Sensitive electrochemical flow injection analysis of H2O2 released from cells with a pass-through mode.

Conventional plate electrodes were commonly used in electrochemical flow injection analysis and only part of molecules diffused to the plane of electrodes could be detected, which would limit the performance of electrochemical detection. In this study, a low-cost native stainless steel wire mesh (SSWM) electrode was integrated into a 3D-printed device for electrochemical flow injection analysis with a pass-through mode, which is different compared with previous flow-through mode. This strategy was applied for sensitive analysis of hydrogen peroxide (H2O2) released from cells. Under the optimal conditions (the applied potentials, the flow rate and the sample volume), the device exhibits high sensitivity toward H2O2. Linear relationships could be achieved between electrochemical responses and the concentration of H2O2 ranging from 1 nM to 1 mM. The excellent analytical performance of the SSWM-based device could be attributed to the pass-through mode based on the mesh microstructure and intrinsic catalytic properties for H2O2 by stainless steel. This approach could be further successfully extended for screening of H2O2 released from HeLa cells with electrochemical responses linear to the number of cells in a range of 3 - 1.35 × 104 cells with an injection volume of 30 µL. This study revealed the potential of mesh electrodes in electrochemical flow injection analysis for cellular function and pathology and its possible extension in cell counting and on-line analysis.

Rapid assays of SARS-CoV-2 virus and noble biosensors by nanomaterials.

The COVID-19 outbreak caused by SARS-CoV-2 in late 2019 has spread rapidly across the world to form a global epidemic of respiratory infectious diseases. Increased investigations on diagnostic tools are currently implemented to assist rapid identification of the virus because mass and rapid diagnosis might be the best way to prevent the outbreak of the virus. This critical review discusses the detection principles, fabrication techniques, and applications on the rapid detection of SARS-CoV-2 with three categories: rapid nuclear acid augmentation test, rapid immunoassay test and biosensors. Special efforts were put on enhancement of nanomaterials on biosensors for rapid, sensitive, and low-cost diagnostics of SARS-CoV-2 virus. Future developments are suggested regarding potential candidates in hospitals, clinics and laboratories for control and prevention of large-scale epidemic.

Identification of large yellow croakers (Larimichthys crocea) scavenger receptor genes: Involvement in immune response to Pseudomonas plecoglossicida infection and hypoxia-exposure experiments.

Scavenger receptors (SRs) are pattern recognition receptors involved in the innate immune defense against pathogen infection in fish. However, there has not been much research done on teleosts. In this study, 18 members of the SR gene family were found in large yellow croaker. The identification of the SR gene family showed that the protein length of SR members in large yellow croaker were quite different, and most SR genes were distributed in nuclear and endoplasmic. The evolutionary relationship, exon/intron structure and motif analysis revealed that members of the SR gene family were highly conserved. The results of the expression profiles after Pseudomonas plecoglossicida infection and hypoxia-exposure demonstrated that SR members were involved in inflammatory reactions. Especially, COLEC12 and SCARF1 exhibited substantial changes in response to both P. plecoglossicida and hypoxia stress, indicating their possible immunological functions. The result of this study revealed that SR genes played a vital part in the innate immune response of large yellow croaker, and would give important details for a deeper comprehension of the SR gene family's regulation mechanism under various conditions in large yellow croaker.

Vibration-enhanced disposable electroanalytical platform for selective analysis of tryptophan in fruits based on molecular imprinting.

Although electrochemical detection based on molecular imprinting polymers (MIP) could dramatically improve the selectivity, the procedure is time-consuming because of the essential incubation step. In addition, current MIP electrochemical detections were not suitable for analysis of microliter-level sample solutions, limiting their applications for real samples. This investigation aims at applying vibration to enhance efficiency of MIP electrochemical detection of 20 µL sample solutions. MIP analysis of Tryptophan (Trp) was used as the model with disposable MIP electrodes prepared by electrochemical polymerization of o-phenylenediamine on carbon ink coated on stainless steel sheets. The MIP electrode was integrated in a 3D-printed analytical device for vibration-enhanced electrochemical detection of Trp. Our results showed that this vibration-enhanced strategy could significantly increase electrochemical responses of Trp at the same incubation time. Such improvement might be attributed to the enhanced mass transfer at the surface of the working electrode brought by vibration. It needs to be emphasized that this strategy is suitable for analysis of sample solutions with the volume of microliters, which is superior to normal stirring in MIP electrochemical detection. Our approach could be successfully utilized for differentiation of Trp in different fruits, opening more opportunities for MIP electrochemical detection of real samples. The enhanced efficiency by vibration could pave foundation for extensive practical MIP detection of sample solutions at the level of microliters.

