RÉSUMÉ
The properties of polyethyleneimine-cholesterol cationic lipopolymer (PEI-Chol) as gene carries and its gene transfer efficiency in vitro with lipid microbubbles were presented in this paper. PEI-Chol lipopolymer was synthesized by linking cholesterol chloroformate to the amino groups of branched poly(ethyleneimine) (PEI) of 1 800. The structure and molecular weight of PEI-Chol were confirmed by IR, 1H NMR and MADI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry), respectively. The average molecular weight of PEI-Chol was approximately 2 000. The gene delivery system of bubble/PEI-Chol/DNA was constructed by mixed PEI-Chol/pDNA (N/P 10:1) complexes with lipid microbubbles (2-8 microm) which were prepared by DPPC, DSPE-PEG2000 and perfluoropropane with the reverse phase evaporation technique. pEGFP-Cl (enhanced green fluorescent protein) was used as report gene to investigate the DNA condensing ability of PEI-Chol lipopolymer by agarose gel electrophoresis. And their cytotoxicity and in vitro transfer efficiency of different complexes were compared with each other in A549 and MCF-7. The results indicated PEI-Chol lipopolymer can condense plasmid DNA when N/P ratio upto 4, PEI-Chol complexes and bubble/PEI-Chol/DNA complexes were nontoxic to A549 and MCF-7 when formulated at the N/P ratio of 10/1 as determined by MTT assay. This bubble/PEI-Chol/DNA delivery system provided good transfer efficiency with other desirable characteristics such as against-precipitation of plasma proteins. In conclusion, bubble/PEI-Chol/DNA complex is a novel non-viral gene delivery system.
Sujet(s)
Femelle , Humains , 1,2-Dipalmitoylphosphatidylcholine , Chimie , Tumeurs du sein , Anatomopathologie , Lignée cellulaire tumorale , Survie cellulaire , Cholestérol , Chimie , Produits de contraste , ADN , Chimie , Génétique , Fluorocarbones , Chimie , Techniques de transfert de gènes , Vecteurs génétiques , Protéines à fluorescence verte , Métabolisme , Lipides , Chimie , Tumeurs du poumon , Anatomopathologie , Microbulles , Phosphatidyléthanolamine , Chimie , Plasmides , Polyéthylène glycols , Chimie , Polyéthylèneimine , Chimie , Transfection , MéthodesRÉSUMÉ
<p><b>BACKGROUND</b>Severe acute respiratory syndrome coronavirus (SARS-CoV) is a newly emerging virus that gives rise to SARS patients with high rates of infectivity and fatality. To study the humoral immune responses to SARS-CoV, the authors evaluated IgG and IgM specific antibodies in patients' sera.</p><p><b>METHODS</b>Two methods, enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent assay (IFA), were used to detect specific serum IgG and IgM against SARS-CoV in 98 SARS patients and 250 controls consisting of patients with pneumonia, health-care professionals and healthy subjects. The serum antibody profiles were investigated at different times over one and a half years in 18 of the SARS patients.</p><p><b>RESULTS</b>The sensitivity and specificity of ELISA for detecting IgG against SARS-CoV were 100.0% and 97.2% and for IgM 89.8% and 97.6% respectively; the figures using IFA for IgG were 100.0% and 100.0% and for IgM 81.8% and 100.0% respectively. During the first seven days of the antibodies trace test, no IgG and IgM were detected, but on day 15, IgG response increased dramatically, reaching a peak on day 60, remaining high up to day 180 and decreasing gradually until day 540. On day 15, IgM was detected, rapidly reached a peak, then declined gradually until day 180 when IgM was undetectable.</p><p><b>CONCLUSION</b>The detection of antibodies against SARS virus is helpful in the clinical diagnosis of SARS.</p>
Sujet(s)
Adulte , Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Anticorps antiviraux , Sang , Test ELISA , Technique d'immunofluorescence indirecte , Immunoglobuline G , Sang , Immunoglobuline M , Sang , Virus du SRAS , Allergie et immunologie , Syndrome respiratoire aigu sévère , Allergie et immunologie , Lymphocytes T cytotoxiques , Allergie et immunologieRÉSUMÉ
<p><b>OBJECTIVE</b>To observe the effects of Epimedium total Flavonoids Phytosomes on preventing and treating bone-loss of the castrate osteoporosis rat model.</p><p><b>METHOD</b>The osteoporosis model was established with 4-month-odl panther's rats, their ovaries on both sides castrated. Dual energy X-ray scanning was used to determine the bone density, and immunity and ELASA were used to assay concentration of estradiol and IL-6 in serum respectively, then determine their effect.</p><p><b>RESULT</b>The BMP and E2 of high dosage group nilestriol group and normal group are higher than those of model group (P < 0.01), while their content of IL-6 is apparently lower than that of model group(P < 0.01).</p><p><b>CONCLUSION</b>The osteoporosis model was established successfully and the using of EFP can improve the bone density, enhance E2 level and decrease the IL-6 concentration in serum.</p>
Sujet(s)
Animaux , Femelle , Rats , Densité osseuse , Vecteurs de médicaments , Epimedium , Chimie , Oestradiol , Sang , Flavonoïdes , Pharmacologie , Interleukine-6 , Sang , Ostéoporose , Sang , Traitement médicamenteux , Ovariectomie , Phospholipides , Phytothérapie , Rat Sprague-DawleyRÉSUMÉ
Aim The expression of NF-?B activation and the effect of antioxidant (N- acetylcysteine, NAC) on NF-?B activity in human airway epithelial cell was assessed. Methods Using the TNF-?,the airway epithelial cell strains of normal subject(16HBE) and tumor patient (H292) was activated and using Western-Blot and ELISA the expression of NF-?B and IL-8 were detected. Results It was found that the activity of NF-?B could be stimulated by the TNF-? and increase with the amount of TNF-? with the peak occurring at 2 to 4 hours after stimulation and then decreasing at six hours. At the same time, the level of IL-8 was elevated, but decreased with inhibition of NF-?B activity by NAC, that means the action of NAC has a dose-dependent effect. Conclusion NAC not only blocks the signal transmission activated by NF-?B, but also anticipates the transcription modulation of expression of many cell factors and inflammatory mediums. It suggests that NAC may play a role in the anti-inflammatory treatment of respiratory diseases.