Advances in Pharmaceutical and Ecological Value of Sagina japonica(Sw.)Ohwi

Huan WANG, Jingbo HU, Haiyan HUANG, Wenjin XU, Jiao SHANG, Xia ZHANG, Yarong MA, Jiang GONG, Shifeng NI*

1. Department of Computer Science,, Baoji University of Arts and Sciences, Baoji 721007, China;

2. Department of Electronic & Electric Engineering, Baoji University of Arts and Sciences, Baoji 721007, China;

3. College of Life Sciences, Northwest University, Xi’an 710069, China;

4. Department of Medicine, Tibet Nationality College, Xianyang 712082, China

Brief Introduction of Sagina japonica(Sw.)Ohwi

The genus Sagina, belonging to the family Caryophyllaceae, consists of about 30 species, of which four following species are distributed in China:S.japonica(Sw.)Ohwi,S.maxima A. Gray, S. procumbens L., and S. saginoides(L.)Karsten.S.japonica(Sw.) Ohwi, also known as Japanese pearlwort, is widely distributed in Hunan, Hubei, Sichuan, Jiangsu,Guizhou, Yunnan and other regions in China[1].

Identification of S.japonica(Sw.)Ohwi

Morphologic identification

S.japonica (Sw.)Ohwi is an annual or biennial herbelet with clustered stems and opposite leaves, and the bases are connected with films as short sheaths;the flowers are terminal and axillary, white and small; the calyxes are ovate and green with membranous glabrous edges,and separated directly to the bases; there are five stamens; the ovaries are superior,ovoid, with 5 styles that extend outwardly in the shape of a star;there are hair-like projections on stigmas; the capsules are ovoid, about 3 mm in length, which are dehiscent into five valves when they are ripe; the seeds are small, numerous, with projections on the brown testae[2].

S. japonica (Sw.) Ohwi is dry and wrinkled, green or gray green, with one taproot and several thin fibrous roots; the stems are clustered, thin and twisted mostly with vertical ribs,most of which sprout from the bases with obvious nodes and internodes,and are brittle and easily broken[2-3].

Microscopic identification

The upper and lower epidermal cells of S.japonica (Sw.)Ohwi exhibit wavy anticlinal walls. The stomas are densely distributed in a straight axis form. The mesophyll tissues containcalcium oxalate crystals with the diameter of 25-67 μm[2].

Epidermal cells of S. japonica(Sw.) Ohwi are arranged in a column,which are square and covered by stratum corneum. The cortex consists of 4-8 columns of thin-walled cells and contains calcium oxalate crystals.Loop fibers are arranged in 1-2 columns, which are large with thick walls.The phloem consists of 4-6 columns of closely arranged small square or polygonal cells.The xylem vessels are small, arranged in 2-3 circular columns, without xylon. The pith is relatively small and consists of quasi-circular thin-walled cells[3].

Composition Analysis

Zhang and Zhou[4]eluted chloroform extracts of S.japonica(Sw.)Ohwi by silica gel column chromatography and analyzed the elution fraction 4 using gas chromatograph-mass spectrometer; a total of 48 components were identified, mainly including lipids,alkanes and alcohols.

Huang et al.[5]prepared amino acid test solution with acid hydrolysis method to detect the amino acid composition of S. japonica (Sw.) Ohwi and found that S. japonica (Sw.) Ohwi contained a large amount of amino acids, especially for essential amino acids, semi-essential amino acids and drug-effective amino acids.

Zhang and Zhou[6]eluted chloroform extracts of S.japonica(Sw.)Ohwi by silica gel column chromatography and identified 59 components (mainly including alkanes and esters)in elution fractions 1 and 2 using gas chromatograph-mass spectrometer.

Gao et al.[7]reported that rutin and quercetin could be well separated by high-performance liquid chromatography, which exhibited good linear relationships with the peak area within the concentration range of 0.03-15.00 and 0.04 -20.00 μg/ml, respectively;the established method could be used to control the quality of S. japonica(Sw.) Ohwi, thereby providing reference for the development of quality standards of S.japonica(Sw.)Ohwi.

Qin[8]characterized the molecular structures of 28 volatile compounds separated from S.japonica(Sw.)Ohwi with molecular connectivity index and atom-type electrotopological state index, and established a quantitative structure-retention relationship (QSRR)model of the volatile compounds separated from S.japonica(Sw.)Ohwi with the relative standard deviation of 0.997, standard deviation of 0.769,and the calculated values were in good agreement with experimental data.

