Activatable fluorescent probes for early diagnosis and evaluation of liver injury.

With the increase in people's living standards, the number of patients suffering from liver injury keeps on increasing. Traditional diagnostic methods can no longer meet the needs of early and accurate diagnosis due to their limitations in application. However, fluorescent probes based on different fluorophores and nanomaterials have been gradually lighting up medical research due to their unique properties, such as high specificity and non-invasiveness. In addition, accurate identification of the different types of liver injury biomarkers can significantly improve the level of early diagnosis. Therefore, this review reviews the fluorescent probes used in the detection of biomarkers of liver injury over recent years and briefly summarizes the corresponding biomarkers of different types of liver injury. Impressively, this review also lists the structures and the response mechanisms of the different probes, and concludes with an outlook, suggesting directions in which improvements can be made. Finally, we hope that this review will contribute to the further development of fluorescent probes for the early diagnosis and assessment of liver injury.

The Application of Hydrogen Sulfide Fluorescent Probe in Food Preservation, Detection and Evaluation.

This work primarily reviewed the response mechanism of fluorescent probes for H2S detection in foodstuffs in recent years, as well as the methodologies employed for detecting foodstuffs. Firstly, the significance of studying H2S gas as an important signaling molecule is introduced. Subsequently, a review of the response mechanism of the scientific community on how to detect H2S in foodstuffs samples by fluorescent probe technology is carried out. Secondly, the methods commonly used for detecting foodstuffs samples are discussed, including the test strip method and the spiking recovery methods. Nevertheless, despite the significant advancements in this field, there remain some research gaps. Finally, the article identifies the remaining issues that require further attention in current research and proposes avenues for future investigation. More importantly, this work identifies the current limitations of research in this field and proposes future applications of fluorescent probes for H2S in assessing food freshness and determining food spoilage. Therefore, this review will provide robust technical support for the protection of consumer health and the advancement of the sustainable development of the food industry and also put forward some new ideas and suggestions for future research.

Visible-light-promoted trifluoromethylselenolation of ortho-hydroxyarylenaminones.

Development of an efficient process that employs easy to handle and shelf-stable reagents for the synthesis of trifluoromethylselenylated heterocyclics remains a daunting challenge in organic synthesis. Herein, we report a green and practical protocol using trifluoromethyl tolueneselenosulfonate and ortho-hydroxyarylenaminones to access a wide range of chromone derivatives under photocatalyst and oxidant free conditions. This reaction proceeded smoothly under photoirradiation conditions and various functional groups were tolerant of the reaction conditions.

A Review for In Vitro and In Vivo Detection and Imaging of Gaseous Signal Molecule Carbon Monoxide by Fluorescent Probes.

Carbon monoxide (CO) is a vital endogenous gaseous transmitter molecule involved in the regulation of various physiological and pathological processes in living biosystems. In order to investigate the biological function of CO, many technologies have been developed to monitor the level of endogenous CO in biosystems. Among them, the fluorescence detection technology based on the fluorescent probe has the advantages of high sensitivity, excellent selectivity, simple operation, especially non-invasive damage to biological samples, and the possibility of real-time in situ detection, etc., which is considered to be one of the most effective and applicable detection techniques. Therefore, in the last few years, a lot of work has been carried out on the design, synthesis and in vivo fluorescence imaging studies of CO fluorescent probes. Furthermore, using fluorescent probes to detect the changes in CO concentrations in living cells and tissues as well as in organisms has been one of the hot research topics in recent years. However, it is still a challenge to rationally design CO fluorescent probe with excellent optical performance, structural stability, low background interference, good biocompatibility, and excellent water solubility. Therefore, this review focuses on the research progress of CO fluorescent probes in the detection mechanism and biological applications in recent years. However, this popular and leading topic has rarely been summarized comprehensively to date. Thus, the research progress of CO fluorescent probes in recent years is reviewed in terms of their design concept, detection mechanism, and their biological applications. In addition, the relationship between the structure and performance of the probes was also discussed. More significantly, we hope that more excellent optical properties fluorescent probes for gaseous transmitter molecule CO detection and imaging will overcome the current problems of high biotoxicity and limited water solubility in future.

Selective difunctionalization of electron-deficient alkynes: access to (E)-2-iodo-3-(methylthio)acrylate.

