Reactive oxygen and nitrogen species during meiotic resumption from diplotene arrest in mammalian oocytes

Mammalian ovary is metabolically active organ and generates by‐products such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) on an extraordinary scale. Both follicular somatic cells as well as oocyte generate ROS and RNS synchronously and their effects are neutralized by intricate array of antioxidants. ROS such as hydrogen peroxide (H₂O₂) and RNS such as nitric oxide (NO) act as signaling molecules and modulate various aspects of oocyte physiology including meiotic cell cycle arrest and resumption. Generation of intraoocyte H₂O₂ can induce meiotic resumption from diplotene arrest probably by the activation of adenosine monophosphate (AMP)‐activated protein kinase A (PRKA)—or Ca²⁺‐mediated pathway. However, reduced intraoocyte NO level may inactivate guanylyl cyclase‐mediated pathway that results in the reduced production of cyclic 3′,5′‐guanosine monophosphate (cGMP). The reduced level of cGMP results in the activation of cyclic 3′,5′‐adenosine monophosphate (cAMP)‐phosphodiesterase 3A (PDE3A), which hydrolyses cAMP. The reduced intraoocyte cAMP results in the activation of maturation promoting factor (MPF) that finally induces meiotic resumption. Thus, a transient increase of intraoocyte H₂O₂ level and decrease of NO level may signal meiotic resumption from diplotene arrest in mammalian oocytes. J. Cell. Biochem. 111: 521–528, 2010. © 2010 Wiley‐Liss, Inc.     

我国丹顶鹤栖息地选择研究进展

丹顶鹤(Grus japonensis)是我国Ⅰ级重点保护野生动物,被世界自然保护联盟列为濒危物种。本研究通过搜集和整理2000至2017年丹顶鹤栖息地选择和生境适宜性相关文献,分析了丹顶鹤在繁殖期、迁徙期、越冬期的栖息地选择,发现丹顶鹤在这三个时期的最偏好生境均为芦苇沼泽或芦苇滩,回避人为干扰较为严重的居民点、道路、盐田等生境。通过梳理丹顶鹤繁殖地、中途停歇地、越冬地的栖息地变化及其原因,发现丹顶鹤栖息地变化是自然因素和人为因素共同驱动的结果,以人为因素为主。本研究希望为科学评价栖息地变化对丹顶鹤野生种群的影响、针对性地开展栖息地保护管理工作、促进丹顶鹤野生种群健康可持续发展提供依据。     

人脐带间充质干细胞重复静脉注射动物毒性试验

摘要 目的：评价多次尾静脉注射脐带间充质干细胞（hUC-MSCs）对小鼠的体内毒性作用。方法：48只健康ICR小鼠,按性别和体重随机分为4组（即对照组、低剂量组、中剂量组和高剂量组）。小鼠通过微静脉注射不同剂量hUC-MSCs悬浮液,间隔3天给药1次,共给药4次。记录小鼠摄食量、体重、体温,给药结束后恢复两周后牺牲动物作大体解剖,检查各个器官器质性病变;利用流式细胞仪分别检测CD3、CD4、CD8阳性细胞亚群数量;ELISA试剂盒检测血清IgM、IgG、C3、C4指标;对肺脏、脾脏、肾脏行组织病理学检查。结果：实验组与对照组相比较,注射不同剂量干细胞后一般观察、体重、体温、摄食量、IgM以及C3在给药期和恢复期均未发生显著变化。在恢复期,注射中、高剂量hUC-MSCs组血清IgG和C4水平略有降低,但未达到显著水平P<0.05;CD4阳性T细胞集群数量以及CD4/CD8系数在hUC-MSCs中、高剂量组显著上升（P<0.05）。大体剖检,除脾脏相比溶媒对照组略显增大外其它各器官均未发现肉眼可见明显异常;称重后发现hUC-MSCs高剂量组脾重量与溶媒对照组相比显著升高（P<0.05）。脾脏、肺脏、肾脏病理学检测未见明显异常。结论：健康ICR小鼠尾静脉注射临床剂量hUC-MSCs（1×10⁶ cells/kg）可能调动动物免疫反应,此外,未观察到hUC-MSCs对小鼠有明显毒副作用。     

Bile acids,FGF15/19 and liver regeneration: From mechanisms to clinical applications

The liver has an extraordinary regenerative capacity rapidly triggered upon injury or resection. This response is intrinsically adjusted in its initiation and termination, a property termed the “hepatostat”. Several molecules have been involved in liver regeneration, and among them bile acids may play a central role. Intrahepatic levels of bile acids rapidly increase after resection. Through the activation of farnesoid X receptor (FXR), bile acids regulate their hepatic metabolism and also promote hepatocellular proliferation. FXR is also expressed in enterocytes, where bile acids stimulate the expression of fibroblast growth factor 15/19 (FGF15/19), which is released to the portal blood. Through the activation of FGFR4 on hepatocytes FGF15/19 regulates bile acids synthesis and finely tunes liver regeneration as part of the “hepatostat”. Here we review the experimental evidences supporting the relevance of the FXR-FGF15/19-FGFR4 axis in liver regeneration and discuss potential therapeutic applications of FGF15/19 in the prevention of liver failure. This article is part of a Special Issue entitled: Cholangiocytes in Health and Disease edited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen.     

土壤Cd胁迫对三七生长和根系DNA损伤及抗氧化酶活性的影响

采用盆栽实验法,研究了土壤中添加0.0(对照)、0.1、0.3、0.6、1.0、3.0、6.0、10.0和30.0 mg·kg-1Cd对三七〔Panax notoginseng(Burk.)F.H.Chen〕生长和抗氧化酶活性的影响,并采用彗星实验对Cd胁迫条件下三七根尖细胞的DNA损伤进行了分析。结果显示:随土壤Cd添加量增加,三七的成活率、株高、单株复叶数和单株叶面积以及单株根、茎和叶片的干质量及鲜质量总体上呈先升高后降低的趋势;其中,各处理组三七植株的成活率和单株复叶数均高于对照,而株高和单株叶面积则在Cd添加量较低的条件下高于对照、在Cd添加量较高的条件下低于对照;单株根、茎和叶的鲜质量及干质量在Cd添加量30 mg·kg-1条件下均小于对照但差异不显著。随Cd添加量的提高,三七根系中SOD和CAT活性呈先上升后下降、再上升再下降的变化趋势,而POD活性总体上逐渐升高;其中,Cd添加量0.6、1.0、3.0和30.0 mg·kg-1处理组的SOD活性极显著或显著低于对照,而各处理组的POD和CAT活性总体上均高于对照且在Cd添加量1.0~30.0 mg·kg-1条件下与对照有极显著差异。彗星实验结果表明:各处理组三七根尖细胞的彗尾长、尾部DNA相对含量和Olive尾矩均有差异,其中,在Cd添加量较高的条件下各指标均高于对照但差异均不显著。研究结果显示:在较低水平的土壤Cd胁迫条件下,三七的生长、抗氧化酶活性和根系DNA均没有受到明显伤害,而较高水平的Cd胁迫则对其生长和抗氧化酶活性有抑制作用,且根尖细胞DNA损伤也较严重。     

TRAF6 promotes TGFβ-induced invasion and cell-cycle regulation via Lys63-linked polyubiquitination of Lys178 in TGFβ type I receptor

Transforming growth factor β (TGFβ) can act either as a tumor promoter or a tumor suppressor in a context-dependent manner. High levels of TGFβ are found in prostate cancer tissues and correlate with poor patient prognosis. We recently identified a novel TGFβ-regulated signaling cascade in which TGFβ type I receptor (TβRI) is activated by the E3 ligase TNF-receptor-associated factor 6 (TRAF6) via the Lys63-linked polyubiquitination of TβRI. TRAF6 also contributes to activation of TNF-α-converting enzyme and presenilin-1, resulting in the proteolytic cleavage of TβRI and releasing the intracellular domain of TβRI, which is translocated to the nucleus to promote tumor invasiveness. In this report, we provide evidence that Lys178 of TβRI is polyubiquitinated by TRAF6. Moreover, our data suggest that TRAF6-mediated Lys63-linked ubiquitination of the TβRI intracellular domain is a prerequisite for TGFβ regulation of mRNA for cyclin D1 (CCND1), expression, as well as for the regulation of other genes controlling the cell cycle, differentiation, and invasiveness of prostate cancer cells.     

