Background： The p21-activated kinase 1 （PAK1）is essential of microtubule assembly during oocyte meiotic maturation porcine oocytes. for mitosis and plays an important role in the regulatio in mice; however, little ...Background： The p21-activated kinase 1 （PAK1）is essential of microtubule assembly during oocyte meiotic maturation porcine oocytes. for mitosis and plays an important role in the regulatio in mice; however, little is known about its role in Result： Total p21-activated kinase 1 （PAK1） and phosphorylated PAK1 at Thr423 （PAK1^Thr423） were consistently expressed in porcine oocytes from the germinal vesicle （GV） to the second metaphase （MII） stages, but phosphorylation of histone H3 at Serr10 （H3^ser10） was only expressed after the GV stage. Immunofiuorescence analysis revealed that PAK1Thr423 and H3^ser10 colocalized on chromosomes after the GV stage. Blocking of endogenous PAK1^Thr423 by injecting a specific antibody decreased the phosphorylation level of H3^ser10; however, it had no impact on chromatin condensation, meiotic progression, cleavage rate of blastomeres or the rate of blastocyst formation. Conclusion： Phosphorylation of PAK1^Thr423 is a spontaneous activation process and the activated PAK1^Thr423 can promote the phosphorylation of H3^ser10; however, this pathway is not required for meiotic maturation of porcine oocytes or early embryonic development.展开更多
Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have bee...Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have been shown to be important regulators of GCs function. miR-34c involved in the regulation of various biological processes and was identified to be a pro-apoptotic and anti-proliferative factor in many cell types. However, the roles of miR-34c in GCs function remain unknown. In this study, we used Annexin V-FITC and Ed U assays to demonstrate that miR-34c exerted pro-apoptotic and anti-proliferative effects in porcine GCs. Dual-luciferase reporter assays, quantitative real-time PCR（q RT-PCR） and Western blotting identified Forkhead box O3a（Fox O3a） as a direct target gene of miR-34c. The overexpression of FoxO3a rescued the phenotypic change caused by miR-34c in porcine GCs. In conclusion, miR-34c regulate the function of porcine GCs by targeting FoxO3a.展开更多
The development history, etiology, epidemiology, clinical symptoms, pathological changes and diagnosis technology of porcine circovirus 3 were summarized, and countermeasures for prevention and control of the disease ...The development history, etiology, epidemiology, clinical symptoms, pathological changes and diagnosis technology of porcine circovirus 3 were summarized, and countermeasures for prevention and control of the disease were put forward based on the prevalence status in China.展开更多
Porcine placenta extract (PPE) is widely accepted as an ingredient in complementary and alternative medicine and previous studies have reported its availability, however, its underlying action mechanism remains unclea...Porcine placenta extract (PPE) is widely accepted as an ingredient in complementary and alternative medicine and previous studies have reported its availability, however, its underlying action mechanism remains unclear. In this study, we investigated the underlying mechanism of PPE-induced ceramide synthase 3 upregulation. PPE enhanced the expression of ceramide synthase 3 at both the mRNA and protein levels in HaCaT cells. Moreover, PPE-induced ceramide synthase3 upregulation was suppressed by the Akt inhibitor, suggesting the involvement of Akt in the underlying mechanism. As the PI3K inhibitor did not affect PPE-induced ceramide synthase 3 upregulation, the factors upstream of Akt were estimated. Inhibition and small interfering RNA experiments demonstrated that the peroxisome proliferator-activated receptors δ (PPARδ) and integrin-linked kinase (ILK) are involved in the phosphorylation of Akt. Next, we explored the factors downstream of Akt and found that PPE induced phosphorylation of the STAT3 while PPE-induced upregulation of ceramide synthase 3 was significantly suppressed by the inhibitor of the mTOR, suggesting that the mTOR/STAT3 pathway is involved in the downstream of Akt. These results demonstrate that PPE upregulates CerS3 expression via the PPARδ/ILK/Akt/mTOR/STAT3 pathway.展开更多
