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Zinc and RING finger 3 (ZNRF3) is a negative-feedback regulator of Wnt/ß-catenin signaling, which plays an important role in human brain development. Although somatically frequently mutated in cancer, germline variants in ZNRF3 have not been established as causative for neurodevelopmental disorders (NDDs). We identified 12 individuals with ZNRF3 variants and various phenotypes via GeneMatcher/Decipher and evaluated genotype-phenotype correlation. We performed structural modeling and representative deleterious and control variants were assessed using in vitro transcriptional reporter assays with and without Wnt-ligand Wnt3a and/or Wnt-potentiator R-spondin (RSPO). Eight individuals harbored de novo missense variants and presented with NDD. We found missense variants associated with macrocephalic NDD to cluster in the RING ligase domain. Structural modeling predicted disruption of the ubiquitin ligase function likely compromising Wnt receptor turnover. Accordingly, the functional assays showed enhanced Wnt/ß-catenin signaling for these variants in a dominant negative manner. Contrarily, an individual with microcephalic NDD harbored a missense variant in the RSPO-binding domain predicted to disrupt binding affinity to RSPO and showed attenuated Wnt/ß-catenin signaling in the same assays. Additionally, four individuals harbored de novo truncating or de novo or inherited large in-frame deletion variants with non-NDD phenotypes, including heart, adrenal, or nephrotic problems. In contrast to NDD-associated missense variants, the effects on Wnt/ß-catenin signaling were comparable between the truncating variant and the empty vector and between benign variants and the wild type. In summary, we provide evidence for mirror brain size phenotypes caused by distinct pathomechanisms in Wnt/ß-catenin signaling through protein domain-specific deleterious ZNRF3 germline missense variants.
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BACKGROUND: Reports of dual carriers of pathogenic BRCA1 variants in trans are extremely rare, and so far, most individuals have been associated with a Fanconi Anemia-like phenotype. METHODS: We identified two families with a BRCA1 in-frame exon 20 duplication (Ex20dup). In one male individual, the variant was in trans with the BRCA1 frameshift variant c.2475delC p.(Asp825Glufs*21). We performed splicing analysis and used a transcription activation domain (TAD) assay to assess the functional impact of Ex20dup. We collected pedigrees and mapped the breakpoints of the duplication by long- and short-read genome sequencing. In addition, we performed a mitomycin C (MMC) assay from the dual carrier using cultured lymphoblastoid cells. RESULTS: Genome sequencing and RNA analysis revealed the BRCA1 exon 20 duplication to be in tandem. The duplication was expressed without skipping any one of the two exon 20 copies, resulting in a lack of wild-type transcripts from this allele. TAD assay indicated that the Ex20dup variant has a functional level similar to the well-known moderate penetrant pathogenic BRCA1 variant c.5096G > A p.(Arg1699Gln). MMC assay of the dual carrier indicated a slightly impaired chromosomal repair ability. CONCLUSIONS: This is the first reported case where two BRCA1 variants with demonstrated functional impact are identified in trans in a male patient with an apparently normal clinical phenotype and no BRCA1-associated cancer. The results pinpoint a minimum necessary BRCA1 protein activity to avoid a Fanconi Anemia-like phenotype in compound heterozygous status and yet still predispose carriers to hormone-related cancers. These findings urge caution when counseling families regarding potential Fanconi Anemia risk. Furthermore, prudence should be taken when classifying individual variants as benign based on co-occurrence in trans with well-established pathogenic variants.
