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BACKGROUND: In several countries, thyroid dyshormonogenesis is more common than thyroid dysgenesis in patients with congenital hypothyroidism (CH). However, known pathogenic genes are limited to those directly involved in hormone biosynthesis. The aetiology and pathogenesis of thyroid dyshormonogenesis remain unknown in many patients. METHODS: To identify additional candidate pathogenetic genes, we performed next-generation sequencing in 538 patients with CH and then confirmed the functions of the identified genes in vitro using HEK293T and Nthy-ori 3.1 cells, and in vivo using zebrafish and mouse model organisms. RESULTS: We identified one pathogenic MAML2 variant and two pathogenic MAMLD1 variants that downregulated canonical Notch signalling in three patients with CH. Zebrafish and mice treated with N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butylester, a γ-secretase inhibitor exhibited clinical manifestations of hypothyroidism and thyroid dyshormonogenesis. Through organoid culture of primary mouse thyroid cells and transcriptome sequencing, we demonstrated that Notch signalling within thyroid cells directly affects thyroid hormone biosynthesis rather than follicular formation. Additionally, these three variants blocked the expression of genes associated with thyroid hormone biosynthesis, which was restored by HES1 expression. The MAML2 variant exerted a dominant-negative effect on both the canonical pathway and thyroid hormone biosynthesis. MAMLD1 also regulated hormone biosynthesis through the expression of HES3, the target gene of the non-canonical pathway. CONCLUSIONS: This study identified three mastermind-like family gene variants in CH and revealed that both canonical and non-canonical Notch signalling affected thyroid hormone biosynthesis.
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Hipotiroidismo Congénito , Animales , Humanos , Ratones , Hipotiroidismo Congénito/genética , Proteínas de Unión al ADN/genética , Células HEK293 , Mutación , Proteínas Nucleares/genética , Hormonas Tiroideas/genética , Transactivadores/genética , Factores de Transcripción/genética , Pez CebraRESUMEN
Nonsyndromic cleft palate only (NSCP) is a common congenital malformation worldwide. In this study, we report a three-generation pedigree with NSCP following the autosomal-dominant pattern. Whole-exome sequencing and Sanger sequencing revealed that only the frameshift variant c.1012dupG [p. E338Gfs*26] in PARD3 cosegregated with the disease. In zebrafish embryos, ethmoid plate patterning defects were observed with PARD3 ortholog disruption or expression of patient-derived N-terminal truncating PARD3 (c.1012dupG), which implicated PARD3 in ethmoid plate morphogenesis. PARD3 plays vital roles in determining cellular polarity. Compared with the apical distribution of wild-type PARD3, PARD3-p. E338Gfs*26 mainly localized to the basal membrane in 3D-cultured MCF-10A epithelial cells. The interaction between PARD3-p. E338Gfs*26 and endogenous PARD3 was identified by LC-MS/MS and validated by co-IP. Immunofluorescence analysis showed that PARD3-p. E338Gfs*26 substantially altered the localization of endogenous PARD3 to the basement membrane in 3D-cultured MCF-10A cells. Furthermore, seven variants, including one nonsense variant and six missense variants, were identified in the coding region of PARD3 in sporadic cases with NSCP. Subsequent analysis showed that PARD3-p. R133*, like the insertion variant of c.1012dupG, also changed the localization of endogenous full-length PARD3 and that its expression induced abnormal ethmoid plate morphogenesis in zebrafish. Based on these data, we reveal PARD3 gene variation as a novel candidate cause of nonsyndromic cleft palate only.
