ABSTRACT
The practice of general practice emphasizes that the general practitioner is the backbone, and the primary health institution is the main body, which solves the main health problems of the commu-nity and meets the basic health service needs of the residents. Rehabilitation medicine occupies a prominent position in the community health service, and its related knowledge and skills are the important part of the whole medical education. However, rehabilitation medicine has more teaching contents, less training time and out-of-date teaching model, which have made the effect of rehabilitation medicine teaching not good. In the rehabilitation medicine teaching for training general practitioners, by strengthening the concept of reha-bilitation, elaborating teaching content, optimizing teaching ward-round, and outstanding ability as well as reforming evaluation way, we try to exercise the students' clinical thinking and improve clinical skills, to cultivate outstanding qualified general medical practitioners for community rehabilitation ultimately.
ABSTRACT
The present paper is aimed to explore the biological osteoinductive activity of recombinant human bone morphogenetic protein 9 (rhBMP-9) by various biological technologies. In this study, we firstly obtained hBMP-9 cDNA by PCR and inserted it into vector pcDNA4/His Max to reconstruct hBMP-9 eukaryotic expression vector pcDNA4/His Max-BMP-9. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level rhBMP-9 was reconstructed by co-transfecting the expression vectors pcDNA4/His* Max-hBMP-9 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification by the methotrexate. We finally obtained a monoclonal cell line expressing the highest level protein. We purified the medium after culturing the highest-producing monoclonal by Ni-NTA His-Bind Resin columns and concentrated to by a Centricon 50 at 4 degrees C and stored at 70 degrees C until it was used. Western blot and SDS-PAGE analyses showed a specific band of about 32kD in pro-region lane and a specific band of about 50kD in pro-region complex lane. Biological activities of rhBMP-9 were tested by colorimetric determination and histochemical staining of Alkaline Phosphatase (ALP) Activity, osteocalcin and oesteopontin for C3H10 T1/2 cells, which were stimulated culture by different concentration (20, 50, 100 microg/mL) of rhBMP-9. The results showed that the rhBMP-9 could induce osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro, and were proportional to the amount. This study can provide experimental data for further tests in vivo and clinical applications.
Subject(s)
Animals , Cricetinae , Humans , CHO Cells , Cricetulus , Growth Differentiation Factor 2 , Pharmacology , Osteogenesis , Recombinant Proteins , Pharmacology , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To establish quality control criteria for medicinal herb Cajanus cajan based on the determination of longistylin A and longistylin C, two bioactive and specific stilbenes of the plant.</p><p><b>METHOD</b>Longistylin A and longistylin C were obtained from the leaves of C. cajan by silica gel column chromatography and identified as marker compounds of this plant by spectroscopic analysis. A RP-HPLC method was established to determine the two compounds.</p><p><b>RESULT</b>Longistylin A and longistylin C were well separated on a Thermo BDS Hypersil C18 column (4.6 mm x 250 mm, 5 microm) with a mobile phase methanol-water (8:2), and showed good linearity in the range of 0.00288 - 0.0576 microg and 0.0112 - 0.224 microg, respectively. The average recoveries were 98.9% and 97.2% with RSD of 2.4% and 2.2% for these two compounds, respectively.</p><p><b>CONCLUSION</b>The established analysis method is simple and accurate, whicn can be used for quality control of C. cajan.</p>