One-pot hydrothermal synthesis of silicon, nitrogen co-doped carbon dots for enhancing enzyme activity of acid phosphatase (ACP) to dopamine and for cell imaging.

Developing water-soluble nanomaterials with high photoluminescence emission and high yield for biological analysis and imaging is urgently needed. Herein, water-soluble blue emitting silicon and nitrogen co-doped carbon dots (abbreviated as Si-CDs) of a high photoluminescence quantum yield of 80 % were effectively prepared with high yield rate (59.1 %) via one-step hydrothermal treatment of N-[3-(trimethoxysilyl)propyl]ethylenediamine (DAMO) and trans-aconitic acid. Furthermore, the Si-CDs demonstrate environmental robustness, photo-stability and biocompatibility. Given the importance of the potentially abnormal levels of acid phosphatase (ACP) in cancer diagnosis, developing a reliable and sensitive ACP measurement method is of significance for clinical research. The Si-CDs unexpectedly promote the catalytic oxidation of ACP on dopamine (DA) to polydopamine under acidic conditions through the produced reactive oxygen species (ROS). Correspondingly, a fluorescence response strategy using Si-CDs as the dual functions of probes and promoting enzyme activity of ACP on catalyzing DA was constructed to sensitively determine ACP. The quantitative analysis of ACP displayed a linear range of 0.1-60 U/L with a detection limit of 0.056 U/L. The accurate detection of ACP was successfully achieved in human serum through recovery tests. As a satisfactory fluorescent probe, Si-CDs were successfully applied to fluorescent imaging of A549 cells in cytoplasmic with long-term and safe staining. The Si-CDs have the dual properties of outstanding fluorescent probes and auxiliary oxidase activity, indicating their great potential in multifunctional applications.

A facile fluorescence method for the effective detection of ampicillin using antioxidant carbon dots with specific fluorescent response to ˙OH.

Monitoring methods for beta-lactam (ß-lactam) antibiotics, especially for ampicillin (AMP), with simple operation and sensitivity for realtime applications are highly required. To address this need, antioxidant carbon dots (E-CDs) with excellent fluorescence properties were synthesized using citric acid and ethylenediamine as raw materials. With a quantum yield of 81.97%, E-CDs exhibited a specific and sensitive response to ˙OH. The quenched fluorescence of E-CDs by the formed ˙OH could be restored through a competition reaction with AMP. Leveraging the signal-quenching strategy of E-CDs, H2O2, and Fe2+, a fluorescence signal-on strategy was developed using AMP as the fluorescence recovery agent for the sensitive detection of AMP. The mechanism of the quenching of E-CDs by ˙OH was attributed to the damaging effect of ˙OH on E-CDs. Under optimal conditions, the detection limit of this method for AMP was determined to be 0.38 µg mL-1. This method was successful in drug quality control and the spiked detection of AMP in lake water, milk, and sea cucumber, presenting a viable option for convenient and rapid antibiotic monitoring methods.

Characterization and evaluation of cytotoxic and antimicrobial activities of cyclotides from Viola japonica.

Cyclotides are a type of defense peptide most commonly found in the Violaceae family of plants, exhibiting various biological activities. In this study, we focused on the Viola japonica as our research subject and conducted transcriptome sequencing and analysis using high-throughput transcriptomics techniques. During this process, we identified 61 cyclotides, among which 25 were previously documented, while the remaining 36 were designated as vija 1 to vija 36. Mass spectrometry detection showed that 21 putative cyclotides were found in the extract of V. japonica. Through isolation, purification and tandem mass spectrometry, we characterized and investigated the activities of five cyclotides. Our results demonstrated inhibitory effects of these cyclotides on the growth of Acinetobacter baumannii and Bacillus subtilis, with minimum inhibitory concentrations (MICs) of 4.2 µM and 2.1 µM, respectively. Furthermore, time killing kinetic assays revealed that cyclotides at concentration of 4 MICs achieved completely bactericidal effects within 2 h. Additionally, fluorescence staining experiments confirmed that cyclotides disrupt microbial membranes. Moreover, cytotoxicity studies showed that cyclotides possess cytotoxic effects, with IC50 values ranging from 0.1 to 3.5 µM. In summary, the discovery of new cyclotide sequences enhances our understanding of peptide diversity and the exploration of their activity lays the foundation for a deeper investigation into the mechanisms of action of cyclotides.

