Zirconium-porphyrin-MOF-based oxidase-like nanozyme with oxygen vacancy for aflatoxin B1 colorimetric sensing.

In this study, a porous coordination network zirconium-porphyrin-based nanoparticle with oxygen vacancies (OVs) was prepared using acetic acid and benzoic acid as modulators via a simple hydrothermal method. The presence of OVs was confirmed by various characterization methods and was found to enhance oxygen uptake and activation. This resulted in the generation of more reactive peroxyl radicals (•O2 -) and led to an improved oxidase (OXD) mimetic activity. Additionally, it promoted 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) oxidation, with a low Km value of 0.07 mM and a high Vmax of 1.47 × 10-7 M·s-1. As aflatoxin B1 (AFB1) inhibits the Pt@PCN-222-ABTS nanozyme system, a colorimetric probe for AFB1 detection was constructed. The limit of detection (LOD) was 0.074 µg·L-1. This research presents a novel approach for designing a nanozymatic-based colorimetric method to analyze trace AFB1 residues in food.

Ultrasound and defect engineering-enhanced nanozyme with high laccase-like activity for oxidation and detection of phenolic compounds and adrenaline.

Perusing metal-based redox nanozyme offers new opportunity for pollutant removal and biosensor, but ultrasound (US)-driven laccase-like nanozyme remains a significant challenge, especially in combination with defect engineering strategy. Herein, the Cu2Ov@Ce-TCPP was synthesized by doping Ce3+ on the surface of Cu2O nanocube and then coating with the porphyrin sonosensitizer. The Ce-doped porphyrin metal-structure in nanozyme was demonstrated to generate oxygen vacancy defects, which could obviously promote the laccase-like activity of Cu2Ov@Ce-TCPP nanozyme under US. XPS characterization and density functional theory (DFT) theoretical calculation revealed that the ultrasonic stimulation is beneficial to accelerate the electron transfer rate and O2 adsorption to improve catalytic activity, and Cu2Ov@Ce-TCPP nanozyme exhibits low adsorption energy and activation energy due to the presence of oxygen defect site, resulting in high laccase-like activity. The interaction between Ce atom and porphyrin structure also improved the sonocatalytic ability of the nanozyme. Meanwhile, Cu2Ov@Ce-TCPP nanozyme has been used for detecting and degrading a series of phenolic compounds. The detection adrenaline method has a linear range of 3.3-1000 µM and a detection limit as low as 0.96 µM with good reproducibility. The developed US-enhancing and recyclable laccase-like nanozyme system provides a promising strategy for the oxidation and detection of phenolic compounds.

SERS detection of Hg2+ and aflatoxin B1 through on-off strategy of oxidase-like Au@HgNPs/carbon dots.

In this work, cerium-doped carbon dots (Ce-CDs) both as a reducing agent and template hybrid gold nanoparticles (AuNPs) with weak oxidase-like (OXD) activity was synthesized for the detection of Hg2+ and aflatoxin B1 (AFB1). The AuNPs can catalyze efficiently mercury ion (Hg2+) reduction to the metallic (Hg0) to form Au-Hg amalgam (Au@HgNPs). The obtained Au@HgNPs with strong OXD-like activity oxidize without Raman-active leucomalachite green (LMG) into the Raman-active malachite green (MG) and simultaneously as the SERS substrates by the formed Raman "hot spot" through MG-induced Au@HgNPs aggregation. While AFB1 was introduced resulting in the SERS intensity decreasing due to Hg2+ with AFB1 via carbonyl group to inhibit the aggregation of Au@HgNPs. The work paves a new path for the design of a nanozyme-based SERS protocol for tracing Hg2+ and AFB1 residues in foodstuff analysis.

Ganoderma Lucidum Polysaccharides Enhance the Abscopal Effect of Photothermal Therapy in Hepatoma-Bearing Mice Through Immunomodulatory, Anti-Proliferative, Pro-Apoptotic and Anti-Angiogenic.

