Chemical composition and larvicidal activity of leaf essential oil from Clausena anisata (Willd.) Hook.f.ex Benth (Rutaceae) against three mosquito species

Objective:To determine the mosquito larvicidal activity of leaf essential oil and their chemical constituents from Clausena anisata（C.anisata）（Willd.） Hook.f.ex Benth.against Culex quinquefasciatus,Aedes aegypti and Anopheles Stephensi.Methods:Essential oil was obtained by hydro-distillation and the chemical composition of the leaf essential oil was analyzed using gas chromatography—mass spectrometry.The mosquitoes were reared in the vector control laboratory and twenty lateⅢinstar larvae of three mosquito species were exposed to based on the wide range and narrow range tests,essential oil was tested at 50.100.150,200 and 250 ppm and each compound was tested at various concentration（5-75 ppm） and were assayed in the laboratory by using the protocol of WHO 2005:the 24 h LC50 values of the C.anisata leaf essential oil and their major compounds were determined following Probit analysis.Results:The oil contained were mainlyβ-pinene（32.8%）,sabinene（28.3%）,germacrene-D（12.7%）,estragole （6.4%） and linalool（5.9%）.The essential oil from the leaves of C.anisata exhibited significant larvicidal activity,with 24 h LC50 values of 140.96,130.19 and 119.59 ppm.respectively.The five pure constituents extracted from the C.anisata leaf essential oil were also tested individually against three mosquito larvae.The LC50 values ofβ-pinene,sabinene,germacrene-D,estragole and linalool appeared to be most effective against Anopheles stephensi(LC50-23.17,19.67,16.95. 11.01,35.17 ppm) followed by Aedes aegypti(LC50-27.69,21.20,18.76,12.70,38.64 ppm) and Culex quinquefasciatus(LC50-32.23,25.01.21.28,14.01,42.28).Conclusions:The essential oil of C. anisata contains five major compounds and has remarkable larvicidal properties,which may be considered as a potent source for the production of natural larvicides.

Contribution of molecular cladistics to the taxonomy of Rutaceae in China

The treatment of Rutaceae in the Chinese flora chose to follow Engler in recognizing Rutoideae and Toddalioideae as two separate subfamilies. Morphological and chemical comparisons, however, suggested grouping those two subfamilies in one subfamily, Rutoideae. This move has received support from molecular phylogenetic analyses, which also showed that the Chinese taxa in Euodia should be placed in Tetradium and Melicope following Hartley. Investigations into the chemistry and molecular phylogeny of Murraya also indicated that the species in the section Bergera without yuehchukene should be removed from Murraya. These findings clearly show the value of molecular cladistics to the taxonomy of Rutaceae in China and also directions for further investigations. Key words Bergera, Euodia, Melicope, Murraya, Rutaceae, Rutoideae, Tetradium, Toddalioideae.

Glycosmis longipetala F.J.Mou & D.X.Zhang,a new species of Rutaceae from China

Glycosmis longipetala F. J. Mou ＆ D. X. Zhang is described from Guangxi and Yunnan provinces in southwestern China. The new species is similar to G. cochinchinensis （Lour.） Pierre ex Engl. by its simple leaves, but distinguishable in having long-elliptic or oblanceolate （vs. ovate） leaves, long-ovoid to ellipsoid （vs. ovoid） floral buds, ovaries with many tubercles （vs. smooth） and glabrous （vs. pubescent） stamens. The pollen grains of the new species are 23.9±3.09 （20.8-27.0）×22.0±1.80 （20.4-24.4）μm in size with reticulate exine ornamentation in equatorial area and foveolate in polar area. The chromosome number of the new species is 2n=72.

A new flavone C-glucoside from Glycosmis arborea(Rutaceae)

A new C-glycosyl flavone 7,4＇-dihydroxy-5-methoxyflavone-6-C-β-D-glucopyranoside 1 has been isolated and characterized form ethanolic extract of leaves part of Glycosmis arborea.The structure of compound was verified by means of high field 1D and 2D NMR（DEFT,COSY,DQF-HSQC,HMBC） and HRMS spectral analysis,respectively.

