OBJECTIVE: To develop a method for determination of atractylenolide Ⅰ and atractylone in Chang’ankang granule by HPLC. METHODS: The determination was performed on Yilite ODS column (250 mm?4.6 mm,5 ?m). The mobile phase contained of acetonitrile-water (gradient elution) with flow rate of 1 mL?min-1 and detection wavelength of 220 nm. RESULTS: The linear range of atractylenolide Ⅰwere 0.1～2.0 ?g(r=0.999 9) with mean recovery of 98.84%(RSD=0.76%,n=6).The linear range of atractylone were 0.5～10.0 ?g with mean recovery of 98.43%(RSD=1.14%,n=6). CONCLUSION: This method is simple, convenience and accurate for quality control of Chang’ankang granule.

Objective:To discuss the effect of Rhizoma Alismatis on blood fat in different health volunteers. Methods:The experiment wasfinished in NO.9 People’s Hospital of Chongqing,from January 2007 to December 2007. 26 subjects were all healthy people. The 26 sub-jects took Rhizoma Alismatis granula(national food health product No.:G20040383,provided by the biochemistry technology development center of Shanghai). The essential component was Rhizoma Alismatis,5 g/pouch,taken once every morning and night,one pouch once time for two weeks as a course of treatment,two courses. Blood was drawn from the volunteers under the age of 50 years and above the age of 50 years before and after medication for two and four weeks,respectively. Serum protein cholesterol,apoprotein A,apoprotein B were detected by Hitachi 7 600 automatic biochemistry analyzer. Results:In the volunteers under the age of 50 years,the level of triacylglycerol had a down tendency after 2 week medication. Compared with that before medication,the level of total cholesterol and low density lipoprotein cholesterol had significant difference (P

OBJECTIVE:To compare the distribution of levofloxacin in lung of mice after intragastric and intravenous adminis-tration of Tanreqing injection combined with levofloxacin. METHODS:144 mice were randomly divided into experimental group and control group,with 72 mice in each group. Both groups were given Tanreqing injection(0.8 ml/kg). 1 h later,control group was given Levofloxacin injection(80 mg/kg)intravenously,and experimental group Levofloxacin tablet solution(80 mg/kg)intra-gastrically;every 8 mice were sacrificed for lung tissue collection at the time points of 0.05,0.083,0.17,0.5,1.0,2.0,4.0,8.0, 12.0 h and 0.083,0.17,0.33,0.5,1.0,2.0,4.0,8.0,12.0 h,respectively. Using ciprofloxacin as internal standard,the content of levofloxacin in lung tissue was determined by HPLC,and pharmacokinetic parameters were calculated by 3p97 program. RE-SULTS:The concentration-time curves of levofloxacin in lung tissue were in line with two-compartment model in 2 groups. The main pharmacokinetics of experimental group and control group were as follows as t1/2β of(4.17±0.84)h and(4.10±0.55)h;CL of(0.66±0.049)L/h and(0.71±0.21)L/h;AUC0-t of(109.48±12.34)mg·h/kg and(113.04±29.43)mg·h/kg;cmax of(38.76± 1.62) mg/kg and (42.28 ± 2.03) mg/kg,respectively. There was no statistical significance (P>0.05). Absolute bioavailability of levofloxacin was(96.85±17.39)% in experimental group with intragastric administration. CONCLUSIONS:After intravenous injec-tion of Tanreqing injection,the distribution of levofloxacin in lung tissue of mice are similiar between intragastric administration and intragastric administration.

