Molecularly imprinted polymer-based SERS sensing of transferrin in human serum.

Specific detection of glycoproteins such as transferrin (TRF) related to neurological diseases, hepatoma and other diseases always plays an important role in the field of disease diagnosis. We designed an antibody-free immunoassay sensing method based on molecularly imprinted polymers (MIPs) formed by the polymerization of multiple functional monomers for the sensitive and selective detection of TRF in human serum. In the sandwich surface-enhanced Raman spectroscopy (SERS) sensor, the TRF-oriented magnetic MIP nanoparticles (Fe3O4@SiO2-MIPs) served as capture units to specifically recognize TRF and 4-mercaptophenylboronic acid-functionalized gold nanorods (MPBA-Au NRs) served as SERS probes to label the targets. In order to achieve stronger interaction between the recognition cavities of the prepared MIPs and the different amino acid fragments that make up TRF, Fe3O4@SiO2-MIPs were obtained through polycondensation reactions between more silylating reagents, enhancing the specific recognition of the entire TRF protein and achieving high IF. This sensing method exhibited a good linear response to TRF within the TRF concentration range of 0.01 ng mL-1 to 1 mg mL-1 (R2 = 0.9974), and the LOD was 0.00407 ng mL-1 (S/N = 3). The good stability, reproducibility and specificity of the resulting MIP based SERS sensor were demonstrated. The determination of TRF in human serum confirmed the feasibility of the method in practical applications.

Garlic-Derived Exosome-like Nanovesicles-Based Wound Dressing for Staphylococcus aureus Infection Visualization and Treatment.

Garlic-derived exosome-like nanovesicles (GELNs) could function in interspecies communication and may serve as natural therapeutics to regulate the inflammatory response or as nanocarriers to efficiently deliver specific drugs. Staphylococcus aureus (S. aureus) is able to hide within host cells to evade immune clearance and antibiotics, leading to life-threatening infections. On-site detection and efficient treatment of intracellular S. aureus infection in wounds remain challenging. Herein, we report a thermosensitive, injectable, visible GELNs-based wound dressing, Van@GELNs/F127 hydrogel (gel Van@GELNs), which is H2O2-responsive and can slowly release vancomycin into host cells forS. aureus infection visualization and treatment in wounds. GELNs show inherent antibacterial activity, which is significantly enhanced after loading vancomycin. Both GELNs and Van@GELNs have the ability to be internalized by cells, so Van@GELNs are more effective than free vancomycin in killing S. aureus in RAW 264.7 macrophages. When applied to an S. aureus-infected wound on a mouse, the colorless HRP&ABTS/Van@GELNs/F127 solution immediately changes to a green hydrogel and shows better therapeutic effect than vancomycin. Thus, direct visualization by the naked eye and effective treatment of S. aureus infection in wounds are achieved by gel Van@GELNs. We anticipate gel Van@GELNs be applied for the theranostics of S. aureus infection diseases in the clinic in the near future.

Electrochemiluminescence Analysis of Multiple Glycans on Single Living Cell with a Closed Bipolar Electrode Array Chip.

The profiling of multiple glycans on a single cell is important for elucidating glycosylation mechanisms and accurately identifying disease states. Herein, we developed a closed bipolar electrode (BPE) array chip for live single-cell trapping and in situ galactose and sialic acid detection with the electrochemiluminescence (ECL) method. Methylene blue-DNA (MB-DNA) as well as biotin-DNA (Bio-DNA) codecorated AuNPs were prepared as nanoprobes, which were selectively labeled on the cell surface through chemoselective labeling techniques. The individual cell was captured and labeled in the microtrap of the cathodic chamber, under an appropriate potential, MB molecules on the cellular membrane underwent oxidation, triggering the reduction of [Ru(bpy)3]2+/TPA and consequently generating ECL signals in the anodic chamber. The abundance of MB groups on the single cell enabled selective monitoring of both sialic acid and galactosyl groups with high sensitivity using ECL. The sialic acid and galactosyl content per HepG2 cell were detected to be 0.66 and 0.82 fmol, respectively. Through comprehensive evaluation of these two types of glycans on a single cell, tumor cells, and normal cells could be effectively discriminated and the accuracy of single-cell heterogeneous analysis was improved. Additionally, dynamic monitoring of variations in galactosyl groups on the surface of the single cell was also achieved. This work introduced a straightforward and convenient approach for heterogeneity analysis among single cells.

