A specific dual-locked fluorescence probe to visualize the dynamic changes of lipid droplets and hypochlorous acid in inflammation.

Inflammation is a key factor leading to the occurrence and development of many diseases, both lipid droplets (LDs) and hypochlorous acid (HClO/ClO-) are regarded as the important biomarkers of inflammation. Therefore, it is of great significance to develop an efficient single chemical sensor that can simultaneously detect these two biomarkers. To achieve the goal, we developed a dual-locked fluorescence probe (TPA-DNP) by fusing two targets activated reporting system, its implementation was achieved by turning-on the fluorescence of TPA-DNP through LDs and HClO/ClO- simultaneously. In simulated LDs environment, TPA-DNP displayed excellent selectivity to HClO/ClO-, high sensitivity (LOD = 0.527 µM) and strong anti-interference ability. In addition, cell and zebrafish imaging experiments showed that TPA-DNP could be utilized to visualize exogenous/endogenous HClO/ClO- in LDs environment, and could also be used to observe the impact of LDs changes on the HClO/ClO- detection. On the basis, TPA-DNP served as a favorable tool to achieve visualization of inflammatory dynamic changes.

Full-length transcriptomics study of Ustiloxins-induced hepatocyte injury.

Ustiloxins is a mycotoxin produced by the metabolism of Rice false smut. Studies have shown that Ustiloxins may be toxic to animals, but there is still a lack of toxicological evidence. The liver, as the main organ for the biotransformation of foreign chemicals, may be the direct target organ of Ustiloxins toxicity. In this study, we found that cell viability decreased in a dose- and time-dependent manner when BNL CL.2 cells were treated with different concentrations of Ustiloxins (0, 5, 10, 20, 30, 40, 60, 80, 100, 150 and 200 µg/mL) for 24 and 48 h. In addition, scanning electron microscope observation showed that the cell membrane of the experimental group was damaged, with the appearance of apoptotic bodies. Moreover, the ROS and GSH levels were significantly increased in cells exposed to Ustiloxins. We analyzed the key action targets of Ustiloxins on hepatocyte injury using full-length transcriptomics. A total of 1099 differentially expressed genes were screened, of which 473 genes were up-regulated, and 626 genes were down-regulated. Besides, we also found that the expression of MCM7 and CDC45 in BNL CL.2 cells treated with Ustiloxins decreased, and the expression of CCl-2, CYP1b1, CYP4f13, and GSTM1 increased according to qRT-PCR. Ustiloxins might change CYP450 and GST-related genes, affect DNA replication and cell cycle, and lead to oxidative stress and liver cell injury.

Ultrasensitive Electrochemical Platform for Dopamine Detection Based on CoNi-MOF@ERGO Composite.

An electrochemical sensor applied for dopamine (DA) detection was constructed. An easy static way was used to synthesize bimetallic CoNi-MOF. Next, it was mixed with graphene oxide (GO) under ultrasound to get a uniform suspension. Subsequently, the solution was coated on the glassy carbon electrode (GCE) to form CoNi-MOF@ERGO/GCE by the electrochemical reduction method. The interaction between CoNi-MOF and electrochemically reduced graphene oxide (ERGO) enhances the electrocatalytic performance for DA detection. CoNi-MOF@ERGO/GCE has a wider linear range (0.1-400 µM) and a lower detection limit (0.086 µM) under optimum conditions. Furthermore, it has been applied to test DA in human serum samples. The results reveal that the DA sensor shows excellent performance, which will provide a novel idea for more sensitive and quicker DA detection.

TRAF3 deficiency in MDCK cells improved sensitivity to the influenza A virus.

Tumor necrosis factor receptor-associated factor 3 (TRAF3), an adaptor protein, has significant and varying effects on immunity depending on cell types. The role of TRAF3 in Madin-Darby Canine Kidney Epithelial (MDCK) cell resistance to influenza A virus (IVA) remains elusive. In the present study, CRISPR-Cas9 gene editing technology was used to construct the TRAF3 knockout MDCK cells (MDCK-TRAF3-/-). Hemagglutination assay, plaque assay, transcriptome, and quantitative real-time PCR were performed after IVA infection. The results showed that after IVA infection, HA titers and virus titers were promoted, interferon I-related pathways were significantly blocked, and transcription of several antiviral-related genes was significantly decreased in MDCK-TRAF3-/- cells. Thus, our study suggests that TRAF3 gene knockout reduced MDCK cell's resistance to IVA, thereby resulting in a promising way for IVA isolation and vaccine manufacturing.

