RESUMO
Pterygium and subconjunctival hemorrhage are two common types of ocular surface diseases that can cause distress and anxiety in patients. In this study, 2855 ocular surface images were collected in four categories: normal ocular surface, subconjunctival hemorrhage, pterygium to be observed, and pterygium requiring surgery. We propose a diagnostic classification model for ocular surface diseases, dual-branch network reinforced by PFM block (DBPF-Net), which adopts the conformer model with two-branch architectural properties as the backbone of a four-way classification model for ocular surface diseases. In addition, we propose a block composed of a patch merging layer and a FReLU layer (PFM block) for extracting spatial structure features to further strengthen the feature extraction capability of the model. In practice, only the ocular surface images need to be input into the model to discriminate automatically between the disease categories. We also trained the VGG16, ResNet50, EfficientNetB7, and Conformer models, and evaluated and analyzed the results of all models on the test set. The main evaluation indicators were sensitivity, specificity, F1-score, area under the receiver operating characteristics curve (AUC), kappa coefficient, and accuracy. The accuracy and kappa coefficient of the proposed diagnostic model in several experiments were averaged at 0.9789 and 0.9681, respectively. The sensitivity, specificity, F1-score, and AUC were, respectively, 0.9723, 0.9836, 0.9688, and 0.9869 for diagnosing pterygium to be observed, and, respectively, 0.9210, 0.9905, 0.9292, and 0.9776 for diagnosing pterygium requiring surgery. The proposed method has high clinical reference value for recognizing these four types of ocular surface images.
RESUMO
Alkannin (ALK) has anti-inflammatory and anti-tumour activities. We tried to probe the underlying functions of ALK in oral squamous carcinoma (OSCC) cells growth, migration and invasion. OSCC cells viability was investigated after treatment with ALK. Then, BrdU, flow cytometry, Western blot and Transwell assays were executed to appraise proliferation, apoptosis, migration and invasion in OSCC cells with ALK stimulation. The biological functions of miR-9 were explored after miR-9 mimic/inhibitor transfection. The relevance of RECK and miR-9 was predicated by dual luciferase activity assay. JAK1/STAT3 and PI3K/AKT pathways were estimated by Western blot. Tumour formation in vivo was executed by xenograft tumours assay. We found that ALK restrained cell proliferation, facilitated apoptosis, repressed migration and invasion also interdicted JAK1/STAT3 and PI3K/AKT pathways in CAL-27 and SCC-9 cells. miR-9 expression was upgraded in OSCC tissues but decreased in OSCC cells along with ALK administration; meanwhile, overexpressed miR-9 inverted the influences of ALK in OSCC cells growth, migration and invasion. RECK was predicated as a novel target gene of miR-9, and overexpressed RECK hindered JAK1/STAT3 and PI3K/AKT pathways in OSCC cells. ALK prohibited tumour formation in vivo. In conclusion, ALK restrained OSCC cells growth, migration and invasion via adjusting miR-9/RECK axis. Highlights ALK restrains cell growth, migration and invasion in OSCC cells; miR-9 is enhanced in OSCC tissues but is repressed by ALK in OSCC cells; miR-9 inverts the repressive effect of ALK on CAL-27 and SCC-9 cells; RECK is a novel target of miR-9; ALK or RECK hinders JAK1/STAT3 and PI3K/AKT pathways in CAL-27 and SCC-9 cells; ALK prohibits tumour formation in vivo.
