A novel ß-cyclodextrin-assisted enhancement strategy for portable and sensitive detection of miR-21 in human serum.

Benefiting from our discovery that ß-cyclodextrin (ß-CD) could enhance the catalytic activity of invertase through hydrogen bonding to improve detection sensitivity, a highly sensitive and convenient biosensor for the detection of miR-21 was proposed, which is based on the simplicity of reading signals from a personal glucose meter (PGM), combined with self-assembled signal amplification probes and the performance of ß-CD as an enhancer. In the presence of miR-21, magnetic nanoparticle coupled capture DNA (MNPs-cDNA) could capture it and then connect assist DNA/H1-invertase (aDNA/H1) and self-assembled signal amplification probes (H1/H2) in turn. As a result, a "super sandwich" structure was formed. The invertase on MNPs-cDNA could catalyze the hydrolysis of sucrose to glucose and this catalytic process could be enhanced by ß-CD. The PGM signal exhibited a linear correlation with miR-21 concentration within the range of 25 pmol L-1 to 3 nmol L-1, and the detection limit was as low as 5 pmol L-1 with high specificity. Moreover, the recoveries were 103.82-124.65% and RSD was 2.59-6.43%. Furthermore, the biosensor was validated for the detection of miR-21 in serum, and the results showed that miR-21 levels in serum samples from patients with Diffuse Large B-Cell Lymphoma (DLBCL) (n = 12) were significantly higher than those from healthy controls (n = 12) (P < 0.001). Therefore, the ingenious combination of PGM-based signal reading, self-assembled signal amplification probes and ß-CD as an enhancer successfully constructed a convenient, sensitive and specific biosensing method, which is expected to be applied to clinical diagnosis.

Fluorescence quenching determination of tetracyclines based on the synergistic oxidation effect between Fe3O4nanoparticles and tetracyclines.

In recent years, tetracyclines (TCs) is a hot research topic. Herein, we report an interesting discovery using the complexation of oxytetracycline and metal ions. In this study, according to the properties of Fe3O4nanoparticles (Fe3O4NPs) as a nanoenzyme, it can be used to catalyze the oxidation of KI by H2O2to produceI3-,while at the same timeI3-binds to rhodamine 6G (Rh6G) to form a conjoined particle (Rh6G ∼ I3)n, leading to a decrease in the fluorescence intensity of Rh6G. However, in the presence of TCs, Fe3O4NPs have a synergistic effect with TCs, leading to enhanced catalytic activity, as well as better selectivity compared to the activity of other reducing enzymes. Consequently,the fluorescent signal based on a resonance scattering effect between Rh6G andI3-is dependent on the concentration of TCs, thus achieving highly facile and robust detection of TCs. The limits of detection (LOD) of the method were 20 nM, 10 nM and 40 nM for oxytetracycline(OTC), tetracycline(TC) and chlortetracycline(CTC), respectively. Most importantly, the method can be successfully applied to the detection of TCs in milk, eggs, and honey. The recoveries of spiked samples ranged from 83.11 to 118.95%. Thus, a stable, hands-on strategy for the detection of TCs is proposed, which has potential applications in the field of food safety and environmental protection.

The latest application progress of radiomics in prediction and diagnosis of liver diseases.

INTRODUCTION: Early detection and individualized treatment of patients with liver disease is the key to survival. Radiomics can extract high-throughput quantitative features by multimode imaging, which has good application prospects for the diagnosis, staging and prognosis of benign and malignant liver diseases. Therefore, this paper summarizes the current research status in the field of liver disease, in order to help these patients achieve personalized and precision medical care. AREAS COVERED: This paper uses several keywords on the PubMed database to search the references, and reviews the workflow of traditional radiomics, as well as the characteristics and influencing factors of different imaging modes. At the same time, the references on the application of imaging in different benign and malignant liver diseases were also summarized. EXPERT OPINION: For patients with liver disease, the traditional imaging evaluation can only provide limited information. Radiomics exploits the characteristics of high-throughput and high-dimensional extraction, enabling liver imaging capabilities far beyond the scope of traditional visual image analysis. Recent studies have demonstrated the prospect of this technology in personalized diagnosis and treatment decision in various fields of the liver. However, further clinical validation is needed in its application and practice.

