RESUMO
In pig production, pigs often show more aggressive behavior after mixing, which adversely affects animal welfare and growth performance. The Jumonji and structural domain-rich AT interaction domain 2 (JARID2) gene plays an important role in neurodevelopment in mice and various psychiatric disorders in humans. The JARID2 gene may impact the aggressive behavior of pigs. By observing the behavior of 500 weaned pigs during the first 72 h after mixing, the ear tissue samples of the 12 most aggressive and 12 least aggressive pigs were selected for DNA resequencing based on the intensity of their aggressive behavior. Large group correlation analysis indicated that the rs3262221458 site located in the 3'-UTR region of the porcine JARID2 gene has a strong relationship with the aggressive behavior of weaned pigs. Pigs with the mutant TT genotype of rs3262221458 have more aggressive behavior than those pigs with the GG and GT genotypes. The dual luciferase assay indicated that the luciferase activity of the plasmids containing the G allele of rs326221458 was significantly less than that of plasmids containing the T allele of rs326221458 and control groups. The binding ability of miR-9828-3p to sequences containing the T allele was less than that of sequences containing the G allele. The overexpression of miR-9828-3p in porcine neuroglial cells (PNGCs) and PK15 cells significantly decreased the mRNA and protein levels of the JARID2 gene. In addition, miR-9828-3p inhibited the proliferation of PNGCs. After inhibiting miR-9828-3p, the mRNA and protein expression levels of JARID2 increased, and the proliferation of PNGCs showed an opposite trend to the cells that forced the expression of miR-9828-3p. In addition, interference with the JARID2 gene by siRNA can effectively inhibit the proliferation of PNGCs. In summary, we found that the rs326221458 locus regulates the expression of the JARID2 gene by affecting the binding of miR-9828-3p and the JARID2 gene, thereby affecting the aggressive behavior of weaned pigs after mixing.
Assuntos
MicroRNAs , Polimorfismo de Nucleotídeo Único , Humanos , Suínos/genética , Animais , Camundongos , Regiões 3' não Traduzidas , RNA Mensageiro , Luciferases , MicroRNAs/genética , Complexo Repressor Polycomb 2RESUMO
OBJECTIVE: This study explores the willingness of older adults to use smartphones and improve their digital skills and encourages nursing to actively participate in bridging the digital divide. METHODS: Subject analysis was used to conduct qualitative research, and 23 older adults were interviewed. RESULTS: We identified four themes: (1) the current situation of smartphone use; (2) the digital dilemma of smartphone use; (3) social support for digital skills; and (4) the willingness to learn digital skills. Older adults in China are willing to accept and use smartphones for simple operations, and peer learning may be an effective way to improve their digital skills. CONCLUSION: Community support is necessary to develop the digital skills of older adults with smartphones and reduce the digital divide to the greatest extent possible. Nursing may play a role in promoting digital inclusion for older adults.
Assuntos
População do Leste Asiático , Smartphone , Humanos , Idoso , Aprendizagem , Apoio Social , Pesquisa QualitativaRESUMO
PURPOSE: To examine key impacts of anesthesia on new-onset atrial fibrillation (AF) and acute kidney injury (AKI) in transcatheter aortic valve replacement (TAVR). METHODS: All consecutive patients who underwent transfemoral, transapical, and transaortic TAVR in Fuwai Hospital from 2012 to 2018 were retrospectively analyzed and dichotomized into 2 groups: TAVR under conscious sedation (CS) and under general anesthesia (GA). The primary endpoint was a composite of all-cause mortality, stroke, AF, permanent pacemaker implantation, myocardial infarction, heart failure, high-grade atrioventricular block, and AKI at 1 year. Binary logistic regression and adjusted multilevel logistic regression were performed to analyze the predictors of AF and AKI. RESULTS: A total of 107 patients were under CS and 66 patients under GA. No significant difference was observed in the composite endpoint (51.5% vs. 41.2%, GA vs. CS, P = .182) and ≥ mild paravalvular leakage (36.4% vs. 31.4%, GA vs. CS, P = .589) at 1 year. However, the GA group had a significantly higher rate of intensive care unit (ICU) admission (84.8% vs. 6.5%, P < .001), AKI (28.8% vs. 14.0%, P = .018), new-onset AF (15.2% vs. 5.5% at 1 year, P = .036). Multivariable analysis revealed GA to be the significant predictor of new-onset AF (odds ratio 3.237, 95% confidence interval 1.059 to 9.894, P = .039) and AKI (odds ratio 2.517, 95% confidence interval 1.013 to 6.250, P = .047). CONCLUSION: GA was associated with higher rates of ICU admission, postoperative AKI, and new-onset AF. The results may provide new evidence that CS challenges universal GA.
