RESUMEN
OBJECTIVE: To study the effect of estrogen on anovulatory dysfunctional uterine bleeding (ADUB). METHODS: Primary endometrial epithelial cells of Hainan Lizu female was cultured and hydrolytic activity of gelatinase was determined by gelatin zymography analysis. Cellular mRNA and protein synthesis was blocked respectively to determine whether the increased expression of MMP-2/9 was induced by estrogen. The expression of VEGF was blocked by siRNA. After treatment with various factors, MMP-9, VEGF, total Erk and phosphorylated Erk expression in primary uterine epithelial cells was detected by Western blotting analysis. Cell MMP-2/9mRNA levels was measured by real-time RT-PCR. RESULTS: The activity and expression of MMP2/9 was increased in the endometrium of patients with ADUB. Estrogen could up-regulate the expression of VEGF and activate Erk 1/2-Elk1 signal path. After interference by siRNA, ERK1/2 pathway was blocked in cells, and the expression of MMP-2/9 was down-regulated. ERK1/2 specific blocker U0126 blocked ERK phosphorylation, and it could down-regulate the expression of MMP-2/9. CONCLUSIONS: The results showed that the estrogen can increase the expression of VEGF, and thus activate ERK1/2 pathway to induce MMP-2/9 expression.
Asunto(s)
Endometrio/citología , Células Epiteliales/enzimología , Estrógenos/metabolismo , Sistema de Señalización de MAP Quinasas , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Metrorragia/genética , Regulación hacia Arriba , Células Cultivadas , Endometrio/enzimología , Endometrio/metabolismo , Células Epiteliales/metabolismo , Femenino , Humanos , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Metrorragia/enzimología , Metrorragia/metabolismoRESUMEN
Inherited factor VII (FVII) deficiency is a rare disorder characterized by a bleeding phenotype varying from mild to severe. To date, more than 200 mutations have been described along the F7 gene encoding for FVII. The aim of this study was the identification of genetic defects underlying FVII deficiency in 10 patients belonging to eight unrelated families of the North provinces from Tunisia. Mutation detection was performed by sequencing the whole F7 gene coding region, exon-intron boundaries and about 400 bp of the promoter region. We identified 5 mutations in five unrelated families; the novel p.F328Y mutation and the reported mutations: p.R304Q, p.M298I, IVS1aG > A and p.G-39G. For the remaining 5 patients we didn't identified any mutations using PCR/Sequencing protocol. In conclusion, this study represents the first comprehensive molecular series of FVII deficiency affected patients in Tunisia from the North. We will try in the future to continue the molecular study for Tunisian patients from Center and South provinces in order to have a complete idea about the FVII deficiency mutational profile in our country. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1288044089753085.
Asunto(s)
Coagulación Sanguínea/genética , Deficiencia del Factor VII/genética , Factor VII/genética , Mutación , Polimorfismo de Nucleótido Simple , Adolescente , Adulto , Coagulación Sanguínea/efectos de los fármacos , Factores de Coagulación Sanguínea/uso terapéutico , Coagulantes/uso terapéutico , Contusiones/sangre , Contusiones/genética , Análisis Mutacional de ADN , Epistaxis/sangre , Epistaxis/genética , Exones , Deficiencia del Factor VII/sangre , Deficiencia del Factor VII/tratamiento farmacológico , Deficiencia del Factor VII/epidemiología , Femenino , Predisposición Genética a la Enfermedad , Humanos , Intrones , Masculino , Menorragia/sangre , Menorragia/genética , Metrorragia/sangre , Metrorragia/genética , Persona de Mediana Edad , Fenotipo , Regiones Promotoras Genéticas , Túnez/epidemiología , Adulto JovenRESUMEN
CONTEXT: Endometrial breakdown during menstruation and dysfunctional bleeding is triggered by the abrupt expression of matrix metalloproteinases (MMPs), including interstitial collagenase (MMP-1). The paracrine induction of MMP-1 in stromal cells via epithelium-derived IL-1alpha is repressed by ovarian steroids. However, the control by estradiol (E) and progesterone (P) of endometrial IL-1alpha expression and bioactivity remains unknown. OBJECTIVE AND DESIGN: Variations of endometrial IL-1alpha mRNA and protein along the menstrual cycle and during dysfunctional bleeding were determined using RT-PCR, in situ hybridization, and immunolabeling. The mechanism of EP control was analyzed using culture of explants, laser capture microdissection, and purified cells. Data were compared with expression changes of IL-1beta and IL-1 receptor antagonist. RESULTS: IL-1alpha is synthesized by epithelial cells throughout the cycle but E and/or P prevents its release. In contrast, endometrial stromal cells produce IL-1alpha only at menses and during irregular bleeding in areas of tissue breakdown. Stromal expression of IL-1alpha, like that of MMP-1, is repressed by P (alone or with E) but triggered by epithelium-derived IL-1alpha released upon EP withdrawal. CONCLUSIONS: Our experiments in cultured endometrium suggest that IL-1alpha released by epithelial cells triggers the production of IL-1alpha by stromal cells in a paracrine amplification loop to induce MMP-1 expression during menstruation and dysfunctional bleeding. All three steps of this amplification cascade are repressed by EP.
