RESUMO
Mammalian DNA replication is initiated at numerous replication origins, which are clustered into thousands of replication domains (RDs) across the genome. However, it remains unclear whether the replication origins within each RD are activated stochastically or preferentially near certain chromatin features. To understand how DNA replication in single human cells is regulated at the sub-RD level, we directly visualized and quantitatively characterized the spatiotemporal organization, morphology, and in situ epigenetic signatures of individual replication foci (RFi) across S-phase at superresolution using stochastic optical reconstruction microscopy. Importantly, we revealed a hierarchical radial pattern of RFi propagation dynamics that reverses directionality from early to late S-phase and is diminished upon caffeine treatment or CTCF knockdown. Together with simulation and bioinformatic analyses, our findings point to a "CTCF-organized REplication Propagation" (CoREP) model, which suggests a nonrandom selection mechanism for replication activation at the sub-RD level during early S-phase, mediated by CTCF-organized chromatin structures. Collectively, these findings offer critical insights into the key involvement of local epigenetic environment in coordinating DNA replication across the genome and have broad implications for our conceptualization of the role of multiscale chromatin architecture in regulating diverse cell nuclear dynamics in space and time.
Assuntos
Fator de Ligação a CCCTC/metabolismo , Cromatina/metabolismo , Replicação do DNA , Fator de Ligação a CCCTC/genética , Cromatina/genética , Epigenômica , Humanos , Fase SRESUMO
Chronic obstructive pulmonary disease (COPD) is a chronic and devastating condition characterized by poor airflow and breath. Smoking and other environmental factors-caused inflammations triggered excessive autophagy of normal lung epithelial cells, eventually leading to impaired lung functions. Previous studies showed that ghrelin exhibited beneficial effects on patients with COPD. However, the mechanisms underlying this impact remained largely unknown. In this study, in vitro and in vivo models of COPD-associated inflammation were established, and we found that inflammation and autophagy were abonormally activated through nuclear factor kappa b (NF-κB) and activator protein-1 (AP-1) signaling pathways. Interestingly, ghrelin could inhibit the excessive inflammation pathways and autophagy induced by particle matter and/or cigarette extract in bronchial epithelial cells. Furthermore, NF-κB and AP-1 signaling were both inhibited while lung functions were significantly improved. Taken together, identification of downstream signaling of ghrelin in inflammation provided a new avenue in the treatment of COPD.
Assuntos
Autofagia/efeitos dos fármacos , Grelina/farmacologia , Doença Pulmonar Obstrutiva Crônica , Animais , Linhagem Celular , Modelos Animais de Doenças , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Camundongos , NF-kappa B/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Fator de Transcrição AP-1/metabolismoRESUMO
We have shown in the past studies that fine particulate matter (PM2.5) exposure increases airway hyperresponsiveness and leads to lung inflammation damage. Interleukin (IL)-37 plays a inhibitory role in inflammation activation and maintenance. However, the function of IL-37 in the above processes keep unclear. We aim to explore the role of IL-37 in PM2.5-induced airway hyperresponsiveness in this study. A nose-only PM2.5 online concentration, enrichment and exposure instrument was also applied to generate mice model of airway hyperresponsiveness. A transgenic mice strain using a CMV promoter to express human IL-37b (hIL-37tg) was obtained. PM2.5 exposure was shown to increase airway resistance, followed by lung inflammation and IL-1ß, TNFα, and IL-6 release, which was inhibited by IL-37tg mice and mice administrated recombinant human IL-37 intranasally (i.n). Moreover, expression of the proliferation-related protein PCNA and migration-related proteins MMP-2, MMP-9, and Vimentin was reduced in lung tissues of IL-37tg mice and mice given recombinant human IL-37 i.n. Abnormal cell contraction, proliferation, and migration of human airway smooth muscle cells (hASMCs) incubated with PM2.5 were also decreased by IL-37 treatment. In addition, IL-37 intervention of hASMCs before PM2.5 incubation decreased cytoplasmic calcium level and expression of PCNA, MMP-2, MMP-9 and Vimentin. Finally, knockdown of the IL-37 receptor IL-1R8 gene eliminated the protective effects of IL-37 in the above responses. We conclude that IL-37 inhibits inflammation activation and disease severity of airway hyperreactivity by PM2.5 induction.