OBJECTIVE:To optimize the formulation of entecavir PLGA sustained-release microspheres,and explore its drug release in vitro. METHODS:PLGA sustained-release microspheres was prepared by emulsification-solvent evaporation method. Us-ing composite score of entrapment efficacy and drug loading as indexes,orthogonal test was designed to optimize drug amount, drug-PLGA mass ratio,PLGA mass concentration,oil phase-aqueous phase volume ratio and polyvinyl alcohol (PVA) concentra-tion;and validation test was also conducted. The prepared microsphere morphology,particle size and durg release in vitro were de-tected. RESULTS:The optimized formulation was entecavir 20 mg,entecavir-PLGA mass ratio 1∶10,PLGA mass concentration 200 mg/ml,oil phase-aqueous phase volume ratio 1∶10,and PVA concentration 2%;entrapment efficacy was (86.52 ± 3.25)%, drug loading was(18.36±1.37)%,RSDs were lower than 5.0%(n=3);it was round and smooth in appearance with average par-ticle size of 58.35 μm;Q10 h,Q96 h and Q360 h were 9.6%,42.9% and 89.6%,and the drug release in vitro fitted to Higuchi model (r2=0.965 8). CONCLUSIONS:Entecavir PLGA sustained-release microspheres prepared by optimized formulation has good sus-tained-release performance.

Objective To investigate the feasibility of fine needle aspiration cytopathology in the diagosis of lymphoma.Methods To compare cytopathology with histopathology of 72 lymphoma cases and analyse immunochemistry stain of 3 cases.Results 61 of 72 cases were diagnosised as lymphoma in cytopathology,while 58 of 61 cases were diagnosised as lymphoma in histopathology.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value rates of cytopathology were 89.2 ％ (58/65),57.1％ (4/7),86.1％ (62/72),95.1％ (58/61),and 36.4 ％ (4/11) respectively.Conclusion Fine needle aspiration cytopathology is significant to the diagnosis of lymphoma.It should greatly improve the accuracy of lymphoma diagnosis and make it possible to subclassify lymphoma,combined cytopathology with other auxilary detection.

Objective To explore the difference of serum oxytocin levels between autistic children and healthy children at the age of 3-5,and the relationship between serum oxytocin levels and social competence in children with autism.Methods Twenty-five autistic children and twenty healthy children were tested by an enzyme-linked immunosorbent assay to evaluate the oxytocin levels;the SRS was used to evaluate social competence of children with autism.Results Mann-Whitney test showed there were significantly differences in oxytocin level between autistic children and healthy children(P<0.05).The social competence of autistic children was negatively correlated to oxytocin levels(r=-0.735,P<0.01).Multiple liner regression analyses showed that oxytocin level was an impact factor of the social competence of autistic children(F=11.931,P<0.01).Conclusion This study indicates that the serum oxytocin level maybe a factor which influence the social competence of autistic children.

Objective To analyze the epidemud growth factor receptor(EGFR)mutations in NSCLC patients in Fujian province.Methods Fresh specimens of lung cancer and corresponding normal lung tissue were collected from 50 cases of NSCLC patients.After DNA extraction,nested polymerase chain reaction (nested PCR)and direct deoxyribonucleic acid(DNA)sequencing were used to analyze EGFR gene mutations in NSCLC patients.Results EGFR mutations in tumors were identified from 13 of 50(26%)patients,including 10 cages of in-frame deletion in exon 19 and 3 cases of amino acid substitution in exon 21.Conclusion The mjor type of EGFR mutation in NSCLC patients in Fujian is in-frame deletion in exon 19.

