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Pericytes are located in the stromal membrane of the capillary outer wall and contain endothelial cells (ECs). They are pivotal in regulating blood flow, enhancing vascular stability, and maintaining the integrity of the blood-retina barrier (BRB)/blood-brain barrier (BBB). The pluripotency of pericytes allows them to differentiate into various cell types, highlighting their significance in vascular disease pathogenesis, as demonstrated by previous studies. This potential enables pericytes to be a potential biomarker for the diagnosis and a target for treatment of vascular disorders. The retina, an essential part of the eyeball, is an extension of cerebral tissue with a transparent refractive medium. It offers a unique window for assessing systemic microvascular lesions. Routine fundus examination is necessary for patients with diabetes and hypertension. Manifestations, such as retinal artery tortuosity, dilation, stenosis, and abnormal arteriovenous anastomosis, serve as typical hallmarks of retinal vasculopathy. Therefore, studies of ocular vascular diseases significantly facilitate the exploration of systemic vascular diseases.
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BACKGROUND: Pathological neovascularization plays a pivotal role in the onset and progression of tumors and neovascular eye diseases. Despite notable advancements in the development of anti-angiogenic medications that target vascular endothelial growth factor (VEGF) and its receptors (VEGFRs), the occurrence of adverse reactions and drug resistance has somewhat impeded the widespread application of these drugs. Therefore, additional investigations are warranted to explore alternative therapeutic targets. In recent years, owing to the swift advancement of high-throughput sequencing technology, pan-cancer analysis and single-cell sequencing analysis have emerged as pivotal methodologies and focal areas within the domain of omics research, which is of great significance for us to find potential targets related to the regulation of pathological neovascularization. METHODS: Pan-cancer analysis and scRNA-seq data analysis were employed to forecast the association between Actin filament-associated protein 1 like 1 (AFAP1L1) and the development of tumors and endothelial cells. Tumor xenograft model and ocular pathological neovascularization model were constructed as well as Isolectin B4 (IsoB4) staining and immunofluorescence staining were used to assess the effects of AFAP1L1 on the progression of neoplasms and neovascular eye diseases in vivo. Transwell assay, wound scratch assay, tube forming assay, three-dimensional germination assay, and rhodamine-phalloidin staining were used to evaluate the impact of AFAP1L1 on human umbilical vein endothelial cells (HUVECs) function in vitro; Dual luciferase reporting, qRT-PCR and western blot were used to investigate the upstream and downstream mechanisms of pathological neovascularization mediated by AFAP1L1. RESULTS: Our investigation revealed that AFAP1L1 plays a crucial role in promoting the development of various tumors and demonstrates a strong correlation with endothelial cells. Targeted suppression of AFAP1L1 specifically in endothelial cells in vivo proves effective in inhibiting tumor formation and ocular pathological neovascularization. Mechanistically, AFAP1L1 functions as a hypoxia-related regulatory protein that can be activated by HIF-1α. In vitro experiments demonstrated that reducing AFAP1L1 levels can reverse hypoxia-induced excessive angiogenic capacity in HUVECs. The principal mechanism of angiogenesis inhibition entails the regulation of tip cell behavior through the YAP-DLL4-NOTCH axis. CONCLUSION: In conclusion, AFAP1L1, a newly identified hypoxia-related regulatory protein, can be activated by HIF-1α. Inhibiting AFAP1L1 results in the inhibition of angiogenesis by suppressing the germination of endothelial tip cells through the YAP-DLL4-NOTCH axis. This presents a promising therapeutic target to halt the progression of tumors and neovascular eye disease.
