Fluorimetric Tool to Discriminate Glomerular and Tubular Injuries In Vivo.

The etiology and pathological complexity of acute kidney injury (AKI) pose great challenges for early diagnosis, typing, and personalized treatment. It is an important reason for poor prognosis and high mortality of AKI. In order to provide a relatively noninvasive diagnostic and typing method for AKI, we proposed the pathological changes of albumin permeability after glomerular injury and reabsorption efficiency after tubular injury as potential entry points. Thus, a renal tubule labeling fluorescent dye which features albumin concentration-related fluorescence intensity was used to fit these pathological changes. Utilizing this fluorescence assay, we realized urinary tract obstruction imaging as early as 12 h after morbidity. For glomerular and tubular injury discrimination, compared to a healthy control, membranous nephropathy as a representative glomerular injury resulted in enhanced fluorescence intensity of the kidney due to increased albumin penetration, while renal tubular injury caused insufficient dye reabsorption to exhibit weakened fluorescence intensity. The significant differences demonstrated the feasibility of this approach for fluorescence imaging-based AKI typing in vivo.

A Case of Severe Lethal Allergic Reaction Caused by Iodixanol After Digital Subtraction Angiography.

BACKGROUND: Severe lethal allergic reactions triggered by iodixanol following digital subtraction angiography (DSA) are rare. The majority of skin reactions associated with iodixanol were mild, and the prognosis was favorable. Moreover, a case of serious skin adverse events caused by iodixanol has been documented. METHODS: A 61-year-old woman underwent surgery for a cerebral hemorrhage in another hospital. Upon the surgery, the patient's state of impaired consciousness did not show any improvement. Head computed tomography angiography on admission: right middle cerebral artery M1 segment enlargement, left posterior cerebral artery P2 stenosis. Following undergoing DSA with iodixanol, the patient experienced severe and fatal drug eruptions, which represents a serious and uncommon complication associated with iodixanol. RESULTS: This paper describes the experience in the treatment and nursing of severe allergic reactions. Despite the fact that the patient was discharged automatically and eventually died, there are valuable lessons to be learned from this case that can inform and guide future clinical practices. CONCLUSIONS: Iodixanol adverse reactions were rare, and severe fatal adverse reactions were seldom reported. Consequently, the authors conclude that the potential adverse reaction risk of iodixanol contrast agent should be taken into consideration in future endeavors, and the skin and allergy of patients should be monitored following DSA. In an allergy, prompt and proactive treatment is essential to prevent worsening and dissemination.

Fast-Specific Fluorescent Probes to Visualize Norepinephrine Signaling Pathways and Its Flux in the Epileptic Mice Brain.

Norepinephrine (NE) is synthesized in the locus coeruleus and widely projected throughout the brain and spinal cord. It regulates various actions and consciousness linked to a variety of neurological diseases. A "hunting-shooting" strategy was proposed in this work to improve the specificity and response rate of an NE fluorescent probe: 2-(cyclohex-2-en-1-ylidene)malononitrile derivatives were chosen as a fluorophore. To create a dual-site probe, an aldehyde group was added to the ortho of the ester group (or benzene sulfonate). Because of its excellent electrophilic activity, the aldehyde group could rapidly "hunt" the amino group and then form an intramolecular five-membered ring via the nucleophilic reaction with the ß-hydroxyl group. The -NH- in the five-membered ring "shoots" the adjacent ester group, releasing the fluorophore and allowing for rapid and specific NE detection. The NE release and reuptake ″emetic″-″swallow″ transient process is captured and visualized under the action of the primary NE receptor drug. Furthermore, by introducing halogen into the fluorophore to lengthen the absorption wavelength, improve lipid solubility, and adjust the pKa appropriately, the probe successfully penetrated the blood-brain barrier (BBB). In situ synchronous probe imaging was used to detect the NE level in the brains of epileptic and normal mice, and abnormal expression of NE in the brain was discovered during epilepsy. Brain anatomy was used to examine the distribution and level changes of NE in various brain regions before and after epilepsy. This research provides useful tools and a theoretical foundation for diagnosing and treating central nervous system diseases early.

