RESUMO
Adipose tissue plays critical roles in an individual's aging process. In this research, we use single-nucleus RNA sequencing to create highly detailed transcriptional maps of subcutaneous adipose tissue and visceral adipose tissue in young and aged mice. We comprehensively identify the various cell types within the white adipose tissue of mice, our study has elucidated seven distinct cell types within this tissue. Further analyses focus on adipocytes, fibro-adipogenic progenitors, and immune cells, revealing age-related declines in the synthetic metabolic activity of adipocytes, diminished immune regulation, and reduced maturation or proliferation of fibroblasts in undifferentiated adipocytes. We confirm the presence of distinct subpopulations of adipocytes, highlighting decreases in adipogenesis subgroups due to aging. Additionally, we uncover a reduction in immune cell subpopulations, driven by age-associated immune system dysregulation. Furthermore, pseudo-time analyses indicate that Adipocyte1 represents the 'nascent' phase of adipocyte development, while Adipocyte2 represents the 'mature' phase. We use cell-cell interaction to explore the age-dependent complexities of the interactions between FAPs and adipocytes, and observed increased expression of the inflammation-related Retn-Tlr4 interaction in older mice, while the anti-inflammatory Angpt1-Tek interaction was only detected in young mice. These transcriptional profiles serve as a valuable resource for understanding the functional genomics underlying metabolic disorders associated with aging in human adipose tissue.
Assuntos
Adipócitos , Envelhecimento , Perfilação da Expressão Gênica , Animais , Envelhecimento/genética , Camundongos , Adipócitos/metabolismo , Transcriptoma , Adipogenia/genética , Tecido Adiposo/metabolismo , Gordura Intra-Abdominal/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Tecido Adiposo Branco/metabolismo , Análise de Célula ÚnicaRESUMO
In diploid mammals, allele-specific three-dimensional (3D) genome architecture may lead to imbalanced gene expression. Through ultradeep in situ Hi-C sequencing of three representative somatic tissues (liver, skeletal muscle, and brain) from hybrid pigs generated by reciprocal crosses of phenotypically and physiologically divergent Berkshire and Tibetan pigs, we uncover extensive chromatin reorganization between homologous chromosomes across multiple scales. Haplotype-based interrogation of multi-omic data revealed the tissue dependence of 3D chromatin conformation, suggesting that parent-of-origin-specific conformation may drive gene imprinting. We quantify the effects of genetic variations and histone modifications on allelic differences of long-range promoter-enhancer contacts, which likely contribute to the phenotypic differences between the parental pig breeds. We also observe the fine structure of somatically paired homologous chromosomes in the pig genome, which has a functional implication genome-wide. This work illustrates how allele-specific chromatin architecture facilitates concomitant shifts in allele-biased gene expression, as well as the possible consequential phenotypic changes in mammals.
Assuntos
Cromatina , Cromossomos , Animais , Suínos/genética , Cromatina/genética , Haplótipos , Cromossomos/genética , Genoma , Mamíferos/genéticaRESUMO
BACKGROUND: Femoral neck fractures represent a significant public health concern, particularly in the elderly population. A thorough understanding and assessment of these fractures are deemed essential for optimal treatment and management. Displacement characteristics of Garden III femoral neck fractures were explored in this study, and the reliability, validity, and clinical utility of the anteroposterior Garden Index in evaluating displacement severity were investigated. METHODS: Patients diagnosed with Garden III femoral neck fractures were included in this study. The anteroposterior Garden Index was computed from X-ray images by three experienced orthopedic doctors. Additionally, the contact area of the fracture endpoint and displacement of the femoral neck were evaluated using 128-slice 3D CT scans. Inter-observer and retest reliability of the Garden Index measurements were assessed, along with its correlation with CT measurements. RESULTS: In this study, a total of 110 patients with Garden III femoral neck fractures were analyzed, showcasing an almost equal gender distribution and an age range spanning from 20 to 88 years. An average Garden Index of 135° (± 16°) was observed. The intra-observer repeatability of the Garden Index was found to exceed 90%. A significant positive correlation was identified between the Garden Index and the contact surface area of the fracture endpoint (r = 0.82, P < 0.001), while a significant negative correlation was noted with the upward displacement of the femoral neck (r = - 0.79, P < 0.001). CONCLUSIONS: The anteroposterior Garden Index has been demonstrated to have promising potential as a reliable and valid tool for assessing the displacement severity of Garden III femoral neck fractures. Nonetheless, further research is needed to elucidate its relationship with other fracture characteristics and to enhance its criterion and construct validity.
