不同级别药用菊花种苗与植株生长及药材产量和品质关系研究

目的:研究不同级别药用菊花种苗与植株生长、药材产量和品质的影响,为药用菊花规范化栽培提供依据。方法:对药用菊花种苗的株高、地径、根长、全株重、地上部分重、地下部分重和高径比7个形态指标进行检测,并进行主成分分析和相关性分析确定分级指标,采用逐步聚类分析方法聚类分级;进行不同等级种苗盆栽试验,对植株生长指标,药材产量和品质指标进行测定。结果:确立以苗高和地径为种苗分级的质量指标,并初步制定了药用菊花种苗质量分级标准;不同级别药用菊花种苗对植株生长、药材外观形态和产量有影响,Ⅰ级苗和Ⅱ级苗的地径和分枝数要多于Ⅲ级苗,而Ⅲ级苗的株高要比Ⅰ级苗和Ⅱ级苗要高。Ⅰ级苗和Ⅱ级苗分枝数多,花心直径和管状花数显著大于Ⅲ级苗,药材产量也显著高于Ⅲ级苗药材产量。而对药材内在品质无明显影响。结论:移栽时种苗以Ⅰ级苗和Ⅱ级苗为佳。     

知母种苗质量分级标准研究

目的:研究制定符合生产实际的知母种苗质量分级标准,为知母药材规范化生产提供依据。方法:对知母主产区基地以及药材市场销售的种苗进行质量指标调查,通过相关分析、主成分分析,并结合生产实践确定知母种苗的质量分级指标,通过K-均值聚类分析将知母种苗分为3个等级,并以不同等级种苗为处理材料进行田间试验,连续2年观测植株生长、药材产量及质量指标。结果:确定种球直径和百株质量作为知母种苗的质量分级指标,将种苗分为3个质量等级,Ⅰ级种苗:种球直径≥1.2 cm,百株质量≤2.0 kg;Ⅱ级种苗:种球直径≥0.8 cm,百株质量≤1.5 kg;Ⅲ级种苗:种球直径≥0.5 cm,百株质量≤1.0 kg且≥0.5 kg。不同等级种苗对植株根系生长、药材产量有显著影响,均表现为:Ⅰ级>Ⅱ级>Ⅲ级。不同等级种苗种植后药材的有效成分含量差异不显著,但均高于《中华人民共和国药典》的标准。结论:本研究制定的知母种苗质量标准即具科学性,也更符合生产实际,可为知母药材的规范化栽培提供理论依据。     

广东不同地区三叉苦营养袋种苗的生长评价和分级标准研究

目的:为三叉苦营养袋种苗质量评价标准的制定提供参考依据。方法:收集广东不同地区三叉苦营养袋种苗,通过测定种苗的地径(D)和株高(H)等形态指标,采用数理统计方法,确定主要分级指标,从而制定种苗分级标准。结果:三叉苦营养袋种苗的分级标准为:Ⅰ级种苗,D≥0.55 cm、H≥40 cm;Ⅱ级种苗,0.55 cmD≥0.45 cm、40 cmH≥30 cm;Ⅲ级种苗,D0.45 cm、H30 cm;平远和五华Ⅱ级以上的种苗占比分别为93.5%和93%,其生长最好,而高州种苗几乎均为Ⅲ级,占比高达92.5%,其素质最差。结论:建立了简单、方便、可行的三叉苦营养袋种苗分级标准,可为三叉苦药材的规范化生产提供一定的参考依据。     

金线莲形态学性状与产量形成关系的多重分析

以13份金线莲种质为研究对象,对其9个性状（株高、地径、叶长、叶宽、植株叶面积、叶片鲜重、叶片数、高径比、植株鲜重）进行相关、通径和主成分分析。不同种源金线莲形态学性状变异系数为2.96%~12.59%,相关性研究表明植株鲜重与地径、株高和叶片数呈极显著正相关,与叶片鲜重呈显著正相关。通径分析表明株高对植株鲜重有最大的正向直接作用,并通过叶片鲜重和叶片数对植株鲜重有不同程度的间接作用。通过主成分分析,将金线莲形态学性状分为“高产型形态决定因子”和“产量的叶部决定因子”。在实际生产及良种选育过程中,应关注金线莲株高、叶片鲜重和叶片数等性状。     

