Stacking and ridge regression-based spectral ensemble preprocessing method and its application in near-infrared spectral analysis.

Spectral preprocessing techniques can, to a certain extent, eliminate irrelevant information, such as current noise and stray light from spectral data, thereby enhancing the performance of prediction models. However, current preprocessing techniques mostly attempt to find the best single preprocessing method or their combination, overlooking the complementary information among different preprocessing methods. These preprocessing techniques fail to maximize the utilization of useful information in spectral data and restrict the performance of prediction models. This study proposed a spectral ensemble preprocessing method based on the rapidly developing ensemble learning methods in recent years and the ridge regression (RR) model, named stacking preprocessing ridge regression (SPRR), to address the aforementioned issues. Different from conventional ensemble learning methods, the proposed SPRR method applied multiple different preprocessing techniques to the original spectral data, generating multiple preprocessed datasets. These datasets were then individually inputted into RR base models for training. Ultimately, RR still served as the meta-model, integrating the output results of each RR base model through stacking. This approach not only produced diversity in base models but also achieved higher accuracy and lower computational complexity by using a single type of base model. On the apple spectral dataset collected by our team, correlation analysis showed significant complementary information among the data produced by different preprocessing techniques. This provided robust theoretical support for the proposed SPRR method. By introducing the currently popular averaging ensemble preprocessing method in a comparative experiment, the results of applying the proposed SPRR method to six datasets (apple, meat, wheat, olive oil, tablet, and corn) demonstrated that compared to the single preprocessing method and averaging ensemble preprocessing method, SPRR yielded the best accuracy and reliability for all six datasets. Furthermore, under the same conditions of the training and test datasets, the proposed SPRR method demonstrated better performance than the four commonly used ensemble preprocessing methods.

Gut-Derived Trimethylamine N-Oxide Promotes CCR2-Mediated Macrophage Infiltration in acute kidney injury.

BACKGROUND: Inflammation is crucial in the development of AKI and subsequent CKD following renal ischemia-reperfusion (IR) injury. Gut microbiota metabolites trigger inflammation and affect IR-induced renal damage. Yet, the driving factors and mechanisms are unclear. Trimethylamine N-oxide (TMAO), a gut-derived choline metabolite, is a strong pro-inflammatory factor that increases in patients with AKI and CKD. We hypothesized that TMAO can promote renal injury caused by IR. METHODS: Mice subjected to unilateral renal IR to induce AKI and CKD were fed a high-choline diet to observe the effects of TMAO on kidney inflammation, fibrosis, and macrophage dynamics. RESULTS: A choline-rich diet altered the gut microbiota and elevated TMAO levels, which exacerbated IR-induced AKI and subsequent CKD. Single-cell analysis identified a distinct subset of CCR2+ macrophages derived from monocytes as key responders to TMAO, intensifying immune cell interactions and worsening renal injury. TMAO promoted sustained CCR2 expression after IR, increasing macrophage infiltration. CCR2 deletion and antagonist RS-102895 improved TMAO-induced inflammation and fibrosis, alleviated renal injury induced by IR. CONCLUSIONS: Our study provides valuable insights into the link between TMAO and IR-incited renal inflammation and fibrosis, emphasizing the critical role of TMAO-mediated macrophage infiltration via CCR2 as a key therapeutic target in the acute and chronic phase after IR.

A platinum glutamate acid complex as a peroxidase mimic: high activity, controllable chemical modification, and application in biosensors.

Recent strides in nanotechnology have given rise to nanozymes, nanomaterials designed to emulate enzymatic functions. Despite their promise, challenges such as batch-to-batch variability and limited atomic utilization persist. This study introduces Pt(Glu)2, a platinum glutamic acid complex, as a versatile small-molecule peroxidase mimic. Synthesized through a straightforward method, Pt(Glu)2 exhibits robust catalytic activity and stability. Steady-state kinetics reveal a lower Km value compared to that of natural enzymes, signifying strong substrate affinity. Pt(Glu)2 was explored for controllable chemical modification and integration into cascade reactions with natural enzymes, surpassing other nanomaterials. Its facile synthesis and seamless integration enhance cascade reactions beyond the capabilities of nanozymes. In biosensing applications, Pt(Glu)2 enabled simultaneous detection of cholesterol and alkaline phosphatase in human serum with high selectivity and sensitivity. These findings illustrate the potential of small molecule mimetics in catalysis and biosensing, paving the way for their broader applications.

