OBJECTIVE:To establish a method for simultaneous determination of 4 effective components in the couple of Coptidis Rhizoma and Evodiae Fmctus,and to investigate rational proportion of the couple.METHODS:HPLC method was adopted.The determination was performed on Welch XB C18 column with mobile phase consisted of acetonitrile-1.5 mmol/L sodium dodecyl sulfate solution (pH adjusted to 5.0 with phosphoric acid) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 265 nm,and the column temperature was 30 ℃.The sample size was 10 μL.RESULTS:The linear ranges of berberine hydrochloride,martin hydrochloride,evodiamine and rutacarpine were 2.24-44.80 μg/mL(r=0.999 9),1.26-31.50 μg/mL(r=0.999 8),2.70-81.00 μg/mL(r=0.999 8),1.65-49.50 μg mL(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.The recoveries were 98.11％-100.73％ (RSD=1.04％,n=6),96.54％-103.47％(RSD=1.86％,n=6),95.49％-102.36％ (RSD=2.05％,n=6),97.19％-103.24％ (RSD=2.19％,n=6),respectively.When the proportion of Coptidis Rhizoma-Evodiae Fructus(m/m) was 2∶ 1,the comprehensive content of each component was the highest.CONCLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for simultaneous determination of 4 effective components in different compatibility proportions of the couple of Coptidis Rhizoma and Evodiae Fructus.The rational proportion of the couple of Coptidis Rhizoma and Evodiae Fructus is 2 ∶ 1.

Objective To evaluate the effect of LC+LCBDE and EST+LC in treating the cho-leeystolithiasis with choledocholithiasis. Methods The clinical data of 256 patients treated in our hos-pital were retrospectively analyzed. Of the 256 patients, 132 were treated by LC+LCBDE and 124 by EST combined with LC. The clinical data of the two groups was compared in operation success rate, operation time and cost, complication rate and operative hospital stay. Results There was no statisti-cal difference in the operation success rate, complication rate, operative hospital stay between 2 groups. However, there were significant differences in the operation time and cost between the 2 gruops. Conclusion There are respective indications, advantages and disadvantages in the two groups. EST+LC is the better choice for patients with a CBD<1.0 cm in diameter, stones impacted in the distal CBD, or old age. Otherwise, the better way is LC+LCBDE for patients with a CBD>1. 0 cm in diameter and multiple choledocholithiasis, especially in young and middle-aged people.

Objective To evaluate the effect of laparoscopic cholecystectomy(LC) combined with bile duct exploration and stone removal(BDE) and LC with endoscopic sphincterotomy(EST) in treating cholecystolithiasis with choledocholithiasis.Methods Among 256 cases of cholecystolithiasis and choledocholithlasis,132 patients were treated by LC+LCBDE,and 124 cases by EST combined with LC.The operation success rate,operation time and cost,complication rate,and length of hospital stay of the two groups were compared.Results There was no statistical difference in the operation success rate,complication rate,stone clearance rate,and average hospital stay between the two groups,but EST+LC group had significantly longer operation time and higher cost.Conclusions There are respective indications,advantages and disadvantages in the two groups.EST+LC is the better choice for patients with diameter of CBD1.0cm and with multiple choledocholithiasis,especially for middle-aged patients,the better way is LC+LCBDE.

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Objective To study the effect of Atorvastatin on the level of serum high-sensitivity C-reactive protein(hs-CRP) and nitric oxide(NO) in patients with acute cerebral infartion(ACI). Methods 60 patients with ACI were randomly divided into the Atorvastatin therapy group (n=30) and the conventional therapy group (n=30). At the basic of conventional therapy, the Atorvastatin therapy group was treated with Atorvastatin 20 mg/d. Both groups were treated for 14 consecutive days. The levels of serum hs-CRP and NO were measured and the scores of neurological deficit (NDS) were evaluated before and after treatment. Results The levels of serum hs-CRP in both two groups after 14 d treatment were significantly lower than those before treatment ( all P