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Neoplasias da Mama , Anemia de Fanconi , Humanos , Masculino , Proteína BRCA1/genética , Éxons/genética , Anemia de Fanconi/genética , Mitomicina , FenótipoRESUMO
PURPOSE: Achieving a pathologic complete response (pCR) after neoadjuvant chemoradiotherapy (NCRT) remains a challenge for most patients with rectal cancer. Exploring the potential of combining NCRT with immunotherapy or targeted therapy for those achieving a partial response (PR) offers a promising avenue to enhance treatment efficacy. This study investigated the impact of NCRT on the tumor microenvironment in locally advanced rectal cancer (LARC) patients who exhibited a PR. METHODS: This was a retrospective, observational study. Five patients demonstrating a PR after neoadjuvant treatment for LARC were enrolled in the study. Biopsy samples before treatment and resected specimens after treatment were stained with a panel of 26 antibodies targeting various immune and tumor-related markers, each labeled with distinct metal tags. The labeled samples were then analyzed using the Hyperion imaging system. RESULTS: Heterogeneity within the tumor microenvironment was observed both before and after NCRT. Notably, tumor-associated macrophages, CD4 + T cells, CD8 + T cells, CD56 + natural killer cells, tumor-associated neutrophils, cytokeratin, and E-cadherin exhibited slight increase in abundance within the tumor microenvironment following treatment (change ratios = 0.78, 0.2, 0.27, 0.32, 0.17, 0.46, 0.32, respectively). Conversely, the number of CD14 + monocytes, CD19 + B cells, CD45 + CD4 + T cells, collagen I, α-smooth muscle actin, vimentin, and ß-catenin proteins displayed significant decreases post-treatment (change ratios = 1.73, 1.92, 1.52, 1.25, 1.52, 1.12, 2.66, respectively). Meanwhile, Foxp3 + regulatory cells demonstrated no significant change (change ratio = 0.001). CONCLUSIONS: NCRT has diverse effects on various components of the tumor microenvironment in LARC patients who achieve a PR after treatment. Leveraging combination therapies may optimize treatment outcomes in this patient population.
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Terapia Neoadjuvante , Neoplasias Retais , Microambiente Tumoral , Humanos , Neoplasias Retais/terapia , Neoplasias Retais/patologia , Neoplasias Retais/tratamento farmacológico , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Quimiorradioterapia , Resultado do Tratamento , Estudos RetrospectivosRESUMO
We have recently reported that transcription factor Runx3 is required for pulmonary generation of CD8+ cytotoxic T lymphocytes (CTLs) that play a crucial role in the clearance of influenza A virus (IAV). To understand the underlying mechanisms, we determined the effects of Runx3 knockout (KO) on CD8+ T cell local expansion and phenotypes using an inducible general Runx3 KO mouse model. We found that in contrast to the lungs, Runx3 general KO promoted enlargement of lung-draining mediastinal lymph node (mLN) and enhanced CD8+ and CD4+ T cell expansion during H1N1 IAV infection. We further found that Runx3 deficiency greatly inhibited core 2 O-glycosylation of selectin ligand CD43 on activated CD8+ T cells but minimally affected the cell surface expression of CD43, activation markers (CD44 and CD69) and cell adhesion molecules (CD11a and CD54). Runx3 KO had a minor effect on lung effector CD8+ T cell death by IAV infection. Our findings indicate that Runx3 differently regulates CD8+ T cell expansion in mLNs and lungs by H1N1 IAV infection. Runx3 is required for CD43 core 2 O-glycosylation on activated CD8+ T cells, and the involved Runx3 signal pathway may mediate CD8+ T cell phenotype for pulmonary generation of CTLs.
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Linfócitos T CD8-Positivos , Subunidade alfa 3 de Fator de Ligação ao Core , Vírus da Influenza A Subtipo H1N1 , Infecções por Orthomyxoviridae , Animais , Camundongos , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Proliferação de Células , Subunidade alfa 3 de Fator de Ligação ao Core/metabolismo , Subunidade alfa 3 de Fator de Ligação ao Core/genética , Glicosilação , Vírus da Influenza A Subtipo H1N1/imunologia , Leucossialina/metabolismo , Pulmão/virologia , Pulmão/metabolismo , Pulmão/imunologia , Pulmão/patologia , Linfonodos/metabolismo , Linfonodos/imunologia , Ativação Linfocitária/imunologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/metabolismo , Infecções por Orthomyxoviridae/virologiaRESUMO
Genetic variants in cell division cycle 42 (CDC42) can manifest with dysmorphic features, autoinflammation, hemophagocytic lymphohistiocytosis, and thrombocytopenia, whereas defective thymopoiesis is a rare disease manifestation. We report a novel CDC42 missense variant (c.46A > G, p.Lys16Glu) resulting in infection and HPV-driven carcinogenesis in the mosaic mother and impaired thymopoiesis and profound T cell lymphopenia in the heterozygous daughter identified through newborn screening for SCID. We found that surface expression of IL-7Rα (CD127) was decreased, consistent with reduced IL-7-induced STAT5 phosphorylation and accelerated apoptotic T cell death. Consistent with the vital role of IL-7 in regulating thymopoiesis, both patients displayed reduced T cell receptor CDR3 repertoires. Moreover, the CDC42 variant prevented binding to the downstream effector, p21-activated kinase (PAK)1, suggesting this impaired interaction to underlie reduced IL-7Rα expression and signaling. Here, we provide the first report of severely compromised thymopoiesis and perturbed IL-7Rα signaling caused by a novel CDC42 variant and presenting with diverging clinical and immunological phenotypes in patients.
