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Xanthine dehydrogenase, a member of the molybdenum enzyme family, participates in purine metabolism and catalyzes the generation of ureides from xanthine and hypoxanthine. However, the mechanisms by which xanthine dehydrogenase affects rice growth and development are poorly understood. In the present study, we identified a mutant with early leaf senescence and reduced tillering that we named early senescence and less-tillering 1 (esl1). Map-based cloning revealed that ESL1 encodes a xanthine dehydrogenase, and it was expressed in all tissues. Chlorophyll content was reduced and chloroplast maldevelopment was severe in the esl1 mutant. Mutation of ESL1 led to decreases in allantoin, allantoate, and ABA contents. Further analysis revealed that the accumulation of reactive oxygen species in esl1 resulted in decreased photosynthesis and impaired chloroplast development, along with increased sensitivity to abscisic acid and abiotic stresses. Ttranscriptome analysis showed that the ESL1 mutation altered the expression of genes involved in the photosynthesis process and reactive oxygen species metabolism. Our results suggest that ESL1 is involved in purine metabolism and the induction of leaf senescence. These findings reveal novel molecular mechanisms of ESL1 gene-mediated plant growth and leaf senescence.

本研究采用密度泛函理论（DFT）研究了在γ-Fe2O3表面HCl对Hg0的吸附和催化氧化的作用机制。构建了Hg0、HCl、HgCl和HgCl2在γ-Fe2O3(001)表面的吸附模型，分析了HCl对γ-Fe2O3表面催化氧化Hg0的作用机理，并通过反应路径的能量分布测定，研究了γ-Fe2O3表面Hg0的氧化过程。结果表明，Hg0倾向于化学吸附在γ-Fe2O3(001)表面Feoct位。HCl在催化剂表面进行解离吸附，形成吸附态Cl和羟基，从而促进Hg0的吸附。HgCl以分子形式化学吸附在γ-Fe2O3(001)上，并作为Hg0氧化过程的中间体。HgCl2倾向在γ-Fe2O3表面上的平行吸附。同时，HCl在γ-Fe2O3(001)上氧化Hg0遵循L-H机理，即化学吸附的Hg0与解离吸附的HCl反应，且HCl对Hg0的非均相氧化通过两步反应途径进行，即Hg0 (ads)→HgCl(ads)→HgCl2(ads)。

Let G = ( V , E ) be a simple graph. The inverse sum indeg index of G is defined as I S I ( G ) = ∑ u v ∈ E ( G ) d ( u ) d ( v ) d ( u ) + d ( v ) , where d ( u ) is the degree of u . This index has a nice predicting ability for the total surface area of octane isomers. A cactus is a connected graph in which all of its blocks are either edges or cycles. Let C n , k be the set of cacti of order n and with k cycles. In this note, we characterize all cacti with minimum inverse sum indeg index among C n , k when n ≥ 6 k − 3 .

An efficient tandem nucleophilic aminopalladation and carbene insertion sequence is described for the synthesis of indole fused polycycles. The reaction process provides a variety of substituted indeno[1,2-b]indoles in up to 99% yields.

Palladium-catalyzed C-N bond formation is one of the widely used transformations for the synthesis of structurally diverse N-heterocycles. This work describes an efficient palladium-catalyzed multiple-C-N bond formation reaction for the synthesis of highly π-conjugated N-heterocycles, indolo[3,2-b]indoles with di-tert-butyldiaziridinone. The reaction likely proceeds through the initial formation of an indole-fused palladacycle by nucleophilic aminopalladation and subsequent bisamination to give indolo[3,2-b]indoles.

The effect of high pressure processing (HPP, 600 MPa, 5 min) and thermal treatment (85 °C, 15 min) on aronia berry juice in a pilot scale was studied. The maximal shelf-life of treated samples at room temperature (RT, approximately 25 °C) or refrigerated storage (RS, 4 °C) was also investigated. Microbial counts, physicochemical properties, enzymes activities, phenolic compounds, and antioxidant activities of these juices were determined and compared. Results indicated that HPP treatment improved the microbial shelf-life of the aronia juice by at least 10-times at RT and 5-times at RS. Although thermal treatment was equally effective in extending the shelf-life, the high temperature resulted in a quicker degradation of polyphenols in aronia juice, which was decreased by 36.6% during RT storage (5 weeks) and 43.3% at RS storage (24 weeks). Therefore, HPP was more efficient in maintaining the safety and quality of aronia juice. The study also indicated HPP treated aronia juice could be stored at RT for at least one month that could be of benefit to the non-cold chain process which is targeting for a low-energy input while still retaining a minimal effect on the nutritional properties of food products.

As a new treatment technique, photothermal therapy (PTT) has aroused worldwide attention in cancer treatment, mainly due to its excellent absorption ability, easy regulation, and biodegradability. Photothermal conversion materials with enhanced permeability and retention effect can be targeted easily to tumor tissue. They can accumulate efficiently to tumor tissues and allow normal tissues and organs not to be affected by temperature, thus significantly helping to reduce the systemic toxicity and improve the antitumor effect. However, PTT alone often suffers from therapeutic resistance and reduced therapeutic efficacy, due to photothermal nanomaterial-mediated fundamental cellular defense mechanism of heat shock response, which could be inhibited by small interfering RNA (siRNA). Nevertheless, photothermal conversion materials as an excellent siRNA delivery carrier may considerably enhance the delivery efficiency of siRNA. Therefore, photothermal and RNA interfering (RNAi) synergistic therapy has recently aroused extensive attention in tumor treatment. In this review, we mainly summarize the recent advances of photothermal and RNAi synergistic therapy, including some synergistic therapeutic nanoplatforms of inorganic and organic photothermal materials and other combined therapies such as combining with small molecular antitumor agents or PDT/imaging. The combination of various treatment techniques may considerably improve the synergistic therapeutic effect of PTT and RNAi in the treatment of cancers.

In this work, the titanium germanosilicide (TiSiGex) superlattice (SL) has been successfully fabricated. A monolayer of silicon atoms and bilayer of inversed titanium silicide constructed this novel superlattice periodically. A localized strain field has been found as a crucial factor via high resolution Annular Dark Field Scanning Transmission Electron Microscope (ADF-STEM) images, being generated by gradual segregation of germanium atoms. Germanium atoms would be excluded during the formation of the transition silicide. This phenomenon could be interpreted by thermodynamic preference. There was a substitution reaction between silicon and germanium, resulting from similar atomic volumes of both. In other words, germanium segregation pathway was based on where substitution occurred. Eventually, the excluded germanium atoms tended to accumulate at the boundary of TiSiGex-SL, contributing to a discontinuous thin film layer.

This work discloses a Cu(II)–Ni(II) catalyzed tandem hydrocarboxylation of alkynes with polysilylformate formed from CO2 and polymethylhydrosiloxane that affords α,β-unsaturated carboxylic acids with up to 93% yield. Mechanistic studies indicate that polysilylformate functions as a source of CO and polysilanol. Besides, a catalytic amount of water is found to be critical to the reaction, which hydrolyzes polysilylformate to formic acid that induces the formation of Ni–H active species, thereby initiating the catalytic cycle.

