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Tuberculosis (TB) is still an urgent global public health problem. Notably, mucosal-associated invariant T (MAIT) cells play an important role in early anti-TB immune response. Targeted control of them may be an effective method to improve vaccine efficacy and TB treatment. However, the biology and signal regulation mechanisms of MAIT cells in TB patients are still poorly understood. Previous studies have been limited by the lack of reagents to specifically identify MAIT cells. In addition, the use of alternative markers may subsume non-MAIT cell into MAIT cell populations. In this study, the human MR1 tetramer which can specifically identify MAIT cells was used to further explore the effect and mechanism of MAIT cells in anti-TB immune response. Our results showed that the tetramer+ MAIT cells in peripheral blood of TB patients were mainly CD8+ or CD4-CD8- cells, and very few were CD4+ cells. After BCG infecting autologous antigen-presenting cells, MAIT cells in patients produced significantly higher levels of cytokines, lysis and proliferation compared with healthy controls. After suppression of mTORC1 by the mTORC1-specific inhibitor rapamycin, the immune response of MAIT cells in patients was significantly reduced. This study demonstrates that peripheral blood tetramer+ MAIT cells from TB patients have significant anti-TB immune effect, which is regulated by mTORC1. This could provide ideas and potential therapeutic targets for the development of novel anti-TB immunotherapy.
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Extensive research has been conducted on the role of CXCR3 in immune responses and inflammation. However, the role of CXCR3 in the reproductive system, particularly in oocyte development, remains unknown. In this study, we present findings on the involvement of CXCR3 in the meiotic division process of mouse oocytes. We found CXCR3 was expressed consistently throughout the entire maturation process of mouse oocyte. Inhibition of CXCR3 impaired the asymmetric division of oocyte, while the injection of Cxcr3 mRNA was capable of restoring these defects. Further study showed that inhibition of CXCR3 perturbed spindle migration by affecting LIMK/cofilin pathway-mediated actin remodeling. Knockout of CXCR3 led to an upregulation of actin-binding protein and an increased ATP level in GV-stage oocytes, while maintaining normal actin dynamics during the process of meiosis. Additionally, we noticed the expression level of DYNLT1 is markedly elevated in CXCR3-null oocytes. DYNLT1 bound with the Arp2/3 complex, and knockdown of DYNLT1 in CXCR3-null oocytes impaired the organization of cytoplasmic actin, suggesting the regulatory role of DYNLT1 in actin organization, and the compensatory expression of DYNLT1 may contribute to maintain normal actin dynamics in CXCR3-knockout oocytes. In summary, our findings provide insights into the intricate network of actin dynamics associated with CXCR3 during oocyte meiosis.
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Actinas , Oócitos , Receptores CXCR3 , Animais , Oócitos/metabolismo , Oócitos/fisiologia , Camundongos , Actinas/metabolismo , Actinas/genética , Receptores CXCR3/metabolismo , Receptores CXCR3/genética , Feminino , Meiose/fisiologia , Camundongos KnockoutRESUMO
To systematically evaluate the risk factors for wound infection at the surgical site after neurosurgical craniotomy by meta-analysis, and to provide an evidence-based basis for preventing the occurrence of wound infection. A computerised search of PubMed, EMBASE, Cochrane Library, China National Knowledge Infrastructure and Wanfang database was conducted for relevant studies on risk factors for surgical site wound infection after neurosurgical craniotomy published from the database inception to November 2023. Two researchers independently screened the literature, extracted the data and performed quality assessment in strict accordance with the inclusion and exclusion criteria. STATA 17.0 software was applied for data analysis. Overall, 18 papers with 17 608 craniotomy patients were included, of which 905 patients developed wound infections. The analysis showed that underlying diseases [OR = 2.50, 95% CI (1.68, 3.72), p < 0.001] and emergency surgery [OR = 2.47, 95% CI (1.80, 3.38), p < 0.001] were the risk factors for developing wound infections after craniotomy, age < 60 years [OR = 0.72, 95% CI (0.52, 0.98), p = 0.039] was a protective factor for wound infections; whereas sex [OR = 1.11, 95% CI (0.98, 1.27), p = 0.112] and the antimicrobial use [OR = 1.30, 95% CI (0.81 2.09), p = 0.276] were not associated with the presence or absence of wound infection after craniotomy. Underlying disease and emergency surgery are risk factors for developing wound infections after craniotomy, whereas age < 60 years is a protective factor. Clinicians can reduce the occurrence of postoperative wound infections by communicating with patients in advance about the possibility of postoperative wound infections based on these factors, and by doing a good job of preventing postoperative wound infections.
