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

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

OBJECTIVETo explore the value of spinal canal and dural sac dimensions for the treatment of lumbar disc herniation in MRI.

METHODSThe clinical data of 144 patients with single-level lumbar disc herniation underwent nonsurgical or surgical treatment from January 2010 to December 2012 were retrospectively analyzed. There were 91 patients in the nonsurgical group, including 55 males and 36 females, ranging in age from 20 to 68 years old with an average of (43.37±12.48) years; and there were 53 patients in the surgical group, including 28 males and 25 females, ranging in age from 20 to 64 years old with an average of (42.98±12.95) years. JOA scores (29 scores) were used to evaluate clinical manifestation (including subjective symptoms, objective findings, limitation of daily activities and bladder function) and outcomes. The parameters related to spinal canal and dural sac dimensions (including spinal canal midsagittal diameter and available diameter, lateral recess width, spinal canal and dural sac cross-sectional area) in the initial axial T2-weighted MRI were measured, and odds ratio of available diameter to midsagittal diameter, odds ratio of lateral recess width to midsagittal diameter and area ratio of dural sac to spinal canal were calculated. Then, the differences of all parameters between two groups, and the correlations with initial JOA scores were analyzed.

RESULTS(1) All patients were followed up from 1 to 3 years with an average of 2.1 years. JOA scores before treatment were 16.27±2.96 in nonsurgical group and 12.64±3.30 in surgical group, there was statistically significant difference (t=6.319, P<0.01). At final follow-up time, there was no statistically significant difference in JOA scores (25.41±2.22 vs 25.76±2.29), improvement rate [(72.95±12.54)% vs (76.80±9.45)%], and the excellent and good rate (84.91% vs 78.02%) between two groups (P>0.05). But, the relapse rate of nonsurgical group was higher than surgical group (14.29% vs 5.67%). (2) Spinal canal midsagittal diameter and available diameter, lateral recess width, spinal canal and dural sac area, the ratio of available diameter to midsagittal diameter, and the ratio of lateral recess width to midsagittal diameter in surgical group were smaller than that of nonsurgical group, but the area ratio of dural sac to spinal canal was larger, and there were statistically significant differences between two groups (P<0.01). (3) The initial JOA scores showed significantly positive correlation with spinal canal midsagittal diameter and available diameter, lateral recess width, and canal and dural sac area (P<0.01); also presented positive correlation with the ratio of available diameter to midsagittal diameter and the ratio of lateral recess width to midsagittal diameter (P<0.05); but there was a significantly negative correlation between initial JOA scores and the area ratio of dural sac to spinal canal.

CONCLUSIONBoth nonsurgical and surgical treatment of lumbar disc herniation can obtain good effect, but the recurrence rate of non-surgical treatment is higher. Preoperative MRI measurement parameters of spinal canal and dural sac dimensions has certain value for the treatment selection of lumbar disc herniation, but further refinement and validation is still required.

Adult ; Aged ; Dura Mater ; pathology ; Female ; Humans ; Intervertebral Disc Displacement ; pathology ; therapy ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Spinal Canal ; pathology

