Programmable Tetrahedral DNA-RNA Nanocages Woven with Stimuli-Responsive siRNA for Enhancing Therapeutic Efficacy of Multidrug-Resistant Tumors.

Multidrug resistance (MDR) is a major obstacle limiting the effectiveness of chemotherapy against cancer. The combination strategy of chemotherapeutic agents and siRNA targeting drug efflux has emerged as an effective cancer treatment to overcome MDR. Herein, stimuli-responsive programmable tetrahedral DNA-RNA nanocages (TDRN) have been rationally designed and developed for dynamic co-delivery of the chemotherapeutic drug doxorubicin and P-glycoprotein (P-gp) siRNA. Specifically, the sense and antisense strand sequences of the P-gp siRNA, which are programmable bricks with terminal disulfide bond conjugation, are precisely embedded in one edge of the DNA tetrahedron. TDRN provides a stimuli-responsive release element for dynamic control of functional cargo P-gp siRNA that is significantly more stable than the "tail-like" TDN nanostructures. The stable and highly rigid 3D nanostructure of the siRNA-organized TDRN nanocages demonstrated a notable improvement in the stability of RNase A and mouse serum, as well as long-term storage stability for up to 4 weeks, as evidenced by this study. These biocompatible and multifunctional TDRN nanocarriers with gold nanocluster-assisted delivery (TDRN@Dox@AuNCp) are successfully used to achieve synergistic RNAi/Chemo-therapy in vitro and in vivo. This programmable TDRN drug delivery system, which integrates RNAi therapy and chemotherapy, offers a promising approach for treating multidrug-resistant tumors.

Electrochemical Detection of Viral Nucleic Acids by DNA Nanolock-Based Porous Electrode Device.

Developing rapid, sensitive, and facile nucleic acid detection technologies is of paramount importance for preventing and controlling infectious diseases. Benefiting from the advantages such as rapid response, low cost, and simple operation, electrochemical impedance spectroscopy holds great promise for point-of-care nucleic acid detection. However, the sensitivity of electrochemical impedance spectroscopy for low molecular weight nucleic acids testing is still limited. This work presents a DNA nanolock-based porous electrode to improve the sensitivity of electrochemical impedance spectroscopy. Once the target nucleic acids are recognized by the DNA probes, the pore-attached DNA nanolock caused remarkable impedance amplification by blocking the nanopores. Taking SARS-CoV-2 nucleic acid as a model analyte, the detection limit of the porous electrode was as low as 0.03 fM for both SARS-CoV-2 RNA and DNA. The integration of a porous electrode with a wireless communicating unit generates a portable detection device that could be applied to direct SARS-CoV-2 nucleic acid testing in saliva samples. The portable device could effectively distinguish the COVID-19 positive and negative samples, showing a sensitivity of 100% and a specificity of 93%. Owing to its rapid, ultrasensitive, specific, and portable features, the as-designed DNA nanolock and porous electrode-based portable device holds great promise as a point-of-care platform for real-time screening of COVID-19 and other epidemics.

Homogeneous Electrochemical Aptasensor for Sensitive Detection of Zearalenone Using Nanocomposite Probe and Silica Nanochannel Film.

Developing rapid and efficient analytical methods is of great importance for food safety Herein, we present a novel homogeneous electrochemical aptasensor for ultrasensitive quantitative determination of zearalenone (ZEN) based on a nanocomposite probe and silica nanochannel film. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and UV-Vis characterization techniques confirm that graphene oxide (GO) bears an aromatic conjugated structure, along with hydroxyl and carboxyl groups, facilitating the subsequent adsorption of cationic redox hexa-ammine-ruthenium (III) (Ru(NH3)63+) and anionic ZEN aptamer, to form a Ru(NH3)63+-ZEN aptamer-GO nanocomposite probe in a homogeneous solution. Vertically-ordered mesoporous silica films (VMSF) bearing silanol groups can be simply grown on the solid indium tin oxide (ITO) electrode surface and enable the selective preconcentration of Ru(NH3)63+, eventually leading to signal amplification. Since the detachment of Ru(NH3)63+ from the GO surface by the recognized ZEN aptamer in the presence of ZEN, more free Ru(NH3)63+ is released in solution and produces enhanced redox signals at the VMSF modified ITO electrode, allowing quantitative detection of ZEN. On the basis of the above sensing strategy, the proposed homogeneity, due to the assistance of graphene, as well as of the signal amplification and anti-fouling effects of VMSF, accurate analysis of ZEN can be realized in maize and Chinese chestnut samples.