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Craniotomia , Procedimentos Neurocirúrgicos , Infecção da Ferida Cirúrgica , Humanos , Infecção da Ferida Cirúrgica/prevenção & controle , Infecção da Ferida Cirúrgica/etiologia , Infecção da Ferida Cirúrgica/epidemiologia , Fatores de Risco , Procedimentos Neurocirúrgicos/efeitos adversos , Craniotomia/efeitos adversos , Feminino , Masculino , Fatores de Proteção , Pessoa de Meia-Idade , Adulto , IdosoRESUMO
BACKGROUND: Chimeric antigen receptor CAR-T cell therapies have ushered in a new era of treatment for specific blood cancers, offering unparalleled efficacy in cases of treatment resistance or relapse. However, the emergence of cytokine release syndrome (CRS) as a side effect poses a challenge to the widespread application of CAR-T cell therapies. Melatonin, a natural hormone produced by the pineal gland known for its antioxidant and anti-inflammatory properties, has been explored for its potential immunomodulatory effects. Despite this, its specific role in mitigating CAR-T cell-induced CRS remains poorly understood. METHODS: In this study, our aim was to investigate the potential of melatonin as an immunomodulatory agent in the context of CD19-targeting CAR-T cell therapy and its impact on associated side effects. Using a mouse model, we evaluated the effects of melatonin on CAR-T cell-induced CRS and overall survival. Additionally, we assessed whether melatonin administration had any detrimental effects on the antitumor efficacy and persistence of CD19 CAR-T cells. RESULTS: Our findings demonstrate that melatonin effectively mitigated the severity of CAR-T cell-induced CRS in the mouse model, leading to improved overall survival outcomes. Remarkably, melatonin administration did not compromise the antitumor effectiveness or persistence of CD19 CAR-T cells, indicating its compatibility with therapeutic goals. These results suggest melatonin's potential as an immunomodulatory compound to alleviate CRS without compromising the therapeutic benefits of CAR-T cell therapy. CONCLUSION: The study's outcomes shed light on melatonin's promise as a valuable addition to the existing treatment protocols for CAR-T cell therapies. By attenuating CAR-T cell-induced CRS while preserving the therapeutic impact of CAR-T cells, melatonin offers a potential strategy for optimizing and refining the safety and efficacy profile of CAR-T cell therapy. This research contributes to the evolving understanding of how to harness immunomodulatory agents to enhance the clinical application of innovative cancer treatments.
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Síndrome da Liberação de Citocina , Imunoterapia Adotiva , Melatonina , Antígenos CD19 , Terapia Baseada em Transplante de Células e Tecidos , Síndrome da Liberação de Citocina/terapia , Fatores Imunológicos/farmacologia , Imunoterapia Adotiva/efeitos adversos , Melatonina/farmacologia , Recidiva Local de Neoplasia , Receptores de Antígenos de Linfócitos T , Receptores de Antígenos Quiméricos , Animais , CamundongosRESUMO
3-Nitropropionic acid (3-NP) is a mycotoxin commonly found in plants and fungi that has been linked to mammalian intoxication. Previously, we found 3-NP treatment exhibited reproductive toxicity by inducing oxidative stress in mouse ovary; however, the toxic effects of 3-NP on mouse oocyte maturation have not been investigated. Sulforaphane (SFN) is a naturally bioactive phytocompound derived from cruciferous vegetables that has been shown to possess cytoprotective properties. The present study was designed to investigate the cytotoxicity of 3-NP during mouse oocyte maturation and the protective effects of SFN on oocytes challenged with 3-NP. The results showed 3-NP had a dose-dependent inhibitory effect on oocyte maturation, and SFN significantly alleviated the defects caused by 3-NP, including failed first polar body extrusion and abnormal spindle assembly. Furthermore, 3-NP caused abnormal mitochondrial distribution in oocytes and disrupted mitochondrial functions, including mitochondrial depolarization, decreased ATP levels, and increased mitochondrial-derived ROS. Finally, 3-NP induced oxidative stress in oocytes, leading to increased apoptosis and autophagy, while SFN supplementation had significant cytoprotective effects on these damages. Collectively, our results provide insight on the mechanism of 3-NP toxicity in mouse oocytes and suggest the application of SFN may be a viable intervention strategy to mitigate 3-NP-induced reproductive toxicity.
