[Normal light and fluorescence microscopy for authentication of Delphinii Brunoniani Herba of Tibet].

Dried herb of Delphinium brunonianum Royle (Ranunculaceae) has long been used under the herbal name "Xiaguobei" (Delphinii Brunoniani Herba) in traditional Tibetan medicine and prescribed for the treatment of influenza, itchy skin rash and snake bites. In order to find a useful and convenient method for the identification of microscopic features, the technique of fluorescence microscopy was applied to authenticate "Xiaguobei" of Tibet. The transverse sections of stem and leaf, as well as the powder of "Xiaguobei" were observed to seek for typical microscopic features by normal light and fluorescence microscopy. A style-like, single-cell glandular hair containing yellow secretions on the leaf, young stem and sepal of "Xiaguobei" was found. Under the fluorescence microscope, the xylem and pericycle fiber group emitted significant fluorescence. This work indicated that fluorescence microscopy could be an useful additional method for the authentication work. Without the traditional dyeing methods, the main microscopic features could be easily found by fluorescence microscopy. The results provided reliable references for the authentication of "Xiaguobei".

Five new compounds from Dendrobium crystallinum.

Five new compounds, dencryol A (1), dencryol B (2), crystalltone (3), crystallinin (4), and 3-hydroxy-2-methoxy-5,6-dimethylbenzoic acid (5), together with six known compounds, dendronobilin B (6), syringic acid (7), apigenin (8), isoviolanthin (9), 6'''-glucosyl-vitexin (10), and palmarumycin JC2 (11), have been isolated from the stems of Dendrobium crystallinum, of which compounds 9-11 were isolated from the genus Dendrobium for the first time, and all the other compounds were first obtained from this plant. Their structures were established on the basis of spectroscopic analysis and literature data.

[Phenolic components from herbs of Dendrobium aphyllum].

OBJECTIVE: To study the phenolic constituents of Dendrobium aphyllum. METHOD: Various chromatographic techniques were used to isolate and purify the constituents, their physico-chemical properties and spectral data were employed to elucidate their structures. RESULT: Nine bibenzyls and two benzylethanyl compounds were isolated and identified as: moscatilin (1), gigantol (2), batatasin (3), tristin (4), 3, 5, 4'-trihydroxylbibenzyl (5), 3, 5-dimethoxyl-4, 4'-dihydroxylbibenzyl (6), moscatin (7), 2, 4, 7-trihydroxyl-9, 10-dihydrophenanthrene (8), hircinol (9), 2-(4-hydroxyphenyl) ethyl-beta-D-glucopyranoside, salidroside (10) and p-hydroxylbenzylacetic acid (11). CONCLUSION: All compounds were obtained firstly from the plant, and the compounds 10 and 11 were isolated in this genus for the first time.

[Studies on chemical constituents of Dendrobium crystallinum].

OBJECTIVE: To study the chemical constituents of Dendobium crystallinum. METHOD: Compounds were isolated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were identified by physicochemical properties and spectral analyses. RESULT: Nine compounds were obtained and identified as: 4, 4'-dihydroxy-3, 5-dimethoxybi-benzyl (1), gigantol (2), naringenin (3) , p-hydroxybenzoic acid (4), n-tetracosyl trans-p-cou-marate (5), n-octacosy trans-p-coumarate (6), n-hexacosyl trans-ferulate (7), stigmasterol (8), daucosterol (9). CONCLUSION: All these compounds were obtained from this plant for the first time, compounds 1 and 4 were isolated firstly from the genus.

[Studies on chemical constituents in stem of Dendrobium chrysotoxum].

OBJECTIVE: To investigate the chemical constituents of Dendrobium chrysotoxum. METHOD: The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by spectral evidences. RESULT: Ten compounds were obtained and identified as (+)-syringare sinol (1), 5alpha, 8alpha-epidioxy-24( R)-methycholesta-6, 22-dien-3beta-ol (2), trans-3-(4-hydroxy-3-methoxyphenyl)-acrylic acid octacosyl ester (3), defusin (4), 3, 4-dihydroxy benzoic acid (5), 3, 4-dimethoxy-benzoic acid (6), vanillic acid (7), 3, 4-dimethoxy-benzoic acid methyl ester (8), 3, 5-dibromo-2-aminobenzaldehyde (9), heptadecanoic acid 2, 3-dihydroxy-propyl ester (10). CONCLUSION: Compounds 1, 2 and 6-10 were isolated from this plant for the first time.

Bi-bicyclic and bi-tricyclic compounds from Dendrobium thyrsiflorum.