Molecular Phylogeny of the “True Citrus Fruit Trees” Group (Aurantioideae, Rutaceae) as Inferred from Chloroplast DNA Sequence

The genus Citrus L. has a long controversial taxonomy history, and a well-resolved molecular phylogeny of the ＂true citrus fruit trees＂ group in the future will provide new information for advancing breeding techniques and developing better conservation strategies. In the present study, three cpDNA fragments （TrnL-TrnF, PsbH-PetB, and TrnS-TrnG） of 30 genotypes chosen from the six genera of the ＂true citrus fruit trees＂ group were analyzed. A molecular phylogenetic tree of the ＂true citrus fruit trees＂ group ＂~as reconstructed based on plastid DNA sequences. The results confirmed that the ＂true citrus fruit trees＂ group was monophyletic, and thereby the group was divided into genera as previously suggested based on morphological characters. The cpDNA data also suggested that Poncirus might be the first genus separated from the other five genera in the group. The genus Fortunella were of hybrid origin and Citrus might be as its putative paternal parent. The genera Microcitrus, Eremocitrus, and Clymenia were possibly monophyletic and their common ancestor might branch out from Citrus. Furthermore, the phylogenetic relationships within the Citrus genus were discussed.

20份黄皮种质资源果肉游离氨基酸对风味品质和药用价值的影响及其评价

为研究黄皮果实中游离氨基酸的组分特征、科学评价不同黄皮种质游离氨基酸对果实风味和药用价值的影响,利用高效液相色谱法,测定20份黄皮种质果肉的20种常见游离氨基酸,采用味觉活度值、主成分分析和聚类分析等方法对各种质进行综合评价。结果表明:在20份黄皮种质果肉中均含有除胱氨酸和精氨酸以外的18种游离氨基酸,种类丰富,且多数种质间含量差异显著;各种质的总游离氨基酸质量浓度在1309.7~9832.0 mg/kg之间,其中非必需氨基酸含量最高,必需氨基酸含量次之,半必需氨基酸含量最低;药用氨基酸占总游离氨基酸的6.3%~18.6%,以天冬氨酸和谷氨酸为主;根据氨基酸呈味特性的不同,将18种氨基酸分为3大类呈味氨基酸,发现种质M1的甜味氨基酸含量最高,G2的鲜味氨基酸和苦味氨基酸含量最高;仅16份种质中的丙氨酸与少数种质中的天冬酰胺、天冬氨酸的味觉活度值(taste activity values,TAV)>1,对黄皮果实风味有贡献;大多数游离氨基酸呈显著或极显著正相关关系;主成分分析提取到4个主成分,累计贡献率达到89.766%,能较好地反映出黄皮果肉游离氨基酸的综合特征;各黄皮种质的综合得分在–3.18~3.51之间,M1的综合得分最高;聚类分析将20份黄皮种质聚为3大类,其中第Ⅲ类群的X2、Y-1、G2、M1的总游离氨基酸、药用氨基酸、呈味氨基酸含量高,氨基酸营养品质好,风味品质佳。研究结果可为黄皮良种选育和品质评价提供参考依据。

基于中药特色技术传承概述广东芸香科道地药材

基于中药传统特色技术,文章通过现场观摩调研与文献检索,概述在十大广药中占有1/3的芸香科药物的药用历史、产地采收加工、性状鉴别特点。芸香科药物多含挥发油,具有芳香气,大部分具有宽中理气的功效,临床应用广泛。据考证,柚类橘红始自清代,此前使用橘类橘红;广陈皮主产区在新会地区,也称新会陈皮,药用时间至今约有600年;广佛手以肇庆高要地区产量多且质优,“佛手柑”的记载最早见于元朝。化橘红的传统加工方法是将化州柚的外层果皮切成角状,加工成7角的毛橘红药材,称为“毛七爪”;新会陈皮需在新会地区内陈化三年,方可称为“新会陈皮”。化橘红鉴别特点为表面密被柔软毡状细绒毛,有油点,气清香;新会陈皮“鬃眼”明显,对光照视呈半透明状,香气浓郁。广佛手切面的外皮呈黄色,肉呈白色,俗称“金边白肉”。文章基于中药传统理论、技术对广东地区芸香科道地药材进行概述,以期守正创新,使中药的传统特色技术得到传承发展。

黑果黄皮Clausena dunniana Levl.中一个新的咔唑类生物碱(英文)

