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This study aimed to explore the influence of circFOXM1/miR-218-5p molecular axis in the proliferation, apoptosis and migration of glioma cells. The levels of circFOXM1 and miR-218-5p in glioma and adjacent tissues were tested by qRT-PCR. Cultured human glioma U251 cells were randomly split into groups: si-NC, si-circFOXM1, miR-NC, miR-218-5p, si-circFOXM1+anti-miR-NC, si-circFOXM1+anti-miR-218-5p. MTT method, plate clone formation, flow cytometry and Transwell experiments were utilized for detecting the proliferation, clone formation, apoptosis and migration of glioma cells. Dual-luciferase reporter experiment authenticated the targeted relation of circFOXM1 and miR-218-5p. Western blot tested the levels of E-cadherin and N-cadherin. CircFOXM1 was upregulated while miR-218-5p was low expressed in glioma tissues versus normal tissues. After circFOXM1 silence or miR-218-5p overexpression, miR-218-5p level was increased, and cell apoptosis rate and E-cadherin expression were enhancive, whereas cell proliferation, cell clone formation and migration abilities, and N-cadherin level were reduced. CircFOXM1 could affect miR-218-5 level by negative regulation. Furthermore, miR-218-5p silence could reverse the stimulative influence of si-circFOXM1 on apoptosis rate, and E-cadherin level, and the repressive effect on cell viability, cell number of colony formation and migration, and N-cadherin expression. Inhibition of circFOXM1 expression could block the proliferation, clone formation, and migration and induce apoptosis of glioma cells by upregulating miR-218-5p.
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Apoptosis , Movimiento Celular , Proteína Forkhead Box M1 , Regulación Neoplásica de la Expresión Génica , Glioma , MicroARNs , ARN Circular , Humanos , Apoptosis/genética , Neoplasias Encefálicas/patología , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Cadherinas/metabolismo , Cadherinas/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Proteína Forkhead Box M1/genética , Glioma/patología , Glioma/genética , Glioma/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , ARN Circular/genética , ARN Circular/metabolismoRESUMEN
BACKGROUND: DNA damage and DNA damage repair (DDR) are important therapeutic targets for triple-negative breast cancer (TNBC), a subtype with limited chemotherapy efficiency and poor outcome. However, the role of microRNAs in the therapy is emerging. In this study, we explored whether miR-26a-5p could act as BRCAness and enhance chemotherapy sensitivity in TNBC. METHODS: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-26a-5p in breast cancer tissues and cell lines. CCK-8 was used to measure drug sensitivity in concentration gradient and time gradient. Comet assay was used to detect DNA damage. Flow cytometry was performed to examine apoptosis. Moreover, we used western blot and immunofluorescence to detect biomarkers. Luciferase reporter assay was performed to verify the combination of miR-26a-5p and 3'UTR of target gene. Hormone deprivation and stimulation assay were used to validate the effect of hormone receptors on the expression of miR-26a-5p. Chromatin immunoprecipitation (ChIP) assays were used to verify the binding sites of ER-a or PR with the promoter of miR-26a-5p. Animal experiments were performed to the effect of miR-26a-5p on Cisplatin treatment. RESULTS: The expression of miR-26a-5p was significantly downregulated in TNBC. Overexpressing miR-26a-5p enhanced the Cisplatin-induced DNA damage and following apoptosis. Interestingly, miR-26a-5p promoted the expression of Fas without Cisplatin stimulating. It suggested that miR-26a-5p provided a hypersensitivity state of death receptor apoptosis and promoted the Cisplatin sensitivity of TNBC cells in vitro and in vivo. Besides, miR-26a-5p negatively regulated the expression of BARD1 and NABP1 and resulted in homologous recombination repair defect (HRD). Notably, overexpressing miR-26a-5p not only facilitated the Olaparib sensitivity of TNBC cells but also the combination of Cisplatin and Olaparib. Furthermore, hormone receptors functioned as transcription factors in the expression of miR-26a-5p, which explained the reasons that miR-26a-5p expressed lowest in TNBC. CONCLUSIONS: Taken together, we reveal the important role of miR-26a-5p in Cisplatin sensitivity and highlight its new mechanism in DNA damage and synthetic lethal.
