In this study, the flow injection (FI) technology combined with the unique step wise multiple ions monitoring scanning (step-wise MIM) mode of Qtrap-MS was first established for the identification and discrimination of eight Murraya species. It only takes 5 min for each sample to detect approximate 600 compounds. The characteristic MS chromatograms of eight Murraya plants were analyzed by Analyst and SIMCA-P. The results of PCA showed that sect. Murraya and sect. Bergera were clearly divided into two categories, suggesting that there is difference in the chemical compositions between these two groups. Further detail analysis of the MS data could realize the preliminary structure elucidation of the component types contained in different plants. The main components in M. exotica and M. alata are coumarins, and polymethoxyflavones are rich in M. paniculata, while carbazole alkaloids are the major ones in sect. Bergera plants. The results are consistent with our previous comprehensive analysis of the chemical components of Murraya species. In conclusion, our research confirmed that FI-Qtrap-MS technology can be used for rapid identification and differentiation of similar plant species, providing reference for chemical taxonomy and a new method for the quality evaluation of medicinal materials.

Fifty-six patients with Graves' ophthalmopathy(GO)were treated with antithyroid drug and oral prednisone for three months,TSH receptor antibody(TRAb)level was reduced,GO activity and severity of some patients were ameliorated but still positively associated with TRAb.It suggests that TRAb not only triggers off GO but also plays a possible role in the maintenance of the autoimmune process in GO.

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Medicine, Tibetan Traditional ; Phyllanthus emblica ; chemistry ; classification ; Quality Control ; Tannins ; analysis ; Tibet

AIMTo observe the effect of stretching left ventricles in the end of action potential on rabbit cardiac activity, and to investigate its possible mechanisms.

METHODSStretch (120 mmHg, 50 ms) was applied in the end of action potential by the pressure-clamp technique to observe if there would be any changes in rabbit cardiac activity and streptomycin (500 micromol/L) was used to identify the mechanism.

RESULTSStretch in the end of action potential caused arrhythmia (P < 0.05) and streptomycin blocked the above effect (P < 0.05).

CONCLUSIONStreptomycin could block the effect of stretching left ventricles in the end of action potential on rabbit cardiac activity, which indicates that stretch-activated ion channels involve it.

Action Potentials ; physiology ; Animals ; Arrhythmias, Cardiac ; etiology ; physiopathology ; Female ; Heart Ventricles ; physiopathology ; In Vitro Techniques ; Ion Channels ; physiology ; Male ; Mechanoreceptors ; drug effects ; Proprioception ; Rabbits ; Streptomycin ; pharmacology

AIM:To investigate the effects of evodiamine on the growth and apoptosis of human hepatocellular carcinoma Huh7 cells,and to illustrate the molecular mechanism that evodiamine enhances antitumor activity of tumors nec -rosis factor-related apoptosis-inducing ligand(TRAIL)in Huh7 cells.METHODS: The cell viability was measured by MTT assay.The cell cycle distribution was analyzed by flow cytometry.The apoptosis rate was determined by TUNEL stai-ning.The protein levels of cell cycle-and apoptosis-related proteins were detected by Western blot analysis.RESULTS:Treatment of Huh7 cells with evodiamine reduced the cell viability(P<0.05).Evodiamine induced cell cycle arrest in G2/M phase by upregulation of p27,cyclin B1, cell division cycle protein 2(Cdc2)and p-Cdc2.Evodiamine triggered apoptosis accompanied by cleavage of caspase-3 and poly(ADP-ribose)polymerase(PARP).Combination of evodiamine with TRAIL significantly reduced the cell viability and increased cleavage of caspase -3 and PARP as compared with the use of each agent alone.Moreover,evodiamine increased the expression of death receptor 5(DR5)in the Huh7 cells.CON-CLUSION:Evodiamine inhibits the cell growth by reducing the cell viability and inducing cell cycle arrest.Evodiamine also triggers cell apoptosis and enhances the sensitivity of Huh 7 cells to TRAIL by upregulating the expression of DR5.

