课程文献中心 更多>>

关键词： Autohaemotherapy   Gold therapy   Corneal wound healing   Human corneal epithelial cells   EPIDERMAL-GROWTH-FACTOR   INTERLEUKIN-6   EXPRESSION   MIGRATION   ALPHA   NANOPARTICLES   MMP-9   RNA  

摘要： Background: Wound healing disorders characterised by impaired or delayed reepithelialisation are a serious medical problem. In the present study, we show that gold-based blood serum therapy is a suitable therapeutic approach and shows a supportive effect in wound closure of human corneal epithelial cells (HCE) in primary in vitro experiments. Methods: For this purpose, blood from healthy individuals was incubated without (***) or with gold-microparticles (***) for 24 h. Prior to human epithelial cell stimulation (HCE), the gold particles were removed and the serum was diluted in DMEM (10 % or 30 %). Both groups of serum were compared after injury. HCE were cultured and injured (corneal in vitro wound model) and then stimulated with *** or ***. Results: Treatment with serum from a gold-based serum therapy (***) shows a supportive effect on wound healing in HCE cells in vitro. In addition, gold therapy supports the secretion of important cytokines normally associated with ocular surface wound healing (IL-1 beta, IL-6, TNF-alpha and TGF-beta) in HCE cells. Conclusions: Therapy with gold-based blood serum significantly promotes the secretion and expression of cytokines and growth factors in HCE cells in vitro. Further preclinical experiments are necessary to demonstrate the influence of this therapy on HCE cells for possible clinical application on the human ocular surface and to prove its function also in poorly healing corneal lesions. (C) 2021 Elsevier GmbH. All rights reserved.

关键词： COVID-19   SARS-CoV-2   Spike protein   Binding free energy   Molecular dynamics simulation  

摘要： SARS-CoV-2 is the causative agent of the ongoing viral pandemic of COVID-19. After the emergence of this virus, it became a global public health concern and quickly evolved into a pandemic. Mexico is currently in the third position in the number of deaths due to SARS-CoV-2. To date, there have been several lineages of SARS-CoV-2 worldwide; in the Mexican population, two variants of the spike protein (S-protein) are found, localized at H49Y and D614G, which have been related to increased infectivity with respect to the wild-type S-protein. To understand how these differences impact the structural behavior of the S-protein of SARS-CoV-2, as well as binding with ACE2, we performed MD simulations combined with the molecular mechanics generalized Born/PoissonBoltzmann surface area (MMGB(PB)SA) approach starting from X-ray crystallography data. Energetic and structural analysis showed that the differences in infectivity can be explained by differences in affinity of the protein-protein interface between the wild-type and mutant S-protein with ACE2. Conformational analysis showed that molecular recognition between the S-protein and ACE2 is linked to a decrease in the conformational flexibility of wild-type and mutant S-protein; however, an increase in the conformational mobility of ACE2 could also contribute to the binding affinity observed using the MMGB(PB)SA method.

关键词： Parkinson’s disease   Human amniotic epithelial cells   Neuroprotection   Dopaminergic neurons   Anti-inflammatory factors   Anti-oxidative stress   CILIARY NEUROTROPHIC FACTOR   STEM-CELLS   DOPAMINERGIC-NEURONS   RAT MODEL   GROWTH-FACTOR   OXIDATIVE STRESS   SUBSTANTIA-NIGRA   ANIMAL-MODELS   TRANSPLANTATION   BRAIN  