Synergistically Enhanced Photocatalytic Degradation by Coupling Slow-Photon Effect with Z-Scheme Charge Transfer in CdS QDs/IO-TiO2 Heterojunction.

Lower light absorption and faster carrier recombination are significant challenges in photocatalysis. This study introduces a novel approach to address these challenges by anchoring cadmium sulfide quantum dots (CdS QDs) on inverse opal (IO)-TiO2, which increases light absorption and promotes carriers' separation by coupling slow-photon effect with Z-scheme charge transfer. Specifically, the IO-TiO2 was created by etching a polystyrene opal template, which resulted in a periodic structure that enhances light absorption by reflecting light in the stop band. The size of CdS quantum dots (QDs) was regulated to achieve appropriate alignment of energy bands between CdS QDs and IO-TiO2, promoting carrier transfer through alterations in charge transfer modes and resulting in synergistic-amplified photocatalysis. Theoretical simulations and electrochemical investigations demonstrated the coexistence of slow-photon effects and Z-scheme transfer. The system's photodegradation performance was tested using rhodamine B as a model. This novel hierarchical structure of the Z-scheme heterojunction exhibits degradability 7.82 and 4.34 times greater than pristine CdS QDs and IO-TiO2, respectively. This study serves as a source of inspiration for enhancing the photocatalytic capabilities of IO-TiO2 and broadening its scope of potential applications.

Electron-Rich Triazine-Conjugated Microporous Polymers for the Removal of Dyes from Wastewater.

Conjugated microporous polymers (CMP) as porous functional materials have received considerable attention due to their unique structures and fascinating properties for the adsorption and degradation of dyes. Herein, a triazine-conjugated microporous polymer material with rich N-donors at the skeleton itself was successfully synthesized via the Sonogashira-Hagihara coupling by a one-pot reaction. These two polymers had Brunauer-Emmett-Teller (BET) surface areas of 322 and 435 m2g-1 for triazine-conjugated microporous polymers (T-CMP) and T-CMP-Me, respectively. Due to the porous effects and the rich N-donor at the framework, it displayed a higher removal efficiency and adsorption performance compared to cationic-type dyes and selectivity properties for (methylene blue) MB+ from a mixture solution of cationic-type dyes. Furthermore, the T-CMP-Me could quickly and drastically separate MB+ and (methyl orange) MO- from the mixed solution within a short time. Their intriguing absorption behaviors are supported by 13C NMR, UV-vis absorption spectroscopy, scanning electron microscopy, and X-ray powder diffraction studies. This work will not only improve the development of porous material varieties, but also demonstrate the adsorption or selectivity of porous materials for dyes from wastewater.

Vibration for enhancement of electrochemical analysis of biomolecules in a droplet on the rough surface of a disposable working electrode.

Although electrochemical detection of microliters-level solutions is attractive for analysis of low-amount biological samples, its performance could be weakened by limited mass transfer due to low Reynolds number and laminar flow. Herein we designed a 3D-printed electroanalytical device to apply vibration for improvement of mass transfer during electrochemical detection. In our approach, the droplet-size sample solution containing Indole-3-acetic acid (IAA, as a model) was directly applied on the effective surface of a disposable working electrode. We demonstrated that vibration could enhance electrochemical responses of IAA more on the rough surface than on the smooth surface of the working electrodes. After optimization, the sensitivity for electrochemical detection of a 20-µL droplet under vibration with the voltage of 7 V increased more than 100% compared with the static condition. The enhanced electrochemical responses brought by vibration could be achieved reproducibly, which could be ascribed to improved mass transfer. Our strategy could be practically applied for differentiation of IAA in different tissues of Marchantia polymorpha with enhanced responses. This study suggested that vibration might become a simple and effective method to improve mass transfer in analysis of microliter-volume solutions, which might be extended for more biochemical assays.

