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The regulation of mammalian reproductive activity is tightly dependent on the HPG axis crosstalk, in which several reproductive hormones play important roles. Among them, the physiological functions of gonadotropins are gradually being uncovered. However, the mechanisms by which GnRH regulates FSH synthesis and secretion still need to be more extensively and deeply explored. With the gradual completion of the human genome project, proteomes have become extremely important in the fields of human disease and biological process research. To explore the changes of protein and protein phosphorylation modifications in the adenohypophysis after GnRH stimulation, proteomics and phosphoproteomics analyses of rat adenohypophysis after GnRH treatment were performed by using TMT markers, HPLC classification, LC/MS, and bioinformatics analysis in this study. A total of 6762 proteins and 15,379 phosphorylation sites contained quantitative information. Twenty-eight upregulated proteins and fifty-three downregulated proteins were obtained in the rat adenohypophysis after GnRH treatment. The 323 upregulated phosphorylation sites and 677 downregulated phosphorylation sites found in the phosphoproteomics implied that a large number of phosphorylation modifications were regulated by GnRH and were involved in FSH synthesis and secretion. These data constitute a protein-protein phosphorylation map in the regulatory mechanism of "GnRH-FSH," which provides a basis for future studies on the complex molecular mechanisms of FSH synthesis and secretion. The results will be helpful for understanding the role of GnRH in the development and reproduction regulated by the pituitary proteome in mammals.
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Hormona Liberadora de Gonadotropina , Adenohipófisis , Animales , Ratas , Hormona Folículo Estimulante/metabolismo , Hormona Liberadora de Gonadotropina/metabolismo , Hormona Luteinizante/metabolismo , Adenohipófisis/metabolismo , ProteómicaRESUMEN
Prolactin (PRL) is an important hormone that is secreted by the pituitary gland and plays an important role in the growth, development and reproduction of organisms. Thyrotropin-releasing hormone (TRH) is a common prolactin-releasing factor that regulates the synthesis and secretion of prolactin. In recent studies, microRNAs (miRNAs) have been found to play a key role in the regulation of pituitary hormones. However, there is a lack of systematic studies on the regulatory role that TRH plays on the pituitary transcriptome, and the role of miRNAs in the regulation of PRL synthesis and secretion by TRH lacks experimental evidence. In this study, we first investigated the changes in PRL synthesis and secretion in the rat pituitary gland after TRH administration. The results of transcriptomic analysis after TRH treatment showed that 102 genes, including those that encode Nppc, Fgf1, PRL, Cd63, Npw, and Il23a, were upregulated, and 488 genes, including those that encode Lats1, Cacna2d1, Top2a, and Tfap2a, were downregulated. These genes are all involved in the regulation of prolactin expression. The gene expression of miR-126a-5p, which regulates the level of PRL in the pituitary gland, was screened by analysis prediction software and by a dual luciferase reporter system. The data presented in this study demonstrate that TRH can regulate prolactin synthesis and secretion through miR-126a-5p, thereby improving our understanding of the molecular mechanism of TRH-mediated PRL secretion and providing a theoretical basis for the role of miRNAs in regulating the secretion of pituitary hormones.
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MicroARNs , Adenohipófisis , Animales , Ratas , MicroARNs/genética , MicroARNs/metabolismo , Adenohipófisis/metabolismo , Hormonas Hipofisarias/metabolismo , Prolactina/genética , Prolactina/metabolismo , Hormona Liberadora de Tirotropina/genética , Hormona Liberadora de Tirotropina/metabolismoRESUMEN
BACKGROUND: With the widespread use of rifampicin and isoniazid, bacterial resistance has become a growing problem. Additionally, the lack of relevant baseline information for the frequency of drug-resistant tuberculosis (TB) gene mutations is a critical issue, and the incidence of this infection in the city of Changchun has not investigated to date. However, compared with the slow traditional methods of drug susceptibility testing, recently developed detection methods, such as rifampicin and isoniazid resistance-related gene chip techniques, allow for rapid, easy detection and simultaneous testing for mutation frequency and drug resistance. METHODS: In this study, the rifampicin and isoniazid resistance-related gene mutation chip method was employed for an epidemiological investigation. To assess the gene mutation characteristics of drug-resistant TB and evaluate the chip method, we tested 2143 clinical specimens from patients from the infectious diseases hospital of Changchun city from January to December 2016. The drug sensitivity test method was used as the reference standard. RESULTS: The following mutation frequencies of sites in the rifampicin resistance gene rpoB were found: Ser531Leu (52.6%), His526Tyr (12.3%), and Leu511Pro (8.8%). The multidrug-resistance (MDR)-TB mutation frequency was 34.7% for rpoB Ser531Leu and katG Ser315Thr, 26.4% for rpoB Ser531Leu and inhA promoter - 15 (C â T), and 10.7% for rpoB His526Tyr and katG Ser315Thr. In addition, drug susceptibility testing served as a reference standard. In previously treated clinical cases, the sensitivity and specificity of GeneChip were 83.1 and 98.7% for rifampicin resistance, 79.9 and 99.6% for isoniazid resistance, and 74.1 and 99.8% for MDR-TB. CONCLUSIONS: Our experimental results show that the chip method is accurate and reliable; it can be used to detect the type of drug-resistant gene mutation in clinical specimens. Moreover, this study can be used as a reference for future research on TB resistance baselines.
