Engineering Assembly of Plasmonic Virus-Like Gold SERS Nanoprobe Guided by Intelligent Dual-Machine Nanodevice for High-Performance Analysis of Tetracycline.

Accurate detection of trace tetracyclines (TCs) in complex matrices is of great significance for food and environmental safety monitoring. However, traditional recognition and amplification tools exhibit poor specificity and sensitivity. Herein, a novel dual-machine linkage nanodevice (DMLD) is proposed for the first time to achieve high-performance analysis of TC, with a padlock aptamer component as the initiation command center, nucleic acid-encoded multispike virus-like Au nanoparticles (nMVANs) as the signal indicator, and cascade walkers circuit as the processor. The existence of spike vertices and interspike nanogaps in MVANs enables intense electromagnetic near-field focusing, allowing distinct surface-enhanced Raman scattering (SERS) activity. Moreover, through the sequential activation between multistage walker catalytic circuits, the DLMD system converts the limited TC recognition into massive engineering assemblies of SERS probes guided by DNA amplicons, resulting in synergistic enhancement of bulk plasmonic hotspot entities. The continuously guaranteed target recognition and progressively promoted signal enhancement ensure highly specific amplification analysis of TC, with a detection limit as low as 7.94 × 10-16 g mL-1. Furthermore, the reliable recoveries in real samples confirm the practicability of the proposed sensing platform, highlighting the enormous potential of intelligent nanomachines for analyzing the trace hazards in the environment and food.

An "off-on-off" type electrochemical biosensor for detecting multiple biomarkers with DNAzyme-mediated entropy-driven catalytic and DSN enzyme-assisted recycling amplification.

In this work, an intelligent and versatile electrochemical biosensor was constructed to detect two types of biomarkers by utilizing "off-on-off" switching. Firstly, human apurinic/apyrimidinic endonuclease1(APE1) mediated specific cleavage of the AP site, initiating activation DNAzyme and entropy-driven catalytic (EDC) reaction. Subsequently, large amounts of ferrocene labeled single-stranded DNA was released and captured with a remarkable electrochemical signal, achieving "off-on" state. In the presence of microRNA 21(miRNA-21), the DNA/RNA heteroduplexes were formed and cleaved by duplex-specific nuclease (DSN) with recovery the target miRNA-21, causing the current suppression in an "on-off" state. This sensor achieved highly sensitive detection of APE1 and miRNA-21 with a detection limit of 2.5 mU·mL-1 and 1.33 × 10-20 M, respectively, and also exhibited good selectivity, reproducibility and stability. Moreover, this proposed biosensor made it possible to realize analysis of multiple types of biomarkers on a single sensor, which improved utilization and analysis efficiency compared to traditional sensors. This study might open a new avenue to design multifunctional sensing platform for biological research and early disease diagnosis.

Polyhedral Au Nanoparticle/MoOx Heterojunction-Enhanced Ultrasensitive Dual-Mode Biosensor for miRNA Detection Combined with a Nonenzymatic Cascade DNA Amplification Circuit.

A novel homologous surface-enhanced Raman scattering (SERS)-electrochemical (EC) dual-mode biosensor based on a 3D/2D polyhedral Au nanoparticle/MoOx nanosheet heterojunction (PAMS HJ) and target-triggered nonenzyme cascade autocatalytic DNA amplification (CADA) circuit was constructed for highly sensitive detection of microRNA (miRNA). Mixed-dimensional heterostructures were prepared by in situ growth of polyhedral Au nanoparticles (PANPs) on the surface of MoOx nanosheets (MoOx NSs) via a seed-mediated growth method. As a detection substrate, the resulting PAMS HJ shows the synergistic effects of both electromagnetic and chemical enhancements, efficient charge transfer, and robust stability, thus achieving a high SERS enhancement factor (EF) of 4.2 × 109 and strong EC sensing performance. Furthermore, the highly efficient molecular recognition between the target and smart lock probe and the gradually accelerated cascade amplification reaction further improved the selectivity and sensitivity of our sensing platform. The detection limits of miRNA-21 in SERS mode and EC mode were 0.22 and 2.69 aM, respectively. More importantly, the proposed dual-mode detection platform displayed excellent anti-interference and accuracy in the analysis of miRNA-21 in human serum and cell lysates, indicating its potential as a reliable tool in the field of biosensing and clinical analysis.