Assuntos
Antineoplásicos/farmacologia , Carcinoma de Células Escamosas/patologia , Movimento Celular/efeitos dos fármacos , Proteínas Ligadas por GPI/metabolismo , MicroRNAs/genética , Neoplasias Bucais/patologia , Naftoquinonas/farmacologia , Adulto , Idoso , Apoptose/efeitos dos fármacos , Carcinoma de Células Escamosas/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Proteínas Ligadas por GPI/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Janus Quinase 1/metabolismo , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/genética , Invasividade Neoplásica , Fator de Transcrição STAT3/metabolismoRESUMO
Glycyrrhizin (GL), a triterpenoid glycoside, serves important functions in various biological activities, including antiviral and antitumor immune responses. However, the anti-inflammatory effects of GL on Salmonella enterica serovar Typhimurium (ST)-induced injury in mice and the mechanisms underlying the protection of GL are poorly understood. Here, we investigated the effects of GL on host immune responses against ST infection in mice. A phenotypic analysis using hematoxylin and eosin (H&E) staining and transmission electron microscopy showed that GL relieved ST-induced weight loss and intestinal mucosal injury. A colonization assay showed that GL significantly reduced ST colonization in the ileum and colon and translocation to the liver and spleen. An antibacterial activity assay and real-time PCR revealed that GL had no direct inhibitory impact on ST growth or virulence gene expression. ELISA showed that GL pretreatment significantly decreased proinflammatory cytokine (IFN-γ, TNF-α, IL-6) secretion and increased anti-inflammatory cytokine (IL-10) secretion in the ileum, colon and serum of ST-infected mice. Moreover, flora analysis showed that GL reduced Akkermansia, Sutterella, Prevotella and Coprococcus but enriched Parabacteroides and Anaerotruncus in the cecum of ST-infected mice. These results suggest that GL promotes the secretion of immune factors and modulates intestinal flora to prevent further ST infection. We also analyzed the effect of GL on immunocytes and found that GL promoted the phenotypic and functional maturation of murine bone marrow-derived dendritic cells (BMDCs). Flow cytometry and western blotting demonstrated that NF-κB, ERK, and p38 MAPK were required for GL-induced BMDC maturation. The above findings indicate that GL attenuates ST infection by modulating immune function and intestinal flora. This study enriches our current knowledge of GL-mediated immunological function and provides a new perspective on the prevention of Salmonella infection in animals and humans.
Assuntos
Ácido Glicirrízico/farmacologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Interações Hospedeiro-Patógeno/imunologia , Infecções por Salmonella/imunologia , Infecções por Salmonella/microbiologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/fisiologia , Animais , Apresentação de Antígeno , Citocinas/biossíntese , Citocinas/sangue , Citotoxicidade Imunológica , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Feminino , Microbioma Gastrointestinal , Regulação Bacteriana da Expressão Gênica , Mediadores da Inflamação/sangue , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Infecções por Salmonella/metabolismo , Infecções por Salmonella/patologia , Salmonella typhimurium/patogenicidade , Transdução de Sinais , Virulência/genéticaRESUMO
OBJECTIVE: Salmonella enterica remains a major cause of food-borne disease in humans, and Salmonella Typhimurium (ST) contamination of poultry products is a worldwide problem. Since macrophages play an essential role in controlling Salmonella infection, the aim of this study was to evaluate the effect of glycyrrhizic acid (GA) on immune function of chicken HD11 macrophages. METHODS: Chicken HD11 macrophages were treated with GA (0, 12.5, 25, 50, 100, 200, 400, or 800 µg/ml) and lipopolysaccharide (LPS, 500 ng/ml) for 3, 6, 12, 24, or 48 h. Evaluated responses included phagocytosis, bacteria-killing, gene expression of cell surface molecules (cluster of differentiation 40 (CD40), CD80, CD83, and CD197) and antimicrobial effectors (inducible nitric oxide synthase (iNOS), NADPH oxidase-1 (NOX-1), interferon-γ (IFN-γ), LPS-induced tumor necrosis factor (TNF)-α factor (LITAF), interleukin-6 (IL-6), and IL-10), and production of nitric oxide (NO) and hydrogen peroxide (H2O2). RESULTS: GA increased the internalization of both fluorescein isothiocyanate (FITC)-dextran and ST by HD11 cells and markedly decreased the intracellular survival of ST. We found that the messenger RNA (mRNA) expression of cell surface molecules (CD40, CD80, CD83, and CD197) and cytokines (IFN-γ, IL-6, and IL-10) of HD11 cells was up-regulated following GA exposure. The expression of iNOS and NOX-1 was induced by GA and thereby the productions of NO and H2O2 in HD11 cells were enhanced. Notably, it was verified that nuclear factor-κB (NF-κB) and c-Jun N-terminal kinase (JNK) pathways were responsible for GA-induced synthesis of NO and IFN-γ gene expression. CONCLUSIONS: Taken together, these results suggested that GA exhibits a potent immune regulatory effect to activate chicken macrophages and enhances Salmonella-killing capacity.