Insights into the Roles of Natural Killer Cells in Osteoarthritis.

Osteoarthritis (OA) is now widely acknowledged as a low-grade inflammatory condition, in which the intrinsic immune system plays a significant role in its pathogenesis. While the involvement of macrophages and T cells in the development of OA has been extensively reviewed, recent research has provided mounting evidence supporting the crucial contribution of NK cells in both the initiation and advancement of OA. Accumulated evidence has emerged in recent years indicating that NK cells play a critical role in OA development and progression. This review will outline the ongoing understanding of the utility of NK cells in the etiology of OA, focusing on how NK cells interact with chondrocytes, synoviocytes, osteoclasts, and other immune cells to influence the course of OA disease.

Effectively synthesized functional Si-doped carbon dots with the applications in tyrosinase detection and lysosomal imaging.

There has been significant interest in the preparation and versatile applications of carbon dots (CDs) due to their immense potential value in sensors and imaging. In this work, silicon-doped green carbon dots (Si-CDs) with high quantum yield and rich epoxypropyl were effectively synthesized. Given the clinical diagnostic importance of abnormal levels of tyrosinase (TYR), sensitive detection of TYR is significant for clinical research. A fluorescence signal-off strategy with Si-CDs as probe was constructed to determine TYR based on the oxidation of dopamine by TYR. The detection ranges of this method were 0.01-1.5 and 10-30 U/mL with the detection limit of 0.0046 U/mL, the lower limit of quantification (LLOQ) was 0.01 U/mL, and TYR was successfully and accurately monitored in human serum. Additionally, due to the role of lysosomes in cellular regulatory processes, including TYR levels and fluorescence stability characteristics of Si-CDs in acidic conditions, it was envisaged to use Si-CDs as probe to establish real-time monitoring of lysosomes. According to fluorescence colocation analysis, Si-CDs had intrinsic lysosomal targeting ability to HepG2 and L-02 (with Pearson correlation coefficients were 0.90 and 0.91, respectively). The targeting of Si-CDs to lysosomes was due to the acidophilic effect of the epoxypropyl on its surface.

A Fluorescence Biosensor for Tyrosinase Activity Analysis Based on Silicon-Doped Carbon Quantum Dots.

Tyrosinase (TYR) plays a pivotal role in the biosynthesis of melanin, and its activity level holds critical implications for vitiligo, melanoma cancer, and food nutritional value. The sensitive determination of TYR activity is of great significance for both fundamental research and clinical investigations. In this work, we successfully synthesized silicon-doped carbon quantum dots (Si-CQDs) through a one-pot hydrothermal method with trans-aconitic acid as carbon source and N-[3-(trimethoxysilyl)propyl]ethylenediamine as the dopant, exhibiting remarkable fluorescence quantum yield (QY) and photostability. Correspondingly, Si-CQDs were used as a probe to construct a sensitive, rapid, and user-friendly fluorescence method for TYR detection. The method relied on the oxidation of isoprenaline (ISO) by TYR, where Si-CQDs were employed as a highly efficient probe. The testing mechanism was the internal filtering effect (IFE) observed between Si-CQDs and the oxidative system of ISO and TYR. Under the optimized conditions, the fluorescence strategy exhibited a detection range of 0.05-2.0 U/mL for TYR with a limit of detection (LOD) of 0.041 U/mL. Furthermore, we successfully demonstrated the accurate determination of TYR levels in human serum, showcasing the promising potential of this method in various practical scenarios.

Detection of Doxycycline Using Carbon Quantum dots as Probe Based on Internal Filtering Effect.