Porcine circovirus type 3(PCV3),which was first detected in the United States of America in 2015,is a potential threat to the swine industry.However,the prevalence of PCV3 in Shanxi Province,China,is unclear.In this r...Porcine circovirus type 3(PCV3),which was first detected in the United States of America in 2015,is a potential threat to the swine industry.However,the prevalence of PCV3 in Shanxi Province,China,is unclear.In this research,the prevalence and genetic diversity of PCV3 were investigated in above area.Lung tissue samples(n=491)from 19 pig slaughterhouses across 11 cities throughout Shanxi Province were analyzed for PCV3 infection by PCR in 2019.The results showed that PCV3 positive rates in slaughterhouses and individuals were 100%(19/19)and 86.76%(426/491),respectively.PCV2 and PCV3 double-positive rates in slaughterhouses and individuals were 100%(19/19)and 59.27%(291/491),respectively.PCR positive samples were further sequenced and 8 PCV3 isolates were identified.The nucleotide homology of these isolates with other PCV3 isolates in NCBI database was 97.45-99.90%.A phylogenetic analysis,based on the complete genomic sequence and ORF2,divided these PCV3 strains into 2 major groups.Based on A24A/and R27/K amino acid mutations of capsid protein,the 8 identified PCV3 strains were separated to 2 clades.This was the first detailed investigation into the epidemiology of PCV3 in Shanxi Province.Our findings enabled us to assess the possibility of widespread transmission from this region.Thus,current findings establish a basis for further studies of genetic variations in PCV3 strains circulating in China.展开更多
猪圆环病毒3型(porcine circovirus type 3,PCV3)是2016年由美国首次报道引起母猪繁殖障碍的一种新型猪圆环病毒,该病毒的致病性尚不完全清楚。为研究PCV3对断奶仔猪的致病性,本研究用PK-15细胞自临床病料分离到1株PCV3毒株,命名为PCV3/...猪圆环病毒3型(porcine circovirus type 3,PCV3)是2016年由美国首次报道引起母猪繁殖障碍的一种新型猪圆环病毒,该病毒的致病性尚不完全清楚。为研究PCV3对断奶仔猪的致病性,本研究用PK-15细胞自临床病料分离到1株PCV3毒株,命名为PCV3/CH/HB/XY/2018,全基因组测序分析表明,该毒株属于PCV3a-IM基因亚型。分别用PCV3阳性病料初始匀浆液和第8代细胞培养物采用滴鼻和肌肉注射2种方式感染28日龄断奶仔猪,每组试验猪5头,并设置空白对照组。通过观察和分析试验猪的临床表现、体温、增重、病毒血症、各组织的病毒载量和病理变化等方面评价仔猪感染试验结果。结果显示:PCV3阳性病料初始匀浆液和细胞培养物在感染断奶仔猪后均会引起仔猪出现体温上升、消瘦、皮炎和生长缓慢等现象;病毒血症时间超过14 d,但21 d消失,病毒的组织分布结果表明,PCV3主要感染扁桃体、淋巴结等免疫器官,在脾、肺和扁桃体等组织中的病毒载量最高;感染猪可出现间质性肺炎、淋巴小结增生和嗜酸性粒细胞增多等病理变化。本研究表明PCV3/CH/HB/XY/2018毒株对断奶仔猪有致病性,为PCV3的致病机制研究奠定了基础。展开更多
AIM: To investigate the genetic background of human defensin expression in type 1 and 2 diabetes.METHODS: Associations between DEFA1/DEFA3 gene copy number polymorphism and diabetes as well as between the promoter pol...AIM: To investigate the genetic background of human defensin expression in type 1 and 2 diabetes.METHODS: Associations between DEFA1/DEFA3 gene copy number polymorphism and diabetes as well as between the promoter polymorphisms of DEFB1 and diabetes were studied. The copy number variation of the DEFA1/DEFA3 genes was determined in 257 diabetic patients(117 patients with type 1 and 140 with type 2 diabetes). The control group consisted of 221 age- and gender-matched healthy blood donors. The cumulative copy numbers of the DEFA1/DEFA3 genes were detected by using quantitative PCR analysis. To evaluate the HNP 1-3(human neutrophil peptide 1-3 or α-defensin) levels in the circulation, plasma HNP 1-3 concentrations were measured by ELISA. The expression of DEFA1/A3 in peripheral leukocytes of the diabetic patients was measured by quantitative RT PCR analysis. Three SNPs of the human DEFB1(human defensin β-1) gene: DEFB1 G-20A(rs11362), DEFB1 C-44G(rs1800972) and DEFB1 G-52A(rs1799946) were genotyped by Custom TaqMan? Real Time PCR assay.RESULTS: Significant differences were observed in HNP1-3 levels between the healthy subjects and both groups of diabetic patients. The mean ± SE was 28.78 ± 4.2 ng/mL in type 1 diabetes, and 29.82 ± 5.36 ng/mL in type 2 diabetes, vs 11.94 ± 2.96 ng/mL in controls; P < 0.01 respectively. There was no significant difference between patients with type 1 and type 2 diabetes in the high plasma concentrations of HNP1-3. The highest concentrations of α-defensin were found in diabetic patients with nephropathy(49.4 ± 4.8 ng/mL), neuropathy(38.7 ± 4.8 ng/mL) or cardiovascular complications(45.6 ± 1.45 ng/L). There was no significant difference in the cumulative copy numbers of DEFA1/DEFA3 genes between controls and patients, or between patients with the two types of diabetes. Comparisons of HNP 1-3 plasma level and DEFA1/A3 copy number of the same patient did not reveal significant relationship between defensin-α levels and the gene copy numbers(r2 = 0.01). Similarly, no positive correlation was observed between the copy numbers and the mRNA expression levels of DEFA1/A3. Regarding the C-44G polymorphism of DEFB1, the GG "protective" genotype was much less frequent(1%-2%) among both groups of patients than among controls(9%).CONCLUSION: Elevated HNP1-3 levels in diabetes are independent of DEFA1/DEFA3 copy numbers, but GG genotype of C-44G SNP in DEFB1 gene may result in decreased defensin β-1 production.展开更多
