栗柄金粉蕨的黄酮类成分

从栗柄金粉蕨（ Ｏｎｙｃｈｉｕｍ ｌｕｃｉｄｕｍ ） 地上部分的甲醇抽提物中分到１０ 个成分, 经详细的一维、二维核磁数据分析, 它们被鉴定为： 木犀草甙（１） , ３ , ７ － 二甲基槲皮素（２） , 高山甙Ｂ （３） , 金粉蕨素（４） , 栗柄醇（５） , 金粉蕨醇Ｂ（６） , β－ 谷甾醇（７） , 胡萝卜甙（８） ,齐墩果酸（９） 和蔗糖（１０） 。栗柄醇系新成分。１ , ２, ９ 系首次由金粉蕨属分到。     

新疆圆柏化学成分研究

从新疆圆柏枝叶中分离得到9个化合物,经理化性质和光谱解析分别为扁柏双黄酮（hinokiflavone,1）、芹菜甙元（apigenin,2）、鬼臼毒素（podophyllotoxin,3）、苦松甙（coniferin,4）、肉豆蔻酸丙酯（propionic acetate myristic acid,5）、β-谷甾醇（β-sitosterol,6）、圆柏醇（sabinal,7）、反式璎珞酸（trans-communic acid,8）、蔗糖（sucrose,9）,其中化合物1～4为首次从该植物中提取分离得到.     

高山金粉蕨的黄酮类成分

从高山金粉蕨(Onychiumcontigqum)地上部分的甲醇抽提物中分到8个成分:高山金粉蕨甲甙(1),高山金粉蕨乙甙(2),金粉蕨素(3),反式桂皮酸(4),瓦利甙(5),β-谷甾醇(6),胡萝卜甙(7)和蔗糖(8)。高山金粉蕨甲甙和乙甙是新成分,反式桂皮酸系首次从金粉蕨属中分到。化学结构用一维和二维核磁共振技术确定。     

迷迭香水溶性物质及抗菌活性研究

采用硅胶柱色谱等方法对迷迭香水溶性提取物进行分离纯化,用^13C NMR、^1H NMR、Ms等方法鉴定出3个化合物：迷迭香酸（1）、香叶木素-7-O-β-D-葡萄糖醛酸甙（2）、木犀草素（3）,其中香叶木素-7-O-β-D-葡萄糖醛酸甙为首次从该植物中发现。对化合物2进行体外抑菌实验,结果为：对金色葡萄球菌和枯草芽孢杆菌均有一定的抑菌作用。     

坚龙胆中的一个新裂环烯醚萜甙

从坚龙胆（Gentiana rigescens）的根中分离得到1个新的裂环烯醚萜甙,命名为坚龙胆甙A（1）,以及8个已知化合物：龙胆苦甙（2）,6＇-O-β-D-吡喃葡萄糖基-龙胆苦甙（3）,马钱子酸（4）,6＇-O-β-D-吡喃葡萄糖基．马钱子酸（5）,獐牙菜甙（6）,2＇-（邻,间-二羟基苯甲酰基）．獐牙菜甙（7）,獐牙菜苦甙（8）,四乙酰开联番木鳖甙（9）。它们的化学结构通过现代波谱解析得以鉴定。化合物3,5和9为首次从坚龙胆中分离得到。     

栗柄金粉蕨的黄酮成分

从湖南省江华瑶族自治县产的栗柄金粉蕨（Ｏｎｙｃｈｉｕｍ ｌｕｃｉｄｕｍ）的地上部分分离得到４个黄酮化合物,经光谱鉴定为：木犀草素－７－葡粮甙（１）,３,７－二甲氧基槲皮黄素（２）,槲皮黄素－３－葡糖甙（３）和金粉蕨素（４）,均为首次从该植物中获得。     

Flavonoids of Onychium lucidum

从栗柄金粉蕨(Onychium lucidum)地上部分的甲醇抽提物中分到10个成分,经详细的一维、二维核磁数据分析,它们被鉴定为木犀草甙(1),3,7-二甲基槲皮素(2),高山甙B(3),金粉蕨素(4),栗柄醇(5),金粉蕨醇B(6),β-谷甾醇(7),胡萝卜甙(8),齐墩果酸(9)和蔗糖(10)。栗柄醇系新成分。1,2,9系首次由金粉蕨属分到。     