蓬莪术种姜分级及其对植株生长与药材产量和品质的影响

目的 制定蓬莪术种姜分级标准，并探究种姜分级对蓬莪术生长、产量和品质的影响。方法 测定蓬莪术种姜样品的净度、种姜粗、种姜重、种姜长、发芽率、含水量指标，运用偏相关分析、方差分析及聚类分析，确定种姜分级指标，结合生产实际，制定蓬莪术种姜分级标准；然后以不同等级种姜为处理进行田间试验，分析种姜等级对植株生长、药材产量与品质的影响。结果 以种姜粗、种姜重为分级指标，将蓬莪术种姜分为3个等级。田间试验表明从植株生长情况分析，随着蓬莪术植株的生长发育，不同等级种姜植株的株高、叶长在块根膨大期时差异达显著性水平（P<0.05）；从药材产量分析，各级种姜所产郁金、莪术药材产量大小均为一级种姜>二级种姜>三级种姜>等外级种姜；从药材商品等级占比分析，种姜等级越高所产一等郁金药材占比也越高；从药材品质分析，不同等级种姜所产郁金及莪术药材的浸出物、挥发油、吉马酮含量均无显著性差异。结论 通过对蓬莪术种姜分级方法进行系统研究，制定了种姜分级标准；通过田间试验发现，种姜分级种植能促进植株生长发育、增加药材产量、提高一等药材的比例且不影响药材内在品质，可见蓬莪术种姜分级具有科学性和必要性，能为实现蓬莪术的规范化种植奠定基础。     

阳春砂种苗质量分级标准

目的：建立阳春砂种子苗和分株苗的质量分级标准,规范阳春砂种苗生产流通,保障阳春砂规范化、规模化种植。方法：在全国砂仁主产区收集砂仁种子苗和分株苗,测定各种苗的株高、叶片数、丛芽数、球状茎粗、匍匐茎粗等指标,采用主成分分析和K类中心聚类法对数据进行分析并完成砂仁种苗分级。结果：阳春砂种苗可分为2个种类,每个种类2个等级。Ⅰ级种子苗：苗高≥40 cm,丛芽数≥3个;Ⅱ级种子苗：15 cm≤苗高＜40 cm,1个≤丛芽数＜3个;Ⅰ级分株苗：60 cm≤苗高＜110 cm,5片≤叶片数＜10片;Ⅱ级分株苗：110 cm≤苗高＜180 cm,10片≤叶片数＜20片。结论：阳春砂种苗质量分级标准适用于该药材的规范化生产。     

欧李扦插苗质量分级标准研究

目的:制定欧李扦插苗质量分级标准,为郁李仁(欧李)药材规范化生产奠定基础。方法:采集河南省洛阳市新安县一年生欧李扦插苗30份,每份15株,总共450株,测定株高、地径、分枝数、主根长、主根直径、侧根数,利用Excel 2010和SPSS 19.0软件进行统计、分析,确定分级的主要指标,并制定扦插苗分级标准。结果:株高、地径和分枝数是欧李扦插苗质量分级的主要指标。结论:制定欧李扦插苗的3个等级,一级苗:株高≥52 cm,地径≥5.0 mm,分枝数≥2.9;二级苗:52 cm株高≥15 cm,5.0 mm地径≥3.3 mm,2.9分枝数≥1.5;三级苗:株高15 cm,地径3.3 mm,分枝数1.5。     

动态聚类法确定黄连种苗分级标准

目的:制定黄连种苗质量分级标准。方法:测量19个不同产地黄连种苗的株高、叶片数、叶长、叶宽、叶重及根重等指标,通过对各指标数据的相关性统计分析,并结合生产实际,确定黄连种苗质量分级的主要指标和参考指标。采用动态聚类确定黄连种苗的质量分级标准。结果:叶数是黄连种苗质量分级的主要指标,株高作为质量分级的低优指标。结论:黄连种苗分为3级,Ⅰ级种苗:叶数≥8片,14≥株高≥12 cm;Ⅱ级种苗:8≥叶数≥6,12 cm≥株高≥9 cm;Ⅲ级种苗:6≥叶数≥4,株高≥9 cm;达不到3级种苗标准的为不合格种苗。     

枳壳种苗质量分级标准研究

收集重庆、四川、湖南、江西等枳壳主产地43份一年生枳壳嫁接苗,测定株高、砧木粗、地径、根长、根粗、侧根数、根幅、分枝数、分枝长、绿叶数、叶长、叶宽、是否有刺等指标。通过SPSS 19.0软件进行K-均值聚类法分析获得分级标准,结合生产实际,将株高、地径、分枝数作为枳壳种苗(一年生嫁接苗)质量分级指标。制定3个等级分级标准,一级苗:株高不小于80 cm,地径不小于9.0 mm,分枝数不少于5枝;二级苗:株高60～80 cm,地径8.0～9.0 mm,分枝数3～5枝;三级苗:株高不小于40 cm,地径不小于6.0 mm,分枝数不少于1枝。不满足三级标准则为不符合规定苗,不能作为种苗使用,生产上建议使用一、二级苗。     