Enhanced Photoacoustic Imaging of Urokinase-Type Plasminogen Activator Activity in Tumors.

Photoacoustic (PA) imaging of urokinase-type plasminogen activator (uPA) activity in vivo holds high promise for early diagnosis of breast cancer. Molecular probes with resisted fluorescence (FL) emission for enhanced PA signals of uPA activity have not been reported. Herein, we proposed a molecular probe Cbz-Gly-Gly-Arg-Phe-Phe-IR775 (Z-GGRFF-IR775) which, upon uPA cleavage, assembled into nanoparticles FF-IR775-NP with quenched fluorescence but enhanced PA signals. Experimental results validated that, upon uPA activation, Z-GGRFF-IR775 exhibited 4.7-fold, 4.1-fold, and 2.9-fold higher PA signals over those in uPA inhibitor-treated control groups in vitro, in MDA-MB-231 cells, and in a tumor-bearing mouse model, respectively. We anticipate that this probe could be applied for highly sensitive PA imaging of uPA activity in early stage malignant tumors in the near future.

Synergistic ROS generation and directional overloading of endogenous calcium induce mitochondrial dysfunction in living cells.

Taking advantage of endogenous Ca2+ to upregulate intramitochondrial Ca2+ level has become a powerful mean for mitochondrial dysfunction-mediated tumor therapy. However, the Ca2+ entered into mitochondria is limited ascribing to the uncontrollability and non-selectivity of endogenous Ca2+ transport. It remains a great challenge to make the maximum use of endogenous Ca2+ to ensure sufficient Ca2+ overloading in mitochondria. Herein, we smartly fabricate an intracellular Ca2+ directional transport channel to selectively transport endogenous Ca2+ from endoplasmic reticulum (ER) to mitochondria based on cascade release nanoplatform ABT-199@liposomes/doxorubicin@FeIII-tannic acid (ABT@Lip/DOX@Fe-TA). In tumor acidic microenvironment, Fe3+ ions are firstly released and reduced by tannic acid (TA) to Fe2+ for ROS generation. Subsequently, under the NIR light irradiation, the released ABT-199 molecules combine with ROS contribute to the formation of IP3R-Grp75-VDAC1 channel between ER and mitochondria, thus Ca2+ ions are directionally delivered and intramitochondrial Ca2+ level is significantly upregulated. The synergetic ROS generation and mitochondrial Ca2+ overloading effectively intensifies mitochondrial dysfunction, thereby achieving efficient tumor inhibition. This work presents a new insight and promising avenue for endogenous Ca2+-involved tumor therapies.

Intramolecular fluorescence resonance energy transfer strategy for accurate detection of AFP-L3% and improved diagnosis of hepatocellular carcinoma.

Early and accurate diagnosis of hepatocellular carcinoma (HCC) is of significant importance for improving the survival rate and quality of life for HCC patients. The combined detection of alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3), namely AFP-L3%, can greatly improve the accuracy of HCC diagnosis compared with AFP detection. Herein, we developed a novel intramolecular fluorescence resonance energy transfer (FRET) strategy for sequential detection of AFP and AFP-specific core fucose to improve the diagnosis accuracy of HCC. Firstly, fluorescence-labeled AFP aptamer (AFP Apt-FAM) was used to specifically recognize all AFP isoforms, and total AFP was quantitatively determined using fluorescence intensity of FAM. Then, 4-((4-(dimethylamino)phenyl)azo)benzoic acid (Dabcyl) labeled lectins (PhoSL-Dabcyl) were used to specifically recognize the core fucose expressed on AFP-L3 that does not bind to other AFP isoforms. The combination of FAM and Dabcyl on the same AFP molecule could generate FRET effect, thereby quenching the fluorescence signal of FAM and quantitatively determining AFP-L3. After that, AFP-L3% was calculated according to the ratio of AFP-L3 to AFP. With this strategy, the concentration of total AFP, AFP-L3 isoform as well as the AFP-L3% were sensitively detected. Detection limits of 0.66 and 0.186 ng/mL were obtained for AFP and AFP-L3 in human serum, respectively. Clinical human serum test results showed that AFP- L3 % test was more accurate than AFP assay to distinguish healthy people, HCC patients and benign liver disease patients. Therefore, the proposed strategy is simple, sensitive and selective, which can improve the accuracy of early diagnosis of HCC, and has good clinical application potential.

