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ObjectiveTo establish a rapid screening method for influenza virus neuraminidase（NA） inhibitors sourced from Chinese medicines based on fluorescence detection. MethodThe method was constructed based on the principle that after the reaction of the test sample and a certain amount of NA， the activity of some NA will be inhibited by the test sample， and the NA that is still active after the addition of the substrate can generate fluorescence at a specific wavelength when combined with the fluorescent substrate， and the inhibition rate of the test sample on NA was calculated according to the measured fluorescence intensity， so as to evaluate the in vitro inhibitory activity of the test sample on NA. A total of 49 high-purity chemical components from 12 Chinese medicines were used to evaluate the in vitro anti-NA activity by the established method. The theoretical calculated values of binding energy and inhibition constant after docking between the NA protein receptor and the test sample were used to prove the accuracy of the experimental results. The established method was applied to detect the in vitro NA inhibitory activity of different batches of Banlangen granules and Kangbingdu granules， so as to evaluate the quality consistency among different batches of samples. ResultThe methodological examination results showed that the method had good accuracy and repeatability. The screening results of 49 components showed that 22 of them had strong in vitro inhibitory activity against NA than peramivir ［half inhibitory concentration（IC50） was 131.2 μmol·L-1］， such as schaftoside， isoorientin， chebulinic acid， menthone and isoschaftoside. The inhibitory activity of the remaining 27 components was weaker than that of peramivir. The molecular docking results showed that the theoretical calculation results of binding energies and inhibition constants of most compounds were basically consistent with the experimental results. The test results of the inhibitory activity of 12 batches of Banlangen granules on NA showed that the quality consistency among samples A1， A2， B2， C1， C2， E2 and F2 was good. The analysis results of the inhibitory activity of 9 batches of Kangbingdu granules produced by the same manufacturer on NA showed that the inhibitory rates of samples K1 to K9 were 37.68%， 36.18%， 31.37%， 33.98%， 40.36%， 33.76%， 40.69%， 41.08%， 40.06% when the concentration of 0.02 g·mL-1， and the average inhibitory rate was 37.24%. ConclusionIn this paper， we successfully established an analytical method that can be used to rapidly evaluate whether Chinese medicines （derived from chemical components of traditional Chinese medicine or proprietary Chinese medicines） have in vitro anti-NA activity， which can be a powerful supplement to the existing screening methods for influenza virus NA inhibitors. And this method was used to screen 22 compounds from 12 Chinese medicines with good in vitro inhibitory activity against NA， which can provide candidate compounds for the development of anti-influenza small molecule drugs.
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Objective To establish HPLC method for determination of (R,S)-epigoitrin in Banlangen granules and discuss the content limitation. Methods The samples were separated on SHIMADZU VP-ODS (150 mm×4. 6 mm, 5 μm) and the mobile phase consisted of methol-0. 02% phosphoric acid solution (7:93) at a flow rate of 1. 0 mL. min-1 . The detection wavelength was set at 245 nm and the injection volume (in automatic sampler) was 10 μL. The content limitation was assessed according to the transfer rate and statistical data of the results. Results (R,S)-epigoitrin showed a good linear relationship at concentration of 0. 058 7 – 150. 349 5 μg·mL-1(r = 0. 999 7, n = 7). The average recovery rate was 98. 72% , 98. 40% and 98. 60% , respectively; RSD was 1. 84% , 0. 50% and 1. 82% , respectively. The content limitation of (R,S)-epigoitrin was unreasonable according to the transfer rate and the statistical data of the results. Conclusion The method is easy and simple to operate, accurate and stable in results, and highly specific, thus it is applicable for the quality control of Banlangen granules. The content limitation should be determined on the basis of pharmacokinetic and pharmacodynamic data.
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Objective:To observe the clinical curative effect of integrated traditional Chinese and western medicine in the treat-ment of patients with ocular herpes zoster. Methods:Seventy patients with ocular herpes zoster in our hospital were selected during De-cember 2011 to December 2013. According to random number table,the patients were divided into two groups. The patients in the con-trol group were treated with ganciclovir 5. 0 mg·kg-1 ·h-1 , ivd, qd and ganciclovir ophthalmic gel, qid. The patients in the observa-tion group were treated with Banlangen granules and acupuncture additionally. The clinical efficacy, the time of blistering, pain and crusting and adverse reactions in the two groups were observed and compared. Results:After the treatment, the effective rate of the ob-servation group was 94. 29% , while that of the control group was 77. 14% (P0. 05). Conclusion:The clinical efficacy of integrated traditional Chinese and western medicine in the treatment of ocular herpes zoster is obviously better than that of western medicine treat-ment alone with fewer adverse reactions and shorter time of blistering, pain and crusting.
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OBJECTIVE:To establish an HPLC method for the determination of the content of Indirubin in Banlangen granules. METHODS: The choromatographic separation was performed on C18 column (250 mm?4.6 mm,5 ?m) with column temperature set at 30 ℃. The mobile phase consisted of methanol-water(76∶24) at a flow rate of 1.0 mL?min-1.The dete-ction wavelength was set at 289 nm.RESULTS:The calibration curve of Indirubin was linear in the range of 0.073 5～0.612 5 ?g (r=0.999 9) and the average recovery was 98.78%(RSD=0.74%,n=9).CONCLUISON: The method is rapid and accurate, and it can be used for quality control of Banlangen granules.