RESUMO
Spermatogenesis is a highly organized process by which undifferentiated spermatogonia self-renew and differentiate into spermatocytes and spermatids. The entire developmental process from spermatogonia to sperm occurs within the seminiferous tubules. Spermatogenesis is supported by the close interaction of germ cells with Sertoli cells. In this study, testicular tissues were collected from Hu sheep at 8 timepoints after birth: 0, 30, 90, 180, 270, 360, 540, and 720 days. Immunofluorescence staining and histological analysis were used to explore the development of male germ cells and Sertoli cells in the Hu sheep testes at these timepoints. The changes in seminiferous tubule diameter and male germ cells in the Hu sheep testes at these different developmental stages were analyzed. Then, specific molecular markers were used to study the proliferation and differentiation of spermatogonia, the timepoint of spermatocyte appearance, and the maturation and proliferation of Sertoli cells in the seminiferous tubules. Finally, the formation of the blood-testes barrier was studied using antibodies against the main components of the blood-testes barrier, ß-catenin, and ZO-1. These findings not only increased the understanding of the development of the Hu sheep testes, but also laid a solid theoretical foundation for Hu sheep breeding.
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Células de Sertoli , Testículo , Masculino , Animais , Ovinos , Sêmen , Espermatogênese , EspermatogôniasRESUMO
Embryonic stem cells (ESCs) and induced pluripotent stem cells have the potential to differentiate to all cell types of an adult individual and are useful for studying development and for translational research. However, extrapolation of mouse and human ESC knowledge to deriving stable ESC lines of domestic ungulates and large livestock species has been challenging. In contrast to ESCs that are usually established from the blastocyst, mouse expanded potential stem cells (EPSCs) are derived from four-cell and eight-cell embryos. We have recently used the EPSC approach and established stem cells from porcine and human preimplantation embryos. EPSCs are molecularly similar across species and have broader developmental potential to generate embryonic and extraembryonic cell lineages. We further explore the EPSC technology for mammalian species refractory to the standard ESC approaches and report here the successful establishment of bovine EPSCs (bEPSCs) from preimplantation embryos of both wild-type and somatic cell nuclear transfer. bEPSCs express high levels of pluripotency genes, propagate robustly in feeder-free culture, and are genetically stable in long-term culture. bEPSCs have enriched transcriptomic features of early preimplantation embryos and differentiate in vitro to cells of the three somatic germ layers and, in chimeras, contribute to both the embryonic (fetal) and extraembryonic cell lineages. Importantly, precise gene editing is efficiently achieved in bEPSCs, and genetically modified bEPSCs can be used as donors in somatic cell nuclear transfer. bEPSCs therefore hold the potential to substantially advance biotechnology and agriculture.
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Bovinos/genética , Células-Tronco Embrionárias/citologia , Técnicas de Transferência Nuclear/veterinária , Cultura Primária de Células/métodos , Animais , Blastocisto/citologia , Linhagem da Célula , Células Cultivadas , Células-Tronco Embrionárias/metabolismo , Cultura Primária de Células/veterinária , TranscriptomaRESUMO
The mule is the interspecific hybrid of horse and donkey and has hybrid vigor in muscular endurance, disease resistance, and longevity over its parents. Here, we examined adult fibroblasts of mule (MAFs) compared with the cells from their parents (donkey adult fibroblasts and horse adult fibroblasts) (each species has repeated three independent individuals) in proliferation, apoptosis, and glycolysis and found significant differences. We subsequently derived mule, donkey, and horse doxycycline (Dox)-independent induced pluripotent stem cells (miPSCs, diPSCs, and hiPSCs) from three independent individuals of each species and found that the reprogramming efficiency of MAFs was significantly higher than that of cells of donkey and horse. miPSCs, diPSCs, and hiPSCs all expressed the high levels of crucial endogenous pluripotency genes such as POU class 5 homeobox 1 (POU5F1, OCT4), SRY-box 2 (SOX2), and Nanog homeobox (NANOG) and propagated robustly in single-cell passaging. miPSCs exhibited faster proliferation and higher pluripotency and differentiation than diPSCs and hiPSCs, which were reflected in co-cultures and separate-cultures, teratoma formation, and chimera contribution. The establishment of miPSCs provides a unique research material for the investigation of "heterosis" and perhaps is more significant to study hybrid gamete formation.
