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Porphyromonas gingivalis (P. gingivalis) is one of the major pathogens causing periodontitis and apical periodontitis (AP). Long noncoding RNA (lncRNA) can regulate cellular mineralization and inflammatory diseases. The aim of this study was to investigate the role and mechanism of lncRNA in P. gingivalis-stimulated cementoblast mineralization. In vivo, C57BL/6 mice were divided into the healthy, the AP, and AP + P. gingivalis groups (n = six mice per group). Micro computed tomography, immunohistochemistry staining, and fluorescence in situ hybridization were used to observe periapical tissue. In vitro, cementoblasts were treated with osteogenic medium or P. gingivalis. Pluripotency associated transcript 3 (Platr3), interleukin 1 beta (IL1B), and osteogenic markers were analyzed by quantitative real-time polymerase chain reaction and western blot. RNA pull-down and RNA immunoprecipitation assays were used to detect proteins that bind to Platr3. RNA sequencing was performed in Platr3-silenced cementoblasts. In vivo, P. gingivalis promoted periapical tissue destruction and IL1B expression, but inhibited Platr3 expression. In vitro, P. gingivalis facilitated IL1B expression (P < 0.001), whereas suppressed the expression of Platr3 (P < 0.001) and osteogenic markers (P < 0.01 or 0.001). In contrast, Platr3 overexpression alleviated the repressive effect of P. gingivalis on cementoblast mineralization (P < 0.01 or 0.001). Furthermore, Platr3 bound to nudix hydrolase 21 (NUDT21) and regulated the nuclear factor-κB (NF-κB) signaling pathway. Knocking down NUDT21 suppressed osteogenic marker expression and activated the above signaling pathway. Collectively, the results elucidated that Platr3 mediated P. gingivalis-suppressed cementoblast mineralization through the NF-κB signaling pathway by binding to NUDT21.
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Oral diseases are among the most common diseases worldwide and are associated with systemic illnesses, and the rising occurrence of oral diseases significantly impacts the quality of life for many individuals. It is crucial to detect and treat these conditions early to prevent them from advancing. DNA methylation is a fundamental epigenetic process that contributes to a variety of diseases including various oral diseases. Taking advantage of its reversibility, DNA methylation becomes a viable therapeutic target by regulating various cellular processes. Understanding the potential role of this DNA alteration in oral diseases can provide significant advances and more opportunities for diagnosis and therapy. This article will review the biology of DNA methylation, and then mainly discuss the key findings on DNA methylation in oral cancer, periodontitis, endodontic disease, oral mucosal disease, and clefts of the lip and/or palate in the background of studies on global DNA methylation and gene-specific DNA methylation.
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Metilación de ADN , Epigénesis Genética , Enfermedades de la Boca , Humanos , Enfermedades de la Boca/genética , Enfermedades de la Boca/metabolismo , AnimalesRESUMEN
OBJECTIVE: To evaluate the surface electromyography (sEMG) of pelvic floor muscles (PFMs), compare between vaginal birth and cesarean section and correlate with maternity and obstetrics characteristics in primiparous 6-8 weeks postpartum. METHODS: PFMs surface electromyography screening data of primiparous postpartum women in our hospital at 6-8 weeks postpartum from 2018 to 2021 were selected and analyzed. The study collected data on delivery activities of 543 postpartum women totally. RESULTS: In general, the abnormal incidence of pelvic floor electromyography in postpartum women mainly occurred in slow muscle (type I fiber) stage and endurance testing stage. Compared to vaginal birth postpartum women, the incidence of abnormal pelvic floor electromyography in cesarean section postpartum women is lower. There were statistical differences in measurement values of pelvic floor electromyography in several different stages between cesarean section and vaginal birth (P < 0.005). Regarding the influence on pelvic floor electromyography, there were more influencing factors on vaginal birth postpartum women including age, height, weight, weight gain during pregnancy, gestational week, and first and second stage of labor than on cesarean section postpartum women whose influencing factors included age, weight gain during pregnancy, and newborn weight. CONCLUSION: Effects on surface electromyography (sEMG) of pelvic floor muscles (PFMs) at 6-8 weeks postpartum differed based on the different modes of delivery. The high-risk obstetric factors closely related to abnormal surface electromyography (sEMG) of pelvic floor muscles (PFMs) were maternal age, height, weight, and second stage of labor.
