Split-aptamer mediated regenerable temperature-sensitive electrochemical biosensor for the detection of tumour exosomes.

In this paper, a split-aptamer mediated regenerable temperature-sensitive (SMRT) electrochemical biosensor was constructed for the detection of exosomes. The split-aptamer used in this SMRT biosensor was composed of two fragments, one of which was immobilized on the surface of an electrode via sulfhydryl groups and named split-a and the other was labelled with methylene blue and named split-b. The two fragments could form sandwich structures at the electrode surface via target-induced self-assembly in the presence of target exosomes at 4 °C in PBS, and then realizing the detection of exosomes via voltammetry. In addition, due to the temperature sensitivity of the split-aptamer, the electrode could be regenerated through temperature-induced disassembly of the sandwich structures. Consequently, the SMRT biosensor realized sensitive and specific analysis of target exosomes with a limit of detection of 1.5 × 106 particles/mL and could be quickly and easily regenerated by washing with PBS at 37 °C for 30 s without any additives. This is the first study on the construction of a reproducible electrochemical biosensor using a split-aptamer for the specific detection of tumour exosomes, and may provide an innovative strategy for the economical and efficient design of regenerable electrochemical biosensors.

Scutellarein relieves the death and inflammation of tubular epithelial cells in ischemic kidney injury by degradation of COX-2 protein.

OBJECTIVES: Acute kidney injury (AKI) is a clinical syndrome that usually caused by ischemia/reperfusion (I/R). Previous studies have revealed the protection of scutellarein against ischemia in nervous system. This study aimed to demonstrate the potential of scutellarein in ischemic AKI. METHODS: Animal model of ischemic AKI was established by clamping bilateral kidney pedicles in Sprague-Dawley rats. HK-2 cells were exposed to oxygen glucose deprivation/reoxygenation (OGD/R) to induce a cell model of AKI. The effects of scutellarein pre-treatment were detected by H&E staining, TUNEL, ELISA, CCK-8, LDH activity assay, ROS generation, flow cytometry, qRT-PCR and western blotting. Bioinformatic analysis was performed to probe the targets of scutellarein. RESULTS: The blood urea nitrogen (BUN) and serum creatinine (SCr) levels in rats treated with scutellarein were lower than that in model groups. Scutellarein suppressed the pathological injury of kidney, and dose-dependently inhibited the apoptosis and pro-inflammatory cytokines release (IL-1ß, IL-6 and IL-18). Scutellarein prevented OGD/R-induced HK-2 cell loss and cytotoxicity. ROS generation, apoptosis, and inflammation induced by OGD/R were all inhibited by scutellarein. By searching on the TCMSP and Symmap databases, COX-2 was screened out as a target of scutellarein. Scutellarein has no significant impacts on COX-2 mRNA expression, but could inhibit its protein level. Scutellarein promoted COX-2 protein degradation via enhancing autophagy. Furthermore, overexpression of COX-2 partly eliminated the renal protection of scutellarein in HK-2 cells. CONCLUSIONS: Scutellarein was suggested as a renal protective agent against ischemia-induced damage in AKI. The protective properties of scutellarein may be through inhibition of COX-2.

Large-scale translatome profiling annotates the functional genome and reveals the key role of genic 3' untranslated regions in translatomic variation in plants.

The translatome, a profile of the translational status of genetic information within cells, provides a new perspective on gene expression. Although many plant genomes have been sequenced, comprehensive translatomic annotations are not available for plants due to a lack of efficient translatome profiling techniques. Here, we developed a new technique termed 3' ribosome-profiling sequencing (3'Ribo-seq) for reliable, robust translatomic profiling. 3'Ribo-seq combines polysome profiling and 3' selection with a barcoding and pooling strategy. Systematic translatome profiling of different tissues of Arabidopsis, rice, and maize using conventional ribosome profiling (Ribo-seq) and 3'Ribo-seq revealed many novel translational genomic loci, thereby complementing functional genome annotation in plants. Using the low-cost, efficient 3'Ribo-seq technique and genome-wide association mapping of translatome expression (eGWAS), we performed a population-level dissection of the translatomes of 159 diverse maize inbred lines and identified 1,777 translational expression quantitative trait loci (eQTLs). Notably, local eQTLs are significantly enriched in the 3' untranslated regions of genes. Detailed eQTL analysis suggested that sequence variation around the polyadenylation (polyA) signal motif plays a key role in translatomic variation. Our study provides a comprehensive translatome annotation of plant functional genomes and introduces 3'Ribo-seq, which paves the way for deep translatomic analysis at the population level.

