OBJECTIVE To establish a method for detecting adenovirus type 7 by cells culture combined with real-time fluorescent RT-PCR.METHODS After purified adenovirus was dissociated from nasopharyngeal secretion in A549 cells,ADV7 E1A genes were detected by real-time RT-PCR assay and sequence analysis of cells infected with 0.1,0.5,5.0 and 10.0 MOI ADV7 at 3,6,12 and 24 h postinfection.Then the adenovirus in nasopharyngeal secretion was detected with the similar method.RESULTS Early transcription of E1A genes of adenovirus type 7 could be detected by real-time RT-PCR at 3 h postinfection with 0.5MOI virus;or at 6 h postinfection with 0.1MOI virus;Early transcription of E1A genes could be detected at 6 h postinfection in nasopharyngeal secretion.CONCLUSIONS The method by cells culture combined with real-time fluorescent RT-PCR is sensitive,specific and rapid.It can be applied in clinics for diagnosis of adenovirus type 7 infection.

Objective:To prepare the lithium-doped poly-glycerol sebacate (PGS-Li) scaffold using the specific effects of lithium ions and the excellent performance of PGS, and to provide the basis for its application prospects in cementation tissue engineering scaffold.Methods:The scaffolds were divided into two groups.The PGS-Li scaffolds prepared by adding lithium phosphate during the PGS cross-linking process were used as PGS-Li group, and the PGS scaffolds synthesized by the equal-purification of sebacic acid and glycerol were used as PGS group.The molecular weights of the scaffolds in two groups were determined by gel permeation chromatography.The structures of the scaffolds in two groups were analyzed by fourier transform infrared spectroscope.The surface morphology and the porosities and the pore sizes of the scaffolds in two groups were observed by scanning electron microscope.X-ray photoelectron (XPS) spectroscope and inductively coupled plasma optical emission spectrometer were used to determine the Li ion contents in the scaffolds in two groups.Thermogravimetric analyzer was used to analyze the thermal stabilities of the scaffolds in two groups.Contact angle measuring instrument was used to compare the hydrophilicities of the scaffolds in two groups.In vitro weight loss test was used to determine the degradation rates of the scaffolds in two groups.The OCCM-30cells were divided into experimental group (added with PGS-Li scaffold extract) , PGS group (added with PGS scaffold extract) and blank control group (added with DMEM culture medium) .MTT assay was used to detect the proliferation activities of cells in various groups at different time (24, 48and 72h) ;the cell morphology was observed by calcein-AM staining.Results:The gel permeation chromatography results showed that the molecular weight of the PGS-Li scaffold was slightly larger than that of the PGS scaffold.The specific absorption peak of phosphate was detected in the fourier infrared spectrum of the PGS-Li scaffold.The scaffolds in two groups had irregular three-dimensional network structures under scanning electron microscope, and the pore size was 20-160μm, the porosity of PGS scaffold was (53.92±2.18) ％, and the porosity of PGS-Li scaffold was (53.58±1.73) ％, there was no statistical difference between two groups (P＞0.05) .The XPS results showed that a peak appeared at 54.9eV in PGS-Li group, which coincided with the Li 1s binding energy, while the inductively coupled plasma emission spectrometer results showed that the Li ion content in the PGS-Li scaffold was 0.084％.The thermogravimetric analysis results showed that PGS-Li scaffolds began to degrade at a higher temperature and ceased at a lower temperature compared with PGS scaffolds.The contact angle measurement results indicated that both the materials were hydrophilic materials;the contact angle of PGS scaffold meterial was 78.26°±2.00°, and the contact angle of the PGS-Li scaffold material was 69.78°±1.15°;there was statistical difference between two groups (P＜0.05) .The in vitro degradation experiments showed that the degradation rate of PGS-Li scaffolds was faster than that of PGS scaffolds.The proliferation activity of OCCM-30cells in PGS-Li group had no significant difference compared with PGS group and blank control group (P＞0.05) .The calcein-AM staining results showed the green fluorescence in the OCCM-30cells in PGS and PGS-Li groups, and there were no significant changes in the morphology of cementoblasts.Conclusion:PGS-Li scaffolds have similar composition and structure to PGS scaffolds, and have better performance in hydrophilicity and thermal stability.PGS-Li scaffolds have no effect on the proliferation of cementoblasts and have broad application prospects in cementum tissue engineering.

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.