Boosting the ammonia synthesis activity of ceria-supported Ru catalysts achieved through trace Pr addition.

The amount of dopant used in conventional cases for improving catalytic performance is higher than 5%. In this work, a strategy to enhance the ammonia synthesis performance of a Ru/CeO2 catalyst by using trace Pr (0.1 mol%) is reported. Owing to the improvement of oxygen defects, Ce3+ concentration and interfaced Ru species, the hydrogen adsorption was enhanced, and the desorption of hydrogen species would be promoted. As a result, Ru/CeO2 with 0.1 mol% Pr shows 1.4 times higher ammonia synthesis rate and excellent stability compared to Ru/CeO2 or the sample with high Pr loading (50 mol% Pr). This study provides a new idea for the design of high-efficiency ammonia synthesis catalysts.

A versatile CRISPR Cas12a-based point-of-care biosensor enabling convenient glucometer readout for ultrasensitive detection of pathogen nucleic acids.

Pathogen nucleic acid detection is of great significance to control the spread of diseases caused by the viruses. Nevertheless, traditional methods for nucleic acid detection such as polymerase chain reaction (PCR) and oligonucleotide microarrays require bulky instruments, which restrain their point-of-care (POC) testing application. Here, we proposed a POC method enabling sensitive detection of pathogen nucleic acids by combining the clustered regularly interspaced short palindromic repeat (CRISPR) Cas12a-based assay and personal glucometer readout (PGM). The quantification of target pathogen DNA by PGM was achieved based on pathogen DNA activates Cas12a ssDNase to cleave magnetic bead-DNA-invertase reporter probe, and separated free invertase to catalyze hydrolysis of sucrose to glucose. Without using nucleic acid amplification technology, we demonstrated here dual signal amplifications based on Cas12a and invertase-mediated catalytic reactions, making it possible to sensitively detect HIV-related DNA or SARS-CoV-2 pseudovirus with the limits of detection of 11.0 fM and 50 copies/µL, respectively. This strategy also showed excellent selectivity as well as potential applicability for detection of HIV in human serum samples or of SARS-CoV-2 in saliva samples. Therefore, our CRISPR-PGM-based dual signal amplifications detection platform might offer a great promise in POC diagnosis of pathogen nucleic acids.

Enhanced ammonia synthesis activity of carbon-supported Mo catalyst by Mo carburization.

It is urgent to develop new efficient ammonia synthesis catalysts using non-precious metals. Herein, the Mo2C species is introduced into a carbon-supported Mo catalyst by in situ carburization of a carbon-supported Mo catalyst in H2. In combination with the presence of the Mo2C phase as well as the enhancement of the graphitization degree of carbon and the amount of the low-valent Mo species, the migration and the exchange of the adsorbed species with the gaseous species are accelerated. As a result, the catalyst with carbonization treatment shows higher ammonia synthesis activity than the sample without carbonization, and the ill effect of the poisoning of reagent gases also is alleviated.

Detection of inflammatory bowel disease (IBD)-associated microRNAs by two color DNA-templated silver nanoclusters fluorescent probes.

Researches demonstrated that circulating miRNAs could be used as novel diagnostic and prognostic potential markers for patients with inflammatory bowel diseases (IBD). It is of great significance in clinical to develop rapid and specific detection methods for miRNAs. Herein, we established a fluorescent probe for ulcerative colitis (UC) activity-associated two serum biomarkers (miR-23a and miR-223) simultaneous detection, which used multi-color fluorescent DNA-stabilized silver nanoclusters (DNA-AgNC) illuminated by a close guanine (G)-rich sequence as a signal transducer and two split DNA probes as recognition units. In principle, the two DNA probe sequences containing AgNC nucleation sequence and G-rich sequence respectively, formed a ternary complex when in the presence of target miRNA through base pairing, so as to induce enhancement of fluorescence emission of AgNC by shortening the distance from G-rich sequence. The combined probes for miR-23a and miR-223 detection generated increased fluorescence signals at 460 nm ex/545 nm em and at 560 nm ex/630 nm em, respectively. With this technique, we successfully quantified the two target miRNAs with high selectivity. Furthermore, the potential clinic applicability of this method was verified by testing the spiked standard miRNAs in human serum. Thus, this one-step, low-cost, and homogenous method offers a great opportunity for disease-associated multiplex miRNAs simultaneous detection.

