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AIM: The influence of hypercholesterolemia on the development of apical periodontitis (AP) is inconclusive. Recent studies revealed that cholesterol metabolite 27-hydoxycholesterol (27HC) can affect cellular responses to bacterial infections and oestrogen status and raloxifene may influence its action. Herein, we aimed to examine the impact of 27HC on production of inflammatory mediators by macrophages and the regulatory function of raloxifene. The contribution of 27HC to AP development and the therapeutic effect of raloxifene were evaluated in a rat model. METHODS: Murine macrophages J774 cells were used. The expression of inducible nitric oxide synthase (iNOS) was examined by Western blot. The concentrations of C-C motif chemokine ligand (CCL) 2 and 27HC were assessed by enzyme-linked immunosorbent assay. Colorimetric assay was used to evaluate cholesterol levels. Experimental AP was induced in ovariectomized (OVX) or un-operated rats receiving high-fat/high-cholesterol diet (HFHCD) or normal diet (ND). Micro-computed tomography and immunohistochemistry were employed to evaluate disease severity and the therapeutic effect of raloxifene. RESULTS: Cholesterol enhanced 27HC production in macrophages. 27HC induced iNOS and CCL2 synthesis by macrophages and estradiol suppressed the responses. In our animal model of AP, HFHCD plus OVX significantly augmented serum and lesion tissue levels of 27HC (p < .05 versus the ND group). Lesion size, infiltration of CD68+ cells, and iNOS+ monocytes were increased in parallel with 27HC accumulation. Raloxifene inhibited pro-inflammatory effects of 27HC on macrophages and suppressed AP progression in HFHCD/OVX rats (p < .05 versus the vehicle control group). CONCLUSIONS: Our results suggested that 27HC contributes to AP aggravation associated with hypercholesterolemia. Oestrogen deficiency may both enhance 27HC production and exacerbate its downstream action.

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Objective: Sensorimotor performance is influential in Chinese handwriting, but few studies have examined the efficacy of sensorimotor-based interventions on Chinese handwriting among primary school students with poor handwriting performance. The study aims to evaluate a sensorimotor-based intervention to improve handwriting in the mainstream primary schools. Methods: This study adopted a two-group pretest-posttest design. An 8-session group-based sensorimotor intervention was delivered to school-aged children (mean age = 8.1, 68% male). Group A had 2 sessions every week, while Group B had 4 sessions every week. Analysis of variance with repeated measures was used to test the effects. Results: The intervention had a significant time effect (p < .05) in terms of improving handwriting process (d = 0.33-1.10), manual dexterity (d = 0.57), visual memory (d = 0.70), visual-spatial perception (d = 0.37), and motor and postural skills (d = 0.73). The effect sizes ranged from medium to large. For the handwriting process, time per character had a significant group × time interaction, with post hoc analysis showing that Group A had a significantly large effect (d = 1.89, p < .001) while Group B did not. Conclusions: The group-based sensorimotor intervention programme appeared to show improvements in students with fair skills in writing Chinese characters. It appears that the effect is better if the training sessions are spaced out in one month rather than intensively conducted within two weeks. It might be related to more involvement from parents, and students need more time for practice after the training sessions.

