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Pediatric-onset colitis and inflammatory bowel disease (IBD) have significant effects on the growth of infants and children, but the etiopathogenesis underlying disease subtypes remains incompletely understood. Here, we report single-cell clustering, immune phenotyping, and risk gene analysis for children with undifferentiated colitis, Crohn's disease, and ulcerative colitis. We demonstrate disease-specific characteristics, as well as common pathogenesis marked by impaired cyclic AMP (cAMP)-response signaling. Specifically, infiltration of PDE4B- and TNF-expressing macrophages, decreased abundance of CD39-expressing intraepithelial T cells, and platelet aggregation and release of 5-hydroxytryptamine at the colonic mucosae were common in colitis and IBD patients. Targeting these pathways by using the phosphodiesterase inhibitor dipyridamole restored immune homeostasis and improved colitis symptoms in a pilot study. In summary, comprehensive analysis of the colonic mucosae has uncovered common pathogenesis and therapeutic targets for children with colitis and IBD.
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Enfermedades Inflamatorias del Intestino/patología , Enfermedades Inflamatorias del Intestino/terapia , Mucosa Intestinal/patología , Antígenos CD/metabolismo , Apirasa/metabolismo , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Muerte Celular/efectos de los fármacos , Microambiente Celular/efectos de los fármacos , Niño , Estudios de Cohortes , Colon/patología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/metabolismo , Dipiridamol/farmacología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Células Epiteliales/patología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Predisposición Genética a la Enfermedad , Homeostasis/efectos de los fármacos , Humanos , Inmunoglobulina G/sangre , Memoria Inmunológica , Inflamación/patología , Enfermedades Inflamatorias del Intestino/sangre , Enfermedades Inflamatorias del Intestino/genética , Interferón Tipo I/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Metilprednisolona/farmacología , Células Mieloides/efectos de los fármacos , Células Mieloides/metabolismoRESUMEN
Fluorescent RNAs, aptamers that bind and activate small fluorogenic dyes, have provided a particularly attractive approach to visualizing RNAs in live cells. However, the simultaneous imaging of multiple RNAs remains challenging due to a lack of bright and stable fluorescent RNAs with bio-orthogonality and suitable spectral properties. Here, we develop the Clivias, a series of small, monomeric and stable orange-to-red fluorescent RNAs with large Stokes shifts of up to 108 nm, enabling the simple and robust imaging of RNA with minimal perturbation of the target RNA's localization and functionality. In combination with Pepper fluorescent RNAs, the Clivias enable the single-excitation two-emission dual-color imaging of cellular RNAs and genomic loci. Clivias can also be used to detect RNA-protein interactions by bioluminescent imaging both in live cells and in vivo. We believe that these large Stokes shift fluorescent RNAs will be useful tools for the tracking and quantification of multiple RNAs in diverse biological processes.
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Aptámeros de Nucleótidos , Colorantes Fluorescentes , ARN , Microscopía Fluorescente , Aptámeros de Nucleótidos/genéticaRESUMEN
The discovery and identification of broad-spectrum antiviral drugs are of great significance for blocking the spread of pathogenic viruses and corresponding variants of concern. Herein, we proposed a plasmonic imaging-based strategy for assessing the efficacy of potential broad-spectrum antiviral drugs targeting the N-terminal domain of a nucleocapsid protein (NTD) and nucleic acid (NA) interactions. With NTD and NA conjugated gold nanoparticles as core and satellite nanoprobes, respectively, we found that the multivalent binding interactions could drive the formation of core-satellite nanostructures with enhanced scattering brightness due to the plasmonic coupling effect. The core-satellite assembly can be suppressed in the presence of antiviral drugs targeting the NTD-NA interactions, allowing the drug efficacy analysis by detecting the dose-dependent changes in the scattering brightness by plasmonic imaging. By quantifying the changes in the scattering brightness of plasmonic nanoprobes, we uncovered that the constructed multivalent weak interactions displayed a 500-fold enhancement in affinity as compared with the monovalent NTD-NA interactions. We demonstrated the plasmonic imaging-based strategy for evaluating the efficacy of a potential broad-spectrum drug, PJ34, that can target the NTD-NA interactions, with the IC50 as 24.35 and 14.64 µM for SARS-CoV-2 and SARS-CoV, respectively. Moreover, we discovered that ceftazidime holds the potential as a candidate drug to inhibit the NTD-NA interactions with an IC50 of 22.08 µM from molecular docking and plasmonic imaging-based drug analysis. Finally, we validated that the potential antiviral drug, 5-benzyloxygramine, which can induce the abnormal dimerization of nucleocapsid proteins, is effective for SARS-CoV-2, but not effective against SARS-CoV. All these demonstrations indicated that the plasmonic imaging-based strategy is robust and can be used as a powerful strategy for the discovery and identification of broad-spectrum drugs targeting the evolutionarily conserved viral proteins.
