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OBJECTIVE: To study the prevalence, epidemiological characteristics, and risk factors for childhood asthma in Yichang City, China and to collect evidence for the early diagnosis and preventive treatment of asthma. METHODS: Preliminary screening questionnaires were distributed to more than 90% of children in 5 kindergartens, 10 primary and secondary schools, and 5 communities in Yichang City to detect children with suspected asthma. These surveyed children were selected by cluster random sampling. A further questionnaire survey was conducted for suspected cases. Meanwhile, a similar number of sex- and age-matched non-asthmatic children were selected for the case-control study. Information from returned questionnaires was entered into a database for statistical analysis. RESULTS: A total of 11 000 questionnaires were distributed, and 10 456 (95.1%) questionnaires were returned. The prevalence rate of asthma among children in Yichang was 3.47%, significantly higher in boys than in girls (P<0.05). A total of 107 out of 363 children with asthma had a history of drug allergy, and 152 cases had a family history of allergy. The majority of asthmatic children had irregular onset-prone seasons and hours. Respiratory tract infections were the most common trigger of asthma attacks, accounting for 93.1% of all onsets; family history of allergy, history of early use of antibiotics, history of housing renovation, and history of passive smoking were the major risk factors for asthma. CONCLUSIONS: Prevention of respiratory tract infections may reduce the frequency of asthma attacks; reducing the use of antibiotics during early childhood, decreasing the frequency of housing renovation, and advocating for smoking cessation among parents have preventive effects on asthma.
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Asma/epidemiología , Adolescente , Asma/etiología , Asma/prevención & control , Índice de Masa Corporal , Niño , Preescolar , China/epidemiología , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Prevalencia , Infecciones del Sistema Respiratorio/complicaciones , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
The present study reports a patient case with a 17α-hydroxylase deficiency accompanied by triple X syndrome. A 17α-hydroxylase deficiency leads to a very low 17α-hydroxylated steroid synthesis as well as a non-feedback increase in the adrenocorticotropic hormone level. Meanwhile, the progesterone level increases the 17α-hydroxyprogesterone level and decreases the dehydroepiandrosterone sulfate level. The patient is characterized by intractable hypokalemia, high urinary potassium, hyperaldosteronemia, hyporeninemia, hypocortisolemia, hypertension, gonadal and secondary sexual dysplasia, a decreased estrogen level, primary amenorrhea, and infertility. The imaging findings indicate a presence of multiple bilateral adrenal gland adenomas, and the sequencing indicates a missense CYP17A1-E7 gene pathogenic variant. The karyotype is a 47, XXX [3]/46, XX [47] low-level chimeric karyotype. The patient's parents are cousins. To our knowledge, this patient is the first case diagnosed with congenital adrenal hyperplasia caused by hydroxylase deficiency and triple X syndrome. The uniqueness of this case is that this patient has two very rare genetic diseases, probably due to the marriage of close relatives.
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OBJECTIVE: To investigate the effects of methionine restriction on proliferation, cell cycle and apoptosis of human acute leukemia cells. METHODS: Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of methionine restriction on HL-60 and Jurkat cells proliferation. The effect of methionine restriction on cell cycle of HL-60 and Jurkat cells was examined by PI staining. Annexin V-FITC / PI double staining was applied to detect apoptosis of HL-60 and Jurkat cells following methionine restriction. The expression of cell cycle-related proteins cyclin B1, CDC2 and apoptosis-related protein Bcl-2 was evaluated by Western blot assay. RESULTS: Methionine restriction significantly inhibited the proliferation of HL-60 and Jurkat cells in a time-dependent manner (HL-60: r =0.7773, Jurkat: r =0.8725), arrested the cells at G2/M phase (P < 0.001), and significantly induced apoptosis of HL-60 and Jurkat cells (HL-60: P < 0.001; Jurkat: P < 0.05). Furthermore, Western blot analysis demonstrated that methionine restriction significantly reduced the proteins expression of Cyclin B1 (P < 0.05), CDC2 (P < 0.01) and Bcl-2 (P < 0.001) in HL-60 and Jurkat cells. CONCLUSION: Acute leukemia cells HL-60 and Jurkat exhibit methionine dependence. Methionine restriction can significantly inhibit the proliferation, promote cell cycle arrest and induce apoptosis of HL-60 and Jurkat cells, which suggests that methionine restriction may be a potential therapeutic strategy for acute leukemia.
