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The critical factors regulating stem cell endothelial commitment and renewal remain not well understood. Here, using loss- and gain-of-function assays together with bioinformatic analysis and multiple model systems, we show that PDGFD is an essential factor that switches on endothelial commitment of embryonic stem cells (ESCs). PDGFD genetic deletion or knockdown inhibits ESC differentiation into EC lineage and increases ESC self-renewal, and PDGFD overexpression activates ESC differentiation towards ECs. RNA sequencing reveals a critical requirement of PDGFD for the expression of vascular-differentiation related genes in ESCs. Importantly, PDGFD genetic deletion or knockdown increases ESC self-renewal and decreases blood vessel densities in both embryonic and neonatal mice and in teratomas. Mechanistically, we reveal that PDGFD fulfills this function via the MAPK/ERK pathway. Our findings provide new insight of PDGFD as a novel regulator of ESC fate determination, and suggest therapeutic implications of modulating PDGFD activity in stem cell therapy.
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Células-Tronco Embrionárias , Modelos Biológicos , Animais , Diferenciação Celular/genética , Células-Tronco Embrionárias/metabolismo , Sistema de Sinalização das MAP Quinases , CamundongosRESUMO
BACKGROUND: Diabetic retinopathy (DR) is the common complication of diabetes, accounting for most blindness cases worldwide. MicroRNAs (miRs) are small non-coding RNAs and participate in the pathogenesis and develop-ment of various diseases, including DR. The present study aimed to investigate miR-335-3p and vascular endothelial growth factor (EGFR) roles in DR diagnosis and development. METHODS: A total of 104 healthy volunteers, 96 type 2 diabetes mellitus (T2DM) patients, and 102 DR cases were enrolled in this study. The clinicopathological information of all subjects were collected and analyzed using chisquared test. After collecting plasma from each participant, a ROC assay was conducted to determine the dis-criminative value of miR-335-3p and EGFR in DR diagnosis. The targeted relationship between miR-335-3p and EGFR was examined by dual-luciferase reporter assay and correlation analysis. After exposing APRE-19 cells to different concentrations of high glucose (HG), the DR in vitro cell model was constructed. The expression levels of miR-335-3p and EGFR were detected using RT-qPCR. The effects of miR-335-3p and EGFR on HG-treated APRE-19 cell viability were determined by CCK-8 assay. RESULTS: Clinicopathological information presented that BMI index, fasting blood glucose (FBP), 2h-BG, HbA1c, miR-335-3p, and EGFR levels were strongly associated with DR pathogenesis. MiR-335-3p was significantly decreased while EGFR was increased in DR patients and HG-treated APRE-19 cells. MiR-335-3p and EGFR presented high accuracy, sensitivity, and specificity in differentiating DR from healthy cases and T2DM patients; moreover, miR-335-3p and EGFR could also discriminate proliferative DR (PDR) cases from healthy controls. After confirming that miR-335-3p was negatively correlated with its target EGFR, we found miR-335-3p could increase the viability in HG-treated APRE-19 cells while silencing of EGFR could also reverse the inhibitory effects of HG conditions on APRE-19 cell viability. CONCLUSIONS: We concluded that plasma miR-335-3p and EGFR may be utilized as non-invasive biomarkers for screening DR cases, contributing to DR diagnosis and treatment.
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Diabetes Mellitus Tipo 2 , Retinopatia Diabética , MicroRNAs , Proliferação de Células , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/genética , Retinopatia Diabética/diagnóstico , Retinopatia Diabética/genética , Fatores de Crescimento Endotelial/farmacologia , Receptores ErbB/genética , Humanos , Fator A de Crescimento do Endotélio VascularRESUMO
BACKGROUND: To detect the expression of histone methyltransferase SETDB1 in hepatocellular carcinoma, and to analyze the relationship between SETDB1 expression and tumor size, microvascular invasion, pTNM stage, gender, age, tumor number, tumor differentiation, and other clinicopathological characteristics. METHODS: Immunohistochemical method was used to detect the expression of SETDB1 proteins in liver cancer tissues and adjacent tissues of 100 cases. The qRT-PCR method was used to detect the expression of SETDB1 mRNA in hepatocellular carcinoma and adjacent tissues of 64 cases. RESULTS: The expression of SETDB1 protein and mRNA in hepatocellular carcinoma was higher than that of adjacent normal liver tissue (p < 0.05). High protein expression of SETDB1 was associated with tumor size, MVI presence, and pTNM stage (p < 0.05). Univariate analysis revealed that the tumor size, tumor differentiation, MVI grade, and pTNM stage were correlated with DFS, while tumor size, MVI grade, pTNM stage, and SETDB1 protein expression were correlated with OS. Multivariate analysis showed that the combination of MVI grade and pTNM stage has statistical significance in predicting prognosis, while SETDB1 protein expression was not significant prognosis factor. CONCLUSIONS: SETDB1 has a certain role in HCC progression and may act as a prognostic predictor concerning the survival of HCC patients.
