[Objective]To establish a method to determine the contents of glucase and fructose in extracts of leaves of Lonicera hypoglauca Miq. using HPLC-ELSD, and apply this method to study the content change of glucose and fructose in different solvents extracts in leaves of Lonicera hypoglauca Miq. [Methods] The chromatographic separation was achieved on Lichrospher NH2 column using a mobile phase composed of a mixture of acetonitrile and water(90:10) at a flow rate of 0.8mL·min-1 and the column temperature was 25℃.The ELSD drift tube temperature was set at 70℃ and the carrying gas was N2 and the flow rate of the gas was 1.6L·min-1. [Results] Glucose and fructose from extracts of the leaves of Lonicera hypoglauca Miq. could be wel separated, and the linearity ranges of glucose and fructose were shown at the concentration of 0.5840~1.168mg·mL-1 and 0.8256~1.8576mg·mL-1, the average recovery rate of glucose and fructose were 96.88%(RSD=1.11%) and 97.55%(RSD=1.15%), the stability of glucose and fructose was 2.12%and 0.38%, and the repeatability was 1.31% and 0.24%. Among the water extract,50% alcohol extract and 95% alcohol extract,the content of the glucose and fructose in the water extract was highest,which was 2.19mg·mL and 4.10mg·mL-1, the 50% alcohol extract was second, which was 1.13mg·mL-1 and 2.12mg·mL-1, and the 95% alcohol extract was lowest, which was 0.45mg·mL-1 and 0.65mg·mL-1.[Conclusion] The method is simple, accurate and has good repeatability, and can be used to determine the contents of glucose and fructose in the extract of leaves of Lonicera hypoglauca Miq. In this batch of leaves of Lonicera hypoglauca Miq., the contents of fructose and glucose were of water extract>50%ethanol extract>95%ethanol extract.

Objective To compare the effects of equivalent dose sufentanil,remifentanil or fentanyl on hemodynamic and cardiac output in the elderly patients during induction of general anesthesia.Methods Ninety elderly patients(ASA Ⅰ ～ Ⅱ) undergoing elective abdominal surgery under general anesthesia were randomly divided into sufentanil group(group S),remifentanil group(group R) and fentanyl group(group F),30 patients in each group.Patients in group S,group R and group F received sufentanil 0.2μg/kg,remifentanil 2μg/kg and fentanyl 2μg/kg Ⅳ,respectively,using an blind method before intubation.SBP,DBP,MAP,HR,CO,CI and SVR were recorded before of anesthesia (T0),immediately after induction (T1),1,3 minutes after tracheal intubation (T2,T3).Changes of SBP and HR during observation were also recorded.Results The MAP and HR in three groups at T1 were significantly lower than those at T0 and decreased significantly in group R than those in group F and S(P ＜0.05).The MAP and HR in group F significantly increased at T1 than T0.The MAP and HR in group R were significantly decreased after tracheal intubation.Compared with the baseline,the MAP and HR in group S at T1,T2,remained unchanged.In group R,there were two patients whose HR were under 50 bpm.In group F,the MAP at T1,T2 were significantly higher than those at T.In group S,there were no significant changes in CO,CI,SVR which were all within normal range (P ＞ 0.05).Conclusion Both sufentanil and remifentanil effectively inhibit the stress response during induction of general anesthesia.At the same time,sufentanil has better hemodynamic stability.

Objective To collect data and analyze the current status and prevalence changes of Crohn's disease (CD) in mainland China in recent decades. Methods A computer-based literature search was previously performed by using 50-year (1950 to 2002) of records of CD from the Chinese Database of Biology and Medicine (CBM) (1979 to 2002) and a manual year-by-year search of the literature ( 1950 to 1978). Using similar method, descriptive epidemiological data from 2003 to 2007 were collected, analyzed and compared with previous research. Results Four hundred and seventeen relevant papers during 2003 and 2007 were collected and 62 papers were eligible for inclusion. Within 62 papers, a total of 2149 cases with CD from 2003 to 2007 have been reported nationwide, comprising 1288 male and 861 female patients, result (ie,1526 cases from 1950-2002). There were no obvious changes in incidence age (younger and middle age were main components) and sex ratio ( number of male was still larger than that of female). The extrapolated CD incidence and prevalence rates were 1.21/100 000 person ? year and 2.29/100 000, respectively, which were higher than that of year 1950-2002, 0.28/100 000 person ? year and 1.38/ 100 000, respectively. Conclusions The incidence and prevalence rates of CD have been increasing rapidly, but these rates are still lower than those in Western world.

Objective To investigate the effect of ischemia and ischemia/reperfusion(I/R)in rat heart on matrix metalloproteinase-1(MMP-1).Methods The I/R animal models were established by shutting down and reopening the anterior interventricular branch with a silver clamp,then the distribution and amount of MMP-1 of the normal and I/R rat hearts were observed by immunohistochemical staining and Western blotting and analyzed by computer image analysis.Results 1.Immunohistochemical staining showed MMP-1 existed mainly in the cardiac matrix.There were strong positive reactions in fibrocytes,smooth muscle cells of the blood vessel and endotheliaI cells of capillaries.MMP-1 didn't show distinct changes 30 minutes after ischemia,while its concentration increased dramatically 60 minutes after ischemia.The positive reaction of MMP-1 increased 30 minutes after I/R,and 60 minutes after I/R there was large fusion areas in MMP-1 existing reglons.2.Quantitative analysis showed no dramatic changes of MMP-1 after ischemia for 30 minutes(P＞0.05),while dramatic changes were seen 60 minutes after ischemia(P＜0.05).MMP-1 changed dramatically 30 minutes and 60 minutes after I/R.3.Western blotting showed that there were no distinct naked-eye-observable changes.The bands of MMP-1 became widened 30 minutes after I/R,and became obviously widened 60 minutes after I/R.Conclusion 1.MMP-1 is secreted by fibrocytes,smooth muscle cells and endothelial cells of cardiac tissue under physiological conditions,and cardiomyocytes has the potential to secrete MMP-1 under ischemia or I/R.2.The longer time the heart ischemia lasts,the greater MMP-1 concentration will increase.Reperfusion can increase MMP-1 concentration to an even higher level,which may be the main cause of the collagen destruction after heart I/R.