摘要： Human amniotic epithelial cells (hAECs) have been reported to have neuroprotective roles in Parkinson's disease (PD) animal models. However, the molecular mechanism is not fully understood. The present study was designed to explore the possible mechanism by which hAECs ameliorate PD symptoms and the important paracrine factors produced by hAECs that attribute to the recovery of dopaminergic neurons. Thus, we performed in vivo and in vitro experiments with hAECs in PD models or lesioned dopaminergic neurons, respectively. First, hAECs were transplanted into the striatum of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mice and motor deficits were significantly attenuated. Second, the grafts prevented the loss of nigral dopaminergic neurons and promoted the outgrowth of neurites and striatal axon fibers in PD mice. In addition, decreased microglial activation, inflammatory factor levels and MPTP-induced excessive reactive oxygen species (ROS) levels were also observed in hAEC-treated PD mice. In vitro, we found that the conditioned medium (CM) from hAECs promoted the survival of mesencephalic dopaminergic neurons stimulated with 1-methyl-4-phenylpyridine (MPP+) and induced neurite outgrowth. Next, analysis of hAEC-CM with an antibody array of 507 soluble target proteins revealed that the levels of many neurotrophic factors, growth factors, neuronal cell adhesion molecule (NrCAM) and anti-inflammatory factors were evidently high. In addition, antibody neutralization experiments showed that many of these factors contributed to the survival and growth of dopaminergic neurons and neurite outgrowth. More importantly, we found that the anti-inflammatory factor interleukin-1 receptor antagonist (IL-1ra) also augmented the survival of dopaminergic neurons, demonstrating for the first time an anti-oxidative and anti-inflammatory role of hAECs in PD mice, which represents a novel molecular mechanism of hAECs in the treatment of *** molecular mechanism of hAE

关键词： Arsenic   In vitro   Genomics   Mode of action   Carcinogenicity   NF-KAPPA-B   GENE-EXPRESSION CHANGES   DOSE-RESPONSE   24-HR EXPOSURES   DRINKING-WATER   ACTIVATION   CANCER   MDM2   CARCINOGENICITY   PATHWAYS  

摘要： Evidence from both in vivo and in vitro studies suggests that gene expression changes from long-term exposure to arsenite evolve markedly over time, including reversals in the direction of expression change in key regulatory genes. In this study, human uroepithelial cells from the ureter segments of 4 kidney-donors were continuously treated in culture with arsenite at concentrations of 0.1 or 1 mu M for 60 days. Gene expression at 10, 20, 30, 40, and 60 days was determined using Affymetrix human genome microarrays and signal pathway analysis was performed using GeneGo Metacore. Arsenic treated cells continued to proliferate for the full 60-day period, whereas untreated cells ceased proliferating after approximately 30 days. A peak in the number of gene changes in the treated cells compared to untreated controls was observed between 30 and 40 days of exposure, with substantially fewer changes at 10 and 60 days, suggesting remodeling of the cells over time. Consistent with this possibility, the direction of expression change for a number of key genes was reversed between 20 and 30 days, including CFOS and MDM2. While the progression of gene changes was different for each subject, a common pattern was observed in arsenic treated cells over time, with early upregulation of oxidative stress responses (HMOX1, NQ01, TXN, TXNRD1) and down-regulation of immune/inflammatory responses (IKK alpha). At around 30 days, there was a transition to increased inflammatory and proliferative signaling (AKT, CFOS), evidence of epithelial-to-mesenchymal transition (EMT), and alterations in DNA damage responses (MDM2, ATM). A common element in the changing response of cells to arsenite over time appears to involve up-regulation of MDM2 by inflammatory signaling (through AP-1 and NF-kappa B), leading to inhibition of P53 function.

关键词： Thyroid gland   T lymphocytes   Lymphoproliferative disorders   Hashimoto disease   Tertiary lymphoid structures   PRIMARY LYMPHOMA   GLAND  

摘要： Thyroid lymphoproliferative disorder is known to be associated with Hashimoto thyroiditis. The major histological types of thyroid lymphoproliferative disorders are malignant lymphoma, such as diffuse large B cell lymphoma, extranodal marginal zone B cell lymphoma of mucosa-associated lymphoid tissue (MALT), and follicular lymphoma. All of these are B cell lymphomas; however, T cell lymphoproliferative disorders represent only a minority of cases. Since T cell lymphoproliferative disorder rarely develops and T cell lymphoma is a heterogeneous entity, it is difficult to concisely summarize this complex topic. Here, we present a case of primary thyroid T cell lymphoproliferative disorder reminiscent of indolent T cell lymphoproliferative disorder of the gastrointestinal tract: the pathological findings from this case also were similar to those of MALT lymphoma such as small lymphoid cells, lymphoepithelial lesions, and a low Ki-67 labeling index. T cell lymphoproliferative disorders with these pathological features have been reported in the thyroid gland, lymph nodes, palate, spleen, and gastrointestinal tract. Furthermore, we reviewed and summarized the studies on thyroid T cell lymphoproliferative disorders and compared the cases and their clinicopathological characteristics.