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Células-Tronco Pluripotentes Induzidas , Cavalos , Animais , Reprogramação Celular , Equidae , Células Cultivadas , Diferenciação Celular/genética , Fibroblastos , Fator 3 de Transcrição de Octâmero/genéticaRESUMO
Many progresses have recently been achieved in animal somatic cell nuclear transfer (SCNT). However, embryos derived from SCNT rarely result in live births. Single-cell RNA sequencing (scRNA-seq) can be used to investigate the development details of SCNT embryos. Here, bovine fibroblasts and three factors bovine iPSCs (3F biPSCs) were used as donors for bovine nuclear transfer, and the single blastomere transcriptome was analysed by scRNA-seq. Compared to in vitro fertilization (IVF) embryos, SCNT embryos exhibited many defects. Abnormally expressed genes were found at each stage of embryos, which enriched in metabolism, and epigenetic modification. The DEGs of the adjacent stage in SCNT embryos did not follow the temporal expression pattern similar to that of IVF embryos. Particularly, SCNT 8-cell stage embryos showed failures in some gene activation, including ZSCAN4, and defects in protein association networks which cored as POLR2K, GRO1, and ANKRD1. Some important signalling pathways also showed incomplete activation at SCNT zygote to morula stage. Interestingly, 3F biPSCNT embryos exhibited more dysregulated genes than SCNT embryos at zygote and 2-cell stage, including genes in KDM family. Pseudotime analysis of 3F biPSCNT embryos showed the different developmental fate from SCNT and IVF embryos. These findings suggested partial reprogrammed 3F biPS cells as donors for bovine nuclear transfer hindered the reprogramming of nuclear transfer embryos. Our studies revealed the abnormal gene expression and pathway activation of SCNT embryos, which could increase our understanding of the development of SCNT embryos and give hints to improve the efficiency of nuclear transfer.
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Clonagem de Organismos , Técnicas de Transferência Nuclear , Animais , Bovinos , Reprogramação Celular/genética , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/genética , Fertilização in vitro , Análise de Sequência de RNA , TranscriptomaRESUMO
BACKGROUND: It is important to resolve the evolutionary history of species genomes as it has affected both genome organization and chromosomal architecture. The rapid innovation in sequencing technologies and the improvement in assembly algorithms have enabled the creation of highly contiguous genomes. DNA Zoo, a global organization dedicated to animal conservation, offers more than 150 chromosome-length genome assemblies. This database has great potential in the comparative genomics field. RESULTS: Using the donkey (Equus asinus asinus, EAS) genome provided by DNA Zoo as an example, the scaffold N50 length and Benchmarking Universal Single-Copy Ortholog score reached 95.5 Mb and 91.6%, respectively. We identified the cytogenetic nomenclature, corrected the direction of the chromosome-length sequence of the donkey genome, analyzed the genome-wide chromosomal rearrangements between the donkey and horse, and illustrated the evolution of the donkey chromosome 1 and horse chromosome 5 in perissodactyls. CONCLUSIONS: The donkey genome provided by DNA Zoo has relatively good continuity and integrity. Sequence-based comparative genomic analyses are useful for chromosome evolution research. Several previously published chromosome painting results can be used to identify the cytogenetic nomenclature and correct the direction of the chromosome-length sequence of new assemblies. Compared with the horse genome, the donkey chromosomes 1, 4, 20, and X have several obvious inversions, consistent with the results of previous studies. A 4.8 Mb inverted structure was first discovered in the donkey chromosome 25 and plains zebra chromosome 11. We speculate that the inverted structure and the tandem fusion of horse chromosome 31 and 4 are common features of non-caballine equids, which supports the correctness of the existing Equus phylogeny to an extent.