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Interleucina-7 , Quinases Ativadas por p21 , Humanos , Recém-Nascido , Apoptose , Interleucina-7/genética , Receptores de Antígenos de Linfócitos T/genética , Transdução de SinaisRESUMO
OBJECTIVE: This study aimed to assess the diagnostic yield of prenatal genetic testing using trio whole exome sequencing (WES) and trio whole genome sequencing (WGS) in pregnancies with fetal anomalies by comparing the results with conventional chromosomal microarray (CMA) analysis. METHODS: A total of 40 pregnancies with fetal anomalies or increased nuchal translucency (NT ≥ 5 mm) were included between the 12th and 21st week of gestation. Trio WES/WGS and CMA were performed in all cases. RESULTS: The trio WES/WGS analysis increased the diagnostic yield by 25% in cases with negative CMA results. Furthermore, all six chromosomal aberrations identified by CMA were independently detected by WES/WGS analysis. In total, 16 out of 40 cases obtained a genetic sequence variant, copy number variant, or aneuploidy explaining the phenotype, resulting in an overall WES/WGS diagnostic yield of 40%. WES analysis provided a more reliable identification of mosaic sequence variants than WGS because of its higher sequencing depth. CONCLUSIONS: Prenatal WES/WGS proved to be powerful diagnostic tools for fetal anomalies, surpassing the diagnostic yield of CMA. They have the potential to serve as standalone methods for prenatal diagnosis. The study highlighted the limitations of WGS in accurately detecting mosaic variants, which is particularly relevant when analyzing chorionic villus samples.
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Sequenciamento do Exoma , Diagnóstico Pré-Natal , Sequenciamento Completo do Genoma , Feminino , Humanos , Gravidez , Diagnóstico Pré-Natal/métodos , Sequenciamento Completo do Genoma/normas , Sequenciamento do Exoma/normas , Análise em Microsséries/normas , Anormalidades Congênitas/genética , Variação Genética/genéticaRESUMO
The emerging evidence has indicated the role of microRNAs (miRNA) in various physiological or pathological processes. Also, documents have suggested that exercise, by affecting miRNA regulation, may enhance burn wound healing. The current study aims to systematically review the role of exercise in regulating miRNAs related to burn wound healing to provide potential therapeutic targets. A comprehensive, systematic search was performed in different international electronic databases, such as Embase, PubMed and Google Scholar search engine, Science Direct, ProQuest and Ovid using keywords extracted from Medical Subject Headings from 2010 to September 2023. The keywords, including 'exercise' AND 'burn wound' AND 'microRNA' and finally, six cases were achieved. Evidence has indicated that exercise may promote the healing of burn wounds by regulating certain miRNAs. Studies have found that exercise regulates the expression of miRNAs such as mir-155, miR-21, let-7a, miR-146a, miR-122 and mir-210 in burn wound tissue, which regulate inflammation and angiogenesis. These findings suggest that miRNAs may play a role in the positive effect of exercise on burn wound healing. However, further research is needed to understand the mechanisms involved fully.
