For the purpose to seek the active components in the aerial parts of Verbena officinalis L.with anti-early pregnancy activity, the methanolic extract of this drug was repeatedly chromatographed onsilica gel column. Three iridoid glucosides were isolated and identified on the basis of physical, chemicalevidences and spectroscopic analysis (UV, IR, 1HNMR, 13CNMR, 1H-1HCOSY, 1H-13CCOSY and DEPT)as verbenalin (Ⅰ), 3, 4-dihydroverbenalin (Ⅲ) and hastatoside (Ⅲ).Ⅱ is a new natural compound.

Objective To study the effect of extracts of Smilax china L. on inhibition the experimentally induced benign prostatic hyperplasia ( BPH) , and screen the effective fraction. Methods The BPH model was built on the castrated rats by subcutaneous injection of testosterone propionate. Male rats were randomly divided into eight groups ( n=6 ):sham operation, model control, petroleum ether fraction, acetic ether fraction, n-butyl alcohol fraction, water fraction, macroporous resin fraction ( FMR) , and total extracts group. The rats were treated with testosterone propionate by subcutaneous injection for consecutive 3 weeks. Meanwhile, rats were orally administrated with the six extract fractions of S. china L. After the last administration, serum was separated for the determination of prostatic acid phosphatase ( PACP ) , prostate was weighed and histopathological examination was carried out to evaluate the inhibitory effect of S. china L. against BPH. Results All of the six fractions from S. china L. could inhibit BPH, and the n-butanol fraction, water fraction and FMR showed better inhibitory effect, which significantly decreased the prostatic index by 52. 80%, 50. 93% and 67. 70%, respectively, remarkably reduced serum PACP, and notably improved the prostate gland morphology compared with the model group. Among the three fractions, FMR showed the strongest effect against BPH. Conclusion S. china L. ameliorates the experimentally prostatic hyperplasia, and FMR showes the best effect, which might be the bioactive components against BPH.

A simple method has been proposed for the determination of clozapine (CLZ) and chlorpromazine (CPZ) in human urine by dispersive liquid-liquid microextraction (DLLME) in combination with high-performance liquid chromatography-ultraviolet detector (HPLC-UV). All important variables influencing the extraction efficiency, such as pH, types of the extraction solvent and the disperser solvent and their volume, ionic strength and centrifugation time were investigated and optimized. Under the optimal conditions, the limit of detection (LODs) and quantification (LOQs) of the method were 13 and 39 ng/mL for CLZ, and 2 and 6 ng/mL for CPZ, respectively. The relative standard deviations (RSDs) of the targets were less than 5.1% (C=0.100 μg/mL, n=9). Good linear behaviors over the tested concentration ranges were obtained with the values of R (2)>0.999 for the targets. The absolute extraction efficiencies of CLZ and CPZ from the spiked blank urine samples were 98.3% and 97.8%, respectively. The applicability of the technique was validated by analyzing urine samples and the mean recoveries for spiked urine samples ranged from 93.3% to 105.0%. The method was successfully applied for the determination of CLZ and CPZ in real human urine.

Objective To study the stilbene constituents from the traditional Chinese medicine Smilax china and to determine their antioxidant activity. Methods The compounds were separated and purified by column chromatography with silica gel, RP C18, and Sephadex LH- 20, and were identified by IR, MS, NMR. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. Results Three compounds were isolated from the EtOAc fraction of the rhizomes of S. china and were identified as: resveratrol (1), oxyresveratrol (2) and 3, 5, 3′ , 4′ - tetrahydroxylstilbene (3). Compounds 1～ 3 showed strong antioxidant activity, and could scavenge DPPH free radicals, effectively. At the concentration of 50 ? mol/L, their DPPH free radical scavenging rates were 79.47 % , 89.89 % and 93.86 % , respectively. Conclusion Stilbenes might be the material foundation of antioxidant activities of rhizomes of S. china.

Objective:To explore the effect of the methanol extract of Macrothelypteris oligophlebia on chronic non-bacterial prosta-titis ( CNP) in rats to confirm the active fractions. Methods:The powdered rhizomes of M. oligophlebia were soaked in methanol. The methanol extract was suspended in water and then extracted successively with chloroform and ethyl acetate to obtain chloroform fraction, ethyl acetate fraction and water fraction. Carrageenan-induced CNP in rats was established. The rats were randomly divided into the sham-operated control group, model group, positive control group, methanol extract group, ethyl acetate fraction group, chloroform fraction group and water fraction group. The anti-prostatitis effect was evaluated by the prostate index, and the pathological examination of prostate was performed using HE staining. The levels of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-α), cyclooxyge-nase-2 (COX-2) and prostaglandin E2(PGE2) were analyzed using ELISA kits. Results:The ethyl acetate fraction group and metha-nol extract group with high flavonoid content could significantly decrease prostate index (P<0. 01) and the levels of IL-10, TNF-α, COX-2 and PGE2(P<0. 05 or P<0. 01), and improve the prostate morphology when compared with the model group, especially with the ethyl acetate fraction group. Conclusion:The rhizomes of M. oligophlebia show promising therapeutic effect on CNP, and the ethyl acetate fraction is the active fraction.

Objective To study the chemical constituents of Abacopteris penangiana.Methods The compounds were separated and purified by various chromatographic techniques and their structures were elucidated on the basis of physiochemical properties and spectroscopic methods.Results Seven compounds were purified and their structures were identified as:(7'Z)-3-O-(3,4-dihydroxy phenylethenyl)-caffeic acid(1),caffeiein B(2),matteucinol(3),protocatechuic acid(4),p-methoxybenzoic acid(5),β-sitosterol(6),and daucosterol(7).Conclusion Compound 1 is a new phenolic acid compound named abacopteric acid,and the other compounds are isolated from the plant for the first time.

