Flavonoids from Epimedium wanshanense.

A novel flavonol glycoside named wanepimedoside A was isolated from the whole plant of Epimedium wanshanense, along with fifteen known flavonoids, anhydroicaritin, desmethylanhydroicaritin, icarisid I and II, quercetin, ikarisoside A and B, sagittatoside B, 2"-O-rhamnosylicarisid II, icariin, 2"-O-rhamnosylikarisoside A, epimedin B, epimedin C, and diphylloside A and B.

Xanthones from Hypericum japonicum and H. henryi.

From the aerial part of Hypericum japonicum, one new xanthone glycoside, 1,5-dihydroxyxanthone-6-O-beta-D-glucoside, one novel dimer xanthone, bijaponicaxanthone, and the first natural prenylated xanthone, 1,3,5,6-tetrahydroxy-4-prenylxanthone, were isolated together with the four known xanthones, 1,5,6-trihydroxyxanthone, isojacereubin, 6-deoxyisojacareubin and 4',5'-dihydro-1,5,6-trihydroxy-4',4',5'-trimethylfurano (2',3':4,5) xanthone. five previously known xanthones, kielcorin, cadensin, 1,7,-dihydroxyxanthone, 1,5-dihydroxy-4-methoxyxanthone and 1,2,5-trihydroxyxanthone were also found in the dichoromethane extract of the stems and leaves of H. henryi. Their structures were elucidated by spectroscopic and chemical methods. Some of the compounds from H. japonicum were found to exert an interesting coagulant activity in an in vitro test. The chemotaxonomic value of xanthones is discussed briefly.

Flavonol glycosides from Epimedium koreanum.

A novel flavonol glycoside named caohuoside-B was isolated from the aerial parts of Epimedium koreanum, along with a known flavonol glycoside, epimedokoreanoside-I. Their structures were established by spectroscopic methods. The new compound was elucidated as anhydroicaritin 3-O-beta-D-(4,6-O- diacetyl)glucopyranosyl-(1-->3)-alpha-L-(4"-O-acetylrhamnopyranoside+ ++)-7-O-beta- D-glucopyranoside.

Anhydroicaritin 3-O-rhamnosyl(1-->2)rhamnoside from epimedium koreanum and a reappraisal of other rhamnosyl(1-->2, 1-->3 and 1-->4)rhamnoside structures.

An anhydroicaritin 3-O-rhamnosylrhamnoside from Epimedium koreanum is defined as the 3-O-alpha-L-[alpha-L-rhamnopyranosyl(1-->2)rhamnopyranoside] on the basis of 2D NMR evidence. Complete assignments of the 1H and 13C NMR spectra of this compound are presented for the first time. The NMR distinction of 1-->2, 1-->3 and 1-->4 linked rhamnopyranosylrhamnopyranosides are discussed and indicate that baohuosides III and V from E. davidii and baohuoside VI from E. davidii and E. pubescens, respectively, are (1-->2) linked.

An unusual cycloartane triterpenoid from Cimicifuga foetida.

A new cycloartane triterpenoid glycoside has been isolated from the rhizomes of Cimicifuge foetida L. The spectroscopic characteristics of the new compound are different from previously described cycloartane triterpenoids because of the loss of the 24-isopropyl group as well as the presence of a 11 beta-OH group. Based on spectroscopic evidence, including a series of 2D-NMR analyses, the structure of the new triterpene is assigned as 24-des-isopropyl-7-ene-23-one-9,19; 16,24-dicycloart-3 beta,11 beta,16 alpha,24 alpha-tetraol 3-O-beta-D-xylopryanoside, named here as neocimiside. The structure of the aglycone of neocimiside was confirmed by X-ray analysis.

Ethnopharmacologic investigation on tropane-containing drugs in Chinese solanaceous plants.

The role of tropane-containing drugs derived from Solanaceae has long been recognized. This report deals with Chinese solanaceous plants, including historic, botanical, phytochemical, pharmacological and clinical aspects. Our investigation revealed that this kind of drug is constantly being confirmed to be of true therapeutic value; however, further studies are in order.

Recent advances on ginseng research in China.

