Activation of Wnt signaling reduces high-glucose mediated damages on skin fibroblast cells

Objective(s): High-glucose (HG) stress, a mimic of diabetes mellitus (DM) in culture cells, alters expression of a large number of genes including Wnt and NF-κB signaling-related genes; however, the role of Wnt signaling during HG-mediated fibroblast damage and the relationship between Wnt and NF-κB signaling have not been understood. In this study, we aimed to investigate the ffects of Wnt signaling on HG-mediated damages. Materials and Methods: Wnt3a was treated to HG-stressed human primary foreskin fibroblasts and the levels of Wnt signaling markers and cell proliferation were monitored. In addition, Wnt3a and NF-κB signaling inhibitor were assisted to analyze the relationship between two pathways. Results: The results indicated that HG treatment repressed β-catenin level, and Wnt3a treatment increased the levels of β-catenin and FZD8 as well as cell proliferation. RNA-seq based transcriptome analysis identified 207 up-regulated and 200 down-regulated genes upon Wnt3a supply. These altered genes are distributed into 20 different pathways. In addition, gene ontology (GO) analysis indicates that 20 GO terms are enriched. Wnt signaling genes were further verified by qRT-PCR and the results were similar with RNA-seq assay. Since NF-κB signaling negatively regulates Wnt marker gene expression, Bay117082, a typical NF-κB signaling inhibitor and Wnt3a were supplemented for testing β-catenin and phosphorylated IκBα (p-IκBα), respectively. Conclusion: HG positively inhibits Wnt signaling, and signaling activation via supplementation of Wnt3a rescued the defect caused by HG. NF-κB signaling negatively regulates accumulation of β-catenin, but Wnt signaling has no effects on IκBα activation

TiAVox: Time-aware Attenuation Voxels for Sparse-view 4D DSA Reconstruction

Four-dimensional Digital Subtraction Angiography (4D DSA) plays a critical role in the diagnosis of many medical diseases, such as Arteriovenous Malformations (AVM) and Arteriovenous Fistulas (AVF). Despite its significant application value, the reconstruction of 4D DSA demands numerous views to effectively model the intricate vessels and radiocontrast flow, thereby implying a significant radiation dose. To address this high radiation issue, we propose a Time-aware Attenuation Voxel (TiAVox) approach for sparse-view 4D DSA reconstruction, which paves the way for high-quality 4D imaging. Additionally, 2D and 3D DSA imaging results can be generated from the reconstructed 4D DSA images. TiAVox introduces 4D attenuation voxel grids, which reflect attenuation properties from both spatial and temporal dimensions. It is optimized by minimizing discrepancies between the rendered images and sparse 2D DSA images. Without any neural network involved, TiAVox enjoys specific physical interpretability. The parameters of each learnable voxel represent the attenuation coefficients. We validated the TiAVox approach on both clinical and simulated datasets, achieving a 31.23 Peak Signal-to-Noise Ratio (PSNR) for novel view synthesis using only 30 views on the clinically sourced dataset, whereas traditional Feldkamp-Davis-Kress methods required 133 views. Similarly, with merely 10 views from the synthetic dataset, TiAVox yielded a PSNR of 34.32 for novel view synthesis and 41.40 for 3D reconstruction. We also executed ablation studies to corroborate the essential components of TiAVox. The code will be publically available.Comment: 10 pages, 8 figure

Development and Validation of LC-MS/MS Method for Determination of Ondansetron in rat Plasma and its Application

A selective and sensitive liquid chromatography-mass spectrometry (LC–MS) method for determination of ondansetron in rat plasma was developed and validated. After addition of midazolam as internal standard (IS), protein precipitation by acetonitrile was used as sample preparation, and chromatography involved Agilent SB-C18 column (2.1 x 150 mm, 5 μm) using 0.1 % formic acid in water and acetonitrile as a mobile phase with gradient elution. Detection involved positive ion mode electrospray ionization (ESI), and multiple reaction monitoring (MRM) mode was used for quantification of target fragment ions m/z 294.0→169.7 for ondansetron and m/z 326.0→291.0 for midazolam (internal standard, IS). The assay was linear over the range of 5–1000 ng/mL for ondansetron, with a lower limit of quantitation (LLOQ) of 5 ng/mL for ondansetron. Intra- and inter-day precisions were less than 14 % and the accuracies were in the range of 94.7-113.5 % for ondansetron. This developed method was successfully applied for the determination of ondansetron in rat plasma for pharmacokinetic study.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Determination of ramosetron in rat plasma by LC-ESI-MS and its application

