RESUMO
Kinship testing of a father and his son from Guangxi, China, the location of the Zhuang minority people, was performed using the PowerPlex® 18D System with a short tandem repeat typing kit. The results indicated that both the father and his son had an off-ladder allele at the Penta D locus, with a genetic size larger than that of the maximal standard allelic ladder. To further identify this locus, monogenic amplification, gene cloning, and genetic sequencing were performed. Sequencing analysis demonstrated that the fragment size of the Penta D-OL locus was 469 bp and the core sequence was [AAAGA]21, also called Penta D-21. The rare Penta D-21 allele was found to be distributed among the Zhuang population from the Guangxi Zhuang Autonomous Region of China; therefore, this study improved the range of DNA data available for this locus and enhanced our ability for individual identification of gene loci.
Assuntos
Variação Genética , Repetições de Microssatélites/genética , Alelos , China , Clonagem Molecular , Eletroforese Capilar , Loci Gênicos , Humanos , Oligossacarídeos , Análise de Sequência de DNARESUMO
This study aimed at comparing the growth and mineralization of the femur, tibia, and metatarsus of male and female broiler chicks. On the day of hatch, 100 male and 100 female Ross 308 broiler chicks were transferred stainless cages with 10 birds per cage. On d 7, 14, 21, 28, 35, and 42, five males and five females were sacrificed and their femur, tibia, and metatarsus were collected. Results showed that the tibia was the heaviest and the longest and contained the highest content of ash and calcium (Ca) among the three leg bones. The femur had the greatest diameter. The weight, length, diameter, and ash weight of the femur, tibia, and metatarsus linearly increased with age. The ash, Ca, and phosphorus (P) content in the femur and the tibia quadratically increased with age; by contrast, these parameters in the metatarsus linearly increased with age. The bones grew faster in 1 to 21 d of age. The weight, diameter, and ash weight of the three bones of males were higher than those of females. The Ca to P ratio of the three bones (femur, tibia, and metatarsus) was approximately 2.0:1. These data indicate that there are differences in bone growth and mineralization among the femur, tibia, and metatarsus of male or female broiler chicks.(AU)
Assuntos
Animais , Galinhas/anatomia & histologia , Galinhas/crescimento & desenvolvimento , Fêmur/anatomia & histologia , Tíbia/anatomia & histologia , Hallux Varus/veterinária , Calcificação FisiológicaRESUMO
This study aimed at comparing the growth and mineralization of the femur, tibia, and metatarsus of male and female broiler chicks. On the day of hatch, 100 male and 100 female Ross 308 broiler chicks were transferred stainless cages with 10 birds per cage. On d 7, 14, 21, 28, 35, and 42, five males and five females were sacrificed and their femur, tibia, and metatarsus were collected. Results showed that the tibia was the heaviest and the longest and contained the highest content of ash and calcium (Ca) among the three leg bones. The femur had the greatest diameter. The weight, length, diameter, and ash weight of the femur, tibia, and metatarsus linearly increased with age. The ash, Ca, and phosphorus (P) content in the femur and the tibia quadratically increased with age; by contrast, these parameters in the metatarsus linearly increased with age. The bones grew faster in 1 to 21 d of age. The weight, diameter, and ash weight of the three bones of males were higher than those of females. The Ca to P ratio of the three bones (femur, tibia, and metatarsus) was approximately 2.0:1. These data indicate that there are differences in bone growth and mineralization among the femur, tibia, and metatarsus of male or female broiler chicks.
