OBJECTIVE:To establish HPLC method for the content determination of oleanolic acid and ursolic acid in Prunella vulgris with ?-CD as additive agent of mobile phase.METHODS:The sample was Prunella vulgaris.The separation was performed on a Tigerkin ODS3(250 mm?4.6 mm,5 ?m) column with mobile phase consisted of methanol-10 mmol?L-1 ?-CD 1% ammonium acetate(86:14) at flow rate of 1.0 mL?min-1.The column temperature was set at 40 ℃ and UV detection wavelength was 210 nm.RESULTS:The linear range of oleanolic acid and ursolic acid were 0.5～5.0 ?g(r=0.999 2) and 1.0～10.0 ?g(r=0.999 8) respectively.High,medium and low average recovery of oleanolic acid were 97.3%(RSD=1.04%),98.0%(RSD=0.87%),101.6%(RSD=1.65%),respectively.Those of ursolic acid were 97.8%(RSD=1.17%),98.7%(RSD=0.93%),100.4%(RSD=1.72%),respectively.CONCLUSION:The new method is simple,accurate and reproducible for the quality control of P.vulgaris and simultaneous determination of oleanolic acid and ursolic acid.

OBJECTIVE:To estabish a RP-HPLC method for the content determination of forsythin of Yinqiao jiedu pills.METHODS:The determination was performed using SHIM-PACK VP-ODS column(150 mm ? 4.6 mm,5 ?m) with column temperature at 30 ℃.The mobile phase was acetonitril-water(25∶75) with flow rate at 1.0 mL?min-1.The detective wavelength was set at 277 nm.RESULTS:The linear range of forsythin was 0.453～3.481 ?g(r=0.999 8).The mean recovery was 99.50%(RSD=1.15%,n=6).CONCLUSION:The method was simple,rapid,accurate and suitable for the quality control of Yinqiao jiedu pills.

Objective To evaluate the quality of immunological diagnosis kits of schistosomiasis. Methods The national reference IgG serum of schistosomiasis was applied for evaluating eight immunological diagnosis kits of schistosomiasis. Results All eight diagnosis kits had no National Production Certificate issued by State Food and Drug Administration. Six kits had good quality with high precision. After the physical examination, two kits did not meet the quality standards. Conclusions The good and bad quality of immunological diagnosis kits of schistosomiasis used in market is intermingled. The quality of these kits should be monitored.

Objective To investigate the therapeutic effects and mechanism of Artemisia argyi ferment substance on systemic Candida albicans infection. Methods The model of systemic Candida albicans infection was established in immunosuppressed mice. The model mice were randomly divided into the model control,Artemisia argyi ferment substance( AAFS) at different doses(100,200,and 400 mg·kg-1 )and fluconazole group(20 mg·kg-1 ),30 mice in each. Mice in each treatment group were given therapeutic drugs by gavage for 5 consecutive days,twice daily. The survival of mice was determined 21 days after the model was set up. The serum levels of IFN-γand IL-2 were determined by ELISA. The proliferation activity of T lymphocyte in the spleen was detected by MTT assay. The number of living fungi in liver and kidney tissues was counted. Results Compared with the model control,AAFS at middle and high doses and fluconazole significantly increased the survival rate of mice,the serum levels of IFN-γand IL-2,and the proliferation activity of T lymphocyte in the spleen,but decreased the number of living fungi in tissues(P〈0. 01). Compared with low dose AAFS,middle and high doses of AAFS and fluconazole showed significantly different effect on each index(P〈0. 05 or P〈0. 01),but there was no difference among these groups(P〉0. 05). Conclusion AAFS at 200-400 mg·kg-1 has inhibitory effects on systemic Candida albicans infection in mice,the mechanism of which is related to increasing the proliferation of T lymphocyte in spleen and the levels of IFN-γand IL-2 in serum.

A rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometric ( RRLC-QTOF/MS) method was used to profile the metabolites of urine samples from atherosclerosis ( AS) patients and healthy controls and find the differential metabolites which could provide the scientific evidence to explain the pathogenesis and early disease diagnose. In the study, 15 AS patients ( age46. 84±2. 41 years) and 15 healthy controls ( age45 . 72±1 . 93 years ) was screened out by VaSera VS-1000 . The urine samples were analyzed by RRLC-QTOF/MS and the resulting data matrices were analyzed by multivariate statistical analysis ( Principal Component Analysis, PCA ) to find the potential biomarkers. The results showed that the urine samples of AS patients were successfully distinguished from those of healthy controls. Besides, a total of two significantly changed metabolites, uric acid and Guanidineacetic acid, had been found and identified as potential biomarkers, which suggested that the disorder of purine metabolism and amino acid metabolism played an important role in the mechanism of AS.

OBJECTIVE:To provide reference for the establishment and improvement of work system for clinical pharmacist in PIVAS of our province even the country. METHODS:Questionnaire was conducted to investigate the PIVAS directors in the 65 third-grade class A hospitals in China,and the data was statistically analyzed. RESULTS:Totally 65 questionnaires were sent out, 63 were effectively received,with effective rate of 96.9%. 98.4% respondents thought PIVAS should equip clinical pharmacist,but the actual situation in respondent's hospital was not ideal. 100% thought clinical pharmacist in PIVAS should have the ability of prescription checking and prescription evaluation;more than 95% thought clinical pharmacist in PIVAS should assume the medi-cine publicity and education and need to have good communication with clinic;only 42.9% thought clinical pharmacist in PIVAS need to take part in the routine drug dispensing. CONCLUSIONS:It is necessary to equip clinical pharmacist with professional training in PIVAS. Clinical pharmacist in PIVAS should focus their ability on prescription checking and evaluation,medicine public-ity and education,communication with clinic.

Aim To discuss the effectsof extract from fermented buckwheat flower and leaf(EFBFL) on myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice and its mechanism.Methods 9-week-old male db/db mice were randomly divided into high level EFBFL dose group(EFBFL-H, 0.1 g·kg-1), low level EFBFL dose group(EFBFL-L, 0.05 g·kg-1),metformin hydrochloridecontrol group, model control group, and normal control group, with 10 mice in each group.All groups were treated with 8 wks of drugs by gastric perfusion.The random blood glucose(RBG) was tested respectively at the end of 2nd, 4th, 6th, and 8th wk.Finally, the levels of creatine kinase(CK) creatine kinase MB(CK-MB), andadvanced glycosylation endproducts(AGEs) were detected after 8 wks.The morphological changes of myocardium were observed under light microscope by HE staining, and the ultrastructure of myocardium was observed under electron microscope.Immunohistochemical method and Western blot were used to detect myocardial tissue glucose transporter-4(Glut-4).Results EFBFLcould repress patho-proceeding of myocardial fibrosis efficiently, and significantly decrease the level of blood glucose, CK,CK-MB, and AGEs in db/db mice.Meanwhile, it could increase the expression of Glut-4 in myocardial tissues of mice.Conclusions EFBFL can prevent myocardial injury in spontaneously obese type Ⅱ diabetic db/db mice.The possible mechanism may be related to lowering the level of blood glucose and serum AGEs and up-regulating Glut4of cardiac muscle.