Hepatocellular carcinoma is a malignant tumor with high morbidity and mortality, a highly effective treatment with low side effects and tolerance is needed. Photothermal immunotherapy is a promising treatment combining photothermal therapy (PTT) and immunotherapy. PTT induces the release of tumor-associated antigens by ablating tumor and Ganoderma lucidum polysaccharides (GLP) enhance the antitumor immunity. Results showed that Indocyanine Green (ICG) was successfully encapsulated into SF-Gel. ICG could convert light to heat and SF-Gel accelerates the photothermal effect in vitro and in vivo. PTT based on ICG/ICG-SF-Gel inhibited the growth of primary and distal tumors, GLP enhanced the inhibitory efficacy. ICG/ICG-SF-Gel-based PTT and GLP immunotherapy improved the survival time. ICG/ICG-SF-Gel-based PTT induces tumor necrosis and GLP enhanced the photothermal efficacy. ICG/ICG-SF-Gel-based PTT inhibited cell proliferation and angiogenesis, induced cell apoptosis, enhanced cellular immunity, and GLP enhanced these effects. In conclusion, GLP could enhance the abscopal effect of PTT in Hepatoma-bearing mice.

Intracerebroventricular Delivery of Recombinant NAMPT Deters Inflammation and Protects Against Cerebral Ischemia.

Our previous study indicated that nicotinamide phosphoribosyltransferase (NAMPT) is released from cells and might be an important extracellular neuroprotective factor in brain ischemia. Here, we tested whether NAMPT protects against ischemic brain injury when administered directly into the intracerebroventricular (ICV) compartment of the cranium. Recombinant NAMPT protein (2 µg) was delivered ICV in mice subjected to 45-min middle cerebral artery occlusion (MCAO), and the effects on infarct volume, sensorimotor function, microglia/macrophage polarization, neutrophil infiltration, and BBB integrity were analyzed. The results indicate that ICV administration of NAMPT significantly reduced infarct volume, retained its beneficial properties even when ICV administration was delayed by 6 h after MCAO, and improved neurological outcomes. NAMPT treatment inhibited pro-inflammatory microglia/macrophages, promoted microglia/macrophage polarization toward the anti-inflammatory phenotype, and reduced the infiltration of neutrophils into the perilesional area after brain ischemia. In vitro studies indicated that multiple pro-inflammatory microglial markers/cytokines were downregulated while multiple anti-inflammatory microglial markers/cytokines were induced in primary microglial cultures treated with NAMPT protein. NAMPT treatment also fortified the blood-brain barrier (BBB), as shown by reduced extravascular leakage of the small-molecule tracer Alexa Fluor 555 Cadaverine and larger-sized endogenous IgGs into brain parenchyma. Thus, NAMPT may protect against ischemic brain injury partly through a novel anti-inflammatory mechanism, which in turn maintains BBB integrity and reduces the infiltration of peripheral inflammatory cells. Taken together, these results provide validation of recombinant NAMPT delivery into the extracellular space as a potential neuroprotective strategy for stroke.

Regulation of Caveolin-1 and Junction Proteins by bFGF Contributes to the Integrity of Blood-Spinal Cord Barrier and Functional Recovery.

The blood-spinal cord barrier (BSCB) plays important roles in the recovery of spinal cord injury (SCI), and caveolin-1 is essential for the integrity and permeability of barriers. Basic fibroblast growth factor (bFGF) is an important neuroprotective protein and contributes to the survival of neuronal cells. This study was designed to investigate whether bFGF is beneficial for the maintenance of junction proteins and the integrity of the BSCB to identify the relations with caveolin-1 regulation. We examined the integrity of the BSCB with Evans blue dye and fluorescein isothiocyanate-dextran extravasation, measured the junction proteins and matrix metalloproteinases, and evaluated the locomotor function recovery. Our data indicated that bFGF treatment improved the recovery of BSCB and functional locomotion in contusive SCI model rats, reduced the expression and activation of matrix metalloproteinase-9, and increased the expressions of caveolin-1 and junction proteins, including occludin, claudin-5, p120-catenin, and ß-catenin. In the brain, in microvascular endothelial cells, bFGF treatment increased the levels of junction proteins, caveolin-1 small interfering RNA abolished the protective effect of bFGF under oxygen-glucose deprivation conditions, and the expression of fibroblast growth factor receptor 1 and co-localization with caveolin-1 decreased significantly, which could not be reversed by bFGF treatment. These findings provide a novel mechanism underlying the beneficial effects of bFGF on the BSCB and recovery of SCI, especially the regulation of caveolin-1.

Retinoic Acid Prevents Disruption of Blood-Spinal Cord Barrier by Inducing Autophagic Flux After Spinal Cord Injury.

