Objective To explore the effective method for in vitro culture of the dendritic cells(DCs) and the specific anti-leukemic cell effect mediated by dendritic cells pulsed with acute myelogenous leukemia antigen. Methods Bone marrow mononucleas cells (BMMNCs) isolated from AML patients were induced to undergo differentiation with 500 ng/ml A23187 and pulsed with AML antigen. After 96 h, DCs and T cells were co-cultured for 5 to 7 days. The cytotoxic activity of cytotoxic T lymphocyte(CTL) to AML were detected with MTT colorimetry. Results BMMNCs isolated from AML patients treated with 100 ng/ml rhGM-CSF in combination with 500 ng/ml A23187 for 96 h exhibited typical morphology of DCs with rapidly increased expression of CD1a and CD83 (P<0.01). Dendritic cells pulsed with acid eluted tumor antigen peptides group displayed the strongest cytotoxic activity of CTL to AML. There was a significant difference between two groups(P<0.01). Conclusion BMMNCs isolated from AML patients can be successfully induced to DCs with rhGM-CSF and A23187 and dendritic cells pulsed with acid eluted tumor antigen peptides group had the strongest cytotoxic activity of CTL to AML.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Octanes ; analysis ; Workplace

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Pesticides ; poisoning ; Young Adult

Aging ; Health ; Humans ; Longevity

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Drug Overdose ; epidemiology ; Female ; Hospitals, General ; statistics & numerical data ; Humans ; Infant ; Male ; Middle Aged ; Poisoning ; epidemiology ; Young Adult

Azabicyclo Compounds ; blood ; Chromatography, Gas ; Humans ; Piperazines ; blood ; Serum ; chemistry

Objective To compare and evaluate the effect of PBL in clinical teaching of Pediatrics.Methods Among students of Grade 2002 in our university,two types of PBL,pre-learning and case-discussion,were used in their clinical learning of Pediatrics. And then,their effects were evaluated and compared with those of traditional learning method.Results More than 60% of the students agreed with PBL methods,and they considered PBL favorable to practice scientific logical thinking of clinical affairs,to increase their capabilities of learning,oral expression,communication and cooperation.The teachers agreed with PBL methods too for the better learning effect resulting from PBL.Conclusion PBL fits the needs of medical learning reformation.To train new type of doctors in century 21st,it is necessary to use kinds of new learning methods,including PBL methods and standardized patient (SP)in clinical teaching.

Objective The purpose of this paper is to screen inborn errors of meta bolism (IEM) by analyzing urinary components, so as to provide laboratory guide for their diagnosis and therapy.Methods Urine samples of patients suspected to have IEM were collec- ted.Urea was de compo sed with urease and n-heptadecanoic acid was added as internal standard.Protein was denatured with ethanol and precipitate was removed by centrifugation,dried b y evaporation, the residue was trimethylsilylly derivatized with BSTFA/TMCS,and then analyzed with GC-MS for quantification of organic acids, amino acids,suga rs, polyols, purines and pyrimidines, simultaneously. This procedure is denom inated as urease pretreatment-gas chromatography-mass spectrometry (UP-GC-MS) internationally.Results Urinary samples of 327 patients from 6 provinces, cities and autonomous regions were analyzed,and 16 kinds of 27 cases of IEM were screened out with a positiv e rate of 8.26%,among which there were 3 cases of hyperphenylalaninemia,3 cases of glyceroluria,3 cases of Leigh syndrome, 2 cases of propionic acidemia, 2 case s of methylmalonic aciduria, 2 cases of von Gierke′s disease, 2 cases of fructo se-1,6-diphosphatase deficiency, 2 cases of fructosuria, 1 cases of multiple car boxylase deficiency, 1 cases of glutaric acidemia typeⅠ, 1 cases of maple sy rup urine disease, 1 cases of hyperglycinemia, 1 cases of 3-aminoisobutyric acid uria,1 cases of adult-onset typeⅡcitrullinemia,1 cases of galactosemia and 1 ca ses of Fanconi′s syndrome.Several IEM patients above had died,but satisfactory therapeutic effects had been achieved in some diseases,in cluding multiple carboxylase deficiency,methylmalonic aciduria and galactosemia. Other patients′ condition remained to be followed up.Conclusion Analysis of urinary components by UP-GC-MS provides a valuable tool for screenin g of IEM and the results will help to provide effective diagnostic and therapeut ic guide for the patients. J Appl Clin Pediatr,2005,20(2):142-144

To study the effect of Gegen Qinlian decoction and its major effective components on five hepatic microsomal CYP450 isozymes in rats. The in vitro hepatic microsomal incubation technique was used to co-culture Gegen Qinlian decoction and its major effective components together with each probe substrate. HPLC-MS/MS was used to establish the analytical method for metabolites of the five isoform probe substrates of CYP450 isozymes, detect the linearity among micoromal protein concentration, incubation time and metabolite formation amount. And HPLC-MS/MS was applied to determine the formation rate (V) of corresponding metabolites (acetaminophen, 4-OH-chlorzoxazone, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone) specific probe substrates of the five isoform probe substrates of CYP450 isozymes (phenacetin, polbutamide, dextromethorphan, chlorzoxazone, testosterone), in order to determine the activity of each isozyme. The result showed good linearity among acetaminophen, 4-OH-tolbutamide, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone, satisfactory precision, stability and average recovery, suggesting the method was feasible. The optimized in vitro microsomal incubation conditions conformed to the requirements in the guideline of drug-drug interaction. Gegen Qinlian decoction showed different degrees of inhibitor effect on 5 CYP450 isoforms (CYP1A2, CYP2C11, CYP2D2, CYP2E1, CYP3A1/2). Its major effective component berberine could inhibit each CYP450 isoform at high concentrations (except for CYP1A2, CYP3A1/2).
Animals ; Chromatography, High Pressure Liquid ; methods ; Cytochrome P-450 Enzyme Inhibitors ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Isoenzymes ; antagonists & inhibitors ; Liver ; enzymology ; Rats ; Tandem Mass Spectrometry ; methods

Adult ; Hemangioma ; Humans ; Male ; Nose Neoplasms ; Paranasal Sinus Neoplasms