Li et al.[9]analyzed petroleum ether,water,ethanol and acid extracts of S. japonica (Sw.) Ohwi with test tube method and found that S. japonica (Sw.) Ohwi might contain flavonoids, triterpenoid saponins,amino acids, reducing sugar, organic acids, oil esters, coumarin, inner esters and alkaloids.

Huang and Sun[10]optimized the extraction technology of total flavonoids from S.japonica(Sw.)Ohwi by orthogonal experiments and determined the content of total flavonoids;the optimal extraction conditions were as follows:ethanol concentration 80%,material to liquid ratio 1: 20, extraction time 4 h,extraction temperature 50 ℃.

Zhang and He[11]analyzed the petroleum ether extract of S. japonica(Sw.) Ohwi by gas chromatography/mass spectrometry and identified 25 components, among which aliphatic acids accounted for the highest proportion,and the content of linoleic acid reached 49.318%.

Jia et al.[12]obtained two new phytosterones [22, 25-epoxy-24-methylene-2, 3, 14, 20-tetrahydrocholest-7-en-6-one (japonicone (1))and 20-hydroxyecdysone]from S.japonica(Sw.)Ohwi and investigated the possible biosynthetic pathway of japonicone(1).

Subsequently,Jia et al.[13]isolated and identified two glucocerebrosides[(2S, 3S, 4R, 8E)-1-(β-D-glucopyranosyl-3, 4-dihydroxy-2-[(R)-2′-hydroxypalmitoyl]amino-8-heptadecaene and (2S, 3R, 8E)-1-(β-D-glucopyranosyl-3-hydroxy-2-[(R)-2′ -hydroxypalmitoyl]amino-8-octadecaene) from S. japonica (Sw.) Ohwi with chemical degradation method and spectroscopic method.

Jia et al.[14]isolated two new minor cyclopeptides from S. japonica (Sw.)Ohwi, including aponicin A (1) [cyclo-(Pro1-Pro2-Leu2-Leu1-Phe2-Pro3-Gly-Ser-Phe1)]and japonicin B (2)[cyclo -(Pro1 -Ile -Tyr -Asp -Pro2 -Phe2-Pro3-Phe1].

Jia et al.[15]isolated nine phenolic compounds from S. japonica (Sw.)Ohwi, including p-E-methoxycinnamic acid methyl ester, umbelliferone, 7-methoxy coumarin, 5, 7-di-hydroxy coumarin, 5, 7-di-methoxy coumarin,cerarvensim-7-O-glucoside,5,7,2-trihydroxy-8-methoxy flavone, 5, 7-dihydroxy-8, 2-di-methoxy flavone and 5, 7, 3, 4-tetrahydroxy-6-methoxy flavone.

Huang et al.[16]determined the contents of eight trace elements (K,Na, Ca, Mg, Mn, Cu, Fe and Zn) in S. japonica (Sw.)Ohwi by atomic absorption spectrometry and found that the contents of K,Ca,Mg and Fe were relatively high, followed by Mn, Na, Zn and Cu.

Huang and Li[17]isolated and identified 28 components from the volatile oils of S. japonica (Sw.) Ohwi by gas chromatography/mass spectrometry(GC/MS), including alkanes, amines,acids, alcohols, ketones, aldehydes and phenols.

Pharmacologic Effects

Anticancer effect

Liang et al.[18]treated human leukemia cell line K562 with different concentrations of S. japonica (Sw.)Ohwi extracts and found that the proliferation inhibition rate of K562 cells was enhanced with the increase of extract concentration; S. japonica (Sw.)Ohwi extracts could induce the apoptosis of K562 cells, which might be associated with the downregulation of Bcl-2 expression level, upregulation of Bax expression level and improvement of Caspase-3 and Caspase-9 activities.

Zhang and Zhou[19]investigated the antineoplasmic activity of five different extraction fractions of S. japonica(Sw.)Ohwi on human leukemia cell line K562, human cervical cancer cell line Hela and human breast carcinoma cell line MCF-7 with MTT method and SRB method,and found that S.japonica saponins could effectively inhibit leukemia cells in vitro.

Tian[20]carried out systemic pharmacological experiments on volatile oils, saponins and lavonoid glycosidesisolated from S. japonica (Sw.) Ohwi and found that flavone glycosides had a significant anticancer effect on U16(mouse cervical carcinoma 14)and L615(mouse leukemia 615) in vivo, which was identified as the effective anticancer component of S.japonica(Sw.)Ohwi.

