Objective To establish an HPLC method for determination of simvastatin and its related substances.Methods The HPLC system consisted Luna C18 column(250mm×4.6mm,5μm),the mobile phase was 0.025mol/L sodium phosphate monobasic(pH 4.5):acetonitrile(35:65).The flow rate wns 1.0ml/min,and with detection 238nm.The column temperature was room temperature;injeetion volume WaS 20μl.Results SimvaStatin and its related substances can be completely separated by the method with a linear range of 2～200μg/ml.Calibration culwe was linear over the range of Y=3.256C×103+4.510×102(r=0.9996);for potency assay the mean recovery rate was 99.32% with RSD as 0.31%(n=9).Conclusion The method has good reproducibility,high accuracy and strong specificity.It was suitable for determination of simvastatin and its related substances.

A high performance liquid chromatography-inductively coupled plasma mass spectrometric ( HPLC-ICP-MS) method for analysis of selenocystine (SeCys2), selenite (SeⅣ), selenomethionine (SeMet) and selenate ( SeⅥ) was described. Chromatographic separation was performed using 8 mmol/L citric acid solution (pH 5). The potentially interfering 40Ar40Ar+ at selenium masses of m/z 80 was reduced using CH4 as a reactive cell gas in the dynamic reaction cell. The selenium species was extracted using ultrasonic extraction system with a mixture of protease K and lipase. The limits of detection for SeCys2 , SeⅣ, SeMet, and SeⅥwere observed as 0. 088, 0. 033, 0. 30, and 0. 17 ng/mL, respectively. The correlation coefficients were greater than 0 . 9995 . The recoveries were observed in the range of 95 . 1%-114 . 6%. The developed method was successfully applied for the determination of selenium species in Burdock and Panax notoginseng.

To study the chemical constituents of the stems of Clematis parviloba, six compounds were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Two new phenolic glycosides, 2-((E)-3-carboxybut-2-en-yl)-4-hydroxy-3-methyl-phenyl-O-beta-D-glucopyranoside (1) and 4'-hydroxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (2) were isolated, together with a known phenolic glycoside, 4'-hydroxy-3'-methoxy-phenol-beta-D-[6-O-(4"-hydroxy-3", 5"-dimethoxy-benzoate)] glucopyranoside (3) as well as three known megastigmane glycosides, linarionoside A (4), linarionoside C (5), and staphylionoside K (6). Their structures were determined on the basis of spectroscopic analysis and chemical evidence. Among them, compounds 1 and 2 were named as clemaparvilosides A (1) and B (2), respectively, and compounds 3-6 were obtained from Clematis genus for the first time.

lusions Cardiac MSCT can accurately assess the RV size and function in comparison to MRI. Patients with severe COPD have RV dysfunction.

Objective To prospectively assess right ventricular dysfunction and the metergasis before and after therapy of thrombolysis with ECG gated multi-detector spiral computed tomography (MSCT)in patients with acute pulmonary embolism. Methods Triple rule-out ECG gated MSCT examination was performed in 96 consecutive patients suspected of PE. 25 patients with central PE were confirmed. 25 agematched subjects without cardiac and pulmonary disease were recruited as control group. Triple rule-out ECG gated MSCT were performed again to assess cardiac function after therapy of thrombolysis. Dimension ratios for the right ventricle (RV) and left ventricle ( LV), main pulmonary artery and aorta were measured.Furthermore, the RV and LV end-diastolic volumes (EDV), end-systolic volume (ESV) and ejection fraction (EF) were also measured. The mean values were compared with analysis of variance (ANOVA) and Newman-Keuls test before and after thrombolysis. Results The mean values of RVEDV, RVESV, RVEF,RV/LV ESV volume ratio, RV/LV dimension ratio and main pulmonary artery/aorta dimension ratio in control group were (150.5±24.1) ml,(71.5 ±18.5) ml, (53.5 ±4.2)%, 1.08 ±0.04, 1.01 ±0.04 and 0. 99 ±0. 02, respectively. While those in PE group were ( 190. 3 ± 16. 2) ml, ( 128. 1 ± 13.2) ml,(32.7 ± 3.6 ) %, 2.00 ± 0.26, 1.30 ± 0. 09 and 1.34 ± 0. 11, respectively. Those after therapy of thrombolysis were ( 159. 2 ± 21.5 ) ml, ( 80. 7 ± 9.4) ml, (49. 2 ± 5.9) %, 1.22 ± 0.25, 1.02 ± 0.02 and 1.02±0.11,respectively. ESV and EDV of RV were larger (q= 6.28, P<0.01; q=7.59, P<0.01),EF value was lower (q = 4. 82, P < 0.01 ) in PE group than those in control group. RV/LV ESV volume ratio, the RV/LV dimension ratio and main pulmonary artery/aorta dimension ratio were larger ( q = 6. 04,P <0. 01; q =4. 43, P <0. 01; q =4. 36, P <0. 01 ) and EDV of LV was lower in PE group than those in control group. However, ESV and EDV of RV and RV/LV ESV volume ratio were lower (q = 5.03, P <0.01;q=6. 11,P<0.01;q=4.74,P<0.01), EF value was larger (q=6.29, P<0.01) and EDV of LV was larger(q =4.01 ,P <0.01 ) after therapy of thrombolysis than before. Conclusion Retrospective triple rule-out ECG-gated MSCT can show pulmonary embolism, measure the function of RV and LV and evaluate curative effect of thrombolysis.