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Proteínas Adaptadoras Transductoras de Señales , Células Endoteliales , Neovascularización Patológica , Humanos , Inhibidores de la Angiogénesis , Proteínas de Unión al Calcio , Transducción de Señal , Factor A de Crecimiento Endotelial Vascular , AnimalesRESUMEN
INTRODUCTION: The existing selection of driving distraction recognition methods is based on a specific research perspective and does not provide comprehensive information on the entire field of view. METHOD: We conducted a systematic review of previous studies, aiming to come up with appropriate research methods to identify the driver's distraction state. First, this article selects four sets of search keywords related to driving distraction discrimination from five databases (Web of Science, ScienceDirect, Springer Link, IEEE, and TRID) and identifies 1,620 peer-reviewed documents from 2000 to 2020; these 1,620 documents underwent bibliographic analysis and co-occurrence network analysis. The co-occurrence coupling relationship is analyzed from the aspects of time, country, publication, author and keywords. Second, 37 papers published were screened, and the driving distraction recognition methods proposed by these 37 papers were summarized and analyzed. RESULTS: The results show that this field has been prevalent since 2013; countries such as the United States, Britain, Germany, Australia, China, and Canada are in the forefront of research in this field, and the cooperation between related countries is relatively close. The cooperation between authors is characterized by aggregation, and the mobile phone as the main keyword is almost connected to other keyword nodes; the recognition model of deep learning algorithm based on video surveillance data sources has become the mainstream hot spot distraction recognition method. The recognition model of machine learning algorithm based on vehicle dynamics data, driver physiology, and eye movement data sources has specific advantages and disadvantages. PRACTICAL APPLICATIONS: The results can help people to understand the current situation of driving distraction comprehensively and systematically, provide better theoretical support for researchers to choose the subsequent driving distraction recognition model, and provide research direction for driving distraction recognition in the future.
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Conducción de Automóvil , Teléfono Celular , Bibliometría , Bases de Datos Factuales , Humanos , Aprendizaje Automático , Estados UnidosRESUMEN
Background: The metagenomics next-generation sequencing (mNGS) is a promising technique for pathogens diagnosis. However, whether the application of mNGS in critically ill patients with pneumonia could cause anti-infection treatment adjustment and thereby affect the prognosis of these patients has not been explored. Methods: We retrospectively collected the clinical data of patients diagnosed with pulmonary infection in the ICU of the Affiliated Hospital of Qingdao University from January 2018 to January 2021. These patients with pneumonia were divided into mNGS group and no-mNGS group by whether being performed NGS or not. The clinical data, including demographics, illness history, APACHE II score, length of mechanical ventilation, length of stay in the hospital, length of stay in ICU and outcome, were collected. In addition, the data of pathogens and anti-infection treatment before and after NGS were also collected. Propensity score matching was performed to evaluate the mortality and deterioration rate between NGS group and non-NGS group. Results: A total of 641 patients diagnosed with pneumonia were screened, and 94 patients were excluded based on exclusion criteria. Finally, 547 patients were enrolled, including 160 patients being performed NGS. Among these 160 patients, 142 cases had NGS-positive results. In addition, new pathogens were detected in 132 specimens by NGS, which included 82 cases with virus, 18 cases with fungus, 17 cases with bacteria, 14 cases with mycoplasma, and 1 case with mycobacterium tuberculosis. Anti-infection treatments were adjusted in some patients who performed NGS, including 48 anti-bacterial treatments, 20 antifungal treatments and 20 antiviral treatments. There were no significant differences in the mortality and deterioration rate between NGS and non-NGS group, but it exhibited a trend that the mortality and deterioration rate of NGS group was lower than non-NGS group after the propensity score matching analysis (15.8% vs 24.3%, P=0.173; 25.6% vs 37.8%, P=0.093). Conclusion: NGS could affect the anti-infection treatments and had a trend of reducing the mortality and deterioration rate of critically ill patients with pneumonia.
