ABSTRACT
OBJECTIVE: Nordalbergin is a coumarin extracted from Dalbergia sissoo DC. To date, the biological effects of nordalbergin have not been well investigated. To investigate the anti-inflammatory responses and the anti-oxidant abilities of nordalbergin using lipopolysaccharide (LPS)-activated macrophages and LPS-induced sepsis mouse model. MATERIALS AND METHODS: Production of nitrite oxide (NO), prostaglandin E2 (PGE2), pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1ß), reactive oxygen species (ROS), tissue damage and serum inflammatory markers, and the activation of the NLRP3 inflammasome were examined. RESULTS: Our results indicated that nordalbergin reduced the production of NO and pro-inflammatory cytokines in vitro and ex vivo. Nordalbergin also suppressed iNOS and cyclooxygenase-2 expressions, decreased NF-κB activity, and attenuated MAPKs signaling pathway activation by decreasing JNK and p38 phosphorylation by LPS-activated J774A.1 macrophages. Notably, nordalbergin diminished NLRP3 inflammasome activation via repressing the maturation of IL-1ß and caspase-1 and suppressing ROS production by LPS/ATP- and LPS/nigericin-activated J774A.1 macrophages. Furthermore, nordalbergin exhibited protective effects against the infiltration of inflammatory cells and also inhibited the levels of organ damage markers (AST, ALT, BUN) by LPS-challenged mice. CONCLUSION: Nordalbergin possesses anti-inflammatory effects in macrophage-mediated innate immune responses, alleviates ROS production, decreases NLRP3 activation, and exhibits protective effects against LPS-induced tissue damage in mice.
ABSTRACT
Previous studies have highlighted the importance of outdoor time in reducing the risk of myopia progression. Although ultraviolet A (UVA) radiation dominates in terms of energy with respect to the UV radiation reaching the Earth's surface, its effects on the exposed anterior sclera have not been well studied. This study was designed to investigate the UVA-induced biological effects at peak sunlight levels in human scleral fibroblasts (HSFs). Using next-generation sequencing (NGS), we analyzed the differentially expressed genes (DEGs) in UVA-treated and normal HSFs. Further, we then identified the functions and key regulators of the DEGs using bioinformatics analysis, and verified the effects of UVA on gene and protein expression in HSFs using real-time PCR, western blotting, and immunofluorescence imaging. The highest level of solar UVA (365 nm) was 3.4 ± 0.18 (mW/cm2). The results from the functional analysis of the DEGs were related to structural changes in the extracellular matrix (ECM) and protein metabolism. Transforming growth factor-ß1 (TGF-ß1) and Smad3 were predicted to be potential upstream regulators, associated with ECM organization. Exposure to a single wavelength of UVA (365 nm, 3 mW/cm2) for 1 h for 5 consecutive days induced the downregulation of the mRNA of ECM genes including COL1A1, COL3A1, COL5A1, VCAN and collagen I protein in HSF. UVA downregulated Smad3 protein and reduced TGF-ß-induced collagen I protein production following UVA exposure in HSF. In conclusion, high UVA exposure reduces TGF-ß signaling and collagen I production by modulating Smad levels in HSF. The effects of overexposure to high-intensity UVA on myopia control require further investigations.
Subject(s)
Myopia , Transforming Growth Factor beta , Humans , Transforming Growth Factor beta/metabolism , Sclera/metabolism , Collagen/metabolism , Transforming Growth Factor beta1/metabolism , Collagen Type I/genetics , Collagen Type I/metabolism , Fibroblasts/metabolism , Ultraviolet Rays/adverse effects , Myopia/metabolism , Transforming Growth Factors/metabolism , Transforming Growth Factors/pharmacologyABSTRACT
Microglia-associated neuroinflammation is recognized as a critical factor in the pathogenesis of neurodegenerative diseases; however, there is no effective treatment for the blockage of neurodegenerative disease progression. In this study, the effect of nordalbergin, a coumarin isolated from the wood bark of Dalbergia sissoo, on lipopolysaccharide (LPS)-induced inflammatory responses was investigated using murine microglial BV2 cells. Cell viability was assessed using the MTT assay, whereas nitric oxide (NO) production was analyzed using the Griess reagent. Secretion of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) was detected by the ELISA. The expression of inducible NO synthase (iNOS), cyclooxygenase (COX)-2, mitogen-activated protein kinases (MAPKs) and NLRP3 inflammasome-related proteins was assessed by Western blot. The production of mitochondrial reactive oxygen species (ROS) and intracellular ROS was detected using flow cytometry. Our experimental results indicated that nordalbergin ≤20 µM suppressed NO, IL-6, TNF-α and IL-1ß production; decreased iNOS and COX-2 expression; inhibited MAPKs activation; attenuated NLRP3 inflammasome activation; and reduced both intracellular and mitochondrial ROS production by LPS-stimulated BV2 cells in a dose-dependent manner. These results demonstrate that nordalbergin exhibits anti-inflammatory and anti-oxidative activities through inhibiting MAPK signaling pathway, NLRP3 inflammasome activation and ROS production, suggesting that nordalbergin might have the potential to inhibit neurodegenerative disease progression.
