0.05). Multivariate analysis revealed that risk factors for vascular complications included the difference in radial artery diameter and sheath size; prolonged procedure time and comorbidity of diabetes mellitus. Conclusion Transradial access is a safe and practical alternative approach for intervention procedure. Vascular ultrasonic examination is valuable for selection sheath before invention procedure especially in patients with diabetes mellitus and in assessment vascular complications after the procedure.

In this paper,based on the present situation of the pharmaceutical education in China,the author brings forward the challenge and opportunity to pharmaceutical education at medical colleges,and then gives us some helpful ideas to solve the problems.

AIM:To establish a method for determining saikosaponin A,paeoniflorin,hesperidin and ferulic acid in Chaihu Shugan Powder(Pericarpium Citri Reticulatae,Radix Bupleuri,Rhizoma Chuanxiong,Fructus Aurantii,Radix Paeoniae alba,etc.).METHODS:The chromatographic saparation was performed on a Hypersil C_(18) column(250 mm×4.6 mm,5 μm).The mobile phase was acetonitril-water(43:57)for saikosaponin A,and the mobile phase for rest components was acetonitrile-0.5% acetic acid(40:60).All of flow rates were 0.8 mL/min and column temperature maintained at 30℃.The detection wavelength was set at 200-400 nm.RESULTS:The four constituents were separated within 15 min.The linear ranges of saikosaponin A,paeoniflorin,hesperidin and ferulic acid were 38.5-166.7 μg/mL(r = 0.999 6),15.9～254.5 μg/mL(r = 0.999 9),22.1-353 μg/mL(r =0.999 3),6.30-201.5 μg/mL(r =0.999 9),respectively.The average recoveries were 97.57%,97.40%,98.86%,96.37%,respectively.The RSD were 2.1%,1.1%,0.70%,1.3%,respectively.CONCLUSION:The method is rapid,simple and accurate,and can be used for quality control of Chaihu Shugan Powder.

AIM:To establish a method for determining saikosaponin A,paeoniflorin,hesperidin and ferulic acid in Chaihu Shugan Powder(Pericarpium Citri Reticulatae,Radix Bupleuri,Rhizoma Chuanxiong,Fructus Aurantii,Radix Paeoniae alba,etc.).METHODS:The chromatographic saparation was performed on a Hypersil C18 column(250 mm?4.6 mm,5 ?m).The mobile phase was acetonitril-water(43:57) for saikosaponin A,and the mobile phase for rest components was acetonitrile -0.5% acetic acid(40:60).All of flow rates were 0.8 mL/min and column temperature maintained at 30 ℃.The detection wavelength was set at 200-400 nm.RESULTS:The four constituents were separated within 15 min.The linear ranges of saikosaponin A,paeoniflorin,hesperidin and ferulic acid were 38.5-166.7 ?g/mL(r=0.999 6),15.9～254.5 ?g/mL(r=0.999 9),22.1-353 ?g/mL(r =0.999 3),6.30-201.5 ?g/mL(r=0.999 9),respectively.The average recoveries were 97.57%,97.40%,98.86%,96.37%,respectively.The RSD were 2.1%,1.1%,0.70%,1.3%,respectively.CONCLUSION:The method is rapid,simple and accurate,and can be used for quality control of Chaihu Shugan Powder.