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Carcinoma Hepatocelular , Histona-Lisina N-Metiltransferase , Neoplasias Hepáticas , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Epigênese Genética/genética , Feminino , Histona-Lisina N-Metiltransferase/genética , Histona-Lisina N-Metiltransferase/metabolismo , Humanos , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Transcriptoma/genéticaRESUMO
BACKGROUND Dysregulation of the microRNA (miRNA) network is a typical feature of many cancers. However, the key specific miRNAs involved in uveal melanoma carcinogenesis are largely unknown. MATERIAL AND METHODS RT-qPCR was used to detected miR-652 expression in uveal melanoma tissues and cell lines. miR-652 inhibitor was transfected into uveal melanoma cells to decrease miR-652 expression and determine the biological role of miR-652 by CCK-8 and wound healing assays. Bioinformatic analysis and dual luciferase reporter assay were used to predict and validate the target gene of miR-652. HOXA9 siRNA was transfected into cells to confirm that miR-652 relies on regulation of HOXA9 to regulate cell proliferation and migration. RESULTS RT-qPCR showed that miR-652 was overexpressed in uveal melanoma cell lines (MUM-2B, MEL270) compared with melanocyte cells (ARPE-19). Overexpression of miR-652 was also observed in uveal melanoma compared to paired non-tumor tissues. Downregulation of miR-652 inhibited the cell proliferation ability and migration ability of uveal melanoma cells. Using bioinformatic analysis, HOXA9 was found to be a potential target gene of miR-652. The direct regulation of HOXA9 by miR-652 was experimentally validated in uveal melanoma cells by dual luciferase assay and Western blotting. We also observed that miR-652 promoted HIF-1alpha signaling via repression of HOXA9 in uveal melanoma cells. Silencing of HOXA9 attenuated the miR-652 inhibitor decreased cell growth rate and decreased migration ability in uveal melanoma cells. CONCLUSIONS Our data demonstrate an oncogenic role of miR-652 in uveal melanoma, showing that miR-652 may be a useful biomarker for prediction of prognosis for patients with uveal melanoma.
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Proteínas de Homeodomínio/genética , Melanoma/genética , MicroRNAs/genética , Neoplasias Uveais/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Melanoma/metabolismo , MicroRNAs/metabolismo , Transdução de Sinais , Neoplasias Uveais/metabolismoRESUMO
Cataract, the leading cause of blindness worldwide, is caused by crystallin protein aggregation within the protected lens environment. Phase separation has been implicated as an important mechanism of protein aggregation diseases, such as neurodegeneration. Similarly, cataract has been proposed to be a protein condensation disease in the last century. However, whether crystallin proteins aggregate via a phase separation mechanism and which crystallin protein initiates the aggregation remain unclear. Here, we showed that all types of crystallin-GFP proteins remain soluble under physiological conditions, including protein concentrations, ion strength, and crowding environments. However, in age or disease-induced aberrant conditions, α-crystallin-GFP, including αA- and αB-crystallin-GFP, but not other crystallin-GFP proteins, undergo phase separation in vivo and in vitro. We found that aging-related changes, including higher crystallin concentrations, increased Na+, and decreased K+ concentrations, induced the aggregation of α-crystallin-GFP. Furthermore, H2O2, glucose, and sorbitol, the well-known risk factors for cataract, significantly enhanced the aggregation of αB-crystallin-GFP. Taken together, our results revealed that α-crystallin-GFP forms aggregates via a phase transition process, which may play roles in cataract disease. Opposite to the previously reported function of enhancing the solubility of other crystallin, α-crystallin may be the major aggregated crystallin in the lens of cataract patients.