Synthesis of a Red-Emitting Polarity-Sensitive Fluorescent Probe Based on ICT and Visualization for Lipid Droplet Dynamic Processes.

Abnormal lipid droplets (LDs) have been recognized as critical factors in many diseases because they are metabolically active and dynamic organelles. Visualization for LD dynamic processes is fundamental for elucidating the relationship of LDs and related diseases. Herein, a red-emitting polarity-sensitive fluorescent probe (TPA-CYP) based on intramolecular charge transfer (ICT) was proposed, which was constructed by employing triphenylamine (TPA) and 2-(5,5-dimethyl-2-cyclohex-1-ylidene)propanedinitrile (CYP) as electron donor and acceptor moiety, respectively. The spectra results underlined the excellent characteristics of TPA-CYP, such as high polarity sensitivity (Δf = 0.209 to 0.312), strong solvatochromic effect (λem 595-699 nm), and the large Stokes shifts (174 nm). Moreover, TPA-CYP exhibited a specific ability to target LDs and effectively differentiated cancer cells and normal cells. Surprisingly, TPA-CYP had been successfully applied to dynamic tracking of LDs, not only in inflammation induced by lipopolysaccharide (LPS), the process of oxidative stress, but also in live zebrafish. We believe that TPA-CYP could serve as a powerful tool to gain insight into the dynamics of LDs and to understand and diagnose LD-associated diseases.

Development of an mRNA vaccine against a panel of heterologous H1N1 seasonal influenza viruses using a consensus hemagglutinin sequence.

Seasonal influenza, causes hundreds of thousands of deaths annually, posing a severe threat to human health. Currently available influenza vaccines are targeted only at specific strains or conserved epitopes; however, these vaccines are not completely efficacious because influenza viruses can undergo mutation during circulation, leading to antigenic mismatch between recommended strains and circulating strains and elusion from the immune system. Therefore, developing an influenza vaccine that is quick, effective, and broadly protective has become crucial, and the integral part of hemagglutinin (HA) remains an ideal target for vaccine development. This study developed a lipid nanoparticle-encapsulated nucleoside-modified mRNA vaccine (mRNA-LNPs) encoding a consensus full-length HA sequence (H1c) and evaluated its protective efficacy and immunogenicity through in vitro and in vivo assays. Following two intramuscular immunizations (2, 10 µg, or 20 µg) at a 3-week interval in BALB/c mice, H1c-mRNA-LNP vaccine induced strong antibodies as shown in the hemagglutination-inhibition test and protective neutralizing antibodies against numerous heterologous H1N1 influenza viruses as shown in the microneutralization assay. Additionally, both Th1- and Th2-biased cellular immune responses were elicited, with the Th1-biased response being stronger. Two doses of the H1c-mRNA-LNP vaccine could neutralize a panel of heterologous H1N1 influenza viruses and could confer protection in mice. Taken together, these findings suggest that the H1c-mRNA-LNP vaccine encoding a consensus full-length HA is a feasible strategy for developing a cross-protective vaccine against a panel of heterologous H1N1 influenza viruses.

Type XV osteogenesis imperfecta: A novel mutation in the WNT1 gene, c.620G >A (p.R207H), is associated with an inner ear deformity.