A nucleophilic addition-elimination based ratiometric fluorescent probe for monitoring N2H4 in biological systems and actual samples.

Hydrazine (N2H4) is an important reagent in the field of fine chemical engineering. However, its accumulation in the environment and food chain could pose a great threat to food safety and human health. Therefore, designing a fluorescent probe with good cell penetration and high selectivity and sensitivity to detect N2H4 in actual samples and in vivo is a meaningful project. Herein, due to the nucleophilicity of hydrazine, we utilized naphthalimide as the fluorescence chromophore and pyrone as the recognition site to achieve the ratiometric detection of hydrazine by ring opening. In addition, we introduced the ester to improve the lipid solubility of the probe, which allowed the probe to better penetrate the cell membrane to realize the fluorescent imaging of probes in cells. Meanwhile, to our delight, the probe showed high selectivity and sensitivity to N2H4 in the test system, so we further applied the probe in water samples and food, in vitro and in vivo.

An ultra-fast UV-electrochemical sensor based on Cu-MOF for highly sensitive and selective detection of ferric ions.

A 2D Cu-MOF: {[CuL(H2O)]}n (Cu-1, H2L = 3,4-ethylene dioxythiophene-2,5-dicarboxylic acid) was synthesized using the hydrothermal method. Cu-1 showed excellent solvent stability and was used to fabricate a UV ferric ion sensor. An ultra-low limit of detection (LOD) at 14.5 fM was obtained. Furthermore, N,N-dimethylformamide (DMF) as a 'turn-off' switch was introduced into the Cu-1 framework to construct another 2D Cu-MOF: {[CuL(DMF)]}n (Cu-2) by a single crystal to single crystal (SCSC) transformation method. Cu-2 lost the ability to recognize ferric ions and the switching effect of Fe3+ recognition was realized. Cyclic voltammograms (CVs) were employed to investigate this conversion process and provided a way for explaining the interaction mechanism between Cu-1 and ferric ions. We present an approach for designing and synthesizing MOFs that are suitable for ion sensing.

A near-infrared fluorescent probe for in situ imaging of SO2 flux in drug-induced liver injury.

Sulfur dioxide (SO2) has been widely applied as an important additive in various foods and drugs due to its antioxidant, antiseptic and bleaching properties. SO2 in living organisms plays a key biological role as an antioxidant in a variety of life activities. However, abnormal levels of SO2 in both food and living organisms could cause harm and even serious illness, such as diseases related to the respiratory and cardiovascular systems and cancers. Therefore, it is of great practical significance to accurately determine the level of SO2 in food and organisms. In this work, we synthesized a novel near-infrared ratiometric fluorescent probe (NTO) using xanthene and benzopyran as the matrix for the detection of SO2. NTO demonstrates a rapid response (within 8 s), high selectivity, excellent sensitivity (LOD = 3.64 µM) and a long emission wavelength (800 nm), which could be applied to SO2 monitoring in a complex environment. NTO showed a high recovery (90%-110%) of SO2 in food samples such as beer and rock sugar. The results of HeLa cell experiments indicate that NTO has excellent fluorescence labeling ability for SO2 in endoexogenous-sulfide metabolism. In addition, we applied it to mice with acetaminophen (APAP)-induced acute liver injury and observed changes in SO2 during liver injury. Based on these results, we believe that this will provide a convenient visual tool for the detection of the SO2 content in food safety and biomedicine.

Eclampsia with hypothyroidism complicated with posterior reversible encephalopathy syndrome-a case report.

BACKGROUND: Posterior reversible encephalopathy syndrome (PRES) is a rare neurological disorder with complex physiopathological mechanisms that have not been fully understood. Early identification is of great prognostic significance, of which the symptoms and radiological abnormalities can be completely reversed. If the diagnosis and treatment are delayed, ischemia and massive infarction may be developed in some patients. Posterior reversible encephalopathy syndrome (PRES) has been reported mainly in association with postpartum eclampsia, which have been rarely reported, while the association with hypothyroidism has not been reported at home or abroad. CASE PRESENTATION: Here we report on a pregnant 29-year-old with multipara and a chief complication of hypothyroidism. She presented in the emergency department with frequent attacks of severe headache symptoms resulting from reversible cerebral vasoconstriction syndrome (RCVS), accompanied with prenatal eclampsia. PRES was determined by radiological examination. CONCLUSION: To the best of our knowledge, this is the first case of PRES complicated by hypothyroidism and prepartum eclampsia.Clinicians should be alert for the co-occurence of eclampsia, PRES, and RCVS when patients have convulsions after a typical throbbing headache. Moreover, regular monitoring of thyroid function during pregnancy should also occupy certain special attention.

