夏枯草PvGGPPS基因的克隆和诱导表达分析

为探究夏枯草中GGPPS基因的生物学特性及功能,该文在夏枯草转录组测序的基础上设计特异性引物,采用逆转录PCR技术获得夏枯草中GGPPS基因的全长核苷酸序列,并进行生物信息学分析;采用qPCR法分析PvGGPPS基因在不同外源性物质诱导下在夏枯草果穗中的表达量以及该基因在夏枯草不同组织中的表达量。结果表明:PvGGPPS基因开放阅读框1 092 bp,编码363个氨基酸,理论分子量为38 815.68 D,等电点为5.69。PvGGPPS蛋白具有异戊烯基焦磷酸合酶家族的特征结构域。系统进化树表明PvGGPPS蛋白与丹参、毛喉鞘蕊花GGPPS蛋白具有较高的亲缘关系。qPCR分析表明,PvGGPPS基因在叶中表达量高于果穗及茎。对果穗施加7种外源性物质处理24 h后,GA3处理组该基因表达量升高。PvGGPPS基因在夏枯草不同组织中表达量差异较大,且受外源物质诱导表达。该研究结果为进一步研究PvGGPPS基因对夏枯草萜类成分合成途径中的功能及表达调控奠定基础。     

夏枯草多糖的分离纯化与抗氧化活性研究

对夏枯草多糖(PP2)的分离纯化方法作了探讨,通过正交试验,筛选出多糖提取条件的最佳组合。采用纸层析、琼脂糖凝胶电泳和紫外分光光度法对多糖纯度进行鉴定。通过气相色谱仪对多糖的单糖组成进行分析。通过体外化学模拟,研究夏枯草多糖的抗氧化性。实验表明,夏枯草多糖对O2-.、.OH二种自由基及亚硝酸根离子具有一定的清除能力,对R.自由基的清除较弱,具有防止膜脂质过氧化,减少红细胞溶血和降低脂质过氧化产物丙二醛的生成量的作用。     

Effects of the winter beechmast crop on bird occurrence in British gardens

Capsule Woodland birds were significantly less likely to occur in gardens in years of high beechmast crop.

Aim To test the hypothesis that woodland species that feed on beechmast will have significantly lower occurrence rates at garden feeders in mast years.

Methods Weekly winter occurrence rates at garden feeders between 1970/71 and 1999/2000 for 40 species were analysed in relation to annual beechmast abundance, classified into low, medium and high years. A repeated-measures logistic regression model was used to assess whether beechmast abundance explained further significant variation additional to underlying seasonal and annual trends.

Results Seven species that commonly feed on beechmast showed significantly lower occurrence in gardens in years of highest beechmast abundance: Great Spotted Woodpecker Dendrocopos major, Woodpigeon Columba palumbus, Great Tit Parus major, Coal Tit Periparus ater, Nuthatch Sitta europaea, Jay Garrulus glandarius and Chaffinch Fringilla coelebs. Blackbird Turdus merula and Siskin Carduelis spinus, which showed similar significant patterns, are likely to take beechmast as elements of their diet. Pied Wagtail Motacilla alba was the only insectivorous species to show significant effects, but occurrence was lowest in years of intermediate beechmast abundance. For the latter species, this may have been due to confounding effects of temperature, but there were no such confounding effects of either temperature, or the number of bird feeders provided in gardens, for the other nine species.

Conclusion Use of artificial food sources by birds in gardens is influenced by resources in the surrounding countryside, suggesting that food provided in gardens may play a significant part in the population dynamics of these species, that population monitoring without consideration of the garden habitat may be deficient, and that volunteer-based garden bird recording may provide data that can be used as an indicator of changes in the wider countryside.     

Inhibition of estrogen signaling through depletion of estrogen receptor alpha by ursolic acid and betulinic acid from Prunella vulgaris var. lilacina

Extracts of Prunella vulgaris have been shown to exert antiestrogenic effects. To identify the compounds responsible for these actions, we isolated the constituents of P. vulgaris and tested their individual antiestrogenic effects. Rosmarinic acid, caffeic acid, ursolic acid (UA), oleanolic acid, hyperoside, rutin and betulinic acid (BA) were isolated from the flower stalks of P. vulgaris var. lilacina Nakai (Labiatae). Among these constituents, UA and BA showed significant antiestrogenic effects, measured as a decrease in the mRNA level of GREB1, an estrogen-responsive protein; the effects of BA were stronger than those of UA. UA and BA were capable of suppressing estrogen response element (ERE)-dependent luciferase activity and expression of estrogen-responsive genes in response to exposure to estradiol, further supporting the suppressive role of these compounds in estrogen-induced signaling. However, neither UA nor BA was capable of suppressing estrogen signaling in cells ectopically overexpressing estrogen receptor α (ERα). Furthermore, both mRNA and protein levels of ERα were reduced by treatment with UA or BA, suggesting that UA and BA inhibit estrogen signaling by suppressing the expression of ERα. Interestingly, both compounds enhanced prostate-specific antigen promoter activity. Collectively, these findings demonstrate that UA and BA are responsible for the antiestrogenic effects of P. vulgaris and suggest their potential use as therapeutic agents against estrogen-dependent tumors.     

