ABSTRACT
<p><b>AIM</b>To investigate the effect of iguratimod (T-614), a non-steroidal anti-inflammatory drug, on TNFalpha mRNA expression and TNFalpha production, and on the activity of nuclear factor-kappaB (NF-kappaB) in the rat alveolar macrophage cell line (NR8383) activated by LPS.</p><p><b>METHODS</b>NR8383 cells were pretreated with T-614 (13.4, 26.7, 53.4 micromol x L(-1)), then were stimulated with LPS. The production of TNFalpha in the supernatant of NR8383 was assayed by enzyme-linked immunosorbent assay (ELISA). The TNFalpha mRNA level was determined by a semi-quantitative PCR assay. Assessment of the NF-kappaB DNA binding activity was performed by an ELISA kit.</p><p><b>RESULTS</b>T-614 inhibited LPS-stimulated mRNA expression and production of TNFalpha in a concentration-dependent manner, as well as the activity of NF-kappaB. The IC50 value of effect of T-614 on TNFalpha level was 26.2 micromol x L(-1).</p><p><b>CONCLUSION</b>The inhibitory effect of T-614 on the production of TNFalpha in LPS-stimulated NR8383 cells may be mediated by suppression of NF-kappaB activity.</p>
Subject(s)
Animals , Rats , Anti-Inflammatory Agents, Non-Steroidal , Pharmacology , Cell Line , Cell Proliferation , Chromones , Pharmacology , Lipopolysaccharides , Macrophages, Alveolar , Metabolism , NF-kappa B , Metabolism , RNA, Messenger , Genetics , Sulfonamides , Pharmacology , Tumor Necrosis Factor-alpha , GeneticsABSTRACT
<p><b>BACKGROUND</b>We investigated the expression and role of TN4 in the oncogenesis of human hepatocellular carcinoma (HCC) from Qidong which is a HCC risk area.</p><p><b>METHODS</b>The expression of TN4 in HCC was observed using immunohistochemical staining (IHC). TN4 levels were manipulated in human liver cancer cell SMMC7721, using pcDNA3.1 eukaryotic expression constructs designed to express the complete TN4 cDNA. The biological changes of the cells were observed before and after transfection of TN4 and the change of gene expression was analysed by atlas cDNA expression array.</p><p><b>RESULTS</b>Among 100 pairs of samples of HCC, TN4 down-regulation expression and up-regulation expression positive rate were 81% (81/100), 19% (19/100), respectively (P < 0.01). TN4 protein was mainly localized in cytoplasm and membrane. The positive rate of TN4 were 10% (3/30), 100% (70/70) in lymph node metastasis and no lymph node metastasis, respectively (P < 0.01). The growth rates of the derivative SMMC7721-TN4 cell lines were decreased in comparison with that of normal SMMC7721 cells and pcDNA-SMMC7721. Some gene expression was changed before and after transfection of TN4. At 30 days of post-implantation of SMMC7721-TN4, SMMC7721-pcDNA3, SMMC7721 group produced tumors of (301.9 +/- 143.4) mm(3), (2418.7 +/- 362.8) mm(3), (2317.4 +/- 587.8) mm(3), respectively, (P < 0.01). Tumor inhibiting rate was 82.4% in TN4 transfection group. Sections of tumors were observed for their degree of tissue necrosis and there was higher degree of necrosis in tumors of the TN4-SMMC7721 cell group than those of the SMMC7721, SMMC7721-pcDNA groups.</p><p><b>CONCLUSIONS</b>TN4 may play an important role in the oncogenesis of human HCC, especially in Qidong, the HCC risk area and TN4 could be a candidate tumor suppressor gene for HCC.</p>
Subject(s)
Adult , Aged , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Epidemiology , Genetics , Carrier Proteins , Genetics , China , Epidemiology , DNA, Complementary , Gene Expression , Genes, Suppressor , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Liver Neoplasms , Epidemiology , Genetics , Lymphatic Metastasis , Genetics , Membrane Proteins , Genetics , Myelin Proteins , Nogo Proteins , Risk Factors , TransfectionABSTRACT
<p><b>OBJECTIVES</b>In order to identify whether the hens immunized with recombinant human testis prostaglandin D synthase (rhtL-PGDS) DNA can produce anti-L-PGDS antibody.</p><p><b>METHODS</b>The serum were got from the hens immunized with recombinant plasmid pGEX-2T/htL-PGDS DNA (100 micrograms) every 2 weeks for 2 times. The exist of anti-L-PGDS antibody and its titer were tested with agarose dual immunodiffusion and ELISA with rhtL-PGDS as antigen.</p><p><b>RESULTS</b>The serum anti-L-PGDS antibody in hen immunized with pGEX-2T/htL-PGDS DNA were confirmed and its titer tested by ELISA was 1:2,048.</p><p><b>CONCLUSIONS</b>It is feasible to produce anti-L-PGDS antibody by immunizing hens with recombinant pGEX-2T/htL-PGES DNA.</p>
Subject(s)
Animals , Female , Humans , Male , Antibodies , Chickens , DNA, Recombinant , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Genetic Vectors , Immunization , Intramolecular Oxidoreductases , Genetics , Allergy and Immunology , Lipocalins , Models, Animal , Plasmids , Genetics , Testis , Vaccines, DNA , Allergy and ImmunologyABSTRACT
Sperm must be capacitated before sperm-ovum fusion. Capacitation was once considered as hyperactivation. But now many investigators thought that capacitation wasn't equal to hyperactivation, and that sperm hyperactivation might be a moiety of capacitation or the result of capacitation. In the present, the methods used to study sperm capacitation include fertilization in vitro, induction of sperm acrosome reaction, FITC-labeled chlortetracycline and plant hemoagglutinin. The studies on sperm capacitation in vitro mainly focused on the inductive substances of sperm capacitation and subsequent results analysis. It could lay foundation for the manifestation of molecular mechanism of sperm capacitation and destination of sperm capacitation in molecular levels.