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1.
Chinese Pharmacological Bulletin ; (12): 735-739, 2018.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-705117

ABSTRACT

Aim To establish human U87-MG glioma model in nude mice brain and to observe the characteristics of the tumor growth. Methods Human U87-MG glioma cells were cultured in vitro. 5 μL of cell suspension containing 3.0 ×1010·L-1, 4.0×1010·L-1and 5.0×1010·L-1respectively was inocula-ted into the right caudate nucleus of 18 male nude mice brain un-der the guidance of stereotaxic apparatus, separately, whereas another 6 nude mice as the control group, were inoculated into the same volume of Hanks solution. The moving and survival state of rats with gliomas were observed. The examinations of the tumors formation, volumes, metastasis and histopathology were performed and the obtained brain samples were stained with HE and immunohistochemistry. Results All the tested rats of dif-ferent inoculation doses developed brain tumors without extracra-nial metastasis. The mean survival time of three groups was (46.50 ± 3.27) d,(38.50 ± 3.28) d and (30.67 ± 3.51) d,respectively. The tumors showed the similar morphological fea-tures and immunophenotype to human glioma. There was positive expression of GFAP and S-100 in the tumors. Conclusions The orthotopic implantation model of human U87-MG glioma, by in-oculating quantitative U87-MG cells stereotaxically into the brains of the nude mice, is successfully established with 100 yield of intracranial tumor and no extracranial growth extension. It resembles the histopathological and morphological features of human glioma,which can be used as a reliable animal model for the study of the tumorigenesis, pathogenesis, biological charac-teristics and therapy of glioma.

2.
Pharmazie ; 70(4): 219-24, 2015 Apr.
Article in English | MEDLINE | ID: mdl-26012250

ABSTRACT

A highly sensitive and rapid liquid chromatography-tandem mass spectrometry method was developed for the determination of clinofibrate in human urine. The analyte and IS were extracted through a simple protein precipitation by the mixture of acetonitrile and 1 mol/L hydrochloric acid (95:5, v/v) and separated on an Inspire C18 (150 mm x 4.6 mm I.D., 5 µm particle size) column using isocratic elution with methanol and water containing 0.1% formic acid and 10 mM ammonium acetate (90:10, v/v). Mass spectrometric detection was performed in electrospray positive ionization MRM mode. The mass transition was m/z 486.3-->175.0 for clinofibrate and m/z 361.1-->233.1 for IS, respectively. The flow rate was 0.6 mL/min and the column oven temperature was set at 35 °C. The total run time was 6.5 min. Good linear relationships were obtained for all analytes over the concentrations ranging of 0.1002-10.02 µg/mL (r2 = 0.9991) and the limit of quantification was 0.1002 µg/mL. The extraction recovery was larger than 87.4% and intra- and inter-batch precision and accuracy with RSD were all less than 6.5%. The total amount of unchanged clinofibrate excreted in urine was less than 0.34%. This method was successfully applied to the pharmacokinetic study of clinofibrate in human urine.


Subject(s)
Hypolipidemic Agents/urine , Phenoxyacetates/urine , Adult , Chromatography, High Pressure Liquid , Female , Fenofibrate/pharmacokinetics , Fenofibrate/urine , Humans , Hypolipidemic Agents/pharmacokinetics , Male , Phenoxyacetates/pharmacokinetics , Reference Standards , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry
3.
J Ethnopharmacol ; 151(1): 108-13, 2014.
Article in English | MEDLINE | ID: mdl-24095700

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Caesalpinia sappan is a medicinal plant native to China popularly used to treat chronic pelvic inflammation, dysmenorrhea and hysteromyoma. Its main bioactive component is brazilin which had presented antibacterial, anti-inflammatory and anti-platelet aggregation activities. To establish a sensitive, selective, reproducible, and accurate high performance liquid chromatographic (HPLC) method for the quantitative determination of brazilin in plasma, and study the pharmacokinetics of brazilin in rats after intravenous administration of brazilin. MATERIALS AND METHODS: Rats received intravenous injection of 25, 50 and 100mg/kg of brazilin. Concentrations of brazilin in plasma were determined by HPLC method at different time points and all pharmacokinetic parameters were estimated by non-compartmental analysis with WinNonLin 6.2 software. RESULTS: After single intravenous doses of 25, 50 and 100mg/kg brazilin in rats, the main PK parameters were as follows: Cmax were 18.1 ± 4.1, 46.7 ± 8.7 and 82.2 ± 9.6 µg/mL; AUC0-24 were 20.4 ± 4.3, 48.7 ± 6.8 and 90.4 ± 10.3 µgh/mL; and t1/2 were 5.4 ± 1.5, 5.8 ± 0.9 and 6.2 ± 1.2h, respectively. CONCLUSION: It showed that the brazilin was eliminated moderately in rat by intravenous injection route with t1/2 of 6h and showed a dose-dependence profile of Cmax and AUC0-24 at the doses of 25~100mg/kg of brazilin for injection in rats.