夏枯草药材和种植土壤中农药及重金属残留分析

采用气相色谱及ICP-AES法测定了安徽庐江和江苏洪泽2个种植基地的土壤和夏枯草(Prunella vulgaris L.)果穗及全草中有机氯农药及重金属含量,并根据污染指数和相关标准对土壤及药材的安全性进行了评价.测定结果表明:来源于2个基地的土壤及药材中有机氯农药及重金属含量有明显差异.庐江产果穗和全草中Pb、Cd、Cu、Cr、As及BHC含量分别为3.361和3.953、0.172和0.190、8.258和7.722、3.423和2.658、0.284和0.355、0.003和0.004 mg·kg-1,Hg和DDT未检出;洪泽产果穗和全草中Pb、Cd、Cu、Cr、Hg及BHC含量分别为2.399和1.558、0.155和0.111、7.682和6.756、4.259和3.801、0.077和0.102、0.003和0.006 mg·kg-1,As未检出,果穗中也未检出DDT.庐江基地土壤中Cd、Cu、Cr、As、Hg、BHC和DDT含量分别为0.001、12.943、47.417、1.008、0.003、0.003和0.002 mg·kg-1,Pb未检出;洪泽基地土壤中Pb、Cd、Cu、Cr、As、Hg和BHC含量分别为3.443、0.002、18.655、63.385、3.701、0.141和0.004 mg·kg-1,DDT未检出.比较结果表明:夏枯草果穗中重金属残留量均高于全草,但均低于国家限量标准;土壤中有机氯农药及重金属单项污染指数均小于1,且庐江和洪泽基地土壤的综合污染指数分别为0.286和0.399,因此,土壤污染等级属安全级且污染水平为清洁级.     

The multicomponent phytopharmaceutical SKI306X inhibits in vitro cartilage degradation and the production of inflammatory mediators

Clinical studies have demonstrated that SKI306X, a purified preparation of three medicinal plants, relieves joint pain and improves functionality in osteoarthritis patients. To study the biological action of SKI306X, bovine cartilage explants and human peripheral blood mononuclear cells (PBMC) were stimulated with IL-1β and lipopolysaccharide (LPS) respectively, in the presence or absence of SKI306X and its individual composites. All tested compounds inhibited dose-dependently IL-1β-induced proteoglycan release and nitric oxide production by cartilage, indicating cartilage protective activity. SKI306X and two of its compounds inhibited PGE₂, TNF- and IL-1β production by LPS-stimulated PBMC, indicating anti-inflammatory activity. These results demonstrate that the biological effect of SKI306X is at least bipartite: (1) cartilage protective and (2) anti-inflammatory. The observed anti-inflammatory effects may provide an explanation for the outcome of the clinical studies. Long-term clinical trails are necessary to elucidate whether the in vitro cartilage protective activity results in disease-modifying effects.     

夏枯草诱导人甲状腺乳头状癌细胞K1增殖和凋亡的影响及其作用机制

目的:探索夏枯草对人甲状腺乳头状癌细胞(K1)增殖和诱导K1细胞凋亡的影响及其可能的作用机制。方法:采用MTT比色法测定2~200 mg/mL夏枯草作用24 h对K1细胞增殖的影响;采用Hoechst染色法和流式细胞术(Annexin V-FITC/PI联合标记)观察0.3、0.6、1.2、2.4、4.8 mg/mL夏枯草作用24 h K1细胞凋亡的影响;采用蛋白质免疫印迹法(Western blotting)测定1.2、2.4、4.8 mg/mL夏枯草作用24h对K1细胞凋亡相关蛋白表达的影响。结果:在2~200 mg/mL的浓度范围内夏枯草作用24 h对K1细胞增殖有明显抑制作用(P0.05),IC50值为2.427 mg/mL。流式细胞术检测结果显示夏枯草可诱导K1细胞凋亡,2.4、4.8 mg/mL组K1细胞总凋亡率分别为(11.35±0.92)%、(44.57±3.07)%,与对照组相比明显升高(P0.05);与对照组相比,1.2、2.4、4.8 mg/mL夏枯草作用24 h胞浆内凋亡蛋白Bax、细胞色素C(Cyto C)、Caspase-9、活化的Caspase-3(Cleaved Caspase-3)表达增多。结论:夏枯草能抑制人甲状腺癌K1细胞增殖并诱导其凋亡,其机制可能与活化线粒体凋亡通路有关。     

秦岭发现黑喉岩鹨

2001年3月20日在秦岭主脊采集到黑喉岩鹨(Prunella atrogularis)1只,为秦岭鸟类新纪录.本文对其形态特征及采集地和生境进行了描述,对其居留型进行了分析.     

棕眉山岩鹨的代谢产热特征的初步研究

实验采用封闭式流体压力呼吸计和数字式温度计 ,分别测定了环境温度 (Ta)在 5～ 35℃范围内棕眉山岩鹨 (Prunellamontanella)的耗氧量和体温 (Tb) ,由此计算出代谢率 (MR)、热传导率 (C)和代谢预期比与热传导预期比 (F)等值 ,以探讨该种的代谢产热特征。结果表明 :棕眉山岩鹨在Ta 为 5～ 35℃时 ,Tb 基本维持恒定 (为 4 1 2 5± 0 12℃ ) ,MR的最低值为 4 2 7± 0 0 7mLO2 /(g·h) ;Ta 在 5～ 2 4℃时 ,MR与之负相关 :MR [mLO2 /(g·h) ]=10 39- 0 2 4Ta℃ ;Ta 在 5～ 2 6 5℃时 ,C最低且基本保持恒定 ,平均为 0 2 6±0 0 0mLO2 /(g·h·℃ ) ;Ta 在 2 4～ 30℃时 ,F值为 1 0 5 ,表明该种具有较好的体温调节能力。与热带地区鸟类比较 ,棕眉山岩鹨具较高的MR和Tb,稍高的C ,符合北方小型鸟类的代谢特点 ,能较好的适应冬寒冷、夏炎热的环境。     

甘肃莲花山自然保护区栗背岩鹨的繁殖生态初步观察

2001年7月在甘肃省莲花山自然保护区,对栗背岩鹨的繁殖习性作了考察,详细记述了这种鸟的4个巢,并报道了该鸟的声谱及其幼鸟的形态。