Assuntos
Material Particulado , Hipersensibilidade Respiratória , Animais , Humanos , Inflamação/induzido quimicamente , Interleucina-1 , Pulmão , Camundongos , Material Particulado/toxicidade , Hipersensibilidade Respiratória/induzido quimicamente , Índice de Gravidade de DoençaRESUMO
Epidemiological research has identified that exposure to fine particulate matter (PM2.5) can increase airway hyperresponsiveness (AHR) which is considered a typical characteristic of asthma. Although the effect of PM2.5 on AHR has been elucidated to a certain degree, its exact mechanism remains unclear. Bromodomain-containing protein 4 (BRD4) is recognized as a member of the bromodomain and extraterminal (BET) family, with the ability to maintain higher-order chromatin configuration and regulate gene expression programs. The primary objective of our study was to examine the role of BRD4 in AHR triggered by PM2.5, and to elucidate its possible molecular mechanism. A mouse model with AHR was established using a nose-only PM2.5 exposure system. We observed that PM2.5 enhanced AHR in the experimental group compared to the control group, and this alteration was accompanied by increased lung inflammation and BRD4 expression in bronchi-lung tissue. However, the BRD4 inhibitor (ZL0420) could alleviate the aforementioned alterations in the mouse model with PM2.5 exposure. To explore the exact molecular mechanism, we further examined the role of BRD4 in human airway smooth muscle cells (hASMCs) after exposure to PM2.5 DMSO extracts. We found that PM2.5 DMSO extracts, which promoted the contraction and migration of hASMCs, was accompanied by an increase in the levels of BRD4, kallikrein 14 (KLK14), bradykinin 2 receptor (B2R), matrix metalloproteinases2(MMP-2), matrix metalloproteinases9(MMP-9), vimentin and bradykinin (BK) secretion, while ZL0420 and BRD4 gene silencing could reverse this response. In summary, these results demonstrate that BRD4 is an important player in AHR triggered by PM2.5, and BRD4 inhibition can ameliorate AHR induced by PM2.5. In addition, PM2.5 DMSO extracts can promote the contraction and migration of hASMCs by increasing BRD4 expression.
Assuntos
Proteínas de Ciclo Celular/metabolismo , Material Particulado/toxicidade , Hipersensibilidade Respiratória/induzido quimicamente , Fatores de Transcrição/metabolismo , Animais , Modelos Animais de Doenças , Humanos , Pulmão/efeitos dos fármacos , Camundongos , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , Pneumonia/induzido quimicamente , Hipersensibilidade Respiratória/fisiopatologiaRESUMO
Epidemiological studies have reported short-term fine particulate matter (PM2.5) exposure to increase incidence of asthma, related to the increase of airway hyperresponsiveness (AHR); however, the underlying mechanism remains unclear. Aim of this study was to elucidate the role of kallikrein in PM2.5-induced airway hyperresponsiveness and understand the underlying mechanism. Nose-only PM2.5 exposure system was used to generate a mouse model of airway hyperresponsiveness. Compared with the control group, PM2.5 exposure could significantly increase airway resistance, lung inflammation, kallikrein expression of bronchi-lung tissue and bradykinin (BK) secretion. However, these changes could be alleviated by kallikrein inhibitor. In additionï¼PM2.5 could increase the viability of human airway smooth muscle cells (hASMCs), accompanied by increased expression of kallikrein 14 (Klk14), bradykinin 2 receptor (B2R), bradykinin secretion and cytosol calcium level, while kallikrein 14 gene knockdown could significantly amelioratethe above response induced by PM2.5. Taken together, the data suggested kallikrein to play a key role in PM2.5-induced airway hyperresponsiveness, and that it could be a potential therapeutic target in asthma.
