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ObjectiveTo explore the meridian tropism of components in Bupleuri Radix （Chaihu， CH） based on the model of nonalcoholic steatohepatitis （NASH） and clarify the substance basis of the meridian tropism of CH in Xiaoyaosan （XYS） by means of principal component analysis. MethodEighty SPF male C57BL/6 mice were randomly assigned into 8 groups， with 10 mice in each group. Except that the blank group was fed with the methionine choline-sufficient （MCS） diet， the other mice were fed with methionine choline-deficient （MCD） diet for 4 weeks to establish the nonalcoholic steatohepatitis （NASH） model. After the established model was confirmed by hematoxylin-eosin （HE） staining， the mice were administrated with corresponding drugs by gavage once a day for 4 weeks. Specifically， the 8 groups were XYS group （2.874 g·kg-1）， XYS-CH group （2.445 g·kg-1）， XYS-CH+volatile oils （Vol， 0.163 mg·kg-1） group， XYS-CH+polysaccharides （Pol， 24.067 mg·kg-1） group， XYS-CH+flavones （Fla， 2.241 mg·kg-1） group， and XYS-CH+saponins （Sap， 2.746 mg·kg-1） group. The model group and the blank group were administrated with the same volume of normal saline. After the last administration， the mice were sacrificed for the collection of blood and liver tissue. The pathological changes of liver were observed by HE staining and oil red O staining. Enzyme linked immunosorbent assay （ELISA） kits were used to determine the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein （HDL）， and low-density lipoprotein （LDL） in serum as well as malondialdehyde （MDA）， superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） in liver. SPSS Statistics 23 was used for principal component analysis and comprehensive evaluation to determine the substance basis of the meridian tropism of CH in NASH mice. ResultCompared with the blank control group， the modeling led to hepatocyte swelling， increased fat vacuoles， and appearance of inflammatory cells. Further， the modeling elevated the levels of ALT， AST， TG， TC， and LDL and lowered the HDL level in serum， and it increased the MDA level and decreased the SOD， CAT， and GSH-Px levels in liver. Compared with the model group， the administration of XYS and XYS-CH in combination with the components of CH alleviated the oxidative damage in liver （P<0.05）. The comprehensive score of the pharmacological efficacy was in a descending order as follows： XYS > XYS-CH+Sap > XYS-CH+Fla > XYS-CH+Pol > XYS-CH+Vol > XYS-CH. Among the chemical components of CH， Sap had the best effect. ConclusionSap lowers the blood lipid level， regulates the abnormal lipid metabolism， and alleviates the oxidative damage of liver， which is the substance basis for CH to exert the meridian tropism in liver.
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OBJECTIV E To establish the method for evaluating the quality o f Plantago asi atica and fried P. asiatica . METHODS The fingerprints of 15 batches of P. asiatica and 15 batches of fried P. asiatica were established by HPLC. The common peaks were identified with the Similarity Evaluation System for Chromatographic Fingerprinting of TCM （2012 edition）， and similarity evaluation was performed. Analysis of chemical pattern recognition was performed by using SPSS 25.0 and SIMCA-P 14.1 software（cluster analysis ，principal component analysis and orthogonal partial least squares regression analysis ）. The markers which affected the difference in the quality between P. asiatica and fried P. asiatica were screened with variable importance projection（VIP）value greater than 1. RESULTS There were 18 common peaks in the fingerprints of 15 batches of P. asiatica and 13 common peaks in the fingerprints of 15 batches of fried P. asiatica . A total of 8 common peaks were found in both of them. Their similarities were greater than 0.920. Two common peaks were identified as geniposidic acid ，acteoside. The results of cluster analysis showed that when the spacing was 10，the 30 batches of samples could be clustered into three categories ，with S 1-S5 as one，S16-S20 as one ，S6-S15 and S 21-S30 as one . The results of the pri ncipal component analysis showed that the cumulative variance contribution rate of the first two principal components was 82.575％ . The results of the orthogonal partial least squares regression analysis showed that the VIP values of the three common peaks were greater than 1，namely peak E（acteoside）， peak D （geniposidic acid ） and peak G. CONCLUSIONS Established fingerprints are stable ，simple sina.com and rapid. It can be used for the quality evaluation of P. asiatica and fried P. asiatica ，by combining with analysis of chemical pattern recognition. Acteoside ，geniposidic acid and the component represented by peak G may be the markers affecting the difference in quality of P. asiatica and fried P. asiatica .