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Oócitos , Estresse Oxidativo , Feminino , Animais , Camundongos , Meiose , Apoptose , MamíferosRESUMO
BACKGROUND: The Makorin ring finger protein 1 (MKRN1) gene, also called RNF61, is located on the long arm of chromosome 7 and is a member of the RING finger protein family. The E3 ubiquitin ligase MKRN1 is closely linked to tumour development, but the exact mechanism needs to be elucidated. In this study, we aimed to investigate the specific mechanism and role of MKRN1 in colorectal cancer (CRC) development. METHODS: MKRN1 expression in CRC was analysed using the Cancer Cell Line Encyclopaedia and the Cancer Genome Atlas (TCGA) databases. Rectal tumour tissues were frozen to explore the MKRN1 expression in CRC and its clinical significance. The impact of MKRN1 on CRC cell proliferation and migration was observed using CCK8, colony formation, wound healing, and transwell assays. A combination of MKRN1 quantitative proteomics, ubiquitination modification omics analysis, and a string of in vitro and in vivo experiments revealed the potential mechanisms by which MKRN1 regulates CRC metastasis. RESULTS: MKRN1 expression was significantly elevated in CRC tissues compared to paracancerous tissues and was positively linked with prognosis (P < 0.01). MKRN1 downregulation inhibits CRC cell proliferation, migration, and invasion. Conversely, MKRN1 overexpression promotes the proliferation, migration, and invasion of CRC cells. Mechanistically, MKRN1 induces epithelial-mesenchymal transition (EMT) in CRC cells via ubiquitination and degradation of Smad nuclear-interacting protein 1 (SNIP1). Furthermore, SNIP1 inhibits transforming growth factor-ß (TGF-ß) signalling, and MKRN1 promotes TGF-ß signalling by degrading SNIP1 to induce EMT in CRC cells. Finally, using conditional knockout mice, intestinal lesions and metastatic liver microlesions were greatly reduced in the intestinal knockout MKRN1 group compared to that in the control group. CONCLUSIONS: High MKRN1 levels promote TGF-ß signalling through ubiquitination and degradation of SNIP1, thereby facilitating CRC metastasis, and supporting MKRN1 as a CRC pro-cancer factor. The MKRN1/SNIP1/TGF-ß axis may be a potential therapeutic target in CRC.
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Neoplasias Colorretais , Proteínas de Ligação a RNA , Ribonucleoproteínas , Animais , Camundongos , Linhagem Celular , Proliferação de Células , Neoplasias Colorretais/genética , Proteólise , Humanos , Ribonucleoproteínas/metabolismo , Proteínas de Ligação a RNA/genética , Fator de Crescimento Transformador beta/metabolismo , Transdução de SinaisRESUMO
To verify the inhibitory mechanism of ß-catenin-designed peptides in colorectal cancer(CRC) tumors, the following experiments were performed. In vitro colony formation, Transwell assays, and flow cytometry were performed to assess the biological effects of designed peptides (F18KD, F20A4-7k, F20A4-10k, and F20A3-9k + F20A4-10k + F20A5-9k) in HT-29 cells. In vivo xenograft experiments were performed and treated with peptides. Next, tumors were subjected to Hematoxylin and eosin staining (HE), immunohistochemical, and terminal deoxynucleotidyl transferase dUTP nick end labeling staining assays to evaluate the inhibitory effect of peptides on tumors. ß-Catenin levels were quantified via western blotting (WB) and quantitative real-time polymerase chain reaction, and ß-catenin was located using confocal laser scanning microscopy. T-cell factor-4 (TCF-4), C-myc, and CCND1 levels were quantified via WB. Results were obtained as following. First, the peptides reduced viability, migration, and invasion; promoted apoptosis; and stabilized the S phase of HT-29 cells. Second, peptides suppressed tumor growth and downregulated the expression of CD34, vascular endothelial growth factor, and ß-catenin in tumors. Furthermore, we found that peptides downregulated ß-catenin expression in both the cytoplasm and nucleus; TCF-4, C-myc, and CCND1 expression was also downregulated. Notably, ß-catenin-targeting peptides had a better inhibitory effect on CRC than non-ß-catenin-target peptides, and a combination of peptides exerted a more potent inhibitory effect on CRC than single peptides. It suggested that ß-Catenin-targeting peptides promote apoptosis in CRC tumors by inhibiting activation of the Wnt/ß-catenin pathway.
