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Objective To investigate the effect of effective fractions from Radix Vitis Davidii Foex. on fracture healing. Methods SD rats were randomly divided into model control group,Yunnan Baiyao group,and effective fraction group (n= 36 each group). Fracture models were established. Rats in the Yunnan Baiyao group and effective fraction group were given 3.0 g?kg-1?d-1 of Yunnan Baiyao and effective fraction from Radix Vitis Davidii Foex.,respectively,and rats in the model control group was treated with the same volume of pure water. Alkaline phosphatase and Ca concentration in serum were determined.For radius fracture structure,digital radiography and RT-PCR detection of VEGF mRNA expression were performed. Results The peak concentration of the alkaline phosphatase and Ca concentration in serum were significantly higher in the effective fraction group than in the Yunnan Baiyao group,and peak value appeared earlier,fracture healing score was higher, fracture line disappeared earlier,and VEGF mRNA expression was significantly higher in three weeks after the fracture (all P<0. 01) . Conclusion The effect of the effective fraction from RadixVitis Davidii Foex.on the fracture healing is strong.
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Objective To establish the quality standards system of A system to multiple evaluation on Yinxing huonao capusle,and discuss the feasibility of the method whether A system to multiple evaluation to be used for traditional Chinese medicine quality standard.Methods Identification of the Yinxing huonao capusle by TLC method was established by one thin layer chromatography system.HPLC was used to detect 6 types of the primary active components in Yinxing huonao capusle.Results 5 types of the primary active components were synchronize identified by one thin layer system.The color of spots was clear and had a good separation effect.6 kinds of the primary active components were synchronize detected by one system with high accuracy.Conclusion The established method A system to multiple evaluation can be used for the study of the quality standards system of Yinxing huonao capusle.
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Objective: To develop an HPLC-ELSD method for the simultaneous determination of six saponins constituents including notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd in Xinling pills.Methods: HPLC-ELSD was used, and the chromatographic separation was performed on an Agilent Eclipse XBD-C18 column (150 mm×4.6 mm,5 μm) with acetonitrile-water as the mobile phase with gradient elution at the flow rate of 1.0 ml·min-1, the column temperature was maintained at 20℃, the drift tube temperature was 60℃, and the gas pressure was 4.00 bar.Results: Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd was linear within the range of 0.30-6.00μg(r=0.999 5), 1.14-22.80μg (r=0.999 6), 0.17-3.40 μg (r=0.999 7), 0.81-16.20 μg (r=0.999 7), 0.08-1.60 μg (r=0.999 8) and 0.07-1.40 μg (r=0.999 8), respectively.The average recovery was 98.23%, 97.98%, 99.14%, 99.15%, 98.72% and 98.37%, and the RSDs were 1.56%, 1.31%, 1.16%, 1.07%, 0.73% and 0.92%(n=6), respectively.Conclusion: The method is convenient, accurate and reproducible in the quality control of saponins components in Xinling pills
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Objective To explore the extraction process of Yinxing Huonao capusle.Methods Central composite design-response surface method was used to optimize the extraction process with quantity of water and boiling time as main factors and with syringin transfer rate,emodin transfer rate and solids retention rate as evaluation index.Results The conditions of the optimized extraction process were as follows:10-fold water,extracting time of 160 min,extracting 2 times.Conclusion It is feasible that central composite design-response surface method is used to optimize the extraction process of Yinxing Huonao capusle,and the optimized extraction process is simple and reliable.
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OBJECTIVE:To establish the quality standard of Compound duzheng tablets. METHODS:TLC method was used for qualitative identification of Vitis davidii,Aralia chinensis,Acanthopanax senticosus and Panax notoginseng in the preparation. The contents of syringoside,oleanolic acid,ginsenoside Rg1,ginsenoside Rb1 and notoginsenoside R1 were determined by HPLC. The determination was performed on Inertsil ODS-SP C18 column with mobile phase consisted of acetonitrile-water(gradient elu-tion)at the flow rate of 1.0 mL/min. The detection wavelength was set at 215 nm,and the column temperature was 25 ℃. The sam-ple size was 10 μL. RESULTS:TLC spots of V. davidii,A. chinensis,A. senticosus and P. notoginseng were clear and well-separat-ed without interference from negative control. The linear ranges of syringoside,oleanolic acid,ginsenoside Rg1,ginsenoside Rb1 and notoginsenoside R1 were 0.056-0.337 μg(r=0.9998),0.022-0.130 μg(r=0.9999),0.030-0.182 μg(r=0.9995),0.010-0.061 μg(r=0.9998),0.038-0.226 μg(r=0.9999),respectively. RSDs of precision,stability and reproducibility tests were all lower than 3.0%. The recoveries were 100.55%-103.80%(RSD=2.0%,n=9),100.00%-104.56%(RSD=3.0%,n=9),97.06%-102.94%(RSD=2.1%,n=9)、97.06%-102.94%(RSD=2.0%,n=9),96.67%-100.00%(RSD=1.5%,n=9). CONCLUSIONS:Established standard can be used for the quality control of Compound duzheng tablets.