Assuntos
Poluentes Atmosféricos/toxicidade , Bradicinina/metabolismo , Calicreínas/metabolismo , Material Particulado/toxicidade , Hipersensibilidade Respiratória/induzido quimicamente , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/imunologia , Modelos Animais de Doenças , Humanos , Exposição por Inalação/efeitos adversos , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/imunologia , Miócitos de Músculo Liso/patologia , Tamanho da Partícula , Pneumonia/induzido quimicamente , Pneumonia/metabolismo , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/metabolismo , Transdução de SinaisRESUMO
BACKGROUND: Previous studies have found associations between asthma morbidity and air pollution especially in young population, (PLoS One 12:e0180522, 2017; Can J Public Health 103:4-8, 2012; Environ Health Perspect 118:449-57, 2010; Am J Respir Crit Care Med 182:307-16, 2010; J Allergy Clin Immunol 104:717-22, 2008; J Allergy Clin Immunol 104:717-22, 1999; Environ Res 111:1137-47, 2011) but most of them were conducted in areas with relatively low air pollutant level. Moreover, very few studies have investigated the effect and burden modification of heating season during which the ambient air pollution level is significantly different from that during non-heating season in north China. OBJECTIVES: This study aimed to evaluate the effect and burden modification of heating on short-term associations between adult asthma hospitalizations and ambient air pollution in the north China city of Shijiazhuang. METHODS: Generalized additive models combined with penalized distributed lag nonlinear models were used to model associations between daily asthma hospitalizations and ambient air pollutants from 1 January 2013 to 16 December 2016 in Shijiazhuang city, adjusting for long-term and seasonality trend, day of week, statutory holiday, daily mean air pressure and temperature. Attributable risks were calculated to evaluate the burden of asthma hospitalizations due to air pollutants exposure. The effect of pollutants on hospitalization and the attributable measures were estimated in heating and non-heating season separately and the comparisons between the two seasons were conducted. RESULTS: All pollutants demonstrated positive and significant impacts on asthma hospitalizations both in heating season and non-heating season, except for O3 in heating season where a negative association was observed. However, the differences of the pollutant-specific effects between the two seasons were not significant. SO2 and NO2 exposure were associated with the heaviest burden among all pollutants in heating season; meanwhile, PM10 and PM2.5 were associated with the heaviest burden in heating season. CONCLUSIONS: In conclusion, we found evidence of the effect of ambient air pollutants on asthma hospitalizations in Shijiazhuang. The central heating period could modify the effects in terms of attributable risks. The disease burden modification of heating should be taken into consideration when planning intervention measures to reduce the risk of asthma hospitalization.
Assuntos
Poluição do Ar/efeitos adversos , Asma/epidemiologia , Monitoramento Ambiental/métodos , Calefação/efeitos adversos , Hospitalização , Material Particulado/efeitos adversos , Poluentes Atmosféricos/efeitos adversos , Poluentes Atmosféricos/análise , Poluição do Ar/análise , Asma/diagnóstico , Asma/etiologia , China/epidemiologia , Feminino , Calefação/tendências , Hospitalização/tendências , Humanos , Masculino , Material Particulado/análise , Fatores de Risco , Fatores de Tempo , População Urbana/tendênciasRESUMO
Aggregation induced emission (AIE) has attracted considerable interest for the development of fluorescence probes. However, controlling the bioconjugation and cellular labeling of AIE dots is a challenging problem. Here, this study reports a general approach for preparing small and bioconjugated AIE dots for specific labeling of cellular targets. The strategy is based on the synthesis of oxetane-substituted AIEgens to generate compact and ultrastable AIE dots via photo-crosslinking. A small amount of polymer enriched with oxetane groups is cocondensed with most of the AIEgens to functionalize the nanodot surface for subsequent streptavidin bioconjugation. Due to their small sizes, good stability, and surface functionalization, the cell-surface markers and subcellular structures are specifically labeled by the AIE dot bioconjugates. Remarkably, stimulated emission depletion imaging with AIE dots is achieved for the first time, and the spatial resolution is significantly enhanced to ≈95 nm. This study provides a general approach for small functional molecules for preparing small sized and ultrastable nanodots.
Assuntos
Reagentes de Ligações Cruzadas/química , Imageamento Tridimensional , Luz , Nanopartículas/química , Nanotecnologia/métodos , Cor , Difusão Dinâmica da Luz , Molécula de Adesão da Célula Epitelial/metabolismo , Fluorescência , Humanos , Células MCF-7 , Microscopia , Microtúbulos/metabolismo , Tamanho da Partícula , Frações Subcelulares/metabolismoRESUMO
This study is aimed at improving the accuracy of oral English recognition and proposing evaluation measures with better performance. This work is based on related theories such as deep learning, speech recognition, and oral English practice. As the literature summarized, the recurrent neural network was the calculation standard, and the oral English speech recognition indicators were the main basis on which an English speech recognition model was constructed. Then, 20 English majors and 5 sets of English sentence patterns were randomly selected as the research objects. The correction standards for English oral errors were introduced into the model to achieve further improvement. The research results showed that the average concordance rate of speech recognition reached 91% through the model test. The concordance rates of words, speech, and intonation in recognition were 89%, 91%, and 86%, respectively. The model could be used as an evaluation system for English speech recognition. Therefore, the application of the deep learning scoring model in the evaluation of oral English teaching was researched in this work, which provided an effective basis for the evaluation of intelligent English teaching.
