Evaluation of genetic diversity and population structure of genus Tripterygium based on SSR markers / 药学学报

The genus Tripterygium is an immune suppressor in the Chinese traditional medicines. Due to the habitat destruction and anthropogenic over-exploitation, the wild genus Tripterygium plants have decreased dramatically in recent years or even been endangered. It is critical to evaluate and protect genus Tripterygium wild resource. In this research, simple sequence repeat (SSR) molecular markers were applied to the investigation of the genetic diversity and genetic structure of 28 populations for genus Tripterygium (396 samples from 9 provinces in China). We found a high level of genetic diversity (percentage of polymorphic loci PPL=77.29%, Shannon's information index I=0.639 4; Nei's expected heterozygosity H=0.359 9) and high genetic differentiation among the populations (gene flow Nm=0.228 7). Based on Nei's genetic distance, the phylogenic tree of populations was constructed and 28 populations were divided into 6 clusters according to STRUCTURE clustering analysis. T. hypoglaucumwas was mainly divided into 3 clusters, including Sichuan, Yunnan and GuizhouChongqing. T. regelii was separated to cluster 4, while T. wilfordii was divided into two clusters:the transition type LQ and NY were divided into cluster 5, and the others were in cluster 6. These results provide a theory basis for the conservation of wild resource, research of genetic polymorphism and molecular marker for assisted breeding of genus Tripterygium.

Studies on difference of chemical compositions in plant species of Tripterygium genus / 中国中药杂志

We studied the content of chemical compositions and correlation among species of Tripterygium genus by principal component analysis(PCA) and variance analysis(ANOVA), and we also studied the difference among the 3 species.Using [BMIm]PF6 ionic liquid-based ultrasonic-assisted extraction, we determined the contents of 11 compounds including wilforgine, wilforzine, triptophenolide, wilforine, triptoquinone A, triptolide, tripterin, egallocatechin, epigallocatechin, catechin, and epicatechin in 28 batches of the Tripterygium species by HPLC and PCA. Partial least squares analysis (PLS) and ANOVA were also performed to verify the results.The analysis results of PCA and PLS showed that three species of Tripterygium genus were clustered into three regions respectively, and triptoquinone A was the important factor which affected the aggregation of these three species.There was a significant difference among the contents of 11 chemical components in the three species(P<0.000 1).These results indicated that there was a certain correlation between the chemical compositions and the classification of the species, and the difference of the chemical compositions among the three species was obvious. In this work, the content determination method is rapid and accurate, and the analysis method is simple and convenient, which provides a reference for the classification, the efficacy and the toxicity of the species.

Progress on molecular biology of Isaria farinosa, pathogen of host of Ophiocordyceps sinensis during the artificial culture / 中国中药杂志

Isaria farinosa is the pathogen of the host of Ophiocordyceps sinensis. The present research has analyzed the progress on the molecular biology according to the bibliometrics, the sequences (including the gene sequences) of I. farinosa in the NCBI. The results indicated that different country had published different number of the papers, and had landed different kinds and different number of the sequences (including the gene sequences). China had published the most number of the papers, and had landed the most number of the sequences (including the gene sequences). America had landed the most numbers of the function genes. The main content about the pathogen study was focus on the biological controlling. The main content about the molecular study concentrated on the phylogenies classification. In recent years some protease genes and chitinase genes had been researched. With the increase of the effect on the healthy of O. sinensis, and the whole sequence and more and more pharmacological activities of I. farinosa being made known to the public, the study on the molecular biology of the I. farinosa would be deeper and wider.

Species differences in caffeine metabolism in liver microsomes of rats and mice / 中国中药杂志

Caffeine and its metabolic products play an important role in clinical applications. An ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS/MS) method was applied to systemically study the caffeine metabolism in liver microsomes of rats and mice, and comprehensively evaluate caffeine metabolites in vitro and metabolism differences between species. The caffeine metabolites and metabolism differences between species in liver microsomes of rats and mice were analyzed by UPLC-Q-TOF-MS/MS high resolution mass spectrometry system and metabolitepolite software. The results showed that in addition to the demethylated and oxidized products in previous analysis, methylated, double oxidized, dehydrated and decarbonylated metabolites were also found in caffeine metabolism in liver microsomes of rats and mice, with significant difference in metabolism in vitro between rats and mice. The demethylated metabolite M2(C7H8N4O2) and decarbonylated metabolite M6(C7H10N4) in metabolism in vitro of mice were not found in rats, and the in vitro metabolite M7(C8H12N4O5) in rats were not found in mice. There was significant species difference in caffeine metabolism in vitro between rats and mice, providing important reference value for the further metabolism study and safety evaluation of caffeine.

