A reversible CHEF-based NIR fluorescent probe for sensing Hg2+ and its multiple application in environmental media and biological systems.

Hg2+ poses a great threat to human health and the environment due to its bioaccumulation and permanent damage. Herein, a reversible CHEF-based near-infrared fluorescent probe 2-(3-((E)-4-((E)-4-(diethylamino)-2- hydroxybenzylidene)amino)styryl)-5,5-dimethylcyclohex-2-en-1-ylidene)propanedinitrile (DHEY) capable of specifically recognizing Hg2+ was constructed. DHEY exhibits advantages of large Stokes shift (157 nm), excellent selectivity, high sensitivity (LOD = 3.2 µg/L), and fast response efficiency (<3 min). Interestingly, DHEY can also realize rapid and effective detection of Hg2+ after being recycled 7 times. The successful recovery of trace Hg2+ in different environmental water samples fully demonstrates the potential of DHEY for actual applications. In particular, DHEY enables real-time observation of the distribution and translocation pattern of exogenous Hg2+ in HeLa cells and zebrafish. This work provides important theoretical support for investigating the fate of heavy metal ions in the environment using fluorescence techniques.

An ICT-Based Coumarin Fluorescent Probe for the Detection of Hydrazine and Its Application in Environmental Water Samples and Organisms.

As an inorganic small molecule pollutant, the toxicity and potential carcinogenicity of hydrazine (N2H4) are of increasing concern. In this work, A water-soluble fluorescent probe (OCYB) based on the intramolecular charge transfer (ICT) mechanism for the detection of hydrazine was designed and synthesized. Taking the advantage of 4-bromobutyryl as the recognition group, the high selectivity of OCYB to N2H4 was confirmed by steady-state fluorescence spectroscopy. The limit of detection (LOD) was calculated to be 78 nM in the DMSO-HEPES (pH 7.4) system. The detection mechanism was verified by NMR, HRMS and density functional theory (DFT) calculations. In addition, OCYB exhibits strong anti-interference ability and an "Off-On" fluorescence enhancement effect. Importantly, OCYB can be used to effectively monitor the fluorescence distribution of N2H4 in environmental water samples and organisms.

A novel ESIPT-based fluorescent probe with dual recognition sites for the detection of hydrazine in the environmental water samples and in-vivo bioimaging.

Hydrazine (N2H4), an important chemical intermediate, has been widely used in industrial production and agricultural life, but it has also caused environmental pollution. A novel ESIPT-based fluorescent probe with dual recognition sites, 2-(benzothiazole-2-yl)-1,4-imphenyl bis 4-bromobutyric acid (BRBA), was developed to selectively detect N2H4 under complex conditions. BRBA exhibits accurate detection for N2H4 with a good linear relationship ranging from 0 to 150 µM, and the LOD can reach 0.1 µM. Importantly, taking advantage of low cytotoxicity and a large Stokes shift, BRBA can be utilized to monitor environmental water samples and successfully applied to imaging HeLa cells and zebrafish.

Pulsatilla decoction alleviates colitis by enhancing autophagy and regulating PI3K­Akt­mTORC1 signaling pathway.

The aim of the present study was to investigate the therapeutic effect of Pulsatilla decoction (PD) on ulcerative colitis (UC) and to elucidate its potential molecular mechanisms. C57BL/6 mice expressing natural killer (NK)1.1 were used as experimental animals in the present study and a model of oxazolone­induced colitis was established. Mice were randomly divided into the following five groups: i) PD group; ii) oxazolone­induced colitis group; iii) IL­13 intervention group; iv) 5­aminosalicylic acid positive control group; and v) negative control group (equal volume saline gavage). A total of 10 animals were used in each group. The effects of PD on UC and the association between this regimen and the PI3K­Akt­mTORC1 signaling pathway were evaluated by disease activity index (DAI), hematoxylin and eosin staining, reverse transcription­quantitative PCR (RT­qPCR), immunofluorescence assay, ELISA and western blotting. The UC models were successfully established by injecting oxazolone gavage solution. Clinical colitis evaluation and histological examination revealed that PD reduced the DAI values in oxazolone­induced colitis in mice and the degree of infiltration in NK1.1 cells. PD significantly reduced the secretion of IL­13, as determined using an ELISA. In addition, western blotting and RT­qPCR analyses demonstrated that Beclin1 and LC3II/I expression levels were downregulated following treatment of the mice with PD. In addition, PD not only partially restored alterations in the expression of tight junction proteins in the colon tissues, but also suppressed the activation of the PI3K­Akt­mTORC1 signaling pathway. The data indicated that this regimen could alleviate oxazolone­induced UC in mice, which could significantly reduce tissue inflammation and autophagy. The mechanism of action was associated with the PI3K­Akt­mTORC1 signaling pathway.

New insights into the interactions between humic acid and three neonicotinoid pesticides, with multiple spectroscopy technologies, two-dimensional correlation spectroscopy analysis and density functional theory.

