We prepared protoplasts from Salvia miltiorrhiza Bunge suspension culture cells. Then, the protoplasts' vitality and functions were tested by fluorescein diacetate staining method and Fluo-3/AM flourescent probe. The optimal condition of protoplast isolation was Cellulase R-10 1.5%, Pectinase Y-23 0.3%, Macerozyme R-10 0.5%, 40 r/min 12 h, 600 r/min 5 min, and the protoplasts yield was 1.1x10(6) cells/g FW, the vitality was more than 95% by using fluorescein diacetate staining method. It has been confirmed that calcium fluorescent probe Fluo-3/AM can be successfully loaded into protoplasts.
Aniline Compounds ; chemistry ; Cell Culture Techniques ; Cellulase ; chemistry ; Fluorescent Dyes ; chemistry ; Protoplasts ; chemistry ; Salvia miltiorrhiza ; growth & development ; Xanthenes ; chemistry

Garcinia, a kind of dry resin secreted by Garcinia hanburyi Hook. F. G., is a traditional Chinese medicine with various biological functions such as detoxification, anti-inflammatory, and anthelmintic activities. Recent studies suggest that garcinia has potential anticancer activity. Increasing evidences indicate that the main active monomer gambogic acid isolated from garcinia can inhibit the growth of various cancer cells. Neogambogic acid is an isolated compound with a similar chemical structure as gambogic acid. Preliminary studies show that the neogambogic acid can selectively inhibit the growth of various cancer cells, and has a broader antitumor activity and lower toxicity than gambogic acid. In this review, we summarize the advances made in the investigation of the anti-tumor effect of neogambogic acid in recent years.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; Garcinia ; chemistry ; Humans ; Neoplasms ; drug therapy ; Plant Extracts ; administration & dosage ; chemistry ; Xanthenes ; administration & dosage ; chemistry

OBJECTIVETo develop an HPLC method for determination of components in root of Saposhnikovia divaricata.

METHODThe separation of four components was achieved on a reversed-phase Alltima C18 column with MeOH-H2O gradient elution at a flow rate of 1.0 mL x min(-1).

RESULTSix samples were determined and there is no distinct difference between the contents determined by the new and old methods.

CONCLUSIONThe new method is more simple and convenient than the old method.

Apiaceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Monosaccharides ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Xanthenes ; analysis

Mammalian oocyte maturation and early embryo development processes are Ca(2+)-dependent. In this study, we used confocal microscopy to investigate the distribution pattern of Ca2+ and its dynamic changes in the processes of bovine oocytes maturation, in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development. During the germinal vesicle (GV) and GV breakdown stage, Ca2+ was distributed in the cortical ooplasm and throughout the oocytes from the MI to MII stage. In IVF embryos, Ca2+ was distributed in the cortical ooplasm before the formation of the pronucleus. In 4-8 cell embryos and morulas, Ca2+ was present throughout the blastomere. In PA embryos, Ca2+ was distributed throughout the blastomere at 48 h, similar to in the 4-cell and 8-cell phase and the morula. At 6 h after activation, there was almost no distribution of Ca2+ in the SCNT embryos. However, Ca2+ was distributed in the donor nucleus at 10 h and it was distributed throughout the blastomere in the 2-8 cell embryos. In this study, Ca2+ showed significant fluctuations with regularity of IVF and SCNT groups, but PA did not. Systematic investigation of the Ca2+ location and distribution changes during oocyte maturation and early embryo development processes should facilitate a better understanding of the mechanisms involved in oocyte maturation, reconstructed embryo activation and development, ultimately improving the reconstructed embryo development rate.
Aniline Compounds/chemistry ; Animals ; Calcium/*physiology ; Cattle/*physiology ; Embryonic Development/*physiology ; Female ; Fertilization in Vitro/*veterinary ; Microscopy, Confocal/veterinary ; Oocytes/*physiology ; Parthenogenesis/*physiology ; Xanthenes/chemistry

OBJECTIVETo investigate the xanthones from Tibetan medicine Halenia elliptica and their antioxidant activity.