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Antituberculosos/uso terapéutico , Mycobacterium tuberculosis/genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Tuberculosis/tratamiento farmacológico , Antituberculosos/farmacología , Proteínas Bacterianas/genética , China , Farmacorresistencia Bacteriana/efectos de los fármacos , Farmacorresistencia Bacteriana/genética , Frecuencia de los Genes , Humanos , Pruebas de Sensibilidad Microbiana , Mutación , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/aislamiento & purificación , Rifampin/farmacología , Rifampin/uso terapéutico , Tuberculosis/microbiología , Tuberculosis/patologíaRESUMEN
Neuroinflammation plays a very important role in the pathogenesis of Parkinson's disease (PD). After activation, microglia produce pro-inflammatory mediators that damage surrounding neurons. Consequently, the inhibition of microglial activation might represent a new therapeutic approach of PD. Vanillin has been shown to protect dopaminergic neurons, but the mechanism is still unclear. Herein, we further study the underlying mechanisms in lipopolysaccharide (LPS)-induced PD models. In vivo, we firstly established rat models of PD by unilateral injection of LPS into substantia nigra (SN), and then examined the role of vanillin in motor dysfunction, microglial activation and degeneration of dopaminergic neurons. In vitro, murine microglial BV-2 cells were treated with vanillin prior to the incubation of LPS, and then the inflammatory responses and the related signaling pathways were analyzed. The in vivo results showed that vanillin markedly improved the motor dysfunction, suppressed degeneration of dopaminergic neurons and inhibited microglial over-activation induced by LPS intranigral injection. The in vitro studies demonstrated that vanillin reduces LPS-induced expression of inducible nitric oxide (iNOS), cyclooxygenase-2 (COX-2), IL-1ß, and IL-6 through regulating ERK1/2, p38 and NF-κB signaling. Collectively, these data indicated that vanillin has a role in protecting dopaminergic neurons via inhibiting inflammatory activation.
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Benzaldehídos/farmacología , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/metabolismo , Inflamación/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Fármacos Neuroprotectores/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Muerte Celular , Línea Celular , Citocinas/metabolismo , Neuronas Dopaminérgicas/patología , Inflamación/patología , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/metabolismo , Masculino , Microglía/efectos de los fármacos , Microglía/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Actividad Motora/efectos de los fármacos , Ratas , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
The pathogenesis of Parkinson's disease (PD) often involves the over-activation of microglia. Over-activated microglia could produce several inflammatory mediators, which trigger excessive inflammation and ultimately cause dopaminergic neuron damage. Anti-inflammatory effects of glucagon-like peptide-2 (GLP-2) in the periphery have been shown. Nonetheless, it has not been illustrated in the brain. Thus, in this study, we aimed to understand the role of GLP-2 in microglia activation and to elucidate the underlying mechanisms. BV-2 cells were pretreated with GLP-2 and then stimulated by lipopolysaccharide (LPS). Cells were assessed for the responses of pro-inflammatory enzymes (iNOS and COX-2) and pro-inflammatory cytokines (IL-1ß, IL-6 and TNF-α); the related signaling pathways were evaluated by Western blotting. The rescue effect of GLP-2 on microglia-mediated neurotoxicity was also examined. The results showed that GLP-2 significantly reduced LPS-induced production of inducible nitric oxide synthase (iNOS), cyclooxygenase-s (COX-2), IL-1ß, IL-6 and TNF-α. Blocking of Gαs by NF449 resulted in a loss of this anti-inflammatory effect in BV-2 cells. Analyses in signaling pathways demonstrated that GLP-2 reduced LPS-induced phosphorylation of ERK1/2, JNK1/2 and p65, while no effect was observed on p38 phosphorylation. In addition, GLP-2 could suppress microglia-mediated neurotoxicity. All results imply that GLP-2 inhibits LPS-induced microglia activation by collectively regulating ERK1/2, JNK1/2 and p65.
