Adult ; Cranial Nerve Neoplasms ; diagnosis ; Facial Nerve ; pathology ; Female ; Glomus Jugulare Tumor ; diagnosis ; Humans ; Neurilemmoma ; diagnosis

Objective To study the expression of mdm2 genes in nephroblastoma in children and relationship between mdm2 gene and clinical pathological parameters.Methods The protein expressions of mdm2 in 24 cases of nephroblastoma were detected with S-P immunohistochemical method.The relationships between mdm2 expression and clinicopathological parameters were analyzed.Results The positive rates of mdm2 in 24 cases of nephroblastoma were correlated with lymph node metastasis,clinical stage and tumor differentiation.There was a positive relationship between mdm2 protein expression and clinicopathological parameters such as lymph node metastasis,clinical stage and degree of differentiation(P

Adolescent ; Adult ; Humans ; Male ; Rhinitis ; microbiology ; Sinusitis ; microbiology ; Tuberculosis

Objective　To investigate the effect of cardiopulmonary bypass (CPB) on vascular endothelial cell injury and plasma endothelin-1 and nitric oxide equilibrium in patients undergoing cardiovascular operation with CPB. Methods　A total of 20 patients with congenital heart disease (Group Ⅰ) and 20 with valvular problem (group Ⅱ) were operated on under CPB respectively. Blood samples were collected from central vein before skin incision, before CPB, 30 min after CPB, at the end of CPB, and end of operation, the first morning and third morning after operation. The levels of plasma thrombomodulin(TM), endothelin-1(ET-1) and nitric oxide(NO) were measured. Results　The plasma TM level was significantly elevated during CPB (P<0.01, P<0.05) and 1 d after operation, reached its peak as (4.88±1.12) ng/ml in Group Ⅰand (8.34±1.84) ng/ml in group Ⅱ at the end of surgery and came back to the level as before operation. The plasma level of ET-1 was also increased significantly after CPB and reached peak as (129.04±22.29) in Group Ⅰ and (156.62±29.66) in Group Ⅱ at the end of operation. And the level was still higher than before operation in 2 groups 3 d after operation. No change was found on the level of NO in 2 groups. Conclusion　CPB may cause extensive acute endothelial cells damage for about 24-48 h and recovered about 72 h and it may also cause an imbalance of ET-1 and NO.

AIMTo identify the main metabolites of oxymatrine (OMT) in rats.

METHODSTo optimize the conditions of LC/ESI-ITMS' chromatograms and spectra by oxymatrine and matrine (MT), and summarize their ionization and cleavage rules in ESIMS, then serving as the basis for the metabolite analyses of oxymatrine in rats. To collect the 0-24 h urine samples of the rats after ip 40 mg x kg(-1) oxymatrine, the samples were enriched and purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC/ESI-ITMS. The structures of OMT metabolites were identified according to their retention times and ESI-ITMSn rules.

RESULTSSix phase I metabolites and the parent drug OMT were found in the rat urine, and the main metabolite was MT. No phase II metabolites were found.

CONCLUSIONThe developed LC/ESI-ITMSn methods to identify the metabolites of oxymatrine in rats is not only simple and rapid but also sensitive and specific. This technology is one of the most efficient methods for the analysis of drug metabolites.

Alkaloids ; isolation & purification ; pharmacokinetics ; urine ; Animals ; Chromatography, Liquid ; methods ; Plants, Medicinal ; chemistry ; Quinolizines ; isolation & purification ; pharmacokinetics ; urine ; Rats ; Rats, Wistar ; Sophora ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

AIMTo establish a LC-MS(n) method for the identification of anisodamine and its metabolites in rat feces.

METHODSFeces samples were collected after single administration of 25 mg x kg(-1) anisodamine to rats, and dipped in water for 1 h. Samples were then extracted by ethyl acetate. The pretreated samples were separated on a reversed-phase C18 column using a mobile phase of methanol / 0.01% triethylamine (adjusted to pH 3.5 with formic acid) (60 : 40, v/v) and detected by LC-MS". Identification of the metabolites and elucidation of their structures were performed by comparing their changes in molecular masses (deltaM), retention-times and full scan MS(n) spectra with those of the parent drug and blank feces.

RESULTSThe parent drug and its seven metabolites (6beta-hydroxytropine, nor-6beta-hydroxytropine, aponoranisodamine, apoanisodamine, noranisodamine and hydroxyanisodamine, tropic acid) were found in rat feces.

CONCLUSIONThis method is sensitive, rapid, simple, effective, and suitable for the rapid identification of drug and its metabolites in biologic samples.

Animals ; Feces ; chemistry ; Rats ; Rats, Wistar ; Solanaceous Alkaloids ; analysis ; metabolism ; Tandem Mass Spectrometry ; methods

OBJECTIVESTo investigate the cure effect of tumor antigen specific CTL on a model of human hepatocellular carcinoma in nude mice LCI-D20.