Chemical constituents from Mylabris phalerata and their cytotoxic activity in vitro / 中国中药杂志

Ten compounds were isolated from Mylabris phalerata by using preparative HPLC and column chromatography over MCI gel. On the basis of physical-chemical properties, NMR and MS data analysis, the compounds were identified as 5'-[(1 R,2 R,3 S,6R)-1-hydroxymethyl-2-methyl-3,6-epoxycyclohexane-1,2-dicarboximide]- ethyl-2'-methyl-2'-butenoate (1),cantharidin (2), cyclo-(L-Pro-L-Ala) (3), cyclo-(R-Pro-R-Leu) (4), cyclo-(S-Pro-R-Leu) (5), cyclo-(D-Pro-L-Tyr) (6), indole-3-aldehyde (7), 3-indoleacetic acid (8), valerolactam (9), and 4-hydroxyphthalid (10).Compound 1 was a new compound, and compounds 2-10 were obtained from this genus for the first time. Compounds 1-9 were subjected to cytotoxic activity on HCT-116, HepG2, BGC-823, NCI-H1650, A2780 cell lines, and only compound 2 showed inhibitory effect on all cancer cell lines.

Proportion of Coptidis rhizoma and Evodiae fructus in the compound preparation: its effect in inducing apoptosis of SGC-7901 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of water extracts of Coptidis Rhizoma and Evodiae Fructus (CREF) on the proliferation and apoptosis of human gastric carcinoma cells (SGC-7901) and determine the optimal proportion of Coptidis rhizoma to Evodiae fructus.</p><p><b>METHODS</b>The growth inhibition of SGC-7901 cells treated with the water extracts of CREF of varying proportions was tested with MTT assay. The cell apoptotic rate and mitochondrial membrane potential were analyzed with flow cytometry.</p><p><b>RESULTS</b>The water extract of CREF with Coptidis Rhizoma: Evodiae Fructus proportions at 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1 all significantly inhibited the growth of SGC-7901 cells after a 24-h or 48-h treatment (P<0.05). The growth inhibition and cell death ratio both exhibited a dose-dependent pattern of Coptidis Rhizoma. Flow cytometry analysis showed that, after treatment of the cells with CREF at the proportions of 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1, the apoptotic rate were (8.50 ∓ 1.59)%, (9.90 ∓ 1.01)%, (17.15∓1.68)%, (21.55 ∓ 1.97)%, (34.10 ∓ 1.06)% and (34.40 ∓ 1.02)%, respectively, all significantly higher than that in the control group [(1.69 ∓ 1.91)%, P<0.05]. JC-1 Kit staining showed that mitochondrial membrane potential of SGC-7901 cells was decreased and the ratio of green to red fluorescence increased significantly after incubation with CREF.</p><p><b>CONCLUSION</b>CREF can inhibit the growth and induce apoptosis of SGC-7901 cells, and the strongest effect is achieved at the optimal proportion of Coptidis Rhizoma and Evodiae Fructus at 6:1.</p>

Determination of anthraquinone in Semen Cassiae from different regions by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the content of 7 anthraquinones in Semen Cassiae.</p><p><b>METHOD</b>A HPLC method was developed, with Inertsil ODS-3 column, acetonitrile and 0.1% H3PO4 solution as mobile phases in gradient elution. The detection wavelength wasset at 278 nm, and the flow rate was 0.8 mL x min(-1).</p><p><b>RESULT</b>Recoveries of all 7 anthraquinones were between 95%-105%. The content of the anthraquinones in crude drug produced in different habitation were different.</p><p><b>CONCLUSION</b>The method is convenient and accurate, which provides the foundation for the research of Semen Cassiae.</p>

