解毒扶正方对肺癌分子靶向治疗获得性耐药模型小鼠的干预作用探讨

目的：观察解毒扶正方对分子靶向药物获得性耐药非小细胞肺癌小鼠模型的干预作用。方法：选取BALB/C nu/nu小鼠40只,皮下接种人非小细胞肺癌细胞株PC9/GR成瘤构建皮下移植瘤,随机分为5组,即空白对照组、低剂量+阳性组、高剂量+阳性组、高剂量组、阳性对照组,每组各8只。所有动物经口灌胃给药,灌胃剂量：中药（解毒扶正方）低剂量为52 g生药/kg体质量,中药高剂量为104 g生药/kg体质量,阳性药物（吉非替尼）为51.38 mg/kg体质量,空白对照组给予纯化水灌胃,每天1次,连续15 d。观察各组的瘤体体积、肿瘤生长抑制率,观察瘤体组织,计算肿瘤细胞增殖指数,免疫组化法检测蛋白激酶B（AKT）、磷酸化蛋白激酶B（p-AKT）、细胞外信号调节激酶（ERK）、磷酸化细胞外信号调节激酶（p-ERK）及核因子E₂相关因子2（Nrf2）表达水平,计算p-ERK/ERK、p-AKT/AKT值。结果：瘤体体积、肿瘤生长抑制率高剂量组、高剂量+阳性组与空白对照组比较,差异具有统计学意义（P＜0.05）;低剂量+阳性组、高剂量+阳性组、高剂量组肿瘤细胞增殖指数以及肿瘤组织p-ERK/ERK与空白对照组比较,差异具有统计学意义（P＜0.05或P＜0.01）。结论：解毒扶正方可缩小分子靶向药物获得性耐药非小细胞肺癌裸鼠皮下移植瘤模型体积,抑制皮下移植瘤生长,降低肿瘤细胞增殖指数,降低p-ERK/ERK比值,升高Nrf2蛋白表达。     

热带东印度洋春季浮游植物群落结构空间特征分析

基于2013年3月至5月采集的热带东印度洋海域(10.0°S～4.0°N, 83.0°～97.5°E)浮游植物水样样品,分析了其种类组成、优势类群、细胞丰度等群落特征参数,综合比较了水平和垂向上浮游植物种类及丰度的差异性,初步探讨了其成因。结果表明:2013年春季热带东印度洋微型浮游植物共鉴定出306种,主要由硅藻、甲藻、金藻、蓝藻、裸藻和隐藻六大门类组成,其优势类群主要以粒径较小的隐藻、微型甲藻、菱形藻、环沟藻等为主。水平分布上,各水层浮游植物细胞丰度分布趋势相似,但斑块特征明显,其高值区位于88°E断面赤道以南次表层水域(30 m、75 m),局部区域细胞丰度值可达104 cells/L以上;与其毗邻的东南部、东部水域为低值区,并向赤道延伸。垂直剖面上,硅藻和甲藻广泛分布在各取样深度,但分布特征有明显的空间差异和规律,0 m、30 m大部分站位硅藻种类比例在0.2～0.3甚至更低,甲藻在0.7～0.8,随着水层加深(75 m、100 m、150 m、300 m),硅藻种类占比上升到0.5～0.6,甲藻降低到0.4～0.5,无论硅藻还是甲藻种类数75 m层最丰富。30 m和75 m水层细胞丰度明显高于其他水层。甲藻是热带东印度洋微型浮游植物种类和细胞丰度的重要贡献者,低生物量海域表现的更为明显,贡献率大于80%。该研究将极大丰富东印度洋浮游植物群落空间特征基础信息较匮乏的现状,为量化、评估该海域的生物资源提供数据支撑。     

近江牡蛎鳃细胞的原代培养

采用改良的DMEM(HG)培养基,建立了近江牡蛎(Crassostrea hongkongensis)鳃细胞体外培养技术,包括鳃组织块培养法和胰酶消化培养法。结果表明,组织块培养法接种6 h后,细胞开始从组织块中迁出,细胞形态较小,呈圆形、椭圆形或多边形,直径3～6μm。培养至3 d时,细胞在组织块周围形成生长晕。培养至6 d时可进行细胞传代,本次实验细胞已传至第6代。胰酶消化法接种约2 h后,细胞逐渐贴壁,从形态上主要分为两类,一类为小型细胞,形态为圆形、椭圆形或多边形,直径3～6μm,数量多,增殖速度较快;另一类为大型细胞,形态为圆形、椭圆形或多边形,直径10～20μm,部分细胞内部含有颗粒,数量较少,增殖速度较慢。培养至2 d时可进行细胞传代,传代培养物中的优势细胞皆为小型细胞。本次实验细胞已传至第6代。     