RNA-Seq Analysis of ceRNA-Related Networks in the Regulatory Metabolic Pathway of Mice with Diabetic Nephropathy Subjected to Empagliflozin Intervention.

OBJECTIVE: We conducted bioinformatics analysis of the gene chip data of empagliflozin for diabetic nephropathy (DN). The differentially expressed genes (DEGs) between DN and control mice and between DN and DN treated with empagliflozin (DNE) mice were screened to explore the related metabolic pathogenesis and predict the potential competing endogenous RNA (ceRNA)-related networks' metabolic mechanism of the empagliflozin effect on DN. METHODS: The intersection of DEGs in mice between the control and DN groups and between the DN and DNE groups was selected. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses were performed, and the metabolic items involving the most genes in the coregulation were considered. A protein-interaction network was constructed with the STRING website. Cytoscape software and its plug-ins were utilised to analyse the hotspot differential genes. The noncoding RNAs in which the differential genes may play a role were obtained from the miRanda, miRDB, and TargetScan databases to establish network diagrams. RESULTS: Analysis of the diabetes and control groups showed that 424 genes were upregulated and 354 were downregulated. In the analysis of DEGs between the DN and diabetic groups, the comparison between the diabetic and empagliflozin groups showed that 430 genes were upregulated and 84 were downregulated. The co-downregulated enrichment results were primarily reflected in various metabolic disorders, including glucose metabolism, lipid metabolism, amino acid metabolism, and others. The co-upregulated genes were associated with the inflammatory response, apoptosis, and cell senescence. This finding indicated that empagliflozin may inhibit the progression of diabetic nephropathy by inhibiting inflammation, apoptosis, and senescence. The key genes and related mechanisms of noncoding RNA were determined through Cytoscape analysis and the prediction of common DEGs in metabolic items. CONCLUSIONS: The analysis of DEGs and key core genes in this study enhanced our understanding of the effect of empagliflozin on the pathogenesis of DN and provided more potential gene targets and application ideas for DN treatment.

A dual-mode strategy based on ß-galactosidase and target-induced DNA polymerase protection for transcription factor detection using colorimetry and a glucose meter.

In this work, we report a novel dual-mode method for the highly specific and sensitive detection of transcription factors (TFs) via the integration of Klenow polymerase protection induced by target-specific recognition, cascade-signal amplification using the hybridization chain reaction (HCR) and CRISPR/Cas12a system, and dual-signal transduction mediated by ß-galactosidase (ß-gal) and two substrates. A dual-mode signal-sensing interface was constructed by immobilizing the oligo DNA probe (P1) tethered ß-gal in a 96-well plate. A hairpin H1 with the ability to initiate HCRs was designed to contain the TF binding site. The binding between the TF and H1 protected the H1 from being extended by the Klenow fragment. After thermal denaturation, the reserved H1 launched the HCR and the HCR products activated CRISPR/Cas12a to cleave P1 and reduce the ß-gal on the sensing interface, and thus the contents of the TFs and the corresponding signals mediated by the catalysis of ß-gal showed a correlation. This work was the first attempt at utilizing ß-gal for dual-signal transduction. It is a pioneering study to utilize the HCR-CRISPR/Cas12a system for dual-mode TF sensors. It revealed that DNA polymerase protection through the binding of TF and DNA could be applied as a new pattern to develop TF sensors.