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Péptido 2 Similar al Glucagón/metabolismo , Inflamación/metabolismo , Transducción de Señal , Animales , Línea Celular Transformada , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Citocinas/genética , Citocinas/metabolismo , Subunidades alfa de la Proteína de Unión al GTP Gs/metabolismo , Expresión Génica , Péptido 2 Similar al Glucagón/farmacología , Inflamación/inmunología , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/inmunología , Sistema de Señalización de MAP Quinasas , Microglía/metabolismo , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Proteína Quinasa 9 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosforilación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Inflammatory bowel disease (IBD) is difficult to cure, and formulating a dietary plan is an effective means to prevent and treat this disease. Wheat peptide contains a variety of bioactive peptides with anti-inflammatory and antioxidant functions. The results of this study showed that preventive supplementation with wheat peptide (WP) can significantly alleviate the symptoms of dextran sulfate sodium (DSS)-induced colitis in mice. WP can increase body weight, alleviate colon shortening, and reduce disease activity index (DAI) scores. In addition, WP improved intestinal microbial disorders in mice with colitis. Based on LC-MS, a total of 313 peptides were identified in WP, 4 of which were predicted to be bioactive peptides. The regulatory effects of WP and four bioactive peptides on the Keap1-Nrf2 signaling pathway were verified in Caco-2 cells. In conclusion, this study demonstrated that WP alleviates DSS-induced colitis by helping maintain gut barrier integrity and targeting the Keap1-Nrf2 axis; these results provided a rationale for adding WP to dietary strategies to prevent IBD.
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Colitis , Sulfato de Dextran , Proteína 1 Asociada A ECH Tipo Kelch , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2 , Péptidos , Transducción de Señal , Triticum , Animales , Humanos , Masculino , Ratones , Células CACO-2 , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/metabolismo , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Mucosa Intestinal/metabolismo , Mucosa Intestinal/efectos de los fármacos , Proteína 1 Asociada A ECH Tipo Kelch/efectos de los fármacos , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Factor 2 Relacionado con NF-E2/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Péptidos/farmacología , Transducción de Señal/efectos de los fármacos , Triticum/químicaRESUMEN
The incidence of ulcerative colitis (UC) is increasing annually, and UC has a serious impact on patients' lives. Polysaccharides have gained attention as potential drug candidates for treating ulcerative colitis (UC) in recent years. Huaier (Trametes robiniophila Murr) is a fungus that has been used clinically for more than 1000 years, and its bioactive polysaccharide components have been reported to possess immunomodulatory effects, antitumour potential, and renoprotective effects. In this study, we aimed to examine the protective effects and mechanisms of Huaier polysaccharide (HP) against UC. Based on the H2O2-induced oxidative stress model in HT-29 cells and the dextran sulphate sodium salt (DSS)-induced UC model, we demonstrated that Huaier polysaccharides significantly alleviated DSS-induced colitis (weight loss, elevated disease activity index (DAI) scores, and colonic shortening). In addition, HP inhibited oxidative stress and inflammation and alleviated DSS-induced intestinal barrier damage. It also significantly promoted the expression of the mucin Muc2. Furthermore, HP reduced the abundance of harmful bacteria Escherichia-Shigella and promoted the abundance of beneficial bacteria Muribaculaceae_unclassified, Anaerotruncus, and Ruminococcaceae_unclassified to regulate the intestinal flora disturbance caused by DSS. Nontargeted metabolomics revealed that HP intervention would modulate metabolism by promoting levels of 3-hydroxybutyric acid, phosphatidylcholine (PC), and phosphatidylethanolamine (PE). These results demonstrated that HP had the ability to mitigate DSS-induced UC by suppressing oxidative stress and inflammation, maintaining the intestinal barrier, and modulating the intestinal flora. These findings will expand our knowledge of how HP functions and offer a theoretical foundation for using HP as a potential prebiotic to prevent UC.