Assuntos
Injúria Renal Aguda/etiologia , Anestesia Geral/efeitos adversos , Estenose da Valva Aórtica/cirurgia , Fibrilação Atrial/etiologia , Complicações Pós-Operatórias , Medição de Risco/métodos , Substituição da Valva Aórtica Transcateter/métodos , Injúria Renal Aguda/epidemiologia , Idoso , Valva Aórtica/cirurgia , Estenose da Valva Aórtica/mortalidade , Fibrilação Atrial/epidemiologia , China/epidemiologia , Feminino , Seguimentos , Mortalidade Hospitalar/tendências , Humanos , Incidência , Masculino , Tomografia Computadorizada Multidetectores/métodos , Estudos Retrospectivos , Fatores de Risco , Taxa de Sobrevida/tendênciasRESUMO
BACKGROUND: Levosimendan has been shown to confer direct renoprotection in renal endotoxemic and ischemia-reperfusion injury and could increase renal blood flow in patients with low-cardiac-output heart failure. Results from clinical trials of levosimendan on acute kidney injury (AKI) following cardiac surgery are controversial. STUDY DESIGN: A random-effect meta-analysis was conducted based on evidence from PubMed, EMBASE, and Cochrane Library. SETTINGS & POPULATION: Adult patients undergoing cardiac surgery. SELECTION CRITERIA FOR STUDIES: Randomized controlled trials comparing the renal effect of levosimendan versus placebo or other inotropic drugs during cardiac surgery. INTERVENTION: Perioperative levosimendan continuous infusion at a rate of 0.1 to 0.2µg/kg/min following a loading dose (6-24µg/kg) for 24 hours or only 1 loading dose (24µg/kg) within 1 hour. OUTCOMES: AKI, need for renal replacement therapy, mechanical ventilation duration, intensive care unit stay during hospitalization, and postoperative mortality (in-hospital or within 30 days). RESULTS: 13 trials with a total of 1,345 study patients were selected. Compared with controls, levosimendan reduced the incidence of postoperative AKI (40/460 vs 78/499; OR, 0.51; 95% CI, 0.34-0.76; P=0.001; I(2)=0.0%), renal replacement therapy (22/492 vs 49/491; OR, 0.43; 95% CI, 0.25-0.76; P=0.002; I(2)=0.0%), postoperative mortality (35/658 vs 94/657; OR, 0.41; 95% CI, 0.27-0.62; P<0.001; I(2)=0.0%), mechanical ventilation duration (in days; n=235; weighted mean difference, -0.34; 95% CI, -0.58 to -0.09; P=0.007], and intensive care unit stay (in days; n=500; weighted mean difference, -2.2; 95% CI, -4.21 to -0.13; P=0.04). LIMITATIONS: Different definitions for AKI among studies. Small sample size for some trials. CONCLUSIONS: Perioperative administration of levosimendan in patients undergoing cardiac surgery may reduce complications. Future trials are needed to determine the dose effect of levosimendan in improving outcomes, especially in patients with decreased baseline kidney function.
Assuntos
Injúria Renal Aguda , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Hidrazonas/farmacologia , Rim/efeitos dos fármacos , Complicações Pós-Operatórias/prevenção & controle , Piridazinas/farmacologia , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/prevenção & controle , Adulto , Procedimentos Cirúrgicos Cardíacos/métodos , Humanos , Avaliação de Resultados em Cuidados de Saúde , Substâncias Protetoras/farmacologia , Ensaios Clínicos Controlados Aleatórios como Assunto , Simendana , Vasodilatadores/farmacologiaRESUMO
Pork belly, prized for its unique flavor and texture, is often overlooked in breeding programs that prioritize lean meat production. The quality of pork belly is determined by the number and distribution of muscle and fat layers. This study aimed to assess the number of pork belly layers using deep learning techniques. Initially, semantic segmentation was considered, but the intersection over union (IoU) scores for the segmented parts were below 70%, which is insufficient for practical application. Consequently, the focus shifted to image classification methods. Based on the number of fat and muscle layers, a dataset was categorized into three groups: three layers (n = 1811), five layers (n = 1294), and seven layers (n = 879). Drawing upon established model architectures, the initial model was refined for the task of learning and predicting layer traits from B-ultrasound images of pork belly. After a thorough evaluation of various performance metrics, the ResNet18 model emerged as the most effective, achieving a remarkable training set accuracy of 99.99% and a validation set accuracy of 96.22%, with corresponding loss values of 0.1478 and 0.1976. The robustness of the model was confirmed through three interpretable analysis methods, including grad-CAM, ensuring its reliability. Furthermore, the model was successfully deployed in a local setting to process B-ultrasound video frames in real time, consistently identifying the pork belly layer count with a confidence level exceeding 70%. By employing a scoring system with 100 points as the threshold, the number of pork belly layers in vivo was categorized into superior and inferior grades. This innovative system offers immediate decision-making support for breeding determinations and presents a highly efficient and precise method for assessment of pork belly layers.