Relationships between blood bone metabolic biomarkers and anemia in patients with chronic kidney disease.

INTRODUCTION: Blood bone metabolic biomarkers are noninvasive indices for evaluating metabolic bone diseases. We investigated the relationships between blood bone metabolic biomarkers and anemia in chronic kidney disease (CKD) patients and analyzed the effects of parathyroidectomy (PTX) on the above indices. METHODS: In this cross-sectional study, 100 healthy controls and 239 CKD patients, including 46 secondary hyperparathyroidism (SHPT) patients with PTX, were enrolled. Moreover, a prospective study was conducted in which 28 PTX patients were followed up. The degree of anemia was classified as mild, moderate, or severe based on the tertiles of hemoglobin (Hb) levels of the anemic CKD patients, with cutoff values of 83 g/L and 102 g/L. Bone metabolic biomarkers, including calcium (Ca), phosphorus (P), intact parathyroid hormone (iPTH), fibroblast growth factor 23 (FGF23), and α-klotho, were tested. RESULTS: The mean estimated glomerular filtration rate (eGFR) in CKD patients was 25.7 ± 36.0 ml/min/1.73 m2, and 84.10% of CKD patients had anemia. The baseline Hb levels in the mild, moderate, and severe anemia subgroups were 110.86 ± 5.99 g/L, 92.71 ± 5.96 g/L, and 67.38 ± 10.56 g/L, respectively. CKD patients had higher adjusted Ca, P, alkaline phosphatase (ALP), iPTH, and FGF23 levels and lower α-klotho levels than controls. Baseline adjusted Ca, P, iPTH, and α-klotho levels were associated with Hb levels in CKD patients. Blood adjusted Ca, P, and iPTH levels were correlated with anemia severity. After PTX (median interval: 6.88 months), anemia and high blood adjusted Ca, P, iPTH, and FGF23 levels were ameliorated, while α-klotho levels were increased. CONCLUSIONS: Blood adjusted Ca, P, iPTH, and α-klotho levels were correlated with Hb levels in CKD patients. Correction of bone metabolic disorders may be a therapeutic strategy for anemia treatment.

An Activatable NIR Fluorescent Probe for NAD(P)H and Its Application to the Real-Time Monitoring of p53 Abnormalities In Vivo.

NAD(P)H is crucial for biosynthetic reactions and antioxidant functions. However, the current probes developed for detecting NAD(P)H in vivo require intratumoral injection, which limited their application for animal imaging. To address this issue, we have developed a liposoluble cationic probe, KC8, which exhibits excellent tumor-targeting ability and near-infrared (NIR) fluorescence after reaction with NAD(P)H. By using KC8, it was demonstrated for the first time that the level of NAD(P)H in the mitochondria of living colorectal cancer (CRC) cells was highly related to the abnormality of the p53. Furthermore, KC8 was successfully used to differentiate not only between tumor and normal tissue but also between tumors with p53 abnormality and normal tumors when administered intravenously. Finally, we evaluated tumor heterogeneity through two fluorescent channels after treating a tumor with 5-Fu. This study provides a new tool for real-time monitoring of the p53 abnormality of CRC cells.

Modular Synthesis of a Tridecasaccharide Motif of Bacteroides vulgatus Lipopolysaccharides against Inflammatory Bowel Diseases through an Orthogonal One-Pot Glycosylation Strategy.