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Sulfato de Dextran , Microbioma Gastrointestinal , Estrés Oxidativo , Polisacáridos , Microbioma Gastrointestinal/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Animales , Humanos , Polisacáridos/farmacología , Ratones , Masculino , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/microbiología , Modelos Animales de Enfermedad , Inflamación/tratamiento farmacológico , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Células HT29 , Ratones Endogámicos C57BL , Colitis/inducido químicamente , Colitis/tratamiento farmacológicoRESUMEN
Mastitis affects the milk quality and yield and is the most expensive disease in dairy cows. Elucidation of the pathogenesis of mastitis is of great importance for disease control. As a medium of intercellular communication, exosomes play key roles in various inflammatory diseases by regulating macrophage polarization. However, the molecular factors in exosomes that mediate the intercellular communication between mammary epithelial cells and macrophages during mastitis remain to be further explored. In this study, we isolated and identified mammary epithelial cell-derived exosomes from a lipopolysaccharide (LPS)/lipoteichoic acid (LTA)-induced mastitis cell model, and we demonstrated that exosomes from LPS/LTA-stimulated mammary epithelial cells promote M1-type macrophage polarization in vivo and in vitro. Based on the results of high-throughput sequencing, we constructed a differential miRNA (microRNA) expression profile of exosomes and demonstrated that miR-221-3p was highly expressed. Furthermore, in vivo and in vitro experiments, combined with coculture experiments and fluorescence tracing, showed that high miR-221-3p expression promoted M1-type macrophage polarization, demonstrating the transcellular role of miR-221-3p. Mechanistically, dual luciferase reporter gene assays and rescue assays showed that miR-221-3p regulated macrophage polarization by targeting Igf2bp2. The results of this study will deepen our understanding of the pathogenesis of mastitis, and the molecular regulatory axis that was established in this study is expected to be a target for mastitis treatment.
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Ulcerative colitis (UC) is a chronic noninfectious intestinal disease that severely affects patients' quality of life. Agaricus blazei Murrill polysaccharide (ABP) is an effective active ingredient extracted from Agaricus blazei Murrill (ABM). It has good efficacy in inhibiting tumor cell growth, lowering blood pressure, and improving atherosclerosis. However, its effect on colitis is unclear. The aim of this study was to analyze the protective effects and potential mechanisms of ABP against dextran sulfate sodium (DSS)-induced acute colitis in mice. The results showed that dietary supplementation with ABP significantly alleviated DSS-induced colitis symptoms, inflammatory responses, and oxidative stress. Meanwhile, ABP intervention was able to maintain the integrity of the intestinal mechanical barrier by promoting the expression of ZO-1 and Occludin tight junction proteins and facilitating mucus secretion. Moreover, 16S rRNA sequencing results suggested that ABP intervention was able to alleviate DSS-induced gut microbiota disruption, and nontargeted metabolomics results indicated that ABP was able to remodel metabolism. In conclusion, these results demonstrate that dietary supplementation with ABP alleviated DSS-induced acute colitis by maintaining intestinal barrier integrity and remodeling metabolism. These results improve our understanding of ABP function and provide a theoretical basis for the use of dietary supplementation with ABP for the prevention of ulcerative colitis.
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Colitis Ulcerosa , Colitis , Enfermedades no Transmisibles , Humanos , Animales , Ratones , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Calidad de Vida , ARN Ribosómico 16S , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Polisacáridos/farmacología , Sulfato de Dextran , Ratones Endogámicos C57BL , Modelos Animales de Enfermedad , ColonRESUMEN
Hair loss disorders such as androgenetic alopecia have caused serious disturbances to normal human life. Animal models play an important role in exploring pathogenesis of disease and evaluating new therapies. NIH hairless mice are a spontaneous hairless mouse discovered and bred in our laboratory. In this study, we resequenced the genomes of NIH normal mice and NIH hairless mice and obtained 3,575,560 high-quality, plausible SNP loci and 995,475 InDels. The Euclidean distance algorithm was used to assess the association of SNP loci with the hairless phenotype, at a threshold of 0.62. Two regions of chromosome 18 having the highest association with the phenotype contained 345 genes with a total length of 13.98 Mb. The same algorithm was used to assess the association of InDels with the hairless phenotype at a threshold of 0.54 and revealed a region of 25.45 Mb in length, containing 518 genes. The mutation candidate gene Lama3 (NM_010680.2: c.652C>T; NP_034810.1: p. Arg217Cys) was selected based on the results of functional gene analysis and mutation prediction screening. Lama3 (R217C) mutant mice were further constructed using CRISPR/Cas9 technology, and the relationship between Lama3 point mutations and the hairless phenotype were clarified by phenotypic observation. The results showed that male Lama3 point mutation mice started to lose hair on the 80th day after birth, and the hair loss area gradually expanded over time. H&E staining of skin sections showed that the point mutation mice had increased sebaceous glands in the dermis and missing hair follicle structure (i.e., typical symptoms of androgenetic alopecia). This study is a good extension of the current body of knowledge about the function of Lama3, and the constructed Lama3 (R217C) mutant mice may be a good animal model for studying androgenetic alopecia.
