Objective To investigate the mechanisms of oridonin inducing apoptosis on acute leukeamia NB4 cells and its mechanism. Methods NB4 cells in culture medium in vitro were given with different concentrations (8, 16, 24, and 32 μmol/L) of oridonin. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 expression was detected by Western blotting, and caspase-3 activity was assayed with colorimetric assay kit before and after apoptosis occurred. Results Oridonin (over 16 μmol/L) could inhibit the growth of NB4 cells and cause apoptosis significantly, the suppression was both in a timeand dose-dependent manner. Marked changes of apoptosis including condensation of chromatin and nuclear fragmentation were observed very clearly by Hoechst 33258 fluorescence staining and a characteristic "ladder" of DNA fragments was elicited by agarose gel electrophoresis; Western blot analysis revealed that caspase-3 was activated by the loss of caspase-3 proenzyme (32 kDa) and the appearance of its 20 kDa subunit, and that along with the apoptotic process caspase-3 activity was increased concurrently. Conclusion Oridonin can induce apoptosis in NB4 cells via activation of caspase-3. These results will provide laboratory evidence for the clinical treatment of acute leukemia with oridonin.

Objective To fully understand the medicinal plants of Fritillaria L. , the physicochemical properties of starch in two species of Fritillaria L. , F. thunbergii and F. ussurensis. were investigated by means of various analytical methods. Methods The properties of starch in the two different species of Fritillaria L. were compared by X-ray diffraction, scanning electron microscope (SEM) and themogravimetric analysis (TGA). Results The crystal type of starch in the two species of Fritillaria L.was the characteristic B-type which was in consistent with that of potato starch. The degrees of crystallinity of F. thunbergii starch and F. ussurensis starch were about 29.9% and 20.1%, respectively. However,the degree of crystallinity of the potato starch was 44.9%. From the crystallinity degree of the starch in two species of Fritillaria L. , it could be concluded that the content of amylose in F. ussurensis starch was higher than that in F. thunbergii starch. The granule size of the starch in two species of Fritillaria L.ranged from 5 to 40 μm, which were all smaller than that of the potato. The starch granule in two species of Fritillaria L. was in cycloidal or elliptic-shape. It could be concluded that the thermal stability of the starch in two species of Fritillaria L. was different due to the different structures of different starch in various plants by TGA. Conclusion The physicochemical properties of starch in two different species of Fritillaria L. differ a lot due to their geographical origin.

Objective To carry out a systematic study on the chemical constituents in the fruits of Momordica grosvenori. Methods To isolate pure compounds by using repeated column chromatography,while the structure of a new compound was determined by detailed spectral analysis. Results Four cucurbitane triterpenoid glycosides, mogroside Ⅱ E (Ⅰ), mogroside Ⅲ (Ⅱ), grosmomoside Ⅰ (Ⅲ), and mogroside Ⅴ (Ⅳ) were isolated from the 50% ethanolic extract of the fruits of M. grosvenori. Conclusion Grosmomoside Ⅰ is a new compound identified as mogrol-3-O-β-D-glucopyranoside-24-O-{[β-D-glucopyranosyl (2-1 ) ]- [β-D-glucopyranosyl (6-1 ) ]-β-D-galactopyranoside } and the other three compounds are known compounds.

Objective To study the pharmacokinetics of aristolochic acid A in Radix Aristolochiae and the compound preparation of Guanxinsuhe Capsule in mice in vivo after single-dose oral administration and observe the difference of aristolochic acid A absorption and distribution. Methods Aristolochic acid A assay was performed by RP-HPLC on a Waters apparatus with a DiamonsilTM C18 column (250 mm × 4.6mm, 5 μm), a mobil phase: a mixture of methanol-water-acetic acid (72: 27 : 1), flow rate: 1.0 mL/min, detection wavelength: 315 nm, and column temperature: 20 ℃. Results Mice were given Radix Aristolochiae and Guanxinsuhe Capsule by ig at the same level of 2. 5 mg/kg of aristolochic acid A, respectively, which were suspended in 0. 3% CMC-Na solution. Plasma concentrations were determined by RPHPLC. After single-dose ig administration of Radix Aristolochiae or Guanxinsuhe Capsule to mice, the mean plasma concentration-time courses of aristolochic acid A obtained fitted the one-compartment model.The main pharmacokinetic parameters of aristolochic acid A in Radix Aristolochiae, t1/2ka, t1/2 ke, tmax,AUC, Cmax are 5. 103 min, 43. 63 min, 17.89 min, 80. 45 (μg · min)/mL, and 0. 916 8 μg/mL; the rela tive pharmacokinetic parameters in Guanxinsuhe Capsule are 5. 294 min, 43.50 min, 18. 32 min, 33.08(μg · min)/mL, and 0. 381 8 μg/mL. Conclusion The Cmax of aristolochic acid A in Guanxinsuhe Capsule is significantly less than that in Radix Aristolochiae, which indicates that the compound compability could decrease the absorption of aristolochiae acid A.