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Cesárea , Diafragma Pélvico , Embarazo , Recién Nacido , Femenino , Humanos , Estudios Transversales , Electromiografía , Periodo Posparto , Aumento de PesoRESUMEN
BACKGROUND: Cementoblasts on the tooth-root surface are responsible for cementum formation (cementogenesis) and sensitive to Porphyromonas gingivalis stimulation. We have previously proved transcription factor CXXC-type zinc finger protein 5 (CXXC5) participates in cementogenesis. Here, we aimed to elucidate the mechanism in which CXXC5 regulates P. gingivalis-inhibited cementogenesis from the perspective of mitochondrial biogenesis. METHODS: In vivo, periapical lesions were induced in mouse mandibular first molars by pulp exposure, and P. gingivalis was applied into the root canals. In vitro, a cementoblast cell line (OCCM-30) was induced cementogenesis and submitted for RNA sequencing. These cells were co-cultured with P. gingivalis and examined for osteogenic ability and mitochondrial biogenesis. Cells with stable CXXC5 overexpression were constructed by lentivirus transduction, and PGC-1α (central inducer of mitochondrial biogenesis) was down-regulated by siRNA transfection. RESULTS: Periapical lesions were enlarged, and PGC-1α expression was reduced by P. gingivalis treatment. Upon apical inflammation, Cxxc5 expression decreased with Il-6 upregulation. RNA sequencing showed enhanced expression of osteogenic markers, Cxxc5, and mitochondrial biogenesis markers during cementogenesis. P. gingivalis suppressed osteogenic capacities, mitochondrial biogenesis markers, mitochondrial (mt)DNA copy number, and cellular ATP content of cementoblasts, whereas CXXC5 overexpression rescued these effects. PGC-1α knockdown dramatically impaired cementoblast differentiation, confirming the role of mitochondrial biogenesis on cementogenesis. CONCLUSIONS: CXXC5 is a P. gingivalis-sensitive transcription factor that positively regulates cementogenesis by influencing PGC-1α-dependent mitochondrial biogenesis. Video Abstract.
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Cementogénesis , Mitocondrias , Biogénesis de Organelos , Animales , Ratones , Línea Celular , Cementogénesis/genética , Cementogénesis/fisiología , ADN Mitocondrial/metabolismo , Proteínas de Unión al ADN/metabolismo , Regulación de la Expresión Génica , Factores de Transcripción/metabolismo , Mitocondrias/metabolismoRESUMEN
Cementum, a constituent part of periodontal tissues, has important adaptive and reparative functions. It serves to attach the tooth to alveolar bone and acts as a barrier delimit epithelial growth and bacteria evasion. A dynamic and highly responsive cementum is essential for maintaining occlusal relationships and the integrity of the root surface. It is a thin layer of mineralized tissue mainly produced by cementoblasts. Cementoblasts are osteoblast-like cells essential for the restoration of periodontal tissues. In recent years, glucose metabolism has been found to be critical in bone remodeling and osteoblast differentiation. However, the glucose metabolism of cementoblasts remains incompletely understood. First, immunohistochemistry staining and in vivo tracing with 18 F-fluorodeoxyglucose (18 F-FDG) revealed significantly higher glucose metabolism in cementum formation. To test the bioenergetic pathways of cementoblast differentiation, we compared the bioenergetic profiles of mineralized and unmineralized cementoblasts. As a result, we observed a significant increase in the consumption of glucose and production of lactate, coupled with the higher expression of glycolysis-related genes. However, the expression of oxidative phosphorylation-related genes was downregulated. The verified results were consistent with the RNA sequencing results. Likewise, targeted energy metabolomics shows that the levels of glycolytic metabolites were significantly higher in the mineralized cementoblasts. Seahorse assays identified an increase in glycolytic flux and reduced oxygen consumption during cementoblast mineralization. Apart from that, we also found that lactate dehydrogenase A (LDHA), a key glycolysis enzyme, positively regulates the mineralization of cementoblasts. In summary, cementoblasts mainly utilized glycolysis rather than oxidative phosphorylation during the mineralization process.