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Sequenciamento do Exoma , Doenças Fetais , Testes Genéticos , Diagnóstico Pré-Natal , Feminino , Humanos , Gravidez , Sequenciamento do Exoma/métodos , Feto/diagnóstico por imagem , Diagnóstico Pré-Natal/métodos , Ultrassonografia Pré-Natal , Doenças Fetais/genética , Testes Genéticos/métodosRESUMO
Spinal cord injury (SCI) is a functional impairment of the spinal cord caused by external forces, accompanied by limb movement disorders and permanent paralysis, which seriously lowers the life quality of SCI patients. Secondary injury caused by inflammation attenuated the therapeutic effects of SCI. Therefore, the exploration of biomarkers associated with the inflammatory response following SCI might provide novel therapy strategy against SCI.SCI rat model was established as previously reported and evaluated by BBB score. The expression of microRNA-24-3p (miR-24-3p) and MAPK-activated protein kinase 2 (MK2) in spinal cord tissues of SCI rats and HAPI cells was analyzed by qRT-PCR. Protein expression of MK2, ionized calcium-binding adapter molecule-1 (Iba-1), tumor necrosis factor-alpha (TNF-α), and interleukin-1ß (IL-1ß) was assessed by western blot assay. The release of inflammatory cytokines TNF-α and IL-1ß was measured by enzyme-linked immunosorbent assay (ELISA). The interaction between miR-24-3p and MK2 was examined by the luciferase reporter system. Basso-Beattie-Bresnahan (BBB) score dramatically reduced in rats following SCI compared with sham rats. Moreover, the expression of miR-24-3p was down-regulated, while MK2 was up-regulated in the spinal cord tissues of SCI rats and LPS-induced microglia cells compared with the corresponding control group. Luciferase reporter system confirmed the interaction between miR-24-3p and MK2. In addition, miR-24-3p upregulation or MK2 knockdown attenuated LPS induced activation of microglial cells and expression of inflammatory cytokine TNF-α and IL-1ß. Besides, we discovered that miR-24-3p regulated inflammation of highly aggressively proliferating immortalized (HAPI) cells by targeting MK2.In our study, we clarified that miR-24-3p repressed inflammation of microglia cells following SCI by regulating MK2, thereby providing promising biomarkers for SCI therapy.
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Inflamação/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , MicroRNAs/metabolismo , Microglia/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Traumatismos da Medula Espinal/metabolismo , Animais , Linhagem Celular Tumoral , Regulação para Baixo/fisiologia , Humanos , Inflamação/etiologia , Peptídeos e Proteínas de Sinalização Intracelular/deficiência , Lipopolissacarídeos/fisiologia , Microglia/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/deficiência , Ratos Sprague-Dawley , Medula Espinal/metabolismo , Traumatismos da Medula Espinal/complicações , Regulação para Cima/fisiologiaRESUMO
BACKGROUND Microglia reside in the spinal cord plays a key role in the onset, progression of post-spinal cord injury (SCI) neuroinflammation. Curcumin has been shown to exhibit diverse anti-inflammatory and anti-tumor activities. The aim of this study was to explore the effect of curcumin on the inflammatory response in lipopolysaccharide (LPS)-activated microglia and its mechanism. MATERIAL AND METHODS The expression levels of phosphorylated-p65 (p-p65), tumor necrosis factor (TNF)-alpha, interleukin (IL)-1ß, and IkappaB kinase ß (IKKß) were examined by western blot assay. MiR-199b-5p expression was detected by quantitative real-time polymerase chain reaction assay. The putative binding sites of miR-199b-5p in IKKß 3'UTR were predicted by bioinformatics, and direct interaction between miR-199b-5p and IKKß was verified by dual-luciferase reporter assay and RNA-immunoprecipitation assay. RESULTS Curcumin significantly suppressed inflammatory response induced by LPS by inactivation of nuclear factor kappa B (NF-kappaB) in microglial cells, as reflected by the decreased levels of p-p65, as well as the pro-inflammatory mediators, including inducible nitric oxide synthase (iNOS), TNF-alpha, and IL-1ß. Moreover, curcumin increased the level of miR-199b-5p and decreased IKKß expression in activated microglial cells. Knockdown of miR-199b-5p or overexpression of IKKß reversed the inhibitory effect of curcumin on inflammatory response and NF-kappaB activation. MiR-199b-5p directly targeted IKKß and suppressed its expression. Silencing of IKKß abolished miR-199b-5p-stimulated inflammatory cytokines production and NF-kappaB activation. CONCLUSIONS Curcumin attenuated neuroinflammation induced by LPS through regulating miR-199b-5p/IKKß/NF-kappaB axis in microglia.