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SuperparamagneticSiO2@Fe3O4microspherewithcore-shellstructurewaspreparedbythe method of hydro-thermal and St?ber method, and modified by (3-aminopropyl) triethoxysilane ( APTES) . The obtained amino-Fe3 O4@SiO2 microsphere with controlled surface charge properties was characterized by SEM and TEM, and the surface electric property of amino-microsphere was investigated by Zeta potential measurement. The microsphere was used to extract DNA from human blood, and the solid phase extraction method by microspheres was developed. The mechanism of action between Aminated-Fe3 O4@SiO2 microsphere and DNA was explored, and the gel electrophoresis and PCR test were done on the extraction product. The result showed that genomic DNA with high purity was successfully extracted from whole blood by using amino-Fe3 O4@SiO2 microsphere, with the extraction rate about 70%, the saturated adsorption capacity for DNA was about 40 ng/μg, and the elution could be directly used for further bio-analysis.
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Based on the selective permeability of cationic exchange membrane and the characteristic of electri cal driven, a new and non-damage apparatus that can be used to separate compounds with different electric property was built to capture and enrich DNA.λ-DNA, as a model compound, was used to study the charac teristic of capturing and enriching DNA in this device.Factors that might affect the efficiency such as voltage and flow speed were optimized and gel electrophoresis was done to study if DNA was denaturation, PCR was done to prove the concentration can be used to further bioanalysis.Results were got; the apparatus could used to capture and enrich DNA scathelessly under the condition of low-voltage, the concentrate multiple was 62, and the recovery efficiency was accessible to 47%, concentration can be used for the next bioanalysis.Auto matic, easy to handle and little organic reagent using made it be advantageous, it could be miniatured to com bine with other biochips.This method had a large application space in field of DNA extraction, purification and concentration.
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The present study retrospectively analyzed 8 patients with hip dysplasia (8 hips) comprising 2 males and 6 females, aged 46 years old (range, 35-65 years old) who received treatment at the Department of Orthopedics, Second Affiliated Hospital of Nanchang University between September 2004 and November 2006. Preoperative radiographs and CT examination showed that all patients presented with sharp angle of affected hip > 45°, poor containment of femoral head, narrow joint space, and non-successive Shenton's line. Following Crowe classification, there were 5 cases (5 hips) of grade Ⅱ,and 3 cases (3 hips) of grade Ⅲ. Preoperative Harris hip score was 50 points (range, 44-62 points). Total hip arthroplasty was employed in each patient. Prosthesis was fixed by Cementless or cemented modes. Cementless fixation was applied in 6 hips and cemented fixation in 2 hips. Cementless prosthesis was used on the affected side in each patient. Replacement time, bleeding volume during surgery, hospital days post-surgery, Harris scores, and peri-operative complications were recorded.Simultaneously, post-surgery healing was observed. All 8 cases were followed up for an average period of 6 months. All patients exhibited well recovered joint function. No infection, sciatic nerve injury, dislocation of hip joint, fracture of femoral shaft, thrombosis of deep veins, or prosthesis loosening were found in any follow-up case. Six months after surgery, Harris hip score was 88 points (range, 86-94 points). Radiographs taken at 3 months post-surgery displayed the bone union of surgical region. All these findings suggest that acetabular medial wall displacement osteotomy in total hip arthroplasty produces obvious early- and middle-stage effects in treatment of hip dysplasia.