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Cemento Dental , Ácido Láctico , Diferenciación Celular , Inmunohistoquímica , GlucosaRESUMEN
Objectives: Casein kinase 2 interacting protein-1 (CKIP-1) is a versatile player involved in various biological processes. However, whether CKIP-1 mediates the osteogenic/cementogenic differentiation of periodontal ligament cells (PDLCs) under Porphyromonas gingivalis (Pg) stimulation remains unknown. Material and Methods: The effect of Pg on PDLC differentiation was first verified. CKIP-1 expression in Pg-infected PDLCs or in PDL of apical periodontitis (AP) mice was detected. The changes of CKIP-1 during PDLC differentiation was also determined. PDLC differentiation capacity in CKIP-1 knockout (KO) mice and CKIP-1-silenced PDLCs with or without Pg stimulation were further studied. Inhibitor was finally applied to verify the involvement of p38 signaling pathway in PDLC differentiation. Results: The suppression effect of Pg on PDLC differentiation was demonstrated. CKIP-1 increased in the PDL of AP mice and Pg-induced PDLCs, and decreased gradually during PDLC differentiation. Increased OSX and RUNX2 expression in PDL were observed in CKIP-1 KO mice. Also, CKIP-1 silencing facilitated and rescued Pg-inhibited PDLC differentiation. Inhibitor for p38 signaling pathway blocked CKIP-1 silencing-facilitated PDLC differentiation. Conclusions: CKIP-1 mediated the osteogenic/cementogenic differentiation of PDLCs partially through p38 signaling pathway, which may provide evidence for the regeneration of periodontal hard tissues damaged by Pg.
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As a chronic inflammatory disease, periodontitis involves many biological processes including autophagy. At the same time, casein kinase 2 interacting protein-1 (CKIP-1) was reported to play a role in regulation of inflammation. But whether CKIP-1 and autophagy interact in periodontitis remains unclear. In this paper, our research team verified the levels of CKIP-1 expression and autophagy increase in the periodontal tissues of a ligature-induced periodontitis mouse model. And this result was also confirmed in Porphyromonas gingivalis (Pg)-induced human gingival fibroblasts (HGF) and human periodontal ligament cells (PDLC). We also showed the autophagy level in periodontal tissues is higher in Ckip-1 knockout (KO) mice than wild type (WT). At the same time, CKIP-1 knockdown lentivirus was used in PDLC and HGF, and it was found that silencing CKIP-1 significantly activated autophagy. Unfortunately, the regulatory role of autophagy in periodontitis is still unclear. Then, the autophagy agonist Rapamycin and inhibitor 3-MA were used in a periodontitis mouse model to investigate periodontal tissue destruction. We found the inflammation in periodontal tissue was reduced when autophagy activated. All these conclusions have been verified both in vivo and in vitro experiments. Finally, our research proved that silencing CKIP-1 reduces the expression of inflammatory cytokines in Pg-induced PDLC and HGF by regulating autophagy. Overall, a new role for CKIP-1 in regulating periodontal tissue inflammation was demonstrated in our study, and it is possible to treat periodontitis by targeting the CKIP-1 gene.
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Inflamación , Periodontitis , Ratones , Animales , Humanos , Inflamación/metabolismo , Periodontitis/metabolismo , Encía/metabolismo , Citocinas/metabolismo , Porphyromonas gingivalis/metabolismo , Autofagia , Proteínas Portadoras/metabolismoRESUMEN
Porphyromonas gingivalis is involved in the pathogenesis of multiple polymicrobial biofilm-induced inflammatory diseases, including apical periodontitis, and it triggers pyroptosis accompanied by robust inflammatory responses. Tet methylcytosine dioxygenase 1 (TET1), an epigenetic modifier enzyme, has been is correlated with inflammation, though an association of TET1 and P. gingivalis-related pyroptosis in cementoblasts and the molecular mechanisms has not been shown. Our study here demonstrated that P. gingivalis downregulated Tet1 expression and elicited CASP11- and GSDMD-dependent pyroptosis. Additionally, Tet1 mRNA silencing in cementoblasts appeared to result in a more severe pyroptotic phenotype, where levels of CASP11 and GSDMD cleavage, lactate dehydrogenase release, and IL-1ß and IL-18 production were significantly increased. Moreover, Tet1 overexpression resulted in blockade of pyroptosis activation accompanied by inflammation moderation. Further analyses revealed that TET1 modulated glycolysis, confirmed by the application of the specific inhibitor 2-deoxy-d-glucose (2-DG). The pyroptosis phenotype enhanced by Tet1 silencing was moderated by 2-DG upon P. gingivalis invasion. Taken together, these data show the effects and underlying mechanisms of TET1 on pyroptosis and inflammatory phenotype induced by P. gingivalis in cementoblasts, and provides insight into the involvement of P. gingivalis in apical periodontitis and, possibly, other inflammatory diseases.