Assuntos
Fraturas do Colo Femoral , Humanos , Idoso , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Reprodutibilidade dos Testes , Fraturas do Colo Femoral/diagnóstico por imagem , Fraturas do Colo Femoral/cirurgia , Colo do Fêmur , Tomografia Computadorizada por Raios X , Fixação Interna de Fraturas/métodosRESUMO
Using an adult female miniature pig model with diet-induced weight gain/weight loss, we investigated the regulatory mechanisms of three-dimensional (3D) genome architecture in adipose tissues (ATs) associated with obesity. We generated 249 high-resolution in situ Hi-C chromatin contact maps of subcutaneous AT and three visceral ATs, analyzing transcriptomic and chromatin architectural changes under different nutritional treatments. We find that chromatin architecture remodeling underpins transcriptomic divergence in ATs, potentially linked to metabolic risks in obesity development. Analysis of chromatin architecture among subcutaneous ATs of different mammals suggests the presence of transcriptional regulatory divergence that could explain phenotypic, physiological, and functional differences in ATs. Regulatory element conservation analysis in pigs and humans reveals similarities in the regulatory circuitry of genes responsible for the obesity phenotype and identified non-conserved elements in species-specific gene sets that underpin AT specialization. This work provides a data-rich tool for discovering obesity-related regulatory elements in humans and pigs.
Assuntos
Cromatina , Aumento de Peso , Adulto , Humanos , Feminino , Suínos , Animais , Obesidade , Tecido Adiposo , Montagem e Desmontagem da Cromatina , Redução de Peso , MamíferosRESUMO
To explore the latent classes of stigma in patients with rheumatoid arthritis, we analyzed the characteristics of the different categories. Adopting a convenient sampling method, socio-demographic and disease-related information from the outpatient clinics and wards of 3 tertiary care hospitals in China was collected. The Chinese version of the Internalized Stigma of Mental Illness scale-Rheumatoid Arthritis was used in this survey. Rheumatoid arthritis stigma was divided into 3 potential categories: Low Stigma-Strong Resistance (83, 41.5%), Medium Stigma-Strong Alienation (78, 39.0%), and High Stigma-Weak Resistance (39, 19.5%). Unordered multinomial logistic regression analysis showed that pain (OR = 1.540, P = .005; OR = 1.797, P < .001), elementary school education and below (OR = 4.051, P = .037), and duration of morning stiffness (OR = 0.267, P = .032) were risk factors for stigma, whereas family history was a protective factor against stigma (OR = 0.321, P = .046). Patients with longer morning stiffness, more severe pain, and less education have a greater risk of heavier stigma. Strong alienation is an early warning of heavy stigma. Resistance to stigma and family support can help patients overcome their psychological obstacles. More attention should be paid to constructing family centered support systems to help resist stigma.
Assuntos
Artrite Reumatoide , Estigma Social , Humanos , Análise de Classes Latentes , Artrite Reumatoide/psicologia , Fatores de Risco , DorRESUMO
Diabetic wound healing has become a serious healthcare challenge. The high-glucose environment leads to persistent bacterial infection and mitochondrial dysfunction, resulting in chronic inflammation, abnormal vascular function, and tissue necrosis. To solve these issues, we developed a double-network hydrogel, constructed with pluronic F127 diacrylate (F127DA) and hyaluronic acid methacrylate (HAMA), and enhanced by SS31-loaded mesoporous polydopamine nanoparticles (MPDA NPs). As components, SS31, a mitochondria-targeted peptide, maintains mitochondrial function, reduces mitochondrial reactive oxygen species (ROS) and thus regulates macrophage polarization, as well as promoting cell proliferation and migration, while MPDA NPs not only scavenge ROS and exert an anti-bacterial effect by photothermal treatment under near-infrared light irradiation, but also control release of SS31 in response to ROS. This F127DA/HAMA-MPDA@SS31 (FH-M@S) hydrogel has characteristics of adhesion, superior biocompatibility and mechanical properties which can adapt to irregular wounds at different body sites and provide sustained release of MPDA@SS31 (M@S) NPs. In addition, in a diabetic rat full thickness skin defect model, the FH-M@S hydrogel promoted macrophage M2 polarization, collagen deposition, neovascularization and wound healing. Therefore, the FH-M@S hydrogel exhibits promising therapeutic potential for skin regeneration.