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Mycobacterium tuberculosis , Neumonía , Humanos , Enfermedad Crítica , Estudios Retrospectivos , Neumonía/diagnóstico , Neumonía/tratamiento farmacológico , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Metagenómica/métodos , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: The vitreous body is an important part of the ocular body fluid. A foldable capsular vitreous body (FCVB) is designed to treat chronic adverse complications in severe ocular trauma and silicone oil-dependent eyes. This study is aimed at investigating a method for implanting an FCVB, its postoperative efficacy, and clinical value. METHODS: A retrospective analysis was performed on data from 18 patients who underwent vitrectomy and FCVB implantation for severe ocular trauma and silicone oil-dependent eyes between March 2019 and May 2020. All treated eyes underwent clinical examinations involving the best-corrected visual acuity, intraocular pressure, FCVB position, anterior segment photography, and wide-angle fundus photography regularly after surgery. RESULTS: Eighteen eyes from 18 patients were enrolled in this study. A total of 2.00-4.20 (3.46 ± 0.78) ml of silicone oil were injected into the FCVB during surgery. The patients were followed up at 1, 2, and 4 weeks and 3, 6, and 12 months after surgery. Twelve months after surgery, visual acuity improved in 7 (38.89%) eyes. In contrast, 10 (55.56%) eyes showed no obvious improvement, and 1 (5.56%) eye had decreased vision. Intraocular pressure at 12 months was 10.13 ± 3.52 mmHg, which was comparable to that before the surgery (t = 0.38, P = 0.71). The anterior chamber depth examined by slit lamp was 2.00-3.00 cornea thickness (CT) in 7 eyes, 1.00-2.00 CT in 2 eyes, and <1.00 CT in one eye. The anterior chamber disappeared in eight eyes. There were eight eyes with clear cornea, four eyes with localized opacity, and two eyes with obvious gray-white opacity. There was no case of severe FCVB deflection, rupture, or exposure during the observation period. CONCLUSION: FCVB implantation is an effective and safe treatment for eyes with severe ocular trauma and silicone oil-dependent eyes. It may support retinal reattachment, slow down eyeball atrophy, reduce the risk of chronic adverse complications such as corneal endothelial decompensation, and maintain intraocular pressure and preoperative visual function.
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Aceites de Silicona , Vitrectomía , Cuerpo Vítreo/trasplante , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Epigenetic alterations occur in many physiological and pathological processes. N6-methyladenosine (m6A) modification is the most prevalent modification in eukaryotic mRNAs. However, the role of m6A modification in pathological angiogenesis remains elusive. In this study, we showed that the level of m6A modification was significantly upregulated in endothelial cells and mouse retinas following hypoxic stress, which was caused by increased METTL3 levels. METTL3 silencing or METTL3 overexpression altered endothelial cell viability, proliferation, migration, and tube formation in vitro. METTL3 knockout in vivo decreased avascular area and pathological neovascular tufts in an oxygen-induced retinopathy model and inhibited alkali burn-induced corneal neovascularization. Mechanistically, METTL3 exerted its angiogenic role by regulating Wnt signaling through the m6A modification of target genes (e.g., LRP6 and dishevelled 1 [DVL1]). METTL3 enhanced the translation of LRP6 and DVL1 in an YTH m6A RNA-binding protein 1 (YTHDF1)-dependent manner. Collectively, this study suggests that METTL3-mediated m6A modification is an important hypoxic stress-response mechanism. The targeting of m6A through its writer enzyme METTL3 is a promising strategy for the treatment of angiogenic diseases.
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Adenosina/análogos & derivados , Epigénesis Genética , Regulación de la Expresión Génica , Metiltransferasas/metabolismo , Neovascularización Patológica/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Adenosina/metabolismo , Animales , Biomarcadores , Susceptibilidad a Enfermedades , Silenciador del Gen , Humanos , Hipoxia/complicaciones , Hipoxia/metabolismo , Ratones , Ratones Noqueados , Neovascularización Patológica/metabolismo , Enfermedades de la Retina/etiología , Enfermedades de la Retina/metabolismo , Enfermedades de la Retina/patología , Vía de Señalización WntRESUMEN
The miRNAs and mRNAs are found to play a crucial role in modulating different diseases including stroke, according to the recent evidence. The current study is aimed at assessing the functional role played by miR-188-5p in the regulation of cell apoptosis and viability in OGD-induced human neural cell line HNC. With the help of RT-qPCR, the authors determined miR-188-5p as well as its putative target PTEN among OGD-treated cells in different treatment times. The cell viability was assessed through CCK-8 assay while the cell transfection either upregulated or may have silenced the genes. Both Western Blot as well as RT-qPCR found the proliferation biomarkers such as Ki87 and PCNA in addition to apoptosis biomarkers such as caspase-8 and caspase-3. The luciferase activity was tracked by conducting luciferase assay. The researchers observed an elevation in the expression of miR-188-5p while the PTEN got downregulated in Human Neural Cell line HNC with increase in the time span. The expressions of miR-188-5p and PTEN got increased with increasing OGD treatment time while the Luciferase reassured the binding site. The cell viability was suppressed by the overexpression of miR-188-5p which further inhibited the apoptosis biomarkers too. Meanwhile, it was understood that the results could be reversed to some extent with the inhibition of PTEN. The study findings from in vitro investigations yielded promising results and provided excellent insights about the fundamental molecular mechanisms of miR-188-5p involved in stroke via PTEN. This could be considered as a potential therapeutic axis among stroke patients in the near future.
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Apoptosis/genética , MicroARNs/genética , Fosfohidrolasa PTEN/genética , Accidente Cerebrovascular/genética , Caspasa 8/genética , Línea Celular , Supervivencia Celular/genética , Glucosa/efectos adversos , Humanos , Neuronas/metabolismo , Neuronas/patología , Oxígeno/efectos adversos , Transducción de Señal/genética , Accidente Cerebrovascular/inducido químicamente , Accidente Cerebrovascular/metabolismo , Accidente Cerebrovascular/patologíaRESUMEN
Diabetic retinopathy (DR) is the leading cause of blindness in working-age adults. Vascular pericyte degeneration is the predominant clinical manifestation of DR, yet the mechanism governing pericyte degeneration is poorly understood. Circular RNAs (circRNAs) play important roles in multiple biological processes and disease progression. Here, we investigated the role of circRNA in pericyte biology and diabetes-induced retinal vascular dysfunction. cZNF532 expression was upregulated in pericytes under diabetic stress, in the retinal vessels of a diabetic murine model, and in the vitreous humor of diabetic patients. cZNF532 silencing reduced the viability, proliferation, and differentiation of pericytes and suppressed the recruitment of pericytes toward endothelial cells in vitro. cZNF532 regulated pericyte biology by acting as a miR-29a-3p sponge and inducing increased expression of NG2, LOXL2, and CDK2. Knockdown of cZNF532 or overexpression of miR-29a-3p aggravated streptozotocin-induced retinal pericyte degeneration and vascular dysfunction. By contrast, overexpression of cZNF532 or inhibition of miR-29a-3p ameliorated human diabetic vitreous-induced retinal pericyte degeneration and vascular dysfunction. Collectively, these data identify a circRNA-mediated mechanism that coordinates pericyte biology and vascular homeostasis in DR. Induction of cZNF532 or antagonism of miR-29a-3p is an exploitable therapeutic approach for the treatment of DR.
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Retinopatía Diabética/metabolismo , Pericitos/metabolismo , ARN Circular/metabolismo , Vasos Retinianos/metabolismo , Animales , Línea Celular , Retinopatía Diabética/genética , Retinopatía Diabética/patología , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Humanos , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Pericitos/patología , ARN Circular/genética , Vasos Retinianos/patologíaRESUMEN
The crosstalk between vascular pericytes and endothelial cells (ECs) is critical for microvascular stabilization and remodeling; however, the crosstalk is often disrupted by diabetes, leading to severe and even lethal vascular damage. Circular RNAs are a class of endogenous RNAs that regulate several important physiological and pathological processes. Here we show that diabetes-related stress up-regulates cPWWP2A expression in pericytes but not in ECs. In vitro studies show that cPWWP2A directly regulates pericyte biology but indirectly regulates EC biology via exosomes carrying cPWWP2A. cPWWP2A acts as an endogenous miR-579 sponge to sequester and inhibit miR-579 activity, leading to increased expression of angiopoietin 1, occludin, and SIRT1. In vivo studies show that cPWWP2A overexpression or miR-579 inhibition alleviates diabetes mellitus-induced retinal vascular dysfunction. By contrast, inhibition of cPWWP2A-mediated signaling by silencing cPWWP2A or overexpressing miR-579 aggravates retinal vascular dysfunction. Collectively, this study unveils a mechanism by which pericytes and ECs communicate. Intervention of cPWWP2A or miR-579 expression may offer opportunities for treating diabetic microvascular complications.
