RESUMEN

Juvenile in vitro embryo transfer is an important animal reproductive technology that can shorten the generation interval of livestock, explore the reproductive potential of dams with excellent genetic traits, accelerate genetic progress and production efficiency of the herd, and provide a wealth of genetic resources for livestock breeding. However, oocytes from kids do not develop as well as those from female goats during in vitro maturation. To identify differences during different stages of oocyte maturation, we used single cell transcriptome sequencing to compare gene expression in mature oocytes from kids and female goats. We identified 1086 differentially expressed genes in mature oocytes from kids and female goats. Of these, we observed upregulated expression in 355 genes and downregulated expression in 435 genes. The differentially expressed genes were involved in a total of 245 different pathways; of which 30 were significant (P ≤ 0.05). We used real-time quantitative polymerase chain reaction to screen and verify the expression of five genes specifically involved in oocyte maturation (MOS, RPS6KA1, CPEB1, ANAPC13, and CDK1). Further study of these genes will be of great importance for improving the reproductive performance of Haimen white goats.

Asunto(s)

Cabras/genética , Oocitos/fisiología , Transcriptoma , Animales , Cruzamiento , Células Cultivadas , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Cabras/metabolismo , Oogénesis , Análisis de la Célula Individual

RESUMEN

The aims of the current study were to assess the clinical features of allergic rhinitis (AR) in children in Shanghai. Serum-specific IgE (sIgE) tests were performed on samples from patients with AR symptoms from January 2011 to December 2014. A disease-related questionnaire was completed after AR diagnosis. The allergen profile and clinical features of AR were analyzed. In total, 2713 AR patients were enrolled in this study. Dermatophagoides pteronyssinus was found to be the most common offending allergen in the study population. With increasing age, the prevalence of sIgE against inhalant allergens was significantly increased; however, the opposite trend was observed for food allergens. Additionally, the proportion of children with high levels of sIgE against D. pteronyssinus increased with age. Of the AR cases, 8.6% were classified as intermittent mild, 4.2% as persistent mild, 40.5% as intermittent moderate-severe, and 46.7% as persistent moderate-severe. A family history of allergies and a patient history of allergies within 6 months of birth were significantly associated with the duration and severity of AR symptoms. The occurrence of co-morbidities, such as allergic conjunctivitis, cough, and asthma, gradually increased from intermittent mild, persistent mild, and intermittent moderate-severe to persistent moderate-severe. The most frequently used drugs were topical corticosteroids and oral antihistamines, which were used by 86.7 and 79.0% of patients, respectively. These results confirm the adequacy of the Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines for classifying AR patients, and advance the understanding of clinical features of AR in children in Shanghai, China.

Asunto(s)

Rinitis Alérgica/inmunología , Adolescente , Alérgenos/inmunología , Animales , Niño , Preescolar , China/epidemiología , Dermatophagoides pteronyssinus/inmunología , Femenino , Humanos , Inmunoglobulina E/sangre , Masculino , Prevalencia , Estudios Prospectivos , Rinitis Alérgica/sangre , Rinitis Alérgica/epidemiología , Rinitis Alérgica/etiología , Encuestas y Cuestionarios

RESUMEN

The objective of this study was the development of a gene/protein interaction network for primary myelofibrosis based on gene expression, and the enrichment analysis of KEGG pathways underlying the molecular complexes in this network. To achieve this, genes involved in primary myelofibrosis were selected from the OMIM database. A gene/protein interaction network for primary myelofibrosis was obtained through Cytoscape with the literature mining performed using the Agilent Literature Search plugin. The molecular complexes in the network were detected by ClusterViz plugin and KEGG pathway enrichment of molecular complexes was performed using DAVID online. We found 75 genes associated with primary myelofibrosis in the OMIM database. The gene/protein interaction network of primary myelofibrosis contained 608 nodes, 2086 edges, and 4 molecular complexes with a correlation integral value greater than 4. Molecular complexes involved in KEGG pathways are related to cytokine regulation, immune function regulation, ECM-receptor interaction, focal adhesion, actin cytoskeleton regulation, cell adhesion molecules, and other biological behavior of tumors, which can provide a reliable direction for the treatment of primary myelofibrosis and the bioinformatic foundation for further understanding the molecular mechanisms of this disease.