Objective To observe the effects of low-dose theophylline and tiotropium on lung function and quality of life in patients with mild-moderate stable chronic obstructive pulmonary disease (COPD) and evaluate its safety.Methods This was a randomized,parallel-group,controlled trial.A total of 115 patients with mild-moderate stable COPD were divided into tiotropium group (37 cases),slow-release theophylline group (40 cases) and the combination of slow-release theophylline and tiotropium group (38 cases) by random digits table method.Thirty-eight patients without cardiopulmonary diseases were enrolled in control group.Observation period was 12 months.The lung function,6 min walking test distance (6MWD),modified British Medical Research Council Scale (mMRC),and COPD assessment test (CAT) were monitored before treatment and after treatment for 12 months.Results Of 115 patients,107 patients (35cases in tiotropium group,36 cases in slow-release theophylline group and 36 cases in combination of slowrelease theophylline and tiotropium group) completed the study.No significant difference was found in spirometry parameters reflecting airflow limitation after 12 months treatment compared with that before treatment in tiotropium group,slow-release theophylline group and the combination of slow-release theophylline and tiotropium group (P ＞ 0.05),such as the percentage of forced expiratory volume in 1 second (FEV1) over the expected value (FEV1％) and FEV1/forced vital capacity (FVC).The percentage of mid expiratory flow over the expected value (FEF25～75％) was improved in all groups,but the increment of FEF25～75 ％ was much higher in tiotropium group and the combination of slow-release theophylline and tiotropium group than that in slow-release theophylline group:(39.23 ± 7.77)％,(39.99 ± 8.25)％ vs.(34.91 ± 9.50)％,there were significant differences (P ＜ 0.05).Similar changes were observed in mMRC and CAT score.There was significant difference in CAT score between tiotropium group,the combination of slow-release theophylline and tiotropium group and slow-release theophylline group:(14.34 ± 2.22),(14.39 ± 3.53) scores vs.(16.22 ± 3.35) scores,P ＜ 0.05.6MWD was no obvious change in tiotropium group,slow-release theophylline group and the combination of slow-release theophylline and tiotropium group before and after treatment.The use frequency of short-acting drugs bronchiectasis was the lowest in the combination of slow-release theophylline and tiotropium group,and only was (2.3 ± 1.4) times per week.Fourteen patients happened COPD exacerbations in slow-release theophylline group during 12 months treatment.The duration in slow-release theophylline group was more than that in tiotropium group and the combination of slow-release theophylline and tiotropium group:(9.76 ± 2.25) d vs.(7.85 ± 2.51),(8.29 ± 2.24) d,and there was significant difference (P ＜ 0.05).Conclusions For mild-moderate stable COPD patients,the quality of life and dyspnea scores are not improved significantly after treatment of low-dose slow-release theophylline,and the combination slow-release theophylline and tiotropium may be more beneficial and safe.

Objective To investigate the impacts of c.388A ＞ G polymorphism of the solute carrier organic anion transporter 1B1 (SLCO1B1) gene on lipid-lowering and anti-atherosclerosis effects of atorvastatin in Chinese patients with ischemic stroke.Methods The patients with ischemic stroke whose baseline low-density lipoprotein cholesterol (LDL-C) ＞ 1.8 mmol/L were enrolled prospectively.They received atorvastatin (20 mg/d) for 12 months.The lipid and bilateral carotid intima-media thickness (CIMT) were measured respectively before and after treatment.The CIMT differences between SLCO1B1 c.388A＞G genotype groups were compared.Results A total of 71 patients with ischemic stroke were enrolled,including 5 AA genotype,31 AG genotype,and 35 GG genotype.The A allele frequency was 28.9％ and the G allele frequency was 71.1％.After treatment,the total cholesterol (TC),triglyceride (TG),and LDL-C in all patients were significantly lower than those before treatment,and high-density lipoprotein cholesterol (HDL-C) was significantly increase (all P＜0.001),but CIMT did not have significant change (P=0.475).The proportion of patients whose LDL-C ＜ 1.8 mmol/L or LDL-C decreased ≥50％ in the GG genotype group was significantly higher than the AG + AA genotypes group (74.29％ vs.44.44％;x2 =6.540,P =0.011).Conclusions SLCO1B1 gene c.388A ＞ G polymorphism could influence the lipidlowering effect of atorvastatin,lipid-lowering effect in the GG genotype group was better than that in the AG+ AA genotype group.SLCO1B1 gene c.388A ＞ G polymorphism did not have effect on the antiatherosclerosis effect of atorvastatin,but it might be associated with too short follow-up time.