Ginseng, Panax ginseng C.A. Meyer, is a well-known Chinese traditional medicine. There have been more than 300 original papers in Chinese and in English during the last 10 years in China. This review paper summarizes some achievements from some of these published papers. Twenty-eight ginsenosides and some minor constituents were extracted and isolated from the root, root-stock, stems, leaves, flowers and flower-buds of ginseng. The chemical analysis demonstrated that the content of ginsenosides is related to the source, part and growth years of ginseng. The drug has a wide range of pharmacological and therapeutical actions, it acts on the central nervous system, cardiovascular system and endocrine secretion, promotes immune function and metabolism, possesses biomodulation action, anti-stress and anti-ageing activities, and so on. Many preparations of ginseng have been officially approved for clinical application in China. Clinical evaluation has shown that these preparations play a special role in medicinal use.

Constituents of Chinese Piper species and their inhibitory activity on prostaglandin and leukotriene biosynthesis in vitro.

The n-hexane extracts of 19 Piper species, predominantly from China, were screened for their 5-lipoxygenase (5-LOX) and cyclooxygenase-1 (COX-1) inhibitory potential. Many of them showed considerable inhibitory activity against at least one of these two key enzymes of the arachidonic acid metabolism, especially against COX-1. The best results in inhibiting the formation of leukotrienes were obtained with the extract of Piper kadsura. In the terms of prostaglandin synthesis inhibition, the extract of Piper boehmeriifolium var. tonkinense was found to have the strongest activity. Furthermore, an analytical investigation by means of TLC, HPLC-DAD and GC-MS resulted in the identification of 20 constituents. Most of them were amides with an interesting variety of amine moieties. Among them were pellitorine, and four higher homologues, piperlonguminine, dihydropiperlonguminine, futoamide, chingchengenamide, the retrofractamides A, B and D, guineensine, brachystamide B, piperanine, piperine, piperdardine, sarmentine, pipataline and benzylbenzoate. In 96 cases, these constituents were new for the particular plant.

Immunological aspects of Chinese medicinal plants as antiageing drugs.

The development of a predominantly geriatric community worldwide has become an inevitable fact. Antiageing agents could be, in a certain sense, attentive to the well-being of the aged. There are quite a lot of medicinal plants and prescriptions recorded in Chinese medical literatures aimed at the well-being of the aged as well as the prevention of diseases and prolongation of life-span. By means of modern scientific research, a strategy towards antiageing drugs is presented in this paper. One of the effective routes is to select the candidates based on their ethnopharmacological usages, followed by immunological investigation in connection with other antiageing experimentation. A list of Chinese medicinal plants used as or related to the antiageing agents are presented. Specifically, five Chinese traditional drugs, Herba Epimedii, Fructus Lycii, Radix Polygoni multiflori, Radix Cynanchi auriculati and Ganoderma along with a composite prescription 'American Ginseng Royal Jelly' are selected as representatives. The prospect of research and development of antiageing drugs based on natural origin is also discussed.

Effect of berbamine on T-cell mediated immunity and the prevention of rejection on skin transplants in mice.

Berbamine, an ingredient of Berberis, which itself is widely utilized in Chinese folk-medicine has been used as a source of leukogenics, anti-arrhythmics and anti-hypertensives. In recent years, the immunosuppressive effects of berbamine has been demonstrated. In order to further investigate the value of berbamine as an immunosuppressive agent, the delayed type hypersensitivity reaction (DTH) response with sheep red blood cells (SRBC), the mixed lymphocyte reaction (MLR) and a skin model of allograft rejection on mice were studied. Berbamine showed suppressive effects on DTH and MLR and significantly prolonged allograft survival compared with untreated transplanted mice. The results indicate that berbamine may be a potential agent in clinical transplantation.

[HPTLC scanning determination of 6 flavonoids in 166 Rhododendron species].

