New Macrocyclic Diamide from Rauvolfia Yunnanensis Tsiang

A new macrocyclic diamide, 22-membered macrocyclic diamide, named cyclodicaprylamide（2）, and five known compounds, bis（2-ethylhexyl） phthalate（1）, ethyl 3,4,5-trimethoxybenzoate（3）, ethyl 3,4,5-trimethoxycinnamate （4）, （＋）-syringaresinol（5）, loliolide（6）, were isolated from the roots of Rauvolfia yunnanensis Tsiang. Their structures were elucidated based on NMR, 2D NMR, and MS spectrum, respectively. They were obtained from it for the first time.

A new monoterpenoid indole alkaloid from Rauvolfia yunnanensis

One new monoterpenoid indole alkaloid, l l-methoxyburnamine-17-O-3′,4′,5′-trimethoxybenzoate （1）, was isolated from Rauvolfia yunnanensis Tsiang. Its structure was identified by speclroscopic evidences.

Mosquito larvicidal activity of Rauvolfia serpentina L.seeds against Culex quinquefasciatus Say

Objective:To establish the larvicidal activities,if any of solvent extracts of Rauvolfia serpentina (R.serpentina) L.seeds against Culex quinquefasciatus(Cx.quinquefasciatus) Say.1823 as target species.Methods:Seeds of R.serpentina were extracted with five solvents graded according to the polarity[viz.petroleum ether.benzene,ethyl acetate,acetone and absolute alcohol] continuing one after another with the same seeds.Results:Mortality rate with petroleum ether extract was significantly higher than other extracts.The mortality rates of late 3rd instar larvae wore 50.33±5.51.10.00±1.00.0.00±0.00.21.33±1.53 and 0.00±0.00 in 100 ppm concentration of petroleum ether,benzene,ethyl acetate,acetone and absolute alcohol respectively,after 24 h of exposure period.Conclusions:Results of this study show that petroleum ether extract of R. serpentina seed may be considered as a potent source of mosquito larvicidal agent.

Evaluation of in-vitro antibacterial activity and anti-inflammatory activity for different extracts of Rauvolfia tetraphylla L.root bark

Objective:To assess the in-vitro antihacterial activity and anti-inflammatory activity of orally administered different extracts(Hydro-alcoholic,methanolic,ethyl acetate and hexane)of Rauvolfia tetraphylla(R.tetraphylla)root bark in Carrageetiaii induced acute inflammation in rats.Methods:In-vitro antibacterial activity was evaluated for extracts against four Gram positive and four Gram negative bacteria by using cylinder plate assay.Hydro-alcoholic extract(70%v/v ethanol)at 200,400 and 800 mg/kg doses and methanolic,ethyl acetate and hexane extracts at doses 100,200 and 400 mg/kg were tested for anti-inflammatory activity in Carrageenan induced rat paw oedema model and paw thickness was measured every one hour up to 6 hrs.Results:All extracts of R.tetraphylla root bark showed good zone of inhibition against tested bacterial strains.In Carrageenan induced inflammation model,hydro-alcoholic and methanolic extract of R.tetraphylla root bark at three different doses produced significant(P<0.00l)reduction when compared to vehicle treated control group and hexane,ethyl acetate extracts.Conclusions:In the present study extracts of R.tetraphylla root bark shows good in-vitro antibacterial activity and in-vivo anti-inflammatory activity in rats.