OBJECTIVETo investigate the effect of antisense vascular endothelial growth factor (VEGF)(121) cDNA transfection on the growth of K562 cells in nude mice.

METHODSK562 cells transfected with the antisense (AS) or sense (S) VEGF(121) cDNA, and the vector (V, pcDNA3) alone were transplanted subcutaneously into nude mice and the growth of the transfected cells in vivo was investigated. The effects of transfected K562 cells on human bone marrow endothelial cells (BMEC) were analyzed by MTT assay, the microvessel density (MVD) in tumor mass by vWF immunohistochemistry stain.

RESULTSK562/V tumor grew more slowly [(207.5 +/- 192.9) mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.05] and K562/S tumor more rapidly than K562/V tumor did [(1 174.6 +/- 508.7)/mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.01]. K562/S cell culture supernatant was more strongly in promoting the proliferation of BMEC than K562/V supernatant did, but K562/AS supernatant resulted in a marked decrease of the promoting effect as compared with K562/V's. The MVDs in K562/AS, K562/S, and K562/V tumors were [(11.0 +/- 7.6)/0.72 mm(2) vs (50.8 +/- 11.7)/0.72 mm(2) vs (18.9 +/- 7.0)/0.72 mm(2)], respectively.

CONCLUSIONSAntisense VEGF(121) cDNA transfected K562 cells show growth retardation in transplanted nude mice, decrease of tumor MVD, and decrease of promoting BMEC proliferation capacity.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Division ; genetics ; physiology ; Culture Media, Conditioned ; pharmacology ; DNA, Antisense ; genetics ; DNA, Complementary ; genetics ; Endothelial Growth Factors ; genetics ; physiology ; Endothelium, Vascular ; cytology ; drug effects ; Female ; Humans ; K562 Cells ; Lymphokines ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; blood supply ; genetics ; pathology ; Neovascularization, Pathologic ; genetics ; physiopathology ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

According to our previous experiments, Ph(+) chronic myeloid leukemia (CML) cell line K562 cells have defects in beta 1 integrin activation. In order to search the same regularity in Ph(+) CML bone marrow cells, bone marrow mononuclear cells (BMMNC) from 12 cases of Ph(+) CML and 10 cases of normal individuals were studied. Their expression rate of 9EG7 epitope on beta1 integrin post treatment by 8A2 or GM- or G-CSF and cell adhesion ability with soluble fibronectin (FN) were evaluated by flow cytometry; in addition, the effects of CGP57148B, a highly specific ABL tyrosine kinase inhibitor, were observed. Our results showed that 9EG7 expression rate and FN binding rate were very low in all the inactivated cells. The parameter increased markedly post 8A2 activation in both NBMMNCs and CMLBMMNCs, but the degree of increase in CMLBMMNCs was significantly lower than that in NBMMNCs; GM-CSF or G-CSF could significantly increase the parameters in NBMMNCs while had no effects on that in CMLBMMNCs. CGP57148B could increase the beta1 integrin activation potential of CMLBMMNCs but had no effects on that of NBMMNCs. The results indicate that decreased activation potential of beta1 integrin in CMLBMMNCs is the major cause of adhesion defects of Ph(+) CML cells; beta1 integrin functional insufficiency in CMLBMMNCs could not be directly reversed by ABL tyrosine kinase inhibitor CGP57148B.

OBJECTIVETo study the chemical constituents of Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell.

METHODThe compounds were isolated by column chromatography with silica gel, C18 reverse-phase silica gel, sephadex LH-20, and their structures were elucidated on the basis of physiochemical propertities and spectral analysis. The antitumor activities of all compounds were tested with MOLT4, Hep G2, A-549, MCF-7, HT-29, PC-3 tumor cell lines.

RESULTFive compounds were isolated and identified as protoapigenone (1), protoapigenin (2), protoapigenin-4'-O-beta-D-glucopyanoside (3), 5,7-dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl) -chromen-4-one (4), 5,7-dihydroxy-2-(1-hydroxy-2,6-dimethoxy-cyclohex-4-oxo) -chromen-4-one (5), respectively.

CONCLUSIONAll compounds were obtained from this plant for the first time. Compounds 1, 4 and 5 showed strong anti-proliferative effects on six tumor cells, which were in concentration-dependent manner.

Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ferns ; chemistry ; drug effects ; Flavonoids ; chemistry ; pharmacology ; HT29 Cells ; Hep G2 Cells ; Humans

A simple method has been proposed for the determination of clozapine (CLZ) and chlorpromazine (CPZ) in human urine by dispersive liquid-liquid microextraction (DLLME) in combination with high-performance liquid chromatography-ultraviolet detector (HPLC-UV).All important variables influencing the extraction efficiency,such as pH,types of the extraction solvent and the disperser solvent and their volume,ionic strength and centrifugation time were investigated and optimized.Under the optimal conditions,the limit of detection (LODs) and quantification (LOQs) of the method were 13 and 39 ng/mL for CLZ,and 2 and 6 ng/mL for CPZ,respectively The relative standard deviations (RSDs) of the targets were less than 5.1％ (C=0.100 μg/mL,n=9).Good linear behaviors over the tested concentration ranges were obtained with the values of R2＞0.999 for the targets.The absolute extraction efficiencies of CLZ and CPZ from the spiked blank urine samples were 98.3％ and 97.8％,respectively.The applicability of the technique was validated by analyzing urine samples and the mean recoveries for spiked urine samples ranged from 93.3％ to 105.0％.The method was successfully applied for the determination of CLZ and CPZ in real human urine.