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Antivirales , Oro , Nanopartículas del Metal , SARS-CoV-2 , Antivirales/farmacología , Antivirales/química , Oro/química , Nanopartículas del Metal/química , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/química , Humanos , Proteínas de la Nucleocápside de Coronavirus/química , Proteínas de la Nucleocápside de Coronavirus/metabolismo , Ácidos Nucleicos/química , Ácidos Nucleicos/metabolismo , Tratamiento Farmacológico de COVID-19 , Dominios Proteicos , FosfoproteínasRESUMEN
BACKGROUND: To investigate the changes in the unhealthy eye-related behaviors of junior middle school students during the COVID-19 pandemic and the double reduction policy and its relationship with myopia. METHODS: Data were obtained from the 2019-2022 Tianjin Children and Youth Myopia, Common Diseases and Health Influencing Factors Survey. Latent profile analysis (LPA) and a generalized linear model (GLM) were applied to analyze the effect of eye-related behavior classes on myopia. RESULTS: A total of 2508 junior middle school students were included. The types of eye-related behavior were categorized into the medium-healthy behavior group, heavy academic burden and near-eye behavior group, insufficient lighting group and high-healthy behavior group. Students with heavy academic burdens and near-eye behavior were more likely to develop myopia than were those in the high-healthy group (OR = 1.466, 95% CI = 1.203-1.787; P < 0.001). CONCLUSIONS: The dual reduction policy has a positive effect on improving unhealthy eye-related behaviors, and the prevention and control of myopia through the use of different combinations of eye-related behaviors are heterogeneous among junior middle school students. In the post-COVID-19 period, we should continue to implement a double reduction policy and formulate targeted eye-related behavior strategies to provide an important reference for the prevention and control of myopia among children and adolescents during public health emergencies in the future.
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COVID-19 , Miopía , Estudiantes , Humanos , COVID-19/epidemiología , COVID-19/prevención & control , COVID-19/psicología , Miopía/epidemiología , Miopía/psicología , Miopía/prevención & control , Masculino , Femenino , Adolescente , Niño , Estudiantes/psicología , Estudiantes/estadística & datos numéricos , China/epidemiología , Conductas Relacionadas con la Salud , Pandemias , Instituciones Académicas , Encuestas y CuestionariosRESUMEN
Since 2009, China has made large investments in strengthening the primary healthcare system. This study aimed to examine the trends in the number and distribution of health resources in rural China following the health system reform and to decompose the sources of inequalities. Data were collected from standardized reports compiled by each county in rural China and compiled by the National Health Commission and Bureau of Statistics. From the findings of this empirical study, resource allocation per capita for primary health care (PHC) improved gradually from 2008 to 2014. The distribution of beds across counties (ranked by level of economic development) was relatively equitable. However, the concentration curve analysis indicated that the distribution of primary care professionals remained skewed in favour of wealthier and more urbanised counties. Economic status was proved to be a major contributor to the inequality of health human resource. China's primary care reforms resulted in simultaneously improved supply of PHC resources as well as pro-rich inequality in distribution of the workforce. To advance equality in health resource allocation, greater attention should be paid to the substantial inequality of economic status within counties.
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The wild strawberry (Fragaria vesca L.; F. vesca) represents a resilient and extensively studied model organism. While the AP2/ERF gene family plays a pivotal role in plant development, its exploration within F. vesca remains limited. In this study, we characterized the AP2/ERF gene family in wild strawberries using the recently released genomic data (F. vesca V6.0). We conducted an analysis of the gene family expansion pattern, we examined gene expression in stem segments and leaves under cold conditions, and we explored its functional attributes. Our investigation revealed that the FvAP2/ERF family comprises 86 genes distributed among four subfamilies: AP2 (17), RAV (6), ERF (62), and Soloist (1). Tandem and segmental duplications significantly contributed to the growth of this gene family. Furthermore, predictive analysis identified several cis-acting elements in the promoter region associated with meristematic tissue expression, hormone regulation, and resistance modulation. Transcriptomic analysis under cold stress unveiled diverse responses among multiple FvAP2/ERFs in stem segments and leaves. Real-time fluorescence quantitative reverse transcription PCR (RT-qPCR) results confirmed elevated expression levels of select genes following the cold treatment. Additionally, overexpression of FvERF23 in Arabidopsis enhanced cold tolerance, resulting in significantly increased fresh weight and root length compared to the wild-type control. These findings lay the foundation for further exploration into the functional roles of FvAP2/ERF genes.