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Labio Leporino , Fisura del Paladar , Animales , Cromatografía Liquida , Labio Leporino/genética , Fisura del Paladar/genética , Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple , Espectrometría de Masas en Tándem , Pez Cebra/genéticaRESUMEN
BACKGROUND AND OBJECTIVE: Type 2 diabetes (T2D)-associated periodontitis is severe and refractory in many cases. Considered an inflammatory disease, T2D predisposes to periodontitis by increasing whole-body inflammation. One mechanism of increased inflammation is thatT2D is mediated by loss of production or function of the anti-inflammatory hormone adiponectin. In our previous report, AdipoRon, an adiponectin receptor agonist, and AdipoAI, a newly discovered, more specific agonist, attenuated T2D-associated inflammation by inhibiting osteoclastogenesis and LPS-induced endotoxemia. Autophagy plays an important role during osteoclast differentiation and function. The impact of AdipoAI on osteoclast function and autophagy involved in osteoclastogenesis is not known. Here, we compare AdipoRon and AdipoAI potency, side effects and mechanism of action in T2D-associated periodontitis. METHODS: The RAW 264.7 cell line was used for in vitro studies. We analyzed the potential cytotoxicity of AdipoAI using the CCK-8 assay. The anti-osteoclastogenic potential of AdipoAI was studied by real-time qPCR and tartrate-resistant acid phosphatase staining. The actions of AdipoAI involved in autophagy were tested by real-time qPCR, western blot and immunofluorescence staining. In the diet-induced obesity model of T2D, we investigated the impact of AdipoAI on fasting blood glucose, alveolar bone loss, and gingival inflammation in mice with experimental periodontitis. RESULTS: AdipoRon inhibited osteoclastogenesis and AdipoAI inhibited osteoclastogenesis at lower doses than AdipoRon without any cytotoxicity. In DIO mice with experimental periodontitis, AdipoAI reduced mouse body weight in 14 days, reducing fasting glucose levels, alveolar bone destruction, osteoclast number along the alveolar bone surface, and decreased the expression of pro-inflammatory factors in periodontal tissues. AdipoAI and AdipoRon also enhanced LC3A/B expression when cultured with RANKL.3-Methyladenine, a known autophagy inhibitor, decreased LC3A/B expression and reversed the inhibition of osteoclastogenesis during AdipoAI treatment. CONCLUSIONS: Our results demonstrate that AdipoAI ameliorates the severity of T2D-associated periodontitis by enhancing autophagy in osteoclasts at lower doses than AdipoRon without demonstrable side effects. Thus, AdipoAI has pharmaceutical potential for treating diabetes-associated periodontal disease.
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Pérdida de Hueso Alveolar , Diabetes Mellitus Tipo 2 , Periodontitis , Adiponectina , Pérdida de Hueso Alveolar/tratamiento farmacológico , Pérdida de Hueso Alveolar/metabolismo , Pérdida de Hueso Alveolar/prevención & control , Animales , Autofagia , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Ratones , Osteoclastos , Periodontitis/tratamiento farmacológico , Periodontitis/metabolismo , Ligando RANK/metabolismo , Receptores de Adiponectina/agonistas , Receptores de Adiponectina/metabolismo , Receptores de Adiponectina/uso terapéuticoRESUMEN
KEY MESSAGE: A total of 33 additive stem rot QTLs were identified in peanut genome with nine of them consistently detected in multiple years or locations. And 12 pairs of epistatic QTLs were firstly reported for peanut stem rot disease. Stem rot in peanut (Arachis hypogaea) is caused by the Sclerotium rolfsii and can result in great economic loss during production. In this study, a recombinant inbred line population from the cross between NC 3033 (stem rot resistant) and Tifrunner (stem rot susceptible) that consists of 156 lines was genotyped by using 58 K peanut single nucleotide polymorphism (SNP) array and phenotyped for stem rot resistance at multiple locations and in multiple years. A linkage map consisting of 1451 SNPs and 73 simple sequence repeat (SSR) markers was constructed. A total of 33 additive quantitative trait loci (QTLs) for stem rot resistance were detected, and six of them with phenotypic variance explained of over 10% (qSR.A01-2, qSR.A01-5, qSR.A05/B05-1, qSR.A05/B05-2, qSR.A07/B07-1 and qSR.B05-1) can be consistently detected in multiple years or locations. Besides, 12 pairs of QTLs with epistatic (additive × additive) interaction were identified. An additive QTL qSR.A01-2 also with an epistatic effect interacted with a novel locus qSR.B07_1-1 to affect the percentage of asymptomatic plants in a row. A total of 193 candidate genes within 38 stem rot QTLs intervals were annotated with functions of biotic stress resistance such as chitinase, ethylene-responsive transcription factors and pathogenesis-related proteins. The identified stem rot resistance QTLs, candidate genes, along with the associated SNP markers in this study, will benefit peanut molecular breeding programs for improving stem rot resistance.