Objective To investigate the molecular mechanism how edaravone reduces cytotoxicity caused by hypoxia/reoxygenation-induced injury.Methods Ischemia model-hypoxia/ reoxygenation model which also called oxygen and glucose deprivation/reoxygenation model (OGD-R) was established.Cells were divided into control group,OGD-R group,OGD-R with edaravone of different concentration groups Cells of OGD-R model were pretreated with the appropriate concentration of edaravone or combine with the specific channel blockers LY294002 and MK2206 individually,the expression levels of ERK,AKT and Bcl-2 were detected with Western-blot,while the expression levels of BDNF and Bcl-2 mRNA detected with RT-PCR,the expression levels of BDNF protein detected with ELISIA,the levels of LDH releasing and Hoechst33258 staining used with homologue assay kit or regents.Results The pretreatment of cultured neurons with edaravone alleviated hypoxia/reoxygenation induced neuronal injury in a dose-dependent manner (LDH releasing reduced,Caspase-3 activity and ratio of nuclear pyknosis declined).The edaravone pretreatment did not increase significantly the p-ERK expression levels,but the amount of p AKT expression was significantly increased(P＜0.01).Pretreatment of edaravone(10 μmol/L) remarkably reduced the LDH release and declined the ratio of nuclear pyknosis after reoxygenation (P＜0.01),which was inhibited by MK2206.Compared to OGD-R,edaravone pretreatment markedly promoted BDNF and Bcl-2 mRNA expression at each time point,and the most pronounced effects occured at 8 h after OGDR.ELISIA analysis showed that BDNF protein level was significantly elevated after edaravone pretreatment at the same point.Western blot analysis indicted that edaravone pretreatment significantly enhanced Bcl-2 protein expression at 8 h after OGD-R,which was blocked by MK2206.Conclusions Edaravone may alleviate hypoxia/reoxygenation induced neuronal injury by enhancing BDNF and Bcl-2 expression and inhibiting Caspase-3 activity through activation of the PI3K/Akt pathway.

Objective To investigate the advantages of detection for EGFR gene mutations by denaturing high performance liquid chromatography (DHPLC) technology. Methods DHPLC was used to detect EGFR gene mutations at exon 19 and 21 in 49 cases of non-small cell lung cancer (NSCLC) patients,and the direct DNA sequencing was used to verify the accuracy of DHPLC detection. Results EGFR gene mutation was identified from 13 of 49 cases by DHPLC,including deletion mutation at exon 19 in 10 cases (76.92 %) and alternative mutations at exon 21 in 3 cases (23.08 %). Mutation results of DHPLC was consistent with DNA direct sequencing. The results of the direct DNA sequencing were the same as those of DHPLC. The sensitivity of mutation test by DHPLC was 100 %. Conclusion DHPLC technology can be used for large scale screening of EGFR gene mutation with rapid and accuracy.

OBJECTIVETo investigate the methylation status of Runx3 promoter and Runx3 expression in breast lesion tissues.

METHODSOne hundred and fourteen breast lesions, including 35 cases of fibroadenoma, 39 cases of intraductal carcinoma, 40 cases of invasive ductal carcinoma, and 33 cases of normal breast tissue from Fabruary 2010 to August 2012 were included in this study. Runx3 protein expression was assessed by immunohistochemical SP method; whereas methylation of Runx3 promoter was assessed by high resolution melting (HRM) analysis.

RESULTSRunx3 protein was mainly expressed in the cytoplasm of ductal epithelial cells. The expression rates of Runx3 in normal breast tissue, fibroadenoma, ductal carcinoma in situ, invasive ductal carcinoma were 87.9% (29/33), 85.7% (30/35), 53.8% (21/39), and 40.0% (16/40) respectively. The methylation rates of Runx3 promoter were 12.1% (4/33), 20.0% (7/35), 46.2% (18/39), and 57.5% (23/40), respectively. Correlation analysis between promoter methylation and protein expression of Runx3 in different breast tissue showed the r value in normal breast tissue, fibroadenoma, ductal carcinoma in situ and invasive ductal carcinoma was -0.431 (P = 0.012), -0.408 (P = 0.015), -0.589 (P = 0.000) and -0.743 (P = 0.000) respectively.

CONCLUSIONSRunx3 protein expression shows a downward trend in ductal carcinoma in situ and invasive ductal carcinoma, meanwhile its promoter methylation increases significantly. The methylation of Runx3 promoter may be one of the important factors in the occurrence and development of breast cancer.

Breast ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Fibroadenoma ; metabolism ; Humans ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic