Spontaneous uterine perforation due to pyometra presenting as acute abdomen in older age / 대한산부인과학회지

Pyometra is an accumulation of purulent fluid in the uterine cavity. Generalized peritonitis secondary to a perforated pyometra is extremely rare. Most of the pyometra perforation is associated with malignancy. We have experienced a case of generalized peritonitis secondary to a perforated pyometra with no associated malignancy.

The Outcome of Pregnancy Combined with Idiopathic Thrombocytopenia Purpura and the Effect of Pregnancy on the Severity of This Disease / 대한주산의학회잡지

OBJECTIVE: The incidence of idiopathic thrombocytopenic purpura (ITP) is greatest in female during their childbearing years, so the concurrence of pregnancy and ITP is not unusual. Numerous studies have examined the outcomes of newborns, whereas fewer studies have been conducted with regard to the morbidity of obstetric patients with ITP. This study was aimed to find the outcome of pregnancy combined with ITP and the influence of the pregnancy on the severity of this disease. METHODS: From January 1996 to December 2005, a total of 62 pregnant women with ITP and their 73 deliveries were recruited for the study. Among them, 38 were diagnosed with ITP during pregnancy and the other 24 had pre-existing ITP before pregnancy. RESULTS: The severity of thrombocytopenia was exacerbated during pregnancy, but recovered to a level of non-pregnant period after delivery in most cases. The outcome of pregnancy of all the patients was uneventful except each one case of fetal demise at 35 gestational weeks and preterm delivery at 30 gestational weeks. One patient suffered from multiple subdural hemorrhage during pregnancy, which was spontaneouly recovered. Twenty newborns (27.8%) had transient congenital thrombocytopenia and 18 of them required treatment for hemostatic impairment. CONCLUSION: For women with ITP, Pregnancy can affect the severity of ITP, but life-threatening complication was almost lacking. Although, in not a few cases, there may need to treat both mothers and infants to raise their platelet counts, most mothers with ITP can proceed with their pregnancies and delivery healthy infant without complication.

Changes of Transfection Efficiencies of Plasmid LacZ Genes in Human Gynecologic Cancer Cell Lines / 대한산부인과학회잡지

OBJECTIVE: The transfection efficiencies of gynecologic cancer cell lines were investigated by different mediated transfection methods using recombinant LacZ plasmid (pRcCMVLacZ and pAAVCMVLacZ). METHODS: In this study, the gynecologic cancer cell lines were used CaSki, SiHa (cervical, HPV16+, wild type p53 gene), HeLa, HeLa S3 (cervical, HPV18+, wild type p53 gene), C33A, HT3 (cervical, HPV-, p53 mutant), HckE6/E7 (cervical, HPV16 immortalized keratocyte), PA-1 (ovary, wild type p53), SKOV-3, A2774 (ovary, p53del) and OVCAR-3 (ovary, p53 mutant). The pRcCMVLacZ and pAAVCMVLacZ plasmid transfection were performed by using liposome system such as Ca2+-phosphate, Fugen6(TM), Lipofection(TM), Lipogen(TM) and N-stearyl lactobionamide (N-SLBA) with X-gal staining. The LacZ gene was used the reporter gene for the transfection efficiencies evaluation. RESULTS: Each of cell lines were showed different transfection efficiencies by Ca2+-phosphate, Fugen6(TM), Lipofectin(TM), Lipogen(TM) and N-SLBA. Each of cell were revealed that HeLa S3, HT3 and A2774 were high transfection efficiency using the pRcCMVLacZ by the Lipogen(TM), SiHa, HeLa, QGU, OVCAR-3 and PA-1 were high efficiency using the pAAVCMVLacZ by Lipofectin(TM), CaSki was high efficiency using the pRcCMVLacZ by the Lipogen(TM), A2774 and Cx16.2 were high efficiency using the pRcCMVLacZ by the Lipofectin(TM), SKOV-3 and HkcE6/E7 were high efficiency using pAAVCMVLacZ by the Lipogen(TM). CONCLUSION: As a result, We proved that each of cell lines differed trasnfection efficiencies according to mediated transfection and recombinant LacZ plasmid style. Above all, Lipofectin(TM) mediated transfection was showed high efficiency at the most of cell lines.

Selective Protection of Normal Proliferationg Cells Against Anti-Neoplastic Chemotherapy by Stausporine Pretreatment / 대한산부인과학회잡지