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Arachis/genética , Mapeo Cromosómico , Resistencia a la Enfermedad/genética , Epistasis Genética , Genes de Plantas , Enfermedades de las Plantas/genética , Tallos de la Planta/microbiología , Sitios de Carácter Cuantitativo/genética , Ascomicetos/fisiología , Ligamiento Genético , Fenotipo , Enfermedades de las Plantas/microbiologíaRESUMEN
Recent studies have demonstrated that aberrant long non-coding RNAs (lncRNAs) expression are suggested to be closely associated with multiple human diseases, lung cancer included. However, the roles of lncRNAs in lung cancer are not well understood. In this study, we used microarrays to investigate the aberrantly expressed lncRNAs in the mouse lung adenocarcinoma with P53 knockout and the KrasG12D mutation. Results revealed that 6424 lncRNAs were differentially expressed (≥ 2-fold change, P < .05). Two hundred and ten lncRNAs showed more than 8-fold change and conserved across human and were further analysed in the primary mouse lung adenocarcinoma KP cells, which were isolated from the p53 knockout and the KrasG12D mutation mice. Among all the 210 lncRNAs, 11 lncRNAs' expression was regulated by P53, 33 lncRNAs by KRAS and 13 lncRNAs by hypoxia in the primary KP cells, respectively. NONMMUT015812, which was remarkably up-regulated in the mouse lung adenocarcinoma and negatively regulated by the P53 re-expression, was detected to analyse its cellular function. Results showed that knockdown of NONMMUT015812 by shRNAs decreased proliferation and migration abilities of KP cells. Among those aberrantly expressed lncRNAs in the mouse lung adenocarcinoma, NONMMUT015812 was a potential oncogene.
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Adenocarcinoma del Pulmón/genética , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Mutación/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , ARN Largo no Codificante/genética , Proteína p53 Supresora de Tumor/genética , Adenocarcinoma del Pulmón/patología , Animales , Secuencia de Bases , Hipoxia de la Célula/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Células HEK293 , Humanos , Neoplasias Pulmonares/patología , Ratones Noqueados , Oncogenes , ARN Largo no Codificante/metabolismoRESUMEN
Atom-scale modulation of electronic regulation in nonprecious-based electrocatalysts is promising for efficient catalytic activities. Here, hierarchically hollow VOOH nanostructures are rationally constructed by partial iron substitution and systematically investigated for electrocatalytic water splitting. Benefiting from the hierarchically stable scaffold configuration, highly electrochemically active surface area, the synergistic effect of the active metal atoms, and optimal adsorption energies, the 3% Fe (mole ratio) substituted electrocatalyst (VOOH-3Fe) exhibits a low overpotential of 90 and 195 mV at 10 mA cm-2 for hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) in alkaline media, respectively, superior than the other samples with a different substituted ratio. To the best of current knowledge, 195 mV overpotential at 10 mA cm-2 is the best value reported for V or Fe (oxy)hydroxide-based OER catalysts. Moreover, the electrolytic cell employing the VOOH-3Fe electrode as both the cathode and anode exhibits a cell voltage of 0.30 V at 10 mA cm-2 with a remarkable stability over 60 h. This work heralds a new pathway to design efficient bifunctional catalysts toward overall water splitting.
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PURPOSE: To detect the expression of VEGF and EGFR in peripheral blood and cancer tissues of patients with renal cell carcinoma (RCC), and to explore the correlations with clinical stage, pathological grade and prognosis of disease. METHODS: A total of 64 patients with RCC who were diagnosed and treated from June 2016 to August 2017 in our hospital were enrolled. Patients were divided into different clinical stages and pathological grades, and ELISA and immunohistochemistry were used to detect the expression of VEGF and EGFR in peripheral blood. Peripheral blood was also taken from 24 healthy individuals to serve as control group. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of VEGF and EGFR in RCC tissues and paracancer tissues. All patients were followed up after discharge to record their survival. RESULTS: Significant differences in the expression levels of VEGF and EGFR were found between stage III and IV (p<0.05), but not between stage I and II. Expressions level of VEGF and EGFR in serum of well-differentiated, moderatelydifferentiated, and poorly-differentiated RCC were all higher than those in the healthy control group, and significant differences were found between different pathological grades (p<0.05). Patients with higher expression levels of VEGF and EGFR showed shorter survival compared to patients with lower expression levels (p<0.05). CONCLUSION: VEGF and EGFR in peripheral blood can be used as one of the effective indicators of prognosis of RCC. Our study provided reference for clinical treatment and prediction of prognosis of RCC.