Assuntos
Aprendizado Profundo , Percepção da Fala , Idioma , Redes Neurais de Computação , FalaRESUMO
The aim of this study was to investigate the efficacy and underlying mechanism of high concentration of hydrogen on lipopolysaccharide (LPS)-induced acute lung injury (ALI). We have established a corresponding mouse model and examined the function of hydrogen inhalation on lung pathology and pulmonary edema induced by LPS, as well as contents of IL-1ß, TNF-α and IL-8. The pulmonary microvascular permeability and 66.7 % hydrogen on the expression of sirt1 and its downstream signaling molecules were tested. Results showed that 66.7 % hydrogen alleviated lung pathological changes and pulmonary edema caused by LPS, and reduced the degree of ALI by inhibiting pro-inflammatory cytokine release and oxidative stress response, thereby decreasing the expression of molecules related to intercellular adhesion. sirt1 contributed to the repair of LPS-induced ALI by hydrogen through the regulation of NF-κB and catalase expression. In conclusion, 66.7 % hydrogen protected against LPS-induced ALI by suppressing inflammatory response and oxidative stress mediated by NF-κB and catalase in a sirt1-dependent manner.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Células Endoteliais/efeitos dos fármacos , Hidrogênio/farmacologia , Inflamação/tratamento farmacológico , Pulmão/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Sirtuína 1/efeitos dos fármacos , Lesão Pulmonar Aguda/induzido quimicamente , Administração por Inalação , Animais , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Lipopolissacarídeos/farmacologia , Pulmão/irrigação sanguínea , Masculino , Camundongos , MicrovasosRESUMO
Method: Thirty-two pregnant women aged 25-35 who were hospitalized in Shanxi Maternal and Child Health Hospital from January 2019 to December 2019 were included for evaluation, including 15 normal pregnant women (NG_NO group), 6 pregnant women with GDM alone (G_NO group), and 7 pregnant women with overweight alone (NG_O group). Stools were collected from pregnant women at 24 and 37 weeks of gestation and newborns' first meconium. The v3-v4 variable region of the gut flora 16s rRNA was double-ended sequenced and bioinformatically analyzed using the Illumina MiSeq PE300 sequencing platform. Results: In the third trimester of pregnancy, there were significant differences in the composition of intestinal flora between the simple overweight group, simple GDM group, and normal pregnant group. From the second trimester to the third trimester, there was no significant change in the relative distribution of intestinal flora at the phyla classification level in normal pregnant women. The relative distribution of intestinal flora at the phylum level of newborns was significantly different from that of their mothers. The characteristic intestinal microbes of newborns in simple GDM group were g_Diaphorobacter, while the simple recombinant neonates were Nocardiaceae (f_Nocardioidaceae). In addition, the results showed significant differences in intestinal flora among the normal pregnant women group, simple GDM group, simple overweight group, and GDM overweight group. The results of ß diversity analysis showed a significant difference in intestinal microflora species composition structure between the simple overweight group and the normal pregnant group in the second trimester of pregnancy. The species composition structure of intestinal flora was similar between the simple GDM group and the normal pregnant group. In the third trimester of pregnancy, there was no significant difference in the ß diversity index among the groups, and the composition and structure of intestinal flora were similar. There were significant differences in the composition structure (ß diversity) of intestinal flora between pregnant women and their newborns in each group (P < 0.05). Correlation analysis showed that the blood glucose values of oral glucose tolerance test (OGTT)_1 h and OGTT_2 h were positively correlated with Bacteroides (Bacteroides) and negatively correlated with Proteus (Prevotella), prepregnancy BMI was negatively correlated with Bacteroides, and weight gain during pregnancy was negatively correlated with Vibrio (Desulfovibrio) in Proteus. The birth weight of newborns was positively correlated with Actinomycetes (Actinomyces), Bacteroides (Faecalibacterium), and microbacilli (Dialister) and negatively correlated with Rolston (Ralstonia). Conclusion: Gut microbiota is strongly linked to obesity and gestational diabetes.