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Fragaria , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Familia de Multigenes , Proteínas de Plantas , Fragaria/genética , Fragaria/metabolismo , Fragaria/crecimiento & desarrollo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Filogenia , Genoma de Planta , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Respuesta al Choque por Frío/genética , Regiones Promotoras GenéticasRESUMEN
The NAC (NAM, ATAF1/2, CUC2) family of transcription factors (TFs) is a vital transcription factor family of plants. It controls multiple parts of plant development, tissue formation, and abiotic stress response. We cloned the FvNAC29 gene from Fragaria vesca (a diploid strawberry) for this research. There is a conserved NAM structural domain in the FvNAC29 protein. The highest homology between FvNAC29 and PaNAC1 was found by phylogenetic tree analysis. Subcellular localization revealed that FvNAC29 is localized onto the nucleus. Compared to other tissues, the expression level of FvNAC29 was higher in young leaves and roots. In addition, Arabidopsis plants overexpressing FvNAC29 had higher cold and high-salinity tolerance than the wild type (WT) and unloaded line with empty vector (UL). The proline and chlorophyll contents of transgenic Arabidopsis plants, along with the activities of the antioxidant enzymes like catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) under 200 mM NaCl treatment or -8 °C treatment, were higher than those activities of the control. Meanwhile, malondialdehyde (MDA) and the reactive oxygen species (ROS) content were higher in the WT and UL lines. FvNAC29 improves transgenic plant resistance to cold and salt stress by regulating the expression levels of AtRD29a, AtCCA1, AtP5CS1, and AtSnRK2.4. It also improves the potential to tolerate cold stress by positively regulating the expression levels of AtCBF1, AtCBF4, AtCOR15a, and AtCOR47. These findings suggest that FvNAC29 may be related to the processes and the molecular mechanisms of F. vesca response to high-salinity stress and LT stress, providing a comprehensive understanding of the NAC TFs.
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Arabidopsis , Fragaria , Arabidopsis/genética , Fragaria/genética , Filogenia , Peroxidasas , AntioxidantesRESUMEN
Pepper fluorescent RNAs are a recently reported bright, stable and multicolor fluorogenic aptamer tag that enable imaging of diverse RNAs in live cells. To investigate the molecular basis of the superior properties of Pepper, we determined the structures of complexes of Pepper aptamer bound with its cognate HBC or HBC-like fluorophores at high resolution by X-ray crystallography. The Pepper aptamer folds in a monomeric non-G-quadruplex tuning-fork-like architecture composed of a helix and one protruded junction region. The near-planar fluorophore molecule intercalates in the middle of the structure and is sandwiched between one non-G-quadruplex base quadruple and one noncanonical G·U wobble helical base pair. In addition, structure-based mutational analysis is evaluated by in vitro and live-cell fluorogenic detection. Taken together, our research provides a structural basis for demystifying the fluorescence activation mechanism of Pepper aptamer and for further improvement of its future application in RNA visualization.
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Aptámeros de Nucleótidos/química , Colorantes Fluorescentes/química , ARN/química , Emparejamiento Base , Secuencia de Bases , Sitios de Unión , Cristalografía por Rayos X , G-Cuádruplex , Células HEK293 , Humanos , Técnicas In Vitro , Estructura Molecular , Mutación , Relación Estructura-ActividadRESUMEN
Au-Fe3O4 multifunctional nanoparticles (NPs) were synthesized and integrated with lateral flow immunoassay (LFIA) for dual-mode detection of Salmonella typhimurium. The Au-Fe3O4 NPs not only combined excellent local surface plasmon resonance characteristics and superparamagnetic properties, but also exhibited good photothermal effect. In the detection, antibody-conjugated Au-Fe3O4 NPs first captured S. typhimurium from complex matrix, which was then loaded on the LFIA strip and trapped by the T-line. By observing the color bands with the naked eyes, qualitative detection was performed free of instrument. By measuring the photothermal signal, quantification was achieved with a portable infrared thermal camera. The introduction of magnetic separation achieved the enrichment and purification of target bacteria, thus enhancing the detection sensitivity and reducing interference. This dual-mode LFIA achieved a visual detection limit of 5 × 105 CFU/mL and a photothermal detection limit of 5 × 104 CFU/mL. Compared with traditional Au-based LFIA, this dual-mode LFIA increased the detection sensitivity by 2 orders of magnitude and could be directly applied to unprocessed milk sample. Besides, this dual-mode LFIA showed good reproducibility and specificity. The intra-assay and inter-assay variation coefficients were 3.0% and 7.9%, and with this dual-mode LFIA, other bacteria hardly produced distinguishable signals. Thus, the Au-Fe3O4 NPs-based LFIA has potential to increase the efficiency of pandemic prevention and control. Au-Fe3O4 nanoparticle proved to be a promising alternative reporter for LFIA, achieving multifunctions: target purification, target enrichment, visual qualitation, and instrumental quantification, which improved the limitations of traditional LFIA.