RESUMO
A comprehensive understanding of the complex regulatory mechanisms governing estrus and ovulation across multiple tissues in mammals is imperative to improve the reproductive performance of livestock and mitigate ovulation-related disorders in humans. To comprehensively elucidate the regulatory landscape, we analyzed the transcriptome of protein-coding genes and long intergenic non-coding RNAs (lincRNAs) in 58 samples (including the hypothalamus, pituitary, ovary, vagina, and vulva) derived from European Large White gilts and Chinese Mi gilts during estrus and diestrus. We constructed an intricate regulatory network encompassing 358 hub genes across the five examined tissues. Furthermore, our investigation identified 85 differentially expressed lincRNAs that are predicted to target 230 genes associated with critical functions including behavior, receptors, and apoptosis. Importantly, we found that vital components of estrus and ovulation events involve "Apoptosis" pathway in the hypothalamus, "Autophagy" in the ovary, as well as "Hypoxia" and "Angiogenesis" in the vagina and vulva. We have identified several differentially expressed transcription factors (TFs), such as SPI1 and HES2, which regulate these pathways. SPI1 may suppress transcription in the autophagy pathway, promoting apoptosis and inhibiting the proliferation of ovarian granulosa cells. Our study provides the most comprehensive transcriptional profiling information related to estrus and ovulation events.
Assuntos
RNA Longo não Codificante , Transcriptoma , Humanos , Suínos , Feminino , Animais , Transcriptoma/genética , Diestro/fisiologia , Estro/genética , Sus scrofa/genéticaRESUMO
The fertility rate and litter size of female pigs are critically affected by the expression of estrus. The objective of this study was to elucidate the regulatory mechanisms of estrus expression by analyzing the differential expression of genes and long intergenic non-coding RNAs (lincRNA), as well as the utilization of alternative polyadenylation (APA) sites, in the vulva and vagina during the estrus and diestrus stages of Large White and indigenous Chinese Mi gilts. Our study revealed that the number of differentially expressed genes (DEG) in the vulva was less than that in the vagina, and the DEGs in the vulva were enriched in pathways such as "neural" pathways and steroid hormone responses, including the "Calcium signaling pathway" and "Oxytocin signaling pathway". The DEGs in the vagina were enriched in the "Metabolic pathways" and "VEGF signaling pathway". Furthermore, 27 and 21 differentially expressed lincRNAs (DEL), whose target genes were enriched in the "Endocrine resistance" pathway, were identified in the vulva and vagina, respectively. Additionally, we observed that 63 and 618 transcripts of the 3'-untranslated region (3'-UTR) were lengthened during estrus in the vulva and vagina, respectively. Interestingly, the genes undergoing APA events in the vulva exhibited species-specific enrichment in neural or steroid-related pathways, whereas those in the vagina were enriched in apoptosis or autophagy-related pathways. Further bioinformatic analysis of these lengthened 3'-UTRs revealed the presence of multiple miRNAs binding sites and cytoplasmic polyadenylation element (CPE) regulatory aspects. In particular, we identified more than 10 CPEs in the validated lengthened 3'-UTRs of the NFIX, PCNX4, CEP162 and ABHD2 genes using RT-qPCR. These findings demonstrated the involvement of APA and lincRNAs in the regulation of estrus expression in female pigs, providing new insights into the molecular mechanisms underlying estrus expression in pigs.