Cloning and Characterization of the Strictosidine-β-D-glucosidase (SGD) Gene from Rauvolfia verticillata

[Objective] This study aimed to clone Scrictosidine-β-D-glucosidase (SGD) gene from Rauvolfia verticillata and analyze its characteristics. [Method] The full-length cDNA of SGD was cloned from R. verticillata with RACE technique. Then the expression levels in different tissues were analyzed with quantitative RT-PCR and the bioinformatic characteristics were also predicted. [Result] The full-length cDNA of RvSGD was 1 856 bp, containing a 1 608 bp CDS that encoded 536 amino acids with a calculated molecular mass of 61.0 kDa and an isoelectric point of 6.16. Bioinformatic analysis revealed that RvSGD shared high similarity with SGDs from Cantharanthus roseus and Rauvolfia serpentina at the amino acids; three conserved catalytic sites His-161, Glu-207 and Glu-419 were also presented in RvSGD. Quantitative RT-PCR showed that expression level of RvSGD was the highest in barks, followed by old leaves, roots, tender leaves and tender stems. [Conclusion] The present study helps to understand more about the functions of the SGD gene at the level of molecular genetics, and provides new targets for molecular regulation of TIAs biosynthesis.

Rauvotetraphyllines A-E, new indole alkaloids from Rauvolfia tetraphylla

Five new indole alkaloids rauvotetraphyllines A-E(1-5),together with eight known analogues,were isolated from the aerial parts of Rauvolfia tetraphylla.The structures were established by means of spectroscopic methods.

Hybrid Monoterpenoid Indole Alkaloids Obtained as Artifacts from Rauvolfia tetraphylla

Five new hybrid monoterpenoid indole alkaloids bearing an unusual 2,2-dimethyl-4-oxopiperidin-6-yl moiety,namely rauvotetraphyllines F–H(1,3,4),17-epi-rauvotetraphylline F(2)and 21-epi-rauvotetraphylline H(5),were isolated from the aerial parts of Rauvolfia tetraphylla.Their structures were established by extensive spectroscopic analysis.The new alkaloids were evaluated for their cytotoxicity in vitro against five human cancer cell lines.

Three new indole alkaloids from Rauvolfia yunnanensis

One rare tetracyclic macroline-type indole alkaloid,named rauvoyunine A(1),and two new picraline-type alkaloids rauvoyunines B and C(2 and 3)were isolated from the aerial parts of Rauvolfia yunnanensis.Their structures were elucidated on the basis of extensive spectroscopic analysis.Compounds 2 and 3 were evaluated for their in vitro cytotoxicity against five human tumor cell lines.

Extractive from Rauvolfia verticillata (Lour.) Baill. var. hainanensis Tsiang ameliorate ulcerative colitis in Dextran Sodium Sulfate-induced Mice via regulating the MAPK pathway

Objective: To investigate the effect and mechanism of Rauwolfia verticillata (Lour.) Baill. var. hainanensis Tsiang extract on mice with colitis induced by Dextran sulphate sodium (DSS). Methods: 80 BALB/C mice were randomly divided into (1) normal control group, (2) DSS group, (3) DSS + SASP treatment group, (4) DSS + PP treatment group. After the experiment, the mice in each group were observed daily general conditions, Immunohistochemistry was used to detect the expression of ERK, p38 MAPK and JNK protein in colonic mucosa of mice in each group. The expression of TNF-_and IL-6 cytokines in colonic tissue of mice was detected by ELASA. Results: The general situation of SASP group and PP group was better than DSS group;Compared with DSS group, the expression of ERK, p38 MAPK and JNK protein was significantly decreased. The levels of TNF-a and IL-6 in colon tissue of SASP group and PP group were significantly decreased. Conclusion: The results suggest that pectin polysaccharide from Rauvolfia hainanensis extract has a significant therapeutic effect on colitis in DSS mice, which may via regulating the MAPK signal pathway pathways.

