PURPOSE: Recently, COMMD1 has been identified as a novel interactor and regulator of hypoxia-inducible factor-1 and nuclear factor kappa B transcriptional activity. The goal of this study was to determine the difference of COMMD1 expression in the placentas of women with normal and preeclamptic (PE) pregnancies. MATERIALS AND METHODS: Immnoperoxidase and immunofluorescent staining for COMMD1 was performed on nine normal and nine severe PE placental tissues, and COMMD1 mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: The expression of mRNA of COMMD1 was significantly higher in the study group than in the control group. The immunoreactivity was higher especially in the syncytiotrophoblast of PE placentas than in the control group. CONCLUSION: This study demonstrated increased placental COMMD1 expression in women with severe preeclampsia compared to that found in women with normal pregnancies, and this finding might contribute to a better understanding of the pathophysiology of preeclampsia.
Adaptor Proteins, Signal Transducing/genetics/isolation & purification/*metabolism ; Adult ; Female ; Humans ; Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy ; RNA, Messenger/metabolism

OBJECTIVETo screen proteins interacting with HCV NS4A protein in leukocytes by yeast-double hybridization.

METHODSThe bait plasmid pGBKT7-NS4A was transformed into yeast AH109 was transformed, and the expressing of the fusion protein was identified by SDS-page. The transformed yeast was mated with yeast Y187 containing leukocytes cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, analysis was conducted by bioinformatics. And, the gene encoding the interesting protein was cloned, and back-cross was performed.

RESULTSForty-five colonies were sequenced, among them, 29 colonies were human calcium modulating cyclophilin ligand (CAML). The gene encoding CAML was cloned, and the interaction between NS4A and CAML was ensured.

CONCLUSIONSeven kinds of proteins interacting with NS4A in leukocytes were successfully screened and the results brought some new clues for studying the pathogenesis of HCV.

Adaptor Proteins, Signal Transducing ; genetics ; isolation & purification ; metabolism ; Carrier Proteins ; genetics ; metabolism ; Cloning, Molecular ; Gene Library ; Humans ; Leukocytes ; cytology ; metabolism ; Protein Binding ; Transformation, Genetic ; Two-Hybrid System Techniques ; Viral Nonstructural Proteins ; Viral Proteins ; genetics ; metabolism

AIMAlthough epidermal growth factor receptors are expressed in the testes, whether they signal through epidermal growth factor receptor pathway substrate 8 (Eps8) is unknown. Here we evaluated the expression pattern of Eps8 in the maturing testis.

METHODSThe expression of Eps8 was analysed by Northern blotting, immunocytochemistry and Western blotting in primary Sertoli cell cultures and in testicular tissue of rodents.

RESULTSEps8 is specifically expressed in gonocytes, Leydig and Sertoli cells of the neonatal rats and in Leydig and Sertoli cells of the adult rats and mice. Although gonocytes express Eps8, no signal was found in prepubertal or mature spermatogonia and the expression level of Eps8 in Sertoli cells increases with age. No regulation of Eps8 expression in primary immature rat Sertoli cells by Follicle stimulating hormone (FSH) was detected by Western blotting.

CONCLUSIONEps8 seems to be involved in the growth factor-controlled regulation of cell proliferation and differentiation in the seminiferous epithelium. Eps8 is a possible marker for gonocytes and in Sertoli cells it could be involved in crosstalk with other growth factor pathways.

Adaptor Proteins, Signal Transducing ; Animals ; Base Sequence ; Blotting, Northern ; Cell Line ; DNA Primers ; Gene Expression Regulation, Developmental ; Immunohistochemistry ; Leydig Cells ; cytology ; physiology ; Male ; Proteins ; genetics ; RNA ; genetics ; isolation & purification ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; physiology ; Sexual Maturation ; Spermatocytes ; cytology ; physiology ; Testis ; growth & development