Nucleic acid aptasensor with magnetically induced self-assembly for the detection of EpCAM glycoprotein.

A "turn-on" aptasensor for label-free and cell-free EpCAM detection was constructed by employing magnetic α-Fe2O3/Fe3O4@Au nanocomposites as a matrix for signal amplification and double-stranded complex (SH-DNA/Apt probes) immobilization through Au-S binding. α-Fe2O3/Fe3O4@Au could be efficiently assembled into uniform and stable self-assembly films via magnetic-induced self-assembly technique on a magnetic glassy carbon electrode (MGCE). The effectiveness of the platform for EpCAM detection was confirmed through differential pulse voltammetry (DPV). Under optimized conditions, the platform exhibited excellent specificity for EpCAM, and a strong linear correlation was observed between the current and the logarithm of EpCAM protein concentration in the range 1 pg/mL-1000 pg/mL (R2 = 0.9964), with a limit of detection (LOD) of 0.27 pg/mL. Furthermore, the developed platform demonstrated good stability during a 14-day storage test, with fluctuations remaining below 93.33% of the initial current value. Promising results were obtained when detecting EpCAM in spiked serum samples, suggesting its potential as a point-of-care (POC) testing.

Electrochemical peptide nucleic acid functionalized α-Fe2O3/Fe3O4 nanosheets for detection of CYP2C19*2 gene.

The CYP2C19*2 gene carriers and non-carriers are closely related to the dosage of clopidogrel. To correctly guide the use of clopidogrel and promote individualized therapy, an ultra-sensitive electrochemical biosensor was developed for the detection of CYP2C19*2 gene. The heterogeneous α-Fe2O3/Fe3O4 nanosheets were prepared via the hydrothermal-calcination process, and the preparation parameters were optimized. The average diameter and thickness of the nanosheets were approximately 150 nm and 53 nm, respectively; and the saturation magnetization was 80.2 emu/g. The α-Fe2O3/Fe3O4@Au nanosheets were prepared by sodium borohydride reduction method, and self-assembled to the electrode surface with magnetic field. Ultra-sensitive detection of CYP2C19*2 gene was realized through the recognition ability of strong single base mismatching of peptide nucleic acid and signal amplification effect of magnetic α-Fe2O3/Fe3O4@Au nanosheets. Under optimal detection conditions, the current had a good linear correlation with the negative logarithm of CYP2C19*2 gene concentration in the range 1 pM-1 nM, and the detection limit was 0.64 pM (S/N = 3). Meanwhile, the electrochemical signals of target DNA and incomplete complementary DNA were detected. The constructed biosensor exhibited good selectivity, reproducibility, and stability, providing a promising strategy for the detection of other gene mutations by electrochemical biosensors.

Symmetry dual functional pyrimidine-BODIPY probes for imaging targeting and activity study.

Nondestructive diagnosis of tumor has always been the goal of scientists. Fluorescent dyes have become the rising star in the field of cancer diagnosis because of their excellent characteristics. Therefore, in this work, fluorescence probes d-Y-B and dO-Y-B with anti-tumor activity were constructed by introducing pyrimidine groups with high anti-tumor activity using fluorescence dye BODIPY as parent nucleus. The modified BODIPY group in the structure had the advantage of fluorescent dye, ensuring the strong fluorescence and photosensitivity of the target compound. That ethylenediamine acts as a bridge with two -NH- groups to increase molecular hydrogen bonding, and can bind firmly to multiple proteins. Co-localization of the target compounds d-Y-B and dO-Y-B with the hoechst dye for labeling living cells showed that these compounds had high biocompatibility and photostability for localization to HeLa cells. In vivo imaging in mice can realize specific localization and real-time visualization of tumor cells. The results of cytotoxicity experiments in vitro and computer software simulating molecular docking confirmed the potential of the target compounds as an anticancer agents. The bifunctional probe realized visualization of cancer cells in mice, and can kill cancer cells by anti-proliferation, which may provide a direction for future anticancer drug development.

