大熊猫精液品质的研究：精子和非精子的细胞成分超微结构观察

本文对刚接近性成熟大熊猫的精液在电子显微镜下进行精液品质鉴定,观察结果,除少量完成变态的精子外,多数是处在精子发生的各阶段并发育不正常,精液中伴随着大量巨噬细胞和被吞噬的衰亡精子及其碎片。镜检结果不仅说明这只雄性大熊猫性尚未成熟,其生殖系统可能还伴有生理或病理现象,这样的精液建议不能用于配种。     

Changes of Con A Receptor Sites on Mammalian Sperms during Capacitation and Acrosome Reaction

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老化处理对大豆种子活力及线粒体抗坏血酸-谷胱甘肽循环的影响

以大豆‘中黄13’为材料,研究人工老化后大豆种子活力及线粒体抗坏血酸-谷胱甘肽（ASC-GSH）循环的变化。结果表明,随老化时间的延长,大豆种子的发芽率、发芽势、发芽指数和活力指数均显著下降;老化种子的相对电导率和丙二醛（MDA）含量随着老化时间的延长逐渐增大,超氧阴离子（O₂?^-）产生速率和过氧化氢（H₂O₂）含量呈现先升高后降低的趋势;与对照相比,老化种子中线粒体细胞色素c氧化酶（COX）和苹果酸脱氢酶（MDH）活性显著下降,呼吸速率和呼吸控制率（RCR）显著降低;老化种子中线粒体超氧化物歧化酶（SOD）、抗坏血酸过氧化物酶（APX）、单脱氢抗坏血酸还原酶（MDHAR）、脱氢抗坏血酸还原酶（DHAR）和谷胱甘肽还原酶（GR）的活性以及总ASC和GSH含量显著降低,说明老化导致活性氧（ROS）代谢异常,线粒体呼吸功能及ASC-GSH循环紊乱。ROS的过量积累可能是导致种子活力丧失的主要原因。     

<Emphasis Type="Italic">Sus scrofa</Emphasis> matrix attachment region enhances expression of the PiggyBac system transfected into HEK293T cells

Objectives

To determine the effects of the Sus scrofa matrix attachment region (SusMAR) on transgene expression in HEK293T cells.

Results

Three expression vectors with the MAR at different sites in the PiggyBac (PB) transposon vector backbone were compared: two MARs flanking the β-galactosidase (β-gal) expression cassette, and one at the upstream or downstream site. Bos taurus MAR (BosMAR) and a β-gal expression cassette without MARs were the positive and negative controls, respectively. Compared to the control, β-gal activity of all SusMAR and BosMAR vectors was significantly improved in the presence of PB transposase (PBase). However, only the downstream SusMAR and upstream BosMAR vectors showed increased expression in the absence of PBase. Expression was significantly increased in all vectors with the PBase group compared to those without the PBase group. Gene copy numbers were not increased compared to the negative control.

Conclusions

SusMAR enhanced recombinant gene expression levels and stability in HEK293T cells, was not increase transgene copy number. These results could facilitate the development of vectors for stable production of therapeutic proteins.

     

鹞落坪国家级自然保护区吊罐井物种分布格局与种间关联的多尺度分析

为了探讨群落结构形成机制,采用点格局分析方法,对鹞落坪国家级自然保护区吊罐井核心区1hm2样地内36个优势物种在0~50m尺度上的空间分布格局和种间关联性进行了研究。结果显示,36个物种的空间分布格局在小尺度上主要呈聚集分布,在较大尺度上呈随机分布的物种比例最高;对36个物种种间关联性的分析发现,无关联类型是该样地物种间相互作用的主要表现形式,并且在10m尺度上无关联种对的比例最低,正相关和负相关的种对在所有尺度上的比例均未超过20%。群落结构的次生性、生境异质性和扩散限制性是导致该样地物种分布和种间关系格局的主要原因。本研究结果有助于了解物种共存的尺度依赖性特征及其形成机制。     

玉米酵母双杂交cDNA文库的构建及ZmCEN互作蛋白的筛选

为阐明玉米中心蛋白(ZmCEN)的生物学功能,采用酵母双杂交技术对其互作蛋白进行研究。提取玉米(Zea mays L.)自交系‘郑58’幼苗的总RNA,利用SMART技术反转录合成ds cDNA,构建以pGBKT7为载体的酵母双杂交cDNA文库;依据ZmCEN基因的CDS序列设计引物,构建重组诱饵载体(pGBKT7-ZmCEN)转化酵母菌株Y2HGold,检测诱饵载体的毒性与自激活能力后,筛选与玉米中心蛋白(ZmCEN)互作的猎物蛋白。将筛选的互作蛋白NAC67和TONNEAU1b(TON1b)再次验证相互作用,并选取互作蛋白TON1b,采用BiFC实验分别构建ZmCEN-pSPYNE和TON1b-pSPYCE BiFC半分子重组载体,转化拟南芥原生质体,进一步验证它们在细胞内的互作;并利用Uniprot和KEGG在线网站对互作蛋白进行gene ontology(GO)注释分析。结果表明:玉米全株幼苗的cDNA文库库容量达到2.56×107 CFU,文库滴度5.36×108 CFU/mL,符合建库要求。经检测诱饵载体无毒性也无自激活功能,所筛选的cDNA文库经测序和Blast比对分析以及共转验证,最终得到28个与诱饵蛋白ZmCEN互作的蛋白质。GO注释显示互作蛋白参与的生物过程有21种。BiFC结果显示,蛋白TON1b与ZmCEN在拟南芥原生质体细胞内互作而形成互补,从而产生黄色荧光,进一步证实了两者存在互作关系。酵母双杂交系统cDNA文库的成功构建与筛选,为进一步研究玉米ZmCEN及其与互作蛋白的作用机制奠定了基础。     