RNA-Seq Analysis of ceRNA-Related Networks in the Regulatory Metabolic Pathway of Mice with Diabetic Nephropathy Subjected to Empagliflozin Intervention

Objective: We conducted bioinformatics analysis of the gene chip data of empagliflozin for diabetic nephropathy (DN). The differentially expressed genes (DEGs) between DN and control mice and between DN and DN treated with empagliflozin (DNE) mice were screened to explore the related metabolic pathogenesis and predict the potential competing endogenous RNA (ceRNA)-related networks’ metabolic mechanism of the empagliflozin effect on DN. Methods: The intersection of DEGs in mice between the control and DN groups and between the DN and DNE groups was selected. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses were performed, and the metabolic items involving the most genes in the coregulation were considered. A protein-interaction network was constructed with the STRING website. Cytoscape software and its plug-ins were utilised to analyse the hotspot differential genes. The noncoding RNAs in which the differential genes may play a role were obtained from the miRanda, miRDB, and TargetScan databases to establish network diagrams. Results: Analysis of the diabetes and control groups showed that 424 genes were upregulated and 354 were downregulated. In the analysis of DEGs between the DN and diabetic groups, the comparison between the diabetic and empagliflozin groups showed that 430 genes were upregulated and 84 were downregulated. The co-downregulated enrichment results were primarily reflected in various metabolic disorders, including glucose metabolism, lipid metabolism, amino acid metabolism, and others. The co-upregulated genes were associated with the inflammatory response, apoptosis, and cell senescence. This finding indicated that empagliflozin may inhibit the progression of diabetic nephropathy by inhibiting inflammation, apoptosis, and senescence. The key genes and related mechanisms of noncoding RNA were determined through Cytoscape analysis and the prediction of common DEGs in metabolic items (AU)

Multifunctional Gold-Silver-Carbon Quantum Dots Nano-Hybrid Composite: Advancing Antibacterial Wound Healing and Cell Proliferation.

The urgent need for innovative materials that effectively eliminate bacteria while promoting cell growth to accelerate wound healing has led to the exploration of new options, as current antimicrobial nanoparticles often exhibit high cytotoxicity, which hinders wound closure. In this study, a nano-hybrid composite, named gold-silver-carbon quantum dots (AuAg-CDs), was prepared by embedding gold and silver nanoclusters into carbon dots. The AuAg-CDs nano-hybrid composite demonstrates remarkable biocompatibility, displays potent antibacterial activity, and possesses a unique capability to promote cell proliferation. By physically disrupting bacterial membranes and promoting mammalian cell proliferation, this composite emerges as a highly promising material for wound healing applications. The underlying mechanism of the multifunctional AuAg-CDs was investigated through comprehensive analyses encompassing cell morphology, bacterial membrane potential, levels of reactive oxygen species (ROS), and adenosine triphosphate (ATP) production in both bacterial and mammalian cells. Additionally, AuAg-CDs were incorporated into alginate to create a hydrogel wound dressing, which underwent evaluation using animal models. The results underscore the remarkable potential of the AuAg-CDs wound dressing in facilitating the proliferation of wound fibroblasts and combating bacterial infections. The significance of designing multifunctional nanomaterials to address the challenges associated with pathogenic bacterial infections and regenerative medicine is highlighted by this study, paving the way for future advancements in these fields.

EDTA-functionalized silica nanoparticles as a conditioning agent for dentin bonding using etch-and-rinse technique.