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Cromossomos Humanos Par 1 , Equidae , Animais , Cromossomos/genética , Cromossomos Humanos Par 5 , Equidae/genética , Genoma , Cavalos/genética , HumanosRESUMO
The rumen epithelium of sheep serves as an immune interface with the environment and secretes antimicrobial peptides with bactericidal function against various pathogens. Sheep beta-defensin-1 (SBD-1), an antimicrobial peptide, is secreted from ovine ruminal epithelial cells (OREC) in response to microbial infections. Mannan, the main component of the Saccharomyces cerevisiae cell wall can stimulate innate and regulatory immune responses that could improve the gastrointestinal environment. We aimed at investigating the effects of mannan on SBD-1 expression and the downstream signaling pathways stimulated in OREC. We cultured OREC; assessed the effects of mannan on SBD-1 expression by qPCR and ELISA; and then investigated the underlying signaling pathways using qPCR, ELISA, Western blotting, immunohistochemistry, and immunohistofluorescence. Interestingly, mannan markedly upregulated SBD-1 expression in a concentration- and time-dependent manner. Dectin-2 Mouse mAb, Syk specific inhibitor R406, and specific inhibitors of the p38, ERK1/2, JNK, and NF-κB pathways attenuated mannan-induced SBD-1 expression to varying degrees. These results demonstrate that SBD-1 is upregulated by mannan via the Dectin-2-Syk axis, and this is regulated to a large extent through the mitogen-activated protein kinase (MAPK) p38 and less so through the ERK1/2 and JNK or the NF-κB pathway. Our findings highlight the immunomodulatory effects of mannan on OREC in terms of mannan-induced SBD-1 expression.
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Peptídeos Catiônicos Antimicrobianos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Mananas/farmacologia , Carneiro Doméstico/genética , beta-Defensinas/genética , Animais , Células Epiteliais/metabolismo , Lectinas Tipo C/metabolismo , Rúmen/metabolismo , Saccharomyces cerevisiae/química , Carneiro Doméstico/metabolismo , Transdução de Sinais , Quinase Syk/metabolismo , beta-Defensinas/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
To improve the efficiency of somatic cell nuclear transfer (SCNT) in sheep, we investigated the effects of recipient oocyte source, number of transferred embryos and season on the pregnancy and live lamb rates for sheep somatic cell nuclear transfer embryos. Follicle-stimulating hormone (FSH)-stimulated ovaries produced significantly more oocytes both in total and of suitable quality for maturation culture than those without FSH treatment (from slaughterhouse). However, their in vitro maturation rates were similar. Embryos were reconstructed using adult fibroblast cells into enucleated MII oocytes. The pregnancy and term rates were significantly higher in the FSH-stimulated group than in the slaughterhouse one. Oocytes from FSH-stimulated ovaries were enucleated as recipient cytoplasm for nuclear transfer in the following experiments. The transfer of 7-9 and 11-13 embryos produced significantly higher pregnancy rates than that of six embryos. However, the former groups exhibited similar live lamb rates. FSH-stimulated ovaries produced significantly more oocytes in November and December (winter) than in May to July (summer), but the associated maturation rate did not increase. Pregnancy and term rates were significantly higher when transfer occurred in winter than in summer. In conclusion, FSH treatment produced significant benefit regarding the number and quality of collected oocytes and also for the pregnancy and live lamb rates for reconstructed embryos. However, the transfer of an appropriate number of embryos (7-13) and at an appropriate season (winter) increased pregnancy and term rates.