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Nanopartículas del Metal , Nanopartículas Multifuncionales , Salmonella typhimurium , Colorimetría , Reproducibilidad de los Resultados , InmunoensayoRESUMEN
The dynamics of soil organic carbon (SOC) play a critical role in modulating global warming. However, the long-term spatiotemporal changes of SOC at large scale, and the impacts of driving forces remain unclear. In this study, we investigated the dynamics of SOC in different soil layers across China through the1980s to 2010s using a machine learning approach and quantified the impacts of the key factors based on factorial simulation experiments.Our results showed that the latest (2000-2014) SOC stock in the first meter soil (SOC100 ) was 80.68 ± 3.49 Pg C, of which 42.6% was stored in the top 20 cm, sequestrating carbon with a rate of 30.80 ± 12.37 g C m-2 yr-1 since the 1980s. Our experiments focusing on the recent two periods (2000s and 2010s) revealed that climate change exerted the largest relative contributions to SOC dynamics in both layers and warming or drying can result in SOC loss. However, the influence of climate change weakened with soil depth, while the opposite for vegetation growth. Relationships between SOC and forest canopy height further confirmed this strengthened impact of vegetation with soil depth and highlighted the carbon sink function of deep soil in mature forest. Moreover, our estimates suggested that SOC dynamics in 71% of topsoil were controlled by climate change and its coupled influence with environmental variation (CE). Meanwhile, CE and the combined influence of climate change and vegetation growth dominated the SOC dynamics in 82.05% of the first meter soil. Additionally, the national cropland topsoil organic carbon increased with a rate of 23.6 ± 7.6 g C m-2 yr-1 since the 1980s, and the widely applied nitrogenous fertilizer was a key stimulus. Overall, our study extended the knowledge about the dynamics of SOC and deepened our understanding about the impacts of the primary factors.
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Carbono , Suelo , Carbono/análisis , Secuestro de Carbono , China , Aprendizaje AutomáticoRESUMEN
AIMS AND OBJECTIVES: This study aims to explore the convenience and practicality of the method of establishing buttonholes with indwelling trocars. BACKGROUND: Compared with rope-ladder technique, buttonhole cannulation has better applicability for haemodialysis patients, and the method of buttonhole establishment is particularly important. DESIGN: Prospective, observational and cohort study. METHODS: We conducted a 12-month observational study on 30 patients who used sharp needles to establish buttonholes and 33 patients who used indwelling trocars to establish buttonholes, and compared the differences between the two groups in the buttonhole formation time, patient-reported cannulation pain, buttonhole success rate, AVF-related infection and patency of AVF. The study adhered to STROBE guidelines. RESULTS: The buttonhole formation time of the indwelling trocar group is less than that of the sharp needle group (5.48 ± 0.51 vs. 23.23 ± 3.07, p < .01). When the buttonhole was used for the first time, the cannulation pain of the indwelling trocar group was less than that of the sharp needle group (4.94 ± 1.50 vs. 6.03 ± 1.13, p = .002), but there was no significant difference in this result after 1 month (sharp needle group with 4.50 ± 0.94 vs. indwelling trocar group with 4.12 ± 1.19, p = .169). Compared with all the buttonholes in the indwelling trocar group were established successfully, there were 5 unsuccessful buttonholes in the sharp needle group (p = .02). Three cases of unsuccessful buttonhole patients in the sharp needle group developed AVF-related infection, this result did not appear in the indwelling trocar group (p = .102). The difference in arterial pressure and venous pressure between the two groups was not significant (p = .061, p = .222). CONCLUSIONS: Our new method can help buttonhole patients get less track formation time, less cannulation pain, and less fistula infection, thereby bringing them a more comfortable dialysis experience. RELEVANCE TO CLINICAL PRACTICE: The new method used in this study not only meets the needs of patients to protect AVF, but also provides convenience for clinical work. In addition, this study analyzed the causes of the AVF infection in buttonhole patients, and provided directions for future research.