Osmotically controlled oral drug delivery systems: an update review / 中国中药杂志

Osmotically controlled oral drug delivery systems (OCODDSs) utilize osmotic pressure for controlled delivery of active agents. The release of drugs from osmotic systems is governed by various formulations and processing factors such as solubility and pressure of the core components, properties of the semi-permeable membrane. In the present review, the references on OCODDSs have systematically been summarized in the following aspects: prescription design, industrial processing and equipments, methods for quality evaluation, and general situation of application. Prospect of applying the osmotic-pump technology into Chinese patent drugs is also discussed.

Effect of combination glycyrrhiizin and triptolide on TNF-alpha and IL-10 in serum of collagen induced arthritis rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effects of combination therapy with glycyrrhizin (GL) and triptolide (TP) on collagen-induced arthritis (CIA) rats.</p><p><b>METHOD</b>Sixty male SD rats were randomly divided into 6 groups: the model group, the TP group, the GL group, and combination 1, 2, 3 groups. The models were induced by collagen type II. The arthritis index (AI) and the edema rate were detected as curative effect, and the level of antibodies to collagen, TNF-alpha and IL-10 were measured by ELISA.</p><p><b>RESULT</b>The combination therapy with GL and TP significantly reduced the paw edema and arthritis index of CIA rats (P <0. 01 ), and the combination therapy can increase the level of IL-10, while decrease the level of TNF-alpha, and the level of antibodies to collagen decreased too (P <0.05, P <0.01).</p><p><b>CONCLUSION</b>Combine 26.78 mg x kg(-1) GL with 13.40 microg x kg(-1) TP can significantly inhibited the CIA, and the effect equal to the dosage of 17. 86 microg x kg(-1) TP. It supports the possible of GL in combination with TP to reduce the dose and side effects related to TP.</p>

Chemical constituents in roots of Polygala fallax and their anti-oxidation activities in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in roots of P. fallax and their anti-oxidation activities in vitro.</p><p><b>METHOD</b>Column chromatographic techniques were employed for isolation and purification of chemical constituents of the plant. The structures were elucidated on the basis of the spectral evidence and the physical and chemical character. The isolated compounds were screened with four anti-oxidation models in vitro.</p><p><b>RESULT</b>Seven xanthones, 1,7-dihydroxy-2,3-methylenedioxyxanthone (1), 1-methoxy-2,3-methylenedioxyxanthone (2), 3-hydroxy-1,2-dimethoxyxanthone (3), 1,6,7-trihydroxy-2,3-dimethoxyxanthone (4), 7-hydroxy-1-methoxy-2,3-methylenedioxyxanthone (5), 1,3-dihydroxy-2-methoxyxanthone (6) and 1,3,7-trihydroxy-2-methoxyxanthone (7), were isolated from the roots of P. fallax. And compounds 1 - 7 showed different anti-oxidation activities in the different pharmacological models.</p><p><b>CONCLUSION</b>Compounds 2, 3, 5 and 7 were isolated from this plant for the first time. Xanthones from this plant showed anti-oxidation activities. The pharmacological activities of the pure compounds from this plant were also reported for the first time.</p>

Studies on TLC fingerprint of flavonoids in rhizome of Polygonatum odoratum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the thin layer chromatographic (TLC) fingerprint of flavonoid constituents from Polygonatum odoratum, to set up the identification protocol of the herbal and provide scientific information for its quality control.</p><p><b>METHOD</b>The ethanol extracts were separated on silica gel G precoated plate with a mixture of toluene-ethylacetate-formic acid (5:4:1) as the mobile phase. The spots were visualized with ammonia vapor, then were examined under ultraviolet light (365 nm). The plate was scanned at wavelengths of lambdaR = 500 nm, lambdaS = 280 nm.</p><p><b>RESULT</b>A fingerprint of flavonoids of P. odoratum, with 10 specific fluorescent spots while examined under ultraviolet light, was set up.</p><p><b>CONCLUSION</b>The method can be used for quality control of P. odoratum.</p>