RESUMO
OBJECTIVE: To examine the expression of gremlin-1 (GREM1) on the levels of messenger RNA (mRNA) and protein in eutopic endometrium and its serum level in patients with endometriosis. DESIGN: Prospective, experimental study using reverse-transcription polymerase chain reaction, Western blot, immunofluorescence, and ELISA. SETTING: Gynecological oncology laboratory in a department of obstetrics and gynecology in a medical college in China. PATIENT(S): Thirty-five patients with endometriosis and 23 healthy control women. INTERVENTION(S): During surgery, the eutopic endometria and peripheral serum were obtained from the patients with endometriosis and the control women. MAIN OUTCOME MEASURE(S): The cellular compartment location of GREM1 expression was examined by using immunofluorescent double staining. The expression levels of mRNA and protein for GREM1 were determined by reverse-transcription polymerase chain reaction and Western blot, respectively. The serum level of GREM1 was measured by indirect ELISA. RESULT(S): The expression of GREM1 was defined within endometrial blood vessel endothelium exclusively, with the concomitant expressions of GREM1 and CD146. The expression of GREM1 on the levels of mRNA and protein was significantly higher in eutopic endometria of patients with endometriosis than in those from healthy control women. According to the ELISA established in our laboratory, the concentration of GREM1 in peripheral serum that was collected during the follicular menstrual phase of patients with endometriosis was significantly higher than that in serum from healthy control women. CONCLUSION(S): Gremlin-1 plays a role to some extent in the aberrant angiogenesis of eutopic endometrium in patients with endometriosis. It is possible that the peripheral serum level of GREM1 is a prospective serum biomarker of endometriosis.
Assuntos
Endometriose/metabolismo , Endométrio/química , Peptídeos e Proteínas de Sinalização Intercelular/análise , Adulto , Biomarcadores/análise , Far-Western Blotting , Antígeno CD146/análise , Estudos de Casos e Controles , Endometriose/fisiopatologia , Endométrio/irrigação sanguínea , Ensaio de Imunoadsorção Enzimática , Feminino , Imunofluorescência , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Peptídeos e Proteínas de Sinalização Intercelular/genética , Neovascularização Fisiológica , Projetos Piloto , Estudos Prospectivos , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaRESUMO
A rapid and sensitive colloidal gold immunochromatography test strip based on one monoclonal antibody with broad-specificity, which can detect 12 fluoroquinolones (FQs), was developed. Antigen and goat anti-mouse IgG were respectively drawn on NC membrane as test line and control line. Gold-labeled antibody was added on a pad and put on one end of the membrane. Fluoroquinolones in sample solution compete with antigen combined on NC membrane for the gold-labeled antibody. When enough fluoroquinolone exists, the test line vanishes as there are no spare gold-labeled antibodies that can bind with antigen on the membrane. The control line always exists when the antibody is activated. The lowest detection limits of the FQs in spiked chicken muscle and chicken liver samples were 25 ng mL(-1) for norfloxacin and pefloxacin. The lowest detection limit for the other 10 FQs (enrofloxacin, ciprofloxacin, norfloxacin, flumequine, pefloxacin, ofloxacin, lomefloxacin, enoxacin, danofloxacin, amifloxacin, oxolinic acid, and marbofloxacin) was 50 ng mL(-1). The whole process involving sample preparation and detection can be finished in <10 min. The results demonstrate that the developed method can be potentially used as a screening tool for the determination of 12 FQ residues in a large amount of samples on site.