The widespread use of neonicotinoid pesticides in agricultural production has caused pressure on the environment. In the present work, the interactions between humic acid (HA) and three neonicotinoid insecticides, dinotefuran, clothianidin and nitenpyram, were investigated by using multiple spectroscopy techniques combined with two-dimensional correlation spectroscopy analysis and density functional theory (DFT). Dinotefuran, clothianidin and nitenpyram could quench the endogenous fluorescence of HA through a static quenching process dominated by hydrogen bonds and van der Waals forces. According to the revised Stern-Volmer equation and DFT calculation, the binding abilities of the three pesticides with HA were ranked as dinotefuran < clothianidin < nitenpyram. The results of dynamic light scattering showed that neutral conditions were more conducive to the combination of HA and dinotefuran, clothianidin and nitenpyram. Through Fourier transform infrared spectroscopy (FTIR) combined with two-dimensional correlation analysis (2D-COS), the functional group with the strongest binding ability in the HA-dinotefuran, HA-clothianidin and HA-nitenpyram system was CH, CO and CO, respectively. The work will help to further understand the behavior of neonicotinoid pesticides in the environment.

Hydrazine exposure: A near-infrared ICT-based fluorescent probe and its application in bioimaging and sewage analysis.

Hydrazine (N2H4) is an environment pollutant with high acute toxicity and potential carcinogenicity, and detection of N2H4 has attracted increasing attention. In the present study, a low toxicity near-infrared fluorescent probe (DCDB) based on the intramolecular charge transfer (ICT) principle was developed. The probe DCDB exhibits excellent selectivity and high sensitivity (LOD = 1.27 ppb) for N2H4, fast reaction rate (5 min), extremely large Stokes shift (160 nm). The color transformation of the DCDB-N2H4 system from purple to pink can be observed with the naked eye. The success of N2H4 test strips to detect trace N2H4 in actual sewage strongly illustrates the practical application potential of DCDB. Importantly, DCDB can be utilized to monitor the distribution of exogenous N2H4 in vivo and in vitro.

A novel near-infrared fluorimetric method for point-of-care monitoring of Fe2+ and its application in bioimaging.

Iron is one of the essential trace elements in the human body, which is involved in many important physiological processes of life. The abnormal amount of iron in the body will bring many diseases. Therefore, a novel near-infrared fluorimetric method was developed. The method is based on a fluorescent probe (E)-4-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohex-1-en-1-yl)vinyl)-N, N-diethylaniline oxide (DDED) which uses N-oxide as a recognition group to real-time monitoring and imaging of Fe2+ in vivo and in vitro. The method exhibits excellent selectivity and high sensitivity (LOD = 27 nM) for Fe2+, fast reaction rate (< 4 min), extremely large Stokes shift (＞ 275 nm), low cytotoxicity. The strip test strongly illustrates the potential application of DDED in real environment. In particular, DDED has been successfully applied to real-time monitoring and imaging of Fe2+ in HepG2 cells and zebrafish. That is, the method has great potential for the detection of Fe2+ in living systems.

Interactions between Imidacloprid and Thiamethoxam and Dissolved Organic Matter Characterized by Two-Dimensional Correlation Spectroscopy Analysis, Molecular Modeling, and Density Functional Theory Calculations.

The heavy application of neonicotinoid insecticides in agricultural production has burdened the environment. In the present study, interactions of two neonicotinoid insecticides imidacloprid and thiamethoxam with dissolved organic matter (DOM) were investigated by spectroscopic techniques, molecular modeling, and density functional theory (DFT) calculations. The static mechanism of imidacloprid and thiamethoxam quenching the endogenous fluorescence of DOM was assessed through time-resolved analyses. During the binding process, a protein-like substance binds imidacloprid and thiamethoxam later than a humic-like substance, as analyzed by two-dimensional correlation spectroscopy, but more strongly than the humic-like substance, as suggested by molecular modeling and DFT calculations. The conformational changes of DOM are attributed to imidacloprid and thiamethoxam, as assessed with three-dimensional spectra. Fourier transform infrared spectroscopy indicated that DOM binds imidacloprid and thiamethoxam by hydroxyl, aliphatic C-H, amide I, and carboxyl to form stable DOM-imidacloprid and DOM-thiamethoxam complexes. Understanding the changes in the structural conformation of humic-like and protein-like substances with imidacloprid and thiamethoxam helps further understand the fate of the neonicotinoids in the environment.

An ICT-based fluorescent probe with a large Stokes shift for measuring hydrazine in biological and water samples.

As a strong reductant and highly active alkali, hydrazine (N2H4) has been widely used in chemical industry, pharmaceutical manufacturing and agricultural production. However, its high acute toxicity poses a threat to ecosystem and human health. In the present study, a ratiometric fluorescent probe for the detection of N2H4 was designed, utilizing dicyanoisophorone as the fluorescent group and 4-bromobutyryl moiety as the recognition site. 4-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohex-1-enyl) phenyl 4-brobutanoate (DDPB) was readily synthesized and could specially sense N2H4 via an intramolecular charge transfer (ICT) pathway. The cyclization cleavage reaction of N2H4 with a 4-bromobutyryl group released phenolic hydroxyl group and reversed the ICT process between hydroxy group and fluorophore, turning on the fluorescence in the DDPB-N2H4 complexes. DDPB exhibits a low cytotoxicity, reasonable cell permeability, a large Stokes shift (186 nm) and a low detection limit (86.3 nM). The quantitative determination of environmental water systems and the visualization fluorescence of DDPB test strips provides a strong evidence for the applications of DDPB. In addition, DDPB is suitable for the fluorescence imaging of exogenous N2H4 in HeLa cells and zebrafish.