The assembly of DNA with metal-complex cofactors can form promising biocatalysts for asymmetric reactions, although catalytic performance is typically limited by low enantioselectivities and stereo-control remains a challenge. Here, we engineer G-quadruplex-based DNA biocatalysts for an asymmetric cyclopropanation reaction, achieving enantiomeric excess (eetrans) values of up to +91% with controllable stereoinversion, where the enantioselectivity switches to −72% eetrans through modification of the Fe-porphyrin cofactor. Complementary circular dichroism, nuclear magnetic resonance, and fluorescence titration experiments show that the porphyrin ligand of the cofactor participates in the regulation of the catalytic enantioselectivity via a synergetic effect with DNA residues at the active site. These findings underline the important role of cofactor modification in DNA catalysis and thus pave the way for the rational engineering of DNA-based biocatalysts.

Phloretin, a dihydrochalcone, widely exists in the fruits of apple trees and crabapple trees (Malus prunifolia) with multiple biological activities. Presently, we studied the function of phloretin on the attenuation of hepatic steatosis and further explored the underlying mechanisms both in vitro and in vivo. Male C57BL/6J mice were fed a normal diet or high fat diet (HFD) with or without phloretin (100 mg kg-1) for 12 weeks. HepG2 cells were induced by 200 μM palmitic acid (PA) and co-incubated with phloretin (50 μM) for 24 h. The results showed that phloretin treatment significantly decreased the accumulation of lipids in the liver of the HFD-fed C57BL/6J mice and PA-induced HepG2 cells. Also, phloretin effectively ameliorated hepatic steatosis via promoting fatty acid β-oxidation (FAO). This biological activity of phloretin was closely related to its capacity to improve mitochondrial dysfunction, including the promotion of mitochondrial biosynthesis and inhibition of mitochondrial swelling through the AMPK-dependent SIRT1/PGC-1α and SIRT3/CypD signaling pathways, respectively. These results demonstrate that phloretin effectively improves mitochondrial function and ameliorates HFD-induced hepatic steatosis through an AMPK-dependent signaling pathway.

BACKGROUND: DNA methylation balance is an important regulatory mechanism for mammalian and plant development. The fruit ripening and anthocyanin accumulation of Vaccinium corymbosum are complex developmental processes that involve numerous physiological, biochemical, and structural alterations. OBJECTIVE: This study aimed to investigate the correlation of DNA methylation balance, DNA methylation and demethylation-related gene expression models and anthocyanin accumulation during blueberry fruit ripening. METHODS: The anthocyanin contents during V. corymbosum ‘O’Neal’ fruit development were evaluated. The V. corymbosum DNA methylation- and anthocyanin accumulation-related genes were isolated, and their relative expression patterns were detected during flower bud enlargement and fruit development. Moreover, the relative expression patterns of anthocyanin biosynthetic genes and the dynamic changes in the DNA methylation of the promoter sequences of key anthocyanin biosynthetic genes were evaluated. RESULTS: The results showed that the DNA methylation level of V. corymbosum fruit was consistent with anthocyanin accumulation during ripening, and the expression levels of anthocyanin biosynthetic and DNA methylation-related genes. CONCLUSIONS: During V. corymbosum fruit ripening, anthocyanin accumulation is regulated partially by DNA methylation balance of VcCHS and VcANS promoters.

Author(s): Lu, Mei; Bowlus, Christopher L; Lindor, Keith; Rodriguez-Watson, Carla V; Romanelli, Robert J; Haller, Irina V; Anderson, Heather; VanWormer, Jeffrey J; Boscarino, Joseph A; Schmidt, Mark A; Daida, Yihe G; Sahota, Amandeep; Vincent, Jennifer; Li, Jia; Trudeau, Sheri; Rupp, Loralee B; Gordon, Stuart C; FOLD Investigators | Abstract: BackgroundBiopsy remains the gold standard for determining fibrosis stage in patients with primary biliary cholangitis (PBC), but it is unavailable for most patients. We used data from the 11 US health systems in the FibrOtic Liver Disease Consortium to explore a combination of biochemical markers and electronic health record (EHR)-based diagnosis/procedure codes (DPCs) to identify the presence of cirrhosis in PBC patients.MethodsHistological fibrosis staging data were obtained from liver biopsies. Variables considered for the model included demographics (age, gender, race, ethnicity), total bilirubin, alkaline phosphatase, albumin, aspartate aminotransferase (AST) to platelet ratio index (APRI), Fibrosis 4 (FIB4) index, AST to alanine aminotransferase (ALT) ratio, and g100 DPCs associated with cirrhosis/decompensated cirrhosis, categorized into ten clusters. Using least absolute shrinkage and selection operator regression (LASSO), we derived and validated cutoffs for identifying cirrhosis.ResultsAmong 4328 PBC patients, 1350 (32%) had biopsy data; 121 (9%) were staged F4 (cirrhosis). DPC clusters (including codes related to cirrhosis and hepatocellular carcinoma diagnoses/procedures), Hispanic ethnicity, ALP, AST/ALT ratio, and total bilirubin were retained in the final model (AUROC=0.86 and 0.83 on learning and testing data, respectively); this model with two cutoffs divided patients into three categories (no cirrhosis, indeterminate, and cirrhosis) with specificities of 81.8% (for no cirrhosis) and 80.3% (for cirrhosis). A model excluding DPCs retained ALP, AST/ALT ratio, total bilirubin, Hispanic ethnicity, and gender (AUROC=0.81 and 0.78 on learning and testing data, respectively).ConclusionAn algorithm using laboratory results and DPCs can categorize a majority of PBC patients as cirrhotic or noncirrhotic with high accuracy (with a small remaining group of patients' cirrhosis status indeterminate). In the absence of biopsy data, this EHR-based model can be used to identify cirrhosis in cohorts of PBC patients for research and/or clinical follow-up.

The effects of polyphenols on the physicochemical properties and in vitro digestibility of Tartary buckwheat starch (TBS) remain scarce. In this study, the rheological, thermal properties, and in vitro digestibility of pregelatinized TBS (pre-TBS) with quercetin complexation at various concentrations were characterized. It was found that quercetin complexation increased the shearing resistance and viscosity of pre-TBS. Both SEM and TGA results revealed a more compact and stable structure of starch-quercetin complex in comparison to pre-TBS. The non-inclusive complex with higher crystallinity was formed through hydrogen bonds, which showed by XRD and FT-IR results. Additionally, complexes exhibited the lower digestion rate and digestion velocity constant, and the resistant starch content of complex (with 10% quercetin addition) was the highest. Therefore, quercetin complexation could improve the thermal and rheological properties, and decrease in vitro digestibility of pre-TBS, which could provide a theoretical basis for functional food development.