基金supported by grants from the National High-Tech R&D Program (No. 2011AA100303)the National Key Technology R&D Program (No. 2011BAD19B01)the National Natural Science Foundation of China(No. 31271253)
文摘Background： The p21-activated kinase 1 （PAK1）is essential of microtubule assembly during oocyte meiotic maturation porcine oocytes. for mitosis and plays an important role in the regulatio in mice; however, little is known about its role in Result： Total p21-activated kinase 1 （PAK1） and phosphorylated PAK1 at Thr423 （PAK1^Thr423） were consistently expressed in porcine oocytes from the germinal vesicle （GV） to the second metaphase （MII） stages, but phosphorylation of histone H3 at Serr10 （H3^ser10） was only expressed after the GV stage. Immunofiuorescence analysis revealed that PAK1Thr423 and H3^ser10 colocalized on chromosomes after the GV stage. Blocking of endogenous PAK1^Thr423 by injecting a specific antibody decreased the phosphorylation level of H3^ser10; however, it had no impact on chromatin condensation, meiotic progression, cleavage rate of blastomeres or the rate of blastocyst formation. Conclusion： Phosphorylation of PAK1^Thr423 is a spontaneous activation process and the activated PAK1^Thr423 can promote the phosphorylation of H3^ser10; however, this pathway is not required for meiotic maturation of porcine oocytes or early embryonic development.
基金supported by the National Natural Science Foundation of China (31201771)the earmarked fund for the China Agriculture Research System (CARS-36)
文摘Granulosa cells（GCs） are somatic cells of ovary, the behaviors of GCs are important for ovarian function. MicroRNAs（miRNAs） are a class of endogenous 18–24 nucleotide（nt） non-coding RNAs, some of which have been shown to be important regulators of GCs function. miR-34c involved in the regulation of various biological processes and was identified to be a pro-apoptotic and anti-proliferative factor in many cell types. However, the roles of miR-34c in GCs function remain unknown. In this study, we used Annexin V-FITC and Ed U assays to demonstrate that miR-34c exerted pro-apoptotic and anti-proliferative effects in porcine GCs. Dual-luciferase reporter assays, quantitative real-time PCR（q RT-PCR） and Western blotting identified Forkhead box O3a（Fox O3a） as a direct target gene of miR-34c. The overexpression of FoxO3a rescued the phenotypic change caused by miR-34c in porcine GCs. In conclusion, miR-34c regulate the function of porcine GCs by targeting FoxO3a.
基金Supported by the National Key R&D Program of China(2016YFD0501105,2016YFD0501106)
文摘The development history, etiology, epidemiology, clinical symptoms, pathological changes and diagnosis technology of porcine circovirus 3 were summarized, and countermeasures for prevention and control of the disease were put forward based on the prevalence status in China.
文摘Porcine placenta extract (PPE) is widely accepted as an ingredient in complementary and alternative medicine and previous studies have reported its availability, however, its underlying action mechanism remains unclear. In this study, we investigated the underlying mechanism of PPE-induced ceramide synthase 3 upregulation. PPE enhanced the expression of ceramide synthase 3 at both the mRNA and protein levels in HaCaT cells. Moreover, PPE-induced ceramide synthase3 upregulation was suppressed by the Akt inhibitor, suggesting the involvement of Akt in the underlying mechanism. As the PI3K inhibitor did not affect PPE-induced ceramide synthase 3 upregulation, the factors upstream of Akt were estimated. Inhibition and small interfering RNA experiments demonstrated that the peroxisome proliferator-activated receptors δ (PPARδ) and integrin-linked kinase (ILK) are involved in the phosphorylation of Akt. Next, we explored the factors downstream of Akt and found that PPE induced phosphorylation of the STAT3 while PPE-induced upregulation of ceramide synthase 3 was significantly suppressed by the inhibitor of the mTOR, suggesting that the mTOR/STAT3 pathway is involved in the downstream of Akt. These results demonstrate that PPE upregulates CerS3 expression via the PPARδ/ILK/Akt/mTOR/STAT3 pathway.