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MicroARNs , Neoplasias de la Mama Triple Negativas , Humanos , Animales , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/metabolismo , Cisplatino/farmacología , Cisplatino/uso terapéutico , Línea Celular Tumoral , Proliferación Celular/genética , MicroARNs/genética , Proteínas Portadoras , HormonasRESUMEN
OBJECTIVE: Bone marrow mesenchymal stem cell (BMSC)-derived extracellular vesicles (EVs) have been demonstrated as a potential therapeutic agent in acute kidney injury (AKI). However, little is known about the mechanisms of action of BMSC-derived EVs in AKI. Based on this, our research was designed to investigate the mechanism behind BMSC-derived EVs controlling inflammation and pyroptosis during AKI. METHODS: Peripheral blood from AKI patients was used for detection of microRNA (miR)-223-3p, HDAC2, and SNRK expression. An AKI rat model was established, and HK-2 cell injury was induced by lipopolysaccharide (LPS) to establish a cellular model. Co-culture with BMSC-derived EVs and/or gain- and loss-of-function assays were conducted in LPS-treated HK-2 to evaluate the functions of BMSCs-EVs, miR-223-3p, HDAC2, and SNRK. AKI rats were simultaneously injected with EVs and short hairpin RNAs targeting SNRK. The interactions among miR-223-3p, HDAC2, and SNRK were evaluated by RIP, ChIP, and dual-luciferase gene reporter assays. RESULTS: Patients with AKI had low miR-223-3p and SNRK expression and high HDAC2 expression in peripheral blood. Mechanistically, miR-223-3p targeted HDAC2 to accelerate SNRK transcription. In LPS-treated HK-2 cells, BMSCs-EVs overexpressing miR-223-3p increased cell viability and diminished cell apoptosis, KIM-1, LDH, IL-1ß, IL-6, TNF-α, NLRP3, ASC, cleaved caspase-1, and IL-18 expression, and GSDMD cleavage, which was nullified by HDAC2 overexpression or SNRK silencing. In AKI rats, BMSCs-EV-shuttled miR-223-3p reduced CRE and BUN levels, apoptosis, inflammation, and pyroptosis, which was abrogated by SNRK silencing. CONCLUSION: Conclusively, BMSC-derived EV-encapsulated miR-223-3p mitigated AKI-induced inflammation and pyroptosis by targeting HDAC2 and promoting SNRK transcription.
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Lesión Renal Aguda , Vesículas Extracelulares , MicroARNs , Humanos , Animales , Ratas , Piroptosis , Lipopolisacáridos , Lesión Renal Aguda/terapia , Inflamación , MicroARNs/genética , Histona Desacetilasa 2/genéticaRESUMEN
OBJECTIVES: Observational studies have revealed that socioeconomic status is associated with cardiovascular health. However, the potential causal effect remains unclear. Hence, we aimed to investigate the causal relationship between household income status and genetic susceptibility to cardiovascular-related diseases using a bidirectional Mendelian randomization (MR) study. METHODS: An MR study based on a large-sample cohort of the European population from a publicly available genome-wide association study datasets was conducted using a random-effects inverse-variance weighting model as the main standard. Simultaneously, MR-Egger regression, weighted median, and maximum likelihood estimation were used as supplements. Sensitivity analysis, consisting of a heterogeneity test and horizontal pleiotropy test, was performed using Cochran's Q, MR-Egger intercept, and MR-PRESSO tests to ensure the reliability of the conclusion. RESULTS: The results suggested that higher household income tended to lower the risk of genetic susceptibility to myocardial infarction (OR: 0.503, 95% CI = 0.405-0.625, P < 0.001), hypertension (OR: 0.667, 95% CI = 0.522-0.851, P = 0.001), coronary artery disease (OR: 0.674, 95% CI = 0.509-0.893, P = 0.005), type 2 diabetes (OR: 0.642, 95% CI = 0.464-0.889, P = 0.007), heart failure (OR: 0.825, 95% CI = 0.709-0.960, P = 0.013), and ischemic stroke (OR: 0.801, 95% CI = 0.662-0.968, P = 0.022). In contrast, no association was evident with atrial fibrillation (OR: 0.970, 95% CI = 0.767-1.226, P = 0.798). The reverse MR study suggested a potentially negative trend between heart failure and household income status. A sensitivity analysis verified the reliability of the results. CONCLUSIONS: The results revealed that the population with higher household income tended to have a lower risk of genetic susceptibility to myocardial infarction and hypertension.
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Enfermedades Cardiovasculares , Diabetes Mellitus Tipo 2 , Insuficiencia Cardíaca , Hipertensión , Infarto del Miocardio , Humanos , Predisposición Genética a la Enfermedad , Estudio de Asociación del Genoma Completo , Análisis de la Aleatorización Mendeliana , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: Long noncoding RNAs (lncRNAs) are significant regulators for sepsis-associated acute kidney injury (AKI). Noncoding RNA activated by DNA damage (NORAD) is highly expressed in the serum of patients with neonatal sepsis. We aimed to reveal the role of NORAD in sepsis-associated AKI. METHODS AND RESULTS: In this study, we established an AKI mouse model by cecal ligation and puncture (CLP) method and used the lipopolysaccharide (LPS)-stimulated HK-2 cells as the in vitro model of AKI. We identified the upregulation of NORAD expression in AKI mice and LPS-treated HK-2 cells. Silencing of NORAD alleviated renal injury by suppressing inflammation and apoptosis in vivo. The influences of NORAD suppression on cell apoptosis and inflammatory response in LPS-treated HK-2 cells were investigated by TUNEL and western blotting. NORAD deficiency inhibited HK-2 cell apoptosis and relieved the inflammation. Moreover, we explored the underlying mechanism by which NORAD regulates HK-2 cells. MiR-577 was verified to directly bind to NORAD, and GOLPH3 was identified as a target downstream miR-577. In addition, GOLPH3 overexpression countervailed the impacts of NORAD downregulation on apoptosis and inflammation in vitro. CONCLUSIONS: Our findings revealed that NORAD knockdown alleviates kidney injury in mice and decreases the inflammatory response and apoptosis of LPS-stimulated HK-2 cells via the miR-577/GOLPH3 axis.