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Catarata , Cristalinas , Cristalino , Cadeia A de alfa-Cristalina , alfa-Cristalinas , Humanos , alfa-Cristalinas/metabolismo , Cristalinas/genética , Cristalinas/metabolismo , Agregados Proteicos , Peróxido de Hidrogênio/metabolismo , Catarata/metabolismo , Cristalino/metabolismoRESUMO
Non-infectious chronic diseases, especially inflammatory bowel diseases (IBDs), hypertension, and diabetes mellitus, are characterized by a prolonged and multisystemic course, and their incidence increases annually, usually causing serious economic burden and psychological stress for patients. Therefore, these diseases deserve scientific and consistent disease management. In addition, the lack of a comprehensive "early disease clues tracking-personalized treatment system-follow-up" model in hospitals also exacerbates this dilemma. Based on these facts, we propose an individualized prediction management system for IBDs based on chronic diseases, focusing on the established IBDs-related prediction models and summarizing their advantages and disadvantages. We call on researchers to pay attention to the integration of models with clinical practice and the continuous correction of models to achieve truly individualized medical treatment for chronic diseases, thus providing substantial value for the rapid diagnosis and adequate treatment of chronic diseases such as IBDs, which follow the "relapse-remission" disease model, and realizing long-term drug use and precise disease management for patients. The goal is to achieve a new level of chronic disease management by scientifically improving long-term medication, precise disease management, and individualized medical treatment, effectively prolonging the remission period and reducing morbidity and disability rates.
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PURPOSE: To compare the difference and agreement in central corneal thickness (CCT), keratometry (K), anterior chamber depth (ACD), aqueous depth (AQD), and lens thickness (LT) measured with CASIA 2 and IOLMaster 700 in patients with cataract. METHODS: A total of 81 patients with cataract (81 eyes) scheduled for phacoemulsification were prospectively collected from March to May, 2020 in the cataract department of Zhongshan Ophthalmic Center, Sun Yat-sen University, including 43 males and 38 females with age of 61.5 ± 10.6 years. CCT, anterior Kf, anterior Ks, real Kf, real Ks, ACD, AQD, and LT were measured with CASIA 2 and IOLMaster 700. Paired t-test, intraclass correlation coefficients (ICCs), 95% limit of agreement (95% LoA), and Bland-Altman plots were performed and used to analyze the difference and agreement between the two devices. RESULTS: There was no statistically significant difference in anterior K measurement with the CASIA 2 (44.3 ± 1.66 mm) and IOLMaster 700 (44.31 ± 1.67 mm, P = 0.483). Differences among the CCT, anterior Kf, real Kf, real Ks, ACD, AQD, and LT measured by the two instruments were statistically significant (P < 0.001). The ICCs of CCT, anterior Kf, anterior Ks, real Kf, real Ks, ACD, AQD, and LT measurements between the two devices were 0.892, 0.991, 0.991, 0.827, 0.817, 0.937, 0.926, and 0.997, respectively. The 95% LoA between CASIA 2 and IOLMaster 700 was -30.06 to 0.43 µm for CCT, -0.3 to 0.48 D for anterior Kf, -0.46 to -0.43 D for anterior Ks, -1.49 to -0.49 D for real Kf, -1.62 to -0.49 D for Real Ks, -0.03 to 0.24 mm for ACD, 0.04 to 0.25 mm for AQD, and -0.06 to 0.09 mm for LT. CONCLUSION: Anterior Kf, anterior Ks, ACD, AQD, and LT have excellent agreement between the two devices. CCT, real Kf, and real Ks have moderate agreement between the two devices. It is recommended to use anterior Kf, anterior Ks, ACD, AQD, and LT interchangeably between CASIA 2 and IOLMaster 700.
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BACKGROUND AND AIM: To assess the reproducibility of the novel ultrasound biomicroscopy, Insight 100 and its agreement with a swept-source optical coherence tomography, CASIA2. METHODS: A total of 96 volunteers (96 eyes) were enrolled. The radius of anterior lens curvature (RAL), the radius of posterior lens curvature (RPL), lens thickness (LT), and lens diameter (LD) were measured with Insight 100 and CASIA2. A semiautomated software was used to adjust the measurement of LT (LTS) and LD (LDS) by Insight 100. Intraobserver and interobserver reproducibility of Insight 100 measurements, and the agreement of results from Insight 100 and CASIA2 were assessed with 95% limit of agreement (LoA), intraclass correlation coefficient (ICC), Pearson correlation, and linear regression. RESULTS: For Insight 100 measurements, the intraobserver ICCs of RAL, RPL, LTS, and LDS measurement were 0.996, 0.973, 0.936, and 0.889, and the interobserver ICCs were 0.987, 0.890, 0.974, and 0.816, respectively. There was an excellent correlation in LT measurements (R = 0.961, P < 0.001) but poor agreements in other parameters between the two devices. The LD measurements tended to be larger (95% CI: 0.768-0.928) in CASIA2 when compared with Insight 100. CONCLUSION: Insight 100 could obtain highly repeatable lens biometry in vivo. With better signal penetration, it shows promising potential in future clinical applications.