Revealing the metabolic mechanism of dandelion extract against A549 cells using UPLC-QTOF MS.

Dandelion extract exhibits potential anticancer activity and is expected to be a new type of natural anticancer drug. However, the effect mechanism of dandelion extract to lung cancer cells is still unclear. Here, untargeted metabolomics approach based on LC-MS was used to characterize the metabolic responses of A549 cells to dandelion extract exposure and to provide new clues for the antitumor mechanism of dandelion extract from the metabolomics perspective. A total of 16 differentially expressed and time-related metabolites were identified between dandelion extract exposure and control groups. The perturbed metabolic pathways of A549 cells after dandelion extract exposure mainly include the glycerophospholipid metabolism and purine metabolism. These results concluded that dandelion extract may exert anticancer activity by affecting malignant proliferation, disturbing the stability of cell membrane structure, reducing the adhesion of tumor cells to extracellular matrix and fibronectin, and finally inducing tumor cell death.

Discovery of lipid profiles of type 2 diabetes associated with hyperlipidemia using untargeted UPLC Q-TOF/MS-based lipidomics approach.

The incidence of type 2 diabetes (T2D) is rising rapidly and has become an important public health problem. According to reports, people with T2D often have hyperlipidemia. Hence, in the current study, a plasma non-targeted lipidomics method was used to study the differences in lipid profile between 36 T2D-associated hyperlipidemia patients and 43 healthy controls by ultra-performance liquid chromatography coupled with quadrupole time-of-flight high-definition mass spectrometry (UPLC Q-TOF/MS). Furthermore, we studied the differences in lipid profile between 36 T2D-associated hyperlipidemia patients and 41 T2D patients. Principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), S-plot and heatmap were used to analyze the lipid changes between the groups. Compared with the healthy control group, 37 lipids were significantly altered in the T2D-associated hyperlipidemia group, and when compared with the T2D group, 22 lipids were significantly altered in the T2D-associated hyperlipidemia group. Of all the detected lipids categories which included sphingolipids, glycerolipids, glycerophospholipids, prenol lipids and saccharolipids, glycerophospholipids accounted for the largest proportion in the two groups. Also, this study found that glycerophospholipid metabolism pathway was the most relevant pathway for these lipid metabolisms. The identified lipids may enhance the disease prediction and provide a new tool to monitor the progression of T2D-associated hyperlipidemia.

The association between soy-based food and soy isoflavone intake and the risk of gastric cancer: a systematic review and meta-analysis.

Soy contains many bioactive phytochemicals, such as isoflavones, which have the effect of preventing many cancers. Some studies have shown the beneficial effect of soy-based food and isoflavone intake on gastric cancer (GC), while others claimed no effect. Therefore, whether the beneficial effect of soy-based food is related to its fermentation or whether its protective effect comes from isoflavones still remains inconclusive. Our aim was to investigate the relationship between total soybean, fermented soybean, non-fermented soybean and isoflavone intake, and the risk of GC. Ten cohort studies and 21 case-control studies involving 916 354 participants were included. The association between soy-based food and isoflavone intake and the risk of GC was calculated with the pooled relative risks (RRs) for the highest versus lowest intake categories. The results showed that isoflavone intake might be a protective factor to GC, but the result was not statistically significant (RR = 0.92; 95% CI: 0.79-1.07). However, total soybean intake could significantly decrease the risk of GC by 36% (RR = 0.64; 95% CI: 0.51-0.80), which might be credited to non-fermented soybean products (RR = 0.79; 95% CI: 0.71-0.87). In contrast, high intake of fermented soybean products could increase the risk of GC (RR = 1.19; 95% CI: 1.02-1.38). High intake of total soybean and non-fermented soybean products could reduce the risk of GC, and high intake of fermented soybean products could increase the risk, which indicated that the beneficial effect of soy-based food might be related to its non-fermentation. However, high intake of isoflavones may not be associated with the incidence of GC. © 2021 Society of Chemical Industry.