Asunto(s)

Regulación de la Expresión Génica , Redes Reguladoras de Genes , Mielofibrosis Primaria/genética , Mielofibrosis Primaria/metabolismo , Mapas de Interacción de Proteínas , Transducción de Señal , Análisis por Conglomerados , Biología Computacional/métodos , Bases de Datos Genéticas , Humanos

RESUMEN

CD14 is involved in primary immune and inflammatory responses. The -159 C/T variation in the CD14 gene plays an important role in regulating CD14 expression and has been associated with the susceptibility to various diseases, including allergies. In this study, we examined the association between the C-159T polymorphism and atopic asthma susceptibility in children from Southeastern China. The study population included 746 unrelated children of Chinese Han nationality (362 patients with atopic asthma and 384 healthy controls). CD14 gene polymorphisms were identified by direct sequencing of polymerase chain reaction products. Total immunoglobulin E (IgE) levels in human serum samples were determined using an enzyme-linked immunosorbent assay. Individuals carrying the TT genotypes for rs2569190 were significantly associated with an increased risk of atopic asthma compared with those carrying the wild-type homozygous CC genotypes [adjusted odds ratio (OR) by gender and age, from 1.075-2.398, P = 0.025]. Total serum IgE levels in TT genotype carriers were significantly higher than those in CC genotype carriers in atopic asthma patients (286.3 ± 161.5 IU/mL vs 248.3 ± 147.8 IU/mL). Our data suggest that the CD14 TT genotype may be a genetic susceptibility marker for atopic asthma in Chinese Han children.

Asunto(s)

Asma/genética , Frecuencia de los Genes/genética , Inmunoglobulina E/sangre , Receptores de Lipopolisacáridos/genética , Polimorfismo de Nucleótido Simple/genética , Secuencia de Bases , Estudios de Casos y Controles , Niño , Preescolar , China , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN

RESUMEN

The Yangtze River Delta white goat is a goat breed that can produce high quality brush hair (Type III hair) around the world. This study aimed to compare Type III hair and non-Type III hair goat skin tissues using differentially expressed proteins based on 2-dimensional gel electrophoresis technology. The differentially expressed protein spots were analyzed using the PDquest 8.0 software. Ten protein spots were detected as positive for mass spectrometric analysis based on a threshold of 2-fold change. Through matching based on Ultraflex III TOF/TOF and MASCOT database, four differentially expressed proteins were identified. Fibrinogen beta chain isoform 1 and ATP synthase beta subunit were upregulated in Type III hair, while succinyl-CoA:3-ketoacid-coenzyme A transferase 1-mitochondrial-like and actin-cytoplasmic 1 were upregulated in non-Type III hair. The 4 proteins play important roles in different aspects of hair follicle development. These findings could pave a good foundation for explaining the mechanism of forming Type III hair.

Asunto(s)

Cabras/metabolismo , Proteínas/metabolismo , Proteómica/métodos , Piel/metabolismo , Secuencia de Aminoácidos , Animales , Electroforesis en Gel Bidimensional , Masculino , Espectrometría de Masas , Datos de Secuencia Molecular , Mapeo Peptídico , Ríos

RESUMEN

The enhanced green fluorescent protein (EGFP) pEGFP-N1-P53 eukaryotic expression vector, which contains the human tumor suppressor p53, was constructed and transfected into chicken fibroblast cells and stage-X blastoderm to analyze the transfection efficiency. The complementary DNA of the human p53 gene was cloned by reverse transcription-polymerase chain reaction from human peripheral blood and inserted into the pEGFP-N1 vector by HindIII and BamHI double digestion. The pEGFP-N1-P53 vector was transfected into chicken embryo fibroblasts by Lipofectamine 2000 liposomes, and the transfection efficiency was analyzed by fluorescence microscope after 36 h of transfection. The stage-X blastoderm was also transfected by blastoderm injection using Lipofectamine 2000 liposomes at room temperature after 12-24 h; then hatching occurred until seventh day, and the transfection efficiency was analyzed by fluorescence microscope in the dead embryo. A total of 90 hatching eggs were transfected by the pEGFP-N1-P53 vector, and 20 chicken embryos expressed the reporter gene, which indicated that recombinant pEGFP-N1-P53 could be transfected and expressed in stage-X blastoderm by liposomes. Chicken embryo fibroblasts were transfected and expressed the reporter gene. The pEGFP-N1-P53 vector was constructed successfully and could be transfected and expressed in chicken embryo fibroblasts and stage-X blastoderms efficiently.