Label-free electrochemical aptasensor based on magnetic α-Fe2O3/Fe3O4 heterogeneous hollow nanorods for the detection of cancer antigen 125.

A label-free electrochemical aptasensor based on magnetic α-Fe2O3/Fe3O4 heterogeneous hollow nanorods was developed for the detection of cancer antigen 125 (CA125). Magnetic α-Fe2O3/Fe3O4 heterogeneous hollow nanorods were successfully prepared by the hydrolysis-calcination method, functionalized with polyethyleneimine (PEI), and modified with HAuCl4 to form magnetic α-Fe2O3/Fe3O4-Au nanocomposites with a layer of 5 nm gold nanoparticles (Au NPs) on the surface. The magnetic α-Fe2O3/Fe3O4-Au nanocomposites were used to fix the DNA-aptamer probe to amplify the current signal. The intensity of the current signal was proportional to the concentration of CA125, indicating that the sensor was a turn-on sensor. The linear range of the electrochemical aptasensor was 5-125 U/mL (R2 = 0.9975), and the limit of detection (LOD) was 2.99 U/mL. The electrochemical aptasensor exhibited favorable specificity, reproducibility, and stability. The analytical performance of the aptasensor in real serum samples was also investigated.

Meta-analysis of the association between sclerostin level and adverse clinical outcomes in patients undergoing maintenance haemodialysis.

BACKGROUND: Studies regarding the relationship of sclerostin (Scl) with clinical outcomes in patients undergoing maintenance haemodialysis have yielded controversial findings. This meta-analysis was performed to investigate the predictive role of Scl in this patient population. METHODS: Several electronic medical databases (e.g. PubMed, Embase, Web of Science and Cochrane Library) were searched for eligible studies through December 20, 2019. Summary hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated based on Scr level (high or low) using a random or fixed effects model. RESULTS: From among 641 initially screened publications, 16 eligible studies were included in this meta-analysis. A high Scl level was not associated with cardiovascular events [HR = 0.8 (95% CI, 0.42-1.35)] or all-cause mortality [HR = 0.93 (95% CI, 0.56-1.54)]. There was high heterogeneity, but no evidence of publication bias. Interestingly, a high Scl level was associated with reduced cardiovascular events [HR = 0.44 (95% CI, 0.29-0.69)] in the subgroup by shorter follow-up period or all-cause mortality [pooled HR = 0.58 (95% CI, 0.36-0.91)] by shorter dialysis vintage. CONCLUSION: This meta-analysis indicated that a high Scl level did not predict total clinical outcomes in patients undergoing maintenance haemodialysis despite survival benefits in the subgroups. The predictive role of Scl in these patients should be further evaluated in large prospective studies.

Nano-Graphene Enclosed Multi Nitrogen: Dynamic Hierarchical Self-Assemble Property for Lithium Ion Storage.

Led to significant capacity improvement to 1800 mA/h after 100 cycles for nano-graphene-N4, which is the first report for a carbonaceous materials anode. In addition, the doping level, id est, number of nitrogen atoms, had a significant influence on the molecular self-assembled structures through hierarchical self-assembly. As the nitrogen concentration increased, the d-space between the nanosheets increased from 3.4 to 4.3. The capacity of the nano-graphene increased greatly from 500 mAh/g for nano-graphene without N-doping to 1800 mAh/g for nano-graphene with nanographene-N4, indicating that the capacity is related to the structures, and was defined and the relationship between performance and structure was determined.

A Novel Process for the Preparation of ZnO Nanoparticles and ZnO/Ni0.5Zn0.5Fe2O4 Nanocomposites.

A novel solution combustion and gel calcination process for the preparation of ZnO nanoparticles and ZnO/Ni0.5Zn0.5Fe2O4 nanocomposites was introduced. The phase identification, morphologies and magnetic properties of ZnO nanoparticles and ZnO/Ni0.5Zn0.5Fe2O4 nanocomposites were confirmed by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscope (TEM), vibrating sample magnetometer (VSM). The results showed that the concentration of zinc nitrate and the calcination temperature were two key factors for the grain size, crystallinity and properties of ZnO nanoparticles, while, the mass ratio of ZnO and Ni0.5Zn0.5Fe2O4 in ZnO/Ni0.5Zn0.5Fe2O4 nanocomposites was the important factor for the specific magnetizations (Ms) of ZnO/Ni0.5Zn0.5Fe2O4 nanocomposites.

1,25(OH)2D3 Activates Autophagy to Protect against Oxidative Damage of INS-1 Pancreatic Beta Cells.

Diabetes mellitus is a serious disease endangering human health worldwide. Vitamin D (Vit D) is a well-characterized regulator of calcium-phosphorus metabolism that also exerts other biological effects extending far beyond mineral homeostasis. Some epidemiological studies have suggested that Vit D has a role in defense against diabetes, although the mechanism remains unclear. Autophagy, an intracellular catabolic process, is necessary to maintain the normal structure and function of host cells. In our previous study, we found that Vit D could induce autophagy of pancreatic beta cells and prevent insulitis, although the underlying mechanisms remain to be fully elucidated. In this study, the protective effect of 1,25(OH)2D3, the physiologically active metabolite of Vit D, against streptozotocin-induced cytotoxicity in rat insulinoma cell line (INS-1) cells was explored. Cell viability and insulin secretion of INS-1 cells in response to different treatments were measured with a cell counting kit and enzyme-linked immune absorbent assay (ELISA), respectively. In addition, malondialdehyde (MDA) content and total antioxidant capacity (T-AOC) were measured by ELISA. RT-PCR and Western blot analyses were used to detect autophagy levels, reactive oxygen species (ROS) was assessed by fluorescence microscope, ultrastructure analysis was performed using transmission electron microscopy. The results demonstrated that 1,25(OH)2D3 could increase cell viability and insulin secretion of INS-1 cells, and protected cells from oxidative damage induced by streptozotocin (STZ) through autophagy activation. These findings shed light on mechanisms underlying the ameliorative effects of Vit D on diabetes mellitus.

Klotho Inhibits Unilateral Ureteral Obstruction-Induced Endothelial-to-Mesenchymal Transition via TGF-ß1/Smad2/Snail1 Signaling in Mice.

This study aimed to evaluate repression of Klotho on unilateral ureteral obstruction (UUO)-induced renal fibrosis and endothelial-to-mesenchymal transition (EndoMT) in mice. Renal fibrosis model was established by UUO in C57BL/6 male mice. Recombinant Klotho protein was administered to UUO mice as treatment group, and the mice in sham and UUO group were administered with an equal volume of vehicle. EndoMT biomarkers and TGF-ß1/Smad2/Snail1 signaling were examined by immunofluorescence, immunohistochemistry, and western blotting assays. UUO deteriorated kidney function and resulted in increased expression of the mesenchymal marker α-smooth muscle actin and decreased expression of vascular endothelial cadherin, an endothelial marker. Moreover, UUO enhanced TGF-ß1, phosphorylated Smad2 (p-Smad2), and Snail1 expression. Interestingly, Klotho treatment suppressed UUO-induced TGF-ß1, p-Smad3, and Snail1 expression, which was accompanied by alleviation of the EndoMT process. Our findings demonstrated that Klotho significantly ameliorated EndoMT progression by targeting TGF-ß1/Smad/Snail1 signaling in UUO mice, which provides the possibility for Klotho-based therapeutic protection against renal fibrosis.

Adsorption Performance of Congo Red onto Magnetic Ni0.4Co0.2Zn0.4Fe2O4 Nanoparticles Prepared via the Rapid Combustion Process.