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Neural stem cells (NSCs) cultured in vitro were implanted within three-dimensional (3D) and self-assembly hydrogel from IKVAV-containing peptide. The cytocompatibility of hydrogel with NSCs was explored. Neural cells harvested from the cerebral cortex of neonatal mice were dissociated mechanically and cultivated in serum-free media, and were immunohistochemically examined for Nestin, NSE (neuron specific enolase) and GFAP (Glial fibrillary acidic protein); the self-assembly hydrogel from 1 wt% amphiphilic peptide was formed with the addition of DMEM/F12 and observed under transmission electron microscope (TEM); 1 x 10(5) /ml NSCs implanted within hydrogel (3D culture system) and seeded onto the surface of coverslips covered with polylysine (two-dimensional culture system, 2D culture system) respectively were incubated without serum at 37 degrees C in 5% CO2 and 95% air. Then the cells in 3D culture system and 2D one were double-labeled with DAPI/Brdu and observed under fluorescence microscope. The Nestin-positive cells were found, and they were able to differentiate into NSE-positive neuron- like cells and GFAP-positive glial-like cells, TEM showed that the hydrogel derived from 1 wt% peptide solution was composed of nanofiber network with the fiber diameters ranging from 3 to 6 nm and the fiber length warying from 100 nm to 1.5 microm, NSCs labeled with Brdu in 2D system were thinly scattered, and those in 3D system formed many neurospheres; the positive rate in 3D was much higher than that in 2D system (P < 0.001). The self-assembly hydrogel from the amphiphilic peptide containing IKVAV sequence had good cyto-compatibility and promoted the proliferation of NSCs.
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Animals , Rats , Animals, Newborn , Cells, Cultured , Cerebral Cortex , Cell Biology , Hydrogels , Pharmacology , Materials Testing , Neural Stem Cells , Cell Biology , Peptides , Chemistry , Rats, Sprague-Dawley , Surface-Active Agents , ChemistryABSTRACT
In this experimental study, the RGD-containing peptide was used to modify the surface of biomimetic PLGA-(ASP-PEG) matrix, and bone marrow stromal cells (BMSCs) were seeded onto these modified surfaces for three weeks. The effects of modified surfaces of matrix on the adhesion, proliferation and differentiation of BMSCs were explored. BMSCs were harvested from whole bone marrow of Sprague-Dawley (SD) rats in vitro, then were seeded onto peptide surface-modified matrix (Experiment group, EG) and matrices without modification (Control group, CG) respectively for three weeks. The number of adhesive cells was counted by using precipitation method after 4 h and 12 h incubation; the cells cytoskeletons were stained with FITC-conjugated phalloidin after 24h incubation; the cell density was investigated after 1 d, 2 d and 3 d of incubation; ALP activity of BMSCs was measured after 7 d, 14 d and 21 d of incubation with osteogenic medium. The cells from bone marrow were BMSCs and their purity was beyond 90% using flow cytometry (FCM) analysis. Sulphur binding energy in EG was shown by XPS to be 164 eV. BMSCs adhered on peptide surface-modified matrix were observed with SEM. Cell adhesion efficiency and quality in EG was better than that in CG, and cell cytoskeleton was more robust in EG. ALP activity was higher in EG than in CG. Peptide surface-modified PLGA-(ASP-PEG) was noted to have good compatibility with BMSCs and to promote cell adhesion and differentiation.
Subject(s)
Animals , Rats , Biocompatible Materials , Chemistry , Bone Marrow Cells , Cell Biology , Metabolism , Cell Adhesion , Cell Differentiation , Lactic Acid , Chemistry , Oligopeptides , Chemistry , Osteoblasts , Cell Biology , Peptides , Chemistry , Polyglycolic Acid , Chemistry , Rats, Sprague-Dawley , Stromal Cells , Cell Biology , Metabolism , Surface Properties , Tissue Engineering , Tissue ScaffoldsABSTRACT
45?,poor containment of femoral head,narrow joint space,and non-successive Shenton's line.Following Crowe classification,there were 5 cases(5 hips) of grade II,and 3 cases(3 hips) of grade III.Preoperative Harris hip score was 50 points(range,44-62 points).Total hip arthroplasty was employed in each patient.Prosthesis was fixed by Cementless or cemented modes.Cementless fixation was applied in 6 hips and cemented fixation in 2 hips.Cementless prosthesis was used on the affected side in each patient.Replacement time,bleeding volume during surgery,hospital days post-surgery,Harris scores,and peri-operative complications were recorded.Simultaneously,post-surgery healing was observed.All 8 cases were followed up for an average period of 6 months.All patients exhibited well recovered joint function.No infection,sciatic nerve injury,dislocation of hip joint,fracture of femoral shaft,thrombosis of deep veins,or prosthesis loosening were found in any follow-up case.Six months after surgery,Harris hip score was 88 points(range,86-94 points).Radiographs taken at 3 months post-surgery displayed the bone union of surgical region.All these findings suggest that acetabular medial wall displacement osteotomy in total hip arthroplasty produces obvious early-and middle-stage effects in treatment of hip dysplasia.