In situ detection of plasma exosomal microRNA for lung cancer diagnosis using duplex-specific nuclease and MoS2 nanosheets.

MicroRNAs (miRNAs) encapsulated in tumor-derived exosomes are becoming ideal biomarkers for the early diagnosis and prognosis of lung cancer. However, the accuracy and sensitivity are often hampered by the extraction process of exosomal miRNA using traditional methods. Herein, this study developed a fluorogenic quantitative detection method for exosomal miRNA using the fluorescence quenching properties of molybdenum disulfide (MoS2) nanosheets and the enzyme-assisted signal amplification properties of duplex-specific nuclease (DSN). First, a fluorescently-labeled nucleic acid probe was used to hybridize the target miRNA to form a DNA/RNA hybrid structure. Under the action of the DSN, the DNA single strand in the DNA/RNA hybrid strand was selectively digested into smaller oligonucleotide fragments. At the same time, the released miRNA target triggers the next reaction cycle, so as to achieve signal amplification. Then, MoS2 was used to selectively quench the fluorescence of the undigested probe leaving the fluorescent signal of the fluorescently-labeled probe fragments. The fluorometric signals for miRNA-21 had a maximum excitation/emission wavelength of 488/518 nm. Most importantly, the biosensor was then applied for the accurate quantitative detection of miRNA-21 in exosome lysates extracted from human plasma and this method was able to successfully distinguish lung cancer patients from healthy people. This biosensor provides a simple, rapid, and a highly specific quantitative method for exosomal miRNA and has promising potential to be used in the early diagnosis of lung cancer.

Metabolomic analysis of plasma from normal-weight adults with hypo-HDL cholesterolemia by UPLC-QTOF MS.

High-density lipoprotein cholesterol (HDL-C) is negatively correlated with atherosclerotic cardiovascular disease. The prevalence of hypo-HDL cholesterolemia is as high as 33.9%. The plasma metabolomic differences between hypo-HDL cholesterolemia populations and normal controls were investigated using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Participants with hypo-HDL cholesterolemia and normal controls were clearly discriminated from each other on the orthogonal partial least squares-discriminant analysis score plot and a total of 90 differential metabolites were identified, including down-regulated phosphatidylserine [18:0/20:3(8Z,11Z,14Z)], phosphatidylcholine [19:0/18:3(6Z,9Z,12Z)], phosphatidylserine, phosphatidylethanolamine [18:0/20:4(5Z,8Z,11Z,13E) (15Ke)], etc., and up-regulated triglyceride [15:0/18:1(9Z)/18:3(9Z,12Z,15Z)][iso6], 13-methyl-1-tritriacontene, tridodecylamine, etc. Most of the changed metabolites were lipids, notably, a significant part of which were odd chain fatty acid incorporated lipids. Carnitine shuttle was the most significant metabolic pathway, except for the disturbed glycerophospholipid metabolism, glycosphingolipid metabolism and sphingolipid metabolism in participants with hypo-HDL cholesterolemia. We identified the key metabolites and metabolic pathways that may be changed in hypo-HDL cholesterolemia participants, providing useful clues for studying the metabolic mechanisms and for early prevention of hypo-HDL cholesterolemia and dyslipidemia.

Simultaneous Detection of VEGF and CEA by Time-Resolved Chemiluminescence Enzyme-Linked Aptamer Assay.

BACKGROUND: As two important tumor markers, vascular endothelial growth factor (VEGF) and carcinoembryonic antigen (CEA) have a great value for clinical application in the early diagnosis of cancer. Due to the complex composition of biological samples, the results from combined detection of CEA and VEGF are often taken as a comprehensive indicator in order to make an accurate judgment on a disease. However, most of the current methods can only be used to detect the content of one biomarker. Therefore, it is necessary to explore a simple, rapid, low-cost, and highly sensitive method for the simultaneous detection of CEA and VEGF. METHODS: Based on specific aptamers and magnetic separation, a time-resolved chemiluminescence enzyme-linked aptamer assay was developed for the simultaneous detections of CEA and VEGF in serum samples. RESULTS: Under the optimal conditions, the linear range of the calibration curve for VEGF was from 0.5 to 80 ng mL-1, and the limit of detection was 0.1 ng mL-1. The linear range of the calibration curve for CEA was 0.5 to 160 ng mL-1, and the limit of detection was 0.1 ng mL-1. The established method was applied to detect VEGF and CEA in serum samples. The results were consistent with those of commercial kits. CONCLUSION: The method has high sensitivity and can quickly obtain accurate results, which could greatly improve the measurement efficiency, reduce the cost, and also reduce the volume of sample consumed. It can be seen that the method established in this study has important application value and broad application prospect in clinical diagnosis.