One bi-bicyclic and two bi-tricyclic derivatives of coumarin-benzofuran, phenanthrene-phenanthrene and phenanthrene-phenanthraquinone, along with seven known compounds, were isolated from stems of Dendrobium thyrsiflorum Rchb.f. (Orchidaceae). On the basis of chemical, NMR (1H, 13C, HMQC, HMBC and NOESY) and mass spectrometry data, their structures were elucidated as denthyrsin [3-(5',6'-dimethoxybenzofuran-2'-yl)-6,7-dimethoxy-2H-chromen-2-one; 1], denthyrsinol (4,5'-dimethoxy-[1,1']biphenanthrenyl-2,5,4',7'-tetraol; 2), and denthyrsinone (7,4',7'-trihydroxy-2,2',8'-trimethoxy-[5,1']biphenanthrenyl-1,4-dione; 3). Compounds 1-3 and denthyrsinin (1,5,7-trimethoxyphenanthrene-2,6-diol; 4) showed significant cytotoxic activities against Hela (13.5, 9.3, 9.9 and 2.7 microM, respectively), K-562 (0.45, 1.6, 6.0 and 2.3 microM, respectively) and MCF-7 (18.1, not tested, 3.5 and 4.8 microM, respectively) cell lines.

[Location and relative quantity of coumarins in the stem of Dendrobium thyrsiflorum].

AIM: To determine the location and relative quantity of coumarins in the stem of Dendrobium thyrsiflorum Rchb. f. , and to provide a scientific basis for evaluating and utilizing the famous medicinal plant. METHODS: The stems of one, two and three years old, separately, were collected in February. Location and relative quantity of coumarins in the top, middle and basal parts of each stem sample were determined by using laser scanning confocal microscopy (LSCM). ANOVA and Tukey's test were employed in the statistical analysis. RESULTS: The coumarins located mainly in vascular bundles, especially on the walls of the outer fiber cells. There was significant difference or great significant difference exist in every part of the stem from different ages. The values of significant difference were as follows: basal part P = 0.004 (< 0.01); middle part P = 0.009 (< 0.01); top part P = 0.036( < 0.05). CONCLUSION: Two years old stem could be the best choice when collecting Dendrobium thyrsiflorum in Feburary.

[Molecular identification of medicinal plants: Dendrobium chrysanthum, Dendrobium fimbriatum and their morphologically allied species by PCR-RFLP analyses].

AIM: To establish a simple method for molecular identification of original plants of D. chrysanthum and D. fimbriatum using molecular marker rDNA ITS region. METHODS: Restriction patterns of ITS fragments were obtained using PCR-RFLP method. The PCR products of D. chrysanthum and its morphologically allied species were digested at 37 degrees C by Cla I and Apa LI, those of D. fimbriatum and its morphologically allied species were digested by Sph I. RESULTS: D. chrysanthum, D. fimbriatum and their morphologically allied species could be identified by predicted restriction profiles of PCR-RFLP. The botanical origin of twenty-five fresh samples of "Shihu" collected in markets was identified by this method. CONCLUSION: The results showed that PCR-RFLP analysis of the rDNA ITS region is a feasible, simple and inexpensive method for determining the botanical origin of the traditional Chinese medicine "Shihu".

[Analysis on the geologic background and physicochemical properties of soil for the cultivation of Panax notoginseng in Yunnan province].

OBJECTIVE: To study the relationship between the geo-authentic and geologic background system (GBS) and physicochemical properties of soil for the cultivation of Panax notoginseng. METHOD: The physicochemical properties of soil were analyzed, the geologic background system between producing areas of Yunnan and Guanxi of P. notogiseng was compared. RESULT: The distribution of P. notoginseng was restricted by GBS, The yellow-red soil of fragmentary rock mixed with carbonate seems to be most suitable for the growth. CONCLUSION: The most suitable soil for growing P. notoginseng is the neutral or weakly acidic soil with loam clay, particularly that with low base saturation percentage.

[Studies on the chemical constituents of Dendrobium fimbriatum].

AIM: To investigate the chemical constituents of Dendrobium fimbriatum Hook. METHODS: Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by IR, MS, 1HNMR, 13CNMR and 2D-NMR. RESULTS: Eight compounds were obtained and identified by spectral analysis as fimbriatone, confusarin, crepidatin, physcion, rhein, ayapin, scopolin methyl ether and n-octacostyl ferulate. CONCLUSION: Fimbriatone is a new compound, physcion and rhein are firstly isolated from Genus Dendrobium. The others are found from this species for the first time. Fimbriatone showed potential inhibitory effects on BGC cell line.

