CRISPR-Hg: Rapid and visual detection of Hg2+ based on PCR coupled with CRISPR/Cas12a.

Mercury (Hg) is a notorious toxic heavy metal, causing neurotoxicity and liver damage, posing grave threats to human health and environmental safety. There is an urgent imperative for developing novel Hg2+ detection methods. In this work, we developed a CRISPR-based method for Hg2+ detection named CRISPR-Hg. A CRISPR/Cas12a system was employed and could be activated by the PCR product, generating fluorescence signals based on the trans-cleavage activity. CRISPR-Hg exhibited remarkable selectivity and specificity, achieving a detection limit of 10 pM and minimal interference with background signals. This approach has been successfully applied to detect Hg2+ in real samples, including water, soil, and mushroom. Ulteriorly, a portable device was devised to streamline the readout of fluorescence signals by a smartphone within 30 min. We offer an affordable, highly selective and visually interpretable method for Hg2+ detection, with the potential for broad application in Hg2+ monitoring for food safety and public health.

A xenogeneic extracellular matrix-based 3D printing scaffold modified by ceria nanoparticles for craniomaxillofacial hard tissue regeneration via osteo-immunomodulation.

Hard tissue engineering scaffolds especially 3D printed scaffolds were considered an excellent strategy for craniomaxillofacial hard tissue regeneration, involving crania and facial bones and teeth. Porcine treated dentin matrix (pTDM) as xenogeneic extracellular matrix has the potential to promote the stem cell differentiation and mineralization as it contains plenty of bioactive factors similar with human-derived dentin tissue. However, its application might be impeded by the foreign body response induced by the damage-associated molecular patterns of pTDM, which would cause strong inflammation and hinder the regeneration. Ceria nanoparticles (CNPs) show a great promise at protecting tissue from oxidative stress and influence the macrophages polarization. Using 3D-bioprinting technology, we fabricated a xenogeneic hard tissue scaffold based on pTDM xenogeneic TDM-polycaprolactone (xTDM/PCL) and we modified the scaffolds by CNPs (xTDM/PCL/CNPs). Through series ofin vitroverification, we found xTDM/PCL/CNPs scaffolds held promise at up-regulating the expression of osteogenesis and odontogenesis related genes including collagen type 1, Runt-related transcription factor 2 (RUNX2), bone morphogenetic protein-2, osteoprotegerin, alkaline phosphatase (ALP) and DMP1 and inducing macrophages to polarize to M2 phenotype. Regeneration of bone tissues was further evaluated in rats by conducting the models of mandibular and skull bone defects. Thein vivoevaluation showed that xTDM/PCL/CNPs scaffolds could promote the bone tissue regeneration by up-regulating the expression of osteogenic genes involving ALP, RUNX2 and bone sialoprotein 2 and macrophage polarization into M2. Regeneration of teeth evaluated on beagles demonstrated that xTDM/PCL/CNPs scaffolds expedited the calcification inside the scaffolds and helped form periodontal ligament-like tissues surrounding the scaffolds.

Construction of dental pulp decellularized matrix by cyclic lavation combined with mechanical stirring and its proteomic analysis.

The decellularized matrix has a great potential for tissue remodeling and regeneration; however, decellularization could induce host immune rejection due to incomplete cell removal or detergent residues, thereby posing significant challenges for its clinical application. Therefore, the selection of an appropriate detergent concentration, further optimization of tissue decellularization technique, increased of biosafety in decellularized tissues, and reduction of tissue damage during the decellularization procedures are pivotal issues that need to be investigated. In this study, we tested several conditions and determined that 0.1% Sodium dodecyl sulfate and three decellularization cycles were the optimal conditions for decellularization of pulp tissue. Decellularization efficiency was calculated and the preparation protocol for dental pulp decellularization matrix (DPDM) was further optimized. To characterize the optimized DPDM, the microstructure, odontogenesis-related protein and fiber content were evaluated. Our results showed that the properties of optimized DPDM were superior to those of the non-optimized matrix. We also performed the 4D-Label-free quantitative proteomic analysis of DPDM and demonstrated the preservation of proteins from the natural pulp. This study provides a optimized protocol for the potential application of DPDM in pulp regeneration.

Targeting and Microenvironment-Activated Nanoreactor for Diabetic Chronic Wound Healing via Multienzyme Cascade Reactions.