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Carcinoma de Células Renales/sangre , Carcinoma de Células Renales/patología , Neoplasias Renales/sangre , Neoplasias Renales/patología , Factor A de Crecimiento Endotelial Vascular/sangre , Adulto , Anciano , Carcinoma de Células Renales/mortalidad , Estudios de Casos y Controles , Receptores ErbB/biosíntesis , Receptores ErbB/sangre , Femenino , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Neoplasias Renales/mortalidad , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Estadificación de Neoplasias , Pronóstico , Factor A de Crecimiento Endotelial Vascular/biosíntesisRESUMEN
Several mutations in the IRF6 gene have been identified as a causative link to VWS. In this investigation, whole-exome sequencing (WES) and Sanger sequencing of a three-generation pedigree with an autosomal-dominant inheritance pattern affected by VWS identified a unique stop-gain mutation-c.748C>T:p.R250X-in the IRF6 gene that co-segregated exclusively with the disease phenotype. Immunofluorescence analysis revealed that the IRF6-p.R250X mutation predominantly shifted its localization from the nucleus to the cytoplasm. WES and protein interaction analyses were conducted to understand this mutation's role in the pathogenesis of VWS. Using LC-MS/MS, we found that this mutation led to a reduction in the binding of IRF6 to histone modification-associated proteins (NAA10, SNRPN, NAP1L1). Furthermore, RNA-seq results show that the mutation resulted in a downregulation of TGFß2-AS1 expression. The findings highlight the mutation's influence on TGFß2-AS1 and its subsequent effects on the phosphorylation of SMAD2/3, which are critical in maxillofacial development, particularly the palate. These insights contribute to a deeper understanding of VWS's molecular underpinnings and might inform future therapeutic strategies.
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Background: Congenital hypothyroidism (CH) is the most common neonatal metabolic disorder. In patients with CH in China, thyroid dyshormonogenesis is more common than thyroid dysgenesis; however, the genetic causes of CH due to thyroid dyshormonogenesis remain largely unknown. Therefore, we aimed at identifying novel candidate causative genes for CH. Methods: To identify novel CH candidate genes, a total of 599 patients with CH were enrolled and next-generation sequencing was performed. The functions of the identified variants were confirmed using HEK293T and FTC-133 cell lines in vitro and in a mouse model organism in vivo. Results: Three pathogenic contactin 6 (CNTN6) variants were identified in two patients with CH. Pedigree analysis showed that CH caused by CNTN6 variants was inherited in an autosomal recessive pattern. The CNTN6 gene was highly expressed in the thyroid in humans and mice. Cntn6 knockout mice presented with thyroid dyshormonogenesis and CH due to the decreased expression of crucial genes for thyroid hormone biosynthesis (Slc5a5, Tpo, and Duox2). All three CNTN6 variants resulted in the blocking of the release of the Notch intracellular domain, which could not translocate into the nucleus, impaired NOTCH1 transcriptional activity, and decreased expression of SLC5A5, TPO, and DUOX2. Further, we found that DTX1 was required for CNTN6 to promote thyroid hormone biosynthesis through Notch signaling. Conclusions: This study demonstrated that CNTN6 is a novel causative gene for CH through the mediation of thyroid hormone biosynthesis via Notch signaling, which provides new insights into the genetic background and mechanisms involved in CH and thyroid dyshormonogenesis.
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Hipotiroidismo Congénito , Humanos , Animales , Ratones , Hipotiroidismo Congénito/genética , Oxidasas Duales/genética , Células HEK293 , Mutación , Yoduro Peroxidasa/genética , Hormonas Tiroideas , Contactinas/genéticaRESUMEN
Background: Genetic defects in the human thyroid-stimulating hormone (TSH) receptor (TSHR) gene can cause congenital hypothyroidism (CH). However, the biological functions and comprehensive genotype-phenotype relationships for most TSHR variants associated with CH remain unexplored. We aimed to identify TSHR variants in Chinese patients with CH, analyze the functions of the variants, and explore the relationships between TSHR genotypes and clinical phenotypes. Methods: In total, 367 patients with CH were recruited for TSHR variant screening using whole-exome sequencing. The effects of the variants were evaluated by in-silico programs such as SIFT and polyphen2. Furthermore, these variants were transfected into 293T cells to detect their Gs/cyclic AMP and Gq/11 signaling activity. Results: Among the 367 patients with CH, 17 TSHR variants, including three novel variants, were identified in 45 patients, and 18 patients carried biallelic TSHR variants. In vitro experiments showed that 10 variants were associated with Gs/cyclic AMP and Gq/11 signaling pathway impairment to varying degrees. Patients with TSHR biallelic variants had lower serum TSH levels and higher free triiodothyronine and thyroxine levels at diagnosis than those with DUOX2 biallelic variants. Conclusions: We found a high frequency of TSHR variants in Chinese patients with CH (12.3%), and 4.9% of cases were caused by TSHR biallelic variants. Ten variants were identified as loss-of-function variants. The data suggest that the clinical phenotype of CH patients caused by TSHR biallelic variants is relatively mild. Our study expands the TSHR variant spectrum and provides further evidence for the elucidation of the genetic etiology of CH.