A convenient and efficient approach to (E)-2-iodo-3-(methylthio)acrylate has been developed through direct iodothiomethylation of alkynes with aqueous HI and DMSO under mild conditions. This novel protocol has demonstrated a unique difunctionalization of electron-deficient alkynes with a broad substrate scope and excellent functional-group tolerance. Preliminary mechanistic studies indicated that prior diiodination of alkynes, followed by nucleophilic substitution with in situ generated DMS led to the formation of (E)-2-iodo-3-(methylthio)acrylate.

Design, synthesis and biological Evaluation of Dual acetyl cholinesterase and beta-secretase inhibitors in treatment for alzheimer's Disease.

With the recent research advances in molecular biology and technology multiple credible hypotheses about the progress of Alzheimer's disease (AD) have been proposed, among which the amyloid and cholinergic hypotheses are commonly used to develop reliable therapeutic agents. The multitarget-directed ligand (MTDL) approach was taken in this work to develop muilti-functional agents, which can mainly serve as dual beta-secretase (BACE 1) and Acetylcholinesterase (AChE) inhibitors. Series of new compounds were designed, synthesized and evaluated in this work, from which we identified 2-((4-(1,3-dioxoisoindolin-2-yl)benzyl)amino)-2-oxoethyl-2-(4-methoxyphenyl)acetate (1h) as a new dual cholinesterase and beta-secretase inhibitor without toxicity.

Design, synthesis and biological evaluation of dual acetylcholinesterase and phosphodiesterase 5A inhibitors in treatment for Alzheimer's disease.

With the recent research advances in molecular biology and technology, multiple credible hypotheses about the progress of Alzheimer's disease (AD) have been proposed; multi-target drugs have emerged as an innovative therapeutic approach for AD. Current clinical therapy for AD patients is mainly palliative treatment targeting acetylcholinesterase (AChE). Inhibition of phosphodiesterase 5A (PDE5A) has recently been validated as a potentially novel therapeutic approach for Alzheimer's disease (AD). In this work, series of new compounds were designed, synthesized and evaluated as dual cholinesterase and PDE5A inhibitor. Biological results revealed that some of these compounds display good biological activities against AChE with IC50 values about 44.67-169.80nM (donepezil IC50 50.12nM). Notably, compound 12 presented potent activities against PDE5A with IC50 values about 50µM (sildenafil IC50 12.59µM), and some of these compounds showed low cell toxicity to A549 cells in vitro.

ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging.

Photoactivatable fluorophores have become an important technique for the high spatiotemporal resolution of biological imaging. Here, we developed a novel photoactivatable probe (PHBT), which is based on 2-(2-hydroxyphenyl)benzothiazole (HBT), a small organic fluorophore known for its classic luminescence mechanism through excited-state intramolecular proton transfer (ESIPT) with the keto form and the enol form. After photocleavage, PHBT released a ratiometric fluorophore HBT, which showed dual emission bands with more than 73-fold fluorescence enhancement at 512 nm in buffer and more than 69-fold enhancement at 452 nm in bovine serum. The probe displayed a high ratiometric imaging resolution and is believed to have a wide application in biological imaging.

A New Two-Photon Ratiometric Fluorescent Probe for Detecting Alkaline Phosphatase in Living Cells.

Alkaline phosphatase (ALP) is an important diagnostic indicator of many human diseases. To quantitatively track ALP in biosystems, herein, for the first time, we report an efficient two-photon ratiometric fluorescent probe, termed probe 1 and based on classic naphthalene derivatives with a donor-π-acceptor (D-π-A) structure and deprotection of the phosphoric acid moiety by ALP. The presence of ALP causes the cleave of the phosphate group from naphthalene derivatives and the phosphate group changes the ability of the intramolecular charge transfer (ICT) and remarkably alters the probe's photophysical properties, thus an obvious ratiometric signal with an isoemissive point is observed. The fluorescence intensity ratio displayed a linear relationship against the concentration of ALP in the concentration range from 20 to 180 U/L with the limit of detection of 2.3 U/L. Additionally, the probe 1 is further used for fluorescence imaging of ALP in living cells under one-photon excitation (405 nm) or two-photon excitation (720 nm), which showed a high resolution imaging, thus demonstrating its practical application in biological systems.

A high-fidelity dual-channel fluorescence probe for benzoyl peroxide detection and toxicity early warning in food, zebrafish, and mice.