A novel disease-associated nucleic acid sensing platform based on split DNA-scaffolded sliver nanocluster.

Disease-associated nucleic acids, such as DNAs and miRNAs, are important biomarkers for the diagnosis, prognosis and treatment guidance of human diseases. Therefore, the accurate and sensitive detection of nucleic acid is of great significance for the early diagnosis of diseases. DNA-scaffolded silver nanocluster (DNA-Ag NC) is a new type of probe with good photostability and low toxicity that has been widely used in biomedical analysis. In this work, a new universal sensing platform based on target triggered labeling luminescent DNA-Ag NC for disease-related nucleic acids detection was constructed. The assembled split DNA fragment pair (C4AC4T and C3GT4) could be used as a template to develop a bright green fluorescent Ag NC. According to this phenomenon, we devised two probe sequences DNA 1 and DNA 2, which could hybridize to the same one target and contained a different split fragment of Ag NC' scaffold. The target compelled the split fragments close to each other through base pairing with DNA 1 and DNA 2, thus quantification of the target could be achieved through measuring green fluorescence of Ag NC that produced by assembled scaffold in ternary hybrid products. We applied this platform successfully for miR-362, a potential biomarker of inflammatory bowel diseases (IBD), or HIV-related DNA (hDNA) detection, achieving the detection limits of 6.5 nM and 1.7 nM, respectively. Both of the assays showed excellent reproducibility, selectivity and potential applications in human serum samples. In summary, an economic and convenient universal platform was developed for disease-associated nucleic acid detection.

A novel fluorescent enhancing platform based on DNA-scaffolded silver nanoclusters for potential inflammatory bowel disease-associated microRNA detection.

DNA-scaffolded silver nanoclusters (DNA/AgNC) probes are widely used to detect microRNAs (miRNAs) for diagnosing diseases. However, current available DNA/AgNC probes, which primarily based on fluorescence quenching (turn-off) method, suffer from low detection accuracy caused by bio-matrix interferences. Herein, we designed a new DNA/AgNC-cDNA probe to detect miRNA based on a fluorescence enhancing (turn-on) strategy. Using miR-223, a potential biomarker of inflammatory bowel diseases (IBD), as the target miRNA, we devised the partially hybridized DNA/AgNC-cDNA fluorescent probe. The cDNA was the sequence that completely paired against miR-223 and served as a quencher to the fluorescent DNA/AgNC moiety. Upon the presence of miR-223, which could competitively bind the cDNA, then the DNA/AgNC was set free from the DNA/AgNC-cDNA complex accompanied by an increase in the fluorescence of the DNA/AgNC. Further, by fluorescence decay and polyacrylamide gel electrophoresis (PAGE) experiments, we tentatively addressed the probe working mechanism: the restriction of photo-induced electron-transfer from complementary nucleobases to DNA/AgNC. Compared with the traditional fluorescence turn-off approach, our newly designed probe significantly improved the sensitivity (10 times) and demonstrated excellent specificity. This rapid, label-free, and low-cost fluorescence enhancing method can potentially be applied in the diagnosis of miR-223 associated disease, such as IBD.

Enhanced ammonia synthesis performance of ceria-supported Ru catalysts via introduction of titanium.

The spatial arrangements of Ti species would affect the electronic metal-support interactions and the proportion of Ce3+ sites for ceria-supported Ru catalysts. Ti-Surface-loaded CeO2 supported Ru catalysts exhibited excellent ammonia synthesis activity, which is attributed to a larger proportion of Ru metal, more electrons of Ru species and better adsorption ability of hydrogen and nitrogen.

Hybridization induced fluorescence enhanced DNA-Ag nanocluster/aptamer probe for detection of prostate-specific antigen.