A novel molecularly imprinted sensor based on PtCu bimetallic nanoparticle deposited on PSS functionalized graphene with peroxidase-like activity for selective determination of puerarin.

In this work, PtCu bimetallic nanoparticle was deposited on poly (styrene sulfonate) (PSS) functionalized graphene (Gr) to form a nanocomposite PtCu/PSS-Gr and its enzyme-like activity was investigated. Benefiting from the synergistic effect from Pt and Cu monometal as well as the superior properties of PSS-Gr, such as large surface area, good dispersity, strong adsorption of substrate and additional peroxidase-like activity, the PtCu/PSS-Gr nanocomposite was demonstrated as an excellent peroxidase mimic to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. Combined with molecularly imprinted polymer (MIP), a new colorimetric approach for puerarin detection was proposed with the linear range of 2 × 10-5-6 × 10-4 mol L-1 and LOD of 1 × 10-5 mol L-1. The combination of MIP with PtCu/PSS-Gr nanocomposite not only endowed the determination of puerarin with high selectivity, but also realized the detection of small molecules which are neither substrate of the nanozyme nor substances with strong oxidizing or reducing activity by using peroxidase-like catalytic activity of nanozyme, expanding the application of nanozyme.

Flower-like MoS2 decorated with Cu2O nanoparticles for non-enzymatic amperometric sensing of glucose.

In this study, a novel nanohybrid consisting of flower-like MoS2 decorated with Cu2O nanoparticles has been successfully synthesized for non-enzymatic amperometric sensing of glucose. Structural characterizations revealed that Cu2O nanoparticles were highly dispersed on MoS2 nanosheets. Electrochemical performances were investigated by cyclic voltammetry (CV) and chronoamperometry. Compared to single Cu2O component, the-synthesized Cu2O/MoS2 nanohybrid showed superior electrocatalysis to the oxidation of glucose. The fabricated non-enzymatic amperometric glucose sensor exhibited a wide linear range from 0.01 to 4.0mM with a low detection limit of 1.0µM (S/N =3) and a high sensitivity of 3108.87µAmM-1cm-2. Meanwhile, the non-enzymatic sensor also possesses satisfactory stability, good reproducibility and high selectivity to interfering components of uric acid, dopamine and ascorbic acid. The excellent analytical performances are resulted from the synergistic effect provided by the Cu2O nanoparticals and MoS2 nanosheets.

De novo comparative transcriptome analysis provides new insights into sucrose induced somatic embryogenesis in camphor tree (Cinnamomum camphora L.).

BACKGROUND: Somatic embryogenesis is a notable illustration of cell totipotency, by which somatic cells undergo dedifferentiation and then differentiate into somatic embryos. Our previous work demonstrated that pretreatment of immature zygotic embryos with 0.5 M sucrose solution for 72 h efficiently induced somatic embryo initiation in camphor tree. To better understand the molecular basis of somatic embryogenesis induced by osmotic stress, de novo transcriptome sequencing of three tissues of camphor tree (immature zygotic embryos, sucrose-pretreated immature zygotic embryos, and somatic embryos induced from sucrose-pretreated zygotic embryos) were conducted using Illumina Hiseq 2000 platform. RESULTS: A total of 30.70 G high quality clean reads were obtained from cDNA libraries of the three samples. The overall de novo assembly of cDNA sequence data generated 205592 transcripts, with an average length of 998 bp. 114229 unigenes (55.56 % of all transcripts) with an average length of 680 bp were annotated with gene descriptions, gene ontology terms or metabolic pathways based on Blastx search against Nr, Nt, Swissprot, GO, COG/KOG, and KEGG databases. CEGMA software identified 237 out of 248 ultra-conserved core proteins as 'complete' in the transcriptome assembly, showing a completeness of 95.6 %. A total of 897 genes previously annotated to be potentially involved in somatic embryogenesis were identified. Comparative transcriptome analysis showed that a total of 3335 genes were differentially expressed in the three samples. The differentially expressed genes were divided into six groups based on K-means clustering. Expression level analysis of 52 somatic embryogenesis-related genes indicated a high correlation between RNA-seq and qRT-PCR data. Gene enrichment analysis showed significantly differential expression of genes responding to stress and stimulus. CONCLUSIONS: The present work reported a de novo transcriptome assembly and global analysis focused on gene expression changes during initiation and formation of somatic embryos in camphor tree. Differential expression of somatic embryogenesis-related genes indicates that sucrose induced somatic embryogenesis may share or partly share the mechanisms of somatic embryogenesis induced by plant hormones. This study provides comprehensive transcript information and gene expression data for camphor tree. It could also serve as an important platform resource for further functional studies in plant embryogenesis.