RESUMO
BACKGROUND: Spinal cord injury (SCI) is a traumatic central nervous system disorder that leads to irreversible neurological dysfunction. Emerging evidence has shown that differentially expressed circular RNAs (circRNAs) after SCI is closely associated with the pathophysiological process. Herein, the potential function of circRNA spermine oxidase (circSmox) in functional recovery after SCI was investigated. METHODS: Differentiated PC12 cells stimulated with lipopolysaccharide (LPS) were employed as an in vitro model for neurotoxicity research. Levels of genes and proteins were detected by quantitative real-time PCR and Western blot analysis. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry. Western blot analysis was used to detect the protein level of apoptosis-related markers. The levels of interleukin (IL)-1ß, IL-6, IL-8, and tumor necrosis factor (TNF)-α. Dual-luciferase reporter, RIP, and pull-down assays were used to confirm the target relationship between miR-340-5p and circSmox or Smurf1 (SMAD Specific E3 Ubiquitin Protein Ligase 1). RESULTS: LPS elevated the levels of circSmox and Smurf1, but decreased the levels of miR-340-5p in PC12 cells in a dose-dependent manner. Functionally, circSmox silencing alleviated LPS-induced apoptosis and inflammation in PC12 cells in vitro. Mechanistically, circSmox directly sponged miR-340-5p, which targeted Smurf1. Rescue experiments showed that miR-340-5p inhibition attenuated the neuroprotective effect of circSmox siRNA in PC12 cells. Moreover, miR-340-5p suppressed LPS-triggered neurotoxicity in PC12 cells, which was reversed by Smurf1 overexpression. CONCLUSION: CircSmox enhances LPS-induced apoptosis and inflammation via miR-340-5p/Smurf1 axis, providing an exciting view of the potential involvement of circSmox in SCI pathogenesis.
Assuntos
MicroRNAs , Traumatismos da Medula Espinal , Animais , Ratos , Apoptose/genética , Inflamação/genética , Lipopolissacarídeos/toxicidade , MicroRNAs/genética , Células PC12 , Traumatismos da Medula Espinal/genética , Ubiquitina-Proteína Ligases/genética , Poliamina OxidaseRESUMO
The objective of this article is to assess self-reported sleep disturbance and identify psychological, clinical, and sociodemographic factors that might influence sleep disturbance in patients with rheumatoid arthritis (RA). The study included 141 patients with confirmed RA (84.4% women, mean age 56.87 years). The Pittsburgh Sleep Quality Index, the Chinese version of rheumatoid arthritis self-efficacy scale, the Chinese version of Anxiety Depression Distress Inventory-27, the Chinese version of Stigma Scale for Chronic Illness, Visual Analogue Scale-Pain, disease activity index were used. Sleep disturbance was positively correlated with age, pain, disease activity, depression and anxiety, and stigma, while self-efficacy was correlated negatively with sleep disturbance. Multiple linear regression analysis revealed that depression, anxiety, self-efficacy, and stigma explained 77.4% of sleep quality variance. The data has demonstrated a suggestive relationship between low sleep quality and anxiety, depression, self-efficacy, and stigma. Patients reporting poor sleep, fatigue, and pain might have particular psychological intervention needs focusing on distress or anxiety symptoms, low self-efficacy, and high stigma.
