Blastocyst morphology, actin cytoskeleton quality and chromosome content are correlated with embryo quality in the pig

Embryo survival rates obtained after transfer of in vitro produced porcine blastocysts are very poor. This is probably related to poor quality of the embryos. The aim of the present study was to determine markers for good quality blastocysts. Therefore, we tried to link blastocyst morphology to several morphological and cell biological properties, and evaluated the survival of in vitro produced, morphologically classified, blastocysts following non-surgical transfer. In vitro and in vivo produced blastocysts were allocated to two groups (classes A and B) on the basis of morphological characteristics. The quality of their actin cytoskeleton, their total cell number, their ability to re-expand after cytochalasin-B treatment and the occurrence of numerical chromosome aberrations were studied and compared. In vivo produced blastocysts were used as a control. Our results indicate that the ability of blastocysts to re-expand after cytochalasin-B-induced actin depolymerization was positively correlated with the morphology of the blastocyst, and associated with the quality of the actin cytoskeleton. Chromosome analysis revealed that mosaicism is inherent to the in vitro production of porcine embryos, but also that in vivo produced blastocysts contained some non-diploid cells. In non-surgical embryo transfer experiments more recipients receiving class A blastocysts were pregnant on Day 20 than those receiving class B blastocysts. One recipient gave birth to six piglets from class A in vitro produced blastocysts, providing a verification of the enhanced viability of blastocysts that were scored as 'good' on the basis of their morphology.     

三种杜拉属（Drawida）蚯蚓的酯酶同工酶的初步研究

应用聚丙烯酰胺凝胶电泳的方法，对寡毛纲杜拉属三种蚯蚓进行酯酶同工酶酶谱分析。其结果表明，同属蚯蚓具有几条相似的酶谱区带，而不同种蚯蚓又具有各自独特的酶谱带，种间酶谱区带数目、泳动率及染色强度都有明显区别。雅和杜拉蚓酶谱图和天锡杜拉蚓相似，而与管状杜拉蚓相差较远，表明同属蚯蚓在进化水平上的亲缘关系。     

研究报告：CRBGP通过抑制FAK-AKT通路和白介素-6的分泌抑制胶质瘤U251细胞的活性

目的电压门控钠通道(voltage-gated sodium channels,VGSCs)表达于胶质瘤U251细胞,并影响U251细胞的增殖、侵袭和凋亡。富含半胱氨酸的颊腺蛋白(cysteine-rich buccal gland protein,CRBGP)是一种从日本七鳃鳗颊腺中分离出来的VGSCs阻断剂。本文旨在探究CRBGP是否因具有VGSCs阻断功能从而能够抑制U251细胞的活性。方法首先采用MTT法检测了CRBGP对U251细胞增殖的作用,并分别利用莱特-吉姆萨、FITC-phalloidin和Hoechst 33258染色法观察CRBGP处理后U251细胞的形态、细胞骨架和细胞核。细胞外基质蛋白如IV型胶原蛋白、纤连蛋白和层黏连蛋白用于检测CRBGP对U251细胞黏附的影响。此外,本文采用transwell法检测CRBGP处理后U251细胞的迁移和侵袭能力,并通过Western blot方法探讨CRBGP诱导U251细胞凋亡并抑制其运动的内在机理。最后,通过酶联免疫吸附测定法检测CRBGP对U251细胞的抑炎效果。结果 CRBGP通过线粒体依赖途径诱导U251细胞凋亡,...     

A mRNA landscape of bovine embryos after standard and MAPK-inhibited culture conditions: a comparative analysis

Background

Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed.

Results

Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated.

Conclusion

The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1448-x) contains supplementary material, which is available to authorized users.     