Benzoyl peroxide (BPO), as a widely used organic peroxide, has attracted widespread attention from all sectors of society for its environmental hazards and potential risks to human health. Herein, we employed a Förster resonance energy transfer (FRET) strategy to construct a novel ratiometric fluorescent probe CY-DCI for BPO detection in food, zebrafish, and mice. Specifically, a hemicyanine fluorophore and a dicyanoisophorone fluorophore were connected with a piperazine group as donor and acceptor, respectively, and an olefinic unsaturated bond as the reaction site. CY-DCI has favorable selectivity and an excellent detection limit as low as 58.1 nM, and the recovery rates for real-sample detection ranged from 95.8 % to 104 %, with relative standard deviations (RSD) less than 2.58 %. To further improve its practicality, silica gel plates and test strips containing CY-DCI (0-50 µM) were developed for naked-eye detection of BPO with satisfactory results. Additionally, this novel probe was then applied for ratiometric imaging of living zebrafish and mice and showed high ratiometric imaging resolution in the green and red channels, thus demonstrating its practical application for BPO detection and toxicity early warning in food and biosystems.

Construction of a super large Stokes shift near-infrared fluorescent probe for detection and imaging of superoxide anion in living cells, zebrafish and mice.

As one of the major reactive oxygen species (ROS), superoxide anion (O2•-) is engaged in maintaining redox homeostasis in the cell microenvironment. To identify the pathological roles in related disorders caused by abnormal expression of O2•-, it is of great significance to monitor and track the fluctuation of O2•- concentration in vivo. However, the low concentration of O2•- and the interference caused by tissue autofluorescence make the development of an ideal detection methodology full of challenges. Herein, a "Turn-On" chemical response near-infrared (NIR) fluorescence probe Dcm-Cu-OTf for O2•- detection in inflamed models, was constructed by conjugating the NIR fluorophore (dicyanisophorone derivative) with an O2•- sensing moiety (trifluoromethanesulfonate). Dcm-Cu-OTf exerted about 140-fold fluorescence enhancement after reacting 200 µM O2•- with an excellent limited of detection (LOD) as low as 149 nM. Additionally, Dcm-Cu-OTf exhibited a super large Stokes shift (260 nm) and high selectivity over other bio-analytes in stimulated conditions. Importantly, Dcm-Cu-OTf showed low toxicity and enabled imaging of the generation of O2•- in the Lipopolysaccharide (LPS)-stimulated HeLa cells, zebrafish, and LPS-induced inflamed mice. The present study provided a potential and reliable detection tool to inspect the physiological and pathological progress of O2•- in living biosystems.

A TP-FRET-based fluorescent sensor for ratiometric visualization of selenocysteine derivatives in living cells, tissues and zebrafish.

Selenium is a biologically essential micronutrient element serving as an essential building block for selenoproteins (SePs), which is playing a key role in various cellular functions. Hence, it is of great significance to developing a reliable and rapid method for detection of Sec in biosystems. Compared with the previously reported probes that have been developed for selective detection of Sec, two-photon (TP) ratiometric Sec-specific probes would be advantageous for the NIR excitation and built-in correction of the dual emission bands. To quantitatively and selectively detect Sec over biothiols with rapid and sensitive response, we for the first time report a new fluorescence resonance energy transfer (FRET)-based TP ratiometric fluorescence probe CmNp-Sec, which was constructed by conjugating a TP fluorophore 6 (coumarin derivative with a D-π-A-structure) with a naphthalimide fluorophore 9 via a non-conjugated linker, and employed a 4-dinitrobenzene-ether (DNB) with a strong ICT effect as Sec responsive moiety. It exhibits quantitatively detect Sec in a wide range (0-50 µM) with a limit of detection of 7.88 nM within 10 min. More impressively, this probe can be conveniently used to detect Sec in living cells, tissues and zebrafish, demonstrating it has the latent capability in further biological applications.

A novel "turn-on" mitochondria-targeting near-infrared fluorescent probe for H2S detection and in living cells imaging.

Hydrogen sulfide (H2S) has been considered to be involved in cytoprotective processes and redox signaling. It is very meaningful to track and analyze it in mitochondria. Herein, we report a novel "turn-on" mitochondria-targeting near-infrared fluorescent probe (Mito-NIR-SH) for detection of H2S in living cells, which was designed and synthesized by introducing 2,4-dinitrophenyl as fluorescence quenching group and H2S response moiety into Changsha near-infrared fluorophore (CS-OH). The structure of the fluorophore and the probe were characterized by 1H NMR, 13C NMR and mass spectrometry. Meanwhile, Mito-NIR-SH could quantitatively detect H2S at concentrations ranging from 0 to 30 µM with a detection limit as low as 89.3 nM, showing good chemical stability, fast "turn-on" response, selectively mitochondrial location, as well as high sensitivity and selectivity toward H2S. Based on this, it was successfully applied to imaging exogenous and endogenous H2S in living HeLa cells via confocal fluorescence microscopy.

