雪糕中16种反式脂肪酸的测定

建立气相色谱测定雪糕中反式脂肪酸含量的方法.2.0 g雪糕样品中加入8 mL浓盐酸和10 mL无水乙醇进行前处理,经SP-2560毛细管色谱柱分离,气相色谱仪-氢火焰离子化检测器检测,面积归一化法计算组分含量.结果表明:在优化的色谱条件下,方法的检出限和定量限分别为0.012％和0.024％;实现了反式脂肪酸甲酯与顺式...     

High‐level exogenous trans10, cis12 conjugated linoleic acid plays an anti‐lipogenesis role in bovine mammary epithelial cells

Primary bovine mammary epithelial cells (BMECs) were treated by 0, 37.5, 75, 112.5, 150 μmol/L trans10, cis12 conjugated linoleic acid (CLA) to evaluate the effects of different level trans10, cis12 CLA on lipogenesis in BMEC. Addition of 75–150 μmol/L trans10, cis12 CLA reduced significantly the triacylglycerol (TAG) content (P < 0.05), but did not have inhibiting action on cell proliferation (P > 0.05). Treatment with 150 μmol/L trans10, cis12 CLA for 48 h resulted in a 17.1% reduction (P < 0.0001) of medium chain fatty acids (MCFA, C14 < C < C16), a 26.5% reduction (P < 0.0001) of unsaturated fatty acids (UFA) and a corresponding reduction of the mRNA abundance of acetyl coenzyme A (acetylCoA) carboxylase (ACC) (P = 0.046), fatty acid synthase (FAS) (P = 0.017) and stearoylCoA desaturase1 (SCD1) (P = 0.002). Another finding was that trans10, cis12 CLA elevated expression of diacylglycerol acyltransferase2 (DGAT2) (P = 0.020) and long chain acylCoA synthetases (ACSL) (P = 0.032). In conclusion, higher trans10, cis12 CLA, not low trans10, cis12 CLA, inhibited milk fat synthesis and changed fatty acid composition by regulating the expression of FAS, ACC, SCD1, DGAT2 and ACSL.     

Carotenoid composition of a New Zealand (Evechinus chloroticus) and an Australian (Heliocidaris erythrogramma) sea urchin in relation to gonad colour

Sea urchins are commercially harvested for their edible gonads, also known as roe or uni. Most species, particularly those from the Northern hemisphere are reported to exhibit limited variation in gonad colour, which is proposed to be due to carotenoid pigments. The commercially harvested species from New Zealand, Evechinus chloroticus, and from Australia, Heliocidaris erythrogramma, however, exhibit considerable gonad colour variation, ranging from a culinary desirable light yellow/orange to an undesirable brown. Gonads from E. chloroticus and H. erythrogramma were characterized spectrophotometrically and the carotenoid content extracted and analysed by reversed‐phase high‐performance liquid chromatography. The main carotenoid found in the gonads of both species was 9′‐cis‐echinenone, with lower levels of all‐trans‐echinenone, lutein and isozeaxanthin. Carotenoid composition contributed to the yellow/orange colour components of the gonads, but was not correlated with the darker roe coloration observed in some E. chloroticus individuals. Our results highlight important considerations relating sea urchin gonad coloration to carotenoid content and as such will provide useful information for the future aquaculture and processing of these two species.     

橡胶树延伸因子cDNA及其5′端启动子区域序列的分离与分析

橡胶延伸因子REF(Rubber Elongation Factor)是橡胶生物合成中最重要的酶之一, 是胶乳中主要的蛋白质. 作者利用已发表的REF cDNA序列设计并合成引物. 通过提取胶乳总RNA用RT法合成cDNA后, 通过PCR技术扩增并克隆了REF cDNA序列. 序列测定结果表明, 所克隆的REF cDNA编码区序列全长414个碱基, 编码138个氨基酸, 3′端非编码区     

Thermal decomposition and isomerization of cis-permethrin and beta-cypermethrin in the solid phase