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Lesión Renal Aguda , MicroARNs , ARN Largo no Codificante , Sepsis , Lesión Renal Aguda/genética , Lesión Renal Aguda/metabolismo , Animales , Apoptosis/genética , Humanos , Riñón/metabolismo , Lipopolisacáridos/farmacología , Proteínas de la Membrana/genética , Ratones , MicroARNs/genética , MicroARNs/metabolismo , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Sepsis/genéticaRESUMEN
BACKGROUND: Vaccination is important in influenza prevention but the immune response wanes with age. The circadian nature of the immune system suggests that adjusting the time of vaccination may provide an opportunity to improve immunogenicity. Our previous cluster trial in Birmingham suggested differences between morning and afternoon vaccination for some strains in the influenza vaccine in older adults. Whether this effect is also seen in a younger age group with less likelihood of compromised immunity is unknown. We therefore conducted an individual-based randomized controlled trial in Guangzhou to test the hypothesis that influenza vaccination in the morning induces a stronger immune response in older adults than afternoon vaccination. We included adults in middle age to determine if the effect was also seen in younger age groups. RESULTS: Of the 418 participants randomised, 389 (93.1%, 191 middle-aged adults aged 50-60 years and 198 older adults aged 65-75 years) were followed up. Overall, there was no significant difference between the antibody titers (geometric mean /95% CI) after morning vs afternoon vaccination (A/H1N1: 39.9 (32.4, 49.1) vs. 33.0 (26.7, 40.7), p = 0.178; A/H3N2: 92.2 (82.8, 102.7) vs. 82.0 (73.8, 91.2), p = 0.091; B: 15.8 (13.9, 17.9) vs. 14.4 (12.8, 16.3), p = 0.092), respectively. However, in pre-specified subgroup analyses, post-vaccination titers for morning versus afternoon vaccination in the 65-75 years subgroup were (A/H1N1): 49.5 (36.7, 66.6) vs. 32.9 (24.7, 43.9), p = 0.050; (A/H3N2): 93.5 (80.6, 108.5) vs. 73.1 (62.9, 84.9), p = 0.021; (B): 16.6 (13.8, 20.1) vs. 14.4 (12.3, 17.0), p = 0.095, respectively. Among females, antibody titers for morning versus afternoon vaccination were (A/H1N1): 46.9 (35.6, 61.8) vs. 31.1 (23.8, 40.7), p = 0.030; (A/H3N2): 96.0 (83.5, 110.3) vs. 84.7 (74.4, 96.5), p = 0.176; (B): 14.8 (12.7, 17.3) vs. 13.0 (11.3, 14.9), p = 0.061, respectively. In the 50-60 years old subgroup and males, there were no significant differences between morning and afternoon vaccination. CONCLUSIONS: Morning vaccination may enhance the immunogenicity to influenza vaccine in adults aged over 65 and women. An intervention to modify vaccination programs to vaccinate older individuals in the morning is simple, cost free and feasible in most health systems.
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Cytoplasmic male sterility (CMS) is a common biological phenomenon used in hybrid production of peppers (Capsicum annuum L.). Although several restorer-of-fertility (Rf) genes of pepper CMS lines have been mapped, there is no report that the Rf gene with clear gene function has been isolated. Here, pepper CMS line HZ1A and its restorer line HZ1C were used to construct (HZ1A × HZ1C) F2 populations and map the Rf gene. A single dominant gene CaRfHZ conferred male fertility according to inheritance analysis. Using sterile plants from (HZ1A × HZ1C) F2 populations and bulked segregant analysis (BSA), the CaRfHZ gene was mapped between P06gInDel-66 and P06gInDel-89 on chromosome 6. This region spans 533.81 kb, where four genes are annotated according to Zunla-1 V2.0 gene models. Based on the analysis of genomic DNA sequences, gene expressions, and protein structures, Capana06g002968 was proposed as the strongest candidate for the CaRfHZ gene. Our results may help with hybrid pepper breeding and to elucidate the mechanism of male fertility restoration in peppers.