Subject(s)
Lipopolysaccharides , Neurodegenerative Diseases , Mice , Animals , Lipopolysaccharides/pharmacology , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Inflammasomes/metabolism , Microglia/metabolism , Reactive Oxygen Species/metabolism , Neuroinflammatory Diseases , Interleukin-6/metabolism , Tumor Necrosis Factor-alpha/metabolism , Neurodegenerative Diseases/metabolism , Signal Transduction , Mitogen-Activated Protein Kinases/metabolism , NF-kappa B/metabolismABSTRACT
This study has two novel findings: it is not only the first to deduct potential genes involved in scleral growth repression upon atropine instillation from a prevention point of view, but also the first to demonstrate that only slight changes in scleral gene expression were found after atropine treatment as side effects and safety reasons of the eye drops are of concern. The sclera determines the final ocular shape and size, constituting of scleral fibroblasts as the principal cell type and the major regulator of extracellular matrix. The aim of our study was to identify differentially expressed genes and microRNA regulations in atropine-treated scleral fibroblasts that are potentially involved in preventing the onset of excessive ocular growth using next-generation sequencing and bioinformatics approaches. Differentially expressed genes were functionally enriched in anti-remodeling effects, comprising of structural changes of extracellular matrix and metabolic pathways involving cell differentiation. Significant canonical pathways were correlated to inhibition of melatonin degradation, which was compatible with our clinical practice as atropine eye drops are instilled at night. Validation of the dysregulated genes with previous eye growth-related arrays and through microRNA-mRNA interaction predictions revealed the association of hsa-miR-2682-5p-KCNJ5 and hsa-miR-2682-5p-PRLR with scleral anti-remodeling and circadian rhythmicity. Our findings present new insights into understanding the anti-myopic effects of atropine, which may assist in prevention of myopia development.
Subject(s)
Atropine/pharmacology , Myopia/drug therapy , Sclera/drug effects , Transcriptome/genetics , Circadian Rhythm/drug effects , Computational Biology , Extracellular Matrix/genetics , Fibroblasts/drug effects , G Protein-Coupled Inwardly-Rectifying Potassium Channels/genetics , Gene Expression Regulation/drug effects , Gene Regulatory Networks/genetics , High-Throughput Nucleotide Sequencing , Humans , MicroRNAs/genetics , Myopia/genetics , Myopia/pathology , RNA, Messenger/genetics , Sclera/growth & development , Sclera/pathologyABSTRACT
BACKGROUND: The aim of this study was to measure the changes in the bacterial bioburden in orthokeratology (OK) lens storage cases using the DNA dot hybridization assay (DHA) after forewarning patients about their bacterial contamination severity. METHODS: Thirty-one OK lens wearers were prospectively enrolled in this study. Dot hybridization assay was used for serial measurements of bacterial bioburden in OK storage cases after lenses had been soaked for approximately 6 hr. After the first assessment, the lens wearers were informed of the extent of case contamination and the possible risk of microbial keratitis (MK), and best practices for lens care and lens case hygiene were reviewed and reinforced. A second assessment by the same DHA method was performed after approximately 6 months. RESULTS: Two universal bacterial probes confirmed a significant decrease in bacterial bioburden at the second assessment (P<0.01 and P<0.001). Genus-specific probes showed significant reductions in Acinetobacter and Klebsiella (P=0.02 and P=0.01), but not in Pseudomonas (P=0.42). CONCLUSIONS: Making OK lens wearers aware of the bacterial bioburden in their lens cases resulted in improved quality of case care and reduced bioburden. Our results suggest that a strategy of bioburden assessment with forewarning could be a useful method to decrease the incidence of OK-related MK.