In this work, a label-free Ag nanocluster (AgNC)-based fluorescent probe is proposed to detect tumor marker, prostate-specific antigen (PSA). In the experiments, DNA sequences containing segments complemented to different parts of PSA aptamer were used to synthesize DNA-Ag nanoclusters (DNA-AgNC). Some of the obtained specific DNA-AgNC exhibited significant fluorescence increase after hybridization with PSA aptamer. Based on this, a simple DNA-AgNC/aptamer hybridization probe was fabricated for PSA detection using fluorescence quenching, because competitively specific binding between PSA and its aptamer inhibited the fluorescence enhancement effect of PSA aptamer on DNA-AgNC. The sequence of template DNA, pH and salt concentration of binding buffer, and the concentration of aptamer were optimized. Under optimum conditions, the concentration of PSA within the range of 2-150 ng mL-1 with the detection limit of 1.14 ng mL-1 was detected (3σ; n = 7). This approach was also successfully applied to determine PSA in spiked serum samples. As is well known, this was the first report to realize PSA detection using fluorescent AgNC-based probe. This work would provide reference for construction of AgNC-based probes for detecting other proteins.

Graphene oxide-assisted Au nanoparticle strip biosensor based on GR-5 DNAzyme for rapid lead ion detection.

This study has reported that a GR-5 DNAzyme based lead ion strip biosensor could exhibit an enhanced specificity with the assistance of graphene oxide (GO). This enhancement results from the specific π-stacking interaction between the ribose rings of the nucleobases and the carbon hexagons in GO which can reduce the false positive interference by removing unhybridized ssDNA during the annealing of GR-5 DNAzyme. Meanwhile, conjugate pad was sprayed with two kinds of AuNP-DNA probes, and nitrocellulose membrane test zone and control zone were immobilized with two kinds of biotin-DNA probes, respectively. The limit of detection of this strip biosensor was estimated to be about 0.05 nM (S/N = 3) and 1 nM (with naked eyes) with a linear range from 0.01 to 100 µM. Furthermore, the strip biosensor exhibited excellent selectivity toward Pb2+ in the presence of other divalent metal ions. For real soil samples, the obtained recoveries were in the range from 91.5% to 113.1%.

HIV-related DNA detection through switching on hybridized quenched fluorescent DNA-Ag nanoclusters.

In this paper, DNA containing six cytosines as the formation site for silver nanoclusters (Ag NCs) was adopted as a template for preparing fluorescent DNA-Ag NCs. For the first time, it was found that the fluorescence of DNA-Ag NCs could be quenched after hybridization with their complementary sequence. On the basis of this new phenomenon, we designed a sequence C1 that was completely complementary to human immunodeficiency virus (HIV) DNA, and probe DNA which was partially complementary to C1 for the synthesis of DNA-Ag NCs. The fluorescence of DNA-Ag NCs was quenched after hybridization with C1 and the DNA-Ag NCs/C1 composite was formed, while C1 could be dissociated away from the DNA-Ag NCs by HIV DNA through a strand exchange reaction due to the stronger affinity between HIV DNA and C1, which could switch on the quenched Ag NCs, thus a new "off-on" fluorescence method for HIV DNA detection was developed. In the experiment, the Ag NCs formation site of DNA, the number of base pairs, and the pH and salt concentration of binding buffer were optimized. Under the optimum conditions, the limit of detection for HIV DNA was obtained to be 3.18 nM (3σ/N, n = 7) with the linear range of 15-150 nM for the 150 nM DNA-Ag NCs/C1 probe. Besides, the probe showed excellent specificity to HIV DNA, and even distinguished one nucleotide mismatched HIV DNA.

Nitrogen-doped graphene quantum dot for direct fluorescence detection of Al3+ in aqueous media and living cells.