Lipopolysaccharides from Bacteroides vulgatus represent interesting targets for the treatment of inflammatory bowel diseases. However, efficient access to long, branched and complex lipopolysaccharides remains challenging. Herein, we report the modular synthesis of a tridecasaccharide from Bacteroides vulgates through an orthogonal one-pot glycosylation strategy based on glycosyl ortho-(1-phenylvinyl)benzoates, which avoids the issues of thioglycoside-based one-pot synthesis. Our approach also features: 1) 5,7-O-di-tert-butylsilylene-directed glycosylation for stereoselective construction of the α-Kdo linkage; 2) hydrogen-bond-mediated aglycone delivery for the stereoselective formation of ß-mannosidic bonds; 3) remote anchimeric assistance for stereoselective assembly of the α-fucosyl linkage; 4) several orthogonal one-pot synthetic steps and strategic use of orthogonal protecting groups to streamline oligosaccharide assembly; 5) convergent [1+6+6] one-pot synthesis of the target.

A Fluorescent Probe for Investigating the Role of Biothiols in Signaling Pathways Associated with Cerebral Ischemia-Reperfusion Injury.

Cerebral ischemia-reperfusion injury (CIRI) is intimately associated with the redox regulation of biothiol, a crucial antioxidant marker that precludes the onset of ROS. We designed a novel fluorescent probe, DCI-Ac-Py, showing various physicochemical properties, such as high selectivity, exceptional signal-to-noise ratio, near-infrared (NIR) optical window, and blood-brain barrier (BBB) penetrability, for detecting biothiols in the brain. The picolinate serves as a specific recognition group that is rapidly activated by biothiol and undergoes nucleophilic substitution with the adjacent acrylic ester to yield the desired NIR probe. Additionally, the probe's lipid solubility is improved through the inclusion of halogen atoms, which aids in penetrating the BBB. Using DCI-Ac-Py, we investigated changes of biothiols in vivo in the brains of mice during CIRI. We found that biothiol-mediated NF-kB classical (P65-related) and nonclassical (RelB-related) pathways contribute to abundant ROS production induced by CIRI and that biothiols are involved in redox regulation. These findings provide new insights into the study of CIRI and shed light on the physiological and pathological mechanisms of biothiols in the brain.

Mitochondrial-Targeting Near-Infrared Fluorescent Probe for Visualizing Viscosity in Drug-Induced Cells and a Fatty Liver Mouse Model.

Mitochondria, as "cell energy stations", are involved in the regulation of various cell functions. Recent investigations revealed that mitochondrial dysfunction that can cause an intracellular viscosity mutation, a process that is associated with an increasing number of diseases that are not curable or manageable. However, conventional viscometers cannot be used to monitor the viscosity changes in living cells and in vivo. In order to cater to the complex biological environment, we present a chemical toolbox, MI-BP-CC, that employs N,N-diethyl and double bonds as sensitive sites for viscosity based on the TICT mechanism (twisted intramolecular charge transfer) to monitor the viscosity of living cells and fatter liver mice. MI-BP-CC features good mitochondrial targeting and a near-infrared emission. Surprisingly, in the presence of viscosity, the MI-BP-CC probe exhibited an ultrasensitive model for viscosity detection showing a red fluorescence signal from a silent "off" state to "on". More importantly, utilizing the satisfactory detection performance of MI-BP-CC, we have successfully visualized increased viscosity under the pathological models of Parkinson's (PD) and fatty liver mice. We anticipate that these findings will provide a convenient and efficient tool to understand physiological functions of viscosity in more biosystems.

Synergistic Modulation by Halogens and Pyridine Crossing the Blood-Brain Barrier for In Situ Visualization of Thiol Flux in the Epileptic Brain.

Epilepsy is a nervous system disease, and seizures are closely related to oxidative stress. Thiols, as the main antioxidant in an organism, play a key role in regulating the redox balance and defending from oxidative stress. As a result of the complexity of the brain structure, there is still a lack of suitable in situ detection methods of thiols to reveal the relationship between epilepsy and thiol level fluctuations. Therefore, by combining picolinate as the new recognition site for thiols, parallel synthesis, and the fluorescence rapid screening method, DCI-Br-3 was developed as a rapid, highly sensitive, and selective probe to monitor thiols in vitro and in vivo. It is worth noting that DCI-Br-3 effectively crossed the blood-brain barrier (BBB) to reveal the negative relationship between the level of thiols and the occurrence of epilepsy and may further provide important information for the prevention and treatment of thiol-related neurological diseases.