Assuntos
Diabetes Gestacional , Microbioma Gastrointestinal , Glicemia , Índice de Massa Corporal , Criança , Feminino , Humanos , Recém-Nascido , Obesidade , Sobrepeso , Gravidez , RNA Ribossômico 16S/genéticaRESUMO
OBJECTIVE: To explore the roles of macrophages' miR-217-5p in the process of PM2.5 induced acute lung injury. METHODS: GEO database and KEGG pathway enrichment analysis as well as GSEA were used to predicted the miRNA and associated target signals. And then mice and RAW246.7 macrophages treated with PM2.5 to imitate PM2.5 induced acute lung injury environment and then transfected with miR-217-5p NC or miR-217-5p mimic. The levels of inflammatory factors TNF-α and anti-inflammatory factor IL-10 of mice serum were tested by ELISA. And the pathological changes and ROS level of mouse lung tissues were stained by HE and DHE staining. The proteins expression of phosphorylated-STAT1, total-STAT1, TNF-α, IFN-γ as well as p47, gp91, NOX4 in mice or RAW264.7 cells were tested by western blot or immunofluorescence of RAW264.7 cell slides. RESULTS: The results of bioinformatics analysis indicated the miR-217 as well as STAT1 were involved PM2.5 associated lung injury. After exposure to PM2.5, the decreased levels of serum TNF-α but not IL-10, consistent with reduced macrophages' accumulation as well as decreased ROS levels in lung tissues in miR-217-5p mimic group vs miR-217-5p NC group mice, and moreover, the protein expression levels of phosphorylated--STAT1, total-STAT1, TNF-α, IFN-γ, p47, gp91 and NOX4 in mouse lung tissues and RTAW246.7 macrophages cells were all significantly reduced with miR-217-5p mimic administration. The above phenomena were reversed by specific STAT1-inhibitor HY-N8107. CONCLUSIONS: miR-217-5p suppressed the activated STAT1-signal induced inflammation and oxidative stress trigged by PM2.5 in macrophages and resulted in the decreased lung injure caused by PM2.5.
Assuntos
Lesão Pulmonar Aguda , MicroRNAs , Lesão Pulmonar Aguda/metabolismo , Animais , Inflamação/metabolismo , Pulmão/metabolismo , Macrófagos/metabolismo , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Smog , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Airway hyperresponsiveness (AHR) is characterized by excessive bronchoconstriction in response to nonspecific stimuli, thereby leading to airway stenosis and increased airway resistance. AHR is recognized as a key characteristic of asthma and is associated with significant morbidity. At present, many studies on the molecular mechanisms of AHR have mainly focused on the imbalance in Th1/Th2 cell function and the abnormal contraction of airway smooth muscle cells. However, the specific mechanisms of AHR remain unclear and need to be systematically elaborated. In addition, the effect of air pollution on the respiratory system has become a worldwide concern. To date, numerous studies have indicated that certain concentrations of fine particulate matter (PM2.5) can increase airway responsiveness and induce acute exacerbation of asthma. Of note, the concentration of PM2.5 does correlate with the degree of AHR. Numerous studies exploring the toxicity of PM2.5 have mainly focused on the inflammatory response, oxidative stress, genotoxicity, apoptosis, autophagy, and so on. However, there have been few reviews systematically elaborating the molecular mechanisms by which PM2.5 induces AHR. The present review separately sheds light on the underlying molecular mechanisms of AHR and PM2.5-induced AHR.
Assuntos
Poluição do Ar , Asma , Hipersensibilidade Respiratória , Poluição do Ar/efeitos adversos , Humanos , Pulmão , Material Particulado/toxicidadeRESUMO
Background: With the control of the epidemic, adolescents' mental outlook might have improved. However, little evidence existed with regard to the psychological status of adolescents in post-COVID-19 era. This present study aimed to explore the psychological status of high school students after the epidemic getting eased. Methods: A web-based cross-sectional survey was used to obtain data from three high schools, including the demographic information, the Patient Health Questionnaire-9 (PHQ-9), the Generalized Anxiety Disorder-7 (GAD-7), the Self-Rating Scale of Sleep (SRSS), and self-designed general recent-status questionnaire. Correlation analysis was performed to explore potential associations between the depression symptoms, anxiety symptoms, and sleep status. The PHQ-9 and GAD-7 differences between nowadays data and the data enrolled 12 months before were also compared. Result: A total of 1,108 qualified questionnaires were obtained. The prevalence of depressive and anxious symptoms was 27.5 and 21.3%, respectively, from mild to severe in all students, while 11.8% of these high students got sleep disturbances. Both the rate and the severity of depression, anxiety and sleep problems of female students were higher than male students. Grade three students suffered higher prevalence and severer mental disturbances than the other two grades. There were significant correlations between the depression symptoms, anxiety symptoms, and sleep status. The psychological status has been improved in nowadays high school students compared with the sample enrolled 12 months before. Conclusion: As a supplement to our former study, this present research provided a perspective on the psychological status of high school students 1 year after the COVID-19 pandemic being well controlled. We should pay attention to the psychological status of high school students, and should also notice the progresses made by this special group after the epidemic.
