小鼠原核期受精卵体外培养系统的筛选

为了筛选出最佳的小鼠原核期受精卵的体外发育培养系统,分别进行了四个试验。试验I:在体外分别用自配的M16、mM16、KSOM、mKSOM、CZB进行体外发育培养,进而筛选出一种最佳的体外培养系统;试验Ⅱ、Ⅲ、Ⅳ分别:探讨了血清、PVA、rhLIF对小鼠胚胎发育的影响。结果,试验I中胚胎发育到2-细胞的比率差异不明显,但是在mM16和mKSOM中,发育到4-细胞的比率94.7%,90.7%(91/96;78/86)和发育到桑椹胚/囊胚的比率分别为78.1%,67.4%(75/96;58/86)均明显高于其他三种培养液;试验II用10?S代替M16中BSA时,胚胎的发育率均下降,即使在mM16中桑椹胚/囊胚率仅为4.8%(12/35)与对照组M16(40.5%)差异显著(p<0.05);试验Ⅲ用PVA取代mM16和mKSOM中的BSA其体外发育率显著下降,胚胎均无一例发育到桑椹胚/囊胚;试验Ⅳ:rhLIF能提高胚胎在体外的发育率可使mM16培养的胚胎囊胚率、囊胚脱出率分别达到84%(47/56)、39.2%(22/56)。结论:在不添减其他成分前提下,只在M16中添加0.1mMolEDTA、0.5mMol牛磺酸、1000IU/mlrhLIF便可获得84%的囊胚率,同时证明在M16或mM16添加血清都会降低其体外发育率;PVA还不能有效的取代mM16、mKSOM中的血清。     

The Regulation and Function of Histone Methylation

Histones are wrapped around by genomic DNA to form nucleosomes which are the basic units of chromatin. In eukaryotes histones undergo various covalent modifications such as methylation, phosphorylation, acetylation, ubiquitination and ribosylation. Histone modifications play a fundamental role in the epigenetic regulation of gene expression in multicellular eukaryotes. Histone methylation is one of the most important modifications occurring on Lysine (K) and Arginine (R) residues of histones, dynamically regulated by histone methyltransferases and demethylases. Identifications of such histone modification enzymes and to study how they work are the most fundamental questions needs to be answered. Uncovering the regulation and functions of the various histone methylation enzymes will help us to better understand the epigenetic code. This review summarizes the regulation of histone methyltransferases activity, the recruitment of methyltransferases and the distribution patterns and function of histone methylations.     

CO2倍增对不同基因型大豆光合色素含量和荧光诱导动力学参数的影响

研究了CO_2倍增对大豆(Glycine max L.)Bragg(野生型)及其不同单基因突变品系Nts 382(超结瘤突变体)和Nod 49(不结瘤突变体)某些光合特性的影响。结果表明,CO_2倍增能提高Bragg、Nts 382和Nod 49的叶绿素(Chl)和类胡萝卜素(Car)的含量,但不同品系提高的幅度有所不同。荧光诱导动力学测定结果表明,CO_2倍增均能提高其PSⅡ活性、PSⅡ原初光能转化效率和光合作用潜在量子转化效率。CO_2倍增更有利于提高Nts 382的荧光光化学猝灭系数(qp)和PSⅡ总的光化学量子产量,以及较大幅度地降低荧光非光化学猝灭系数(qN),有助于把所捕获的光能用于进行光合作用。这可能与Nts 382是超结瘤突变体,比Bragg和Nod 49能更充分地利用空气中的氮素有关。     

辽东山区天然次生林的数量分类

结合DCA排序和TW IN SPAN分类结果,将辽东山区天然次生林划分为5个群落类型:花曲柳林、蒙古栎林、阔叶混交林、水曲柳林、胡桃楸林。DCA排序与TW IN SPAN分类产生了较一致的分类结果。DCA第一轴代表的环境意义为坡向。花曲柳林多分布于阳坡的中坡及中上坡,蒙古栎林多分布在中上坡。花曲柳与蒙古栎常混生在一起,随着坡位上升,花曲柳优势度下降,而蒙古栎优势度增加。阔叶混交林多位于阴坡,乔木层没有稳定和绝对优势种,多以假色槭、风桦、色木槭为优势种,但假色槭分布于乔木层第2亚层。水曲柳林多分布在中下坡。胡桃楸分布在山下部,喜潮湿生境。色木槭在辽东山区分布广泛,重要值较高,且更新良好,很可能是群落演替顶级物种。辽东山区天然次生林林下藤本植株数量多,以五味子、软枣猕猴桃、狗枣猕猴桃为主。     

基于代谢通量分析的琥珀酸放线杆菌高产选育研究

在对产琥珀酸放线杆菌代谢分析的基础上选育出高产突变株对琥珀酸的工业生物转化有重要意义.在矩阵分析代谢通量基础上,围绕柔性节点下的副产物乙酸及乙醇的降低分别实施软X诱变及定点突变选育,并对比分析了突变株与出发株相关酶活及基因序列变化.针对出发株的流量分析显示产物琥珀酸的代谢通量为1.78(mmol/g/h),主要副产物乙酸与乙醇的代谢通量分别为(0.60mmol/g/h)和(1.04 mmol/g/h),并发现乙醇代谢加剧了琥珀酸合成中的H电子供体的不足;筛选出的氟乙酸抗性突变株S.JST1的乙酸代谢通量降低了96%,为0.024(mmol/g/h),酶活检测表明磷酸乙酰转移酶(Pta)的酶比活力从602降低到74,进一步的序列对比分析发现pta突变基因中产生了一个突变位点:adh定点复合突变株S.JST2的乙醇代谢通量降低了98%,为0.020(mmol/g/h),酶活检测表明Adh的酶比活力从585降低到62.最终突变株S.JST2琥珀酸累积产量达65.7 g/L.围绕产琥珀酸放线杆菌Pta及Adh酶活的降低实施定向选育,在降低副产物流量的同时,有助于改善细胞H供体代谢平衡进而提高琥珀酸的流量.所获突变株具有工业应用潜力.     

Construction of a cross-species cell landscape at single-cell level

Individual cells are basic units of life. Despite extensive efforts to characterize the cellular heterogeneity of different organisms, cross-species comparisons of landscape dynamics have not been achieved. Here, we applied single-cell RNA sequencing (scRNA-seq) to map organism-level cell landscapes at multiple life stages for mice, zebrafish and Drosophila. By integrating the comprehensive dataset of > 2.6 million single cells, we constructed a cross-species cell landscape and identified signatures and common pathways that changed throughout the life span. We identified structural inflammation and mitochondrial dysfunction as the most common hallmarks of organism aging, and found that pharmacological activation of mitochondrial metabolism alleviated aging phenotypes in mice. The cross-species cell landscape with other published datasets were stored in an integrated online portal—Cell Landscape. Our work provides a valuable resource for studying lineage development, maturation and aging.     

Intra‐amniotic mesenchymal stem cell therapy improves the amniotic fluid microenvironment in rat spina bifida aperta fetuses

ObjectivesSpina bifida aperta (SBA) is one of the most common neural tube defects. Neural injury in SBA occurs in two stages involving failed neural tube closure and progressive degeneration through contact with the amniotic fluid. We previously suggested that intra‐amniotic bone marrow‐derived mesenchymal stem cell (BMSC) therapy for fetal rat SBA could achieve beneficial functional recovery through lesion‐specific differentiation. The aim of this study is to examine whether the amniotic fluid microenvironment can be improved by intra‐amniotic BMSC transplantation.MethodsThe intra‐amniotic BMSC injection was performed using in vivo rat fetal SBA models. The various cytokine expressions in rat amniotic fluid were screened by protein microassays. Intervention experiments were used to study the function of differentially expressed cytokines.ResultsA total of 32 cytokines showed significant upregulated expression in the BMSC‐injected amniotic fluid. We focused on Activin A, NGF, BDNF, CNTF, and CXCR4. Intervention experiments showed that the upregulated Activin A, NGF, BDNF, and CNTF could inhibit apoptosis and promote synaptic development in fetal spinal cords. Inhibiting the activity of these factors weakened the anti‐apoptotic and pro‐differentiation effects of transplanted BMSCs. Inhibition of CXCR4 activity reduced the engraftment rate of BMSCs in SBA fetuses.ConclusionBMSC transplantation can improve the amniotic fluid environment, and this is beneficial for SBA repair.

In utero intra‐amniotic BMSC or PBS microinjection in the E15 fetuses was performed in E15 rat fetuses with spina bifida aperta, and amniotic fluid was collected at E21 for protein array detection. Venn diagram shows the relationship of three biological processes (GO: 0030335, 0048699, and 0043524) and the attribution of differentially expressed proteins. Comparative analysis of five proteins with the largest fold changes in the process of generation of neurons.     

Discovery of novel mRNA demethylase FTO inhibitors against esophageal cancer

A series of 1,2,3-triazole analogues as novel fat mass and obesity-associated protein (FTO) inhibitors were synthesised in this study. Among all 1,2,3-triazoles, compound C6 exhibited the most robust inhibition of FTO with an IC₅₀ value of 780 nM. It displayed the potent antiproliferative activity against KYSE-150, KYSE-270, TE-1, KYSE-510, and EC109 cell lines with IC₅₀ value of 2.17, 1.35, 0.95, 4.15, and 0.83 μM, respectively. In addition, C6 arrested the cell cycle at G2 phase against TE-1 and EC109 cells in a concentration-dependent manner. Analysis of cellular mechanisms demonstrated that C6 concentration-dependently regulated epithelial mesenchymal transition (EMT) pathway and PI3K/AKT pathway against TE-1 and EC109 cells. Molecular docking studies that C6 formed important hydrogen-bond interaction with Lys107, Asn110, Tyr108, and Leu109 of FTO. These findings suggested that C6 as a novel FTO inhibitor and orally antitumor agent deserves further investigation to treat esophageal cancer.