The stability to heart of cis-permethrin and beta-cypermethrin in the solid phase was studied and the decomposition products identified. Samples heated at 210 degrees C in an oven in the dark showed that, in the absence of potassium chlorate (the salt present in smoke-generating formulations of these pyrethroids), cis-permethrin was not isomerized, although in the presence of that salt, decomposition was greater and thermal isomerization occurred. Other salts of the type KXO3 or NaXO3, with X being halogen or nitrogen, also led to a considerable thermal isomerization. Heating the insecticides in solution in the presence of potassium chlorate did not produce isomerization in any of the solvents assayed. Salt-catalysed thermal cis-trans isomerization was also found for other pyrethroids derived from permethrinic or deltamethrinic acid but not for those derived from chrysanthemic acid. The main thermal degradation processes of cis-permethrin and beta-cypermethrin decomposition when potassium chlorate was present were cyclopropane isomerization, ester cleavage and subsequent oxidation of the resulting products. Permethrinic acid, 3-phenoxybenzyle chloride, alcohol, aldehyde and acid were identified in both cases, as well as 3-phenoxybenzyl cyanide from beta-cypermethrin. A similar decomposition pattern occurred after combustion of pyrethroid fumigant formulations.     

蜜蜂球囊菌中长链非编码RNA的调控作用

【目的】长链非编码RNA（long non-coding RNA,lncRNA）是一类二、三级结构高度保守,长度>200 nt且不具蛋白编码能力的RNA,在转录和转录后水平广泛参与调控剂量补偿、细胞分化和生长发育等生命活动。本研究基于前期获得的蜜蜂球囊菌（Ascosphaera apis,简称球囊菌）菌丝和孢子混合样品的高质量lncRNA组学数据进行球囊菌lncRNA的顺式（cis）作用、反义lncRNA（antisense lncRNA）作用和竞争性内源RNA（competing endogenous RNA,ceRNA）作用的分析和探讨,以期揭示lncRNA在球囊菌中的潜在功能。【方法】基于lncRNA基因在染色体上的位置,预测lncRNA上下游100 kb以内的蛋白编码基因;使用Blast软件将上下游基因比对到GO和KEGG数据库,以获得功能和通路注释。利用LncTar软件对反义lncRNA的靶mRNA进行预测,并使用Blast软件将上述靶mRNA比对到KEGG和eggNOG数据库。利用TargetFinder软件预测lncRNA靶向结合的miRNA及miRNA靶向结合的mRNA,根据靶向结合关系建立lncRNA-miRNA和lncRNA-miRNA-mRNA调控网络,进而通过Cytoscape v3.7.1软件进行调控网络的可视化。利用RT-PCR对调控网络中的lncRNA、靶miRNA和靶mRNA进行表达验证。【结果】共预测出371个lncRNA的5 852个上下游基因,可注释到细胞进程、代谢进程和应激反应等48个功能条目,以及新陈代谢途径、次生代谢产物的生物合成和抗生素的生物合成等121条通路,表明部分lncRNA可通过顺式作用调节上下游基因的表达,从而参与调控球囊菌的生长发育及物质能量代谢等基础生命活动。球囊菌的7个lncRNA与7个靶mRNA存在序列互补关系,其中5个mRNA在eggNOG数据库中仅注释为假定蛋白,仅gene3444在KEGG数据库注释为核孔复合体蛋白An-Nup120和假定蛋白,表明上述1个反义lncRNA可能参与调控核孔复合体蛋白的生物合成等生物学过程。此外,共预测出227个lncRNA与73个miRNA之间存在靶向结合关系,其中多数lncRNA(79.02%)仅能结合1—2个miRNA,部分miRNA可被多个lncRNA靶向结合;进一步构建氧化磷酸化通路和MAPK信号通路相关的lncRNA-miRNA-mRNA调控网络,分析结果显示氧化磷酸化通路相关的222个lncRNA靶向78个miRNA及50个mRNA,MAPK信号通路相关的222个lncRNA靶向76个miRNA及46个mRNA,表明部分lncRNA通过ceRNA作用调控此两条通路,从而影响球囊菌的能量合成、环境适应以及生长发育等过程。【结论】部分lncRNA可能通过顺式作用和ceRNA作用调节球囊菌的生长发育、物质能量代谢以及环境适应等生物学过程;MSTRG.5393.1可能作为反义lncRNA调控球囊菌的核孔复合体的蛋白合成。     