The Wnt signaling pathway is vital in encouraging bone growth. WNT1 gene mutations have been identified as the major cause of type XV osteogenesis imperfecta (OI). Described here is a case of complex heterozygous WNT1 c.620G>A (p.R207H) and c.677C >T (p.S226L) OI caused by a novel mutation at locus c.620G >A (p.R207H). The female patient had type XV OI, distinguished by poor bone density, frequent fractures, a small stature, skull softening, lack of dentine hypoplasia, a brain malformation, and obvious blue sclera. A CT scan of the temporal bone revealed abnormalities of the inner ear, necessitating a hearing aid 8 months after birth. There was no family history of such disorders in the proband's parents. The proband inherited complex heterozygous WNT1 gene variants c.677C>T (p.S226L) and c.620G>A (p.R207H) from her father and mother, respectively. Presented here is a case of OI with inner ear deformation caused by c.620G>A (p.R207H), which is a novel WNT1 site mutation. This case broadens the genetic spectrum of OI and it provides a rationale for genetic testing of mothers and a medical consultation to estimate the risk of fetal illness.

Automated machine learning-based model for the prediction of delirium in patients after surgery for degenerative spinal disease.

OBJECTIVE: This study used machine learning algorithms to identify critical variables and predict postoperative delirium (POD) in patients with degenerative spinal disease. METHODS: We included 663 patients who underwent surgery for degenerative spinal disease and received general anesthesia. The LASSO method was used to screen essential features associated with POD. Clinical characteristics, preoperative laboratory parameters, and intraoperative variables were reviewed and were used to construct nine machine learning models including a training set and validation set (80% of participants), and were then evaluated in the rest of the study sample (20% of participants). The area under the receiver-operating characteristic curve (AUROC) and Brier scores were used to compare the prediction performances of different models. The eXtreme Gradient Boosting algorithms (XGBOOST) model was used to predict POD. The SHapley Additive exPlanations (SHAP) package was used to interpret the XGBOOST model. Data of 49 patients were prospectively collected for model validation. RESULTS: The XGBOOST model outperformed the other classifier models in the training set (area under the curve [AUC]: 92.8%, 95% confidence interval [CI]: 90.7%-95.0%), validation set (AUC: 87.0%, 95% CI: 80.7%-93.3%). This model also achieved the lowest Brier Score. Twelve vital variables, including age, serum albumin, the admission-to-surgery time interval, C-reactive protein level, hypertension, intraoperative blood loss, intraoperative minimum blood pressure, cardiovascular-cerebrovascular disease, smoking, alcohol consumption, pulmonary disease, and admission-intraoperative maximum blood pressure difference, were selected. The XGBOOST model performed well in the prospective cohort (accuracy: 85.71%). CONCLUSION: A machine learning model and a web predictor for delirium after surgery for the degenerative spinal disease were successfully developed to demonstrate the extent of POD risk during the perioperative period, which could guide appropriate preventive measures for high-risk patients.

A novel electrochemical sensor based on AuPd/UiO-66-NH2/GN composites for sensitive dopamine detection.

Metal-organic frameworks (MOFs) have emerged as interesting nanomaterials owing to their large surface area, high porosity, tunable pore architecture and easy functionalization. However, an inferior electrical conductivity hinders their application in electrochemical sensing. In this paper, gold-palladium alloy/UiO-66-NH2/graphene (AuPd/UiO-66-NH2/GN) composites were synthesized by loading alloys on the surfaces of MOFs and then attaching them to the graphene surface. The addition of metal nanoparticles and graphene enhanced the electron transfer ability of MOFs. Then, composites were used to modify a glassy carbon electrode (GCE) to construct a sensitive dopamine (DA) electrochemical sensor. The developed sensor manifested two linear relationships in lower concentration ranges and in higher concentration ranges with a 0.21 × 10-6 mol L-1 low detection limit (3σ/k) under optimal conditions. The results certified that the constructed sensor had high selectivity, excellent reproducibility and good stability, and had been used successfully for DA detection in actual human serum samples.

The Characteristics and Potential Risks of Neonicotinoid Residues in Soils of Different Types of Land Use in Hangzhou.