This paper reports a quantitative determination in the contents of flavonoids in Rhododendron leaves by HPTLC scanning method. The methanol extract of each sample was spotted on a HPTLC silica gel plate (E. Merck F254) alongside with standard substances. Using the upper layer of petroleum ether (60-90 degrees C)-ether-formic acid (62:31:7) as developing solvent I, farrerol, kaempferol and quercetin were well separated. The spots were determined by a single wavelength (366 nm) TLC-scanner, Camag model 76510. The same plate was then further developed by developing solvent II:7 ml of the lower layer of chloroform-methanol-water (7:3:1) plus 0.5 ml of formic acid to separate polystachoside, quercitrin and hyperoside. The densitometric determination of each spot was also carried out with the same scanner. The contents of 6 flavonoids in the leaves of 166 Rhododendron species were thus determined.

[Research on DNA molecular marker of Magnolia officinalis Rehd. et Wils.--RAPD study on certified species].

AIM: To discuss the intraspecific relationship in Magnolia officinalis and the genuineness of Cortex Magnoliae officinalis, and to find some DNA characters of certified "Houpo". METHODS: Thirty-three samples from eleven locations, which can represent most of the distribution of M. officinalis, were selected. The total DNA was extracted. Severty-four random primers were tried to get good amplification. RESULTS: One hundred and sixteen bands amplified from seventeen primers, were clustered by NTSYS-pc software. Three branches were obtained. Some distinctive primers and bands, which represent certified species or fine breed, were obtained also. CONCLUSION: 1) M. officinalis should be divided into three geographic clans instead of two subspecies or varieties, they are, a) typical officinalis, b) typical biloba and c) Middle type. This conclusion agrees with the leaf form and other characters. 2) The genetic difference between "Chuanpo" and "Wenpo" is evident and the difference is in correspondence with the quantities of their chemical constituents. So, the genetic difference is the main reason of the genuineness of Cortex Magnoliae officinalis. 3) These results may be used to establish DNA database for identification of Cortex Magnoliae officinalis.

[Studies on chemical constituents of the essential oil of Cyclorhiza waltonii].

Essential oil from the rhizome of Cyclorhiza waltonii was obtained by steam distillation. The chemical components of the oil were examined by means of capillary gas chromatography and gas chromatography-mass selective detector with HP-5 and Carbowax 20M stationary phases. Fifteen constituents of the oil were identified. The content of each component identified was determined by area normalization method. Of these, the content of ten compounds was higher than 2%. They are: alpha-pinene, sabinene, alpha-therpinene, p-cymene, beta-phellandrene, gamma-terpinene, terpinolene, terpineo-4-ol, cis-caryophyllene, duryl aldehyde and isoduryl aldehyde. The oil was found to compose of almost all the types of volatile composition, i.e., not only monoterpenes and sesquiterpenes but also alcohols and aldehydes. Duryl aldehyde and isoduryl aldehyde are not common in volatile oils.

[Determation of strychnine and brucine in Strychnos by HPLC].

This paper describes the determination of strychnine and brucine in the seeds, root, stem and leaves of Strychnos species by HPLC. The analytical column used was ZY110 YNG-C18. The mobile phase was KH2PO4(0.01 mol.L-1)--MeOH(73:27), pH2.5, regulated by 10% H3PO4. Flow rate was 1.0 ml.min-1. The detection wavelength was 264 nm. The linear ranges of strychmine and brucine were 0.18-7.26 micrograms and 0.11-4.32 micrograms, respectively. The recoveries of strychnine and brucine were 98.27% and 98.04%, respectively. The analytical results showed that the contents of strychnine and brucine in samples showed great difference between different species. The contents of strychnine in the seeds of Strychnos wallichiana and S. ignatii were 5.6% and 3.9%, respectively. These results show that the two Strychnos species may be developed as the resources of strychnine.

[Studies on polysaccharides in different development stages of Armillaria mellea (Vahl.: Fr.) Quel].

OBJECTIVE: To study the polysaccharides in different development stages of Armillaria mellea. METHOD: Polysaccharides in rhizomorph, fruit-body, mycelia and its fermenting liquor of A. mellea were extracted, isolated and purified. The properties contents, molar ratio and molecular weight of the polysaccharides were determined by IR spectra, HPLC, GPC and gel chromatography. RESULT: The polysaccharides in mycelia and its fermenting liquor contained only glucose, the sugar contents were 9.00% in mycelia and 0.87 g.(100 ml)-1 in fermenting liquor respectively. The polysaccharides in both rhizomorph and fruit-body consisted of glucose and xylose, and the molar ratio was 1:14 in rhizomorph and 1:10 in fruit-body respectively; the polysaccharide contents were 1.12% in rhizomorph and 2.27% in fruit-body. The molecular weight of these polysaccharides was about 10,000-70,000. CONCLUSION: The experimental results supply important scientific data for developing A. mellea as a medicine.