Asunto(s)
Endometrio/metabolismo , Epitelio/efectos de los fármacos , Hormonas Esteroides Gonadales/farmacología , Interleucina-1alfa/metabolismo , Menstruación/metabolismo , Metrorragia/metabolismo , Células del Estroma/metabolismo , Células Cultivadas , Endometrio/efectos de los fármacos , Epitelio/metabolismo , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Proteína Antagonista del Receptor de Interleucina 1/análisis , Proteína Antagonista del Receptor de Interleucina 1/genética , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Interleucina-1alfa/análisis , Interleucina-1alfa/genética , Interleucina-1beta/análisis , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Menstruación/genética , Metrorragia/genética , Modelos Biológicos , Técnicas de Cultivo de Órganos , Ovario/metabolismo , Comunicación Paracrina/genética , Comunicación Paracrina/fisiología , ARN Mensajero/metabolismo , Células del Estroma/efectos de los fármacosRESUMEN
OBJECTIVE: To research the expression of apoptosis related genes Bcl-2 and Bax protein in dysfunctional uterine bleeding. METHODS: Detecting the expression of Bcl-2 and Bax protein in 40 cases of endometrium tissue of dysfunctional uterine bleeding patients with immunohistochemistry antibiotics protein-peroxide emzyme (SP)methods. RESULTS: (1) The expression of Bcl-2 protein changes clearly and periodically in endometrium in normal cycle, the difference is obvious (P < 0.05). (2) The Bcl-2 protein develops with the hyperplasia in endometrium and the expression intensity is enhanced, the difference is obvious( P < 0.05). (3) The expression of Bax gene is masculine in endometrium in normal menstrual cycle. (4) The expression of Bax gene descends gradually with hyperplasia in uterine endometrium (P < 0.05). CONCLUSION: Apoptosis of endometrium cell is restrained from the excessive expression of Bcl-2 protein and deficient expression of Bax protein, so the uterine endometrium gets to hyperplasia or notypical hyperplasia.
Asunto(s)
Apoptosis/fisiología , Endometrio/metabolismo , Hiperplasia/patología , Metrorragia/patología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Neoplasias Uterinas/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Femenino , Expresión Génica , Humanos , Hiperplasia/metabolismo , Metrorragia/genética , Metrorragia/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Neoplasias Uterinas/patologíaRESUMEN
A congenitally abnormal fibrinogen was isolated from the blood of a young woman with a severe bleeding diathesis. Coagulation tests showed a prolonged Thrombin and Reptilase time partially corrected by Ca2+. Polymerization of thrombin induced preformed fibrin monomers was severely impaired. Thrombin caused the release of fibrinopeptides with normal retention times on HPLC. However, the rate of release was abnormally slow and the total amount of fibrinopeptide A (FpA) released reached only approximately 50% of the theoretical maximum. The rate and quantity of FpA release was normal when Reptilase was used. Transmission Electron Microscopy (TEM) of Thrombin induced clots showed an altered clot structure characterized by a reduced mean fiber diameter. The mother has a polymerization defect similar to the propositus, her fibrinopeptide release is unaffected however. The father has a minor fibrinopeptide release defect suggesting the presence of two populations of fibrinogen. This study supports the idea that the fibrinogen isolated from the propositus has two defects inherited as separate genetic traits. This fibrinogen has been named Fibrinogen Guarenas I.
Asunto(s)
Afibrinogenemia/genética , Fibrinógenos Anormales/aislamiento & purificación , Trastornos Hemorrágicos/genética , Adolescente , Biopolímeros/metabolismo , Femenino , Fibrina/metabolismo , Fibrina/ultraestructura , Fibrinógenos Anormales/genética , Fibrinógenos Anormales/metabolismo , Fibrinopéptido A/metabolismo , Fibrinopéptido B/metabolismo , Humanos , Cinética , Masculino , Metrorragia/genética , Microscopía Electrónica , Trombina/farmacología , Tiempo de TrombinaRESUMEN
Carriers of haemophilia A were recently described to have significantly more bleeding symptoms than healthy females. Eighteen obligate carriers and 20 females classified as carriers with approximately 95% confidence, as described in an accompanying paper, were compared with 56 noncarriers with respect to ten bleeding symptoms, registered by a questionnaire. A rank-order of negative findings indicating noncarrier state is presented, as well as a rank order of positive findings indicating carrier state. A summation index of the total diagnostic capacity of the ten questions shows that carriers have more bleeding symptoms than healthy females.