Rice is a key crop, and metal pollution is an important factor affecting rice yield and quality. Excavating the QTLs related to the accumulation of rice grain metal content, and analyzing its candidate genes, and then improving the ability of rice to withstand metal ion stress through genetic breeding can effectively reduce the caused by metal pollution in soil. In this study, a japonica rice variety Nekken was used as the female parent, and an indica rice variety HZ was used as the male parent. After F1 generation was obtained by cross-breeding, successive selfings were performed for 120 recombinant inbred lines (RIL population) for experimental materials. Cultured under the same and suitable conditions, routine management, then we measured the metal ion content in rice grains of each line at the maturity stage. A genetic map was constructed using the RIL population to map the QTLs resistant to metal ion stress in rice. As a result, a total of 9 QTLs related to metal ion content were detected. It contains 1 multi-effect QTL related to Cu and Hg; 3 QTLs related to As; 2 QTLs related to Fe, one of which has a LOD value as high as 5.53; 2 QTLs related to Zn accumulation, The LOD of one of them is as high as 7.29. Quantitative analysis of candidate genes related to resistance to metal ion stress in these QTL intervals found that the expression levels of the four genes LOC_Os01g14440 , LOC_Os01g18584 , LOC_Os01g20160 , LOC_Os04g34600 were significantly different between parents, Combining the results of the accumulated data of parental concentrations of different metal ions, we speculated that the high expression of LOC_Os01g14440 , LOC_Os01g18584 , and LOC_Os01g20160 may greatly improve the absorption capacity and the tolerance of rice to Zn stress, while the high expression of LOC_Os04g34600 may increase the absorption capacity and the tolerance of rice to Fe stress. Through the excavation and analysis of this QTL, it was found that these genes may affect the ability of rice to withstand metal ion stress, creating conditions for further breeding and screening of rice varieties resistant to metal ion stress.

Developing high-performance DNA-based biocatalysts for desired stereoselective syntheses remains a formidable challenge. Here, we report promising DNA-based catalysts comprised of G-quadruplex (G4)...

Biguanides (i.e. metformin, phenformin and buformin) are antidiabetic drugs with potential antitumor effects. Herein, a polycationic polymer, N,N′-bis(cystamine)acrylamide-buformin (CBA-Bu), contai...

Severe anorexia limits the clinical application of cisplatin, and even leads to the discontinuation of treatment. However, the mechanisms underlying cisplatin-induced anorexia are unknown. Herein, we demonstrated that cisplatin could affect neuronal gamma oscillations and induce abnormal neuronal theta-gamma phase-amplitude coupling in the arcuate nucleus (Arc) of the hypothalamus, and these findings were associated with significantly decreased food intake and weight loss in mice. Chemogenetic activation of AgRP neurons in the Arc reversed the cisplatin-induced food intake reduction in mice. We further demonstrated that endothelial peroxynitrite (ONOO−) formation in the Arc induced nitrosative stress following cisplatin treatment via a previously uncharacterized pathway involving neuronal caspase-1 activation. Strikingly, treatment with the ONOO− scavenger uric acid (UA) reversed the reduced action potential (AP) frequency of AgRP neurons and increased the AP frequency of POMC neurons induced by SIN1, a donor of ONOO−, in the Arc, as determined by whole-cell patch-clamp electrophysiological recording. Consistent with these findings, UA treatment effectively alleviated cisplatin-induced dysfunction of neuronal oscillations and neuronal theta-gamma phase-amplitude coupling in the Arc of mice. Taken together, these results suggest, for the first time, that targeting the overproduction of endothelial ONOO− can regulate cisplatin-induced neurotoxicity through neuronal caspase-1, and thereby serve as a potential therapeutic approach to alleviate chemotherapy-induced anorexia and weight loss., Highlights • Endothelial ONOO– induced the abnormal neuronal oscillations following cisplatin treatment through caspase-1 in the Arc. • ONOO– scavenger UA could attenuate cisplatin-induced neurotoxicity and caspase-1 activation in the Arc. • Targeting endothelial ONOO– provided a promising approach to alleviate chemotherapy-induced anorexia and weight loss.

Summary p-Methyl benzaldehyde (p-MBA) is a class of key chemical intermediates of pharmaceuticals. Conventional industrial processes for p-MBA production involve the consecutive photochlorination, amination, and acid hydrolysis of petroleum-derived p-xylene, while producing vast pollutants and waste water. Herein, we report a direct, green route for selective synthesis of p-MBA from acetaldehyde using a diphenyl prolinol trimethylsilyl ether catalyst. The optimized p-MBA selectivity is up to 90% at an acetaldehyde conversion as high as 99.8%. Intermediate structure and 18O-isotope data revealed that the conversion of acetaldehyde to p-methylcyclohexadienal intermediates proceeds in an enamine-iminium intermediate mechanism. Then, controlled experiments and D-isotope results indicated that the dehydrogenation of p-methylcyclohexadienal to p-MBA and H2 is catalyzed by the same amines through an iminium intermediate. This is an example that metal-free amines catalyze the dehydrogenation (releasing H2), rather than using metals or stoichiometric oxidants., Graphical abstract, Highlights • A direct route to produce p-methyl benzaldehyde from biomass-derived acetaldehyde • Revealing the reaction kinetics and mechanism under reaction conditions • An example of an organic amine-catalyzed dehydrogenation-aromatization reaction

The influence of strontium doping on the adsorption of methanol on calcium oxide surface was investigated by molecular simulation. The model for methanol adsorption onto the CaO(100) and CaO(100)-Sr surfaces was constructed; the adsorption energy and activation energy were then calculated and the density of states was portrayed for the methanol bond on the calcium oxide surface. The methanol activation degrees on the calcium oxide surface before and after strontium doping were then compared by analyzing the Mulliken atomic charge population and deformation density. The results illustrate that the adsorption of methanol onto the calcium oxide surface can be significantly enhanced through the strontium doping; moreover, the enhancement increases with an increase in the doping content of strontium. After doping calcium oxide with strontium, the energy required for methanol activation is reduced; as a result, the strontium doping can also enhance the activation degree of methanol, as methanol is activated upon adsorption onto the calcium oxide surface.