Insight into Iron Leaching from an Ascorbate-Oxidase-like Fe-N-C Nanozyme and Oxygen Reduction Selectivity.

Ascorbate (H2 A) is a well-known antioxidant to protect cellular components from free radical damage and has also emerged as a pro-oxidant in cancer therapies. However, such "contradictory" mechanisms underlying H2 A oxidation are not well understood. Herein, we report Fe leaching during catalytic H2 A oxidation using an Fe-N-C nanozyme as a ferritin mimic and its influence on the selectivity of the oxygen reduction reaction (ORR). Owing to the heterogeneity, the Fe-Nx sites in Fe-N-C primarily catalyzed H2 A oxidation and 4 e- ORR via an iron-oxo intermediate. Nonetheless, trace O2 ⋅- produced by marginal N-C sites through 2 e- ORR accumulated and attacked Fe-Nx sites, leading to the linear leakage of unstable Fe ions up to 420 ppb when the H2 A concentration increased to 2 mM. As a result, a substantial fraction (ca. 40 %) of the N-C sites on Fe-N-C were activated, and a new 2+2 e- ORR path was finally enabled, along with Fenton-type H2 A oxidation. Consequently, after Fe ions diffused into the bulk solution, the ORR at the N-C sites stopped at H2 O2 production, which was the origin of the pro-oxidant effect of H2 A.

Molecular assembly of carbon nitride-based composite membranes for photocatalytic sterilization and wound healing.

Polymeric carbon nitride (pCN) has attracted increasing interest as a metal-free photocatalyst because of its high efficiency in reactive oxygen species (ROS) generation. However, due to poor solubility, compounding pCN at the molecular level into more advanced nanocomposites remains a challenge. Herein, we report the dissolution of pCN in polyphosphoric acid (PPA) for the first time and fluid-phase assembly with carbon nanotubes (CNTs) into a flexible free-standing membrane. Mechanism and generality studies disclosed that the coordination of the acidity, viscosity, and adsorption energy of the solvents led to the successful dissolution of pCN. Interestingly, the pCN/CNTs molecular composite membrane exhibited not only superior mechanical properties and cycling performance as a result of strengthened π-π interfacial interaction, but also outstanding inactivation of E. coli and S. aureus in sterilization and wound healing for laboratory mice via photogenerated oxygen radicals. It would open a new era of pCN for biomedical applications in molecular composite membranes, beyond the traditional solar fuel applications in powders.

Sensitive detection of PARP-1 activity by electrochemical impedance spectroscopy based on biomineralization.

Poly(ADP)ribose polymerase-1 (PARP-1) has attracted much attention as a tumor marker in recent years. Based on the large negative charge and hyperbranched structure of PARP-1 amplified products (PAR), many detection methods have been established. Herein, we proposed a label-free electrochemical impedance detection method based on the large amount of phosphate groups (PO43-) on the surface of PAR. Although EIS method has high sensitivity, it is not sensitive enough to discern PAR effectively. Therefore, biomineralization was incorporated to increase the resistance value (Rct) distinctly because of the poor electrical conductivity of CaP. During biomineralization process, plentiful Ca2+ was captured by PO43- of PAR through electrostatic interaction, resulting in an increasing Rct of modified ITO electrode. In contrast, when PRAP-1 was absent, only a little Ca2+ was adsorbed on the phosphate backbone of the activating dsDNA. As a result, the biomineralization effect was slight and only a negligible Rct change occurred. Experiment results showed that Rct was associated closely with the activity of PARP-1. There was a linear correlation between them when the activity value was in the range of 0.005-1.0 U. The calculated detection limit was 0.003 U. Results of real samples detection and the recovery experiments were satisfactory, indicating the method has an excellent application prospect.