OBJECTIVE: A major limiting factor in human cancer chemotherapy is toxicity in normal cells and tissues. Our goal was to determine whether normal proliferating cells could be protected from chemotherapeutic agents by taking advantage of the differential drug sensitivity of cell cycle G1 checkpoint in normal and cancer cells. METHODS: Normal peripheral blood mononuclear cells (PBMC) and ovarian cancer cell lines (OVCAR- 3 and SKOV-3) were initially treated with 10 nM of staurosporine for 48 hours. After removal of staurosporine contained media, both PBMC and ovarian cancer cells were treated with 20 nM of paclitaxel for 24 hours. Cells were then allowed to recover in drug-free medium for 4 days. The DNA contents and cell cycle changes were detected by FACScan flow cytometer in the cells harvested whenever the medium was changed. RESULTS: After pretreatment of ovarian cancer cell lines (OVCAR-3 and SKOV-3) with 10 nM of staurosporine followed by treatment with 20 nM of paclitaxel, both OVCAR-3 and SKOV-3 cells were selectively arrested in G2M phase of cell cycle by paclitaxel and they resumed their proliferative cycle to some extents after the drugs were removed and cultured with fresh media. However. pretreatment with 10 nM of staurosporine protected normal circulating PBMC that had been induced to proliferate in vitro with phytohemagglutinin from paclitaxel. Staurosporine-induced arrest of PBMC in G0/G1 phase was reversible, and arrested cells tolerated 10 nM of paclitaxel in culture. CONCLUSION: OVCAR-3 and SKOV-3 cancer cells can be targeted specifically with paclitaxel, following staurosporine-mediated, selective and reversible G0/G1 arrest in PBMC.

Flow Cytometric Detection of Apoptosis and p53 Protein in OVCAR-3 and SKOV-3 Ovarian Cancer Cell Lines / 대한산부인과학회잡지

OBJECTIVE: Taxol (paclitaxel)-induced apoptosis was studied to understand their biological mechanism correlated with the expression of p53 in the SKOV-3 and OVCAR-3 ovarian cancer cell lines. MATERIALS AND METHODS: The SKOV-3 and OVCAR-3 cell lines were cultured in RPMI 1640 medium without taxol (control group) and with taxol for 24 h and 48 h (experimental group). After harvest, the cells were stained with annexin V-FITC (fluorescein isothiocyanate) and anti-cytokeratin antibodies (clone CAM5.2 and clone MNF116). They were washed and stained with p53 antibody. After then the secondary antibodies, i.e., FITC- or phycoerythrin (PE)-conjugated goat anti-mouse (GAM) immunoglobulin G (GAM IgG-FITC or GAM IgG-PE) were added in the cells and they were incubated in the dark. DNA of these cells were stained sequentially with propidium iodide (PI). Standard FACScan equipped with a 488 nm single laser was used for the analysis of these cells. RESULTS: Both of SKOV-3 and OVCAR-3 cell lines were arrested in the G2M phase after treatment of taxol, suggesting that these cells would eventually enter into the stage of cell death. Fractions of negative cytokeratin and positive annexin V and amount of sub-G0G1 fraction indicative of apototic fractions were lower in the SKOV-3 cell line compared with that in OVCAR-3 cell line, probably as a result of lower sensitivity of SKOV-3 cell line to the taxol. p53 expression were not detected in SKOV-3 cell line. On the basis of observed findings in SKOV-3 cell line and findings of high expressions of p53 regardless of taxol treatment, no increases in their expressions according to culturing time, and gradual increases in sub-G0G1 fractions and in fractions of negative cytokeratin and positive annexin V indicative of apoptosis in OVCAR-3 cell line, we concluded that the expression of p53 would not be associated with cell cycle changes and the arrest in the G2M pahse but associated with the appearance of apotosis. CONCLUSION: Our results suggest that flow cytometric detection of the apoptotic fractions would be an effective, fast, and accurate method for the chemosensitivity test in tumor cells before the administration of anti-cancer drugs in gynecologic cancer patients.

Detection of p16(INK4A) in the Mixed Cell Populations of Normal Peripheral Blood Mononuclear Cells and Cervical Cancer Cell Lines / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Human papilloma viruses (HPVs) play a central role in the pathogenesis of neoplastic lesions of the uterine cervix. The viral oncoprotein HPV E6 degrades the p53 protein, and the HPV E7 protein inactivates pRB and increases the expression of the CDK inhibitor, p16(INK4A). We investigated the usefulness of p16(INK4A) as a biologic marker for the cervical dysplastic and neoplastic cells. MATERIALS AND METHODS: We examined the expression of p16(INK4A) and cytokeratin in a mixed population of normal peripheral blood mononuclear cells (PBMC) and the cervical cancer cell lines (HeLa, SiHa, and CasKi) using flow cytometry. RESULTS: The DNA indices of the HeLa, SiHa and CasKi cell lines were 1.89, 1.53 and 1.75, respectively, indicating that these cells are aneuploid cells. Furthermore, the positive rate of p16(INK4A) expression was 86.7% for the HeLa mixed population, 85.6% for the SiHa mixed population, and 92.2% for the CasKi mixed population. According to the FL3A vs FL3W histogram, electrical gating of the HeLa, SiHa and CasKi mixed populations showed the expression levels of both cytokeratin and p16(INK4A) to be identical, at 86.6%, 84.8% and 85.0%, respectively. These findings revealed that almost all cells selected through electrical gating were cervical cancer cells originating from the epithelium and which expressed cytokeratin and p16(INK4A). On the other hand, when each mixed population was electrically gated for normal PBMC, we found that the PBMCs expressed neither cytokeratin nor p16(INK4A). CONCLUSION: Using flow cytometry, we observed the enhanced expression of p16(INK4A) in cervical cancer cell lines. These RESULTS suggest the usefulness of p16(INK4A) for the selective detection of cervical dysplastic and cancer cells in the liquid-based samples, which are taken from the cervices and contaminated with blood and stromal cells.