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Transferência Embrionária/veterinária , Técnicas de Transferência Nuclear/veterinária , Estações do Ano , Carneiro Doméstico/fisiologia , Animais , Feminino , Hormônio Foliculoestimulante/farmacologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos , Ovário/efeitos dos fármacos , Gravidez , Taxa de GravidezRESUMO
OBJECTIVE: To explore the relationship between infective endocarditis (IE) and stroke. METHODS: The clinical data of patients diagnosed with IE from January 2003 to December 2017 in Beijing Anzhen Hospital Affiliated to Capital Medical University were retrospectively analyzed. RESULTS: A total of 861 patients (mean age: 40.79 ± 16.27 [SD]) with IE was recruited. Vegetations were confirmed in 97.32% of all the patients, among whom 296 were diagnosed with congenital heart disease and another 53 with rheumatic valvular disease. The most common pathogens were Streptococcus, Staphylococcus, and various types of fungi (13.12, 7.31, and 1.16% respectively). Out of the 138 patients diagnosed with stroke, 101 cases were of ischemic stroke, 23 cases were of hemorrhagic stroke, and 12 cases were of concurrent ischemic and hemorrhagic stroke. There were 31 patients who had infarction lesions in more than 2 vascular systems. The mean age of stroke patients was significantly higher than that of patients without stroke (45.76 ± 17.88 vs. 39.83 ± 15.77, p = 0.000). The incidence of mitral valve vegetation (57.24 vs. 43.01%, p = 0.002), atrial fibrillation (4.34 vs. 1.38%, p = 0.018), fungal infection (2.89 vs. 0.83%, p = 0.038) in patients with stroke was significantly higher than those without stroke. Mitral valve vegetation (OR 1.648; 95% CI 1.113-2.442) and age (OR 1.019; 95% CI 1.007-1.032) were independent risk factors for stroke in IE patients. Stroke increased the risk of hospital deaths (OR 7.673 95%CI 3.634-16.202). CONCLUSION: Stroke is a common complication of IE. Mitral valve vegetation and old age may incerease the risk of stroke in patients with IE.
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Endocardite/complicações , Acidente Vascular Cerebral/etiologia , Adulto , Idoso , Estudos de Coortes , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Acidente Vascular Cerebral/epidemiologiaRESUMO
BACKGROUND: Beta defensins are secreted from ovine oviduct epithelial cells (OOECs) in response to microbial infection, and are potential alternatives to antibiotic agents in the treatment of microorganism infection, particularly given the abuse of antibiotic agents and the increasing number of drug-resistant bacteria. The aberrant expression of defensins may result in disorders involving organ and oviduct inflammation, such as salpingitis. METHODS: In the present study, we investigated the effects of LPS on the mRNA expression levels of sheep ß-defensin-1 (SBD-1) in ovine oviduct epithelial cells. The OOECs in vitro culturing system were established and treated with different concentrations of LPS for indicated time. In addition, MAPK inhibitors and TLR4 antibodies were pretreated to investigate the potential mechanism which involves in LPS regulating SBD-1 expression. RESULTS: LPS markedly upregulated SBD-1 expression in a concentration- and time-dependent manner. Treatment with 100 ng/mL LPS resulted in the phosphorylation of JNK, ERK and P38 MAPK. Interestingly, the LPS stimulated SBD-1 expression was attenuated by pretreatment with the P38 MAPK inhibitors SB203580 and SB202190 but not the JNK inhibitor SP600125, while the ERK inhibitor PD98059 had a minor effect. Furthermore, treatment with a Toll-like receptor 4 (TLR4) neutralizing antibody significantly decreased P38 MAPK phosphorylation and LPS induced SBD-1 expression. CONCLUSIONS: Together, these findings suggest that SBD-1 is upregulated by LPS via the TLR4 receptor, mainly through the P38 MAPK signaling pathway in ovine oviduct epithelial cells to protect the ovine oviduct epithelium from pathogen invasion.
