硝尔库勒湖可培养放线菌多样性及其功能酶和抗细菌活性

【目的】认识和了解硝尔库勒湖可培养放线菌的多样性、功能酶和抗细菌活性特点,为今后的开发和利用奠定基础。【方法】应用可培养技术和基于16S r RNA基因序列的系统发育分析硝尔库勒盐湖沉积物中放线菌的多样性。常规方法检测样品成分因子,并筛选了嗜盐放线菌的蛋白酶、淀粉酶和酯酶活性;抑菌圈法检测放线菌新种的抗细菌活性。【结果】分离获得了51个OTUs,分属于24个不同的属,其中15个OTUs代表了放线菌新种;链霉菌属是优势菌属,占全部分离菌株数量的16.25%。硝尔库勒湖放线菌类群数量一定程度上受样品成分因子的协同影响。代表新种的菌株展示了良好的功能酶活性和抗细菌活性,其中代表链霉菌新种的菌株XHU5011不仅具有多种酶活性,而且具有强大的抗金黄葡萄球菌、耻垢分枝杆菌和荧光假单胞菌的能力,具有很好的开发潜能。【结论】硝尔库勒盐湖中存在丰富的可培养放线菌多样性,潜藏着大量的放线菌新资源,并且具有很好的功能酶和天然产物挖掘潜力。     

Exploration of interaction of canthaxanthin with human serum albumin by spectroscopic and molecular simulation methods

The interaction between the food colorant canthaxanthin (CA) and human serum albumin (HSA) in aqueous solution was explored by using fluorescence spectroscopy, three‐dimensional fluorescence spectra, synchronous fluorescence spectra, UV–vis absorbance spectroscopy, circular dichroism (CD) spectra and molecular docking methods. The thermodynamic parameters calculated from fluorescence spectra data showed that CA could result in the HSA fluorescence quenching. From the K_SV change with the temperature dependence, it was concluded that HSA fluorescence quenching triggered by CA is the static quenching and the number of binding sites is one. Furthermore, the secondary structure of HSA was changed with the addition of CA based on the results of synchronous fluorescence, three‐dimensional fluorescence and CD spectra. Hydrogen bonds and van der Waals forces played key roles in the binding process of CA with HSA, which can be obtained from negative standard enthalpy (ΔH) and negative standard entropy (ΔS). Furthermore, the conclusions were certified by molecular docking studies and the binding mode was further analyzed with Discovery Studio. These conclusions can highlight the potential of the interaction mechanism of food additives and HSA.     

High-Resolution Performance of Polycaprolactone Functionalized with Guanidinium Ionic Liquid for Gas Chromatography

This work firstly reported a new polycaprolactone based material functionalized with guanidinium ionic liquid (PCL-GIL) as the stationary phase with high resolution performance for capillary gas chromatography (GC). It is composed of polycaprolactone (PCL) and guanidinium ionic liquid (GIL) with amphiphilic conformation. The PCL-GIL capillary column coated by static method exhibited high column efficiency of 3942 plates/m and moderate polarity. As a result, the PCL-GIL column exhibited high-resolution capability. For a mixture of 27 analytes with a wide ranging polarity and outperformed the PCL-2OH and HP-35 columns, showing its advantageous separation capability for analytes of diverse types. Moreover, the PCL-GIL column showed high resolving capability for various positional isomers and cis-/trans-isomers, including alkylbenzenes, chlorobenzenes, naphthalenes, bromonitrobenzenes, chloronitrobenzenes, benzaldehydes, phenols, alcohols, respectively. In a word, PCL derivatized by GIL units as a new type of stationary phase has a promising future in GC separations.     