BACKGROUND: Taohong Siwu Tang is made up of Taoren, Honghua,Danggui, Chishao and Chuangxiong. Paeoniflorin is one of the major effective components in Radix Paeoniae Rubra Extracts and Taohong Siwu Tang, and it can promote blood circulation and remove blood stasis and has the effect of anti-acute myocardial ischemia.OBJECTIVE: To compare the serum concentration of paeoniflorin in mice after intragastric administration (i.g.) of Radix Paeoniae Rubra extracts and Taohong Siwu Tang.DESIGE: Rndomized controlled observation.SETTING: Department of Pharmacy and Department of Basic Medicine,Hebei North University.MATERIALS: The trial was performed in the first laboratory of Department of Pharmacy, Hebei North University during February to June 2005.Eighty involved Kunming mice of either gender and of clean grade, with body mass of 18 to 22 g, were purchased from Experimental Animal Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences. Taoren, Honghua, Danggui, Chishao, Shengdi and Chuanxiong 6 Chinese herbs were all purchased from Zhangjiakou Institute of Traditional Chinese Medicine.METHODS: ①Blank serum was isolated from eight mice which were fasted for 12 hours. Control sample solution of paeoniflorin was added into blank serum. Acetonitrile was used to deposit protein, the solution was centrifugated and isolated ,and 20μL supernatant was taken to obtain regression equation. ② Seventy-two Kunming mice were randomized into Radix Paeoniae Rubra extracts group and Taohong Siwu Tang group, with 36 mice in each. After being fasted for 12 hours, the mice in each group were intragastric administration of Radix Paeoniae Rubra extracts and Taohong Siwu Tang, respectively and the concentration of paeoniflorin was 300 mg/kg.1 mL blood was taken respectively at 15, 30, 60, 90, 120 and 180 minutes after intragastric administration and prepared into serum sample (6 mice at each time point in each group). The time to peak and difference in peak concentration of serum paeoniflorin were compared between intragastric administration of Radix Paeoniae Rubra extracts and Taohong Siwu Tang.The concentration of paeoniflorin was measured with high-performance liquid chromatography (HPLC). Chromatographic condition: chromatographic column YWG-C18 (250×4.6 mm,10 μm). The mobile phase was methanolKH2PO4 (0.05 mol/L) (38∶62) at a flow rate of 1.0 mL/min. The detective wavelength was set at 230 nm. The column temperature was 25 ℃.MAIN OUTCOME MEASURES: Serum concentration of paeoniflorin after administration of Radix Paeoniae Rubra extracts and Taohong Siwu Tang in mice.RESULTS: Seventy-two mice were used for Radix Paeoniae Rubra extr acts and Taohong Siwu Tang tests, and all of them entered the stage of result analysis without deletion. ① Chromatographic isolation results: Paeoniflorin could be well isolated from other components. The linear range for detection of paeoniflorin was 5.40-646.0 μg/mL. The mean recovery rate was 93.6% and the limit of detection was determined as 1.08 mg/L. ②Comparison of concentration of serum paeoniflorin of mice between two groups: The concentration of serum paeoniflorin in the Radix Paeoniae Rubra extracts group and Taohong Siwu Tang group reached the peak at 30 and 60 minutes after intragastric administration, respectively. The peak concentration of paeoniflorin of Taohong Siwu Tang group was lower than that of Radix Paeoniae Rubraextracts group [(36.27±5.72) mg/L vs.(46.82±5.29) mg/L, P ＜ 0.01 )].CONCLUSION: The concentration of serum paeoniflorin is significantly decreased after intragastric administration. It was further confirmed the decreased concentration of the crude paeonifiorin in the mice sera might result from promotion of its metabolic conversion by other chemical components in the complex prescription. This result needs further investigation.

ObjectiveTo study the change of autophagy of human pancreatic cancer cell MiaPaCa-2 before and after Ad5-KAI1 tranfection,and to investigate the possible mechanism.MethodsThe MiaPaCa-2 cells without KAI1 expression were infected with Ad5-KAI1 with KAI1 target gene,and Ad5-null was used as negative control,and parental cell was used as blank control.The formation of autophagosomes was observed by electromicroscopy.The green fluorescent protein-labeled light chain 3 (LC3) associations with autophagosome membranes was detected by confocal microscopy.PD98059,LY294002 were applied to pre-treat the cells.The expression levels of beclin 1,AKT,ERK,the phosphorylation of AKT and ERK protein and the ratio of LC3-Ⅱ to LC3- Ⅰ were detected by Western blotting.ResultsAfter 100 MOI Ad5-KAI1 infections for 24 h,the rate of cell expressing KAI1 protein reached (84.97 ±8.56)％,number of LC3 increased from 4 to 20; and swelling,degeneration of mitochondria was observed,and bilayer-like structure in cytoplasm was found.The expression of beclinl increased (1.4 ±0.3 ) folds,and the expression of LC3-Ⅱ/LC3- Ⅰ increased (8.00 ±2.78) folds.PI3K blockade LY294002 pretreatment significantly suppressed the phosphorylation of AKT of MiaPaCa-2 (2.756 vs 1.516),but it did not inhibit the increase of ratio of LC3-Ⅱ to LC3- Ⅰ(0.770 vs 1.403).ERK blockade PD98059 pretreatment not only significantly suppressed the phosphorylation of ERK of MiaPaCa-2 ( 1.637 vs 0.403 ),but also inhibit the up-regulation of beclin 1 protein expression ( 2.377 vs 1.150) and increase of ratio of LC3- Ⅱ to LC3- Ⅰ (2.225 vs 0.680).ConclusionsKAI1 can significantly induce autophagy of human pancreatic cell line MiaPaCa-2 through phosphorylation of ERK rather than AKT.

Objective To investigate the expression and significance of Smad4 and Smad7 in newborn rats with hyperoxia-induced chronic lung disease(CLD).Methods Sixty-four newborn Wistar rats 12 h after birth were divided into high-oxygen group (n =32) and air group (n =32,control group) by random number table method.The high-oxygen group was placed in the oxygen glass tank with continuous infusion of oxygen.And 1,3,7,14 d after experiment,tracheal separated,the chest opened to expose heart and lung,slices were Masson staining,undergo dynamic observation of the pulmonary pathological changes under light microscope.Lung fibrosis score was carried out to determine the degree of pulmonary fibrosis,and immunohistochemical technique was used to detect Smad4 and Smad7 protein expression in lung tissue.The expression levels of Smad4 and Smad7 protein in lung tissue were detected with Western blot.Results Compared with the air group,there was statistically significant difference in pulmonary fibrosis score on day 7 (2.67 ± 0.21 vs 0.58 ± 0.17) and day 14 (4.48 ± 0.24 vs 0.63 ± 0.13) in high-oxygen group (P ＜ 0.05) ; Smad4 and Smad7 was main in visible lung epithelial cells and interstitial fibroblasts.Smad4 expression in the high-oxygen group gradually enhanced,compared with the air group (P ＜ 0.05) on day 7 (122.35 ± 10.3 vs 140.08 ±7.77) and day 14(129.7 ± 7.33 vs 144.99 ± 6.49).Smad7 expression in the high-oxygen group first increased and then decreased,expression in the high-oxygen group increased on day 7 (122.35 ± 10.29 vs 130.56 ±9.8),and compared decreased with the air group(P ＜0.05) on day 14(132.16 ±4.38 vs 126.22 ±6.49).Conclusion The newborn rat exposed hyperoxia,the up-regulation of Smad4 protein expression and the down-regulation of Smad7 protein expression are imposible closely related to the happen and development of CLD pulmonary fibrosis.