Modified Pulsatilla decoction attenuates oxazolone-induced colitis in mice through suppression of inflammation and epithelial barrier disruption.

Inflammatory bowel diseases (IBDs) are chronic inflammatory gastrointestinal disorders caused by a dysregulated mucosal immune response and epithelial barrier disruption. Conventional treatment of IBD is currently limited to overcoming patient symptoms and is often associated with severe adverse effects from the drugs used. Modified Pulsatilla decoction has been used previously to treat ulcerative colitis (UC) in clinical practice in China, however, the underlying mechanism in the treatment of UC remains to be elucidated. In the present study, the efficiency and mechanisms of modified Pulsatilla decoction in the treatment of oxazolone­induced colitis were investigated. Assessment of clinical colitis and histological examination found that the administration of modified Pulsatilla decoction attenuated the severity of oxazolone­induced colitis in mice. Measurement of cytokine concentration, western blotting and reverse transcription­quantitative polymerase chain reaction demonstrated modified Pulsatilla decoction treatment significantly reduced the secretion of pro­inflammatory cytokines and restored alterations in tight junction proteins in the colon tissues. In addition, modified Pulsatilla decoction suppressed the activation of the nuclear factor­κB signaling pathway. Thus, the findings of the present study demonstrated that modified Pulsatilla decoction offers an effective therapeutic approach for the treatment of IBD and revealed the underlying mechanisms of action offered by modified Pulsatilla decoction.

Glycyrrhizic acid attenuates CCl4-induced hepatocyte apoptosis in rats via a p53-mediated pathway.

AIM: To investigate the effect of glycyrrhizic acid (GA) on carbon tetrachloride (CCl4)-induced hepatocyte apoptosis in rats via a p53-dependent mitochondrial pathway. METHODS: Forty-five male Sprague-Dawley rats were randomly and equally divided into three groups, the control group, the CCl4 group, and the GA treatment group. To induce liver fibrosis in this model, rats were given a subcutaneous injection of a 40% solution of CCl4 in olive oil at a dose of 0.3 mL/100 g body weight biweekly for 8 wk, while controls received the same isovolumetric dose of olive oil by hypodermic injection, with an initial double-dose injection. In the GA group, rats were also treated with a 40% solution of CCl4 plus 0.2% GA solution in double distilled water by the intraperitoneal injection of 3 mL per rat three times a week from the first week following previously published methods, with modifications. Controls were given the same isovolumetric dose of double distilled water. Liver function parameters, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen fibers were evaluated by Sirius red staining. Hepatocyte apoptosis was investigated using the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) assay and the cleaved caspase-3 immunohistochemistry assay. The expression levels of p53 and apoptosis-related proteins were evaluated by immunohistochemistry or Western blotting analysis. RESULTS: After 8 wk of treatment, GA significantly reduced serum activity of ALT (from 526.7 ± 57.2 to 342 ± 44.8, P < 0.05) and AST (from 640 ± 33.7 to 462.8 ± 30.6, P < 0.05), attenuated the changes in liver histopathology and reduced the staging score (from 3.53 ± 0.74 to 3.00 ± 0.76, P < 0.05) in CCl4-treated rats. GA markedly reduced the positive area of Sirius red and the ratio of the hepatic fibrotic region (from 7.87% ± 0.66% to 3.68% ± 0.32%, P < 0.05) compared with the CCl4 group. GA also decreased the expression level of cleaved caspase-3 compared to the CCl4 group. TUNEL assay indicated that GA significantly diminished the number of TUNEL-positive cells compared with the CCl4 group (P < 0.05). GA treatment clearly decreased the level of p53 (P < 0.05) detected by immunohistochemistry and Western blotting analysis. Compared with the CCl4 group, we also found that GA reduced the Bax/Bcl-2 ratio (P < 0.05), the expression of cleaved caspase-3 (P < 0.05), cleaved caspase-9 (P < 0.05), and inhibited cytochrome C and second mitochondria-derived activator of caspases (Smac) release from mitochondria to cytoplasm, i.e., GA reduced the expression level of Smac, which inhibited c-IAP1 activity (P < 0.05), ultimately inhibiting the activity of caspase-3, according to Western blotting analysis. As a result, GA suppressed activation of the caspase cascades and prevented hepatocyte apoptosis. CONCLUSION: GA can inhibit CCl4-induced hepatocyte apoptosis via a p53-dependent mitochondrial pathway to retard the progress of liver fibrosis in rats.