Subject(s)
Bacteria/genetics , Contact Lenses/microbiology , DNA, Bacterial/analysis , DNA/analysis , Eye Infections, Bacterial/prevention & control , Nucleic Acid Hybridization/methods , Optical Storage Devices , Adolescent , Bacteria/isolation & purification , Child , Contact Lens Solutions/pharmacology , DNA Probes , Equipment Contamination , Eye Infections, Bacterial/microbiology , Female , Humans , Male , Orthokeratologic Procedures , Prospective StudiesABSTRACT
PURPOSE: To develop a PCR gel analysis method for assessing the bacterial bioburden in orthokeratology contact lens (OK) case fluid determined by culture. METHODS: A prospective study with the participation of 41 OK wearers (20 girls, 21 boys) was performed. The mean OK-wearing experience was 3.5±1.9 years. PCR was used to assess the bacterial bioburden (colony-forming units per milliliter) of OK after removal and soaking in the storage case for 6 h. The signal intensity of the PCR bands was analyzed after grayscale image transformation. The difference (cPCR-d) and ratio (cPCR-r) between a PCR signal and its background were used as two standardized indices of PCR signals. The association between the two indices of the PCR signals and the bacterial bioburden determined by culture were analyzed with Pearson's correlation coefficient (r) and receiver operating characteristic (ROC) plots. RESULTS: At least one microbe was isolated from the OK lens case from 38 of the 41 subjects. Both cPCR-d and cPCR-r showed strong correlations with the bacterial bioburden (r>0.7, p<0.0001). ROC analysis enabled good determination of the cutoff values for the two PCR indices with acceptable sensitivity and specificity (78-89%) to assess the degree of bacterial contamination. CONCLUSIONS: The high microbial contamination rate of the OK lens cases revealed the general inappropriate lens care by OK wearers. PCR analysis provides an alternative and rapid method for assessing the bacterial bioburden of OK lens cases, and these results should serve as a warning to OK wearers to follow appropriate lens care procedures to prevent infection.
Subject(s)
Bacteria/isolation & purification , Contact Lenses/microbiology , DNA, Bacterial/analysis , Equipment Contamination , Orthokeratologic Procedures/instrumentation , Product Packaging , Adolescent , Bacterial Physiological Phenomena , Biofilms/growth & development , Child , Female , Humans , Male , Polymerase Chain Reaction , Prospective StudiesABSTRACT
OBJECTIVE: To assess the bioburden in an orthokeratology contact lens (OK) care system (defined by microbial identification from OK case fluid) and to identify the risk factors causing high bioburden for pediatric OK wearers in southern Taiwan. METHODS: A prospective study for the investigation of bioburden in the OK care system was performed in a tertiary medical center in southern Taiwan. Microbial isolates from the case fluids soaking OKs were analyzed, and pathogenicity was determined. Age, gender, OK experiences, and contact lens care habits were considered the potential risk factors of microbial bioburden (colony-forming units per milliliter) for causal analysis. RESULTS: Forty-one OK wearers (20 female and 21 male subjects) participated in this study. The mean age was 12.7 years, and the mean OK-wearing experience was 3.5 years. A total of 86 microbial strains were isolated from 38 culture-positive specimens. Frequently reported pathogens in contact lens-related microbial keratitis were less common in the current study, but still present, including 4 strains (5%) of Serratia marcescens, 1 strain (1%) of Pseudomonas aeruginosa, and 1 strain (1%) of Staphylococcus aureus. Microbial bioburden of the OK care system was significantly higher (P<0.05) in male subjects. CONCLUSIONS: The contamination rate of the OK care system was high, and many isolated microorganisms had potential pathogenicity. Reinforcement of proper contact lens care and education should be mandatory for OK wearers, particularly for male subjects, to decrease the risk of high bioburden of the OK care system.
Subject(s)
Bacteria/isolation & purification , Contact Lens Solutions , Contact Lenses, Hydrophilic/microbiology , Equipment Contamination/statistics & numerical data , Orthokeratologic Procedures , Adolescent , Child , Eye Infections/etiology , Female , Humans , Male , Orthokeratologic Procedures/adverse effects , Orthokeratologic Procedures/methods , Prospective Studies , Risk Factors , TaiwanABSTRACT
Aminoglycosides exhibit relatively poor activity against intracellular Salmonella enterica serovar Typhimurium due to their low permeativity across eukaryotic cell membranes. Previously, we identified the unique ability of AR-12, a celecoxib-derived small-molecule agent, to eradicate intracellular Salmonella Typhimurium in macrophages by facilitating autophagosome formation and suppressing Akt kinase signaling. In light of this unique mode of antibacterial action, we investigated the ability of AR-12 to sensitize intracellular Salmonella to aminoglycosides in macrophages and in an animal model. The antibacterial activities of AR-12 combined with various aminoglycosides, including streptomycin, kanamycin, gentamicin, and amikacin, against intracellular S. Typhimurium in murine RAW264.7 macrophages were assessed. Cells were infected with S. Typhimurium followed by treatment with AR-12 or individual aminoglycosides or with combinations for 24 h. The in vivo efficacies of AR-12, alone or in combination with gentamicin or amikacin, were also assessed by treating S. Typhimurium-infected BALB/c mice daily for 14 consecutive days. Exposure of S. Typhimurium-infected RAW264.7 cells to a combination of AR-12 with individual aminoglycosides led to a reduction in bacterial survival (P < 0.05), both intracellular and extracellular, that was greater than that seen with the aminoglycosides alone. This sensitizing effect, however, was not associated with increased aminoglycoside penetration into bacteria or macrophages. Moreover, daily intraperitoneal injection of AR-12 at 0.1 mg/kg of body weight significantly increased the in vivo efficacy of gentamicin and amikacin in prolonging the survival of S. Typhimurium-infected mice. These findings indicate that the unique ability of AR-12 to enhance the in vivo efficacy of aminoglycosides might have translational potential for efforts to develop novel strategies for the treatment of salmonellosis.
Subject(s)
Aminoglycosides/pharmacology , Anti-Bacterial Agents/pharmacology , Pyrazoles/pharmacology , Salmonella Infections, Animal/drug therapy , Salmonella typhimurium/drug effects , Sulfonamides/pharmacology , Amikacin/pharmacology , Animals , Drug Administration Schedule , Drug Synergism , Drug Therapy, Combination , Female , Gentamicins/pharmacology , Injections, Intraperitoneal , Kanamycin/pharmacology , Macrophages/drug effects , Macrophages/microbiology , Mice , Mice, Inbred BALB C , Microbial Sensitivity Tests , Microbial Viability , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/growth & development , Streptomycin/pharmacology , Survival AnalysisABSTRACT
BACKGROUND: During the COVID-19 pandemic period, it was difficult to carry out regular and scheduled follow-up of patients in the outpatient department, especially during lockdown periods. However, early detection of initial infection or other serious conditions is vital for patients after ocular surgery, such as intravitreal injection (IVI) for age-related macular degeneration (AMD). OBJECTIVE: We evaluated the use of a smartphone-based postoperative care chatbot system (PCCS) with an instant bidirectional feedback system for patients to self-report postoperative symptoms and signs. METHODS: During the COVID-19 level 3 epidemic alert in July 2021 in Taiwan, the PCCS alerted the patients to report and grade 6 ocular symptoms and signs associated with ocular inflammation or retinal detachment. Patients used the PCCS for 7 days post surgery to assess their symptoms and signs each day after receiving an alert. Data were automatically collected using a cloud computer system, including symptom grades and messages sent to medical staff for further medical assistance. A user satisfaction questionnaire was administered to the patients on the seventh day post surgery. RESULTS: In total, 185 patients participated in this study. There were 26 (3.03%) reports of symptom grade deterioration (including increased blurred vision, eye swelling, nausea, and floaters or flashes) from 12 (6.5%) patients. We found no difference in the gender of patients who received an early medical consultation. One case of endophthalmitis was reported, wherein an improvement was observed after prompt administration of IVI antibiotics twice. Overall, 87% (n=185) of patients were satisfied or very satisfied with communicating their symptoms instantly through the app; they were willing to use it again and believed that it could improve the quality of care. Seven of the 185 (3.8%) patients had an earlier medical consultation and 1 (0.5%) had endophthalmitis. CONCLUSIONS: The chatbot system, designed for self-reporting postoperative symptoms and providing instant bidirectional feedback on smartphones, could be beneficial for enhancing the quality of care in early medical consultations without gender differences among patients with AMD receiving IVI, and achieved satisfactory responses from patients.
ABSTRACT
Glaucine is an aporphine alkaloid with anti-inflammatory, bronchodilator and anti-cancer activities. However, the effects of glaucine in the regulation of age-related macular degeneration (AMD) remain unclear. Herein, we aimed to investigate the anti-angiogenetic and anti-inflammatory effects of glaucine in ARPE-19 cells. ARPE-19 cells were treated with N-(methoxyoxoacetyl)-glycine, methyl ester (DMOG) and cobalt chloride (CoCl2) for induction of hypoxia, while lipopolysaccharide (LPS) treatment was used for elicitation of inflammatory response. Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The expression of hypoxia-inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF) were measured by Western blot. The secretion of VEGF, interleukin (IL)-6 and monocyte chemoattractant protein-1 (MCP-1) was detected using enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were used for tube formation analysis. Expression of HIF-1α and secretion of VEGF were significantly increased under DMOG and CoCl2 induction, whereas glaucine significantly attenuated both HIF-1α expression and VEGF secretion by DMOG- and CoCl2-induced ARPE-19 cells. In addition, glaucine suppressed the tube formation by DMOG- and CoCl2-induced HUVEC cells. Moreover, glaucine also attenuated the production of IL-6 and MCP-1 by LPS-induced ARPE-19 cells. This study indicated that glaucine exhibited anti-angiogenic and anti-inflammatory effects, suggesting that glaucine might have benefits for the treatment of AMD.
