Huzhi Zheng

Study on the fluorescence characteristics of carbon dots.

Herein, we prepared water-soluble fluorescent carbon dots with diameter about 1.5 nm from cheap commercial lampblack. These fluorescent carbon nanoparticles are stable toward photobleaching and stable in water for more than half a year without fluorescence decrease. In order to improve its fluorescence properties, we passivated these nanoparticles with bisamino-terminated polyethylene glycol (PEG(1500 N)). Therefore, both fluorescence quantum yield and lifetime increased after this progress. In addition, the passivated carbon dots were more inert to solvent than the bare one and showed different responses to pH change.

Copper nanoclusters as a highly sensitive and selective fluorescence sensor for ferric ions in serum and living cells by imaging

A simple, one-step facile route for preparation of water soluble and fluorescent Cu nanoclusters (NCs) stabilized by tannic acid (TA) is described. The as-prepared TA capped Cu NCs (TA-Cu NCs) are characterized by UV-vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, luminescence, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The TA-Cu NCs show luminescence properties having excitation and emission maxima at 360 nm and 430 nm, respectively, with a quantum yield of about 14%. The TA-Cu NCs are very stable even in 0.3 M NaCl, and their luminescent properties show pH independent. The fluorescence (FL) of Cu NCs is strongly quenched by Fe(3+) through an electron transfer mechanism, but not by other metal ions. Furthermore, the FL of the TA-Cu NCs shows no changes with the addition of Fe(2+) or H2O2 individually. On this basis, a facile chemosensor was developed for rapid, reliable, sensitive, and selective sensing of Fe(3+) ions with detection limit as low as 10 nM and a dynamic range from 10 nM to 10 μM. The proposed sensor was successfully used for the determination of iron contents in serum samples. Importantly, the Cu NCs-based FL probe showed long-term stability, good biocompatibility and very low cytotoxicity. It was successfully used for imaging ferric ions in living cells, suggesting the potential application of Cu NCs fluorescent probe in clinical analysis and cell imaging.

CdTe quantum dots as a highly selective probe for prion protein detection: colorimetric qualitative, semi-quantitative and quantitative detection.

CdTe quantum dots (QDs) were used as a highly selective probe for the detection of prion protein. Orange-emitting precipitates appeared within 30s of the addition of recombination prion protein (rPrP) to a solution of green-emitting CdTe QDs. This allowed colorimetric qualitative and semi-quantitative detection of rPrP. The decrease in fluorescence intensity of the supernatant could be used for quantitative detection of rPrP. The fluorescence intensity of the supernatant was inversely proportional to the rPrP concentration from 8 to 200 nmol L(-1) (R(2)=0.9897). Transmission electron microscopy results showed that fibrils existed in the precipitates and these were partly transformed to amyloid plaques after the addition of rPrP.

An aptamer-functionalized gold nanoparticle biosensor for the detection of prion protein

Prions are a special class of pathogens that cause a number of fatal neurodegenerative diseases in mammals. This paper presents a very simple and convenient biosensor for detecting prion protein, in which a prion protein aptamer was used as the molecular recognition group and gold nanoparticles were used as the signal report group. Binding of the target molecular prion protein resulted in the enhancement of resonance light scattering (RLS) by the gold nanoparticles. A linear relationship was then identified between the enhanced RLS intensity and the concentration of prion protein in the range 0.2 to 50 nmol L−1, with a limit of detection of 0.07 nmol L−1 (3σ). The biosensor has very good selectivity for prion protein without interference by coexisting proteins, amino acids or metal ions. This “aptasensor” offers a rapid, selective and sensitive route for prion protein detection and has good potential for use in practical applications.

Efficacy of NGR peptide-modified PEGylated quantum dots for crossing the blood–brain barrier and targeted fluorescence imaging of glioma and tumor vasculature

Delivery of imaging agents to brain glioma is challenging because the blood-brain barrier (BBB) functions as a physiological checkpoint guarding the central nervous system from circulating large molecules. Moreover, the ability of existing probes to target glioma has been insufficient and needs to be improved. In present study, PEG-based long circulation, CdSe/ZnS quantum dots (QDs)-based nanoscale and fluorescence, asparagines-glycine-arginine peptides (NGR)-based specific CD13 recognition were integrated to design and synthesize a novel nanoprobe by conjugating biotinylated NGR peptides to avidin-PEG-coated QDs. Our data showed that the NGR-PEG-QDs were nanoscale with less than 100 nm and were stable in various pH (4.0~8.0). These nanomaterials with non-toxic concentrations could cross the BBB and target CD13-overexpressing glioma and tumor vasculature in vitro and in vivo, contributing to fluorescence imaging of this brain malignancy. These achievements allowed groundbreaking technological advances in targeted fluorescence imaging for the diagnosis and surgical removal of glioma, facilitating potential transformation toward clinical nanomedicine.

