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1.
Cell Death Discov ; 10(1): 395, 2024 Sep 05.
Article in English | MEDLINE | ID: mdl-39237520

ABSTRACT

Abdominal aortic aneurysms (AAA) are a significant health concern in developed countries due to their considerable mortality rate. The crucial factor of the progression of AAA is the release of neutrophils and neutrophil extracellular traps (NETs). Magnetic particle imaging (MPI) is a new imaging technique that offers the capability to detect superparamagnetic iron oxide nanoparticles (SPION) with exceptional sensitivity. We aimed to investigate the functional imaging of MPI for the detection and monitoring of neutrophil infiltration within AAA. A novel multimodal imaging agent targeting neutrophils, PEG-Fe3O4-Ly6G-Cy7 nanoparticles (Ly6G NPs), were designed by coupling Fe3O4 nanoparticles with Ly6G antibodies and Cy7. The targeting and sensitivity of Ly6G NPs were assessed using MPI and fluorescence imaging (FLI) in the AAA mouse model. After the inhibition of NETosis, the degree of neutrophil infiltration and AAA severity were assessed using MPI with Ly6G NPs. Ly6G NPs accurately localized and quantitatively analyzed AAA lesion sites in mice using MPI/FLI/CT. Compared to the control group, elevated MPI and FLI signal intensities were detected at the abdominal aortic lesion site, and neutrophil infiltration and NETs accumulation were detected by histological analysis in the AAA models. After the inhibition of NETs accumulation in vivo, pathological damage in the abdominal aorta was significantly reduced, along with a decrease in the accumulation of Ly6G NPs and MPI signals. This multimodal MPI strategy revealed that nanoparticles targeting Ly6G can be used to detect neutrophil infiltration within AAA and monitor AAA severity.

2.
Front Pharmacol ; 15: 1388540, 2024.
Article in English | MEDLINE | ID: mdl-39108744

ABSTRACT

Background: Atherosclerosis (AS) is a chronic arterial pathology and a leading cause of vascular disease-related mortality. Fatty streaks in the arterial wall develop into atherosclerosis and characteristic plaques. Clinical interventions typically involve lipid-lowering medications and drugs for stabilizing vulnerable plaques, but no direct therapeutic agent specifically targets atherosclerosis. Garlic, also locally known as DASUAN, is recognized as a widely sold herbal dietary supplement esteemed for its cardiovascular benefits. However, the specific mechanisms of garlic's anti-atherosclerotic effects remain unclear. Aims: This study aims to elucidate the pharmacological mechanisms through which garlic ameliorates atherosclerosis. Methods: The study identified the major active components and targets of garlic by screening the TCMSP, TCM-ID, and, ETCM databases. Atherosclerosis-associated targets were obtained from the DisGeNET, GeneCards, and DiGSeE databases, and garlic intervention targets were determined through intersection. Utilizing the intersected genes, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using R software. A garlic component-disease target network was constructed using Cytoscape. RNA-seq datasets from the GEO database were utilized to identify differentially expressed genes (DEGs) associated with atherosclerosis. The target genes were intersected with DEGs and the FerrDb (ferroptosis database). Molecular docking predicted the binding interactions between active components and the core targets. In vitro and in vivo experiments validated the identified core targets. Results: The integration of garlic drug targets with atherosclerotic disease targets identified 230 target genes. Intersection with RNA-seq DEGs revealed 15 upregulated genes, including 8 target genes related to ferroptosis. Molecular docking indicated favorable affinities between garlic active components [Sobrol A, (+)-L-Alliin, Benzaldoxime, Allicin] and target genes (DPP4, ALOX5, GPX4). Experimental validation showed that GARLIC reduces the expression of ferroptosis-related genes in AS, suggesting its therapeutic potential through the regulation of ferroptosis. Conclusion: Garlic ameliorates atherosclerosis by targeting intra-plaque ferroptosis and reducing lipid peroxidation. These findings provide novel insights into the pharmacological mechanisms underlying the efficacy of garlic in treating AS.