Huang et al.[21]conducted in vivo anticancer experiments and found that flavone glycosides of S.japonica(Sw.)Ohwi had a significant antitumor effect U14; according to the results of acute and subacute toxicity tests, the maximal tolerance dose of dogs to flavone glycosides of S. japonica (Sw.) Ohwi was 0.83 mg/(kg·d).

Others

Huang et al.[22]reported that intraperitoneal injection of 10 mg/kg flavonoid glycosides of S. japonica(Sw.)Ohwi exhibited a significant antiinflammatory effect on rats; flavonoid glycosides of S. japonica (Sw.) Ohwi might also be effective for eliminating local swelling and inflammations.Specifically,at 1-6 h post-injection,local swelling was significantly removed.However,sodium salicylate exhibited a significant anti-inflammatory effect at 2 h post-injection, which was relatively slow compared to flavonoid glycosides.

According to Huang et al.[23],S. japonica (Sw.) Ohwi could stimulate rabbit uterus in vivo and in vitro;small doses could induce rhythmic contractions and large doses could cause tetanic contractions.In addition,S. japonica (Sw.) Ohwi had an inhibitory effect on the hearts of toads and rabbits in vitro because of acting on myocardial M-receptors. S. japonica(Sw.) Ohwi exhibited no inhibitory effect during the immunologic process of rats and rabbits, which was conducive to the application of S.japonica(Sw.)Ohwi as an anticancer medicine.

Huang et al.[24]measured the cough incubation period and cough frequency within four min of mice intraperitoneally injected with 1%S. japonica (Sw.) Ohwi volatile oils and saline under the stimulation of ammonia water and found that S. japonica (Sw.) Ohwi volatile oils had a significant antitussive effect,which could significantly prolong the cough incubation period and reduce cough frequency of mice; however,flavonoid glycosides and saponins of S. japonica (Sw.) Ohwi exhibited no significant antitussive effect.

S.japonica(Sw.)Ohwi had no inhibitory effect during the immunologic process of rats and rabbits but inhibited the hearts of toads and rabbits in vitro that could be resisted by atropine.Therefore,S.japonica(Sw.)Ohwi was harmful to fetal development, and it should be used with caution in pregnant women[23].

Clinical Applications

Pharmacological properties

S.japonica (Sw.)Ohwi is bitter in taste and cool in nature,and endowed with the effects of swelling removing and detoxification[25]. It is commonly used to treat tinea blanca, eczema,erysipelas, scrofula, innominate toxic swelling,snakebite,dental pain and internal traumatic injury[26-27]. Moreover,S.japonica(Sw.)Ohwi is also effective in treating lacquer dermatitis with various functions such as heat clearing,detoxification, swelling removing and itching relieving[28].

Cancer treatment

Xue[29]reported that S. japonica(Sw.)Ohwi had a good effect on treating a patient with acute myelogenous leukemia M2b(AML-M2b).

Nervous system

According to related literature,S. japonica (Sw.) Ohwi could be used to treat toothache:grinding 9 g of fresh S. japonica (Sw.) Ohwi, adding with several drops of vegetable oil and applying the mixture to the affected tooth can effectively relieve the pain[30].

Immune system

According to Xue[29], S. japonica(Sw.)Ohwi was used to treat a patient with acute myelogenous leukemia M2b (AML-M2b), and a good effect was achieved.After radiotherapy,15 g of S.japonica (Sw.)Ohwi and 50 g of semen coicis could be cooked into the porridge for administration to relieve the radiotherapy-induced decrease of white blood cells and anorexia.

Urinary system

Yang reported that decocting 30 g of S.japonica (Sw.) Ohwi for administration could effectively ease urination difficulties[30].

Ecological Value

As a perfect lawn species,S.japonica(Sw.)Ohwi has a large number of excellent features: short plant height,free of pruning, intensive growth,strong soil adaptability, good drought resistance, good treading tolerance,strong restoring ability, easy propagation and convenient planting. S.japonica (Sw.) Ohwi seeds can spread naturally to renew the lawn.Furtherance, S. japonica (Sw.) Ohwi bears white flowers, which can be used to develop ornamental lawns,with broad prospects[28].