OBJECTIVE: This study investigated the possibility of using ethylenediaminetetraacetic acid functionalized silica nanoparticles (EDTA-SiO2) as a dentin-conditioning agent using etch-and-rinse technique to promote the durability of dentin bonding. METHODS: The SiO2-EDTA were synthesized by N- [(3- trimethoxysilyl) propyl] ethylenediamine triacetic acid (EDTA-TMS) and SiO2 (50 nm), then characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM). The capacity of SiO2-EDTA to chelate calcium ions from dentin was examined by inductively coupled plasma-optic emission spectrometry (ICP-OES). The dentin surfaces conditioned with SiO2-EDTA were detected by field emission scanning electron microscopy (SEM), TEM and microhardness testing. For dentin bonding, dentin surfaces were adopted wet- or dry-bonding technique and bonded with adhesive (AdperTM Single Bond2) and applied composite resin (Filtek Z350) on them. The durability of dentin bonding was evaluated by mircotensile bond strength test, in-situ zymography and nanoleakage testing. RESULTS: FTIR, TGA and XPS results showed that SiO2-EDTA contained N element and carboxyl groups. SEM, TEM and microhardness results indicated that SiO2-EDTA group created extrafibrillar demineralization and retained more intrafibrillar minerals within dentin surface. In the dentin bonding experiment, SiO2-EDTA group achieved acceptable bond strength, and reduced the activity of matrix metalloproteinase and nanoleakage along bonding interface. CONCLUSION: It was possible to generate a feasible dentin conditioning agent (SiO2-EDTA), which could create dentin extrafibrillar demineralization and improve dentin bond durability. CLINICAL SIGNIFICANCE: This study introduces a new dentin conditioning scheme based on SiO2-EDTA to create extrafibrillar demineralization for dentin bonding. This strategy has the potential to be used in clinic to promote the life of restoration bonding.

Dual-mode photoelectrochemical/electrochemical sensor based on Z-scheme AgBr/AgI-Ag-CNTs and aptamer structure switch for the determination of kanamycin.

A "signal-on" dual-mode aptasensor based on photoelectrochemical (PEC) and electrochemical (EC) signals was established for kanamycin (Kana) assay by using a novel Z-scheme AgBr/AgI-Ag-CNTs composite as sensing platform, an aptamer structure switch, and K3[Fe(CN)6] as photoelectron acceptor and electrochemical signal indicator. The aptamer structure switch was designed to obtain a "signal-off" state, which included an extended Kana aptamer (APT), one immobilized probe (P1), and one blocking probe (P2) covalently linked with graphdiyne oxide (GDYO) nanosheets. P1, P2, and aptamer formed the double helix structure, which resulted in the inhibited photocurrent intensity because of the weak conductivity of double helix layer and serious electrostatic repulsion of GDYO towards K3[Fe(CN)6]. In the presence of Kana, APT specifically bound to the target and dissociated from P1 and P2, and thus, a "signal-on" state was initiated by releasing P2-GDYO from the platform. Based on the sensing platform and the aptamer structure switch, the dual-mode aptasensor realized the linear determination ranges of 1.0 pM-2.0 µM with a detection limit (LOD) of 0.4 pM (for PEC method) and 10 pM-5.0 µM with a LOD of 5 pM (for EC method). The aptasensor displayed good application potential for Kana test in real samples.

Anti-phospholipase A2 receptor antibody levels at diagnosis predicts outcome of TAC-based treatment for idiopathic membranous nephropathy patients.

BACKGROUND: Idiopathic membranous nephropathy (iMN) is recognized as an organ-specific autoimmune disease, mainly caused by anti-PLA2R antibody. This study aimed to study between anti-PLA2R antibody level at diagnosis and the response to tacrolimus (TAC)-based treatment in iMN patients. METHODS: This was a retrospective cohort study including 94 kidney biopsy-proven MN patients with positive anti-PLA2R antibody at diagnosis from May 2017 to September 2021 in our center. All iMN patients received the TAC regimen as the initial immunosuppressive therapy. All patients were divided into two groups according to anti-PLA2R antibody titer at diagnosis: high-level group (> 150 RU/ml; n = 42) and low-level group (≤ 150 RU/ml; n = 52). The association between anti-PLA2R antibody levels and clinical outcomes was assessed using the Kaplan-Meier method. RESULTS: The low density lipoprotein in the high-level group was significantly higher than low-level group at diagnosis, otherwise, serum albumin was significantly lower than low-level group; however, there was no significant difference in creatinine levels between two groups. The remission rates were significantly higher in the low-level group than high-level group after treatment with TAC for 12, 18, or 24 months (all P < 0.05). After 12 months of treatment with TAC, 82.7% of the patients in the low-level group achieved complete remission (CR) or partial remission (PR) (mean, 6.52 ± 0.53 months). However, 38.1% of the patients in high-level group achieved CR or PR (mean, 9.86 ± 0.51 months). Moreover, CR rate at 12 months in the high-level group was only 4.7% (mean, 11.88 ± 0.63 months). The infection frequency in the high-level group (35.6%) was higher than the low-level group (20%) during the TAC treatment, although there was no significant difference (P = 0.065). There were 19% patients who had end-stage kidney disease (ESKD), and 7.1% of patients died of ESKD in the high-level group during the follow-up period. CONCLUSION: Anti-PLA2R antibody level above 150 RU/ml at diagnosis can predict a poor treatment response and outcome of TAC treatment in iMN patients, who may not benefit from TAC or other calcineurin inhibitor regimens as the initial treatment.