The establishment of a convenient and effective detection method for doxycycline (DC) holds significant importance in drug monitoring and drug residue assessment. In this work, carbon quantum dots (CQDs) with excellent and stable luminescence performance (the quantum yield of CQDs was 21.8%) were synthesized by the melting method and employed as probes to monitor the fluorescence intensity variations before and after the introduction of DC. A fluorescence analytical method based on the internal filtration effect (IFE) was developed for DC determination. The mechanism of DC quenching CQDs was verified using fluorescence lifetime tests, absorption spectroscopy, and evaluation of internal filtration parameters. After optimizing experimental conditions, it was found that the DC concentration (CDC) exhibited a good linear relationship with the fluorescence quenching efficiency ((F0-F)/F0) of CQDs in the range of 5-30 µM. The fitted linear equation was Y = 0.01249*CDC+0.03625, R2 = 0.9987, and the detection limit was 2.343 µM (n = 8). This developed method has been successfully applied to accurately determine DC concentrations in both doxycycline hydrochloride tablets and human serum samples. It stands out for its simplicity, rapidity, and acceptable detection performance. Due to its advantages, this method holds great promise for application in the biomedical field for monitoring DC drug concentrations and ensuring quality control.

Decorated Polyetheretherketone Implants with Antibacterial and Antioxidative Effects through Layer-by-Layer Nanoarchitectonics Facilitate Diabetic Bone Integration with Infection.

Patients suffering diabetic bone defects still need some new and effective strategies to achieve enhanced prognostic effects. Although medical implants are the common treatment of bone defects, the excessive oxidative stress and high risk of bacterial infection in diabetes mellitus lead to a higher risk of implant failure. To improve the healing ability of diabetic bone defects, herein, polyetheretherketone (PEEK) was modified through a developed layer-by-layer (LBL) construction strategy to obtain multifunctional PEEK (SP@(TA-GS/PF)*3) by the assembly of tannic acid (TA), gentamicin sulfate (GS) and Pluronic F127 (PF127) on the basis of prepared porous PEEK through sulfonation (SPEEK). The prepared SP@(TA-GS/PF)*3 exhibited sustained antimicrobial activity and enhanced the differentiation of osteoblast (MC3T3-E1) for needed osteogenesis. Moreover, SP@(TA-GS/PF)*3 scavenged excessive oxidative stress to promote the growth of H2O2 damaged HUVEC with enhanced secretion of VEGF for neovascularization. In addition, the remarkable in vivo outcomes of angiogenesis and osseointegration were revealed by the subcutaneous implant model and bone tissue implant model in diabetic rats, respectively. The in vitro and in vivo results demonstrated that modified PEEK with multifunction can be an attractive tool for enhancing bone integration under diabetic conditions, underpinning the clinical application potential of modified implants for diabetic osseointegration.

A signal-off fluorescent strategy for deferasirox effective detection using carbon dots as probe and Cu2+ as medium.

The content of deferasirox (DEF) in plasma is significant in ß-thalassemia patient that needs long-term transfusion therapy, while the effective and simple strategy for DEF monitoring is still limited. The carbon dots (CDs) prepared from citric acid monohydrate and glutathione exhibit weakly modulated fluorescence intensity to several common metal ions containing Cu2+. Interestingly, the process of interaction of Cu2+ and DEF forms the chelation of Cu2+ and DEF (Cu-DEF) with the absorbance wavelength of DEF at 320 nm shifting to 332 nm for Cu-DEF. And the obtained Cu-DEF will effectively quench CDs through inner filter effect (IFE). Accordingly, a facile signal-off fluorescent method based on CDs as probe is developed for DEF detection using Cu2+ as medium. And the proposed method exhibits linear range of 0.5-20 µg/mL with the detection limit of 0.33 µg/mL for DEF under the optimized conditions. Moreover, the developed assay is further expanded to test the content of DEF in dispersible tablet and plasma with accuracy and reproducibility. Such cost-effective and sensitive fluorescent assay just through simple mixing operation present a valuable strategy for drug monitoring.

Antibacterial activity of positively charged carbon quantum dots without detectable resistance for wound healing with mixed bacteria infection.

