Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embr...Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.展开更多
虚拟化技术日益成为关注的焦点,正在最大程度地改变企业 IT 基础设施的部署及运营。甘肃电力协同办公系统打破原有本部、各地市公司两级部署模式,采用大集中部署服务器的技术架构构建电力业务应用系统。 2 台 IBM p6 570 小型机采用逻...虚拟化技术日益成为关注的焦点,正在最大程度地改变企业 IT 基础设施的部署及运营。甘肃电力协同办公系统打破原有本部、各地市公司两级部署模式,采用大集中部署服务器的技术架构构建电力业务应用系统。 2 台 IBM p6 570 小型机采用逻辑分区技术(LPAR),将单台服务器划分成多个逻辑服务器,彼此运行独立的应用程序。逻辑分区技术在甘肃电力协同办公系统中的应用,不仅可以减少服务器的数量,通过降低空间、散热以及电力消耗等途径提供一种服务器整合的方法,节省成本投资;而且可以提高可用性,带来具有透明负载均衡、动态迁移、故障自动隔离、系统自动重构的高可靠服务器应用环境。展开更多
目的探讨miR-15b靶向LPAR3调控食管癌细胞增殖、迁移及侵袭的影响。方法取对数生长期的人食管癌Eca109细胞分为对照组、miR-15b mimic组、miR-NC组、miR-15b mimic+pcDNA3.1组、miR-15b mimic+pcDNA-LPAR3组;另设对数期生长的人食管上...目的探讨miR-15b靶向LPAR3调控食管癌细胞增殖、迁移及侵袭的影响。方法取对数生长期的人食管癌Eca109细胞分为对照组、miR-15b mimic组、miR-NC组、miR-15b mimic+pcDNA3.1组、miR-15b mimic+pcDNA-LPAR3组;另设对数期生长的人食管上皮细胞HEEpiC为空白组。采用qRT-PCR检测各组细胞中miR-15b和LPAR3 mRNA的表达;应用MTT法、划痕实验和Transwell小室实验分别检测各组细胞的增殖、迁移和侵袭情况;Western blot检测Eca109细胞中LPAR3和增殖、迁移、侵袭相关蛋白(Cyclin D1、MMP-2、MMP-9)的表达。结果与空白组相比,对照组Eca109细胞中miR-15b水平明显降低(1.00±0 vs 0.42±0.03,t=33.486,P<0.01),LPAR3 mRNA水平显著升高(1.00±0 vs 2.33±0.15,t=15.358,P<0.01);与对照组相比,miR-15b mimic组细胞miR-15b水平显著升高(0.42±0.03 vs 4.61±0.38,t=19.039,P<0.01),LPAR3 mRNA(2.33±0.15 vs 1.29±0.13)和蛋白(1.12±0.13 vs 0.35±0.03)水平、细胞增殖率(100.00±0 vs 64.57±6.11)、划痕愈合率(63.45±7.20 vs 36.52±4.29)、Eca109细胞进入Transwell小室下层的数量(155.31±14.23 vs 101.33±11.45)、Cyclin D1(0.85±0.10 vs 0.38±0.05)、MMP-2(0.76±0.08 vs 0.33±0.04)、MMP-9(0.94±0.10 vs 0.41±0.05)蛋白表达均显著降低(t=9.075、9.996、10.044、5.565、5.119、7.281、8.327、8.211,P<0.01);使用pcDNA-LPAR3过表达LPAR3能明显逆转miR-15b mimic对细胞增殖、迁移和侵袭的抑制能力。结论食管癌中过表达miR-15b可能通过下调LPAR3表达,抑制食管癌细胞增殖、迁移和侵袭。展开更多
基金financially supported by the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(2020ZD0007)the Major Program of the Inner Mongolia Natural Science Foundation,China(2020ZD10)+3 种基金the National Natural Science Foundation of China(32160172)the Natural Science Foundation of Inner Mongolia Autonomous Region(2020BS03003 and 2020BS03022)the National Transgenic Project of China(2016ZX0801000-002 and 2016ZX08010005-001)the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(zdzx2018065)。
文摘Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.
文摘虚拟化技术日益成为关注的焦点,正在最大程度地改变企业 IT 基础设施的部署及运营。甘肃电力协同办公系统打破原有本部、各地市公司两级部署模式,采用大集中部署服务器的技术架构构建电力业务应用系统。 2 台 IBM p6 570 小型机采用逻辑分区技术(LPAR),将单台服务器划分成多个逻辑服务器,彼此运行独立的应用程序。逻辑分区技术在甘肃电力协同办公系统中的应用,不仅可以减少服务器的数量,通过降低空间、散热以及电力消耗等途径提供一种服务器整合的方法,节省成本投资;而且可以提高可用性,带来具有透明负载均衡、动态迁移、故障自动隔离、系统自动重构的高可靠服务器应用环境。
文摘目的探讨miR-15b靶向LPAR3调控食管癌细胞增殖、迁移及侵袭的影响。方法取对数生长期的人食管癌Eca109细胞分为对照组、miR-15b mimic组、miR-NC组、miR-15b mimic+pcDNA3.1组、miR-15b mimic+pcDNA-LPAR3组;另设对数期生长的人食管上皮细胞HEEpiC为空白组。采用qRT-PCR检测各组细胞中miR-15b和LPAR3 mRNA的表达;应用MTT法、划痕实验和Transwell小室实验分别检测各组细胞的增殖、迁移和侵袭情况;Western blot检测Eca109细胞中LPAR3和增殖、迁移、侵袭相关蛋白(Cyclin D1、MMP-2、MMP-9)的表达。结果与空白组相比,对照组Eca109细胞中miR-15b水平明显降低(1.00±0 vs 0.42±0.03,t=33.486,P<0.01),LPAR3 mRNA水平显著升高(1.00±0 vs 2.33±0.15,t=15.358,P<0.01);与对照组相比,miR-15b mimic组细胞miR-15b水平显著升高(0.42±0.03 vs 4.61±0.38,t=19.039,P<0.01),LPAR3 mRNA(2.33±0.15 vs 1.29±0.13)和蛋白(1.12±0.13 vs 0.35±0.03)水平、细胞增殖率(100.00±0 vs 64.57±6.11)、划痕愈合率(63.45±7.20 vs 36.52±4.29)、Eca109细胞进入Transwell小室下层的数量(155.31±14.23 vs 101.33±11.45)、Cyclin D1(0.85±0.10 vs 0.38±0.05)、MMP-2(0.76±0.08 vs 0.33±0.04)、MMP-9(0.94±0.10 vs 0.41±0.05)蛋白表达均显著降低(t=9.075、9.996、10.044、5.565、5.119、7.281、8.327、8.211,P<0.01);使用pcDNA-LPAR3过表达LPAR3能明显逆转miR-15b mimic对细胞增殖、迁移和侵袭的抑制能力。结论食管癌中过表达miR-15b可能通过下调LPAR3表达,抑制食管癌细胞增殖、迁移和侵袭。