A novel "turn-on" mitochondria-targeting near-infrared fluorescent probe for determination and bioimaging cellular hydrogen sulfide.

Hydrogen sulfide (H2S) has been regarded as an important gas transmitter playing vital role in cytoprotective processes and redox signaling. It is very meaningful to monitor and analyze it in biosystem for obtaining important physiological and pathological information. Despite numerous fluorescent probes for cellular H2S have been reported in past decades, only a few have capability to detect mitochondrial H2S with near-infrared (NIR) emission. Therefore, a new mitochondria-targeting NIR fluorescent probe (Mito-NSH) for detection of cellular H2S was developed by introducing 2,4-dinitrophenyl ether into a novel dye (Mito-NOH). A large "turn-on" NIR fluorescence response was obtained due to thiolysis of ether to hydroxyl group when Mito-NSH was treated with NaHS. Moreover, Mito-NSH could quantitatively detect H2S at concentration ranging from 0 to 30 µM with a detection limit of 68.2 nM, and it exerts some superior optical properties, such as large stokes shift (107 nm), highly selectively mitochondria location, fast response and high selectivity to H2S. More impressively, it was successfully applied to imaging exogenous and endogenously generated H2S in living HeLa cells via confocal fluorescence microscopy.

A new "off-on" NIR fluorescence probe for determination and bio-imaging of mitochondrial hypochlorite in living cells and zebrafish.

Hypochlorite anion (ClO-) has been recognized as host defense destructing incursive bacteria and pathogens, a signal molecule inducing occurrence of apoptosis and a noxious agent when it is overproduced. It is significant to detect ClO- in mitochondria for getting meaningful physiological and pathological information. Compared with the fluorescence probes of emission wavelength in ultraviolet or visible region, those with near-infrared (NIR) fluorescence signal are advantageous due to the deeper tissue penetrability and less photo-bleaching effect. In this work, a new "off-on" NIR ClO--specific fluorescence probe (Mito-NClO) especially located in mitochondria was designed and synthesized by condensation of diaminomaleonitrile with a new fluorophore (Mito-NCHO). A marked "turn-on" NIR fluorescence signal was observed on account of the oxidation of the imine bond by NaClO. Moreover, in the range from 0 to 20 µM, this probe had the capability to quantitatively detect ClO- with a detection limit as low as 90.2 nM. Additionally, the probe exerted other excellent properties, including larger stokes shift (117 nm), better aqueous solubility, high selectivity toward ClO-, rapid response and selective mitochondrial location. Furthermore, the bio-imaging experiments clearly demonstrated that Mito-NClO facilitated the visualization of exogenous and endogenous ClO- in living HeLa cells and zebrafish model. Therefore, we speculate that the probe Mito-NClO can be served as an ideal tool for the monitoring of ClO- in biosystems.

Paper-based microfluidic devices for electrochemical immunofiltration analysis of human chorionic gonadotropin.

An electrochemical immunofiltration analysis was introduced into microfluidic paper-based analytical devices (µPADs) for the first time, which was based on photolithography and screen-printing technology. The hydrophilic test zones of the aldehyde-functionalized screen-printed electrodes (SPEs) were biofunctionalized with capture antibodies (Ab1). A sensitive immune detection method was developed by using primary signal antibody functionalized gold nanoparticles (GNPs/Ab2) and alkaline phosphatase conjugated secondary antibody (ALP-IgG). Differential pulse voltammetry (DPV) was performed to detect the electrochemical response. The microfluidic paper-based electrochemical immunosensor (µ-PEI) was optimized and characterized for the detection of human chorionic gonadotropin (HCG), a model analyte, in a linear range from 1.0mIUmL-1 to 100.0 IU mL-1 with a detection limit of 0.36mIUmL-1. Additionally, the proposed µ-PEI was used to test HCG in real human serum and obtained satisfactory results. The disposable, efficient, sensitive and low-cost µ-PEI has exhibited great potential for the development of point-of-care testing (POCT) devices that can be applicated in healthcare monitoring.