CRISPR/Cas-based mRNA imaging has been developed to labeling of high-abundance mRNAs. A lack of non-genetically encoded mRNA-tagged imaging tools has limited our ability to explore the functional distributions of endogenous low-abundance mRNAs in cells. Here, we developed a CRISPR-Sunspot method based on the SunTag signal amplification system that allows efficient imaging of low-abundance mRNAs with CRISPR/Cas9. Methods: We created a stable TRE3G-dCas9-EGFP cell line and generated an Inducible dCas9-EGFP imaging system for assessment of two factors, sgRNA and dCas9, which influence imaging quality. Based on SunTag system, we established a CRISPR-Sunspot imaging system for amplifying signals from single-molecule mRNA in live cells. CRISPR-Sunspot was used to track co-localization of Camk2a mRNA with regulatory protein Xlr3b in neurons. CRISPR-Sunspot combined with CRISPRa was used to determine elevated mRNA molecules. Results: Our results showed that manipulating the expression of fluorescent proteins and sgRNA increased the efficiency of RNA imaging in cells. CRISPR-Sunspot could target endogenous mRNAs in the cytoplasm and amplified signals from single-molecule mRNA. Furthermore, CRISPR-Sunspot was also applied to visualize mRNA distributions with its regulating proteins in neurons. CRISPR-Sunspot detected the co-localization of Camk2a mRNA with overexpressed Xlr3b proteins in the neuronal dendrites. Moreover, we also manipulated CRISPR-Sunspot to detect transcriptional activation of target gene such as HBG1 in live cells. Conclusion: Our findings suggest that CRISPR-Sunspot is a novel applicable imaging tool for visualizing the distributions of low-abundance mRNAs in cells. This study provides a novel strategy to unravel the molecular mechanisms of diseases caused by aberrant mRNA molecules.

Insulin resistance in the brain is a pathological mechanism that is shared between Alzheimer's disease (AD) and type 2 diabetes mellitus (T2DM). Although aberrant expression and phosphorylation of insulin receptor substrate 1 (IRS-1) contribute to insulin resistance, the underlying mechanism remains elusive. In this study, we used several approaches, including adeno-associated virus-based protein overexpression, immunoblotting, immunoprecipitation, immunohistochemistry, and in situ proximal ligation assays, to investigate the function of dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) in IRS-1 regulation and the downstream insulin signaling in neurons. We found that DYRK1A overexpression up-regulated IRS-1 expression by slowing turnover of the IRS-1 protein. We further observed that DYRK1A directly interacted with IRS-1 and phosphorylated IRS-1's multiple serine residues. Of note, DYRK1A and IRS-1 were coordinately up-regulated in the prefrontal cortex of db/db mice brain. Furthermore, DYRK1A overexpression ameliorated chronic high insulin-induced insulin resistance in SH-SY5Y cells as well as in primary rat neurons. These findings suggest that DYRK1A protects against insulin resistance in the brain by elevating IRS-1 expression.

We have designed and synthesized a novel cationic polymer, poly[(2-acryloyl)ethyl (p-boronic acid)benzyl diethylammonium bromide], whose p-boronic acid benzyl can be converted to anionic poly(acrylic acid). Herein, the complexation of the cationic polymer with DNA and their disassembly triggered by reactive oxygen species were analyzed in real time by a high-resolution surface plasmon resonance. We have found that both single-stranded DNA and double-stranded DNA could bind to cationic polymer and immobilized onto the Au surface via electrostatic interactions with the binding constant 1.4 × 1011 M–1 for double-stranded DNA and 1.0 × 1011 M–1 for single-stranded DNA. Upon adding H2O2, the adsorbed DNA was quickly released due to cationic polymer turned into negatively charged poly(acrylic acid). Thus, this study confirmed that the polymer that can undergo positive to negative charge-reversal and provide the quick release of DNA for effective gene transfection.

The study evaluated effects of cholesterol supplementation in a diet with high soybean meal (SBM) on the growth and cholesterol metabolism of giant grouper (Epinephelus lanceolatus). All‐fish‐meal diet was used as control. The diet including SBM (replaced 50% of the fish meal protein, SBM diet) and the SBM diet supplemented with 10 g/kg cholesterol (SBM + cholesterol) were used as experimental diets. Three diets were each fed to triplicate groups of juvenile grouper (initial body weight: 12.39 ± 0.36 g) in a recirculating aquaculture system for 8 weeks. Grouper fed the control diet showed higher (p

Exosomes are naturally secreted nanovesicles that have emerged as a promising therapeutic nanodelivery platform, due to their specific composition and biological properties. However, challenges like considerable complexity, low isolation yield, drug payload, and potential safety concerns substantially reduce their pharmaceutical acceptability. Given that the nano-bio-interface is a crucial factor for nanocarrier behavior and function, modification of synthetic nanoparticles with the intrinsic hallmarks of exosomes' membrane to create exosome mimetics could allow for siRNA delivery in a safer and more efficient manner. Herein, connexin 43 (Cx43)-embedded, exosome-mimicking lipid bilayers coated chitosan nanoparticles (Cx43/L/CS NPs) were constructed by using cell-free (CF) synthesis systems with plasmids encoding Cx43 in the presence of lipid-coated CS NPs (L/CS NPs). The integration of de novo synthesized Cx43 into the lipid bilayers of L/CS NPs occurred cotranslationally during one-pot reaction and, more importantly, the integrated Cx43 was functionally active in transport. In addition to considerably lower cytotoxicity (

Land use changes have significant modifications on soil conditions, which is likely to induce alterations in the soil bacterial communities. Little is known about the respective contributions of soil variables to these changes in bacterial communities. For this study, high-throughput sequencing technology was applied to measure the change in bacterial community compositions under the effects of soil variables across three land-use types (i.e., reference, degraded, and agricultural wetlands) in the Napahai plateau. Compared with the reference wetland, a pronounced decrease (1.5-5.3 times) in soil water content, soil organic matter, and total and available nitrogen was observed in degraded and agricultural wetlands. However, a conspicuous increase (1.3-5.7 times) was found for the total and available phosphorus, and potassium. Land use also strongly affected the taxonomic composition of soil bacterial assemblages, changing the normalized ratio of Acidobacteria to Proteobacteia, or to δ-proteobacteia. Soil properties had different contributions to the variations in abundance composition of bacterial community. Soil available phosphorus and potassium were the best predictors for changes in bacterial community composition, explaining 80.9% and 82% of the variations, respectively. In contrast, soil organic matter, carbon/nitrogen, total phosphorus, and total and available nitrogen accounted for 58.7-72.7% of the variations in bacterial community composition. Soil pH (24.6%) and soil water content (40.4%) had a minor contribution. Our data suggested that the compositional alterations of microbial communities following land-use change were likely realized through modifications in the availability of primary soil nutrients in the Napahai plateau wetlands.

A porphyrin dimer (BPD) was introduced for selective detection of aluminum ions (Al3+) in a fully aqueous system. BPD produces a ratiometric emission profile in the near-infrared (NIR) region upon Al3+ complexation. The sensing mechanism involves photoinduced electron transfer (PET) and intramolecular charge transfer (ICT). The binding mode of BPD/Al3+ was fully examined based on spectroscopic titration, time-resolved fluorescence decay, NMR spectroscopy, mass spectrometry, and IR analyses. In aqueous media, the strong interaction between Lewis acid Al3+ and nitrogen atoms allows BPD to conquer the high energy barriers of the solvent for only Al3+, resulting in the high selectivity and low detection limit (2.65 × 10–8 M) of the probe. Practical applications of BPD for Al3+ monitoring in real samples and fluorescent prints were further investigated with satisfactory results. With its organic solvent-free sensing medium, good biocompatibility, and NIR ratiometric emission properties, the designed BPD probe is appropriate for ratiometric visualization of Al3+ in living cells.