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Tubas Uterinas/metabolismo , Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Ovinos , beta-Defensinas/genética , Animais , Células Cultivadas , Células Epiteliais/metabolismo , Feminino , Fosforilação , RNA Mensageiro/análise , Receptor 4 Toll-Like/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/análise , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologiaRESUMO
Although numerous, human subject studies evaluating the relationship between circulating ghrelin levels and polycystic ovary syndrome (PCOS) risk have yielded inconsistent findings. We aimed to quantitatively assess the association by summarizing all available evidence from human subject studies. The PubMed and Web of Science databases were searched up to February 2015 for eligible studies. Studies were eligible if they reported circulating ghrelin levels in women with PCOS and healthy women controls. A fixed or random-effects model was used to pool risk estimations. Twenty studies including 894 PCOS patients and 574 controls were included in the meta-analysis. The studies had fair methodological quality. The pooling analysis of all available studies revealed that ghrelin levels were significantly lower in PCOS patients than in controls, with standardized mean difference of -0.40 (95% CI: -0.73, -0.08). The significant association persisted in many subgroup strata. However, the heterogeneity across studies was considerable and not eliminated in subgroup analyses. Meta-regression analysis further suggested that the heterogeneity might be relevant to variability in study location, PCOS relevant factors like HOMA-IR ratio, as well as other factors not assessed. In conclusion, our meta-analysis suggested that ghrelin levels were significantly lower in PCOS patients than in controls. Further studies with large sample sizes are warranted to replicate our findings.
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Grelina/sangue , Síndrome do Ovário Policístico/sangue , Adulto , Estudos de Casos e Controles , Regulação para Baixo , Feminino , Humanos , Resistência à InsulinaRESUMO
OBJECTIVE: To investigate the clinical features of stroke in young adults with infective endocarditis (IE). METHODS: This is a retrospective analysis of clinical data of young patients (aged between 18 and 44 years) diagnosed with IE in Beijing Anzhen Hospital affiliated to Capital Medical University from January 2003 to March 2013. RESULTS: Three hundred IE patients (209 male and 91 female) were enrolled with mean age of (31.8 ± 7.8) years old. Fourteen of them were prosthetic valve endocarditis. All of the cases were found vegetations. The vegetations were mainly distributed in the mitral valve and/or aortic valve (262/300 patients). Congenital heart disease and rheumatic valvular disease were most frequently background diseases. Thirty-five cases (11.7%) were complicated with stroke. Among them, 21 cases were with cerebral infarction (including 7 subjects with hemorrhagic transformation), 9 cases with cerebral hemorrhage, 3 cases with subarachnoid hemorrhage, and 2 with reasons unknown. Within the 21 cerebral infarction cases, the infarction sites were mainly distributed in internal carotid artery system with 5 cases involving bilateral internal carotid artery systems and 2 vertebro-basilar artery systems. As to the cerebral hemorrhage, lobar were the bleeding sites in all cases, and mainly distributed in internal carotid artery system. The proportions of left-sided endocarditis and rheumatic valvular disease were significantly higher in patients with stroke than those without [100.0% (35/35) vs 86.4% (229/265), P = 0.02; 22.8% (8/35) vs 6.4% (17/265), P = 0.001, respectively]. Regression analysis showed that the OR of the rheumatic valvular disease for stroke in young IE patients was 4.950 (95% CI 1.626-15.072), and the OR of stroke for death was 8.387 (95% CI 1.997-35.225), respectively. CONCLUSIONS: Stroke is common in young patients with IE. Cerebral infarction often involves multiple vascular systems, and is prone to hemorrhagic transformation. Intracerebral hemorrhage often occurs in lobar. Rheumatic valvular disease might be a risk factor for stroke in young patients with IE, which is an independent risk factor for death of these patients.
