Choline Phosphate-Grafted Nanozymes as Universal Extracellular Vesicle Probes for Bladder Cancer Detection.

Urinary extracellular vesicles (uEVs) are regarded as highly promising liquid-biopsy biomarkers for the early diagnosis and prognosis of bladder cancer (BC). However, detection of uEVs remains technically challenging owing to their huge heterogeneity and ultralow abundance in real samples. We herein present a choline phosphate-grafted platinum nanozyme (Pt@CP) that acts as a universal EV probe for the construction of a high-throughput and high-sensitivity immunoassay, which allowed multiplex profiling of uEV protein markers for BC detection. With the Pt@CP-based immunoassays, three uEV protein markers (MUC-1, CCDC25, and GLUT1) were identified for BC, by which the BC cases (n = 48), cystitis patients (n = 27), and healthy donors (n = 24) were discriminated with high clinical sensitivity and specificity (area under curve = 98.3%). For the BC cases (n = 9) after surgery, the Pt@CP-based immunoassay could report the postoperative residual tumor that cannot be observed by cystoscopy, which is clinically significant for assessing BC recurrence. This work provides generally high sensitivity for EV detection, facilitating the discovery and clinical use of EV-based biomarkers.

In Vivo Fluorescent Labeling of Foam Cell-Derived Extracellular Vesicles as Circulating Biomarkers for In Vitro Detection of Atherosclerosis.

Real-time monitoring of the development of atherosclerosis (AS) is key to the management of cardiovascular disease (CVD). However, existing laboratory approaches lack sensitivity and specificity, mostly due to the dearth of reliable AS biomarkers. Herein, we developed an in vivo fluorescent labeling strategy that allows specific staining of the foam cell-derived extracellular vesicles (EVs) in atherosclerotic plaques, which are released into the blood as circulating biomarkers for in vitro detection of AS. This strategy relies on a self-assembled nanoprobe that could recognize foam cells specifically, where the probe is degraded by the intracellular HClO to produce a trifluoromethyl-bearing boron-dipyrromethene fluorophore (termed B-CF3), a lipophilic dye that can be transferred to the exosomal membranes. These circulating B-CF3-stained EVs can be detected directly on a fluorescence spectrometer or microplate reader without resorting to any sophisticated analytical method. This liquid-biopsy format enables early detection and real-time differentiation of lesion vulnerability during AS progression, facilitating effective CVD management.

Modulation of the binding ability to biomacromolecule, cytotoxicity and cellular imaging property for ionic liquid mediated carbon dots.

For the preparation of carbon dots (CDs), a variety of carbon sources and synthetic protocols are available which endow CDs with variable and unpredictable properties. In the present study, three CDs were developed with ionic liquid 1-butyl-3-methylimidazolium dicyanamide as the precursor through ethanol-thermal and hydrothermal strategies, termed as E-CDs and H-CDs, respectively. The features of these carbon dots, i.e., their physicochemical and optical properties, their interactions with bovine serum albumin (BSA) as well as their imaging capability were investigated with respect to the CDs prepared with microwave assisted approach (W-CDs). E-CDs and H-CDs were demonstrated to exhibit similar framework structures and optical properties, and they exhibited larger particle-sizes than that of W-CDs. In addition, the increase of ethanol-thermal and hydrothermal reaction time strengthened the quantum yields of the CDs and promoted their binding capability with BSA. E-CDs and H-CDs showed similar cytotoxicity on normal (LX-2) and cancer (SK-Hep-1) cells. We further found that these CDs may readily enter the cells within 5 min, while the fluorescence of hydrophilic E-CDs and H-CDs was very weak with respect to that of hydrophobic W-CDs in cell imaging. On the other hand, all the CDs exhibited little impact on the level of intracellular reactive oxygen species. The present study is conducive to guide the preparation of suitable carbon dots for different application scenarios.

miR-124-3p Inhibits Microglial Secondary Inflammation After Basal Ganglia Hemorrhage by Targeting TRAF6 and Repressing the Activation of NLRP3 Inflammasome.

Objectives: Intracerebral hemorrhage (ICH) represents a serious central nervous system emergency with high morbidity and mortality, and the basal ganglia is the most commonly affected brain region. Differentially expressed microRNAs (miRs) have recently been highlighted to serve as potential diagnostic biomarkers and therapeutic targets for ICH. This study investigated the mechanism of miR-124-3p in microglial secondary inflammation after ICH. Methods: In this study, 48 patients with primary basal ganglia ICH and 48 healthy volunteers were selected and venous blood was collected from all patients on the second morning of admission (within 24 h of stroke onset). The expression of miR-124-3p in serum was detected by RT-qPCR. Three months after ICH, the patients were assessed by modified Rankin Scale (mRS), and the correlation between miR-124-3p expression and mRS score was analyzed by Pearson. The inflammatory response of microglia was induced by lipopolysaccharide (LPS) to establish the cell model of microglial inflammation. miR-124-3p expression patterns were detected in the serum of ICH patients and healthy volunteers, normal microglia, and LPS-induced microglia. The miR-124-3p mimic was transfected into LPS-induced microglia, followed by measurement of the inflammatory factors, apoptosis rate, and cell viability. The target gene of miR-124-3p was predicted and verified. The expression patterns of tumor necrosis factor receptor-associated factor 6 (TRAF6) were detected. pcDNA3.1 and pcDNA3.1-TRAF6 were transfected into LPS-induced HMC3 cells, and nucleotide-binding oligomerization domain-like receptor (NLR) pyrin domain-containing 3 (NLRP3) expression patterns were determined. Lastly, the effects of TRAF6 overexpression on apoptosis, cell viability, and inflammation in HMC3 cells were measured. Results: miR-124-3p was downregulated in the serum of basal ganglia ICH patients and LPS-induced microglia, and miR-124-3p expression was negatively correlated with mRS. Overexpression of miR-124-3p reduced the inflammatory factors and apoptosis rate and promoted cell activity in LPS-induced microglia. miR-124-3p was found to target TRAF6. Overexpression of TRAF6 enhanced the expression of NLRP3 inflammasome, inflammatory factors and apoptosis rate, and reduced cell viability. Conclusion: Our findings indicate that miR-124-3p repressed the activation of NLRP3 inflammasome by targeting TRAF6, thus inhibiting microglial secondary inflammation after ICH in basal ganglia.

