A cyp19a1b‐gfp (aromatase B) transgenic zebrafish line that expresses GFP in radial glial cells

Aromatase is an enzyme that catalyzes the synthesis of estrogen in gonads and brain. Teleost fish express aromatase (AroB) strongly in the brain facilitating its detailed examination. To understand the function of AroB in the brain, we generated transgenic zebrafish that expresses green fluorescent protein (GFP) driven by the brain aromatase cyp19a1b promoter. GFP was found in the radial glial cells of transgenic larvae and adult fish that overlap with AroB immunoreactivity in the correct temporal and spatial pattern. GFP was also coexpressed with radial cell marker BLBP, but was not in neurons. In addition, GFP expression in the radial glial cells was stimulated by estrogen, same as endogenous AroB expression. Thus, this transgenic line faithfully mimics the regulation of AroB expression in radial glial cells. It provides a powerful tool to further characterize progenitor radial cells in adult and developing fish and to evaluate estrogenic activities of xenoestrogens and phytoestrogens. genesis 47:67–73, 2009. © 2008 Wiley‐Liss, Inc.     

Mechanisms for the apoptosis of small cell lung cancer cells induced by anti-GD2 monoclonal antibodies: roles of anoikis

Anti-GD2 ganglioside antibodies could be a promising, novel therapeutic approach to the eradication of human small cell lung cancers, as anti-GD2 monoclonal antibodies (mAbs) induced apoptosis of small cell lung cancer cells in culture. In this study, we analyzed the mechanisms for the apoptosis of these cells by anti-GD2 mAbs and elucidated the mechanisms by which apoptosis signals were transduced via reduction in the phosphorylation levels of focal adhesion kinase (FAK) and the activation of a MAPK family member, p38, upon the antibody binding. Knock down of FAK resulted in apoptosis and p38 activation. The inhibition of p38 activity blocked antibody-induced apoptosis, indicating that p38 is involved in this process. Immunoprecipitation-immunoblotting analysis of immune precipitates with anti-FAK or anti-integrin antibodies using an anti-GD2 mAb revealed that GD2 could be precipitated with integrin and/or FAK. These results suggested that GD2, integrin, and FAK form a huge molecular complex across the plasma membrane. Taken together with the fact that GD2+ cells showed marked detachment from the plate during apoptosis, GD2+ small cell lung cancer cells seemed to undergo anoikis through the conformational changes of integrin molecules and subsequent FAK dephosphorylation.     

水产废弃物胶原蛋白的提取

以水产废弃物为原料,利用酶法提取有价值的胶原蛋白。以胃蛋白酶为胶原蛋白提取用酶,鱼鳍和鱼鳞作为提取的原料,提取工艺为原料粉碎、脱钙、提取、纯化。鱼鳞和鱼鳍的胶原蛋白提取率分别达8．1％和6．6％。该方法对提高水产废弃物的综合利用具有重要的意义。     

氯喹对戊四氮致痫大鼠皮质和海马白细胞介素1β及肿瘤坏死因子α表达的影响

目的观察氯喹对戊四氮致痫大鼠皮质和海马白细胞介素1β(IL-1β)及肿瘤坏死因子α(TNF-α)表达的影响,探讨其在癫痫发生发展过程中的作用.方法 48只健康雄性SD大鼠随机分为对照组(12只)、戊四氮(PTZ)致痫组(18只,60mg/kg,i.p.)和氯喹干预组(18只,氯喹0.61mg/kg,i.c.v.,2h后注射PTZ).每组确定6个时间点:1h、2h、4h、8h、12h和24h.观察大鼠行为表现,记录脑电改变,用免疫组化检测皮质和海马IL-1β和TNF-α表达的变化.结果对照组无痫样发作和痫样放电,戊四氮致痫组痫样发作重(Ⅲ-Ⅴ级),氯喹干预组轻(Ⅰ-Ⅲ级)(P<0.05);脑电记录显示戊四氮致痫组呈频发高幅的痫样波,氯喹干预组痫样波幅低且缓;LI-1β和TNF-α在戊四氮致痫组皮质和海马表达强,与对照组比较差异有显著性(P<0.05),氯喹干预组与对照组比较差异无显著性(P>0.05).结论氯喹可能通过对IL-1β和TNF-α表达的抑制减轻戊四氮致痫大鼠的痫样放电和痫样发作程度.这些结果提示,氯喹在防治癫痫方面可能是理想的抗痫剂.     

Five New Guaiane Sesquiterpenes from the Endophytic Fungus Xylaria sp. YM 311647 of Azadirachta indica

Five new guaiane sesquiterpenes, 1 – 5 , were isolated from the culture broth of the endophytic fungus Xylaria sp. YM 311647, isolated from Azadirachta indica A. Juss . The structures of these compounds were elucidated on the basis of spectroscopic analyses, and their inhibitory activities against five pathogenic fungi were evaluated. All guaiane sesquiterpenes showed moderate or weak antifungal activities in a broth microdilution assay.     

