Establishment of La-tPA/G-CSF dual transgenic mice and expression in their mammary gland

Expression vectors of human granulocyte colony stimulating factor (G-CSG) and long acting tissue plasminogen activator (La-tPA) in mammary gland were constructed using promoters of mouse whey acid protein gene (WAP) and sheep β-lactoglobulin gene (BLG) with sizes of 2.6 and 5 kb respectively. Two kinds of transgenic mice of G-CSF and La-tPA were produced with microinjection. The expression of G-CSF and La-tPA was achieved in mammary glands of transgenic mice, respectively. In order to establish dual transgenic mice of La-tPA /G-CSF, transgenic mice carrying G-CSF and La-tPA gene characterized with specific expression in mammary gland were mated. La-tPA/G-CSF dual transgenic mice were screened out from the hybrid offspring by Once-PCR. The co-expression of La-tPA and G-CSF in mammary gland of the dual transgenic mice was confirmed by the milk assayed and Northern blot analysis. Some parameters about the dual transgenic mice indicated that there were fewer litters than that of normal mice. The ratio of dual transgenes was 46.1% in F1 generation, and offspring’s sex ratio was normal. Hence a dual transgenic mouse model was established for the study of co-expression foreign proteins in mammary gland.     

Circular RNA circABCC4 as the ceRNA of miR‐1182 facilitates prostate cancer progression by promoting FOXP4 expression

In recent years, circular RNAs (circRNAs) have been identified to be essential regulators of various human cancers. However, knowledge of the functions of circRNAs in prostate cancer remains very limited. The correlation between circABCC4 and human cancer is largely unknown. This study aims to investigate the biological functions of circABCC4 in prostate cancer progression and illustrate the underlying mechanism. We found that circABCC4 was remarkably up‐regulated in prostate cancer tissues and cell lines and promoted FOXP4 expression by sponging miR‐1182 in prostate cancer cells. CircABCC4 knockdown markedly suppressed prostate cancer cell proliferation, cell‐cycle progression, migration and invasion in vitro. Furthermore, silencing of the circRNA also delayed tumor growth in vivo. Taken together, our findings indicated that circABCC4 facilitates the malignant behaviour of prostate cancer by promoting FOXP4 expression through sponging of miR‐1182. The circABCC4–miR‐1182‐FOXP4 regulatory loop may be a promising therapeutic target for prostate cancer intervention.     

Fluorescent labeling for clonal selection of Marc 145 cells secreting high levels of recombinant protein PBD-1

Despite the powerful impact gene expression markers like the green fluorescent protein (GFP) or enhanced GFP (EGFP) exert on linking the expression of recombinant protein for selection of high producers in recent years, there is still a strong incentive to develop more economical and efficient methods for isolating mammalian cell clones secreting high levels of recombinant proteins. Here we present a new method based on the co-expression of EGFP that allows clonal selection in standard 96-well cell culture plates. The genes encoding the EGFP protein and the related protein are linked by an internal ribosome entry site and thus are transcribed into the same mRNA in an independent translation process. Since both proteins arise from a common mRNA, the EGFP expression level correlates with the expression level of the therapeutic protein in each clone. By expressing recombinant porcine β-defensin 1 in Marc 145 cells, we demonstrate the robustness and performance of this technique. The method can be served as an alternative to identify high-producer clones with various cell sorting methods.     

美引烤烟品种NC102品质特征分析

通过对美引烤烟品种NC102、红花大金元和K326的化学成分、致香成分和感官评吸比较分析,进一步明晰了NC102的品质特征.NC102品种烤烟在石油醚提取物总量及新植二烯含量上具有明显优势,清甜香气风格突出,与K326比,在香气的细腻度、圆润性、飘逸感方面优于K326品种,在叶组配方中可以用作次主料使用.     

猪产仔数分子标记及其效应分析

初步确定了2个新的猪产仔数分子标记,雌激素受体基因ESR的第8外显子处的ESRB位点、催乳素受体基因的第7外显子的FSHRB位点。通过比较和分析多个产仔数的效应,初步确定了4个有利于产仔数提高的分子标记基因型,基因位点ESR、FSHRB的基因型BB的产仔数显著地高于AB、AA型;位点ESRB、PRLR的基因型AA的产仔数显著地高于AB、BB型;4个基因位点多态性与仔猪生长性能、母猪乳头数不存在显著的影响。     