Energy Level Engineering in Gold Nanoclusters for Exceptionally Bright NIR Electrochemiluminescence at a Low Trigger Potential.

Electrochemiluminescence (ECL) is a widely used light output mechanism from electrochemical excitation. Understanding the intrinsic essence for ideal ECL generation remains a fundamental challenge. Here, based on the molecular orbital theory, we reported an energy level engineering strategy to regulate the ECL performance by using ligand-protected gold nanoclusters (AuNCs) as luminophores and N,N-diisopropylethylamine (DIPEA) as a coreactant. The energy level matching between the AuNCs and DIPEA effectively promoted their electron transfer reactions, thus improving the excitation efficiency and reducing the trigger potential. Simultaneously, the narrow band gap of the AuNCs further enabled enhanced emission efficiency. Using the energy level engineering theory developed here, a dual-enhanced strategy was proposed, and ß-CD-AuNCs were designed to further verify this mechanism. The ß-CD-AuNCs/DIPEA system resulted in highly stable near-infrared ECL with an unprecedented ECL efficiency (145-fold higher than that of the classic Ru(bpy)32+/tetra-n-butylammonium perchlorate system) and a low trigger potential of 0.48 V. A visual NIR-ECL based on this ECL system was successfully realized by an infrared camera. This work provides an original mechanistic understanding for designing efficient ECL systems, which promises to be a harbinger for broad applicability of this strategy for other ECL systems and ECL sensing platforms.

Isobutyric acid promotes colorectal cancer metastasis through activating RACK1.

Colorectal cancer (CRC) metastasis plays a crucial role in disease progression, yet the regulatory mechanisms underlying metastasis remain incompletely understood. Isobutyric acid (IBA), a short-chain fatty acid found at high levels in serum of CRC patients, has been shown to be a critical metabolite influencing CRC proliferation. However, its role in tumor metastasis remains unknown. Here, utilizing liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis, we found that levels of IBA were significantly higher in patients with distant organ metastasis of CRC than in those without. Furthermore, IBA promoted CRC metastasis both in vitro and in vivo. Mass spectrometry, immunofluorescence, and cellular thermal shift assay revealed that IBA interacts with RACK1. Mechanistically, IBA binding to and activating RACK1 promotes regulation of downstream Akt and FAK signaling and CRC metastasis. Collectively, our study highlights the critical interplay between IBA and RACK1 and its impact on tumor metastasis. This study suggests that targeting the IBA-RACK1 signaling axis may be an effective therapeutic strategy for controlling CRC metastasis.

In Situ Visualization of Epidermal Growth Factor Receptor Nuclear Translocation with Circular Bivalent Aptamer.

Epidermal growth factor receptor (EGFR) nuclear translocation correlates with the abnormal proliferation, migration, and anti-apoptosis of tumor cells. Monitoring EGFR nuclear translocation provides insights into the molecular mechanisms underlying cancers. EGFR nuclear translocation includes two processes, EGFR phosphorylation and phosphorylated EGFR translocation to the nucleus. With the help of aptamers, probes that can achieve the first step of anchoring phosphorylated EGFR have been developed. However, the EGFR nuclear translocation can last for hours, posing a challenge to monitor the entire nuclear translocation in living cells. Herein, we designed a circular bivalent aptamer-functionalized optical probe with greatly enhanced stability for long-term visualization of EGFR nuclear translocation in situ. The results of cell experiments show that the probe could monitor the entire nuclear translocation of EGFR. The findings of tissue and in vivo experiments demonstrate that the probe can evaluate the development and progression of tumors by imaging EGFR nuclear translocation in situ. The proposed approach allows us to monitor EGFR nuclear translocation in the long term, indicating its great potential in investigating the mechanisms of cancers and guiding for tumor treatment.

Gold Nanocluster Probe-Based Electron-Transfer-Mediated Electrochemiluminescence Sensing Strategy for an Ultrasensitive Copper Ion Detection.