Objective:To analyze the reflux parameters of patients with gastroesophageal reflux disease (GERD) in upright position, supine position and at 2 h after meals, and to explore the cut-off value, sensitivity and specificity of the reflux parameters in different positions and at 2 h after meals in GERD diagnosis.Methods:From January 2016 to July 2020, 200 GERD patients (GERD group) and 61 non-GERD patients (control group) who visited Huazhong University of Science and Technology Union Shenzhen Hospital (Former Nanshan District People′s Hospital), were selected. All the patients of the two groups received gastroesophageal reflux disease questionnaire (GERDQ), upper gastrointestinal endoscopy, esophageal high resolution manometry and 24 h esophageal pH combined impedance monitoring. T test, non-parametric test and chi-square test were used to compare the related parameters in upright position, supine position and at 2 h after meals between two groups and within each group. Receiver oparative characteristic (ROC) curves of reflux parameters in upright position, supine position and 2 h after meals were drawn to determine the cut-off value, sensitivity and specificity in GERD diagnosis. Results:The proportion of patients with acid reflux in supine position of the control group was higher than that of the GERD group (41.0%, 25/61 vs. 8.50%, 17/200), and the difference was statistically significant ( χ2=36.53, P<0.01). In the control group, the acid reflux time in upright position, number of acid reflux, acid exposure time (AET), longest reflux time and number of weak acid reflux were more than those of in supine position in the same group (6.00 min(2.00 min, 13.50 min) vs. 0.00 min(0.00 min, 1.50 min), 16.00(8.00, 27.00) vs. 1.00(0.00, 3.00), 0.90%(0.33%, 1.88%) vs. 0.00%(0.00%, 0.30%), 2.00 min(1.00 min, 4.00 min) vs. 0.00 min(0.00 min, 1.00 min), 7.00(3.00, 11.00) vs. 1.00(0.00, 2.00), respectively) and the differences were statistically significant ( Z=5.43, 6.61, 5.06, 3.58 and 6.24, all P<0.01). In the GERD group, the acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux in upright position were higher than those in supine position (51.00 min, (31.00 min, 86.75 min) vs. 8.00 min(1.00 min, 42.00 min), 60.00(48.00, 83.75) vs.6.00(2.00, 19.50), 7.30%(3.90%, 12.10%) vs. 1.50%(0.20%, 6.50%), 7.00 min(4.00, 12.00 min) vs. 4.00 min(1.00 min, 17.00 min), 1.00(0.00, 3.00) vs. 0.00(0.00, 2.00), 7.00(3.00, 12.00) vs. 0.00(0.00, 1.00), respectively) and the differences were statistically significant ( Z=7.92, 11.22, 6.90, 2.56, 5.11 and 11.76, all P<0.05). The acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux at 2 h postprandial were 3.00 min(2.00 min, 9.00 min), 10.00(5.00, 18.00), 0.90%(0.40%, 1.98%), 1.00 min(0.00 min, 3.00 min), 4.00(1.50, 8.50)and 28.50 min(15.00 min, 54.75 min), 35.00(24.00, 52.00), 8.30%(4.32%, 15.83%), 6.00 min(3.00 min, 11.00 min), 4.00(2.00, 7.25), in the control and GERD groups, respectively, which were significantly higher than those in supine position in the same group ( Z=4.30, 6.33, 5.50, 3.40, 5.71 and 3.76, 9.21, 5.76, 1.97, 10.46, all P<0.05). Among 200 GERD patients, 125 patients had symptoms recorded during the 24 h esophageal pH combined impedance monitoring, the incidence of reflux symptoms in upright position was higher than that in supine position (89.6%, 112/125 vs. 65.6%, 82/125), and the difference was statistically significant ( χ2=20.71, P<0.01). The results of ROC curve analysis showed that the accuracy of acid reflux time in upright position in GERD prediction was the highest, with AUC value of 0.94 and cut-off value of 24.5 min, and the sensitivity and specificity in GERD diagnosis were 81.50% and 95.08%, respectively. The prediction accuracy of acid reflux times in upright position and AET in upright position for GERD was secondary, AUC value both were 0.93 and the cut-off value of the acid reflux number in upright position was 39.5, and the sensitivity and specificity in GERD diagnosis were 84.00% and 95.08%, respectively. The cut-off value of AET in upright position was 2.75%, the sensitivity and specificity in GERD diagnosis were 85.00% and 93.33%, respectively. The AUC value, cut-off value, sensitivity and specificity of AET at 2 h postprandial were 0.91, 4.60%, and 73.49% and 95.00%, respectively. Conclusions:Both GERD patients and non-GERD patients have more reflux in upright position, especially within 2 h after meals. The diagnostic values of acid reflux time in upright position, number of acid reflux, AET and AET 2 h after meals for GERD is high, and the AUC values are all >0.90, which can be used as a more comprehensive basis for the analysis and diagnosis of GERD.

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 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 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.