热环境对虎斑颈槽蛇卵孵化期、孵化成功率和孵出幼体特征的影响

用恒温（２４、２７、３０和３３Ｃ）和波动温度（平均２６．１Ｃ,范围２０．１—３２．７Ｃ）孵化虎斑颈槽蛇（Ｒｈａｂｄｏｐｈｉｓ　ｔｉｇｒｉｎｕｓ　ｌａｔｅｒａｌｉｓ）卵,检测热环境对孵化期、孵化成功率和孵出幼体的影响。孵化热环境显著影响孵化期、孵化成功率和胚胎畸形率,对孵出幼体性别无显著影响。孵化期随孵化温度升高而缩短,２４、２７、３０和３３Ｃ的平均孵化期分别为４５．０、３２．７、２７．３和２６．０ｄ,波动温度的平均孵化期为３７．９ｄ。３３Ｃ孵化成功率最低（１６．７％）,胚胎畸形率最高（１００％）。孵出幼体总性比（雌性／雄性＝０．６）不显著偏离１：１。孵出幼体的尾长显示两性异形,雄性尾长大于雌性;其它被检幼体特征无显著的两性差异。２４、２７和３０Ｃ以及波动温度孵出幼体的所有被检指标均无显著差异。３３Ｃ孵出幼体的体重和个体大小小于其它热环境中孵出的幼体,并特征性地具有较小的躯干、较大的剩余卵黄。３３Ｃ中胚胎发育的能耗显著大于其它热环境中胚胎发育的能耗。３３（‘孵出幼体的灰分含量较低,但孵出卵卵壳较重。３３Ｃ孵出幼体不能运动;其它热环境中孵出的幼体在跑道上表现良好,这些幼体的不问断运动的最大距离、每分钟运动距离和每分钟停顿次数无显著的差异。结果表明,持续将虎斑颈槽蛇卵暴露在３３Ｃ条件下不利于该种胚胎发育并可能对胚胎具有致死性影响,波动温度孵卵有利于拓宽存活孵化温度范围。     

Nitric Oxide Synthase-3 Promotes Embryonic Development of Atrioventricular Valves

Nitric oxide synthase-3 (NOS3) has recently been shown to promote endothelial-to-mesenchymal transition (EndMT) in the developing atrioventricular (AV) canal. The present study was aimed to investigate the role of NOS3 in embryonic development of AV valves. We hypothesized that NOS3 promotes embryonic development of AV valves via EndMT. To test this hypothesis, morphological and functional analysis of AV valves were performed in wild-type (WT) and NOS3^−/− mice at postnatal day 0. Our data show that the overall size and length of mitral and tricuspid valves were decreased in NOS3^−/− compared with WT mice. Echocardiographic assessment showed significant regurgitation of mitral and tricuspid valves during systole in NOS3^−/− mice. These phenotypes were all rescued by cardiac specific NOS3 overexpression. To assess EndMT, immunostaining of Snail1 was performed in the embryonic heart. Both total mesenchymal and Snail1⁺ cells in the AV cushion were decreased in NOS3^−/− compared with WT mice at E10.5 and E12.5, which was completely restored by cardiac specific NOS3 overexpression. In cultured embryonic hearts, NOS3 promoted transforming growth factor (TGFβ), bone morphogenetic protein (BMP2) and Snail1expression through cGMP. Furthermore, mesenchymal cell formation and migration from cultured AV cushion explants were decreased in the NOS3^−/− compared with WT mice. We conclude that NOS3 promotes AV valve formation during embryonic heart development and deficiency in NOS3 results in AV valve insufficiency.     

牛蛙皮胶原蛋白酸提取条件优化及其对细胞生长和粘附性的影响

应用羟脯氨酸法测定了牛蛙皮胶原蛋白含量,通过正交实验对牛蛙皮胶原蛋白酸法提取条件进行了优化,并结合MTT法测定了胶原蛋白对细胞生长和粘附性的影响。结果显示,牛蛙皮胶原蛋白的含量约为45．1％;温度条件对提取率影响最大,其次依次为酸种、时间和浓度,最优组合为乙酸、1．5mol／L、37℃、24h;在低浓度时,牛蛙皮胶原蛋白对正常人类肝细胞的生长和粘附性无显著影响,当浓度升高到一定值时,对细胞生长和粘附性有显著促进作用。     

SNHG16 as the miRNA let-7b-5p sponge facilitates the G2/M and epithelial-mesenchymal transition by regulating CDC25B and HMGA2 expression in hepatocellular carcinoma