藏药黑边假龙胆的化学研究

龙胆科植物黑边假龙胆（Ｇｅｎｔｉａｎｅｌｌａａｚｕｒｅａ（Ｂｕｎｇｅ）Ｈｏｌｕｂ）是治疗肝胆湿热的１种藏药,从西藏日喀则产的全草中分离得到９个化合物,其中１个Ｃ一甙黄酮,异荭草素（ｉｓｏｏｒｉｅｎｔｉｎ）;１个山酮甙,獐牙菜酚甙（ｓｗｅｒｔｉａｎｏｌｉｎ）;１个木质体甙,橄榄脂素４一Ｏ一β－Ｄ－葡萄糖甙（ｏｌｉｖｉｌ４一Ｏ一β一ｇｌｕｃｏｓｉｄｅ）;３个环烯醚萜甙,龙胆苦甙（ｇｅｎｔｉｏｐｉｃｒｏｓｉｄｅ）,獐牙菜甙（ｓｗｅｒｔｉａｍａｒｉｎ）和马钱子酸（ｌｏｇａｎｉｃａｃｉｄ）;１个三萜,齐敦果酸（ｌｏｅａｎｏｌｉｃａｃｉｄ）１个甾醇,β谷甾醇葡萄糖甙（ｄａｕｃｏｓｔｅｒｏｌ）以及１个长链脂肪醇,三十烷醇（ｎ一１一ｔｒｉａｃｏｎｔａｎｏｌ）。它们的结构通过光谱和化学的方法得到了鉴定。上述结果,从次生代谢产物的角度为假龙胆属在系统分类上与龙胆属和獐牙菜属有着密切的联系提供了旁证。     

抱茎獐牙菜的San酮甙和黄酮甙

从龙胆科植物抱茎獐牙菜（Swertia franchtiana H.Smith)中分得3种San酮甙,1种黄酮甙和1种三萜酸,通过光谱和化学等方法,分别鉴定为当药醇甙,当药黄素,芒果甙,1－O－[β－D－吡喃木糖－（1－6）－β－D－吡喃葡萄糖],－3,5－二甲氧基San酮（XI）和齐墩果酸,其中晶XI为一新化合物。     

茉莉花的化学成分

从药用植物茉莉花(Jasminum sambac(L．)Ait．)花蕾中分离到9个化合物,通过波谱分析并与已知化合物数据对照,分别鉴定为：苄基-O-β-D-葡萄吡喃糖甙(1),苄基-O-β-D-木吡喃糖基(1→6)-β-D-葡萄吡喃糖甙(2),tetraol(3),molihuaoside D(4),sarnhcoside A(5),sambacoside E(6),芦丁(rutin)(7),山奈酚-3-O-α-L鼠李吡喃糖基(1-2)[α-L鼠李吡喃糖基(1-6)]-β-D半乳吡喃糖甙(8),斛皮素-3-O-α-L鼠李吡喃糖基(1→2)[α-L鼠李吡喃糖基(1→6)]-β-D-半乳吡喃糖甙(9)。     

Comparison of three fluorescent CYP3A substrates in two vertebrate models: pig and Atlantic salmon

We investigated in vitro inhibitory effects of ketoconazole (KTZ) on cytochrome P450 activity in microsomes from pigs and Atlantic salmon. The following enzymatic reactions were studied: 7-benzyloxyresorufin and 7-ethoxyresorufin O-dealkylation (BROD and EROD, respectively), 7-benzyloxy-4-trifluoromethylcoumarin O-debenzylation (BFCOD) and 7-benzyloxyquinoline O-debenzylation (BQOD). KTZ was a potent non-competitive inhibitor of BROD and BQOD in the microsomes from pigs, whereas in the microsomes from Atlantic salmon, these reactions were competitively inhibited by KTZ. BFCOD activity was inhibited by KTZ in a non-competitive manner in both species. KTZ non-competitively inhibited EROD in Atlantic salmon, but not in porcine microsomes. The activity of BROD and BQOD was higher in male than that in female pigs, but the activity of BFCOD showed no sex-related differences.     