Exploration of a novel and efficient sensing mechanism of Au nanocluster (AuNC)-based electrochemiluminescence (ECL) sensors is still a great challenge and opportunity for further applications. Herein, we proposed that the electron transfer (ET) could be used as a novel sensing regulation factor for the construction of an ECL-sensing platform based on the AuNC probe. As a proof-of-concept, the ECL quenching effect and mechanism of Cu2+ on pre-oxidation-treated l-methionine-capped AuNCs (OM-AuNCs) was investigated in detail. The results revealed that after the electrochemical excitation of the AuNC probe, the electron is transferred from the highest occupied molecular orbital (HOMO) of Met-Cu2+ to that of the OM-AuNCs, along with the ET from lowest-unoccupied molecular orbital (LUMO) of the OM-AuNCs back to the HOMO of Met-Cu2+, leading to the ECL quenching of OM-AuNCs. Since the ECL intensity of OM-AuNCs is sensitively affected by the ET process, a preferable linear dependence was obtained in the concentration range from 1.0 × 10-18 to 1.0 × 10-14 M with high selectivity. More importantly, a record low detection limit (LOD, 2.3 × 10-20 M) at the single copper ion level has been realized without any other amplification technique. Furthermore, the actual sample detection for Cu2+ exhibited satisfactory results. Therefore, this study enriches an ET-mediated ECL application and promotes a more rational design of ECL sensors.

Co-Reactant-Mediated Low-Potential Anodic Electrochemiluminescence Platform and Its Immunosensing Application.

Screening high-performance anodic electrochemiluminescence (ECL) systems with low triggering potential is a promising way to broaden their applications. In addition to electrochemiluminophore, co-reactant also plays an important role in the ECL process, since the oxidation of co-reactants is one of the most important steps in the anodic ECL process. Herein, a novel co-reactant-mediated high-performance low-potential Au nanocluster (AuNC)-based ECL system has been successfully developed. Benefiting from the isopropyl substitution and hydroxyl addition to the triethylamine (TEA), the BSA-AuNC/2-(diisopropylamino)ethanol (DIPEA-OH) ECL system achieved higher energy efficiency at a lower potential of 0.75 V. In addition, compared with the BSA-AuNC/TEA system, the ECL intensity and quantum yield (ΦECL) with DIPEA-OH as a co-reactant increased 22.34-fold and 13-fold (as high as 68.17%), respectively. Based on the low potential, high ΦECL of the AuNC/DIPEA-OH ECL system, a sandwich-type immunosensor has been constructed for a highly selective SARS-CoV-2 N protein assay. In the absence of any complex signal amplification strategies, the ECL immunosensor for the SARS-CoV-2 N protein detection showed a linear range of 0.001-100 ng/mL and a detection limit of 0.35 pg/mL. Moreover, the ECL platform had good reproducibility and stability and exhibited acceptable detection performance in the detection of actual serum samples. This work established a framework for in-depth design and study of anode ECL co-reactants for AuNCs and other luminophores, and expanded the potential application of ECL sensors in the clinical diagnosis of COVID-19.

Facile high-quantum-yield sulfur-quantum-dot-based photoluminescent probe for nifedipine detection.

Monitoring of dihydropyridine drugs, such as nifedipine (NIF), has attracted considerable attention owing to the side effects arising from the consumption of such drugs. Herein, a highly sensitive and facile fluorescence-sensing platform based on a high-quantum-yield sulfur quantum dot (SQDs) probe for NIF detection is proposed. Based on the principle of the inner filter effect, the rapid detection of NIF with high sensitivity is successfully realized on the basis of the change in the fluorescence signal due to the quenching effect of NIF on SQDs. The results show a good linear relationship between the NIF concentration and fluorescence intensity within the range of 5-150 µmol/L, with a low detection limit of 1.63 µmol/L (S/N = 3). Moreover, because no surface modification or establishment of any coupling between the receptor and the fluorophore is necessary, this approach provides considerable flexibility and simplicity for the construction of a fluorescence sensor and substantially reduces the detection time. A systematic investigation was conducted to verify the applicability of this method for the analysis of pharmaceutical components in NIF tablets. This study not only promotes the design and development of a fluorescence analysis platform for NIF detection, but also facilitates the fabrication of novel SQD-based fluorescence-sensing systems for the molecular detection of drugs. Proposal for a facile nifedipine assay method based on the inner filter effect of nifedipine to high-quantum-yield sulfur quantum dots, and realizing nifedipine detection in tablets and human urine samples.