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PURPOSE: This study aimed to investigate the regulation of heme oxygenase-1 (HO-1) by paired box gene 6 (Pax6) and their roles in hydrogen peroxide (H2O2)-induced oxidative stress and apoptosis in lens epithelial cells (LECs) (SRA01/04, HLE-B3). METHODS: Lens anterior capsule membranes of mice of different ages were obtained to compare differences in the expression of Pax6 and HO-1 using Western blotting. Pax6-overexpressing plasmid and small interfering RNA were designed to overexpress and silence Pax6, respectively. Cobalt protoporphyrin (CoPP) was used to promote the expression of HO-1. Oxidative damage in LECs was induced by treatment with H2O2 (400 µM) for 24 h. Cell viability was measured using the Cell Counting Kit-8 assay. Intracellular reactive oxygen species (ROS) were detected using flow cytometry and immunofluorescence. Superoxide dismutase (SOD) level was measured using SOD Assay Kit and apoptotic cells were quantified using annexin V-fluorescein isothiocyanate/propidium iodide staining. RESULTS: Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs of mouse. Overexpressing Pax6 upregulated HO-1 expression level. Silencing Pax6 downregulated the HO-1 expression level, resulting in increased generation of ROS, reduced SOD activity, decreased cell viability, and increased apoptotic cells of LECs under H2O2-induced oxidative stress. Overexpressing Pax6 and CoPP both mitigates H2O2-induced oxidative stress by increasing the expression of HO-1 of LECs. CONCLUSION: Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs in mouse anterior capsules. Pax6 could regulate the expression of HO-1 in LECs. The decrease of Pax6 weakened the antioxidant ability of LECs under H2O2-induced oxidative stress by downregulating HO-1, which may be a potential mechanism for the formation of age-related cataract.
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Peróxido de Hidrogênio , Cristalino , Animais , Anexina A5/metabolismo , Antioxidantes/metabolismo , Apoptose , Cápsulas/metabolismo , Células Epiteliais/metabolismo , Fluoresceínas/metabolismo , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Peróxido de Hidrogênio/metabolismo , Isotiocianatos , Cristalino/metabolismo , Proteínas de Membrana , Camundongos , Estresse Oxidativo , Fator de Transcrição PAX6 , Propídio/metabolismo , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismoRESUMO
AIM: This study aimed to investigate the role of lncRNA MCM3AP-AS1 in diabetic retinopathy (DR). METHODS: Plasma MCM3AP-AS1 levels in DR patients (n = 80), T2DM patients (n = 80), and Controls (n = 80) were measured by qPCR and compared using ANOVA (one-way) and Tukey test. The expressions of lncRNA MCM3AP-AS1 and miR-211 in Human retinal pigment epithelial cells (hRPE) line ARPE-19 were detected by RT-qPCR. Western blot and annexin V-FITC staining were performed to investigate the role of MCM3AP-AS1/SIRT1 in ARPE-19 cell proliferation and apoptosis in vitro. RESULTS: We observed that MCM3AP-AS1 was downregulated in DR patients 25 comparing to T2D patients without significantly complications. Bioinformatics analysis showed that MCM3AP-AS1 might bind miR-211. However, no significant correlation between these two factors was observed in DR patients. Consistently, overexpression of MCM3AP-AS1 and miR-211 failed to affect the expression of each other in hRPE. Interestingly, MCM3AP-AS1 overexpression upregulated SIRT1, a target of miR-211. Moreover, MCM3AP-AS1 was downregulated in DR patients compared to type 2 diabetic mellitus patients without significant complications. In RPEs, high glucose treatment downregulated MCM3AP-AS1. Cell apoptosis analysis showed that MCM3AP-AS1 and SIRT1 overexpression decreased the apoptotic rate of RPEs, and miR-211 overexpression reduced the effect of MCM3AP-AS1 and SIRT1 overexpression. CONCLUSION: MCM3AP-AS1 is downregulated in DR and promotes cell apoptosis by regulating miR-211/SIRT1.