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Leucemia Mieloide Aguda , Metionina , Humanos , Ciclina B1/genética , Ciclina B1/metabolismo , Ciclina B1/farmacología , Proliferación Celular , Metionina/farmacología , Ciclo Celular , Apoptosis , División Celular , Proteínas de Ciclo Celular , Células Jurkat , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Células HL-60RESUMEN
OBJECTIVE: To investigate the inhibitory effects of novel phosphodiesterase 4 inhibitor ZL-n-91 to the proliferation of leukemia cells L1210 and K562. METHODS: CCK-8 method was used to detect the effect of ZL-n-91 to the proliferation of L1210 and K562 cells, and the proliferation rate, IC50 were calculated. The effects of ZL-n-91 to the cycle of L1210 and K562 cells was detected by PE single staining, and the effects of ZL-n-91 to the apoptosis of L1210 and K562 cells was detected by PE/7AA-D double staining. Western blot was used to detect the effect of ZL-n-91 to the expression levels of apoptosis related proteins. Subcutaneous tumor transplantation model of acute lymphoblastic leukemia L1210 was established in the nude mice, and the inhibitory effect of oral administration of ZL-n-91 to the xenograft was observed. RESULTS: ZL-n-91 showed a significant inhibitory effect to the proliferation of leukemia cells L1210 and K562 in a dose-dependent manner (P<0.001). After treated by ZL-n-91, the leukemia cells L1210 and K562 in the S-phase in cell cycle decreased significantly compared with those in control group (P<0.01). The apoptosis of leukemia cells L1210 and K562 could be induced by ZL-n-91 (P<0.001), and the expression level of apoptosis related protein BAX significantly increased. In the animal experiment, the result showed that ZL-n-91 could significantly inhibit the growth of subcutaneously transplantation tumor (P<0.05). CONCLUSION: The novel phosphodiesterase 4 inhibitor ZL-n-91 can effectively inhibit the proliferation of leukemia cells L1210 and K562, which has the potential of anti-leukemia drug development.
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Leucemia , Inhibidores de Fosfodiesterasa 4 , Animales , Proliferación Celular , Humanos , Células K562 , Ratones , Ratones Desnudos , Inhibidores de Fosfodiesterasa 4/farmacologíaRESUMEN
To determine the prevalence and clinical features of olfactory and taste disorders among coronavirus disease 2019 (COVID-19) patients in China. A cross-sectional study was performed in Wuhan from April 3, 2020 to April 15, 2020. A total of 187 patients with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) completed face-to-face interviews or telephone follow-ups. We found that the prevalence of olfactory and taste disorders was significantly lower in the Chinese cohort than in foreign COVID-19 cohorts. Females were more prone to olfactory and taste disorders. In some patients, olfactory and taste disorders precede other symptoms and can be used as early screening and warning signs.