玄参子芽分级标准研究

对全国玄参主产区子芽的长、粗和重3个形态指标进行检测,使用SPSS统计软件,通过聚类分析制定玄参子芽的质量分级标准。田间试验观测不同等级子芽玄参的植株生长情况及产量。田间试验表明,Ⅰ级玄参长势最好,产量最高,Ⅲ级玄参长势最差,产量最低。玄参药材中主要活性成分含量测定结果表明不同等级的子芽对药材的内在品质无明显影响。为保证高产及减少成本,玄参生产用子芽应控制在Ⅱ级以上。此方法制定的质量分级标准科学、可行,可作为玄参子芽的质量控制标准,为玄参规范化栽培提供依据。     

Investigation of plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts

Aim of the study

To investigate the activities of the 217 plant extracts in traditional medicine of the Brazilian Cerrado against protozoans and yeasts.

Materials and methods

Plant extracts were prepared by the method of maceration using solvents of different polarities. The growth inhibition of chloroquine-resistant Plasmodium falciparum strain (FcB1) was determined by measuring the radioactivity of the tritiated hypoxanthine incorporated. Activity against Leishmania (Leishmania) chagasi and Trypanosoma cruzi was measured by the MTT colorimetric assay. The antifungal tests were carried out by using the CLSI method. The active extracts were tested also by cytotoxicity assay using NIH-3T3 cells of mammalian fibroblasts.

Results

Two hundred and seventeen extracts of plants were tested against Plasmodium falciparum. The eleven active extracts, belonging to eight plant species were evaluated against L. (L.) chagasi, Trypanosoma cruzi, yeasts and in NIH-3T3 cells. The results found in these biological models are consistent with the ethnopharmacological data of these plants. The ethyl acetate extract of Diospyros hispida root showed IC₅₀ values of 1 μg/mL against Plasmodium falciparum. This extract demonstrated no toxicity against mammalian cells, resulting in a significant selectivity index (SI) of 435.8. The dichloromethane extract of Calophyllum brasiliense root wood was active against Cryptococcus gattii LMGO 01 with MIC of 1.95 μg/mL; and Candida albicans ATCC 10231 and Candida krusei LMGO 174, both with MIC of 7.81 μg/mL. The same extract was also active against Plasmodium falciparum and L. (L.) chagasi with IC₅₀ of 6.7 and 27.6 μg/mL respectively. The ethyl acetate extract of Spiranthera odoratissima leaves was active against Cryptococcus gattii LMGO 01 with MIC of 31.25 μg/mL, and against Plasmodium falciparum with IC₅₀ of 9.2 μg/mL and Trypanosoma cruzi with IC₅₀ of 56.3 μg/mL.

Conclusion

The active extracts for protozoans and human pathogenic yeasts are considered promising to continue the search for the identification and development of leading compounds.     

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs

Ethnopharmacological relevance

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims.

Aim of the study

The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs.

Materials and methods

Thirty-six Dorper × Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5 mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks.

Results

Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P > 0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment.

Conclusions

Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

中国石斛属植物文献计量研究

石斛是珍稀濒危中药材,目前正处于快速发展阶段。为全面了解我国石斛属植物研究的历史和发展现状,作者以1954～2010年"中国知网中国学术期刊网络出版总库"收录的石斛研究文献为依据,采用文献计量学的原理和方法,对我国石斛属植物研究文献从文献年代分布、期刊分布与被引频率、主题分布、研究对象分布、作者与研究机构分布等方面进行了统计与分析。结果表明,我国石斛研究明显分为起步(2个)、停滞、平稳发展、快速上升5个阶段;期刊分布存在离散性与集中性并存的现象,已形成核心期刊研究群,并以《中国中药杂志》、《中草药》和《陕西中医》为代表;研究主题广泛涉及临床与药理、组织培养与种苗繁育、成分分析等多个领域,已经形成比较稳定的研究机构和团队,但研究对象差异显著,以铁皮石斛、金钗石斛和霍山石斛最为集中。我国石斛属植物的研究已取得显著成果,但种植产业发展缓慢,供需矛盾突出,预计种苗繁育与人工种植、产品开发、化学与药理等方面是未来的研究热点,其文献报道仍将进一步上升。     

Antifungal activity of Brazilian medicinal plants involved in popular treatment of mycoses