Magnetic Ni0.4Co0.2Zn0.4Fe2O4 nanoparticles were prepared via the rapid combustion process, the morphology and magnetic property of them were investigated by SEM, TEM, XRD, VSM, and FTIR. The as-prepared magnetic Ni0.4Co0.2Zn0.4Fe2O4 nanoparticles calcined at 400 °C for 2 h with absolute alcohol of 20 mL were characterized with the average nanoparticle size of around 25 nm and the specific magnetization of 35.2 emu/g. The adsorption kinetics and adsorption isotherms of Congo red (CR) from aqueous solutions onto Ni0.4Co0.2Zn0.4Fe2O4 nanoparticles were investigated by UV spectroscopy at room temperature; the adsorption kinetics data were in good agreement with the pseudo-second-order kinetic model in the initial CR concentrations of 200-500 mg/L. By comparison of Langmuir, Freundlich and Temkin models for adsorption isotherms of CR, the Temkin model (correlation coefficient R2 = 0.9969) could be used to evaluate the adsorption isotherm of CR onto the magnetic Ni0.4Co0.2Zn0.4Fe2O4 nanoparticles at room temperature, which suggested that the adsorption of CR onto the magnetic Ni0.4Co0.2Zn0.4Fe2O4 nanoparticles was a hybrid of monolayer and multilayer absorbing mechanism.

Dynamic Hierarchical Self-Assemble Small Molecule Structure Hexabenzocoronene for the High-Performance Anodes Lithium Ion Storage.

This study examined the characteristics of small molecular structure nano-graphene in a dynamic hierarchical self-assembly and found that graphene is rearranged under its own pressure during dynamic aggregation and water ripples are formed by the d-spacing. The composition and structure were studied using a range of material characterization techniques. No covalent bonds were observed between molecules, and the self-assembled driving force was the only intermolecular interaction: Van der Waals' force in the intra-layer and π-π interactions between layers. The arranged-rearranged structures provided a range of lithium ion shuttle channels, including the space between layers and diffusing through the nanosheets, which decrease the diffusion distance of lithium ions remarkably and reduce the irreversible capacity of the battery.

Ginsenoside-Rg1 Protects against Renal Fibrosis by Regulating the Klotho/TGF-ß1/Smad Signaling Pathway in Rats with Obstructive Nephropathy.

Ginsenoside-Rg1 (G-Rg1) is an agent isolated from Panax ginseng that exerts anti-fibrotic effects; however, the mechanism is still unclear. Herein, we investigated whether G-Rg1 administration can mitigate or reverse unilateral ureteral obstruction (UUO)-induced renal fibrosis by regulating the Klotho/transforming growth factor (TGF)-ß1/Smad signaling pathway in rats. Sprague-Dawley male rats were subjected to UUO, and rats in the treatment group were administered G-Rg1 or G-Rg1 plus Klotho short hairpin RNA interference (shRNA), while rats in the control and model groups were administered vehicle for 14 d. Epithelial-mesenchymal transition (EMT) biomarkers and Klotho/TGF-ß1 signaling molecules were examined by immunohistochemistry, quantitative real-time PCR and Western blotting. Immunohistochemistry showed that UUO induced increased pro-fibrotic TGF-ß1 expression, overexpression of the mesenchymal marker, α-smooth muscle actin (α-SMA), and suppression of the epithelial marker, E-cadherin. Moreover, Western blotting analysis indicated that UUO promoted TGF-ß1 and phosphorylated Smad3 (p-Smad3) expression (p<0.01), but blocked Klotho and Smad7 expression (p<0.01). After G-Rg1 administration, the UUO-induced TGF-ß1 and p-Smad3 expression was suppressed (p<0.01), whereas the reduced Klotho and Smad7 expression was reversed (p<0.05), followed by amelioration of the EMT process. Intriguingly, the G-Rg1 effects were largely abrogated by Klotho knockdown. Furthermore, Klotho expression was upregulated by G-Rg1 treatment at the mRNA and protein levels. Our results suggest that G-Rg1 may be beneficial for ameliorating renal fibrosis by targeting Klotho/TGF-ß1/Smad signaling in UUO rats.

Adsorption Performance of DNA onto Magnetic Core-Shell Structure (1-x)Ni0.5Zn0.5Fe2O4/xSiO2 Nanopowders Prepared via Methanol Combustion Process.