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Andrógenos , Laminina , Mutación Missense , Animales , Masculino , Ratones , Alopecia/genética , Alopecia/patología , Proteínas de la Matriz Extracelular/genética , Cabello/patología , Ratones Pelados , Mutación , Laminina/genéticaRESUMEN
The prevalence of inflammatory bowel disease (IBD) is progressively rising each year, emphasizing the significance of implementing rational dietary interventions for disease prevention. Oats, being a staple agricultural product, are abundant in protein content. This study aimed to investigate the protective effects and underlying mechanisms of oat peptides (OPs) in a mouse model of acute colitis induced by dextran sulfate sodium salt (DSS) and a Caco-2 cell model. The findings demonstrated that intervention with OPs effectively mitigated the symptoms associated with DSS-induced colitis. The physicochemical characterization analysis demonstrated that the molecular weight of the OPs was predominantly below 5 kDa, with a predominant composition of 266 peptides. This study provides further evidence of the regulatory impact of OPs on the Keap1-Nrf2 signaling axis and elucidates the potential role of WGVGVRAERDA as the primary bioactive peptide responsible for the functional effects of OPs. Ultimately, the results of this investigation demonstrate that OPs effectively mitigate DSS-induced colitis by preserving the integrity of the intestinal barrier and modulating the Keap1-Nrf2 axis. Consequently, these findings establish a theoretical foundation for the utilization of OPs as dietary supplements to prevent the onset of IBD.
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Colitis , Enfermedades Inflamatorias del Intestino , Humanos , Animales , Ratones , Avena , Sulfato de Dextran/efectos adversos , Factor 2 Relacionado con NF-E2/metabolismo , Células CACO-2 , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Colitis/inducido químicamente , Colitis/prevención & control , Colitis/metabolismo , Cloruro de Sodio/efectos adversos , Cloruro de Sodio Dietético/efectos adversos , Enfermedades Inflamatorias del Intestino/inducido químicamente , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , Colon/metabolismoRESUMEN
As the basis of animal reproductive activity, normal spermatogenesis directly determines the efficiency of livestock production. An in-depth understanding of spermatogenesis will greatly facilitate animal breeding efforts and male infertility treatment. With the continuous development and application of gene editing technologies, they have become valuable tools to study the mechanism of spermatogenesis. Gene editing technologies have provided us with a better understanding of the functions and potential mechanisms of action of factors that regulate spermatogenesis. This review summarizes the applications of gene editing technologies, especially CRISPR/Cas9, in deepening our understanding of the function of spermatogenesis-related genes and disease treatment. The problems of gene editing technologies in the field of spermatogenesis research are also discussed.
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Sistemas CRISPR-Cas , Infertilidad Masculina , Animales , Sistemas CRISPR-Cas/genética , Edición Génica , Humanos , Infertilidad Masculina/genética , Infertilidad Masculina/terapia , Masculino , Espermatogénesis/genética , TecnologíaRESUMEN
Mastitis, which affects milk quality and yield, is one of the most common diseases in dairy cows, causing large economic losses. Cow mastitis is classified into clinical and subclinical types. Subclinical mastitis presents without obvious lesions in the udder or noticeable change in milk samples, indicating persistent chronic infection that is difficult to detect and treat. Therefore, finding specific biomarkers is of great significance for the early diagnosis and treatment of subclinical mastitis. As mediators of intercellular communication, exosomes have been shown to be extensively involved in various physiological and pathological processes in the body. Exosomes in milk, blood, and cell supernatant can carry stable cell source-specific nucleic acids, proteins, and metabolites. Hence, exosomes show great application prospects for early diagnosis, targeted therapy, and disease mechanism analysis. In this review, we summarize the biogenesis, biological functions, and methods of isolating and identifying exosomes and review the current status of exosome research related to mastitis. Finally, in view of the application of exosomes to diagnose, treat, and perform disease mechanism analysis in mastitis, deficiencies in recent research on mastitis exosomes are described, and the direction of future exosome research efforts in mastitis is proposed.
