正交试验结合熵权TOPSIS优选麻黄-桂枝药对水提工艺

目的 优选麻黄-桂枝药对的水提工艺。方法 建立麻黄-桂枝药对超高效液相色谱法指纹图谱,采用Ｌ₉（3⁴）正交试验设计,以煎煮次数、加水量、煎煮时间为考察因素,以指纹图谱及醇溶性浸出物的含量为评价指标,结合熵权逼近理想解排序法（TOPSIS）对正交水提工艺进行综合评价。结果 优选麻黄-桂枝药对最佳水提工艺为加10倍量水煎煮3次,每次煎煮30 min。结论 优选的麻黄-桂枝药对水提工艺科学合理、稳定可行,可为麻黄-桂枝药对的工业生产提供参考。     

Strategic expression of ion transport peptide gene products in central and peripheral neurons of insects

Structurally related ion transport peptides (ITP) and crustacean hyperglycemic hormones (CHH) are increasingly implicated in diverse metabolic and developmental functions in arthropods. We identified a conserved ITP gene encoding two peptides by alternative splicing in Manduca sexta, Bombyx mori, and Aedes aegypti: A C-terminally amidated ITP and a C-terminally unblocked ITP-like peptide (ITPL), which share common N-terminal sequences but have divergent C-termini. In the moth M. sexta, these peptides are expressed in two, regionally distinct neuronal populations in the central and peripheral nervous systems (CNS, PNS). MasITP expression is confined to the brain in five pairs of lateral neurosecretory cells (type Ia(2)) projecting ipsilateral axons into the retrocerebral complex and three to five pairs of adjacent small neurons that arborize extensively within the brain. Expression of MasITPL is comparatively weak in the brain but strong in the ventral ganglia and the PNS, where MasITP is absent. MasITPL occurs in bilaterally paired neurons of all thoracic and abdominal ganglia. In the PNS, MasITPL is coexpressed with crustacean cardioactive peptide in type II link nerve neurons (L1) of abdominal segments 2-7, which project axons into neurohemal transverse nerves. During metamorphosis, additional expression of MasITPL is observed in two pairs of small lateral neurons in the brain and one pair of ventromedial neurons in each of AG2-6. A similar pattern of differential ITP and ITPL expression was observed in the CNS and PNS of B. mori and Schistocerca americana. These distinctive cellular expression patterns suggest that ITP and ITPL have evolved specialized physiological functions in arthropods.     

Identification and expression of the achaete-scute complex in the silkworm, Bombyx mori

Recently, the study of achaete-scute ( AS-C ) homologues has contributed enormously to understanding of gene duplication and function evolution, particularly in Diptera. We identified four AS-C homologue genes in the silkworm, Bombyx mori , referred to as BmASH , BmASH2 , BmASH3 , and Bmase . The complex displayed tandem array structure in the genome. Analysis of spatial expression profiles showed that they all were expressed in obviously higher levels in wing disc than in other tissues, suggesting that they might play important roles in the development of the wing. Furthermore, we found that their expression profiles in the wing discs were mostly correlated with the development of the scales, especially the BmASH gene. RNA interference results further indicated that BmASH was necessary for scale formation in silkworm wing.     

杆状病毒多基因表达系统介导的轮状病毒样颗粒表达与组装

目的 构建能同时表达轮状病毒vp2、vp6、vp7基因的重组家蚕杆状病毒,并在BmN细胞中进行初步表达.方法 利用MA104细胞繁殖人A组轮状病毒,RT-PCR扩增病毒结构蛋白vp2、vp6、vp7基因,连接至转移载体pFBDM和pUCDM中,同时引入含IE1启动子的绿色荧光蛋白基因利于检测感染和表达量,再分别通过Tn7及Cre-LoxP重组构建重组病毒.为方便检测基因的表达,在VP7的C-末端添加6-组氨酸(6-His)标签,可通过ELISA等方法快速检测目的 基因表达.结果 扩增获得了轮状病毒三个结构蛋白基因,构建了能高效组装轮状病毒样颗粒的重组表达载体,ELISA检测VP7得到了表达,在BmN细胞中观察到病毒样颗粒.结论 成功构建杆状病毒多基因的表达系统,为生产多哑基蛋白复合物,研究大分子结构提供借鉴.     