Rauvolfia serpentina-Mediated Green Synthesis of CuO Nanoparticles and Its Multidisciplinary Studies

Copper oxide nanoparticles （CuO Nps） were successfully synthesized by solution combustion method using aqueous leaf extract of Rauvolfia serpentina as a fuel. The structure and morphology of the CuO nanoparticles （Nps） were characterized by powder X-ray diffraction （PXRD）, UV-visible spectroscopy （UV-visible）, scanning electron microscopy （SEM）, transmission electron microscopy （TEM）, etc. The PXRD patterns reveal the formation of monoclinic phase with crystallite structure. SEM images indicate that the particles have sponge-like structure being highly porous and agglom- erated with large surface area. The average crystallite sizes were found to be in the range of 10-20 nm by Scherrer＇s method. The CuO Nps size was further confirmed by TEM. Further, CuO Nps exhibit good photocatalytic activity for the photodegradation of trypan blue dye, indicating that it acts as a promising semiconducting material. The antibacterial properties of CuO nanoparticles were investigated against pathogenic bacterial strains, namely Gram -ve Escherichia coli （NCIM-5051） and Pseudomonas desmolyticum （NCIM-2028） and Gram ＋ve bacteria Staphylococcus aureus （NCIM- 5022） using the agar well diffusion method.

HPLC法测定新乐康片相关药材中8种活性成分含量

建立HPLC法同时测定新乐康片中三味原料药材钩藤、酸枣仁和萝芙木中钩藤碱、异钩藤碱、斯皮诺素、酸枣仁皂苷A、酸枣仁皂苷B、育亨宾、阿义吗啉及利血平等8种活性成分的含量。采用Titank C18(250 mm×4.6mm,5μm)色谱柱,以乙腈-0.4%磷酸水溶液(含0.05%三乙胺)为流动相,梯度洗脱,检测波长204 nm,流速为1.0 mL/min,柱温为30℃。上述三味药材相关成分之间分离度良好,在λ=204 nm条件下,其在分别的线性范围内与响应值之间的线性关系良好(R^(2)≥0.9990);8种活性成分的平均加样回收率98.37%~101.21%,RSD<2%,仪器精密度、供试品溶液的稳定性与方法重复性均良好。建立的HPLC法可一法对钩藤、酸枣仁和萝芙木三味中药材中8种活性成分含量进行测定,为药业在生产新乐康片前期选材上节约了测定时间,提高了生产效率,同时为新乐康片中活性成分HPLC含量测定方法的建立及其质量标准升级提供参考。

绿春萝芙木资源薄层色谱鉴别

根据形态学特征对绿春县萝芙木(Rauvolfia L.)资源进行鉴定,同时采用薄层色谱法进行色谱鉴别.结果表明,绿春产萝芙木分别为萝芙木Rauvolfia vericillata(Lour.)Bail.和云南萝芙木Rauvolfia yunnanensisTsiang,两者色谱特征差异明显,但两者均含有药用成份利血平(reserpine)和育亨宾(yohimbine),开发时应综合利用.

云南产萝芙木生物碱薄层色谱法鉴别

采用薄层色谱法对云南产萝芙木[Rauvolfia vericillata(Lour.)Bail.]和云南萝芙木[Rauvolfia yunnanensis Tsiang]生物碱进行色谱鉴别。结果表明,两种萝芙木的根、茎、叶均含有药用成分利血平和育亨宾。鉴于萝芙木各营养器官均含有药用生物碱,开发时应综合利用。

转化毛状根获得萝芙木生物碱的研究

药用植物萝芙木(Rauvolfia verticillata)的无菌苗被含有Ri质粒的发根农杆菌(Agrobacterium rhizogenes)感染后,诱导出毛状根(Hairy root)。将毛状根分离、除菌后,在不含激素的MS固体或液体培养基上培养,从296个株系中筛选出RV 19、RV 26、RV 37和RV53四个生长速度快、分枝数量多等优良特性的无性系。对毛状根中次生代谢产物的提取和测定表明,四个无性系均含有原植物体所具有的吲哚生物碱。为应用转化毛状根技术获得植物次生代谢产物开辟了一条新的有效途径。

高效液相色谱法测定萝芙木中育亨宾和利血平的含量

目的建立检测萝芙木中有亨宾和利血平含量的高效液相色谱分析方法。方法采用C18色谱柱，流动相A为甲醇-水-三氟乙酸（30：70：0．1），流动相B为甲醇-水-三氟乙酸（55：45：0．1），阶段洗脱，流速为1mL／min，检测波长为230nm结果育亨宾、利血平在10～200μg／mL浓度范围内呈良好的线性关系，加样回收率、精密度、稳定性、重复性良好。结论此法灵敏准确，可用于萝芙木中育亨宾、利血平的含量测定。