Aptamer-conjugated PEGylated quantum dots targeting epidermal growth factor receptor variant III for fluorescence imaging of glioma

The extent of resection is a significant prognostic factor in glioma patients. However, the maximum safe resection level is difficult to determine due to the inherent infiltrative character of tumors. Recently, fluorescence-guided surgery has emerged as a new technique that allows safe resection of glioma. In this study, we constructed a new kind of quantum dot (QD)-labeled aptamer (QD-Apt) nanoprobe by conjugating aptamer 32 (A32) to the QDs surface, which can specially bind to the tumors. A32 is a single-stranded DNA capable of binding to the epidermal growth factor receptor variant III (EGFRvIII) specially distributed on the surface of glioma cells. To detect the expression of EGFRvIII in human brain tissues, 120 specimens, including 110 glioma tissues and 10 normal brain tissues, were examined by immunohistochemistry, and the results showed that the rate of positive expression of EGFRvIII in the glioma tissues was 41.82%, and 0.00% in normal brain tissues. Besides, the physiochemical properties of QD-Apt nanoparticles (NPs) were thoroughly characterized. Biocompatibility of the NPs was evaluated, and the results suggested that the QD-Apt was nontoxic in vivo and vitro. Furthermore, the use of the QD-Apt in labeling glioma cell lines and human brain glioma tissues, and target gliomas in situ was also investigated. We found that not only could QD-Apt specially bind to the U87-EGFRvIII glioma cells but also bind to human glioma tissues in vitro. Fluorescence imaging in vivo with orthotopic glioma model mice bearing U87-EGFRvIII showed that QD-Apt could penetrate the blood–brain barrier and then selectively accumulate in the tumors through binding to EGFRvIII, and consequently, generate a strong fluorescence, which contributed to the margins of gliomas that were visualized clearly, and thus, help the surgeons realize the maximum safe resection of glioma. In addition, QD-Apt can also be applied in preoperative diagnosis and postoperative examination of glioma. Therefore, these achievements facilitate the use of tumor-targeted fluorescence imaging in the diagnosis, surgical resection, and postoperative examination of glioma.

Dichlorofluorescein as a peroxidase mimic and its application to glucose detection

Herein, we discovered, for the first time, that 2′,7′-dichlorofluorescein (DCF), as a novel peroxidase mimic, can catalyze the oxidation of the classical peroxidase substrate 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H2O2, producing a colour reaction. Although a variety of peroxidase mimics have been actively developed recently, few studies have been focused on the small molecule peroxidase mimics. As a novel peroxidase mimic, DCF was verified to be highly stable, well reproducible and of low cost, exhibiting typical Michaelis–Menten kinetics and a strong affinity towards H2O2 and TMB. On the basis of these findings, we further demonstrated the novel use of DCF for easy and simple detection of H2O2 and glucose. This type of new small molecule peroxidase mimic is expected to be used in medical clinical diagnosis, and this novel finding not only further confirms the peroxidase activity of small molecules of fluorescein derivatives, but also opens new opportunities to deepen the knowledge of this new class of small molecule enzymes.

A sandwich immunoassay for detection of Aβ1-42 based on quantum dots

Alzheimers disease (AD) is the primary cause of dementia over the age of 60, affecting more than 35 million people worldwide. Methods for early diagnosis of AD are critical for the development of effective treatments to combat this debilitating disease. It was confirmed that amyloid-beta peptide 1-42 (Aβ(1-42)) is the biomarker of its early diagnosis. In this work, we present a new sandwich immunoassay method for the detection of Aβ(1-42) based on quantum dot (QDs) nanolabels and magnetic separation. In the presence of Aβ(1-42), QDs linked to magnetic beads (MB) via the formation of immune-sandwich complex and can be removed by a magnetic field. And as a result, fluorescence intensity from QDs in the supernatant decreased. Under the optimized conditions, there is a linear relationship between the fluorescence intensity of supernatant solution and the concentration of Aβ(1-42) from 0.50 to 8.0 nM with a limit detection of 0.2 nM (3σ). This immunoassay was applied to detect Aβ(1-42) in human cerebrospinal fluid (CSF) successfully.

Asn-Gly-Arg-modified polydopamine-coated nanoparticles for dual-targeting therapy of brain glioma in rats

The blood-brain barrier (BBB) is the major clinical obstacle in the chemotherapeutic management of brain glioma. Here we synthesized a pH-sensitive dual-targeting doxorubicin (DOX) carrier to compromise tumor endothelial cells, enhance BBB transportation, and improve drug accumulation in glioma cells. The drug delivery system was constructed with polydopamine (PDA)-coated mesoporous silica nanoparticles (NPs, MSNs) and the PDA coating was functionalized with Asn-Gly-Arg (NGR), a ligand with specific affinity for cluster of differentiation 13 (CD13). MSN-DOX-PDA-NGR showed a higher intracellular accumulation in primary brain capillary endothelial cells (BCECs) and C6 cells and greater BBB permeability than the non-targeting NPs (MSN-DOX-PDA) did in vitro. Ex vivo and in vivo tests showed that MSN-DOX-PDA-NGR had a higher accumulation in intracranial tumorous tissue than the undecorated NPs did. Furthermore, the antiangiogenesis and antitumor efficacy of MSN-DOX-PDA-NGR were stronger than that of MSN-DOX-PDA. Therefore, these results indicate that the dual-targeting vehicles are potentially useful in brain glioma therapy.

Intrinsic Peroxidase-like Activity of Ficin

Ficin is classified as a sulfhydryl protease isolated from the latex of fig trees. In most cases, a particular enzyme fits a few types of substrate and catalyzes one type of reaction. In this investigation, we found sufficient proofs for the intrinsic peroxidase-like activity of ficin and designed experiments to examine its effectiveness in a variety of scenarios. Ficin can transform peroxidase substrates to colored products in the existence of H2O2. Our results also indicate that the active sites of peroxidase-like activity of ficin are different from that of protease, which reveals that one enzyme may catalyze more than one kind of substrate to perform different types of reactions. On the basis of these findings, H2O2 releasing from MCF-7 cells was detected successfully. Our findings support a wider application of ficin in biochemistry and open up the possibility of utilizing ficin as enzymatic mimics in biotechnology and environmental monitoring.