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Neoplasias Colorretais , Fator A de Crescimento do Endotélio Vascular , Humanos , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/genética , Via de Sinalização Wnt , Apoptose , Peptídeos/farmacologia , Peptídeos/metabolismo , Proliferação de Células , Linhagem Celular Tumoral , Movimento Celular , Regulação Neoplásica da Expressão GênicaRESUMO
Harmful Microcystis blooms (HMBs) and microcystins (MCs) that are produced by Microcystis seriously threaten water ecosystems and human health. This study demonstrates an eco-friendly strategy for simultaneous removal of MCs and HMBs by adopting unique hyperoxic graphene oxides (HGOs) as carrier and pure microcystinase A (PMlrA) as connecting bridge to form stable HGOs@MlrA composite. After oxidation, HGOs yield inherent structural strain effects for boosting the immobilization of MlrA by material characterization and density functional theory calculations. HGO5 exhibits higher loading capacities for crude MlrA (1,559 mg·g-1) and pure MlrA (1,659 mg·g-1). Moreover, the performances of HGO5@MlrA composite, including the capability of removing MCs and HMBs, the ecological and human safety compared to MlrA or HGO5 treatment alone, have been studied. These results indicate that HGO5 can be used as a promising candidate material to effectively improve the application potential of MlrA in the simultaneous removal of MCs and HMBs.
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Background: Among central nervous system tumors, glioblastoma (GBM) is considered to be the most destructive malignancy. Recurrence is one of the most fatal aspects of GBM. However, the driver molecules that trigger GBM recurrence are currently unclear. Methods: The mRNA expression data and clinical information of GBM and normal tissues were collected from the Chinese Glioma Genome Atlas The Cancer Genome Atlas (TCGA), and REpository for Molecular BRAin Neoplasia DaTa (REMBRANDT) cohorts. The DESeq2 R package was used to identify the differentially expressed genes between primary and recurrent GBM. ClueGO, Kyoto Encyclopedia of Genes and Genomes (KEGG), Biological Process in Gene ontology (GO-BP), and the Protein ANalysis THrough Evolutionary Relationships (PANTHER) pathway analyses were performed to explore the enriched signaling pathways in upregulated DEGs in recurrent GBM. A gene list that contained potential oncogenes that showed a significant negative correlation with patient survival from The Cancer Genome Atlas was used to further screen driver candidates for recurrent GBM. Univariate Cox proportional hazards regression analyses were used to investigate the risk score for the mRNA expression of the candidates. Single-cell RNA sequencing (scRNA-Seq) analyses were used to determine the cell type-specific distribution of Fc gamma receptor II b (FcγRIIb) in GBM. Immunohistochemistry (IHC) was used to confirm the FcγRIIb-positive cell populations in primary and paired recurrent GBM. Results: Through DEG analysis and overlap analysis, a total of 10 genes that are upregulated in recurrent GBM were screened. Using validation databases, FcγRIIb was identified from the 10 candidates that may serve as a driver for recurrent GBM. FCGR2B expression, not mutation, further showed a highly negative correlation with the poor prognosis of patients with recurrent GBM. Furthermore, scRNA-Seq analyses revealed that tumor-associated macrophage- and dendritic cell-specific FCGR2B was expressed. Moreover, FcγRIIb also showed a strong positive correlation coefficient with major immune-associated signaling pathways. In clinical specimens, FcγRIIb-positive cell populations were higher in recurrent GBM than in primary GBM. Conclusion: This study provides novel insights into the role of FcγRIIb in recurrent GBM and a promising strategy for treatment as an immune therapeutic target.