CuS quantum dots activated DNAzyme for ratiometric electrochemical detection of telomerase activity.

Telomerase activity detection has attracted much attention concerning its importance for early cancer diagnosis. Here, we established a ratiometric electrochemical biosensor for telomerase detection based on CuS quantum dots (CuS QDs) dependent DNAzyme-regulated dual signals. The telomerase substrate probe was used as the linker to combine the DNA fabricated magnetic beads and CuS QDs. In this way, telomerase extended the substrate probe with repeated sequence to from hairpin structure, releasing CuS QDs as an input to DNAzyme modified electrode. DNAzyme was cleaved with high current of ferrocene (Fc) and low current of methylene blue (MB). On the basis of the obtained ratiometric signals, telomerase activity detection was achieved in the range of 1.0 × 10-12-1.0 × 10-6 IU/L, with the limit of detection down to 2.75 × 10-14 IU/L. Moreover, telomerase activity from HeLa extracts was also tested to verify the clinical application.

Pollution heaven or pollution halo? Assessing the role of heterogeneous environmental regulation in the impact of foreign direct investment on green economic efficiency.

As the main engine of the global economy, China has been attracting increasing foreign direct investment (FDI) since the 1980s. The frequent occurrence of pollution incidents by multinational companies and the continuous deterioration of the environment have prompted China to attach importance to environmental regulations and attempt to avoid the potential pollution heaven effect of FDI on green development. To assess the effectiveness of these environmental regulations, this paper investigates the moderating effect of environmental regulation, in particular, the heterogeneous environmental regulatory tools, on the relationship between FDI and green economic efficiency. In addition, the spatial performance of these moderating effects is examined through the spatial Durbin model (SDM), using China's provincial panel data from 2004 to 2018. The results show that environmental regulation has an overall positive moderating effect, exacerbating the pollution heaven effect of FDI on green economic efficiency. In the meantime, the moderating effects of heterogeneous environmental regulations are obviously different; i.e., command-and-control and public-participation-based environmental regulations have positive moderating effects, while market-based environmental regulation has a negative moderating effect. In addition, in terms of spatial performance, the market-based environmental regulation has a positive spillover effect, thereby promoting green economic efficiency in surrounding regions, which is contrary to command-and-control and public-participation-based environmental regulations. Based on the above findings, this paper makes some recommendations for policymakers.

DNA-Au Janus Nanoparticles for In Situ SERS Detection and Targeted Chemo-photodynamic Synergistic Therapy.

Cancer theranostics is of great significance in the personalized therapy. In this work, stable Janus nanoparticles (JNPs) containing PEG and two kinds of DNAs were prepared by means of "click chemistry". In response to ATP or acid condition, the prepared JNPs could form Au NP dimers, which facilitate in situ SERS detection and SERS imaging analysis of cancer cells due to the formation of "hot spots" in the nanogap between the Au NP dimers. A detection limit of 2.3 × 10-9 M was obtained for ATP. As for a pH sensor, the SERS signals increased with the decrease of pH value from 8.0 to 4.0. In situ monitoring of ATP or acid condition in cancer cells by SERS can improve the accuracy and sensitivity of diagnosis. Moreover, drugs and photosensitizers loaded on the other side of JNPs led to the chemotherapy/photodynamic therapy synergistic antitumor effect, which was verified by in vitro and in vivo experiments. Given the excellent performance in SERS detection and cancer therapy, the developed JNPs hold considerable potential in cancer theranostics.

One-pot alkali cutting-assisted synthesis of fluorescence tunable amino-functionalized graphene quantum dots as a multifunctional nanosensor for sensing of pH and tannic acid.