发根农杆菌介导云南萝芙木的遗传转化

用发根农杆菌A4分别感染云南萝芙木幼嫩的真叶和茎段,诱导出毛状根,并对毛状根进行了离体培养, PCR检测转基因毛状根和HPLC检测毛状根中利血平的含量．结果表明:这两种外植体的转化频率及诱导出的毛状根差异极大,幼嫩的真叶叶片比茎段更适合作转化外植体用．PCR检测结果表明,诱导毛状根的基因rolB和 rolC整合到云南萝芙木基因组中,HPLC含量检测表明毛状根中利血平的含量比自然根中利血平的含量有显著的增加,因此可以作为利血平来源生产的有效途径之一．

萝芙木属植物的物种和分布

对萝芙木属植物的物种及其分布进行了初步研究,目的在于弄清国际上的分类命名现状.结果表明:目前萝芙木属植物的分类比较混乱,还没有一个为国际所普遍接受的分类方法和命名系统,一些种类的分类地位和命名在国际上没有被认可.认为本属植物分类中世界公认的物种应予认可,一些国际权威机构认可的物种可以基本上认定.花和果实是这些植物最重要的分类特征.

IBA和GA_3与温水浸种对催吐萝芙木种子发芽的影响

采用L9（34）和L8（27）的正交设计,了解IBA和GA3与温水浸种对催吐萝芙木种子发芽的影响。结果表明,催吐萝芙木种子平均发芽率和发芽势分别为0.0%-37.7%和0.0%-23.7%,处理组合间呈现极显著的差异。IBA和GA3是影响催吐萝芙木发芽率和发芽势的主导因子,低浓度的IBA（0.25 g/L）和高浓度的GA3（1.00 g/L）与40-60℃的温水配合依次浸种,可极显著提高发芽率和发芽势。

加速溶剂萃取(ASE)技术提取海南萝芙木活性成分

考察了加速溶剂萃取(ASE)技术对海南萝芙木抗氧化活性成分的提取效果。以提取物对DPPH自由基的清除率为指标,确定水-丙酮混合溶剂为最佳溶媒,通过正交实验确定了最佳提取条件,即样品经质量分数1%的盐酸预处理30 m in后,以V(丙酮)∶V(水)=1∶1为溶媒,在125℃用加速溶剂萃取仪提取,所得提取物对DPPH自由基的清除率为89.378%,而索式提取法仅为68.45%。表明加速溶剂萃取技术可以作为萝芙木中活性成分的一种高效提取手段。

ICP-MS测定中药萝芙木中微量元素的含量及形态分析

采用电感耦合等离子体质谱法（ICP-MS）测定了中药萝芙木中9种微量元素的含量及形态分析.样品采用去离子水浸提,HNO3-H2O2电炉加热消解,电感耦合等离子体质谱法（ICP-MS）测定,用茶叶标准物质对测定方法的准确度进行验证,初步探究微量元素的存在形态.结果显示萝芙木中9种微量元素的总含量大小为Zn〉Mn〉Pb〉Fe〉Cu〉Cr〉As〉Ni〉Cd,水提液中各元素的含量大小为Zn〉Mn〉As〉Ni〉Cr〉Pb〉Cd,Fe和Cu未检出.水提残渣态中各元素的含量为0.014-9.643μg/g,水提液中Zn、Mn、Cr、As、Ni元素都存在可溶有机态和可溶无机态,有机态与无机态的比率值在28.12%-264.55%之间,其中Zn、Cr、Ni的有机态大于无机态,比值大于166.67%;颗粒吸附率在7.69%-125%之间.萝芙木中9种微量元素是以多种形态共存的复杂体系.