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Chronic stress affects the reproductive health of mammals; however, the impact of adrenocorticotropin hormone (ACTH) level elevation during chronic stress on the reproduction of weaned sows remains unclear. In this study, nine weaned sows with the same parturition date were randomly divided into control group (n = 4) and ACTH group (n = 5). Each group received intravenous administration of ACTH three times daily for 7 days. Blood samples were collected every 3 h after injection. A radioimmunoassay was used to measure the concentrations of cortisol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P4) and estradiol-17ß (E2) in the blood. Estrus was determined according to changes in the vulva and the boar contact test. The mRNA expressions of glucocorticoid receptor, FSH receptor, LH receptor (LHR) in the corpus luteum (CL) were detected by qRT-PCR. The results showed that ACTH administration substantially delayed the initiation of estrus and the pre-ovulatory LH peak. The sows of control group ovulated within 10 days and the ovulation rate was 100%, while it was 60% in the ACTH group. Two sows of ACTH group showed pseudo-estrus. The E2 concentrations significantly decreased in the ACTH group at 36 h, 42 h and 66 h of the experimental period. The P4 concentrations in the ACTH group significantly decreased at 132, 138, and 147 h of the experimental period. ACTH significantly reduced the LHR mRNA expression in CLs. In conclusion, long-term repeated ACTH administration affects the endocrinology, estrus onset, and ovarian function of weaned sows.
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Hormônio Adrenocorticotrópico , Estro , Hormônio Adrenocorticotrópico/farmacologia , Animais , Estradiol , Estro/fisiologia , Feminino , Hormônio Luteinizante , Mamíferos/metabolismo , Ovulação , Progesterona , Suínos , DesmameRESUMO
In mammalian ovaries, the avascular environment within follicular cavity is supposed to cause hypoxic status in granulosa cells (GCs), leading to apoptotic cell death accompanied by cumulative reactive oxygen species (ROS) production. Melatonin (N-acetyl-5-methoxytryptamine, MT), a broad-spectrum antioxidant that exists in porcine follicle fluid, was suggested to maintain GCs survival under stress conditions. In this study, using the established hypoxic model (1% O2) of cultured porcine GCs, we explored the effect of MT on GCs apoptosis. The results showed that MT restored cell viability and reduced the apoptosis of GCs during hypoxia exposure. In addition, GCs treated with MT exhibited decreased ROS levels and increased expression of antioxidant enzymes including heme oxygenase-1 (HO-1), glutathione S-transferase (GST), superoxide dismutase 1 (SOD1), and catalase (CAT) upon hypoxia incubation. Moreover, the hypoxia-induced expression of cleaved caspase 3, 8, and 9 was significantly inhibited after MT treatment. In contrast, blocking melatonin receptor 2 (MTNR1B) with a competitive antagonist 4-phenyl-2-propionamidotetralin (4P-PDOT) diminished the inhibitory effects of MT on caspase 3 activation. By detecting levels of protein kinase (PKA), a downstream kinase of MTNR1B, we further confirmed the involvement of MT-MTNR1B signaling in mediating GCs protection during hypoxia stress. Together, the present data provide mechanistic evidence suggesting the role of MT in defending GCs from hypoxia-induced apoptosis.
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The formation of zygote is the beginning of mammalian life, and dynamic epigenetic modifications are essential for mammalian normal development. H3K27 di-methylation (H3K27me2) and H3K27 tri-methylation (H3K27me3) are marks of facultative heterochromatin which maintains transcriptional repression established during early development in many eukaryotes. However, the mechanism underlying establishment and regulation of epigenetic asymmetry in the zygote remains obscure. Here we show that maternal EZH2 is required for the establishment of H3K27me3 in mouse zygotes. However, combined immunostaining with ULI-NChIP-seq (ultra-low-input micrococcal nuclease-based native ChIP-seq) shows that EZH1 could partially safeguard the role of EZH2 in the formation of H3K27me2. Meanwhile, we identify that EHMT1 is involved in the establishment of H3K27me2, and that H3K27me2 might be an essential prerequisite for the following de novo H3K27me3 modification on the male pronucleus. In this work, we clarify the establishment and regulatory mechanisms of H3K27me2 and H3K27me3 in mouse zygotes.