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Hemorragia Cerebral/etiologia , Endocardite Bacteriana/complicações , Acidente Vascular Cerebral/etiologia , Adolescente , Adulto , Pequim/epidemiologia , Pesquisa Biomédica , Hemorragia Cerebral/epidemiologia , Infarto Cerebral/epidemiologia , Endocardite Bacteriana/microbiologia , Feminino , Cardiopatias Congênitas/epidemiologia , Humanos , Masculino , Valva Mitral , Estudos Retrospectivos , Cardiopatia Reumática/epidemiologia , Fatores de Risco , Acidente Vascular Cerebral/epidemiologia , Adulto JovemRESUMO
The aim of this study is to explore the mechanism of IL-17A infection in the development of bacterial mastitis in dairy cows. In this study, RT-qPCR and ELISA were used to measure the promoting effect of IL-17A on the generation of pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6) and chemokine (IL-8). In addition, Western blot (WB) was applied to measure the influences of IL-17A on the inflammation-related ERK and p38 proteins in the MAPK pathways. The results show that under the stimulation of LPS on cow mammary epithelial cells (CMECs), cytokines IL-1ß, IL-6, IL-8, TNF-α, and IL-17A will exhibit significantly increased expression levels (p < 0.05). With inhibited endogenous expression of IL-17A, cytokines IL-1ß, IL-6, IL-8, and TNF-α will present reduced genetic expression (p < 0.01), with reduced phosphorylation levels of ERK and p38 proteins in the MAPK signaling pathways (p < 0.001). Upon the exogenous addition of the IL-17A cytokine, the genetic expression of cytokines IL-1ß, IL-6, IL-8, and TNF-α will increase (p < 0.05), with increased phosphorylation levels of the ERK and p38 proteins in the MAPK signaling pathways (p < 0.001). These results indicate that under the stimulation of CMECs with LPS, IL-17A can be expressed together with relevant inflammatory cytokines. Meanwhile, the inflammatory responses of mammary epithelial cells are directly proportional to the expression levels of IL-17A inhibited alone or exogenously added. In summary, this study shows that IL-17A could be used as an important indicator for assessing the bacterial infections of mammary glands, indicating that IL-17A could be regarded as one potential therapeutic target for mastitis.
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With the change in diet structure, individuals prefer to consume mutton with less fat. However, sheep tail has a lot of fat. We identified a breed of low-fat short-tailed sheep (i.e., Hulunbuir short-tailed sheep). It is necessary to develop an animal model that can promote research on the potential mechanisms of the short-tail phenotype in sheep, which results from the TBXT gene c.G334T mutation. To create animal models, we selected mice as experimental animals. Mouse embryos lacking the TBXT protein, which crucially regulates mouse embryonic development, cannot develop normally. We utilized CRISPR/Cas9 gene editing technology to generate site-specific mutation (c.G334T) in the TBXT gene of mice, and found that the mouse TBXT mutation (c.G334T) leads to a short-tail phenotype. Furthermore, we investigated the interaction between TBXT and Wnt signaling pathways. The expressions of TBXT, Axin2, Dkk1, Wnt3, Wnt3a, and Wnt5a were discovered to be significantly different between mutant embryos and wild embryos by obtaining mouse embryos at various developmental stages and examining the expression relationship between the TBXT and Wnt signaling pathway-related components in all of these embryos. Therefore, as a transcription factor, TBXT regulates the expression of the aforementioned Wnt signaling pathway components by forming a regulatory network for the normal development of mouse embryos. This study enriches the research on the functional role of the TBXT in the development of mouse embryos and the mechanism by which the short-tailed phenotype in sheep develops.
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Sistemas CRISPR-Cas , Cauda , Gravidez , Feminino , Camundongos , Animais , Ovinos/genética , Desenvolvimento Embrionário/genética , Fenótipo , Edição de Genes/métodosRESUMO
A minority of mouse embryonic stem cells (ESCs) display totipotent features resembling 2-cell stage embryos and are known as 2-cell-like (2C-like) cells. However, how ESCs transit into this 2C-like state remains largely unknown. Here, we report that the overexpression of negative elongation factor A (Nelfa), a maternally provided factor, enhances the conversion of ESCs into 2C-like cells in chemically defined conditions, while the deletion of endogenous Nelfa does not block this transition. We also demonstrate that Nelfa overexpression significantly enhances somatic cell reprogramming efficiency. Interestingly, we found that the co-overexpression of Nelfa and Bcl2 robustly activates the 2C-like state in ESCs and endows the cells with dual cell fate potential. We further demonstrate that Bcl2 overexpression upregulates endogenous Nelfa expression and can induce the 2C-like state in ESCs even in the absence of Nelfa. Our findings highlight the importance of BCL2 in the regulation of the 2C-like state and provide insights into the mechanism underlying the roles of Nelfa and Bcl2 in the establishment and regulation of the totipotent state in mouse ESCs.