RESUMO
BACKGROUND: Trigeminal neuralgia is a common chronic maxillofacial neuropathic pain disorder, and voltage-gated sodium channels (VSGCs) are likely involved in its pathology. Prior studies report that pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide highly expressed in the trigeminal ganglion, may contribute to dorsal root ganglion neuron excitability by modulating the Nav1.7. OBJECTIVE: We investigated whether PACAP can regulate Nav1.7 through the mitogen-activated protein kinase/ERK kinase/extracellular-signal-regulated kinase (MEK/ERK) pathway in the trigeminal ganglion after chronic constriction injury of the infraorbital nerve (ION-CCI) in rats. STUDY DESIGN: Sprague-Dawley rats underwent ION-CCI, followed by intrathecal injection of PACAP 6-38 (PAC1 receptor antagonist) and PD98059 (MEK/ERK antagonist). Quantitative real-time PCR and western blot were used to quantify ATF3, PACAP, ERK, p-ERK, and Nav1.7 expression. RESULTS: The mechanical pain threshold decreased from day 3 to day 21 after ION-CCI and reached the lowest testing value by day 14; however, it increased after PACAP 6-38 and PD98059 injections. Additionally, ION-CCI surgery increased ATF3, PACAP, and p-ERK expression in the rat trigeminal ganglion and decreased Nav1.7 and PAC1 receptor expression; however, there was no difference in ERK expression. PACAP 6-38 injection significantly decreased PACAP, p-ERK, and Nav1.7 expression and increased the PAC1 receptor expression, with no change in ERK expression. Moreover, PD98059 injection decreased PACAP, p-ERK, and Nav1.7 expression and increased the expression of PAC1 receptor. CONCLUSION: After ION-CCI, PACAP in the rat trigeminal ganglion can modulate Nav1.7 through the MEK/ERK pathway via the PAC1 receptor. Further, PACAP inhibition alleviates allodynia in ION-CCI rats.
Assuntos
Sistema de Sinalização das MAP Quinases , Neuralgia , Ratos , Animais , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Ratos Sprague-Dawley , Receptores de Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Constrição , Neuralgia/tratamento farmacológico , Transdução de Sinais , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismoRESUMO
Copy number variation (CNV) represents a significant reservoir of genetic diversity within the genome and exhibits a strong association with economically valuable traits in livestock. The manifestation of aggressive behavior in pigs has detrimental effects on production efficiency, immune competency, and meat quality. Nevertheless, the impact of CNV on the aggressive behavior of pigs remains elusive. In this investigation, we employed an integrated analysis of genome and transcriptome data to investigate the interplay between CNV, gene expression changes, and indicators of aggressive behavior in weaned pigs. Specifically, a subset of pigs comprising the most aggressive pigs (MAP, n = 12) and the least aggressive pigs (LAP, n = 11) was purposefully selected from a herd of 500 weaned pigs following a mixing procedure based on their composite aggressive score (CAS). Subsequently, we thoroughly analyzed copy number variation regions (CNVRs) across the entire genome using next-generation sequencing techniques, ultimately revealing the presence of 6869 CNVRs. Using genome-wide association study (GWAS) analysis and evaluating variance-stabilizing transformation (VST) values, we successfully identified distinct CNVRs that distinguished the MAP and LAP counterparts. Among the prioritized CNVRs, CNVR-4962 (designated as the top-ranked p-value and VST value, No. 1) was located within the Solute Carrier Organic Anion Transporter Family Member 3A1 (SLCO3A1) gene. The results of our analyses indicated a significantly higher (p < 0.05) copy number of SLCO3A1 in the MAP compared to the LAP. Furthermore, this increased copy number exhibited a positive correlation with the CAS of the pigs (p < 0.05). Furthermore, we integrated genomic data with transcriptomic data from the temporal lobe to facilitate the examination of expression quantitative trait loci (eQTL). Importantly, these observations were consistent with the mRNA expression pattern of SLCO3A1 in the temporal lobe of both MAP and LAP (p < 0.05). Consequently, our findings strongly suggest that CNVs affecting SLCO3A1 may influence gene expression through a dosage effect. These results highlight the potential of SLCO3A1 as a candidate gene associated with aggressive traits in pig breeding programs.