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Dioxigenasas , Periodontitis Periapical , Humanos , Piroptosis , Porphyromonas gingivalis/metabolismo , Cemento Dental/metabolismo , Inflamación/metabolismo , Glucólisis , Dioxigenasas/metabolismo , Oxigenasas de Función Mixta/metabolismo , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismoRESUMEN
Uterus transplantation has become an option for women suffering from some form of infertility. Current review discusses key physiological functions of the endometrium requiring the transition of tissue cells between the mesenchyme and epithelial cell phenotype, a process known as epithelial-mesenchymal transition (EMT). Estrogen and EMT play a key role in the pathogenesis and treatment of intrauterine adhesion and endometriosis. There is also a close regulatory relationship between estrogen and EMT, and investigation of this relationship is of great significance for the treatment of endometrial disorders. The present review discusses the effects of estrogen on endometrial dysfunction, with a focus on the relationship between estrogen and EMT in endometrial disorders, taking into consideration the mechanisms by which receptors that regulate their functions and proteins that regulate their local biological functions interact with the factors involved in EMT. In addition, the review summarizes emerging drugs targeting receptors or proteins and provides information on the direction of new therapies for endometrial disorders.
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Endometriosis , Humanos , Femenino , Endometriosis/genética , Endometriosis/metabolismo , Endometriosis/patología , Transición Epitelial-Mesenquimal , Estrógenos/uso terapéutico , Estrógenos/metabolismo , Estrógenos/farmacología , Endometrio/metabolismo , Endometrio/patología , ÚteroRESUMEN
BACKGROUND: Cementum regeneration was regarded as the critical goal for periodontal regeneration, and M2 macrophage-based therapy was expected to be a promising strategy. However, little is known about the effects of M2 macrophages on cementoblast mineralization and tropism, especially under inflammation. Here we investigated for the first time the crosstalk between M2 macrophages and Porphyromonas gingivalis (Pg)-stimulated cementoblasts. METHODS: M2 macrophages were induced with interleukin (IL)-4, and identified. CC-chemokine ligand 2 (CCL2) expression and secretion of inflammatory cementoblasts were detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), western blotting (WB), immunohistochemistry for apical periodontitis (AP) mice, and by enzyme-linked immunosorbent assay. Crystal violet staining was used to observe macrophage migration. Conditional medium (CM) and transwell coculture methods were applied to evaluate the effects of M2 macrophages on cementum mineralization with or without Pg, and to explore the mechanism. Mineralization-related markers and pathway-related proteins were measured by RT-qPCR and WB. RESULTS: M2 macrophages were identified successfully. We found an increase of CCL2 in cementoblasts and their supernatant. Also, higher CCL2 in cementoblasts was observed in the AP model. Superior recruitment of M2 macrophages to supernatant from Pg-stimulated cementoblasts or CCL2-containing medium was verified. Moreover, CM2 and Trans-M2 showed better mineralization-accelerating and rescuing effects when compared to their controls, and application of p38 inhibitor partially blocked the promotion. CONCLUSIONS: Our study demonstrated the inflammation-targeting and mineralization-promoting effects of M2 macrophages on cementoblasts, which may provide evidence for M2 macrophage-based cementum regeneration.
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Cemento Dental , Macrófagos , Ratones , Animales , Cemento Dental/metabolismo , Macrófagos/metabolismo , Movimiento Celular , InflamaciónRESUMEN
Objective: To investigate the clinical effect of Multi-focused (MF) laser in the treatment of vulvar lichen sclerosus (VLS). Methods: In this single-center, randomized controlled trial, we compared the effect of fractionated MF laser with other treatments on patients with biopsy-proven VLS. Patients with VLS were enrolled in this study and randomly divided into three groups. Patients in the experimental group were treated with a CO2 laser, control group 1 was treated with radiofrequency, and control group 2 was treated topically with glucocorticoids and soaking with Chinese patent medicine. The pruritus degree, skin elasticity, skin color, lesion scope, and total score were compared before treatment, at one month after treatment, and three months after treatment. Results: One month after treatment, the pruritus degree, skin elasticity, skin color, lesion scope, and total score decreased in the experimental group, and the differences were statistically significant (P < 0.05). In control group 1, the differences in pruritus degree, skin color, and total score were statistically significant (P < 0.05), but the differences in skin elasticity and lesion scope were not statistically significant (P > 0.05). In control group 2, the differences in pruritus degree and total score were statistically significant (P < 0.05), but the differences in skin elasticity, skin color, and lesion scope were not statistically significant (P > 0.05). At one month after the end of treatment, the differences in pruritus degree, skin elasticity, skin color, lesion scope, and total score among the three groups were not statistically significant. At three months after the end of treatment, the differences in the scores of the five indicators were statistically significant. Conclusion: For the three treatment methods for VLS, topical corticosteroids + traditional Chinese medicine can quickly relieve itching symptoms in patients, but it cannot significantly improve skin elasticity, skin color, and lesion scope, and VLS easily relapses after treatment. Radiofrequency can improve itching symptoms and skin color but has poor effects on the change of skin elasticity and lesion scope. Multi-focused laser treatment can alleviate the degree of pruritus, improve skin color and elasticity, and narrow the lesion scope, and VLS will not relapse within three months after treatment.