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Capsicum , Piper nigrum , Capsicum/genética , Fertilidad/genética , Genes de Plantas , Marcadores Genéticos , Piper nigrum/genética , Fitomejoramiento , Infertilidad Vegetal/genéticaRESUMEN
BACKGROUND: Since the outbreak of coronavirus disease 2019 in December 2019, more than 8 million cases have occurred worldwide as of June 16, 2020. However, it is important to distinguish COVID-19 from other respiratory infectious diseases, such as influenza. Here, we comparatively described the clinical characteristics of children with COVID-19 and paediatric patients with influenza. METHODS: In this retrospective, single-centre study, we reviewed the electronic medical records of 585 paediatric patients with COVID-19 or influenza in Wuhan Children's Hospital, China. Clinical and epidemiological characteristics, laboratory findings, and clinical outcomes were comparatively analysed. RESULTS: The median ages were 6.96 years (IQR, 2-10.81) for children with confirmed COVID-19, 2.67 years (IQR, 1.03-15.25) for those with influenza A and 3.67 years (IQR, 1.62-5.54) for those with influenza B. Fever was a symptom in 84 (34.7%) COVID-19 cases, 132 (70.21%) influenza A cases and 111 (74.50%) influenza B cases. The median length of stay (LOS) was 11 (8-15) days for paediatric COVID-19 patients, 4 (3-6) days for influenza A patients and 5 (3-6) days for influenza B patients. Twenty-six (13.98%) influenza A patients and 18 (12.59%) influenza B patients presented with decreased white blood cell counts, while 13 (5.33%) COVID-19 patients presented with decreased white blood cell counts. Eight (3.28%) COVID-19 patients, 23 (12.71%) influenza A patients and 21 (14.79%) influenza B patients experienced lymphocytopenia. Acute cardiac injury occurred in 18 (7.29%) COVID-19 patients, while 37 (19.68%) influenza A and 27 (18.12%) influenza B patients had acute cardiac injury. CONCLUSION: In this study, the illnesses of children with COVID-19 were demonstrated to be less severe than those of paediatric patients with influenza, and COVID-19 patients had milder illness and fewer complications.
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Tratamiento Farmacológico de COVID-19 , COVID-19/etiología , Gripe Humana/tratamiento farmacológico , Gripe Humana/etiología , Adolescente , COVID-19/epidemiología , Niño , Niño Hospitalizado , Preescolar , China/epidemiología , Comorbilidad , Femenino , Fiebre/epidemiología , Hospitales Pediátricos , Humanos , Lactante , Gripe Humana/epidemiología , Tiempo de Internación , Linfopenia/epidemiología , Linfopenia/virología , Masculino , Síndrome de Dificultad Respiratoria/tratamiento farmacológico , Síndrome de Dificultad Respiratoria/epidemiología , Síndrome de Dificultad Respiratoria/etiología , Síndrome de Dificultad Respiratoria/virología , Estudios RetrospectivosRESUMEN
BACKGROUND: Epidemiological studies have suggested that adipsin and visfatin are associated with the development of type 2 diabetes. This is the first study to investigate the effects of supplementation with purified anthocyanins on serum adipsin and visfatin in patients with prediabetes or newly diagnosed diabetes. METHODS: A total of 160 participants with prediabetes or newly diagnosed diabetes (40-75 years old) were given 320 mg anthocyanins or placebo daily for 12 weeks in a randomized trial. Serum adipsin, serum visfatin, lipids and glycated hemoglobin A1c (HbA1c) were measured. The areas under the curve (AUCs) for glucose, insulin and C-peptide were determined before-and after-treatment by a standard 3-h 75 g oral glucose tolerance test (OGTT). RESULTS: Relatively significant increases in serum adipsin (net change 0.15 µg/mL [0.03, 0.27], p = 0.018) and decreases in visfatin (-3.5 ng/mL [-6.69, -0.31], p = 0.032) were observed between the anthocyanins and placebo groups. We also observed significant improvements in HbA1c (-0.11% [-0.22, -0.11], p = 0.033), apolipoprotein A-1 (apo A-1) (0.12 g/L [0.03, 0.21], p = 0.012) and apolipoprotein B (apo B) (-0.07 g/L [-0.14, -0.01], p = 0.033) in response to the anthocyanins intervention. CONCLUSION: Purified anthocyanins supplementation for 12 weeks increased serum adipsin and decreased serum visfatin in patients with prediabetes or newly diagnosed diabetes. Trial registration ClinicalTrials.gov, identifier: NCT02689765.
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Diabetes Mellitus Tipo 2 , Estado Prediabético , Adulto , Anciano , Antocianinas , Glucemia , Factor D del Complemento , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hemoglobina Glucada , Humanos , Persona de Mediana Edad , Nicotinamida Fosforribosiltransferasa , Estado Prediabético/diagnóstico , Estado Prediabético/tratamiento farmacológicoRESUMEN
The number of coronavirus disease 2019 (COVID-19) cases has exceeded 10 million. However, little is known about the epidemiology and clinical characteristics of COVID-19 infants. We collected medical information of 46 confirmed patients (<1 year old) and retrospectively analyzed epidemiological history, clinical symptoms, and laboratory test results. The median age was 5 (interquartile range, 2-7) months. Sixteen cases had fever and 27 cases had cough. Moderate disease was present in 40 cases and cardiac injury occurred in 38 cases, following by liver dysfunction in 20 cases and lymphocytosis in no cases. Of all infant patients, 2 received invasive mechanical ventilation and 1 died with multiple organ dysfunction syndrome.
