ABSTRACT
BACKGROUND: Previous studies have demonstrated that LRP16 is an estrogen-responsive gene. Its expression level is strongly associated with the proliferation and invasive growth of human breast cancer cells.OBJECTIVE: To construct a LRP16 targeting vector and screen mouse embryonic stem cell clones with homolougous recombination of an inactive LRP16 gene.DESIGN: Constructing an inserting inactivation target by inserting SA-RIES-β geo expression cassette.SETTING: Bioregulatory Laboratory of the Third Medical Department of Kyushu University in Japan and Department of Molecular Biology, General Hospital of Chinese PLA.MATERIALS: The materials used here were mainly provided by the Bioregulatory Laboratory, the Third Medical Department of Kyushu University in Japan. The mouse genomic library in pBeloBAC11 Vector was purchased from lnvitrogen Corp. The competent TopF10 was purchased from Beijing Tiangen Biotech Corp. pcDNA3.1(+) vector was kept in our laboratory. Mouse ES cells were provided by Kyushu University.METHODS: The experiment was performed in Kyushu University and Department of Molecular Biology of PLA General Hospital from November 2004 to May 2005. Targeting sequence of LRP16 gene was obtained from 129 mouse genomic Bacterial Artificial Chromosomes library based on polymerase chain reaction (PCR) screening. The SA-RIES-β geo fragment was inserted within LRP16 fifth exon to inactivate LRP16. ES cells were screened with G418 and the homologously recombinant clone was identified by Southern blot analysis.MAIN OUTCOME MEASURES: Clones with homologous recombination.RESULTS: The LRP16 fragment including exon 5 to 11 was subcloned into the pBluescript SK Ⅱvector. Restriction map demonstrated that the SA-IRES-β geo fragment was correctly inserted into the LRP16 fifth exon. Southern blot results showed that there was an ES clone with targeting sequence homologously inserted.CONCLUSION: A LRP16 gene targeting vector is constructed and a homologous recombinant is obtained.
ABSTRACT
Objective To investigate the morbidity, clinical characteristics, diagnosis, the pathological classification, character and special behaviour of transformation and the tendency of development of ovarian adenofibroma and cystadenofibroma, so as to provide information for clinical work. Methods 7 patients of ovarian adenofibroma and cystadenofibroma were retrospectively analysed on their onset age, clinical manifestations, ultrasonographic findings, pelvic examination and postsurgical pathology. Results All the seven tumors were serous. The ultrasonography revealed a cystic mass in the overy or near the overy, with clear boundary, and regular in shape. Three tumors were papillary (with rich signal of circulation in the papillary wall in two cases). In one case there was rich circulatory signal in the wall of the cyst. Three tumors had neither papilla on the wall of the cyst, nor blood signal. Pathological examination showed that 4 tumors were benign, and three were borderline. Conclusion The tumor is more prevalent in women of childbearing age. Unilateral tumor is more common than bilateral. The tumor is mainly serous, most of them are benign, and malignant tumors are rare. Ultrasonograply may be helpful to defining the nature and differentiation of the tumor