RESUMO
HLA-F, a nonclassical HLA class I molecule, is required for regulating immune tolerance. In recent years, HLA-F has been found to play a role in a variety of cancers, including glioma (GM). Additionally, high expression of HLA-F predicts the poor overall survival of individuals with GM. However, the functions of HLA-F in GM remain to be further elucidated. In this study, we found that HLA-F expression was elevated in GM tissues. High levels of HLA-F resulted in a high cell proliferation index and predicted GM recurrence. Forced expression of HLA-F promoted the growth of murine C8-D1A cells transplanted in immunodeficient Rag2-/- mice. In contrast, silencing HLA-F inhibited cell growth in vitro. Furthermore, targeting HLA-F with an anti-HLA-F antibody suppressed the growth of C8-D1A cells stably expressing HLA-F transplanted in immunodeficient Rag2-/- mice. In further experiments, we found that forced expression of HLA-F contributed to the aerobic glycolysis phenotype in C8-D1A cells along with an increase in HK2 protein stabilization. Conversely, silencing HK2 by shRNA reduced HLA-F-mediated glycolysis and cell proliferation. Our data indicated that HLA-F promoted cell proliferation via HK2-dependent glycolysis. HLA-F could be a potential therapeutic target for the treatment of GM.
Assuntos
Glioma , Glicólise/fisiologia , Hexoquinase/metabolismo , Antígenos de Histocompatibilidade Classe I , Animais , Linhagem Celular Tumoral , Proliferação de Células/genética , Descoberta de Drogas , Regulação Neoplásica da Expressão Gênica , Inativação Gênica/fisiologia , Marcação de Genes/métodos , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , CamundongosRESUMO
Expansion microscopy (ExM) provides nanoscale resolution on conventional microscopes via physically enlarging specimens with swellable polyelectrolyte gels. However, challenges involving fluorophore degradation and dilution during sample expansion have yet to be overcome. Herein, sequential cellular targeting, gel anchoring, and high-fidelity fluorescence reported using multifunctional polymer dots (Pdots) designed for ExM applications are demonstrated. The impressive brightness of the Pdots facilitates multicolor ExM, thereby enabling visualization of a variety of subcellular structures and neuron synapses. The average fluorescence intensities of Pdots in ExM range from ≈3 to 6 times higher than those achieved using commercially available Alexa dyes. Moreover, the fluorescence brightness and optical fluctuation are significantly improved by a surfactant-containing expansion buffer, which enables further resolution enhancement via super-resolution optical fluctuation imaging (SOFI). The combination of ExM and SOFI allows subcellular structures of ≈30 nm to be resolved by conventional microscopes. These results highlight the immense potential of multifunctional Pdots for ExM-enhanced super-resolution imaging.
RESUMO
Few researches have been investigated on the effects of ambient air pollutants from coal combustion on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) hospitalizations. The whole time series was split into heating season and non-heating season. We used a quasi-Poisson generalized linear regression model combined with distributed lag non-linear models (DLNMs) to estimate the relative cumulative risk and calculate the air pollutant hospitalization burden of AECOPD for lag 0-7 days in heating season and non-heating season. There were higher PM2.5, PM10, NO2, SO2, and CO concentrations in heating seasons than non-heating season in Shijiazhuang; however, O3 was higher in non-heating season than heating season. The AECOPD-associated relative cumulative risks for PM2.5, PM10, NO2, and SO2 for lag 0-7 days were significantly positively associated with hospitalization in heating and non-heating season; we found that the cumulative relative risk of NO2 was the greatest in every 1 unit of air pollutants during the heating season and the cumulative relative risk of SO2 was the greatest during the non-heating season. The results showed that 17.8%, 12.9%, 1.7%, 16.7%, and 10.5% of AECOPD hospitalizations could be attributable to PM2.5, PM10, SO2, NO2, and CO exposure in heating season, respectively. However, the results showed that 19.5%, 22.4%, 15%, 8.3%, and 10.4% of AECOPD hospitalizations could be attributable to PM2.5, PM10, SO2, NO2, and O3 exposure in non-heating season, respectively. The attributable burden of AECOPD hospitalization in heating season and non-heating season are different. PM2.5, PM10, NO2, and CO are the main factors of heating season, while PM10, PM2.5, SO2, and O3 are the main factors of non-heating season. In conclusions, the centralized heating can change the influence of attributable risk. When government departments formulate interventions to reduce the risk of acute hospitalization of chronic obstructive pulmonary disease (COPD), the influence of heating on disease burden should be considered.
