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Tunchang pigs, mainly distributed throughout Hainan Province of China, are well-known for their superior meat quality, crude feed tolerance, and adaptability to high temperatures and humidity. Runs of homozygosity (ROH) can provide valuable information about the inbreeding coefficient in individuals and selection signals that may reveal candidate genes associated with key functional traits. Runs of heterozygosity (ROHet) are commonly associated with balance selection, which can help us understand the adaptive evolutionary history of domestic animals. In this study, we investigated ROHs and ROHets in 88 Tunchang pigs. We also compared the estimates of inbreeding coefficients in individuals calculated based on four methods. In summary, we detected a total of 16 ROH islands in our study, and 100 genes were found within ROH regions. These genes were correlated with economically important traits such as reproduction (e.g., SERPIND1, HIRA), meat quality (e.g., PI4KA, TBX1), immunity (e.g., ESS2, RANBP1), adaption to heat stress (TXNRD2 and DGCR8), and crude food tolerance (TRPM6). Moreover, we discovered 18 ROHet islands harbouring genes associated with reproduction (e.g., ARHGEF12, BMPR2), immune system (e.g., BRD4, DNMT3B). These findings may help us design effective breeding and conservation strategies for this unique breed.
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Heterozigoto , Homozigoto , Animais , Suínos/genética , Endogamia , China , Sus scrofa/genética , Feminino , Genoma , MasculinoRESUMO
The Sunong black pig is a new composite breed under development generated from Chinese indigenous pig breeds (i.e., Taihu and Huai) and intensive pig breeds (i.e., Landrace and Berkshire), which is an important genetic material for studying breeding mechanisms. However, there is currently limited knowledge about the genetic structure and germplasm characteristics of Sunong black pigs. To comprehensively understand their genetic composition and ancestry proportions, we performed population structure and local ancestry inference analysis based on whole-genome sequencing information. The results showed that Sunong black pigs could be clustered independently into a group, whose pedigree was intermediate between indigenous and commercial pig breeds, but closer to commercial pigs. Furthermore, local ancestry inference analysis revealed that Sunong black pigs inherited immune and reproductive traits from indigenous pig breeds, including CC and CXC chemokine family, Toll-like receptor family, IFN gene family, ESR1, AREG and EREG gene, while growth and development-related traits were inherited from commercial pig breeds, including IGF1 and GSY2 gene. Overall, Sunong black pigs have formed a relatively stable genome structure with some advantageous traits inherited from their ancestral breeds. This study deepened the understanding of the breeding mechanism of Sunong black pigs and provided a reference for cross-breeding programmes in livestock.
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Polimorfismo de Nucleotídeo Único , Sus scrofa , Suínos/genética , Animais , Sus scrofa/genética , Linhagem , Genoma , Análise de Sequência de DNA/veterinária , Variação GenéticaRESUMO
The biological samples of oral genetic diseases and rare diseases are extremely precious. Collecting and preserving these biological samples are helpful to elucidate the mechanisms and improve the level of diagnose and treatment of oral genetic diseases and rare diseases. The standardized construction of biobanks for oral genetic diseases and rare diseases is important for achieving these goals. At present, there is very little information on the construction of these biobanks, and the standards or suggestions for the classification and coding of biological samples from oral and maxillofacial sources, and this is not conducive to the standardization and information construction of biobanks for special oral diseases. This consensus summarizes the background, necessity, principles, and key points of constructing the biobank for oral genetic diseases and rare diseases. On the base of the group standard "Classification and Coding for Human Biomaterial" (GB/T 39768-2021) issued by the National Technical Committee for Standardization of Biological Samples, we suggest 76 new coding numbers for different of biological samples from oral and maxillofacial sources. We hope the consensus may promote the standardization, and smartization on the biobank construction as well as the overall research level of oral genetic diseases and rare diseases in China.