3.
Waste Manag ; 187: 156-166, 2024 Oct 01.
Article in English | MEDLINE | ID: mdl-39043078

ABSTRACT

Waste electrical and electronic equipment (WEEE) has become a critical environmental problem. Catalytic pyrolysis is an ideal technique to treat and convert the plastic fraction of WEEE into chemicals and fuels. Unfortunately, research using real WEEE remains relatively limited. Furthermore, the complexity of WEEE complicates the analysis of its pyrolytic kinetics. This study applied the Fraser-Suzuki mathematical deconvolution method to obtain the pseudo reactions of the thermal degradation of two types of WEEE, using four different catalysts (Al2O3, HBeta, HZSM-5, and TiO2) or without a catalyst. The main contributor(s) to each pseudo reaction were identified by comparing them with the pyrolysis results of the pure plastics in WEEE. The nth order model was then applied to estimate the kinetic parameters of the obtained pseudo reactions. In the low-grade electronics pyrolysis, the pseudo-1 reaction using TiO2 as a catalyst achieved the lowest activation energy of 92.10 kJ/mol, while the pseudo-2 reaction using HZSM-5 resulted in the lowest activation energy of 101.35 kJ/mol among the four catalytic cases. For medium-grade electronics, pseudo-3 and pseudo-4 were the main reactions for thermal degradation, with HZSM-5 and TiO2 yielding the lowest pyrolytic activation energies of 75.24 and 226.39 kJ/mol, respectively. This effort will play a crucial role in comprehending the pyrolysis kinetic mechanism of WEEE and propelling this technology toward a brighter future.


Subject(s)
Electronic Waste , Plastics , Pyrolysis , Electronic Waste/analysis , Plastics/chemistry , Kinetics , Catalysis , Titanium/chemistry , Models, Theoretical , Refuse Disposal/methods
4.
Genes Nutr ; 19(1): 13, 2024 Jul 23.
Article in English | MEDLINE | ID: mdl-39044161

ABSTRACT

OBJECTIVE: Employing network pharmacology and molecular docking, the study predicts the active compounds in garlic and elucidates their mechanism in inhibiting the development of alcoholic liver disease (ALD). ALD is a global chronic liver disease with potential for hepatocellular carcinoma progression. METHODS: The main active ingredients and targets of garlic were identified through screening the TCMSP, TCM-ID, and ETCM databases. ALD disease targets were sourced from DisGeNET, GeneCards, and DiGSeE databases, and intervention targets for garlic were determined through intersections. Protein interaction networks were constructed using the STRING platform, and GO and KEGG pathway enrichment analyses were performed with R software. The garlic component-disease-target network was established using Cytoscape software. Validation of active ingredients against core targets was conducted through molecular docking simulations using AutoDock Vina software. Expression validation of core targets was carried out using human sequencing data of ALD obtained from the GEO database. RESULTS: Integration of garlic drug targets with ALD disease targets identified 83 target genes. Validation through an alcohol-induced ALD mouse model supported certain network pharmacology findings, suggesting that garlic may impede disease progression by mitigating the inflammatory response and promoting ethanol metabolism. CONCLUSION: This study provides insights into the potential therapeutic mechanisms of garlic in inhibiting ALD development. The identified active ingredients offer promising avenues for further investigation and development of treatments for ALD, emphasizing the importance of botanical remedies in liver disease management.

5.
Nat Commun ; 15(1): 3868, 2024 May 08.
Article in English | MEDLINE | ID: mdl-38719793

ABSTRACT

This study introduces a distributed electrified heating approach that is able to innovate chemical engineering involving endothermic reactions. It enables rapid and uniform heating of gaseous reactants, facilitating efficient conversion and high product selectivity at specific equilibrium. Demonstrated in catalyst-free CH4 pyrolysis, this approach achieves stable production of H2 (530 g h-1 L reactor -1) and carbon nanotube/fibers through 100% conversion of high-throughput CH4 at 1150 °C, surpassing the results obtained from many complex metal catalysts and high-temperature technologies. Additionally, in catalytic CH4 dry reforming, the distributed electrified heating using metallic monolith with unmodified Ni/MgO catalyst washcoat showcased excellent CH4 and CO2 conversion rates, and syngas production capacity. This innovative heating approach eliminates the need for elongated reactor tubes and external furnaces, promising an energy-concentrated and ultra-compact reactor design significantly smaller than traditional industrial systems, marking a significant advance towards more sustainable and efficient chemical engineering society.