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Context: Early diagnosis and early treatment of cornual pregnancy are very important. Conservative treatment before rupture can greatly reduce the patient's trauma. It's very important to choose a treatment method for cornual pregnancy with a high level of effectiveness, few adverse reactions, and no effects on fertility. Objective: The study intended to compare the clinical efficacy of different treatments for unruptured cornual pregnancy to find a safe, effective, minimally invasive treatment for unruptured cornual pregnancy that has few side effects and doesn't affect fertility. Design: The research team retrospectively collected the clinical data of patients to analyze the benefits of treatments for cornual pregnancy. Setting: The study took place in the Department of Obstetrics and Gynecology at the Wuhan Third Hospital in Wuhan, Hubei Province, China. Participants: Participants were 61 patients with an unruptured cornual pregnancy who had been admitted to the hospital between September 2002 and May 2012. Intervention: Participants were divided into four groups according to the treatment they received: (1) 20 patients who had been orally administered mifepristone combined with misoprostol and received uterine curettage were included in the drug abortion + curettage group (D group); (2) 16 patients who had received ultrasound-guided uterine aspiration were included in the uterine aspiration group (U group); (3) 15 patients who had received methotrexate (MTX) chemotherapy were included in the chemotherapy group (C group); and (4) 10 patients who had received ultrasound-guided hysteroscope operation were included in the hysteroscope operation group (H group). Outcome Measures: Adverse reactions and the decrease in participants' blood ß-HCG were recorded in detail. The participants were followed up for two months. Results: Of the 61 participants, 12 underwent surgery after failed conservative treatment, one in the D group, four in the U group, three in the C group, and four in the H group. No significant difference existed in the baseline data among the four groups. The decline rates of ß-HCG at seven days after treatment and the treatment success rates of participants in the D group were significantly higher than those in the U group, the C group, and the H group (all P < .05). The time at which the ß-HCG turned negative and the average hospital stays weren't significantly different among the four groups. Conclusions: The current study found that oral administration of mifepristone, combined with misoprostol, plus uterine curettage was superior to the other three methods in treatment of unruptured cornual pregnancy. The drug abortion + curettage treatment was found to be a safe, effective, minimally invasive treatment for unruptured cornual pregnancy, which has few side effects and doesn't affect fertility.
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Misoprostol , Embarazo Cornual , Tratamiento Conservador , Femenino , Humanos , Mifepristona/uso terapéutico , Embarazo , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
BACKGROUND: Spermatogenesis arrest and spermatogenic cell apoptosis occur in the testes of heat-stressed mice. Although heat stress-induced spermatogenic cell apoptosis is due to the decreased expression of cold-inducible RNA-binding protein (CIRBP), it remains unclear whether spermatogenesis arrest is also affected by CIRBP. Additionally, the specific mechanism by which CIRBP regulates spermatogenic cell apoptosis or inhibits spermatogenesis remains to be elucidated. OBJECTIVES: To investigate the mechanism by which CIRBP contributes to heat stress-induced testicular spermatogenesis arrest. MATERIALS AND METHODS: Target mRNAs downstream of CIRBP in testicular tissue of BALB/c mice, exposed or not to heat stress, were sequenced. Sequencing data were subjected to bioinformatics analysis to identify key mRNAs and pathways associated with heat stress-induced spermatogenic damage. The link between CIRBP and its target mRNA Ccnb1 (cyclin B1) was verified by western blotting, flow cytometry, and RNA pulldown assays, and the ability of CIRBP to inhibit germ cell cycle arrest by regulating cyclin B1 expression was investigated in a mouse spermatocyte cell line (GC-2spd). RESULTS: Changes in mRNA expression downstream of CIRBP were mainly associated with the cell cycle and RNA binding, transport and splicing. Cyclin B1 was found to regulate the G2/M transition during the first meiotic division of spermatogenic cells. Further, CIRBP was shown to bind directly to the 3'-untranslated region of Ccnb1 mRNA and was associated with cyclin B1-induced inhibition of spermatogenesis arrest. DISCUSSION AND CONCLUSION: In conclusion, our results provide strong evidence that CIRBP may exert its key function in heat stress-induced testicular spermatogenic cell injury partly by regulating the expression of Ccnb1, the product of which inhibits spermatogenesis arrest.