Long noncoding RNAs (lncRNAs) play crucial roles in hepatocellular carcinoma (HCC). However, the underlying molecular mechanisms of small nucleolar RNA host gene 16 (SNHG16) for regulating the cell cycle and epithelial to mesenchymal transition (EMT) remain elusive. In this study, SNHG16 expression profiles of HCC tissues or cell lines were compared with those of normal tissues or hepatocyte cell line. The effect of SNHG16 knockdown in HCC cell lines was investigated by using in vitro loss-of-function experiments and in vivo nude mouse experiments. The potential molecular regulatory mechanism of SNHG16 in HCC progression was investigated by using mechanistic experiments and rescue assays. The results revealed that SNHG16 was highly expressed in HCC tissues and cell lines, which predicted poor prognosis of HCC patients. On one hand, the downregulation of SNHG16 induced G2/M cell cycle arrest, inducing cell apoptosis and suppression of cell proliferation. On the other hand, it inhibited cell metastasis and EMT progression demonstrated by in vitro loss-of-function cell experiments. Besides, knockdown of SNHG16 increased the sensitivity of HCC cells to cisplatin. For the detailed mechanism, SNHG16 was demonstrated to act as a let-7b-5p sponge in HCC. SNHG16 facilitated the G2/M cell cycle transition by directly acting on the let-7b-5p/CDC25B/CDK1 axis, and promoted cell metastasis and EMT progression by regulating the let-7b-5p/HMGA2 axis in HCC. In addition, the mechanism of SNHG16 for regulating HCC cell proliferation and metastasis was further confirmed in vivo by mouse experiments. Furthermore, these results can provide new insights into HCC treatment and its molecular pathogenesis, which may enlighten the further research of the molecular pathogenesis of HCC.     

隆水蚤科多样性与生态学研究进展

隆水蚤科以其丰富的个体数和多样性成为海洋水体生态系统中极为重要的小型桡足类, 但其分类学和多样性研究仍有较大不足, 其生态功能及在生态系统中的地位存在被低估的可能。为了提升对隆水蚤科的认识, 本文对国际隆水蚤科分类学和物种多样性研究进展、隆水蚤科的物种多样性研究难点和技术发展趋势、隆水蚤科的分布和生态等方面研究进行概述。19世纪末Giesbrecht创建隆水蚤科, 随后该科的新物种不断被发现和描述, 目前已描述115种。中国海仅记录到隆水蚤科物种11种, 相关生态学研究较薄弱。隆水蚤科由于个体小, 许多物种间具有高度的形态相似性, 并且包含很多姐妹种及种内分型, 因此许多研究将传统分类鉴定手段与分子生物学技术相结合, 以提高物种的发现和描述效率。随着研究的不断深入, 有关隆水蚤科物种的分布特征、食性特征、种群特征和行为学特征等均得到不同程度的关注, 这都将提高隆水蚤科研究的广度和深度。随着研究技术的迅速发展以及许多设备先进的科学考察船和载人潜水器被用于海洋研究, 从近海、边缘海到深远海研究的协同发展, 海洋生物样品资源不断丰富, 这将带动我国分类和多样性研究快速发展, 使隆水蚤科的分类学研究不断深入。     

HIV-1 gp41的酵母表面展示及表达优化

以His标签检测蛋白的表达, 利用酿酒酵母表面展示系统, 成功地将HIV-1 gp41片段锚定在酵母表面, 并检测到gp41的活性。以pMD18T-gp41为模板, 通过PCR技术克隆了gp41基因, 将gp41基因通过双酶切连接到载体pICAS-His上,构建了gp41酵母表面展示载体, 并将其转化至酿酒酵母(Saccharomyces cerevisiae)MT8-1中。重组菌经培养, 利用免疫荧光染色方法进行染色, 显微镜观察发现重组酵母细胞表面有绿色荧光, 流式细胞仪结果进一步证实gp41正确折叠展示于酵母细胞表面。采用不同浓度的葡萄糖培养基进行表达优化。当葡萄糖浓度为1%时, 82.46%的酵母细胞表达了gp41抗原; 随着葡萄糖浓度升高, 蛋白表达受到抑制。