Objective To investigate the non-alkaloid constituent of Hippeastrun vittatum (Amaryllidaceae). Methods Solvent extraction and column chromatography were used to isolate the non-alkaloid constituents, and physicochemical constants and spectroscopic analysis were employed for structural elucidation. Results Five glycosphingosilipids were isolated, and their structures were elucidated to be (2S,3R, 4E, 8Z)-2-[(2R-2-hydroxyhexadecanoyl) amido ]-4, 8-octadecadiene-1, 3-diol 1-O-β-D-glucopyranoside ( Ⅰ ), (2S, 3R, 4E, 8E)-2-[(2R-2-hydroxyhexadecanoyl) amido]-4, 8-octadecadiene-1, 3-diol 1O-β-D-glucopyranoside ( Ⅱ ), (2S, 3R, 4E, 8Z)-2-[(2R-2-hydroxyoctadecanoyl) amido]-4, 8-octadecadiene-1, 3-diol 1-O-β-D-glucopyranoside (Ⅲ), (2S, 3R, 4E, 8E)-2-[(2R-2-hydroxyoctadecanoyl)amido]4, 8-octadecadiene-1, 3-diol 1-O-β-D-glucopyranoside ( Ⅳ ), (2S, 3R, 4E, 8Z)-2-[(2R-2-hydroxyeicosadecanoyl) amido]-4, 8-octadecadiene-1, 3-diol 1-O-β-D-glucopyranoside (Ⅴ), respectively. Conclusion They are all isolated from the fresh bulbs of H. vittatum for the first time.

Objective Application of a new molecular marker to the identification of Dendrobium (Orchidaceae) species. Methods Complete sequences of the mitochondrial nad 1 intron 2 for nine species of Dendrobium Sw. were amplified and determined. Results Seventeen variable sites were found in the aligned 872 bp of nad 1 intron 2 sequences. Eight of the nine Dendrobium species except D. loddigesii could be identified by the nad 1 intron 2 sequences. Conclusion The mitochondrial nad 1 intron 2 sequences could be used as a new molecular marker for the identification of Dendrobium species.

Objective To study the chemical constituents of the fruiting bodies of Hydnum repandum.Methods Separation and purification were performed on silica gel, Sephadex LH-20 and ODS CC. Their sturctures were established by 2D-NMR (1H-1HCOSY, HMQC, HMBC, and NOESY), MS, HR-MS spectra, and ORD data. Results Eleven compounds were obtained and identified as sarcodonin A ( Ⅰ ),scabronine B (Ⅱ), 3β-hydroxy-5α, 8α-epidioxyergosta-6, 22-dien (Ⅲ), (22E, 24R)-ergosta-7, 22-diene3β, 5α, 6β- triol (Ⅳ), (22E, 24R)-ergosta-7, 22-diene-3β-ol (Ⅴ), benzoic acid (Ⅵ), 4-hydroxylbenzaldehyde (Ⅶ), 4-monopropanoylbenzenediol (Ⅷ), ethyl-β-D-glucopyranoside (Ⅸ), thioacetic anhydride ( Ⅹ ), (2S, 2'R, 3S, 4R)-2-(2-hydroxyoctadecanoylamino) docosane-1, 3, 4-triol (Ⅺ). Conclusion All of the compounds are isolated from this fungus for the first time.

Objective To investigate the chemical costituents from Drymaria diandra. Methods Compounds were separated and purified by repeated column chromatographies on macroporous resin D101,silica gel, and RP-18.Two compounds were identified by spectral analysis. Results Two compounds were isolated from D. diandra. Theirs structures were identified as 6-carboxymethy1-5,7,4'-trihydroxyflavone (Ⅰ) and l-O-β-D-glucopyranosyl-(2S,3R,4E,8E)-Z-N-(2'-hydroxypalmitoyl) octadecasphinga-4,8-dienine(soya cerebroside Ⅰ,Ⅱ). Conclusion Compound I is a new compound. Compound Ⅱ is obtained from this plant for the frist time.

Object To investigate the chemical constituents from the roots of Glycyrrhiza uralensis Fisch. Methods The constituents were isolated on normal and reversed silica gel column chromatography and their structures were identified by spectral evidence. Results A new oleanane-type triterpenoid saponin and twelve known compounds, including two triterpenoid saponins, two cumarins and eight flavonoids, were isolated. Conclusion The new compound was elucidated as 3-O-[β-D-(6-methyl) glucuronopyranosyl (1→2)-D-glucuronopyranosyl]-24-hydroxy-glabrolide on the basis of ESI-MS, 1HNMR,13CNMR, HMQC and HMBC spectral evidence.

Object To explore the antitumor mechanism of Stellera chamaejasme Linn.(SC).Methods SC containing-serum(SCCS)was derived from mice pretreated with different doses of SC.Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used.Inhibition of proliferation was measured using MTT assay.Morphological assessment of apoptosis was performed with fluorescence microscope.DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry.Expression of bcl-2 protein was measured with immunohistochemistry.Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC(pretreated with SC3,6, and 12 g/kg)for 48h resulted in growth inhibition in a dose-dependent manner.Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced."Apobodies'in the apoptotic cells were observed,'ladder"pattern of agarose gel electrophoresis of DNA from 11.7% to 57.4%.Treatment with SC containing serum decreased the percentage of SGC-7901 cell of bcl-2 protein positive expression from 78.3% to 32.9%.Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.