The development of cell-like nanoreactors with the ability to initiate biocatalytic cascades under special conditions holds tremendous potential for therapeutic applications. Herein, conformationally gated nanoreactors that respond to the acidic microenvironment of infected diabetic wounds were developed by cucur[8]bituril (CB[8])-based supramolecular assembly. The bioinspired nanoreactors exhibit not only self-regulated permeability and selectivity to control internal enzyme activities by substance exchange but also distinct binding specificities toward Gram-positive and Gram-negative bacteria via noncovalent modification with different ligands. The encapsulation of glucose oxidase (GOx), Fe3O4 nanozyme, and l-arginine (l-Arg) into the nanocarriers enables intelligent activation of multienzyme cascade reactions upon glucose (Glu) uptake to produce gluconic acid (GA) and hydrogen peroxide (H2O2), which is further converted into highly toxic hydroxyl radicals (·OH) for selective antibacterial activity. Moreover, acidic H2O2 promotes the oxidization of l-Arg, leading to the release of nitric oxide (NO). Consequently, this nanoreactor provides a multifunctional and synergistic platform for diabetic chronic wound healing by combining enzyme dynamic therapy with NO gas therapy to combat bacterial infections and inflammation under high blood Glu levels.

Personalized 3D-Printed Scaffolds with Multiple Bioactivities for Bioroot Regeneration.

Recent advances in 3D printing offer a prospective avenue for producing transplantable human tissues with complex geometries; however, the appropriate 3D-printed scaffolds possessing the biological compatibility for tooth regeneration remain unidentified. This study proposes a personalized scaffold of multiple bioactivities, including induction of stem cell proliferation and differentiation, biomimetic mineralization, and angiogenesis. A brand-new bioink system comprising a biocompatible and biodegradable polymer is developed and reinforced with extracellular matrix generated from dentin tissue (treated dentin matrix, TDM). Adding TDM optimizes physical properties including microstructure, hydrophilicity, and mechanical strength of the scaffolds. Proteomics analysis reveals that the released proteins of the 3D-printed TDM scaffolds relate to multiple biological processes and interact closely with each other. Additionally, 3D-printed TDM scaffolds establish a favorable microenvironment for cell attachment, proliferation, and differentiation in vitro. The 3D-printed TDM scaffolds are proangiogenic and facilitate whole-thickness vascularization of the graft in a subcutaneous model. Notably, the personalized TDM scaffold combined with dental follicle cells mimics the anatomy and physiology of the native tooth root three months after in situ transplantation in beagles. The remarkable in vitro and in vivo outcomes suggest that the 3D-printed TDM scaffolds have multiple bioactivities and immense clinical potential for tooth-loss therapy.

Phase estimation of a Mach-Zehnder interferometer via the Laguerre excitation squeezed state.

Quantum metrology has an important role in the fields of quantum optics and quantum information processing. Here we introduce a kind of non-Gaussian state, Laguerre excitation squeezed state as inputs of traditional Mach-Zehnder interferometer to examine phase estimation in realistic case. We consider the effects of both internal and external losses on phase estimation by using quantum Fisher information and parity detection. It is shown that the external loss presents a bigger effect than the internal one. The phase sensitivity and the quantum Fisher information can be improved by increasing the photon number and even surpass the ideal phase sensitivity by two-mode squeezed vacuum in a certain region of phase shift for realistic case. Our results can find significant practical applications in quantum metrology.

3D-bioprinted Recombination Structure of Hertwig's Epithelial Root Sheath Cells and Dental Papilla Cells for Alveolar Bone Regeneration.

Three-dimensional (3D) bioprinting is an emerging method for tissue regeneration. However, promoting the epithelial-mesenchymal interaction (EMI), while maintaining the characteristics of epithelial cells has always been a challenge in tissue engineering. Since EMI acts as a critical factor in bone regeneration, this study aims to promote EMI by recombining epithelial and mesenchymal cells through 3D bioprinting. Hertwig's epithelial root sheath (HERS) is a transient structure appeared in the process of tooth root formation. Its epithelial characteristics are easy to attenuate under appropriate culture environment. We recombined HERS cells and dental papilla cells (DPCs) through 3D bioprinting to simulate the micro-environment of cell-cell interaction in vivo. HERS cells and DPCs were mixed with gelatin methacrylate (GelMA) separately to prepare bio-inks for bioprinting. The cells/GelMA constructs were transplanted into the alveolar socket of Sprague-Dawley rats and then observed for 8 weeks. Hematoxylin and eosin staining, Masson staining, and immunohistochemical analysis showed that dimensional cultural pattern provided ideal environment for HERS cells and DPCs to generate mineralization texture and promote alveolar bone regeneration through their interactions. 3D bioprinting technology provides a new way for the co-culture of HERS cells and DPCs and this study is inspiring for future research on EMI model.