Electrochemiluminescence nanoemitters for immunoassay of protein biomarkers.

The family of electrochemiluminescent luminophores has witnessed quick development since the electrochemiluminescence (ECL) phenomenon of silicon nanoparticles was first reported in 2002. Moreover, these developed ECL nanoemitters have extensively been applied in sensitive detection of protein biomarker by combining with immunological recognition. This review firstly summarized the origin and development of various ECL nanoemitters including inorganic and organic nanomaterials, with an emphasis on metal-organic frameworks (MOFs)-based ECL nanoemitters. Several effective strategies to amplify the ECL response of nanoemitters and improve the sensitivity of immunosensing were discussed. The application of ECL nanoemitters in immunoassay of protein biomarkers for diagnosis of cancers and other diseases, especially lung cancer and heart diseases, was comprehensively presented. The recent development of ECL imaging with the nanoemitters as ECL tags for detection of multiplex protein biomarkers on single cell membrane also attracted attention. Finally, the future opportunities and challenges in the ECL biosensing field were highlighted.

Efficiently targeted therapy of glioblastoma xenograft via multifunctional biomimetic nanodrugs.

BACKGROUND: Glioblastoma multiforme (GBM) is a fatal malignant primary brain tumor in adults. The therapeutic efficacy of chemotherapeutic drugs is limited due to the blood-brain barrier (BBB), poor drug targeting, and short biological half-lives. Multifunctional biomimetic nanodrugs have great potential to overcome these limitations of chemotherapeutic drugs. METHODS: We synthesized and characterized a biomimetic nanodrug CMS/PEG-DOX-M. The CMS/PEG-DOX-M effectively and rapidly released DOX in U87 MG cells. Cell proliferation and apoptosis assays were examined by the MTT and TUNEL assays. The penetration of nanodrugs through the BBB and anti-tumor efficacy were investigated in the orthotopic glioblastoma xenograft models. RESULTS: We showed that CMS/PEG-DOX-M inhibited cell proliferation of U87 MG cells and effectively induced cell apoptosis of U87 MG cells. Intracranial antitumor experiments showed that free DOX hardly penetrated the BBB, but CMS/PEG-DOX-M effectively reached the orthotopic intracranial tumor through the BBB and significantly inhibited tumor growth. Immunofluorescence staining of orthotopic tumor tissue sections confirmed that nanodrugs promoted apoptosis of tumor cells. This study developed a multimodal nanodrug treatment system with the enhanced abilities of tumor-targeting, BBB penetration, and cancer-specific accumulation of chemotherapeutic drugs by combining chemotherapy and photothermal therapy. It can be used as a flexible and effective GBM treatment system and it may also be used for the treatment of other central nervous systems (CNS) tumors and extracranial tumors.

Sensitive detection of organophosphorus pesticides based on the localized surface plasmon resonance and fluorescence dual-signal readout.

In this work, a dual-signal visual biosensor was designed for organophosphorus pesticides (OPs) detection using DNA functionalized Ag/Au bimetallic nanoparticles (Ag/Au NPs) as multifunctional nanoprobe. The dual-signal detection strategy was based on the inhibition of enzyme-induced H2O2 generation by OPs in the detection solution containing acetylcholinesterase (AChE), choline oxidase (CHO), acetylcholine (ACh) and nanoprobe. H2O2 produced by enzyme-catalyzed reaction could trigger the etching of Ag and dissociation of carboxyfluorescein (FAM)-labeled aptamer from the nanoprobe, resulting in significant localized surface plasmon resonance (LPRR) and fluorescence (FL) signal responses. In the presence of OPs, AChE activity was inhibited to disrupt the enzymatic generation of H2O2, which allowed to simultaneous quantitative measure OPs through the LSPR peak shifts and FL intensity variations of the nanoprobe. The LSPR/FL dual-signal biosensor showed great selectivity and sensitivity for OPs detection. In addition, two distinct colour changes were visually observed to match the LSPR/FL spectra signal responses, which was a feasible means for visual analysis of OPs. Consequently, the work provided a dual-signal visual biosensor via the combination of multifunctional nanoprobe, and had significant potential to monitor pesticide residue with high anti-interference capability and detection accuracy.