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Infecciones por Coronavirus/epidemiología , Neumonía Viral/epidemiología , Factores de Edad , Betacoronavirus/aislamiento & purificación , COVID-19 , China/epidemiología , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/terapia , Tos/terapia , Tos/virología , Femenino , Fiebre/terapia , Fiebre/virología , Humanos , Lactante , Masculino , Insuficiencia Multiorgánica/terapia , Insuficiencia Multiorgánica/virología , Pandemias , Neumonía Viral/diagnóstico , Neumonía Viral/terapia , Estudios Retrospectivos , SARS-CoV-2 , Resultado del TratamientoRESUMEN
BACKGROUND: Since December 2019, SARS-CoV-2 has extended to most parts of China with >80â 000 cases and to at least 100 countries with >60â 000 international cases as of 15 March 2020. Here we used a household cohort study to determine the features of household transmission of COVID-19. METHODS: A total of 105 index patients and 392 household contacts were enrolled. Both index patients and household members were tested by SARS-CoV-2 RT-PCR. Information on all recruited individuals was extracted from medical records and confirmed or supplemented by telephone interviews. The baseline characteristics of index cases and contact patients were described. Secondary attack rates of SARS-CoV-2 to contact members were computed and the risk factors for transmission within the household were estimated. RESULTS: Secondary transmission of SARS-CoV-2 developed in 64 of 392 household contacts (16.3%). The secondary attack rate to children was 4% compared with 17.1% for adults. The secondary attack rate to the contacts within the households with index patients quarantined by themselves since onset of symptoms was 0% compared with 16.9% for contacts without quarantined index patients. The secondary attack rate to contacts who were spouses of index cases was 27.8% compared with 17.3% for other adult members in the households. CONCLUSIONS: The secondary attack rate of SARS-CoV-2 in household is 16.3%. Age of household contacts and spousal relationship to the index case are risk factors for transmission of SARS-CoV-2 within a household. Quarantine of index patients at home since onset of symptoms is useful to prevent the transmission of SARS-Co-2 within a household.
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Infecciones por Coronavirus/transmisión , Composición Familiar , Neumonía Viral/transmisión , Adolescente , Adulto , Betacoronavirus , COVID-19 , Niño , Preescolar , Estudios de Cohortes , Infecciones Comunitarias Adquiridas/transmisión , Trazado de Contacto/estadística & datos numéricos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pandemias , SARS-CoV-2 , Factores de Tiempo , Adulto JovenRESUMEN
Although growing advances have been made in the regulation of lupus nephritis recently, lupus nephritis is still one of the major causes of death in SLE patients and the pathogenesis remains largely unknown. Therefore, exploring the pathological mechanisms is urgently needed for designing and developing novel therapeutic strategies for lupus nephritis. Human renal mesangial cells (HRMCs) were transfected with sh-NEAT1, miR-146b mimic, pcDNA-NEAT1, miR-146b inhibitor, or sh-TRAF6 to modify their expression. Lipopolysaccharide (LPS) was used to induce inflammatory injury. Cell viability was examined with CCK8. Apoptosis was determined by flow cytometry and Hoechst staining. qRT-PCR and western blot were used to analyze gene expression. The secretion of inflammatory cytokines was examined with ELISA. The bindings of NEAT1 with miR-146b and miR-146b with TRAF6 were tested by dual-luciferase reporter assay. NEAT1 was upregulated in LPS-treated HRMCs. Both the knockdown of NEAT1 and TRAF6 suppressed the LPS-induced inflammatory injury in HRMCs. NEAT1 directly targeted miR-146b to control miR-146b-mediated regulation of TRAF6 expression in HRMCs. NEAT1 promoted the expression of TRAF6 via targeting miR-146b to accelerate the LPS-mediated renal mesangial cell injury in HRMCs. Moreover, TRAF6 activated the NF-κB signaling in HRMCs. NEAT1 accelerated renal mesangial cell injury via directly targeting miR-146b, promoting the expression of TRAF6, and activating the NF-κB signaling in lupus nephritis. Our investigation elucidated novel pathological mechanisms and provided potential therapeutic targets for lupus nephritis.