Highly Sensitive Fluorescence-Linked Immunosorbent Assay for the Determination of Human IgG in Serum Using Quantum Dot Nanobeads and Magnetic Fe3O4 Nanospheres.

The aim of this study is to establish a new method with high sensitivity, accuracy, and stability for the determination of human IgG and then expand it to analyze severe acute respiratory syndrome corona virus 2 (SARS-CoV-2)-specific IgM and IgG, which is of great significance for the screening and diagnosis of COVID-19. In this study, the magnetic Fe3O4 nanospheres coupled with mouse antihuman IgG (Ab1IgG) were used as an immune capture probe (Fe3O4@Ab1IgG) to capture and separate the target, and rabbit antihuman IgG (Ab2IgG) coupled with highly luminescent quantum dot nanobeads (QBs) as a fluorescence detection probe (QBs@Ab2IgG) was used to realize high sensitivity detection. After the formation of a sandwich immunocomplex, the fluorescence intensity of the precipitate after magnetic separation was measured at the excitation wavelength of 370 nm. Under optimal conditions, a wide linear range varying from 0.005 to 40 ng·mL-1 was obtained for the detection of human IgG with a lower limit of detection at 4 pg·mL-1 (S/N = 3). The recoveries of intra- and interassays were 90.0-101.9 and 96.0-106.6%, respectively, and the relative standard deviations were 6.3-10.2 and 2.6-10.5%, respectively. Furthermore, the proposed method was successfully demonstrated to detect human IgG in serum samples, and the detection results were not statistically different (P > 0.05) from commercial enzyme-linked immunosorbent assay kits. This method is sensitive, fast, and accurate, which could be expanded to detect the specific IgM and IgG antibodies against SARS-CoV-2.

A novel fluorescent nanoprobe that based on poly(thymine) single strand DNA-templated copper nanocluster for the detection of hydrogen peroxide.

In this paper, a label-free fluorescence nanoprobe is constructed based on poly(thymine) single strand DNA-templated Copper nanocluster (denote as: T-CuNCs) for the detection of hydrogen peroxide. In the assay, the fluorescent T-CuNCs will generate though the reaction of Cu2+, poly(thymine) and sodium ascorbate. However, the hydroxyl radical (.OH) will generated in the presence of H2O2, which is able to induced the oxidative lesions of poly(thymine) single chain DNA and lead to the poly(thymine) being splitted into shorter or single oligonucleotide fragments and lose the ability to template the fluorescent T-CuNCs again. Therefore, H2O2 can be detected by monitoring the fluorescence strength change of T-CuNCs. The experimental results show that the fluorescence intensity change of T-CuNCs has fantastic linearity versus H2O2 concentration in the range of 1-30 µM (R2 = 0.9947) and 30-80 µM (R2 = 0.9972) with the limit of detection (LOD) as low as 0.5 µM (S/N = 3). More important, the fluorescent nanoprobe was also successfully utilized on the detection of H2O2 in serum samples. Therefore, a label-free, costless and effective fluorescence method has been established for the detection of H2O2, the intrinsic properties of the nanoprobe endow its more potential applications in chemical and biological study.

Fluorometric immunoassay for the simultaneous determination of the tumor markers carcinoembryonic antigen and cytokeratin 19 fragment using two kinds of CdSe/ZnS quantum dot nanobeads and magnetic beads.