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Hipotiroidismo Congénito , Humanos , China , Hipotiroidismo Congénito/diagnóstico , Hipotiroidismo Congénito/genética , AMP Cíclico , Oxidasas Duales/genética , Mutación , Fenotipo , Receptores de Tirotropina/genética , TirotropinaRESUMEN
The effectiveness of SARS-CoV-2 vaccines varies among individuals. During the COVID-19 global pandemic, SARS-CoV-2 infection showed significant Th1 characteristics, suggesting that the immune disorder and production of SARS-CoV-2 antibodies may be related to Th1/Th2 bias. However, the molecular mechanisms underlying Th1/Th2 bias effects on host immune responses to viruses remain unclear. In this study, the top three subjects with the highest and lowest changes in anti-SARS-CoV-2 antibodies after receiving three doses of SARS-CoV-2 vaccination were selected and defined as the elevated group (E) and the control group (C), respectively. Peripheral blood was collected, single-cell sequencing was performed before and after the third dose of the SARS-CoV-2 vaccine, and the changes in T cell clusters were analyzed. Compared with the C group, the Treg pre-vaccination proportion was lower in E, while the post-vaccination proportion was higher, suggesting that Tregs may be crucial in this process. Differential analysis results of Tregs between the two groups revealed that differentially expressed genes (DEGs) were significantly enriched in the IL4 pathway. Correlation analysis between DEGs and serum antibody showed that the expression of NR4A2, SOCS1, and SOCS3 in Tregs was significantly correlated with serum antibodies, suggesting that the immune response in E group changed to Th2 bias, thereby promoting host humoral immune responses. On the other hand, antibody-related genes SOCS1 and NR4A2, as well as lnc-RNA MALAT1 and NEAT1, were highly expressed in the CD4-MALAT1 subclusters. In summary, our study revealed that Th2 bias promotes humoral immune responses in humans by increasing SOCS1 in T cells after SARS-CoV-2 vaccination. Moreover, NR4A2, SOCS1, MALAT1, and NEAT1 were identified as the potential key biomarkers or treatment targets for enhanced SARS-CoV-2 antibody production by influencing the Th1/Th2 balance in T cells. Our findings have important implications for population stratification and tailored therapeutics for more effective SARS-CoV-2 vaccines.
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INTRODUCTION: Congenital hypothyroidism (CH), the most common neonatal endocrine disorder world-wide, can be caused by variants in the thyroid peroxidase (TPO) gene. This study aimed to identify TPO variants in Chinese patients with CH, analyze their impact on TPO function, and establish relationships between TPO genotypes and clinical characteristics. METHODS: A total of 328 patients with CH were screened for TPO variants by performing whole exome sequencing. The function of the detected TPO variants was investigated via transfection assays in vitro. The pathogenic effect of five novel variants was further assessed in silico. RESULTS: Among 328 patients with CH, 19 TPO variants, including six novel ones, were identified in 43 patients. Eighteen patients (5.5%) carried biallelic TPO variants. In vitro experiments showed that TPO activity was impaired to varying degrees in 17 variants. Furthermore, we determined that a residual TPO enzyme activity threshold of 15% may serve as a criterion for differentiating CH severity. CONCLUSIONS: According to our study, the prevalence of TPO variants among Chinese patients with CH was 13.1 %. Five novel variants led to impaired TPO function by altering its structure or by affecting its expression or cellular localization, which should result in impaired thyroid hormone synthesis.