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COVID-19/complicaciones , Trastornos del Olfato/etiología , Olfato , Trastornos del Gusto/etiología , Gusto , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , China/epidemiología , Estudios Transversales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Trastornos del Olfato/epidemiología , Prevalencia , SARS-CoV-2 , Factores Sexuales , Trastornos del Gusto/epidemiología , Adulto JovenAsunto(s)
Prepucio/citología , Prepucio/crecimiento & desarrollo , Folículo Piloso/citología , Folículo Piloso/crecimiento & desarrollo , Cabello/citología , Cabello/crecimiento & desarrollo , Regeneración/fisiología , Adulto , Pueblo Asiatico , Células Cultivadas/fisiología , Niño , Humanos , MasculinoRESUMEN
OBJECTIVE: To study the spectrum of mutations in methyl-CpG-binding protein 2 gene (MECP2) and cyclin-dependent kinase-like 5 gene (CDKL5) in Chinese pediatric patients with Rett syndrome (RTT), and establish a simple, quick, and efficient gene test method as well as screen a strategy of genetic diagnosis for RTT. METHODS: Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes of 117 pediatric patients diagnosed from 1987 to 2007. PCR was used to amplify the exons 1 - 4 of MECP2 using published primers. If no mutation was identified after screening exons 2 - 4, exon 1 was screened. If no mutation was identified in MECP2 by sequencing, multiplex ligation dependent probe amplification (MLPA) was employed to screen for large deletions by using P015C kit. If no mutation was identified in the MECP2 by sequencing and MLPA respectively, then the coding region of CDKL5 was screened by denaturing high performance liquid chromatography (DHPLC). RESULTS: The total mutation frequency in MECP2 and CDKL5 genes among all RTT patients was 82%. MECP2 mutations were found in 86% (137/159) of the patients with classical RTT and in 44% (8/18) of those with atypical RTT. Most of the mutations were missense mutations, accounting for 39%, followed in order of frequency by nonsense mutations 28%, frame shift mutations 17% and large deletions 14.5%. The eight most frequent MECP2 mutations were p.T158M (13%), p.R168X (12%), c.806delG (7%), p.R255X (6%), p.R270X (5%), p.R133C (5%), p.R306C (4%), and p.R106W (3%), with p.T158M as the most common of the MECP2 mutations and c.806delG as a hotspot mutation in Chinese patients with RTT. Only one synonymous mutation was identified in CDKL5. CONCLUSION: The spectrum of MECP2 mutations within the mainland Chinese RTT patients is similar to that of those patients reported in the world. p.T158M, p.R168X, c.806delG, p.R255X, p.R270X, p.R133C, p.R306C, and p.R106W are the hotspot mutations of MECP2 and c.806delG is a specific hotspot mutation in Chinese patients with RTT. The most effective method to screen mutations is to screen the exon 4. MLPA is an effective supplement to the routine methods.
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Proteína 2 de Unión a Metil-CpG/genética , Mutación , Proteínas Serina-Treonina Quinasas/genética , Síndrome de Rett/genética , Pueblo Asiatico/genética , Preescolar , Análisis Mutacional de ADN , Exones , Genotipo , Humanos , LactanteRESUMEN
OBJECTIVE: To study the rate of subtelomeric rearrangements in patients with idiopathic mental retardation (MR) and to search the cause of MR. METHODS: DNA was extracted and purified from peripheral blood leukocytes of 180 patients with idiopathic MR. DNA was tested using specific subtelomeric multiplex ligation-dependent probe amplification (MLPA) kits P036B/C and P070 according to manufacturer's instructions. The amplification products were separated by capillary electrophoresis using an ABI 3100 automated sequencer and size standard. MLPA data were extracted by GeneScan Analysis software. Data normalization and analysis were performed with the built-in MLPA application in GeneMarker. RESULTS: Among 180 patients with idiopathic MR, 12 had pathological subtelomeric deletions including 3 cases with a 4p deletion, 2 cases with a deletion at 9q and 22q respectively, 1 case with a deletion at 1p, 7p, 8p, 9q and 12q respectively. Subtelomeric rearrangements were responsible for 7% cases of idiopathic mental retardation. CONCLUSION: Subtelomeric rearrangement is a common cause of idiopathic mental retardation.
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Reordenamiento Génico , Discapacidad Intelectual/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Telómero/genética , Adolescente , Niño , Preescolar , Cromosomas , Femenino , Humanos , Lactante , Cariotipificación , MasculinoRESUMEN
OBJECTIVE: Currently, no agent has been conclusively demonstrated to prevent the progression of non-alcoholic steatohepatitis (NASH). Chitosan, a natural product derived from chitin, was thought to possess hypocholesterolemic properties. The aim of this study was to evaluate the potential effects of chitosan on nutritional steatohepatitis in rats. MATERIAL AND METHODS: Rats were fed with a high fat diet for 4 weeks to develop NASH that was confirmed by liver biopsy, and then 4 weeks of chitosan was given. Serum chemistry and liver histology were assessed and the steatoinflammatory mechanisms were studied. RESULTS: Chitosan significantly protected against high fat diet-induced hepatic steatohepatitis. This effect was associated with repressed serum levels of total protein (TP), globulin (GLO), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), total cholesterol (TC) and low-density lipoprotein (LDL). Chitosan elevated the serum levels of high-density lipoprotein (HDL) and the ratio of albumin to globulin. Furthermore, increased TNF-alpha, lipoemia, hyperinsulinemia, hyperleptinemia and hypoadiponectin in NASH were significantly ameliorated by treatment with chitosan. CONCLUSIONS: Chitosan effectively attenuated the steatohepatitis induced by a high fat diet. The therapeutic effect of chitosan on NASH may be activated through exerting an influence on adipokines.