A method is described for the simultaneous determination of the carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1). Two kinds of CdSe/ZnS quantum dot nanobeads (QBs), with emission maxima at 530 nm (green) and 585 nm (yellow), were used as labels, and magnetic beads (MBs) for separation. The MBs were used as substrates to couple CEA and CYFRA21-1 antibody for isolating the proteins. Then, the differently colored QBs were linked to the antibodies against CEA and CYFRA21-1, respectively. Following the formation of the immunocomplex, the intensities of the green and yellow emissions were measured at the same excitation wavelength of 340 nm. The detection limits are 0.1 ng⋅mL- 1 for CEA, and of 0.2 ng⋅mL- 1 for CYFRA21-1. The recoveries from spiked serum are 92.1 - 118.1% for CEA, and from 90.8% to 115.2% for CYFRA21-1, with the relative standard deviations of 6.3 - 12.3% and 7.1 - 11.8%. The method was successfully applied to the simultaneous determination of the two proteins in human serum sample (n = 45). The results correlated well with those of the chemiluminescent enzyme immunoassay kit. Graphical AbstractSchematic presentation of the fluorescence immunoassay for the simultaneous determination of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) based on quantum dot nanobeads (QBs) and magnetic beads (MBs) is reported. The intensities of two kinds of CdSe/ZnS QBs, with the emission maxima at 530 nm (green) and 585 nm (yellow) were measured at the same excitation wavelength of 340 nm.

A Novel Cell-Assisted Enhanced Chemiluminescence Strategy for Rapid and Label-Free Detection of Tumor Cells in Whole Blood.

In this study, an interesting phenomenon was found where cells (including tumor and normal cells) managed to significantly enhance chemiluminescence (CL) signals. The possible reaction mechanism may be that cells can be severely damaged by CL substrates, and the released contents, possibly proteins (such as cytochrome c), can remarkably magnify CL owing to the increased production of singlet oxygen. More importantly, based on the above phenomena, a novel cell-assisted enhanced CL strategy was proposed for the rapid and label-free detection of tumor cells. The complexes of aptamer sgc8c and streptavidin-modified magnetic beads were employed to recognize and isolate target tumor cells from whole blood. The enhanced CL intensity, which was triggered directly by the captured cells, was measured. The proposed strategy exhibited a good detection performance with a linear range from 200 to 10,000 cells/mL. The analysis can be finished in ∼30 min, and the limit of detection was down to 100 cells/mL. The recoveries and relative standard deviations were 97.81-102.71% and 3.46-12.71%, respectively. Moreover, the established method can successfully distinguish the leukemia patients from healthy people. Therefore, it provides a novel, rapid, and simple method for the determination of tumor cells, which can be used in further practice.

A highly sensitive colorimetric aptasensor for the detection of the vascular endothelial growth factor in human serum.

Early detection of cancer is of great significance for disease prevention and diagnosis. However, the levels of most cancer markers are quite low in the early stages of disease, so it is urgent to develop a highly sensitive detection method. In this study, a label-free and highly sensitive colorimetric strategy was developed for the detection of the vascular endothelial growth factor165 (VEGF165) in human serum. First, a convenient biosensor was constructed by immobilizing VEGF165 on a microplate, where aptamers bound with VEGF165 to form a complex. Then, streptavidin labeled-horseradish peroxidase (HRP-SA) combined with the complex via the interaction between streptavidin and biotin, thus catalyzing the 3,3',5,5'-tetramethylbenzidine (TMB) and H2O2 system to produce colored products. In the presence of target, immobilized VEGF165 and target competitively bound with the aptamers, resulting in a reduction of the colorimetric signal. Moreover, the optical density (OD) signal decreased with the increase of target concentration. The strategy showed a broad linear range (0.1-100 ng/mL) and a rather low detection limit of 10 pg/mL with good precision and selectivity. Further, the proposed method was successfully applied in detecting VEGF165 in human serum. The detection results of serum samples showed that the proposed assay had a high correlation with CLEIA kits (r = 0.971, P = 0.001). It has potential for application in clinical research and diagnosis.

Magnetic-assisted self-assembled aptamer/protein hybrid probes for efficient capture and rapid detection of cancer cells in whole blood.