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Comunicación Celular , Retinopatía Diabética/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , MicroARNs/biosíntesis , Pericitos/metabolismo , Transducción de Señal , Regulación hacia Arriba , Animales , Retinopatía Diabética/patología , Exosomas/metabolismo , Exosomas/patología , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Masculino , Ratones , MicroARNs/genética , Pericitos/patología , Vasos Retinianos/metabolismo , Vasos Retinianos/patologíaRESUMEN
Proliferative vitreoretinopathy (PVR) is one of the major challenges in retinal surgery, which occurs in the patient with complex retinal surgery or penetrating eye injury. Circular RNAs (circRNAs) have emerged as important regulators in many biological processes and disease development. However, the characterization and function of circRNAs in PVR remains elusive. In this study, we identified 91 dysregulated circRNAs in the epiretinal membranes (ERMs) of PVR patients. We further investigated the expression pattern of circ_0043144. circ_0043144 was significantly up-regulated in the vitreous samples and the corresponding serum samples of the patients with PVR. circ_0043144 expression was significantly down-regulated after PVR operation. In vitro studies revealed that circ_0043144 was involved in the regulation of the proliferation, migration and secretion ability of ARPE-19 cells, which is critical for ERM formation. Collectively, this study indicates that circRNAs are potential regulators of the pathogenesis of PVR. circ_0043144 is a promising prognostic and diagnostic indicator for PVR diseases.
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Perfilación de la Expresión Génica/métodos , ARN/genética , ARN/metabolismo , Vitreorretinopatía Proliferativa/genética , Vitreorretinopatía Proliferativa/metabolismo , Células Cultivadas , Membrana Epirretinal/genética , Membrana Epirretinal/metabolismo , Membrana Epirretinal/patología , Humanos , ARN Circular , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/patología , Vitreorretinopatía Proliferativa/patologíaRESUMEN
Antibody-based therapy is an effective strategy for treating ocular angiogenesis. However, short-acting efficacy and poor treatment compliance usually occurs in clinical practices. Thus, it is required to develop a drug delivery system to improve the bioavailability and decrease the toxicity of anti-angiogenic antibody. Bevacizumab is a recombinant humanized monoclonal antibody against vascular endothelial growth factor (VEGF). In this study, bevacizumab was encapsulated into poly (lactide-co-glycolide) (PLGA) nanoparticles. PLGA encapsulation could prolong the residency of bevacizumab in the vitreous and aqueous humor and produce long-lasting drug concentrations. Bevacizumab-encapsulated PLGA had no significant cytotoxicity and tissue toxicity effect in vitro and in vivo. In vitro studies showed that bevacizumab-encapsulated PLGA was more effective than bevacizumab in inhibiting VEGF-mediated endothelial cell proliferation, migration and tube formation. In vivo studies showed that bevacizumab-encapsulated PLGA enhanced the anti-angiogenic efficiency of bevacizumab for treating corneal neovascularization and retinal neovascularization. Thus, bevacizumab-encapsulated PLGA could increase the bioavailability and decrease the toxicity of bevacizumab during ocular angiogenesis therapy.