人蛋白激酶CK2α′亚基在大肠杆菌中的克隆、表达及其重组蛋白的纯化与性质鉴定

通过RT PCR从HL 6 0细胞获得人蛋白激酶CK2α′亚基编码区cDNA ,将NdeⅠ HindⅢ双酶切的PCR产物和pT7 7表达载体进行定向克隆、细菌转化、电泳初筛和限制性酶切分析鉴定 .随机挑选阳性克隆进行DNA测序确证 ,筛选含与已知序列完全相符的重组质粒 (命名为pTCKA′) .将其转化BL2 1(DE3)菌 ,IPTG诱导后未见高效特异表达 .然后将人CK2α′cDNA亚克隆至GST融合蛋白表达载体 ,经同样转化和诱导步骤后可见一蛋白特异高效表达 .Western印迹结果证明 :该蛋白能与兔抗人CK2α′3 3 3 3 50 肽段抗血清发生特异性免疫反应 .采用GSH Sepharose 4B柱纯化 ,凝血酶酶切 ,最后从 4g细菌获 4 4mg纯化重组蛋白 .通过性质鉴定和酶动力学分析证明 :克隆、表达和纯化的重组蛋白是有生物学活性的人CK2α′亚基 .     

CCR7和B7-2在抗原负载树突状细胞诱导特异性CTL抗肿瘤效应中的表达和意义

目的:研究CCR7(趋化因子受体7)和B7-2(白细胞分化抗原86)与抗原负载树突状细胞(dentritic cell,DC)诱导特异性CTL(细胞毒性T淋巴细胞)抗肿瘤效应的关系.方法:分离和培养DC,制备B16黑色素瘤细胞抗原,进行共培养,即为抗原负载的DC,建立B16黑色素瘤小鼠模型,于肿瘤周围皮下注射抗原负载的DC.应用原位杂交和免疫组织化学方法检测CCR7和B7-2的表达情况.结果:原位杂交和免疫组织化学染色显示,CCR7和B7-2阳性细胞主要分布于肿瘤周围组织,随着注射抗原负载DC时间的进展,CCR7和B7-2呈强阳性表达.结论:CCR7和B7-2的表达与抗原负载树突状细胞诱导特异性CTL抗肿瘤效应有关.     

B7家族的新成员

T细胞的活化需要共刺激分子和其受体及特异性抗原与T细胞受体的双信号作用。B7家族成员是重要的共刺激分子,除B7－1和B7－2,新近又发现了一些新成员：B7RP－1（又名B7h,GL50或ICOSL）为鼠ICOS（inducible costimulator,CD28家族的第三位成员）的配体,其人的同源物命名为B7－2（也称为GL50或ICOSL）,它对TG细胞生长及细胞因子产生的共刺激作用已明显,B7－H1（也称PD－1L）,B7H3是一类不与ICOS结合的B7家族新成员。现已证实。现已证实,这些分子与其受体之间的作用在T细胞增殖及发挥效应中扮演重要角色,它们在许多疾病中的作用也引起学者的普遍关注。     

One step engineering of T7-expression strains for protein production: increasing the host-range of the T7-expression system

The T7-expression system has been very useful for protein expression in Escherichia coli. However, it is often desirable to over-express proteins in species other than E. coli. Here, we constructed an inducible broad-host-range T7-expression transposon, which allows simple one-step construction of T7-expression strains in various species, providing the option to over-express proteins of interest in a broader host-range. This transposon contains the T7 RNA polymerase driven by the lacUV5 promoter, which is repressed by the lac-repressor. Leaky expression is prevented by the presence of T7-lysozyme on this construct. The complete T7-expression system is flanked by mariner transposon repeats of the suicidal R6Kgammaori plasmid, pBT20-Deltabla. Stable integration of the whole system is possible by a one-step selection for a Flp-excisable Gm(R)-marker. We showed the engineering of E. coli, Pseudomonas aeruginosa, Erwinia carotovora, Salmonella choleraesuis, Agrobacterium tumefaciens, and Chromobacterium violaceum strains with this construct and demonstrated the expression of the Burkholderia pseudomallei Asd protein in these hosts, by induction with isopropyl-beta-d-thiogalactopyranoside (IPTG).     

