Objective To determine the content of cholesterol in Gekko Gecko.Methods High performance liquid chromatography was performed on Hypersil ODS2 (4.6 mm?150 mm,5 ?m).The chromatorgraphic conditions were as follows:methanol as mobile phase,flow rate being 1.0 mL/min and detecting wavelength at 208 nm.The column temperature was 40 ℃.Results A good linearity of cholesterol was shown in range of 0.885～14.000 ?g (r=0.999 5),the average recovery was 99.65%,RSD=3.45% (n=6).Conclusions This method is simple,rapid and accurate,and may provide a reliable way for the quantification of cholesterol in Gekko Gecko.

Objective To study the expressions and significances of matrix metalloproteinases-3 (MMP-3) and tissue-inhibitor of matrix metalloproteinase-3 (TIMP-3) in transitional cell carcinoma of bladder.Methods Immunohistochemistry was used to detect the expressions of MMP-3 and TIMP-3 in 48 cases of transitional cell carcinoma of bladder and 20 normal bladder tissues.Results MMP-3 and TIMP-3 in normal bladder tissues did not express.The more grading and staging,the more MMP-3 expression,but no statistical significance(P ＞0.05),by contrast,TIMP-3 expression decreased and has statistically significant in the tumor grading (P ＜ 0.05).Conclusions High expression of MMP-3 and low expression of TIMP-3 is related to the malignancy of transitional cell carcinoma of bladder and may play an important role in the development of transitional cell carcinoma of bladder,as an important reference to judge the degree of malignancy of transitional cell carcinoma of bladder and prognostic indicator.

BACKGROUND: Dendritic cells, the most potent antigen presenting cells known at present, have been extensively used in the immunotherapy as adjuvant. OBJECTIVE: The present study was to construct Der p 2 eukaryotic expression vector and validate its expression in the mouse bone marrow-derived dendritic cells. DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Respiratory Disease,Xinqiao Hospital, Third Military Medical University of Chinese PLA between May and December 2005. MATERIALS: C57BL/6 mice were included. Plambd-Der p 2 was the product of Heska Company, USA.pCI-neo plasmid was provided by the Institute of Respiratory Disease, Xinqiao Hospital, Third Military Medical University of Chinese PLA.METHODS: Mouse bone marrow-derived dendritic cells were in vitro isolated and cultured.Complete Der p 2 cDNA was spliced from prokaryotic expression vector plambd-Der p 2, and then cloned into eukaryotic expression vector pCI-neo (pCI-neo-Der p 2).The positive recombinants pCl-neo-Der p 2 transfected into dendritic cells.Non-transfected and blank vector pCI-neo-transfected dendritic cells were used as controls. MAIN OUTCOME MEASURES: ①Identification of pCI-neo-Der p 2 recombinant plasmid.②Detection of Der p 2 mRNA and protein expression by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot techniques. RESULTS: Sequencing results showed Der p 2 cDNA in pCI-neo-Der p 2 was in coincidence with the sequence registrated in Gene Bank.RT-PCR and Western Blot results showed that expression of Der p 2 mRNA and protein could be detectable in the pCI-neo-Der p 2-transfected dendritic cells. CONCLUSION: The Der p 2 cDNA was successfully constructed into the eukaryotic expression vector, and Der p 2 gene and protein could be expressed efficiently in dendritic cells.

ction of immune tolerance of T cells to allergens.

Objective:To observe the co-antitumor effect and hepatic protection of melatonin on unresectable advanced primary liver cancer treated by transcatheter arterial chemoembolization (TACE). Methods:One hundred and forty patients with unresectable advanced primary liver cancer were divided into 2 groups at random, seventy patients received TACE alone while the other 70 patients were treated by TACE+Mel (20 mg/d at 8:00 P.M orally, 7 d before each TACE and lasted for 21 d). Results: The partly resolution rate of TACE and TACE+Mel were 12.86% and 22.86% respectively( P