Acid etching followed by water soaking: a top-down strategy to induce highly reactive substrates for electrocatalysis.

A top-down strategy using acid etching followed by water soaking is utilized to in situ synthesize autologous NiFe LDH nanosheets on NiFe foam without other metal ions, oxidizing agents or heating steps. The NiFe foam serves as both the metal source and substrate, and the obtained nanosheets are firmly anchored on the foam. The obtained ultrathin nanosheet arrays could greatly increase the electrocatalytic active sites. This factor together with the synergistic effect between Fe and Ni simultaneously leads to an enhanced catalytic effect for water splitting and urea oxidation. This strategy could be scaled up to pave a viable way for low-cost fabrication of highly efficient electrodes for electrocatalysis.

Sandwich-Type Electrochemiluminescence Immunosensor Based on CDs@dSiO2 Nanoparticles as Nanoprobe and Co-Reactant.

In general, co-reactants are essential in highly efficient electrochemiluminescence (ECL) systems. Traditional co-reactants are usually toxic, so it is necessary to develop new environmentally friendly co-reactants. In this work, carbon dots (CDs) were assembled with dendritic silica nanospheres (CDs@dSiO2 NPs) to form a co-reactant of Ru(bpy)32+. Subsequently, a sandwich immunosensor for detecting human chorionic gonadotropin (HCG) was constructed based on CDs@dSiO2 NPs as co-reactants, the nanoprobe loaded with the secondary antibody, and Ru(bpy)32+ as a luminophore. In addition, compared to directly as a signal probe, the luminophore Ru (bpy)32+ as a part of the electrolyte solution is simpler in this work. The immunosensor has an extremely low limit of detection of 0.00019 mIU/mL. This work describes the synthesis of low-toxic, efficient, and environmentally friendly CDs, which have become ideal co-reactants of Ru(bpy)32+, and proposes an ECL immunosensor with excellent stability and selectivity, which has great potential in clinical applications.

Interplay of Dual-Proton Transfer Relay to Achieve Full-Color Panel Luminescence in Excited-State Intramolecular Proton Transfer (ESIPT) Fluorophores.

A new design was applied for the facile synthesis of pure organic photoluminescent molecules with dual excited-state intramolecular proton transfer (ESIPT) sites. In this novel class of emitters, full-color panel emission from blue, green, and yellow to red, including white light, can be achieved in different solvents as modulated by the enol-keto(1st)-keto(2nd) tautomer emissions. A comprehensive transient photophysical study verifies that keto(1st) and keto(2nd) have a precursor (<0.8 ps)-successor (∼20 ps)-relayed absorbance relationship, and then a fast equilibrium between the two is established, resulting in dual emissions in the nanosecond scale (∼1900 ps). Through the research on copper ions' selective PL response, the dual-ESIPT mechanism was further verified; in addition, the study of solid-state PL changes upon the stimulus of organic vapor manifests the potential application sensitivity of the molecules as dual-ESIPT sensors. Theoretical results including reaction potential energy surface analyses manifest the fact that dual-proton transfer goes along a sequential route with a smaller energy barrier, firmly supporting the experimental results. An intrinsic system that undergoes intramolecular double proton relayed transfer is thus established for the achievement of much broadened optical responses and full-color display, providing reference for the design and application of advanced dual-ESIPT optical materials.

A 3D-printed analytical device seamlessly integrating sample treatment for electrochemical detection of IAA in Marchantia polymorpha.