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Anticolesterolemiantes/uso terapéutico , Quitosano/uso terapéutico , Grasas de la Dieta/efectos adversos , Hígado Graso/tratamiento farmacológico , Animales , Modelos Animales de Enfermedad , Hígado Graso/etiología , Masculino , Ratas , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
OBJECTIVE: To verify the sensitivity and reliability of methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) and to develop a simple, accurate, reliability method of genetic diagnosis for AS and PWS. METHODS: Peripheral blood samples were collected from 4 suspected AS patients, 2 suspected PWS patients, 2 normal persons, and 2 molecular biologically proven positive controls (1 AS patient and 1 PWS patient). DNA was extracted and purified. MS-MLPA was used to detect the methylation of the CpG dinucleotide and the copy number in the 15q-q13 region. The results of MS-MLPA were confirmed by MSP. RESULTS: Three cases with maternal deletion on 15q11-q13 region and one case with paternal uniparental disomy (UPD) or imprinting center defect in 15q11-q13 region were found in the 4 suspected AS patients. One PWS case was found to be with paternal deletion in 15q11-q13 region and the other with paternal deletion in 15q11-q13 region or UPD or imprinting center defect in 15q11-q13 region. CONCLUSION: MS-MLPA is a simple, rapid, accurate, and reliable method of genetic test.
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Síndrome de Angelman/diagnóstico , Técnicas de Amplificación de Ácido Nucleico/métodos , Síndrome de Prader-Willi/diagnóstico , Síndrome de Angelman/genética , Niño , Cromosomas Humanos Par 15/genética , Sondas de ADN , Humanos , Metilación , Síndrome de Prader-Willi/genética , Juego de Reactivos para DiagnósticoRESUMEN
Traditional Chinese medicine has great potential to improve wound healing. ANBP, the mixture of 4 Chinese herbs- Agrimoniapilosa, Nelumbonucifera, Boswelliacarteri, and Pollen typhae-is effective in trauma treatment while its mechanism is still elusive. In this study, quantitative proteomics and bioinformatics analyses were performed to decipher the possible roles of ANBP in accelerated wound healing of mouse skin. Among all 3171 identified proteins, 90, 71, 80, and 140 proteins were found to be differently expressed in 6 hours, 3 days, 7 days, and 14 days ANBP-treated tissues compared with corresponding control tissues, respectively. The result showed that different biological processes and pathways were activated at different healing stages. At the early healing stage, ANBP treatment mainly affected several biological processes, including immune and defense response, vascular system restoration, hemostasis and coagulation regulation, lipid metabolism and signal transduction, while muscle tissue, hair, epidermis, extracellular matrix and tissue remodeling related activities were the major events in ANBP promoted later wound healing. This is the first quantitative proteome study of ANBP-treated wound tissues, which provide a new perspective for the mechanism of ANBP accelerated wound healing and is of guiding significance for clinical application of ANBP in trauma disorders cure.