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Curcumina/farmacologia , Microglia/metabolismo , Neuroimunomodulação/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Curcumina/metabolismo , Quinase I-kappa B/metabolismo , Proteínas I-kappa B/metabolismo , Inflamação/patologia , Interleucina-1beta/análise , Lipopolissacarídeos/farmacologia , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Neuroimunomodulação/fisiologia , Óxido Nítrico Sintase Tipo II/metabolismo , Proteínas Serina-Treonina Quinases , Transdução de Sinais/efeitos dos fármacos , Traumatismos da Medula Espinal/patologia , Fator de Transcrição RelA/análise , Fator de Necrose Tumoral alfa/análise , Quinase Induzida por NF-kappaBRESUMO
The freshwater planarian mostly lives in the upper reaches of springs and rivers. Generally, it is realized as a suitable warning indicator of environmental toxicants. The freshwater planarian Dugesia japonica has a powerful regenerative capability and can regenerate a new individual including a complete central nervous system in one week. Rapamycin is an inhibitor of mammalian TORC1 (target of rapamycin complex-1) and used in the treatment of some diseases like cancer, cardiovascular and neurological diseases. However, the roles of rapamycin in the regulation of planarian regeneration remain to be elucidated. In present study, freshwater planarians D. japonica were firstly treated with 1⯵M rapamycin for 18â¯h exposures and the expression patterns of Djtor was analyzed by the whole-mount in situ hybridization (WISH). Our results indicated rapamycin could strongly inhibit Djtor expression in planarian D. japonica and cause asymmetric blastemas and neuronal defects in planarians. Furthermore, knockdown of Djtor gene in planarians using RNA interference resulted in the suppression of downstream autophagy genes. These findings suggested that rapamycin might regulate freshwater planarian regeneration via Djtor signaling pathway.
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Planárias/efeitos dos fármacos , Regeneração/efeitos dos fármacos , Sirolimo/toxicidade , Serina-Treonina Quinases TOR/metabolismo , Poluentes Químicos da Água/toxicidade , Animais , Sistema Nervoso Central/efeitos dos fármacos , Neurônios , Planárias/genética , Planárias/crescimento & desenvolvimento , Planárias/metabolismo , Interferência de RNA , Regeneração/fisiologia , Transdução de Sinais , Serina-Treonina Quinases TOR/genéticaRESUMO
BACKGROUND: The prognosis of lung cancer is very poor and hence new therapeutic strategies are urgently desired. In this study, we searched for efficacious Smac mimetic-based combination therapies with biomarkers to predict responses for non-small cell lung cancer (NSCLC). METHODS: NSCLC cell lines and normal human alveolar epithelial cells were treated with Smac mimetics plus IFNγ or other agonists and cell viabilities were assessed by MTS assay, cell counting, flow cytometry and cell colony assay. Western blot analysis was performed to assess the cleavage (activation) of caspases and expression of signaling molecules. Caspase activity was determined to verify caspase activation. The pathways involved in NSCLC cell death were investigated using specific inhibitors. RESULTS: We found that IFNγ could cooperate with various Smac mimetics to trigger a profound apoptosis in a number of NSCLC cell lines that are competent for IFNγ signaling (i.e. expressing IFNγ receptor-1 and STAT1) but have low expression levels of inhibitor of apoptosis proteins survivin and livin without harming normal human lung epithelial cells. IFNγ co-treatment with a novel class dimeric Smac mimetic AZD5582 eradicated NSCLC cell colony formation. Unlike IFNγ, IFNα, IFNλ, TNFα, or TRAIL alone or plus AZD5582 had minor effects on NSCLC cell viability. IFNγ/AZD5582-induced cell death in NSCLC cells was independent of TNFα autocrine but relied on apoptosis mediated by JAK kinase, caspase 8 and RIPK1 pathways. CONCLUSION: Our results indicate that IFNγ and Smac mimetics can synergize to induce apoptosis of NSCLC cells and suggest that IFNγ and Smac mimetic regimen may be a novel and efficacious apoptosis targeted therapy with biomarkers to predict responses for NSCLC cells.