关键词： HEREDITARY HEMORRHAGIC TELANGIECTASIA   FACTOR RECEPTOR-2 EXPRESSION   SP1-DEPENDENT DNA-BINDING   TGF-BETA RECEPTOR   SHEAR-STRESS   GENE   ANGIOGENESIS   MUTATIONS   BEVACIZUMAB   ANGIOPOIETIN-1  

摘要： Hereditary hemorrhagic telangiectasia (HHT) is a genetic disease characterized by vascular dysplasia. Mutations of the endoglin (ENG) gene that encodes a co-receptor of the transforming growth factor beta 1 signaling pathway cause type I HHT. ENG is primarily expressed in endothelial cells (ECs), but its interaction with other key angiogenic pathways to control angiogenesis has not been well addressed. The aim of this study is to investigate ENG interplay with VEGFR2, FGFR1 and TIE2 in primary human ECs. ENG was knocked-down with siRNA in human umbilical vein ECs (HUVECs) and human lung microvascular ECs (HMVEC-L). Gene expression was measured by RT-qPCR and Western blotting. Cell signaling pathway activation was analyzed by detecting phosphor-ERK and phosphor-AKT levels. Cell migration and apoptosis were assessed using the Boyden chamber assay and the CCK-8 Kit, respectively. Loss of ENG in HUVECs led to significantly reduced expression of VEGFR2 but not TIE2 or FGFR1, which was also confirmed in HMVEC-L. HUVECs lacking ENG had significantly lower levels of active Rac1 and a substantial reduction of the transcription factor Sp1, an activator of VEGFR2 transcription, in nuclei. Furthermore, VEGF- but not bFGF- or angiopoietin-1-induced phosphor-ERK and phosphorAKT were suppressed in ENG deficient HUVECs. Functional analysis revealed that ENG knockdown inhibited cell migratory but enhanced anti-apoptotic activity induced by VEGF. In contrast, bFGF, angiopoietin-1 and -2 induced HUVEC migration and anti-apoptotic activities were not affected by ENG knockdown. In conclusion, ENG deficiency alters the VEGF/VEGFR2 pathway, which may play a role in HHT pathogenesis. (Translational Research 2021; 235:129-143)

关键词： CAP2   LEADS  

摘要： Striated muscle is intricately designed to provide efficient and powerful muscle contractions. Recently, a long sought-after missing component of the thin filament pointed- end machinery was discovered: cyclase- associated protein 2 (CAP2). CAP2 was identified as a crucial contributor to actin polymerization, striated muscle development, and severe muscle disease when mutated.

关键词： Antibiotics   Carp   Cell line   Cytotoxicity   Ecotoxicity   Mussel   MUSSEL MYTILUS-EDULIS   WASTE-WATER   DIGESTIVE GLAND   ACUTE TOXICITY   OXOLINIC ACID   ANTIBIOTICS   PHARMACEUTICALS   BIOACCUMULATION   ENVIRONMENT   CULTURE  

