Objective: To explore the effect of Ephrin-B2 on angiogenesis in the ischemic penumbra after transient focal cerebral ischemia in rats; To clarify the mechanism of Ephrin-B2 triggering angiogenesis after transient focal cerebral ischemia. Methods:Sprague-Dawley rats were randomly divided into three groups: normal group, ischemic-reperfusion group and Ephrin-B2 treated group,the last two groups were randomly divided into isubgroups of ischemic-reperfusion on the 4th, 7th, 14 th, and 28 th day. Longa methods were used to make the focal middle cerebral artery occlusion model, and the ischemic brain was reperfused 2 hours after occlusion; Ephrin-B2 protein was administered intracerebroventricularly to examine its effect on angiogenesis and behavioral recovery.Microvessel density(MVD) was quantified by counting the number of CD31+and BrdU+ cells. Rats were subjected to neurologic functional tests by modified neurological severity scores(mNSS) before sacrificed. Western-blot and quantitative real-time reverse-transcription Polymerase chain reaction were used to detect the dynamic expression profile of VEGF(Vascular Endothelial Growth Factor) in the penumbra cortex. Double immunoflurence was used to speculate the location and the co-expression of VEGF with blood vessels. Results:Compared to the normal group, mNSS of Ephrin-B2 treated group gradually declined(P 0.05), the MVD of penumbra cortex in Ephrin-B2 treated group increased(P0.05), and VEGF protein and mRNA level of the penumbra cortex in Ephrin-B2 treated group was highly upregulated after reperfusion in all of subgroups(P0.05); Double immunoflurence indicated that VEGF were expressed in blood vessels. Conclusion: Ephrin-B2 may improve post-stroke functional recovery by enhancing VEGF-induced angiogenesis.
Abstract Introduction: Peripheral immune response has been revealed to play a critical role in proliferative vitreoretinopathy (PVR). However, the reliable immune-related factors that are acting as prognostic indicators or therapeutic targets for PVR remain to explore further. Methods: In the current study, we applied whole-transcriptome sequencing to profile peripheral blood mononuclear cells (PBMCs) from PVR patients and also analyzed lncRNA-mRNA interactions in peripheral immune cells to explore the pathways that might mediate immunopathology and resultant retinal damage in PVR. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses and Ingenuity Pathway Analysis (IPA) were employed to classify the function of these differentially expressed genes (DEGs). Results: Compared to the controls, there were 319 genes upregulated, and 191 genes downregulated in PVR patients. GO, and KEGG enrichment analyses as well as IPA showed that these upregulated genes were significantly enriched in immune-related and infection-relate terms. Immune-related gene NFKBIA , CXCL2 , and CXCL8 were detected as hub-genes in the co-expression network, while lncRNAs such as AC007032.1 , AC037198.2 , AL929472.2 , and SLED1 were highly co-expressed with them. lncRNA-mRNA interactions analysis also showed that putative targeted genes of these differentially expressed lncRNAs were also significantly enriched in immune-related or infection-relate pathways . Conclusion: Our study highlights the transformation of immune-related genes/pathways in PVR by comparing controls, and validates several critical genes and lncRNAs, which are serving as potential diagnostic markers for PVR patients.
Dynamic epigenetic alterations accompanying CD4+ T helper cell differentiation have been implicated in multiple autoimmune diseases. The bromodomain and extra-terminal (BET) proteins are epigenetic regulators that recognize and bind to acetylated histones in chromatin and are targets for pharmacological inhibition. In this study we tested a new BET inhibitor under clinical development, OTX015, to interrogate its effects on key CD4+ T cell subsets associated with autoimmunity.Naïve and memory murine and human CD4+ T cells were isolated and differentiated into populations characterized by the expression of interferon (IFN)-γ and interleukin (IL)-17. Cultured cells were then exposed to varying concentrations of OTX015 in vitro, and its impact on cytokine expression was quantified by flow cytometry. In parallel, the expression of the transcription factors TBX21 and RORC was quantified by PCR. A previously studied BET inhibitor JQ1 was used as a pharmacological control.OTX015 suppressed both murine and human CD4+ T cell proliferation. Its impact on cytokine expression varied in murine and human naïve and memory subsets. OTX015 was similarly effective as JQ1 in the suppression of cytokines and T helper cell proliferation. Higher concentrations of OTX015 also had a greater impact on the viability of murine versus human cells. IL-17 and IFN-γ expression was not altered in murine memory CD4+ T cells, whereas in human memory CD4+ T cells, OTX015 inhibited IL-17, but not IFN-γ. Across all human T cell subsets OTX015 suppressed IL-17 more effectively than IFN-γ.Our studies demonstrate that OTX015 has anti-inflammatory effects by suppressing murine and human CD4+ T cell proliferation and subset-dependent proinflammatory cytokine expression, including the selective suppression of IL-17 in human memory CD4+ T cells.