Due to the wide existence of neonicotinoid insecticides (neonics) and their potential impact on ecosystems and human health, they have received special attention in recent years. Soil is not only a sink of neonics but also a source of neonics, so it plays a key role in the ubiquity of neonics in the environment. The purpose of this research was to compare neonics residues in soils of different types of land use and estimate their exposure to different populations via ingestion. A total of 130 soil samples from six different types were collected. The concentrations of seven neonics in soil were simultaneous determined using isotope-dilution ultra-performance liquid chromatography-tandem mass spectrometry. The results showed that at least one neonic was analyzed in all samples. The highest average concentration was 3.42 ng/g (clothianidin), followed by 3.39 ng/g (thiamethoxam), 3.06 ng/g (acetamiprid), 2.84 ng/g (imidacloprid), 2.66 ng/g (nitenpyram), 2.43 ng/g (thiacloprid), and 1.89 ng/g (dinotefuran). IMI and ACE were the most commonly found neonics in soil. The neonic levels in different soils varied significantly. The integrated neonic residue in cropland was much higher than that in other types of land. The risk assessment revealed that the average daily dose (ADD) through ingestion contact with soil was acceptable to children and adults. With the increasing evidence that neonics could cause a variety of toxic effects on mammals and humans, ingestion exposure caused by neonics in soil should also receive continuous attention in future studies.

Preparation of Nereid oligopeptide and investigation of the mechanism underlying the induction of apoptosis in human lung cancer H1299 cells.

In the present study, oligopeptides from Nereid (Perinereis aibuhitensis) were prepared via enzymatic hydrolysis, and the mechanism underlying the induction of apoptosis in H1299 cells was investigated. According to the analysis of the inhibition rate on proliferation, alkaline protease demonstrated the best enzymatic efficiency. The optimal conditions for hydrolysis were as follows: 50˚C and pH 10 for 6 h; a material­to­liquid ratio of 1:1 (g/ml); and addition of 400 U/g enzyme. The hydrolysates were purified using ultrafiltration, anion chromatography, gel filtration chromatography, and high­performance liquid chromatography. The Nereid oligopeptide (NOP), with a molecular weight of 841 kDa and an amino acid sequence of glutamine­isoleucine­asparagine­glutamine­histidine­leucine, was obtained. NOP inhibited the proliferation of H1299 cells in a time­ and dose­dependent manner. Morphological changes and apoptosis were also induced by NOP in H1299 cells. The western blot analysis revealed that the B­cell lymphoma 2/Bcl­2 associated X (Bcl­2/Bax) ratio was reduced by 24.7% in the NOP treatment group compared with the control group. The relative expression levels of cleaved caspase­9 (cleaved­CASP9) and cleaved caspase­3 (cleaved­CASP3) in the NOP treatment group were 2.55­ and 1.71­fold higher than those measured in the control group, respectively. These results suggested that NOP exerts antitumor effects by influencing the proliferation and apoptosis of H1299 cells.

TICT-Based Microenvironment-Sensitive Probe with Turn-on Red Emission for Human Serum Albumin Detection and for Targeting Lipid Droplet Imaging.

Fluorescent probes sensitive to microenvironment have always been fascinating due to their tremendous advantages in tracking changes in the pathophysiological microenvironment and potential application in the early diagnosis of related diseases. In this study, a fluorescent luminogen, triphenylamine-thiophene-rhodanine (TPA-TRDN), with high sensitivity to changes in polarity and viscosity was designed and could be applied to detecting human serum albumin (HSA) in actual urine, as well as lipid droplets (LDs) in cells and in vivo with turn-on red emission. TPA-TRDN could selectively detect HSA with fast response (10 min), superior sensitivity (LOD 0.34 µg/mL, about 60-fold fluorescence enhancement), and wide detection range (0.00-0.30 mg/mL). The detection mechanism was demonstrated: TPA-TRDN encountered the hydrophobic IB domain of HSA, leading to the inhibition of the twisted intramolecular charge transfer (TICT) phenomenon and intramolecular rotation. Moreover, TPA-TRDN demonstrated satisfactory ability to identify cancer cells and noncancer cells by microenvironment-guided specific LD bioimaging. This evidence indicated that TPA-TRDN has promising application in the microenvironment-related biomedical field and clinical diagnosis.

Synthesis of a new environment-sensitive fluorescent probe based on TICT and application for detection of human serum albumin and specific lipid droplets imaging.