不同时期干旱胁迫对甘薯光合效率和耗水特性的影响

在2014-2015年遮雨棚下种植甘薯品种‘济薯21’,以全生育期正常灌水(WW)为对照,研究了全生育期(DS)、发根分枝期(DS₁)、蔓薯并长期(DS₂)和快速膨大期(DS₃)干旱胁迫对甘薯光合作用、产量和耗水特性的影响.结果表明: DS、DS₁、DS₂和DS₃的生物产量分别比WW降低31.3%、21.2%、19.6%和7.7%,收获指数分别降低19.9%、14.5%、14.1%和6.5%,薯干产量分别降低45.3%、33.1%、31.3%和14.2%.栽后100 d,DS、DS₁、DS₂和DS₃的叶面积系数分别比WW减少77.1%、60.1%、39.2%和17.1%;栽后90 d,叶片光合速率分别比WW降低56.7%、26.6%、18.7%和9.5%.干旱胁迫降低了甘薯垄间的日蒸发量、蒸腾速率、耗水量和日耗水量,降低了土壤水利用效率而提高了灌溉水利用效率.干旱胁迫通过降低叶面积系数和光合速率,减少了生物产量及其向块根的分配,进而导致薯干产量显著降低.干旱胁迫时间越早、持续时间越长,对叶面积系数和光合速率,以及生物产量和收获指数的不利影响越大、导致减产幅度越大,水分利用效率越低.在有限的灌水条件下,甘薯生产中应尽可能减少前期干旱.     

Effects of autophagy on acid-sensing ion channel 1a-mediated apoptosis in rat articular chondrocytes

The incidence of spinal cord injuries (SCI) is high every year. As the spinal cord is the highway that allows for the brain to control the rest of the body, spinal cord injuries greatly impact the quality of life of the patients. The SCI include the primary response consisting of the initial accident-induced damage and the secondary response that is characterized by damage due to inflammation and biological responses. Astrocytes are the first to act at the site of the injury, forming a glial scar and attracting immune cells. The immune system plays a role in cleaning out the debris caused by the injury, as well as preventing neurons to grow and heal. The secondary injury caused by the inflammatory response is the major target to combat SCI. This article critically reviews the key players in the inflammatory SCI response and potential therapies, specifically targeting astrocytes, neutrophils, and macrophages. These cells are both beneficial and detrimental following SCI, depending on the released molecules and the types of cells infiltrated to the site of injury. Indeed, depending on the subtype of macrophages, M1 or M2, beneficial or detrimental response could be incited. Therapeutic strategies to regulate and manipulate the immune cells via increasing or decreasing their recruitment to the site of injury could be developed together with upregulating and downregulating the release of certain chemicals from the infiltrated cells.     

家蚕雄dsx~(M3)异位表达对下游靶基因表达的影响

家蚕双性基因dsx通过雌雄特异选择性剪接并表达影响体细胞性别发育。在对Bmdsx的表达情况进行深入分析时发现了一个新的雄特异剪接体dsx~(M3),但其功能未知。为揭示Bmdsx~(M3)的生物学功能,首次通过转基因在雌蚕中异位表达Bmdsx~(M3),成功获得两个转基因品系T1和T2。在T1和T2中虽未出现发育异常和性反转事件,但在转基因雌性个体中,SP1和Vg的表达量都出现明显下调,而PBP的表达量也出现明显上调,表明新剪接体Bmdsx~(M3)能够调控Bmdsx下游靶基因的表达,暗示Bmdsx~(M3)同样参与家蚕体细胞性别发育。     

核糖糖基化BSA单体对SH-SY5Y细胞的毒性明显

研究显示,蛋白质异常修饰形成的寡聚体,与其多聚体、淀粉样纤维相比,具有更强的细胞毒性．这一发现被认为是蛋白质错误折叠和聚集研究领域中的重要进展．蛋白质的异常修饰如还原糖的非酶糖基化,是糖尿病最基本的病理特征．2型糖尿病患者尿液中的核糖浓度显著升高,表明糖尿病不仅与葡萄糖代谢紊乱相关,同时也与核糖代谢失调相关．以牛血清白蛋白(BSA)为研究对象,通过荧光分光光度计检测、原子力显微镜、透射电子显微镜观察以及分子排阻色谱分离,观察到核糖糖基化能够诱导BSA聚集,从单体、寡聚体逐渐形成多聚体．通过CCK-8 Kit、乳酸脱氢酶细胞活性检测、TUNEL染色、caspase-3活性检测以及流式细胞检测等方法,发现核糖糖基化的BSA单体对SH-SY5Y细胞(人神经母细胞瘤细胞系)具有明显的毒性,与此同时,糖化寡聚体和多聚体没有表现出显著的毒性．进一步研究发现,核糖糖基化的BSA单体通过与AGEs的受体RAGE相互作用,激活细胞内的MAPK通路,从而导致细胞凋亡．