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Bancos de Espécimes Biológicos , Doenças Raras , Humanos , Doenças Raras/genética , Consenso , ChinaRESUMO
Objective: To explore the mechanisms of prickle planar cell polarity protein 1 (PRICKLE1) involved in the occurrence of skeletal Class â ¢ malocclusion. Methods: After extracting the genomic DNA of all family members of the skeletal Class â ¢ malocclusion pedigree with maxillary hypoplasia collected in the Department of Orthodontics at the Affiliated Stomatological Hospital of Nanjing Medical University in October 2021, whole exome sequencing and Sanger sequencing were performed to screen pathogenic genes/mutation sites and validate the mutations. Jaw tissue was collected during the operation of orthognathic patients who were treated in the Department of Oral and Maxillofacial Surgery at the same hospital from October 2021 to December 2022. Following the extraction of human jaw bone marrow mesenchymal stem cells and transfection with overexpressing lentivirus (lentiviruses overexpressing the gene of interest served as the wild group, lentiviruses overexpressing mutation site served as the mutant group) and knockdown lentivirus (divided into knockdown group 1 and 2, with transfection interference negative lentiviruses as the control group). Various assays including real-time fluorescence quantitative PCR (RT-qPCR), Western blotting, proliferation and Transwell assays, alkaline phosphatase staining and alizarin red staining were performed. Construction of zebrafish animal model, morpholino oligonucleotide (MO) were injected to knock down the expression of prickle1a and prickle1b in zebrafish (co-knocking group), and the control group was injected with standardized MO as a reference. Transcriptome sequencing, enrichment analysis and co-expression analysis were performed on the zebrafish craniofacial tissues of the two groups. Results: Two patients of this family carried this mutation PRICKLE1 c.113C>T. The transfection experiments showed that compared with the wild group (relative expression of PRICKLE1 was 21.97±0.60), the relative expression of mutant group (5.05±0.05) was significantly reduced (P<0.05), and cell proliferation and migration ability significantly enhanced (P<0.05), and osteogenic differentiation ability was significantly reduced (P<0.05). Compared with the control group, the proliferation and migration ability of cells in the two knockdown groups were significantly enhanced (P<0.05), and the osteogenic differentiation ability was significantly reduced (P<0.05). Zebrafish model experiments showed the width of the ethmoid plate was significantly reduced in the co-knocking group (282.50±61.77, t=5.29, P<0.001) compared with the control group (338.80±24.92). Transcriptome data and enrichment analysis showed that the differentially expressed genes were significantly enriched in the mitogen-activated protein kinase (MAPK) signaling pathway after the simultaneous knockdown of prickle1a and prickle1b in zebrafish. Conclusions: PRICKLE1 c.113C>T mutation might suppress the osteoblastic differentiation ability of jaw bone marrow mesenchymal stem cells by downregulating the MAPK signaling pathway, thereby involving the development of skeletal Class â ¢ malocclusion.
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Heat stress is a major problem that constrains pig productivity. Understanding and identifying adaptation to heat stress has been the focus of recent studies, and the identification of genome-wide selection signatures can provide insights into the mechanisms of environmental adaptation. Here, we generated whole-genome re-sequencing data from six Chinese indigenous pig populations to identify genomic regions with selection signatures related to heat tolerance using multiple methods: three methods for intra-population analyses (Integrated Haplotype Score, Runs of Homozygosity and Nucleotide diversity Analysis) and three methods for inter-population analyses (Fixation index (FST), Cross-population Composite Likelihood Ratio and Cross-population Extended Haplotype Homozygosity). In total, 1 966 796 single nucleotide polymorphisms were identified in this study. Genetic structure analyses and FST indicated differentiation among these breeds. Based on information on the location environment, the six breeds were divided into heat and cold groups. By combining two or more approaches for selection signatures, outlier signals in overlapping regions were identified as candidate selection regions. A total of 163 candidate genes were identified, of which, 29 were associated with heat stress injury and anti-inflammatory effects. These candidate genes were further associated with 78 Gene Ontology functional terms and 30 Kyoto Encyclopedia of Genes and Genomes pathways in enrichment analysis (P < 0.05). Some of these have clear relevance to heat resistance, such as the AMPK signalling pathway and the mTOR signalling pathway. The results improve our understanding of the selection mechanisms responsible for heat resistance in pigs and provide new insights of introgression in heat adaptation.