Assuntos
Agressão , Variações do Número de Cópias de DNA , Estudo de Associação Genômica Ampla , Transportadores de Ânions Orgânicos , Animais , Variações do Número de Cópias de DNA/genética , Perfilação da Expressão Gênica , Genômica , Suínos/genética , Transcriptoma , Transportadores de Ânions Orgânicos/genética , Agressão/fisiologiaRESUMO
As a natural sweetener, stevioside is extracted from Stevia rebaudiana Bertoni and possesses potent antioxidant activity. However, little information is known about its protective role in maintaining the intestinal epithelial cells health under oxidative stress. The aim of this study was to investigate the protective effects and underlying mechanisms of stevioside on alleviating inflammation, apoptosis, and improving antioxidant capacity in intestinal porcine epithelial cells (IPEC-J2) under oxidative stress by diquat. The results demonstrated that the pretreatment with stevioside (250 µM) for 6 h increased cell viability and proliferation and prevented apoptosis induced by diquat at 1000 µM for 6 h in IPEC-J2 cells, compared with the diquat alone-treated cells. Importantly, stevioside pretreatment significantly reduced ROS and MDA production as well as upregulated T-SOD, CAT, and GSH-Px activity. Moreover, it also decreased cell permeability and improved intestinal barrier functions by significantly upregulating the tight junction protein abundances of claudin-1, occludin, and ZO-1. At the same time, stevioside significantly down-regulated the secretion and gene expression of IL-6, IL-8, and TNF-α and decreased the phosphorylation levels of NF-κB, IκB, and ERK1/2 compared with the diquat alone group. Taken together, this study demonstrated that stevioside alleviated diquat-stimulated cytotoxicity, inflammation, and apoptosis in IPEC-J2 cells, protecting cellular barrier integrity and mitigating oxidative stress by interfering with the NF-κB and MAPK signaling pathways.
RESUMO
BACKGROUND: Bleeding and the need for allogeneic transfusions are still problems after off-pump coronary artery bypass grafting (OPCAB) surgery. We therefore evaluated the effects of an antifibrinolytic, tranexamic acid, on postoperative bleeding and transfusion requirements in patients undergoing OPCAB surgery. METHODS: Two hundred thirty-one consecutive patients scheduled for elective OPCAB were enrolled in the study. Using a double-blind method, the patients were randomly assigned to receive either tranexamic acid (bolus 1 g before surgical incision followed by an infusion of 400 mg/h during surgery; n = 116) or a placebo (infusion equivalent volume of saline solution; n = 115). The primary outcome was 24-hour postoperative chest tube drainage. Allogeneic transfusion, mortality, major morbidities, and resource utilization were also recorded. RESULTS: In comparison with the placebo group, the patients receiving tranexamic acid had a significant reduction in chest tube drainage at 6 hours (270 ± 118 mL vs 416 ± 179 mL, P < 0.001) and 24 hours (654 ± 224 mL vs 891 ± 295 mL, P < 0.001). There was also a significant reduction in allogeneic red blood cell transfusions (47 vs 31.9%, P = 0.019) and fresh frozen plasma (29.6% vs 17.2%, P = 0.027) transfusions. There were no differences in mortality, morbidity, and resource utilization between the 2 groups. CONCLUSIONS: Tranexamic acid reduces postoperative chest tube drainage and the requirement for allogeneic transfusion in off-pump coronary surgery.
Assuntos
Antifibrinolíticos/uso terapêutico , Ponte de Artéria Coronária sem Circulação Extracorpórea/efeitos adversos , Doença da Artéria Coronariana/cirurgia , Hemorragia Pós-Operatória/prevenção & controle , Ácido Tranexâmico/uso terapêutico , Idoso , Antifibrinolíticos/efeitos adversos , Tubos Torácicos , Distribuição de Qui-Quadrado , China , Ponte de Artéria Coronária sem Circulação Extracorpórea/mortalidade , Doença da Artéria Coronariana/mortalidade , Método Duplo-Cego , Drenagem/instrumentação , Procedimentos Cirúrgicos Eletivos , Transfusão de Eritrócitos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Placebos , Hemorragia Pós-Operatória/etiologia , Hemorragia Pós-Operatória/mortalidade , Estudos Prospectivos , Medição de Risco , Fatores de Risco , Fatores de Tempo , Ácido Tranexâmico/efeitos adversos , Resultado do TratamentoRESUMO
The cooperation between emerging enterprises and different enterprises can stimulate innovation enthusiasm and realize synergistic value creation. At present, there is no regular pattern and rules for synergistic value creation among emerging enterprises, which cannot achieve effective synergistic value creation. Based on the complex network relationship and dynamic model of game evolution among multiple entities of value creation within emerging enterprises, the factors that affect the efficiency of synergistic value creation in the aspect of input, benefit assignment, and interaction mechanism of synergistic value creation are analyzed and then the path of the promotion of synergistic value creation among the entities of new enterprises is studied. The results show that the core enterprises play a leading role in the cooperation input among emerging enterprises, and if the balance of interests among cooperative enterprises is guaranteed, the revenue effect is the best, and the network cooperation density is the strongest. The game evolution shows that, in order to improve the efficiency of synergistic value creation, it is necessary to improve the expected coefficient and number of synergies between enterprises and establish a mechanism for equitable distribution of synergies by strengthening information exchange among enterprises in emerging industries, so as to build an atmosphere of synergies, the complementarity of assets and synergies.