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Optic pathway glioma (OPG) is one of the causes of pediatric visual impairment. Unfortunately, there is as yet no cure for such a disease. Understanding the underlying mechanisms and the potential therapeutic strategies may help to delay the progression of OPG and rescue the visual morbidities. Here, we provide an overview of preclinical OPG studies and the regulatory pathways controlling OPG pathophysiology. We next discuss the role of microenvironmental cells (neurons, T cells, and tumor-associated microglia and macrophages) in OPG development. Last, we provide insight into potential therapeutic strategies for treating OPG and promoting axon regeneration.
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Glaucoma is a neurodegenerative eye disease that causes permanent vision impairment. The main pathological characteristics of glaucoma are retinal ganglion cell (RGC) loss and optic nerve degeneration. Glaucoma can be caused by elevated intraocular pressure (IOP), although some cases are congenital or occur in patients with normal IOP. Current glaucoma treatments rely on medicine and surgery to lower IOP, which only delays disease progression. First-line glaucoma medicines are supported by pharmacotherapy advancements such as Rho kinase inhibitors and innovative drug delivery systems. Glaucoma surgery has shifted to safer minimally invasive (or microinvasive) glaucoma surgery, but further trials are needed to validate long-term efficacy. Further, growing evidence shows that adeno-associated virus gene transduction and stem cell-based RGC replacement therapy hold potential to treat optic nerve fiber degeneration and glaucoma. However, better understanding of the regulatory mechanisms of RGC development is needed to provide insight into RGC differentiation from stem cells and help choose target genes for viral therapy. In this review, we overview current progress in RGC development research, optic nerve fiber regeneration, and human stem cell-derived RGC differentiation and transplantation. We also provide an outlook on perspectives and challenges in the field.
Subject(s)
Glaucoma , Neurodegenerative Diseases , Optic Nerve Diseases , Humans , Animals , Glaucoma/drug therapy , Glaucoma/pathology , Retinal Ganglion Cells/pathology , Optic Nerve Diseases/therapy , Optic Nerve Diseases/pathology , Disease Progression , Neurodegenerative Diseases/pathology , Disease Models, AnimalABSTRACT
Kurarinone, a major lavandulyl flavanone found in the roots of Sophora flavescens aiton, has been reported to exhibit anti-inflammatory and anti-oxidative activities in lipopolysaccharide (LPS)-induced macrophages; however, the effects of kurarinone on the activation of NLRP3 inflammasome and the protective effects against sepsis have not been well investigated. In this study, we aimed to investigate the impacts of kurarinone on NLRP3 inflammasome activation in lipopolysaccharide (LPS)-induced macrophages and its protective effects against sepsis in vivo. Secretion of pro-inflammatory cytokines, activation of MAPKs and NF-κB signaling pathways, formation of NLRP3 inflammasome, and production of reactive oxygen species (ROS) by LPS-induced macrophages were examined; additionally, in vivo LPS-induced endotoxemia model was used to investigate the protective effects of kurarinone in sepsis-induced damages. Our experimental results demonstrated that kurarinone inhibited the expression of iNOS and COX-2, suppressed the phosphorylation of MAPKs, attenuated the production of TNF-α, IL-6, nitric oxide (NO) and ROS, repressed the activation of the NLRP3 inflammasome, and impeded the maturation and secretion of IL-1ß and caspase-1. Furthermore, the administration of kurarinone attenuated the infiltration of neutrophils in the lung, kidneys and liver, reduced the expression of organ damage markers, and increased the survival rate in LPS-challenged mice. Collectively, our study demonstrated that kurarinone can protect against LPS-induced sepsis damage and exert anti-inflammatory effects via inhibiting MAPK/NF-κB pathways, attenuating NLRP3 inflammasome formation, and preventing intracellular ROS accumulation, suggesting that kurarinone might have potential for treating sepsis and inflammation-related diseases.