BACKGROUND:There is a group of leukemic stem cell (LSC) in patients with leukemia. These malignant stern cells, although rare, but self-renewal, with a certain degree of differentiation is the existence of leukemic cells and causes of their continued proliferation. Intervention of leukemia-specific targeted treatment drugs has become a research hotspot in recent years. OBJECTIVE: To investigate the influence of traditional medicine compound on expression of flt3 and N-ras mRNA in LSCs. DESIGN, TIME AND SETTING: A randomized, grouping experiment was performed at the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from September 2007 to December 2008.PARTICIPANTS: A total of 50 cases of acute myelocytic leukemia admitted to Department of Hematology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine and Qilu Hospital of Shandong University between 2006 and 2007 were selected, including 5 cases of type FAB, 15 of M2, 9 of M4 and 21 of M5.METHODS: Original prescription: Huangqi, Hedyotic diffusa, Xiaoji, Taizishen, Banzhilian, Pugongying, Rehmannia dride rhizome, Huangjing, Nuzhenzi, ecliptae herba, Tiandong, Maidong, Baizhu, Fuling, Liquorice root. Fuzheng prescription: Huangqi, Hedyotic diffusa, Xiaoji, Banzhilian, Pugongying, ecliptae herba. Preparation: the above-mentioned three groups of drugs were prepared into 1 g/mL medicine liquid by Laboratory of Chinese Drug Preparation of Affiliated Hospital of Shandong University of Traditional Chinese Medicine. When it was negative in sterility test, the liquid was filtrated and sterilized. 5 mL bone marrow was separately harvested from M1, M2, M4, and M5 leukemia patients, diluted and placed in lymphocyte isolation solution. LSCs were purified by magnetic activated cell sorting and flow cytometry. The cell concentration was adjusted to 2×108/L and divided into 4 groups: control (no treatment), and three experimental groups (treated separately with original, Fuzheng, Quxie prescriptions at final concentration of 100 mg/L). The cells were cultured for 48 additional hours. MAIN OUTCOME MEASURES: RT-PCR was used to detect the mRNA expressions of Flt3 and N-ras in LSCs. RESULTS: The expression of flt3 in three experimental groups was significantly decreased compared with control group (P < 0.05), in particular, original prescription group decreased the most (P<0.05), and no significant difference was found between Fuzheng and the Quxie prescription groups (P>0.05). The expression of N-ras in four groups was similar to Flt3. CONCLUSION: Flt3 and N-ras mRNA were overexpressed in LSCs. Chinese medicine original prescription and its Fuzheng or Quxie prescriptions decreased Flt3 and N-ras mRNA expression.