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AIM: To examine the role of palmitic acid in lipopolysaccharide (LPS)-stimulated chemotaxis of macrophages and the potential contribution of saturated fatty acid in signalling during the pathogenesis of apical periodontitis. METHODOLOGY: J774, a mouse macrophage cell line, was used in the experiments. After treatment with LPS, proteolytic maturation of sterol regulatory element-binding protein-1c (SREBP-1c) and expression of fatty acid synthase (FASN) were examined by Western analysis. Levels of palmitic acid were measured by reverse phase-high performance liquid chromatography-mass spectrometry. Knockdown of SREBP-1c and FASN was accomplished by small interfering RNA technology. Secretion of CC-chemokine ligand 2 (CCL2) and cellular chemotaxis were assessed by enzyme-linked immunosorbent assay and transwell migration assay, respectively. Sulfo-N-succinimidyl oleate (SSO) treatment was used to inhibit fatty acid signalling in vitro and also in a rat model of apical periodontitis. All data were first subjected to Levene's test. In vitro data were then analysed using ANOVA followed by Tukey's multiple comparison test. Data from animal experiments were analysed by independent t-tests. The significant level was set at 0.05. RESULTS: LPS stimulated proteolytic maturation of SREBP-1c and FASN expression in macrophages and significantly enhanced palmitic acid synthesis (P < 0.05). Knockdown of SREBP-1c attenuated LPS-enhanced FASN expression. Knockdown of FASN significantly suppressed LPS-enhanced palmitic acid synthesis (P < 0.05). LPS and exogenous palmitic acid significantly enhanced CCL2 secretion and macrophage chemotaxis (all P < 0.05). Inhibition of FASN expression significantly alleviated LPS-augmented CCL2 secretion (P < 0.05). SSO significantly suppressed CCL2 secretion and macrophage chemotaxis augmented by LPS and palmitic acid (all P < 0.05). In a rat model of induced apical periodontitis, SSO treatment significantly attenuated progression of apical periodontitis and macrophage recruitment (all P < 0.05). CONCLUSIONS: LPS/SREBP-1c/FASN/palmitic acid signalling contributed to tissue destruction caused by bacterial infection. Modulation of lipid metabolism and signalling may be helpful for the management of apical periodontitis.

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Lipopolisacáridos , Periodontitis Periapical , Animales , Ácidos Grasos , Macrófagos , Ratones , Ratas , Proteína 1 de Unión a los Elementos Reguladores de Esteroles

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AIM: To assess the connection between mitophagy and hypoxia-induced apoptosis in osteoblasts and whether simvastatin alleviates bone resorption in apical periodontitis through modulation of mitophagy-related apoptosis. METHODOLOGY: Hypoxia-induced generation of reactive oxygen species in mitochondria and changes in mitochondrial membrane potential were evaluated, respectively, by MitoSOX and JC-1 fluorescence dye signalling. Accumulation of mitophagy markers PTEN-induced putative kinase 1 (PINK1) and Parkin in mitochondria was examined by Western blotting and immunofluorescence microscopy. Osteoblast apoptosis was assessed by Western analysis of cleaved-poly (adenosine diphosphate ribose) polymerase (PARP). In a rat model of induced apical periodontitis, the therapeutic effect of simvastatin and its action on osteoblast mitophagy and apoptosis were examined. anova, Fisher's and Student's t-test were used for data analysis. RESULTS: Hypoxia-induced mitochondrial dysfunction and stimulated mitophagy in osteoblasts. Hypoxia also provoked apoptosis in osteoblasts and inhibition of mitophagy decreased hypoxia-augmented apoptotic activity. Simvastatin alleviated hypoxia-induced mitochondrial dysfunction, mitophagy and apoptosis. The protective action of simvastatin against apoptosis was related to its antimitophagy activity. Experiments in the rat model of induced apical periodontitis supported the laboratory findings. Simvastatin treatment mitigated periapical bone loss and reduced the activities of apoptosis and mitophagy in regional osteoblasts. CONCLUSIONS: The results suggest that modulation of osteoblast mitophagy may help diminish bone loss associated with inflammation and has potential as an auxiliary therapy for apical periodontitis.

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Resorción Ósea , Periodontitis Periapical , Animales , Apoptosis , Humanos , Mitofagia , Osteoblastos , Ratas , Simvastatina