6.
Front Immunol ; 15: 1371611, 2024.
Article in English | MEDLINE | ID: mdl-38571940

ABSTRACT

The gasdermin protein family and its homologs in microorganisms have gained significant attention due to their roles in programmed cell death, immune defense, and microbial infection. This review summarizes the current research status of gasdermin proteins, their structural features, and functional roles in fungi, bacteria, and viruses. The review presents evolutionary parallels between mammalian and microbial defense systems, highlighting the conserved role of gasdermin proteins in regulating cell death processes and immunity. Additionally, the structural and functional characteristics of gasdermin homologs in microorganisms are summarized, shedding light on their potential as targets for therapeutic interventions. Future research directions in this field are also discussed to provide a roadmap for further investigation.


Subject(s)
Gasdermins , Animals , Intracellular Signaling Peptides and Proteins/metabolism , Mammals/metabolism , Phosphate-Binding Proteins
7.
Front Microbiol ; 15: 1356415, 2024.
Article in English | MEDLINE | ID: mdl-38435698

ABSTRACT

Tunneling nanotubes (TNTs) are actin-rich intercellular conduits that mediate distant cell-to-cell communication and enable the transfer of various cargos, including proteins, organelles, and virions. They play vital roles in both physiological and pathological processes. In this review, we focus on TNTs in different types of viruses, including retroviruses such as HIV, HTLV, influenza A, herpesvirus, paramyxovirus, alphavirus and SARS-CoV-2. We summarize the viral proteins responsible for inducing TNT formation and explore how these virus-induced TNTs facilitate intercellular communication, thereby promoting viral spread. Furthermore, we highlight other virus infections that can induce TNT-like structures, facilitating the dissemination of viruses. Moreover, TNTs promote intercellular spread of certain viruses even in the presence of neutralizing antibodies and antiviral drugs, posing significant challenges in combating viral infections. Understanding the mechanisms underlying viral spread via TNTs provides valuable insights into potential drug targets and contributes to the development of effective therapies for viral infections.

8.
PLoS One ; 19(1): e0295346, 2024.
Article in English | MEDLINE | ID: mdl-38181024

ABSTRACT

The cell division cycle associated (CDCA) genes regulate the cell cycle; however, their relationship with prognosis in glioma has been poorly reported in the literature. The Cancer Genome Atlas (TCGA) was utilized to probe the CDCA family in relation to the adverse clinical features of glioma. Glioma single-cell atlas reveals specific expression of CDCA3, 4, 5, 8 in malignant cells and CDCA7 in neural progenitor cells (NPC)-like malignant cells. Glioma data from TCGA, the China Glioma Genome Atlas Project (CGGA) and the gene expression omnibus (GEO) database all demonstrated that CDCA2, 3, 4, 5, 7 and 8 are prognostic markers for glioma. Further analysis identified CDCA2, 5 and 8 as independent prognostic factors for glioma. Lasso regression-based risk models for CDCA families demonstrated that high-risk patients were characterized by high tumor mutational burden (TMB), low levels of microsatellite instability (MSI), and low tumor immune dysfunction and rejection (TIDE) scores. These pointed to immunotherapy for glioma as a potentially viable treatment option Further CDCA clustering suggested that the high CDCA subtype exhibited a high macrophage phenotype and was associated with a higher antigen presentation capacity and high levels of immune escape. In addition, hsa-mir-15b-5p was predicted to be common regulator of CDCA3 and CDCA4, which was validated in U87 and U251 cells. Importantly, we found that CDCAs may indicate response to drug treatment, especially rapamycin, in glioma. In summary, our results suggest that CDCAs have potential applications in clinical diagnosis and as drug sensitivity markers in glioma.


Subject(s)
Glioma , Humans , Prognosis , Glioma/drug therapy , Glioma/genetics , Immunotherapy , Computational Biology , Biomarkers , Cell Cycle Proteins/genetics , Nuclear Proteins
9.
Commun Chem ; 6(1): 273, 2023 Dec 13.
Article in English | MEDLINE | ID: mdl-38087001

ABSTRACT

Feedstock properties play a crucial role in thermal conversion processes, where understanding the influence of these properties on treatment performance is essential for optimizing both feedstock selection and the overall process. In this study, a series of van Krevelen diagrams were generated to illustrate the impact of H/C and O/C ratios of feedstock on the products obtained from six commonly used thermal conversion techniques: torrefaction, hydrothermal carbonization, hydrothermal liquefaction, hydrothermal gasification, pyrolysis, and gasification. Machine learning methods were employed, utilizing data, methods, and results from corresponding studies in this field. Furthermore, the reliability of the constructed van Krevelen diagrams was analyzed to assess their dependability. The van Krevelen diagrams developed in this work systematically provide visual representations of the relationships between feedstock and products in thermal conversion processes, thereby aiding in optimizing the selection of feedstock and the choice of thermal conversion technique.