6-Aza-2-thio-thymine-gold nanoclusters: an excellent candidate in the photoelectrochemical field.

The high performance of the photoelectrochemical (PEC) properties of AuNCs can be achieved with 6-aza-2-thio-thymine-AuNCs (ATT-AuNCs) as a photoactive material. The ATT-AuNCs yielded a cathodic photocurrent density as high as 88 µA cm-2 with O2 as electron acceptor, which is three orders of magnitude higher than those of other AuNCs in aqueous solutions. Moreover, ATT-AuNCs also show a higher carrier density, shorter Debye length, and smaller depletion layer width than those of reported AuNCs. This work not only reveals the PEC performance and mechanism of ATT-AuNCs, but also establishes a framework for in-depth design and studying the PEC performance of AuNCs.

Ultrasensitive Glutathione-Mediated Facile Split-Type Electrochemiluminescence Nanoswitch Sensing Platform.

Seeking for an advanced electrochemiluminescence (ECL) platform is still an active and continuous theme in the ECL-sensing realm. This work outlines a femtomolar-level and highly selective glutathione (GSH) and adenosine triphosphate (ATP) ECL assay strategy using a facile split-type gold nanocluster (AuNC) probe-based ECL platform. The system utilizes GSH as an efficient etching agent to turn on the MnO2/AuNC-based ECL nanoswitch platform. This method successfully achieves an ultrasensitive detection of GSH, which significantly outperformed other sensors. Based on the above excellent results, GSH-related biological assays have been further established by taking ATP as a model. Combined with the high catalytic oxidation ability of DNAzyme, this ECL sensor can realize ATP assay as low as 1.4 fmol without other complicated exonuclease amplification strategies. Thus, we successfully achieved an ultrahigh sensitivity, extremely wide dynamic range, great simplicity, and strong anti-interference detection of ATP. In addition, the actual sample detection for GSH and ATP exhibits satisfactory results. We believe that our proposed high-performance platform will provide more possibilities for the detection of other GSH-related substances and show great prospect in disease diagnosis and biochemical research.

Electrochemiluminescence Immunoassay Platform with Immunoglobulin G-Encapsulated Gold Nanoclusters as a "Two-In-One" Probe.

Biomolecule-functionalized Au nanoclusters (AuNCs) have drawn great interest in the electrochemiluminescence (ECL) field due to their unique optical/electrical properties, biocompatibility, and versatile bioapplication potentials. Herein, we proposed a two-in-one ECL probe of immunoglobulin G-encapsulated AuNCs (IgG-AuNCs) for the development of a high-performance ECL immunoassay (ECLIA) platform. The IgG-AuNCs were not only used as an ECL probe due to their excellent anodic ECL performance with triethylamine (TEA) as a coreactant but also used as the biorecognition element because of their well-retained bioactivity of the IgG. As a proof of concept, a new type of competitive immunosensing platform has been applied to detect IgG representing several merits of facile preparation, rapid detection, sample saving, and good analytical performance. The sensing platform exhibited a linear range of 0.5-50,000 ng/mL with a limit of detection of 0.06 ng/mL for IgG detection with high selectivity. In addition, this convenient ECLIA platform also performed well in real serum sample detection. Notably, our work not only proved the "two-in-one" immuno-AuNC probe-based ECLIA strategy but also developed a rational framework for study of ECL bioassay platforms based on multifunctional AuNCs and other related nanomaterials.

Regulating Valence States of Gold Nanocluster as a New Strategy for the Ultrasensitive Electrochemiluminescence Detection of Kanamycin.