Spinal cord injury (SCI) induces the disruption of the blood-spinal cord barrier (BSCB), which leads to infiltration of blood cells, inflammatory responses and neuronal cell death, with subsequent development of spinal cord secondary damage. Recent reports pointed to an important role of retinoic acid (RA), the active metabolite of the vitamin A, in the induction of the blood-brain barrier (BBB) during human and mouse development, however, it is unknown whether RA plays a role in maintaining BSCB integrity under the pathological conditions such as SCI. In this study, we investigated the BSCB protective role of RA both in vivo and in vitro and demonstrated that autophagy are involved in the BSCB protective effect of RA. Our data show that RA attenuated BSCB permeability and also attenuated the loss of tight junction molecules such as P120, ß-catenin, Occludin and Claudin5 after injury in vivo as well as in brain microvascular endothelial cells. In addition, RA administration improved functional recovery of the rat model of trauma. We also found that RA could significantly increase the expression of LC3-II and decrease the expression of p62 both in vivo and in vitro. Furthermore, combining RA with the autophagy inhibitor chloroquine (CQ) partially abolished its protective effect on the BSCB and exacerbated the loss of tight junctions. Together, our studies indicate that RA improved functional recovery in part by the prevention of BSCB disruption via the activation of autophagic flux after SCI.

bFGF Protects Against Blood-Brain Barrier Damage Through Junction Protein Regulation via PI3K-Akt-Rac1 Pathway Following Traumatic Brain Injury.

Many traumatic brain injury (TBI) survivors sustain neurological disability and cognitive impairments due to the lack of defined therapies to reduce TBI-induced blood-brain barrier (BBB) breakdown. Exogenous basic fibroblast growth factor (bFGF) has been shown to have neuroprotective function in brain injury. The present study therefore investigates the beneficial effects of bFGF on the BBB after TBI and the underlying mechanisms. In this study, we demonstrate that bFGF reduces neurofunctional deficits and preserves BBB integrity in a mouse model of TBI. bFGF suppresses RhoA and upregulates tight junction proteins, thereby mitigating BBB breakdown. In vitro, bFGF exerts a protective effect on BBB by upregulating tight junction proteins claudin-5, occludin, zonula occludens-1, p120-catenin, and ß-catenin under oxygen glucose deprivation/reoxygenation (OGD) in human brain microvascular endothelial cells (HBMECs). Both the in vivo and in vitro effects are related to the activation of the downstream signaling pathway, PI3K/Akt/Rac-1. Inhibition of the PI3K/Akt or Rac-1 by specific inhibitors LY294002 or si-Rac-1, respectively, partially reduces the protective effect of bFGF on BBB integrity. Overall, our results indicate that the protective role of bFGF on BBB involves the regulation of tight junction proteins and RhoA in the TBI model and OGD-induced HBMECs injury, and that activation of the PI3K/Akt /Rac-1 signaling pathway underlies these effects.

FGFR4 and TGF-ß1 expression in hepatocellular carcinoma: correlation with clinicopathological features and prognosis.

OBJECTIVE: To investigate the expression and correlation of transforming growth factor-ß1 (TGF-ß1) and fibroblast growth factor receptor 4 (FGFR4) in human hepatocellular carcinoma (HCC) and the relationship with clinicopathological features and prognosis. MATERIALS AND METHODS: The expression of TGF-ß1 and FGFR4 in 126 HCC samples was detected immunohistochemically. Combined with clinical postoperative follow-up data, the expression of TGF-ß1 and FGFR4 in HCC and the relationship with the prognosis of patients were analyzed by statistically. RESULTS: The positive expression rate of TGF-ß1 was 84.1% (106/126) in tumors, and that in peritumoral liver tissues was 64.3% (81/126); the positive expression rate of FGFR4 in tumors was 74.6% (94/126) and that in peritumoral liver tissues was 57.1% (72/126). The expression of TGF-ß1 and FGFR4 in the carcinoma tissues was significantly higher than that in peritumoral liver tissues (p < 0.05). Intratumoral TGF-ß1 and FGFR4 expression was associated with TNM stage (p < 0.05). TGF-ß1 and FGFR4 expression levels didn't significantly correlate with other clinicopathological parameters, including age, sex, tumor size, serum AFP level, tumor differentiation, lymph node metastasis, etc. (p > 0.05). TGF-ß1 expression was positively correlated with FGFR4 expression (r = 0.595, p < 0.05). Patients with positive FGFR4 or TGF-ß1 expression had shorter overall survival compared with negative expression (p < 0.05). CONCLUSIONS: The expression of TGF-ß1 and FGFR4 could make synergy on the occurrence and progression of HCC, and may be used as prognosis indicators for HCC patients.