Objective To investigate the relationship between the levels of fractional exhaled NO and airway inflammation cells in cough variant asthma(CVA)and the effect of inhaled corticoids .Methods CVA patients were treated with Budesonide and Formoterol Fumarate Powder for Inhalation for 6 months .Before and after treatment ,the provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second(PC20 FEV1 )was tested ,and the levels of fractional exhaled No ,and eo‐sinophil in induced sputum were examined .Results The levels of FeNO ,the percentage of eosinophil in induced sputum were sig‐nificantly higher than control healthy subjects ,and all decreased significantly after treated with ICS/LABA ,together with elevated PC20 FEV1 .The levels of FeNO were significantly correlated with the percentage of eosinophil in induced sputum ,and also negative‐ly correlated to PC20 FEV1 .Conclusion The levels of FeNO in CVA patients were significantly higher than control healthy sub‐jects ,and correlated with the inflammation of airway ,so FeNO may be used as a biomarker to determine the control level of airway inflammation in CVA patients .

AIM To investigate the effect of cisplatin on apoptosis in Scaber cell. METHODS The apoptotic cells were detected by TUNEL,HE,eletronic micrpscopy. RESULTS Treatment of Scaber cells with CDDP resulted in characteristics typical of apoptosis. CDDP induced apoptosis of Scaber cells in time and concentration dependent manner. To further investigate the mechanism of apoptosis induced by CDDP, the expressions and activity of apoptosis associated proteins such as bcl 2, bax and caspase 3 were examined using S P method.The results showed: CDDP caused time and concentration dependent decreases in bcl 2 and increased in bax proteins.CDDP bcl 2 and its translocation to perinuclei and nuclei. The expression of caspase 3 in Scaber cell were determined during apoptosis induced by CDDP. CONCLUSION Our investigetion showed that the apoptosis induced by CDDP is related to the increase of bax protein, and the decrease of bcl 2 protein. and its translocation to perinuclei and nuclei.

Objective To improve the methods for in vitro generating dendritic cells (DCs) from mouse bone marrow and to identify it with morphological, phenotype determination. Methods Cells isolated from mice bone marrow were cultured in GM-CSF (20 ng/ml), differentiating into dendritic cells. Morphological changes were observed by optical phase contrast microscopy, and surface molecules including CD11 C, CD80, MHCⅡ were detected by FACS. Results A large number of typical DCs were observed after culturing for 10 d. FACS analysis showed that the amplified DCs could express CD11 C, CD80, MHCⅡ. Conclusion A large quantity of highly pure BM-DC can be obtained by this method.

Objective: To explore the regulative role of extracellular regulated protein kinase-5 (ERK5)/Bcl-2 interacting mediator of cell death (Bim) pathway in hypothermal stimulation induced neonatal rat’s cardiomyocytes (CMs) damage and apoptosis.
Methods: CMs were cultured for hypothermal stimulation and the speciifc siRNA was used to down-regulate the ERK5 or Bim in CMs. The cell apoptosis was detected by lfow cytometry, protein expression was examined by Western blot analysis, the intracellular Ca2+, reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were evaluated by lfuorescent labeling and lfow cytometry.
Results: In hypothermal stimulated CMs, ERK5 siRNA could promote Bim protein expression, but Bim siRNA could not inlfuence ERK5, while attenuated p-ERK5 expression. ERK5 siRNA induced higher apoptosis rate, while Bim siRNA could decrease such effect. ERK5 siRNA increased the intracellular Ca2+overloading, ROS activation andΔΨm damage, while Bim siRNA played the role to against those effects in hypothermal stimulated CMs.
Conclusion: Our study revealed that ERK5/Bim pathway played the important regulative roll in hypothermal stimulation induced neonatal rat’s CMs damage and apoptosis.

(1)H-NMR and (13)C-NMR assignments of 12-oleanene-3,11-dione (compound 1) were completely described for the first time through conventional 1D NMR and 2D shift-correlated NMR experiments using (1)H-(1)HCOSY, HMQC, HMBC techniques. Based on its NMR data, the assignments of 28-hydroxyolean-12-ene-3,11-dione (compound 2) were partially revised.
Euonymus ; metabolism ; Magnetic Resonance Spectroscopy ; Molecular Conformation ; Oleanolic Acid ; analogs & derivatives ; analysis ; chemistry ; Triterpenes ; analysis ; chemistry