ABSTRACT
Objective:To establish an HPLC method for the content determination of cinacalcet hydrochloride and evaluate the un-certainty in the measurement. Methods:An Inertsil ODS-SP chromatographic column(250 mm×4.6 mm,5 μm) was used,the mo-bile phase was composed of phosphate buffer solution(pH 6.5) and acetonitrile(60: 40),the flow rate was 1.0 ml·min-1,the de-tection wavelength was 272 nm,the column temperature was 30℃ and the injection volume was 20 μl. The content calculation formula-tion was deduced,the influencing factors were determined,and each component was assessed. Results:The resolution between cina-calcet and the impurity was satisfied,the linear relationship within the range of 10-100 μg?ml-1was excellent(r=0.999 9),the av-erage recovery was 101.09% (RSD=0.54%,n=9),and the LOQ was 0.254 μg?ml-1. The expanded uncertainty was 1.22%, and the result of the content determination was(100.74 ± 1.22)% (k=2). Conclusion:The method is simple,fast,selective,ac-curate and reliable,and can provide reference for the development of quality standards for generic drugs. Based on the influencing fac-tors of uncertainty,the main influencing reasons for the determination results can be found out to improve the reliability of determina-tion.
ABSTRACT
Objective@#To observe effects of exogenous high mobility group protein box 1 (HMGB1) on angiogenesis in ischemic zone of early scald wounds of rats.@*Methods@#Thirty-six Sprague-Dawley rats were divided into HMGB1 group and simple scald (SS) group according to the random number table, with 18 rats in each group. Comb-like copper mould was placed on the back of rats for 20 s after being immersed in 100 ℃ hot water for 3 to 5 min to make three ischemic zones of wound. Immediately after scald, rats in HMGB1 group were subcutaneously injected with 0.4 μg HMGB1 and 0.1 mL phosphate buffer solution (PBS), and rats in SS group were subcutaneously injected with 0.1 mL PBS from boarders of ischemic zone of scald wound. At post scald hour (PSH) 24, 48, and 72, 6 rats in each group were collected. Protein expressions of vascular endothelial growth factor (VEGF) in ischemic zone of wound at PSH 24, 48, and 72 and protein expressions of CD31 in ischemic zone of wound at PSH 48 and 72 were detected by immunohistochemistry. The number of microvessel in CD31 immunohistochemical sections of ischemic zone of wound at PSH 48 and 72 was calculated after observing by the microscope. The mRNA expressions of VEGF and CD31 in ischemic zone of wound were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction at PSH 24, 48, and 72. Data were processed with analysis of variance of factorial design, t test, and Bonferroni correction.@*Results@#(1) At PSH 24, 48, and 72, protein expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=7.496, 4.437, 5.402, P<0.05 or P<0.01). At PSH 48 and 72, protein expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were 0.038 8±0.007 9 and 0.057 7±0.001 2 respectively, significantly higher than 0.013 4±0.004 9 and 0.030 3±0.004 0 of rats in SS group (t=10.257, 15.055, P<0.01). (2) At PSH 48 and 72, the number of microvessel in ischemic zone of wound of rats in HMGB1 group was obviously more than that of rats in SS group (t=3.536, 4.000, P<0.05). (3) At PSH 24, 48, and 72, mRNA expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.406, 3.821, 3.356, P<0.05). At PSH 24 and 48, mRNA expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.113, 3.466, P<0.05). At PSH 72, mRNA expressions of CD31 in ischemic zone of wound of rats in 2 groups were close (t=0.010, P>0.05).@*Conclusions@#Exogenous HMGB1 can promote angiogenesis in ischemic zone of early scald wounds of rats by increasing expressions of VEGF and CD31.
ABSTRACT
BACKGROUND:The decel ularized porcine smal intestinal submucosa is a kind of bioactive extracel ular matrix, which is mainly composed of col agen, glycoprotein, proteoglycan and rich in col agen, glycosaminoglycan and various growth factors, and these components play an important role in promoting the differentiation and proliferation of tissue cel s. OBJECTIVE:To prepare the injectable smal intestinal submucosa and to investigate its co-culture with rat adipose-derived mesenchymal stem cel s in vitro. METHODS:The injectable smal intestinal submucosa and rat adipose-derived stem cel s were prepared. Cel counting kit-8 test for cel proliferation:Passage 3 adipose-derived stem cel s were seeded onto the injectable smal intestinal submucosa (experimental group) and cel s cultured under normal condition as control group. The cel proliferation was observed at 1, 3, 5 and 7 days of incubation. Live/dead staining test for the survival of cel s:Passage 3 adipose-derived stem cel s were respectively cultured in the injectable smal intestinal submucosa extracts (experimental group) and complete culture medium (control group). Cel survival was determined at 1, 3, 5 and 7 days of culture. RESULTS AND CONCLUSION:Scanning electron microscope oval and strip adipose-derived stem cel s adhered onto the material. The absorbance values in the experimental group were higher than those in the control group at 1 and 5 days of incubation (P<0.05). Cel survival:The number of cel s appeared to be in a rising trend with time in both two groups;after 1-day co-culture, al cel s in the two groups survived. Then dead cel s appeared in both two groups, showing no significant difference. These results show that the injectable smal intestinal submucosa exhibits a good cytocompatibility.