METHODSDendritic cells (DCs) were induced from peripheral blood mononuclear cells of healthy people in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4) and were pulsed with tumor antigen from hepatocellular carcinoma cell line MHCC97H. Then tumor antigen specific cytotoxic T lymphocytes (CTLs) were induced. By intraperitoneal injection of tumour antigen specific CTLs into the LCI-D20, the preventive and therapeutic effects of these CTLs to HCC in the LCI-D20 model were assessed. Cytokine-induced killer (CIK) cells and phosphate buffer solution were used as controls at the same time.

RESULTSThe weights of tumors in the tumor antigen specific CTL group, in the CIK cell group and in the blank group were (1.11+/-0.63), (1.12+/-0.36) and (2.68+/-0.53) grams respectively (t = 5.18, t = 6.06, P < 0.01). The amount of blood alpha fetal protein in the tumor antigen specific CTL and CIK groups were (52.1+/-9.7) microg/L and (48.6+/-5.2) microg/L, and was (82.2+/-7.2) microg/L in the blank group (t = 17.26, t = 22.07, P < 0.01 respectively). The metastasis rates in livers were 16.7%, 16.7% and 58.3% in the tumor antigen specific CTL, CIK cell and blank control groups respectively (chi2= 4.44, P < 0.01). The survival time of the mice in the tumor antigen specific CTL group was (79.0+/-5.02) days, (73.3+/-7.0) days in the CIK group, and (52.3+/-5.2) days in the blank group (t = 14.56, t = 17.54, P < 0.01).

CONCLUSIONTumor antigen specific CTLs may prevent metastasis in the LCI-D20 model and prolong the survival time.

Animals ; Antigens, Neoplasm ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Interleukin-4 ; pharmacology ; Liver Neoplasms ; immunology ; pathology ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Recombinant Proteins ; T-Lymphocytes, Cytotoxic ; immunology

AIMTo identify the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine.

METHODSUrine samples from 0 - 24 h were collected after single ig dose of 500 mg x kg(-1) daidzein to each of six rats. The urine samples were purified by SPE column (SPE C18) and analyzed with liquid chromatographic-tandem electrospray ionization ion trap mass spectrometry (LC-ESI/MS(n)) for potential metabolites.

RESULTSSeveral new hydroxylate metabolites and its sulfate conjugates were found and identified in rat urine.

CONCLUSIONLC-ESI/MS(n) is proved to be a simple, rapid, sensitive and specific technique for identification of the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine.

Animals ; Chromatography, Liquid ; methods ; Hydroxylation ; Isoflavones ; chemistry ; metabolism ; urine ; Male ; Molecular Structure ; Phytoestrogens ; chemistry ; metabolism ; urine ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Seeds ; chemistry ; Soybeans ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Sulfates ; metabolism ; Tandem Mass Spectrometry ; methods

AIMTo identify the main metabolites of jatrorrhizine in rat urine.

METHODSThe rat urine samples were collected 0 - 72 h after ig 12 mg x kg(-1) jatrorrhizine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by combining liquid chromatography and tandem electrospray ionization ion trap mass spectrometry (LC-ESI/ITMS(n)). Identification and structural elucidation of the metabolites were performed by comparing the changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug.

RESULTSAt least seven phase I metabolites (such as de-methyl, de-hydrogen and hydroxyl metabolites) and eleven phase II metabolites (such as glucuronide conjugates and methyl-conjugates) were identified in rat urine.

CONCLUSIONThe developed LC-ESI/ITMS(n) method is not only simple and rapid but also sensitive and specific for the identification of metabolites of jatrorrhizine in rat urine.

Animals ; Berberine ; analogs & derivatives ; isolation & purification ; metabolism ; urine ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Spectrometry, Mass, Electrospray Ionization ; methods

AIMTo estabilish a rapid and sensitive LC-ESI-ITMSn method for the identification of ephedrine and its main metabolites in rat urine.

METHODSAfter optimizing the detection condition of LC-ESI-ITMSn chromatography and mass spectrometry by using a standard ephedrine, the ionization and cleavage rules of ephedrine in ESI-MS and ESI-MSn modes were summarized, and then serving as the basis for the metabolite analysis of ephedrine in rat urine. Rat urine samples of 0-48 h were collected after ig 10 mg x kg(-1) ephedrine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC-ESI-ITMSn.

RESULTSThe structures of ephedrine metabolites were elucidated according to the changes of the molecular weights of the metabolites (deltaM) and their cleavage pattern in ESI-ITMSn. As a result, three phase I metabolites and the parent drug ephedrine were identified existing in rat urine, but no phase II metabolites were found.

CONCLUSIONThe LC-ESI-ITMSn method is rapid and highly sensitive and sepecific, it is suitable for the identification of ephedrine and its metabolites in rat urine.

Animals ; Chromatography, High Pressure Liquid ; methods ; Ephedrine ; chemistry ; metabolism ; urine ; Male ; Molecular Weight ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; methods