10.
J Am Chem Soc ; 145(41): 22836-22844, 2023 Oct 18.
Article in English | MEDLINE | ID: mdl-37794780

ABSTRACT

Mixed plastic waste treatment has long been a significant challenge due to complex composition and sorting costs. In this study, we have achieved a breakthrough in converting mixed plastic wastes into a single chemical product using our innovative single-atom catalysts for the first time. The single-atom Ru catalyst can convert ∼90% of real mixed plastic wastes into methane products (selectivity >99%). The unique electronic structure of Ru sites regulates the adsorption energy of mixed plastic intermediates, leading to rapid decomposition of mixed plastics and superior cycle stability compared to traditional nanocatalysts. The global warming potential of the entire process was evaluated. Our proposed carbon-reducing process utilizing single-atom catalysts launches a new era of mixed plastic waste valorization.

11.
Proc Natl Acad Sci U S A ; 120(35): e2307625120, 2023 08 29.
Article in English | MEDLINE | ID: mdl-37603751

ABSTRACT

Trehalose plays a crucial role in the survival and virulence of the deadly human pathogen Mycobacterium tuberculosis (Mtb). The type I ATP-binding cassette (ABC) transporter LpqY-SugABC is the sole pathway for trehalose to enter Mtb. The substrate-binding protein, LpqY, which forms a stable complex with the translocator SugABC, recognizes and captures trehalose and its analogues in the periplasmic space, but the precise molecular mechanism for this process is still not well understood. This study reports a 3.02-Å cryoelectron microscopy structure of trehalose-bound Mtb LpqY-SugABC in the pretranslocation state, a crystal structure of Mtb LpqY in a closed form with trehalose bound and five crystal structures of Mtb LpqY in complex with different trehalose analogues. These structures, accompanied by substrate-stimulated ATPase activity data, reveal how LpqY recognizes and binds trehalose and its analogues, and highlight the flexibility in the substrate binding pocket of LpqY. These data provide critical insights into the design of trehalose analogues that could serve as potential molecular probe tools or as anti-TB drugs.


Subject(s)
Mycobacterium tuberculosis , Humans , Cryoelectron Microscopy , Trehalose , ATP-Binding Cassette Transporters , Molecular Probes
12.
Front Genet ; 13: 956632, 2022.
Article in English | MEDLINE | ID: mdl-36186425

ABSTRACT

Guanylate binding protein 2 (GBP2) is a member of the guanine binding protein family, and its relationship with prognostic outcomes and tumor immune microenvironments in glioma remains elusive. We found GBP2 were increased in glioma tissues at both mRNA and protein levels. Kaplan-Meier curves revealed that high GBP2 expression was linked with worse survival of glioma patients, and multivariate Cox regression analysis indicated that high GBP2 expression was an independent prognostic factor for glioma. Combined analysis in immune database revealed that the expression of GBP2 was significantly related to the level of immune infiltration and immunomodulators. Single-cell analysis illustrated the high expression of GBP2 in malignant glioma cells showed the high antigen presentation capability, which were confirmed by real-time polymerase chain reaction (qRT-PCR) data. Additionally, the hsa-mir-26b-5p and hsa-mir-335-5p were predicted as GBP2 regulators and were validated in U87 and U251 cells. Our results first decipher immune-related characteristics and noncoding regulators of GBP2 in glioma, which may provide insights into associated immunotherapies and prognostic predictor.