Asunto(s)

Vectores Genéticos , Proteínas Fluorescentes Verdes/genética , Proteínas Recombinantes de Fusión/genética , Proteína p53 Supresora de Tumor/genética , Animales , Blastodermo/crecimiento & desarrollo , Blastodermo/metabolismo , Embrión de Pollo , Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Humanos , Proteínas Recombinantes de Fusión/biosíntesis , Proteína p53 Supresora de Tumor/biosíntesis

RESUMEN

The aim of our study was to evaluate the efficacy and safety of two concentrations of botulinum toxin A (BTX-A) for the treatment of hemifacial spasm. We randomly divided 20 patients with hemifacial spasm into high- and low-concentration groups; they were administered 50 and 25 U/mL BTX-A injection, respectively. Further, we compared the curative effects and the occurrence of adverse reactions in the two groups. Our results showed that both the concentrations of BTX-A were effective and no significant difference was observed in the onset time and therapeutic efficacy between the two groups; however, the duration of efficacy was longer in the high-concentration group than in the low-concentration group. Patients in both groups had no allergic reactions and systemic toxic reactions, but those in the high-concentration group had more serious adverse reactions and they lasted for longer durations. The adverse reactions in the two groups were not specifically treated, and they resolved in a relatively short time. In conclusion, local injection of BTX-A was effective in treating hemifacial spasm and each concentration of BTX-A had advantages and disadvantages, which indicated that the concentration of BTX-A can be selected according to the clinical characteristics and willingness of the patients.

Asunto(s)

Toxinas Botulínicas Tipo A/administración & dosificación , Espasmo Hemifacial/tratamiento farmacológico , Adulto , Toxinas Botulínicas Tipo A/efectos adversos , Estudios Cruzados , Método Doble Ciego , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/patología , Femenino , Espasmo Hemifacial/patología , Humanos , Masculino , Persona de Mediana Edad

RESUMEN

Invasion, metastasis, and recurrence are the most common causes of death in patients with hepatocellular carcinoma (HCC) and are therefore critical factors for both therapy and prognosis. Current methods for diagnosis of HCC rely mainly on serological markers such as alpha-fetoprotein and liver enzymes, together with physical assessment and imaging techniques. The availability of more accurate serum markers may facilitate screening and early diagnosis, which will improve prognosis. This retrospective cohort analysis included 50 consecutive patients with cirrhosis and single or multifocal HCC and 40 control subjects with no liver disease or risk factors for viral hepatitis. Expression of epidermal growth factor-like domain 7 (EGFL7), osteopontin (OPN), and prostaglandin E2 (PGE2) were detected using an enzyme-linked immunosorbent assay. The mean serum levels of EGFL7, OPN, and PGE2 in the HCC group were 132.11 pg/mL, 11.77 ng/mL, and 179.37 pg/mL, respectively, which were all significantly higher than the levels in the control group (23.03 pg/mL, 2.31 ng/mL, and 47.36 pg/mL, respectively; P < 0.001). Serum levels of EGFL7, OPN, and PGE2 levels may thus be useful for screening and surveillance of HCC among high-risk populations, and have the potential to improve prognosis of these patients.

Asunto(s)

Carcinoma Hepatocelular/sangre , Carcinoma Hepatocelular/genética , Regulación Neoplásica de la Expresión Génica , Neoplasias Hepáticas/sangre , Neoplasias Hepáticas/genética , Adulto , Anciano , Biomarcadores de Tumor/sangre , Proteínas de Unión al Calcio , Dinoprostona/sangre , Familia de Proteínas EGF , Factores de Crecimiento Endotelial/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Fibrosis/sangre , Perfilación de la Expresión Génica , Marcadores Genéticos/genética , Humanos , Hígado/enzimología , Masculino , Persona de Mediana Edad , Análisis Multivariante , Metástasis de la Neoplasia , Recurrencia Local de Neoplasia , Osteopontina/sangre , Pronóstico , Estudios Retrospectivos , Factores de Riesgo , alfa-Fetoproteínas/metabolismo