Assuntos
Artrite Reumatoide , Transtornos do Sono-Vigília , Humanos , Feminino , Pessoa de Meia-Idade , Masculino , Autorrelato , Depressão/psicologia , Autoeficácia , Qualidade de Vida/psicologia , Artrite Reumatoide/psicologia , Ansiedade/epidemiologia , Ansiedade/psicologia , Transtornos do Sono-Vigília/epidemiologia , Transtornos do Sono-Vigília/psicologia , Dor/epidemiologia , Dor/psicologia , Sono , Fadiga/psicologiaRESUMO
Objective: To investigate the short-term effectiveness and advantages of the orthopedic robot-assisted femoral neck system (FNS) fixation in the treatment of fresh femoral neck fractures compared with the traditional manual operation. Methods: A clinical data of 74 patients with fresh femoral neck fractures, who had undergone internal fixation with FNS between April 2020 and September 2021, was retrospectively analyzed. Among them, there were 31 cases of TiRobot-assisted operation (trial group) and 43 cases of traditional manual operation (control group). There was no significant difference between groups ( P>0.05) in terms of gender, age, cause of injury, time from injury to operation, fracture side and type. The fracture fixation time (intraoperative fracture reduction to the end of suture), invasive fixation time (incision of internal fixation to the end of suture), the number of placing key-guide needle, incision length, intraoperative blood loss, fracture healing, and Harris score of hip function were recorded and compared. Results: All operations were performed with no neurovascular injury or incision complications. The invasive fixation time, intraoperative blood loss, the number of placing key-guide needle, and the incision length in the trial group were superior to the control group ( P<0.05), and there was no significant difference in fracture fixation time between groups ( P>0.05). All patients were followed up 4-16 months (mean, 7 months). The fracture did not heal in 1 patient of trial group, and the other fractures healed in 2 groups; the fracture healing time was (17.6±1.9) weeks in trial group and (18.2±1.9) weeks in control group, and there was no significant difference between groups ( t=0.957, P=0.345). At last follow-up, the Harris score of the trial group was 82.4±5.8, which was higher than that of the control group (79.0±7.7), but the difference was not significant ( t=-1.483, P=0.147). Conclusion: Orthopedic robot-assisted FNS fixation in the treatment of fresh femoral neck fractures has the similar short-term effectiveness as the traditional method, but the former has advantages in terms of operation time, intraoperative blood loss, and the number of placing key-guide needle, making the operation more minimally invasive and quicker, and more suitable for older patients.
Assuntos
Fraturas do Colo Femoral , Robótica , Perda Sanguínea Cirúrgica , Fraturas do Colo Femoral/cirurgia , Colo do Fêmur , Fixação Interna de Fraturas/métodos , Humanos , Estudos Retrospectivos , Resultado do TratamentoRESUMO
BACKGROUND: Gliomas are the most frequent and aggressive cancers in the central nervous system, and spinal cord glioma (SCG) is a rare class of the gliomas. Empty spiracles homobox genes (EMXs) have shown potential tumor suppressing roles in glioma, but the biological function of EMX1 in SCG is unclear. METHODS: The EMX1 expression in clinical tissues of patients with SCG was examined. SCG cells were extracted from the tissues, and altered expression of EMX1 was then introduced to examine the role of EMX1 in cell growth and invasiveness in vitro. Xenograft tumors were induced in nude mice for in vivo validation. The targets of EXM1 were predicted via bioinformatic analysis and validated by luciferase and ChIP-qPCR assays. Rescue experiments were conducted to validate the involvements of the downstream molecules. RESULTS: EMX1 was poorly expressed in glioma, which was linked to decreased survival rate of patients according to the bioinformatics prediction. In clinical tissues, EMX1 was poorly expressed in SCG, especially in the high-grade tissues. EMX1 upregulation significantly suppressed growth and metastasis of SCG cells in vitro and in vivo. EMX1 bound to the promoter of WASP family member 2 (WASF2) to suppress its transcription. Restoration of WASF2 blocked the tumor-suppressing effect of EMX1. EMX1 suppressed Wnt/ß-catenin signaling activity by inhibiting WASF2. Coronaridine, a Wnt/ß-catenin-specific antagonist, blocked SCG cell growth and metastasis induced by WASF2. CONCLUSION: This study elucidates that EMX1 functions as a tumor inhibitor in SCG by suppressing WASF2-dependent activation of the Wnt/ß-catenin axis.
