Purpose:To explore the expression of metastasis suppressor gene Kai1 in ovarian epithelial tumor and its significance.Methods:The expression of Kai1 protein in 108 samples of epithelial ovarian tumors and 12 of normal ovarian tissues was examined by S-P immunohistochemical staining.Results:The positive rates of Kai1 protein expression among malignant,borderline,benign and normal tumors were 26.6%,71.4%,73.3% and 83.3% respectively. The Kai1 protein expression in malignant tumors was obviously less than that in benign ones ( P 0.05). However,there was significant difference in Kai1 protein expression between samples of ovarian epithelial cancers with and without lymphonode metastasis ( P

AIM: To investigate different intracellular concentration of Ca 2+ in uterine myometrial cells at term and non-term.METHODS: The living cells suspensions were made to measure intracellular Ca 2+ concentrations after stainned by calcium fluorescent indicator Fluo-3 AM, then examined by laser scanning confocal microscope (LSCM).RESULTS: Intracelluar Ca 2+ showed very stronger red positive signal in myometrial cells at term than that in non-term cells. [Ca 2+ ]i were (35?8.1) nmol/L at non term and (75?7.3) nmol/L at term, which had significant difference compared with each other ( P

Objective Our purpose in this study is to investigate genes involved in the development of diabetes-induced embryonic malformations. Methods Two groups of 70-90 day old Sprague-Dawley rats were employed in our study: group 1 was normal control rats receiving a normal diet (n=3); group 2 consisted of experimentally-induced diabetic rats by intravenous injection of 65mg/kg of streptozotocin(STZ) on pregnancy day 6 with an attempt to reproduce malformations in embryos (n=3). Embryos were examined on day 12 under light microscopy to look for morphological defect of the neural tube (NTD). Yolk sac cells were harvested from each group and RNA was isolated. Genes expression profiles in yolk sac cells were analyzed using a DNA microarray technique. Results Gene expression patterns were compared in a total of 1200 genes between experimentally-induced diabetic rats and normal control rats, and 79 of genes were found to express differently between the two groups. Forty-two of genes were up-regulated in yolk sac cells of diabetic rats, such as apoptosis related genes BAX, bcl-2, heat shock 70kD protein and glucose-transporter 3; 37 of genes were down-regulated, such as phospholipase A2, insulin-like growth factor II receptor. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanisms underlying the developmental process during diabetes-associated embryonic morphogenesis, and it also might provide a useful tool in rapid diagnosis and prevention of malformation in early gestation stage of diabetic subjects.

Objective:To study the inhibitory effect of clarithromycin on angiogenesis induced by b-FGF. Methods:TheMatrigel implant assay was used. Matrigel (500?l) containing b-FGF and heparin were injected subcutaneously into the ab-domen of mice and were harvested 5 d later. The amount of hemoglobin and micro-vascular area present in the implant weremeasured and cotnpared. The mice were given either CAM (study group) or the same volume of glucose (vehicle group) oncea day by gastric intubation. Treatmen started 3 d before Matrigel implant and continued until the end of study. Results:Clar-ithromycin reduced hemoglobin content and micro-vascular area in Matrigel implant at high dosage(≥40 mg/(kg?d). Con-clusion:These data demonstrate that clarithromycin is a potent inhibitor of angiogenesis and may has possible therapeutic value in controlling pathologic angiogenesis.

Objectvies:To investigate the role of vascular endothelial growth factor(VEGF) in the pathogenesis of endometriosis(EMs). Methods:A total of 70 specimens of endometriosis and 30 specimens of the normal controls were evaluated by immunohistologically techique with polyclonal antibody against VEGF. Results:The expression levels of VEGF in endometriotic tissues was higher than that in the normal endometrium( P

Background and purpose: Recently, survivin gene is one of the hot spots in tumor study. The purpose of the present study was to observe the impact of RNAi targeting survivin gene on tumor growth, apoptosis and radiosensitivity in nude mice xenograft of human cervical carcinoma. Methods: HeLa-s2, HeLa-NC, HeLa-U6 neo and HeLa cells were inoculated respectively in flank subcutaneous tissue of 24 female nude mice so as to establish xenograft models of human cervical carcinoma randomly. The tumor growth status was observed. The tumor volume was regularly measured and the tumor weight was investigated used to observe the impact of RNAi targeting survivin gene on tumor growth. The expression of survivin protein and FⅧRAg in tumor tissues were examined by immunohistochemistry SP method MVD was calculated. Cell apoptusis in tumor tissues was observed by HE staining,apoptotic index(AI) was quantified by TUNEL method. To observe the impact of RNAi targeting survivin gene on tumor radiosensitivity after irradiated, the tumor growth delay and tumor weight were investigated and cell apoptosis were examined by TUNEL method. Results: We established 4 groups of xenograft models of human cervical carcinoma.The tumor volume of HeLa-s2 group was obviously less than that of HeLa group at every checkpoint. The tumor weight of HeLa-s2 group was obviously lower than that of HeLa group, and they were(0.369±0.043)g and (1.150±0.136)g respectively(P<0.05).The tumor growth inhibitive rate of HeLa-s2 group was 67.9%. The expression of survivin protein FⅧ RAg examined by immunohistochemistry in tumor tissues of HeLa-s2 group decreased sharply and MVD went down to 23.4±3.1. Apoptotic cells increased in tumor tissues of HeLa-s2 group examined by HE staining and TUNEL method, AI was (22.74±1.4)%. The tumor volume of HeLa-s2 group was obviously less than that of HeLa group at every checkpoint after irradiation and the tumor weight of HeLa-s2 group was obviously lower than that of HeLa group, they were:(0.41±0.06)g and(1.38±0.29)g respectively(P<0.05). Cell apoptosis increased in tumor tissues of HeLa-s2 group.Compared with HeLa group, AI of HeLa-s2 group rose up notably, they were(30.06±0.98)% and (4.17±0.64)%(P<0.05).Conclusion: RNAi for survivin gone can reduce MVD in xenograft by inhibiting survivin protein expression, thus inhibit tumor growth and induce cell apoptosis, Survivin gene RNAi can also enhance tumor radiosensitivity.