Objective: To investigate the inlfuence of angiotensin converting enzyme 2 (ACE2) on lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) protein expression and to explore the protective effect of ACE2 on vascular endothelial cells. Methods: Our work includedin vitro andin vivo studies. For in vitro experiment, the human umbilical vein endothelial cell (HUVEC) were cultured and transfected with replication deficient recombinant adenovirus of ACE2 (Ad-ACE2), and LOX-1 protein expression stimulated by angiotensin 2 was examined by Western blot analysis. Forin vivo study, atherosclerosis plaques were induced in 20 apolipoprotein E-deifcient (ApoE-/-) mice, and then randomly divided them into 2 groups: ACE2 group, the mice received Ad-ACE2 (2.5×109 pfu/ml) injection through caudal vein, EGFP (enhanced green lfuorescent protein) group, the mice received equal replication deifcient recombinant adenovirus of EGFP (Ad-EGFP) injection through caudal vein.n=10 in each group. The animals were executed after 1 month treatment to collect abdominal aorta. Lipid content in atherosclerosis plaque was evaluated by Oil red O staining and LOX-1 protein expression was examined by immunohistochemistry and Western blot analysis. Results: Bothin vitro andin vivo experiments conifrmed that endothelial cell LOX-1 protein expression was signiifcantly inhibited by ACE2 transfection. The lipid content in ACE2 group was obviously lower than that in EGFP group byin vivo study. Conclusion: ACE2 may inhibit LOX-1 protein expression and therefore reduce the progress of atherosclerosis.

This study determined the effect of hepatitis B virus (HBV) replication in peripheral blood mononuclear cell (PBMC) from HBsAg-positive mothers on HBV intrauterine transmission.A total of 150 HBsAg-positive mothers and their neonates were recruited in this study.Within 24 h after birth,HBV serological markers,serum HBV DNA,PBMC HBV relaxed circular DNA (rcDNA),and covalently closed circular DNA (cccDNA) were measured in the HBsAg-positive mothers and their neonates before passive-active immune prophylaxis.The relationship between HBV replication in PBMC and HBV intrauterine transmission was examined through Chisquare test and logistic regression.The rate of HBV intrauterine transmission was 8.00％ (12/150) in the 150 neonates born to HBsAg-positive mothers.The positivities of PBMC HBV rcDNA and cccDNA in the HBsAg-positive mothers were 36.67％ (55/150) and 10％ (15/150),respectively.Maternal PBMC HBV cccDNA was a risk factor of HBV intrauterine transmission (OR =6.003,95％ CI:1.249-28.855).Maternal serum HBeAg was a risk factor of PBMC HBV rcDNA (OR =3.896,95％ CI:1.929-7.876) and PBMC HBV cccDNA (OR =3.74,95％ CI:1.186-11.793) in the HBsAg-positive mothers.Administration of hepatitis B immune globulin was a protective factor of PBMC HBV cccDNA (OR =0.312,95％ CI:0.102-0.954) during pregnancy.The positivity of PBMC HBV rcDNA was related to that of cccDNA in the HBsAg-positive mothers (x2=5.087,P =0.024).This study suggests that PBMC is a reservoir of HBV and an extrahepatic site for virus replication and plays a critical role in HBV intrauterine transmission.

Objective To construct a nonparametric proportional hazards (PH) model for mixed informative interval-censored failure time data for predicting the risks in heart transplantation surgeries. Methods Based on the complexity of mixed informative interval-censored failure time data, we considered the interdependent relationship between failure time process and observation time process, constructed a nonparametric proportional hazards (PH) model to describe the nonlinear relationship between the risk factors and heart transplant surgery risks and proposed a two-step sieve estimation maximum likelihood algorithm. An estimation equation was established to estimate frailty variables using the observation process model. I-spline and B-spline were used to approximate the unknown baseline hazard function and nonparametric function, respectively, to obtain the working likelihood function in the sieve space. The partial derivative of the model parameters was used to obtain the scoring equation. The maximum likelihood estimation of the parameters was obtained by solving the scoring equation, and a function curve of the impact of risk factors on the risk of heart transplantation surgery was drawn. Results Simulation experiment suggested that the estimated values obtained by the proposed method were consistent and asymptotically effective under various settings with good fitting effects. Analysis of heart transplant surgery data showed that the donor's age had a positive linear relationship with the surgical risk. The impact of the recipient's age at disease onset increased at first and then stabilized, but increased against at an older age. The donor-recipient age difference had a positive linear relationship with the surgical risk of heart transplantation. Conclusion The nonparametric PH model established in this study can be used for predicting the risks in heart transplantation surgery and exploring the functional relationship between the surgery risks and the risk factors.