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OBJECTIVE: To investigate human papilloma virus (HPV) infection in the male genital tract and its risk factors in some rural areas of Jiangsu Province. METHODS: This study included 398 men from six rural areas in Jiangsu Province, whose female partners, based on the results of cervical cytological examination, were divided into a normal (n = 104), a cervical intraepithelial neoplasia grade â (CIN-â , n = 100), a CIN-â ¡ (n = 95), and a CIN-â ¢ group (n = 99). We examined the male subjects for genital warts and other lesions, collected urethral swab samples for HPV detection, and obtained their sociodemographic data by questionnaire investigation. RESULTS: No megascopic lesions were observed in the genitals of the 398 participants. The total prevalence rate of HPV infection was 11.31% and that of high-risk HPV was 8.54%. Logistic regression analysis showed that daily cleaning of the genitals significantly decreased the risk of HPV infection (OR = 3.030, P = 0.003). CONCLUSIONS: There is a relatively high prevalence rate of recessive infection of genital HPV among the seemingly healthy males in the rural area of Jiangsu Province. Daily cleaning of the genitals may be a protective measure against HPV infection.
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OBJECTIVE: To investigate the correlation of the single nucleotide polymorphism (SNP) rs1042522 of the tumor protein p53 (TP53) gene with the risk of male infertility. METHODS: This caseîcontrol study included 380 male patients with idiopathic infertility and 398 normal fertile men as controls from the Nanjing area. We genotyped the SNP rs1042522 of the TP53 gene by Sequence Mass Array and analyzed the correlation of the SNP with male infertility using the logistic regression model. RESULTS: Compared with the normal controls, the patients with idiopathic infertility showed significantly decreased sperm concentration (ï¼»77.34±49.24ï¼½ vs ï¼»13.13±24.96ï¼½ ×106/ml), percentage of progressively motile sperm (ï¼»42.55±9.57ï¼½ vs ï¼»10.38±5.57ï¼½%), serum testosterone level (ï¼»14.07±5.36ï¼½ vs ï¼»11.89±4.50ï¼½ nmol/L), and follicleîstimulating hormone level (ï¼»16.80±18.20ï¼½ vs ï¼»4.55±7.17ï¼½ U/L) (P < 0.05) but no statistically significant differences in other parameters. No correlation was observed between the SNP frequencies and male infertility and similar results were found in the subgroups of the cases. CONCLUSIONS: SNP rs1042522 of the TP53 gene is not significantly correlated with the risk of male infertility.
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Genes p53/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Contagem de Espermatozoides , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Infertilidade Masculina/sangue , Modelos Logísticos , Masculino , Motilidade dos Espermatozoides , Testosterona/análogos & derivados , Testosterona/sangueRESUMO
OBJECTIVE: To investigate the correlation of the single nucleotide polymorphism (SNP) rs4880 of the superoxide dismutase 2 (SOD2) gene with the risk of male infertility. METHODS: This caseîcontrol study included 519 male patients with idiopathic infertility (aged 19ï¼40 ï¼»28.93±4.93ï¼½ years) in the case group and 338 fertile men (aged 19ï¼40 ï¼»28.40±4.25ï¼½ years) in the control group. We collected the clinical data, genotyped the SNP rs4880 of the SOD2 gene by Sequenom Mass Array, and analyzed the association of different genotypes with male infertility using the logistic regression model. RESULTS: Statically significant differences were observed between the case and control groups in the level of follicleîstimulating hormone (FSH) (ï¼»4.72±2.51ï¼½ vs ï¼»15.65±17.24ï¼½ U/L, P< 0.01), the percentage of progressively mobile sperm (ï¼»9.12±13.5ï¼½ vs ï¼»41.95±9.03ï¼½%, P< 0.01), and sperm concentration (ï¼»12.95±24.38ï¼½ vs ï¼»72.88±45.60ï¼½ ×106/ml, P< 0.01), but not in other parameters. No correlation was found between male infertility and the heterozygous genotype TC (OR = 0.90, 95% CI: 0.65ï¼1.25, P = 0.516) or the homozygous genotype CC (OR=1.49, 95% CI: 0.38ï¼5.81, P = 0.566) as compared with the wild genotype TT, and similar results were obtained in the analysis of the subgroups. CONCLUSIONS: The SNP rs4880 of the SOD2 gene was not correlated with male infertility, which, however, is to be supported by further studies with larger samples from more areas.