The role of ribavirin (RBV) in the era of direct-acting antivirals (DAA) is not clear, and DAA studies have been largely genotype- and regimen-specific. Using data from the Chronic Hepatitis Cohort Study, we evaluated the role of RBV and increased DAA treatment duration among patients with chronic hepatitis C (HCV) in routine clinical care. We performed multivariable analysis of data from 4133 patients receiving any of the following: sofosbuvir (SOF); daclatasvir + SOF; grazoprevir + elbasvir; paritaprevir/ritonavir + ombitasvir; simeprevir + SOF; and SOF + ledipasvir; SOF + velpatasvir ± voxilaprevir; and glecaprevir + pibrentasvir—all with/ without RBV. Inverse probability treatment weighting was used to adjust for treatment selection bias. Sustained virological response (SVR) was defined by undetectable HCV RNA 12 weeks after end of therapy. The overall SVR rate was 95%. Mean treatment duration was 12 ± 4.5 weeks. The final model included treatment duration and diabetes, as well as the interaction of RBV with previous treatment status (treatment naïve, interferon treatment failure [TF] or previous DAA TF), cirrhosis status, and HCV genotype (GT). Each one-month increment of treatment duration increased odds of SVR by 99% (aOR = 1.99). Diabetes, previous DAA TF, and decompensated cirrhosis significantly reduced odds of SVR. RBV significantly increased the likelihood of SVR among patients with decompensated cirrhosis (aOR = 5.05), previous DAA treatment failure (aOR = 5.43), and GT3 (aOR = 13.28). Among RBV-free regimens, patients with GT3 were less likely to achieve SVR than those with GT1 or 2 (aOR 0.07). Diabetes, decompensated cirrhosis, and prior DAA TF independently reduced the likelihood of SVR. Longer treatment duration increased likelihood of SVR. Conclusion: RBV increased likelihood of SVR among patients with GT3, previous DAA TF, or decompensated cirrhosis.

Although active constituents extracted from plants show robust in vitro pharmacological effects, low in vivo absorption greatly limits the widespread application of these compounds. A strategy of using phyto-phospholipid complexes represents a promising approach to increase the oral bioavailability of active constituents, which is consist of ‘‘label-friendly” phospholipids and active constituents. Hydrogen bond interactions between active constituents and phospholipids enable phospholipid complexes as an integral part. This review provides an update on four important issues related to phyto-phospholipid complexes: active constituents, phospholipids, solvents, and stoichiometric ratios. We also discuss recent progress in research on the preparation, characterization, structural verification, and increased bioavailability of phyto-phospholipid complexes., Graphical abstract Image, graphical abstract

The expression of drought responsive genes were enhanced by N application and they contributed to drought acclimation. The interactive effects of water and nutrient are crucial for plants. The aim of this study was to elucidate how nitrogen (N) status influence drought acclimation of Populus. A two-factorial design consisting of two N levels (adequate-N and low-N) and two watering treatments (drought stress and well-watered) was used, and an integrative investigation was conducted at the anatomical, physiological and molecular levels. Adequate N supply alleviated the adverse effects of drought stress on root growth in poplars, which may increase water uptake under drought. Nitrogen enhanced leaf anatomical changes and stomata adjustment upon drought stress, lead to less water losses and better growth performance under drought stress. The expression levels of phytohormone signaling components and genes responsible for antioxidative systems and secondary metabolites such as phenylpropanoids were promoted by N application. The expression of abscisic acid (ABA) signaling components was induced by drought when soil N was adequate, which participated in stomata regulation and drought acclimation. The expression of indole acetic acid (IAA) signaling components also was enhanced by N application, which participated in anatomical changes of leaves under drought. These adaptive changes at molecular and anatomical levels contributed to drought acclimation in a synergistic way. Under adequate-N condition, nitrogen and carbon metabolism pathways are being recruited to combat drought, and the C-N interaction play a pivotal role in drought acclimation.

Anxiety disorders are the most prevalent psychiatric disorders, but their pathogenic mechanism remains poorly understood. Here, we report that transmembrane protein 74 (TMEM74), which contains two putative transmembrane domains and exhibits high levels of mRNA in the brain, is closely associated with the pathogenesis of anxiety disorders. TMEM74 was decreased in the serum of patients with anxiety and the basolateral amygdaloid nucleus (BLA) in chronic stress mice. Furthermore, genetic deletion of Tmem74 or selective knockdown of Tmem74 in BLA pyramidal neurons resulted in anxiety-like behaviors in mice. Whole-cell recordings in BLA pyramidal neurons revealed lower hyperpolarization-activated cation current (Ih) and greater input resistance and excitability in Tmem74-/- neurons than in wild-type neurons. Accordingly, surface expression of hyperpolarization-activated cyclic nucleotide-gated 1 (HCN1) channels was also lower in the BLA of Tmem74-/- mice. The Ih current blocker ZD7288 mimicked these effects in BLA pyramidal neurons in wild-type mice but not in Tmem74-/- mice. Consistent with the improvement in anxiety-like behaviors, Tmem74 overexpression restored HCN1 channel trafficking and pyramidal neuron excitability in the BLA of Tmem74-/- and chronic stress mice. Mechanistically, we demonstrate that interactions between Tmem74 and HCN1 are physiologically relevant and that transmembrane domain 1 (TM1) is essential for the cellular membrane localization of Tmem74 to enhance Ih. Together, our findings suggest that Tmem74 coupling with HCN1 acts as a critical component in the pathophysiology of anxiety and is a potential target for new treatments of anxiety disorders.

A natural colourant derived from purple‐fleshed sweet potato (PFSP) by ultrasound‐assisted solvent extraction was examined in terms of its anthocyanins and 6‐month storage stability under different conditions: avoid from light at 25 (AL25) and 37 °C (AL37) and under fluorescent light at 25 °C (FL25). This PFSP colourant mainly contained the following peonidin anthocyanins: peonidin 3‐p‐hydroxybenzoyl sophoroside‐5‐glucoside, peonidin 3‐feruloyl sophoroside‐5‐glucoside, peonidin 3‐caffeoyl sophoroside‐5‐glucoside, peonidin dicaffeoyl sophoroside‐5‐glucoside, peonidin 3‐caffeoyl‐p‐hydroxybenzoyl sophoroside‐5‐glucoside and peonidin caffeoyl‐feruloyl sophoroside‐5‐glucoside. The degradation ratios of PFSP anthocyanins (PFSPAs) under AL25, AL37 and FL25 were around 50.7%, 89.0% and 87.0% respectively. The FL25 and AL37 conditions had similar effects on the colourant's total colour change (ΔEab). The degradation kinetics followed the zero‐order reaction models. The PFSPAs were more stable during storage than other anthocyanins because they mainly consist of di‐acylated and double glycosylated forms of peonidin derivatives. These results may promote the application of PFSPAs as a novel natural colourant in food industry.