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Puntos de Control del Ciclo Celular/genética , Ciclina B1/metabolismo , Respuesta al Choque Térmico/genética , Proteínas de Unión al ARN/metabolismo , Espermatogénesis/genética , Animales , Apoptosis/genética , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/metabolismo , Testículo/metabolismoRESUMEN
OBJECTIVE: To identify the key genes involved in prostate cancer and their regulatory network. METHODS: The dataset of mRNA/miRNA transcriptome sequencing was downloaded from The Cancer Genome Atlas/the Gene Expression Omnibus database for analysis. The "edgeR" package in the R environment was used to normalize and analyze differentially expressed genes (DEGs) and miRNAs (DEmiRNAs). First, the PANTHER online tool was used to analyze the function enrichment of DEGs. Next, a protein-protein interaction (PPI) network was constructed using STRING and Cytoscape tools. Finally, miRNA-gene regulatory networks were constructed using the miRTarBase. RESULTS: We identified 4339 important DEGs, of which 2145 were upregulated (Up-DEGs) and 2194 were downregulated (Down-DEGs). Functional enrichment analysis showed that the Up-DEGs were related to the immune system and the cell cycle in prostate cancer, whereas the Down-DEGs were related to the nucleic acid metabolic process and metabolism pathways. Twelve core protein clusters were found in the PPI network. Further, the constructed miRNA-gene interaction network showed that 11 downregulated miRNAs regulated 16 Up-DEGs and 22 upregulated miRNAs regulated 22 Down-DEGs. CONCLUSION: We identified 4339 genes and 70 miRNAs that may be involved in immune response, cell cycle, and other key pathways of the prostate cancer regulatory network. Genes such as BUB1B, ANX1A1, F5, HTR4, and MUC4 can be used as biomarkers to assist in the diagnosis and prognosis of prostate cancer.
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Biología Computacional/métodos , Regulación Neoplásica de la Expresión Génica , Redes Reguladoras de Genes , Neoplasias de la Próstata/genética , Humanos , MasculinoRESUMEN
PURPOSE: The toxicity of copper nanoparticle (CuNP) exposure in the ovaries has attracted attention recently, but the precise molecular mechanism involved requires further investigation. We investigated the cytotoxicity of CuNPs in ovarian granulosa cells and the protective effect of heme oxygenase 1 (HO-1) against CuNP-induced damage. METHODS: Human ovarian granulosa cells (COV434) were treated with CuNPs, and cytotoxicity was evaluated using Cell Counting Kit-8 and flow cytometry assays. Oxidative stress was identified using biochemical markers of oxidation and anti-oxidation. The protein levels of mitogen-activated protein kinase 14 (MAPK14), phospho-MAPK14, nuclear factor erythroid 2-related factor 2 (Nrf2), and HO-1 were measured by immunoblotting. Subsequently, for oxidative stress parameter detection, the cells were pre-treated with hemin to induce HO-1 expression prior to CuNP treatment. RESULTS: Exposure to CuNPs decreased cell viability and the mitochondrial membrane potential, increased the apoptosis rate, and induced oxidative stress. Furthermore, hemin pretreatment induced HO-1 expression in cells, which partially reduced the accumulation of reactive oxygen species induced by CuNPs and increased the levels of antioxidant enzymes. CONCLUSION: CuNPs exert cytotoxic effects on human ovarian granulosa cells by inducing oxidative stress, and may induce HO-1 expression via the MAPK14-Nrf2 signaling pathway. Moreover, HO-1 protects against oxidative stress induced by CuNPs.