Identification and functional analysis of differentially expressed genes associated with cerebral ischemia/reperfusion injury through bioinformatics methods.

Cerebral ischemia/reperfusion (I/R) injury results in detrimental complications. However, little is known about the underlying molecular mechanisms involved in the reperfusion stage. The aim of the present study was to identify a gene expression profile associated with cerebral ischemia/reperfusion injury. The GSE23160 dataset, which comprised data from sham control samples and post­I/R injury brain tissues that were obtained using a middle cerebral artery occlusion (MCAO) model at 2, 8 and 24 h post­reperfusion, was downloaded from the Gene Expression Omnibus database. The differentially expressed genes (DEGs) in the MCAO samples compared with controls were screened using the GEO2R web tool. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for DEGs was performed using the online tool DAVID. Furthermore, a protein­protein interaction (PPI) network was constructed using the STRING database and Cytoscape software. In total, 32 DEGs at 2 h post­reperfusion, 39 DEGs at 8 h post­reperfusion and 91 DEGs at 24 h post­reperfusion were identified, while 15 DEGs were common among all three groups. GO analysis revealed that the DEGs at all three time­points were enriched in 'chemotaxis' and 'inflammatory response' terms, while KEGG pathway analysis demonstrated that DEGs were significantly enriched in the 'chemokine signaling pathway'. Furthermore, following PPI network construction, Cxcl1 was identified as the only hub gene that was common among all three time­points. In conclusion, the present study has demonstrated a global view of the potential molecular differences following cerebral I/R injury and may contribute to an improved understanding of the reperfusion stage, which may ultimately aid in the development of future clinical strategies.

Highly efficient removal of chromium(VI) from aqueous solution using polyaniline/sepiolite nanofibers.

Polyaniline/sepiolite (PANI/sepiolite) nanofibers were prepared by in situ chemical oxidation polymerization in the presence of sepiolite. The effect of aniline/sepiolite weight ratio on the nanostructure of PANI/sepiolite composites was investigated by field-emission scanning electron microscopy. The adsorption of Cr(VI) onto the PANI/sepiolite nanofibers was highly dependent on pH values. The pseudo-second-order and Langmuir isothermal models can well describe the adsorption kinetics and adsorption isotherm, respectively. The maximum adsorption capacity of the PANI/sepiolite nanofibers for Cr(VI) was up to 206.6 mg/g at 25 °C and increased with the increase in temperature. Desorption experiments indicated that PANI/sepiolite can be regenerated and reused for two consecutive cycles with no loss of its removal efficiency. PANI/sepiolite nanofibers can be used as a highly efficient and economically viable adsorbent for Cr(VI) removal due to their excellent adsorption characteristics.

Removal of hexavalent chromium from aqueous solution using exfoliated polyaniline/montmorillonite composite.

Exfoliated polyaniline/montmorillonite (PANI/MMT) composites with nanosheet structure were successfully prepared by in situ chemical oxidation polymerization with MMT platelets as the scaffold. Amphoteric polymer, (2-methacryloyloxyethyl)trimethyl ammonium chloride and methacrylate acid copolymer, was used to modify montmorillonite and a large number of carboxylic acids were introduced on the surface of the clay platelets, which can be used as a dopant of PANI and play a 'bridge' role to combine PANI with clay. Adsorption experiments were carried out to study the effects of pH, contact time, Cr(VI) concentration, adsorbent dose and temperature. The adsorption of Cr(VI) on the PANI/MMT was highly pH dependent and the adsorption kinetics followed a pseudo-second-order model. The Langmuir isothermal model described the adsorption isotherm data well and the maximum adsorption capacity increased with the increase in temperature. Thermodynamic investigation indicated that the adsorption process is spontaneous, endothermic and marked with an increase in randomness at the adsorbent - liquid interface. The maximum adsorption capacity of the PANI/MMT composites for Cr(VI) was 308.6 mg/g at 25 °C. The excellent adsorption characteristic of exfoliated PANI/MMT composites will render it a highly efficient and economically viable adsorbent for Cr(VI) removal.

[Spectral analysis of cyanobacteria chlorophyll in polluted water].

The polluted water with abundant nourishment cause phytoplankton, such as cyanobacteria, to grow rapidly, which brings great harm to environment. In the present paper, the absorption spectrum of cyanobacteria was measured and analyzed in order to estimate the content of the chlorophyll accurately. The same amount of cyanobacteria was separately cultured in pure water and lake water for different time. The chlorophyll was extracted from the cyanobacteria for the same time by 95% of ethanol. Then the ethanol extract was tested by ultraviolet visible spectrometry. The results show that the absorption spectrum of the chlorophyll has three absorption peaks at 279.5, 436.0 and 664.5 nm respectively. However, the absorbency at 279.5 nm cannot reflect the content of the chlorophyll. The absorbencies at 436.0 and 664.5 nm have linear relationship with the content of chlorophyll. Moreover, the dispersion between the absorbency at 436.0 nm and the absorbency at 664. 5 nm can reflect the content of chlorophyll more accurately. The research provides the experimental and theoretical basis for the highly accurate detection of the water quality.