Assuntos
Glioma , Proteínas de Homeodomínio/metabolismo , Fatores de Transcrição/metabolismo , beta Catenina , Animais , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Glioma/tratamento farmacológico , Glioma/genética , Humanos , Camundongos , Camundongos Nus , Medula Espinal , Família de Proteínas da Síndrome de Wiskott-Aldrich/genética , Família de Proteínas da Síndrome de Wiskott-Aldrich/metabolismo , beta Catenina/genética , beta Catenina/metabolismoRESUMO
PURPOSE: To detect the expression level of hsa_circ_0005721 in osteosarcoma specimen and plasma of osteosarcoma patients, and to analyze the clinical significance of hsa_circ_0005721 as a diagnostic marker for osteosarcoma. METHODS: Expression levels of hsa_circ_0005721 in osteosarcoma specimen and osteosarcoma cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between hsa_circ_0005721 expression difference and overall survival in osteosarcoma was analyzed by Kaplan-Meier method. Expression level of hsa_circ_0005721 was stably downregulated in U-2OS and HOS cells by shRNA transfection. Proliferative potential in osteosarcoma cells regulated by hsa_circ_0005721 was assessed by colony formation and 5-Ethynyl-2'- deoxyuridine (EdU) assay. Differentially expressed hsa_circ_0005721 in the plasma of healthy controls, benign bone tumor patients and osteosarcoma patients was determined by qRT-PCR. The diagnostic capacity of hsa_circ_0005721 in osteosarcoma was examined by receiver operating characteristic (ROC) curves. RESULTS: hsa_circ_0005721 was upregulated in osteosarcoma specimen and osteosarcoma cell lines. High level of hsa_circ_0005721 predicted poor prognosis in osteosarcoma patients. In vitro experiments showed that knockdown of hsa_circ_0005721 suppressed proliferative ability in osteosarcoma cells. Compared with that in healthy controls and benign bone tumor patients, plasma level of hsa_circ_0005721 was higher in osteosarcoma patients. ROC curves demonstrated the diagnostic potential of hsa_circ_0005721 in osteosarcoma. CONCLUSIONS: hsa_circ_0005721 is upregulated in osteosarcoma samples, which acts as an oncogene responsible for aggravating the progression. hsa_circ_0005721 can be a promising diagnostic marker for osteosarcoma.
Assuntos
Biomarcadores Tumorais/análise , Neoplasias Ósseas/sangue , Neoplasias Ósseas/química , Osteossarcoma/sangue , Osteossarcoma/química , RNA Circular/análise , Biomarcadores Tumorais/genética , Neoplasias Ósseas/diagnóstico , Humanos , Osteossarcoma/diagnóstico , RNA Circular/genética , Células Tumorais CultivadasRESUMO
The existing research on dairy cow mammary gland genes is extensive, but there have been few reports about dynamic changes in dairy cow mammary gland genes as milk yield decrease. For the first time, transcriptome analysis based on short time-series expression miner (STEM) and histological observations were performed using the Holstein dairy cow mammary gland to explore gene expression patterns in this process of decrease (at peak, mid-, and late lactation). Histological observations suggested that the number of mammary acinous cells at peak/mid-lactation was significantly higher than that at mid-/late lactation, and the lipid droplets area secreted by dairy cows was almost unaltered across the three stages of lactation (p > 0.05). Totals of 882 and 1439 genes were differentially expressed at mid- and late lactation, respectively, compared to peak lactation. Function analysis showed that differentially expressed genes (DEGs) were mainly related to apoptosis and energy metabolism (fold change ≥ 2 or fold change ≤ 0.5, p-value ≤ 0.05). Transcriptome analysis based on STEM identified 16 profiles of differential gene expression patterns, including 5 significant profiles (false discovery rate, FDR ≤ 0.05). Function analysis revealed DEGs involved in milk fat synthesis were downregulated in Profile 0 and DEGs in Profile 12 associated with protein synthesis. These findings provide a foundation for future studies on the molecular mechanisms underlying mammary gland development in dairy cows.
