球结膜下注射不同浓度消旋山莨菪碱液对雏鸡形觉剥夺近视眼轴的影响

目的研究不同浓度消旋山莨菪碱液对雏鸡形觉剥夺近视眼轴的影响。方法 3日龄雏鸡60只右眼遮盖半透明塑料膜,随机分为6组:给药组4组,不同浓度消旋山莨菪碱液(0.05%组,0.1%组,1%组,2.5%组),隔天行右眼球结膜下注射,每次给药50μL,共4次;对照组为生理盐水组,注射方法同给药组;单纯形觉剥夺组,不做干预。8 d后观察药物对眼轴的影响。结果 2.5%浓度组右眼与左眼眼轴差异显著小于低浓度组及对照组(P<0.05),给药组未发现组织病理毒性。结论消旋山莨菪碱对雏鸡近视眼轴增长有一定抑制作用,无毒副作用,有一定浓度依赖性。     

马传染性贫血病毒基因非编码区LTR嵌合克隆的构建

在已有全长感染性克隆pLGFD3-8和pD70344的基础上,根据马传贫弱毒疫苗致弱过程中不同代次毒株LTR序列的分析,在LTR U3区选取特定的酶切位点对EIAV非编码区LTR基因进行了部分替换.将替换的全基因克隆转染驴胎皮肤细胞(FDD)并以驴白细胞(DL)传代,用逆转录酶活性检测、RT-PCR方法及Real-time RT-PCR验证其感染性.结果发现,其衍生病毒感染上述两种细胞均出现明显的细胞病变;细胞培养上清可检测到RT酶活性和RT-PCR阳性.电镜下可见大量典型的EIAV颗粒.pLGFD9-12嵌合克隆衍生病毒与其父本克隆衍生病毒pLGFD3-8具有相似的复制水平,pLGFD9-12嵌合克隆衍生病毒在DL细胞上复制水平略高于FDD细胞.此结果为进一步深入研究LTR对马传染性贫血病毒复制水平和毒力的影响奠定了基础.     

GSK-3在阿尔茨海默病样细胞骨架蛋白过度磷酸化中的作用(英)

神经原纤维缠结是阿尔茨海默病（Alzheimer disease, AD）的特征性病理改变.蛋白激酶和蛋白磷酸酯酶失衡可导致骨架蛋白的异常过度磷酸化,而异常过度磷酸化的tau 和神经丝 (neurofilament, NF) 是神经原纤维缠结的组成部分.在众多激酶中,糖原合酶激酶-3（glycogen synthase kinase-3,GSK-3）可能是AD神经退行性变起重要作用.为深入探讨GSK-3在AD样神经退行性变中的作用,以磷酯酰肌醇三磷酸激酶（phosphatidylinositol 3-kinase,PI3K）的特异性抑制剂渥曼青霉素（wortmannin,WT）处理野生型鼠成神经瘤细胞株（wild type mouse neuroblastoma cell lines, N2a wt）,系统观察WT处理N2a wt不同时间点（1 h、3 h、6 h）细胞代谢率、细胞形态、细胞骨架蛋白tau和NF的磷酸化状态改变以及细胞的命运,并分析了GSK-3活性与上述参数改变之间的相关性.结果发现：1 μmol/L WT处理细胞1 h,GSK-3活性与未经WT处理的对照组相比明显增高,并伴有Ser9磷酸化的GSK-3水平的降低; NF磷酸化程度增强,tau在Ser198/Ser199/Ser202位点的磷酸化增强. 1 μmol/L WT处理细胞3 h,GSK-3活性与对照组和处理1 h 组相比明显下降,NF磷酸化程度较1 h降低,但仍高于正常水平.1 μmol/L WT处理细胞6 h,细胞形态、GSK-3活性、Ser9磷酸化形式的GSK-3β的表达、NF磷酸化程度与对照组相比均无明显改变.WT呈剂量依赖性降低细胞代谢率.1 μmol/L WT处理细胞1 h和3 h导致细胞变圆,突起变短甚至消失.1 μmol/L WT处理细胞1 h,用TUNEL法和电子显微镜技术未观察到细胞凋亡.研究结果提示：在N2a细胞中过度激活GSK-3可导致神经细丝和tau蛋白的AD样过度磷酸化,从而引起神经细胞的AD样退行性变.     