Our previous work in acute ischemic stroke and TBI models focused on efficacy and pharmacological parameters of Cerebrolysin®. In this prospective, randomized, blinded, placebo-controlled study we compared efficacy of neuropeptide preparations with putative neurotrophic potential to the reference product Cerebrolysin® by assessing functional outcome and lesion volumes after embolic stroke in a rodent model.Male Wistar rats were subjected to embolic right middle cerebral artery occlusion and were treated with: 1) Cognistar® (Cerebroprotein Hydrolysate) 2.5 ml/kg, 2) Cerebrolysat® 2.5 ml/kg, 3) Cortexin® 1.7 mg/kg, 4) Cerebrolysin® 2.5 ml/kg, or 5) 1 ml of saline according to a pre-generated randomization plan. Dosages were defined according to the packet leaflet of the corresponding preparation and were adapted to the animal model as previously described. All enrolled rats received intraperitoneal injections once daily for 10 consecutive days, starting 4 h after occlusion. Functional outcome was assessed once weekly over four weeks by using a battery of behavioral tests. Infarct volume was measured four weeks after occlusion. Generalized Estimation Equations (GEE) was performed to study the treatment effect on overall functional recovery at day 28 (primary outcome), compared to saline controls.Similar functional outcome was observed for saline control, Cognistar®, Cerebrolysat® and Cortexin®; in contrast, a significantly improved neurological outcome was observed with Cerebrolysin® treatment in comparison to saline as well as to the comparator drug treatment (p .002). However, there was no significant difference in lesion volumes between rats treated with either Cortexin® (33.5 ± 1.9%), Cerebrolysat® (28.5 ± 2.4%), Cognistar® (34.7 ± 2.0%), or Cerebrolysin® (26.5 ± 2.3%) compared to saline-treated rats (30.8 ± 2.1%).Among all tested neuropeptide preparations, Cerebrolysin® was the only agent that was associated with a significant improvement of neurological outcome after stroke.

Earthworms (E) and arbuscular mycorrhizal fungi (AM) as the key components of belowground organisms can form highly complex interactions in soil food web. However, little is known about the direction and power of interactive effects on plant growth and soil fertility in nutrient-limited soils. The objectives aimed to explore the interaction of AM (Claroideoglomus etunicatum) and E (Amynthas divergens yunnanensis) and its effecting mechanism on the plant (Photinia fraseri Dress) growth and nutrient availability in the desertification soils in Karst area. A 2 × 2 factor block design was completely randomly arranged for the treatments in a pot experiment. The four treatments were no E and AM, E inoculation, AM inoculation and inoculations with both E and AM. We confirmed the hypotheses tested about a positive effect of combined inoculations and a more pronounced increase in the plant height (12.5–13.4%), base diameter (24–27%), root colonization (50%) and hyphal length density (312%) compared with separate inoculation. Whereas the inoculation with E or AM had an effect only on the plant height, and AM inoculation only on the hyphal length density. The combined inoculations resulted in a greater reduce in soil pH and a higher increase in soil organic matter and readily oxidizable organic carbon. The effect of combined inoculations on the growth of base diameter, root mycorrhizal colonization and hyphal length density was closely associated with the increase in soil organic matter and soil readily oxidizable organic carbon. In contrast, the effect of separate inoculation on plant height was related with the increase in pH, total nitrogen and phosphorus, plant available nitrogen and phosphorus in soils. Earthworms and AM fungi can create different nutritional niches for plants: E had greater increase in N availability, while AM fungi in P availability. Therefore, we conclude that the combined effects have a greater potential in increasing plant growth, through the complementary and interactive mechanisms of E and AM on modifying nutrient availability in the nutrient-poor soils in Karst stony desertification land.

Carboxylesterases (CarEs) usually play critical roles in the detoxification of toxic chemicals and therefore may be involved in insecticide resistance in agricultural pests. Previous work has shown that CarE 001C from Helicoverpa armigera was able to metabolize the isomers of cypermethrin and fenvalerate. In this study, seven mutants of CarE 001C with single amino acid substitution were produced and expressed in the Escherichia coli. Enzyme kinetic analysis indicated that all seven mutations dramatically reduced enzymatic activities toward the generic substrate α-naphthyl acetate, but in vitro metabolism assay showed that two of the mutations, H423I and R322L, significantly improved hydrolase activities toward fenvalerate, with their recorded specific activities being 3.5 and 5.1 nM·s−1·mg −1 proteins, respectively. Further, thermostability assay showed that the stability of one mutant enzyme was enhanced. This study will help us better understand the potential of CarEs in insecticide detoxification and resistance in H. armigera.

Industry is an important pillar of the national economy. Industrial projects are the most complex and difficult projects to control in the construction industry, and major industrial projects are even more complex and difficult to control. Multi-agent coordination is one of the core issues of industrial projects. Based on an analysis of the engineering and construction chains and agent relationships and agent networks of industrial projects, a complex network of the engineering and construction agents of industrial projects is established, and the complex network structural holes theory is applied to study the nonrepeated relationships among agents in industrial projects. Assuming agents are linked through contract relations and the most critical contract index between the agents in the contract amount, through structural hole analysis considering the EPC and PMC model, the aggregate constraint list is obtained, 2D network diagram and 3D network diagram are shown. According to the aggregate constraint value, the EPC contractor with the minimum aggregate constraint value and the project management company with the minimum aggregate constraint value are the critical agent in EPC and PMC model. By analyzing the complex network comprising different models of industrial projects, it is concluded that the characteristics of the agent maintain an advantage in competition, the coordination mechanism of the integration of agent interests, and multi-agent relations are considered to solve the multi-agent coordination problem in major industrial projects.

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that can cause liver diseases in the host, including hepatitis and hepatomegaly. High mobility group box 1 (HMGB1) is the main inflammatory mediator causing cell injury or necrosis. HMGB1 binds to toll like receptor 4 (TLR4), then activates the nuclear factor-κB (NF-κB) signaling pathway, which promotes the release of inflammatory factors. Our previous studies showed that HMGB1 mediated TLR4/NF-κB signaling pathway plays an important role in liver injury induced by T. gondii infection. Resveratrol (RSV) is a small polyphenol, which has anti-inflammatory, anti-cancer, anti-T. gondii effect. However, the effect of RSV on liver injury caused by T. gondii infection is unclear. This study used the RH strain tachyzoites of T. gondii to infect murine liver line, NCTC-1469 cells to establish an in vitro model and acute infection of mice for the in vivo model to explore the protective effect of RSV on liver injury induced by T. gondii infection. The results showed that RSV inhibited the proliferation of T. gondii in the liver, reduced the alanine aminotransferase/aspartate aminotransferase levels and pathological liver damage. Additionally, RSV inhibited the production of tumor necrosis factor-α, inducible nitric oxide synthase and HMGB1 by interfering with the TLR4/NF-κB signaling pathway. These results indicate that RSV can protect liver injury caused by T. gondii infection by intervening in the HMGB1/TLR4/NF-κB signaling pathway. This study will provide a theoretical basis for RSV treatment of T. gondii infection induced liver injury.