[RbcL sequence analysis of Belamcanda chinensis and related medicinal plants of Iris].

AIM: To identify "Shegan" [Belamcanda chinensis (L.) DC.] and relative medicinal plants of Iris including Iris tectorum Maxim., I. dichotoma Pall., I. germanica L. and I. japonica Thunb. by ribulose 1,5-bisphosphate carboxylase Large Gene (rbcL) sequence analysis. METHODS: General DNA was isolated from the fresh leaves of Belamcanda chinensis and 4 Iris spp. by CTAB. A pair of primers was designed to amplify the rbcL gene and PCR Preps DNA kit was used to purify the PCR products. The rbcL sequences were determined by ABI (Applied Biosystems Inco.) Prism 310 Genetic Analyzer. RESULTS: A fragment of about 750 bp of rbcL gene from Belamcanda chinensis and 4 Iris spp. were amplified and sequenced. The rbcL sequences of Iris tectorum, I. dichotoma Pall. and I. japonica were reported for the first time. The rbcL sequences of 5 species of Iridaceae were aligned and analyzed using Clustal (Version 8.0) and MEGA (Version 2.0.) programs. The nucleotide number of difference is from 1.000 to 20.000. The tranversions is from 0.000 to 9.000 and the transitions is from 0.000 to 14.000. Phylogenetic tree based on rbcL partial sequence data indicated that the eleven samples of 5 species clustered separately. CONCLUSION: The sequence variation of rbcL can be used to identify Belamcanda chinensis and 4 species of relative medicinal plants of Iris. The molecular phylogenetic tree accords with the classical taxonomy.

[Pharmacognostical identification of "guijiu ([symbol: see text])" in Shosoin of Japan].

OBJECTIVE: To identify the botanical origin of the Guijiu in Shosoin of Japan from Tang Dynasty, and trace its medicinal history. METHOD: Anatomical characteristics of the underground parts of Guijiu in Shosoin were compared with those of Hosta plantaginea and H. ventricosa, and research on the medicinal history of Guijiu was made based on its original identification results and describes in herbals. RESULT AND CONCLUSION: Guijiu in Shosoin of Japan was derived from the underground parts of H. plantaginea and is one of Guijiu used in Tang Dynasty and earlier on.

[Studies on chemical constituents of Pholidota yunnanensis].

OBJECTIVE: To investigate the chemical constituents from Pholidota yunnanensis. METHOD: Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical and spectral analyses. RESULT: Seven compounds were obtained and they were identified by spectroscopic analysis as n-nonacosane, cyclopholidone, n-dotriacontanoic acid, n-octacostyl ferulate, cyclopholidonol, cycloneolitsol and beta-sitosterol, respectively. CONCLUSION: n-octacostyl ferulate and cycloneolitsol were isolated from genus Pholidota for the first time.

[Studies on chemical constituents of two plants from Costus].

OBJECTIVE: To study the chemical constituents of Costus speciosus and C. tonkinensis (Zingiberaceae) distributed in Yunnan province. METHOD: Chromatography and spectral analyses were used to isolate the constituents and elucidate their structure. RESULT: Six compounds were isolated from the rhizome of C. speciosus and elucidated as diosgenin(1), prosapogenin B of dioscin(2), diosgenone(3), cycloartanol(4), 25-en-cycloartenol(5) and octacosanoic acid(6). Four compounds were isolated from the rhizome of Costus tonkinensis and elucidated as tetracosanoic acid(7), succinic acid(8), beta-sitosterol(9) and daucosterin(10). CONCLUSION: Compounds of 3-6 were obtained from C. speciosus for the first time and compounds of 7-10 were obtained from C. tonkinensis for the first time too.

[Studies on chemical constituents of Alpinia jianganfeng].

OBJECTIVE: To study the chemical constituents of Alpinia jianganfeng (Zingiberaceae) distributed in Sichuan province. METHOD: Chromatography and spectral analyses were used to isolate the constituents and elucidate their structure. RESULT: Four compounds were isolated from the rhizome of A. jianganfeng and elucidated as kaempferol-3-O-glucuronide(1), docosanoic acid(2), 3-hydroxy-stigmast-5-en-7-one(3) and beta-sitosterol(4). CONCLUSION: All these compounds were obtained from this plant for the first time.

[Determination of linderane in root tuber of Lindera aggregata by HPLC].

