“T” is for “transgender”: an analysis of children’s picture books featuring transgender protagonists and narrators

ABSTRACT

This project examined 55 picture books featuring transgender, gender expansive or genderqueer protagonists or narrators published between 2008 and 2018. The purpose of the study was to determine how this genre of children’s literature supports and challenges four gender assumptions: the gender binary, gender essentialism, sex/gender congruency and gender stability. Additionally, this critical analysis explored misgendering within this genre and themes of social rejection and acceptance. Protagonists and narrators were permitted a degree of gender nonconformity, however, the majority of picture books missed opportunities for a more complete exploration of gender possibilities.     

大花胡麻草环烯醚萜合酶基因的克隆与表达分析

目的克隆大花胡麻草环烯醚萜合酶基因(CgIS),并进行表达分析。方法以大花胡麻草根、茎、叶转录组中唯一的CgIS基因序列为基础,采用RT-PCR技术从大花胡麻草幼叶克隆CgIS基因,并进行组织特异性表达分析。结果大花胡麻草CgIS基因(GenBank登录号MH794270)全长1 185 bp,编码394个氨基酸;CgIS蛋白相对相对分子质量44 670,理论pI为6.17;该蛋白属于孕酮5β-还原酶(P5β-R)家族成员,可能定位于细胞质;该蛋白无信号肽,为亲水稳定蛋白,主要由α-螺旋(40.61%)和无规则卷曲(46.70%)构成;该蛋白具有SDR(短链脱氢酶/还原酶)和P5βR蛋白保守结构域;CgIS蛋白与芝麻SiIS蛋白亲缘关系最近;CgIS基因主要在叶中表达。结论克隆了CgIS基因,并对其进行表达分析,为进一步研究该基因的功能和环烯醚萜类的生物合成途径奠定基础。     

miR-513a-5p慢病毒载体的构建及其对人骨肉瘤细胞放疗敏感性的影响

目的:构建miR-513a-5p慢病毒过表达载体，转染人骨肉瘤细胞株，观察miR-513a-5p对人骨肉瘤细胞放疗敏感性的影响。方法:PCR法扩增人miR-513a-5p基因，克隆入pLentis-CMV-GFP-MCS-PGK-PURO载体获得重组质粒pLentis-miR513a，双酶切鉴定并测序后将正确的重组质粒和对照质粒转染293FT细胞制备慢病毒，分别转染骨肉瘤HOS和U2OS细胞，qRT-PCR法及荧光显微镜鉴定转染结果。克隆形成实验、MTT法检测miR-513a-5p高表达HOS和U2OS细胞在X射线照射下细胞存活情况。结果:双酶切及测序结果确定成功构建miR-513a-5p慢病毒载体pLentis-miR513a。qRT-PCR结果提示，转染骨肉瘤细胞株后miR-513a-5p表达显著升高。克隆形成实验结果显示miR-513a-5p高表达后骨肉瘤细胞在X射线照射下细胞增殖减慢。MTT结果提示miR-513a-5p高表达骨肉瘤细胞经X射线照射后细胞存活减少。结论:成功构建了miR-513a-5p慢病毒载体，建立了高效稳定表达miR-513a-5p的骨肉瘤细胞株，高表达miR-513a-5p能显著增加X射线照射后骨肉瘤细胞的放疗敏感性。     

人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

目的：克隆人肠道病毒71型（EV71）及柯萨奇病毒A组16型（CA16）的VP1蛋白编码基因，构建相应真核表达质粒在体外进行重组表达，检测其细胞内定位，为EV71及CA16的疫苗研究提供基础。方法：通过PCR扩增分别获取EV71及CA16的VP1编码序列，插入pcFlag载体，分别构建EV71及CA16 VP1与FLAG标签融合的真核表达质粒；瞬时转染人胚肾293T细胞及横纹肌肉瘤RD细胞，Western blot法检测融合蛋白的表达，免疫荧光检测融合蛋白的细胞内定位情况。结果：测序表明两种来源VP1蛋白的重组表达质粒构建正确；分别转染两种不同的细胞后，均可通过Western blot检测到相应蛋白表达；免疫荧光检测显示两种毒株的VP1均表达在细胞质中。结论：外源融合表达EV71或CA16 VP1蛋白的真核表达质粒构建成功；所构建质粒分别转染细胞后，均可在细胞质内检测到VP1融合蛋白的表达。     

Recent advances in preclinical in vitro approaches towards quantitative prediction of hepatic clearance and drug-drug interactions involving organic anion transporting polypeptide (OATP) 1B transporters

Hepatic uptake mediated by organic anion transporting polypeptide (OATP) 1B1 and 1B3 can serve as a major elimination pathway for various anionic drugs and as a site of drug-drug interactions (DDIs). This article provides an overview of the in vitro approaches used to predict human hepatic clearance (CL_h) and the risk of DDIs involving OATP1Bs. On the basis of the so-called extended clearance concept, in vitro–in vivo extrapolation methods using human hepatocytes as in vitro systems have been used to predict the CL_h involving OATP1B-mediated hepatic uptake. CL_h can be quantitatively predicted using human donor lots possessing adequate OATP1B activities. The contribution of OATP1Bs to hepatic uptake can be estimated by the relative activity factor, the relative expression factor, or selective inhibitor approaches, which offer generally consistent outcomes. In OATP1B1 inhibition assays, substantial substrate dependency was observed. The time-dependent inhibition of OATP1B1 was also noted and may be a mechanism underlying the in vitro–in vivo differences in the inhibition constant of cyclosporine A. Although it is still challenging to quantitatively predict CL_h and DDIs involving OATP1Bs from only preclinical data, understanding the utility and limitation of the current in vitro methods will pave the way for better prediction.     

抑郁症大鼠模型海马区及前额叶组织BDNF因子、Trkb蛋白表达的实验研究

目的:观察抑郁症大鼠模型脑组织不同部位神经营养因子水平的变化及抗抑郁药物使用后改变。方法:将成年SD大鼠30只分为正常对照组、模型对照组、药物干预组,每组10只,雌雄对半;正常组正常养殖,模型组采用慢性不可预知应激结合孤养方式制备抑郁模型大鼠4周。药物干预组采用模型组造模过程4周后给予药物(氟西汀)给入,自实验开始之后每周观察大鼠体重、糖水消耗、旷场实验指标的变化,最后采用荧光定量PCR法观察大鼠脑部BDNF因子及Trkb的表达。结果:与正常对照组大鼠相比,模型组大鼠体重增加缓慢、糖水消耗减少、旷场试验各项指标较正常组间差异具有统计学意义;荧光定量PCR结果显示:模型组较对照组各脑区BDNF及Trkb受体表达减少,药物干预组脑区BDNF及Trkb受体表达显著高于模型对照组。结论:脑部BDNF因子及Trkb的表达在抑郁症发生与治疗中有所改变。