Subject(s)
Benzopyrans/chemistry , Benzopyrans/pharmacokinetics , Chromatography, High Pressure Liquid/methods , Animals , Area Under Curve , Benzopyrans/blood , Dose-Response Relationship, Drug , Half-Life , Male , Molecular Structure , Rats , Rats, Sprague-Dawley , Sensitivity and Specificity , Tinidazole/chemistry , Tinidazole/pharmacokinetics
4.
J Pharm Biomed Anal ; 76: 152-6, 2013 Mar 25.
Article in English | MEDLINE | ID: mdl-23319135

ABSTRACT

A convenient and rapid HPLC method was developed for the determination of clinofibrate in human plasma using simple protein precipitation with the mixture of acetonitrile and 1M hydrochloric acid (95:5, v/v) followed by separation using an Inspire C(18) column with isocratic elution. The detection wavelength was 232nm and the flow rate was 1.0ml/min. The mobile phase consisted of acetonitrile and water containing 0.4% ortho-phosphoric acid (73:27, v/v). Linear calibration curve was obtained over the concentrations ranging from 0.5µg/ml to 32µg/ml (r(2)=0.999) with LLOQ of 0.5µg/ml. The RSD in both the intra-run and inter-run precision study was less than 5.4% and the extraction recoveries were above 90.7%. The HPLC method is reproducible and suitable for the quantification of clinofibrate in plasma. This method was successfully applied to the pharmacokinetic studies of clinofibrate in healthy volunteers. The elimination half-lives (t(1/2)) were (20.47±3.44), (18.19±2.62) and (21.51±4.78)h after single oral administration of 200, 400 and 600mg clinofibrate, respectively. The results of WinNonlin software showed that the area under the plasma concentration versus time curve from time 0 to 72h (AUC(0-72)) and peak plasma concentration (C(max)) were linearly related to dose (P>0.05).


Subject(s)
Chromatography, High Pressure Liquid/methods , Hypolipidemic Agents/pharmacokinetics , Phenoxyacetates/pharmacokinetics , Administration, Oral , Adult , Area Under Curve , Calibration , Dose-Response Relationship, Drug , Female , Half-Life , Humans , Male , Reproducibility of Results , Software , Time Factors
5.
Acta Pharmaceutica Sinica ; (12): 771-777, 2009.
Article in English | WPRIM (Western Pacific) | ID: wpr-344107

ABSTRACT

A sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was established for the determination of eplerenone (EP) in human plasma. The plasma samples of EP were extracted with ethyl acetate and separated by HPLC on a reversed phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate water solution-methanol (30 : 70, v/v). EP was determined with electrospray ionization-mass spectrometry (ESI-MS) in the selected ion monitoring (SIM) mode. The calibration curves were linear over the range of 2-4 000 ng x mL(-1) for EP. The lower limit of quantification was 2 ng x mL(-1). The method has been successfully applied in the pharmacokinetic study of the EP tablets. The main pharmacokinetic parameters of EP after oral administration of 25 mg, 50 mg, 100 mg were as follows, t1/2: (4.9 +/- 2.1), (4.7 +/- 1.5), (5.9 +/- 1.2) h; AUC(0-infinity): (4 402 +/- 1 735), (8 150 +/- 2 509), (13 783 +/- 4 102) microg x h x L(-1); and MRT: (6.2 +/- 2.1), (6.6 +/- 1.3), and (7.2 +/- 1.6) h. Parameters of EP after oral administration of multiple doses of 50 mg were as follows, t1/2: (6.1 +/- 1.7) h; AUC(ss): (10 071 +/- 4220) microg x h x L(-1); MRT: (8.1 +/- 2.3) h; and DF: (3.2 +/- 1.0).