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Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Superóxido Dismutase/genética , Adulto , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Predisposição Genética para Doença , Genótipo , Heterozigoto , Humanos , Modelos Logísticos , Masculino , Nucleotídeos/genética , Motilidade dos Espermatozoides , Adulto JovemRESUMO
Viral respiratory tract infections are the most common illness in humans. Infection of the respiratory viruses results in accumulation of viral replicative double-stranded RNA (dsRNA), which is one of the important components of infecting viruses for the induction of lung epithelial cell apoptosis and innate immune response, including the production of interferon (IFN). In the present study, we have investigated the regulation of dsRNA-induced airway epithelial cell apoptosis by IFN. We found that transcription factor Runx3 was strongly induced by type-II IFNγ, slightly by type-III IFNλ, but essentially not by type-I IFNα in airway epithelial cells. IFNγ-induced expression of Runx3 was predominantly mediated by JAK-STAT1 pathway and partially by NF-κB pathway. Interestingly, Runx3 can be synergistically induced by IFNγ with a synthetic analog of viral dsRNA polyinosinic-polycytidylic acid [poly(I:C)] or tumor necrosis factor-α (TNFα) through both JAK-STAT1 and NF-κB pathways. We further found that dsRNA poly(I:C)-induced apoptosis of airway epithelial cells was mediated by dsRNA receptor toll-like receptor 3 (TLR3) and was markedly augmented by IFNγ through the enhanced expression of TLR3 and subsequent activation of both extrinsic and intrinsic apoptosis pathways. Last, we demonstrated that upregulation of Runx3 by IFNγ promoted TLR3 expression, thus amplifying the dsRNA-induced apoptosis in airway epithelial cells. These novel findings indicate that IFNγ promotes dsRNA-induced TLR3-dependent apoptosis via upregulation of transcription factor Runx3 in airway epithelial cells. Findings from our study may provide new insights into the regulation of airway epithelial cell apoptosis by IFNγ during viral respiratory tract infection.
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Apoptose/efeitos dos fármacos , Subunidade alfa 3 de Fator de Ligação ao Core/metabolismo , Células Epiteliais/metabolismo , Interferon gama/farmacologia , Pulmão/citologia , RNA de Cadeia Dupla/farmacologia , Receptor 3 Toll-Like/metabolismo , Regulação para Cima/efeitos dos fármacos , Linhagem Celular , Células Epiteliais/efeitos dos fármacos , Humanos , Janus Quinases/metabolismo , NF-kappa B/metabolismo , Poli I-C/farmacologia , Fator de Transcrição STAT1/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Female C57BL/6J mice were fed a regular low-fat diet or high-fat diets combined with either high or low protein-to-sucrose ratios during their entire lifespan to examine the long-term effects on obesity development, gut microbiota, and survival. Intake of a high-fat diet with a low protein/sucrose ratio precipitated obesity and reduced survival relative to mice fed a low-fat diet. By contrast, intake of a high-fat diet with a high protein/sucrose ratio attenuated lifelong weight gain and adipose tissue expansion, and survival was not significantly altered relative to low-fat-fed mice. Our findings support the notion that reduced survival in response to high-fat/high-sucrose feeding is linked to obesity development. Digital gene expression analyses, further validated by qPCR, demonstrated that the protein/sucrose ratio modulated global gene expression over time in liver and adipose tissue, affecting pathways related to metabolism and inflammation. Analysis of fecal bacterial DNA using the Mouse Intestinal Tract Chip revealed significant changes in the composition of the gut microbiota in relation to host age and dietary fat content, but not the protein/sucrose ratio. Accordingly, dietary fat rather than the protein/sucrose ratio or adiposity is a major driver shaping the gut microbiota, whereas the effect of a high-fat diet on survival is dependent on the protein/sucrose ratio.