OBJECTIVETo investigate the expression of Slingshot-1L (SSH-1L) in peripheral blood eosinophils of asthmatic patients.

METHODSPeripheral vein blood sample of 30 ml was collected from 15 healthy volunteers, 15 asthmatic patients with glucocorticoid treatment and 15 asthmatic patients without the treatment. The eosinophils were isolated, purified and counted for each sample, and SSH-1L/beta-actin gene fragments were amplified simultaneously by RT-PCR for the total RNA. SSH-1L protein was detected by Western blotting from the total protein of the peripheral eosinophils. The expressions of SSH-1L at both mRNA and protein levels are compared between different groups.

RESULTSSSH-1L/beta-actin ratio significantly increased in untreated patients with asthma attacks in comparison with healthy volunteers (P<0.05), which did not occur in patients treated with glucocorticoids (P>0.05). The optical density of SSH-1L protein significantly increased in untreated asthmatic patients (P<0.05), but not in patients treated with glucocorticoids (P>0.05), as compared with the healthy volunteers.

CONCLUSIONSignificantly increased SSH-1L expression in peripheral eosinophils may play an important role in the activation and migration of eosinophils.

Actin Depolymerizing Factors ; metabolism ; Actins ; metabolism ; Adult ; Asthma ; blood ; Blotting, Western ; Eosinophils ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Phosphoprotein Phosphatases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

The study was purposed to investigate the effect of arsenic trioxide (As(2)O(3))- induced p16 gene demethylation by a sensitive and specific PCR-based method (nested-methylation specific PCR, n-MSP) and DNA sequencing for rapid analysis of the promoter demethylation status, and to explore the possible mechanism of the p16 gene demethylation in human multiple myeloma U266 cells induced by As(2)O(3). The methylation status of the p16 gene in U266 cell line before and after treatment with As(2)O(3) was detected by the nested-methylation specific PCR and DNA sequencing, the mRNA of p16, DNA methyltransferase (DNMT 1, DNMT3A and 3B) gene were determined by RT-PCR, and the induced growth inhibition of U266 cell was assayed by growth curve, MTT and CFU; the DNA content of U266 cells was analyzed by flow cytometry after being exposed to As(2)O(3). The results showed that (1) all cytosines in CpG dinucleotides in untreated U266 cell not were changed, while all cytosines in treated U266 cells with As(2)O(3) had been converted to thymidine. (2) p16 gene was not expressed in U266 cell line after methylation. As compared with the beta-actin, the expression of U266 cell p16 gene mRNA was increased to (0.22 +/- 0.10), (0.59 +/- 0.11), (0.68 +/- 0.09) after exposed to 0.5 micromol/L, 1.0 micromol/L and 2.0 micromol/L As(2)O(3) for 72 hours respectively. (3) As(2)O(3) could significantly down-regulate DNA methyltransferase 1 (DNMT 1), DNMT3A and DNMT3B gene at mRNA level in a dose-dependent manner. (4) U266 cells line grew slowly and arrested at G(0) - G(1) phase after treatment with three different concentrations of As(2)O(3). It is concluded that As(2)O(3) can activate and up-regulate the expression of p16 gene which inhibits the proliferation of U266 cell through inducing the G(0) - G(1) arrest by demethylation or/and by inhibiting DNMT 1, DNMT3A and 3B gene.
Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Base Sequence ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; biosynthesis ; genetics ; DNA Methylation ; drug effects ; Genes, p16 ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Oxides ; pharmacology ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects

OBJECTIVETo explore the clinical significance of serum free light chain (sFLC) levels in nonsecretory multiple myeloma (NSMM).

METHODSNine NSMM patients were hospitalized in our department from Feb 2002 to Sep 2006 and no M-components was found in their serum and urine by immunofixation electrophoresis (IFE). sFLC was assayed by immuno-nephelometry. The clonality of sFLC was estimated by serum kappa:lambda sFLC ratio. Meanwhile, serum immunoglobulin, total kappa and lambda light chain level were also determined in these patients.