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Células-Tronco Embrionárias , Células-Tronco Embrionárias Murinas , Animais , Camundongos , Células-Tronco Embrionárias Murinas/metabolismo , Diferenciação Celular , Células-Tronco Embrionárias/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
The significant deposition of tail fat in sheep has a profound impact on the economic benefits of animal husbandry. Furthermore, increasing the litter size is a crucial means of enhancing economic benefits. The BMPR1B and T/Brachyury genes are considered major functional genes that could affect sheep litter size and tail bone number, respectively. In this study, we employed direct sequencing to identify specific mutations of the BMPR1B gene in Gobi short tail sheep and carried out genotyping using MassARRAY technology for each variant of both the BMPR1B and T genes. Significant associations were demonstrated between the c.687G>A mutation of BMPR1B and the litter size in both the Gobi short tail sheep and Ujimqin sheep breeds. Meanwhile, the g.30058882_30058873GCAGATTAAAIndel mutation was significantly associated with the litter size in Gobi short tail sheep. These findings may provide valuable genetic markers for expanding sheep litter size. In addition, we also confirmed that the frequency of tail-bone-number-related T alleles was significantly higher in Gobi short tail sheep than in longer-tailed Ujimqin sheep.
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The number of vertebrae is a crucial economic trait that can significantly impact the carcass length and meat production in animals. However, our understanding of the quantitative trait loci (QTLs) and candidate genes associated with the vertebral number in sheep (Ovis aries) remains limited. To identify these candidate genes and QTLs, we collected 73 Ujimqin sheep with increased numbers of vertebrae (T13L7, T14L6, and T14L7) and 23 sheep with normal numbers of vertebrae (T13L6). Through high-throughput genome resequencing, we obtained a total of 24,130,801 effective single-nucleotide polymorphisms (SNPs). By conducting a selective-sweep analysis, we discovered that the most significantly selective region was located on chromosome 7. Within this region, we identified several genes, including VRTN, SYNDIG1L, LTBP2, and ABCD4, known to regulate the spinal development and morphology. Further, a genome-wide association study (GWAS) performed on sheep with increased and normal vertebral numbers confirmed that ABCD4 is a candidate gene for determining the number of vertebrae in sheep. Additionally, the most significant SNP on chromosome 7 was identified as a candidate QTL. Moreover, we detected two missense mutations in the ABCD4 gene; one of these mutations (Chr7: 89393414, C > T) at position 22 leads to the conversion of arginine (Arg) to glutamine (Gln), which is expected to negatively affect the protein's function. Notably, a transcriptome expression profile in mouse embryonic development revealed that ABCD4 is highly expressed during the critical period of vertebral formation (4.5-7.5 days). Our study highlights ABCD4 as a potential major gene influencing the number of vertebrae in Ujimqin sheep, with promising prospects for future genome-assisted breeding improvements in sheep.
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OBJECTIVE: To summarize the characteristics of neurological symptoms in patients with aortic dissection. METHODS: The clinical data including symptoms, signs, and imaging of 865 consecutive patients with aortic dissection were analyzed retrospectively. RESULTS: Neurological symptoms occurred in 225 cases (26.0%) with aortic dissection. The most common symptom was dizziness (56 cases, 6.5%), followed by syncope (49, 5.7%), single lower limb sensory disturbances (47, 5.4%), single lower extremity weakness (27, 3.1%), coma (22, 2.5%), paraplegia (19, 2.2%), headache (13,1.5%) and hemiplegia (9,1.0%). The first manifestation of five cases was the neurological symptoms, including syncope (2), dizziness (1) and headache (2). Patients with type A aortic dissection were more vulnerable to the neurological symptoms than those with type B aortic dissection (34.6% vs 14.7%), and the symptoms with significantly higher occurrence were syncope, coma, hemiplegia, paraplegia and lower limb sensory disturbances. CONCLUSION: Neurological symptoms are common in patients with aortic dissection,especially in those with type A aortic dissection.