Sulfur and nitrogen served as electron donors to rescue short wave emission or fluorescence quenching caused by nucleophilic addition.

SO2 is not only a new potential gas signaling molecule, but also an antioxidant and antioxidant metabolite, which plays an important role in biophysiological and pathological processes. However, excessive intake of it can lead to serious complications of various diseases. Indole-iodide conjugated double bonds are often used as a sensitive response site for SO2, but the fluorescence of the system is often quenched after the nucleophilic addition of SO2. In this work, we introduced sulfur and nitrogen electron donors into a fluorophore, which rescued the trend of fluorescence quenching and enabled the system to develop a ratiometric fluorescence response for more accurate detection of SO32-. In addition, this probe can target mitochondria. The probe has good selectivity, sensitivity, a fast response (200 s) and a limit of detection (LOD) of 0.17 µM. Furthermore, the probe was successfully applied to visualize both endogenous and exogenous SO32- in Hela cells and zebrafish.

Dual-channel detection of viscosity and pH with a near-infrared fluorescent probe for cancer visualization.

Compared to ordinary cells, tumor cells have a unique microenvironment, characterized by high viscosity, low pH, high reactive oxygen species level and the overexpression of certain proteases. Therefore, viscosity and pH can be used as important parameters for visualizing cancer. We designed a spiro-oxazolidine compound (In-1) for the dual-channel detection of viscosity and pH, with the red channel for detecting viscosity and the blue channel for pH. Interestingly, In-1 can locate different organelles under different conditions. Under physiological conditions, In-1 efficiently targeted lysosomes and showed that the viscosity of lysosomes increases in cancer cells while the pH decreases, which can be used to distinguish and detect cancer cells and normal cells. When we treated HL-7702 cells with CCCP, the probe could effectively target the mitochondria, and the fluorescence intensity in the pH channel decreased. This indicates that In-1 can be used as a powerful tool to simultaneously monitor viscosity and pH in different organelles, and may have a guiding role in diseases caused by mitochondrial and lysosomal microenvironments.

A tracer-type fluorescent probe for imaging adenosine triphosphate under the stresses of hydrogen sulfide and hydrogen peroxide in living cells.

Adenosine triphosphate (ATP) is a direct energy source in cells and the core of the biochemical system, and is closely related to various metabolic activities in living organisms. Therefore, designing a simple and rapid ATP detection method is significant to study its physiological function. Herein, a dual-channel fluorescent probe RhB-NA for the in situ imaging of ATP in living cells was designed and synthesized. When ATP bound to RhB-NA, the spirolactam in rhodamine B was induced to open, resulting in a new fluorescence response at 589 nm. Notably, in cell imaging, the treatment of HeLa cells with exogenous H2O2 and H2S, which have certain effects on the mitochondria, confirmed that RhB-NA could detect fluctuations in ATP levels after the mitochondrial state was affected. We believe that RhB-NA has far-reaching significance for studying certain physiological diseases caused by abnormal ATP levels.

Rapid Reaction, Slow Dissociation Aggregation, and Synergetic Multicolor Emission for Imaging the Restriction and Regulation of Biosynthesis of Cys and GSH.

Biosynthesis is a necessary process to maintain life. In recent years, research has fully shown that three kinds of biothiols (Cys, Hcy, GSH) mainly play the role in oxidative stress and maintaining cell homeostasis in cells, and that abnormal concentrations will lead to the occurrence of cardiovascular diseases, cancers, etc. Various fluorescent probes have shown unprecedented advantages in detecting their concentrations and studying their biological functions. As a matter of fact, these three kinds of biothiols are generated in the process of biosynthesis in vivo. It is of great significance to understand their biosynthetic pathways and elucidate their synthetic relationships. In this work, to α,ß-unsaturated ketones conjugated ethylenediamine coumarin and pyrandione was introduced boron fluoride and, through its strong electron deficiency effect, afforded a molecule having near-infrared emission and regulated the rigidity of molecules. At the same time, the conjugated double bond is used to respond to molecular rigidity. The rapid response of the probe to biothiols and the slow dissociation aggregation of the probe itself through the response environment could monitor the absence of biothiols in cells. In addition, based on the difference in sensitivity of response of Cys and GSH to the probe, this work studied the interaction between biosynthetic pathways of Cys and GSH in cells through enzyme inhibition for the first time. The relationship of restriction and regulation of biosynthesis in vivo was revealed.