Herein, a one-pot alkali cutting-assisted synthesis approach has been developed to gain fluorescence (FL) tunable amino functionalized GQDs (NH2-GQDs), which exhibit concentration- and excitation-dependent FL behaviors, due to the self-assembled J-type aggregation effect and different electronic transitions governed by graphene basal plane and functional groups. While NH2-GQDs possess brighter FL emission than pristine GQDs, owning to the functionalization of amino groups with strong electron withdrawing ability. Particularly, the pH-dependent FL behavior of NH2-GQDs further reflects the FL emission mechanism originated from the intrinsic zigzag sites and introduced amino and carboxylic groups, which is available for pH sensing. Moreover, the NH2-GQDs also show a FL quenching upon reaction with tannic acid (TA), resulting in the construction of a FL turn-off TA sensing platform. A good linear relationship is obtained between logarithm of FL intensity (log F) and TA concentration in a linear dynamic range of 1-40 µM and a limit of detection of 43.3 nM (3σ/s, n = 9) is achieved, with a precision of 0.08% RSD at a concentration level of 5 µM (n = 9). This work features a simple and direct approach to acquire multifunctional nanosensor, providing great potential for further applications in chem/biosensing.

Self-Assembly of Multifunctional DNA Nanohydrogels with Tumor Microenvironment-Responsive Cascade Reactions for Cooperative Cancer Therapy.

A DNA structure-based nanoreactor has emerged as a promising biomaterial for antitumor therapy with its intrinsic biodegradability, biocompatibility, and tunable multifunctionality. Herein, the intelligent DNA nanohydrogel was reported to target cancer cells, control the size, be pH-responsive, and be loaded with glucose oxidase (GOx). Two kinds of X-shaped DNA monomers and DNA linkers were assembled to form a DNA nanohydrogel by hybridization. GOx was successfully encapsulated in the DNA nanohydrogel. The DNA linker was designed with i-motif sequences and modified with ferrocene (Fc). The i-motif-like quadruplex structures were formed in acidic tumor microenvironments, resulting in the disassembly of the DNA nanohydrogel to release GOx. The GOx could oxidize the intratumoral glucose to produce gluconic acid and H2O2. The generated H2O2 was catalyzed by Fc to induce toxic hydroxyl radicals (•OH), which could effectively kill cancer cells. Both the in vitro and the in vivo results demonstrated that the multifunctional DNA nanohydrogel had high-efficiency tumor suppression through combined chemodynamic and starvation cancer therapies.

Dual-mode glucose nanosensor as an activatable theranostic platform for cancer cell recognition and cascades-enhanced synergetic therapy.

Integration of disease diagnosis and therapy is crucial in precise medicine, while the "always on" mode often hinders its clinical applications. Herein, inspired by cascaded catalysis, an integrated dual-mode glucose nanosensor as an activable theranostic platform is developed, which is further exploited for cancer cell recognition and enhanced synergistic therapy of lymph cancer. This nanosensor is prepared through the in-situ growth of silver nanoparticles (AgNPs) with the synergetic reduction of tannic acid (TA) and graphene quantum dots (GQDs), which are further decorated with glucose oxidase (GOx). A cascaded catalytic reaction is triggered by glucose, in which GOx catalyzes the oxidation of glucose into gluconic acid and hydrogen peroxide (H2O2), and hydroxyl radical (•OH) is further produced with the catalysis of GQDs nanozyme with peroxidase-like activity, resulting in the degradation of AgNPs@GQDs-GOx with the release of Ag+. Accordingly, a "turn-off" colorimetric and "turn-on" fluorescence dual-mode glucose nanosensor is fabricated, which is readily applied for cancer cell recognition via fluorescence imaging based on the high glucose level in tumor microenvironment. Moreover, the degradation of AgNPs@GQDs-GOx in response to glucose facilitates the cascades-enhanced synergistic therapy of lymph cancer with the combination of starving-like therapy, metal ion therapy and TA-induce apoptosis. This study highlights a glucose-activated theranostic nanoplatform, which provides a great opportunity for cancer-related biosensing, bioimaging and biomedical applications.

Many Birds, One Stone: A Smart Nanodevice for Ratiometric Dual-Spectrum Assay of Intracellular MicroRNA and Multimodal Synergetic Cancer Therapy.