摘要： The ubiquitous use of antibiotics leads exposure of these chemicals on non-target aquatic species, while the toxicity assays for these chemicals are time/labor consuming and expensive. Alternative approaches using primary cell cultures which retain the tissue functionality at its highest form have received global attention compared to cell lines. In the current study, the cytotoxic effects of two commonly used antibiotics from amphenicol (florfenicol) and macrolide (erythromycin) groups were evaluated on primary cell cultures of Unio crassus (mantle, digestive gland, gill, and gonad) and Cyprinus carpio (gill and liver) using MTT and Neutral Red assays. The highest cytotoxic effects were found on the mussel digestive gland and carp liver cells for florfenicol and erythromycin, while the lowest cytotoxic effects were found in mussel mantle cells for both drugs in the MTT test. In the NR test, the highest cytotoxic effects of erythromycin and florfenicol were found in the mussel gill, mantle, gonad, and carp gill cells; the lowest cytotoxic effect of erythromycin was found in the mussel digestive gland, while the lowest effect of florfenicol was found in the carp liver cells. The cytotoxicity of florfenicol was quite low for the carp liver, while the cytotoxicity of erythromycin was quite low in the mussel digestive tract. Thus, it was concluded that cells made from mussel tissues could be used in ecotoxicity tests, and sensitivity may vary according to the tissue.

关键词： Bestrophin-1   Best1 channel   DRG neurons   GABA receptors   Primary afferents   Satellite cells   SPINAL-CORD   RELEASE   RECEPTORS   MOTONEURONS  

摘要： Primary afferent fibers express extrasynaptic GABA(A) and GABA(B) receptors in the axons and soma. However, whether these receptors are tonically activated by ambient GABA and the source of the neuro-transmitter is presently unknown. Here, we show that GABA release from dorsal root ganglia (DRG) does not depend on extracellular calcium, but depends upon calcium released from intracellular stores, and is mediated by Best1 channels. Using a preparation consisting of the spinal nerve in continuity with the DRG and the dorsal root, we found that endogenous GABA tonically activates GABA receptors, depressing the excitability of the primary afferents. In addition, using HPLC we found that GABA is released in the DRG, and by immunofluorescence microscopy we show the presence of GABA, the Best1 channel, and some enzymes of the putrescine pathway of GABA biosynthesis, in glutamine synthase- and GFAP-positive satellite glial cells. Last, we found that the blockade of the Best1 channel activity reduced the excitability of primary afferents and prevented the activation of the GABA receptors. These results suggest that satellite glial cells may be the source of endogenous GABA released in the DRG via Best1 channels, which tonically activates extrasynaptic GABA receptors. (C) 2020 Elsevier B.V. and Japan Neuroscience Society. All rights reserved.

关键词： Terrain reconstruction   Remote sensing image   Multispectral image   Convolutional neural network   Material classification   EXTRACTION   AERIAL   RECONSTRUCTION   SEGMENTATION   REGISTRATION   MODELS   ROADS  

摘要： For real-world simulation, terrain models must combine various types of information on material and texture in terrain reconstruction for the three-dimensional numerical simulation of terrain. However, the construction of such models using the conventional method often involves high costs in both manpower and time. Therefore, this study used a convolutional neural network (CNN) architecture to classify material in multispectral remote sensing images to simplify the construction of future models. Visible light (i.e., RGB), near infrared (NIR), normalized difference vegetation index (NDVI), and digital surface model (DSM) images were examined. This paper proposes the use of the robust U-Net (RUNet) model, which integrates multiple CNN architectures, for material classification. This model, which is based on an improved U-Net architecture combined with the shortcut connections in the ResNet model, preserves the features of shallow network extraction. The architecture is divided into an encoding layer and a decoding layer. The encoding layer comprises 10 convolutional layers and 4 pooling layers. The decoding layer contains four upsampling layers, eight convolutional layers, and one classification convolutional layer. The material classification process in this study involved the training and testing of the RUNet model. Because of the large size of remote sensing images, the training process randomly cuts subimages of the same size from the training set and then inputs them into the RUNet model for training. To consider the spatial information of the material, the test process cuts multiple test subimages from the test set through mirror padding and overlapping cropping; RUNet then classifies the subimages. Finally, it merges the subimage classification results back into the original test image. The aerial image labeling dataset of the National Institute for Research in Digital Science and Technology (Inria, abbreviated from the French Institut national de recherche en