Assuntos
Indústrias , ChinaRESUMO
In this study, we evaluated the protective effects and potential mechanisms of acidifiers on intestinal epithelial cells exposure to oxidative stress (OS). IPEC-J2 cells were first pretreated with 5 × 10-5 acidifiers for 4 h before being exposed to the optimal dose of diquat to induce oxidative stress. The results showed that acidifiers attenuated diquat-induced oxidative stress, which manifests as the improvement of antioxidant capacity and the reduction in reactive oxygen species (ROS) accumulation. The acidifier treatment decreased cell permeability and enhanced intestinal epithelial barrier function through enhancing the expression of claudin-1 and occludin in diquat-induced cells. Moreover, acidifier treatment attenuated diquat-induced inflammatory responses, which was confirmed by the decreased secretion and gene expression of pro-inflammatory (TNF-α, IL-8) and upregulated anti-inflammatory factors (IL-10). In addition, acidifiers significantly reduced the diquat-induced gene and protein expression levels of COX-2, NF-κB, I-κB-ß, ERK1/2, and JNK2, while they increased I-κB-α expression in IPEC-J2 cells. Furthermore, we discovered that acidifiers promoted epithelial cell proliferation (increased expression of PCNA and CCND1) and inhibited apoptosis (decreased expression of BAX, increased expression of BCL-2). Taken together, these results suggest that acidifiers are potent antioxidants that attenuate diquat-induced inflammation, apoptosis, and maintain cellular barrier integrity by regulating the NF-κB/MAPK/COX-2 signaling pathways.
RESUMO
Pigs are important biomedical model animals for the study of human neurological diseases. Similar to human aggressive behavior in children and adolescents, weaned pigs also show more aggressive behavior after mixing, which has negative effects on animal welfare and growth performance. The identification of functional single-nucleotide polymorphisms (SNPs) related to the aggressive behavior of pigs would provide valuable molecular markers of the aggressive behavioral trait for genetic improvement program. The Rho GTPase-activating protein 24 (ARHGAP24) gene plays an important role in regulating the process of axon guidance, which may impact the aggressive behavior of pigs. By resequencing the entire coding region, partially adjacent introns and the 5' and 3' flanking regions, six and four SNPs were identified in the 5' flanking region and 5' untranslated region (UTR) of the porcine ARHGAP24 gene, respectively. Association analyses revealed that nine SNPs were significantly associated with aggressive behavioral traits (p = < 1.00 × 10-4-4.51 × 10-2), and their haplotypes were significantly associated with aggressive behavior (p = < 1.00 × 10-4-2.99 × 10-2). The core promoter region of the ARHGAP24 gene has been identified between -670 and -1,113 bp. Furthermore, the luciferase activity of allele A of rs335052970 was significantly less than that of allele G, suggesting that the transcriptional activity of the ARHGAP24 gene was inhibited by allele A of rs335052970. It was identified that the transcription factor p53 bound to the transcription factor binding sites (TFBSs) containing allele A of rs335052970. In porcine primary neural cells, p53 binds to the target promoter region of the ARHGAP24 gene, reduces its promoter transcriptional activity, and then reduces its messenger RNA (mRNA) and protein expression. The results demonstrated that the ARHGAP24 gene had significant genetic effects on aggressive behavioral traits of pigs. Therefore, rs335052970 in the ARHGAP24 gene can be used as a molecular marker to select the less aggressive pigs.
RESUMO
Chinese indigenous Mi gilts have clearer estrus expression than European Large White gilts, and sulfotransferase 1C1 (SULT1C1) gene was differentially expressed between them. To investigate the differential expression mechanism of porcine SULT1C1 gene, we cloned its promoter region and predicted its activity. Six deletion expression vectors (P1, P2, P3, P4, P5, and P6) for the promoter of SULT1C1 gene were constructed. Vector P3 (-1084/+261) had the highest expression activity, whereas vector P4 (-642/+261) showed a reduced in promoter activity, which suggests that the core promoter region of SULT1C1 gene is located between -1084 bp and -642 bp. Two single nucleotide polymorphisms (SNPs), c. - 994 G > A (rs345070974) and c. - 946 G > A (rs337902009) were found in Mi and Large White gilts between -1100 and -661 bp, and the expression vectors with four haplotypes (GG, AA, GA, and AG) of two SNPs were constructed. The relative luciferase activity of vector with haplotype GG was the greatest among four vectors. These indicate that c. - 994 G > A and c. - 946 G > A are key mutations for promoter activity of SULT1C1 gene. Porcine SULT1C1 promoter with -994 G allele and -946 G allele significantly improved the gene expression level. It could be involved in different estrus expression between Large White and Mi gilts.