13.
Front Neurol ; 13: 869880, 2022.
Article in English | MEDLINE | ID: mdl-35937054

ABSTRACT

Intracranial aneurysm is a severe cerebral disorder involving complicated risk factors and endovascular coiling is a common therapeutic selection for intracranial aneurysm. The recurrence is a clinical challenge in intracranial aneurysms after coil embolization. With this study, we provided a meta-analysis of the risk factors for the recurrence of intracranial aneurysm after coil embolization. Nine studies were included with a total of 1,270 studies that were retrieved from the database. The sample size of patients with intracranial aneurysms ranged from 241 to 3,530, and a total of 9,532 patients were included in the present meta-analysis. The intracranial aneurysms that occurred in middle cerebral artery (MCA) (OR = 1.09, 95% CI: 1.03-1.16, P = 0.0045) and posterior circulation (OR = 2.01, 95% CI: 1.55-2.60, P = 0.000) presented the significantly higher risk of recurrence after coil embolization. Meanwhile, intracranial aneurysms of size > 7 mm (OR = 5.38, 95%CI: 3.76-7.70, P = 0.000) had a significantly higher risk of recurrence after coil embolization. Moreover, ruptured aneurysm (OR = 2.86, 95% CI: 2.02-4.04, P = 0.000) and subarachnoid hemorrhage (SAH) (OR = 1.57, 95% CI: 1.20-2.06, P = 0.001) was positively correlated with the risk of recurrence after coil embolization. In conclusion, this meta-analysis identified the characteristics of intracranial aneurysms with MCA, posterior circulation, size > 7 mm, ruptured aneurysm, and SAH as the risk factors of recurrence after coil embolization for intracranial aneurysms.

14.
Water Res ; 222: 118875, 2022 Aug 15.
Article in English | MEDLINE | ID: mdl-35870392

ABSTRACT

The blooming of beach-cast seaweed has caused environmental degradation in some coastal regions. Therefore, a proper treating and utilizing method of beach-cast seaweed is demanded. This study investigated the potential of producing power or biofuel from pyrolysis of beach-cast seaweed and the effect of the ash-washing process. First, the raw and washed beach-cast seaweeds (RS and WS) were prepared. Thereafter, thermogravimetric analysis (TG), bench-scale pyrolysis experiment, process simulation, and life cycle assessment (LCA) were conducted. The TG results showed that the activation energies of thermal decomposition of the main organic contents of RS and WS were 44.23 and 58.45 kJ/mol, respectively. Three peak temperatures of 400, 500, and 600 °C were used in the bench-scale pyrolysis experiments of WS. The 600 °C case yielded the most desirable gas and liquid products. The bench-scale pyrolysis experiment of RS was conducted at 600 °C as well. Also, an LCA was conducted based on the simulation result of 600 °C pyrolysis of WS. The further process simulation and LCA results show that compare to producing liquid biofuel and syngas, a process designed for electricity production is most favored. It was estimated that treating 1 ton of dry WS can result in a negative cumulative energy demand of -2.98 GJ and carbon emissions of -790.89 kg CO2 equivalence.


Subject(s)
Pyrolysis , Seaweed , Animals , Biofuels , Life Cycle Stages , Temperature , Vegetables
15.
Cell Death Discov ; 8(1): 208, 2022 Apr 18.
Article in English | MEDLINE | ID: mdl-35436989

ABSTRACT

Identifying the mechanism of glioma progression is critical for diagnosis and treatment. Although studies have shown that guanylate-binding protein 2(GBP2) has critical roles in various cancers, its function in glioma is unclear. In this work, we demonstrate that GBP2 has high expression levels in glioma tissues. In glioma cells, depletion of GBP2 impairs proliferation and migration, whereas overexpression of GBP2 enhances proliferation and migration. Regarding the mechanism, we clarify that epidermal growth factor receptor (EGFR) signaling is regulated by GBP2, and also demonstrate that GBP2 interacts directly with kinesin family member 22(KIF22) and regulates glioma progression through KIF22/EGFR signaling in vitro and in vivo. Therefore, our study provides new insight into glioma progression and paves the way for advances in glioma treatment.