A sensitive and simple liquid chromatography/electrospray mass spectrometry (LC-ESI-MS) method for determination of ramosetron in rat plasma using one-step protein precipitation was developed and validated. After addition of midazolam as internal standard (IS), protein precipitation by acetonitrile was used as sample preparation. Chromatographically separation was achieved on an SB-C18 (2.1 mm × 150 mm, 5 μm) column with acetonitrile-0.1 % formic acid as the mobile phase with gradient elution. Electrospray ionization (ESI) source was applied and operated in positive ion mode; selected ion monitoring (SIM) mode was used to quantification using target fragment ions m/z 280 for ramosetron and m/z 326 for the IS. Calibration plots were linear over the range of 10-1000 ng/mL for ramosetron in rat plasma. Lower limit of quantification (LLOQ) for Ramosetron was 10 ng/mL. Mean recovery of ramosetron from plasma was in the range of 88.5-92.8 %. CV of intra-day and inter-day precision were both less than 15 %. This method is simple and sensitive enough to be used in pharmacokinetic study for determination of ramosetron in rat plasma.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Improved photocatalytic activity of g-C3N4 derived from cyanamide-urea solution

This paper describes the fabrication of g-C3N4 by the polymerization of cyanamide-urea solution at elevated temperatures. The textural properties and electronic band structure of the obtained g-C3N4 were investigated in detail. The photocatalytic activity for both oxidative and reductive reactions of the as-synthesized g-C3N4 was found to be enhanced as the polymerization temperature increase and the g-C3N4 obtained at 700 degrees C (CN-700) showed the best photocatalytic activity under visible-light (lambda > 420 nm). Considering that the rather wide band gap (3.01 eV) of CN-700 disables the electron transition from the valence band to the conduction band by visible light (l > 420 nm), it is believed the n-pi* transition, which is alternatively proposed in this study, plays a key role in its photocatalytic activity. In light of this discovery, the variation of the electron-transition mechanism for g-C3N4 fabricated at different polymerization temperatures has been firstly investigated

Application of citrate as a tricarboxylic acid (TCA) cycle intermediate, prevents diabetic-induced heart damages in mice

Objective(s):Higher cellular reactive oxygen species (ROS) levels is important in reducing cellular energy charge (EC) by increasing the levels of key metabolic protein, and nitrosative modifications, and have been shown to damage the cardiac tissue of diabetic mice. However, the relation between energy production and heart function is unclear. Materials and Methods:Streptozotocin (STZ, 150 mg/kg body weight) was injected intraperitoneally once to mice that had been fasted overnight for induction of diabetes. After diabetic induction, mice received citrate (5 µg/kg) through intraperitoneal injection every other day for 5 weeks. The caspase-3, plasminogen activator inhibitor 1 (PAI1), protein kinase B (PKB), commonly known as AKT and phosphorylated-AKT (p-AKT) proteins were examined to elucidate inflammation and apoptosis in the heart. For histological analysis, heart samples were fixed with 10% formalin and stained with hematoxylin-eosin (HE) and Sirius red to assess pathological changes and fibrosis. The expression levels[AGA1]  of marker proteins, tyrosine nitration, activity of ATP synthase and succinyl-CoA:3-ketoacid coenzyme A transferase-1 (SCOT), and EC were measured. Results:Intraperitoneal injection of citrate significantly reduced caspase-3 and PAI-1 protein levels and increased p-AKT level on the 5th week; EC in the heart was found to be increased as well. Further, the expression level, activity, and tyrosine nitration of ATP synthase and SCOT were not affected after induction of diabetes. Conclusion: Results indicate that application of citrate, a tricarboxylic acid (TCA) cycle intermediate, might alleviate cardiac dysfunction by reducing cardiac inflammation, apoptosis, and increasing cardiac EC

Genetic Variation of SARS Coronavirus in Beijing Hospital

To characterize genetic variation of severe acute respiratory syndrome–associated coronavirus (SARS-CoV) transmitted in the Beijing area during the epidemic outbreak of 2003, we sequenced 29 full-length S genes of SARS-CoV from 20 hospitalized SARS patients on our unit, the Beijing 302 Hospital. Viral RNA templates for the S-gene amplification were directly extracted from raw clinical samples, including plasma, throat swab, sputum, and stool, during the course of the epidemic in the Beijing area. We used a TA-cloning assay with direct analysis of nested reverse transcription–polymerase chain reaction products in sequence. One hundred thirteen sequence variations with nine recurrent variant sites were identified in analyzed S-gene sequences compared with the BJ01 strain of SARS-CoV. Among them, eight variant sites were, we think, the first documented. Our findings demonstrate the coexistence of S-gene sequences with and without substitutions (referred to BJ01) in samples analyzed from some patients

Community perspectives of flagship species: can conservation motivators mitigate human-wildlife conflict?