从黑果黄皮ClusenadunnianaLevl.中分到一个新的咔唑类生物碱 1,并通过谱学手段确定了 1的化学结构为 3

戊糖片球菌协同蜂蜜接合酵母LGL-1混合发酵对黄皮果酒品质的影响

目的探究戊糖片球菌协同蜂蜜结合酵母LGL-1(Zygosacharomyces mellis LGL-1,LGL-1)混合发酵对黄皮果酒品质的影响。方法从米香型白酒酒厂的小曲中筛选乳酸菌;进行形态学、分子生物学鉴定及生长测定;以乳酸菌协同LGL-1进行果酒酿造;采用顶空固相微萃取-气相色谱-质谱法(headspace solid phase microextraction-gas chromatography-mass spectrometry,HS-SPME-GC-MS)和气相色谱法(gas chromatography,GC)对风味物质进行分析。结果所筛菌株BJQ-1、CZZX-2和LJQ-1为戊糖片球菌(Pediococcus pentosaceus);3株菌皆能利用葡萄糖和果糖进行生长且葡萄糖/果糖质量比1:1复配糖耐受性约300 g/L,BJQ-1、LJQ-1乙醇耐受性达14%vol,CZZX-2乙醇耐受性达16%vol,即所筛菌株适合复配协同酵母菌进行果酒酿造;当CZZX-2、BJQ-1和LJQ-1分别协同LGL-1进行黄皮果酒酿造时,与对照组相比,戊糖片球菌CZZX-2的强化发酵提升黄皮果酒品质的效果最佳,果酒中挥发性物质的种类由32种增加至44种,果酒中2-苯乙醇、异丁醇及异戊醇含量与对照组相比,分别降低24.72%、48.89%和51.57%,乙酸乙酯含量显著提升614.70%。结论戊糖片球菌CZZX-2能协同LGL-1进行黄皮果酒酿造,达到增酯降高级醇的目的,提升其品质。本研究可为黄皮果酒的工业化生产提供菌种及一定的理论基础。

儋州本地黄皮核型分析及同源多倍体诱导

海南省儋州市本地黄皮是著名的热带水果,但存在核大、可食率低、品质不一等问题,为选育无(少)核黄皮新品种,开展儋州本地黄皮核型分析和多倍体诱导试验,对供试黄皮露白种子分别采用0.25%、0.5%、1%秋水仙素和浸泡24、36、48 h 2个因素3个水平共9个处理诱导多倍体,取幼苗根尖进行染色体计数,剥取叶片下表面观测气孔大小和密度,取幼嫩叶片采用流氏细胞仪鉴定倍性,筛选较优的秋水仙素和时间处理后,添加0.05%、0.10%、0.20%二甲基亚砜(DMSO)和5、10、15 mg/L萘乙酸(NAA)优化多倍体诱导方式。结果表明,儋州本地黄皮体细胞染色体数量2n=18,核型公式是2n=2x=18=12m+6sm,相对长度组成是2n=1s+4M1+4M2,2B型,核型不对称性高,进化程度高。0.50%秋水仙碱浸泡36 h处理的下胚轴膨大率最高,为35.53%,多倍体和同源四倍体诱导率最高,分别为4.82%和2.07%,表型变异株中多倍体和同源四倍体占比分别为13.62%和5.83%,为多倍体诱导较优处理。该处理优化以0.10%DMSO+10 mg/L NAA处理最优,多倍体和同源四倍体诱导率最高,分别为18.18%和8.74%,除了0.20%DMSO+10 mg/L NAA外,极显著高于其他处理(p<0.01)。

鸡皮果-百香果复合饮料的研制及抗氧化性研究

为了探究鸡皮果-百香果复合饮料的制作工艺及其抗氧化性,以鸡皮果和百香果为原料,通过单因素和响应面试验,优化复合饮料的制作工艺。结果表明,复合饮料的最佳工艺为百香果汁添加量45%、鸡皮果汁添加量55%、罗汉果苷添加量0.018%、果葡糖浆添加量2%,得到的饮料酸甜适中、风味独特。在此工艺条件下,通过单因素和正交试验,确定最佳稳定剂复配条件为CMC-Na添加量0.08%、黄原胶添加量0.2%;最佳护色剂为茶多酚添加量0.012%、D-异抗坏血酸添加量0.01%复配。评价复合饮料体外抗氧化能力,得到ABTS自由基清除率为96.7%、羟自由基清除率为90.5%。

Two New O-terpenoidal Coumarins, Excavacoumarin A and B from Clausena excavata

Two new O-terpenoidal coumarins named excavacoumarin A (2): B (3), and a known one 1 were isolated from the leaves of Clausena excavata Burm. f. (Rutaceae) collected in Xishuangbanna, Yunnan, Structure elucidation and unambiguous NMR assignments for the title compounds were carried out on the basis of 1D and 2D NMR experiments.

A Novel Dimeric Coumarin from Clausena lenis

A novel dimeric coumarin (1), a dimeric of seselin named diseselin A, was isolated from the aerial part of Clausena lenis. The structure was elucidated based on the MS, 1D and 2D NMR data.