16.
Mol Med Rep ; 25(3)2022 03.
Article in English | MEDLINE | ID: mdl-35059733

ABSTRACT

Glioblastoma is a common central nervous system tumor and despite considerable advancements in treatment patient prognosis remains poor. Angiogenesis is a significant prognostic factor in glioblastoma, anti­angiogenic treatments represent a promising therapeutic approach. Vascular endothelial growth factor A (VEGFA) is a predominant regulator of angiogenesis and mounting evidence suggests that the Wnt signaling pathway serves a significant role in tumor angiogenesis. As a positive regulator of the Wnt/ß­catenin signaling pathway, frequently rearranged in advanced T­cell lymphomas­1 (FRAT1) is highly expressed in human glioblastoma and is significantly associated with glioblastoma growth, invasion and migration, as well as poor patient prognosis. Bioinformatics analysis demonstrated that both VEGFA and FRAT1 were highly expressed in most tumor tissues and associated with prognosis. However, whether and how FRAT1 is involved in angiogenesis remains to be elucidated. In the present study, the relationship between FRAT1 and VEGFA in angiogenesis was investigated using the human glioblastoma U251 cell line. Small interfering RNAs (siRNAs) were used to silence FRAT1 expression in U251 cells, and the mRNA and protein expression levels of VEGFA, as well as the concentration of VEGFA in U251 cell supernatants, were determined using reverse transcription­quantitative PCR, western blotting and ELISA. A tube formation assay was conducted to assess angiogenesis. The results demonstrated that siRNA knockdown significantly decreased the protein expression levels of FRAT1 in U251 cells and markedly decreased the mRNA and protein expression levels of VEGFA. Furthermore, the concentration of VEGFA in the cell supernatant was significantly reduced and angiogenesis was suppressed. These results suggested that FRAT1 may promote VEGFA secretion and angiogenesis in human glioblastoma cells via the Wnt/ß­catenin signaling pathway, supporting the potential use of FRAT1 as a promising therapeutic target in human glioblastoma.


Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Brain Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Glioblastoma/genetics , Neovascularization, Pathologic/genetics , Proto-Oncogene Proteins/genetics , Vascular Endothelial Growth Factor A/genetics , Adaptor Proteins, Signal Transducing/metabolism , Blotting, Western , Brain Neoplasms/blood supply , Brain Neoplasms/metabolism , Cell Line, Tumor , Cells, Cultured , Female , Glioblastoma/blood supply , Glioblastoma/metabolism , Humans , Male , Middle Aged , Neovascularization, Pathologic/metabolism , Prognosis , Proto-Oncogene Proteins/metabolism , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Vascular Endothelial Growth Factor A/metabolism , Wnt Signaling Pathway/genetics , beta Catenin/genetics , beta Catenin/metabolism
17.
Chemosphere ; 271: 129561, 2021 May.
Article in English | MEDLINE | ID: mdl-33453478

ABSTRACT

A series of magnetic bio-activated carbon (MBAC) has been produced from lignin and ferrous salts following to the process including impregnation, carbonization, and steam activation. The influence of the impregnation methods and the steam flow rate on the quality and the maximum phosphorus adsorption capacity of the produced MBACs has been investigated. The phosphorus adsorption performance in real domestic wastewater of the MBAC with the highest maximum phosphorus adsorption capacity has been investigated. The results show that all of the produced MBACs have a relatively rich porous structure, and all surface iron species exist as magnetite (Fe3O4). Compared with the MBACs that are produced via the dry impregnation method using a lower steam flow rate, the MBACs that are produced via the wet impregnation method using a higher steam flow rate are believed to have a higher iron content and better iron species dispersion. The highest maximum phosphorus adsorption capacity of all the produced MBACs is estimated to be as high as 69.80 mg-P/g according to the best-fitting Langmuir model. The MBAC that shows the highest maximum phosphorus adsorption capacity could also remove 84.65% and 96.97% of the total phosphorus from the filtered raw domestic wastewater (FRDW) and treated domestic wastewater (TDW), respectively, which indicates a good potential for using MBACs for domestic wastewater treatment.


Subject(s)
Charcoal , Wastewater , Adsorption , Magnetic Phenomena , Phosphorus
18.
Oncol Lett ; 20(1): 248-256, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32565951

ABSTRACT

Aberrant expression of frequently rearranged in advanced T-cell lymphomas 1 (FRAT1) contributes to poor prognosis in a number of carcinomas. However, its role in glioma remains controversial. In the present study, gene expression profiling was performed using Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO) functional enrichment and ingenuity pathway analysis (IPA) to evaluate the differential expression of genes and proteins in FRAT1 knockdown U251 glioma cells in comparison with the control. Western blot analysis was conducted to assess the expression levels of FRAT1 and STAT1. A total of 895 downregulated genes were identified in FRAT1-silenced U251 cells. The most enriched processes determined by GO and KEGG analysis of the 895 differentially expressed genes were associated with proliferation, migration and invasion. According to IPA, significant canonical pathways, including the interferon, hepatic fibrosis and Wnt/ß-catenin signaling pathways, were identified to be the major enriched pathways. The elevated expression of STAT1 in U251 cells was validated. These results highlighted the regulatory role of FRAT1 in glioma cells with upregulated STAT1 expression.