Objective To investigate the differently expressed genes in human ovarian carcinoma, and to reveal the molecular mechanism of the cancerous development. Methods The specimens of human ovarian serous adenocarcinoma tissues in stage III, and of normal human ovarian tissues as control were excised during surgery for present study. Clinical stages were determined by the Federation International of Gynecology and Obstetrids (FIGO). Total RNA was isolated from human ovarian tissues, and cDNA probe was labeled and purified. The amount of radioactivity incorporated into the cDNA probes was checked by a scintillation counter. The profiles of gene expression were compared between carcinomas and normal ovarian tissues by cDNA microarray which contained 588 genes totally. Results Forty-four differentially expressed genes were identified from the 588 genes which were from ovarian carcinoma and normal ovarian tissues and compared with cDNA expression array and analyzed by AtlasImage 1.01 software. 11 of the 44 genes were up-regulated in ovarian carcinoma tissues (including c-erbB2, neu, c-fos, c-myc proto-oncogenes, HER2 receptor, and so on), and the other 33 genes were down-regulated (including RAR, MMP18, MMP19, p21, DNA-PK, and so on). Conclusion The gene expressions in human ovarian carcinoma have been detected in present study. It is the differently expressed genes that help us to disclose the potential molecular mechanisms of the developmental process of human ovarian carcinogenesis. The differently expressed genes may provide a useful hallmark for the early diagnosis of human ovarian carcinoma.

Objective:To construct a short hairpin RNA(shRNA) eukaryotic expression vector of the survivin gene,and investigate its inhibitory effect on the survivin expression in human cervical cancer cells.Methods:We designed and synthesized 2 pairs of survivin-specific small interfering RNA primers(s1 and s2),cloned them into the eukaryotic expression vector pSilencer 2.1-U6 neo by DNA recombined techniques after annealing connection reaction,and transfected them respectively into human cervical cancer cell line HeLa using LipofectAMINE2000 after identificationby restrict endonuclease digestion and DNA sequencing.Then we selected the positive clones by G418 and detected the expression of survivin mRNA by semi-quantitative RT-PCR and that of the survivin protein by Western blot.Results:The survivin shRNA eukaryotic expression vectors pSilencer2.1-s1 and pSilencer2.1-s2 were successfully constructed.Positive clones were obtained by screening with G418 for 24 days,the survivin expression in HeLa cells decreased in different degrees after transfected with pSilencer2.1-s1 and pSilencer2.1-s2,and the latter showed a better interference efficiency.Conclusion:The shRNA eukaryotic expression vector of the survivin gene we constructed,with its improved interfering efficiency,has paved the way for further research on its role in regulating the biological behavior of cervical cancer cells.

OBJECTIVETo optimize the methods for genetic detection and prenatal diagnosis of Duchenne muscular dystrophy (DMD).

METHODSDenaturing high-performance liquid chromatography (DHPLC), multiplex PCR (mPCR), sequencing and other molecular techniques were used in combination for molecular diagnosis of 8 cases diagnosed as DMD.

RESULTSAmong the 8 cases, 4 have carried large deletions, 3 have point mutations, among which 6 were of de novo type. Prenatal diagnosis were offered for 5 families, the results showed that none of the fetuses had carried large deletions or point mutations. The pregnancies had continued and healthy babies were born.

CONCLUSIONCombined use of short tandem repeat, DHPLC, mPCR and sequencing can improve the detection of DMD gene mutations. By establishing and optimizing genetic and prenatal diagnostic methods, accurate genetic counseling can be provided for families affected with DMD.

Adult ; Base Sequence ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Testing ; Humans ; Molecular Sequence Data ; Muscular Dystrophy, Duchenne ; diagnosis ; embryology ; genetics ; Pedigree ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; Sequence Deletion ; Young Adult