CCCTC-结合因子的表达水平影响其在细胞内的分布

目的:研究CCCTC-结合因子(CTCF)的表达水平与其在HeLa和HepG2细胞内分布的关系。方法:利用小干扰RNA敲降CTCF的表达;利用免疫荧光染色检测CTCF在细胞内的分布。结果:在HeLa和HepG2细胞中下调CTCF的表达,检测到CTCF在细胞核内的分布比例减少,而在细胞质内的分布比例相应增加。结论:CTCF的表达水平会影响其在细胞内的分布。     

Protein–protein interactions between SWCNT/chitosan/EGF and EGF receptor: a model of drug delivery system

Epidermal growth factor (EGF) was used as the targeting ligand to enhance the specificity of a cancer drug delivery system (DDS) via its specific interaction with the EGF receptor (EGFR) that is overexpressed on the surface of some cancer cells. To investigate the intermolecular interaction and binding affinity between the EGF-conjugated DDS and the EGFR, 50 ns molecular dynamics simulations were performed on the complex of tethered EGFR and EGF linked to single-wall carbon nanotube (SWCNT) through a biopolymer chitosan wrapping the tube outer surface (EGFR·EGF-CS-SWCNT-Drug complex), and compared to the EGFR·EGF complex and free EGFR. The binding pattern of the EGF-CS-SWCNT-Drug complex to the EGFR was broadly comparable to that for EGF, but the binding affinity of the EGF-CS-SWCNT-Drug complex was predicted to be somewhat better than that for EGF alone. Additionally, the chitosan chain could prevent undesired interactions of SWCNT at the binding pocket region. Therefore, EGF connected to SWCNT via a chitosan linker is a seemingly good formulation for developing a smart DDS served as part of an alternative cancer therapy.     

大萼香茶菜中的二萜化学成分研究

为研究大萼香茶菜(Isodon macrocalyx )的化学成分,该文采用硅胶、ODS、Sephadex LH-20、反相C₁₈半制备高效液相等色谱方法对大萼香茶菜地上部分进行分离纯化,并利用¹H NMR、¹³C NMR和HR-ESI-MS等波谱数据,以及结合 参考文献,鉴定了这些化合物的结构。结果表明:从大萼香茶菜地上部分分离得到13个二萜,它们分别是19-羟基陶塔酚(1)、macrophynin E(2)、inumakoic acid(3)、inumakiol D(4)、4β-carboxy-19-nortotarol(5)、(-)-lambertic acid(6)、2-oxo-5-fagonene(7)、isodoterniofiln B(8)、长管贝壳杉素E(9)、长管香茶菜素A(10)、牛尾草素H(11)、16S-dihydrolongikaurin A(12)和ent-3S,16S,17-trihydroxy-kauran-2-one(13)。所有得到的二萜均为首次从该植物中分离得到。     

Modular bioreactor for primary human hepatocyte culture: medium flow stimulates expression and activity of detoxification genes

Down-regulation of detoxification genes, notably cytochrome P450 (CYPs), in primary hepatocyte cultures is a long-standing and major concern. We evaluated the influence of medium flow in this model. Hepatocytes isolated from 12 different liver donors were cultured either in a multichamber modular bioreactor (MCmB, flow rate 250-500 μL/min) or under standard/static conditions, and the expression of 32 genes, enzyme activities and biological parameters were measured 7-21 days later. mRNA expression of genes involved in xenobiotic/drug metabolism and transport, including CYP1A1, 1A2, 2B6, 2C9, 3A4 (and activities for some of them), UDP-glucuronosyltransferase (UGT) 1A1, UGT2B4, UGT2B7, glutathione S-transferase (GSTα), and multidrug resistance protein 1 (MDR1) and MRP2, were specifically up-regulated by medium flow as compared with static controls in all cultures tested. In 2-week-old cultures, expression of detoxification genes reached levels close to or higher than those measured in freshly isolated hepatocytes. In contrast, CYP2D6 and most of other tested genes were not affected by medium flow. We conclude that medium flow specifically interferes with, and up-regulates, the activity of xenosensors and/or the expression of detoxification genes in primary human hepatocytes. Down-regulation of detoxification genes in conventional (static) cultures is therefore partly a consequence of the absence of medium circulation.