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PURPOSE: To investigate the prediction accuracy of intraocular lens (IOL) calculation formulas, and the impact of anterior chamber depth (ACD) and lens thickness (LT) measurement errors on IOL power calculation in patients undergoing combined phakic IOL (PIOL) removal and cataract surgery. DESIGN: Retrospective, consecutive case series study. METHODS: Thirty-six PIOL implanted eyes (12 anterior chamber PIOLs and 24 posterior chamber PIOLs [PC-PIOL]) undergoing cataract surgery were included. The prediction accuracy of new formulas (Barrett universal II, Emmetropia verifying optical, Kane, and Ladas super formula) and traditional formulas (Haigis, Hoffer Q, Holladay 1 and SRK/T) with or without Wang-Koch (WK) axial length (AL) adjustment was evaluated. The influence of ACD and LT measurement errors of IOLMaster 700 on refractive outcomes was also investigated. RESULTS: The Kane and traditional formulas with WK AL adjustment had no significant systematic prediction error and displayed a smaller median absolute error, whereas the other formulas showed significant hyperopia shift (P < .05) and relatively lower prediction accuracy. The accuracy rate of IOLMaster 700 in measuring the ACD and LT was 100% in eyes with anterior chamber PIOL implantation, and 37.50% in the PC-PIOL subgroup. No significant difference was observed in refractive outcomes of formulas using correct and wrong parameters in the PC-PIOL subgroup (P > .05). CONCLUSIONS: The Kane and traditional formulas with WK AL adjustment exhibited relatively higher prediction accuracy in patients who underwent combined PIOL removal and cataract surgery. The IOLMaster 700 displayed low accuracy in ACD and LT measurements for PC-PIOL implanted eyes, but showed negligible impact on IOL prediction accuracy.
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Catarata , Lentes Intraoculares , Facoemulsificação , Lentes Intraoculares Fácicas , Comprimento Axial do Olho , Biometria , Humanos , Implante de Lente Intraocular , Óptica e Fotônica , Refração Ocular , Estudos RetrospectivosRESUMO
OBJECTIVE: To investigate the protective efficacies and potent mechanisms of combination therapy with semaglutide and rosiglitazone (RSG) on the high-glucose incubated human ARPE-19 cells and diabetic retinopathy (DR) model rats. MAIN METHODS: The CCK-8 methods were used to evaluate the protective effects of semaglutide and RSG alone or combination on the cell viability of high-glucose treated ARPE-19 cells. After the DR rat model was established, the effects of combined treatment on general indexes, retinal morphological changes, retinal Müller cells as well as PI3K/Akt/MTOR related factors of DR model rats were investigated. RESULTS: The CCK-8 assay showed obviously enhanced protective efficacies of combination therapy with semaglutide and RSG on the ARPE-19 with oxidative stress induced by high-glucose with combination index all below 1.5 demonstrating obvious synergistic effects. Combined incubation could also effectively decrease the expression of inflammatory factors, including TNF-α, IL-1ß, IL-6, and the increase of ROS content in ARPE cell culture supernatant induced by high-glucose. Combined use of the antioxidant, PI3K/Akt and mTOR inhibitors, we further demonstrated that combined incubation of semaglutide and RSG could effectively by reduce high glucose-induced inflammatory injury inhibiting ROS/PI3K/Akt/mTOR signaling. Furthermore, chronic combination treatment effectively improved the histopathological characteristics and down-regulated the GFAP expression in Müller cells as well as PI3K/Akt/MTOR signaling pathway-related factors in retina which was better than any monomer treatment group. CONCLUSIONS: Combined semaglutide with RSG exhibited synergistically protective efficacies on retinal cells by decreasing the GFAP expression, inhibiting oxidative stress and PI3K/Akt/MTOR signaling-transduction in DR model rats.
