Ovarian folliculogenesis is a dynamic process during which follicles undergo growth and differentiation. Recently, apoptotic programmed cell death has been suggested to be one of the underlying mechanisms responsible for the regulation of ovarian folliculogenesis. In the present study, effects of tumor necrosis factora (TNFa) and transforming growth factor/3 (TGF/3) on the proliferative potential, endocrine function, and apoptotic internucleosomal DNA fragmentation of granulosa cells were examined in vitro as a function of follicular growth using a porcine model. Porcine granulosa cells obtained from small (1-2mm), medium (3-5mm) and large (6-12mm) follicles were cultured under a serum-free condition in the presence or absence of FSH (100ng/ml) and IGF-I (100ng/ml), with or without various concentrations of TNFa or TGFfi. Proliferative activity of cultured granulosa cells was assessed by immunocytochemical techniques with a monoclonal antibody to proliferating cell nuclear antigen (PCNA). Endocrine function of the cells was assessed by determining the ability to secrete 17, 8-estradiol (E2) and progesterone (P4). Apoptotic internucleosomal DNA fragmentation of the cultured cells was analyzed by in situ DNA 3'-end labeling method with a digoxigenin-dideoxy-UTP. In the present study, FSH and IGF-I were used as a very important endocrine factor and local factor, respectively, for granulosa cell growth and function. Addition of FSH and IGF-I augmented proliferative activity and steroidogenic ability of cultured granulosa cells, and remarkably diminished internucleosomal DNA fragmentation of the cells. Concomitant treatment with TNFa or TGFQ decreased FSH and IGF-I-stimulated proliferative activity and E2secretion of cultured granulosa cells, but increased FSH and IGF-I-inhibited apoptotic DNA fragmentation of the cells. The inhibitory effects of TNFa and TGF, Q on the proliferative activity of cultured granulosa cells were prominent in small follicle granulosa cells, whereas the inhibitory effects of TNFa and TGF/3on E2 secretion were prominent in medium follicle and large follicle granulosa cells. Furthermore, TNFa decreased FSH and IGF-I-stimulated P4 secretion by cultured granulosa cells, whereas TGFf did not significantly alter P4 secretion by the cells. The inhibitory effect of TNFcr on P4 secretion was apparent regardless of the stage of follicular growth. The lag time required for the inhibitory effects of TNFa and TGF13 on the proliferative activity of cultured granulosa cells was 24h, while that for the inhibitory effects of TNFa and TGFf on the E2 secretion was 48h. These results suggest that the biological action of TNFa and TGF$ on granulosa cells may shift from the inhibition of the proliferative activity in immature follicles to the inhibition of the endocrine function in mature follicles during the course of follicular growth. It is also likely that FSH and IGF-I act as follicle survival factors whereas TNFa and TGFJ3 act as atretogenic factors in porcine granulosa cells during follicle development. [Adv Obstet Gynecol 49 (5); 507---515, 1997(H9.9)]

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The aim of this study is to clarify whether modification of the local milieu surrounding the embryo improves the implantation rate. It has been demonstrated that pregnancy can be achieved by the direct insertion of embryos into the endometrium of mice. To modify the local milieu surrounding the embryo, we inserted blastocysts together with various bioactive substances into the endometrium of pseudopregnant mice. The implantation rates were evaluated 8 days after embryo transfer. The substances tested were histamine, prostaglandinE2, progesterone and human chorionic gonadotropin. Among them, only histamine exerted the effect on the embryo implantation. The implantation rate was improved on day 2 (31.5 %) when embryos were transferred together with histamine (100pM). A study to clarify the mechanism of this improvement by histamine is now under way. So far, histamine has had no effect on blastocyst development in vitro in terms of trophoectoderm outgrowth or on decidualization in an endometrial stromal cell culture. Marked stromal edema was found around the embryos transferred with histamine. Histamine may enhance implantation not by directly affecting embryos and decidualization in the stroma but by modifying the local milieu surrounding embryos as a result of increased vascular permeability. [Adv Obstet Gynecol 49 (5); 516---527, 1997 (H9.9)]