Self-assembly of building blocks for constructing multifunctional materials has opened prospects for sensing applications in the biomedical fields. In particular, the combination of aptamer with DNA assembly-based nanotechnology has greatly improved the performance of cancer cell detection. Nevertheless, the cancer cell detection strategies of integrating aptamer with protein are relatively sparse. So we have developed a self-assembled aptamer method to realize the efficient capture and rapid detection of cancer cells by ingeniously combining aptamer modified magnetic nanoparticles as capture nanoprobes with self-assembled aptamer/protein hybrid probes (SAPPs) as signal amplification probes. By merely mixing the component materials together simultaneously, the SAPPs, integrating aptamer for cancer cell recognition with protein for amplifying signal, were fabricated by DNA-governed one-step assembly. In addition, the SAPPs-based method exhibits efficient capture, rapid (about 45 min) and specific CCRF-CEM detection performance, with limits of detection down to 75 cells/mL in buffer and 200 cells/mL in whole blood.

Simultaneous detection of carcinoembryonic antigen and neuron-specific enolase in human serum based on time-resolved chemiluminescence immunoassay.

In the clinical diagnosis of tumor, the immunological detection of single tumor markers may lead to errors and missed inspection. Therefore, it is necessary to establish an accurate and effective method for the simultaneous detection of multiple tumor markers. Thus, we developed a time-resolved chemiluminescence immunoassay (TRCLIA) to simultaneously detect carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in human serum. Horseradish peroxidase (HRP) and alkaline phosphatase (ALP) were used as the detection probes to label the monoclonal antibodies of CEA and NSE by strain-promoted azide-alkyne cycloaddition (SPAAC), respectively. Based on a sandwich immunoassay, the targets in the samples were captured by antibodies immobilized on the surface of carboxylate-modified polystyrene microspheres (CPSMS) and sandwiched by other antibodies labeled with HRP and ALP. Since HRP and ALP had different dynamic characteristics, the CEA and NSE signals were recorded at 0.5 s and 20 min, respectively, and cross-interference could be avoided effectively. The whole signal detection processes could be completed in 20 min. The linear ranges of CEA and NSE were 0.1-64 ng mL-1 and 0.05-64 ng mL-1 and the limits of detection were 0.085 ng mL-1 and 0.044 ng mL-1 (S/N = 2), respectively. Also, 45 human serum samples obtained from patients having lung disease were tested by TRCLIA and commercial chemiluminescence enzyme-linked immunoassay (CLEIA) kits with good correlation. The correlation coefficients of CEA and NSE were 0.985 and 0.970, respectively. The results demonstrated a novel, effective, reliable and convenient TRCLIA method for the clinical diagnosis of CEA and NSE. The TRCLIA method has the potential to be an effective clinical tool for the early screening of lung cancer and can be applied in clinical diagnosis.

Super-resolution imaging of cancer-associated carbohydrates using aptamer probes.

Globo H, as one of the most crucial cancer-associated carbohydrates, is exclusively overexpressed in a variety of cancers. However, the accurate localization and detailed morphology of globo H at the molecular level remain unclear. Here, we applied direct stochastic optical reconstruction microscopy (dSTORM) and relied on fluorophore-conjugated aptamers to solve the problem. The results showed that globo H organized as clusters on cell membranes with irregular shapes and different sizes from 100 to 300 nm. Significantly, globo H was found to have a higher expression level and larger clusters on various cancer cells than on non-cancer cells, which hinted that its specific distribution could be utilized for cancer diagnosis. Moreover, dual-color dSTORM imaging revealed the colocalization of globo H and other cancer-associated carbohydrates, and the clustering of globo H could be disrupted by the treatment of corresponding glycosidases, which indicated that these carbohydrates might intertwine in spatial organization and function cooperatively in cancers. Our work clarified the clustered distribution of globo H at the nanometer scale and revealed the potential interactions between cancer-associated carbohydrates, which paves the way for further understanding the relationship between the spatial structures and functions of carbohydrates in cancers.

The structural characteristics of mononuclear-macrophage membrane observed by atomic force microscopy.