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Inhibidores de la Angiogénesis/administración & dosificación , Bevacizumab/administración & dosificación , Neovascularización de la Córnea/tratamiento farmacológico , Neovascularización Retiniana/tratamiento farmacológico , Inhibidores de la Angiogénesis/farmacología , Inhibidores de la Angiogénesis/toxicidad , Animales , Humor Acuoso/metabolismo , Bevacizumab/farmacología , Bevacizumab/toxicidad , Disponibilidad Biológica , Neovascularización de la Córnea/patología , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Nanopartículas , Tamaño de la Partícula , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Neovascularización Retiniana/patología , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Cuerpo Vítreo/metabolismoRESUMEN
BACKGROUND/AIMS: Pterygium is a common ocular surface disease with an unknown etiology and threatens vision as it invades into the cornea. Circular RNAs (circRNAs) are a novel class of RNA transcripts that participate in several physiological and pathological processes. However, the role of circRNAs in pathogenesis of pterygium remains largely unknown. METHODS: Genome-wide circRNA expression profiling was performed to identify pterygium -related circRNAs. GO analysis, pathway analysis, and miRNA response elements analysis was performed to predict the function of differentially expressed circRNAs in pterygium. MTT assays, Ki67 staining, Transwell assay, Hoechst 33342 staining, and Calcein-AM/PI staining were performed to determine the effect of circRNA silencing on pterygium fibroblast and epithelial cell function. RESULTS: Approximately 669 circRNAs were identified to be abnormally expressed in pterygium tissues. GO analysis demonstrated that the host genes of differentially expressed circRNAs were targeted to extracellular matrix organization (ontology: biological process), cytoplasm (ontology: cellular component), and protein binding (ontology: molecular function). Pathway analysis showed that dysregulated circRNAs-mediated regulatory networks were mostly enriched in focal adhesion signaling pathway. Notably, circ_0085020 (circ-LAPTM4B) was shown as a potential biomarker for pterygium. circ_0085020 (circ-LAPTM4B) silencing affected the viability, proliferation, migration, and apoptosis of pterygium fibroblast and epithelial cells in vitro. CONCLUSIONS: This study provides evidence that circRNAs are involved in the pathogenesis of pterygium and might constitute promising targets for the therapeutic intervention of pterygium.
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Células Epiteliales , Fibroblastos , Estudio de Asociación del Genoma Completo , Pterigion , ARN , Biomarcadores/metabolismo , Células Epiteliales/metabolismo , Células Epiteliales/patología , Fibroblastos/metabolismo , Fibroblastos/patología , Humanos , Pterigion/genética , Pterigion/metabolismo , Pterigion/patología , ARN/biosíntesis , ARN/genéticaRESUMEN
VEGF-induced neovascularization plays a pivotal role in corneal neovascularization (CoNV). The current study investigated the potential effect of ginsenoside Rh2 (GRh2) on neovascularization. In HUVECs, pretreatment with GRh2 largely attenuated VEGF-induced cell proliferation, migration, and vessel-like tube formation in vitro. At the molecular level, GRh2 disrupted VEGF-induced VEGF receptor 2 (VEGFR2)-Grb-2-associated binder 1 (Gab1) association in HUVECs, causing inactivation of downstream AKT and ERK signaling. Gab1 knockdown (by targeted short hairpin RNA) similarly inhibited HUVEC proliferation and migration. Notably, GRh2 was ineffective against VEGF in Gab1-silenced HUVECs. In a mouse cornea alkali burn model, GRh2 eyedrops inhibited alkali-induced neovascularization and inflammatory cell infiltrations in the cornea. Furthermore, alkali-induced corneal expression of mRNAs/long noncoding RNAs in cornea were largely attenuated by GRh2. Overall, GRh2 inhibits VEGF-induced angiogenic effect via inhibiting VEGFR2-Gab1 signaling in vitro. It also alleviates angiogenic and inflammatory responses in alkali burn-treated mouse corneas.-Zhang, X.-P., Li, K.-R., Yu, Q., Yao, M.-D., Ge, H.-M., Li, X.-M., Jiang, Q., Yao, J., Cao, C. Ginsenoside Rh2 inhibits vascular endothelial growth factor-induced corneal neovascularization.
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Antiinflamatorios/farmacología , Neovascularización de la Córnea/tratamiento farmacológico , Ginsenósidos/farmacología , Proteínas Adaptadoras Transductoras de Señales , Animales , Antiinflamatorios/uso terapéutico , Córnea/efectos de los fármacos , Córnea/metabolismo , Neovascularización de la Córnea/etiología , Neovascularización de la Córnea/metabolismo , Ginsenósidos/uso terapéutico , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Hielo , Sistema de Señalización de MAP Quinasas , Masculino , Ratones Endogámicos ICR , Fosfoproteínas/metabolismo , Factor A de Crecimiento Endotelial Vascular/toxicidad , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Excessive light exposure leads to retinal degeneration and accelerates the progression and severity of several ocular diseases, such as age-related macular degeneration (AMD) and retinitis pigmentosa. Long non-coding RNAs (LncRNAs) have emerged as important regulators of photoreceptor development and ocular diseases. In this study, we investigated the role of lncRNA-MEG3 in light-induced retinal degeneration. MEG3 expression was significantly up-regulated after light insult in vivo and in vitro. MEG3 silencing protected against light-induced retinal degeneration in vivo and light-induced photoreceptor cell apoptosis in vitro. Mechanistically, MEG3 regulated retinal photoreceptor cell function by acting as p53 decoy. MEG3 silencing decreased caspase 3/7 activity, up-regulated anti-apoptotic protein (Bcl-2) expression, and down-regulated pro-apoptotic protein (Bax) expression. Taken together, this study provides a promising method of MEG3 silencing for treating light-induced retinal degeneration.