Subject(s)
Inflammasomes , Sepsis , Mice , Animals , Inflammasomes/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Lipopolysaccharides/toxicity , Reactive Oxygen Species/metabolism , NF-kappa B/metabolism , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Sepsis/chemically induced , Sepsis/drug therapyABSTRACT
BACKGROUND: Retinoblastoma, the most common pediatric intraocular malignancy, can develop during embryogenesis, with most children being diagnosed at 3-4 years of age. Multimodal therapies are typically associated with high levels of cytotoxicity and side effects. Therefore, the development of novel treatments with minimal side effects is crucial. Magnolol has a significant anti-tumor effect on various cancers. However, its antitumor effect on retinoblastoma remains unclear. PURPOSE: The study aimed to determine the effects of magnolol on the regulation of EMT, migration, invasion, and cancer progression in retinoblastoma and the modulation of miR-200c-3p expression and the Wnt/ zinc finger E-box binding homeobox 1 (ZEB1)/E-cadherin axis in vivo and in vitro. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay was used to evaluate magnolol-induced cell toxicity in the Y79 retinoblastoma cell line. Flow cytometry and immunostaining assays were performed to investigate the magnolol-regulated mitochondrial membrane potential and the intracellular and mitochondrial reactive oxygen species levels in Y79 retinoblastoma cells. Orthotopic and subcutaneous xenograft experiments were performed in eight-week-old male null mice to study retinoblastoma progression and metastasis. In situ hybridization and quantitative reverse transcription polymerase chain reaction (RT-qPCR) assays were performed to evaluate the level of the anti-cancer miRNA miR-200c-3p. The mRNA and protein levels of E-cadherin, ß-catenin, α-smooth muscle actin (α-SMA), fibronectin-1, and ZEB1 were analyzed using RT-qPCR, immunoblot, immunocytochemistry, and immunohistochemistry assays in vitro and in vivo. RESULTS: Magnolol increased E-cadherin levels and reduced the activation of the EMT signaling pathway, EMT, tumor growth, metastasis, and cancer progression in the Y79 retinoblastoma cell line as well as in the orthotopic and subcutaneous xenograft animal models. Furthermore, magnolol increased the expression of miR-200c-3p. Our results demonstrate that miRNA-200c-3p inhibits EMT progression through the Wnt16/ß-catenin/ZEB1/E-cadherin axis, and the ZEB1 silencing response shows that miR-200c-3p regulates ZEB1-mediated EMT in retinoblastoma. CONCLUSION: Magnolol has an antitumor effect by increasing E-cadherin and miRNA-200c-3p expression to regulate ZEB1-mediated EMT and cancer progression in retinoblastoma. The anti-tumor effect of magnolol by increasing E-cadherin and miRNA-200c-3p expression to regulate ZEB1-mediated EMT and cancer progression in retinoblastoma has been elucidated for the first time.
Subject(s)
MicroRNAs , Retinal Neoplasms , Retinoblastoma , Animals , Mice , Humans , Male , Epithelial-Mesenchymal Transition/genetics , Retinoblastoma/drug therapy , Retinoblastoma/genetics , Cell Line, Tumor , MicroRNAs/genetics , MicroRNAs/metabolism , Cadherins/metabolism , Retinal Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Cell Movement/genetics , Zinc Finger E-box-Binding Homeobox 1/genetics , Zinc Finger E-box-Binding Homeobox 1/metabolismABSTRACT
Purpose: Neurodegenerative diseases are associated with neuroinflammation along with activation of microglia and oxidative stress, but currently lack effective treatments. Punicalagin is a natural bio-sourced product that exhibits anti-inflammatory effects on several chronic diseases; however, the anti-inflammatory and anti-oxidative effects on microglia have not been well examined. This study aimed to investigate the effects of punicalagin on LPS-induced inflammatory responses, NLRP3 inflammasome activation, and the production of ROS using murine microglia BV2 cells. Methods: BV2 cells were pre-treated with punicalagin following LPS treatment to induce inflammation. The secretion of NO and PGE2 was analyzed by Griess reagent and ELISA respectively, while the expressions of iNOS, COX-2, STAT3, ERK, JNK, and p38 were analyzed using Western blotting, the production of IL-6 was measured by ELISA, and the activity of NF-κB was detected using promoter reporter assay. To examine whether punicalagin affects NLRP3 inflammasome activation, BV2 cells were stimulated with LPS and then treated with ATP or nigericin. The secretion of IL-1ß was measured by ELISA. The expressions of NLRP3 inflammasome-related proteins and phospho IκBα/IκBα were analyzed using Western blotting. The production of intracellular and mitochondrial ROS was analyzed by flow cytometry. Results: Our results showed that punicalagin attenuated inflammation with reduction of pro-inflammatory mediators and cytokines including iNOS, COX-2, IL-1ß, and reduction of IL-6 led to inhibition of STAT3 phosphorylation by LPS-induced BV2 cells. Punicalagin also suppressed the ERK, JNK, and p38 phosphorylation, attenuated NF-κB activity, inhibited the activation of the NLRP3 inflammasome, and reduced the production of intracellular and mitochondrial ROS by LPS-induced BV2 cells. Conclusion: Our results demonstrated that punicalagin attenuated LPS-induced inflammation through suppressing the expression of iNOS and COX-2, inhibited the activation of MAPK/NF-κB signaling pathway and NLRP3 inflammasome, and reduced the production of ROS in microglia, suggesting that punicalagin might have the potential in treating neurodegenerative diseases.