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Cobre/toxicidad , Hemo-Oxigenasa 1/farmacocinética , Nanopartículas del Metal/toxicidad , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Hemo-Oxigenasa 1/metabolismo , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Proteínas de Neoplasias/metabolismo , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND: MicroRNAs play a crucial role in the regulation of spermatogenesis. For example, miR-128-3p expression is known to decrease significantly after testicular hyperthermia, but the regulatory effect of this change on the spermatogenesis damage caused by heat stress remains unclear. OBJECTIVES: This study aimed to verify whether the target gene of miR-128-3p is MAPK14, which affects spermatogenic cell proliferation and apoptosis under testicular hyperthermia. MATERIALS AND METHODS: Mouse testis and GC2 spermatocyte cell line heat stress models were established. miR-128-3p expression before and after heat stress was analyzed by reverse transcription polymerase chain reaction. MAPK14 and p-MAPK14 expression was detected by Western blot, and cell apoptosis was analyzed by Annexin V-FITC/PI. Subsequently, miR-128-3p inhibitors and mimics were used to interfere with spermatocytes before and after heat stress, respectively, for correlation detection. RESULTS: Compared with the control group, the heat stress group showed decreased miR-128-3p expression, increased p-MAPK14 expression, and decreased cell proliferation activity. In the GC2-spd cell line in vitro, miR-128-3p inhibitors were found to upregulate p-MAPK14 expression, reduce cell proliferation activity, and increase apoptosis, consistent with the results obtained in the heat treatment alone. Furthermore, miR-128-3p mimics transfected in the GC2 cells after heat stress reduced p-MAPK14 expression, alleviated the decrease in cell proliferation, and decreased the apoptosis level. CONCLUSIONS: The downregulation of miR-128-3p expression plays an important role in spermatogenesis damages after testicular hyperthermia, which is probably attributable to the activation of the MAPK signaling pathway. Downregulated miR-128-3p expression induces the apoptosis and inhibits the proliferation of spermatogenic cells by promoting MAPK14 phosphorylation.
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Apoptosis/genética , MicroARNs/fisiología , Espermatocitos/fisiología , Testículo/metabolismo , Animales , Línea Celular , Activación Enzimática/genética , Regulación del Desarrollo de la Expresión Génica , Respuesta al Choque Térmico , Masculino , Ratones , Ratones Endogámicos ICR , Proteína Quinasa 14 Activada por Mitógenos , Espermatocitos/enzimología , Espermatogénesis/genética , Testículo/citología , Testículo/enzimologíaRESUMEN
BACKGROUND Hyperuricemia has a pathogenic role in the development of hypertension and other cardiovascular diseases (CVD). Uric acid has been reported to activate Nod-like receptor protein 3 (NLRP3)-inflammasome and alter vascular smooth muscle cells (VSMC). However, the potential mechanisms underlying this association are still not understood. The aim of this study was to investigate the role and potential mechanisms of uric acid in proliferation of VSMC. MATERIAL AND METHODS Cell Counting Kit-8 (CCK-8) proliferation assay and colony formation assay were performed to determine the proliferative ability of VSMC under uric acid stimulation. Immunofluorescence microscopy was carried out to determine the expression of Alpha-smooth muscle actin (alpha-SMA). In addition, real-time PCR and Western blot were used to detect the expression of NLRP3-inflammasome, and ELISA was performed to measure the levels of IL-18 and IL-1ß. RESULTS The results showed that uric acid increases the proliferation of VSMC and induces alpha-SMA accumulation. We also found that uric acid increases the level of NLRP3 and induces NLRP3-inflammasome activation. The expressions of uric acid-induced inflammatory markers IL-1ß and IL-18 were decreased by the inhibitor MCC950. CONCLUSIONS Our findings revealed that uric acid induces inflammation through NLRP3-inflammasome-mediated VSMC proliferation. NLRP3 may be a new therapeutic target for hypertension.
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Miocitos del Músculo Liso/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ácido Úrico/metabolismo , Actinas/metabolismo , Animales , Enfermedades Cardiovasculares/metabolismo , Proteínas Portadoras/metabolismo , Línea Celular , Proliferación Celular/efectos de los fármacos , China , Humanos , Hipertensión/tratamiento farmacológico , Hiperuricemia/tratamiento farmacológico , Inflamasomas/metabolismo , Inflamación/patología , Interleucina-18/metabolismo , Músculo Liso Vascular/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Cold-inducible RNA-binding protein (CIRBP) is reduced by scrotal hyperthermia in cryptorchidism, varicocoele and heat treatment, but there is no direct evidence clarifying the relationship between CIRBP and spermatogenesis. The aim of this study was to investigate the expression of CIRBP in GC2-spd cells (a mouse spermatocyte cell line) before and after heat treatment, and to determine the effects of the downregulation or overexpression of CIRBP on spermatocyte cell proliferation and apoptosis. GC2-spd cells overexpressing CIRBP and GC2-spd cells in CIRBP was knocked down were constructed to investigate the function of CIRBP in cell proliferation and apoptosis using a cell counting kit-8 and flow cytometry respectively. In addition, proliferation and apoptosis were evaluated in GC2-spd cells that had been heated for 30 or 60min, and were analysed 12, 24, and 48h after heat treatment. Heat treatment clearly suppressed the proliferation of GC2-spd cells, and upregulation of CIRBP expression in GC2-spd cells promoted cell proliferation and decreased apoptosis before and after heat stress; in contrast, downregulation of CIRBP expression inhibited cell proliferation and increased apoptosis. These findings suggest that CIRBP exerts a protective effect against spermatogenic injury caused by heat stress.