Cerebrolysin has been shown to promote neurovascular protection and repair in preclinical models of stroke and neural injury and is demonstrating promise for stroke and neural injury therapeutic application in the clinic. The effect of Cerebrolysin on the human cerebral endothelial cell function has not been investigated. Using an in-vitro cerebral endothelial cell permeability assay and western blot analyses of tight junction and proinflammatory and procoagulant proteins, the present study showed that tissue plasminogen activator (tPA) and fibrin substantially impaired human cerebral endothelial cell barrier function and increased permeability, which persisted for at least 24 h. western blot analysis revealed that tPA and fibrin significantly increased proinflammatory and procoagulation proteins of intercellular adhesion molecule 1, high mobility group box 1, tumor necrosis factor α and phosphorylated nuclear factor kappa B-p65, and significantly reduced tight junction proteins zonular 1, occludin and claudin. However, Cerebrolysin significantly diminished and reversed tPA- and fibrin-impaired endothelial cell permeability, which was associated with significant reductions of tPA- and fibrin-augmented proinflammatory and procoagulation proteins and significant elevations of tPA- and fibrin-decreased tight junction proteins. The beneficial effect of Cerebrolysin appears specific because cerebroprotein hydrolysate, with a distinct peptide composition, failed to show the reduction of tPA- and fibrin-impaired permeability. These data indicate that cererbrolysin has a therapeutic effect on tPA- and fibrin-impaired cerebral endothelial cell permeability by reducing proinflammatory and procoagulation proteins and by elevating tight junction proteins.

The family of voltage-gated potassium Kv2 channels consists of the Kv2.1 and Kv2.2 subtypes. Kv2.1 is constitutively highly phosphorylated in neurons and its function relies on its phosphorylation state. Whether the function of Kv2.2 is also dependent on its phosphorylation state remains unknown. Here, we investigated whether Kv2.2 channels can be phosphorylated by protein kinase C (PKC) and examined the effects of PKC-induced phosphorylation on their activity and function. Activation of PKC inhibited Kv2.2 currents and altered their steady-state activation in HEK293 cells. Point mutations and specific antibodies against phosphorylated S481 or S488 demonstrated the importance of these residues for the PKC-dependent modulation of Kv2.2. In layer II pyramidal neurons in cortical slices, activation of PKC similarly regulated native Kv2.2 channels and simultaneously reduced the frequency of action potentials. In conclusion, this study provides the first evidence to our knowledge that PKC-induced phosphorylation of the Kv2.2 channel controls the excitability of cortical pyramidal neurons.

Sox9 is important for multiple aspects of development, such as testis, pancreas and heart development. Previous studies have reported that Sox9 induced epithelial‑mesenchymal transition (EMT) and extracellular matrix (ECM) production in organ fibrosis and associated diseases, such as vascular calcification. However, to the best of our knowledge, the role and underlying mechanism of action of Sox9 in renal fibrogenesis remains unknown. The results of the present study revealed that Sox9 expression levels were upregulated in the tubular epithelial cells of a rat model of obstructive nephropathy. Furthermore, the overexpression of Sox9 in NRK‑52E cells was discovered to promote renal tubular EMT and ECM aggregation, and these fibrogenic actions were potentiated by TGF‑β1. Notably, RNA‑sequencing analysis indicated the possible regulatory role of the PI3K/AKT signaling pathway in Sox9‑mediated renal tubular EMT and ECM aggregation. It was further demonstrated that the expression levels of phosphorylated AKT were upregulated in NRK‑52E cells overexpressing Sox9, while the PI3K inhibitors, LY29002 and wortmannin, inhibited the renal tubular EMT and ECM aggregation induced by the overexpression of Sox9 in NEK‑52E cells. In conclusion, the findings of the present study suggested that Sox9 may serve a profibrotic role in the development of renal tubular EMT and ECM aggregation via the PI3K/AKT signaling pathway. Therefore, Sox9 may be considered as a promising target for treating renal fibrosis.

Prolonged occlusion of multiple microvessels causes microvascular injury. G protein-coupled receptor 124 (GPR124) has been reported to be required for maintaining central nervous system (CNS) angiogenesis and blood-brain barrier integrity. However, the molecular mechanisms by which GPR124 regulates pericytes during ischemia have remained elusive. Methods: A microsphere embolism-induced ischemia model was used to evaluate the expression of GPR124 following microsphere embolism. Immunocytochemistry and stochastic optical reconstruction microscopy imaging were used to assess the expression and distribution of GPR124 in human brain vascular pericytes (HBVPs) and after the treatment with 3-morpholino-sydnonimine (SIN-1) or oxygen-glucose deprivation (OGD). The effect of GPR124 knockdown or overexpression on HBVP migration was analyzed in vitro using wound healing assays and a microfluidic device. GPR124 loss-of-function studies were performed in HBVPs and HEK293 cells using CRISPR-Cas9-mediated gene deletion. Time-lapse imaging was used to assess dynamic changes in the formation of filopodia in an individual cell. Finally, to explore the functional domains required for GPR124 activity, deletion mutants were constructed for each of the N-terminal domains. Results: GPR124 expression was increased in pericytes following microsphere embolism. Morphological analysis showed localization of GPR124 to focal adhesions where GPR124 bound directly to the actin binding protein vinculin and upregulated Cdc42. SIN-1 or OGD treatment redistributed GPR124 to the leading edges of HBVPs where GPR124 signaling was required for pericyte filopodia formation and directional migration. Partial deletion of GPR124 domains decreased SIN-1-induced filopodia formation and cell migration. Conclusion: Taken together, our results provide the first evidence for a role of GPR124 in pericyte migration under ischemic conditions and suggest that GPR124 was essential for Cdc42 activation and filopodia formation.

Curcumin, as a high effect and low toxicity anti-cancer drug and photosensitiser, has synergistic and complementary effects with photodynamic therapy (PDT). However, due to its unclear mechanism, PDT's application and efficacy were limited. Notch signaling pathway, which is highly correlates with carcinogenesis and development of cervical cancer, could be a potential therapeutic targets to improve the effectiveness of PDT. Therefore, in this study, we explored the effects of Notch signaling pathway in cervical cancer by curcumin mediated PDT with/without Notch receptor blocker (DAPT), and hope to elucidate its mechanism. Firstly, the effect on the proliferation of cervical cancer Me180 cells were detected with MTT assay, and apoptosis were detected with Annexin V-FITC/PI combined with flow cytometry. Secondly, after establishment of nude mice model, dividing the experimental animals into model group, curcumin PDT group, simple DAPT group, and curcumin-PDT+DAPT group, and analyzing tumor volume changes as well as HE staining in each group. mRNA and protein expression of gene Notch-1 and its downstream NF-κB and VEGF were observed with RT-PCR, immunohistochemical staining and Western-blot with/without inhibition of Notch signaling pathway by DAPT, both in vivo and in vitro experiments. We found both DAPT and curcumin-PDT can inhibit the proliferation and induce apoptosis of cervical cancer cell. The two have synergistic effect in vitro and in vivo. This effect can effectively block the conduction of Notch signaling pathway, which is associated with down-regulation of the expression of Notch1 and NF-κB. Notch signaling pathway could be one of the targets of curcumin-PDT photodynamic therapy.