Changes in the spectrum of kidney diseases: a survey of 2803 patients from 2010 to 2018 at a single center in southeastern China.

Primary glomerular disease was the leading cause of chronic kidney disease (CKD) in China; however, changes in the economy and environment introduce variations in the spectrum of kidney diseases. This study aimed to analyze renal biopsy data to inform disease prevention and public health interventions. In this retrospective cohort study, data from 2,803 consecutive renal biopsies conducted at our center between January 2010 and December 2018 were analyzed. The sample was disaggregated by age and the date of biopsy to facilitate analysis. Primary glomerulonephritis (PGN) is the most frequent (81.84%) finding, followed by secondary glomerulonephritis (SGN; 15.38%), tubulointerstitial nephritis (15.38%), and others (1.57%). IgA nephropathy (IgAN), idiopathic membranous nephropathy (iMN), and minimal change disease were the primary causes of PGN. Among PGN cases, the incidence of iMN arose, especially among those aged ≥ 60 years old, during the observation period. Contrary to the case of iMN, the proportion of IgAN in PGN trended downward, continuously, and at length. Moreover, IgAN mainly affected those aged 25-44 years old and less so those aged ≥ 60 years old. Lupus nephritis, Henoch-Schönlein purpura nephritis, and diabetic nephropathy (DN) were key causes of SGN. A ratio reversal between infectious disease and chronic disease dramatically changed SGN patterns. In the past year, the incidence of hepatitis B-related nephritis has constantly declined; however, the proportion of DN among SGN had steadily increased. The incidence of iMN significantly increased during these years. Among SGN cases, the proportion of DN has increased.

Sensitive fluorescence detection of pathogens based on target nucleic acid sequence-triggered transcription.

Accurate identification of mutant pathogens derived from genetic polymorphisms is highly desired in clinical diagnosis. However, current detection methods based on Watson-Crick hybridization suffers from false positives due to the cross-reactivity of wild-type sequences. In this study, we developed an accurate identification of mutant pathogens by combining programmable DNAzyme and target nucleic acid sequence-triggered transcription. Single nucleotide variants (SNVs) are the most plentiful type of mutations in the genome. High specificity to discriminate SNV was first achieved by rational design of dual-hairpin DNA structure and DNAzyme's capability of site-specific cleavage. T7 RNA polymerase-mediated transcription amplification was introduced to exponentially increase the sensitivity by encompassing T7 promoter sequence into the dual-hairpin DNA structure. The design of this biosensor is fast and straightforward without many computational steps, and the highly sensitive biosensor can detect not only SNVs but also occasional insertions and large deletions in the genome. We showed that the assay could rapidly detect COVID-19 variant and methicillin-resistant Staphylococcus aureus (MRSA), and the limit of detection is 0.96 copy/µL. The modular design of functional DNA enables this biosensor be easily reconfigured and is useful diagnosis of emerging infectious diseases caused by mutant pathogens.

A novel missense mutation in complement factor I predisposes patients to atypical hemolytic uremic syndrome: a case report.