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INTRODUCTION: Urethral trauma is often associated with erectile dysfunction (ED). Reconstructive surgery is complex and may impact negatively on sexual function. AIM: The aim of this article is to investigate ED in patients with pelvic fracture urethral distraction defects (PFUDD) who underwent urethroplasty, and efficacy of treatment with sildenafil citrate. MAIN OUTCOME MEASURES: A total of 41 patients with urethral stricture who suffered from PFUDD were assessed to exclude systemic diseases that may cause ED, such as hypertension, diabetes mellitus, heart disease, and chronic liver disease. The International Index of Erectile Function-5 was used as an evaluation tool. Assessments were made at three time points: the time of admission, two weeks after urethroplasty, and 3 months post-treatment with sildenafil. METHODS: Pharmacopenile duplex ultrasonography was used to examine blood flow of the cavernosum in order to distinguish arterial ED, venous ED, and nonvascular ED. All patients were treated with oral sildenafil, 100 mg once daily, three times a week, for 3 months. RESULTS: The incidence of ED following injury was 95.12%. There were no significant changes in scores following surgery. However, sildenafil had a success rate of approximately 81%, which appeared to be independent of age. Drug treatment seemed most effective for those with less severe ED at the outset. There was no significant difference in scores post-treatment between those who had vascular and nonvascular ED. Overall, the incidence of side effects due to sildenafil was 19.5%. CONCLUSIONS: Urethral trauma is frequently associated with ED. Sildenafil citrate is useful in the drug treatment of ED in these patients and appears to be well-tolerated.
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Disfunción Eréctil/tratamiento farmacológico , Inhibidores de Fosfodiesterasa 5/uso terapéutico , Piperazinas/uso terapéutico , Sulfonas/uso terapéutico , Estrechez Uretral/cirugía , Adolescente , Adulto , Factores de Edad , Disfunción Eréctil/complicaciones , Humanos , Masculino , Persona de Mediana Edad , Pene/irrigación sanguínea , Pene/diagnóstico por imagen , Inhibidores de Fosfodiesterasa 5/efectos adversos , Piperazinas/efectos adversos , Purinas/efectos adversos , Purinas/uso terapéutico , Procedimientos de Cirugía Plástica/métodos , Índice de Severidad de la Enfermedad , Citrato de Sildenafil , Sulfonas/efectos adversos , Resultado del Tratamiento , Ultrasonografía , Uretra/cirugía , Estrechez Uretral/complicaciones , Adulto JovenRESUMEN
INTRODUCTION: To investigate factors correlated with erectile dysfunction (ED) in patients with traumatic urethral strictures undergoing end-to-end anastomotic urethroplasty (AU). MATERIALS AND METHODS: Between January 2010 and January 2011, 41 patients with urethral strictures resulting from pelvic fracture urethral distraction defects underwent end-to-end AU. The abridged International Index of Erectile Function (IIEF-5) was used to subjectively assess erectile function at admission and 2 weeks postoperatively. RESULTS: Pre- and post-injury IIEF-5 scores differed significantly (23.54 ± 1.45 versus 10.02 ± 3.57; p < 0.0001), but pre and postoperative scores did not (10.02 ± 3.57 versus 9.29 ± 4.14; p = 0.1560). Erectile function declined in all patients after injury and was postoperatively unchanged in 56.10%. Pre- and post-injury scores differed significantly in all ages, stricture location and length groups, but did not change postoperatively. Urethral injury resulted in varying degrees of ED. IIEF-5 scores declined significantly postoperatively in patients with mild/mild-moderate ED (13.86 ± 1.88 versus 11.43 ± 3.37; p = 0.0202), but were unchanged in patients with moderate/severe ED. Vascular ED was predominant (63.41%), and erectile function was better in patients with non-vascular ED than in those with arterial/venous ED (15.50 ± 2.08 versus 11.00 ± 2.35, 8.67 ± 3.21; p = 0.0037, p = 0.0183). IIEF-5 scores decreased significantly in patients with non-vascular ED postoperatively (15.50 ± 2.08 versus 10.00 ± 3.83; p = 0.0132), but were unchanged in patients with arterial/venous ED. CONCLUSION: Urethral trauma seriously affects erectile function, but subsequent end-to-end AU for urethral strictures has little impact.
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Disfunción Eréctil/etiología , Disfunción Eréctil/cirugía , Estrechez Uretral/cirugía , Procedimientos Quirúrgicos Urológicos/métodos , Adolescente , Adulto , Anastomosis Quirúrgica/efectos adversos , Anastomosis Quirúrgica/métodos , Estudios de Cohortes , Disfunción Eréctil/fisiopatología , Estudios de Seguimiento , Fracturas Óseas/complicaciones , Fracturas Óseas/diagnóstico por imagen , Humanos , Masculino , Persona de Mediana Edad , Pelvis/lesiones , Recuperación de la Función , Medición de Riesgo , Factores de Tiempo , Resultado del Tratamiento , Estrechez Uretral/diagnóstico por imagen , Estrechez Uretral/etiología , Urografía/métodos , Adulto JovenRESUMEN
This paper investigates the robust fault detection observer design problem in finite frequency domains for fractional-order singular systems. An H-/H∞ fault detection observer for fractional-order singular systems is designed to meet the system admissibility, disturbance robustness and fault sensitivity indices in finite frequency domains simultaneously. Four theorems and four corollaries about the system admissibility and the performance index H-/H∞ in different frequency ranges are given, and then the sufficient conditions are obtained in terms of linear matrix inequalities. Finally, two numerical examples are given to verify the validity of the presented methods.
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Protein phosphatase 2A (PP2A) complex comprises an extended family of intracellular protein serine/threonine phosphatases, that participate in different signaling transduction pathways. Different functions of PP2As are determined by the variety of regulatory subunits. In this study, CRISPR/Cas9-mediated loss-of-function screen revealed that PPP2R2A downregulation suppressed cell growth in NSCLC cells. AMOTL2 was identified and confirmed as a novel binding partner of PPP2R2A in NSCLC cells by mass spectrometry, CO-IP, GST pull-down and immunofluorescence. Upregulation of AMOTL2 also led to cell proliferation delay in human and mouse lung tumor cells. The proto-oncogene JUN is a key subunit of activator protein-1 (AP-1) transcription factor which plays crucial role in regulating tumorigenesis and its activity is negatively regulated by the phosphorylation at T239. Our results showed that either AMOTL2 upregulation or PPP2R2A downregulation led to great increase in JUN T239 phosphorylation. AMOTL2 bound PPP2R2A in cytoplasm, which reduced nuclear localization of PPP2R2A. In conclusion, AMOTL2 and PPP2R2A act respectively as negative and positive regulator of cell growth in NSCLC cells and function in the AMOTL2-PPP2R2A-JUN axis, in which AMOTL2 inhibits the entry of PPP2R2A into the nucleus to dephosphorylate JUN at T239.
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Carcinoma de Pulmón de Células no Pequeñas/genética , Proteínas Portadoras/genética , Proteína Fosfatasa 2/genética , Proteínas Proto-Oncogénicas c-jun/genética , Angiomotinas , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Sistema de Señalización de MAP Quinasas/genética , Fosforilación/genética , Proto-Oncogenes Mas , Factor de Transcripción AP-1/genética , Regulación hacia ArribaRESUMEN
Breathing is an integrated motor behavior that is driven and controlled by a network of brainstem neurons. Zfhx4 is a zinc finger transcription factor and our results showed that it was specifically expressed in several regions of the mouse brainstem. Mice lacking Zfhx4 died shortly after birth from an apparent inability to initiate respiration. We also found that the electrical rhythm of brainstemâspinal cord preparations was significantly depressed in Zfhx4-null mice compared to wild-type mice. Immunofluorescence staining revealed that Zfhx4 was coexpressed with Phox2b and Math1 in the brainstem and that Zfhx4 ablation greatly decreased the expression of these proteins, especially in the retrotrapezoid nucleus. Combined ChIPâseq and mRNA expression microarray analysis identified Phox2b as the direct downstream target gene of Zfhx4, and this finding was validated by ChIPâqPCR. Previous studies have reported that both Phox2b and Math1 play key roles in the development of the respiratory center, and Phox2b and Math1 knockout mice are neonatal lethal due to severe central apnea. On top of this, our study revealed that Zfhx4 is a critical regulator of Phox2b expression and essential for perinatal breathing.