Widespread bacterial infection and the spread of antibiotic resistance exhibit increasing threat to the public and thus require new antibacterial strategies. Carbon quantum dots (CQDs) have been extensively investigated to play fluorescent, catalytic roles and even potential biomedical functions containing sterilization. However, synthetic understanding of the interaction of CQDs and bacteria, the exhibition of antibacterial ability, and the risk of resistance evolution remain lacking. Herein, a simple one-pot method was fabricated to prepare positively charged CQDs (PC-CQDs) as a broad-spectrum antibacterial agent. PC-CQDs possessed effective antibacterial activity against all tested Gram-positive, Gram-negative, and drug-resistant bacteria. Investigation of the antibacterial mechanism of PC-CQDs indicated that small-sized PC-CQDs functionalized with -NH2 and -NH induced strong adherence behavior on the bacterial cell membrane. Moreover, the entry of PC-CQDs caused conformational changes in the genes and generation of reactive oxygen species in the bacteria. Safety evaluation illustrated that PC-CQDs did not trigger detectable drug resistance or hemolysis. Furthermore, PC-CQDs effectively promoted the antibacterial treatment of mixed Staphylococcus aureus and Escherichia coli infected wound in rats with low in vivo toxicity. These results suggested that PC-CQDs are a potential antibacterial candidate for real wound healing applications in complex bacterial infections and even resistant bacteria-caused infections.

Integration of fluorescent polydopamine nanoparticles on protamine for simple and sensitive trypsin assay.

As an important protease, trypsin (TRY) has been identified as a key indicator of various diseases. A simple and sensitive strategy for TRY detection by using an environment-friendly biosafe probe is significant. Herein, we introduced negatively charged fluorescent polydopamine nanoparticles (PDNPs) with 4.8 nm diameter obtained through a controllable method as an effective probe for TRY. PDNPs exhibited excellent fluorescence property but integrated with protamine (Pro) to form an aggregation-caused quenching system via a static quenching mechanism. The quenching mechanism of Pro to PDNPs revealed the significant effect of the surface charge, functional groups, and appropriate size of PDNPs on quenching process. Given the specific hydrolysis of Pro by TRY, PDNPs were released from the quenching integration of PDNPs and Pro (PDNPs-Pro) and recovered their fluorescence. Thus, a fluorescence sensor for TRY with a linear range of 0.01 and 0.1 µg/mL and a detection limit of 6.7 ng/mL was developed without the disturbing from other proteases. Compared with other TRY assays, the biosensor based on PDNPs-Pro has the advantages of simple operation, environmental friendliness, and high sensitivity. This specific controlled-synthesis PDNPs would open up a new window for the extended application of fluorescent nanomaterials in biomedicine based on fluorescence changes induced by biological interaction.

Ratiometric fluorescence assay based on carbon dots and Cu2+-catalyzed oxidation of O-phenylenediamine for the effective detection of deferasirox.

The monitoring of deferasirox (DEF) has important clinical roles in patients who need iron excretion. However, analytical methods with practicability and simplicity are limited. Moreover, ratiometric fluorescence strategies based on Förster resonance energy transfer (FRET) from carbon dots (CDs) as a donor are rarely reported as a drug monitor. In this work, CDs with an appropriate emitting wavelength at 480 nm and excitation around 370 nm were prepared by hydrothermal approach and HCl post-treatment. O-Phenylenediamine (OPD) can be oxidized by Cu2+ to produce yellow fluorescent 2,3-diaminophenazine (oxOPD) in the system of Cu2+ and OPD (Cu-OPD). Correspondingly, a remarkable FRET from CDs to oxOPD in the system of CDs, Cu2+ and OPD (CDs-Cu-OPD) was fabricated with the quenching illustration of CDs, but emitting property of oxOPD. Attributed to the chelation ability of DEF on Cu2+, the inhibitory effects of DEF on the Cu2+-triggered oxidative capability reduced the FRET system by the decreased oxOPD. Thus, the recovered CDs at F 480 and decreased oxOPD at F 560 were found through a ratiometric mode by the addition of DEF in CDs-Cu-OPD for the DEF assay. The FRET behavior of CDs and oxOPD in CDs-Cu-OPD was proved clearly through the calculation of the association constant, binding constant, number of binding sites, and the distance between the donor and acceptor. Furthermore, this ratiometric method exhibited promising analytical performance for DEF with the application in real samples. The implementation of this work expands the application field of CDs and OPD oxidation in drug monitoring, and even other biological analyses through ratiometric strategy.

Higher Blood Vascular Cell Adhesion Molecule-1 is Related to the Increased Risk of Cardiovascular Events in Chronic Obstructive Pulmonary Disease.