Assuntos
Bovinos/genética , Lactação , Glândulas Mamárias Animais/metabolismo , Transcriptoma , Animais , Bovinos/fisiologia , Metabolismo Energético , FemininoRESUMO
Staphylococcus aureus- induced mastitis is one of the most intractable problems for the dairy industry, which causes loss of milk yield and early slaughter of cows worldwide. Few studies have used a comprehensive approach based on the integrative analysis of miRNA and mRNA expression profiles to explore molecular mechanism in bovine mastitis caused by S. aureus. In this study, S. aureus (A1, B1 and C1) and sterile phosphate buffered saline (PBS) (A2, B2 and C2) were introduced to different udder quarters of three individual cows, and transcriptome sequencing and microarrays were utilized to detected miRNA and gene expression in mammary glands from the challenged and control groups. A total of 77 differentially expressed microRNAs (DE miRNAs) and 1625 differentially expressed genes (DEGs) were identified. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that multiple DEGs were enriched in significant terms and pathways associated with immunity and inflammation. Integrative analysis between DE miRNAs and DEGs proved that miR-664b, miR-23b-3p, miR-331-5p, miR-19b and miR-2431-3p were potential factors regulating the expression levels of CD14 Molecule (CD14), G protein subunit gamma 2 (GNG2), interleukin 17A (IL17A), collagen type IV alpha 1 chain (COL4A1), microtubule associated protein RP/EB family member 2 (MAPRE2), member of RAS oncogene family (RAP1B), LDOC1 regulator of NFKB signaling (LDOC1), low-density lipoprotein receptor (LDLR) and S100 calcium binding protein A9 (S100A9) in bovine mastitis caused by S. aureus. These findings could enhance the understanding of the underlying immune response in bovine mammary glands against S. aureus infection and provide a useful foundation for future application of the miRNA-mRNA-based genetic regulatory network in the breeding cows resistant to S. aureus.
RESUMO
Cardiovascular diseases (CVDs) are seriously threatening to human life and health. Polyunsaturated fatty acids (PUFAs) are known for their role in preventing CVDs. It is beneficial to population health to promote the content of PUFAs in bovine milk. In recent years, limited research based on molecular mechanisms has focused on this field. The biological roles of numerous microRNAs (miRNAs) remain unknown. In this study, a promising and negatively correlated pair of the miRNA (miRNA-193a-5p) and a fatty acid desaturase 1 (FADS1) gene are identified and screened to explore whether they are potential factors of PUFAs' synthesis in bovine milk. The targeted relationship between miRNA-193a-5p and FADS1 in bovine mammary epithelial cells (BMECs) is demonstrated by dual luciferase reporter assays. qRT-PCR and western blot assays indicate that both the expression of mRNA and the protein FADS1 show a negative correlation with miRNA-193a-5p expression in BMECs. Also, miR-193a-5p expression is positively correlated with the expression of genes associated with milk fatty acid metabolism, including ELOVL fatty acid elongase 6 (ELOVL6) and diacylglycerol O-acyltransferase 2 (DGAT2). The expression of fatty acid desaturase 2 (FADS2) is negatively correlated with miR-193a-5p expression in BMECs. The contents of triglycerides (TAG), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) have a significant positive correlation with the expression of FADS1 and a significant negative correlation with the expression of miR-193a-5p in BMECs. For the first time, this study confirms that miRNA-193a-5p regulates PUFAs metabolism in BMECs by targeting FADS1, indicating that miRNA-193a-5p and FADS1 are underlying factors that improve PUFAs content in bovine milk.