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Apoptosis/fisiología , Proliferación Celular/fisiología , Respuesta al Choque Térmico/fisiología , Proteínas de Unión al ARN/metabolismo , Espermatocitos/citología , Animales , Línea Celular , Regulación hacia Abajo , Calor , Masculino , Ratones , Proteínas de Unión al ARN/genética , Espermatocitos/metabolismo , Espermatogénesis/fisiología , Regulación hacia ArribaRESUMEN
This study aimed to evaluate the efficiency of the International Classification of Functioning, Disability, and Health (ICF) in stroke rehabilitation assessment in China and to identify correlations between the ICF and several commonly used clinical assessment instruments for stroke.In total, 52 hospitals and 5 premier rehabilitation and neurology research centers participated in this cross-sectional multicenter clinical study. A total of 2822 stroke patients admitted to a neurology or rehabilitation department of a participating medical center between July 2012 and June 2014 were included. The ICF checklist contains 4 parts with 128 two-level items: body functions, body structures, activities and participation, and environmental factors. We analyzed the results of ICF assessments and determined whether correlations existed between the various items of the ICF and several commonly used clinical assessment instruments.In all but 3 instances, the scores for the ICF-b-body function, ICF-s-body structure-degree of impairment, ICF-s-body structure-impairment location, ICF-d-activity performance, ICF-d-ability performance, ICF-e-facilitator, and ICF-e-barrier correlated significantly (Pâ<â.05) with the scores for the commonly used clinical assessment instruments.The ICF checklist is a new rehabilitation assessment instrument that is compatible with commonly used clinical assessment scales for stroke and can be used in combination with these scales.
Asunto(s)
Evaluación de la Discapacidad , Clasificación Internacional del Funcionamiento, de la Discapacidad y de la Salud , Evaluación de Resultado en la Atención de Salud/métodos , Rehabilitación de Accidente Cerebrovascular/estadística & datos numéricos , Accidente Cerebrovascular/fisiopatología , Actividades Cotidianas , Anciano , Lista de Verificación , Estudios Transversales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
OBJECTIVE(S): This study aimed to determine whether intrauterine placement of a novel composite material [copper (Cu) microparticles, low-density polyethylene, and methyl vinyl silicone rubber (Cu/LDPE/MVQ)] could prevent pregnancy in rabbits, and to evaluate the effects of Cu/LDPE/MVQ on the endometrial environment. STUDY DESIGN: Eighty sexually mature female rabbits were randomly divided into four groups (n=20 each group): control (sham-operated), LDPE/MVQ, Cu/LDPE/MVQ microcomposite, and bare Cu. Ten rabbits from each implant-bearing group were randomly selected for a mating experiment beginning 30 days after insertion. Pregnancy outcomes were observed 15 days after mating. Factors associated with endometrial bleeding and inflammation in the remaining rabbits in each group, and the surface conditions of the implants, were investigated 90 days post-insertion. RESULTS: The Cu (0 embryo) and Cu/LDPE/MVQ (0 embryo) groups had significantly fewer embryos than the LDPE/MVQ (1.0±0.6 embryos, p<.05) and sham-operated groups (4.1±1.3 embryos, p<.05). Compared with bare Cu, the Cu/LDPE/MVQ composite material was associated with considerable reductions in injuries and factors associated with abnormal endometrial bleeding and inflammation, such as matrix metalloproteinase 9 (MMP9) and prostaglandin E2 (PGE2). Additionally, the surface of implanted Cu/LDPE/MVQ remained much smoother than that of implanted bare Cu. CONCLUSION(S): This novel Cu-containing intrauterine device material exhibits a similar effect in prevention of pregnancy with bare copper, and lower levels of inflammatory markers. IMPLICATIONS: This study demonstrates the potential of the novel Cu/LDPE/MVQ microcomposite material as a future substitute for conventional intrauterine device materials.