The development of a theragnostic platform integrating precise diagnosis and effective treatment is significant but still extremely challenging. Herein, an integrated smart nanodevice composed of Au@Cu2-xS@polydopamine nanoparticles (ACSPs) and fuel DNA-conjugated tetrahedral DNA nanostructures (fTDNs) was constructed, in which the ACSP nanoprobe played multiple key roles in antitumor therapy as well as in situ monitoring of microRNAs (miRNAs) in cancer cells. Regarding the analysis, the ACSP probe contained two optical properties: excellent surface-enhanced Raman scattering (SERS) enhancement and high fluorescence (FL) quenching performance. Employing the ACSPs as the high-efficiency detection substrate combined with the fTDN-assisted DNA walking nanomachines as the superior amplification strategy, a SERS-FL dual-spectrum biosensor was constructed, which achieved an ultralow background signal and excellent sensitivity with detection limits of 0.11 pM and 4.95 aM by FL and SERS, respectively. Moreover, the rapid FL imaging and precise SERS quantitative detection for miRNA in cancer cells were also achieved by dual-signal ratio strategy, improving the accuracy of diagnosis. Regarding the therapeutic application, due to the high reactive oxygen species generation ability and excellent photothermal conversion efficiency, the ACSPs can also act as an all-in-one nanoagent for multimodal collaborative tumor therapy. Significantly, both in vivo and in vitro experiments confirmed its high biological safety and strong anticancer effect, indicating its promising theragnostic applications.

Rolling Circle Replication for Biosensing, Bioimaging, and Biomedicine.

Rolling circle replication (RCR), including rolling circle amplification (RCA) and rolling circle transcription (RCT), is an isothermal enzymatic reaction. Because of its high amplification efficiency, RCR is a powerful biosensing tool for detecting biomolecules. In recent years, RCR has also been extended to the field of bioimaging to better understand biological pathways. Furthermore, RCR provides a simple technique to design and generate DNA/RNA structures with unique advantages in delivering drugs and enhanced targeting ability. In this review, we introduce the fundamentals of RCR and describe the most recent advances in RCR-based detection methods and delivery vehicles for biosensing, bioimaging, and biomedicine. Finally, some challenges and further opportunities of RCR-based biotechnology are discussed.

Versatile electrochemical biosensor based on bi-enzyme cascade biocatalysis spatially regulated by DNA architecture.

The regulation of biocatalytic cascades in microenvironments for high performance and extended applications is still challenging. Herein, we develop a rolling circle amplification (RCA)-based one-pot method to prepare the micron-sized DNA flowers (DFs), which achieve the co-encapsulation and spatial regulation of bi-enzyme molecules, glucose oxidase (GOx) and horseradish peroxidase (HRP). In this system, GOx and HRP are integrated into the DFs simultaneously during RCA with the bridging of magnesium between enzyme residues and phosphate backbones on DFs. The cascade of GOx/HRP is regulated with the formation of highly ordered and hydrogen-bonded water environment in the cavity of DFs, resulting in an enhanced cascade catalytic efficiency compared with that in homogeneous solution. Moreover, the high density of DNA scaffold ensures the encapsulation of GOx/HRP with high efficiency. Accordingly, a glucose electrochemical biosensor with amplified signal response is fabricated using the as-prepared GOx/HRP DFs as biosensing interface, realizing sensitive detection of glucose. Further, through designing the complementary sequence of aptamer into the programmable circular template of RCA, the bi-enzyme co-encapsulated DFs are versatilely applied to sensitive and selective detection of cancerous exosomes and thrombin in "signal-on" and "signal-off" modes, respectively, which are further applied to the analysis of complex biological samples successfully. Overall, the encapsulation of multi-enzyme with DFs proposes a promising strategy to regulate the microenvironment of biocatalytic cascades, which hold great potential in biotechnology, bioanalysis and disease diagnosis.

Recent advances in templated synthesis of metal nanoclusters and their applications in biosensing, bioimaging and theranostics.