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Termotolerância , Suínos/genética , Animais , Seleção Genética , Genoma , Homozigoto , Haplótipos , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Diabetic foot wound repair is a challenging issue in clinical practice. Due to the influence of multiple factors including the damage and regeneration failure of local tissue, the impaired pathways of wound repairing through blood vessels and nerve nutrition, and disorders of a variety of cellular factors, traditional treatment methods are often difficult to achieve good therapeutic effects. Stem cells are a type of cells with potentials of multidirectional differentiation, which also possess functions such as regulating immunity and paracrine to facilitate the comprehensive wound repair, so they have promising application prospect at present for the treatment of diabetic foot wounds. Because the relevant parameters of stem cell treatment are in the exploratory phase, there were no standardized data. This paper reviews the application of stem cells in the research of diabetic foot wound treatment over the past 6 years, analyzing and summarizing the contents in focused aspects including the types and sources of stem cells, effects of donor age and gender on stem cells, mode of administration, transplantation survival rate and safety, which may provide a reference for further application of stem cells in the clinical treatment of diabetic foot wound.
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Diabetes Mellitus , Pé Diabético , Diferenciação Celular , Pé Diabético/terapia , Humanos , Transplante de Células-Tronco , Cicatrização/fisiologiaRESUMO
Objective: To investigate the influence factors of poor efficacy after flap repair operation in patients with pressure ulcers. Methods: The retrospective case series study was conducted. From January 2011 to June 2021, 125 patients with stage â ¢ and â £ pressure ulcers treated in Hainan General Hospital met the inclusion criteria. There were 82 males and 43 females, aged 15-90 (57±20) years. According to the postoperative effects, the patients were divided into poor efficacy group (47 cases) and good efficacy group (78 cases). The clinical data of patients in the two groups were collected, including the age, gender, location, stage, size, and bone exposure of pressure ulcers, preoperative microorganism culture results of wound exudate sample, whether combined with osteomyelitis, diabetes, lower limb paroxysmal myospasm, and gatism or not, the number of surgical debridement combined with negative-pressure wound therapy, type of surgical flap, postoperative position, and preoperative albumin, leukocyte, C-reactive protein (CRP), and hemoglobin. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, and chi-square test. The binary multivariate logistic regression analysis was conducted to screen the independent risk factors influencing the poor efficacy after flap repair operation in 125 patients with stage â ¢ and â £ pressure ulcers. Results: The ratio of patients with lower limb paroxysmal myospasm in poor efficacy group was 22/47, which was significantly higher than 3/78 in good efficacy group (χ2=33.83, P<0.01). The preoperative hemoglobin level of patients in poor efficacy group was (102±17) g/L, which was significantly lower than (113±20) g/L in good efficacy group (t=-3.24, P<0.01). The preoperative CRP level of patients was 39.1 (14.1, 91.6) mg/L in poor efficacy group, which was significantly higher than 15.3 (6.6, 42.0) mg/L in good efficacy group (Z=-3.04, P<0.01). There were no statistically significant differences in other indexes between patients in the two groups (P>0.05). Multivariate logistic regression analysis showed that age, lower limb paroxysmal myospasm, and preoperative hemoglobin level were the independent risk factors for poor efficacy after flap repair operation in patients with pressure ulcers (with odds ratios of 1.03, 40.69, and 0.97, 95% confidence intervals of 1.00-1.06, 9.18-180.39, and 0.95-1.00, respectively, P<0.05 or P<0.01). Conclusions: Poor efficacy after flap repair operation in patients with pressure ulcers is affected by many factors, among which the age, lower limb paroxysmal myospasm, and preoperative hemoglobin level are the independent risk factors.
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Diabetes Mellitus , Úlcera por Pressão , Masculino , Feminino , Humanos , Estudos Retrospectivos , Úlcera por Pressão/cirurgia , Retalhos Cirúrgicos , Transplante de Pele , Resultado do TratamentoRESUMO
OBJECTIVE: To explore the relationships between hepatitis B virus (HBV) DNA, HBV RNA and hepatitis B surface antigen (HBsAg) and to evaluate their predictive value for mother-to-child transmission of HBV. DESIGN: An observational cohort study. SETTING: First Hospital of Jilin University. POPULATION: HBsAg-positive and hepatitis B e antigen (HBeAg) -positive pregnant women were recruited. METHODS: Blood samples were collected from mothers before delivery, and HBV infection of infants was evaluated at 7 months of age. RESULTS: Overall, 268 mothers and 271 infants were enrolled. HBV DNA and HBsAg levels were correlated (rs = 0.699; P < 0.001), and HBV DNA (rs = 0.500; P < 0.001) and HBsAg (rs = 0.372; P < 0.001) were both correlated with HBV RNA. The areas under the curve for HBV DNA, HBsAg and HBV RNA for prediction of infection were 0.69 (95% CI 0.57-0.82), 0.63 (95% CI 0.51-0.76) and 0.65 (95% CI 0.52-0.78), respectively. Higher HBV DNA (odds ratio [OR] 4.77, 95% CI 1.44-15.86), higher HBsAg (OR 4.13, 95% CI 1.12-15.25) and higher HBV RNA (OR 3.19, 95% CI 1.09-9.32) were risk factors for HBV infection. Analysis of the HBV DNA-RNA-HBsAg Score revealed that it was an independent predictive factor for mother-to-child transmission (the OR of Score 3 was 8.81, 95% CI 2.79-27.82). CONCLUSION: HBV DNA, HBV RNA and HBsAg were correlated in HBeAg-positive pregnant women. HBsAg could be considered as a substitute marker of HBV DNA for HBeAg-positive pregnant women in low-income regions. We should pay special attention to pregnant women with high levels of all three markers. TWEETABLE ABSTRACT: HBsAg could be considered as a substitute marker of HBV DNA for HBeAg-positive pregnant women in low-income regions. Special attention should be given to pregnant women with high levels of all three markers (HBV DNA, HBV RNA and HBsAg).