Environment-sensitive fluorescent probes have always been as forceful tools to understand the pathophysiological processes of relevant diseases. In this work, a new fluorescent probe with typical D-π-A structure was designed and showed high sensitivity to polarity and viscosity changes. DPAR could selectively detect human serum albumin (HSA) with turn-on orange emission in aqueous PBS buffer (pH 7.4), which showed advantages such as rapid response (4 min), high sensitivity (LOD 0.98 µg/mL). Therefore, it was successfully used for achieving HSA levels in urine samples and HSA imaging in HeLa cells. DPAR also exhibited the capability to recognize the cancer cells over the normal cells by lower polarity guided lipid droplets (LDs) imaging (in green emission channel). The detection mechanism for HSA and cancer diagnosis was convinced that DPAR encountered the lower-polarity and higher-viscosity microenvironment, resulting in the confinement of the TICT process and intramolecular rotation. These facts showed that DPAR had good application prospects in environment-related biomedical research and clinical diagnosis.

Lysosome targeting, Cr(vi) and l-AA sensing, and cell imaging based on N-doped blue-fluorescence carbon dots.

N-doped blue-fluorescence carbon dots (N-CDs) were fabricated via a one-pot hydrothermal method using folic acid and p-phenylenediamine. The obtained N-CDs exhibited strong fluorescence (FL) with a considerable quantum yield (QY) of 21.8% and exceptional optical stability under different conditions. Upon introducing Cr(vi), blue FL of N-CDs was distinctly quenched. On subsequent addition of l-AA, the FL of N-CDs could be partially recovered. The fluorescence changes of N-CDs have been utilized to detect Cr(vi) and l-AA in aqueous solutions with linear ranges of 0.10-150 µM and 0.75-2.25 mM, respectively, as well as limit of detection values of 9.4 nM and 25 µM, respectively. Furthermore, as-obtained N-CDs can be extended to monitor the fluctuation of intracellular Cr(vi) and l-AA. More intriguingly, N-CDs can target lysosomes with a satisfactory Pearson correction coefficient of 0.87, which indicates a promising application prospect in the biomedical field.

Development of a piperazinyl-NBD-based fluorescent probe and its dual-channel detection for hydrogen sulfide.

To selectively detect H2S based on the thiolysis reaction of 7-nitro-1,2,3-benzoxadiazole (NBD), amines attracted increasing attention since NBD amine is regarded as a new H2S reaction site. Herein, a novel fluorescent probe, triphenylamine piperazine NBD (TPA-Pz-NBD), was developed. The results showed that it exhibited high selectivity towards H2S via fluorescence spectroscopy and solution color. Furthermore, TPA-Pz-NBD not only detected H2S by a dual-channel, turn-on fluorescence signal at 500 nm and turn-off fluorescence signal at 545 nm, respectively, but also displayed a wide detection range of 0-125 µM. In addition, living cell imaging results indicated that TPA-Pz-NBD holds potential for the detection of intracellular H2S.

A red emitting fluorescent probe based on TICT for selective detection and imaging of HSA.

A red emitting fluorescence probe, TPA-CPO, based on twisted intra-molecular charge transfer (TICT) was designed and synthesized. The spectra results displayed that TPA-CPO could sense HSA with excellent properties including significant fluorescence enhancement, long emission wavelength, large stokes shift, and wide linear range. The recognition mechanism was proved that TPA-CPO could bind to domain IB of HSA and its TICT process was suppressed by utilizing hydrophobic cavity and low polarity of HSA. TPA-CPO bind to domain IB instead of common drug sites of HSA could effectively avoid interference from most drugs. The selective response of TPA-CPO allowed quantitative detection of HSA with sensitivity limit of 13.65 µg/mL. What's more, it successfully achieved HSA imaging in HeLa cells.

Orange emissive carbon nanodots for fluorescent and colorimetric bimodal discrimination of Cu2+ and pH.