Because of the pivotal point of Marchantia polymorpha (M. polymorpha) in plant evolution, its auxin (mainly indole-3-acetic acid, IAA) levels could provide useful evidence for the study of the evolution of IAA. However, M. polymorpha could not be easily pretreated for electrochemical detection because they are at the entry level of land plants. Herein, we designed a three-dimensional (3D)-printed analytical device for seamless integration of sample treatment and electrochemical detection. Specifically, the electrochemical cell could be used as a mortar in which a tiny plant sample could be ground with a 3D-printed pestle, followed by mixing with the buffer solution under vibration for electrochemical detection of IAA with a disposable working electrode at the bottom of the cell. Using our strategy, the limits of quantification could reach 0.05 µmol L-1 after optimization of parameters. We were able to demonstrate that IAA in different tissues of wild-type and mutant M. polymorpha could be successfully differentiated after they were treated with the 3D-printed analytical device. The obtained results were comparable to the samples blended with zirconium beads while the differences of IAA levels in different tissues of M. polymorpha agreed well with previous reports. This study suggested the potential of sample treatment integrated with electrochemical detection for analysis of IAA using the 3D printing techniques and their possible applications in the research of plants and other fields.

Human Gingival Stem Cells Prevent Diabetes in NOD Mice by Reducing Follicular B Cells / 中山大学学报(医学科学版)

BackgroundType 1 diabetes is caused by a chronic immune response that destroys islet beta cells， resulting in elevated blood glucose. Mesenchymal stem cells can prevent and treat the development of diabetes and its complications. However， little is known about the effects and potential mechanisms of Gingival mesenchymal stem cells （GMSCs） in preventing diabetes. The aim of this study is to investigate the mechanism of GMSCs in preventing type 1 diabetes in mice and to find targets for clinical treatment of diabetes. MethodsWe injected human GMSCs into NOD mice to observe the trend of blood glucose， observed the survival of pancreatic β-cells by immunohistochemistry， and detected the change of immune cells in the spleen of mice by flow analysis. Finally， the immune cells in NOD mice were transfused into NOD-SCID mice to observe the onset of diabetes in NOD-SCID mice. ResultsGMSCs significantly reduced the incidence of diabetes in NOD mice， with 64% of control mice developing diabetes at 27 weeks of age compared with 35% in the GMSC group， P=0.013. The percentage of Follicular B cells（FO B cell） in the spleen of GMSCs-treated mice decreased from （52.2±4.1）% to （43.2±5.3）%， P=0.008， while other types of immune cells did not change significantly. The immunohistochemical results showed that GMSCs could effectively improve the survival of pancreatic β-cells， which could continuously produce insulin to control blood glucose. Finally， we found the spleen cells transfusion could prevent the development of diabetes in NOD-SCID mice. ConclusionGMSCs can reduce diabetes in mice by reducing FO B cells in the spleen.

Protein-protein interactions between RUNX3 and ZEB1 in chronic lung injury induced by methamphetamine abuse.

Methamphetamine (MA) is the most common and highly addictive substance abuse drug. Runt-related transcription factor 3 (RUNX3) and Zinc finger E-box-binding homeobox 1 (ZEB1) are associated with lung inflammation and fibrosis. However, the protein-protein interactions (PPIs) between RUNX3 and ZEB1 and its involvement in MA-induced chronic lung injury is still unclear. In this study, we evaluated lung injury using echocardiography, hematoxylin and eosin staining, and western blot analysis. The viability of alveolar epithelial cells (AECs) was assessed using cell counting kit-8. Molecular Operating Environment software, Search Tool for the Retrieval of Interacting Genes/Proteins database, co-immunoprecipitation, assay and confocal immunofluorescence assay were used to predict and identify the PPIs between RUNX3 and ZEB1. The expression of RUNX3 and ZEB1 were knockdown in AECs using siRNA. The results revealed that MA exposure increased the peak blood flow velocity of the pulmonary artery and the acceleration time of pulmonary artery blood flow. Further, exposure to MA also causes adhesion and fusion of the alveolar walls and altered AEC activity. A decrease in the expression of RUNX3 and an increase in the expression of ZEB1 and its downstream signaling molecules were observed on MA exposure. The PPIs between RUNX3 and ZEB1 were identified. Further, an increase in the protein binding rate of RUNX3-ZEB1 was observed in MA-induced lung injury. These results show interactions between RUNX3 and ZEB1. RUNX3 protects against lung injury; however, ZEB1 expression and the PPIs between ZEB1 and RUNX3 has deleterious effects on chronic lung injury induced by MA exposure. Our results provide a new therapeutic approach for the treatment of chronic lung injury due to MA exposure.

Surface Reconstruction of an FeNi Foam Substrate for Efficient Oxygen Evolution.

