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OBJECTIVE:To determine the content of phenolic acids from Anemone altica,and optimize its extraction technolo-gy. METHODS:HPLC was used to determine the contents of mono-ferulyl tartaric acid and ferulic acid from A. altica;using the total contents of 2 index components as index,volume fraction of extraction solvent,extraction solvent volume,extraction times and extraction time as factors,orthogonal test was used to optimize extraction technology,and verification test was conducted. RE-SULTS:The contents of mono-ferulyl tartaric acid and ferulic acid were 0.059%,0.0025%,respectively;the optimal extraction technology was as follow as 30% ethanol 600 mL added to 20 g medicinal material,extracted twice,90 min every time. In verifi-cation test,the average contents of 2 components in extract were 0.2971%(RSD=3.64%,n=3),0.0041%(RSD=5.11%,n=3). CONCLUSIONS:A method for contents determination of mono-ferulyl tartaric acid and ferulic acid from A. altica is estab-lished;optimized extraction technology is stable and feasible.
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Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative (FGFR1DN) on osteogenic induction culture of bone marrow stromal cells (BMSCs).Methods The 3rd generation BMSCs were divided into 4 equal groups (n =24).FGFR1-DN group was transfected by pcDNA3.1 (+)-FGFR1DN,FGFRI group by pcDNA3.1 (+)-FRFR1,blank load group by pcDNA3.1 (+)-blank vehicle and non-transtection group by nothing.After successful transfection was confirmed when the cells were in the logarithmic phase,osteogenic induction culture was conducted continuously for 21 days.Mineralized nodule formation was observed by alizarin red staining.The amount of mineralized material was calculated according to the standard curve of alizarin red concentration.Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining.The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by osteoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed diminished osteogenic capacity.The concentration of alizarin red was the highest in the FGFRI-DN group (1.33 ±0.19),the lowest in the FGFRI group (1.00 ± 1.17),and moderate in the blank load group (1.20 ± 0.16) and non-transfection group (1.17 ± 0.17),showing significant between-group differences (P ＜ O.05).Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase.This may provide a hint for local gene treatment of bone defects.
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Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative (FGFR1DN) on osteogenic induction culture of bone marrow stromal cells (BMSCs).Methods The 3rd generation BMSCs were divided into 4 equal groups (n =24).FGFR1-DN group was transfected by pcDNA3.1 (+)-FGFR1DN,FGFRI group by pcDNA3.1 (+)-FRFR1,blank load group by pcDNA3.1 (+)-blank vehicle and non-transtection group by nothing.After successful transfection was confirmed when the cells were in the logarithmic phase,osteogenic induction culture was conducted continuously for 21 days.Mineralized nodule formation was observed by alizarin red staining.The amount of mineralized material was calculated according to the standard curve of alizarin red concentration.Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining.The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by osteoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed diminished osteogenic capacity.The concentration of alizarin red was the highest in the FGFRI-DN group (1.33 ±0.19),the lowest in the FGFRI group (1.00 ± 1.17),and moderate in the blank load group (1.20 ± 0.16) and non-transfection group (1.17 ± 0.17),showing significant between-group differences (P ＜ O.05).Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase.This may provide a hint for local gene treatment of bone defects.
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Objective To explore the effect of fibroblast growth factor receptors 1-dominant negative strategy (FGFR1-DN) on alkaline phosphatase (ALP) activity of bone marrow stromal stem cells (BMSCs) after osteogenic induction.Methods BMSCs were transfected with eukaryotic expression plasmid pcDNA 3.1 (+)-DN FGFR1 and pcDNA3.1 (+)-FGFR1.The experiment was conducted in 4 groups:FGFR1-DN transfection group,FGFR1 transfection group,pcDNA3.1(+) empty vector transfection group and non-transfection group.The ALP activity of BMSCs was detected in logarithmic growth phase after osteogenic culture.The qualitative detection of ALP activity was carried out immunohistochemically while the quantitative detection by cALP kit.The ALP activity was compared between the 4 groups at 7 and 14 days after osteogenic induction.Results Compared with 7 days,the ALP activity at 14 days was significantly increased in the 4 groups,and the increase in FGFR1-DN transfection group was significantly higher than in the other 3 groups (P ＜ 0.05).At both 7 and 14 days,the ALP activity in FGFR1-DN transfection group was the highest while that in FGFR1 transfection group was the lowest (P ＜ 0.05).Conclusions FGFR1-DN can promote the ALP activity of BMSCs during osteogenesis.This may provide an experimental basis for the joint application of local gene therapy and tissue engineering and for construction of tissue engineered bone with better biocompatibility.
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BACKGROUND:There is little evidence on the use of al ogeneic tendon graft in the reconstruction of ankle joint. OBJECTIVE:To explore the clinical outcome of anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft in patients with chronic ankle instability. METHODS:Twenty-six patients with chronic lateral instability underwent anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon. There were 18 cases of simultaneous injury or chalasia in calcaneofibular ligament and anterior talofibular ligament, and 8 cases of anterior talofibular ligament injury or chalasia. The ankle joint function was evaluated according to AOFAS scale and Good classification. The affect ankle and healthy ankle were compared in the extension, plantar flexion activity, and metaleg activity. RESULTS AND CONCLUSION:Al the 26 patients were fol owed up for 9-24 months with a mean of 15 months. No cases appeared recurrent ankle lateral instability. The mean AOFAS score in the group of calcaneofibular ligament and anterior talofibular ligament was improved from (48.4±3.7) points preoperatively to (88.2±3.8) postoperatively, while that in the group of anterior talofibular ligament was improved from (50.0±6.4) points preoperatively to (89.5±3.4) points postoperatively. According to Good score, there were excellent in 19 feet, good in 6 feet, fair in 1 foot, with an excellent and good rate up to 96%. No serious complication was occurred in this group. Anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft can increase the tendon-bone contact area, improve the rate of tendon healing, and enhance the stability of ankle joint in patients with chronic ankle instability. Further studies are needed to verify its long-term efficacy.
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Objective To obtain differentiated osteoblast-specific inactivation of fgfr1 mice Methods To obtain fgfr1△/+/OC-CreTG/+ mice,fgfr1flox/flox mice obtained from NIH were crossed with OC-Cre mice To obtain fgfr1△/△/OC-CreTG/+ mutant mice,fgfr1△/+/OC-CreTG/+ further crossed with themselves or fgfr1flox/flox mice After fgfr1△/△/OC-CreTG/+ crossed with fgfr1flox/flox mice,half of their offspring were mutant mice Results Differentiated osteoblast-specific fgfr1 knockout mice were obtained Conclusion fgfr1△/△/OC-CreTG/+ mice were obtained through proper crossing strategy,which provides a suitable platform for studying fgfr1 function in bone development and fracture healing