Assuntos
Animais , Fêmur/anatomia & histologia , Galinhas/anatomia & histologia , Galinhas/crescimento & desenvolvimento , Hallux Varus/veterinária , Tíbia/anatomia & histologia , Calcificação FisiológicaRESUMO
This study evaluated the effects of dietary non-phytate phosphorus (NPP) and 1a-hydroxycholecalciferol (1a-OH-D3) on the growth performance, bone mineralization, and carcass traits of 1- to 21-day-old broiler chickens. On the day of hatch, 600 male Ross 308 chicks were weighed and randomly assigned to 12 treatments, with five cages of 10 birds each. A 6 × 2 factorial arrangement was applied, consisting of 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, or 0.45% NPP and 0 or 5 g/kg of 1a-OH-D3. The basal diet contained 0.52% calcium (Ca) and was not supplemented with vitamin D3. Dietary NPP levels significantly affected growth performance and tibia mineralization (except width) of broilers; by contrast, meat yield and organ relative weight were not influenced by NPP. The inclusion of 1a-OH-D3 improved growth performance, tibia mineralization, and carcass and breast yield, whereas it decreased the relative weights of the liver, heart, and kidney. A significant interaction between NPP and 1a-OH-D3 was observed for body weight gain (BWG), feed efficiency (FE), mortality, serum Ca and P levels, tibia breaking-strength, ash weight, and Ca content, as well as breast yield and heart relative weight. These results suggest that broilers fed with 5 g of 1a-OH-D3 per kg of diet obtain optimal growth performance and tibia mineralization when dietary NPP level was 0.30% and the analyzed Ca to NPP ratio was 1.97.
Assuntos
Animais , Aves Domésticas/crescimento & desenvolvimento , Carne/análise , Carne/efeitos adversos , Vitamina D/análise , Vitamina D/análogos & derivados , Vitamina D/efeitos adversosRESUMO
This study evaluated the effects of dietary non-phytate phosphorus (NPP) and 1a-hydroxycholecalciferol (1a-OH-D3) on the growth performance, bone mineralization, and carcass traits of 1- to 21-day-old broiler chickens. On the day of hatch, 600 male Ross 308 chicks were weighed and randomly assigned to 12 treatments, with five cages of 10 birds each. A 6 × 2 factorial arrangement was applied, consisting of 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, or 0.45% NPP and 0 or 5 g/kg of 1a-OH-D3. The basal diet contained 0.52% calcium (Ca) and was not supplemented with vitamin D3. Dietary NPP levels significantly affected growth performance and tibia mineralization (except width) of broilers; by contrast, meat yield and organ relative weight were not influenced by NPP. The inclusion of 1a-OH-D3 improved growth performance, tibia mineralization, and carcass and breast yield, whereas it decreased the relative weights of the liver, heart, and kidney. A significant interaction between NPP and 1a-OH-D3 was observed for body weight gain (BWG), feed efficiency (FE), mortality, serum Ca and P levels, tibia breaking-strength, ash weight, and Ca content, as well as breast yield and heart relative weight. These results suggest that broilers fed with 5 g of 1a-OH-D3 per kg of diet obtain optimal growth performance and tibia mineralization when dietary NPP level was 0.30% and the analyzed Ca to NPP ratio was 1.97.(AU)
Assuntos
Animais , Aves Domésticas/crescimento & desenvolvimento , Carne/efeitos adversos , Carne/análise , Vitamina D/análogos & derivados , Vitamina D/efeitos adversos , Vitamina D/análiseRESUMO
The cause of postoperative failure after the treatment of femoral intertrochanteric fracture with proximal femoral nail antirotation (PFNA) was analyzed, and the reoperative methods were examined. Nine failures of 308 femoral intertrochanteric fracture patients with PFNA were treated with femoral prosthesis total hip replacement (THR) and reoperative internal fixation. All nine patients were analyzed to determine the cause of failure. The causes of failed internal fixation in the intertrochanteric-fractured patients included perforation of the helical blade into the hip joint in three cases, cutting-out of the helical blade exit outside in two cases, and hip varus as a result of cutting-out the helical blade in two cases. Seven patients with failed internal fixation were treated with THR. Two patients who had femoral shaft fractures at the end of the nail were treated with longer PFNA. Faulty operative procedures, unsatisfactory reductions, serious osteoporosis, and incorrect positioning of the helical blade were the most important factors responsible for the failed internal fixation. Satisfactory results were achieved with THR and refixation relative to the causes of the failed internal fixation.