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Retinopatia Diabética/tratamento farmacológico , Peptídeos Semelhantes ao Glucagon/uso terapêutico , Rosiglitazona/uso terapêutico , Animais , Antioxidantes/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Quimioterapia Combinada , Células Ependimogliais/efeitos dos fármacos , Células Ependimogliais/metabolismo , Células Ependimogliais/patologia , Proteína Glial Fibrilar Ácida/metabolismo , Peptídeos Semelhantes ao Glucagon/farmacologia , Humanos , Inflamação/patologia , Masculino , Fosfatidilinositol 3-Quinases/metabolismo , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Sprague-Dawley , Rosiglitazona/farmacologia , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismoRESUMO
INTRODUCTION: Demodex and bacteria are both components of the ocular surface micro-ecology, constituting a complex interaction. This study aims to explore how ocular surface Demodex infestation (DI) affects ocular surface microbial communities and diversity. METHODS: We recruited 255 subjects, and examined the correlation between ocular surface mite infestation and clinical indicators such as age, blood glucose level, dry eye symptoms, and blood pressure. 16S rRNA sequencing was performed on the conjunctival swab samples of 14 patients with ocular DI (P group) and 17 healthy people (N group). For further analysis, the subjects were divided into four subgroups, i.e. N-NMGD (n = 11), N-MGD (n = 6), P-NMGD (n = 6), and P-MGD (n = 8), according to meibomian gland dysfunction (MGD) or no MGD (NMGD). RESULTS: There was no difference in the α-diversity of ocular surface microbial communities between the DI and healthy control groups. In linear discriminant analysis effect size (LEfSe), there were more Acinetobacter, Novosphingobium, and Anoxybacillus in the DI group and fewer Novosphingobium, Lactobacillus, and Candidatus Microthrix in the healthy control group. P-NMGD had more Thermaceae and fewer Pseudomonas than P-MGD. There were more Bacteroidetes in N-NMGD than in N-MGD. The α-diversity of P-NMGD was lower than that of N-NMGD (Shannon index, P = 0.027). At the same time, the α-diversity of N-MGD was lower than that of N-NMGD (Shannon, Simpson, and dominance index, P = 0.048). There was no significant difference in ß-diversity or in the primary flora at the phylum and genus levels between groups and subgroups. CONCLUSION: DI had no significant effect on the diversity of ocular surface microbial communities. DI primarily changed the dominant flora and relative abundance of ocular surface microbial communities. MGD may play an important role in this process.
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The relation between microRNAs (miRNAs) and malignant melanoma has been demonstrated in previous studies, while there was little research about miR-139-5p and malignant melanoma. The aim of this study is to investigate the ability of miR-139-5p in malignant melanoma cells via the modulation of the PI3K/AKT signaling pathway by targeting IGF1R. MiR-139-5p expression in malignant melanoma tissues and 5 malignant melanoma cell lines was detected. The melanoma cells were transfected with miR-139-5p mimic negative control (NC) sequence, miR-139-5p mimic, IGF1R overexpressed recombinant plasmid NC or IGF1R overexpressed sequence. The expression of Akt signaling pathway-related protein was evaluated. The biological functions in malignant melanoma cells were evaluated by a string of experiments. MiR-139-5p expressed a poor level in tissues and cell lines of malignant melanoma. Overexpressed miR-139-5p suppressed the cell proliferation, migration, and invasion, and contributed to the promoted apoptosis of malignant melanoma cells by decreasing IGF1R. MiR-139-5p down-regulated the IGF1R expression, and IGF1R accelerated the activation of the PI3K/AKT signaling pathway. miR-139-5p reversed the promotive impacts of IGF1R on the PI3K/AKT signaling pathway. The study validates that miR-139-5p could suppress malignant melanoma progression through the repression of the PI3K/AKT signaling pathway by down-regulating IGF1R. Therefore, miR-139-5p could pave a new way for the treatment of malignant melanoma.
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Proliferação de Células/genética , Melanoma/genética , MicroRNAs/genética , Receptor IGF Tipo 1/genética , Adulto , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Feminino , Humanos , Masculino , Melanoma/patologia , Pessoa de Meia-Idade , Metástase Neoplásica , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/genéticaRESUMO
Our study aimed to investigate the role of long non-coding RNAs (lncRNA) TP73-AS1 in retinoblastoma (Rb). In the present study, we found that TP73-AS1 was up-regulated, while miR-139-3p was down-regulated in Rb. TP73-AS1 and miR-139-3p were inversely correlated in Rb tissues. In cells of Rb cell lines, overexpression of miR-139-3p failed to affect TP73-AS1, while TP73-AS1 overexpression caused the down-regulated miR-139-3p TP73-AS1 overexpression caused promoted proliferation of Rb cells but showed no significant effects on cell migration and invasion. miR-139-3p overexpression played an opposite role and attenuated the effects of TP73-AS1 overexpression. Therefore, lncRNA TP73-AS1 may down-regulate miR-139-3p to promote Rb cell proliferation.