Molecular-Dimension-Dependent ESIPT Break for Specific Reversible Response to GSH and Its Real-Time Bioimaging.

Glutathione (GSH) plays many important roles in maintaining intracellular redox homeostasis, and determining its real-time levels in the biological system is essential for the diagnosis, treatment, and pathological research of related diseases. Fluorescence imaging has been regarded as a powerful tool for tracking biomarkers in vivo, for which specificity, reversibility, and fast response are the main issues to ensure the real-time effective detection of analytes. The determination of GSH is often interfered with by other active sulfur species. However, in addition to the common features of nucleophilic addition, GSH is unique in its large molecular scale. 2-(2-Hydroxyphenyl) benzothiazole (HBT) was often formed in the ESIPT process. In this study, HBT was installed with α,ß-unsaturated ketone conjugated coumarin derivates or nitrobenzene, which were used to adjust the reactivity of α,ß-unsaturated ketone. Experimental and theoretical calculations found ESIPT to be favorable in HBT-COU but not HBT-COU-NEt2 or HBT-BEN-NO2 due to the higher electronic energies in the keto form. Thus, for HBT-COU, in the presence of GSH, the hydrogen-bonding interaction between C═N of the HBT unit and carboxyl of GSH would inhibit the process, simultaneously promoting the Michel addition reaction between α,ß-unsaturated ketone and GSH. As a consequence, probe HBT-COU could exhibit a rapid reversible ratiometric response to GSH. Small structures of Hcy and Cys are passivated for such reactions. Cell imaging demonstrated the specific response of the probe to GSH, and the probe was successfully used to monitor fluctuations in GSH concentration during cells apoptosis in real-time.

Recent progress of organic small molecule-based fluorescent probes for intracellular pH sensing.

Fluorescent probes along with fluorescence microscopy are essential tools for biomedical research. Various cellular ubiquitous chemical factors such as pH, H2O2, and Ca2+ are labeled and traced using specific fluorescent probes, therefore helping us to explore their physiological function and pathological change. Among them, intracellular pH value is an important factor that governs biological processes, generally ∼7.2. Furthermore, specific organelles within cells possess unique acid-base homeostasis, involving the acidic lysosomes, alkalescent mitochondria, and neutral endoplasmic reticulum and Golgi apparatus, which undergo various physiological processes such as intracellular digestion, ATP production, and protein folding and processing. In this review, recently reported fluorescent probes targeted toward the lysosomes, mitochondria, endoplasmic reticulum, Golgi apparatus, and cytoplasm for sensing pH change are discussed, which involves molecular structures, fluorescence behavior, and biological applications.

Comparing the abundance of HClO in cancer/normal cells and visualizing in vivo using a mitochondria-targeted ultra-fast fluorescent probe.

Organisms are operating and evolving with a highly sophisticated and intelligent defense mechanism to resist bacterial and viral infections. This process involves a variety of reactive oxygen species (ROS), and they coordinate with each other to support different physiological activities. Due to its strong oxidizing properties, hypochlorous acid (HClO), a part of ROS, is a powerful antimicrobial agent in living organisms and exerts a crucial role in the immune system. However, the excessive production of HClO can cause cell damage and even cell death. Herein, we combined benzene-conjugated benzopyrylium as the fluorophore and dimethylthiocarbamoyl chloride as the recognition site to rationally design a probe (BBD). The fluorescence of the probe was quenched based on an effective PET molecular mechanism. Surprisingly, BBD exhibited a turn-on red fluorescence signal for HClO with ultra-fast response (5 s) and high selectivity. Moreover, BBD located mitochondria well and it was found that the abundance of HClO is higher in HeLa cells compared to that in normal cells. Finally, BBD was successfully applied to the visualization of HClO in zebrafish and nude mice.