Due to wide soil adaptability,S.japonica (Sw.)Ohwi can grow normally in loam, pure forest humus soil, loess and limestone soil, which can grow tall in loose soil and short in compact soil with small plant type. In addition,S. japonica (Sw.)Ohwi can grow well either in an open space or under conditions with canopy density above 80%.It exhibits strong shade tolerance in the vegetative period and poor shade tolerance in the reproductive period. Large scale popularization and cultivation of S. japonica (Sw.) Ohwi can provide abundant resources for the establishment of flowering ornamental lawns. Therefore, wild-type S. japonica (Sw.) Ohwi has a broad prospect as a lawn species[31].

Representive Dietary Prescriptions

Treatment of lacquer dermatitis:mashing S. japonica (Sw.) Ohwi,adding with the juice of luffa leaves,and mixing with vegetable oil for application. Treatment of dental pain:mashing S. japonica (Sw.) Ohwi leaves and stuffing the leaves into the slits between the teeth. Treatment of internal traumatic injury: decocting 25 g of S.japonica(Sw.)Ohwi with water for administration. Treatment of snakebite: mashing S. japonica (Sw.) Ohwi with realgar for application[32]. Treatment of scrofula: decocting 25-50 g of S.japonica (Sw.)Ohwi for administration and mashing fresh S. japonica (Sw.) Ohwi leaves for application[33]. Treatment of abnormal sweating and night sweat:stewing 50 g of S. japonica (Sw.) Ohwi with porkfor eating[34]. Treatment of coughor urination difficulties: cooking 50 g of S. japonica (Sw.)Ohwi in water for administration[35].

Cultivation and Propagation Technologies of S. japonica(Sw.)Ohwi

S. japonica (Sw.) Ohwi is suitable for sowing propagation or transplanting. Before sowing, the ground is leveled,and then S.japonica (Sw.)Ohwi seeds are mixed with fine soil and evenly spread on soil surface, or mixed directly with water and evenly sprayed on the soil[28]. Based on the cultivation test of S. saginoides (L.)Karsten introduced from Tibet, the optimal medium for shoot induction was MS + 6-BA 0.5 mg/L + NAA 0.1 mg/L + sucrose 30 g/L; the optimal medium for rooting was MS + IBA 0.5 mg/L+sucrose 20 mg/L[35].

Conclusions and Prospects

At present,S.japonica(Sw.)Ohwi is widely distributed in China with abundant resources and a long medicinal history, which has been commonly used as a traditional medicine to treat lacquer dermatitis with reliable curative effect. In recent years, a large number of studies have been carried out on the composition,pharmacological effects and clinical applications of S.japonica(Sw.)Ohwi,but little information is available on its anticancer effect. Furthermore,although antitumor constituents of S. japonica (Sw.) Ohwi have been analyzed,patent medicines are not yet developed. Therefore, the medicinal and ecological value of S. japonica(Sw.) Ohwi still needs to be further developed. Moreover, S. japonica(Sw.) Ohwi germplasm resources should be protected to realize sustainable use.

[1]EDITORIAL BOARD OF CHINA HERBAL(中华本草编委会).China Herbal(中华本草) (Volume 2) [M]. Shanghai:Shanghai Scientific and Technical Publishers (上海: 上海科学技术出版社),1999.

[2]ZHANG YF (张云方). Identification of Hedyotis diffusa (白花蛇舌草的真伪鉴别)[J]. China Pharmaceuticals (中国药业),2007,16(8):59-60.

[3]LU BZ (芦柏震), HOU GL (侯桂兰),ZHOU JJ ( 周佳佳). Identification of Hedyotis diffusa from 4 kinds of adulterants(白花蛇舌草与4 种混伪品的辨别)[J]. Chinese Archives of Traditional Chinese Medicine (中医药学刊),2006,24(8):1566-1567.

[4]ZHANG SY (张素英),ZHOU WJ (周万镜).Analysis of chemical constituents of the chloroform extract part from Sagina japonica(Sw.) Ohwi(Ⅱ)(漆姑草氯仿提取物的化学成分分析(Ⅱ))[J].Hubei Agricultural Sciences(湖北农业科学),2013,52(20):5034-5036.

[5]HUANG ZY (黄筑艳),ZHAO YT (赵延涛),ZHANG Z (张振),et al.Analysis on amino acid component from Sagina japonica (漆姑草中氨基酸成分分析)[J].Guizhou Agricultural Sciences (贵州农业科学),2013,41(11):56-57.