RESULTSIncreased serum concentrations of either kappa or lambda sFLC (and abnormal kappa/lambda ratios) were detected in 6 of 9 patients with NSMM although their serum immunoglobulin levels were not elevated and total kappa:lambda light chain ratios (1.32 - 2.20) were in the reference range. All the 9 patients had clonal IgH gene rearrangements.

CONCLUSIONQuantification of sFLC by immuno-nephelometry is more sensitive than that of serum total light chain measurement and is helpful in estimating the clonality of the light chain in patients with NSMM.

Adult ; Female ; Humans ; Immunoglobulin Light Chains ; blood ; Male ; Middle Aged ; Multiple Myeloma ; blood ; Nephelometry and Turbidimetry ; Sensitivity and Specificity

OBJECTIVETo analyze the clinical and laboratory features and risk factors of multiple myeloma (MM) with extramedullary disease (EM) and its extraosseous localizations at diagnosis and during the course of MM.

METHODSThe clinical features, survival rate and prognostic factors were retrospectively analyzed in 40 patients having EM from a total of 418 MM patients hospitalized in Changzheng Hospital from 1993 to 2006.

RESULTSAmong the 40 patients, the first three localizations of EM involved soft tissue, pleura or peritoneum and central nervous system (CNS). Median duration of follow-up was 30 months. The median overall survival (OS) was 28 months. Twenty-five patients (6%) were found to have EM at diagnosis (group A), and their median OS was 16 months and 15 patients (3.6%) developed EM during the course of the disease (group B), and their expected median OS was 72 months. There was a significant difference between group A and B (P = 0.0045) for OS. Compared with those in group A, patients in group B had a higher percentage of plasmacytes (P = 0.022) and plasmablasts (P = 0.029) in bone marrow, and less advanced stage for international staging system (ISS) (P = 0.027). Log-rank univariate analysis showed that higher CRP level, higher serum LDH, Stage II and III for ISS, Hb < 110 g/L at diagnosis were poor prognostic factors. However, multivariate analysis with COX model showed none of them were statistically significant.

CONCLUSIONEM tumors are not a rare manifestation of MM. Soft tissue in the commonest area involved. Higher serum CRP and LDH level, more advanced stage for ISS, anemia and having EM are poor prognostic factors of MM.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; complications ; pathology ; therapy ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Analysis

OBJECTIVEEosinophils play a pivotal role in asthmatic airway inflammation. We previously found a significantly high expression of Slingshot-1L (SSH-1L) in peripheral eosinophils in acute exacerbations of asthma. Objective To investigate the expression and localization patterns of SSH-1L in peripheral blood eosinophils of asthmatic patients and their changes after treatment with inhaled corticosteroids.

METHODSWe recruited 4 outpatients with acute exacerbations of asthma who received no previous corticosteroid treatment and 1 healthy volunteer. From all the subjects 30 ml peripheral venous blood samples were collected before and after a 3-month treatment with inhaled fluticasone. The eosinophils were isolated, purified and counted, and the expressions of SSH-1L in the eosinophils were examined by RT-PCR and Western blotting. The localization of SSH-1L phosphatases in the peripheral eosinophils was detected by immunofluorescence assay in one patient.

RESULTSSSH-1L phosphatases distributed diffusely in the cytoplasm, especially dense near the membrane of the peripheral eosinophils. Glucocorticoids treatment resulted in a significant reduction in both the SSH-1L mRNA expression (0.7403∓0.1124 vs 0.4101∓0.0363, P=0.001) and SSH-1L protein expression (0.3410∓0.1337 vs 0.1543∓0.0551, P=0.039).

CONCLUSIONA high expression of SSH-1L in peripheral eosinophils in acute exacerbations of asthma may play a role in the activation and migration of eosinophils. The efficacy of inhaled corticosteroids in asthma control might be partly attributed to a down-regulated expression of SSH-1L.

Adult ; Aged ; Asthma ; blood ; drug therapy ; Eosinophils ; metabolism ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Middle Aged ; Phosphoprotein Phosphatases ; metabolism