Assuntos
Sulfotransferases , Sus scrofa , Animais , Estrogênios , Feminino , Haplótipos/genética , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Sulfotransferases/genética , Sulfotransferases/metabolismo , Sus scrofa/genética , Suínos/genéticaRESUMO
Aggressive behavior has negative effects on animal welfare and growth performance in pigs. The dopamine receptor D2 (DRD2) has a critical neuromodulator role in the dopamine signal pathway within the brain to control behavior. A functional single-nucleotide polymorphism (SNP), rs1110730503, in the promoter region of the porcine DRD2 gene was identified, which affects aggressive behavior in pigs. A chromatin immunoprecipitation (ChIP) assay was used to identify the interactions between interferon regulatory factor 1 (IRF1) and IRF2 with the DRD2 gene. The overexpression or knockdown of these two transcription factors in porcine kidney-15 (PK15) and porcine neuronal cells (PNCs) indicate that the binding of IRF1 to DRD2 promotes the transcription of the DRD2 gene, but the binding of IRF2 to the DRD2 gene inhibits its transcription. Furthermore, IRF1 and IRF2 are functionally antagonistic to each other. The downregulation of DRD2 or upregulation of IRF2 increased the apoptosis rate of porcine neuroglial cells. Taken together, we found that transcriptional factors IRF1 and IRF2 have vital roles in regulating the transcription of the DRD2 gene, and rs1110730503 (-915A/T) is a functional SNP that influences IRF2 binding to the promoter of the DRD2 gene. These findings will provide further insight towards controlling aggressive behavior in pigs.
RESUMO
Porcine 17-hydroxysteroid dehydrogenase type 14 (HSD17B14) and FSH reporter (FSHR) genes play important roles in the metabolism of steroid hormones and the apoptosis of ovarian granulosa cells (GCs). Our bioinformatics analyses and the dual luciferase reporter assays indicated that porcine miR-20b and miR-31 target the 3'-UTR region of HSD17B14 gene, and miR-31 also targets the 3'-UTR region of FSHR gene. Overexpression of porcine HSD17B14 gene promoted the conversion from estradiol (E2) to estrone (E1) and increased the apoptosis of porcine GCs. Overexpression of miR-20b down-regulated the mRNA and protein expression level of HSD17B14 gene, decreased the concentration of progesterone (P4) and E1, increased E2, as well as reduced apoptosis of GCs. Moreover, overexpression of miR-31 also down-regulated the protein expression level of HSD17B14 gene, decreased the concentration of P4 and E1, and increased E2. However, miR-31 promoted apoptosis of GCs by targeting to the 3'-UTR of porcine FSHR gene. Taken together, we found that both porcine miR-20b and miR-31 target HSD17B14 gene, but miR-31 also targets FSHR gene to regulate the metabolism of steroid hormones and the apoptosis of porcine ovarian GCs. These findings expand the epigenetic regulatory mechanism of porcine miR-31 and miR-20b in ovarian GCs.
Assuntos
17-Hidroxiesteroide Desidrogenases/metabolismo , Apoptose , Células da Granulosa , MicroRNAs , Receptores do FSH/metabolismo , Animais , Estradiol , Estrona , Feminino , Células da Granulosa/citologia , MicroRNAs/genética , SuínosRESUMO
Copy number variation (CNV) has been considered to be an important source of genetic variation for important phenotypic traits of livestock. In this study, we performed whole-genome CNV detection on Suhuai (SH) (n = 23), Chinese Min Zhu (MZ) (n = 11), and Large White (LW) (n = 12) pigs based on next-generation sequencing data. The copy number variation regions (CNVRs) were annotated and analyzed, and 10,885, 10,836, and 10,917 CNVRs were detected in LW, MZ, and SH pigs, respectively. Some CNVRs have been randomly selected for verification of the variation type by real-time PCR. We found that SH and LW pigs are closely related, while MZ pigs are distantly related to the SH and LW pigs by CNVR-based genetic structure, PCA, VST, and QTL analyses. A total of 14 known genes annotated in CNVRs were unique for LW pigs. Among them, the cyclin T2 (CCNT2) is involved in cell proliferation and the cell cycle. The FA Complementation Group M (FANCM) is involved in defective DNA repair and reproductive cell development. Ten known genes annotated in 47 CNVRs were unique for MZ pigs. The genes included glycerol-3-phosphate acyltransferase 3 (GPAT3) is involved in fat synthesis and is essential to forming the glycerol triphosphate. Glutathione S-transferase mu 4 (GSTM4) gene plays an important role in detoxification. Eleven known genes annotated in 23 CNVRs were unique for SH pigs. Neuroligin 4 X-linked (NLGN4X) and Neuroligin 4 Y-linked (NLGN4Y) are involved with nerve disorders and nerve signal transmission. IgLON family member 5 (IGLON5) is related to autoimmunity and neural activities. The unique characteristics of LW, MZ, and SH pigs are related to these genes with CNV polymorphisms. These findings provide important information for the identification of candidate genes in the molecular breeding of pigs.