Mononuclear macrophages are important immune cells in the organisms. The complicated membrane structure underlying the diverse functions of mononuclear-macrophage has been largely unresolved. As a representative of monocyte-derived macrophages, the membrane structure of PMA differentiated THP-1 cells was comprehensively investigated by AFM-based single molecule approaches. The rugged ectoplasmic side of mononuclear-macrophage membrane are significantly different from erythrocytes and mammalian somatic cell membranes. But the smooth lipid bilayer and the branched lipid raft domains obtained by proteinase K and MßCD treatment of the protein-covered cytoplasmic side, are common characteristics among all the studied cell membranes. This discovery of distinct organization of membrane proteins on both sides of mononuclear-macrophage membranes provides additional evidence for the asymmetry of membrane structure. The podosome-associated structures of mononuclear-macrophage were directly examined, and the independent localization of podosome domains and the lipid rafts was verified by in situ AFM, giving new insight into this multifunctional organelle.

Enhanced chemiluminescence enzyme-linked immunoassay for the determination of DNA methyltransferase 1 in human serum.

The occurrence of many diseases is closely related to the high expression of DNA methyltransferase 1 (DNMT1). However, most studies are focused on the detection of DNMT1 activity, a few are concerned with the detection of DNMT1 content. In this study, we developed a simple and highly sensitive chemiluminescence (CL) assay for the detection of DNMT1 content. In this method, anti-DNMT1 monoclonal antibody was coated on a polystyrene microplate to capture DNMT1. Then anti-DNMT1 polyclonal antibody and goat anti-rabbit immunoglobulin G with horseradish peroxidase (IgG-HRP) were respectively added to combine with captured DNMT1 to form a sandwich structure. Finally, the HRP could catalyze CL substrate and achieve CL signal response. Based on this novel sensitive strategy, the recovery percents were in the ranges from 71.5% to 91.0%. The precision of intra-assays and inter-assays were 5.45%-11.29% and 7.03%-11.25%, respectively. The method was successfully applied for the determination of DNMT1 in human serum. The detection results of serum samples showed that the proposed assay had a high correlation with enzyme-linked immunosorbent assay (ELISA) kit. Compared with the ELISA kit (limit of detection = 0.1 ng/mL), the method has a lower limit of detection of 0.042 ng/mL. Therefore, our method has the potential for the detection of DNMT1 content in clinical diagnosis.

High local coordination symmetry around the spin center and the alignment between magnetic and symmetric axes together play a crucial role in single-molecule magnet performance.

A tetradentate 8-hydroxyquinoline-based acyl hydrazone ligand (HL1 = 8-hydroxyquinoline-2-carboxaldehyde-(aminourea)hydrochloride) was elaborately used to construct a mononuclear dysprosium complex DyCl3HL1·CH3OH (1) with a nearly ideal pentagonal bipyramid coordination geometry (D5h) surrounding the Dy(iii) ion to achieve the significant performance of single-molecule magnets (SMMs). Meanwhile, the isolated high local symmetry center was successfully kept intact and further bridged to a series of double bipyramid systems by two phenolic oxygen atoms of the acyl hydrazone ligands (HL1 and HL2 = 8-hydroxyquinoline-2-carboxaldehyde-(benzoyl)hydrazine), with the formulae [Dy2Cl4(L1)2(CH3OH)2]·4C5H5N (2) and [Dy2Cl4(L2)2]·2CH3CN (3). In addition, the monodentate co-ligand anion was replaced by a larger sterically hindered ligand and a bidentate monovalent ß-diketonate anion to generate [Dy2(tfo)4(L2)2(EtOH)2] (4), [Dy2(tta)4(L2)2(EtOH)2]·2(EtOH) (5) and [Dy2(dbm)4(L2)2(EtOH)2] (6) (tfo = trifluoromethanesulfonic acid, dbm = dibenzoylmethane, tta = 4,4,4-trifluoro-1-(2-thienyl)-1,3-butanedione) with eight-coordinate geometry. Strikingly, the dynamic magnetic measurements revealed that complexes 1-3 did not display the expected significant SMM performance albeit they had high local symmetry. In combination with ab initio calculation, the alignment of the coordination symmetric axis and the magnetic easy axis dominates the molecular magnetic anisotropy, and the magnetic easy axis could be modulated by the distribution of coordination atoms with different electrostatic properties.