久效磷对雄性金鱼分泌细胞超微结构的影响

利用电镜技术观察了久效磷暴露条件下,金鱼中腺垂体促性腺激素细胞、生长激素细胞和促甲状腺激素细胞以及精巢Leydig氏细胞超微结构的变化。结果表明,久效磷对中腺垂体分泌细胞及精巢Leydig氏细胞膜系统的损伤最为严重,主要表现为垂体3种分泌细胞核膜水肿、局部溶解,染色质有不同程度的收缩现象,粗面内质网水肿,部分溶解呈空泡状;精巢Leydig氏细胞核膜局部溶解,染色质轻微凝聚,粗面内质网水肿,局部溶解,线粒体内嵴水肿、局部溶解。久效磷对金鱼中腺垂体促性腺激素细胞、生长激素细胞和促甲状腺激素细胞及精巢Leydig氏细胞结构的损伤是影响内分泌细胞激素分泌功能的原因之一。     

Expression and localization of a novel phosducin-like protein from amphioxus <Emphasis Type="Italic">Branchiostoma belcheri</Emphasis>

A full length amphioxus cDNA, encoding a novel phosducin-like protein (Amphi-PhLP), was identified for the first time from the gut cDNA library of Branchiostoma belcheri. It is comprised of 1 550 bp and an open reading frame (ORF) of 241 amino acids, with a predicted molecular mass of approximately 28 kDa. In situ hybridization histochemistry revealed a tissue-specific expression pattern of Amphi-PhLP with the high levels in the ovary, and at a lower level in the hind gut and testis, hepatic caecum, gill, endostyle, and epipharyngeal groove, while it was absent in the muscle, neural tube and notochord. In the Chinese Hamster Ovary (CHO) cells transfected with the expression plasmid pEGFP-N1/Amphi-PhLP, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that Amphi-PhLP is a cytosolic protein. This work may provide a framework for further understanding of the physiological function of Amphi-PhLP in B. belcheri. Supported by the Pilot Projects of Knowledge Innovation Project of Chinese Academy of Sciences (No. KZCX2-YW-211-03 and MGE2008KG06)     

东海原甲藻线粒体细胞色素b(Cytb)基因的定量检测

建立了检测东海原甲藻线粒体细胞色素b(Cytb)基因mRNA含量的实时荧光定量PCR方法.以甲藻Cytb基因的简并引物扩增得到984 bp长的东海原甲藻Cytb基因片段,经克隆、测序,制备并纯化质粒.以光度法测定质粒浓度并作为标准品.对上述基因片段,利用PRIMER EXPRESS 3.0 设计引物,以梯度稀释的质粒标准品进行定量PCR反应,以SYBR Green I为荧光染料,制作标准曲线,得到基因拷贝数X与Ct值的关系为:Ct=-3.50 lg X+39.35(相关系数r为0.999).通过对不同生长时期的东海原甲藻样品的Cytb基因mRNA含量检测,发现该基因的表达量较稳定,平均值为(45.4±4.7)拷贝/细胞,受生长状态影响不大,可作为实时荧光定量PCR的内参基因.     

Experimental Study on the Interspecific Interactions Between the Two Bloom-Forming Algal Species and the Rotifer Brachionusplicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms （HAB） species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×10^4 cells mL^-1 of P. donghaiense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities （5, 10, 15 and 20 × 10^4 cells mL^-1）, and the increase rate of B. plicatilis population （r） at this algal density was 0.104 ± 0.015 rd^-1. Cell densities of P. donghaiense decreased due to the grazing of B plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.     

Experimental Study on the Interspecific Interactions Between the Two Bloom-Forming Algal Species and the Rotifer Brachionus plicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms(HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×104 cells mL-1 of P. donghaiense,the number of B. plicatilis increa...