2D Copper(II) Metalated Metal-Organic Framework Nanocomplexes for Dual-enhanced Photodynamic Therapy and Amplified Antitumor Immunity.

The immunosuppressive tumor microenvironment (TME) poses tremendous challenges for efficient immunotherapy. Smart nanomedicine is designed to modulate immunosuppressive TMEs based on the combination of dual-enhanced photodynamic therapy (PDT) triggered immunogenic cell death (ICD) and relieved hypoxic microenvironment. Copper(II) metalated metal-organic framework nanosheets (Cu-TCPP(Al)) are the foundation of the nanomedicine, and platinum nanoparticles (Pt NPs) and folate are subsequently introduced onto the Cu-TCPP(Al) surface (Cu-TCPP(Al)-Pt-FA). Upon targeted cellular uptake, intracellular GSH concentration is decreased because of the specific adsorption between GSH and CuII; meanwhile, Pt NPs possess catalase-like activity, which can continuously depose intracellular H2O2 to O2 to alleviate the hypoxic TME. The two factors synergistically improve the ROS concentration for dual-enhanced PDT. The highly toxic ROS can correspondingly cause amplified oxidative stress and then trigger the ICD. The ICD process stimulates antigen-presenting cells and activates the systemic antitumor immune response. Furthermore, the relieved hypoxic TME increases the infiltration of cytotoxic T lymphocytes (CTLs) at the tumor site, which can promote the transformation of the immunosuppressive M2 macrophage to immunoactive M1 phenotype. The easily prepared yet versatile nanomedicine possesses an excellent antitumor effect with the cooperation of dual-enhanced PDT and immunotherapy.

Extended Conjugation Tuning Carbon Nitride for Non-sacrificial H2 O2 Photosynthesis and Hypoxic Tumor Therapy.

Artificial photocatalysis offers a clean approach for producing H2 O2 . However, the poor selectivity and activity of H2 O2 production hamper traditional industrial applications and emerging photodynamic therapy (PDT)/chemodynamic therapy (CDT). Herein, we report a C5 N2 photocatalyst with a conjugated C=N linkage for selective and efficient non-sacrificial H2 O2 production in both normoxic and hypoxic systems. The strengthened delocalization of π-electrons by linkers in C5 N2 downshifted the band position, thermodynamically eliminating side H2 evolution reaction and kinetically promoting water oxidation. As a result, C5 N2 had a competitive solar-to-chemical conversion efficiency of 0.55 % in overall H2 O2 production and exhibited by far the highest activity under hypoxic conditions (698 µM h-1 ). C5 N2 was further applied to hypoxic PDT/CDT with outstanding performance in apparent cancer cell death and synchronous bioimaging. The study sheds light on the photosynthesis of H2 O2 by carbon nitrides for health applications.

TfR Aptamer Enhanced Blood-Brain Barrier Penetration of Biomimetic Nanocomplexes for Intracellular Transglutaminase 2 Imaging and Silencing in Glioma.