高原低氧对大鼠大脑皮质生长休止蛋白7表达的影响

目的 探讨高原低氧对大鼠大脑皮质生长休止蛋白7(Gas7)表达的影响.方法 36只大鼠随机分为正常对照组和模拟高原低氧组,模拟高原低氧组大鼠进行6周缺氧,复制慢性高原低氧动物模型.实验结束后,所有动物采用免疫组织化学和免疫印迹技术检测大鼠大脑皮质中Gas7的表达.结果 与对照组相比,Gas7在模拟高原低氧组大鼠大脑皮质的表达明显增强.结论 Gas7可能参与了高原低氧对大鼠大脑皮质神经元结构和功能的影响.     

BRD7的亚细胞定位及其假定核输出信号序列的分离与鉴

BRD7被鉴定为一个鼻咽癌密切相关新基因和潜在的核转录调节因子．通过绿色荧光蛋白(GFP)介导的亚细胞定位方法,系统研究BRD7在非洲绿猴肾COS7细胞、人宫颈癌HeLa细胞以及人鼻咽癌HNE1细胞中的亚细胞定位,发现BRD7主要定位在细胞核,呈细点状或条梭状分布,3株细胞中没有明显的细胞类型差异．通过对BRD7编码蛋白氨基酸序列进行比对分析,发现了1个具有亮氨酸富集特征的假定核输出信号序列pNES,该区域具有类似核输出信号特征序列“ L-x(2,3)-［LIVFM］-x(2,3)-L-x-［LI］ "（X代表任意氨基酸）的结构;通过功能分析,发现它不具有介导异源蛋白GFP胞浆定位的功能,且其亚细胞定位或胞浆/胞核分布比例不受细霉素B(leptomycin B)干预的影响,说明这个pNES不具核输出信号结构域的功能,不是BRD7的核输出信号．     

Simultaneous determination of coumarin, 7-hydroxycoumarin and 7-hydroxycoumarin glucuronide in human serum and plasma by high-performance liquid chromatography

A HPLC method was developed for the determination of the metabolites of coumarin and 7-hydroxycoumarin in plasma and serum. Separation was based on gradient elution of 7-hydroxycoumarin glucuronide, 7-hydroxycoumarin, coumarin and finally 4-hydroxycoumarin (which is used as an internal standard). Standards, prepared in plasma or serum, and samples were treated with trichloroacetic acid, mixed and centrifuged. The supernatant was removed and analyzed by reversed-phase high-performance liquid chromatography on a C₁₈ column. The limit of detection was 50 ng/ml for 7-hydroxycoumarin and 200 ng/ml for coumarin and 7-hydroxycoumarin glucuronide. The linear range was 0.5–100 μg/ml for each of the analytes. The percentage relative standard deviation about the mean measured concentrations were all below 10%. There was no statistical difference between the standard curves prepared in plasma or serum. The method developed was applied to the determination of each of the three compounds in serum, after the administration of 7-hydroxycoumarin, and in plasma after the administration of coumarin. The concentrations of total 7-hydroxycoumarin in the serum samples were also determined by another HPLC method and the results were compared. There was no statistical difference between the results determined.     

Development of a sensitive liquid chromatography–electrospray ionization tandem mass spectrometry method for the measurement of 7-cyanoquinocarcinol in human plasma

A sensitive method for the determination of an anti-cancer agent, DX-52-1 (7-cyanoquinocarcinol, I) and quinocarmycin (II) which is formed from I either by metabolism or degradation, in human plasma has been developed utilising liquid chromatography electrospray–ionization tandem mass spectrometry (LC–ESI-MS–MS). The procedure involves solid-phase extraction at pH 2 and low temperature (4–6°C) to prevent the decomposition of I to II, the separation by reversed-phase HPLC and the multiple reaction monitoring (MRM) by ESI-MS–MS. The mean precision and accuracy at the lower limit of quantitation (LLOQ) of I, 0.25 ng ml⁻¹, were 8.7% and −10.8%, respectively. Since an interfering peak eluting slightly earlier than II was observed on the HPLC of blank plasma, the LLOQ of II was set at 5 ng ml⁻¹ where the mean precision and accuracy were 15.6% and −9.8%. The results suggested that the method is useful for the simultaneous monitoring of Iand II in the clinical trials of I.