ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of LPXN overexpression on the proliferation, adhesion and invasion of THP-1 cells and its possible mechanism.</p><p><b>METHODS</b>A THP-1 cell line with stable overexpression of LPXN was constucted by using a lentivirus method, CCK-8 was used to detect the proliferation of cells, adhesion test was used to evaluate adhesion ablity of cells to Fn. Transwell assay was used to detect the change of invasion capability. Western blot was used to detect expression of LPXN, ERK, pERK and integrin α4, α5, β1, the Gelatin zymography was applied to detect activity of MMP2/MMP9 secreted by the THP-1 cells.</p><p><b>RESULTS</b>Successful establishment of THP-1 cells with LPXN overexpression (THP-1 LPXN) was confirmed with Western blot. THP-1 LPXN cells were shown to proliferate faster than the control THP-1 vector cells. Adhesion to Fn and expression of ERK, integrin α4, α5 and β1 in the THP-1 LPXN cells were higher than that in the control cells. Invasion across matrigel and enhanced activity of MMP2 could be detected both in the THP-1 LPXN cells as compared with the control cells.</p><p><b>CONCLUSION</b>Ectopically ovexpression of LPXN may promote proliferation of THP-1 cells through up-regulation of ERK; promote adhesion of THP-1 cells through up-regulating the integrin α4/β1 as well as integrin α5/β1 complex; promote invasion of THP-1 cells through activating MMP2.</p>
ABSTRACT
Objective To investigate the effects of simulated null gravity on the secretion of adrenomedullary hormones and miRNA-375 expression in rats. Method Fifty-six healthy adult male Wistar rats were randomly divided into 7 experimental groups (8 each), including suspension for 6h, 12h, 1d, 3d, 5d, 7d and 0h (control). Tail-suspension was used to establish the weightlessness animal model according to Morey-Holton's method. At the end of the experiment, the blood samples were obtained from the postorbitor vein, and the bilateral adrenal glands were harvested. The serum concentrations of catecholamines were determined by radioimmunological analysis. The expression of TH mRNA and miRNA-375 were assessed by quantitative RTPCR. TH protein in adrenal medulla was assayed using Western blotting analysis and immunohistochemistry. Results(1) The serum concentrations of rat's adrenomedullary hormones fluctuated obviously during the period of tail-suspension simulated weightlessness, with a temporary decline during 6-12h, followed by a sharp increase with a peak value on the 3rd day of suspension. (2) RT-PCR assay showed that TH mRNA expression in adrenal medulla of rats also fluctuated obviously, with a temporary decline at 6h and then elevated with a peak value at 1 day of suspension, followed by a downward trend forming a trough value on 5d of suspension. The immunohistochemistry showed that TH protein was stained as brown particles and the cytoplasm of adrenal medulla was stained deeply on 3d of suspension. (3) RT-PCR assay revealed that the expression of miRNA-375 in adrenal medulla increased temporarily at 6h of suspension, and it decreased significantly during 12h to 3d of suspension compared with that of control group (P<0.05). The miRNA-375 expression was up-regulated again during sustained suspension with a significant increase on 7d of experiment compared with that of control group (P<0.05). Conclusion The simulated weightlessness could affect the secretion of hormone of adrenal medulla in rats, and it may be related to the regulation of TH through miRNA-375 messenger network.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the genotype distribution of hemoglobinopathy in Chinese Jiangsu population.</p><p><b>METHOD</b>A total of 4115 samples were screened for hemaglobinopathy by using MCV combined with erythrocyte fragility tests and HPLC. Thalassemia genotypes were identified by Gap-PCR and Recerse Dot blot. PCR-DNA sequencing and PCR-elecrophoresis were used as supplement of PCR-RBD and for identifying the mutants of globin gene of abnormal hemoglobin.</p><p><b>RESULTS</b>The positive screening rate was 6.10% (251/4115) in Chinese Jiangsu population, 232 cases received thalassemia genotype diagnosis and from them 195 people were positive. In all positive ones, α-thalassemia, β-thalassemia, α-thalassemia combined with β-thalassemia, SEA-HPFH and SEA-HPFH combined with β-thalassemia were found respectively to be 31.28% (61/232), 66.15% (129/232), 1.54% (3/232), 0.43% (1/232) and 0.43% (1/232) of patients. The majority genotype of α-thalassemia was - - (SEA) and IVS-II-654 was the main genotype of β-thalassemia, 11 cases of abnormal hemoglobin were found, including 3 cases of Hb E, 1 Hb Kenitra, 1 Hb Seattle, 1 Hb Saitama, 1 Hb Bushwick, 1 Hb Koln and 1 Hb M-Milwaukee-2.</p><p><b>CONCLUSION</b>The main hemoglobinpathy is thalassemia in Chinese Jiangsu province and the HPLC play an important role in screening hemoglobinpathy. There is reference value of this study for genetic counseling and prenatal diagnosis.</p>