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Genoma , Histona-Lisina N-Metiltransferase/metabolismo , Histonas/metabolismo , Zigoto/metabolismo , Animais , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Epigenômica , Heterocromatina , Histona-Lisina N-Metiltransferase/genética , Masculino , Metilação , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout , Nuclease do Micrococo , Oogênese/fisiologia , Complexo Repressor Polycomb 2/genética , Processamento de Proteína Pós-TraducionalRESUMO
Octamer-binding transcription factor 4 (Oct4) has been recently implicated as a proangiogenic regulator in several induced pluripotent stem cells (iPSCs), however, its role in cancer stem-like cells (CSCs) remain unclear. We report here that Oct4 participates in tumor vasculogenesis in liver CSCs (LCSCs). We identify that LCSCs possess the potential of endothelial trans-differentiation under endothelial induction, present endothelial specific markers and their functions in vitro, and participate in neovasculogenesis in vivo. The knockdown of the Oct4A by short hairpin RNA (shRNA) in LCSCs represses endothelial trans-differentiation potential, but induces endothelial lineage-restricted differentiation, the latter is positively regulated by Oct4B1. Furthermore, Oct4 regulates vasculogenesis in LCSCs may be via the AKT-NF-κB-p65 signaling pathway. This work reveals Oct4, which is a crucial regulator, plays a critical role in tumor endothelial-like cells transition of LCSCs through Oct4A and Oct4B1 by different ways. The simultaneous inhibition of both the isoforms of Oct4 is hence expected to help regress neovascularization derived from CSCs. Our findings may provide insights to the possible new mechanisms of tumor vasculogenesis for primary liver cancer.
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Mineral elements and stable isotopes combined with stoichiometric methods were used as a potential tool for first authenticating Chinese tea according to it's production year. A total of 86 mineral elements and stable isotope compositions were determined from the Xiangzhujing Pu'er tea in five different production years using ICP-MS and ICP-OES. On the basis of 78 statistically significant mineral elements and stable isotopes, HCA, PCA, PLS-DA, BP-ANN, and LDA were employed to build authentication models for predicting the Pu'er tea with different production years. The clustering results of the HCA and PCA were worse than that of PLS-DA, BP-ANN, and LDA. The PLS-DA model displayed a perfect model performance (R2X = 0.86, R2Y = 0.974, and Q2 = 0.922). The authentication performance of LDA and BP-ANN revealed their 100% recognition sensitivity and prediction ability and was thus better than that of PLS-DA. Mn, 68Zn, and 203Tl were the markers for enabling the successful authentication of Pu'er tea with different production years. This study contributes toward generalizing the use of mineral element and stable isotope fingerprinting combined with LDA and BP-ANN as a promising tool for authentication of tea worldwide.
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Camellia sinensis , Chá , Análise por Conglomerados , Isótopos , Análise EspectralRESUMO
BACKGROUND: There is an urgent need to strengthen the testing and certification of geographically iconic foods, as well as to use discriminatory science and technology for their regulation and verification. Multi-element and stable isotope analyses were combined to provide a new chemometric approach for improving the discrimination tea samples from different geographical origins. Different stoichiometric methods [principal component analysis (PCA), hierarchical cluster analysis (HCA), partial least squares-discriminant analysis (PLS-DA), back propagation based artificial neural network (BP-ANN) and linear discriminant analysis (LDA)] were used to demonstrate this discrimination approach using Yongchuanxiuya tea samples in an experimental test. RESULTS: Multi-element and stable isotope analyses of tea samples using inductively coupled plasma mass spectrometry and inductively coupled plasma optical emission spectrometry easily distinguished the geographical origins. However, the clustering ability of the two unsupervised learning methods (PCA and HCA) were worse compared to that of the three supervised learning methods (PLS-DA, BP-ANN and LDA). BP-ANN and LDA, with 100% recognition and prediction abilities, were found to be better than PLS-DA. 86 Sr and 112 Cd were the markers enabling the successful classification of tea samples according to their geographical origins. Under the validation by 'blind' dataset, the prediction accuracies of the BP-ANN and LDA methods were all greater than 90%. The LDA method showed the best performance, with an accuracy of 100%. CONCLUSION: In summary, determination of mineral elements and stable isotopes using inductively coupled plasma mass spectrometry and inductively coupled plasma optical emission spectrometry techniques coupled with chemometric methods, especially the LDA method, is a good approach for improving the authentication of a diverse range of tea. The present study contributes toward generalizing the use of fingerprinting mineral elements and stable isotopes as a promising tool for testing the geographic roots of tea and food worldwide. © 2020 Society of Chemical Industry.