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WW domain-containing oxidoreductase (WWOX), a putative tumour suppressor, is suggested to be involved in the hyperphosphorylation of Alzheimer's Tau. Tau is a microtubule-associated protein that has an important role in microtubule assembly and stability. Glycogen synthase kinase 3ß (GSK3ß) has a vital role in Tau hyperphosphorylation at its microtubule-binding domains. Hyperphosphorylated Tau has a low affinity for microtubules, thus disrupting microtubule stability. Bioinformatics analysis indicated that WWOX contains two potential GSK3ß-binding FXXXLI/VXRLE motifs. Immunofluorescence, immunoprecipitation and molecular modelling showed that WWOX interacts physically with GSK3ß. We demonstrated biochemically that WWOX can bind directly to GSK3ß through its short-chain alcohol dehydrogenase/reductase domain. Moreover, the overexpression of WWOX inhibited GSK3ß-stimulated S396 and S404 phosphorylation within the microtubule domains of Tau, indicating that WWOX is involved in regulating GSK3ß activity in cells. WWOX repressed GSK3ß activity, restored the microtubule assembly activity of Tau and promoted neurite outgrowth in SH-SY5Y cells. Conversely, RNAi-mediated knockdown of WWOX in retinoic acid (RA)-differentiated SH-SY5Y cells inhibited neurite outgrowth. These results suggest that WWOX is likely to be involved in regulating GSK3ß activity, reducing the level of phosphorylated Tau, and subsequently promoting neurite outgrowth during neuron differentiation. In summary, our data reveal a novel mechanism by which WWOX promotes neuronal differentiation in response to RA.

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Glucógeno Sintasa Quinasa 3/metabolismo , Neurogénesis/efectos de los fármacos , Neuronas/metabolismo , Oxidorreductasas/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Sitios de Unión , Línea Celular Tumoral , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Glucógeno Sintasa Quinasa 3/genética , Glucógeno Sintasa Quinasa 3 beta , Humanos , Microtúbulos/metabolismo , Simulación de Dinámica Molecular , Datos de Secuencia Molecular , Neuronas/citología , Oxidorreductasas/antagonistas & inhibidores , Oxidorreductasas/química , Fosforilación , Unión Proteica , Estructura Terciaria de Proteína , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Tretinoina/farmacología , Proteínas Supresoras de Tumor/antagonistas & inhibidores , Proteínas Supresoras de Tumor/química , Oxidorreductasa que Contiene Dominios WW , Proteínas tau/antagonistas & inhibidores , Proteínas tau/genética , Proteínas tau/metabolismo

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PURPOSE: To evaluate the efficacy, safety, predictability, and surgically induced astigmatism (SIA) of laser in situ keratomileusis (LASIK) for the correction of myopia and myopic astigmatism. SETTING: Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan. METHODS: This retrospective study comprised 69 eyes that had LASIK to correct myopia and 74 eyes that had LASIK to correct myopic astigmatism. The excimer laser keratectomy was performed using a Summit Apex Plus machine. Refraction, visual acuity, and computerized corneal videokeratography data from the preoperative and postoperative examinations were collected. The astigmatic change was calculated by the Alpins vector analysis method. RESULTS: The preoperative spherical equivalent at the glasses plane in the myopia and myopic astigmatism groups was -8.08 diopters (D) and -9.73 D, respectively. At 6 months, the spherical equivalent and residual corneal astigmatism were -0.25 D and 0.85 D, respectively, in the myopia group and -0.71 D and 0.82 D, respectively, in the myopic astigmatism group. In the myopia group, 88% of eyes were within +/-1.0 D of the intended myopia correction and in the myopic astigmatism group, 85% were within +/-1.0 D of the targeted spherical equivalent and 90% were within +/-1.0 D of the intended astigmatism correction. The uncorrected visual acuity was 20/40 or better in 94.1% of eyes in the myopia group and 92.5% of eyes in the myopic astigmatism group. The SIA magnitude was 0.66 D with the axis randomly distributed in the myopia group. The mean astigmatism correction index was 0.97, the mean magnitude of error was 0.13 D +/- 0.62 (SD), and the mean angle of error was -3.70 +/- 13.73 degrees in the myopic astigmatism group. CONCLUSION: Laser in situ keratomileusis had similar predictability, safety, and efficacy in the treatment of myopia and myopic astigmatism. The astigmatism correction was effective, but the results suggest that subjective astigmatism of less than 1.0 D need not be treated with the Summit Apex Plus laser.

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Astigmatismo/cirugía , Córnea/cirugía , Queratomileusis por Láser In Situ/métodos , Miopía/cirugía , Adulto , Astigmatismo/complicaciones , Topografía de la Córnea , Femenino , Humanos , Masculino , Persona de Mediana Edad , Miopía/complicaciones , Refracción Ocular , Estudios Retrospectivos , Seguridad , Resultado del Tratamiento , Agudeza Visual