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Péptidos y Proteínas de Señalización Intracelular/metabolismo , Riñón/metabolismo , Nefritis Lúpica/metabolismo , Células Mesangiales/metabolismo , MicroARNs/metabolismo , FN-kappa B/metabolismo , ARN Largo no Codificante/metabolismo , Adolescente , Adulto , Estudios de Casos y Controles , Humanos , Riñón/patología , Nefritis Lúpica/patología , Persona de Mediana Edad , ARN Largo no Codificante/genética , Transducción de Señal , Transfección , Adulto JovenRESUMEN
Growth differentiation factor 11 (GDF11), a key member of the TGF-ß superfamily, plays critical roles in various medical conditions. Recently, GDF11 was found to suppress the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway and protect against inflammation. This study aimed to investigate the role of GDF11 in the development of rheumatoid arthritis (RA). We demonstrated that GDF11 treatment antagonized TNF-α-induced inflammation in macrophages. Moreover, GDF11 inhibited the development of arthritis in the collagen-induced arthritis and collagen antibody-induced arthritis models. Local gene transfer of GDF11 via adeno-associated virus exerted therapeutic effects, while local knockdown of GDF11 exaggerated inflammation in our collagen-induced arthritis model, as detected by expression levels of inflammatory biomarkers and the destruction of joint structures. Additionally, the results from both in vitro experiments and luciferase reporter gene mouse experiments implied that the NF-κB pathway might play a critical role in the therapeutic effect of GDF11 in RA. This study presents GDF11 as a potential target for the treatment of inflammatory arthritis, including RA.-Li, W., Wang, W., Liu, L., Qu, R., Chen, X., Qiu, C., Li, J., Hayball, J., Liu, L., Chen, J., Wang, X., Pan, X., Zhao, Y. GDF11 antagonizes TNF-α-induced inflammation and protects against the development of inflammatory arthritis in mice.
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Artritis Experimental/prevención & control , Proteínas Morfogenéticas Óseas/metabolismo , Factores de Diferenciación de Crecimiento/metabolismo , Animales , Artritis Experimental/metabolismo , Artritis Experimental/patología , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Artritis Reumatoide/terapia , Proteínas Morfogenéticas Óseas/antagonistas & inhibidores , Proteínas Morfogenéticas Óseas/genética , Colágeno/inmunología , Técnicas de Silenciamiento del Gen , Técnicas de Transferencia de Gen , Terapia Genética , Factores de Diferenciación de Crecimiento/antagonistas & inhibidores , Factores de Diferenciación de Crecimiento/genética , Humanos , Mediadores de Inflamación/metabolismo , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , Ratones Transgénicos , FN-kappa B/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Transducción de Señal , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Osteoarthritis (OA) is a common chronic degenerative disease characterized by degeneration in the joints and subsequent destruction of cartilage and bone, yet much remains to be elucidated regarding its molecular mechanism. Ghrelin is a recently discovered neuropeptide with anti-inflammatory actions, but it is unknown whether ghrelin is involved in OA. Human primary chondrocyte and cartilage samples were collected from patients with OA, and the expression pattern of ghrelin was assessed. Human chondrocyte and cartilage samples were stimulated with IL-1ß and TNF-α, and exogenous ghrelin-alleviated disorganization of catabolism and anabolism were mediated by IL-1ß and TNF-α. Destabilization of the medial meniscus and anterior cruciate ligament transection models were established in wild-type mice that were administered ghrelin or PBS. Severity of inflammation and degeneration in the joints were determined by measuring the levels of various inflammatory cytokines and degeneration-associated molecules. Ghrelin down-regulated the production of various inflammatory cytokines, inhibited apoptosis of chondrocytes, decreased the levels of metalloproteinases (including matrix metalloproteinase-13 and a disintegrin and metalloproteinase with thrombospondin motif-5), and maintained the expression of critical matrix components, such as aggrecan and collagen 2. Moreover, suppression of the Akt signaling pathway and activation of NF-κB signaling in chondrocytes during OA development was antagonized by ghrelin administration. This supports the assessment of ghrelin as a potential therapeutic approach to treat degenerative cartilage diseases, including OA.-Qu, R., Chen, X., Wang, W., Qiu, C., Ban, M., Guo, L., Vasilev, K., Chen, J., Li, W., Zhao, Y. Ghrelin protects against osteoarthritis through interplay with Akt and NF-κB signaling pathways.
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Condrocitos/metabolismo , Ghrelina/farmacología , FN-kappa B/metabolismo , Osteoartritis/prevención & control , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Condrocitos/patología , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Metaloproteinasa 13 de la Matriz/genética , Metaloproteinasa 13 de la Matriz/metabolismo , Ratones , FN-kappa B/genética , Osteoartritis/genética , Osteoartritis/metabolismo , Osteoartritis/patología , Proteínas Proto-Oncogénicas c-akt/genética , Transducción de Señal/genética , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The current study explored the efficacy of an intra-articular (IA) injection of allogeneic adipose tissue-derived stem cells (ADSCs) combined with xanthan gum (XG) in a rat osteoarthritis (OA) model. We confirmed that XG significantly inproved proliferation of ADSCs in a dose dependent manner in vitro. The rat OA model was induced by an anterior cruciate ligament transection (ACLT), and at 4 weeks after surgery, rats were divided into four groups: the XG-ADSCs group, the ADSCs group, the XG group and the phosphate-buffered saline (PBS) group. A single dose of 1 × 106 allogeneic ADSCs suspended in 1% XG, ADSCs suspended in PBS, 1% XG alone or PBS alone was injected into the OA joint of rats in the respective treatment groups. Rats were sacrificed at 8 weeks after surgery. Treatment outcomes were evaluated by weight-bearing control of the hind limbs, gross morphological analysis, histological analysis and specific staining of articular cartilage, and measurement of inflammatory factors in synovial fluid. For the rats in the XG-ADSC-s and ADSCs-treated groups, the weight-bearing percentage of the right hind limb was significantly increased compared to that in the PBS group and was sustained over 4 weeks. However, the positive effect in the XG-ADSCs group was significantly greater than that in the ADSCs group. For the rats in the XG group, the efficacy decreased during the third week after surgery. The articular cartilage was relatively normal in the XG-ADSCs group, and moderate degeneration was observed in the ADSCs and XG groups. ADSCs and XG-ADSC treatments significantly decreased the concentrations of IL-1ß, TNF-α, MMP-3 and MMP-13 in synovial fluid; however, the attenuating effect of the XG-ADSCs treatment was significantly enhanced compared with that of the ADSCs treatment alone. These results indicate that a single IA injection of allogeneic ADSCs combined with XG efficiently attenuated OA progression with a therapeutic effect that was significantly greater than that of either ADSCs or XG alone. IA injection of XG-ADSCs might be an effective treatment for OA in humans.