Engineered nanoparticles have provided a basis for innovative agricultural applications, specifically in plant disease management. In this interdisciplinary study, by conducting comparison studies using macroscale magnesium oxide (mMgO), we evaluated the fungicidal activity of MgO nanoparticles (nMgO) against soilborne Phytophthora nicotianae and Thielaviopsis basicola for the first time under laboratory and greenhouse conditions. In vitro studies revealed that nMgO could inhibit fungal growth and spore germination and impede sporangium development more efficiently than could macroscale equivalents. Indispensably, direct contact interactions between nanoparticles and fungal cells or nanoparticle adsorption thereof were found, subsequently provoking cell morphological changes by scanning electron microscopy/energy-dispersive spectrometry (SEM/EDS) and transmission electron microscopy (TEM). In addition, the disturbance of the zeta potential and accumulation of various modes of oxidative stress in nMgO-exposed fungal cells accounted for the underlying antifungal mechanism. In the greenhouse, approximately 36.58 and 42.35% decreases in tobacco black shank and black root rot disease, respectively, could testify to the efficiency by which 500 μg/ml of nMgO suppressed fungal invasion through root irrigation (the final control efficiency reached 50.20 and 62.10%, respectively) when compared with that of untreated controls or mMgO. This study will extend our understanding of nanoparticles potentially being adopted as an effective strategy for preventing diversified fungal infections in agricultural fields.

Abnormal O-linked-N-acetylglucosamine (O-GlcNAc) concentrations have been associated with a variety of diseases (e.g., cancer, Alzheimer's disease, cardiovascular disease, etc.). However, O-GlcNAc detection is complicated, time-consuming and has poor specificity, therefore, the accurate detection of O-GlcNAc is difficult. In this study, an accurate and sensitive surface plasmon resonance (SPR) biosensor for O-GlcNAc detection that is based on β-D-N-acetylglucosaminidase (OGA) and Au nanoparticles (AuNPs) was developed. In this strategy, AuNPs were used to amplify the SPR signal and improve the biosensor's sensitivity; OGA was used to cleave O-GlcNAc from O-GlcNAcylated biomolecules. The interaction between AuNPs labeled wheat germ agglutinin (AuNPs/WGA) and O-GlcNAcylated biomolecules on a modified Au film treated with and without OGA was recorded by SPR. The change of the SPR signal moves linearly with the amount of O-GlcNAc on the Au film and thus could be used for the detection of O-GlcNAc. By recording the difference of the SPR signals, this method can avoid disturbances from other sugars and nonspecific adsorption of AuNPs and thus enable the accurate detection of O-GlcNAc. The accurate detection range of O-GlcNAc was 4.65 × 10−12 to 4.65 × 10−7 M which was obtained by quantifying the amount of a standard O-GlcNAcylated peptide (O-GlcNAc-CREB), and the detection limit is 4.65 × 10−13 M. More importantly, the strategy was successfully used to detect O-GlcNAc in a real α-crystallin protein, cancer cell lysates and blood samples with satisfactory results. The study's results imply that this accurate and sensitive method has the potential to be applied in the early clinical diagnosis of O-GlcNAc-related diseases.

OBJECTIVECerebrolysin is a neuropeptide preparation that mimics the properties of neurotrophic factors and has had beneficial effects in the treatment of neurodegenerative diseases, stroke, and traumatic brain injury (TBI). To further evaluate treatment schemes, the authors assessed the dose-response of Cerebrolysin on functional improvement in a rat model of mild TBI (mTBI).METHODSThis dose-response study was a prospective, randomized, blinded, and placebo-controlled preclinical experiment. Male Wistar adult rats, subjected to mTBI induced by a closed head impact, were treated randomly with 0 (saline as placebo), 0.8, 2.5, or 7.5 ml/kg of Cerebrolysin 4 hours after mTBI and daily for a total of 10 consecutive days. A battery of cognitive and sensorimotor functional tests was performed over 90 days.RESULTSThe primary outcome was functional improvement over the 90 days; animal weight and death were the secondary and safety outcomes, respectively. A significant (p < 0.001) dose effect of Cerebrolysin on cognitive recovery 3 months after injury was found. Cerebrolysin at a dose of ≥ 0.8 ml/kg significantly (p < 0.001) improved cognitive outcome. The higher dose (7.5 ml/kg) resulted in significantly better cognitive recovery than the lowest doses (0.8 ml/kg) but not relative to the 2.5-ml/kg dose. Cerebrolysin at a dose of 2.5 or 7.5 ml/kg also caused different onset times of significant improvement in sensorimotor function. No differences in body weight or mortality rate among the groups were found.CONCLUSIONSThis preclinical randomized, placebo-controlled, and blinded study with a clinically relevant treatment scheme revealed that Cerebrolysin at doses of 0.8–7.5 ml/kg, administered 4 hours after mTBI and then once daily for a total of 10 consecutive days, improved functional outcomes 3 months after injury. A dose of 2.5 ml/kg is likely an optimal dose for the treatment of experimental mTBI.

Aims Autism spectrum disorder (ASD) is a wide range of neurodevelopmental disorders involving deficits in social interaction and communication. Unfortunately, autism remains a scientific and clinical challenge owing to the lack of understanding the cellular and molecular mechanisms underlying it. This study aimed to investigate the pathophysiological mechanism underlying leukocyte-endothelial adhesion in autism-related neurovascular inflammation. Methods Male BTBR T+tf/J mice were used as an autism model. The dynamic pattern of leukocyte-endothelial adhesion in mouse cerebral vessels was detected by two-photon laser scanning microscopy (TPLSM). Using FACS, RT-PCR, and Western blotting, we explored the expression of cell adhesion molecules, the mRNA expression of endothelial chemokine, the protein levels of cathepsin B, and inflammatory mediators. Results We found a significant increase in leukocyte-endothelial adhesion in BTBR mice, accompanied by elevated expression of the adhesion molecule neutrophils CD11b and endothelial ICAM-1. Our data further indicate that elevated neutrophil cathepsin B levels contribute to elevated endothelial chemokine CXCL7 levels in BTBR mice. The pharmacological inhibition of cathepsin B reverses the enhanced leukocyte-endothelial adhesion in the cerebral vessels of autistic mice. Conclusion Our results revealed the prominent role of cathepsin B in modulating leukocyte-endothelial adhesion during autism-related neurovascular inflammation and identified a promising novel approach for autism treatment.