Graphene quantum dot (GQD) has been attractive in analytical science field due to its low toxicity, stable photoluminescence. Herein, nitrogen-doped GQD (N-GQD) was prepared by a facile solvothermal treatment of GO using dimethylformamide, and exhibited a green emission with 23.1% quantum yield. The N-GQD probe showed a selective and sensitive fluorescence enhancement response to Al3+, the mechanism might be the formation of a complex between Al3+ and N-GQD constrained the photo-induced electron transfer (PET) process of N-GQD itself. With Benesi-Hildebrand equation, the binding constant and molar ratio between N-GQD and Al3+ was calculated to be 4.6 × 104Lmol-1 and 1:1 respectively. The pKa value of N-GQD was also determined to be 4.4 by capillary electrophoresis. In pH 4.0 PBS solution, there was a good linear relation between the fluorescence intensity and the logarithm of concentration of Al3+ in the range of 2.5-75µmolL-1, the limit of detection (3σ) was 1.3µmolL-1. This "Off - On" fluorescence method had been applied to accurate quantification of aluminum in hydrotalcite tablets. What's more, the fluorescence switch property of N-GQD was explored by alternate addition of Al3+ and EDTA. The probe was also utilized for detection Al3+ in living cells due to its excellent biocompatibility.

Visual detection of trace lead ion based on aptamer and silver staining nano-metal composite.

In this paper, visual detection of trace lead ion was established by aptamer and silver staining. The basic strategy was that aminated PS2.M aptamer was immobilized onto slide and formed stable G-quadruplex structure. PbS was generated by adding S2-, and it catalyzed subsequent silver staining reaction, through the silver staining amplification effect, the slide presented visible ash black. The gray value of slide after silver staining was analyzed and the semi-quantitative detection of Pb2+ in solution was realized. The results showed that optical darkness ratio (ODR) and logarithmic value of Pb2+ concentration had a good linear relationship (R2 = 0.951) over the range of 0.5-10 µM. In addition, there was no obvious interference of other common metal ions for the detection, indicating that this method presented outstanding selectivity. And it was also used for qualitative and semi-quantitative determination of Pb2+ in soil sample successfully.

In Vivo Computed Tomography/Photoacoustic Imaging and NIR-Triggered Chemo-Photothermal Combined Therapy Based on a Gold Nanostar-, Mesoporous Silica-, and Thermosensitive Liposome-Composited Nanoprobe.

Safe multifunctional nanoplatforms that have multiple therapeutic functions integrated with imaging capabilities are highly desired for biomedical applications. In this paper, targeted chemo-photothermal synergistic therapy and photoacoustic/computed tomography imaging of tumors were achieved by one novel multifunctional nanoprobe (GMS/DOX@SLB-FA); it was composed of a gold nanostar core and a doxorubicin (DOX)-loaded mesoporous silica shell (GMS), which was coated with a folic acid (FA)-modified thermosensitively supported lipid bilayer (SLB-FA) as a gatekeeper. The multifunctional probe had perfect dispersion and stability; 2.1 nm mesoporous pores and 208 nm hydration particle sizes were obtained. In vitro studies indicated that the drug-loaded probe had excellent ability to control the release of DOX, with 71.98 ± 2.52% cumulative release after laser irradiation, which was significantly higher than that of unirradiated control group. A survival rate of 72.75 ± 4.37% of HeLa cells at 57.75 µg/mL probe also demonstrated the low cytotoxicity of the targeted probe. Both in vitro and in vivo results showed that the probe could achieve targeted photoacoustic imaging of tumors because of the fact that the FA-modified probe could specifically recognize the overexpressed FA receptors on tumor cells; meanwhile, the probe could also achieve the chemo-photothermal synergistic therapy of tumors through controlling the drug release from mesoporous channels by a near-infrared laser. Therefore, the probe had great potential in the early diagnosis and treatment of cancer.

Detection of adenosine triphosphate in HeLa cell using capillary electrophoresis-laser induced fluorescence detection based on aptamer and graphene oxide.

