In order to established a method for simultaneous determination of isoquercitrin, astragaline, quercetin and kaempferol in Lysimachia clethroides, the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM]PF6) methanol was used as the ultrasound-assisted extraction solvent combing with RP-HPLC. A Purospher star RP-C1 column was used with the mobile phase of aceto- nitrile, methanol and 0. 4% phosphate acid by gradient elution at the detection wavelength of 360 nm. The flow rate was 0.7 mL x min(-1), and the column temperature was the room temperature. Under the optimized conditions, the linear ranges were 2.54 x 10(-2)-2. 54, 2.50 x 10(-2)- 2.50, 1.54 x 10(-3)-0.154, 1.49 x 10(-3)-0.149 microg for isoquercitrin, astragaline, quercetin and kaempferol, respectively. The average recoveries of the four constituents were 101.1%, 98.90%, 101.0%, 101.6%, respectively. The method was green, simple, rapid and accurate, and provided a valid method for analysis of isoquercitrin, astragaline, quercetin and kaempferol in L. clethroides.
Chemical Fractionation ; instrumentation ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Flavonoids ; analysis ; isolation & purification ; Ionic Liquids ; chemistry ; Primulaceae ; chemistry

Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.
Chromatography, Reverse-Phase ; methods ; Glucosides ; analysis ; Ionic Liquids ; chemistry ; Monoterpenes ; analysis ; Oleanolic Acid ; analysis ; Paeonia ; chemistry ; Ultrasonics

Aim To observe effects of Seabuckthorn fatty acids on old rat ovarian granulosa cell apoptosis and expression of bcl-2 and fas,then discuss the adjustive effect of Seabuckthorn fatty acids on ovarian function of menopause.Methods 22 months old rats were randomly divided into 5 groups,estrogen control group,old control group,Seabuckthorn fatty acids group of high dose,middle dose and low dose.In addition,one young control group was made up of 3 months old rats.Rat ovarian granulosa cell apoptosis was detected by TUNEL.The expression of protein bcl-2 and protein fas were detected by immunohistochemistry,and bcl-2 mRNA and fas mRNA by in situ hybridization(ISH).Grey analyse was done using Q550CW image analyse system.Results Compared with the old rats control group,Seabuckthorn fatty acids of high dose,middle dose and low dose can differently restrain rat ovarian granulosa cell apoptosis,enhance the expression of protein bcl-2 and bcl-2 mRNA,weakened the expression of protein fas and fas mRNA.Conclusion Seabuckthorn fatty acids restrains ovarian granulosa cell apoptosis through adjusting the expression of bcl-2 and fas.Sequentially,it adjusts estrogen level and treats menopause syndrome.

Objective To explore the therapeutic effects of statin in patients with chronic obstructive pulmonary disease (COPD) in stable period. Methods 83 patients with COPD in stable period were recruited and randomly divided into two groups, treatment group (n = 41) and control group (n = 42).The control group received conventional routine therapy, and treatment group was given rosuvastatin calcium 20 mg once per day on the basis of routine treatment. The levels of sera C-reactive protein (CRP), pulmonary function (FEV1、FEVl%pred), and COPD assessment test (CAT) were observed at the time points of 0 week and 12th week, 24th week and 48th week were compared between two groups. Results At 12th week after the treatment, sera CRP levels, FEV1, FEV1%pred and CAT scores in either control group or treatment group showed some extent of improvements as compared to those at the time point of 0 week, and there was statistical difference of sera CRP levels and CAT scores between two groups (P < 0.05). There was no statistical difference of FEV1, FEV1%pred between two groups (P > 0.05) .24th week and 48th week after the treatment; sera CRP levels, FEV, FEV1%pred and CAT scores in two groups all significantly improved and were statistical different between two groups (P < 0.05). Conclusion Statin can alleviate inflammatory reaction, improve pulmonary function and life quality of patients with COPD.

Objective To investigate the correlation between apparent diffusion coefficient (ADC)of diffusion weighted imaging (DWI)and prognostic factors in rectal cancer.Methods 5 5 patients with rectal cancer were confirmed pathologically.Conventional pelvic MRI and DWI examination were performed,and the mean ADC values of tumor were measured preoperatively.The patients were divided into two groups with or without lymph node metastasis,and were also divided into four groups with negative,weakly positive,positive or strongly positive expression of EGFR in rectal cancer.The ADC values were calculated in each group,and the correlation of ADC values with the lymph node status and EGFR expression classification were analyzed.Results In 5 5 patients with colorectal cancer,there were 13 lesions with lymph node metastasis and the positive expression rate of EGFR was 67.2%.There were no significant difference in mean ADC value between the groups with and without lymph node metastasis (P=0.342).The number of lesions with negative,weakly positive,positive and strongly positive EGFR expression were 18,15,12 and 10.The difference in the mean ADC values among negative,weakly positive,positive and strongly positive expression groups of prognostic factor EGFR was not significantly different (P=0.412).There were also no correlations in the prognostic factors mentioned above (r=0.183 and -0.324,all P>0.05).Conclusion The ADC value can not be used to predict the prognosis and to provide more valu-able information for individualized therapy in patients with rectal carcinoma,which needs further studiy in the future.