BACKGROUND: Atypical hemolytic uremic syndrome, also called the nondiarrheal form of hemolytic uremic syndrome, is a rare disease characterized by the triad of thrombocytopenia, Coomb's test-negative microangiopathic hemolytic anemia, and acute renal failure. Approximately 60% of cases of atypical hemolytic uremic syndrome are associated with deficiencies of the complement regulatory protein, including mutations in complement factor H, complement factor I, or the membrane co-factor protein. CASE PRESENTATION: We report the case of a 26-year-old Asian man who presented with pulmonary infection, elevated blood pressure, microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. Renal biopsy revealed diffuse capillary fibrin deposition, endothelial swelling, and arteriole narrowing like "onion skinning" consistent with thrombotic microangiopathy. Bidirectional sequencing of CFH, CFHR5, CFHR1, CFI, DGKE, CFB, and MCP confirmed that the patient was heterozygous for a novel missense mutation, p.Cys67Phe, in CFI. This patient had rapid evolution to end-stage renal disease and needed renal replacement therapy. Plasma exchange seemed inefficacious in this patient. CONCLUSIONS: This report confirms the importance of screening patients with atypical hemolytic uremic syndrome for mutations in genes involved in complement system to clarify the diagnosis and demonstrates the challenges in the management of these patients.

Tunable graphdiyne for DNA surface adsorption: affinities, displacement, and applications for fluorescence sensing.

Graphdiyne (GDY) adsorbed DNA probes have been used as a fluorescent sensing platform, but topics including DNA adsorption affinities, DNA probe displacement, and fluorescence quenching ability were rarely researched. Herein, the adsorption affinity of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) on a tremella-like GDY was tuned by modulating the surface chemistry of GDY. The fluorescence quenching ability of GDY with different oxidation degrees was compared. The nonspecific displacement of DNA probes on GDY was studied. Under the same concentrations, GDY with low oxidation degree exhibited stronger adsorption affinity and higher adsorption capacity to both ssDNA and dsDNA than highly oxidized GDY. DNA adsorbed on low-oxidized GDY was more resistant to displacement by other DNAs. Protein showed strong interaction with different GDY and could displace DNA probes on GDY. Based on these findings, an ideal GDY with proper oxidation degree, exhibiting high surface affinity for ssDNA and low affinity for dsDNA, was used as scavenger of redundant ssDNA fluorescent probe in an enzyme-assisted amplification system for sensitive ochratoxin (OTA) detection. This study has enhanced our fundamental understanding of DNA adsorption by GDY. It also provided a rational way to apply GDY for fluorescence sensing in a complicated system.

Visual Quantitative Detection of Glutathione and Cholesterol in Human Blood Based on the Thiol-Ene Click Reaction-Triggered Wettability Change of the Interface.

Herein, we proposed an innovative visual quantitative sensing strategy based on thiol-ene click chemistry and the capillary action principle. A triethoxyvinylsilane (VTEO)- or mercaptopropylsilatrane (MPS)-modified interface was prepared for analyte recognition. Target analyte molecules containing thiol groups or C═C double bonds are coupled to the VTEO- or MPS-modified inner surface of the glass capillary tube via a thiol-ene click reaction, respectively. Then, the molecular recognition events were transformed into the wettability change of the inner wall of the glass capillary. The concentration of the target molecules was quantified by reading the height change of the water column in the capillary tube. As a proof of concept, this strategy was successfully used to build visual quantitative sensors for detecting glutathione and cholesterol. In addition, this strategy showed a good anti-interference ability to complex biological fluids and realized sensitive glutathione (GSH) and cholesterol detection in real human blood samples.

A cathode photoelectrochemical assay of terminal deoxynucleotidyl transferase activity based on Ag-AgI-CNTs composite and surface multisite strand displacement amplification.

Photocathode-based assay is anti-interference for real sample detection. Photocathode produces low photocurrent signal and gives rise to poor sensitivity. Herein, a novel cathode photoelectrochemical (CPEC) sensing platform based on Ag-AgI-CNTs as photocathode material and K3[Fe(CN)6] as photoelectron acceptor was established. Since [Fe(CN)6]3- effectively accepted photoelectrons from Ag-AgI-CNTs, it greatly enhanced the CPEC response. Combining a surface multisite strand displacement amplification (SMSDA) strategy, the CPEC platform was applied for the activity assay of terminal deoxynucleotidyl transferase (TdT). In this proposal, oligo dT primer tethered on CPEC platform was in-situ extended to generate a polyA tail. Then the polyA tail formed a stable multi-point hybrid structure with the adjacent oligo dT. After launching the SMSDA, the CPEC platform was covered by more elongated polynucleotide chains and network, which acutely hampered the photoelectron transfer (eT) between photocathode and electron acceptor and caused a reduced photocurrent. The CPEC sensor possessed a satisfactory linear response from 6 × 10-5-0.1 U and a low detection limit of 1.1 × 10-5 U. The strategy offered a more specific and sensitive method for TdT activity assay. It was feasible in the field of TdT-based biochemical research, drug screening, and disease diagnosis.