ABSTRACT
PURPOSE: To describe a rare case of systemic paraquat poisoning presenting with peripheral ulcerative keratitis. METHODS: Case report and literature review. RESULTS: Two days after a mouthful of paraquat ingestion, a 48-year-old man presented with painful oral ulcers, abnormal liver functions, and acute kidney injury, followed by the development of crescent-shaped corneal erosions along the limbus and progressive visual impairment in both eyes. Paraquat-induced peripheral ulcerative keratitis was suspected and the patient was treated with intensive topical steroid along with systemic steroid. He recovered completely with good visual outcomes after treatment. CONCLUSIONS: Peripheral ulcerative keratitis is usually an immune-mediated ocular condition, and may be a complication associated with systemic paraquat poisoning. Proper diagnosis and adequate treatment are important for visual recovery.
Subject(s)
Corneal Ulcer/chemically induced , Herbicides/poisoning , Paraquat/poisoning , Acute Kidney Injury/chemically induced , Chemical and Drug Induced Liver Injury/etiology , Corneal Ulcer/diagnosis , Humans , Kidney Function Tests , Liver Diseases , Liver Function Tests , Male , Middle Aged , Oral Ulcer/chemically inducedABSTRACT
Purpose: To investigate the retinal sensitivity of highly myopic eyes with chorioretinal patchy atrophy (PA) using microperimetry. Methods: Fifty-two eyes of 32 patients with high myopia were prospectively included. Twenty-two eyes of 16 patients had PA lesions; eyes without PA were analyzed as controls. Testing points on microperimetry in eyes with PA were designated as 3 zones: zone 1 as the PA lesion including its borders; zone 2 including testing points adjoining PA; zone 3 including all other testing points. Results: In the PA group, the mean retinal sensitivity in zone 1 was 2.1 ± 2.8 dB, zone 2 = 8.3 ± 4.3 dB, and zone 3 = 9.4 ± 4.1 dB. Sensitivity in zone 1 was significantly reduced than zones 2 and 3 (P < 0.001). The mean retinal sensitivity in the PA group was lower than controls (6.5 ± 4.3 vs 13.9 ± 4.1 dB, P < 0.001), and combined zone 2 and 3 in the PA group also presented lower retinal sensitivity (8.8 ± 4.0 dB). Conclusions: Eyes with PA generate patchy scotoma in PA lesions and reduced retinal sensitivity in regions beyond atrophic lesion on microperimetry. The presence of PA may be an indicator to reflect both significantly anatomical and functional alterations on myopic macular degeneration.