Multipotent AChE and BACE-1 inhibitors for the treatment of Alzheimer's disease: Design, synthesis and bio-analysis of 7-amino-1,4-dihydro-2H-isoquilin-3-one derivates.

In this paper, the preparation of a new class of multi-target-directed ligands (MTDLs) based on a 7-amino-1,4-dihydro-2H-isoquilin-3-one, whose lead (compound I) showed promising properties in acetylcholinesterase (AChE) inhibitory activity [1], is described. The results of in vitro activities and molecular docking demonstrated that the target molecule (compounds 10a-n) with three parts of aromatic moieties and appropriate structural length can interact with aromatic residues in catalytic active site (CAS), peripheral anionic site (PAS) and the channel of AChE. And the introduce of connecting amide bonds, enables the target molecules provide sufficient hydrogen bond donors and acceptors to interact with the catalytic site of BACE-1. Notably, compound 10d exerted excellent AChE inhibition (IC50 = 18.93 ± 1.02 pM, 181-fold more inhibitory effect compared with donepezil), BACE-1 inhibition (97.68 ± 8.01% at 20 µM), and good metal chelating property, which can be chosen as lead compound for further optimization of novel small ligand for the treatment of Alzheimer's disease.

A naphthalene-based two-photon fluorescent probe for selective and sensitive detection of endogenous hypochlorous acid.

An efficient naphthalene-based two-photon fluorescent probe for endogenous HClO has been reported in the present study, which consists of a 6-(2-benzothiazolyl)-2-naphthalenol fluorophore connected with a 4-aminophenol (the fluorescence quenching and response group). This probe exhibits a high selectivity and excellent sensitivity with a detection limit of 7.6nM over other reactive oxygen species and analyte species, and the fluorescence intensity enhanced 103-fold when responsed. Furthermore, it was successfully used for two-photon imaging of endogenous HClO in live cells with high-resolution.

Chelerythrine and Fe3O4 Loaded Multi-Walled Carbon Nanotubes for Targeted Cancer Therapy.

The work focused on manufacturing improved drug loaded multifunctional magnetic nanoparticles that can overcome the relative non-specificity and potential side-effects of some chemotherapeutic drugs to healthy tissues. A new drug delivery system, Chelerythrine (CHE) and Fe3O4 loaded multi-walled carbon nanotubes (Fe3O4/MWNTs-CHE nanocomposites) that can target hepatocytes when treating malignant tumors, was prepared through a simple adsorption method. The formulation and structure of the Fe3O4/MWNTs-CHE nanocomposites were characterized by vibrating sample magnetometer (VSM), Fourier Transform infrared spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The cytotoxicity and anti-proliferation effect from the prepared nanocomposites were in vitro tested on human hepatocarcinoma HepG2 and normal liver LO2 cell lines. The results showed the saturated magnetization of Fe3O4/MWNTs-CHE nanocomposites could reach to 45.4O3 emu/g, and the in vitro CHE release behavior exhibited a biphasic release pattern. Moreover, the in vitro cytotoxicity studies revealed that the Fe3O4/MWNTs-CHE nanocomposites showed an efficient inhibition rate to HepG2 cell line and exhibited a lower cytotoxicity to LO2 cell line in comparison to the native CHE. Therefore, the multifunctional Fe3O4/MWNTs-CHE nanocomposites should be a useful and promising candidate for treatment of malignant tumors.

Aptasensors application for cow's milk allergens detection and early warning: Progress, challenge, and perspective.

Cow's milk allergy (CMA) is considered one of the most prevalent food allergies and a public health concern. Modern medical research shows that the effective way to prevent allergic reactions is to prevent allergic patients from consuming allergenic substances. Therefore, the development of rapid and accurate detection technology for milk allergens detection and early warning is critical to safeguarding those with a cow milk allergy. As the oligonucleotide sequences with high specificity and selectivity, aptamers frequently assemble with transduction elements forming multifarious aptasensors for quantitative detection owing to their high-affinity binding to the target. Current aptasensors in the field of cow's milk allergen detection in recent years are explored in this review. This review takes a look back at a few common assays, including ELISA and PCR, before presenting a clear overview of the aptamer and threshold doses. It delves into a detailed discussion of the current aptamer-based detection techniques and related theories for milk allergen identification. Last but not least, we conclude with a discussion and outlook of the advancements made in allergen detection with aptamers. We sincerely hope that there will be more extensive applications for aptasensors in the future contributing to reducing the possibility of patients suffering from adverse reactions.