Public perception of endangered species is crucial for successful management of community-based conservation and sustainability of national parks. By the method of choice experiment, our study evaluated conservation preferences and willingness to donate money for flagship and non-flagship species using a choice experiment with 409 residents living near the Lanstang river source of Sanjiangyuan National Park, China. We found that flagship species such as the Snow leopard (Pristine plateau) and White-lipped deer (Przewalskium albirostris) generated more conservation funds than non-flagship species. However, not all flagship species were accepted. Respondents disliked Tibetan brown bears (Ursus arctos pruinosus) due to direct human-wildlife conflicts such as bodily injury and property damage. Heterogeneity of preference was influenced by household income, religious beliefs, ethnicity, culture, and conservation awareness. Results can be used to establish a local community-participative framework by combining conservation motivations that alleviate human-wildlife conflict

Circulating methylation level of HTR2A is associated with inflammation and disease activity in rheumatoid arthritis

ObjectivesHTR2A is previously identified as a susceptibility gene for rheumatoid arthritis (RA). In this study, we performed the association analysis between DNA methylation of HTR2A with RA within peripheral blood samples.MethodsWe enrolled peripheral blood samples from 235 patients with RA, 30 osteoarthritis (OA) patients, and 30 healthy controls. The DNA methylation levels of about 218 bp from chr13: 46898190 to chr13: 46897973 (GRCh38/hg38) around HTR2A cg15692052 from patients were analyzed by targeted methylation sequencing.ResultsWe measured methylation status for 7 CpGs in the promoter region of HTR2A and obseved overall methylation status are signficantly increased in RA compared with normal inviduals (FDR= 9.05 x 10-5). The average cg15692052 methylation levels (methylation score) showed a positive correlation with CRP (r=0.15, P=0.023). Compared with the OA group or HC group, the proportion of haplotypes CCCCCCC (FDR=0.02 and 2.81 x 10-6) is signficantly increased while TTTTTCC (FDR =0.01) and TTTTTTT(FDR =6.92 x 10-3) are significantly decreased in RA. We find methylation haplotypes combining with RF and CCP could signficantly enhance the performance of the diagnosing RA and its comorbidities (hypertension, interstitial lung disease, and osteoporosis), especially in interstitial lung disease.ConclusionsIn our study, we found signficant hypermethylation of promoter region of HTR2A which indicates the potential clinical diagnostic role in rheumatoid arthritis

A comprehensive review of Tripterygium wilfordii hook. f. in the treatment of rheumatic and autoimmune diseases: Bioactive compounds, mechanisms of action, and future directions

Rheumatic and autoimmune diseases are a group of immune system-related disorders wherein the immune system mistakenly attacks and damages the body’s tissues and organs. This excessive immune response leads to inflammation, tissue damage, and functional impairment. Therapeutic approaches typically involve medications that regulate immune responses, reduce inflammation, alleviate symptoms, and target specific damaged organs. Tripterygium wilfordii Hook. f., a traditional Chinese medicinal plant, has been widely studied in recent years for its application in the treatment of autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis. Numerous studies have shown that preparations of Tripterygium wilfordii have anti-inflammatory, immunomodulatory, and immunosuppressive effects, which effectively improve the symptoms and quality of life of patients with autoimmune diseases, whereas the active metabolites of T. wilfordii have been demonstrated to inhibit immune cell activation, regulate the production of inflammatory factors, and modulate the immune system. However, although these effects contribute to reductions in inflammatory responses and the suppression of autoimmune reactions, as well as minimize tissue and organ damage, the underlying mechanisms of action require further investigation. Moreover, despite the efficacy of T. wilfordii in the treatment of autoimmune diseases, its toxicity and side effects, including its potential hepatotoxicity and nephrotoxicity, warrant a thorough assessment. Furthermore, to maximize the therapeutic benefits of this plant in the treatment of autoimmune diseases and enable more patients to utilize these benefits, efforts should be made to strengthen the regulation and standardized use of T. wilfordii