METHODSColumn chromatography over normal phase silica gel, reversed phase silica gel, Sephadex LH-20, and recrystallization techniques were used to isolate and purify constituents from Halenia elliptica. Infrared spectrometry, mass spectrometry, and nuclear magnetic resonance spectrometry were used to identify the structure of compounds. The antioxidant activity was evaluated by measuring the content of malondialdehyde product in mice liver cell microsomal induced by ferrous-cysteine.

RESULTSEight xanthones (compound I-VIII) were isolated and identified from the ethyl acetate extract of Halenia elliptica, among which 1,7-dihydroxy-2,3,5-trimethoxyxanthone was a novel compound. Compound I, III at 10 microg/ml and 100 microg/ml could inhibit the production of malondialdehyde in mouse liver microsomes in vitro.

CONCLUSIONEight xanthones were isolated and they have certain antioxidant activity.

Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Gentianaceae ; chemistry ; Glycosides ; chemistry ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Xanthenes ; isolation & purification ; pharmacology ; Xanthones ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo investigate the chemical constituents of the brown alga D. divaricata, and to test cytotoxicities of the purified compounds.

METHODCompounds were isolated by normal phase silica gel, Sephadex LH-20 chromatography and reverse phase HPLC techniques. Their structures were elucidated by spectroscopic methods including IR, MS and NMR. Cytotoxicities were tested by MTT method.

RESULTEight compounds were isolated from ethanolic extract of the brown alga D. divaricata and their structures were identified as (-)-torreyol (I), 4beta, 5alpha-dihydroxycubenol (II), 3-farnesyl-p-hydroxybenzioc acid (III), chromazonarol (IV), fucosterol (V), phenyl acetylamine (VI), 4-hydroxybenzoic acid (VII) and n-hexadecanoic acid (VIII).

CONCLUSIONCompound II and IV were obtained from this alga for the first time. The others were isolated from the Dictyotaceae algae for the first time. All compounds were inactive (IC50 > 10 microg x mL(-1)) against human tumor cell lines KB, Bel-7402, PC-3M, Ketr 3 and MCF-7.

Cell Line, Tumor ; drug effects ; Humans ; Parabens ; chemistry ; isolation & purification ; pharmacology ; Phaeophyta ; chemistry ; Stigmasterol ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Terpenes ; chemistry ; isolation & purification ; pharmacology ; Xanthenes ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo develop HPLC methods for the determination of prim-O-glucosylcimifugin and 5-O-methylvisammisoide in Saposhnicovia divaricata and of HPLC fingerprint to compare the wild and culture varieties.

METHODConditions of determination: Shimadzu C18 column (4.6 mm x 150 mm, 5 microm), methanol-water (40:60) as mobile phase with the flow rate of 1 mL x min(-1). The detection wavelength was 254 nm. Conditions of HPLC fingerprint: MG II C18 column (4.6 mm x 250 mm, 5 microm), the mobile phase was acetonitrile-water with the flow rate of 1 mL x min(-1), using linear gradient elution, the column temperature was 30 degrees C.

RESULTThe average recovery of prim-O-glucosylcimifugin was 99.6% (RSD 0.72%, n=6). The average recovery of 5-O-methylvisammisoide was 102.6% (RSD 0.88%, n=6). The contents of prim-o-glucosylcimifugin in wild and culture varieties were (4.96 +/- 2.59) and (3.61 +/- 1.82) mg x g(-1) respectively. The contents of 5-O-methylvisammisoide were (3.91 +/- 2.09) and (4.37 +/- 2.02) mg x g(-1) respectively. The compositions of S. divaricata were effective separated under the conditions of HPLC fingerprint.

CONCLUSIONThe HPLC determination method of prim-O-glucosylcimifugin and 5-O-methylvisammisoide is convenient and accurate. The HPLC fingerprint analysis method could be a basis for quality control and classification evaluate of S. divaricata.

Apiaceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Monosaccharides ; analysis ; isolation & purification ; Quality Control ; Xanthenes ; analysis ; isolation & purification

Fourteen compounds were isolated from the ethanol extraction of Saposhnikovia divaricata (Turcz.) Schischk using column chromatographic methods after enrichment by macroporous adsorptive resins. They were identified as fangfengalpyrimidine (1), clemiscosin A (2), 5-hydroxy-8-methoxypsoralen (3), sec-O-glucosylhamaudol (4), hamaudol (5), nodakenetin (6), prim-O-glucosylcimifugin (7), cimifugin (8), 4'-O-beta-D-glucosyl-5-O-methylvisamminol (9), 5-O-methylvisamminol (10), marmesin (11), adenosine (12), daucosterol (13) and beta-sitosterol (14) by physico-chemical properties and spectral data. Compound 1 is a new compound. Compounds 2 and 3 were isolated from umbelliferae plants and Saposhnikovia divaricata (Turcz.) Schischk for the first time respectively.
Apiaceae ; chemistry ; Chromatography, Thin Layer ; Chromones ; chemistry ; isolation & purification ; Coumarins ; chemistry ; isolation & purification ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Methoxsalen ; analogs & derivatives ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Monosaccharides ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Pyrimidines ; chemistry ; isolation & purification ; Resins, Synthetic ; Xanthenes ; chemistry ; isolation & purification

OBJECTIVETo study the chemical constituents from the stems and leaves of Calophyllum inophyllum.

METHODThe compounds were isolated by column chromatography on silica gel, Sephadex LH-20 and preparative TLC. Their structures were elucidated by chemical methods and NMR, MS spectroscopic data.

RESULTNine compounds were identified as 2-hydroxyxanthone (1), 4-hydroxyxanthone (2), 1, 5-dihydroxyxanthone (3), 1, 7-dihydroxyxanthone (4), 1, 3, 5-trihydroxy-2-methoxyxanthone (5), 6-deoxyjacareubin (6), amentoflavone (7), kaempferol-3-O-alpha-L-rhamnoside (8) and quercetin-3-O-alpha-L-rhamnoside (9).

CONCLUSIONCompounds 8 and 9 were isolated from the genus Calophyllum and compounds 1, 2, 4-6 were isolated from this plant for the first time.

Calophyllum ; chemistry ; Chromatography ; methods ; Flavonoids ; analysis ; chemistry ; isolation & purification ; Glycosides ; analysis ; chemistry ; isolation & purification ; Kaempferols ; analysis ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; methods ; Mass Spectrometry ; methods ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Pyrans ; analysis ; chemistry ; isolation & purification ; Quercetin ; analogs & derivatives ; analysis ; chemistry ; isolation & purification ; Xanthenes ; analysis ; chemistry ; isolation & purification

The extraction of active ingredient group, i.e., prim-o-glucosylcimifugin, astragaloside, and 5-o-methylvisammiosode from Yupingfeng powder with one-pot method was studied in this work. A HPLC method was used to determine the content of each active constituent mentioned above in the extract. The influences of extraction temperature, time, volume percent of ethanol in water and its amount added on the content and yield of active ingredient group were investigated by orthogonal test. The experimental results showed that the optimized extraction conditions were as follows: 1g of Yupingfeng powder was one-pot extracted for 4 hours at 80 degrees C with 90% ethanol as solvent, and the yield and content of active ingredient group were 0.16%, 0.53% respectively. The active ingredient group in Yupingfeng powder could be effectively one-pot extracted under the conditions above.
Apiaceae ; chemistry ; Astragalus membranaceus ; chemistry ; Atractylodes ; chemistry ; Chemical Fractionation ; methods ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Monosaccharides ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; Powders ; Saponins ; analysis ; isolation & purification ; Temperature ; Triterpenes ; analysis ; isolation & purification ; Water ; chemistry ; Xanthenes ; analysis ; isolation & purification