OBJECTIVE: To provide scientific basis for quality control of Lindera aggregata. METHOD: HPLC analytical method was established using a Lichrospher C18 column and acetonitrile-water (56:44) as the mobile phase, detected at 235 nm. RESULT: The linear range of linderane is between 0.0642 - 0.5774 microg, the average recovery was 98.4%, RSD1.7% (n = 9). CONCLUSION: Contents of linderane in commercially available and collected samples were from 0.028% to 0.123% and from 0.056% to 0.222% respectively.

[Study on sequence difference and SNP pheomenon of rDNA ITS region in F type and H type population of Dendrobium officinale].

OBJECTIVE: To study rDNA ITS sequence differences between F type and that of H type of Dendrobium officinale in main habitat of China. METHOD: The population differences of the rDNA ITS region (including ITS1, ITS2, 5.8S) sequences of D. officinale were studied by the method of DNA sequences analysis. RESULT: There were two different sites between the rDNA ITS sequence of F type and that of H type. One was in ITS1 region, and the other was in 5.8S region. It was proved that there was some relativity between the character of rDNA ITS region and the life type of the populations. The phenomenon of single nucleotide polymorphism (SNP) existed in 5.8S region of rDNA ITS region between F type and H type. The sequences of rDNA ITS region of D. officinale were reported for the first time, and the sequences of ITS region ranged 634 bp (ITS1 231 bp, ITS2 240 bp, 5.8S 163 bp). CONCLUSION: The analysis of rDNA ITS of D. officinale deeply reveal the population differences of D. officinale of F type and H type.

[Analysis of 1H-NMR fingerprint in stem of Dendrobium loddigesii].

OBJECTIVE: To analyse the 1H-NMR finger-print of the stem of Dendrobium loddigesii. METHOD: Silica gel column chromatography was used to separate the chemical constituents of SCE A of the stem of D. loddigesii. The characteristic signals of the H-NMR finger-print were analysed after determining the structures of the compounds isolated from SCE A. RESULT: 1H-NMR finger-prints of the samples of D. loddigesii collected from different regions showed highly characteristic features and reproducibility. Four compounds predominant in SCE A were isolated and their structures were determined by spectral analysis as 1, 2, 3 and 4, respectively. CONCLUSION: Compound 3 and 4 were isolated from D. loddigesii for the first time. The 1H-NMR finger-print of CGE A of the stem of D. loddigesii showed mainly the characteristic signals of the above four compounds and might be utilized for the original authentication of this plant.

Two new dammarane triterpene glycosides from the rhizomes of Panax notoginseng.

Two new dammarane triterpene glycosides named notoginsenosides Rw 1 (1) and Rw 2 (2) were isolated from the rhizomes of Panax notoginseng, together with 20 known compounds including protopanaxadiol (3), protopanaxatriol (4), ginsenosides Rb1 (5), Rd (6), Re (7), Rg1 (8), Rg2 (9), 20-(S)-Rg3 (10), 20-(R)-Rg(3) (11), Rh1 (12), Rh4 (13), Rf (14), 20-O-glucopyranosyl Rf (15), notoginsenosides R1 (16), R2 (17), T5 (18), S (19), T (20), and Fa (21), and koryoginsenoside R1 (22). Based on FABMS, HRFABMS, IR, (1)H, 13C, and 2D-NMR (HSQC, HMBC, and COSY) spectral data, the structures of the new compounds were elucidated as 6-O-beta-d-xylopyranosyl-20-O-beta-d-xylopyranosyl-(1 --> 6)-beta-d-glucopyranosyldammar-24-ene-3beta,6alpha,12beta,20(S)tetraol (1) and 6-O-beta-d-xylopyranosyl-(1 --> 2)-beta-d-glucopyranosyldammar-22-ene-(trans)-3beta,6alpha,12beta,20(S), 25-pentaol (2). Compounds 3, 4, 13, and 22 were isolated from P. notoginseng for the first time.

A new phenanthrene with a spirolactone from Dendrobium chrysanthum and its anti-inflammatory activities.

Investigation of phenolic patterns from the stems of Dendrobium chrysanthum by HPLC-PDA-MS has led to the isolation of a new phenanthrene derivative with a spirolactone ring, dendrochrysanene (1), that proved to suppress the mRNA level of TNF-alpha, IL8, IL10, and iNOS in murine peritoneal macrophages. The structure of 1 was characterized on the basis of various NMR (1H, 13C, 1H-1H COSY, HMQC, and HMBC), mass spectrometry, and X-ray crystal diffraction data.