Two-dimensional electrophoresis(2-DE) is an important technique in proteomics research.The 2-DE system for proteome of Monascus ruber was established by comparing and analyzing the infection caused by different kinds of mediums,lysis buffer and the condition of rehydration.By cultivating the Monascus ruber with YES for 6 days,extracting total protein by TCA-acetone,lysis buffer with 8 mol/L urea,2 mol/L thiourea,4 % CHAPS,1 % DTT and 2 % Bio-lyte,an ideal 2-DE map with higher resolution and better legibility was obtained,which laid a foundation for the further studies on proteome of Monascus ruber.

OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Solitary Fibrous Tumors ; metabolism ; pathology

Adolescent ; Adult ; Aged ; Cadherins ; genetics ; metabolism ; physiology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; genetics ; metabolism ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; physiology ; Middle Aged ; Neoplasm Invasiveness ; physiopathology ; Pituitary Neoplasms ; pathology ; physiopathology ; Young Adult

In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.
Carthamus ; chemistry ; Cell Line ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Flowers ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Kidney ; chemistry ; cytology ; drug effects ; Protective Agents ; chemistry ; pharmacology

The paper reported an investigation of the pharmacokinetics of SN-38 (7-ethyl-10-hydroxy-camptothecin) in rats and the tissue distribution in mice after injection of irinotecan hydrochloride nanoparticles (CPT-11) via tail veins. An LC-MS/MS method was established to determine the concentrations of SN-38 in whole blood of rats and in different tissues of mice. The pharmacokinetics and tissue distribution of SN-38 were compared after the intravenous injection of CPT-11 NPs and CPT-11 solution. Compared with irinotecan solution, the elimination half-life of SN-38 was prolonged from 2.17 h to 2.67 h after the intravenous injection of CPT-11 NPs, but its AUC had little change. After the injection of CPT-11 NPs in mice, over time, the concentrations of CPT-11-metabolized SN-38 in CPT-11 NPs were significantly higher in the whole blood, colon and lungs than those in CPT-11 solution, followed by in the spleen and liver, but those in the heart and brain had no change. However, the amount of SN-38 in the kidneys was reduced with time. CPT-11 NPs could prolong SN-38's (one of its metabolites) blood circulation time in rats and significantly increased the concentration of CPT-11-metabolized SN-38 in the whole blood, colon and lungs of mice. CPT-11 NPs made SN-38 efficiently target-bind to the colon and lungs of mice.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacokinetics ; Camptothecin ; analogs & derivatives ; pharmacokinetics ; Chromatography, Liquid ; Colon ; metabolism ; Half-Life ; Injections, Intravenous ; Lung ; metabolism ; Mice ; Nanoparticles ; administration & dosage ; Rats ; Tandem Mass Spectrometry ; Tissue Distribution

Objective To investigate the fluctuation of the iodine deficiency disorders(IDD) through continuous monitoring on it. Methods During 2005-2008, 10 monitoring countis (city,district) of IDD were selected in Jiangxi Province. Five townships were selected in each monitoring county, one school was selected in each townships, and 40 students in 8-10 years old from each school were selected to inspect their thyroid glands and determined the idodine content of salts from their home. Among the 40 students, 20 of which were selected to determine the intelligence quotient(IQ) and 6 of which were selected to determine urine iodine. Thirty pregnant women were also selected to determine the urine iodine in each monitoring county. Twenty students of 5th grade in each school and 5 housewives living nearby the school were selected to carry out the questionnaire survey of health education about IDD. Results Monitoring results showed the average goiter rote of 8-10 years old students was between 2.00% and 4.60% during 2005-2008. The rate of IQ of the students higher than 90 was all above 76% each year. The median of urine iodine of students and pregnant women was higher than 100 μg/L, and the proportion of people whose urine iodine content was lower than 50 μg/L was below 4% each year. The rate of inhabitant taking qualified idodine salt wasn't lower than 95%. The pass rate of the questionnaire survey of health education about IDD was between 11.67% and 43.50% in students and 92.50%-99.60% in housewives. Conclusions The situation of iodine deficiency in Jiangxi Province has been greatly improved, reaching the standard of eliminating IDD.

Objective To explore oropharyngeal swallowing disorders with videofluoroscopic swallowing study (VFSS). Methods 16 patients with dysphagia accepted VFSS with 10 ml of thin barium meal (50% w/v), thick barium meal (270% w/v), biscuit coated with thick barium meal in single swallow. Their swallowing function was observed on the lateral and anterior/posterior planes, including: symmetry of pyriform sinuses, oral transit time, presence of pharyngeal delay, pharyngeal transit time, oral and pharyngeal residue, and presence of aspiration.Results 5 patients demonstrated oral swallowing disorder. 3 patients demonstrated pharyngeal swallowing disorders, that was pharyngeal delay which caused in aspiration after swallowing. 8 patients demonstrated oropharyngeal swallowing disorders, and 3 of them presented aspiration,2 patients were silent aspirators, 1 was aspiration before and 1 after swallowing. The aspiration time could not be judged from the videofluoroscopy in the other one. For 4 patients with aspiration, 3 were severe, with more than 25% of the bolus aspirated, and 1 aspirated less than 5%. Conclusion VFSS can be helpful to plan individual rehabilitation.

The metabolomic analysis of three Cimicifuga species was performed using H-NMR spectroscopy and pattern recognition (PR) techniques. A broad range of metabolites could be detected by 'H-NMR spectroscopy without any chromatographic separation. The analysis using principal component analysis (PCA) and discriminant partial least square (DPLS) of the 1H-NMR spectrum showed a clear discrimination between C. foetida and the other two species. The major metabolites responsible for the discrimination were triterpenoid saponins and saccharides. These results indicated that the combination of 1H-NMR and PR provides a useful tool for chemotaxonomic analysis and authentification of Cimicifuga species, and could used for the quality control of plant materials.
Cimicifuga ; classification ; Discriminant Analysis ; Drugs, Chinese Herbal ; classification ; standards ; Magnetic Resonance Spectroscopy ; methods ; Metabolomics ; methods ; Pattern Recognition, Automated ; Principal Component Analysis ; Protons ; Saponins ; isolation & purification ; Triterpenes ; isolation & purification

Animals ; Astrocytes ; immunology ; metabolism ; Male ; Microglia ; immunology ; metabolism ; Neurons ; metabolism ; Oxidopamine ; metabolism ; Parkinson Disease ; immunology ; Rats ; Rats, Sprague-Dawley