Background: Vascular cell adhesion molecule-1 (VCAM-1) is associated with vascular-related inflammation and atherosclerosis. This study aimed to evaluate whether VCAM-1 can be used for an indication of increased risk of CV events in patients with COPD. Methods: Serum VCAM-1 levels were measured in 163 COPD patients. All COPD patients were prospectively followed up for a median period of 48 months (range=3-54). Cox proportional hazard analysis was performed to evaluate the prognostic value of serum VCAM-1 for predicting CV events. Results: Serum VCAM-1 levels were higher in COPD patients with CV events than in those without CV events (1174.4±365.3 ng/mL vs 947.8±293.2 ng/mL; P<0.001). The logistic regression analysis revealed that serum VCAM-1 (OR=1.750; 95% CI, 1.324-2.428; Ptrend=0.0012) was independently associated with CVD (cardiovascular disease) history after adjusting for age, sex, BMI, current smoker, current drinker, admission systolic and diastolic BP, LVEF and laboratory measurements in patients with COPD at baseline. The Kaplan-Meier analysis demonstrated that the rate of CV events was higher in COPD patients with serum VCAM-1 levels above the median (517.3 ng/mL) than in those with VCAM-1 levels below the median. The Cox proportional hazard analysis revealed that serum VCAM-1 (HR=2.617; 95% CI, 1.673-5.328; Ptrend<0.001) may be an independent prognostic factor for CV events in the COPD patients. Conclusion: Our results suggested that serum VCAM-1 was significantly and independently associated with CV events in COPD patients. The inflammatory marker may help clinicians predict CV complications early.

Depression and Perceived Stress, but Not Anxiety, are Associated with Elevated Inflammation in an Obese Adult Population.

BACKGROUND: Anxiety, depression and perceived stress are risk factors for adverse health problems. Inflammation participates in the development of chronic diseases such as psychiatric disorders. This study explored the relationships between inflammatory biomarkers and depression, anxiety and perceived stress in an obese adult population. METHODS: The relationships between psychological scores and inflammatory markers were analyzed. RESULTS: A higher BMI was not correlated with a higher anxiety score (P=0.152); however, BMI was positively associated with a higher depression score (P<0.001) and a higher perceived stress score (P<0.001). Multivariate linear regression analysis revealed that in participants with BMI≥30 and 25≤BMI<30, depression and perceived stress were significantly and independently associated with ICAM-1, E-selectin and CRP, but these associations were not observed in participants with BMI<25. The anxiety score was not associated with any inflammatory marker in any group of subjects, as determined by multivariate analysis. CONCLUSION: Depression and perceived stress were strongly associated with increased serum levels of pro-inflammatory markers, including ICAM-1, E-selectin and CRP, among a general obese population from the United States. These results further suggest that depression and perceived stress might also be chronic systemic inflammatory diseases.

Determination of chondroitin sulfate in synovial fluid and drug by ratiometric fluorescence strategy based on carbon dots quenched FAM-labeled ssDNA.

Chondroitin sulfate (CS) plays an increasingly important role in clinical settings and pharmacy quality control. However, sensitive and simple methods for CS detection remain limited. In this work, positively charged nitrogen doped carbon dots (P-NCDs) with internal luminescence and quenching property to FAM-labeled random-sequence ssDNA (F-ssDNA) were prepared by a simple heating method. P-NCDs attached and quenched F-ssDNA through electrostatic interaction to form the system of P-NCDs and F-ssDNA (P-NCDs/F-ssDNA) with retained fluorescence intensity of P-NCDs. The highly negatively charged CS reacted electrostatically with P-NCDs and then replaced F-ssDNA in P-NCDs/F-ssDNA to recover the fluorescence intensity of the original quenched F-ssDNA while retaining the internal fluorescence intensity of P-NCDs. Thus, by using restored F-ssDNA as the signal controlled by adding CS to P-NCDs/F-ssDNA, a ratiometric fluorescence strategy based on the retained fluorescence of P-NCDs as reference signal was fabricated through synchronous fluorescence spectrometry for the sensitive detection of CS. Under the optimal experimental conditions, a linear equation for CS was obtained for CS concentration within the range of 0.05-2.0 µg/mL. The method was also successfully applied for the accurate determination of CS in joint fluid samples of arthritic patients, chondroitin sulfate tablets, and chondroitin sulfate eye drops, suggesting its appreciable application potential in the clinic.

