ABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.</p><p><b>METHODS</b>Using combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.</p><p><b>RESULTS</b>Sertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).</p><p><b>CONCLUSION</b>The proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.</p>
Subject(s)
Animals , Male , Rats , Cells, Cultured , Glial Cell Line-Derived Neurotrophic Factor , Metabolism , Rats, Wistar , Sertoli Cells , Cell Biology , Metabolism , Temperature , Testis , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To study the differences in expression of nuclear factor-kappaB (NF-kappaB) between human traumatic cataract and normal lenticular epithelial cells.</p><p><b>METHODS</b>Total RNA of anterior capsule specimens was taken under the microscope from normal cadaveric eyes donors and those suffering from traumatic cataract to make semi-quantitative RT-PCR and conduct analysis of differences in expression of NF-kappaB between them.</p><p><b>RESULTS</b>As compared with the mean of 0.8337 in normal control group, the expression equivalent of NF-kappaB was 0.9074 for the lenticular epithelial cells in traumatic cataract sufferers, and the differences are of noticeable significance (t = 2.447, P<0.05) accordingly.</p><p><b>CONCLUSIONS</b>NF-kappaB is likely a kind of transcription factor necessary to maintain metabolism of normal lenticular epithelial cells. Higher NF-kappaB available in the traumatic cataract sufferer's lenticular epithelial cells means NF-kappaB is of possible relevance to occurrence and development of traumatic cataract.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Cataract , Genetics , Metabolism , Therapeutics , Epithelial Cells , Metabolism , Lens, Crystalline , Wounds and Injuries , Metabolism , NF-kappa B , Genetics , Metabolism , Polymerase Chain ReactionABSTRACT
Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P
ABSTRACT
Attention deficit hyperactivity disorder(ADHD)is a common childhood behavioral disorder,and it is characte-rized by the core symptoms of attention deficit,hyperactivity and impulsivity.In recent years,a variety of neuropsychological deficits related to ADHD have been confirmed,including intelligence,attention and inhibition,verbal and spatial working memory,set shift,planning and monitoring.Some important theories related to the metal mechanism of ADHD have been put forward,and the representatives include behavioral inhibition theory,cognitive-energetic model,dual pathway theory and state regulation theory.