19.
Dev Comp Immunol ; 36(1): 191-8, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21801746

ABSTRACT

B-cell activating factor of the TNF family (BAFF) induces B cell survival, proliferation, immunoglobulin secretion and has a role in enhancing immune responses. In the present study, we amplified the cDNA of goat (Capra hircus) BAFF (designated gBAFF) from spleen by reverse transcription-PCR (RT-PCR). The open reading frame (ORF) of gBAFF covers 843 bp encoding 280 amino acids, with a 152-aa mature peptide. Sequence comparison indicated that the amino acid of gBAFF possessed the TNF signature, a transmembrane domain, a putative furin protease cleavage site and three cysteine residues. The predicted three-dimensional (3D) structure of the soluble part of gBAFF (gsBAFF) analyzed by "comparative protein modelling" revealed that it was very similar to its counterparts. Real-time quantitative PCR (qPCR) analysis indicated that gBAFF mRNA was predominantly expressed in goat lymphoid tissue spleen. Recombinant gsBAFF was fused with a small ubiquitin-related modifier (SUMO) gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). The resulting fused protein SUMO-gsBAFF was efficiently expressed and purified using metal chelate affinity chromatography (Ni-NTA), then confirmed by SDS-PAGE and Western blotting analysis. In vitro, the MTT assays and flow cytometric analysis indicated that SUMO-gsBAFF as well as gsBAFF could promote the survival/proliferation of goat splenic B cells or mouse splenic B cells. Therefore, BAFF may be a potential immunologic factor for enhancing immunological efficacy in goat.


Subject(s)
B-Cell Activating Factor/genetics , B-Lymphocytes/metabolism , Goats , Animals , B-Cell Activating Factor/immunology , B-Cell Activating Factor/metabolism , B-Lymphocytes/immunology , B-Lymphocytes/pathology , Base Sequence , Cell Proliferation , Cell Separation , Cloning, Molecular , Computational Biology , Flow Cytometry , Gene Expression Profiling , Lymphocyte Activation , Male , Molecular Sequence Data , Protein Conformation , Sequence Alignment , Transgenes/genetics
20.
Vet Immunol Immunopathol ; 142(1-2): 133-9, 2011 Jul 15.
Article in English | MEDLINE | ID: mdl-21600667

ABSTRACT

B cell activating factor (BAFF), belonging to the tumor necrosis factor (TNF) family, plays an important role in B cell survival, proliferation and differentiation. In the present study, we amplified the cDNA of dog (Canis familiaris) BAFF (designated doBAFF) from spleen by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) strategies. The open reading frame (ORF) of doBAFF covers 879 bp encoding 292 amino acids, with a 152-aa mature peptide. The soluble mature part of doBAFF (dosBAFF) shares 91.5%, 72.1%, 96.7%, 94.0% and 91.5% identity with the human, mouse, cattle, pig and rabbit counterparts, respectively. The predicted three-dimensional (3D) structural analysis of dosBAFF analyzed by "comparative protein modeling" revealed that it was very similar to its human counterpart. Real-time quantitative PCR analysis revealed that doBAFF was mainly expressed in spleen. Two fusion proteins SUMO-dosBAFF and GFP/dosBAFF were efficiently expressed in Escherichia coli BL21 (DE3) and purified using metal chelate affinity chromatography (Ni-NTA). Laser scanning confocal microscopy analysis showed that GFP/dosBAFF could bind to the mouse splenic B-cell. In vitro, SUMO-dosBAFF and GFP/dosBAFF were able to promote the survival/proliferation of dog lymphocytes or mouse splenic B cells with/without anti-IgM. Therefore, BAFF may be a potential immunologic factor for enhancing immunological efficacy in dog.


Subject(s)
B-Cell Activating Factor/genetics , Amino Acid Sequence , Animals , B-Cell Activating Factor/metabolism , B-Cell Activating Factor/physiology , Base Sequence , Blotting, Western/veterinary , Cloning, Molecular , Dogs , Microscopy, Confocal/veterinary , Molecular Sequence Data , Open Reading Frames/genetics , Protein Structure, Tertiary , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Spleen/metabolism
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