液相微萃取对中药中低丰度有效成分桂皮酸及其衍生物的浓缩富集及色谱分析

目的利用液相微萃取/后萃取-高效液相色谱法(HPLC)对中药中低丰度有效成分桂皮酸及衍生物进行定性鉴别及含量测定,探讨欲萃物浓度、供相体积与富集倍数之间的关系。方法正庚醇为萃取溶剂,供相为2.0×10-4mol/LHCl,接收相为0.008mol/LNaOH,1800r/min转速下,萃取60min后进行HPLC分析。结果桂皮酸、对羟基桂皮酸、对甲氧基桂皮酸的线性范围可达6个数量级,检测限分别为0.2,0.08,0.2pg/ml。精密度:日内RSD<10.6%,日间RSD<11.9%,回收率为94.7%～101.3%,92.1%～105.9%。结论该方法操作简单,分析物的富集倍数达到近100倍,对常规前处理方法难以分析的低丰度微量、痕量成分的分析具有重要的意义。     

Bombyx mori Ras proteins BmRas1, BmRas2 and BmRas3 are neither farnesylated nor palmitoylated but are geranylgeranylated

The lipid modifications which occur on Bombyx mori Ras proteins BmRas1, BmRas2 and BmRas3 were studied by metabolic labelling in an insect cell‐free protein synthesis system and in a baculovirus expression system, using specific inhibitors of protein prenylation and protein palmitoylation. In addition, the subcellular localization of BmRas proteins was examined using EGFP fusion proteins of constitutively active forms of BmRas proteins transiently expressed in Sf9 cells. As a result, it was revealed that the three B. mori Ras proteins BmRas1, BmRas2 and BmRas3 are neither farnesylated nor palmitoylated but are geranylgeranylated for localization to the plasma membrane of insect cells. Thus, the mechanism of membrane binding of insect Ras proteins is quite different from that reported for mammalian Ras proteins.     

桑枝多糖对糖尿病肾病大鼠肾脏组织抗氧化作用的影响

目的：研究桑枝多糖对链脲佐菌素（STZ）诱导糖尿病大鼠的血糖和抗氧化作用的影响。方法：STZ（120 mg·kg^-1）腹腔注射诱导糖尿病大鼠模型,并随机分为5个组（n=10）：模型对照组,二甲双胍对照组（320 mg·kg^-1）,桑枝多糖低、中、高剂量组（0.3,0.6,1.2 g·kg^-1）,另设10只大鼠作为空白对照组。大鼠灌胃给药60 d,并测定大鼠空腹血糖（FBG）。终点法测24 h尿微量白蛋白含量。给药60 d后处死大鼠,对大鼠肾脏HE染,光镜下观察肾脏病理学的变化;试剂盒测定大鼠肾脏中锰超氧化物歧化酶（Mn-SOD）、谷胱甘肽过氧化物酶（GSH-Px）的活性和丙二醛（MDA）含量;比色法检测肾组织中线粒体呼吸链复合物Ⅰ、Ⅲ活性。结果：与模型组比较,糖尿病大鼠经桑枝多糖治疗后血糖有所下降并显著降低尿微量白蛋白含量（P<0.05）。模型组大鼠肾脏中Mn-SOD、GSH-Px活性以及肾组织线粒体呼吸链复合物Ⅰ、Ⅲ活性显著降低,而MDA含量则升高。桑枝多糖给药后Mn-SOD、GSH-Px、线粒体呼吸链复合物Ⅰ、Ⅲ活性有所升高,MDA含量下降（P<0.05）。结论：桑枝多糖能显著降低糖尿病大鼠的血糖,其作用机制可能与提高糖尿病大鼠的抗氧化作用有关。     

基因工程人丁酰胆碱酯酶的纯化及其在体外对琥珀酰胆碱的解毒作用(英文)

目的　开发丰富又安全的人丁酰胆碱酯酶来源 ,并检测其解毒效率。方法　家蚕血淋巴液中重组人丁酰胆碱酯酶 (rhBChE)的纯化采用批量硫酸铵分段分离和普鲁卡因酰胺 脂糖凝胶 4B亲和层析技术。结果　聚丙烯酰胺凝胶电泳纯的rhBChE从血淋巴液中的总收率为 6 6 % ,酶比活性达 715mmol·min- 1·g- 1蛋白。rhBChE在 - 2 0℃含 2 0 %甘油及0 .0 2 %NaN3 的 10mmol·L- 1磷酸盐缓冲液 (pH 7.4 )中存放 1年 ,酶活性无明显变化。 18只小鼠ip事先与rhBChE孵温的 1.5LD的琥珀酰胆碱 ,不出现任何中毒症状和体征 ,全部存活 ;而琥珀酰胆碱对照组 ,18只小鼠均剧烈抽搐并在 2～ 5min内窒息死亡。结论　实验结果说明 ,从感染家蚕幼虫血淋巴液中纯化rhBChE的方法可靠 ,基因工程rhBChE在琥珀酰胆碱中毒危象的治疗中有潜在应用前景。