Objective To investigate the effects of continuous positive airway pressure (CPAP) on the levels of hemorheology and C-reactive protein in patients with obstructive sleep hypopnea apnea syndrome (OSAHS). Methods 58 moderate to severe OSAHS subjects were selected as treatment group and 32 healthyadults were selected as control group. Hemorheology and C-reactive protein in all subjects were examined and compared. The levels of hemorheology and C-reactive protein were also examined and compared before and after 3 months CPAP treatment in treatment group. Results In treatment group, the levels of the whole blood viscosity and hs-CRP were significantly higher than those in control group (P < 0.01). After treatment of CPAP for three months, the levels of the whole blood viscosity and hs-CRP obviously decreased (P < 0.01). Conclusion Levels of hemorheology and hs-CRP are elevated in patients with OSAHS and CPAP therapy could effectively decrease serum levels of hemorheology and hs-CRP in patients with OSAHS.

Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P ＜0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.

Objective To study the effect of self-efficacy on learning effect of medical students.Methods Self-efficacy scores of the questionnaire to clinical medical students who practice in our hospital from September 2012 to April 2013 were measured.Students with score lower than the average were recruited into group A,and the others were recruited into group B.Then the scores of both-group students in the process of clinical practice,the theoretical knowledge,and triguaiacyl tests were gotten for statistical analysis.Results Group B (88.02 ± 3.902; 89.02 ± 3.902; 89.94 ± 3.794) were significantly better than group A (67.07 ± 5.131,66.02± 5.866,65.23 ±6.741) in the scores of the theory of knowledge,operation skills and triguaiacyl test.The correlation analysis showed that self-efficacy was positively related to these three aspects (P=0.032,0.024,0.001).Conclusion Clinical teachers should consciously improve students' self-efficacy in the process of their clinical practice,in order to improve the learning effect of the clinical medical students preferably.

Objective To explore the correlation among serum total bilirubin (TBil) , invasive hemodynamic parameters, plasma N-terminal proBNP (NT-preBNP) and C reactive protain (CRP)in patients with heart failure. Methods Invasive hemodynamic parameters derived from Swan-Ganz catheter, TBil, plasma NT-proBNP and CRP within 12 hours after hospital admission were analyzed in 130 patients with chronic heart failure [Now York Heart Association (NYHA) class Ⅱ -Ⅳ]. Results Compared with those in non-hyperbilirubinemia group, pulmonary capillary wedge pressure (PCWP), NT-proBNP and left ventriculure ejection fraction were different significantly in total hyperbilirubinemia group [(26. 09 vs 16.00) mm Hg(1 mm Hg =0. 133 kPa), (3.36 vs2.91) pmol/L, (34. 12 vs 28.92)%, P<0. 05]. The serum TBil increased significantly in higher PCWP, right atrial pressure and NT-proBNP groups than those in lower level groups [(32. 22 vs 24. 17), (37.52 vs 24. 19), (32. 14 vs 16. 74) pmol/L, P < 0.05]. Partial correlation analysis showed serum TBil was associated with PCWP, right atrial pressure, pulmonary vascular resistance index and NT-proBNP respectively (r = 0. 21, P = 0. 02; r = 0. 33, P < 0. 01 ; r = 0. 20, P =0. 04;r = 0. 37, P <0. 01, respectively). Multiple linear regression analysis showed both right atrial pressure and NT-proBNP correlated independently with serum TBil(β= 0. 39, P < 0. 01 ;β = 0. 29,P = 0. 01, respectively). Conclusion For patients with heart failure, serum TBil correlated well with right atrial pressure, PCWP and NT-proBNP; it is a reliable indicator for exact clinical evaluation of heart failure.

To investigate the metabolic rate and metabolites of 9-dehydro-17-dehydro-andrographolide, which is the main active ingredient in Xiyanping injection, by using the in vitro rat liver microsome incubation system. 9-dehydro-17-dehydro-andrographolide was incubated together with liver microsome mixed with NADPH. Its metabolic rate was studied by determining its residual concentrations with the UHPLC-MS/MS method; Its metabolites were identified by the UPLC-TOF-MS(E) method. The results showed that 9-dehydro-17-dehydro-andrographolide was metabolized faster than rat liver microsomes mixed with coenzymes, with t½ and CL of (19.7 ± 0.5) min and (35.1 ± 0.8) mL x min(-1) x g(-1) (protein), respectively. Based on the high resolution mass spectrum data and information from literatures, altogether nine metabolites of 9-dehydro-17-dehydro-andrographolide were identified in the incubation system, particularly hydroxylated and dehydrogenized products. The results of identification would provide a basis for screening out more active andrographolide derivatives.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Microsomes, Liver ; chemistry ; metabolism ; Molecular Structure ; Rats ; Tandem Mass Spectrometry