OBJECTIVE:To optimize the extraction technology of Compound qima capsules. METHODS:With the blood pres-sure lowering of rats as index,pharmacological efficacy test was used to screen the preparation technology(A was whole herb de-coction;B was Gastrodia elata fine powder mixed with other decocted medical materials). The extraction technology was opti-mized by single factor and orthogonal test using the contents of astragaloside and isoflavone grape glycosides and the quality of sol-id as indexes,with added water,decoction time,decoction times as factors;and the verification test was carried out. RESULTS:Pharmacological efficacy test showed that antihypertensive effect of sample by technology B was superior. The optimal extraction condition of other medical materials of technology B was as follows as 12-fold water per time,decocting for 1.5 h,for 3 times. In verification test,average extraction rates of astragaloside and isoflavone grape glycosides were 64.02% and 51.97%,and average value of the quality of solid was 5.69 g(RSD≤1.92%,n=3). CONCLUSIONS:The optimized extraction technology is stable and feasible.

BACKGROUND: Compared with bone marrow-derived mesenchymal stem cells (MSCs),human umbilical cord blood (hUCB)-MSCs possess many advantages,including easy acquirement,low immunogenicity,able to tolerance a higher degree of HLA-matching inconsistency,and with higher purity.OBJECTIVE: To analyze the method and condition for in vitro isolation,purification,proliferation,and osteoblast and lipoblast differentiation of hUCB-MSCs.DESIGN,TIME AND SETTING: The present observational experiment was performed in the Key National Laboratory of Biological Treatment,Huaxi Hospital,Sichuan University (i.e.,Institute of Stem Cells and Tissue EngIneering) between September 2004 and November 2005.MATERIALS: Neonatal cord blood at gestational age 37 to 40 weeks.METHODS: hUCB-MSCs were collected in aseptic condition,isolated by density gradient centrifugation,sedimenting red cells with methylcellulose followed by density gradient centrifugation,or immunomagnetic beads negative technique of CD34+.Theisolated MSCs were used to carry on plastic adherent culture in L-DMEM +10% fetal bovine serum (FBS) or MesencultTM medium +10% FBS.The third passage of cells were used for surface antigen determination by flow cytometry and induced to differentiate towards osteoblasts and lipoblasts.MAIN OUTCOME MEASURES: ①Identification of hUCB-MSCs.②Confirmation of hUCB-MSC differentiation by alizarin red and oil red staining.RESULTS: The mononuclear cells isolated by sedimented and centrifuged way cultured in MesencultTM medium +10% FBS were most available.Obvious colonies appeared in the third passage.The cells obtained by only centrifugation in density gradient were hard to form colony,and those isolated by immunomagnetic beads were hard to culture.The surface antigens of these colony cells presented CD29,CD59,and CD71,hut did not express CD34,CD45,and HLA-DR,and so on.After alizarin red staining,osteoblast-differentiating colony cell cytoplasm exhibited mineralized matrices.After old red staining,lipoblast-differentiating colony cell cytoplasm was full of lipid vacuoles.CONCLUSION: The MSCs can be successfully isolated by sedimenting red cells with methylcellulose followed by centrifugation and cultured in MesencultTM medium +10% FBS.Obvious colony growth appears in the third passage.After induction,the MSCs can differentiate into osteoblasts and lipoblasts.

OBJECTIVE:To study the purification technology of Sanhuang yishen formula. METHODS:Using retention rate and impurity rate of purified total polysaccharide,astragaloside and calycosin glucoside as index,the purification effects of water extraction and alcohol precipitation method (50%,60%,70% ethanol) and clarifying agent method (101 juice clarifying agent, ZTC natural clarifying agent,chitosan clarifying agent) were respectively detected to screen the purification method;orthogonal test was used to optimize the technology parameters(mass concentration of liquid,amount of clarifying agent and pH of liquid)by the optimized purification method,and the verification test was conducted. RESULTS:The purification was better when using chito-san as clarifying agent with comprehensive score of 98.62;the purified technology parameters were mass concentration of liquid 1 g/mL,1% chitosan solution amount of 2 mL/g,pH 5.1;the average value of retention rate and impurity rate of purified total poly-saccharide, astragaloside and calycosin glucoside in verification test were 79.56%(RSD=1.24%, n=3), 78.11%(RSD=0.97%,n=3),79.46%(RSD=1.03%,n=3)and 32.18%(RSD=1.16%,n=3),respectively. CONCLUSIONS:Using chito-san as clarifying agent shows good purification effect for Sanhuang yishen formula,which is simple. The optimized technology is stable and feasible.