番茄LeNCED1基因5′-侧翼序列克隆及LeNCED1p:GUS融合基因构建

从番茄基因组中克隆LeNCED1基因5′-侧翼1456 bp调控序列,经生物信息学分析表明,LeNCED1基因上游调控序列中存在响应昼夜节律的顺式作用元件Circadian、光响应元件Box I、逆境胁迫响应元件(TC-rich repeats)、ABA响应元件ABRE等,构建了LeNCED1基因上游调控序列与报告基因GUS融合的植物双元表达载体pLeNCED1p:GUS。     

Pin1 inhibitor juglone induces apoptosis in human cervical cancer SiHa cells

AIM: To explore the effect of peptidyl-prolyl cis/trans isomerase (Pin1) inhibitor juglone on apoptosis of human cervical cancer SiHa cells. METHODS: Cultured SiHa cells were incubated with juglone at concentrations of 10, 20, 50, 80 and 100 μmol/L for 24 h. The SiHa cell activity was detected by methyl thiazolyl tetrazolium (MTT) assay. The cell apoptosis was analyzed by flow cytometry with Hoechst 33258 staining. The protein levels of cleaved caspase-3,8,9 and PTEN was determined by Western blotting. RESULTS: In different doses of juglone groups, the SiHa cell growth was greatly inhibited (P<0.05) in a dose-dependent manner as compared with control group. The IC₅₀ of juglone was 20.4 μmol/L. After treatment with juglone at concentration of 20 μmol/L for 12 h, the apoptosis of SiHa cells was induced, and the typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus, nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining. The early apoptotic rate was increased significantly as compared with the control. The protein levels of cleaved caspase-3, 8, 9 and PTEN were also increased significantly as compared with control group. CONCLUSION: Juglone significantly inhibits the cell activity and induces the apoptosis of SiHa cells in vitro by inhibiting the caspase pathway and increasing the expression of anti-oncogene.     

Stilbene Levels in the Tissue and Juice of Muscadine Grapes (Vitis rotundifolia Michx.)

Abstract

Stilbenes are secondary metabolites of a class of non-flavanoid phenolics found in certain species of Vitis. This class of compounds includes resveratrol and a resveratrol glucoside called piceid that exists in trans and cis forms. This study was undertaken to determine the cis and trans forms of both resveratrol and piceid in the skin, pulp, seeds, and juice of nine cultivars of muscadine grapes and three cultivars of Vitis labruscana. Juice samples of the muscadine cultivars ‘Fry’, ‘Hunt’, ‘Magnolia’, ‘Watergate’, ‘Carlos’, ‘Noble’, and ‘Sweet Jenny’, and the V. labruscana cultivars ‘Albermarle’, ‘Miss Blanc’, ‘MidSouth’ and ‘Miss Blue’ were harvested at the full ripe stage and divided into two sub-samples. One subsample was used to extract juice and the other was divided into skins, pulp, and seeds. Sample analysis was performed using HPLC with a UV detector. Sample chromatagrams were compared with those of known standards for quantification. ‘Carlos’ and ‘Magnolia’ had the greatest total stilbene concentration in skin tissue and had bronze skin containing higher concentrations of total stilbenes than any of the black-skinned cultivars tested. Only one bronze skin cultivar, ‘Sweet Jenny’, had lower stilbenes than the dark skin cultivars. Piceid, cis piceid and resveratrol were found in the skins of all cultivars. ‘Carlos’ ‘Magnolia’, ‘Fry’ and ‘Albermarle’ muscadines contained cis resveratrol in the skins of the fruit. ‘Watergate’ was the only cultivar in which resveratrol was detected in seed tissue. ‘Sweet Jenny’ had the highest levels of cis piceid in seed tissue. ‘Albermarle’, ‘Carlos’ and ‘Sweet Jenny’ had significantly greater levels of cis picied in pulp tissue than all other cultivars.