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Camellia sinensis/química , Espectrometria de Massas/métodos , Análise Espectral/métodos , Chá/química , Análise Discriminante , Geografia , Isótopos/química , Folhas de Planta/química , Análise de Componente Principal , Oligoelementos/químicaRESUMO
The ATP binding cassette (ABC) E1 (ABCE1), a member of the ABC family, was originally described as the RNase L inhibitor. Through forming a heterodimer with RNase L, ABCE1 participates in the negative regulation of the 2-5A/RNase L system and thus mediates a wide range of biological functions. Recent evidence has shown the new roles of ABCE1 in tumorigenesis. However, there have been no investigations on the specific effect of ABCE1 on glioma. In the present study, we examined the expression pattern and possible role of ABCE1 in glioma. Our study demonstrated that ABCE1 was up-regulated in glioma tissues and cell lines. Down-regulation of ABCE1 inhibited temozolomide (TMZ) resistance of glioma cells in vitro and in vivo In addition, we found that the PI3K/Akt/NF-κB pathway was involved in ABCE1-mediated chemoresistance of glioma cells. Taken together, our study suggested ABCE1 as a promising target for glioma chemotherapy.
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Transportadores de Cassetes de Ligação de ATP/genética , Antineoplásicos Alquilantes/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Regulação para Baixo , Resistencia a Medicamentos Antineoplásicos , Glioma/tratamento farmacológico , Temozolomida/farmacologia , Animais , Antineoplásicos Alquilantes/uso terapêutico , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Linhagem Celular Tumoral , Glioma/genética , Glioma/metabolismo , Humanos , Masculino , Camundongos Nus , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Temozolomida/uso terapêuticoRESUMO
BACKGROUND: The study aimed to evaluate the efficacy and safety of gingival mesenchymal stem cells (GMSCs) from human fetal gingival tissue used for treating gingival defects in a rat model. METHODS: GMSCs were isolated from human fetal gingival tissue and identified by flow cytometry for nestin, Oct4, vimentin, NANOG, CD105, and CD90. The immunogenicity of GMSCs was analyzed by mixed lymphocyte reactions; the tumorigenicity of GMSCs was evaluated by xenotransplanting into nude mice. The gingival defect animal model was established by mechanical resection in rats. GMSCs were transplanted into the defective area, and the regeneration of gingival tissue was observed twice weekly. Four weeks after transplantation, the gingival tissue was surgically cut down, and the graft was analyzed by immunohistochemistry staining for human mitochondrial antigens and rat CD3 and CD20. RESULTS: GMSCs from human fetal gingival tissue positively expressed nestin, Oct4, vimentin, NANOG, CD105, and CD90. There was no cell aggregation after mixed lymphocyte reactions, and interleukin-2 did not increase. Inoculation of GMSCs into nude mice for 6 months showed no tumor formation. GMSCs were transplanted into the gingiva defects of rats. One week after transplantation, the defect area was reduced, and after 3 weeks the morphology and color of local gingival tissue was similar to normal gingival tissue, and gingival height was the same as the normal control group. CONCLUSIONS: Using GMSCs from human fetal gingival tissue to treat gingival defects is a safe and effective innovative treatment method.
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Antígenos de Diferenciação/biossíntese , Feto , Gengiva , Doenças da Gengiva , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Animais , Gengiva/lesões , Gengiva/metabolismo , Gengiva/patologia , Doenças da Gengiva/metabolismo , Doenças da Gengiva/patologia , Doenças da Gengiva/terapia , Xenoenxertos , Humanos , Masculino , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Ratos , Ratos WistarRESUMO
LIM kinases (LIMK1/2) are LIM domain-containing serine/threonine/tyrosine kinases that mediate multiple cellular processes in mitosis. In the present study, we explored the functional roles and potential signaling pathway of LIMK1/2 during mouse oocyte meiosis. Disruption of LIMK1/2 activity and expression significantly decreased oocyte polar body extrusion. Live-cell imaging revealed that spindle migration was disturbed after both LIMK1 and LIMK2 knock down, and this might be due to aberrant distribution of actin filaments in the oocyte cytoplasm and cortex. Meanwhile, our results demonstrated that the function of LIMK1 and LIMK2 in actin assembly was related to cofilin phosphorylation levels. In addition, disruption of LIMK1/2 activity significantly increased the percentage of oocytes with abnormal spindle morphologies, which was confirmed by the abnormal p-MAPK localization. We further, explored the upstream molecules of LIMK1/2, and we found that after depletion of ROCK, phosphorylation of LIMK1/2 and cofilin were significantly decreased. Moreover, RhoA inhibition caused the decreased expression of ROCK, p-LIMK1/2, and cofilin. In summary, our results indicated that the small GTPase RhoA regulated LIMK1/2-cofilin to modulate cytoskeletal dynamics during mouse oocyte meiosis.