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Derivación Arteriovenosa Quirúrgica , Derivación Arteriovenosa Quirúrgica/métodos , Estudios de Cohortes , Humanos , Dolor , Estudios Prospectivos , Diálisis Renal , Instrumentos QuirúrgicosRESUMEN
Crude glycerol, a by-product of biodiesel production, was employed as the carbon source to produce lipase using Pichia pastoris. Under identical fermentation conditions, cell growth and lipase activity were improved using crude glycerol instead of pure glycerol. The impacts of crude glycerol impurities (methyl ester, grease, glycerol, methanol, and metal ions Na+, Ca2+, and Fe3+) on lipase production were investigated. Impurities accelerated P. pastoris entering the stationary phase. Na+, Ca2+, and grease in waste crude glycerol were the main factors influencing higher lipase activity. Through response surface optimization of Ca2+, Na+, and grease concentrations, lipase activity reached 1437 U/mL (15,977 U/mg), which was 2.5 times that of the control. This study highlights the economical and highly efficient valorization of crude glycerol, demonstrating its possible utilization as a carbon source to produce lipase by P. pastoris without pretreatment.
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Medios de Cultivo/farmacología , Proteínas Fúngicas , Glicerol/farmacología , Lipasa , Rhizomucor/genética , Saccharomycetales/crecimiento & desarrollo , Medios de Cultivo/química , Proteínas Fúngicas/biosíntesis , Proteínas Fúngicas/genética , Glicerol/química , Lipasa/biosíntesis , Lipasa/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Rhizomucor/enzimología , Saccharomycetales/genéticaRESUMEN
OBJECTIVES: To study the predictive factors for glucocorticoid therapy by analyzing the association between the clinical features and treatment regimens in children with eosinophilic gastroenteritis. METHODS: A retrospective analysis was performed on the medical data of 182 children with eosinophilic gastroenteritis who were admitted to Guangzhou Women and Children's Medical Center from January 2012 to December 2020. According to whether glucocorticoids were used, these children were divided into a glucocorticoid treatment group and a control group. The two groups were compared in terms of age, history of allergy, clinical symptoms, laboratory examination results, endoscopic findings, and pathological results of gastrointestinal mucosa. A multivariate logistic regression analysis was performed for the results with statistical significance. RESULTS: Of the 182 children, 36 (19.8%) received glucocorticoid therapy. The rates of hematochezia, anemia, and mucosal ulceration/luminal stenosis under endoscopy and the mucosal eosinophil infiltration count were significantly higher in the glucocorticoid treatment group than those in the control group (P<0.05). The serum albumin level in the glucocorticoid treatment group was significantly lower than that in the control group (P<0.05). The multivariate logistic regression analysis showed that mucosal ulceration/luminal stenosis under endoscopy (OR=10.830, 95%CI: 3.090-37.961, P<0.001) and the increased mucosal eosinophil infiltration count (OR=0.967, 95%CI: 0.941-0.993, P=0.015) were predictive factors for glucocorticoid therapy in children with eosinophil gastroenteritis. CONCLUSIONS: Mucosal ulceration/luminal stenosis under endoscopy or a significant increase in the mucosal eosinophil infiltration count based on pathology suggests that glucocorticoid therapy can be considered in children with eosinophil gastroenteritis.
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Enteritis , Eosinofilia , Niño , Enteritis/tratamiento farmacológico , Eosinofilia/tratamiento farmacológico , Femenino , Gastritis , Glucocorticoides/uso terapéutico , Humanos , Estudios RetrospectivosRESUMEN
BACKGROUND Postextubation distress is detrimental to the prognosis of critically ill patients with successful spontaneous breathing trial. The known risk factors of failed weaning are associated with the heart, lungs, and diaphragm. The aim of this study was to explore the role of a combined model including indicators of heart, lung, and diaphragm ultrasound in predicting the weaning outcome. MATERIAL AND METHODS Patients' clinical data and ultrasonic features of heart, lungs, and diaphragm were recorded. Patients were included in either the failed weaning group (n=24) or the successful weaning group (n=81). The association of potential variables with the risk of weaning failure was determined using multivariate logistic regression analysis. The accuracy of potential indicators for predicting the weaning outcome were evaluated and a multiindicator combined model was established to improve the predictive accuracy. RESULTS Brain natriuretic peptide (odds ratio [OR]=1.120, P=0.004), left-atrial pressure (LAP) (OR=1.333, P=0.005), lung ultrasound score (LUS) (OR=1.736, P=0.001), and hemidiaphragm dysfunction (OR=3.942, P=0.014) were associated with an increased risk of weaning failure. However, all of these indicators could not accurately predict the weaning outcome independently (all areas under the curve [AUCs] <0.9). The combination of LAP, LUS, and hemidiaphragm dysfunction showed the highest AUC (AUC=0.919). CONCLUSIONS The combined model including LAP, LUS, and hemidiaphragm dysfunction were the most accurate method for the prediction.