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Medicamentos Herbarios Chinos/uso terapéutico , Proteómica , Cicatrización de Heridas/efectos de los fármacos , Heridas y Lesiones/tratamiento farmacológico , Heridas y Lesiones/patología , Animales , Biopsia con Aguja , Modelos Animales de Enfermedad , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos C57BL , Distribución Aleatoria , Valores de Referencia , Sensibilidad y Especificidad , Piel/efectos de los fármacos , Piel/patología , Cicatrización de Heridas/genética , Heridas y Lesiones/genéticaRESUMEN
OBJECTIVE: To find the valid siRNA (small interference RNA) sequence to knock down rat Mecp2 expression for the analysis of Mecp2 function by RNA interference (RNAi). METHODS: A plasmid (pMecp2-RFP) expressing rat Mecp2 and marker gene red fluorescent protein (RFP) as a fusion gene was constracted. We first selected a candidate valid sequence by cotransfecting the pMecp2-RFP with 4 candidate siRNAs targeting rat Mecp2 to HEK293T cells respectively, and then identified the RNAi efficiency of the candidate valid siRNA by detecting its effect on suppressing the expression of endogenous rat Mecp2 in PC12 cells. RESULTS: siRNA sequence (5'-GCUGUGAAGGAAUCUUCUA-3') targeting rat Mecp2 mRNA 918-936nt was the valid sequence to knock down rat Mecp2 expression by RNA interference. As the target sequence was located in exon 4 of rat Mecp2 mRNA, we assumed that it could suppress the expression of Mecp2alpha and Mecp2beta at the same time. And as the target sequence was located in the coding region of rat Mecp2, we assumed that it could suppress the expression of both 1.9 kb and 10 kb of rat Mecp2 transcript. CONCLUSION: This study has laid the foundation for constructing rat Mecp2 knocked-down neuronal cell model to study the gene function of Mecp2 in brain development.
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Proteína 2 de Unión a Metil-CpG/genética , Interferencia de ARN , ARN Interferente Pequeño/genética , Animales , Encéfalo/citología , Encéfalo/metabolismo , Línea Celular , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteína 2 de Unión a Metil-CpG/metabolismo , Microscopía Fluorescente , Células PC12 , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Transfección , Proteína Fluorescente RojaRESUMEN
BACKGROUND: Trichophyton rubrum is superficial fungi characteristically confined to dead keratinized tissues. These observations suggest that the soluble components released by the fungus could influence the host immune response in a cell in contact-free manner. Therefore, this research aimed to analyze whether the culture supernatant derived from T. rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively. METHODS: The culture supernatants of two strains (T1a, T XHB ) were compared for the ß-glucan concentrations and their capacity to impact the innate immunity of keratinocytes. The ß-glucan concentrations in the supernatants were determined with the fungal G-test kit and protein concentrations with bicinchoninic acid protein quantitative method, then HaCaT was stimulated with different concentrations of culture supernatants by adopting morphological method to select a suitable dosage. Expressions of host defense genes were assessed by quantitative polymerase chain reaction after the HaCaT was stimulated with the culture supernatants. Data were analyzed with one-way analysis of variance, followed by the least significant difference test. RESULTS: The T. rubrum strains (T1a and T XHB ) released ß-glucan of 87.530 ± 37.581 pg/ml and 15.747 ± 6.453 pg/ml, respectively into the media. The messenger RNA (mRNA) expressions of toll-like receptor-2 (TLR2), TLR4, and CARD9 were moderately up-regulated in HaCaT within 6-h applications of both supernatants. HaCaT cells were more responsive to T1a than T XHB . The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant than T XHB . The moderate decreases of RNase 7, the slight up-regulations of Dectin-1 and interleukin-8 at the mRNA level were detected only in response to T1a rather than T XHB . After a long-time contact, all the elevated defense genes decreased after 24 h. CONCLUSION: The culture supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.