Assuntos
Ácido Glicirrízico/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Salmonella/efeitos dos fármacos , Animais , Células Cultivadas , Galinhas , NF-kappa B/fisiologia , Fagocitose/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
The roots and rhizomes of Glycyrrhiza species (licorice) have been widely used as natural sweeteners and herbal medicines. The aim of this study is to investigate the effect of glycyrrhizic acid (GA) from licorice on macrophage polarization. Both phenotypic and functional activities of murine bone marrow-derived macrophages (BMDMs) treated by GA were assessed. Our results showed that GA obviously increased the cell surface expression of CD80, CD86, and MHCII molecules. Meanwhile, GA upregulated the expression of CCR7 and the production of TNF-α, IL-12, IL-6, and NO (the markers of classically activated (M1) macrophages), whereas it downregulated the expression of MR, Ym1, and Arg1 (the markers of alternatively activated (M2) macrophage). The functional tests showed that GA dramatically enhanced the uptake of FITC-dextran and E. coli K88 by BMDMs and decreased the intracellular survival of E. coli K88 and S. typhimurium. Moreover, we demonstrated that JNK and NF-κB activation are required for GA-induced NO and M1-related cytokines production, while ERK1/2 pathway exhibits a regulatory effect via induction of IL-10. Together, these findings indicated that GA promoted polarization of M1 macrophages and enhanced its phagocytosis and bactericidal capacity. The results expanded our knowledge about the role of GA in macrophage polarization.
Assuntos
Células da Medula Óssea/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Ácido Glicirrízico/farmacologia , MAP Quinase Quinase 4/fisiologia , Macrófagos/efeitos dos fármacos , NF-kappa B/fisiologia , Animais , Células da Medula Óssea/fisiologia , Ativação Enzimática , Macrófagos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico/biossíntese , Fagocitose/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
This study was conducted to investigate the effects of Bacillus subtilis B10 spores on the viability and biological functions of murine macrophage. RAW 264.7 cells were stimulated both with and without B. subtilis B10 spores for 12 h. Then cell viability was determined to evaluate the cytotoxic effect of B. subtilis B10 spores to the cells, and the activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH), the production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), and the secretion of inflammatory cytokines were measured to analyze the functions of macrophages. The results showed that B. subtilis B10 spores were not harmful to RAW 264.7 cells and they also strongly enhanced the activities of ACP and LDH (P < 0.01), remarkably increased NO and iNOS production (P < 0.01), and significantly stimulated the secretion of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-1 beta (IL-1ß), IL-6, IL-8 and IL-12 (P < 0.01) while they reduced anti-inflammatory cytokine IL-10 (P < 0.01). The outcomes suggest that B. subtilis B10 spores are not only safe for murine macrophages, but also can activate these cells and enhance their immune function. The above findings suggest that B. subtilis B10 spores are potentially probiotic.
Assuntos
Bacillus subtilis , Macrófagos/fisiologia , Fosfatase Ácida/análise , Animais , Sobrevivência Celular , Células Cultivadas , Indução Enzimática , Interferon gama/metabolismo , Interleucina-1beta/metabolismo , L-Lactato Desidrogenase/análise , Macrófagos/enzimologia , Camundongos , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/biossíntese , Probióticos , Esporos Bacterianos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
To investigate the immunomodulatory effects of Bacillus subtilis (B. subtilis) (natto) B4 spores on murine macrophage, RAW 264.7 cells were cultured alone or with B subtilis (natto) B4 spores at 37°C for 12 hrs, then both cells and culture supernatants were collected for analyses. Exposure of RAW 264.7 cells to B. subtilis (natto) B4 spores had no significant effects on macrophage viability and amounts of extracellular lactate dehydrogenase (LDH). However, it remarkably increased the activities of acid phosphatase (ACP), lactate dehydrogenase (LDH) and inducible nitric oxide synthase (iNOS) in cells and the amounts of nitric oxide (NO) and cytokines (tumor necrosis factor-alpha, interferon-gamma, interleukin [IL]-1 beta, IL-6, IL-12, IL-10 and macrophage inflammatory protein-2) in culture supernatants. These results demonstrate that B. subtilis (natto) B4 spores are harmless to murine macrophages and can stimulate their activation through up-regulation of ACP and LDH activities and enhance their immune function by increasing iNOS activity and stimulating NO and cytokine production. The above findings suggest that B. subtilis (natto) B4 spores have immunomodulatory effects on macrophages.