Engineering a versatile nanocomplex integrating effective penetration of the blood-brain barrier (BBB), accurate diagnosis, and boosting therapy has always been an intractable challenge in glioblastoma multiforme (GBM). Herein, biomimetic nanocomplexes (TMPsM) for single intracellular transglutaminase 2 (TG2)-triggered self-assembly imaging and RNAi therapy for GBM are subtly developed. To prove the concept, transferrin receptor (TfR) aptamer-modified brain metastatic tumor cell membrane is prepared as the shell for dual BBB targeting capability and prolonged blood retention time. Upon targeting entering into GBM, hollow MnO2 is decomposed to release KKGKGQQ-tetraphenylethene (Pep-TPE) and siRNA. Owing to TG2 dependence, the non-emissive Pep-TPE would be self-aggregated to induce the emission turn-on in GBM that contain overexpressed TG2. The resulting aggregation-induced emission fluorescence imaging with a high signal-to-noise ratio can achieve the precise localization of the tumor and dynamic detection of TG2 activity, thereby allowing the GBM accurate diagnosis. Notably, the TG2 can be silenced by the released siRNA to cause cell apoptosis and increase chemotherapeutic sensitivity, ultimately realizing excellent antitumor efficacy. In vitro and in vivo results demonstrate that the as-prepared TMPsM indeed possess superior BBB penetration, precise diagnosis, and effective therapy of GBM. The proposed strategy may pioneer a new path for the theranostics of brain tumors.

Fluorescence imaging of FEN1 activity in living cells based on controlled-release of fluorescence probe from mesoporous silica nanoparticles.

Flap endonuclease 1 (FEN1) is a structure-specific nuclease, which catalyzes the removal of 5' overhanging DNA flap from a specific DNA structure. FEN1 has been considered as an important biomarker for cancer diagnosis since it is over-expressed in various types of human tumor cells and closely related to cancer development. Nanoprobes gradually become basic tools for analyzing biomarkers variations in vivo. Here, we utilized aminoated mesoporous silica nanoparticles (NH2-MSNs) with a rich porous structure as the fluorescence nanoprobes to entrap the rhodamine 6G (Rh6G) molecules. Then gold nanoparticles linked specific single-stranded DNA (AuNPs-ssDNA) as a molecular gate was used to coat the NH2-MSNs surface. The fluorescence signal was weak when the fluorescence molecules were blocked by the AuNPs-ssDNA. In the presence of FEN1, it recognized and cleaved the specific ssDNA to release the Rh6G from NH2-MSNs, which resulted in recovered fluorescence signals. Thus, the sensitive detection of FEN1 activity was realized by controlled-release of Rh6G. The fluorescence signal showed a good linear relationship with the logarithm of FEN1 activity ranging from 0.05 to 1.75 U with a detection limit of 0.03 U. Moreover, confocal imaging demonstrated that the proposed biosensor could distinguish tumor cells from normal cells. Therefore, this technique contributes to clinical diagnostic and therapeutic monitoring.

In Situ Imaging of Endogenous Hydrogen Peroxide Efflux from Living Cells via Bipolar Gold Nanoelectrode Array and Electrochemiluminescence Technology.

The integration of a closed bipolar electrode (c-BPE) array and electrochemiluminescence (ECL) detection received a boost in applications in the detection of cell adhesion and disease-related biomarkers. This work proposed a gold nanorod array based c-BPE-ECL system to realize an in situ image of endogenous hydrogen peroxide (H2O2) efflux from living cells and parallel analysis of endogenous H2O2 released from multiple cells by converting electrochemical signals into optical signals. The gold nanorod array with high density was prepared by a repeating chronopotentiometry procedure with anodic aluminum oxide (AAO) membrane as a template. The c-BPE array was fabricated by assembling poly(dimethylsiloxane) (PDMS) chips on both sides of the gold nanorod array. When an appropriate driving potential is applied, H2O2 generated from living cells at the sensing pole was reduced on the gold nanorod, triggering the oxidation of the ECL reagent at the reporting pole, which allowed the detection of H2O2 released from living cells. Under phorbol myristate acetate (PMA) stimulation, H2O2 released from living HeLa, HepG2, MCF-7, and LO2 cells was determined to be 47, 32.4, 25.7, and 6.3 µM, respectively. This indicated that the amount of H2O2 released from PMA-stimulated cancer cells was significantly higher than that from the stimulated normal cells. This work presented a new approach for in situ imaging of H2O2 released from living cells and could also be used to detect other electrochemically active or non-electrochemically active molecules through simple cell surface modification, which may have potential applications in cell apoptosis study and disease diagnosis.

