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 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.

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.

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.

Objective:To analyze the clinical characteristics of neonatal enteroviral meningitis.Methods:A retrospective analysis was conducted on 87 cases with intracranical infection.The cerebrospinal fluid(CSF)samples of these neonates were collected and tested by polymerase chain reaction(PCR) for enterovirus.According to the results, the cases were divided into the positive group(55 cases) and negative group(32 cases). The clinical characteristics of the cases were collected.Results:The time in hospital of the positive group was shorter than the negative group[(9.5±5.2)d vs.(21.8±15.8)d, t=4.295, P<0.001]. The cure rates of the positive group was higher[96.4%(53/55) vs. 81.2%(26/32), χ2=3.872, P=0.049]. There was no death case in the positive group, while there were 3 death cases(9.4%, 3/32)in the negative group.In summer and autumn, the morbidity of the positive group was higher[96.4%(53/55) vs.62.5%(20/32), χ2=17.181, P<0.001]. In two groups, the most predominant clinical presentation was fever[90.9%(50/55) vs. 68.8%(22/32), χ2=6.962, P=0.008]. CSF examination results showed that white blood cells, the proportion of polymorphonuclear cell and CSF protein in the positive group were lower than those in the negative group, while the proportion of mononuclear cell and CSF glucose level in the positive group were higher( P<0.001). Blood and CSF cultures were negative in the positive group.There were 10 cases whose blood cultures and CSF cultures were positive in the negative group.Among them, there were 3 cases infected by group B streptococcus, 6 cases by E. coli, 1 case by Chryseobacterium meningosepticum.The prognoses of the CSF culture positive cases were poor.The abonormal cases detected by cerebral ultrasound, MRI and aEEG were fewer in the positive group( P<0.05). Conclusion:Enterovirus is the common pathogen of nonbacterial meningitis in summer and autumn in our region.The prognosis of enteroviral meningitis is better.Neonates with suspected clinical presentation, such as fever, should undergo PCR test to check CSF for enteroviru.

Objective To explore the effect of interleukin-6 (IL-6) and Interleukin-12(IL-12) on immune response to hepatitis B vaccination in infants of HBsAg-positive mothers.Methods A total of 91 neonates whose mothers were HBsAg-positive were included and followed up for 12 months.HBV DNA and HBV serological markers in the peripheral blood of the neonates and infants were detected with fluorescence quantitative polymerase chain reaction (FQ-PCR) and chemiluminescence immunoassay (CLIA),and the levels of IL-6 and IL-12 in the peripheral blood of the neonates and infants were detected with enzyme-linked immunosorbent assay (ELISA).Results The non-/hypo-response rate to hepatitis B vaccination was 35.16％ (32/91) in the 91 infants.In the neonatal period and infantile period,the level of IL-6 in non-/hypo-response group was lower than that in high-response group,while the level of IL-12 was higher than that in high-response group,and there was significant difference (P＜0.01).From the neonatal period to the infantile period,the level of IL-6 increased,while the level of IL-12 descended in both groups,and there was significant difference (P＜0.01).Furthermore,the level of anti-HBs of infants was positively correlated with the level of IL-6 (rs =0.70,0.79,P＜ 0.01),and was negatively correlated with the level of IL-12 (rs=-0.71,-0.72,P＜0.01) in the neonatal period and the infantile period.From the neonatal period to the infantile period,the increased level of IL-6 was positively associated with the level of anti-HBs (rs =-0.74,P＜0.01),while the decreased level of IL-12 was negatively associated with the level of anti-HBs (rs=-0.42,P＜0.01).The level of IL-6 was negatively correlated with the level of IL-12 in the neonatal period and the infantile period (rs=-0.68,-0.70,P＜0.01).Conclusions IL-6 might promote the immune response to hepatitis B vaccination in infants whose mothers were HBsAg-positive,while IL-12 might inhibit the immune response.IL-6 and IL-12 would affect the immune response to hepatitis B vaccination in infants of HBsAg-positive mothers at the same time.