RESUMO
Visible and long-lasting estrus expression of gilts and sows effectively sends a mating signal. To reveal the roles of Long Intergenic Non-coding RNAs (lincRNAs) in estrus expression, RNA-seq was used to investigate the lincRNAs expression of follicular tissues from Large White gilts at diestrus (LD) and estrus (LE), and Chinese Mi gilts at diestrus (MD) and estrus (ME). Seventy-three differentially expressed lincRNAs (DELs) were found in all comparisons (LE vs. ME, LD vs. LE, and MD vs. ME comparisons). Eleven lincRNAs were differentially expressed in both LD vs. LE and MD vs. ME comparisons. Fifteen DELs were mapped onto the pig corpus luteum number Quantitative Trait Loci (QTL) fragments. A protein-protein interaction (PPI) network that involved estrus expression using 20 DEGs was then constructed. Interestingly, three predicted target DEGs (PTGs) (CYP19A1 of MSTRG.10910, CDK1 of MSTRG.10910 and MSTRG.23984, SCARB1 of MSTRG.1559) were observed in the PPI network. A competitive endogenous RNA (ceRNA) network including three lincRNAs, five miRNAs, and five genes was constructed. Our study provides new insight into the lincRNAs associated with estrus expression and follicular development in gilts.
RESUMO
Variation in genes of the serotonergic system influences aggressive behavior by affecting serotonin levels in the central and cortical nervous system. SLC6A4 (serotonin transporter) is a master regulator of 5-HT signaling and involved in the regulation of aggressive behavior in humans and rodents. To identify potential functional single nucleotide polymorphisms (SNPs) for the porcine SLC6A4 gene associated with aggressive behavior, a total of 500 pigs (268 barrows and 232 gilts) were selected and mixed in 51 pens. Their behavior was recorded and observed for 72 h after mixing. Based on a composite aggressive score (CAS), the most aggressive and the least aggressive pigs within each pen were selected separately (a total of 204 pigs). Ear tissue was sampled to extract genomic DNA. Eight SNPs in the 5'-flanking region, coding region, and 3'-untranslated region (3'-UTR) of SLC6A4 were genotyped, of which 6 SNPs had significant differences (Pâ <â 0.05) in allele frequency between the most aggressive and least aggressive pigs. Luciferase activity was greater in plasmids of genotype GG than plasmids of genotype CC of rs345058216 (Pâ <â 0.01). Computational analysis nominated MAZ as putative transcription factor (TF) with higher probability to bind the SLC6A4 promoter at the SNP (rs345058216) site. Also, we demonstrated that MAZ overexpression modulates SLC6A4 promoter activity in allele-specific manner with an in vitro assay. In addition, we demonstrated that SLC6A4 was a direct target of miR-671-5p. The dual luciferase reporter gene assay and cell transfection were performed to examine the role of miR-671-5p in regulating SLC6A4 expression. The luciferase assays revealed that the SNP rs332335871 affects regulation of miR-671-5p in SLC6A4 expression. After overexpression of miR-671-5p in porcine primary neural cells, the SLC6A4 mRNA levels can be significantly reduced. In conclusion, we here found that miR-671-5p and MAZ mediated porcine SLC6A4 expression level, which provides the possible molecular mechanism of aggressive behavior.
The current study identified the functional single nucleotide polymorphisms for the porcine SLC6A4 (serotonin transporter) gene associated with aggressive behavior of pigs after mixing. We investigated the underlying molecular regulation mechanism of those functional variations. SNP rs345058216 (c.-1694Câ >â G) affects binding of the transcription factor MAZ (MYC associated zinc finger protein) to the core promoter region of the SLC6A4 gene, whereas SNP rs332335871 (c.*1586Gâ >â A) affects the binding of miR-671-5p to the 3ʹ-UTR of SLC6A4 gene. These results suggest that rs345058216 and rs332335871 could be used as candidate molecular markers for aggressive behavior of pigs after mixing.