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Dieta com Restrição de Gorduras , Proteínas Alimentares/farmacocinética , Sacarose Alimentar/farmacocinética , Microbioma Gastrointestinal/fisiologia , Obesidade/metabolismo , Taxa de Sobrevida , Animais , Proteínas Alimentares/administração & dosagem , Proteínas Alimentares/efeitos adversos , Sacarose Alimentar/efeitos adversos , Feminino , Estudos Longitudinais , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/etiologiaRESUMO
OBJECTIVE: In this study, we evaluated the diagnostic efficiency of six recombinant proteins for the serodiagnosis of Lyme borreliosis (LB) and screened out the appropriate antigens to support the production of a Chinese clinical ELISA (enzyme-linked immunosorbent assay) kit for LB. METHODS: Six recombinant antigens, Fla B.g, OspC B.a, OspC B.g, P39 B.g, P83 B.g, and VlsE B.a, were used for ELISA to detect serum antibodies in LB, syphilis, and healthy controls. The ELISA results were used to generate receiver operating characteristic (ROC) curves, and the sensitivity and specificity of each protein was evaluated. All recombinant proteins were evaluated and screened by using logistic regression models. RESULTS: Two IgG (VlsE and OspC B.g) and two IgM (OspC B.g and OspC B.a) antigens were left by the logistic regression model screened. VlsE had the highest specificity for syphilis samples in the IgG test (87.7%, P<0.05). OspC B.g had the highest diagnostic value in the IgM test (AUC=0.871). Interactive effects between OspC B.a and Fla B.g could reduce the specificity of the ELISA. CONCLUSION: Three recombinant antigens, OspC B.g, OspC B.a, and VlsE B.a, were useful for ELISAs of LB. Additionally, the interaction between OspC B.a and Fla B.g should be examined in future research.
Assuntos
Proteínas de Bactérias/análise , Ensaio de Imunoadsorção Enzimática/veterinária , Doença de Lyme/diagnóstico , Testes Sorológicos/veterinária , Antígenos de Bactérias/sangue , China , Proteínas Recombinantes/análise , Sensibilidade e EspecificidadeRESUMO
Thirty surface sediments and three sediment cores were collected from mangrove wetlands in the Pearl River Estuary of South China to investigate the spatial and vertical distribution of Dechlorane Plus (DP). DP concentrations in the mangrove surface sediments ranged from 0.0130 to 1.504 ng/g dry weight (dw). DP concentrations in sediments from Shenzhen were significantly greater than those from Guangzhou and Zhuhai. Anti-Cl11-DP, the dechlorinated product of anti-DP, was also detected in the mangrove sediments with concentrations ranged from not detected to 0.0198 ng/g dw. Significant positive relationship between anti-Cl11-DP and anti-DP levels was observed in the mangrove sediments, suggesting that photo and/or microbial degradation of anti-DP might occur in the sediments. The f anti values in the mangrove sediments were close to those in the technical DP products, suggesting that stereoselective enrichment of anti-DP may not exist in the mangrove sediments. DP concentrations in the mangrove sediment cores generally showed an increasing trend from the bottom to top layers. This is the first study to report the occurrence of DP and its degradation product in the mangrove wetlands.
Assuntos
Monitoramento Ambiental , Sedimentos Geológicos/química , Hidrocarbonetos Clorados/análise , Compostos Policíclicos/análise , Poluentes Químicos da Água/análise , Áreas Alagadas , ChinaRESUMO
A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients' serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.