Subject(s)
Macular Degeneration/pathology , Myopia/pathology , Scotoma/pathology , Adult , Aged , Aged, 80 and over , Analysis of Variance , Bruch Membrane/pathology , Female , Humans , Male , Middle Aged , Prospective Studies , Tomography, Optical CoherenceABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Ludwigia octovalvis (Jacq.) P.H. Raven (Onagraceae) extracts have historically been consumed as a healthful drink for treating various conditions, including edema, nephritis, hypotension and diabetes. AIM OF THE STUDY: We have previously shown that Ludwigia octovalvis extract (LOE) can significantly extend lifespan and improve age-related memory deficits in Drosophila melanogaster through activating AMP-activated protein kinase (AMPK). Since AMPK has become a critical target for treating diabetes, we herein investigate the anti-hyperglycemic potential of LOE. MATERIALS AND METHODS: Differentiated C2C12 muscle cells, HepG2 hepatocellular cells, streptozotocin (STZ)-induced diabetic mice and high fat diet (HFD)-induced diabetic mice were used to investigate the anti-hyperglycemic potential of LOE. The open field test and novel object recognition test were used to evaluate spontaneous motor activity and memory performance of HFD-induced diabetic mice. RESULTS: In differentiated C2C12 muscle cells and HepG2 hepatocellular cells, treatments with LOE and its active component (ß-sitosterol) induced significant AMPK phosphorylation. LOE also enhanced uptake of a fluorescent glucose derivative (2-NBDG) and inhibited glucose production in these cells. The beneficial effects of LOE were completely abolished when an AMPK inhibitor, dorsomorphin, was added to the culture system, suggesting that LOE requires AMPK activation for its action in vitro. In streptozotocin (STZ)-induced diabetic mice, we found that both LOE and ß-sitosterol induced an anti-hyperglycemic effect comparable to that of metformin, a drug that is commonly prescribed to treat diabetes. Moreover, LOE also improved glycemic control and memory performance of mice fed a HFD. CONCLUSIONS: These results indicate that LOE is a potent anti-diabetic intervention that may have potential for future clinical applications.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Hypoglycemic Agents/pharmacology , Onagraceae/chemistry , Plant Extracts/pharmacology , AMP-Activated Protein Kinases/metabolism , Animals , Blood Glucose/drug effects , Cell Line , Diabetes Mellitus, Experimental/physiopathology , Diet, High-Fat/adverse effects , Glucose/metabolism , Hep G2 Cells , Humans , Hypoglycemic Agents/isolation & purification , Male , Memory/drug effects , Mice , Mice, Inbred C57BL , StreptozocinABSTRACT
PURPOSE: The aim of this study was to evaluate a DNA dot hybridization assay (DHA) for assessing bacterial bioburden in orthokeratology lens (OK) storage cases. METHODS: Forty-one OK wearers participated in this study. The dot hybridization assay was used to assess the bacterial bioburden of OK after removal and 6-hour soaking in a storage case. Signals of the DHA were standardized after gray image transformation. The correlations between the hybridization intensities of three universal bacteria probes (BP1, BP2, and BP3) and bacterial bioburden determined by culture (colony forming units per milliliter) was analyzed by Pearson's correlation coefficient and receiver operating characteristic plots. In addition, three genus-specific probes for Pseudomonas, Acinetobacter, and Klebsiella were used to detect potentially hazardous bacterial contamination regardless of bacterial viability status. RESULTS: Among the three universal probes, there were good correlations between probe BP2 (r2 = 0.31, P = 9.5 × 10(-5)) and probe BP3 (r2 = 0.35, P = 3.1 × 10(-5)) with bacterial bioburden, but no correlation was found between probe BP1 and bacterial bioburden (r2 = 0.04, P = 0.11). In 41 samples, one was Pseudomonas-positive by both DHA and culture, while 10 were Pseudomonas-positive by DHA but negative by culture. Furthermore, nine samples tested positive for Acinetobacter (n = 7) and Klebsiella (n = 2) by DHA only. CONCLUSIONS: The dot hybridization assay provides a novel way to assess the bacterial bioburden of OK storage cases. Lens care quality can be assessed with universal bacteria probes, while potentially hazardous bacterial contamination can be traced with genus-specific probes.
Subject(s)
Bacteria/isolation & purification , Contact Lenses/microbiology , DNA, Bacterial/analysis , Equipment Contamination , Nucleic Acid Hybridization/methods , Orthokeratologic Procedures/instrumentation , Product Packaging , Adolescent , Bacteria/genetics , Bacterial Physiological Phenomena , Biofilms/growth & development , Child , Colony Count, Microbial , DNA Probes/chemistry , Female , Humans , Male , Polymerase Chain Reaction , Prospective StudiesABSTRACT
The rapidly increased availability of always-on broadband telecommunication environments and lower-cost vital signs monitoring devices bring the advantages of telemedicine directly into the patient's home. Hence, the control of access to remote medical servers' resources has become a crucial challenge. A secure authentication scheme between the medical server and remote users is therefore needed to safeguard data integrity, confidentiality and to ensure availability. Recently, many authentication schemes that use low-cost mobile devices have been proposed to meet these requirements. In contrast to previous schemes, Khan et al. proposed a dynamic ID-based remote user authentication scheme that reduces computational complexity and includes features such as a provision for the revocation of lost or stolen smart cards and a time expiry check for the authentication process. However, Khan et al.'s scheme has some security drawbacks. To remedy theses, this study proposes an enhanced authentication scheme that overcomes the weaknesses inherent in Khan et al.'s scheme and demonstrated this scheme is more secure and robust for use in a telecare medical information system.