Magnetic core-shell structure (1-x)Ni0.5Zn0.5Fe2O4/xSiO2 (x = 0-0.2) nanopowders were successfully prepared via the methanol combustion process, the as-prepared magnetic core-shell structure nanopowders were characterized by XRD, SEM, TEM, HRTEM, BET, and VSM techniques, and the adsorption performance of DNA onto them was investigated. The results showed that the magnetic core-shell structure 0.9Ni0.5Zn0.5Fe2O4/0.1SiO2 nanopowders exhibited a good adsorption capacity of DNA, and the adsorbance of DNA for magnetic core-shell structure 0.9Ni0.5Zn0.5Fe2O4/0.1SiO2 nanopowders was enhanced to 2.8 times for Ni0.5Zn0.5Fe2O4 nanoparticles. The adsorption kinetics of DNA in aqueous solution onto the magnetic core-shell structure 0.9Ni0.5Zn0.5Fe2O4/0.1SiO2 nanopowders at room temperature were investigated, and the kinetics data were in good agreement with the pseudo-first-order kinetic model in the initial DNA concentration of 50 µg/mL.

Adsorption Kinetics of Bovine Serum Albumin onto Ni0.5Zn0.5Fe2O4/SiO2 Nanocomposites Prepared via the Solution Combustion Process.

Magnetic Ni0.5Zn0.5Fe2O4/SiO2 nanocomposites were prepared via the solution combustion process, their structure and magnetic properties were investigated by XRD, SEM, TEM and VSM. The asprepared magnetic Ni0.5Zn0.5Fe2O4/SiO2 nanocomposites were characterized with the average grain size of about 20 nm, the specific magnetization of 31.6 Am2/kg, and the specific surface area of 117.0 m2/g. The adsorption kinetics of bovine serum albumin (BSA) from aqueous solution onto the magnetic Ni0.5Zn0.5Fe2O4/SiO2 nanocomposites at room temperature was investigated. The kinetics data related to the adsorption of BSA from aqueous solutions were in good agreement with the pseudo-second-order kinetics model in the initial concentrations of 0.2-0.8 mg/mL.

Plasma s-Klotho is related to kidney function and predicts adverse renal outcomes in patients with advanced chronic kidney disease.

To investigate whether the soluble Klotho (s-Klotho) level in patients with chronic kidney disease (CKD) is related to kidney function and whether a low s-Klotho level can predict adverse renal outcomes or CKD progression in patients with advanced CKD. 112 patients with CKD stages 3-5 and 30 healthy volunteers were enrolled. Blood samples were collected to measure serum creatinine, calcium, phosphorus, intact parathyroid hormone, and hemoglobin. s-Klotho and fibroblast growth factor 23 (FGF23) were determined by ELISA. We first conducted a cross-sectional study to investigate correlations between s-Klotho and estimated glomerular filtration rate (eGFR) and other parameters. Patients were then followed prospectively for 20.1±10.1 months according to s-Klotho median level until serum creatinine doubled, or initiation of renal replacement therapy, or death. s-Klotho levels inpatients with CKD were significantly lower than that in the control group. For patients with CKD, there were no differences in age distribution among subgroups. However, s-Klotho level differed significantly across CKD stages, and it was lower in the advanced CKD group compared with the moderate CKD group. Correlation analysis revealed that s-Klotho was positively associated with eGFR, but inversely associated with FGF23. During the follow-up of 20.1±10.1 months, patients with higher s-Klotho levels showed a reduced risk of kidney adverse outcomes, including serum creatinine doubling and initiation of renal replacement therapy. Cox regression analysis revealed that low s-Klotho was an independent risk factor for CKD progression. s-Klotho level was closely correlated with kidney function, further, low s-Klotho level could predict adverse kidney disease outcomes in patients with progressive CKD.

Cyclosporine A Suppresses the Activation of the Th17 Cells in Patients with Primary Sjögren's Syndrome.