Designing earth-abundant electrocatalysts that are highly active, low-cost, and stable for the oxygen evolution reaction (OER) is crucial for electrochemical water splitting. However, in conventional electrode fabrication strategies, NiFe layered double hydroxide (NiFe LDH) catalysts are usually coated onto substrates as external components, which suffers from poor conductivity, easily detaches from the substrate, and hinders their long-term utilization. Herein, the surface-reconstruction strategy is used to synthesize in situ autologous NiFe LDH to increase the surficial active sites numbers. The FeNi foam (FNF) serves as both the metal source and substrate, and the obtained NiFe LDH nanosheets (NSs) are firmly anchored in the monolithic FNF. What needs to be emphasized is that the strategy does not involve any high-temperature or high-pressure processes, apart from a cost-effective etching and a specified drying treatment. The nanostructure of NiFe LDH and the synergistic effect between Fe and Ni simultaneously lead to an enhanced catalytic effect for the OER. Remarkably, the sr-FNF46 requires only an ultralow overpotential of 283 mV to achieve a current density of 100 mA cm-2 for the OER in 1 M KOH electrolyte, and exhibits excellent stability. Thus, the obtained electrode holds promise for electrocatalytic applications. Finally, the formation mechanism of NiFe LDH NSs due to surface reconstruction is investigated and discussed in detail.

Identification of circRNA-miRNA-mRNA networks to explore the molecular mechanism and immune regulation of postoperative neurocognitive disorder.

Postoperative neurocognitive disorder (PND) is a common complication in older patients. However, its pathogenesis has still remained elusive. Recent studies have shown that circular RNA (circRNA) plays an important role in the development of neurodegenerative diseases, such as PND after surgery. CircRNA, as a competitive endogenous RNA (ceRNA), mainly acts as a molecular sponge for miRNA to "adsorb" microRNA (miRNA) and to reduce the inhibitory effects of miRNAs on target mRNA. The sequencing data of circRNA were obtained from the Gene Expression Omnibus (GEO) database. By bioinformatic methods, circAtlas, miRDB, miRTarBase and miRwalk databases were applied to construct circRNA-miRNA-mRNA networks and screen differentially expressed mRNAs. To improve the accuracy of the data, we randomly divided aging mice into control (non-PND group) and PND groups, and used high-throughput sequencing to analyze their brain hippocampal tissue for analysis. Three key genes were cross-detected in the data of both groups, which were Unc13c, Tbx20 and St8sia2 (as hub genes), providing new targets for PND treatment. According to the results of the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, immune cell infiltration analysis, gene set enrichment analysis (GSEA), Connectivity Map (CMap) analysis, quantitative real-time polymerase chain reaction (qRT-PCR), the genes that were not related to the central nervous system were removed, and finally, mmu_circ_0000331/miR-1224-3p/Unc13c and mmu_circ_0000406/miR-24-3p/St8sia2 ceRNA networks were identified. In addition, the CMap method was used to select the top 4 active compounds with the largest negative correlation absolute values, including cimaterol, Rucaparib, FG-7142, and Hydrocortisone.

A protein with broad functions: damage-specific DNA-binding protein 2.

Damage-specific DNA-binding protein 2 (DDB2) was initially identified as a component of the damage-specific DNA-binding heterodimeric complex, which cooperates with other proteins to repair UV-induced DNA damage. DDB2 is involved in the occurrence and development of cancer by affecting nucleotide excision repair (NER), cell apoptosis, and premature senescence. DDB2 also affects the sensitivity of cancer cells to radiotherapy and chemotherapy. In addition, a recent study found that DDB2 is a pathogenic gene for hepatitis and encephalitis. In recent years, there have been few relevant literature reports on DDB2, so there is still room for further research about it. In this paper, the molecular mechanisms of different biological processes involving DDB2 are reviewed in detail to provide theoretical support for research on drugs that can target DDB2.

Disposable stainless steel working electrodes for sensitive and simultaneous detection of indole-3-acetic acid and salicylic acid in Arabidopsis thaliana leaves under biotic stresses.

