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OBJECTIVE To stud y the chemical cons tituents of n-butanol part of Qubai tablet and its pharmacodynamic effect on the model of de melanocyte. METHODS The n-butanol part of Qubai tablet was prepared. The chemical constituents were analyzed by ultra-high performance liquid chromatography-mass spectrometry （UPLC-MS）. Taking mice B 16 melanoma cells as the research object ，the de melanocyte model was established and divided into model group ，positive control different concentration groups（8-methoxypsoralen 10，50，100，150，200 μmol/L），solvent group （diluted with DMSO ）and Qubai tablet n-butanol part different concentration groups （10，50，100，150，200 μmol/L）. The number of cells were observed by inverted microscope ，and the cell proliferation rate ，the rate of melanin production and promotion rate of tyrosinase activity were also detected. RESULTS In the positive and negative ion mode ，53 compounds in the n-butanol part of Qubai tablet were preliminarily determined （29 in the positive ion mode ，33 in the negative ion mode ，overlapping 9），of which coumarins accounted for the largest proportion ， followed by flavonoids. The n-butanol part of Qubai tablet could significantly increase the number of cells ，which was positively correlated with the action time and administration concentration. It could significantly increase the proliferation rate of cells ，the rate of melanin production and promotion rate of tyrosinase activity （P＜0.01）. CONCLUSIONS Coumarins and flavonoids may be the material basis for the anti-vitiligo effect of n-butanol part from Qubai tablet ；anti-vitiligo effect of n-butanol part of Qubai tablet may be realized by promoting tyrosinase activity.
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The pharmaceutical microbiological examination in medical institutions is an important method to ensure the safety of preparations.It is of great significance to popularize the techniques of drug microbiological examination in primary medical institutions.This article started with the purpose, plan and training contents of the microbiological examination training program, which combined with theory teaching and practical operation, two-phase practical teaching, group teaching and collective teaching and guided students to design and plan experiments to carry out the teaching work,and then explored the ways and improving measures of the pharmaceutical microbiological examination training for hospital pharmacists, which could provide reference for the training and teaching of hospital pharmaceutical microbiological examination.
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Objective:To establish an HPLC method for the simultaneous determination of triamcinolone acetonide acetonide ( TAA) and chloramphenicol in the cream. Methods:The chromatographic system consisted of a ZORBAX SB-C18 column (250 mm × 4. 6 mm, 5μm). The mobile phase consisted of methanol and phosphate buffer with gradient elution at a flow rate of 1. 0 ml·min-1, the detection wavelength was 240nm, the column temperature was ambient, and the sample size was 20 μl. Results:The calibration curve was linear for TAA and chloramphenicol within the concentration range of 6. 12-48. 96 μg·ml-1 and 62. 1-745. 2 μg·ml-1 with the recovery of 99. 7% (RSD=1. 3%, n=9) and 99. 4%(RSD=1. 0%, n=9), respectively. Conclusion: The method is sensitive, accurate and specific, and can be used to control the quality of chloramphenicol and triamcinolone acetonide acetate cream.
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OBJETCTIVE:To establish a method for the content determination of ethacridine lactate in Compound ethacridine ointment. METHODS:HPLC was performed on the column of Agilent ZORBAX SB-C18 with mobile phase of 0.1% Octanesulfon-ic acid sodium solution-acetonitrile(70∶30,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 270 nm,the column temperature was 30℃,and the injection volume was 10 μl. RESULTS:The linear range of ethacridine lactate was 10.002-50.010μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were less than 1%;recovery was 98.96%-100.36%(RSD=0.49%,n=9). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for content determina-tion of ethacridine lactate in Compound ethacridine ointment.
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Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of neomycin sulfate and hy-drochloric dyclonine in compound Twaln ointments. Methods:The assay was performed on an Agilent ZOR BAXSB-C18 column(250 mm × 4. 6 mm, 5μm) with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1. 0 ml·min-1 . The detection wavelength of DAD was 282 nm. The evaporator temperature of ELSD was set at 50℃ and the nebulizer temperature was set at 60℃with the gas flow rate of 1. 6 L·min-1 . The column temperature was kept at 35℃. Results:The linear range of neomycin sulfate was 141. 54-323. 52 μg·mL-1(r=0. 999 6) with the average recovery of 98. 87%(RSD=0. 95%, n=9). The linear range of hydro-chloric dyclonine was 28. 00-64. 00 μg·mL-1(r=0. 999 6) with the average recovery of 99. 57%(RSD=1. 10%, n=9). Conclu-sion:The method is accurate, sensitive and reproducible, and under the same chromatographic conditions, the determination of all the active ingredients in compound Twaln ointments is achieved, which provides basis for the quality control.
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Objective:To establish an HPLC method for the determination of chloramphenicol in chloramphenicol vaginal tablets. Methods:An Agilent ZORBAX SB-C18 column (150 mm × 4. 6 mm,5 μm)was used. The mobile phase was composed of 0. 1% sodi-um 1-heptanesulfonate solution-methanol (68∶32),the flow rate was 1. 0 ml·min-1, the UV detection wavelength was 277 nm,the column temperature was 35℃,and the injection volume was 10 μl. Results:The linear range of chloramphenicol was 25. 6-512. 0 μg ·ml-1(r=0. 999 9). The mean recovery was 99. 4%, and RSD was 0. 8%(n=9). Conclusion:The method is simple,accurate and reproducible,and can be used in the determination of chloramphenicol vaginal tablets.
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Objective To study the expression and the clinical significance of Wnt signal pathway regulation factor Pygo2 in human renal cell carcinoma.Methods Using RFQ-PCR and immunohistochemical SP methods to detect Pygo2 mRNA and protein expression in 42 cases renal clear cell carcinoma and their own normal kidney tissue specimens.All specimens had a definite diagnosis by pathologic.All were renal clear cell carcinoma.The tumor diameter was 1.2-15.5 cm.The average was 7.2 cm.Among all patients,there were 7 cases with diameter ＜ 4.0 cm,9 cases 4.0-7.0 cm,26 cases ＞ 7.0 cm.Fuhrman classification:grade Ⅰ 6 cases,grade Ⅱ 17 cases,grade Ⅲ 17 cases,grade Ⅳ 2 cases.AJCC TNM stages:stage Ⅰ 16 cases,stage Ⅱ 7 cases,stage Ⅲ 7 cases,stage Ⅳ 12 cases.Statistics was done to analyze the expression difference of pygo2 between normal kidney and renal cell carcinoma,and among renal cell carcinoma within each group.Results There was higher expression of Pygo2 in renal cell carcinoma,and in the adjacent lower expression.The pygo2 mRNA expression were 2.88 ± 1.26 and 1.00 ± 0.00 in respective specimens (P ＜ 0.0001).The pygo2 protein expression were 45.53 + 24.54 and 11.02 + 1.39 in respective specimens (P ＜ 0.0001).Pygo2 expression in grading and staging were statistically significant (P ＜0.0001),but in gender,age was not statistically significant (P ＞ 0.05).Conclusions High expression of Pygo2 was found in Fuhrman high grade,high clinical staging,lympho-metastasized renal clear cell carcinoma.Pygo2 might play an important role in the occurrence and development process in renal clear cell carcinoma.