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Vírus da Hepatite B/isolamento & purificação , Hepatite B/transmissão , Transmissão Vertical de Doenças Infecciosas , Complicações Infecciosas na Gravidez/virologia , Adulto , Estudos de Coortes , DNA Viral , Feminino , Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Humanos , Lactente , Recém-Nascido , Masculino , Valor Preditivo dos Testes , Gravidez , Estudos Prospectivos , RNA Viral , Estudos Retrospectivos , Carga ViralRESUMO
Avibacterium paragallinarum has been subtyped into three serogroups (A, B, and C) and nine serovars (A-1, A-2, A-3, A-4, B-1, C-1, C-2, C-3, and C-4) according to the Page and Kume schemes. Both schemes use the hemagglutination inhibition test for serotyping. However, the relationship between the hemagglutinin gene (HMTp210) sequences and serotypes of A. paragallinarum is still unclear. This problem is partly due to the lack of information on the complete HMTp210 sequence from the formal reference strain of Page serogroup B (strain 0222 or Spross). In this study, we determined the complete HMTp210 sequence of strain Spross. The sequence of Spross and those of other HMTp210 sequences retrieved from GenBank were used to conduct phylogenetic analyses to investigate the relationship between the serotypes and HMTp210 sequences of A. paragallinarum. Four phylogenetic clusters, designated clusters A-1, A-2, B, and C, were identified. Clustering based on complete HMTp210 sequences correlates with serotyping based on hemagglutination inhibition tests. Serovar A-2 was found to contain a chimeric HMTp210 gene that might have resulted from recombination between serovar A-1 and serovar C-1. In addition, phylogenetic analysis based on partial sequences (approximately nucleotides 1-1200) of HMTp210 was sufficient to discriminate between serogroups A, B, and C. These findings could be valuable for developing a molecular method for serotyping of A. paragallinarum.
Relación entre los serotipos y las secuencias génicas de hemaglutinina de Avibacterium paragallinarum. Avibacterium paragallinarum se ha subtipificado en tres serogrupos (A, B y C) y nueve serovares (A-1, A-2, A-3, A-4, B-1, C-1, C-2, C- 3 y C-4) de acuerdo con los esquemas Page y Kume. Ambos esquemas utilizan la prueba de inhibición de la hemaglutinación para la serotipificación. Sin embargo, la relación entre las secuencias del gene de la hemaglutinina (HMTp210) y los serotipos de A. paragallinarum aún no está clara. Este problema se debe en parte a la falta de información sobre la secuencia completa del gene HMTp210 de la cepa de referencia formal del serogrupo B de Page (cepa 0222 o Spross). En este estudio, se determinó la secuencia completa de HMTp210 de la cepa Spross. La secuencia de Spross y las de otras secuencias del gene HMTp210 obtenidas de GenBank se utilizaron para realizar análisis filogenéticos para investigar la relación entre los serotipos y las secuencias de HMTp210 de A. paragallinarum. Se identificaron cuatro agrupaciones filogenéticas, denominadas grupos A-1, A-2, B y C. La agrupación basada en las secuencias completas del gene HMTp210 se correlaciona con la serotipificación basada en pruebas de inhibición de la hemaglutinación. Se encontró que el serovar A-2 contenía un gene HMTp210 quimérico que podría haber resultado de la recombinación entre el serovar A-1 y el serovar C-1. Además, el análisis filogenético basado en secuencias parciales (aproximadamente nucleótidos 1-1200) del gene HMTp210 fue suficiente para discriminar entre los serogrupos A, B y C. Estos hallazgos podrían ser valiosos para desarrollar un método molecular para la serotipificación de A. paragallinarum.
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Infecções por Haemophilus , Haemophilus paragallinarum , Doenças das Aves Domésticas , Animais , Galinhas , Infecções por Haemophilus/veterinária , Haemophilus paragallinarum/genética , Hemaglutininas/genética , Filogenia , SorogrupoRESUMO
On 17 August 2017, the Laser Interferometer Gravitational-Wave Observatory (LIGO) and the Virgo interferometer detected gravitational waves (GWs) emanating from a binary neutron star merger, GW170817. Nearly simultaneously, the Fermi and INTEGRAL (INTErnational Gamma-Ray Astrophysics Laboratory) telescopes detected a gamma-ray transient, GRB 170817A. At 10.9 hours after the GW trigger, we discovered a transient and fading optical source, Swope Supernova Survey 2017a (SSS17a), coincident with GW170817. SSS17a is located in NGC 4993, an S0 galaxy at a distance of 40 megaparsecs. The precise location of GW170817 provides an opportunity to probe the nature of these cataclysmic events by combining electromagnetic and GW observations.
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Eleven hours after the detection of gravitational wave source GW170817 by the Laser Interferometer Gravitational-Wave Observatory and Virgo Interferometers, an associated optical transient, SSS17a, was identified in the galaxy NGC 4993. Although the gravitational wave data indicate that GW170817 is consistent with the merger of two compact objects, the electromagnetic observations provide independent constraints on the nature of that system. We synthesize the optical to near-infrared photometry and spectroscopy of SSS17a collected by the One-Meter Two-Hemisphere collaboration, finding that SSS17a is unlike other known transients. The source is best described by theoretical models of a kilonova consisting of radioactive elements produced by rapid neutron capture (the r-process). We conclude that SSS17a was the result of a binary neutron star merger, reinforcing the gravitational wave result.
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On 17 August 2017, Swope Supernova Survey 2017a (SSS17a) was discovered as the optical counterpart of the binary neutron star gravitational wave event GW170817. We report time-series spectroscopy of SSS17a from 11.75 hours until 8.5 days after the merger. Over the first hour of observations, the ejecta rapidly expanded and cooled. Applying blackbody fits to the spectra, we measured the photosphere cooling from [Formula: see text] to [Formula: see text] kelvin, and determined a photospheric velocity of roughly 30% of the speed of light. The spectra of SSS17a began displaying broad features after 1.46 days and evolved qualitatively over each subsequent day, with distinct blue (early-time) and red (late-time) components. The late-time component is consistent with theoretical models of r-process-enriched neutron star ejecta, whereas the blue component requires high-velocity, lanthanide-free material.
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On 17 August 2017, gravitational waves (GWs) were detected from a binary neutron star merger, GW170817, along with a coincident short gamma-ray burst, GRB 170817A. An optical transient source, Swope Supernova Survey 17a (SSS17a), was subsequently identified as the counterpart of this event. We present ultraviolet, optical, and infrared light curves of SSS17a extending from 10.9 hours to 18 days postmerger. We constrain the radioactively powered transient resulting from the ejection of neutron-rich material. The fast rise of the light curves, subsequent decay, and rapid color evolution are consistent with multiple ejecta components of differing lanthanide abundance. The late-time light curve indicates that SSS17a produced at least ~0.05 solar masses of heavy elements, demonstrating that neutron star mergers play a role in rapid neutron capture (r-process) nucleosynthesis in the universe.