De Novo Development of a Universal Biosensing Platform by Rapid Direct Native Protein Modification.

An innovative biosensing assay was developed for simplified, cost-effective, and sensitive detection. By rapid, direct treatment of target proteins with iron porphyrin (TPPFe) in situ, a carboxyl group of amino acid conjugates with an Fe atom of the TPPFe molecule, forming a stable protein complex. We have shown that this complex not only maintains the integrity and functions of original proteins but also acquires peroxidase activity that can turn TMB to a comparably visible signal like that in ELISA. This study is unique since such conversion is difficult to achieve with standard chemical modification or molecular biology methods. In addition, the proposed immunoassay is superior to traditional ELISA as it eliminates an expensive and complicated cross-linking process of an enzyme-labeled antibody. From a practical point of view, we extended this assay to rapid detection of clinically relevant proteins and glucose in blood samples. The results show that this simple immunoassay provides clinical diagnosis, food safety, and environmental monitoring in an easy-to-implement manner.

Monitoring biothiols dynamics in living cells by ratiometric fluorescent gold carbon dots.

Changes in biothiols, including glutathione (GSH) and cysteine, are closely related to a variety of diseases and cellular functions. Real-time monitoring intracellular GSH and cysteine dynamics in living cells are important for understanding pathophysiological processes. In this study, a stable heterostructure of dual-emission fluorescent gold carbon dots (GCDs) consisting of carbon skeleton and gold nanoclusters was prepared, which emits strong blue fluorescence and weak yellow-orange fluorescence. The blue fluorescence of the as-prepared GCDs has no response to biothiols including GSH and cysteine, but a turn-on yellow-orange fluorescence would be appeared accompanied by the biothiols reacting with the GCDs. Ratiometric fluorescent bioimaging therefore may be established based on the unique GCDs to detect GSH and cysteine levels in living cells. Moreover, real time monitoring GSH and cysteine levels in various cell lines in living cells at different growth stages was realized. The concentration of GSH in cancer cells is higher than that of normal cells, however, the level of cysteine in normal cells is consistently higher than in cancer cells at different stages of cell growth. The fluorescent GCDs probe provides a promising tool for tracking on regulation of GSH and cysteine dynamics by physiological environments due to the capability for real-time quantitation of GSH and cysteine.

Real-Time Imaging Redox Status in Biothiols and Ferric Metabolism of Cancer Cells in Ferroptosis Based on Switched Fluorescence Response of Gold Carbon Dots.

Ferroptosis is an iron-dependent form of regulated cell death. In this study, a ratiometric fluorescent probe, gold carbon dots (GCDs) consisting of carbon skeleton and gold nanoclusters, was used for in situ imaging to monitor redox status in biothiols (glutathione and cysteine) and ferric metabolism of cancer cells in ferroptosis. The as-prepared GCDs can selectively respond to biothiols, interestingly, the fluorescence may be switched to sense ferric ions without interference by biothiols under proper conditions. The robust GCDs-probe exhibits excellent photobleaching resistance and can reversibly respond to intracellular biothiols/ferric ion with high temporal resolution. The 8 h real-time imaging of living cells was employed to track the fluctuation of biothiols, showing the change of redox status in ferroptosis. In addition, release of ferric ions in cells was monitored. The real-time imaging of depletion of biothiols and release of ferric ion in cells indicates the GCDs-probe can monitor how the ferroptosis regulates redox status in biothiols and ferric metabolism.