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OBJECTIVE：To study the toxicity mechanism of yunacotine-induced arrhythmia in rats. METHODS ：Totally 32 rats were randomly divided by random number table method into normal control group ，yunacotine low-dose and high-dose groups （0.09，0.14 mg/kg），aconitine group （positive control ，0.88 mg/kg），with 8 rats in each group. Administration groups were given the corresponding drugs once a day ，and normal control group was given the constant volume of normal saline ，for consecutive 7 d. After last intragastric administration ，the changes of electrocardiogram （ECG） were observed. The contents of adenosine triphosphate（ATP）in myocardial tissue and Ca 2+ in myocardial cells ，the activities of Na +-K+-ATPase and Ca 2+-Mg2+-ATPase as well as the protein expression of ranolidine receptor 2（RyR2）and Ca 2+-ATPase（SERCA2）in myocardial tissue were determined. RESULTS：Compared with normal control group ，time limit of QRS wave and QTc intervals of rats were prolonged significantly in yunaconitine low-dose group （P＜0.01）. The content of Ca 2 + in myocardial cells ， the ATP contents ， the activities of Ca2+-Mg2+-ATPase and Na +-K+-ATPase as well as the protein expression of SERCA 2 in myocardial tissue were reduced significantly （P＜0.05 or P＜0.01）. The heart rate of rats in yunaconitine high-dose group and aconitine group were increased significantly （P＜ 0.05 or P＜0.01），and time limit of QRS wave and QTc intervals were significantly prolonged （P＜0.01）；the content of Ca 2+ in myocardial cells was increased significantly （P＜0.01）；ATP content ，the activities of Ca 2+-Mg2+-ATPase and Na +-K+-ATPase，and protein expression of RyR 2 and SERCA 2 in myocardial tissue were decreased significantly （P＜0.01）. CONCLUSIONS ： Yunaconitine can induce arrhythmia in rats ，the mechanism of which may be associated with Ca 2+ overload that resulted from reducing the activities of Na +-K+-ATPase and Ca 2+-Mg2+-ATPase and down-regulating the expression of related calcium transporter RyR2 and SERCA 2.
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OBJECTIVE：To study the effect of isofla vaspidicacid PB （called PB for short ）on the biofilm adhesion and the gene expression of ergosterol metabolism related enzymes in Trichophyton rubrum . METHODS ：M38-A2 method was adopted to determine MIC of PB to T. rubrum . MTT assay was used to screen the biolfilm condition and initial adhesion period of T. rubrum . The effects of different concentrations of PB （40，80，160 µg/mL）on the adhesion duration of T. rubrum （growth control group without PB was set up ，similarly hereinafter ）were evaluated and the adhesion rate was calculated by using XTT assay ；the effects of different concentrations of PB （20，40，80 µg/mL）on the biofilm formation of T. rubrum at different initial adhesion periods （3，5，9 h）were observed and the adhesion rate was calculated by using XTT assay combined with inverted microscope ；qRT-PCR method was used to detect the effects of PB （320 µg/mL）on the mRNA expression of ergosterol metabolism related enzyme gene ERG6 and ERG11 in biofilm of T. rubrum . RESULTS ：MIC of PB to T. rubrum was 20 µg/mL. The biofilm of T. rubrum in RPMI-1640 medium containing 10％ FBS was the most metabolism activity at 6 h of initial adhesion. Compared with growth control group ，after treated with different concentrations of PB ，adhesion rate and mRNA expression of ERG6 and ERG11 in biofilm were decreased significantly （P＜0.01）. Hyphae decreased or even disappeared ，and the adhesion inhibition rate （at 5 and 9 h of initial adhesion ）increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：PB can inhibit the adhesion of T. rubrum and reduce the hyphae ；the mechanism may be associated with the inhibition of the biofilm adhesion and mRNA expression of ergosterol metabolism related enzyme gene ERG6 and ERG11.