Inhibition of mitochondrial complex I improves glucose metabolism independently of AMPK activation

Accumulating evidences showed metformin and berberine, well‐known glucose‐lowering agents, were able to inhibit mitochondrial electron transport chain at complex I. In this study, we aimed to explore the antihyperglycaemic effect of complex I inhibition. Rotenone, amobarbital and gene silence of NDUFA13 were used to inhibit complex I. Intraperitoneal glucose tolerance test and insulin tolerance test were performed in db/db mice. Lactate release and glucose consumption were measured to investigate glucose metabolism in HepG2 hepatocytes and C2C12 myotubes. Glucose output was measured in primary hepatocytes. Compound C and adenoviruses expressing dominant negative AMP‐activated protein kinase (AMPK) α1/2 were exploited to inactivate AMPK pathway. Cellular NAD⁺/NADH ratio was assayed to evaluate energy transforming and redox state. Rotenone ameliorated hyperglycaemia and insulin resistance in db/db mice. It induced glucose consumption and glycolysis and reduced hepatic glucose output. Rotenone also activated AMPK. Furthermore, it remained effective with AMPK inactivation. The enhanced glycolysis and repressed gluconeogenesis correlated with a reduction in cellular NAD⁺/NADH ratio, which resulted from complex I suppression. Amobarbital, another representative complex I inhibitor, stimulated glucose consumption and decreased hepatic glucose output in vitro, too. Similar changes were observed while expression of NDUFA13, a subunit of complex I, was knocked down with gene silencing. These findings reveal mitochondrial complex I emerges as a key drug target for diabetes treatment. Inhibition of complex I improves glucose homoeostasis via non‐AMPK pathway, which may relate to the suppression of the cellular NAD⁺/NADH ratio.     

禽病原性大肠杆菌1型菌毛的分离与鉴定

以旋涡混合法使禽病原性大肠杆菌分离株566、1794和TK3菌毛脱落,经硫酸铵沉淀、透析后进行蔗糖密度梯度离心,收集密度为110至115g/cm3的蛋白带,经SDSPAGE测定,3株菌菌毛蛋白的分子量分别在175、170和170kD;提纯菌毛保留了甘露糖敏感性凝集豚鼠红细胞的能力,证明它们为1型菌毛;从1794株提取的1型菌毛免疫BALB/C小鼠产生的高免血清在Western blot中与3个菌株的相应菌毛蛋白均呈阳性反应。上述结果表明,受检的3株禽病原性大肠杆菌均表达了1型菌毛,其分子量在175～170kD之间,3个菌株的1型菌毛间具有较强的抗原相关性。     

利用cDNA-RDA技术研究BXSB狼疮小鼠差异表达的基因

利用近年来发展起来的代表性差异分析cDNA-RDA(cDNA-representational difference analysis)技术筛选BXSB红斑狼疮小鼠发病相关基因.发现了3个新的表达序列标签(EST)片段,在GenBank中的登录号分别为AF060113, AF060111, AF060110,同时发现了一些已知与自身免疫病相关的基因如逆转录病毒衣壳蛋白、Line-1逆转录酶等.通过RDA技术可能发现系统性红斑狼疮发病相关新基因,为自身免疫病的理论研究提供新的思路和方法.     

Targeting cleavage and polyadenylation specific factor 1 via shRNA inhibits cell proliferation in human ovarian cancer

Cleavage and polyadenylation specificity factor 1 (CPSF1), a member of CPSF complex, has been reported to play a key role in pre-mRNA 3′-end formation, but its possible role in ovarian cancer remains unclear. In the present study, we found the mRNA level of CPSF1 was overexpressed in ovarian cancer tissues using Oncomine Cancer Microarray database. Then the loss-of-function assays, including CCK-8, colony formation and flow cytometry assays, were performed to determine the effects of CPSF1 on cell viability, proliferation, cell cycle and apoptosis of human ovarian cancer cell lines (SKOV-3 and OVCAR-3). The results indicated that depletion of CPSF1 suppressed cell viability, impaired colony formation ability, induced cell cycle arrest at G0/G1 phase and promoted cell apoptosis in ovarian cancer cells. Furthermore, knockdown of CPSF1 upregulated the expression of cleaved caspase-3 and PARP and downregulated CDK4/cyclin D1 expression. These data suggested that CPSF1 could promote ovarian cancer cell growth and proliferation in vitro and its depletion might serve as a potential therapeutic target for human ovarian cancer.     

57份苎麻种质资源主要农艺性状及纤维品质鉴定评价

对57份苎麻种质的主要农艺性状和纤维品质性状进行了鉴定评价。结果筛选出高产苎麻种质10份(原麻产量≥2000 kg/hm2),纤维品质优良的种质7份(纤维细度≥2000 m/g),鲜皮出麻率≥12.5%的苎麻种质20份,炼折率超过65%的苎麻种质有10份,按照苎麻优异种质评价规范,满足高产和出麻率高2个优良指标的种质7份,满足优质和鲜皮出麻率高2个优良指标的种质1份。     

黑果枸杞的组织培养

1植物名称黑果枸杞(Lycium ruthenicum Murr.). 2材料类别带腋芽的嫩茎段. 3培养条件诱导分化培养基:(1)MS 6-BA 1.0mg·L-1(单位下同) NAA 0.2,(2)MS 6-BA 0.5 NAA0.2;芽增殖培养基:(3)MS 6-BA 0.5 NAA 0.1(4)MS 6-BA 0.5 NA 0.2;生根培养基为:(5)1/2MS NAA 0.1.以上诱导分化及增殖培养基均加2.5%蔗糖和0.8%琼脂,生根培养基的蔗糖和琼脂量减半,pH 5.8～6.0.培养温度为23～25℃,光照时间10～12 h·d-1,光照度为2000 1x.