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Medios de Cultivo Condicionados/farmacología , Inmunidad Innata/efectos de los fármacos , Queratinocitos/efectos de los fármacos , Queratinocitos/metabolismo , Trichophyton/metabolismo , Línea Celular Tumoral , Humanos , beta-Glucanos/metabolismoRESUMEN
Wound healing is a troublesome problem in diabetic patients. Besides, there is also an increased risk of postsurgical wound complications for diabetic patient. It has been revealed that traditional Chinese medicine may promote healing and inhibit scar formation, while the changes of morphology and physiology of wounds on such medicine treatment still remain elusive. In this study, we first used the ultralow temperature preparation method to produce mixed superfine powder from Agrimonia pilosa (A), Nelumbo nucifera (N), Boswellia carteri (B), and Pollen typhae (P), named as ANBP. Applying ANBP on 40 streptozotocin (STZ)-induced diabetic C57BL/6 mice (4-6 weeks, 20 ± 2 g), we observed that the wound healing process was accelerated and the wound healing time was shortened (14 days, P < .05). Pathological observation using hematoxylin-eosin staining indicated that inflammatory cells were reduced (P < .05) while the thickness of granulation tissue and length of epithelial tongue were increased (P < .05). The vascular density was increased on 7 and 14 days after ANBP treatment. Masson and Sirius red staining showed that, at the early stage of trauma, the expressions of Col I and Col III, especially Col III, were increased in the ANBP group (P < .05). Studies in vitro demonstrated that tubular formation was significantly increased after ANBP treatment on human vascular endothelial cells in a dose-dependent way. Taken together, our studies revealed that ANBP treatment could accelerate wound healing, promote vascularization, and inhibit inflammation, suggesting the potential clinic application of ANBP for diabetes mellitus and refractory wounds.
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Diabetes Mellitus Experimental , Medicamentos Herbarios Chinos/uso terapéutico , Fitoterapia , Cicatrización de Heridas/efectos de los fármacos , Animales , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
BACKGROUND: Trichophyton rubrum (T. rubrum) represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that it may result from the failure of epithelial cells to recognize T. rubrum effectively and initiate effective immune responses. The C-type lectin receptors (CLR) and toll-like receptors (TLR) are the two major pattern recognition receptors (PRRs) that recognize fungal components. Therefore, the purpose of the study was to analyze the expression of those PRRs and the cytokines in HaCaT cells stimulated with heat-inactivated T. rubrum conidia and hyphae, respectively. METHODS: HaCaT cells were unstimulated or stimulated with heat-inactivated T. rubrum conidia and hyphae (1×10(6) and 1.5×10(5) colony-forming unit (CFU) in 2 ml medium, respectively) for 6, 12 and 24 hours. The mRNA expression of PRRs involved in recognizing fungal pathogen-associated molecular patterns (PAMPs) and signaling molecules were measured by quantitative reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, surface toll-like receptor (TLR) 2, TLR4 and Dectin-1 were analyzed by fluorescence-activated cell sorter (FACS) 24 hours after treatment. The cytokines were detected in cell culture supernatants of HaCaT cells in 12 and 24 hours after treatment. RESULTS: HaCaT cells constitutively expressed mRNA of membrane-bound TLR1, 2, 4 and 6, Dectin1 and DC-SIGN, but not Dectin-2 or Mincle. Heat-killed T. rubrum did not significantly upregulate gene transcriptions of the PRRs of HaCaT cells. Heat-inactivated T. rubrum conidia significantly reduced the surface expression of TLR2 and Dectin-1, and suppressed the secretions of interferon-inducible protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) of HaCaT cells, while heat-killed T. rubrum hyphae significantly induced the secretions of IP-10 and MCP-1. CONCLUSION: The cell-wall antigens of T. rubrum fail to activate transcriptional expression of PRRs and induce a lower immune response of HaCaT cells by limited cytokines secretion.
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Queratinocitos/inmunología , Trichophyton/inmunología , Células Cultivadas , Citocinas/biosíntesis , Humanos , Lectinas Tipo C/genética , Lectinas Tipo C/fisiología , ARN Mensajero/análisis , Receptores de Reconocimiento de Patrones/genética , Receptores de Reconocimiento de Patrones/fisiología , Receptor Toll-Like 2/fisiologíaRESUMEN
Seeking possible ways to create replacement cells for the faded ones with deficits in functionality or quantity inspires comprehensive needs for cell lineage conversion. To fulfill this promise, reprogramming and microenvironment direction have been used to manipulate abundant cell fates. We briefly describe the evolution and fundamental insights of these two major strategies applied for lineage specification, comment generally on their current limitations, and analyze the orchestral interplay between them. We also present several future directions and discuss the potential clinical uses. Based on the relatively slight safety and technical issues, we conclude that microenvironment-evoked cell lineage conversion, instead of reprogramming, will be the shifting focus in regenerative medicine.