Objective: To explore the impact of knocking out wildtype p53 phosphatase 1 gene on heart function with the changes of cardiac tissue mRNA and protein expressions in experimental mice. Methods: Our research included in 2 groups: Wildtype (WT) mice group and Wip1 knockout (Wip1-KO) mice group. n=10 in each group. The heart function, ratio of heart weight to body weight (HW/BW) were examined and compared between 2 groups; cardiac tissue morphology was observed by HE staining; mRNA expressions of ANP, BNP, MCP-1 andα-SMA were determined by RT-PCR and protein expressions of Bcl-2, Bax and c-caspase3 were measured by Western blot analysis. Results: Compared with WT mice group, Wip1-KO mice group showed decreased Wip1 mRNA expression,P<0.05, decreased LVEF, LV fraction shortening and increased left ventricular end systolic diameter (LVESD), allP<0.05; Wip1-KO mice group had reduced BW and elevated ratio of HW/BW, bothP<0.05 even the heart weight was similar between 2 groups. There was no difference in cardiac tissue morphology between 2 groups; mRNA expressions of ANP, BNP, MCP-1 and α-SMA were similar between 2 groups,P>0.05; apoptosis related protein expressions of Bax/Bcl-2 and c-caspase3 were similar between 2 groups,P>0.05. Conclusion: Wip1 gene knockout may impair the heart function in experimental mice, while the relevant mechanism should be further investigated.

Objective To investigate the effect of reptroperitoneal laparoscopic operation on the parameters of platelet, D-dimer and thrombomodulin(TM). Methods Forty cases were divided into two groups according to the operative way, retroperitoneal laparoscopic operation (n= 20) and open operation (n=20). Blood samples were taken preoperatively and at the end of the surgery. The following parameters were measured and compared within each group and between groups: platelet count (PLT), mean platelet volume (MPV), platelet distributionwidth(PDW), D-dimer, TM. ResultsThere were no significant differences for the PLT, PDW, MPV, TM and D-dimer between before and after operation in each group. There was no difference between 2 groups either for all these indicators.No patients from either group suffered thrombosis or abnormal bleeding as a pastoperative complication. Conclusion Compared with the conventional operation, retroperitoneal laparoscopic operation dioesd not induce more change on parameters of platelet, D-dimer and TM.

AIM: To discuss the effects of phytoestrogenic-genistein on cathepsin K (CK) expression stimulated by interleukin-1α (IL-1α) in osteoclast-like cells (OCLs) . METHODS: The OCLs were isolated from tissue of human giant cell tumor of bone (GCT) . The cells treated with reagents were divided into 7 groups including control (treated with phenol red-free-DMEM), vehicle (treated with 1.2 nmol· L-1 IL- 1α), 10-10-10-6genistein, genistein+ ICI 182.780, and 17[β-estrodiol (17β-E2) group. The cells were treated with 1.2 nmol· L-1IL-1α after pre-treated with genistein or 17β-E2 for 48 h (excluded the control group) . Expression of CK wasdetermined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot in OCLs stimulated by IL-1α in the presence of genistein or 17[β-E2. RESULTS: The obvious increase of expression of CK by IL1α in vehicle group was noted in comparing with control group (P ＜ 0.01 ) . Genistein down-regulated CK gene expression stimulated by IL-1α at the transcription level in a dose-dependent manner (r = 0.68, P ＜ 0.01 ) .Genistein down-regulated CK protein expression stimulated by IL-1α also in a dose-dependent manner (r = 0.61,P ＜ 0.01 ). The effects of genistein were abrogated partly after treatment with the estrogen receptor antagonist ICI 182.780. CONCLUSION: Genistein inhibits CK expression stimulated by IL-1α partly through estrogen receptor in OCLs.