The external knowledge-enhanced task-oriented dialogues system is designed to cover the user requests beyond the predefined data base (DB) and application-specific interface (API). Existing methods mainly focus to improve the retrieval accuracy of the external knowledge but ignore the modeling of the entity-aware dialog intention, which cannot fast locate the document-level external API in the practical scenarios. Thus, this paper has introduced the entity-aware dialogue intention information and proposed a two-stage training method for the intention-guided knowledge document retrieval model, which can effectively enhance the retrieval accuracy of the Bi-encoder retrieval model in the task-based dialogue system. Besides, to further improve the generalization ability of the retrieval model based on the intention-guided, a meta-task based on the domain offset is constructed and make the Bi-encoder retrieval model get the more robust semantic representation through the meta-learning, which can improve the generalization capability of the model in invisible domains. Experimental results are shown to prove the correctness and effectiveness of the proposed strategy.
To investigate the survival and biofilm formation capacity of Candida albicans in starvation and under anaerobic conditions.Candida albicans growth and survival were monitored in vitro for up to 8 months. Fungal suspensions from late exponential, stationary and starvation phases were incubated on human dentine, polystyrene and glass slides. Scanning electron microscopy (SEM) was used to observe the process of biofilm formation. 2,3-bis(2-Methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide inner salt (XTT) reduction assay was performed to quantify the biofilm formation capability, and confocal laser scanning microscopy (CLSM) was used to study and make semi-quantitative comparisons of the ultrastructure of biofilms formed on human dentine. 'XTT bioactivity' and 'COMSTAT results' were analysed by two-way analysis of variance (ANOVA) and one-way ANOVA, respectively.Candida albicans survived for over six months. SEM demonstrated that starving C. albicans produced mature biofilms on different substrata. C. albicans of the same growth phase incubated on human dentine displayed significantly higher biofilm formation capability than on polystyrene or glass slides (P < 0.05). Biofilm formation capability by starving cells was significantly lower than that in exponential or stationary phases (P < 0.05). CLSM revealed that biofilms formed by starvation-phase cells were less complex, had a higher roughness coefficient and surface/volume ratio (P < 0.05).Candida albicans cells can survive and form biofilms in anaerobic and nutrient-limited conditions and may pose a treatment challenge.
Purpose: A growing body of evidence suggests that the microbiome of the ocular surface confers potent immunoregulatory functions and has a key role in the physiologic maintenance of healthy eyes and in the pathogenesis of ocular diseases. Although the microbiome is known to be affected by age and sex, the influence of these factors on ocular surface microbiota in healthy adults remains largely unknown. Methods: Ocular surface microbiome samples were obtained from the inferior bulbar conjunctiva of 48 young and 42 old adults at Zhongshan Ophthalmic Center. Using metagenomic shotgun sequencing, we characterized the sex- and age-differences in conjunctival microbiome profiles of healthy adults. Results: Male and female groups differed only in the β diversity of bacterial communities, while there were significant differences in bacterial composition, metabolic functions, and the abundance of antibiotic resistance genes between young and old adult groups. Conclusions: Our findings suggest that age and sex collectively shape the conjunctival microbiome, and may change the immune homeostasis of the ocular surface through alterations of its commensal microbiome.
Age-related macular degeneration (AMD) is a progressive and irreversible eye disease. The anti-vascular endothelial growth factor (VEGF) therapy has revolutionized the treatment of neovascular AMD. However, the expense for such treatment is quite high.We used a traditional Chinese medicine ZQMT as an alternative therapeutic regimen for AMD. We employed two in vivo animal models mimicking dry and wet AMD respectively to assess the therapeutic efficacy of ZQMT on treating AMD-related retinopathy. AMD-related retinopathy in Crb1rd8 mice was evaluated from week 1 to 8 by fundus photography. Laser-induced choroidal neovascularization (CNV) was evaluated by fluorescein angiography and histopathology.ZQMT increased CX3CR1 expression in murine CD4+ T cells either cultured in vitro or directly isolated from animals treated with ZQMT. We also performed both in vitro and in vivo studies to confirm that ZQMT has no apparent toxic effects. ZQMT alleviated AMD-related retinopathy in both Crb1rd8 and CNV models. Depletion of CCL2 and CX3CR1 in Crb1rd8 mice abolished the efficacy of ZQMT, suggesting that CCL2 and/or CX3CR1 may underlie the mechanisms of ZQMT in treating AMD-related retinopathy in mice.In summary, our study supports the protective roles of a traditional Chinese medicine ZQMT in AMD.
Angiogenesis is tightly controlled by growth factors and cytokines in pathophysiological settings. Interleukin 37 (IL-37) is a newly identified cytokine of the IL-1 family, some members of which are important in inflammation and angiogenesis. However, the function of IL-37 in angiogenesis remains unknown. We aimed to explore the regulatory role of IL-37 in pathological and physiological angiogenesis.We found that IL-37 was expressed and secreted in endothelial cells and upregulated under hypoxic conditions. IL-37 enhanced endothelial cell proliferation, capillary formation, migration, and vessel sprouting from aortic rings with potency comparable with that of vascular endothelial growth factor. IL-37 activates survival signals including extracellular signal-regulated kinase 1/2 and AKT in endothelial cells. IL-37 promoted vessel growth in implanted Matrigel plug in vivo in a dose-dependent manner with potency comparable with that of basic fibroblast growth factor. In the mouse model of retinal vascular development, neonatal mice administrated with IL-37 displayed increased neovascularization. We demonstrated further that IL-37 promoted pathological angiogenesis in the mouse model of oxygen-induced retinopathy.Our findings suggest that IL-37 is a novel and potent proangiogenic cytokine with essential role in pathophy siological settings.
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