Asunto(s)
Adipocitos/efectos de los fármacos , Antiinflamatorios no Esteroideos/farmacología , Osteoartritis/terapia , Polisacáridos Bacterianos/farmacología , Trasplante de Células Madre , Células Madre/efectos de los fármacos , Adipocitos/citología , Adipocitos/fisiología , Tejido Adiposo/citología , Tejido Adiposo/fisiología , Animales , Ligamento Cruzado Anterior/cirugía , Lesiones del Ligamento Cruzado Anterior/patología , Lesiones del Ligamento Cruzado Anterior/fisiopatología , Cartílago Articular/citología , Cartílago Articular/efectos de los fármacos , Diferenciación Celular , Citocinas/antagonistas & inhibidores , Citocinas/biosíntesis , Modelos Animales de Enfermedad , Miembro Posterior , Humanos , Inyecciones Intraarticulares , Masculino , Osteoartritis/patología , Osteoartritis/fisiopatología , Cultivo Primario de Células , Ratas , Ratas Wistar , Células Madre/citología , Células Madre/fisiología , Líquido Sinovial/química , Trasplante Homólogo , Soporte de PesoRESUMEN
Development of novel materials for enrichment of glycopeptides is the key to a comprehensive analysis of the glycoproteome, which is closely related to several major diseases and biomarker findings. We synthesized phenylboronic acid (PBA) bound to SiO2 microspheres by a thiol-ene click chemistry method (this material was denoted as click PBA) and used it to separate cis-diol-containing molecules and enrich glycopeptides in hydrophilic interaction chromatography mode. Successful preparation of click PBA was confirmed by elemental analysis, X-ray photoelectron spectroscopy, N2 adsorption-desorption isotherms, and high-resolution scanning electron microscopy. Click PBA showed stronger retention towards glycopeptides under alkaline, higher content of organic solution conditions than under acidic, higher content of organic solution or alkaline aqueous solution conditions. Click PBA exhibited high selectivity for both neutral and acidic glycopeptides, which could resist interference from 100 molar fold of bovine serum albumin digests. The high enrichment efficiency can be ascribed to the synergetic effects of affinity interaction and hydrophilic interaction. The application of click PBA to 1 µL human serum resulted in the identification of 101 unique glycosylation sites from 71 glycoproteins. Such material will facilitate comprehensive glycoproteome analysis.
Asunto(s)
Ácidos Borónicos/química , Glicopéptidos/química , Glicopéptidos/aislamiento & purificación , Microesferas , Animales , Ácidos Borónicos/síntesis química , Química Clic , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Espectrometría de Masas , Microscopía Electrónica de Rastreo , Albúmina Sérica BovinaRESUMEN
AIM: Microvesicles (MVs) are nanoscale membrane fragments released from virtually all cell types upon activation or apoptosis, and may contribute to the beneficial effects of stem cell therapy. In this study, we investigated the therapeutic effects of mesenchymal stem cell (MSC) derived MVs (MSC-MVs) on pulmonary artery hypertension (PAH) in rats. METHODS: MSC-MVs were isolated from rat bone marrow MSCs that were cultured in a serum-free conditioned medium. Transmission electron microscopy (TEM), flow cytometry and nanoparticle tracking analysis (NTA) were used to characterize the MVs. Adult SD rats were injected with monocrotaline (50 mg/kg, sc) to induce PAH. Three weeks later, the rats were intravenously injected with MSCs, MSC-MVs or saline for 2 weeks. At the end of treatments, the hemodynamic parameters and pathological right ventricular and pulmonary arterial remodeling were analyzed in each group. RESULTS: The MSC-MVs showed general morphologic characteristics of MVs and expressed annexin V and CD29 markers under TEM, and their size ranged from 40 to 300 nm. Intravenous injection of MSC-MVs or MSCs significantly ameliorated the mean pulmonary artery pressure (mPAP) and mean right ventricle pressure (mRVP) in PAH rats. Furthermore, intravenous injection of MSC-MVs or MSCs significantly decreased the right ventricle (RV) hypertrophy and pulmonary arteriole area index (AI) and thickness index (TI) in PAH rats. CONCLUSION: Intravenous injection of MSC-MVs or MSCs produces similar beneficial effects for treating PAH, and our results provide a basis for cell-free approach in stem cell therapy.