The detection of phytohormones in real time has attracted increasing attention because of their critical roles in regulating the development and signaling of plants, especially in defense against biotic stresses. Herein, stainless steel sheet electrodes modified with carbon cement were coupled with paper-based analysis devices for direct and simultaneous detection of salicylic acid (SA) and indole-3-acetic acid (IAA) in plants. We demonstrated that the excellent conductivity of stainless steel sheet electrodes enabled us to simultaneously differentiate IAA and SA at a level of 10 nM. With our approach, the content of IAA and SA in Arabidopsis thaliana leaves infected or not infected with Pst DC3000 could be rapidly quantified at the same time. Our experimental results on differentiation of IAA and SA at different time points showed that there were antagonistic interactions between the IAA and SA after infection of Arabidopsis leaves with Pst DC3000. By offering a cost-effective approach for rapid and sensitive detection of IAA and SA, this study suggests that electrochemical detection can be used in the study and development of precision agriculture technology.

Stereoselective Synthesis of Tetrasubstituted Olefins via Visible-Light-Promoted Iodine-Mediated Homo-Coupling of Diazo.

A visible-light-promoted iodine-mediated homo-coupling of diazo was first described. A series of tetrasubstituted olefins were synthesized in high yields and with low to high Z-selectivities from phenyldiazoacetates. For 3-diazooxindoles, isoindigo derivatives were provided in moderate to high yields and with excellent E-selectivities. Experimental results showed that the reaction proceeded through a diiodo intermediate. The synthetic usefulness of this reaction was illustrated by the synthesis of maleimide derivatives and dispiro epoxy.

A sandwich-type electrochemical immunosensor for CYFRA 21-1 based on probe-confined in PtPd/polydopamine/hollow carbon spheres coupled with dendritic Au@Rh nanocrystals.

A signal-on sandwich-like electrochemical immunosensor was built for determination of cytokeratin 19 fragments 21-1 (CYFRA 21-1) in non-small cell lung cancer (NSCLC) by confining electroactive dye (e.g., methylene blue, MB) as a probe for amplifying signals. Specifically, core-shell gold@rhodium dendritic nanocrystals (Au@Rh DNCs) behaved as a substrate for primary antibody and accelerate interfacial electron transfer. Besides, hollow carbon spheres (HCSs) were subsequently modified with polydopamine (PDA) and PtPd nanoparticles for sequential integration of the secondary antibody and confinement of MB as a label, termed as MB/PtPd/PDA/HCSs for clarity. The built sensors showed a broad linear range (100 fg mL-1 ~ 100 ng mL-1) for detection of CYFRA 21-1 with an ultra-low detection limit (31.72 fg mL-1, S/N = 3), coupled with satisfactory performance in human serum samples. This work can be explored for assays of other proteins and provides some constructive insights for early and accurate diagnosis of NSCLC.

[Sensitivity Comparison Experiment of Four Testing Methods for Helicobacter pylori].

Objective: To measure with standard microbiology methods the sensitivity of 4 commonly used testing methods for Helicobacter pylori (Hp) and to conduct a comparative study of the correlations and differences across the 4 methods. Methods: With the Hp standard strain (SS1) as the reference, colony forming units (CFU) as the units of quantitative analysis for detection performance, and gradient dilution of SS1 suspension as the simulation sample, we measured the sensitivity of 4 Hp testing methods, including bacterial culture, rapid urease test, antigen test, and quantitative fluorescent PCR. CFU values at different concentrations corresponding to the 4 commonly used Hp testing methods were documented and the correlations and differences were analyzed accordingly. Results: The sensitivity of Hp bacterial culture, rapid urease test, antigen test and quantitative fluorescent PCR was 2.0×10 CFU/mL, 2.0×10 5 CFU/mL, 2.0×10 5 CFU/mL, and 2.0×10 2 CFU/mL, respectively. Conclusion: The testing turnover time and sensitivity of different laboratory methods for Hp testing varied significantly. The quantitative fluorescent PCR and bacterial culture both showed relatively high sensitivity, but bacterial culture has complicated operation procedures and is too time-consuming. The rapid urease test and antigen test both were simple and quick to perform, but showed low sensitivity. For clinical and laboratory testing of Hp, appropriate testing method that can identify the corresponding changes of Hp should be selected according to the actual testing purpose.