ABSTRACT
Objective: To study the ectopic osteogenesis and biocompatibility of bone morphogenetic protein 2 (BMP-2)-derived peptide P24 loaded chitosan-4-thio-butylamidine (CS-TBA) hydrogel. Methods: First, the CS-TBA/hydroxyapatite (HA) solution was prepared by using chitosan, 2-iminothiolane hydrochloride, and HA. Then, the different amount of P24 peptides were added to the CS-TBA/HA to prepare the CS-TBA/5%P24/HA and CS-TBA/10%P24/HA solutions. Finally, β-glycerophosphate disodium (β-GP) was added to the CS-TBA/HA, CS-TBA/5%P24/HA, and CS-TBA/10%P24/HA to prepare the CS-TBA/HA/β-GP, CS-TBA/5%P24/HA/β-GP, and CS-TBA/10%P24/HA/β-GP hydrogels, respectively. Eighteen Sprague Dawley female rats were randomly divided into 3 groups ( n=6), which were injected into the back muscle pouches with equal volume CS-TBA/HA/β-GP hydrogel (group A), CS-TBA/5%P24/HA/β-GP hydrogel (group B), and CS-TBA/10%P24/HA/β-GP hydrogel (group C). The animals were sacrificed at 4 and 8 weeks and conducted micro-CT. The ability of biodegradation and osteogenesis of hydrogl was detected by trabecular thickness (Tb.Th), trabecular number (Tb.N), bone mineral density (BMD), and histological staining (HE and Masson). Results: All the rats in 3 groups survived to the time point of the harvest. Micro-CT results showed that the new bones gradually increased in each group after operation. At the same time, the new bone formation was more obvious in groups B and C than in group A, and with the increase of P24 concentration, new bone formation in group C was much more than that in group B. The Tb.Th, Tb.N, and BMD increased gradually in 3 groups, and the differences between 4 and 8 weeks were significant ( P<0.05) except the Tb.Th in group A. At different time points, the Tb.Th, Tb.N, and BMD were significantly higher in groups B and C than in group A ( P<0.05), and in group C was higher than in group B ( P<0.05), showing significant differences between groups. Histological staining showed that the materials of groups B and C were biodegradable, and the osteogenic effect was increased with the increase of P24 concentration. Conclusion: P24 peptide can improve the ectopic osteogenesis of CS-TBA hydrogel, and the 10% concentration is more effective.
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Objective:To establish an HPLC fingerprint and determine five compounds in Zhengtian pills to provide reference for the effective quality control.Methods:The analysis was carried out on an Agilent XDB-C18analytical column (250 mm × 4.6 mm,5 μm) with gradient elution by acetonitrile (A) -0.19% phosphoric acid solution (B) (0-8 min,10 % A→15% A;10-23 min,15 % A→45 % A;23-50 min,45 % A;50-60 min,45 % A→70 % A;60-70 min,70 % A),the detection wavelength was 310 nm and the flow rate was 1.0 ml/min. The column temperature was 30 ℃. Similarity evaluation was used to evaluate the fingerprints of 12 batches of Zhengtian pills,and five marker components were quantified. Results:There were 33 common peaks in the fingerprints of twelve batches of Zhengtian pills, and five of them (ferulic acid, prim-O-glucosylcimifugin, 5-O-methylvisammioside,imperatorin and isoimperatorin) were identified by comparison with the reference. The similarity of the 12 batches of samples was over 0.99. The linear ranges were 0.050-0.605 μg(r = 0.999 8),0.008-0.100 μg(r = 0.999 2), 0.013-0.150 μg(r = 0.998 6), 0.171- 2.049 μg(r = 0.999 7)and 0.113- 1.352 μg(r = 0.999 6) for ferulic acid, prim-O-glucosylcimifugin,5-O-methylvisammioside, imperatorin and isoimperatorin, respectively. The content of ferulic acid, prim-O-glucosylcimifugin,5-O-methylvisammioside,imperatorin and isoimperatorin was 0.78-0.84 mg/g,0.10-0.13 mg·g-1, 0.18-0.20 mg·g-1,2.44-2.51 mg·g-1and 1.70-1.78 mg·g-1in the 12 batches of samples, respectively. Conclusion:The established method has high sensitivity and specificity,which can be used for the quality control of Zhengtian pills.
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Objective:To optimize the extraction parameters for anthraquinones in Semen Cassiae by Box-Behnken experimental design. Methods:Based on single factor screening, a three-factor and three-level Box-Behnken experimental design was employed with the ethanol concentration, material to liquid ratio and extraction time as the independent variables. The dependent variables including the extration rates of chrysophand and aurantio-obtusin were transformed into desirability mathematically by Hassan 's method. The mathematical relationship was established between the dependent variables and the independent variables. The optimum experimental conditions were selected from the stationary point of the response surfaces. Results:The optimum extraction conditions were as follows:12-fold amount of 44％ ethanol as the solvent, extracted three times with 2. 65h for each time. The extraction rate of chrysophanol and aurantio-obtusin was 1. 921％ and 3. 244％, respectively. Conclusion:The measured value is close to the predicted one,which indi-cates the comprehensive extraction parameters optimized by Box-Behnken experimental design can be used for the extraction of chry-sophanol and aurantio-obtusin from Semen Cassiae.
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Objective To report therapeutic effects of patients with bladder cancer treated by intra-operative radiation and to recommend an effective method for the treatment of bladder cancer.Methods During the operation,when the tumor of bladder had been removed,a boost dose of electric beam from a linear accelerator was irridiated to the tumor bed.28 cases were follawed-up in a long period.Results Lower local recurrence was observed in patients received intra-operative radiation therapy,only 2 cases had a small recurrent tumor in the bladder during the follow-up period.26 cases had bean followed up,no tumor recurred.Conclusion The intraoperative radiation therapy is a useful treatment for the bladder cancer.