A method for ATP quantification based on dye-labeled aptamer/graphene oxide (aptamer/GO) using capillary electrophoresis-laser induced fluorescence (CE-LIF) detecting technique has been established. In this method, the carboxyfluorescein (FAM)-labelled ATP aptamers were adsorbed onto the surface of GO, leading to the fluorescence quenching of FAM; after the incubation with a limited amount of ATP, stronger affinity between ATP aptamer and ATP resulted in the desorption of aptamers and the fluorescence restoration of FAM. Then, aptamer-ATP complex and excess of aptamer/GO and GO were separated and quantified by CE-LIF detection. It was shown that a linear relation was existing in the CE-LIF peak intensity of aptamer-ATP and ATP concentration in range of 10-700 µM, the regression equation was F=1.50+0.0470C(ATP) (R(2)=0.990), and the limit of detection was 1.28 µM (3S/N, n=5), which was one order magnitude lower than that of detection in solution by fluorescence method. The approach with excellent specificity and reproducibility has been successfully applied to detecting concentration of ATP in HeLa cell.

An engineered coiled-coil polypeptide assembled onto quantum dots for targeted cell imaging.

Quantum dot (QD)-polypeptide probes have been developed through the specific metal-affinity interaction between polypeptides appended with N-terminal polyhistidine sequences and CdSe/ZnS core-shell QDs. The size and charge of a QD-polypeptide can be tuned by using different coiled-coil polypeptides. Compared to glutathione-capped QDs (QD-GSH), QD-polypeptide probes showed an approximately two- to three-fold luminescence increase, and the luminescence increase was not obviously related to the charge of the polypeptide. QD-polypeptide probes with different charge have a great effect on nonspecific cellular uptake. QD-polypeptide probes with negative charge exhibited lower nonspecific cellular uptake in comparison to the QD-GSH, while positively charged QD-polypeptide probes presented higher cellular uptake than the QD-GSH. A targeted QD-ARGD probe can obviously increase targeted cellular uptake in α v ß 3 overexpressing HeLa cells compared to QD-A. In addition, QD-polypeptide probes showed lower in vitro cytotoxicity compared to the original QDs. These results demonstrate that these QD-polypeptide probes with high specific cellular uptake, high fluorescence intensity and low background noise are expected to have great potential applications in targeted cell imaging.

Multifunctional magnetic-hollow gold nanospheres for bimodal cancer cell imaging and photothermal therapy.

Multifunctional nanocomposites combining imaging and therapeutic functions have great potential for cancer diagnosis and therapy. In this work, we developed a novel theranostic agent based on hollow gold nanospheres (HGNs) and superparamagnetic iron oxide nanoparticles (SPIO). Taking advantage of the excellent magnetic properties of SPIO and strong near-infrared (NIR) absorption property of HGNs, such nanocomposites were applied to targeted magnetic resonance imaging (MRI) and photoacoustic imaging (PAI) of cancer cells. In vitro results demonstrated they displayed significant contrast enhancement for T2-weighted MRI and strong PAI signal enhancement. Simultaneously, the nanocomposites exhibited a high photothermal effect under the irradiation of the near-infrared laser and can be used as efficient photothermal therapy (PTT) agents for selective killing of cancer cells. All these results indicated that such nanocomposites combined with MRI-PAI and PTT functionality can have great potential for effective cancer diagnosis and therapy.

Capillary electrophoresis-chemiluminescence detection for carcino-embryonic antigen based on aptamer/graphene oxide structure.

A new strategy is proposed for determination of carcino-embryonic antigen (CEA) based on aptamer/graphene oxide (Apt/GO) by capillary electrophoresis-chemiluminescence (CE-CL) detection system. CEA aptamer conjugated with horseradish peroxidase (HRP) firstly mixes with GO, and the CL will be quenched because the stack of HRP-Apt on GO leads to chemiluminescence resonance energy transfer (CRET). When CEA exists, the specific combination of HRP-Apt and CEA can form HRP-Apt-CEA complex, which dissociates from GO. Then, the CL catalyzed by HRP-Apt-CEA complex can be detected without any CRET, and the content of CEA can be estimated by the CL intensity. It has been proved that the interference issue resulted from free HRP-Apt is solved well by mixing GO firstly with HRP-Apt, which blocks the free HRP-Apt's CL signal due to CL quenching effect of GO; and the interference resulted from GO to CL is also solved by CE, then the sensitivity and accuracy can be greatly improved. Results also showed that the CL intensity had a linear relationship with the concentration of CEA in the range from 0.0654 to 6.54 ng/mL, and the limit of detection was approximately 4.8 pg/mL (S/N = 3). This proposed method with high specificity offers a new way for separation and determination of biomolecule, and has good potential in application of biochemistry and bioanalysis.