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Diafragma/diagnóstico por imagen , Corazón/diagnóstico por imagen , Pulmón/diagnóstico por imagen , Respiración Artificial , Ultrasonografía/métodos , Desconexión del Ventilador , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Análisis MultivarianteRESUMEN
Long noncoding RNAs (lncRNAs) have been confirmed to be aberrantly expressed in various diseases including tumors. Recently, a new tumor-related lncRNA, lncRNA TRPM2 antisense RNA (TRPM2-AS), was shown to be involved in many tumors, such as lung cancer and breast cancer. However, the expression and role of TRPM2-AS in the development of gastric cancer (GC) have not been elucidated. In the current study, we provided evidence that the expression levels of TRPM2-AS were increased in both GC tissues and cell lines. We also showed that overexpression of TRPM2-AS was modulated by ELK1, a transcription factor. The results of clinical assays showed that higher expressions of TRPM2-AS were significantly related with invasion depth, TNM stage, lymphatic metastasis, and shorter overall survival. Further clinical assays using multivariate analysis suggested that TRPM2-AS expression was an independent prognostic factor in patients with GC. Functional experiments illustrated that depression of TRPM2-AS suppressed proliferation, migration, and invasion in GC cells. In terms of mechanism, we found that TRPM2-AS directly inhibited miR-195, which targeted the 3'-untranslated region of high-mobility group AT-hook 1 (HMGA1) messenger RNA. Overall, these findings revealed that ELK1-induced overexpression of TRPM2-AS promoted the development and progression of GC in part through miR-195/HMGA1 signaling axis, and established its candidacy as a new cancer biomarker for GC patients.
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Proteína HMGA1a/genética , MicroARNs/genética , ARN Largo no Codificante/biosíntesis , Neoplasias Gástricas/genética , Proteína Elk-1 con Dominio ets/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Proteína HMGA1a/biosíntesis , Humanos , Metástasis Linfática/genética , Masculino , MicroARNs/biosíntesis , Persona de Mediana Edad , Invasividad Neoplásica/genética , Pronóstico , Regiones Promotoras Genéticas/genética , Interferencia de ARN , ARN Largo no Codificante/genética , ARN Interferente Pequeño/genética , Transducción de Señal/genética , Neoplasias Gástricas/patologíaRESUMEN
Alzheimer's disease (AD) is the most prevalent form of dementia worldwide and is an emerging global epidemic. Active and passive immune therapies targeting beta amyloid (Aß) have shown very limited evidence in human studies of clinical benefits from these approaches. Epidemiological studies have shown that subjects with type 2 diabetes (T2D) are at higher risk of developing AD. However, whether and how these two conditions are causally linked is unknown. With the purpose of confirming the relationship between T2D and AD, this study specifically focused on effects of insulin in an in vitro model of the human blood-brain barrier (BBB) and on potential mechanisms of action in the treatment of AD. By using a series of assays to establish a BBB model, we demonstrated that insulin treatment alone could induce the increase of brain endothelial barrier properties. The transcriptional response of hCMEC/D3 cells to activation with different concentrations of insulin was determined by RT-PCR, and expression levels of genes involved in the control of barrier permeability, including inter-brain endothelial junctions, integrin-focal adhesions complexes, and transporter system, were found to be altered by the treatment. Notably, the influence of insulin on expression of the ATP-binding cassette (ABC) transporter which contributes to the clearance of Aß was investigated. Insulin up-regulated adherens junction and tight junction transmembrane proteins, as well as the ABC transporter. By treatment with insulin, the models have major advantages: it is fast, it has low cost, it is fit for considerable samples, and its conditions are under control.