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The incidence of kidney renal clear cell carcinoma (KIRC) is rising worldwide, and the prognosis is poor. Cuproptosis is a new form of cell death that is dependent on and regulated by copper ions. The relationship between cuproptosis and KIRC remains unclear. In the current study, changes in cuproptosis-related genes (CRGs) in TCGA-KIRC transcriptional datasets were characterized, and the expression patterns of these genes were analyzed. We identified three main molecular subtypes and discovered that multilayer CRG changes were associated with patient clinicopathological traits, prognosis, elesclomol sensitivity, and tumor microenvironment (TME) cell infiltration characteristics. Then, a CRG score was created to predict overall survival (OS). The CRG score was found to be strongly linked to the TME. These findings may help elucidate the roles of CRGs in KIRC, potentially enhancing understanding of cuproptosis and supporting the development of more effective immunotherapy strategies.
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Skin cutaneous melanoma (SKCM) is one of the most malignant and aggressive forms of cancer. Investigating the mechanisms of carcinogenesis further could lead to the discovery of prognostic biomarkers that could be used to guide cancer treatment. In this study, we conducted integrative bioinformatics analyses of TCGA database, STRING, cBioPortal, TRRUST, The Human Protein Atlas, and DGIdb to determine which hub genes contributed to tumor progression and the cancer-associated immunology of SKCM. The results show that immune-related 873 differential genes grouped SKCM samples into subtypes. The initial results showed that the optimal number of clusters was two subgroups. Further analysis showed that there were significant differences in survival rate and immune infiltration level between the two subgroups. Subsequently, obtaining the different genes between groups, construct PPI to screen 6 hub genes (HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DRA, HLA-DRB1, HLA-DRB5). In total, 6 MHC class II molecules were significantly related to overall survival. We then analyzed the expression of these genes along with their mutation landscapes, transcription factor regulation, and drug regulatory networks. In summary, our study identified 6 MHC class II molecules (HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DRA, HLA-DRB1, HLA-DRB5) as important biomarkers in the occurrence and progression of SKCM tumors. Their expression levels are closely related to prognosis and immune infiltration and can help us better understand the tumorigenesis of SKCM.
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The present study aimed to evaluate the antimicrobial activity of peppermint oil against Staphylococcus aureus, and further investigate the influence of peppermint oil on S. aureus virulence-related exoprotein production. The data show that peppermint oil, which contained high contents of menthone, isomenthone, neomenthol, menthol, and menthyl acetate, was active against S. aureus with minimal inhibitory concentrations (MICs) ranging from 64-256 µg/mL, and the production of S. aureus exotoxins was decreased by subinhibitory concentrations of peppermint oil in a dose-dependent manner. The findings suggest that peppermint oil may potentially be used to aid in the treatment of S. aureus infections.
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Exotoxinas/metabolismo , Aceites de Plantas/farmacología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/metabolismo , Animales , Antieméticos/farmacología , Antieméticos/uso terapéutico , Exotoxinas/genética , Cromatografía de Gases y Espectrometría de Masas/métodos , Hemólisis/efectos de los fármacos , Mentha piperita , Pruebas de Sensibilidad Microbiana , Aceites de Plantas/química , Aceites de Plantas/uso terapéutico , Conejos , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/patogenicidad , Transcripción Genética/efectos de los fármacosRESUMEN
OBJECTIVE: To investigate the effects of bone marrow mesenchymal stem cells transplantation on organoretinal cultures after a hypoxia injury. METHOD: The retinal tissues were cultured in vitro and then transplanted with bone marrow mesenchymal stem cells. Then, H&E staining and immunohistochemical assay were conducted to investigate the changes in retinal tissue structure and the migration and differentiation of stem cells. RESULTS: The retinal tissues were slightly damaged in the stem cell transplantation group; the control group, the retinal tissue structure was changed, and the thinning of their thickness was clearly evident. The transplanted stem cells can migrate to each layer of the retina to replace the damaged dead cells, which can protect the peripheral injured tissues and cells.