We have facilely synthesized orange emissive carbon nanodots (O-CDs) via a hydrothermal method using citric acid and 5-aminosalicylic acid. The obtained O-CDs show the excellent characteristics of excitation independence, low toxicity, fabulous photostability and superior biocompatibility. Based on these captivating properties, as-prepared O-CDs have been successfully implemented as a multi-functional sensing platform for fluorescent and colorimetric bimodal recognition of Cu2+ and pH. Upon adding Cu2+, the orange fluorescence of the O-CDs is evidently quenched with a linear range of 0 µM-300 µM, and a detection limit of 28 nM. Additionally, as the pH increases from 7.0 to 10.2, the O-CDs manifest an obvious decrease in orange fluorescence, which shows a pKa value of 8.73 and excellent linearity in the pH range of 8.0-9.2. Appealingly, the laser confocal imaging of O-CD-stained cells demonstrates that the fluctuations of Cu2+ and pH can be visualized in living cells.

Synergistic Effects of Ternary PdO-CeO2-OMS-2 Catalyst Afford High Catalytic Performance and Stability in the Reduction of NO with CO.

We developed a robust ternary PdO-CeO2-OMS-2 catalyst with excellent catalytic performance in the selective reduction of NO with CO using a strategy based on combining components that synergistically interact leading to an effective abatement of these toxic gases. The catalyst affords 100% selectivity to N2 at the nearly full conversion of NO and CO at 250 °C, high stability in the presence of H2O, and a remarkable SO2 tolerance. To unravel the origin of the excellent catalytic performance, the structural and chemical properties of the PdO-CeO2-OMS-2 nanocomposite were analyzed in the as-prepared and used state of the catalyst, employing a series of pertinent characterization methods and specific catalytic tests. The experimental as well as theoretical results, based on density-functional theory calculations suggest that CO and NO follow different reaction pathways, CO is preferentially adsorbed and oxidized at Pd sites (PdII and Pd0), while NO decomposes on the ceria surface. Lattice oxygen vacancies at the interfacial perimeter of PdO-CeO2 and PdO-OMS-2, and the diffusion of oxygen and oxygen vacancies are proposed to play a critical role in this multicenter reaction system.

A turn-on fluorescence probe for hydrogen sulfide in absolute aqueous solution.

A turn-on hydrogen sulfide (H2S) fluorescence probe, 4-{2-[4-(2,4-dinitrophenoxy)-phenyl]-vinyl}-1-methyl-pyridinium iodide (DPPVP), based on the thiolysis reaction of dinitrophenyl ethers (DNP) has been proposed. Pyridinium structure enhanced the water solubility of DPPVP, which could quickly respond to H2S in absolute PBS solution and the fluorescence spectra of DPPVP at 520 nm were turned on by H2S. The spectra results exhibited that DPPVP could sensitively detect H2S with satisfied linear range (0-40 µM) and detection limit (13.4 nM). The high selectivity for H2S against biothiols was attributed to the significant difference in the pKa and the molecular size. Moreover, DPPVP has been successfully used for detecting H2S in vegetable.

Smilax China-derived yellow-fluorescent carbon dots for temperature sensing, Cu2+ detection and cell imaging.

Here, we report an environmentally friendly fabrication strategy of bright yellow fluorescent carbon dots (y-CDs) and construct a rapid and accurate multifunctional sensing platform for the effective detection of temperature and Cu2+. The y-CDs were favorably obtained through a one-step hydrothermal treatment of natural Smilax China for the first time and exhibit long-wavelength emission at 542 nm under an excitation wavelength of 470 nm. Additionally, the obtained y-CDs exhibit superior biocompatibility and distinguished stability under different conditions, and display a respectable fluorescence quantum yield of up to 22.37%. Appealingly, the as-prepared y-CDs were implemented as temperature probes within ranges of 25 °C-40 °C and 45 °C-80 °C. Significantly, based on the static quenching effect, the as-prepared y-CDs were developed as an effective platform for fluorescence sensing of Cu2+, with linear ranges of 0.5 µM-10 µM, 75 µM-225 µM and 250 µM-350 µM, achieving a detection limit of 28 nM. Furthermore, confocal fluorescence imaging of PC12 cells was achieved successfully, which indicated that the as-synthesized y-CDs could visualize Cu2+ fluctuations in living cells.