Monitoring of kanamycin residue has attracted considerable attention owing to the potential harm caused by the abuse of kanamycin. However, the detection of kanamycin has been limited owing to its electrochemical and optical inertness. Herein, we report a facile and highly efficient electrochemiluminescence (ECL) strategy for the detection of kanamycin based on the valence state effect of gold nanocluster (AuNC) probes. It is proven that Au0 in chemically reduced AuNCs (CR-AuNCs) could be oxidized to AuI via the redox reaction between kanamycin and CR-AuNCs in the presence of H2O2, resulting in ECL quenching due to the valence state change of CR-AuNCs. Because the ECL of the AuNC probes is sensitively affected by the valence state, excellent sensitivity for kanamycin was achieved without any signal amplification operation and aptamers. A preferable linear-dependent curve was acquired in the detection range from 1.0 × 10-11 to 3.3 × 10-5 M with an extremely low detection limit of 1.5 × 10-12 M. The proposed kanamycin sensing platform is very simple and shows high selectivity and an extremely broad linear range detection of kanamycin. Furthermore, the proposed sensing platform can detect kanamycin in milk samples with excellent recoveries. Therefore, this sensing strategy provides an effective and facile way to detect kanamycin and can help promote the understanding of the constructed mechanism of the AuNC-based ECL system, thus greatly broadening its potential application in ECL fields.

Split-type electrochemiluminescent gene assay platform based on gold nanocluster probe for human papillomavirus diagnosis.

Persistent high-risk human papillomavirus (HPV) infection is the leading cause of cervical cancer. Efficient detection of HPV16 E7 is necessary for early diagnosis and cure of the disease. Here, a novel and high-performance Au nanocluster (AuNC) probe-based split-type electrochemiluminescent (ECL) assay platform has been established to detect these oncogenes, in which the nucleic acid hybridization assay and the ECL measurements are performed independently. The proposed approach combines superior magnetic nanobead enrichment and separation technology, specific nucleic acid hybridization technology, and high-efficiency AuNC probe ECL strategy, and shows excellent advantages. First, the split-type ECL sensing platform can effectively avoid interference from biological samples and adequately uses the ECL efficiency of the AuNC probe. Furthermore, the ultrahigh sensitivity assay of HPV DNA can be achieved without any complex nucleic acid amplification technique. Taking advantage of the above merits of split-type detection, the ECL DNA sensor achieved ideal low detection of 6.8 aM and a wide dynamic range bridging 10 orders of magnitude HPV16 E7. Furthermore, together with its favorable and powerful specificity, high sensitivity, and good selectivity, this strategy could detect HPV16 E7 DNA in human samples, which showed great consistency with the FDA-approved approach (Hybrid capture 2, HC2). Therefore, this work proposes a facile and reliable split-type ECL platform for HPV diagnosis and shows great potential for the early diagnosis of other diseases.

Size-focusing results in highly photoluminescent sulfur quantum dots with a stable emission wavelength.

Sulfur quantum dots (SQDs) are a new kind of functional nanomaterial, but several challenges still exist in relation to their synthesis and application, such as low-yield and time-consuming synthetic methods, low photoluminescence quantum yields (PLQYs), and the non-selectivity of their detection mechanisms. Herein, we report the drastic enhancement of the fluorescence performance of water-soluble SQDs via the one-pot synthesis of size-focusing QDs using ultrasound microwave radiation. The synthetic period has been greatly shortened to 2 h via the present process. Notably, the proposed SQDs exhibit a highly stable emission wavelength with a record high PLQY of 58.6%. The mechanistic study indicates that size-focusing is a key factor relating to the proposed high-performance SQDs. As they also have robust stability, the proposed SQDs show a wide range of potential applications. Inspired by the characteristic properties of the SQDs and specific analytes, a simple SQD-based fluorescence sensing platform, via a redox-reaction-mediated mechanism, has been successfully developed for the rapid and selective detection of Ce(iv). In addition, this system has been effectively applied to some Ce(iv)-related biological assays, such as ascorbic acid (AA) analysis. This work is an important breakthrough in the SQD field, opening up avenues for solving the challenging problems relating to SQD-based probes, enriching the fundamental understanding of them, and greatly extending their applications, especially in biomedicine.

Mechanistic Insight into a Novel Ultrasensitive Nicotine Assay Base on High-Efficiency Quenching of Gold Nanocluster Cathodic Electrochemiluminescence.