[Comparison of ITS sequences between wild ginseng DNA and garden ginseng DNA].

OBJECTIVE: To compare the genetic differences between wild ginseng and garden ginseng (Panax ginseng). METHOD: The sequences of ITS1 and ITS2 of wild ginsengs were determined on LKB DNA sequencing station through Si-liver Sequence DNA Sequencing System. The sequencies were aligned with DNA SIS software. RESULTS AND CONCLUSION: The ITS1 and ITS2 of Panax were 220-221 and 222-224 bases in length respectively. In Panax ginsehg, the seqences of ITS1 were very stable, but ITS2 were changeable. The ITS2 sequences of No. 87 and No. 110 of the wild ginseng collected from Fusong Heilongjiang (China) were exactly the same as those of No. U41680(Jun Wen) and No. U41682(Jun Wen) of garden ginseng collected from Heilongjiang Province (China) and Korea respectively, but different from those of No. U41681(Jun Wen) from Hubei Province (China) in three bases (447, 449, 450) The result implies that the cultivated ginsengs may have been introduced from two different populations of the wild ginseng.

[A study on AFLP fingerprinting of land races of Panax ginseng L].

OBJECTIVE: To obtain more information on DNA fingerprintings of five land races of Chinese ginseng, namely, Damaya (DMY), Changbo (CB), Yuanbangyuanlu (YBYL) and Huangguo (HG). METHODS: The five land races were detected by amplified restriction fragment polymorphism (AFLP) markers with 11 combined primers (M2, M3, M16, M20, M53, M56, M57, M68, M69, M72, M84 in Mse I). RESULT AND CONCLUSION: Only 4.6% polymorphic sites was found. It was further verified that only a little diversity existed among the land races. The polymorphic sites of CB were much more than those of the others, which suggests that there are more heterozygotes in CB populations, and it is closer to wild ginseng than the others.

[Studies on the chemical constituents from Aquilegia ecalcarata Maxim].

OBJECTIVE: To study the chemical constituents from Aquilegia ecalcarata. METHOD: Compounds were isolated with silica gel and polyamide chromatography and their structures were determined by spectral analysis and chemical evidence. RESULT: Five compounds were obtained and identified as beta-sitosterol, isoorientin-7-O-glucoside, isovitexin-4'-O-glucoside, isovitexin-2'-O-rhamnoside, luteolin-7-O-glucoside. CONCLUSION: All the compounds were isolated from this plant for the first time.

Sequence characteristics and divergent evolution of the chloroplast psbA-trnH noncoding region in gymnosperms.

The psbA-trnH intergenic region is among the most variable regions in the gymnosperm chloroplast genome. It is proposed as suitable for DNA barcoding studies and is useful in phylogenetics at the species level. This region consists of two parts differing in their evolutionary characteristics: 1) the psbA 3’UTR (untranslated region) and 2) the psbA-trnH intergenic spacer. We compared the sequence and RNA secondary structure of the psbA 3’ UTR across gymnosperms and found consensus motifs corresponding to the stem portions of the RNA stem-loop structures and a consensus TGGATTGTTATGT box. The psbA-trnH spacer is highly variable in length and composition. Tandem repeats that form stem-loop structures were detected in both the psbA 3’ UTR and the psbA-trnH spacer. The presence of promoters and stem-loop structures in the psbA-trnH spacer and high sequence variation in this region suggest that psbA and trnH in some gymnosperms are independently transcribed. A comparison of chloroplast UTRs across gymnosperms offer clues to the identity of putative regulatory elements and information on selective constraints imposed on the chloroplast non-coding regions. The present study should inspire researchers to explore the full potential of the psbA-trnH non-coding sequence and to further stimulate its application in a broader spectrum of studies, not limited to phylogenetics and DNA barcoding.