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Hypoxia plays a critical role during the evolution of malignant cells and tumour microenvironment (TME).Tumour-derived exosomes contain informative microRNAs involved in the interaction of cancer and stromal cells, thus contributing to tissue remodelling of tumour microenvironment. This study aims to clarify how hypoxia affects tumour angiogenesis through exosomes shed from lung cancer cells. Lung cancer cells produce more exosomes under hypoxic conditions than do parental cells under normoxic conditions. miR-23a was significantly upregulated in exosomes from lung cancer under hypoxic conditions. Exosomal miR-23a directly suppressed its target prolyl hydroxylase 1 and 2 (PHD1 and 2), leading to the accumulation of hypoxia-inducible factor-1 α (HIF-1 α) in endothelial cells. Consequently, hypoxic lung cancer cells enhanced angiogenesis by exosomes derived from hypoxic cancer under both normoxic and hypoxic conditions. In addition, exosomal miR-23a also inhibits tight junction protein ZO-1, thereby increasing vascular permeability and cancer transendothelial migration. Inhibition of miR-23a by inhibitor administration decreased angiogenesis and tumour growth in a mouse model. Furthermore, elevated levels of circulating miR-23a are found in the sera of lung cancer patients, and miR-23a levels are positively correlated with proangiogenic activities. Taken together, our study reveals the clinical relevance and prognostic value of cancer-derived exosomal miR-23a under hypoxic conditions, and investigates a unique intercellular communication, mediated by cancer-derived exosomes, which modulates tumour vasculature.
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Permeabilidade Capilar/fisiologia , Exossomos/metabolismo , Neoplasias Pulmonares/metabolismo , MicroRNAs/metabolismo , Neovascularização Patológica/metabolismo , Prolil Hidroxilases/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo , Animais , Hipóxia Celular/fisiologia , Linhagem Celular , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Hipóxia/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas de Junções Íntimas/metabolismoRESUMO
OBJECTIVE: Tooth agenesis is a common craniofacial anomaly in human beings. Mounting evidence has demonstrated that the bone morphogenetic protein 4 gene (BMP4) plays an important role in tooth development. This case-control study was designed to evaluate the association of the polymorphism rs17563 in BMP4 gene with susceptibility of isolated human tooth agenesis in a Chinese Han population. PATIENTS AND METHODS: 335 tooth agenesis cases and 444 healthy controls were included in this study. RESULTS: Although no significant association was observed either in the overall or stratified analysis between the types and the severity of missing teeth. However, significant difference was observed between the anterior and posterior tooth agenesis (APTA) cases and the controls (p = 0.018 for allele distribution and OR = 0.39, 95% CI = 0.15-0.99). Furthermore, the heterozygote (TC) and dominant model (CC+TC) were associated with decreased risk of APTA compared with the control (phet = 0.018, ORhet = 0.39, 95% CIhet = 0.15-0.99 and pdom = 0.042, ORdom = 0.34, 95% CIdom = 0.13-0.87, respectively). CONCLUSIONS: These results indicated that rs17563 in BMP4 gene was potentially associated with APTA in Chinese Han population and further independent studies are required to verify these findings.
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Alelos , Anodontia/genética , Povo Asiático/genética , Proteína Morfogenética Óssea 4/genética , Polimorfismo de Nucleotídeo Único/genética , Vigilância da População , Adolescente , Adulto , Anodontia/sangue , Anodontia/etnologia , Povo Asiático/etnologia , Proteína Morfogenética Óssea 4/sangue , Estudos de Casos e Controles , Criança , Feminino , Estudos de Associação Genética/métodos , Humanos , Masculino , Vigilância da População/métodos , Adulto JovemRESUMO
As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. Yeast CUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study, CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.
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Animais , Humanos , Mamíferos/fisiologia , Feromônios/fisiologia , Comportamento Animal/fisiologia , Comportamento/fisiologia , Odorantes , Bulbo Olfatório/fisiologia , Mucosa Olfatória/fisiologia , Condutos Olfatórios/anatomia & histologia , Condutos Olfatórios/fisiologia , Neurônios Receptores Olfatórios/fisiologia , Feromônios Humano/fisiologia , Olfato/fisiologiaRESUMO
As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. Yeast CUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study, CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.