As a kind of promising nanomaterials, metal nanoclusters (MNCs) generally composed of several to hundreds of metal atoms have received increasing interest owing to their unique properties, such as ultrasmall size (<2 nm), fascinating physical and chemical properties, and so on. Recently, template-assisted synthesis of MNCs (e.g., Au, Ag, Cu, Pt and Cd) has attracted extensive attention in biological fields. Up to now, various templates (e.g., dendrimers, polymers, DNAs, proteins and peptides) with different configurations and spaces have been applied to prepare MNCs with the advantages of facile preparation, controllable size, good water-solubility and biocompatibility. Herein, we focus on the recent advances in the template-assisted synthesis of MNCs, including the templates used to synthesize MNCs, and their applications in biosensing, bioimaging, and disease theranostics. Finally, the challenges and future perspectives of template-assisted synthesized MNCs are highlighted. We believe that this review could not only arouse more interest in MNCs but also promote their further development and applications by presenting the recent advances in this area to researchers from various fields, such as chemistry, material science, physiology, biomedicine, and so on.

Cancer cell-targeted nanoprobe for multilayer imaging of diverse biomarkers and precise photodynamic therapy.

A novel multifunctional nanoprobe was designed for cancer cell targeted multilayer imaging of two cancer biomarkers. Based on the proposed method, in situ imaging of membrane MUC1 mucin and cytoplasmic microRNA miR-21 coupled with precise photodynamic therapy was achieved.

pH-responsive DNA nanomicelles for chemo-gene synergetic therapy of anaplastic large cell lymphoma.

Chemo-gene therapy is an emerging synergetic modality for the treatment of cancers. Herein, we developed pH-responsive multifunctional DNA nanomicelles (DNMs) as delivery vehicles for controllable release of doxorubicin (Dox) and anaplastic lymphoma kinase (ALK)-specific siRNA for the chemo-gene synergetic therapy of anaplastic large cell lymphoma (ALCL). Methods: DNMs were synthesized by performing in situ rolling circle amplification (RCA) on the amphiphilic primer-polylactide (PLA) micelles, followed by functionalization of pH-responsive triplex DNA via complementary base pairing. The anticancer drug Dox and ALK-specific siRNA were co-loaded to construct Dox/siRNA/DNMs for chemo-gene synergetic cancer therapy. When exposed to the acidic microenvironment (pH below 5.0), C-G·C+ triplex structures were formed, leading to the release of Dox and siRNA for gene silencing to enhance the chemosensitivity in ALCL K299 cells. The chemo-gene synergetic anticancer effect of Dox/siRNA/DNMs on ALCL was evaluated in vitro and in vivo. Results: The pH-responsive DNMs exhibited good monodispersity at different pH values, good biocompatibility, high drug loading capacity, and excellent stability even in the human serum. With the simultaneous release of anticancer drug Dox and ALK-specific siRNA in response to pH in the tumor microenvironment, the Dox/siRNA/DNMs demonstrated significantly higher treatment efficacy for ALCL compared with chemotherapy alone, because the silencing of ALK gene expression mediated by siRNA increased the chemosensitivity of ALCL cells. From the pathological analysis of tumor tissue, the Dox/siRNA/DNMs exhibited the superiority in inhibiting tumor growth, low toxic side effects and good biosafety. Conclusion: DNMs co-loaded with Dox and ALK-specific siRNA exhibited significantly enhanced apoptosis of ALCL K299 cells in vitro and effectively inhibited tumor growth in vivo without obvious toxicity, providing a potential strategy in the development of nanomedicines for synergetic cancer therapy.

Detection of DNA methyltransferase activity using template-free DNA polymerization amplification based on aggregation-induced emission.

A sensitive and selective fluorescence assay for DNA methyltransferase (MTase) activity detection was designed based on aggregation-induced emission (AIE) and target initiated template-free DNA polymerization. Quaternized tetraphenylethene salt was synthesized as the AIE probe, which binds to single-stranded DNA by electrostatic interaction. A hairpin probe was designed with a specific sequence for DNA MTase. In the presence of DNA MTase, the methylation reaction initiated DNA polymerization with terminal deoxynucleotidyl transferase (TdT), which activated the fluorescence intensity through AIE. The designed DNA sensor displayed a linear response to concentrations of DNA adenine methyltransferase (Dam) MTase from 0.5 U·mL-1 to 100 U mL-1, with a limit of detection of 0.16 U mL-1. The assay was also effective for detection of DNA MTase activity in human serum and for showing the inhibitory effect of 5-fluorouracil on Dam MTase.

