11.0 mmol/L).Perioperative decontamination in digestive tract was a protective factor in the prevention of bacterial infection.Conclusion Bacterial infection is one of the most severe complications after OLT.Therefore,it is very important to remove those risk factors,make early diag- nosis and take effective treatment.

High-speed counter-current chromatography (HSCCC) was used to high performance separate and prepare lignans from Schisandrae chinensis fructus. The solvent system is composed of n-hexane-ethyl acetate-methanol-water (9 : 1 : 5 : 5) and n-hexane-ethyl acetate-methanol-water (9 : 1 : 9 : 5), speed is at 900 r.min-1, and flow rate is at 2.0 mL.min-1. Five fractions from Schisandrae chinensis fructus extract were separated and prepared with one HSCCC process. They were identified as schisandrin, gomisin J, schisandrol B, schisantherin A and deoxyschizandrin by electrospray ionization-multiple tandem mass spectrometry (ESI-MSn), respectively. Their contents were obtained in 98.74%, 94.32%, 99.53%, 94.23% and 98.68% by ultra high performance liquid chromatography (UPLC), separately. The rapid and simple method can be applied for the preparation of lignans from Schisandrae chinensis fructus.
Countercurrent Distribution ; Cyclooctanes ; chemistry ; isolation & purification ; Dioxoles ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fruit ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; chemistry ; isolation & purification ; Schisandra ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

In order to affirm the cardioactive components in Fuzi, we identified a group of aminoalcohol- diterpenoid alkaloids in Fuzi using ultra high-performance liquid chromatography coupled with electrospray ionization mass spectrometer (UPLC-ESI-MS) method. Among a total of forty-one isolated ingredients, thirteen major aminoalcohol-diterpenoid alkaloids were identified by comparing their retention times and MS spectra with those of the reference substances. Moreover, Fuzi samples from different places of origin and with different processing methods were examined and their components displayed a pattern of high similarity, though the relative abundance varies probably due to their different processing methods. Furthermore, the cardiac effect of each identified alkaloid was individually evaluated using the isolated bullfrog heart perfusion experiment. Among the thirteen aminoalcohol diterpenoid alkaloids tested, six of them significantly enhanced the amplitude rates. Taken together, we affirm that the cardioactive components in Fuzi are aminoalcohol-diterpenoid alkaloids, shedding light on future studies of the mechanisms and development of these cardioactive compounds.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Amino Alcohols ; chemistry ; Animals ; Cardiotonic Agents ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Heart ; drug effects ; In Vitro Techniques ; Plant Extracts ; chemistry ; Rana catesbeiana ; Spectrometry, Mass, Electrospray Ionization

The purpose of this study was to evaluate the impact of callus induction and culture conditions on secondary metabolic diversity of the callus cell lines of traditional Chinese medicinal plant Glycyrrhiza sp. (Glycyrrhiza) by combined chemical analysis and HPLC fingerprint. These callus induction conditions included two Glycyrrhiza species, two types of explants, light and dark conditions, and two combinations of hormones. The evaluation was firstly based on the contents of total flavonoids in the callus by chemical analysis and one way ANOVA. The content of total flavonoids in callus was significantly (P < 0.05) influenced by Glycyrrhiza species, light condition, and the combination of hormones. The callus was further evaluated using diversity factor based on the comparison of HPLC fingerprints of these callus cell lines. Diversity factor varies significantly for calli induced under different conditions, with the highest being at 0.45 under light condition and combination of hormones. These results provide important knowledge for the selection of suitable callus cell lines for the production of pharmacologically important secondary metabolites or bioactive fractions by in vitro culture of Glycyrrhiza sp.
Cell Culture Techniques ; methods ; Cell Line ; Darkness ; Flavonoids ; biosynthesis ; Glycyrrhiza ; cytology ; drug effects ; metabolism ; radiation effects ; Plant Growth Regulators ; pharmacology ; Plant Roots ; metabolism

This paper is aimed to establish the method of fingerprint analysis of chemical constituents by reversed-phase high-performance liquid chromatographic (HPLC) with diode array detector (DAD) for the quality control of the flower buds of Tussilago farfara L. (Flos Farfarae). The method was performed on a Dikma Diamonsil C18 column (250 mm x 4.6 mm ID, 5 gim) with a mixed mobile phase of 0.03% trifluoroacetic acid solution and acetonitrile in a gradient mode. The flow rate was 1.0 mL x min(-1) and the wavelength of measurement was 240 nm. Ten batches of the Flos Farfarae were determined. The HPLC chromatographic fingerprint of chemical constituents was established from the 10 batches of the Flos Farfarae and showed 25 characteristic common peaks, among which 16 peaks were recognized and 18 compounds (adenosine, uridine, gallic acid, 3-O-caffeoylquinic acid, p-hydroxybenzoic acid, trans-caffeic acid, phthalic acid, rutin, hyperoside, 3,5-O-dicaffeoylquinic acid, kaempferol-3-O-beta-D-glucopyranoside, isoferulic acid, ferulic acid, quercetin, 2,2-dimethyl-6-acetyl chromanone, dibutylphthalate, tussilagone, 7beta-(3'-ethylcrotonoyloxy)-1alpha-(2'-methyl-butyryloxy)-3,14-dehydro-E-notonipetranone) were determined by comparison with chromatographic behaviors and UV spectra of the authentic compounds. The 10 batches of samples were classified as 2 clusters by cluster analysis and 6 samples were confirmed to establish the mutual model. The samples' quality was assessed by Similarity Evaluation System for Chromatographic Fingerprint of TCM (2004 B version). The convenient and high specific method can be used to identify and evaluate the quality of the Flos Farfarae.
Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Tussilago ; chemistry

Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Hypothalamus ; metabolism ; Male ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Sprague-Dawley

Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Active Transport, Cell Nucleus ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Human Umbilical Vein Endothelial Cells ; Humans ; NF-kappa B p50 Subunit ; metabolism ; Software

Objective: To test the feasibility of drag-reducing polymers (DRP) for improving coronary microcirculation in a canine model in order to provide the experimental basis for treating myocardial microcirculation dysfunction. Methods: A total of 8 dogs received open-chest surgery and they had intravenous injections, in turn, with adenosine (ADN), DRP 250 mg/L and DRP+ADN. The function y=A × (1-e-βt) was used to calculate the myocardium capillary volume (A value), capillary velocity (β value) and myocardial blood lfow (A ? β value) by myocardial contrast echocardiography. Results: With DRP infusion, the A value in experimental canine was similar to the baseline condition,P>0.05; while theβ value and A ? β value were signiifcantly increased as (0.57 ± 0.10) 1/s vs (0.23 ± 0.03) 1/s,P<0.01 and (11.51 ± 1.96) VI/s vs (5.15 ± 0.86) VI/s,P <0.05 respectively. With combined infusion of DRP+ADN, the β value and A ? β value were similar to the baseline condition, bothP>0.05. Conclusion: DRP improved coronary microcirculation primarily by modulating the β value in experimental canine model, and hopefully, this unique hemodynamics could provide a new approach for treating myocardial microcirculation dysfunction.

Objective:The aim of this study was to detect the P2X7 receptor (P2X7R) protein expression in pancreatic carcinoma and to analyze its association with clinicopathology features of pancreatic carcinoma.And furtherly to explore the effects and underlying mechanisms of P2X7R on invasion and migration of PANC-1 cell line.Methods:P2X7R expression was determined by immunohistochemistry in specimens of primary cancer and adjacent noncancerous tissues respectively,and analyzed the relationship between P2X7R expression and clinicopathology features of pancreatic carcinoma.The transwell assay and wound healing assay were used for investigating cell invasion and migration ability of PANC-1,and western blotting was performed to measure the expresions of MMP2 and MMP9.Results:P2X7R protein was highly expressed in both PANC-1 cell line and tumor tissue,and associated positively with the histological differentiation and lymph node staging.The active P2X7R could increase the cell migration and invasion ability of PANC-1 cell line through up-regulated MMP2 and MMP9.Conclusions:The overexpression of P2X7R plays crucial roles in the migration and invasion of pancreatic carcinoma,and may represent a novel molecular target in pancreatic carcinoma therapy.

Objective To investigate the inducing effects of selenium dioxide(SeO2 ) on the apoptosis in human cervical carcino-ma cell line Hela and its influence on the expression of apoptosis-related proteins caspase-3 and P53 .Methods Hela cells were trea-ted with different concentrations of SeO2 for 24 h in vitro ;the morphological changes of Hela cells were observed by the optical mi-croscope;the influence of SeO2 on the cell proliferation and vitality was examined by the MTT assay ;the flow cytometry was em-ployed to detect the cell apoptosis rate ;the expressions of caspase-3 and P53 proteins in Hela cells were determined by the Western blot analysis .Results Under the optical microscopy ,SeO2 generated the obvious influence on the cell growth morphology ,a large number of cells became rounded and shrunken ,and lost the normal form ,while the adherence cell number was evidently decreased and the proliferation was slowed down ;the MTT results showed that SeO2 markedly inhibited the cell proliferation and viability in a dose-dependent manner ,in which ,the cell apoptosis rates induced by the 0 ,1 .875 ,3 .750 ,7 .500 ,15 .000 and 30 .000 μmol/L con-centrations of SeO2 were 3 .12% ,30 .56% ,33 .42% ,37 .50% ,45 .43% and 69 .38% respectively ,which revealing the obviously in-creasing trend;the Western blot assay revealed that SeO2 could up-regulate the caspase-3 and P53 levels ,and reached the peak value at the concentration of 7 .500μmol/L .Conclusion SeO2 could induce the cervical cancer cell apoptosis possibly by up-regulating the expressions of caspase-3 and p53 in Hela cells .