Asunto(s)
Micropartículas Derivadas de Células/fisiología , Hipertensión Pulmonar/fisiopatología , Hipertensión Pulmonar/terapia , Células Madre Mesenquimatosas/fisiología , Arteria Pulmonar/fisiología , Animales , Células Cultivadas , Ventrículos Cardíacos/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To explore the effect of transfection of adenovirus carrying hepatocyte growth factor (HGF) on pulmonary arterial hypertension (PAH) and endothelial cell membrane microparticles (EMP) in a rat model, and the underlying mechanism. METHODS: Forty healthy male SD rats were randomly divided into four groups, the normal control group (NOR group), monocrotaline (MCT)-induced pulmonary hypertension group (PAH group), HGF treatment of PAH group (HGF group and THGF group) each with 10 rats. NOR group and the PAH Group: intratracheal instillation of 0.2 ml PBS solution; HGF group: intratracheal instillation of 0.2 ml HGF one times; THGF Group: intratracheal instillation of 0.2 ml HGF one times, and then 1 week repeat again. Different interventions after 2 weeks, the rats was measured mean pulmonary arterial pressure, right ventricular hypertrophy index calculation, HE staining index of pulmonary arterial wall thickness, area index, plasma levels of endothelial cell microparticles. RESULTS: HGF intratracheal instillation after 2 weeks, HGF and THGF groups of SD rats mPAP, RVHI, TI, AI decreased significantly compared with PAH group (P < 0.05). PAH group was increased in particulate levels at different time points significantly higher levels of horizontal (P < 0.05). The EMP levels in HGF group which at 7 and 14 days after dosing were significantly decreased compared with PAH.It still higher than the NOR group (P < 0.05). And after administration of 7 days, 14 days, the EMP level of HGF group was significantly lower than before administration (P < 0.05). CONCLUSIONS: Thought airway instillation transfected HGF, pulmonary artery pressure can reduce greater degree, but can't achieve complete reversal. It can inhibit the pulmonary artery wall thickening and maintain effective lumen area, delaying the right ventricular hypertrophy by reducing the membrane particles within the lung, thereby promoting endothelial cell repair to achieve the goal of intervention in pulmonary hypertension.
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Endotelio Vascular/efectos de los fármacos , Factor de Crecimiento de Hepatocito/genética , Hipertensión Pulmonar/tratamiento farmacológico , Arteria Pulmonar/efectos de los fármacos , Adenoviridae/genética , Animales , Presión Sanguínea/efectos de los fármacos , Modelos Animales de Enfermedad , Células Endoteliales/efectos de los fármacos , Factor de Crecimiento de Hepatocito/farmacología , Hipertensión Pulmonar/inducido químicamente , Hipertensión Pulmonar/fisiopatología , Masculino , Monocrotalina/efectos adversos , Arteria Pulmonar/patología , Ratas , Ratas Sprague-Dawley , TransfecciónRESUMEN
This study aims to investigate the positive effects of ultra-high pressure assisted acid extraction (UPAAE) on both physicochemical properties and antioxidant activities of hawthorn pectin. The basic indicators, structure characterization, and antioxidant activities were measured, which could indicate the disadvantages and advantages among traditional water extraction (WE), acid extraction (AE), and UPAAE. The results show that the hawthorn pectin of UPAAE has a decrease in esterification degree, protein content, and total polyphenols, but has an increase in total galacturonic acid aldehyde compared to the hawthorn pectin of AE. In the Fourier Transform Infrared Spectroscopy (FT-IR) and Scanning Electron Microscopy (SEM) analyses, the hawthorn of UPAAE has typical pectin absorption peaks in the FT-IR spectrum and a distinct layered structure in the SEM surface image. The ion chromatography profiles show that the molar ratio of galacturonic acid to arabinose in the hawthorn pectin of UPAAE increases and 5.50 µg/mg ribose appears compared to the pectin of AE and WE. The high performance gel permeation chromatography (HPGPC) profile indicates that the molecular weight distribution of hawthorn pectin of UPAAE is more concentrated and has the highest molecular weight compared to the pectin of the other two extraction methods. In the vitro antioxidant activity analysis, the pectin of UPAAE exhibits the highest scavenging rate against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals (93.70%), which is close to the scavenging rate of vitamin C (96.30%). These findings demonstrated that UPAAE is a more efficient and environmentally friendly method for pectin extraction from hawthorn. It is also an effective way to enhance its antioxidant activity, which has great application prospects in the food industries.