Assuntos
Poluição do Ar/análise , Hospitalização/estatística & dados numéricos , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Poluição do Ar/efeitos adversos , China/epidemiologia , Calefação , Humanos , Doença Pulmonar Obstrutiva Crônica/etiologia , Fatores de Risco , Estações do AnoRESUMO
Few studies have investigated the acute exacerbations of chronic obstructive pulmonary disease (AECOPD)-associated attributable burden under exposure to high levels of air pollution among Asians. Data on hospitalization for AECOPD, air pollution and meteorological factors from 1 January 2013 to 31 December 2016 were collected in Shijiazhuang, China. We used a Poisson generalized linear regression model combined with a distributed lag nonlinear model (DLNM) to evaluate the relative cumulative risk for a lag of 0-7 days and examined the potential effect modifications by age and sex via stratification analyses, controlling for long-term trends, seasonal patterns, meteorological factors, and other possible confounders. Then, we computed hospitalization percentages attributable to air pollutants. The AECOPD-associated relative cumulative risks for PM2.5, PM10, NO2, SO2, and CO for a lag of 0-7 days were significantly positively correlated with hospitalization. The associations were stronger in females and retired patients. The NO2 Cum RR of AECOPD admission was the greatest. A 10µg/m3 increase in daily NO2 concentration was associated with 6.7% and 5.7% increases in COPD hospitalizations in the retired and female groups, respectively. The results showed that 13%, 9.4%, 1.7%, 9.7%, and 8.8% of AECOPD hospitalizations were attributable to exposure to PM2.5, PM10, SO2, NO2, and CO, respectively. If the air pollutant concentration was reduced to the 24-h average grade II levels of NAAQS of China, the AECOPD attributable percentage for PM2.5 and PM10 would decrease by 80%. The air pollutants PM2.5, PM10, SO2, NO2, and CO were significantly relevant to AECOPD-associated hospitalization. The associations differed by individual characteristics. The retired and female populations were highly vulnerable.
Assuntos
Poluição do Ar/efeitos adversos , Hospitalização/estatística & dados numéricos , Material Particulado/efeitos adversos , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Doença Pulmonar Obstrutiva Crônica/etiologia , Idoso , Poluentes Atmosféricos/efeitos adversos , Poluentes Atmosféricos/análise , Poluição do Ar/análise , China/epidemiologia , Exposição Ambiental/efeitos adversos , Feminino , Humanos , Masculino , Conceitos Meteorológicos , Pessoa de Meia-Idade , Material Particulado/análiseRESUMO
BACKGROUND: Ambient fine particulate matter (PM2.5) could induce lung injury. Aryl hydrocarbon receptor (AhR) is involved in the molecular mechanisms of prooxidative and pro-inflammatory effect of PM2.5. Molecular hydrogen has antioxidant properties. The protective effect and mechanism of hydrogen on PM2.5-induced lung injury remain unclear. OBJECTIVES: This study aimed to determine whether hydrogen could alleviate lung injury in a rat model of subacute exposure to concentrated ambient PM2.5, and explore the mechanism related to AhR. METHODS: Male Wastar rats were exposed to either concentrated ambient particles (CAPs) (diameter: ≤2.5⯵m, average concentration: 1328⯱â¯730⯵g/m3) or filtered air (FA) by nose-only inhalation (5â¯h/day, 5â¯days/week for 4â¯weeks). Hydrogen-treated rats inhaled 66.7% hydrogen from water electrolysis for 2â¯h after each exposure to CAPs or FA. RESULTS: CAPs inhalation induced lung injury, as demonstrated by pulmonary function decrease, histopathological damage, mucus hypersecretion [Periodic acid-Schiff (PAS) staining for mucins, immunohistochemistry and quantitative real-time PCR (RT-qPCR) for mucin 5AC (MUC5AC) expression], increased pro-inflammatory cytokines (TNF-α, IL-8 and IL-1ß) and oxidative damage indexes [malondialdehyde (MDA) and 8-isoprostane F2α (8-iso-PG)]. While, hydrogen inhalation significantly alleviated the damages mentioned above. In addition, low expression of AhR in lung tissues determined by Western Blot was found after CAPs exposure, whereas hydrogen inhibited AhR decline induced by CAPs. CONCLUSIONS: High concentrations of hydrogen could ameliorate pulmonary dysfunction, airway mucus hypersecretion, oxidation damage, and inflammation response in rats exposed to concentrated ambient PM2.5. Additionally, hydrogen alleviates lung injury induced by PM2.5 possibly through AhR-dependent mechanisms.