Primary Sjögren's syndrome (pSS) is a common autoimmune disease involving abnormal Th17 activation. The aim of the current study was to investigate the immunosuppression effect of Cyclosporine A (Cys A), a potent immunosuppressor on the proliferation and activation of T cells, on the activation of Th17 cells. Blood samples from both inactive and active pSS patients as well as healthy controls were collected and serum and peripheral blood mononuclear cells (PBMCs) were collected and tested for IL-17 and RORγt expression. Subsequently, PBMCs were treated in vitro with Cys A in a series of doses and incubation time and the effect of Cys A on inhibiting Th17 activation was tested by measuring IL-17 and RORγt expression. IL-17 in both serum and PBMCs as well as RORγt in PBMCs from active pSS patients were significantly elevated on both the mRNA and protein levels comparing to those from both inactive pSS patients and healthy controlCys A in the final concentration of 80ng/ml and the treatment time of 24h showed strong inhibition effect on the expression of IL-17 and RORγt in PBMCs from active pSS patients. However, Cys A in various doses and incubation times did not show much impact on inhibiting IL-17 as well as RORγt expression in PBMCs from healthy donors and inactive pSS patients. Cys A possesses the capability in immunosuppressing the activation of Th17 cells, suggesting that Cys A may be a potential treatment for pSS and maybe other autoimmune diseases.

Tranilast attenuates TGF-ß1-induced epithelial-mesenchymal transition in the NRK-52E cells.

We previously reported that tranilast can halt the pathogenesis of chronic cyclosporine nephrotoxicity in rats via the transforming growth factor-ß (TGF-ß) /Smad pathway, an important signaling system involved in epithelial-mesenchymal transition (EMT), but the exact underlying cellular mechanisms are not yet clear. Thus, by selecting TGF-ß1-induced normal rat kidney proximal tubular epithelial cells (NRK-52E) as a model, we demonstrated potential modifying effect of tranilast on EMT-induced by TGF-ß1 in vitro. NRK-52E cells were incubated with the blank vehicle (Dulbecco's modified Eagle's medium and F-12 (DMEM/F12) added with 10% fetal bovine serum (FBS)), 10 ng/ml TGF-ß1 alone or together with 100, 200 or 400µM tranilast for 48 h after incubation in medium containing 1% FBS for 24 h. Cell morphological changes were observed to confirm occurrence of EMT. Protein expressions of two typical markers of EMT, E-cadherin and α-smooth muscle actin (α-SMA), were assessed by western blotting and flow cytometry, respectively. Our results showed that TGF-ß1 induced spindle-like morphological transition, the loss of E-cadherin protein and upregulation of expression of α-SMA. However, the TGF-ß1-produced changes in cellular morphology, E-cadherin and α-SMA were inversed by tranlilast in concentration-dependent manner. Our findings indicate that tranilast can directly inhibit EMT. Thus, it may be implied that regulation of EMT be the target to prevent renal tubulointerstitial fibrosis.

[Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells].

OBJECTIVES: To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells. METHODS: PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or downregulation of PIAS3 expression levels in TJ905 cells. After that, the invasive effects of TJ905 cells were measured by Transwell assay, and the expression of PIAS3, tissue inhibitor of metalloproteinases (TIMP)3, matrix metalloprotease (MMP)-2, and MMP-9 were identified by Western blot. RESULTS: In vitro transfection efficiency of plasmids and oligonucleotides were separately 85.3% ± 3.1% and 95.1% ± 2.9%. PIAS3 overexpression plasmid transfection in vitro could effectively improve the expression of PIAS3 protein in TJ905 cells and inhibit the invasion of TJ905 cells (P < 0.05), and cell penetration ratio reduced from 87.9% ± 9.3% to 37.3% ± 7.9% compared with control group, while it upregulated TIMP3 and downregulated MMP-2, MMP-9 protein expression (P < 0.05); PIAS3 siRNA transfection could inhibit the PIAS3 protein expression of TJ905 cells and promote the invasion of TJ905 cells (P < 0.05), and cell penetration ratio increased from 83.9% ± 7.1% to 93.2% ± 3.1% compared with control group, while it downregulated TIMP3 and upregulated MMP-2, MMP-9 protein expression (P < 0.05). CONCLUSION: PIAS3 expression is closely related to the invasion properties of glioma TJ905 cells.