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OBJECTIVE：To study the effects of different penetration enhancers on in vitro transdermal permeation of Flavaspidic acid BB cream. METHODS ：Flavaspidic acid BB cream was prepared ，containing 11 kinds of different penetration enhancers as 1％ azone，2％ azone，3％ azone，4％ azone，1％ menthol，1％ propylene glycol ，1％ oleic acid ，1％ azone+1％ menthol，1％ azone+1％ propanediol，1％ azone+1％ oleic acid or 1％ menthol+1％ propanediol. Modified Franz diffusion cell was adopted using abdominal skin of isolated male rat as transdermal barrier. The content of flavaspidic acid BB was determined by UPLC. The accumulative transdermal amount （Q24 h）and percutaneous permeability （Jss）within 24 h were calculated ；and compared with Flavaspidic acid BB cream without transdermal enhancer ，the enhancement ratio （ER）was calculated. RESULTS ： Q24 h of Flavaspidic acid BB cream with above 11 kinds of transdermal enhancers were （82.96±7.15），（80.17±0.66），（78.22± 1.87），（73.53±1.24），（35.65±2.23），（34.02±1.73），（42.68±2.66），（33.94±1.37），（34.16±1.54），（46.78±1.21），（43.66±1.69） μg/cm2，respectively. Jss value were （5.26±0.10），（4.69±0.12），（4.45±0.45），（4.00±0.06），（3.74±0.33），（3.23±0.18）， （3.73±0.53），（3.14±0.47），（3.54±0.11），（3.98±0.34），（4.34±0.14）μg（/ cm2·h），respectively. ER were 2.055，1.831，1.738， 1.564，1.462，1.263，1.456，1.227，1.385，1.557，1.698，respectively. CONCLUSIONS ：All of the above transdermal absorption enhancers can enhance the percutaneous absorption of Flavaspidic acid BB cream ，among which ，1％ azone is the best.
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Objective: To observe the relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia and evaluate the safety preliminarily.Methods: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia were studied by the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The acute toxicity test and maximum tolerance test were carried out to evaluate the safety.Results: The total alkaloid in Atalantia buxifolia at low dose could obviously prolong cough incubation period and decrease cough times in mice, and that at high dose could significantly increase the secretion of phenol red in respiratory tract, and compared with those in the blank group, the differences were statistically significant (P0.05).Conclusion: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia are notable in the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The maximum tolerable dose test shows the total alkaloid in Atalantia buxifolia is safe.
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Objective:To evaluate the protective effect of Diclipterachinensis polysaccharide P2B on liver cell line L-02 injury in-duced by carbon tetrachloride ( CCl4 ) . Methods:The human liver L-02 cells were cultured, and the injury model was built by CCl4 . The L-02 cells were divided into the normal control group, the CCl4-damaged group, and the P2B sample groups (0. 125, 0. 250 and 0. 500 mg· ml-1 ). The contents of alanine aminotransferase ( ALT), aspartate aminotransferase ( AST), superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by MTT assay. Results:Compared with the CCl4-damaged group, P2B could improve the activity of L-02 cells, and the activity of AST and ALT in the supernatant was significantly reduced, and the content of SOD in the cells was increased and that of MDA was decreased. Conclusion:P2B can significantly prevent L-02 cells from the damage induced by CCl4 in a dose-dependent manner, and the mechanism may be related with the anti-oxidative activity of P2B.
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Objective:To isolate and prepare the reference substances of phloroglucinol derivatives from Dryopteris fragrans by semi-preparation HPLC. Methods:After reflux extraction of Dryopteris fragrans with petroleum ether,the extracting solution was con-centrated and separated by silica gel column chromatography,and then isolated by semi-preparation HPLC. The isocratic elution was carried out using acetonitrile as the mobile phase at 3. 0 ml·min-1 and the injection volume was 0. 5 ml. Two phloroglucinol deriva-tives were isolated. Results:The chemical structure of the two phloroglucinol derivatives respectively was aspidin BB with the purity of 98. 81% and aspidin PB with the purity of 98. 57% by ultra high performance liquid chromatography. Conclusion:The isolation of as-pidin BB and aspidin PB by semi-preparation HPLC is simple and fast with the purity over 98%,which can be used to prepare the ref-erence substances.