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Terapia Genética/métodos , ARN Largo no Codificante/genética , Traumatismos Experimentales por Radiación/genética , Traumatismos Experimentales por Radiación/prevención & control , Degeneración Retiniana/genética , Degeneración Retiniana/prevención & control , Animales , Silenciador del Gen , Luz/efectos adversos , Masculino , Ratones , Ratones Endogámicos C57BL , Traumatismos Experimentales por Radiación/patología , Degeneración Retiniana/patología , Resultado del TratamientoRESUMEN
A novel efficient proteolysis approach was developed based on trypsin-immobilized miniature incandescent bulbs and infrared (IR) radiation. Trypsin was covalently immobilized in the chitosan coating on the outer surface of miniature incandescent bulbs with the aid of glutaraldehyde. When an illuminated enzyme-immobilized bulb was immersed in protein solution, the emitted IR radiation could trigger and accelerate heterogeneous protein digestion. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of hemoglobin (HEM), cytochrome c (Cyt-c), lysozyme (LYS), and ovalbumin (OVA) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF-MS with the sequence coverages of 91%, 77%, 80%, and 52% for HEM, Cyt-c, LYS, and OVA (200 ng µL(-1) each), respectively. The suitability of the prepared bulb bioreactors to complex proteins was demonstrated by digesting human serum.
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Enzimas Inmovilizadas/metabolismo , Rayos Infrarrojos , Iluminación , Proteolisis , Tripsina/metabolismo , Reactores Biológicos , Citocromos c/química , Citocromos c/metabolismo , Enzimas Inmovilizadas/química , Hemoglobinas/química , Hemoglobinas/metabolismo , Humanos , Muramidasa/química , Muramidasa/metabolismo , Ovalbúmina/química , Ovalbúmina/metabolismo , Tripsina/químicaRESUMEN
A graphene-cobalt microsphere hybrid was prepared by the chemical reduction of a mixture containing graphene oxide and cobalt chloride. It was found that a little amount of potassium borohydride or sodium borohydride could initiate the hydrazine-reduction of the mixture at a low temperature of 80 °C. The structure of the material was investigated by scanning electron microscopy, transmission electron microscopy, energy dispersive spectroscopy, vibrating sample magnetometery, Fourier transform infrared spectroscopy, Raman spectroscopy, and Brunauer-Emmett-Teller (BET) techniques. The average size of the cobalt microspheres on graphene sheets was measured to be â¼590 nm. Magnetic investigations indicated that the graphene-cobalt microsphere hybrid exhibited ferromagnetic behavior at room temperature. In addition, the magnetic hybrid was successfully employed in the enrichment and identification of low-abundance proteins and peptides in combination with mass spectrometry.
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A partially purified potato polyphenol oxidase (PPO) was immobilized in a cross-linked chitosan-SiO2 gel and used to treat phenol solutions. Under optimized conditions (formaldehyde 20 mg/ml, PPO 4 mg/ml and pH 7.0), the activity of immobilized PPO was 1370 U/g and its Km value for catechol was 12 mM at 25 degrees C. The highest activity of immobilized enzyme was at pH 7.4. Immobilization stabilized the enzyme with 73 and 58% retention of activity after 10 and 20 days, respectively, at 30 degrees C whereas most of the free enzyme was inactive after 7 days. The efficiency of removing phenol (10 mg phenol/l) by the immobilized PPO was 86%, and about 60% removal efficiency was retained after five recycles. The immobilized PPO may thus be a useful for removing phenolic compounds from industrial waste-waters.