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Growth hormone (GH) is an important hormone released by the pituitary gland that plays a key role in the growth and development of organisms. In our study, TargetScan analysis and the dual luciferase reporter assays were used to predict and screen for miRNAs that might act on the rat Gh1 gene, and we identified miR-543-5p. Then, the GH3 cell line and the primary rat pituitary cells were transfected with miRNA mimic, inhibitor, and siRNA. We detected the Gh1 gene expression and the GH secretion by real-time PCR and ELISAs, respectively, to verify the regulatory effect of miR-543-5p on GH secretion. The results showed that miR-543-5p can inhibit Gh1 mRNA expression and reduce GH secretion. MiR-543-5p inhibitor upregulated Gh1 mRNA expression and increased GH secretion compared with the negative control. In summary, miR-543-5p downregulates Gh1 expression, resulting in a decrease in GH synthesis and secretion, which demonstrates the important role of miRNAs in regulating GH and animal growth and development.
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Hormona del Crecimiento/genética , MicroARNs/genética , Hormonas Adenohipofisarias/genética , Regiones no Traducidas 3'/genética , Animales , Línea Celular , Expresión Génica , Regulación de la Expresión Génica/genética , Hormona del Crecimiento/metabolismo , Masculino , Hipófisis/metabolismo , Adenohipófisis/metabolismo , Hormonas Adenohipofisarias/metabolismo , Cultivo Primario de Células , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , TransfecciónRESUMEN
Mice with spontaneous coat mutations are ideal animal models for studying skin development and tumorigenesis. In this study, skin hair growth cycle abnormalities were examined in NIH hairless mice 42 days after birth (P42) by using hematoxylin-eosin (H&E) staining. To examine the gene expression patterns in the skin of mutant mice, the dorsal skin of P42 female NIH mice and NIH hairless mice was sequenced by RNA-Seq, and 5,068 differentially expressed genes (DEGs) were identified (false discovery rate [FDR] ≥ 2, P < 0.05). A pathway analysis showed that basal cell carcinoma, the cell cycle and the Hippo, Hedgehog and Wnt signaling pathways were up-regulated in NIH hairless mice. Previous studies have shown that these pathways are closely associated with cell proliferation, cell cycle, organ size and cancer development. In contrast, signal transduction, bacterial and parasitic infection, and receptor-mediated pathways, including calcium signaling, were down-regulated in NIH hairless mice. A gene interaction network analysis was performed to identify genes related to hair follicle development. To verify the reliability of the RNA-Seq results, we used q-PCR to analyze 12 key genes identified from the gene interaction network analysis, including eight down-regulated and four up-regulated genes, and the results confirmed the reliability of the RNA-Seq results. Finally, we constructed the differential gene expression profiles of mutant mice by RNA-Seq. NIH hairless mice exhibited abnormalities in hair development and immune-related pathways. Pik3r1 and Pik3r3 were identified as key genes, laying the foundation for additional in-depth studies of hairless mice.
Asunto(s)
Perfilación de la Expresión Génica , Piel/metabolismo , Animales , Fosfatidilinositol 3-Quinasa Clase Ia/genética , Femenino , Ontología de Genes , Redes Reguladoras de Genes , Ratones , Ratones Pelados , Fenotipo , Análisis de Secuencia de ARNRESUMEN
Spontaneous mutant hairless (HL) mice are often used to study hair growth and hair follicle development, and they often exhibit immune dysfunctions. Listeria monocytogenes, an important food-borne bacterium, has been used in animal models to study immune responses to infection. Herein, we analyzed the innate immunity of HL mice and the impact of gut microbial polymorphisms on L. monocytogenes infection. Compared to NIH mice, NIH HL mice were more susceptible to L. monocytogenes, as weight losses, mortality, bacterial load, and histopathological lesions were more severe; the decrease in monocytes may be an important underlying reason. The degree of spleen damage was reduced after co-housing, indicating that the host guides the gut microbiota to alleviate infection. High-throughput pyrosequencing of 16S rRNA demonstrated that gut microbiota composition differed between NIH HL and NIH mice. Infection with L. monocytogenes induced an increase in the number of bacteria belonging to the Rikenellaceae family and Gammaproteobacteria class, and decreased bacteria belonging to the Clostridiales class and Lachnospiraceae family. A substantial reduction in Clostridiales bacteria in infected HL mice may cause a serious infection. The Mycoplasma genus was present only in NIH HL mice and was, thus, considered a biomarker. The results of this study improve our understanding of the use of NIH HL mice as a good animal model of innate immune dysfunction.