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Transportadoras de Casetes de Unión a ATP/metabolismo , Barrera Hematoencefálica/metabolismo , Encéfalo/metabolismo , Permeabilidad de la Membrana Celular/efectos de los fármacos , Endotelio Vascular/metabolismo , Insulina/farmacología , Transcriptoma/efectos de los fármacos , Transportadoras de Casetes de Unión a ATP/genética , Péptidos beta-Amiloides/metabolismo , Transporte Biológico , Barrera Hematoencefálica/efectos de los fármacos , Encéfalo/citología , Encéfalo/efectos de los fármacos , Células Cultivadas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Regulación de la Expresión Génica , Humanos , Hipoglucemiantes/farmacología , Técnicas In Vitro , Modelos BiológicosRESUMEN
BACKGROUND AND PURPOSE: Inflammatory reaction plays a crucial role in cerebral ischemia reperfusion (IR) injury. It has been shown that activated microglia long-term existed in cerebral ischemia and induced second injury. Therefore, we hypothesize that prepared phosphatidylserine (PS)-modified microbubbles (PS-MBs) combined with ultrasound-targeted microbubble destruction (UTMD) can safely open the blood-brain barrier (BBB) and target activated microglia for inflammatory area in the later stage of ischemia reperfusion. METHODS: To verify our hypothesis, rat model of IR was established, then the change of activated microglia/macrophage (M/M) and permeability of BBB at 1, 7, 14, and 21 days could be clearly observed post IR. And the activated M/M still can be observed during the whole experiment. RESULTS: The Evans blue extravasation of BBB gradually declined from day 1 to day 21. Compared to the control group, microbubbles containing PS were taken up more by activated M/M (approximately twofold) both in vitro and in vivo. CONCLUSIONS: PS-MBs combined with ultrasound (US) exposure could safely open BBB, and the resulting PS nanoparticles (PS-NPs) could further target activated M/M in the neuroinflammation.
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Barrera Hematoencefálica/patología , Encefalitis , Hipoxia-Isquemia Encefálica/complicaciones , Microburbujas/uso terapéutico , Fosfatidilserinas/uso terapéutico , Ultrasonografía/efectos adversos , Animales , Infarto Cerebral/etiología , Infarto Cerebral/patología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Encefalitis/diagnóstico por imagen , Encefalitis/etiología , Encefalitis/patología , Macrófagos/fisiología , Imagen por Resonancia Magnética , Masculino , Microglía/fisiología , Permeabilidad , Ratas , Ratas Sprague-Dawley , Sincalida/metabolismo , Factores de TiempoRESUMEN
Insulin-like growth factor binding protein 4 (IGFBP-4) was reported to trigger cellular senescence and reduce cell growth of bone marrow mesenchymal stem cells (BMSCs), but its contribution to neurogenic differentiation of BMSCs remains unknown. In the present study, BMSCs were isolated from the femur and tibia of young rats to investigate effects of IGFBP-4 on BMSC proliferation and growth of neurospheres derived from BMSCs. Bone marrow mesenchymal stem cell proliferation was assessed using CCK-8 after treatment with IGFBP-4 or blockers of IGF-IR and ß-catenin. Phosphorylation levels of Akt, Erk, and p38 in BMSCs were analysed by Western blotting. Bone marrow mesenchymal stem cells were induced into neural lineages in NeuroCult medium; the number and the size of BMSC-derived neurospheres were counted after treatment with IGFBP-4 or the blockers. It was shown that addition of IGFBP-4 inhibited BMSC proliferation and immunodepletion of IGFBP-4 increased the proliferation. The blockade of IGF-IR with AG1024 increased BMSC proliferation and reversed IGFBP-4-induced proliferation inhibition; however, blocking of ß-catenin with FH535 did not. p-Erk was significantly decreased in IGFBP-4-treated BMSCs. IGFBP-4 promoted the growth of neurospheres derived from BMSCs, as manifested by the increases in the number and the size of the derived neurospheres. Both AG1024 and FH535 inhibited the formation of NeuroCult-induced neurospheres, but FH535 significantly inhibited the growth of neurospheres in NeuroCult medium with EGF, bFGF, and IGFBP-4. The data suggested that IGFBP-4 inhibits BMSC proliferation through IGF-IR pathway and promotes growth of BMSC-derived neurospheres via stabilizing ß-catenin.