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Apnea , Proteínas de Homeodominio/genética , Centro Respiratorio , Animales , Apnea/metabolismo , Apnea/mortalidad , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Encéfalo/metabolismo , Tronco Encefálico/metabolismo , Regulación de la Expresión Génica , Proteínas de Homeodominio/metabolismo , Ratones , Ratones Noqueados/genética , Neuronas/metabolismo , Respiración , Centro Respiratorio/embriología , Centro Respiratorio/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
BACKGROUND: Krüppel-like factor 16 (KLF16), a member of the KLF family, is involved in metabolism and regulation of the endocrine system and has emerging roles in tumor progression. However, the expression of KLF16 and its role in breast cancer are elusive. METHODS: We investigated the expression and prognostic value of KLFs in breast cancer using data acquired from the TCGA BRCA dataset and the Kaplan-Meier plotter dataset. The protein levels of KLF16 in breast specimens were detected by immunohistochemistry (IHC). KLF16 silencing using shRNAs was performed to explore the effects of KLF16 on breast cancer cell growth, migration, and invasion. The expression of EMT markers in cells manipulated for KLF16 expression was assessed by Western blotting. RESULTS: Using publicly available dataset and specimens from breast cancer patients, we found that the expression levels of KLF16 were significantly higher in tumor tissues and that high levels of KLF16 were associated with poor prognosis in breast cancer patients. Moreover, KLF16 expression levels had relation to several clinicopathological parameters of breast cancer, including the molecular subtype and histological grade. Importantly, knockdown of KLF16 dramatically suppressed cell proliferation both in vitro and in vivo. Also, KLF16 deletion impaired migration, and invasion in breast cancer cells, and suppressed epithelial-mesenchymal transition (EMT). CONCLUSION: Our results suggest that KLF16 has important oncogenic functions in breast cancer and that the expression levels of KLF16 are associated with prognosis in breast cancer patients. Our findings also suggest that KLF16 is involved in proliferation, migration, and invasion in breast cancer cells. Thus, KLF16 might be a promising prognostic marker and a therapeutic target for breast cancer.
RESUMEN
Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the GBA gene. Enzyme replacement treatment is the most effective therapy available for type 1 GD patients, but it is very expensive and does not improve neurologic outcomes in type 2 and 3 GD patients. This study evaluated the effectiveness of an adeno-associated virus 9 (AAV9) vector expressing the Gba gene delivered systemically in GD mouse models. To detect the therapeutic effects of the AAV9-mediated Gba transfer on the systemic symptoms of GD, an inducible whole-body Gba knockout mouse was developed in which tamoxifen effectively induced whole-body Gba gene deletion, and the mice displayed systemic symptoms of GD. The AAV9-CMV-Gba vector, with the expression of Gba driven by the universal CMV promoter, restored GCase activity in multiple organs and prolonged the lifespan in tamoxifen-induced GD mice after intravenous injection. Mice with brain-specific Gba deletion were also included in this study as a model of neuropathic GD (nGD) and injected intraperitoneally on postnatal day 5 with the AAV9-SYN-Gba vector; this improved the GCase activity, ameliorated the neuropathological changes and extended the mean lifespan two-fold. This study demonstrates that AAV9-mediated gene transfer is a potentially effective treatment for GD.
Asunto(s)
Dependovirus , Enfermedad de Gaucher , Terapia Genética , Vectores Genéticos , Glucosilceramidasa , Transducción Genética , Animales , Modelos Animales de Enfermedad , Enfermedad de Gaucher/enzimología , Enfermedad de Gaucher/genética , Enfermedad de Gaucher/patología , Enfermedad de Gaucher/terapia , Glucosilceramidasa/biosíntesis , Glucosilceramidasa/genética , Ratones , Ratones NoqueadosRESUMEN
Primary aldosteronism (PA) is the most common form of endocrine hypertension. This study was to investigate the gene expression profile in PA adrenal glands and normal controls using RNA-Sequencing. By performing transcriptome analyses for 3 PA adrenal glands and 3 controls on Illumina platform, we identified 1,093 transcripts as significantly differently expressed genes (DEGs), which provided clues for further study of these transcript changes during PA pathogenesis. Further, Gene Set Enrichment Analysis (GSEA) identified 35 significant Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathways, including 'ribosome', 'oxidative phosphorylation', 'histidine metabolism', 'xenobiotics metabolism by Cytochrome P450', 'drug metabolism by Cytochrome P450', 'tyrosine metabolism' and 'glutathione metabolism'. In summary, we identified novel genes that are associated with PA phenotype, as well as differently regulated biological pathways relating to protein synthesis, energy acquisition and metabolism. Our study provides new candidates for further elucidation of the molecular mechanisms underlying PA pathogenesis.