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Cobre/metabolismo , Metalotioneína/fisiologia , Estresse Oxidativo/fisiologia , Proteínas de Transporte/análise , Proteínas de Transporte/metabolismo , Ciclo Celular/fisiologia , Sobrevivência Celular/fisiologia , Cobre/análise , Formazans , Células HeLa , Humanos , Metalotioneína/análise , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Sais de Tetrazólio , Fatores de TempoRESUMO
OBJECTIVE: To evaluate the effect of midpalatal mini-implant anchorage system in fixed appliance treatment. METHODS: In this study, 14 adolescents who had skeletal class I or II malocclusions were involved. Maximal anchorage was required during orthodontic treatment. Maxillary first premolars of the selected individuals were extracted and the individuals were treated by fixed appliance. One mini-implant was inserted in the midpalatal suture region and a transpalatal arch (TPA) made of 0.019 inch×0.022 inch(1 inch=2.54 cm) stainless steel was adhered to the mini-implant and upper first molars. Cephalometric radiographs taken after mini-implants inserted (T0) and before mini-implants removing (T1) were traced and measured. SN-7 plane and PP plane were used as reference planes. Student's t-test was used. RESULTS: The successful rate of midpalatal mini-implant was 73.9%. All the items measured were found with no significant difference between the two groups. CONCLUSION: This mini-implant system as orthodontic anchorage in midpalatal region can be an alternative method of maximal anchorage during orthodontic treatment.
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Procedimentos de Ancoragem Ortodôntica , Técnicas de Movimentação Dentária , Adolescente , Dente Pré-Molar , Cefalometria , Humanos , Maxila , Dente Molar , Resultado do TratamentoRESUMO
OBJECTIVE: To examine whether periostin-like factor (PLF) stimulation of the expanded palatal suture would accelerate mineral formation of bone. MATERIALS AND METHODS: Expanded palates of 6-week-old male C57BL/6 mice were maintained in an organ culture system, and tissue was either unstimulated or stimulated with PLF or BMP-7 for 8 days. Bone mineral formation was assessed by Raman microspectroscopy analysis and histological examination. RESULTS: Raman microspectroscopy analysis demonstrated that unstimulated palates maintained their bone mineral concentration within the palatal suture over 8 days (%increase = 11.29 ± 2.34). In comparison, palates exposed to either PLF (%increase = 29.33 ± 1.61, p = 0.012) or BMP-7 (% increase = 25.49 ± 1.09, p = 0.016) formed significantly more bone at the osteogenic fronts of the palatal suture compared with unstimulated samples at day 8. Alkaline phosphatase activity along the bone edges was markedly greater in the PLF and BMP-7 groups compared with that in unstimulated groups at day 0 and day 8. The levels of osteocalcin proteins in the palatal suture tissues of the PLF and BMP-7 groups were significantly higher than those in the unstimulated group. CONCLUSION: PLF can increase bone mineral formation within the expanded palatal suture.
Assuntos
Moléculas de Adesão Celular/farmacologia , Suturas Cranianas/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Técnica de Expansão Palatina , Palato/efeitos dos fármacos , Fosfatase Alcalina/análise , Animais , Apatitas/análise , Biomarcadores/análise , Western Blotting , Matriz Óssea/efeitos dos fármacos , Proteína Morfogenética Óssea 7/farmacologia , Calcificação Fisiológica/efeitos dos fármacos , Colágeno/análise , Suturas Cranianas/patologia , Durapatita/análise , Módulo de Elasticidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microespectrofotometria , Técnicas de Cultura de Órgãos , Osteocalcina/análise , Palato/patologia , Fosfatos/análise , Distribuição Aleatória , Análise Espectral Raman , Fatores de TempoRESUMO
The cytosolic activity of glycerol-3-phosphate dehydrogenase 1 (GPD1, EC 1.1.1.8) plays an important role in the synthesis of triacylglycerol and in the transport of reducing equivalents from the cytosol to mitochondria. Here we report the full-length genomic sequence of porcine GPD1 gene including promoter region. Porcine GPD1 gene contains eight exons and seven introns. Using the ImpRH, the GPD1 gene was mapped on chromosome 5. Sub-cellular localization of the pig GPD1 was localized in cytoplasm by GFP reporter gene. The full-length CDS of porcine GPD1 gene comprises 1050 nucleotides and it encodes 349 amino acids. Using the CDS sequences of 17 species, we built the phylogeny tree of GPD1 gene. We investigated the expression level of the gene in 13 different tissues and time course from birth to postnatal day 28 in longissinus doris muscle (LD) and in cerebrum. The result shows that porcine GPD1 gene is expressed in almost all tissues we tested but its levels of expression varies widely over 2 orders of magnitude. LD and the cerebrum have similar expression pattern that is at a low level at birth and increasing with aging to the highest level at postnatal day 8 in LD and postnatal day 14 in cerebrum. But weaning decreased the expression level of the GPD1 gene. This may partially explains the effects of weaning on energy metabolism.