Research progress of studies on chemical constituents and biologic activities of Anemone species / 中国中药杂志

Anemone is an important genus which was distributed widely and used to folk medicines in China. It is rich of pentacyclic triterpenoid saponins,and more than 100 kinds of pentacyclic triterpenoid saponins had been isolated and identified. Anemone has been used to treat punch injury and rheumatoid arthritis. This article reviews the latest research progress of Anemone decoction from two aspects: chemical constituents and pharmacological. It will provide reference for further research and development of Anemone.

Research on purification and activity characterization of AmPR-10 ribonuclease from Astragalus membranaceus / 中草药

Objective To establish a stable and rapid separation and purification method for Astragalus membranaceus (Am) pathogenesis-related protein-10 (AmPR-10) using an automatic intelligent protein purification system AKTA Avant 25, and analyze its physiochemical and biological activity. Methods Am was extracted by Tris-HCl buffer. The crude extract was captured by anion exchange chromatography, and finely separated by hydrophobic chromatography and gel filtration chromatography. The relative molecular weight of AmPR-10 was measured by MALDI-TOF/TOF mass spectrometry, the protein identification was determined by mass spectrometry and MS/MS Ion Search, the glycoprotein identification was estimated by periodic acid-Schiff method, and the ribonuclease activity and effect factors were analyzed by agarose gel electrophoresis. Results The electrophoretically pure AmPR-10 was obtained by three-step purification of Q Sepharose Fast Flow, Butyl Sepharose High Performance and SuperdexTM 75 10/300 GL from the crude extraction. The relative molecular weight of AmPR-10 was 16 801. AmPR-10 was highly homologous to PR-10 and has no carbohydrate chains. Incubated at 56 ℃ for 30 min, AmPR-10 exhibited significant ribonuclease activity to total RNA of mammalian cells. The activity was insensitive to NaCl, pH value and mental ions, and weekly inhibited by 0.5 mol/L NaCl, pH 9.0, Mg2+ and Co2+. The activity was the same at EDTA as high as 20 mmol/L. Conclusion The three-step method of exchange chromatography-hydrophobic chromatography-gel filtration chromatography, a stable and rapid separation and purification method of AmPR-10, can be applied for other Chinese herbs. AmPR-10 might play an important role in resistance against virus.

Atributos físicos e químicos de agregados pedogênicos e de coprólitos de minhocas em diferentes classes de solos da Paraíba / Physical and chemical attributes of pedogenic aggregates and earthworm casts in different soil classes of Paraíba

A estrutura do solo é constituída de agregados pedogênicos e biogênicos, formados respectivamente pela hierarquização de agregados e pela ação da macrofauna, sobretudo a das minhocas. Neste estudo, objetivou-se a comparação entre agregados do solo (pedogênicos) e coprólitos de minhocas (biogênicos) de diferentes tamanhos e classes de solos. As coletas de amostras indeformadas de solos e coprólitos de minhocas (Pontoscolex corethrurus Muller, 1857) foram realizadas em quatro solos de classes diferentes do Estado da Paraíba (Latossolo Amarelo, Argissolo Vermelho Amarelo, Luvissolo Crômico e Nitossolo Vermelho). As caracterizações físicas e químicas de agregados e coprólitos foram realizadas após secagem à sombra e separação em três classes de diâmetros (20,0 - 9,52; 9,52 - 6,35 e 6,35 - 4,76 mm). A granulometria mostrou ser similar para cada tipo de agregado (pedogênico e biogênico), independentemente do tamanho dos agregados. Em geral, os coprólitos apresentaram maiores proporções de argila, silte e areia fina e menor proporção da fração areia grossa, maior estabilidade e maiores teores de carbono orgânico e cátions trocáveis, comparativamente aos agregados pedogênicos. Os resultados refletem um processo genético peculiar e sugerem a importância dos agregados biogênicos como indicadores da qualidade do solo.

The soil structure is composed by pedogenic and biogenic aggregates, formed respectively by aggregate hierarchy and macrofauna activity, especially by earthworms. The objective of this study was to compare soil aggregates and earthworm casts with different aggregate-size classes and soil classes. The sampling of undisturbed soil and earthworm casts (Pontoscolex corethrurus, Muller, 1857) was made in four soils of different classes in the State of Paraíba (Oxisol, Ultisol, Alfisol and Nitisol). The chemical and physical evaluations of aggregates and earthworm casts were determined after dry and split into three aggregate-size classes (20.0 to 9.52, 9.52 to 6.35 and 6.35 to 4.76 mm). The particle-size distribution was similar to aggregates and earthworm casts, independent of aggregate-size classes. Earthworm casts had higher proportions of clay, silt and fine sand, lower coarse sand, higher physical stability and more organic carbon and cations contents than the pedogenic aggregates. These results show a peculiar genetic process that makes biogenic aggregates an important indicator of soil quality.

The preparation of endostatin protein and the measurement of its biologic activity / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell.· METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.· RESULTS: pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts.·CONCLUSIONS: Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.

ADVANCES IN STUDIES OF SUBERGORGIC ACID / 中国海洋药物

Subergorgic acid is one of marine neurotoxins. It has a unique chemical structure. This article reviews its isolation and structural determination, total synthesis,derivatives preparation,biologic activities and potential applications.

The Study of the Preparation of Purified Fibronectin from the Human Plasma

Fibronectin(FN), a glycoprotein present in plasma and extracellular matrix, has been reported to be effective on various corneal disorders such as persistent epithelial defect, corneal trophic ulcer, herpetic keratitis, etc. We performed the study of the preparation of purified FN from 35 persons' plasma(male: 19, female: 16)by the use of Sepharose 48 Affinity Chromatography and Gel filtration. The prepared FN was pure electrophoretically, and no other plasma proteins were contaminated, and confirmed as pure by multiple methods such as SDS-P AGE, Ouchterlony double gel diffusion and immunoelectro phoresis. Only 40 minutes was taken for preparation. The result showed that FN concentrations in plasma and in prepared solution were 288.9 +/- 72.8 microgram/ml and 294.3 +/- 41.4 microgram/ml respectively. FN concentrations in plasma and FN eyedrops showed no difference between sex, but increased their level with age(p<0.05). The biological activity was better preserved at 4 degrees C(in the refrigerator) than at room temperature till about 4 weeks after preparation. Applicability of this method as a useful preparation of purified FN is recommendable.