Assuntos
Fixação Interna de Fraturas , Fraturas do Quadril/cirurgia , Idoso , Idoso de 80 Anos ou mais , Feminino , Fixação Interna de Fraturas/métodos , Fraturas do Quadril/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Ortopédicos , Cuidados Pós-Operatórios , Cuidados Pré-Operatórios , Retratamento , Falha de Tratamento , Resultado do TratamentoRESUMO
We investigated the polymorphisms of PRLR and FOLR1 genes in Xinong Saanen, Guanzhong, and Boer goat breeds by DNA sequencing and PCR-RFLP. Two novel SNPs were identified: KC109741: g.62130C>T in the 3ê-UTR of goat gene PRLR, and KC136296: g.7884A>C in exon 3 of goat gene FOLR1. In the three goat breeds, the polymorphism information content was 0.20-0.27 at the g.62130C>T locus. At the g.7884A>C locus, it was 0.36 in Boer goats. The three goat breeds were in Hardy-Weinberg disequilibrium at the g.62130C>T locus. The g.62130C>T SNP was found to be significantly associated with milk production traits in Xinong Saanen and Guanzhong breeds. These results are consistent with the regulatory function of PRLR in mammary gland development, milk secretion, and expression of milk protein genes; they extend the spectrum of genetic variation of the goat PRLR gene, which could be useful for breeding programs.
Assuntos
Receptor 1 de Folato/genética , Estudos de Associação Genética , Leite , Receptores da Prolactina/genética , Regiões 3' não Traduzidas , Animais , Cruzamento , Genótipo , Cabras/genética , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNARESUMO
We successfully cloned the Wistar-Imamichi (WI) rat growth hormone gene (GenBank accession: GQ890681), which contained 5 exons and 4 introns. Using the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique, a novel missense substitution single nucleotide polymorphism was identified and tested for Hardy-Weinberg equilibrium with the χ(2) test. This gene fragment was investigated in 50 adult rats and 50 neonatal rats by PCR-SSCP. Results showed that only intron 4 had a polymorphic locus at the 97th nucleotide position from A (in the AA genotype) to C (in the BB genotype). Further statistical analysis showed that this locus was in Hardy-Weinberg equilibrium. These results suggested that this specific pathogen-free WI rat population, which was bred in the barrier system of the Research Center for Laboratory Animal Science of Inner Mongolia University, has high hereditary stability.
Assuntos
Hormônio do Crescimento/genética , Polimorfismo Conformacional de Fita Simples , Animais , Sequência de Bases , Clonagem Molecular , Frequência do Gene , Genótipo , Íntrons , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Ratos , Ratos Wistar , Análise de Sequência de DNARESUMO
Kisspeptins, the product of the KISS1 gene, play an essential role in the regulation of reproductive functions, acting primarily at the hypothalamic level of the gonadotropic axis. We detected polymorphisms of the goat KISS1 gene in 723 individuals from three goat breeds (Xinong Saanen, Guanzhong, and Boer) by DNA pooling, PCR-RFLP, and DNA sequencing methods. We cloned the promoter sequence of this gene and found it to share high similarity with that of the bovine KISS1 promoter. Six TATA boxes were found in the goat KISS1 promoter region. Two novel SNPs (g.2124T>A and g.2270C>T) were identified in the intron 1 of the KISS1 gene of all three goat breeds. The three goat breeds were in Hardy-Weinberg disequilibrium at g.2124T>A and g.2270C>T loci. The g.2124T>A and g.2270C>T loci were closely linked in the three goat breeds (r2 > 0.33). The g.2124T>A and g.2270C>T SNPs were significantly associated with litter size, and the C1 female goats had a larger litter size than did those with the other genotypes. These results extend the spectrum of genetic variation of the goat KISS1 gene, which contributes to our knowledge of goat genetic resources for breeding programs.
Assuntos
Cabras/genética , Kisspeptinas/genética , Tamanho da Ninhada de Vivíparos/genética , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Animais , Clonagem Molecular , Feminino , Frequência do Gene , Estudos de Associação Genética , Loci Gênicos , Genótipo , Desequilíbrio de Ligação , Mutação Puntual , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNARESUMO
Complementary DNA (cDNA) is valuable for investigating protein structure and function in the study of life science, but it is difficult to obtain by traditional reverse transcription. We employed a novel strategy to clone human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA, which was directly isolated from the mucous membrane of mouth. The hLIF sequence, which is 609 bp long and is composed of three exons, can be acquired within a few hours by amplifying each exon and splicing all of them using overlap-PCR. This new approach developed is simple, time- and cost-effective, without RNA preparation or cDNA synthesis, and is not limited to the specific tissues for a particular gene and the expression level of the gene.
