to directly compare the effects of different doses. CONCLUSIONS: This study indicated calcium supplementation might decrease the risk of preeclampsia and gestational hypertension. And results of subgroups analyses enhanced our confidence to the protective effect of calcium supplementation. However, further studies with direct comparison of different dose of calcium supplementation are needed to explore the ideal dose of calcium supplementation to prevent preeclampsia and gestational hypertension.

A New Pansharpening Method Based on Spatial and Spectral Sparsity Priors.

The development of multisensor systems in recent years has led to great increase in the amount of available remote sensing data. Image fusion techniques aim at inferring high quality images of a given area from degraded versions of the same area obtained by multiple sensors. This paper focuses on pansharpening, which is the inference of a high spatial resolution multispectral image from two degraded versions with complementary spectral and spatial resolution characteristics: a) a low spatial resolution multispectral image; and b) a high spatial resolution panchromatic image. We introduce a new variational model based on spatial and spectral sparsity priors for the fusion. In the spectral domain we encourage low-rank structure, whereas in the spatial domain we promote sparsity on the local differences. Given the fact that both panchromatic and multispectral images are integrations of the underlying continuous spectra using different channel responses, we propose to exploit appropriate regularizations based on both spatial and spectral links between panchromatic and the fused multispectral images. A weighted version of the vector Total Variation (TV) norm of the data matrix is employed to align the spatial information of the fused image with that of the panchromatic image. With regard to spectral information, two different types of regularization are proposed to promote a soft constraint on the linear dependence between the panchromatic and the fused multispectral images. The first one estimates directly the linear coefficients from the observed panchromatic and low resolution multispectral images by Linear Regression (LR) while the second one employs the Principal Component Pursuit (PCP) to obtain a robust recovery of the underlying low-rank structure. We also show that the two regularizers are strongly related. The basic idea of both regularizers is that the fused image should have low-rank and preserve edge locations. We use a variation of the recently proposed Split Augmented Lagrangian Shrinkage (SALSA) algorithm to effectively solve the proposed variational formulations. Experimental results on simulated and real remote sensing images show the effectiveness of the proposed pansharpening method compared to the state-of-the-art.