Objective To explore the pathway of KAI1 induced autophagy regulating apoptosis in human pancreatic cancer cell line MiaPaCa-2.Methods There were three groups in the experiment,which were extracellular regulated protein kinases (ERK) phosphorylation inhibitor PD98059 pretreated group,Caspase-3 activation inhibitor VAD-FMK pretreated group and no PD98059 or VAD-FMK pretreated groups.And each group was divided into three sub groups with different treatment,which were adenovirus AD5-null vector infected control group,the human KAI1 gene recombinant adenovirus vector AD5 KAI1 infected group and autophagy inhibitor 3-MA pretreated and AD5-KAI1 infected group.The cell apoptosis was observed by AnnexinV-FITC/PI double staining.Caspase-3 activation level was evaluated by flow cytometry.ERK phosphorylation and poly(ADPribose) polymerase (PARP) cleavage were determined by Western blot.Results After the cancer cells infected with AD5 KAI1,KAI1 protein was expressed and GFP-LC3 green particles increased.Caspase-3 activation,PARP cleavage,ERK phosphorylation and apoptosis increased obviously.After autophagy inhibitor 3-MA pretreated,the percentage of apoptosis increased from (63.0 ± 7.9)％ to (88.0±4.5) ％ and Caspase-3 activation increased from (34.0±2.8) ％ to (44.2±4.0) ％ and PARP cleavage more.The apoptosis induced by 3-MA could be totally inhibited by Caspase-3 activation inhibitor VAD-FMK pretreated but could not be inhibited by ERK phosphorylation inhibitor PD98059.Conclusion KAI1- induced autophagy inhibits apoptosis through the downregulation of Caspase-3activation and PARP cleavage instead of ERK phosphorylation.

Objective To study the relations between the susceptibility to AIDP and AMAN, two forms of Guillain-Barr? syndrome (GBS), and the frequency of HLA-A, B alleles. Methods A case-control research was done in 31 cases of AIDP, 33 cases of AMAN and 132 health individual controls. DNA was extracted from peripheral blood leukocytes by improved fast saltingout. HLA-A, B antigens were typed by DNA-based technology, PCR-sequence specific primers (PCR-SSP) method. In determination of allelic polymorphism by PCR amplification with SSP, oligonucleotide primers are designed to obtain amplification of specific alleles or groups of alleles. Assignment of alleles is based on the presence or absence of PCR amplified product, which may be detected by agarose gel electrophoresis. Results On research of HLA-A, B alleles polymorphism, it showed that HLA-A33 frequency was increased significantly in AIDP patients [22.6% vs 4.5%,corrected probability (Pc)=0.011]; related risk (RR) was 6.1; HLA-B15, B35 frequencies were increased (51.7% vs 20.8%, Pc =0.015; 34.5% vs 6.9%, Pc=0.000 8); RR was 4.1 and 7.1, respectively. Conclusions There are different distribution of HLA-A, B alleles in AIDP and AMAN, two forms of GBS. AIDP susceptibility is associated with HLA-A33, while AMAN is with HLA-B15, B35.

Objective To observe the effect of Exendin-4 on the apoptosis of cardiomyocyte induced by H2O2,and approach the relationship between GLP-1 signal pathway and the injury of cardiomyocyte.Methods Cardiomyocytes were isolated and cultured,and were divided into 5 groups.Intercelluar ROS (reactive oxygen species) was measured,and cell apoptosis rate was evaluated by Flow cytometry in different groups.Also expressions of apoptosis-associated proteins (caspase-3) and PI3 K/AKT were evaluated by western blot.Results Compared with H2O2 group,Ex-4 co-incubation decreased the production of intercelluar ROS levels,also improved the cardiomyocyte apoptosis.At the same time,Ex-4 resulted in the alterations in expressions of the caspase-3 and p-AKT/AKT proteins.However,these effects of Exendin-4 were counteracted significantly by the co-incubation of LY294002.Conclusion The interventions of GLP-1 signal pathway can improve cardiomyocyte apoptosis induced by H2O2 incubation,and the mechanisms might partly attribute to the PI3K/AKT system.