Multifunctional probe based on modified Ag&Mn:ZnInS QDs for dual-mode fluorescence and magnetic resonance imaging of intracellular glutathione.

Glutathione (GSH) plays vital roles in a variety of biological processes, and the development of simple and effective GSH detection method is an important research topic. Herein, a multifunctional probe based on Ag&Mn:ZnInS quantum dots (QDs) was developed for bimodal imaging of GSH. MnO2, as an efficient fluorescence quencher, was in-situ grown on the surface of QDs, and then modified with hyaluronic acid (HA) to improve the stability and targeted recognition capability of the probe due to the binding between HA and CD44 receptors. After MnO2 was deconstructed by GSH, the fluorescence of the probe was recovered and the generated Mn2+ could serve as good magnetic resonance imaging (MRI) contrast agent. Moreover, the near-infrared emission probe was successfully employed in living cell and zebrafish imaging due to its low toxicity and high anti-biological interference performance. This strategy provides a simple dual-mode fluorescence/MRI imaging of GSH, which may have a broad application in biological detection.

Mediating Effects of Adolescent Physical Activity, Self-Rated Health and Family Income.

Objective: To investigate and analyse the situation and relationship between adolescent physical activity and self-assessment of health to provide a reference for adolescent physical activity research. Methods: A questionnaire was used to investigate the physical activity and self-rated health of 1,804 adolescents aged 14-18 years. Results: There was a significant relationship between adolescents' physical activity and self-rated health. The coefficient was 0.109 (P < 0.01) in urban areas and 0.127 (P < 0.01) in rural areas. At the same time, it was found that when family income was used as the intermediary variable between physical activity and self-rated health, the intermediary effect was 0.12 (P < 0.01), and the intermediary effect accounted for 25.97%. Conclusion: Adolescent obesity, physical activity, smoking, wellbeing and physical activity can affect adolescents' self-rated health status. At the same time, it is also found that adolescents' family income is an intermediary variable between physical activity and self-rated health. Suggestions: (1) Increase the methods of sports venues, sports organizations and sports activities, improve the possibility of teenagers participating in physical activities, and improve teenagers' self-rated health; (2) There is a large gap between the physical activity and self-rated health of urban and rural adolescents. Increasing the guidance of physical activity of adolescents in rural areas promotes the balance of self-rated health of urban and rural adolescents. (3) Family income is the intermediary variable of teenagers' physical activities and self-rated health. Reducing family expenditure through financial transfer payments or reducing taxes and fees can increase the level of teenagers' physical and mental health.

Ratiometric fluorescence and colorimetric dual-mode sensing platform based on carbon dots for detecting copper(II) ions and D-penicillamine.

A sensing platform with both ratiometric fluorescence and colorimetric responses towards copper(II) ions (Cu2+) and D-penicillamine (D-pen) was constructed based on carbon dots (CDs). o-Phenylenediamine (OPD) was employed as a chromogenic development reagent for reaction with Cu2+ to generate the oxidation product 2,3-diaminophenazine (oxOPD), which not only emits green fluorescence at 555 nm, but also quenches the blue fluorescence of CDs at 443 nm via the inner filter effect (IFE) and Förster resonance energy transfer (FRET). Additionally, oxOPD exhibits obvious absorption at 420 nm. Since the intense chelation affinity of D-pen to Cu2+ greatly inhibits the oxidation of OPD, the intensity ratio of fluorescence at 443 nm to that at 555 nm (F443/F555) and the absorbance at 420 nm (A420) were conveniently employed as spectral response signals to represent the amount of D-pen introduced into the testing system. This dual-signal sensing platform exhibits excellent selectivity and sensitivity towards both Cu2+ and D-pen, with low detection limits of 0.019 µM and 0.092 µM, respectively. In addition, the low cytotoxicity of the testing reagents involved in the proposed sensing platform facilitates its application for live cell imaging.

Sensitive ratiometric fluorescence probe based on chitosan carbon dots and calcein for Alkaline phosphatase detection and bioimaging in cancer cells.

Alkaline phosphatase (ALP) is a commonly used marker in clinical practice, and this enzyme is a key indicator for diagnosing various diseases. In this study, we describe the development of a reliable and novel fluorescent assay for ALP detection based on chitosan carbon dots (C-CDs, peak emission, 412 nm) and calcein (peak emission, 512 nm). In the presence of Eu3+ (which binds calcein), the fluorescence intensity of calcein is quenched. Utilizing the ALP-triggered generation of phosphate ions (PO43-) from the substrate p-nitrophenyl phosphate (pNPP), the Eu3+ ions bind PO43- (which shows a higher affinity toward Eu3+ than calcein), and the fluorescence of calcein is recovered. As a consequence, C-CDs fluorescence is decreased by inner filter effect (IFE). Exploiting these changes in the fluorescence intensity ratio of C-CDs and calcein, we developed a high sensitivity, accurate, and easily synthesized ratiometric fluorescence probe. Our novel fluorescent bioassay demonstrates good linear relationship in the 0.09-0.8 mU mL-1 range, with a low detection limit of 0.013 mU mL-1. The excellent applicability of this novel assay in HepG2 cells and human serum samples demonstrates that our novel method has excellent biomedical research and disease diagnosis prospects.

Xenoextracellular matrix-rosiglitazone complex-mediated immune evasion promotes xenogenic bioengineered root regeneration by altering M1/M2 macrophage polarization.

Xenogenic extracellular matrix (xECM)-based organ transplantation will be a promising approach to address the problem of donor shortage for allotransplantation, which has achieved great success in organ regeneration. However, current approaches to utilize xECM-based organ have limited capacity to yield the host a biofriendly microenvironment for long-term immunity homeostasis, compromising the application of these xenografts for repairing and replacing damaged tissues. As the key innate immune cells, macrophages directly determine the prognosis of xenografts in long term. However, it has not been fully elucidated that how to modulate their biological behavior for microenvironment homeostasis in tissue reconstruction. In this study, we report a robust strategy to impart an immunosuppressive surface to naturally sponge-like porous xECM scaffolds by loading rosiglitazone (RSG) to activate peroxisome proliferators receptors-γ (PPAR-γ). The resultant xECM-RSG complex, enabling RSG to be delivered sequentially and continuously to cells without obvious systemic side effects, is recognized as "self" to escape immune monitoring in local immunoregulation by downregulating the expression of proinflammatory NOS2+ M1 macrophages and oxygen species (ROS) through suppressing NF-κB expression, greatly facilitating the regeneration of enthesis anchoring between the transplanted xenograft and host in both heterotopic and orthotopic models. The newly formed bio-root is morphologically and biomechanically equivalent to native tooth root with a significant expression of odontogenic differentiation-related critical proteins. Therefore, the PPAR-γ-NF-κB axis activated by the xECM-RSG complex enables the xenografts to converse towards M2 macrophages with a modest immunosuppressive capacity for facilitating in xECM-based tissue or organ regeneration.

Novel Bradykinin Receptor Inhibitors Inhibit Proliferation and Promote the Apoptosis of Hepatocellular Carcinoma Cells by Inhibiting the ERK Pathway.

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. Studies have shown that bradykinin (BK) is highly expressed in liver cancer. We designed the novel BK receptor inhibitors J051-71 and J051-105, which reduced the viability of liver cancer cells and inhibited the formation of cancer cell colonies. J051-71 and J051-105 reduced cell proliferation and induced apoptosis in HepG2 and BEL-7402 cells, which may be due to the inhibition of the extracellular regulated protein kinase (ERK) signaling pathway. In addition, these BK receptor inhibitors reversed the cell proliferation induced by BK in HepG2 and BEL-7402 cells by downregulating B1 receptor expression. Inhibiting B1 receptor expression decreased the protein levels of p-ERK and reduced the malignant progression of HCC, providing a potential target for HCC therapy.

A neoteric dual-signal colorimetric fluorescent probe for detecting endogenous/exogenous hydrogen peroxide in cells and monitoring drug-induced hepatotoxicity.

Hydrogen peroxide (H2O2), one of the most important reactive oxygen species (ROS), can be generated endogenously in the liver and has been deemed as a biomarker for evaluating drug-induced liver injury (DILI). Therefore, it is highly crucial to construct an effective method for detecting H2O2 in the liver in order to evaluate DILI. Herein, a neoteric dual-signal colorimetric fluorescent probe XH-2 for sensing hydrogen peroxide was engineered and synthesized. Borate was grafted as a specific recognition group onto the fluorophore XH-1 (ΦF = 0.34) to establish a structurally unprecedented probe. The experimental results manifested that probe XH-2 (ΦF = 0.15) was able to detect hydrogen peroxide using a fluorescence method with an excellent linear range of 0-140 µM (R2 = 0.9974) and an especially low detection limit of 91 nM (λex/em = 570 nm/638 nm). In addition, the probe was capable of monitoring hydrogen peroxide in a colorimetric manner with the linear range of 0-110 µM (R2 = 0.9965). Furthermore, the specificity, applicability in serum (98.6-109.1%) and indirect detection of glucose make the probe XH-2 a superior probe. Based on its low cytotoxicity, the probe was successfully applied to monitor endogenous/exogenous hydrogen peroxide and quantitatively determine the concentration level of hydrogen peroxide at a range of 0-120 µM (R2 = 0.9859) in HepG2 cells. Ultimately, the probe could effectively monitor the level of hydrogen peroxide during DILI in HepG2 cells.

A novel water-soluble near-infrared fluorescent probe for monitoring mitochondrial viscosity.

Mitochondria, the main source of energy of cells, play a significant role in aerobic respiration process. Some stimulants can result in changes of mitochondrial microenvironments such as viscosity, pH and polarity. Abnormal changes of mitochondrial viscosity have been shown to relate to pathological activities and diseases. Therefore, it is critical to focus our attention on mitochondrial viscosity under different conditions. A novel organic water-soluble molecule called JLQL that could monitor viscosity was conveniently synthesized in two steps. The near-infrared sensor with maximum emission wavelength of 734.6 nm and the Stokes shift of 134.6 nm consisted of a fluorophore and a mitochondrial-targeting moiety as an acceptor group; the two were connected by a double bond. The fluorescence intensity of the sensor increased 175 times with the enhancement of viscosity of a PBS-glycerol system. The interference of other microenvironments such as pH and polarity and other interference analytes could be reduced. JLQL could sensitively and selectively differentiate different levels of mitochondrial viscosity induced by monensin or nystatin. Furthermore, the probe may provide an attractive way to monitor real-time changes of viscosity during mitophagy. Possessing the above properties, JLQL can potentially be employed as a powerful tool for the observation of mitochondrial viscosity.

Ratiometric fluorescent sensor based on MoS2 QDs and AuNCs for determination and bioimaging of alkaline phosphatase.

Herein, a ratiometric fluorescent sensor based on MoS2 quantum dots (QDs) and glutathione-capped gold nanoclusters (AuNCs) was developed for determination and imaging of alkaline phosphatase (ALP). The sensor was developed by covalently linking QDs with AuNCs to form stable MoS2@AuNCs nanohybrids that exhibited the blue fluorescence of MoS2 QDs and the red fluorescence of AuNCs. In the presence of Ce3+, the fluorescence intensity of AuNCs was increased due to the aggregation-induced emission enhancement (AIEE), while that of MoS2 QDs remained unchanged, thus could be used as a reference signal. After adenosine 5'-monophosphate (AMP) and ALP were introduced into the system, AMP was hydrolyzed to adenosine and phosphate ions (PO43-). Owing to higher affinity between Ce3+ and PO43-, the AIEE effect was inhibited, in turn resulting in the decrease of AuNCs fluorescence. The developed ratiometric fluorescent sensor had a linear response to ALP concentration ranging from 0.5 to 50 U L-1 with a detection limit (LOD) of 0.08 U L-1. Moreover, the sensor had low cytotoxicity and was successfully employed in lysosome localization and bioimaging of intracellular ALP in living cells.

Reparative Dentin Formation by Dentin Matrix Proteins and Small Extracellular Vesicles.

INTRODUCTION: Vital pulp therapy aims at preserving pulp vitality and regenerating dentin. Therefore, the purpose of this study was to explore the effects of a combination of treated dentin matrix (TDM) proteins and dental pulp cell (DPC)-derived small extracellular vesicles (sEVs) on pulp-dentin complex repair. METHODS: We prepared TDM by chemical demineralization and mechanical disruption of teeth to a powder preparation. The sEVs were isolated from culture supernatants of DPCs and identified by nanoparticle tracking analysis, Western blotting, and transmission electron microscopy. The effect of a combination of TDM proteins and DPC-derived sEVs on DPC proliferation, migration, and odontogenic differentiation was evaluated in vitro. A minipig model of pulp injury was used to compare the clinical outcomes and tissue responses attributed to 4 materials including TDM, sEV-TDM, sEVs, and mineral trioxide aggregate. RESULTS: The sEV isolated from the cell supernatant promoted DPC proliferation and migration. The combination of TDM extracts and sEV synergistically promoted the migration of DPCs but suppressed their proliferation. Real-time polymerase chain reaction and Western blot revealed that sEV-TDM enhanced the odontoblast-related protein expressions in DPCs. In in vivo studies, TDM and sEV-TDM promoted the formation of continuous reparative dentin. Furthermore, odontoblastlike high columnar cells were observed on the pulp side of the dentin bridge. CONCLUSIONS: The sEV-TDM complex exhibits intrinsic biological activities, which has potential applications as a bioactive pulp-capping material.

A sensitive "off-on" carbon dots-Ag nanoparticles fluorescent probe for cysteamine detection via the inner filter effect.

In this study, we describe the construction of an "off-on" fluorescent probe based on carbon dots (CDs) and silver nanoparticles (AgNPs) mixture for sensitive and selective detection of cysteamine. By mixing AgNPs with CDs solution, the fluorescence of CDs was significantly decreased due to the inner filter effect (IFE). Upon addition of cysteamine to the mixed aqueous of CDs and AgNPs, the silver-sulfur bond between cysteamine and AgNPs caused AgNPs to aggregate, and the quenched fluorescence of CDs could in turn be recovered. The probe was employed to quantitatively detect cysteamine, and the results showed that it could detect cysteamine in a concentration range of 2-16 µM with the detection limit of 0.35 µM (signal-to-noise ratio of 3). The detection of cysteamine spiked into bovine serum samples showed high recovery rates ranging from 95.5 to 111.7%. More importantly, the developed probe had low cytotoxicity and was successfully used for in vivo imaging of HepG2 cells.

Lysosome-targeted ratiometric fluorescent sensor for monitoring pH in living cells based on one-pot-synthesized carbon dots.

A hydrothermal method has been employed to synthesize a green and one-pot carbon dots-based sensor for ratiometric monitoring and imaging lysosomal pH in living cells. The carbon dots were directly functionalized by abundant amino groups during synthesis and exhibited dual emission bands at 439 and 550 nm under single-wavelength excitation of 380 nm without any additional modification. In addition to its small size, the established sensor had good biocompatibility. Owing to its abundant amino groups and good hydrophilicity, the sensor is able to target lysosome with high Pearson's colocalization coefficients (0.935 and 0.924) and responds to change of lysosomal pH in living cells. It also had excellent pH sensitivity and reversibility, and anti-interference capability, thus enabling sensing pH change in intracellular environment in real time, as demonstrated by successful monitoring of lysosomal pH changes during lysosomal alkalization, dexamethasone-induced stimulation, and stress in Michigan Cancer Foundation-7 cells (blue channel, excitation = 405 nm and emission = 419-459 nm bandpass; and yellow channel, excitation = 405 nm and emission = 530-570 nm bandpass). Graphical abstract.

A novel near-infrared fluorescent probe for intracellular detection of cysteine.

Cysteine (Cys) takes part in redox balance in cells as an antioxidant, and imbalance of Cys content in the body can cause a variety of diseases. It is very important to develop a new fluorescent chemosensor to specifically detect Cys intracellular. In this work, a novel NIR fluorescent probe was constructed based on 3-ethyl-1,1,2-trimethyl-1H-benzo[e]indol-3-ium iodide and 5-(4-hydroxyphenyl)furan-2-carbaldehyde. The probe could selectively detect Cys in the presence of homocysteine (Hcy), glutathione (GSH), and other interferences. It also had a number of advantages, including nucleolus-targeting ability, long fluorescence emission wavelength (685 nm), low detection limit (56 nM), and large Stokes shift (172 nm). The probe was employed to enable visualization of Cys in HepG2 cells, and due to its good response in viscous environment, the probe could also locate nucleoli intracellular.