Monitoring nicotine concentrations in human fluids is extremely crucial owing to the harmful effect of nicotine on human health. Herein, it is shown that nicotine could quench the cathodic electrochemiluminescence (ECL) of gold nanoclusters (AuNCs) with high efficiency. The ECL quenching mechanism of nicotine was studied in detail using various experimental tools and theoretical calculations. It was concluded that the strongly oxidizing intermediate SO4•-, produced from K2S2O8, could oxidized nicotine, resulting in ECL emission quenching. On the basis of this high-efficiency ECL quenching of the AuNCs/K2S2O8 system, a recyclable, ultrasensitive, and selective ECL sensing platform for nicotine detection was proposed. Even in the absence of any complex signal amplification techniques, the ECL sensor for nicotine detection showed an unprecedentedly low detection limit of 7.0 × 10-13 M (S/N = 3) and a wide linear range over 8 orders of magnitude. Most remarkably, it could be successfully used for nicotine detection in human urine samples. This is expected to promote the investigations and applications on nicotine-related diseases. We believe that the proposed ECL platform can hold great prospects for commercialization in biomedical fields and tobacco industries.

6-Aza-2-Thio-Thymine Stabilized Gold Nanoclusters as Photoluminescent Probe for Protein Detection.

This study puts forward an efficient method for protein detection in virtue of the tremendous fluorescence enhancement property of 6-aza-2-thio-thymine protected gold nanoclusters (ATT-AuNCs). In-depth studies of the protein-induced photoluminescence enhancement mechanism illustrate the mechanism of the interaction between ATT-AuNCs and protein. This new-established probe enables feasible and sensitive quantification of the concentrations of total protein in real samples, such as human serum, human plasma, milk, and cell extracts. The results of this proposed method are in good agreement with those determined by the classical bicinchoninic acid method (BCA method).

Dual Enhancement of Gold Nanocluster Electrochemiluminescence: Electrocatalytic Excitation and Aggregation-Induced Emission.

Ligand-protected gold nanoclusters (AuNCs) have emerged as a new class of electrochemiluminescence (ECL) luminophores for their interesting catalytic and emission properties, although their quantum yield (ΦECL ) in aqueous medium is low with a poor mechanistic understanding of the ECL process. Now it is shown that drying AuNCs on electrodes enabled both enhanced electrochemical excitation by an electrocatalytic effect, and enhanced emission by aggregation-induced ECL (AIECL) for 6-aza-2-thiothymine (ATT) protected AuNCs with triethylamine (TEA) as a coreactant. The dried ATT-AuNCs/TEA system resulted in highly stable visual ECL with a ΦECL of 78 %, and a similar enhancement was also achieved with methionine-capped AuNCs. The drying enabled dual-enhancement mechanism has solved a challenging mechanistic problem for AuNC ECL probes, and can guide further rational design of ECL emitters.

Versatile High-Performance Electrochemiluminescence ELISA Platform Based on a Gold Nanocluster Probe.

This report outlines a versatile high-performance electrochemiluminescence (ECL) enzyme-linked immunosorbent assay (ELISA) platform, which combines the merits of high-quantum-yield Au nanocluster (AuNC) probe-based ECL technology, the efficient ECL-resonance energy-transfer (ECL-RET) strategy, and highly sensitive and specific ELISA technology. The ECL detection procedure was performed on a recyclable MnO2/AuNC-modified glassy carbon electrode interface by taking advantage of the ECL-RET between the AuNC probe and MnO2 nanomaterials (NMs) to quench the ECL intensity. The etching of MnO2 NMs by the product of ALP-based ELISA recovers the ECL signal. Notably, the ELISA process and the ECL detection procedure in this system are independent. Thus, the ECL-ELISA system can effectively avoid the influence of complex biological samples, and the ECL efficiency of the AuNC probe can be used readily. As demonstrated on TNF-α, because of the abovementioned characteristics, the ECL-ELISA platform presented an extremely wide dynamic range, with a detection limit of 2 orders lower than ELISA. Moreover, the system was also applicable for ultrahigh sensitive detection of various disease-related proteins and able to detect trace biomarkers in real serum samples. Therefore, this multifunctional ECL assay platform is versatile, facile, ultrasensitive, recyclable, and sufficiently straightforward for trace biomarker detection in complex biological samples. This approach not only enriches the foundational study of ECL devices but also greatly expands the potential application of ECL sensors in biological testing and clinical high-throughput diagnosis.