基金This research was supported by funding from the Key Research and Development Projea Key Program of Shanxi Province,China(grant no.201703D211001).
文摘Porcine circovirus type 3(PCV3),which was first detected in the United States of America in 2015,is a potential threat to the swine industry.However,the prevalence of PCV3 in Shanxi Province,China,is unclear.In this research,the prevalence and genetic diversity of PCV3 were investigated in above area.Lung tissue samples(n=491)from 19 pig slaughterhouses across 11 cities throughout Shanxi Province were analyzed for PCV3 infection by PCR in 2019.The results showed that PCV3 positive rates in slaughterhouses and individuals were 100%(19/19)and 86.76%(426/491),respectively.PCV2 and PCV3 double-positive rates in slaughterhouses and individuals were 100%(19/19)and 59.27%(291/491),respectively.PCR positive samples were further sequenced and 8 PCV3 isolates were identified.The nucleotide homology of these isolates with other PCV3 isolates in NCBI database was 97.45-99.90%.A phylogenetic analysis,based on the complete genomic sequence and ORF2,divided these PCV3 strains into 2 major groups.Based on A24A/and R27/K amino acid mutations of capsid protein,the 8 identified PCV3 strains were separated to 2 clades.This was the first detailed investigation into the epidemiology of PCV3 in Shanxi Province.Our findings enabled us to assess the possibility of widespread transmission from this region.Thus,current findings establish a basis for further studies of genetic variations in PCV3 strains circulating in China.
基金Supported by TáMOP-4.2.2.A-11-1-KONV-2012-0035 Research Grant
文摘AIM: To investigate the genetic background of human defensin expression in type 1 and 2 diabetes.METHODS: Associations between DEFA1/DEFA3 gene copy number polymorphism and diabetes as well as between the promoter polymorphisms of DEFB1 and diabetes were studied. The copy number variation of the DEFA1/DEFA3 genes was determined in 257 diabetic patients(117 patients with type 1 and 140 with type 2 diabetes). The control group consisted of 221 age- and gender-matched healthy blood donors. The cumulative copy numbers of the DEFA1/DEFA3 genes were detected by using quantitative PCR analysis. To evaluate the HNP 1-3(human neutrophil peptide 1-3 or α-defensin) levels in the circulation, plasma HNP 1-3 concentrations were measured by ELISA. The expression of DEFA1/A3 in peripheral leukocytes of the diabetic patients was measured by quantitative RT PCR analysis. Three SNPs of the human DEFB1(human defensin β-1) gene: DEFB1 G-20A(rs11362), DEFB1 C-44G(rs1800972) and DEFB1 G-52A(rs1799946) were genotyped by Custom TaqMan? Real Time PCR assay.RESULTS: Significant differences were observed in HNP1-3 levels between the healthy subjects and both groups of diabetic patients. The mean ± SE was 28.78 ± 4.2 ng/mL in type 1 diabetes, and 29.82 ± 5.36 ng/mL in type 2 diabetes, vs 11.94 ± 2.96 ng/mL in controls; P < 0.01 respectively. There was no significant difference between patients with type 1 and type 2 diabetes in the high plasma concentrations of HNP1-3. The highest concentrations of α-defensin were found in diabetic patients with nephropathy(49.4 ± 4.8 ng/mL), neuropathy(38.7 ± 4.8 ng/mL) or cardiovascular complications(45.6 ± 1.45 ng/L). There was no significant difference in the cumulative copy numbers of DEFA1/DEFA3 genes between controls and patients, or between patients with the two types of diabetes. Comparisons of HNP 1-3 plasma level and DEFA1/A3 copy number of the same patient did not reveal significant relationship between defensin-α levels and the gene copy numbers(r2 = 0.01). Similarly, no positive correlation was observed between the copy numbers and the mRNA expression levels of DEFA1/A3. Regarding the C-44G polymorphism of DEFB1, the GG "protective" genotype was much less frequent(1%-2%) among both groups of patients than among controls(9%).CONCLUSION: Elevated HNP1-3 levels in diabetes are independent of DEFA1/DEFA3 copy numbers, but GG genotype of C-44G SNP in DEFB1 gene may result in decreased defensin β-1 production.