[6]ZHANG SY (张素英),ZHOU WJ (周万镜).Analysis of chemical constituents of the chloroform extract part from Sagina japonica(Sw.)Ohwi(漆姑草氯仿提取物的化学成分分析)[J]. Hubei Agricultural Sciences(湖北农业科学),2012,51(15):3326-3329.

[7]GAO ZX(高智席),WU YH(吴艳红),LI G( 黎刚), et al. Contents of rutin and quercetin in Sagina japonica(Sweet)Ohwi determined by HPLC(高效液相色谱法测定漆姑草中芦丁和槲皮素含量)[J].Hubei Agricultural Sciences(湖北农业科学),2012,51(7):1444-1446.

[8]QIN ZL(秦正龙).Quantitative structureretention relationship studies of volatile oil from Sagina japonica (漆姑草挥发油色谱保留值的构效关系研究)[J].Packaging and Food Machinery(包装与食品机械),2012,30(5):9-12.

[9]LI Y (李焱),YAN YH (晏雨寒),HUANG ZY (黄筑艳).Preliminary test of chemical components of Sagina japonica(SW1)Ohwi (中药漆姑草化学成分的初步研究)[J].Guangzhou Chemical Industry(广州化工),2011,39(1):92-93.

[10]HUANG ZY (黄筑艳), SUN Y (孙瑶).Study on the content and the extraction process of the total flavonoids from Sagina japonica(SW1)Ohwi(漆姑草中总黄酮提取工艺研究与含量测定)[J].GuangZhou Chemical Industry and Technology (广州化工),2011,39(19):59-61.

[11]ZHANG SY (张素英), HE L (何林).Analysis of chemical constituents of petroleum ether extract from Sagina japonica (Sw.) Ohwi and screening of its anti-tumor activity (漆姑草石油醚提取物化学成分分析及抗肿瘤活性筛选)[J]. Journal of Anhui Agricultural Sciences(安徽农业科学), 2010, 38(28):15590-15591,16082.

[12]JIA AIQUN,LI YAN,ZHOU JUN,et al.Three phytoecdy steroids from Sagina japonica and potential biotransforming pathways of japonicone [J],Chemistry of Natural Compounds, 2010A, 46(5):738-741.

[13]JIA AQ, YANG X, WANG WX, et al.Glycocerebroside bearing a novel long-chain base from Sagina japonica(Caryophyllaceae)[J].Fitoterapia,2010,81:540-545.

[14]JIA AQ, TAN NH, ZHOU J. Two new minor cyclopeptides from Sagina japonica(Caryophyllaceae)[J]. Fitoterapia,2009,80:192-195.

[15]JIA AQ,(贾爱群), TAN NH,(谭宁华),ZHOU J (周俊). Phenolic compounds from Sagina japonica(漆姑草中酚性成分研究)[J]. Chinese Traditional and Herbal Drugs (中草药), 2008, 39(11):1610-1612.

[16]HUANG ZY (黄筑艳),ZHAO YT (赵延涛),LI Y (李焱),et al. The analysis of trace elements in Sagina japonica (漆姑草中微量元素的分析)[J]. Lishizhen Medicine and Materia Medica Research (时珍国医国药), 2007, 18(5):1131-1132.

[17]HUANG ZY (黄筑艳) , LI Y (李焱). A study on chemical constituents of volatile oil from Sagina japonica (漆姑草挥发油化学成分的研究)[J]. Journal of Changchun Normal University Natural Science (长春师范学院学报),2006,25(5):58-60.

[18]LIANG B, WANG LM, ZHOU L, et al.Induced apoptotic effects of Herba Saginae Japonicae in K562 cell (漆姑草诱导人白血病细胞株K562 凋亡的实验研究)[J]. China Journal of Traditional Chinese Medicine and Pharmacy(中华中医药杂志), 2014, 29(2):571-573.

[19]ZHANG SY(张素英), ZHOU WJ(周万镜). A preliminary screening research on antitumor activity in vitro of extracts in Sagina japonica(Sw.) Ohwi(漆姑草提取物体外抗肿瘤活性的初步筛选研究)[J].Asia-Pacific Traditional Medicine(亚太传统医药),2012,8(8):21-23.

[20]TIAN CG(田成国).Brief report on anticancer drug Sagina japonica (Sw.)Ohwi(抗癌药漆姑草研究简报)[A]. new Chinese medicine (新医学), 1981, 7:360.