Assuntos
Células Epiteliais/citologia , Ácidos Graxos Dessaturases/metabolismo , Regulação da Expressão Gênica , Glândulas Mamárias Animais/metabolismo , MicroRNAs/genética , Animais , Bovinos , Dessaturase de Ácido Graxo Delta-5 , Diacilglicerol O-Aciltransferase/biossíntese , Elongases de Ácidos Graxos/biossíntese , Ácidos Graxos Insaturados/metabolismo , Feminino , Perfilação da Expressão Gênica , Metabolismo dos Lipídeos/genética , Leite , Análise de Componente Principal , Triglicerídeos/metabolismoRESUMO
Weighted gene coexpression network analysis (WGCNA) is a novel approach that can quickly analyze the relationships between genes and traits. In this study, the milk yield, lactose, fat, and protein of Holstein dairy cows were detected in a lactation cycle. Meanwhile, a total of 18 gene expression profiles were detected using mammary glands from six lactation stages (day 7 to calving, -7 d; day 30 post-calving, 30 d; day 90 post-calving, 90 d; day 180 post-calving, 180 d; day 270 post-calving, 270 d; day 315 post-calving, 315 d). On the basis of the 18 profiles, WGCNA identified for the first time 10 significant modules that may be related to lactation stage, milk yield, and the main milk composition content. Genes in the 10 significant modules were examined with gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The results revealed that the galactose metabolism pathway was a potential candidate for milk yield and milk lactose synthesis. In -7 d, ion transportation was more frequent and cell proliferation related terms became active. In late lactation, the suppressor of cytokine signaling 3 (SOCS3) might play a role in apoptosis. The sphingolipid signaling pathway was a potential candidate for milk fat synthesis. Dairy cows at 315 d were in a period of cell proliferation. Another notable phenomenon was that nonlactating dairy cows had a more regular circadian rhythm after a cycle of lactation. The results provide an important theoretical basis for the further molecular breeding of dairy cows.
RESUMO
The concentration of bovine milk fat changes regularly with lactation stages. In particular, milk fat percentage is higher in late lactation than mid lactation. Furthermore, milk fat composition is highly subject to a few genes. Thus, transcriptome sequencing was performed to explore the expression patterns of differentially-expressed genes (DEGs) in the parenchymal mammary gland of Holstein dairy cows between mid and late lactation. The 725 DEGs were screened (fold change > 2 and p-value < 0.05), and the peroxisome proliferator-activated receptor (PPAR) signaling pathway associated with lipid synthesis had a significant variation between the two periods (p-value < 0.05). The activation of the PPAR signal pathway may a key factor in the increasing of milk fat content in late lactation compared to mid lactation. Acyl-CoA synthetase long-chain family member 4 (ACSL4), a member of the PPAR signaling pathway, was upregulated in late lactation compared to mid lactation (p < 0.05). ACSL4 catalyzes the activation of long-chain fatty acids for cellular lipid synthesis. However, it remains uncertain that the molecular mechanism of milk fat synthesis is regulated by ACSL4 in dairy cows. Subsequently, the function verification of ACSL4 was performed in bovine mammary epithelial cells (BMECs). The upregulated expression of ACSL4 was accompanied by the increase of the concentration of intracellular triglycerides, whereas knockdown of ACSL4 decreased the concentration of intracellular triglycerides, which demonstrated that ACSL4 plays an important role in modulating milk fat synthesis. In conclusion, the results displayed that ACSL4 expression regulates triglyceride metabolism in ruminant mammary cells.
Assuntos
Coenzima A Ligases/genética , Lactação/fisiologia , Glândulas Mamárias Animais/metabolismo , Leite/metabolismo , Animais , Bovinos , Coenzima A Ligases/metabolismo , Células Epiteliais/metabolismo , Feminino , Expressão Gênica , Lactação/genética , Metabolismo dos Lipídeos/genética , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/fisiologia , Triglicerídeos/genética , Triglicerídeos/metabolismoRESUMO
The beef aging process is essential for compliance with certain major requisites, such as sensory characteristics for cooking and meat processing. Meat quality analysis of Yunling cattle, a new hybrid beef cattle bred by Chinese researchers, during the aging process, represents a major research gap. To explore Yunling beef initially, indicators associated with meat quality during the aging process of Yunling, Simmental, and Wenshan cattle were measured. In addition, some important economic traits were detected in the three breeds, including growth performance and carcass characteristics. The results showed that the growth performance, carcass traits, pH, and water holding capacity of Yunling and Simmental cattle were basically the same and better, respectively, than those of Wenshan cattle. The proportions of individual fatty acids in Yunling beef were healthier than in the other two breeds. Aging time did not affect the fatty acid profiles of the beef (p > 0.05). The contents of certain fatty acids in the three beef types displayed some differences in terms of days of aging (p < 0.05). The tenderness and meat color were better in the Yunling beef as the aging time increased, indicating that Yunling beef aged for 7 days was more suitable for cooking, exhibiting better sensory characteristics. Thus, a 7-day short-term aging process is very effective in improving the quality of Yunling beef. Our study attempted to fill a gap in the Yunling beef quality analysis during aging, providing further evidence for Yunling beef improvement.