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Hidrogênio/farmacologia , Lesão Pulmonar/tratamento farmacológico , Lesão Pulmonar/metabolismo , Material Particulado/efeitos adversos , Receptores de Hidrocarboneto Arílico/metabolismo , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Inflamação/metabolismo , Lesão Pulmonar/induzido quimicamente , Masculino , Tamanho da Partícula , Ratos , Ratos WistarRESUMO
OBJECTIVES: The present study was aimed to examine the effects of chlorpromazine on sleep quality, clinical and emotional measures in people suffering from schizophrenia. PATIENTS AND METHODS: Twenty-five patients with schizophrenia or schizoaffective disorder were enrolled in this study. Our study included a one-week running-in no-treatment period and two-month experimental period. Patients received chlorpromazine during the experimental period. The baseline and treatment outcome were recorded. The objective and subjective sleep were respectively measured by a wrist actigraph and two sleep questionnaires (Mini Sleep Questionnaire (MSQ) and Pittsburgh Sleep Quality Index Hebrew Translation (PSQI-H)). Besides, Brief Psychiatric Rating Scale (BPRS) and Positive and Negative Syndrome Scale (PANSS) were performed to assess the clinical psychopathology levels, and while Calgary Depression Scale for schizophrenia (CDSS) and Hamilton Anxiety Rating Scale (HAS) had been carried out to examine the emotional changes. RESULTS: There was a significant effect of chlorpromazine treatment on four objective sleep variables: longest wake episode, sleep onset latency, sleep percentage, and mean activity level (all Pâ¯<â¯0.05). However, no significant differences were found in the subjective sleep measures. Likewise, psychopathology levels and emotional measures (depression level and anxiety level) were statistically improved by chlorpromazine treatment compared to the baseline (Pâ¯<â¯0.05 or Pâ¯<â¯0.01). CONCLUSION: Our results demonstrate that chlorpromazine could improve the insomnia and psychopathology symptoms of patients with schizophrenia.
Assuntos
Antipsicóticos/uso terapêutico , Clorpromazina/uso terapêutico , Esquizofrenia/complicações , Esquizofrenia/tratamento farmacológico , Distúrbios do Início e da Manutenção do Sono/tratamento farmacológico , Distúrbios do Início e da Manutenção do Sono/etiologia , Actigrafia , Adulto , Idoso , Emoções , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Escalas de Graduação Psiquiátrica , Transtornos Psicóticos/complicações , Transtornos Psicóticos/tratamento farmacológico , Transtornos Psicóticos/psicologia , Psicologia do Esquizofrênico , Resultado do Tratamento , Adulto JovemRESUMO
The advance of optical super-resolution fluorescence microscopy has revolutionized our vision of the subcellular world. Further improvement in the spatial resolution is of great significance for structural and functional investigations. The recently developed expansion microscopy (ExM), which achieves sub-diffraction imaging via physical expansion of the sample, provides a great opportunity for further resolution enhancement of existing optical super-resolution techniques. However, although such combination seems apparent, several technical obstacles, especially the dramatic loss of fluorescence signal during ExM sample preparation, have hampered this goal. In this work, aiming at this challenge, we have developed new strategies to retain and increase the fluorescence of the expanded sample. With the new labeling methods, we have successfully made the labeling density of expanded samples sufficing the Nyquist sampling criteria for optical super-resolution imaging, such as stimulated emission depletion microscopy (STED) and super-resolution optical fluctuation imaging (SOFI). The newly developed expansion nanoscopic imaging (ExN) approaches, i.e. ExSTED and ExSOFI, demonstrated up to 4-fold resolution enhancement compared to standard STED and SOFI, providing a simple and effective way to realize high resolution imaging both at the cellular and tissue level.