Background and purpose: Reports showed that berbefine not only had an effect ofdetoxification and inflammation but also could induce the cell to apoptosis. Berberine may also inhibit the migration and metastasis of tumor. To investigate the effects and mechanism of berberine on migration and metastasis of A549 cells, the proliferation, mobility and adhesion of A549 cells were observed after incubation with berberine. Methods: The proliferation was determined by MTT. Wound healing assay and adhesion test were used to observe the mobility and adhesion rate. The mRNA and protein expressions of MMP2 and MMP9 were assessed by semi-quantitive RT-PCR and gelatin zymography, respectively. Results: Berberine could significantly inhibit the proliferation ofA549 within a certain range of treating time and dose (P<0.05). The mRNA and protein expression of MMP2 and MMP9 gene were decreased slightly by the treatment ofberberine (P<0.05), especially at dose of 100 μg/ml. Conclusion: Berberine can inhibit the migration and adhesion abilities of A549 cells, probably by down-regulating the expression and activation of MMP2 and MMP9.

To evaluate the application of intracavitary three-dimensional ultrasonography in the diagnosis of polycystic ovary syndrome ( PCOS). The three-dimensional volumes of ovary were measured by intracavitary ultrasonography in 40 PCOS patients and 40 normal controls. The diagnostic value of parameters derived from three-dimensional ultrasonography was evaluated by using the receiver operating characteristic (ROC) curve. The ovarian volume, stromal volume, and the stromal/total volume ratio in PCOS group were significantly higher than those in control group (both P ＜ 0. 05 ) ; the area under ROC curve ( AUC ) of them were 0. 876, 0. 926, 0. 886 respectively. Among these parameters, stromal volume had the highest diagnostic value for PCOS, the cut-off value of which was 7.52 cm3 with a sensitivity of 88% and a specificity of 90%. Volume parameters of ovary measured by intracavitary three-dimensional ultrasonography can offer quantitative information for the diagnosis of PCOS.

Objective To investigate the safety and clinical response of dendritic cells (DC)vaccine loaded with HLA-A2-restricted peptides MAGE-3、 Tyr、 MART-1 and GP-100 against malignant melanoma. Methods Twenty three HLA-A2 positive patients with malignant melanoma were enrolled.Peripheral blood mononuclear cells were separated into adherent cells for induction of DC loaded with four peptides. DC vaccine was administered subcutaneously once a week in inguinal region. The immunological responses and clinical responses were evaluated. Results Dendritic cell vaccination were well tolerated in all patients and there was no toxicity observed. Serum levels of IL-2, IL-12 and IFN-γincreased significantly after first vaccination and fourth vaccination. The positive rate of DTH test was 50% (5/10), and 2 fold increase of CD8+ IFN-γ+ cells were observed in 6 of 10 patients. Stable disease was observed in 11 of 23 patients, one patient had a complete metastasis regression, four patients had 50% regression of metastasis,four patients had a minor response, and disease progressed in three patients. Conclusion DC vaccines loaded with peptides MAGE-3、 Tyr、 MART-1 and GP-100 can elicit non-specific and specific immune responses, leading disease control. DC vaccine is considered one of safe and effective treatments for malignant melanoma.

Objective To study emptying of gallbladder in children with functional dyspepsia(FD),and to investigate correlation of their helicobacter pylori(Hp) status and gallbladder emptying rate.Methods Sixty children with FD were studied,including 30 Hp-ne-(gative) and 30 Hp-positive children.Thirty Hp-positive children received triple eradication therapy 1 week.After 4 weeks,the Hp tests were taken again,27 children became Hp-negative,and the others were still positive eliminated from the study.The difference between the above groups in gallbladder volumes before breakfast and postprandial gallbladder emptying rate were compared.Results The gallbladder volumes before breakfast were bigger and the emptying rate of postprandial gallbladder of children with FD were lower than those of normal controls(P0.05).Conclusions Delayed emptying of gallbladder may cause FD in children.Hp infection isn′t associated with the gallbladder emptying rate.

The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; metabolism ; Glucosides ; metabolism ; Hydrolysis ; Mass Spectrometry ; Metabolic Networks and Pathways ; Pinaceae ; chemistry ; Rats

For effective inhalable dry-powder drug delivery, tetrandrine-PLGA (polylactic-co-glycolic acid) nanocomposite particles have been developed to overcome the disadvantages of nanoparticles and microparticles. The primary nanoparticles were prepared by using premix membrane emulsification method. To prepare second particles, they were spray dried. The final particles were characterized by scanning electron microscopy (SEM), dry laser particle size analysis, high performance liquid chromatography (HPLC), X-ray diffraction (XRD), differential scanning calorimetry (DSC), infrared analysis (IR) and confocal laser scanning microscope (CLSM). The average size of the primary particles was (337.5 ± 6.2) nm, while that second particles was (3.675 ± 0.16) μm which can be decomposed into primary nanoparticles in water. And the second particles were solid sphere-like with the drug dispersed as armorphous form in them. It is a reference for components delivery to lung in a new form.