Spectrofluorimetric determination of fluoroquinolones in honey with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone in the presence of ß-cyclodextrin.

A simple and sensitive spectrofluorimetric method was developed for the determination of four fluoroquinolone antibacterials namely norfloxacin (NOR), ofloxacin (OFL), ciprofloxacin (CIP) and gatifloxacin (GAT) in honey through charge transfer (CT) complex formation with 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), and then the inclusion complexes of FQs-DDQ with ß-cyclodextrin (ß-CD) were formed, which resulted in drastic fluorescence enhancement. The effect of several parameters including the concentration of reactants, reaction temperature, time and ultrasonic treatment on the efficiency of the proposed method involving CT reaction and inclusion interaction was systematically investigated. Under the optimum conditions, the limits of detection (LODs) for four FQs in honey varied from 11.6 to 15.4 µg/kg (signal-to-noise ratio (S/N) = 3). The intra- and interday relative standard deviations (RSDs) were 1.6-4.0 % (n = 5) for four FQs. The calibration graph was linear from 42.8 to 1346.8 µg/kg with correlation coefficients not less than 0.9905. The recoveries of four FQs at three different spiked concentrations in honey samples ranged from 80.9 % to 92.8 %. The results indicated that the method was successfully applied for analyzing FQs in honey.

Aluminium sensitized spectrofluorimetric determination of fluoroquinolones in milk samples coupled with salting-out assisted liquid-liquid ultrasonic extraction.

An aluminium sensitized spectrofluorimetric method coupled with salting-out assisted liquid-liquid ultrasonic extraction for the determination of four widely used fluoroquinolones (FQs) namely norfloxacin (NOR), ofloxacin (OFL), ciprofloxacin (CIP) and gatifloxacin (GAT) in bovine raw milk was described. The analytical procedure involves the fluorescence sensitization of aluminium (Al(3+)) by complexation with FQs, salting-out assisted liquid-liquid ultrasonic extraction (SALLUE), followed by spectrofluorometry. The influence of several parameters on the extraction (the salt species, the amount of salt, pH, temperature and phase volume ratio) was investigated. Under optimized experimental conditions, the detection limits of the method in milk varied from 0.009 µg/mL for NOR to 0.016 µg/mL for GAT (signal-to-noise ratio (S/N)=3). The relative standard deviations (RSD) values were found to be relatively low (0.54-2.48% for four compounds). The calibration graph was linear from 0.015 to 2.25 µg/mL with coefficient of determinations not less than 0.9974. The methodology developed was applied to the determination of FQs in bovine raw milk samples. The main advantage of this method is simple, accurate and green. The method showed promising applications for analyzing polar analytes especially polar drugs in various sample matrices.

Cloud-point extraction and reversed-phase high-performance liquid chromatography for the determination of synthetic phenolic antioxidants in edible oils.

A cloud-point extraction (CPE) method using Triton X-114 (TX-114) nonionic surfactant was developed for the extraction and preconcentration of propyl gallate (PG), tertiary butyl hydroquinone (TBHQ), butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) from edible oils. The optimum conditions of CPE were 2.5% (v/v) TX-114, 0.5% (w/v) NaCl and 40 min equilibration time at 50 °C. The surfactant-rich phase was then analyzed by reversed-phase high-performance liquid chromatography with ultraviolet detection at 280 nm, using a gradient mobile phase consisting of methanol and 1.5% (v/v) acetic acid. Under the studied conditions, 4 synthetic phenolic antioxidants (SPAs) were successfully separated within 24 min. The limits of detection (LOD) were 1.9 ng mL(-1) for PG, 11 ng mL(-1) for TBHQ, 2.3 ng mL(-1) for BHA, and 5.9 ng mL(-1) for BHT. Recoveries of the SPAs spiked into edible oil were in the range 81% to 88%. The CPE method was shown to be potentially useful for the preconcentration of the target analytes, with a preconcentration factor of 14. Moreover, the method is simple, has high sensitivity, consumes much less solvent than traditional methods, and is environment-friendly. Practical Application: The method established in this article uses less organic solvent to extract SPAs from edible oils; it is simple, highly sensitive and results in no pollution to the environment.