Subject(s)
Humans , Chromatography, High Pressure Liquid , Methods , Spectrometry, Mass, Electrospray Ionization , Methods , Spironolactone , Blood , Pharmacokinetics
6.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-297069

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the perioperation medication on the first patient who was operated facial allotransplantation, including immunosuppressive drug and adjunctive drug, so that to search a effective medication schedule to the patient operated facial allotransplantation.</p><p><b>METHODS</b>FK506, MMF, Prednisone and Zenapax was performed as immunosuppressive regiment in perioperative treatment; meanwhile, anti-infectives was administered to take precautions against all sorts of infections, such as bacterium, virus and fungus. Furthermore, all kinds of adjunctive drug, Losec, glucurolactone and so on, was administered to protect those function of stomach, liver, kidney and so on. Clinical observations were made on the signs and symptoms of graft survival or rejection, as well as immunological indexes were tested in laboratory. Biopsies of graft were also made at 30 d after operation. Side effect and complication of drug was monitored, in case the body suffered harm.</p><p><b>RESULTS</b>Facial allograft was survived, and the temperature and color of skin were normal. Swelling of tissue was gradually subsidise after 4 days, and recovered in a half month. The count and ratio between Th and Ts were normal, skin Biopsies of every time had no found of hyperacute or acute rejection, and side effect and complication of drug had no monitored.</p><p><b>CONCLUSIONS</b>The regiment of perioperation medication was successfully performed.</p>


Subject(s)
Adult , Humans , Male , Face , General Surgery , Immunosuppressive Agents , Therapeutic Uses , Tissue Transplantation , Methods , Transplantation, Homologous
7.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-350971

ABSTRACT

<p><b>OBJECTIVE</b>To study the bioactive constituents from tubers of Bolbostemma paniculatum.</p><p><b>METHOD</b>Compounds were isolated by extraction and partition as well as several-chromatographic techniques guided with Pyricularia oryzae bioassay method. Their structures were determined on the basis of spectral analysis and chemical evidence.</p><p><b>RESULT</b>Bisdesmoside (I) was isolated as active compound causing morphological abnormality of Pyncularia oryzae mycelia and elucidated as 3-0-alpha-L-arabinopyranosyl (1-->2)-beta-D-glucopyranosyl-bayogenin-28-O-beta-D-xylopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside.</p><p><b>CONCLUSION</b>I is a new natural product and exhibited significant cytotoxicity against cancer cell lines K-562 and BEL-7402, but no hemolytic activity to rabbit erythrocytes.</p>


Subject(s)
Animals , Humans , Rabbits , Biological Assay , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Cell Proliferation , Cucurbitaceae , Chemistry , Erythrocytes , Hemolysis , K562 Cells , Liver Neoplasms , Pathology , Molecular Conformation , Molecular Structure , Plant Tubers , Chemistry , Plants, Medicinal , Chemistry , Saponins , Chemistry , Pharmacology
8.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-232846

ABSTRACT

<p><b>OBJECTIVE</b>To develop a new high-performance liquid chromatographic (HLPC) method for determination of propofol in human serum.</p><p><b>METHODS</b>Human serum samples were precipitated with 20% perchloric acid and centrifuged to obtain 50 microl of the supernatant for analysis by HPLC coupled with fluorescence detection. The analysis was performed with a C(18) reversed-phase column using a acetonitrile-water (90:10) phase delivered at 1.0 ml/min, with the excitation wavelength of 276 nm and emission wavelength of 310 nm.</p><p><b>RESULTS</b>The calibration curves were linear (r=0.997 5) within the concentration range of 0.05-10 microg/ml, the limit of propofol quantification was 50 ng/ml and the intra- and inter-day precisions were between -/+15%.</p><p><b>CONCLUSIONS</b>The method is accurate, sensitive and simple for propofol determination in clinical anesthesia.</p>


Subject(s)
Humans , Anesthetics, Intravenous , Blood , Chemistry , Chromatography, High Pressure Liquid , Methods , Fluorescence , Propofol , Blood , Chemistry , Reproducibility of Results , Spectrometry, Fluorescence , Methods
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