Objective To investigate the effect of insulin and gliclazide therapy on endoplasmic reticulum (ER) stress and insulin sensitivity in the liver of type 2 diabetic rats.Methods A high fat diet plus a low-dose of streptozotocin was implemented to create a type 2 diabetic rats which were randomly divided into diabetes mellitus (DM) group,insulin treatment (INS) group and gliclazide treatment (GT)group; and healthy rats were as normal control group.Diabetic rats in INS and GT groups were given neutral protamine hagedorn (NPH) insulin and gliclazide respectively for 3 weeks.Protein expression levels of immunoglobulin binding protein (Bip),spliced X-box binding protein 1 (XBP-ls),phosphorylated c-Jun on serine 73 (p-c-Jun),phosphorylated insulin receptor substrate 1 on serine 307 (p-IRS-1),and glucose-6-phosphatase (G6Pase) in liver homogenate were detected by Western blotting.Results Compared with the normal rats,Bip and XBP-Is in the DM group were up-regulated (0.28 ±0.07 vs 0.90 ±0.10 for Bip;0.41 ± 0.07 vs 0.95 ±0.07 for XBP-1 s; both P ＜ 0.01 ) ; p-c-Jun (0.59 ± 0.18 vs 1.94 ± 0.03 ),p-IRS-1( 1.73 ± 0.18 vs 5.32 ± 0.22) and G6Pase (0.11 ± 0.01 vs 0.45 ± 0.01 ) were increased ( all P values ＜0.01 ).In the INS group,all of aforementioned changes were reversed (0.90 ± 0.10 vs 0.25 ± 0.04 for Bip; 0.95 ±0.07 vs 0.47 ±0.01 for XBP-1s; 1.94 ± 0.03 vs 0.50 ±0.10 for p-c-Jun; 5.32 ± 0.22 vs 1.59 ±0.32 for p-IRS-1 ; 0.45 ±0.01 vs 0.15 ±0.02 for G6Pase,all P values ＜0.01 ).In the GT group,all of aforementioned changes were also attenuated ( 0.90 ± 0.10 vs 0.53 ± 00.02 for Bip ; 0.95 ± 0.07 vs 0.78±0.02 for XBP-1s; 1.94 ±0.03 vs 1.33 ±0.11 for p-c-Jun; 5.32 ±0.22 vs 3.13 ±0.02 for p-IRS-1; 0.45 ± 0.01 vs 0.25 ± 0.01 for G6Pase,all P values ＜ 0.05).Furthermore,all of aforementioned protein levels were down-regulated more obviously in the INS group comparing to the GT group ( all P values ＜ 0.01 ).Conclusions Both insulin and gliclazide therapy could relieve ER stress and e-Jun N-terminal kinase activity and improved insulin sensitivity.The effect of insulin on Bip,XBP-1s,p-c-Jun,p-IRS-1 and G6Pase protein expressions is more obvious than that of glilcazide,which indicates besides lowering glucose,insulin might have protective effects of anti-inflammation,anti-oxidative stress or stimulation of lipid redistribution.

Most of the maxillary first premolar are single-rooted,there are also two-rooted and three-rooted ones.A case of three-rooted maxillary first premolar was diagnosed by CBCT.CBCT has the advantage in the observation of the variation root number,root canal bending and the variation of root canal morphology.

Objective To investigate the impact of polyclonal neural cell adhesion molecule antibody (P-NCAM-Ab) on the potency of botulinum toxin A (BTX-A).Methods Ninety male Sprague-Dawley rats were randomly divided into 3 equal groups:a normal control group,a BTX-A group and a P-NCAM-Ab group.The rats in the normal control group were injected with 100 μl of saline solution in their right gastrocnemius,while those in the BTX-A and P-NCAM-Ab groups were injected with 100 μl of BTX-A (0.5 U).In addition,the rats in the P-NCAM-Ab group were also injected with 100 μl of P-NCAM-Ab (the dosage was 20 U) at the same site on the 3rd day after the BTX-A injection.The rats' gastrocnemius muscle strength was evaluated with a self-made system for evaluating neuromuscular function before and after the toxin injection,on the 3rd day,as well as 1,2,4,6,8,10 and 12 weeks after the BTX-A injection.Any wet weight changes in the muscles were observed,and immunochemistry methods were employed to observe any structural changes in the motor endplates and nerve fibers at the different time points.Results After the saline injection,the average gastrocnemius muscle strength of the control group increased with time,while strength in the BTX-A and P-NCAM-Ab groups demonstrated a decrease in strength followed by a gradual increase.The average gastrocnemius muscle strength of the rats in the BTX-A and P-NCAM-Ab groups was significantly lower than that of the control group at all time points.Compared with the BTX-A group,the muscle strength of the P-NCAM-Ab group rats decreased further.Strength recovery in the BTX-A and P-NCAM-Ab groups was significantly slower than in the control group.The wet weight percentage in the BTX-A and P-NCAM-Ab groups at first decreased and then recovered with time.After the BTX-A injection,the average wet weight percentage of the P-NCAM-Ab group rats was significantly lower than that of the BTX-A group after 3 days,and 1,2 and 4 weeks.Karnovsky-Roots AchE staining showed that the motor endplates' color in the BTX-A and P-NCAM-Ab groups deepened gradually,though the color of the P-NCAM-Ab group was lighter than that of the BTX-A group at each time point.The mean optical density of the motor endplates' positive reaction area increased with time in both groups,but the P-NCAM-Ab group was lower than that of the BTX-A group at 1,2,4,8 and 12 weeks.Counting the nerve fibers dyed by gold chloride showed similar trends with both experimental groups significantly different from the control group.Conclusion P-NCAM-Ab can increase the potency of BTX-A and prolong its action.