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Linaje de la Célula/fisiología , Ambiente , Envejecimiento/fisiología , Animales , Reprogramación Celular/genética , Reprogramación Celular/fisiología , Técnicas de Cocultivo , Técnicas Citológicas , Humanos , Células Madre Pluripotentes Inducidas , Células Madre Pluripotentes/fisiologíaRESUMEN
BACKGROUND: Diabetic cardiomyopathy is the major cause of morbidity and mortality in diabetic patients. Oxidative stress plays an important role in diabetic cardiomyopathy. This study aimed to investigate the effects of adiponectin on oxidative stress and apoptosis in human cardiac myocytes (HCM) cultured with high glucose. METHODS: The cells were assigned to three group: control group, high glucose group and high glucose plus adiponectin group. After culture for 24, 48, 72 hours, oxidative stress was evaluated by detecting levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the supernatant of culture media. The expression of p66Shc and Heme oxygenase-1 (HO-1) was detected by real-time polymerase chain reaction (PCR). Flow cytometry was designed to observe and detect cellular apoptosis. RESULTS: Our findings showed significant increase in MDA levels and decrease in SOD activity in the high glucose group compared with the control group (P < 0.05). However, MDA levels were significantly decreased and SOD activity was significantly increased in the adiponectin group compared with those in the high-glucose group (P < 0.05). The mRNA expression of HO-1 in the high glucose group was significantly increased in a time-dependent manner compared with that in the control group (P < 0.05). Adiponectin further increased the mRNA expression of HO-1 induced by high glucose in a time-dependent manner (P < 0.05).The expression of p66Shc was significantly increased in high glucose group compared with that in the control group (P < 0.05). Adiponectin significantly suppressed the upregulation of p66Shc induced by high glucose (P < 0.05). The apoptotic rate of cardiomyocytes was significantly increased in the high glucose group compared with that in the control group while the apoptotic rate in the adiponectin group was remarkably declined in comparison with that in the high glucose group. CONCLUSION: Adiponectin reduces high glucose-induced oxidative stress and apoptosis and plays a protective role in myocardial cells by upregulating the HO-1 expression and downregulating p66Shc expression.
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Adiponectina/farmacología , Apoptosis/efectos de los fármacos , Glucosa/farmacología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Línea Celular , Citometría de Flujo , Humanos , Malondialdehído/metabolismo , Miocitos Cardíacos/citología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Superóxido Dismutasa/metabolismoRESUMEN
OBJECTIVE: To investigate the effects of Trichophyton rubrum exposure on the expressions of toll-like receptor-2 (TLR-2), TLR-4 and dendritic cell associated C-type lectin-1 (Dectin-1) and cytokine secretions in human keratinocytes cell line HaCaT. METHODS: The mRNA of TLR-2,4, and dectin-1 in the HaCaT co-cultured with the conidia of Trichophyton rubrum conidia for 24 h was measured with real-time PCR. The mean fluorescence intensity and the percentage of cells positive for TLR-2, 4, and dectin-1 was detected during the co-culture using flow cytometry. The cytokine secretion profiles in the cell culture supernatant was analyzed using a cytokine antibody array. RESULTS: The TLR-2,4, and dectin-1 mRNA expressions, mean fluorescence intensity and percentage of positive cells for TLR-2,4, and dectin-1 all increased in HaCaT cells in response to Trichophyton rubrum conidia exposure. The results of cytokine antibody array demonstrated obviously increased secretions of IL-8, I-309, IFN-γ, IL-6, and IL-13 in the culture supernatant of HaCaT cells in response to Trichophyton rubrum exposure. CONCLUSION: The immune responses and immunological recognition of human keratinocytes to Trichophyton rubrum conidia are partially mediated by up-regulating the expressions of TLR-2, TLR-4 and dectin-1 and secretions of multiple cytokines.