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Objective:To establish the quality standard for Yunüjian granules. Methods:Rehmannia glutinosa, rhizoma anemar-rhenae and achyranthes in the granules were identified by TLC. The HPLC method was adopted. A SunfireTM C18 column (250 mm × 4. 6 mm, 5 μm) was used with the mobile phase consisting of acetonitrile-0. 1% acetic acid solution(16∶84) at the column tempera-ture of 30 ℃. The flow rate was 1. 0 ml·min-1 ,the detection wavelength was 334nm, and the imjection volume was 10 μl. Results:The TLC spots of verbascoside, and sarsasapogenin in rhizoma anemarrhenae and cyasterone in achyranthes were quite clear with good seperation. The linear range of verbascoside was within the range of 0.163-0.612 μg(r=0.999 9), and the average recovery was 101. 3%(RSD=2. 7%,n=6). Conclusion:The method is simple, reliable and accurate, and can be applied in the quality control of Ynnüjian granules.
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Objective To explore the epidemic trend of influenza and pandemic virus variation in Maanshan, and to provide scientific basis for the prevention of influenza. Methods The ILI data of monitor hospitals and some ILI nasopharyngeal specimens were collected, then influenza virus was isolated by MDCK cells, and influenza serotypes were identified by HI. Results 15 904 cases of ILI were reported from 2006 to 2007. The detectable rate of influenza like illness in outpatients was 6.63%. 2 656 nasopharyngeal specimens of ILI were collected, and 358 strains of influenza virus were isolated, with isolation rate of 13.48%, of which 142 were H1N1 subset influenza A virus, 49 were H3N2, 72 were Victoria subset influenza B virus, and 96 were Yamagata. Conclusions During 2006 and 2007 two peaks of influenza-like illness in outpatients was in winter-spring and summer seasons of each year. More attention meeds to be paid to the monitoring of influenza.
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Objective To establish an HPLC method to determine simultaneously the content of six water-soluble chemical constituents (danshensu,protocatechuic aldehyde,caffeic acid,rosmarinci acid,salvianolic acid B,and salvianolic acid A) in Xiangdan Injection,and to compare their content discrepancy of Xiangdan Injection from 20 manufacturers.Methods The HPLC method was established.The column was Alltima C18 (250 mm?4.6 mm,5 ?m) at the gradient elution mode with the flow rate of 1.0 mL/min,column temperature was kept at 25 ℃ and the detector was set at 288 nm.Results Six components were separated clearly.The relationship between the concentration and the peak areas of the six compounds was all linear.The precision,stability,repetition,and average recovery were complied with the limit.Significant difference was found in contents of six chemical constituents in Xiangdan Injections from different manufacturers that the variation of salvianolic acid A content was the most distinct and danshensu was the most quietly.Conclusion The method has been successfully used to quanlify the six compounds in Xiangdan Injection,and then,can offer the reference for the quality control of Xiangdan Injections roundly.
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OBJECTIVE:To provide references and suggestions for the further development of pharmaceutical industry in Hangzhou city.METHODS:Using the classic normal form of SCP in the industry economics,i.e.market structure-the manufacturer behavior-market performance,a transverse and longitudinal comparative study was carried out on the market structure,the manufacturer behavior and the market performance of the Hangzhou's pharmaceutical industry between 2003 and 2006.RESULTS:Hangzhou's pharmaceutical industry was characterized by oligarch model in concentration degree,low differentiation in products;high barrier of entry and exit;low strength in advertisement,and rational in the merge and recombination.The manufacturers in this area attached increasingly importance to technological progress and the market performance was remarkable.CONCLUSIONS:Considering the status quo of Hangzhou's pharmaceutical industry,it is advisable for the manufacturers enlarge and strengthen their business,enhance its R&D investment and product differentiation,and stick to the market-oriented marketing concept.