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Aneurisma Aórtico/diagnóstico , Dissecção Aórtica/diagnóstico , Doenças do Sistema Nervoso/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doenças do Sistema Nervoso/patologia , Estudos Retrospectivos , Adulto JovemRESUMO
This study was designed to investigate the promotion of oxidation of lipids in oil-in-water (o/w) emulsions and digestive properties of the bionic dynamic gastrointestinal system of whey protein isolate (WPI) and gum arabic (GA) complexes loaded with iron ions, which were fabricated previously and shown as WPI/GAFe3+ nanoparticles (WGS) and WPI/GAFe3+ fibers (WGF). Compared with emulsions containing Fe3+ and GA-loaded complex (GAFe3+), WGS and WGF greatly improved the oxidative stability of lipids along with the reduced lipid oxidation products and volatile compounds, attributed to the encapsulation of iron ions. During the bionic dynamic gastrointestinal digestion, the iron ion release of WGF was significantly higher than that of WGS, probably due to different assembled internal structures. Accordingly, two proposed WPI/GAFe3+ complexes with different morphologies are expected to be developed as novel stable iron fortifiers with delayed lipid oxidation and controlled iron-ion release in food emulsions.
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Goma Arábica , Ferro , Goma Arábica/química , Proteínas do Soro do Leite/química , Ferro/química , Emulsões/química , Lipídeos , Água/químicaRESUMO
Intramuscular fat content and marbling affecting meat quality are important economic traits in beef cattle. CDC10 (cell division cycle 10 or Septin 7), a member of the septin family involved in cellular proliferation, was considered as a functional and positional candidate gene for beef marbling. In a previous study, we revealed that the expression levels of CDC10 were also positively correlated with marbling scores in Japanese Black cattle. However, the regulatory mechanism of the CDC10 gene on IMF deposition in cattle remains unclear. In the present study, flow cytometry, EdU proliferation assays, and Oil Red O staining results showed that overexpression of CDC10 could promote the differentiation of bovine intramuscular preadipocyte (BIMP) and 3T3-L1 cells, whereas knockdown of CDC10 resulted in the opposite consequences. Furthermore, quantitative PCR and Western blotting results showed that overexpression of CDC10 could promote the expression levels of adipogenic marker genes PPARγ and C/EBPα at both mRNA and protein levels in BIMP and 3T3-L1 cells, whereas knockdown of CDC10 resulted in the opposite consequences. Our results provide new insights into the regulatory roles of CDC10 in adipocytes in animals.
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The process of testis development in mammals is accompanied by the proliferation and maturation of Sertoli, Leydig and germ cells. Spermatogenesis depends on hormone regulation, which must bind to a receptor to exert its biological effects. The changes in Hu sheep testis cell composition and FSHR, LHR and AR expression during different developmental stages are unclear (newborn, puberty and adulthood). To address this, using single-cell RNA sequencing, we analyzed testis cell composition and hormone receptor expression changes during three important developmental stages of Hu sheep. We observed significant changes in the composition of somatic and germ cells in different Hu sheep testis developmental stages. Furthermore, we analyzed the FSHR, LHR and AR distribution and expression changes at three important periods and verified them by qRT-PCR and immunofluorescence. Our results suggest that after birth, the proportion of germ cells increased gradually, peaking in adulthood; the proportion of Sertoli cells decreased gradually, reaching the lowest in adulthood; and the proportion of Leydig cells increased and then decreased, reaching the lowest in adulthood. In addition, FSHR, LHR and AR are mainly located in Sertoli, Leydig and germ cells. LHR and FSHR expression decreased with increasing age, while AR expression increased and then decreased with increasing age.