Asunto(s)
Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteína 4 de Unión a Factor de Crecimiento Similar a la Insulina/farmacología , Animales , Células de la Médula Ósea/citología , Células Cultivadas , Factor de Crecimiento Epidérmico/farmacología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Proteína 4 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Proteína 4 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Microscopía Fluorescente , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas , Ratas Sprague-Dawley , Receptor IGF Tipo 1/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacología , Sulfonamidas/farmacología , Tirfostinos/farmacología , beta Catenina/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
OBJECTIVE: To investigate the protective effect of prostaglandin E1 (PGE-1) against brain injury induced by hyperoxia in neonatal rats and observe the changes in the expression of glucose-regulated protein 78 (GRP78) and cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), and to provide a theoretical basis for the clinical application of PGE-1 in the treatment of neonatal brain injury induced by hyperoxia. METHODS: Sixty neonatal Wistar rats were randomly divided into air control group, hyperoxic brain injury model group, and hyperoxic brain injury+PGE-1 group. All rats except those in the air control group were treated to establish a hyperoxic brain injury model. From the first day of modeling, the rats in the hyperoxia brain injury+PGE-1 group were intraperitoneally injected with PGE-1 2â µg/kg daily for 7 consecutive days, while the other two groups were treated with normal saline instead. The water content of brain tissue was measured; the pathological changes of brain tissue were evaluated by hematoxylin-eosin staining; the apoptosis of brain cells was assessed by nuclear staining combined with TUNEL staining; the protein expression of GRP78 and CHOP in brain tissue was measured by Western blot. RESULTS: The water content of brain tissue in the hyperoxic brain injury model group was significantly higher than that in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the water content of brain tissue in the hyperoxic brain injury+PGE-1 group was significantly higher than that in the air control group (P<0.05). The pathological section of brain tissue showed inflammatory cell infiltration and mild cerebrovascular edema in the brain parenchyma in the hyperoxic brain injury model group; the periparenchymal inflammation and edema in the hyperoxic brain injury+PGE-1 group were milder than those in the hyperoxic brain injury model group. The apoptosis index of brain tissue in the hyperoxic brain injury model group was significantly higher than that in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the apoptosis index of brain tissue in the hyperoxic brain injury+PGE-1 group was significantly higher than that in the air control group (P<0.05). The protein expression of GRP78 and CHOP in brain tissue was significantly higher in the hyperoxic brain injury model group than in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the protein expression of GRP78 and CHOP was significantly higher in the hyperoxic brain injury+PGE-1 group than in the air control group (P<0.05). CONCLUSIONS: PGE-1 has a protective effect against hyperoxia-induced brain injury in neonatal rats, which may be related to the inhibition of cell apoptosis by down-regulating the expression of GRP78 and CHOP.
Asunto(s)
Alprostadil/uso terapéutico , Lesiones Encefálicas/prevención & control , Hiperoxia/complicaciones , Fármacos Neuroprotectores/uso terapéutico , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Encéfalo/patología , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/patología , Proteínas de Choque Térmico/análisis , Ratas , Ratas Wistar , Factor de Transcripción CHOP/análisisRESUMEN
OBJECTIVES: Subharmonic imaging (SHI) is a technique that uses the nonlinear oscillations of microbubbles when exposed to ultrasound at high pressures transmitting at the fundamental frequency ie, fo and receiving at half the transmit frequency (ie, fo /2). Subharmonic aided pressure estimation (SHAPE) is based on the inverse relationship between the subharmonic amplitude of the microbubbles and the ambient pressure change. METHODS: Eight waveforms with different envelopes were optimized with respect to acoustic power at which the SHAPE study is most sensitive. The study was run with four input transmit cycles, first in vitro and then in vivo in three canines to select the waveform that achieved the best sensitivity for detecting changes in portal pressures using SHAPE. A Logiq 9 scanner with a 4C curvi-linear array was used to acquire 2.5 MHz radio-frequency data. Scanning was performed in dual imaging mode with B-mode imaging at 4 MHz and a SHI contrast mode transmitting at 2.5 MHz and receiving at 1.25 MHz. Sonazoid, which is a lipid stabilized gas filled bubble of perfluorobutane, was used as the contrast agent in this study. RESULTS: A linear decrease in subharmonic amplitude with increased pressure was observed for all waveforms (r from -0.77 to -0.93; P < .001) in vitro. There was a significantly higher correlation of the SHAPE gradient with changing pressures for the broadband pulses as compared to the narrowband pulses in both in vitro and in vivo results. The highest correlation was achieved with a Gaussian windowed binomial filtered square wave with an r-value of -0.95. One of the three canines was eliminated for technical reasons, while the other two produced very similar results to those obtained in vitro (r from -0.72 to -0.98; P <.01). The most consistent in vivo results were achieved with the Gaussian windowed binomial filtered square wave (r = -0.95 and -0.96). CONCLUSIONS: Using this waveform is an improvement to the existing SHAPE technique (where a square wave was used) and should make SHAPE more sensitive for noninvasively determining portal hypertension.