山西阳泉老窑水硫酸盐还原菌的分离鉴定及其还原功能驯化

【背景】含硫煤矿开采后,地表水/地下水回流至采空区形成酸性老窑水,含有高浓度重金属离子和硫酸盐,严重危害生态系统健康。利用微生物自身生长处理老窑水具有成本低、环境友好等特点,具有良好的应用前景。目前利用的硫酸盐还原菌大多只在适宜温度和中性pH条件下具有较高活性,在北方低温和酸性条件下难以发挥作用。【目的】本研究旨在从山西阳泉山底河流域的老窑水环境中分离硫酸盐还原菌,并调节温度和pH进行驯化,从而得到高效耐低温耐酸菌株,为北方老窑水微生物治理提供可用菌种资源。【方法】对山底河流域典型老窑水样品中的微生物进行富集培养,并筛选硫酸盐还原菌。通过革兰氏染色、扫描电镜对菌株形貌特性进行表征,利用16SrRNA基因序列比对进行菌种鉴定,探究其生长特性和硫酸盐还原性能。在此基础上降低温度和pH,对高效还原硫酸盐菌株进行驯化,探讨其在北方老窑水污染治理中的应用潜力。【结果】本研究筛选得到2株硫酸盐还原菌,命名为YQ-1和YQ-2,分别属于革兰氏阴性瘤胃解蛋白质菌属(Proteiniclasticum)和脱硫弧菌属(Desulfovibrio)。在30°C、pH 7.5条件下,YQ-1和YQ-2对1 1...     

Involvement of SSAO-mediated deamination in adipose glucose transport and weight gain in obese diabetic KKAy mice

Semicarbazide-sensitive amine oxidase (SSAO) is located on outer surfaces of adipocytes and endothelial and vascular smooth muscle cells. This enzyme catalyzes deamination of methylamine and aminoacetone, leading to production of toxic formaldehyde and methylglyoxal, respectively, as well as hydrogen peroxide and ammonium. Several lines of evidence suggest that increased SSAO activity is related to chronic inflammation and vascular disorders related to diabetic complications. We found that a highly potent and selective SSAO inhibitor, (E)-2-(4-fluorophenethyl)-3-fluoroallylamine (FPFA), was capable of reducing numbers of atherosclerotic lesions as well as weight gain in obese KKAy mice fed an atherogenic diet. SSAO inhibitors cause a moderate and long-lasting hyperglycemia. Such an increase in serum glucose is a result of reduction of glucose uptake by adipocytes. SSAO-mediated deamination of endogenous methylamine substrates induces adipocyte glucose uptake and lipogenesis. Highly selective SSAO inhibitors can effectively block induced glucose uptake. The results suggest that increased SSAO-mediated deamination may be concomitantly related to obesity and vascular disorders associated with type 2 diabetes.     

沙枣花蜜腺的发育解剖学研究

沙枣的花蜜腺位于花柱基部的筒状花盘上,属花盘蜜腺,其蜜腺位于花盘外方,由分履表皮和产蜜组织组成。分泌表皮具有角质层和变态的气孔器。产蜜组织在发育过程中,其液泡和淀粉粒都随着蜜腺的发育呈现一定的消长规律,最后形成的蜜汁由盘状蜜腺表面的气孔泌出。     

西藏牦牛血液酶活性与生产性能的相关性研究

测定了 2 8头西藏牦牛血液中 6种酶的活性 ,探讨了 6种酶活性与生产性能的关系。相关分析表明 ,LDH活性与产奶量、CAT活性与腹毛长呈极显著的正相关 (P <0 .0 1 ) ,AKP活性与体重、Amy活性与体重呈显著正相关 (P <0 .0 5 ) ,SOD活性与体重呈显著负相关 (P <0 .0 5 )。逐步回归分析表明 ,可以用LDH活性预测牦牛产奶量 ,用AKP、CAT、Amy 3种酶活性预测牦牛体重 ,用CAT、Amy活性预测牦牛腹毛长。因此 ,有望将LDH、AKP、CAT和Amy等酶活性作为生化遗传标记应用于牦牛产奶量、体重、腹毛长的选择中     

Repression of SIRT1 Promotes the Differentiation of Mouse Induced Pluripotent Stem Cells into Neural Stem Cells

The use of transplanting functional neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) has increased for the treatment of brain diseases. As such, it is important to understand the molecular mechanisms that promote NSCs differentiation of iPSCs for future NSC-based therapies. Sirtuin 1 (SIRT1), a NAD⁺-dependent protein deacetylase, has attracted significant attention over the past decade due to its prominent role in processes including organ development, longevity, and cancer. However, it remains unclear whether SIRT1 plays a role in the differentiation of mouse iPSCs toward NSCs. In this study, we produced NSCs from mouse iPSCs using serum-free medium supplemented with retinoic acid. We then assessed changes in the expression of SIRT1 and microRNA-34a, which regulates SIRT1 expression. Moreover, we used a SIRT1 inhibitor to investigate the role of SIRT1 in NSCs differentiation of iPSCs. Data revealed that the expression of SIRT1 decreased, whereas miRNAs-34a increased, during this process. In addition, the inhibition of SIRT1 enhanced the generation of NSCs and mature neurocytes. This suggests that SIRT1 negatively regulated the differentiation of mouse iPSCs into NSCs, and that this process may be regulated by miRNA-34a.