High transfection efficiency of quantum dot-antisense oligonucleotide nanoparticles in cancer cells through dual-receptor synergistic targeting.

Incorporating ligands with nanoparticle-based carriers for specific delivery of therapeutic nucleic acids (such as antisense oligonucleotides and siRNA) to tumor sites is a promising approach in anti-cancer strategies. However, nanoparticle-based carriers remain insufficient in terms of the selectivity and transfection efficiency. In this paper, we designed a dual receptor-targeted QDs gene carrier QD-(AS-ODN+GE11+c(RGDfK)) which could increase the cellular uptake efficiency and further enhance the transfection efficiency. Here, the targeting ligands used were peptides GE11 and c(RGDfK) which could recognize epidermal growth factor receptors (EGFR) and integrin ανß3 receptors, respectively. Quantitative flow cytometry and ICP/MS showed that the synergistic effect between EGFR and integrin ανß3 increased the cellular uptake of QDs carriers. The effects of inhibition agents showed the endocytosis pathway of QD-(AS-ODN+GE11+c(RGDfK)) probe was mainly clathrin-mediated. Western blot confirmed that QD-(AS-ODN+GE11+c(RGDfK)) could further enhance gene silencing efficiency compared to QD-(AS-ODN+GE11) and QD-(AS-ODN+c(RGDfK)), suggesting this dual receptor-targeted gene carrier achieved desired transfection efficiency. In this gene delivery system, QDs could not only be used as a gene vehicle but also as fluorescence probe, allowing for localization and tracking during the delivery process. This transport model is very well referenced for non-viral gene carriers to enhance the targeting ability and transfection efficiency.

Highly sensitive and selective determination of cupric ions by using N,N'-bis(salicylidene)-o-phenylenediamine as fluorescent chemosensor and related applications.

A sensitive and selective copper(II) fluorescence Schiff base chemical sensor receptor 1 (short for N,N'-bis(salicylidene)-o-phenylenediamine) has been prepared. The fluorescence of receptor 1 in pH 8.2 phosphate buffer solution can be dramatically quenched by Cu(2+), whereas it is nearly unaffected by other metal ions. Based on this, a sensitive and selective fluorescent quenching method for Cu(2+) detection has been established. Under the optimum conditions, a good linear relation exists between the quenching efficiency (F0/F) and the concentration of Cu(2+) in the range of 1.0 × 10(-7)-2.5 × 10(-6) mol L(-1). The detection limit (3σ) for Cu(2+) determination is 2.0 × 10(-8) mol L(-1). The present method has been successfully used for quantification of Cu(2+) in soybean milk powder. Furthermore, the fluorescence switch property of the system was explored, and the system might be applied for determination of glutathione and construction of molecular logic gate.

Determination of ethylenediaminetetraacetic acid base on the reversion of fluorescence quenching of 2-pyridinecarbaldehyde-p-phenyldihydrazone by ferric iron.

A fast and sensitive fluorometric method for determination of an emerging pollutant, ethylenediaminetetraacetic acid (EDTA), was proposed based on the reversion of fluorescence quenching of 2-pyridinecarbaldehyde-p-phenyldihydrazone by ferric iron. It was found that Fe(3+) could selectively quench the fluorescence of 2PC-PPH, while EDTA could recover it due to its strong coordination ability with Fe(3+). Based on this, we established an "Off and On" fluorescence method for determination of EDTA. Under the optimum conditions, the linear range and detection limit (3S/N) for EDTA determination were 2.0 × 10(-6)~1.4 × 10(-5) mol L(-1) and 1.5 × 10(-7) mol L(-1), respectively. The relative standard deviation for the determination of 7.0 × 10(-6)mol L(-1) EDTA was 3.7% (n=11). The method was successfully applied to determination of EDTA in tap water, lake water and investigating degradation of EDTA by TiO2/UV process.