中国蓼属萹蓄组植物果实形态的研究

应用解剖镜和电子扫描显微镜对中国产蓼属萹蓄组Polygonum sect.Polygonum 16种1变种及1变型植物的果实形态进行了研究。发现该组5种一年生植物的果实具有两型性,即有长、短两种果实类型。短果类型为卵形,具三棱,包于宿存花被内或与花被近等长,呈黑褐色。在解剖镜下观察到其表面具小点、无光泽,而在电子扫描显微镜下观察到其表面具有散乱分布或排列成行的瘤状颗粒;长果类型为狭卵形,具三棱,长于宿存花被,黄褐色或淡黄绿色,在解剖镜下观察到其表面平滑、有光泽,而在电子扫描显微镜下观察到其表面上部具有散乱分布的瘤状颗粒,中、下部则具不规则洼点、星花状褶皱或浅波纹状。果实具两型性的植物有萹蓄P.aviculare、乌鲁木齐萹蓄P.urumqiense、展枝萹蓄P.patulum、直立萹蓄P.avicularef.erectum和尖果萹蓄P.rigidum。果实微形态可分为两种类型：1.果实表面无瘤状颗粒或疣状突起、有光泽,此类型又可以分为2种亚类型：（1）果实表面平滑、浅波纹状,如岩萹蓄P.cognatum、松叶萹蓄P.acerosum和帚萹蓄P.argyrocoleum;（2）果实表面具浅洼点或洼点,如铁马鞭P.plebeium和针叶萹蓄P.polycnemoides。2.果实表面有瘤状颗粒或疣状突起,无或具微光泽,此类型又可分为4种亚类型：（1）果实表面具疣状突起,如圆叶萹蓄P.intramongolicum.;（2）果实表面具散乱分布的瘤状颗粒,如普通萹蓄P.humifusum和展枝萹蓄的短果;（3）果实表面具有纵行排列的瘤状颗粒,如萹蓄和乌鲁木齐萹蓄的短果;（4）果实表面上部具散乱分布的瘤状颗粒,中、下部具不规则洼点、星花状褶皱或浅波纹状,如萹蓄、展枝萹蓄、直立萹蓄和尖果萹蓄的长果。因此果实两型性和果实微形态特征对蓼属萹蓄组种、变种的鉴定具有较大的分类学价值。基于果实微形态特征以及其他形态特征,确定了新种晖春萹蓄Polygonum huichunense F.Z.Li,Y.T.Hou＆C.Y.Qu,同时恢复褐鞘萹蓄Polygonum aviculareL.var.fusco-ochreatum（Kom.）A.J.Li为原种,即Polygonum fusco-ochreatum Kom.。     

2004—2018年青海省乙型病毒性肝炎流行病学特征分析

目的 了解青海省2004—2018年乙型病毒性肝炎(简称乙肝)流行病学特征,为乙肝防治工作提供科学依据。方法 对青海省15年间乙肝监测数据进行描述性流行病学分析。结果 15年来青海省共报告乙肝患者201931例,年平均报告发病率239.71/10万,总体呈下降趋势。2004—2018年西宁市乙肝报告病例98218例,占48.64%;果洛藏族自治州最低,乙肝报告病例6766例,仅占3.35%。玉树藏族自治州乙肝报告发病率最高332.89/10万,海东市最低133.43/10万,不同地区乙肝报告发病率有统计学意义(χ^2=21997.32,P<0.05)。报告病例主要分布在25~55岁人群中,50~<55岁组发病率最高为454.99/10万,0~<5岁组发病率最低为23.69/10万;男、女性别比为1.57∶1。报告病例职业分布以农、牧民为主,占47.19%。结论 青海省经过多年的计划免疫的实施,乙肝防控效果明显,2009年以后乙肝报告病例呈现平稳状况。     

蒙药材刺柏叶中槲皮苷的含量测定

本实验建立了蒙药材刺柏叶中槲皮苷的HPLC含量测定方法.色谱条件为:Diamonsil C18(4.6 mm ×250 mm,5μm)和Shim-Pack C18(4.6 mm× 250 mm,5μm),流动相:甲醇-0.01 mol/L磷酸二氢钾溶液-冰醋酸(40∶ 60∶ 1.5),检测波长:254 nm,流速:1.0 mL/min,柱温:30℃.槲皮苷进样量在40.2 ng ～603.0 ng范围内线性关系良好(r =0.9998),其平均加样回收率为100.45％,RSD为0.69％(n=6).本方法简便易行,结果准确,重复性好,可用于刺柏叶中槲皮苷的含量测定.     

新疆甘草化学成分研究

从新疆甘草(GlycyrrhizauralensisFisch.)根中分离得到5个化合物,分别鉴定为:槲皮素(1)、甘草次酸(2)、阿魏酸(3)、5,7,3′,4′-四羟基-7-葡萄糖基二氢黄酮醇(4)、8,5′-二羟基3′α-鼠李糖黄酮(5)。其中化合物4,5为首次从该植物中分离得到。     

冬葵果多糖的抗氧化作用研究

研究冬葵果多糖(Fructus Malvae polysaccharides,FMP)对氧自由基的清除作用及对脂质过氧化的抑制作用。采用分光光度法测定FMP清除Fenton体系产生的羟自由基(.OH)、邻苯三酚自氧化体系产生的超氧阴离子自由基(O2-.)的能力及评价对Fe(Ⅱ)-H2O2体系(.OH)诱导的脂质过氧化终产物丙二醛(MDA)的抑制作用。结果显示,在化学模拟体系中,冬葵果多糖对.OH具有很强的清除作用,与VC比较达到显著水平(P<0.01),对O2-.的清除能力与VC相当。在体外,多糖浓度达到1.72 mg/mL可明显降低MDA的含量,与空白液比较达到显著水平(P<0.01)。以上结果表明,FMP具有明显的抗氧化活性。