Asunto(s)
Queratinocitos/metabolismo , Lectinas Tipo C/metabolismo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo , Trichophyton , Línea Celular , Quimiocina CCL1/metabolismo , Técnicas de Cocultivo , Humanos , Interferón gamma/metabolismo , Interleucina-13/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE: Rett syndrome (RTT) is a neurodevelopmental disorder occurring almost exclusively in females as sporadic cases due to de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Familial cases of RTT are rare and are due to X-chromosomal inheritance from a carrier mother. Recently, DNA mutations in the MECP2 have been detected in approximately 84.7% of patients with RTT in China. To explain the sex-limited expression of RTT, it has been suggested that de novo X-linked mutations occur exclusively in male germ cells resulting therefore only in affected daughters. To test this hypothesis, we have analyzed the parental origin of mutations and the XCI status in 15 sporadic cases with RTT due to MECP2 molecular defects. METHODS: Allele-specific PCR was performed to amplify a fragment including the position of the mutation. The allele-specific PCR products were sequenced to determine which haplotype contained the mutation. It was then possible to determine the parent of origin by genotyping the single nucleotide polymorphism (SNP) in the parents. The degree of XCI and its direction relative to the X chromosome parent of origin were measured in DNA prepared from peripheral blood leucocytes by analyzing CAG repeat polymorphism in the androgen receptor gene (AR). RESULTS: Except for 2 cases who had a frameshift mutation; all the remaining 13 cases had a C-->T transition mutation. Paternal origin has been determined in all cases with the C-->T transition mutation. For the two frameshift mutations, paternal origin has been determined in one case and maternal origin in the other. The frequency of male germ-line transmission in mutations is 93.3%. Except for 2 cases who were homozygotic at the AR locus, of the remaining 13 cases, 8 cases had a random XCI pattern; the other five cases had a skewed XCI pattern and they favor expression of the maternal origin allele. CONCLUSION: De novo mutations in sporadic RTT occur almost exclusively on the paternally derived X chromosome and that this is most probably the cause for the high female: male ratio observed in sporadic cases with RTT. Random XCI was the main XCI pattern in sporadic RTT patients. The priority inactive X chromosome was mainly of paternal origin.
Asunto(s)
Proteína 2 de Unión a Metil-CpG/genética , Síndrome de Rett/genética , Inactivación del Cromosoma X , Aberraciones Cromosómicas , Cromosomas Humanos X , Femenino , Humanos , Masculino , Mutación , Polimorfismo de Nucleótido SimpleRESUMEN
OBJECTIVE: Rett syndrome (RTT) is a neurodevelopmental disorder that represents one of the most common genetic causes of mental retardation in girls. The aim of this study was to investigate the correlation between MECP2 genotype and phenotype and thereby not only to provide assistance for clinical care, but also facilitate clinical genetic counseling. METHOD: Individual phenotype characteristic and clinical severity of 126 children with RTT diagnosed by molecular genetic methods were evaluated by using scales of Kerr et al and Scala et al. Statistical package SPSS 12.0 was used for analyses of data. Since the majority of the data were not normally distributed, non-parametric tests were used. The Kruskal-Wallis test/Wilcoxon Mann-Whitney test was employed to compare total severity phenotype scores. The Fisher exact test was used for comparing rates. Statistical significance was set at P < 0.05. RESULT: There were no significant differences in the average overall scores for RTT patients with mutations in the region of methyl-CpG-binding domain (MBD) compared with those mutations in the transcription repression domain (TRD) and C terminal segment (CTS), also patients with nonsense mutations compared with missense mutations, frameshift mutations and large deletions (P > 0.05). The RTT patients with nonsense mutations located in the region of MBD have more severe phenotype than those with missense mutations in the same region (P = 0.016). Among p.T158M, p.R168X, c.806delG and p.R255X, there were no significant differences in the average overall scores (P > 0.05), but there were significant differences in language skill (P = 0.028) and in language impairment rate at different level (P = 0.019). CONCLUSION: There are relationships between MECP2 genotype and phenotype:the RTT patients with nonsense mutations located in MBD tend to develop more severe phenotype;there are significant differences in language skill and language impairment rate in the groups with p.T158M, p.R168X, c.806del and p.R255X, which had higher frequency in children below five-years of age and the p.R168X present with most severe impairment.