Assuntos
Clonagem Molecular/métodos , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Genoma Humano/genética , Fator Inibidor de Leucemia/genética , Mucosa Bucal/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Éxons/genética , Humanos , Fator Inibidor de Leucemia/química , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Splicing de RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Signal transducer and activator of transcription protein 3 (STAT3) has been implicated in cancer development and is recognized as a type of oncogene. However, association studies of single nucleotide polymorphisms (SNPs) in the STAT3 gene with cancer risk are rare and not available for lung cancer. We examined whether STAT3 polymorphisms are associated with the risk of non-small cell lung cancer (NSCLC). Eight SNPs in the STAT3 gene were genotyped by TaqMan assays in 326 NSCLC cases and 432 controls in a Chinese population. Significant decreased risk of NSCLC was observed for carriers of minor alleles rs4796793 (odds ratio (OR) = 0.68, 95% confidence interval (CI) = 0.51-0.92), rs7211777 (OR = 0.67, 95%CI = 0.50-0.90), rs12949918 (OR = 0.73, 95%CI = 0.54-0.97), rs744166 (OR = 0.69, 95%CI = 0.51-0.92), rs9912773 (OR = 0.75, 95%CI = 0.55-0.98), and rs3869550 (OR = 0.70, 95%CI = 0.53-0.94). The GGCGGC haplotype, comprised of minor alleles of the six NSCLC-associated SNPs, had a 0.78-fold (95%CI = 0.62-0.97) significantly decreased risk of NSCLC, as compared to the most common haplotype of CATACT. Stratification analyses by clinical stage showed that the trend for the association between STAT3 polymorphisms and NSCLC risk was present both for stage I/II and stage III/IV, and appeared moderately stronger for stage III/IV. We conclude that polymorphisms in the STAT3 gene may have a protective role in the development of NSCLC, particular of stage III/IV NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Predisposição Genética para Doença , Neoplasias Pulmonares/genética , Fator de Transcrição STAT3/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , China , Feminino , Estudos de Associação Genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Risco , Fatores de Risco , Transdução de Sinais/genética , FumarRESUMO
Venezuelan equine encephalitis virus (VEE) is an important equine and human pathogen of the Americas. In the adult mouse model, cDNA-derived, virulent V3000 inoculated subcutaneously (s.c.) causes high-titer peripheral replication followed by neuroinvasion and lethal encephalitis. A single change (G to A) at nucleotide 3 (nt 3) of the 5' untranslated region (UTR) of the V3000 genome resulted in a virus (V3043) that was avirulent in mice. The mechanism of attenuation by the V3043 mutation was studied in vivo and in vitro. Kinetic studies of virus spread in adult mice following s.c. inoculation showed that V3043 replication was reduced in peripheral organs compared to that of V3000, titers in serum also were lower, and V3043 was cleared more rapidly from the periphery than V3000. Because clearance of V3043 from serum began 1 to 2 days prior to clearance of V3000, we examined the involvement of alpha/beta interferon (IFN-alpha/beta) activity in VEE pathogenesis. In IFN-alpha/betaR(-/-) mice, the course of the wild-type disease was extremely rapid, with all animals dying within 48 h (average survival time of 30 h compared to 7.7 days in the wild-type mice). The mutant V3043 was as virulent as the wild type (100% mortality, average survival time of 30 h). Virus titers in serum, peripheral organs, and the brain were similar in V3000- and V3043-infected IFN-alpha/betaR(-/-) mice at all time points up until the death of the animals. Consistent with the in vivo data, the mutant virus exhibited reduced growth in vitro in several cell types except in cells that lacked a functional IFN-alpha/beta pathway. In cells derived from IFN-alpha/betaR(-/-) mice, the mutant virus showed no growth disadvantage compared to the wild-type virus, suggesting that IFN-alpha/beta plays a major role in the attenuation of V3043 compared to V3000. There were no differences in the induction of IFN-alpha/beta between V3000 and V3043, but the mutant virus was more sensitive than V3000 to the antiviral actions of IFN-alpha/beta in two separate in vitro assays, suggesting that the increased sensitivity to IFN-alpha/beta plays a major role in the in vivo attenuation of V3043.