An CuInS2 photocathode for the sensitive photoelectrochemical determination of microRNA-21 based on DNA-protein interaction and exonuclease III assisted target recycling amplification.

A photocathode is described for the determination of microRNA-21 by using CuInS2 as an active photocathode material. Exonuclease III assisted target recycling amplification was employed to enhance the detection sensitivity. The TATA-binding protein (TBP) was applied to enhance steric hindrance which decreases the photoelectrochemical intensity. This strategy is designed by combining the anti-interference photocathode material, enzyme assisted target recycling amplification and TBP induced signal off, showing remarkable amplification efficiency. Under the optimized conditions, the detection limit for microRNA-21 is as low as 0.47 fM, and a linear range was got from 1.0 × 10-15 M to 1.0 × 10-6 M. Graphical abstract Schematic representation of sensitive photoelectrochemical detection of microRNA-21.CuInS2 is used as an active photocathode material. Combined Exonuclease III assisted target recycling amplification and TATA-binding protein decreased of photoelectrochemical intensity, the detection limit was 0.47 fM with good selectivity. (miR-21: microRNA-21; CS: chitosan).

Chemiluminescence and Bioluminescence Imaging for Biosensing and Therapy: In Vitro and In Vivo Perspectives.

Chemiluminescence (CL) and bioluminescence (BL) imaging technologies, which require no external light source so as to avoid the photobleaching, background interference and autoluminescence, have become powerful tools in biochemical analysis and biomedical science with the development of advanced imaging equipment. CL imaging technology has been widely applied to high-throughput detection of a variety of analytes because of its high sensitivity, high efficiency and high signal-to-noise ratio (SNR). Using luciferase and fluorescent proteins as reporters, various BL imaging systems have been developed innovatively for real-time monitoring of diverse molecules in vivo based on the reaction between luciferin and the substrate. Meanwhile, the kinetics of protein interactions even in deep tissues has been studied by BL imaging. In this review, we summarize in vitro and in vivo applications of CL and BL imaging for biosensing and therapy. We first focus on in vitro CL imaging from the view of improving the sensitivity. Then, in vivo CL applications in cells and tissues based on different CL systems are demonstrated. Subsequently, the recent in vitro and in vivo applications of BL imaging are summarized. Finally, we provide the insight into the development trends and future perspectives of CL and BL imaging technologies.

Preparation of a Novel Raman Probe and Its Application in the Detection of Circulating Tumor Cells and Exosomes.

The Raman probe plays an essential role in sensitive surface-enhanced Raman scattering (SERS) assay. Here, a novel Raman probe was developed by assembling gold nanoparticles in triangular pyramid DNA (TP-Au NPs). Such probe with intense electromagnetic hot spots can provide dramatically enhanced Raman scattering. Through assembling recognition DNA on one corner of the TP-DNA, the recognition event is definite and designable. The probe was characterized through TEM, and its SERS superiority was investigated. As models, circulating tumor cells and exosomes were detected with high sensitivity and selectivity by using this probe. Meanwhile, the developed SERS probe can also perform well in real world samples.

Fluorometric determination of mercury(II) by using thymine-thymine mismatches as recognition elements, toehold binding, and enzyme-assisted signal amplification.

A highly sensitive fluorometric method is described for the determination of mercury(II) ions. It is based on (a) the use of a DNA probe containing thymine-thymine mismatches that are employed as Hg(II) recognition elements, (b) subsequent toehold binding, and (c) endocuclease-assisted signal amplification. Target recycling is triggered by exonuclease III. This produces a large amount of ssDNA (defined as primer). Then, the generated primer-initiated strand displacement reaction with the help of polymerase and nicking endonuclease releases the free fluorophore-labelled probe. Under excitation at 532 nm, the fluorescent probe displays emission with a peak at 582 nm. The sensitivity of this method is improved by introduction of nicking endonuclease. The working range of the assay extends from 20 pM to 10 nM, and the detection limit is as low as 6 pM of Hg(II). Graphical abstract Schematic presentation of the fluorometric method for determination of mercury(II). By using a special structure of thymine-thymine mismatches, target-induced toehold binding and enzyme-assisted signal amplification strategy were employed. This method is selective and good performance in real sample application.