LC-MS/MS determination of helicid in human plasma and its application in pharmacokinetic studies

Helicid is a traditional Chinese medicine used to treat headache and insomnia with definite effects. To facilitate pharmacokinetic studies of helicid in man, a sensitive and specific LC-MS/MS method for the quantitative detection of helicid in human plasma was developed and validated. The method involved the addition of bergeninum as the internal standard (IS), protein precipitation, HPLC separation, and quantification by MS/MS system using negative electrospray ionization in the multiple reaction monitoring mode (MRM). The precursor→product ion transitions were monitored at m/z 282.8→120.9 for helicid and m/z 326.9→192.2 for the IS, respectively. The lower limit of quantification (LLOQ) was 0.2 μg/L. The calibration curves for helicid was linear over a concentration range of 0.2-20 μg/L. The intra- and inter-batch analyses of QC samples at 0.4, 2, 20 μg/L indicated good precision (%R.S.D. between 2.69 and 5.47%) and accuracy (between 96.15 and 105.05%). The helicid was stable in human plasma stored at room temperature for at least 24h, 4°C for at least 24h, -20°C for at least 1 month, and for routine three freeze-thaw cycles. This accurate and specific assay provides a useful method for evaluating the pharmacokinetic profile of helicid in humans.     

Progesterone and DNA damage encourage uterine cell proliferation and decidualization through up-regulating ribonucleotide reductase 2 expression during early pregnancy in mice

Embryo implantation into the maternal uterus is a crucial step for the successful establishment of mammalian pregnancy. Following the attachment of embryo to the uterine luminal epithelium, uterine stromal cells undergo steroid hormone-dependent decidualization, which is characterized by stromal cell proliferation and differentiation. The mechanisms underlying steroid hormone-induced stromal cell proliferation and differentiation during decidualization are still poorly understood. Ribonucleotide reductase, consisting of two subunits (RRM1 and RRM2), is a rate-limiting enzyme in deoxynucleotide production for DNA synthesis and plays an important role in cell proliferation and tumorgenicity. Based on our microarray analysis, Rrm2 expression was significantly higher at implantation sites compared with interimplantation sites in mouse uterus. However, the expression, regulation, and function of RRM2 in mouse uterus during embryo implantation and decidualization are still unknown. Here we show that although both RRM1 and RRM2 expression are markedly induced in mouse uterine stromal cells undergoing decidualization, only RRM2 is regulated by progesterone, a key regulator of decidualization. Further studies showed that the induction of progesterone on RRM2 expression in stromal cells is mediated by the AKT/c-MYC pathway. RRM2 can also be induced by replication stress and DNA damage during decidualization through the ATR/ATM-CHK1-E2F1 pathway. The weight of implantation sites and deciduoma was effectively reduced by specific inhibitors for RRM2. The expression of decidual/trophoblast prolactin-related protein (Dtprp), a reliable marker for decidualization in mice, was significantly reduced in deciduoma and steroid-induced decidual cells after HU treatment. Therefore, RRM2 may be an important effector of progesterone signaling to induce cell proliferation and decidualization in mouse uterus.     

Faster STORM using compressed sensing

In super-resolution microscopy methods based on single-molecule switching, the rate of accumulating single-molecule activation events often limits the time resolution. Here we developed a sparse-signal recovery technique using compressed sensing to analyze images with highly overlapping fluorescent spots. This method allows an activated fluorophore density an order of magnitude higher than what conventional single-molecule fitting methods can handle. Using this method, we demonstrated imaging microtubule dynamics in living cells with a time resolution of 3 s.     

哺乳动物季节性繁殖的内源年生物钟及光敏神经环路研究进展

生活在温带和寒带的哺乳动物在长期的进化过程中形成了季节性繁殖的生活史特征。哺乳动物的繁殖功能主要受到下丘脑-垂体-性腺轴(hypothalamic-pituitary-gonadal axis,HPGA)的调控。视交叉上核(suprachiasmatic nucleus,SCN)能够自发振荡并响应光周期信号的变化,引发褪黑素分泌的改变,并介导下游通路中下丘脑甲状腺激素、Kisspeptin和RF酰胺相关肽(RF amide-related peptide,RFRP)的节律性表达变化,从而调控哺乳动物的季节性繁殖。本文综述了哺乳动物季节性繁殖的内源年生物钟调控,并强调了光敏通路中包括甲状腺激素、Kisspeptin和RFRP在季节性繁殖调控中的重要作用。