RESUMO
The hypothalamic-pituitary-thyroid (HPT) axis hormones regulate the growth and development of ruminants, and the pituitary gland plays a decisive role in this process. In order to identify pivotal genes in the pituitary gland that could affect the growth of cattle by regulating the secretion of hormones, we detected the content of six HPT hormones related to growth in the plasma of two cattle breeds (Yunling and Leiqiong cattle, both also known as the zebu cattle) with great differences in growth and compared the transcriptome data of their pituitary glands. Our study found that the contents of GH, IGF, TSH, thyroxine, triiodothyronine, and insulin were significantly different between the two breeds, which was the main cause of the difference in growth; 175 genes were identified as differentially expressed genes (DEGs). Functional association analyses revealed that DEGs were mainly involved in the process of transcription and signal transduction. Combining the enrichment analysis and protein interaction analysis, eight DEGs were predicted to control the growth of cattle by affecting the expression of growth-related hormones in the pituitary gland. In summary, our results suggested that SLC38A1, SLC38A3, DGKH, GNB4, GNAQ, ESR1, NPY, and GAL are candidates in the pituitary gland for regulating the growth of Yunling and Leiqiong cattle by regulating the secretion of growth-related hormones. This study may help researchers further understand the growth mechanisms and improve the artificial selection of zebu cattle.
RESUMO
Somatic cell count (SCC) in milk is widely used in the dairy industry, as an indicator of the health of mammary gland. While the SCC of dairy cattle was higher in late lactation than in peak lactation, its association with gene expressions of mammary gland were largely unknown. In this study, a transcriptomic sequencing approach and bioinformatics analysis were used to investigate the differential expressed genes (DEGs) associated with inflammation and immunity between peak and late periods of lactation in Chinese Holstein. A total of 446 DEGs (padj < 0.05 and fold change >2) were identified, 50 of which belonged to seven pathways and five terms related to inflammation and immunity. Our data suggested that the activation of nuclear transcription factor-κB (NF-κB) pathway and Toll-like receptor signaling pathway caused inflammatory response, and the activation of chemokine signaling pathway and cytokine-cytokine receptor interaction signaling pathway caused a protective immune response to ensure dairy cows health during late lactation. Our findings deepen the understanding of the molecular mechanism and physiological functions of mammary inflammation in Chinese Holstein during late lactation.
RESUMO
Cell polarity is the asymmetric compartmentalization of cellular components. An opposing gradient of partitioning-defective protein kinases, atypical protein kinase C (aPKC) and PAR-1, at the cell cortex guides diverse asymmetries in the structure of metazoan cells, but the mechanism underlying their spatial patterning remains poorly understood. Here, we show in Caenorhabditis elegans zygotes that the cortical PAR-1 gradient is patterned as a consequence of dual mechanisms: stabilization of cortical dynamics and protection from aPKC-mediated cortical exclusion. Dual control of cortical PAR-1 depends on a physical interaction with the PRBH-domain protein PAR-2. Using a reconstitution approach in heterologous cells, we demonstrate that PAR-1, PAR-2, and polarized Cdc42-PAR-6-aPKC comprise the minimal network sufficient for the establishment of an opposing cortical gradient. Our findings delineate the mechanism governing cortical polarity, in which a circuit consisting of aPKC and the PRBH-domain protein ensures the local recruitment of PAR-1 to a well-defined cortical compartment.