Antiplasmodial and analgesic activities of Clausena anisata

Objective:Antiplasmodial and analgesic activities of the leaf extract and fractions of Clausena anisata(C.anisata) were evaluated for antimalarial and analgesic activities.Methods:The crude leaf extract(39-117mg/kg) and fractions(chloroform and acqeous;78mg/kg) of C. anisata were investigated for antiplasmodial activity against chloroquine-sensitive Plasmodium berghei(P.berghei) infections in mice using suppressive,prophylactic and curative models and analgesic activity against acetic acid,formalin and heat-induced pains.Artesunate,5 mg/kg and pyrimethamine,1.2mg/kg were used as positive controls.Thin films made from tail blood of each mouse were used to assess the level of parasitaemia of the mice.Results:The extract and its fractions dose-dependently reduced parasitaemia induced by chloroquine-sensitive P.berghei in prophylactic,suppressive and curative models in mice.These reductions were statistically significant(P【0.001).They also improved the mean survival time(MST) from 17 to 21 days relative to control(P【0.01-0.001).On chemically and thermally-induced pains,the extract inhibited acetic acid and formalin-induced inflammation as well as hot plate-induced pain in mice.These inhibitions were statistically significant(P【0.001) and in a dose-dependent fashion. Conclusions:The antiplasmodial and analgesic effects of this plant may in part be mediated through its chemical constituents and it can be concluded that the C.anisata possess significant antimalarial and analgesic properties.

Effect of clausenamide on hippocampal neuron apoptosis induced by sodium nitroprusside

BACKGROUND： Aggregation of β-amyloid peptide （A β ）, excitatory intoxication, oxidation injury and inflammation reaction are generally regarded as the main pathogenesis for Alzheimer disease （AD）. （ - ） clausenamide is characterized by promoting intelligent development, resisting oxidation, cleaning free radicals, resisting A β neurotoxicity and nerve cell apoptosis, inhibiting over phosphorylation of tau protein, and improving central cholinergic system. However, whether （-） clausenamide has an effect on hippocampal neuron apoptosis or not need further study. OBJECTIVE： To observe the effect of （ - ） clausenamide on survival rate of hippocampal neurons due to sodium nitroprusside （SNP） and analyze the possible pathways. DESIGN： Contrast observation. SETTING： Institute of Biochemistry and Molecular Biology, Guangdong Medical College. MATERIALS： A total of 12 male SD rats of 24 hours old were provided by the Experimental Animal Center of Guangdong Medical College. The primer was synthesized by Beijing Huada Genetic Engineering Company and （-） clausenamide （99.6%） was provided by Pharmacological Department of Chinese Academy of Medical Sciences. SNP was provided by Sigma Company. METHODS： Bilateral hippocampus was collected from newborn rats to establish single cell suspension. On the 12th day, hippocampal neurons were pretreated with 0.2, 0.4, 0.8 and 1.6 la mol/L （ - ） clausenamide for 6 hours; the culture medium was gotten rid of and neurons were washed with non-serum DMEM solution for three times. In addition, non-serum DMEM solution was added with the above mentioned volume of （ - ） clausenamide and 50 μ mol/L SNP to culture neurons for 24 hours and the collected cells were prepared for the experiment. Neurons were equally divided into control group （culture medium control）, model group （SNP treatment） and experimental group [（ - ） clausenamide ＋ SNP]. Experiment of each group was done for three times at least. Survival rate of cells was measured with MTT chromatometry; levels of mRNA of hippocampal neuron bcl-2 and bax gene were detected with reverse transcription polymerase chain reaction （RT-PCR）; expression of hippocampal neuron Bcl-2 and Bax protein was measured with Western blot technique. MAIN OUTCOME MEASURES： ① Effect of （-） clausenamide on survival rate of SNP-induced hippocampal neuron apoptosis; ②bcl-2 and bax mRNA and protein expression ofhippocampal neurons. RESULTS： ①Survival rate of hippocampal neurons： Survival rate of hippocampal neurons affected by 0.4 - 1.6 μ mol/L （ - ） clausenamide was higher in the experimental group than the model group （P 〈 0.01）, and the survival rate was increased with the larger volume of （ - ） clausenamide. Survival rate was the highest when hippocampal neurons were induced by 1.6 μ mol/L, and it had obvious dosage dependence （P 〈 0.01）. ②Expression of bcl-2 and bax mRNA： Hippocampal neurons were pretreated with 0.2 - 1.6 μ mol/L （ - ） clausenamide for 6 hours in the experimental group and strap of PCR product of bcl-2 gene was brightened gradually. This suggested that, with the increase of concentration, expression of bcl-2 mRNA was increased simultaneously. However, when strap of PCR product of bax gene was darkened, expression of bax was decreased with the increase of concentration. ③Expression of Bcl-2 and Bax protein： Hippocampal neurons were pretreated with 0.2 - 1.6 μ mol/L （ - ） clausenamide for 6 hours in the experimental group and strap of PCR product of Bcl-2 protein was thickened gradually. This suggested that, with the increase of concentration, expression of Bcl-2 protein was increased simultaneously. However, when strap of PCR product of Bax protein was thinned, expression of Bax protein was decreased with the increase of concentration. CONCLUSION： （ - ） clausenamide can resist neurotoxic effect of SNP through dosage dependence, and the mechanism may be related to promoting expression of anti-apoptotic bcl-2 gene and inhibiting expression of pro-apoptotic bax gene.