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Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , RNA Longo não Codificante/genética , Neoplasias da Retina/genética , Retinoblastoma/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células/genética , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , MicroRNAs/metabolismo , Invasividade Neoplásica , RNA Longo não Codificante/metabolismo , Neoplasias da Retina/metabolismo , Neoplasias da Retina/patologia , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Transdução de SinaisAssuntos
Neurocitoma/patologia , Neoplasias da Medula Espinal/patologia , Antígenos Nucleares/metabolismo , Cordotomia/métodos , Diagnóstico Diferencial , Ependimoma/metabolismo , Ependimoma/patologia , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas do Tecido Nervoso/metabolismo , Neurocitoma/metabolismo , Neurocitoma/cirurgia , Oligodendroglioma , Proteínas S100/metabolismo , Neoplasias da Medula Espinal/metabolismo , Neoplasias da Medula Espinal/cirurgia , Sinaptofisina/metabolismoRESUMO
The aim of the present study was to investigate the expression levels of Yes-associated protein (YAP) in different grades of laryngeal squamous cell carcinoma (LSCC) tissues and vocal cord polyps tissues, and to investigate any correlations with clinical factors. The expression of YAP in 128 cases of LSCC and 10 cases of vocal cord polyps tissues was tested using immunohistochemistry. YAP was primarily present in the nucleus of LSCC and controls, whereas phosphorylated YAP expression was present in the cytoplasm. The results indicated that YAP expression was upregulated in LSCC samples compared with vocal cord polyps tissues. YAP expression was positively correlated with the malignant degree of LSCC (P<0.01) and a high level of YAP expression in LSCC tissues was correlated with pathological type, lymphatic metastasis and clinical stage. The present study provided evidence for the expression and localization of YAP in LSCC and vocal cord polyps tissues. Thus, YAP may be involved in the occurrence and development of LSCC as an oncogene.
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Primary intestinal extranodal NK/T-cell lymphoma, nasal type, is an extremely rare type of lymphoma with poor prognosis, and early diagnosis is challenging. Here we have investigated the clinicopathologic features and immunophenotypes of primary intestinal extranodal NK/T cell lymphomas, nasal type, in 10 Chinese patients. Complete staging data showed that 1 patient had stage I disease, 7 had stage II disease, and 2 had stage III disease. Eight of 10 (80%) patients had lymphadenopathy and none had bone marrow involvement. All the patients had a low International Prognostic Index (IPI) score (<3) at presentation. The median age at the time of diagnosis was 37.5 years (range = 24-68 years). All the patients died within 21 months, and the median survival time was 9.5 months (range = 2-21 months). So the conventional IPI and staging system failed to predict the outcome of the patients with the lymphoma. Except for the size of tumor cells, most of the morphologic features of the cases we studied were similar to those involving the midline facial tissue. Immunohistochemical studies showed the expression of cytotoxic markers (100%), CD2 (100%), CD3ε (90%), CD56 (80%), P53 (60%), CD30 (30%), LMP1 (30%), EBNA3A (0%). Nine cases (90%) highly expressed Ki-67. In situ hybridization for Epstein-Barr virus-encoded small RNA was positive in all cases.
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Biomarcadores Tumorais/análise , Linfoma Extranodal de Células T-NK/patologia , Adulto , Idoso , Feminino , Humanos , Imuno-Histoquímica , Imunofenotipagem , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
OBJECTIVE: To detect the gene expression of Pax-6 in cultured lens epithelial cells (LECs) of mouse in vitro in order to study the gene factor that maintains the LECs characteristics. METHODS: Cultured the LECs (passage 1 to 3) were used in the study, the expression of Pax-6 gene in LECs was demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR), immunohistochemical staining and Western blot. RESULTS: The mRNA of Pax-6 gene was positive in primary culture, the first and second passage of LECs using RT-PCR. The protein of Pax-6 gene was also positive in Western blot and immunohistochemical staining, but it was weak positive in the third passage of LECs and none in all passages of negative control groups. CONCLUSION: The LECs are positive for the gene of Pax-6. The normal expression of Pax-6 gene is vital for maintaining the LECs characteristics.