RESUMEN

FGD1 encoding a guanine nucleotide exchange factor, specifically activates Rho GTPase cell division cycle 42 (Cdc42). Dysfunction of FGD1 causes Aarskog-Scott syndrome (MIM #305400), an X-linked disorder that may affect bone and intellectual development. However, the relationship between FGD1 and intellectual developmental disorders (IDD) remains unclear. The purpose of this study was to investigate the genetic association between the FGD1 polymorphism and IDD. Working with families from the Qinba mountain area where the occurrence of IDD is higher than the average in China, we analyzed 456 samples from 130 nuclear families, effectively controlling for stratification and environmental factors. Five SNP loci (rs2230265, rs7881608, rs2239809, rs6614244, and rs2284710) were selected that were well distributed within the FGD1 gene. Genotyping was performed through single-strand conformation polymorphism and restriction fragment length polymorphism. The data were analyzed with transmission disequilibrium tests. In the Qinba mountain area, no significant association was observed between IDD and allele or genotype frequencies, or the haplotype of the 5 SNP loci of the FGD1 gene. The results indicate that FGD1 may not be a monogenetic X-linked factor in IDD. Further studies are required to investigate its role in intellectual development based on its specific interactions with Cdc42 or other partner proteins contributing to IDD.

Asunto(s)

Discapacidades del Desarrollo/genética , Factores de Intercambio de Guanina Nucleótido/genética , Polimorfismo de Nucleótido Simple , Adulto , Estudios de Casos y Controles , Niño , Preescolar , China , Humanos

RESUMEN

Type 1 diabetes is a chronic progressive autoimmune disease characterized by mononuclear cell infiltration, with subsequent destruction of insulin-producing ß-cells. Studies have identified strong associations between type 1 diabetes and several chromosome regions, including 12q24. Association between type 1 diabetes and 12q24 arises from SNP rs3184504; rs3184504 is a nonsynonymous SNP in exon 3 of SH2B3 (also known as LNK). Nonobese diabetic (NOD) mice recapitulate many aspects of the pathogenesis of type 1 diabetes in humans and are therefore frequently used in studies addressing the cellular and molecular mechanisms of this disease. It is of interest to know whether there is a similar mutation of SH2B3 in NOD mice. We found that the SH2B3 mutation is absent in NOD mice. To our knowledge, this is the first report of the sequence and the protein levels of SH2B3 in NOD mice.

Asunto(s)

Diabetes Mellitus Tipo 1/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Mutación , Proteínas Adaptadoras Transductoras de Señales , Animales , Secuencia de Bases , Cartilla de ADN , Femenino , Proteínas de la Membrana , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Obesidad , Polimorfismo de Nucleótido Simple

RESUMEN

Calmodulin (CaM) is a multifunctional intracellular calcium ion receptor protein that participates in a range of cellular processes, including calcium metabolism in mussels. To investigate the role of CaM in freshwater mollusk shell calcium metabolism, the full-length CaM cDNA was isolated from the freshwater pearl mussel, Hyriopsis schlegelii (referred to as hsCaM) using SMART RACE technology. The full-length hsCaM was 855 bp in size, containing a 70-bp 5'-untranslated sequence, a 447-bp open reading frame, a 309-bp 3'-untranslated sequence, and a 26-nucleotide long poly(A) tail. The hsCaM mRNA expression in different mussel tissues was examined using real-time PCR. The hsCaM mRNA was found to be ubiquitously expressed, but far more abundant in the gill, foot, and mantle than in the posterior adductor muscle. Real-time PCR was also used to determine hsCaM mRNA expression levels in mantle tissues of H. schlegelii at different ages. No significant differences between one-, two-, and three-year-old mussels were detected, but expression increased in four-year-old mussels and then decreased in five-year-old mussels. CaM appears to be involved in calcium regulation of the mantle in four-year-old mussels, which may secrete more mother of pearl during pearl culture.

Asunto(s)

Bivalvos/genética , Calmodulina/genética , Calmodulina/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Análisis de Secuencia de ADN