STUDIES ON THE CHEMICAL CONSTITUENTS OF CLAUSENA LANSIUM(LOUR.)SKEELS Ⅲ.The structural elucidation of homo-and zeta-clausenamide(1)

The structures of two novel lactams isolated from Clausena lansium were elucidated mainly on the bases of their spectral data. They are homoclausenamide(1),a -lactam,and zetaclausenamide(4), an eight-membered ring lactam.

Prenylated Coumarins from Heracleum stenopterum,Peucedanum praeruptorum,Clausena lansium,and Murraya paniculata

Four hitherto unknown prenylated coumarins,namely 600-O-b-D-apiofuranosylapterin(1),40-O-isobutyroylpeguangxienin(2),6-(3-methyl-2-oxobutyroyl)-7-methoxycoumarin(3),and 6-hydroxycoumurrayin(4),were isolated from the ethanol extract of Heracleum stenopterum,Peucedanum praeruptorum,Clausena lansium,and Murraya paniculata,respectively.Their chemical structures were established on the basis of extensive spectroscopic analysis.Compound 2 exhibited in vitro cytotoxic activity against five human cancer cell lines(HL-60,A-549,SMMC-7721,MCF-7,and SW-480)with IC50 values ranging from 15.9 to 23.2μM.

Methanolic extract of Clausena excavata promotes wound healing via antiinflammatory and anti-apoptotic activities

Objective: To investigate the anti-inflammatory properties of methanolic extract of Clausena excavata in lipopolysaccharide(LPS)-activated macrophages(J774 A.1) and the effect on skin wound in a rat model through determining cytokine levels and gene expressions. Methods: The effects of methanolic extract of Clausena excavata on in vitro viability and TNF-α, IL-6, IL-10, and nitric oxide release by LPS-activated J774 A.1 cells were determined. In addition, relative expressions of BAX, BCL-2 and COX-2 genes were examined in healed wounds of rats. Results: The methanolic extract of Clausena excavata was not toxic to J774 A.1 cells at the highest dose of 400 μg/m L. It decreased levels of TNF-α and IL-6, while increasing IL-10 level in LPSactivated J774 A.1 cells and in the healed wounds of rats. The methanolic extract of Clausena excavata also inhibited nitric oxide production in LPS-activated J774 A.1 cells. The BAX and COX-2 genes were downregulated while the BCL-2 gene was upregulated in the healed wound of rats. Conclusions: The methanolic extract of Clausena excavata promotes wound healing via its anti-inflammatory and anti-apoptotic activities.

Clausenlanins A and B,Two Leucine-Rich Cyclic Nonapeptides from Clausena lansium

Two new cyclic nonapeptides,named clausenlanins A(1)and B(2),were isolated from the roots and rhizomes of Clausena lansium.Their structures were elucidated as cyclo-(Gly^(1)-L-Leu^(2)-L-Ile^(3)-L-Leu^(4)-L-Leu^(5)-L-Leu^(6)-L-Leu^(7)-L-Leu^(8)-L-Leu^(9))(1)and cyclo-(Gly^(1)-L-Leu^(2)-L-Val^(3)-L-Leu^(4)-L-Leu^(5)-L-Leu^(6)-L-Leu^(7)-L-Leu^(8)-L-Leu^(9))(2)respectively on the basis of extensive spectroscopic analysis,particularly 2D NMR spectra taken at the temperature of 338 or 303 K and MS.

STUDIES ON THE CHEMICAL CONSTITUENTS OF CLAUSENA LANSIUM (LOUR.) SKEELS Ⅳ THE STRUCTURAL ELUCIDATION OF SECO-AND SECODEMETHYL-CLAUSENAMIDE

The spectral data and chemical properties of two new amides isolated from Clausena lansium led to the assignment of acyclic structures of C-6 and C-7. The absolute configurations of the two chiral carbons in C-6 were determined by degradation to be 3S4R.