[21]HUANG HP(黄厚聘),CHENG CF(程才芬),REN GY(任光友),et al.Research on antitumor effect and preclinical pharmacology of flavonoid glycosides of Sagina japonica(Sw.) Ohwi(漆姑草黄酮苷的抗肿瘤作用与临床前药理研究)[J]. Journl of Guiyang Medical College(贵阳医学院学报),1980,5(1):67-70.

[22]HUANG HP(黄厚聘),CHENG CF(程才芬),LIN WQ(林文琴).Anti-inflammatory effect of flavonoid glycosides of Sagina japonica(Sw.) Ohwi(漆姑草黄酮苷的抗炎作用)[J].Journl of Guiyang Medical College (贵阳医学院学报),1985,10(2):98-99.

[23]HUANG HP(黄厚聘),CHENG CF(程才芬),LI SF (李淑芳),et al.Research onpharmacology of Sagina japonica(Sw.)Ohwi (2)(漆姑草的药理研究2)[J].Journl of Guiyang Medical College (贵州医学院学报),1983A,6(3):1-4.

[24]HUANG HP(黄厚聘),CHENG CF(程才芬),LIN WQ (林文琴),et al. Research on active ingredients of Sagina japonica(Sw.)Ohwi (漆姑草镇咳有效成分研究)[J]. Journl of Guiyang Medical College(贵州医学院学报), 1983B, 8(1):50-51.

[25]EDITORIAL OFFICE OF NATIONAL CHINESE HERBAL MEDICINE COLLECTION(《全国中草药汇编》编写组).Collection of National Chinese Herbal Medicine(全国中草药汇编)(Volume 1)(2ndEdition) [M]. Beijing: People's Medical Publishing House(北京:人民卫生出版社),1996.

[26]INSTITUTE OF BOTANY OF JIANGSU PROVINCE (江苏省植物研究所).Xinhua Materia Medica Outline(新华本草纲要)[M]. Shanghai: Shanghai Science and Technology Press(上海:上海科学技术出版社),1991.

[27]ZHONG XY(钟兴耀),LI H(李恒). Biological characteristics and propagation method of Sagina japonica in Dashanbao(大山包漆姑草的植物学特性及繁殖方法)[J]. Journal of West China Forestry Science (西部林业科学),2005,34(4):97-98.

[28]JIANG LM (蒋利民).Two kinds of effective medicines for treating lacquer dermatitis-Chinese lobelia and Sagina japonica(治疗漆疮的两种良药-半边莲和漆姑草)[J]. Journal of Chinese Lacquer(中国生漆),1983,(4):42-43.

[29]XUE HZ(薛惠彰).The treatment of one case of acute nonlymphocytic leukemia(ANLL) by mainly applying S. japonica(Sw.)Ohwi(漆姑草为主治疗急性非淋巴细胞白血病M2b 型l 例)[J]. Chinese Journal of Integrated Traditional and Western Medicine (中国中西医结合杂志),1994,2:717.

[30]?YANG WP (杨卫平). Common Herbs and Single Proved Recipes for Treatment of Common Illnesses (常用中草药及治疗常见病多发病单验方)[M].Guiyang:Guizhou Nationality Press(贵州科技出版社),2010.

[31]YANG XZ(杨秀珍).A novel lawn grass,Sagina japonica (草坪新秀漆姑草)[J].The Plant Journal(植物杂志),1995,8:22-23.

[32]CAI GX (蔡光先). Materia Medica of Hunan (湖南药物志)[M]. Changsha:Hunan Science and Technology Press(长沙:湖南科技出版社),2014.

[33]WANG Q(王强). Chinese Herbal Medicine Handbook(常用中草药手册)[M]. Fuzhou: Fujian Science and Technology Press(福州:福建科技出版社),2005.

[34]INSTITUTE OF CHINESE MEDICINE OF GUIZHOU PROVINCE (贵州省中医研究所).Guizhou Herbs (贵州草药)[M].Guiyang: Guizhou People's Publishing House (贵州省人民出版社),1970.

[35]YANG H(杨恒),ZHANG H(张辉),SUN J(孙健),et al.Tissue culture of Sagina saginoides from Tibet(西藏无毛漆姑草的组织培养)[J]. Journal of Gansu Agricultural University (甘肃农业大学学报),2011,46(6):66-72.