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Fatores de Despolimerização de Actina/metabolismo , Quinases Lim/metabolismo , Meiose/fisiologia , Oócitos/metabolismo , Quinases Associadas a rho/metabolismo , Actinas/metabolismo , Animais , Citoplasma/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Proteínas dos Microfilamentos/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Mitose/fisiologia , Fosforilação/fisiologia , Corpos Polares/metabolismo , Transdução de Sinais/fisiologia , Fuso Acromático/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismoRESUMO
Warburg effect, characterized by enhanced glycolysis and lactate production, even under aerobic conditions, is one of the hallmarks of cancer cells. However, the mechanism underlying this phenomenon remains poorly understood. Previous studies have shown that microRNA-150 (miR-150) is significantly up-regulated in various malignancies and represents a putative onco-miRNA in human cancers. In the present study, we aim to investigate the functional significance and molecular target of miR-150 in glioma. As a result, von Hippel-Lindau (VHL), which is a specific E3 ligase for hypoxia inducible factor 1 (HIF1α), was identified as a novel target of miR-150. Consistently, cells overexpressing miR-150 exhibited a metabolic shift, including enhanced glucose uptake and lactate production, which led to a rapid growth of glioma cells. Therefore, our results suggest that miR-150 modulates the Warburg effect in glioma via VHL/HIF1α and might provide a novel option for future treatments for glioma.
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OBJECTIVE: Recent evidence has suggested that circulating endothelial progenitor cells (EPCs) can repair the arterial endothelium during vascular injury. However, a reliable source of human EPCs is needed for therapeutic applications. In this study, we isolated human fetal aorta (HFA)-derived EPCs and analyzed the capacity of EPCs to differentiate into endothelial cells. In addition, because microvascular dysfunction is considered to be the major cause of diabetic foot (DF), we investigated whether transplantation of HFA-derived EPCs could treat DF in a rat model. METHODS: EPCs were isolated from clinically aborted fetal aorta. RT-PCR, fluorescence-activated cell sorting, immunofluorescence, and an enzyme-linked immunosorbent assay were used to examine the expressions of CD133, CD34, CD31, Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), von Willebrand Factor (vWF), and Endothelial Leukocyte Adhesion Molecule-1 (ELAM-1). Morphology and Dil-uptake were used to assess function of the EPCs. We then established a DF model by injecting microcarriers into the hind-limb arteries of Goto-Kakizaki rats and then transplanting the cultured EPCs into the ischemic hind limbs. Thermal infrared imaging, oxygen saturation apparatus, and laser Doppler perfusion imaging were used to monitor the progression of the disease. Immunohistochemistry was performed to examine the microvascular tissue formed by HFA-derived EPCs. RESULTS: We found that CD133, CD34, and VEGFR2 were expressed by HFA-derived EPCs. After VEGF induction, CD133 expression was significantly decreased, but expression levels of vWF and ELAM-1 were markedly increased. Furthermore, tube formation and Dil-uptake were improved after VEGF induction. These observations suggest that EPCs could differentiate into endothelial cells. In the DF model, temperature, blood flow, and oxygen saturation were reduced but recovered to a nearly normal level following injection of the EPCs in the hind limb. Ischemic symptoms also improved. Injected EPCs were preferentially and durably engrafted into the blood vessels. In addition, anti-human CD31+-AMA+-vWF+ microvasculars were detected after transplantation of EPCs. CONCLUSION: Early fetal aorta-derived EPCs possess strong self-renewal ability and can differentiate into endothelial cells. We demonstrated for the first time that transplanting HFA-derived EPCs could ameliorate DF prognosis in a rat model. These findings suggest that the transplantation of HFA-derived EPCs could serve as an innovative therapeutic strategy for managing DF.