Three-Dimensional Microfluidic Chip for Efficient Capture of Secretory Autophagosomes and Sensitive Detection of Their Surface Proteins.

Recent studies on autophagy demonstrated a new extracellular secretion pathway for autophagosomes in addition to the routinely described intracellular degradation pathway. Besides, the secretory autophagosomes were found closely related to the occurrence and development of cancers. Therefore, analysis of the protein expression on secretory autophagosomes is a promising noninvasive strategy for cancer diagnosis and mechanism study. Herein, we constructed a three-dimensional (3D) microfluidic chip employing a fusiform micropillar array and layer-by-layer modification of gelatins, which obviously enhanced the mass transfer between reactants and increased the immobilization sites for capture antibody. As a result, the autophagosome capture efficiency of the 3D chip (74%) is significantly higher than that of the unmodified flat chip (47%). Using a two-step immunoreaction, ovarian cancer cell-secreted autophagosomes were successfully captured and detected. The results showed that two proteins, LC3B and HSP60 at the surface of autophagosomes, can be detected with limits of detection (LODs) of 141 particles µL-1 and 126 particles µL-1, respectively. In addition, both LC3B and HSP60 expressions on autophagosomes can be used to distinguish the serum samples between cancer patients and healthy people, with a p value less than 0.01 (statistically significant difference) or 0.05 (statistically different), respectively. Moreover, the summed signal of LC3B and HSP60 showed a p value less than 0.001 (extremely statistically significant difference), demonstrating the good potential of this chip for further application in cancer diagnosis.

Gold Nanowires Array-Based Closed Bipolar Nanoelectrode System for Electrochemiluminescence Detection of α-Fetoprotein on Cell Surface.

Inspired by the promising applications of a closed bipolar electrodes (c-BPEs) system in electrochemiluminescence (ECL) detection of cell adhesion and disease-related biomarkers, here, a gold nanowires array-based c-BPEs system was constructed for cell surface protein detection. Regular and uniform gold nanowires array were prepared by intermittent potentiostatic deposition. Then, two poly(dimethylsiloxane) (PDMS) chips with a hole diameter of 2 mm as a reservoir were placed at both sides of Au nanowires array to construct c-BPEs system. Thionine-functionalized silicon dioxide nanoparticles conjugated to antibody (Ab2-Th@SiO2) were used as the electrochemical probe, while [Ru(bpy)3]2+-wrapped SiO2 nanoparticles (Ru(II)@SiO2) were employed as the ECL signal readout. Taking α-fetoprotein (AFP) as model, the gold nanowires array-based c-BPEs system allowed sensitive detection of AFP at a linear range from 0.002 to 50.0 ng/mL and at least 6 living cells ascribing to the synergetic amplification effect at both sensing and reporting chambers. Besides, the amount of AFP expressed by HepG2 cells was calculated to be 6.71 pg/cell. The presented strategy with high sensitivity provided a promising and universal platform for the detection of other cancer cells and disease-related biomarkers (such as proteins, glycan, miRNA).

Cascaded Nanozyme System with High Reaction Selectivity by Substrate Screening and Channeling in a Microfluidic Device.

Surpassing natural enzymes in cost, stability and mass production, nanozymes have attracted wide attention in fields from disease diagnosis to tumor therapy. However, nanozymes intrinsically have low reaction selectivity, which significantly restricts their applications. A general method is reported to address this challenge by following a biomimetic operation principle of substrates channeling and screening. Two oxidase- and peroxidase-like nanozymes (i.e., emerging N-doped carbon nanocages and Prussian blue nanoparticles), were cascaded as a proof of concept to improve the reaction selectivity in transforming the substrate into the targeted product by more than 2000 times. The cascaded nanozymes were also adopted to a spatially confined microfluidic device, leading to more than 100-fold enhancement of the reaction efficiency due to signal amplification.