A survey of medicinal plants used to treat common mycoses was done in the Curituba district, Sergipe State, Brazil. One hundred inhabitants were interviewed by health agents and traditional healers. Four different plants were the most cited (more than 50% of the citations): Ziziphus joazeiro, Caesalpinia pyramidalis, Bumelia sartorum and Hymenea courbaril. The aqueous extracts obtained following traditional methods and using different parts of these plants, were submitted to drop agar diffusion tests for primary antimicrobial screening. Only the water infusion extract of Ziziphus joazeiro and Caesalpinea pyramidalis presented a significant antifungal activity against Trichophyton rubrum, Candida guilliermondii, Candida albicans, Cryptococcus neoformans and Fonsecaea pedrosoi, when compared to the antifungal agent amphotericin B. The minimal inhibitory concentration (MIC) of the bioactive extracts was evaluated by the microdilution method. Best activity with a MIC of 6.5 microg/ml for both extracts was observed against Trichophyton rubrum and Candida guilliermondii. Ziziphus joazeiro and Caesalpinea pyramidalis extracts presented also low acute toxicity in murine models. The present study validates the folk use of these plant extracts and indicates that they can be effective potential candidates for the development of new strategies to treat fungal infections.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Regulation effects on abnormal glucose and lipid metabolism of TZQ-F,a new kind of Traditional Chinese Medicine

Aim of the study

Based on the recipe of the traditional anti-diabetic formula TZQ, we developed TZQ-F, a new formula including 8 fractions isolated from Red Paeony root, Mulberry leaf, Lotus leaf, Danshen root and Hawthorn leaf with a good quality assurance. The study was aimed at fraction preparation and effects of the fractions on abnormal glucose and lipid metabolism.

Materials and methods

The active fractions were obtained by macroporous resin, ion-exchange resin and polyamide resin column chromatographies. HPLC analyses were used for quality control. In vitro mechanism study included DPPH radical scavenging, AGEs formation inhibition, α-glucosidase inhibition and lipase inhibition, and rats on high-fat diet were used for in vivo study.

Results

In vitro mechanism study showed that among the 8 fractions, three of them had inhibition effects on intestinal disaccharase, three with inhibition effects on lipase, and five with effects of free radical scavenging. In vivo study showed that after 4 weeks of treatment, TZQ-F significantly decreased the levels of serum total cholesterol, TG, glucose, LDL-C and HDL-C in rats on high-fat diet. Consistent with the in vitro and in vivo results, histology study demonstrated that TZQ-F alleviated hepatic steatosis induced by high-fat diet.

Conclusions

TZQ-F possesses the potential regulation effects on abnormal glucose and lipid metabolism.     

常用藏药“蒂达”(藏茵陈)基原物种药用合理性及资源利用价值评价

目的:对“蒂达”各品种不同基原植物药用合理性及其资源利用价值作出评价.方法:采用HPLC指纹图谱共有特征峰、结合SPSS聚类分析方法,评价“蒂达”不同品种基原植物的成分组成相似性.结果:“蒂达”不同品种涉及的来源有龙胆科獐牙菜属、花锚属、扁蕾属及肋柱花属共15种1变种33个样品,指纹图谱全谱相似度较差;以10个共有峰成分的相对面积聚类,其基原植物可分别聚为4个组;上述4个属的属间成分组成差异较大;其中獐牙菜属不同种间成分组成变化较大,而其他各属种间成分组成相似度较高.结论:上述龙胆科4属植物均含有具有保肝利胆生物活性的环烯醚萜类、(山)酮类及三萜类成分,藏医学将其归为“蒂达”一类药材使用具有一定的物质基础;鉴于“蒂达”类药材不同基原各品种间在成分组成上差异较大,分别制定各品种的HPLC指纹图谱,是鉴别和控制各品种质量的有效手段,可有效控制各基原品种质量.     

虫草属EST-SSR标记系统的建立研究

目的:通过球孢虫草、蛹虫草EST设计EST-SSR引物,建立虫草属EST-SSR标记系统.方法:从NCBI公共数据库下载获得虫草EST,利用Sequece Seiners 1.2软件去除冗余序列并设计引物,进行PAGE电泳.结果:通过去除EST总序列中低质量的和冗余的序列后,得到全长为2 953 173 bp的4 556条无冗余球孢虫草EST.从中发掘出718个EST-SSR,分布于616条EST中,出现频率是15.8％.平均分布频率是每4 096 bp出现1个,三核苷酸重复序列有419个,是出现最多的重复类型.蛹虫草EST去冗余后得到1 363条无冗余EST,共含有1 117个EST-SSR,出现频率为81.95％,出现最多的重复类型是A核苷酸重复.根据球孢虫草EST-SSR序列,设计合成50对引物,有扩增产物的引物为34对,占总设计引物数的68％.根据蛹虫草EST-SSR,设计合成40对引物,有扩增产物的引物为39对,占总设计引物数的97.5％.基于SSR标记进行聚类分析,7种虫草无性型均能分开,且分为4支.结论:虫草属EST-SSR出现频率较高、类型较丰富、多态性潜能较高,具有较高的利用价值.球孢虫草和蛹虫草EST开发的SSR标记在虫草属有较好的转移性与通用性,可以很好的应用于虫草种间遗传关系的研究.应用虫草物种EST建立分子标记是一条简便而又有效的途径.     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。