Facile Fluorescence Dopamine Detection Strategy Based on Acid Phosphatase (ACP) Enzymatic Oxidation Dopamine to Polydopamine.

Facile and effective detection of dopamine (DA) plays a significant role in current clinical applications. Substantially, special optical nanomaterials are important for fabricating easy-to-control, cheap, selective, and portable fluorescence DA sensors with superior performance. Herein, carbon dots (CDs) prepared from melting method were applied as signal to establish a simple but effective fluorescence strategy for DA determination based on the enzymatic activity of acid phosphatase (ACP), which induces DA to form polydopamine (pDA). The formed pDA caused by the enzymatic oxidization of ACP toward DA can interact with CDs through the inner filter effect. Such behavior effectively quenched the CDs' fluorescence. The degree of fluorescence quenching of CDs was positively correlated with the DA content. Under the optimized reaction conditions, the proposed fluorescence method exhibited a comparable analytical performance with other DA sensors with good selectivity. Furthermore, this method has been successfully applied to detect DA in DA hydrochloride injection and human serum samples. It shows that this method features potential practical application value and is expected to be used in clinical research.

Ratiometric fluorescence sensor based on carbon dots as internal reference signal and T7 exonuclease-assisted signal amplification strategy for microRNA-21 detection.

The expression level of miRNA-21 is closely related to the occurrence and development of cancer, especially in gastrointestinal cancer. Monitoring miRNA-21 has clinical application in the diagnosis and evaluation of gastrointestinal cancer. A turn-on ratiometric fluorescence bioassay based on the T7 exonuclease-mediated cyclic enzymatic amplification method was developed for miRNA-21 determination by using carbon dots (CDs) and FAM-labeled ssDNA as the signal source. CDs demonstrated the triple functions of built-in internal fluorescence, probe carrier, and quencher in this strategy. In the absence of miRNA-21, FAM-labeled ssDNA would be adsorbed and quenched by CDs. The addition of miRNA-21 induced cycle hydrolysis from the 5' end by the T7 exonuclease and then released the short-cleaved FAM-labeled oligonucleotides. Then, the increased FAM signal (FFAM) and the stable CD signal (FCDs) would be tested through a ratiometric routine for the quantification of miRNA-21. The FFAM/FCDs value showed a good linear relationship with the concentration of miRNA-21 in the range of 0.05-10 nM, and the detection limit for miRNA-21 was 1 pM with excellent selectivity and reproducibility. Furthermore, this sensor successfully evaluated the expression level of miRNA-21 in clinical blood samples from healthy individuals and gastrointestinal cancer patients, and the results were highly consistent with those of qRT-PCR, suggesting the great clinical application value in the diagnosis of cancer associated with miRNA-21 expression levels.

A simple fluorescence assay for trypsin through a protamine-induced carbon quantum dot-quenching aggregation platform.

The development of a simple detection strategy for trypsin (Try) is urgent, and is ascribed to the diagnostic value of Try in several diseases. Herein, a facile but effective fluorescence strategy for Try was developed based on the protamine (Pro)-induced aggregation of carbon quantum dots (CQDs). The fluorescence of negatively charged CQDs was quenched with Pro due to the assembly of CQDs and Pro (CQDs/Pro) through electrostatic interaction. However, the highly positively charged Pro, which is rich in basic arginine residues, was preferred to be hydrolyzed by Try. Try can induce the deaggregation of CQDs/Pro, thereby enabling the release of CQDs to restore the fluorescence intensity. Thus, the use of CQDs/Pro as a testing platform will be employed as a "turn-on" method for Try. In addition, the fluorescence-resuming response was proportional to Try, ranging from 25 ng mL-1 to 500 ng mL-1 with a limit of detection (LOD) of 8.08 ng mL-1. This "turn-on" fluorescence assay for Try was label-free, convenient, and relatively free of interference from coexisting substances. Actual applications for Try monitoring and trypsin inhibitor screening also illustrated the considerable prospect of CQDs in the clinical field, combined with the superiority of the simple mixing operation.