ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of Gecko Swinhonis Gunther freeze-dried powder (GFP) in inducing apoptosis of C6 glioma cells.</p><p><b>METHODS</b>Thirty mice were randomly divided into three groups and treated with intraperitoneally injection of cisplatin, orally taken GFP or distilled water respectively. After treatment for 20 days, the serum of mice was collected for treating the C6 glioma cells. The apoptosis state of cells was observed by morphological examination, flow cytometry and TUNEL method, and the cell apoptosis related gene expression of bcl-2 and bax were determined by S-P immunocytochemical assay.</p><p><b>RESULTS</b>The C6 glioma cell apoptosis induction of GFP contained serum in vitro could be confirmed by morphological examination, flow cytometry analysis and TUNEL method. Comparison between the GFP treated group and the blank control group on intracellular bcl-2 gene expression showed no difference, but bax gene expression was higher in the GFP treated group.</p><p><b>CONCLUSION</b>GFP contained serum could induce C6 glioma cell apoptosis, its mechanism might be related with the up-regulation of bax gene.</p>
Subject(s)
Animals , Humans , Male , Mice , Antineoplastic Agents , Pharmacology , Apoptosis , Drugs, Chinese Herbal , Pharmacology , Glioma , Chemistry , Genetics , Pathology , Materia Medica , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Genetics , Random Allocation , Serum , Tumor Cells, Cultured , bcl-2-Associated X ProteinABSTRACT
Objective: To examine the alteration of pathologic structure and endogenous hydrogen sulfide pathway in rats with pulmonary hypertension induced by high pulmonary blood flow. Methods: Sixteen SD rats were randomly divided into shunting group and control group. An 11 week aortocaval shunting was produced in rats of shunting group, and pulmonary artery mean pressure (mPAP) was evaluated using right cardiac catheterization. The ratios of right ventricular mass to body weight (RV/BW) and right ventricular mass to left ventricular plus septal mass[RV/(LV+S)] were also detected. Pulmonary vascular micro and ultra structures were examined. Meanwhile the concentration of plasma hydrogen sulfide (H 2S) was measured by spectrophotography. The gene expression of cystathionine ? lyase (CSE)was detected by in situ hybridization, and the activity of CSE in lung tissues was measured by H 2S production according to chemical analysis. Results: After 11 weeks of aortocaval shunting, pulmonary artery mean pressure was significantly increased. Muscularization of small pulmonary vessels and relative medial thickness of pulmonary arteries were obviously increased in shunting rats compared with controls. Ultrastructure of intrapulmonary arteries changed obviously in shunting rats. Meanwhile, plasma H 2S concentration was decreased and the activity of CSE (according to H 2S production) in lung tissues decreased in shunting rats. CSEmRNA expression by pulmonary arteries was significantly suppressed. Conclusion: Pulmonary vascular structural remodeling is the important pathologic basis for pulmonary hypertension induced by high pulmonary blood flow. The down regula tion of endogenous H 2S pathway might play an im portant role in the development of high pulmonary blood flow induced pulmonary hypertension.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.</p><p><b>METHODS</b>The specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.</p><p><b>RESULTS</b>TCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.</p><p><b>CONCLUSION</b>The idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.</p>
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Anti-Idiotypic , Blood , Allergy and Immunology , Base Sequence , Complementarity Determining Regions , Genetics , Allergy and Immunology , Epitopes , Genetics , Allergy and Immunology , HL-60 Cells , Lymphoma , Allergy and Immunology , Mice, Inbred BALB C , Molecular Sequence Data , Receptors, Antigen, T-Cell, alpha-beta , Genetics , Allergy and Immunology , Sequence Analysis, DNA , Tumor Cells, Cultured , Vaccines, DNA , Genetics , Allergy and ImmunologyABSTRACT
] AIM: To investigate the expression of the urotensin Ⅱ (UⅡ) receptor GPR14 in the aorta of apoE knockout mouse. METHODS: The expression of GPR14 in the aorta of apoE knockout C57BL/6J mice at various ages (18 weeks, 28 weeks, and 38 weeks old, respectively) was determined with competitive RT-PCR. A binding assay of [ 125 I]-UⅡ on the aortic tissue was also performed in 28 weeks group. RESULTS: We found significant upregulation of GPR14 mRNA at all three ages. Compared with wild type group at the same age, the GPR14 mRNA level in apoE knockout mice increased 54.2% in 18 week group (P