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Growth of mouse preimplantation embryos in several kinds of coculture with somatic cells was observed and related embryotropic factors were investigated. Mouse pronuclear stage embryos were cocultured with rabbit oviductal cells, rabbit endometrial epithelial cells, human dermatofibrosarcoma cells, human liposarcoma cells, guinea-pig intraperitoneal macrophages, or mouse intraperitoneal macrophages, respectively. Mouse embryos grew in coculture with rabbit oviductal cells the same as in basal medium (control). Embryos cocultured with rabbit endometrial epithelial cells, human dermatofibrosarcoma cells or human liposarcoma cells showed significantly worse development than the control. Coculture with guinea-pig intraperitoneal macrophages or mouse intraperitoneal macrophages significantly enhanced embryonic development. Although the number of cells comprising the embryos significantly increased in coculture with MO, these were less than those of in-vivo embryos. Decrease of the number of cells of MO coexisting with embryos reduced the enhancing effect of embryonic development in coculture. Prolongation of time between harvest of MO and coculture brought less embryotropic effect. Embryos showed worse growth in conditioned medium of MO than the control. Although twenty five amino acids were detected in the conditioning medium with MO, the amounts were very low. In analysis with antibodies against human or rat materials, IL-1Q was 100 pg/ml and other cytokines or growth factors (IL-1a, IL-1/3, IL-6, TNF-a, IFN-a, TGF-a, TGF-f, IGF- I, IGF-II, EGF, PDGF-AB, PDGF-BB, GM-CSF, G-CSF) were undetectable. Prostanoids in the conditioned medium were quantified as follows; PGF2a: 325.0 pg/ml, PGE2: 635.0pg/ml, 6keto-PGF, a: 235. Opg/ml, TXB2: 2180.0pg/ml. Indomethacin had no influence upon coculture with M. Coculture with Mck did not improve the implantation rate in transfer to pseudopregnant mice. In conclusion, coculture with guinea-pig intraperitoneal macrophages improved embryonic development with the effect being thought to be brought about by an unstable factor other than cytokines, growth factors and prostanoids. The implantation rate of cocultured embryos was very low in spite of morphological improvement. It was suggested that although a system improved embryonic development morphologically, as did our coculture system, the pregnancy rate may not necessarily accelerate. [Adv Obstet Gynecol 49(5); 528-542, 1997 (H9.9)]

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Nausea, vomiting, and diarrhea are common gastrointestinal side effects of anticancer chemotherapy, usually managed with symptomatic treatment. Severe gastrointestinal side effects can occur. We report a ptient with ovarian cancer in whoni ischemic colitis began after treatment with cisplatin and granulocyte-colony stimulating factor (G-CSF). A 57-year-old woman with ovarian cancer in stage Ic underwent cisplatin chemotherapy after surgery. After two courses of treatment, she was given G-CSF and antibiotics because of severe myelosuppression. A third course of cisplatin was given after bone-marrow recovery. At this time, severe pain in the abdomen began and bloody stools were produced. The diagnosis was ischemic colitis, on the basis of findings by colonoscopy. The patient was managed successfully by conservative therapy alone. The etiology of ischemic colitis in this patient was probably multifactorial, and anticancer chemotherapy and G-CSF may have been pathogenic factors. Ischemic colitis caused by chemotherapy is uncommon, but it may require special care. [Adv Obstet Gynecol 49 (5); 543-554466, , 1997 (H9.9)]

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A case of gestational choriocarcinoma with lung and brain metastases was reported in this paper, and the complete remission was achieved by multidisciplinary treatments comprising chemotherapy, irradiation and surgery. The patient was a 27 year-old multiparous female. Her first pregnancy was a complete hydatidiform mole which required methotrexate single agent chemotherapy. It was followed by a missed abortion and two normal deliveries. She was referred to our hospital for evaluation of irregular menstruation and occasional abnormal vaginal bleeding for 6 months, and headache with vomiting for half a month. On admission, she had nystagmus, rigidity of the neck and swaying gait. There was no vaginal bleeding. The urinary hCG level was 115, 640 IU/day, and the serum free fl-hCG was 166ng/ml. Chest X-ray showed a round tumor of 10 cm in diameter in the left lower lung field. Brain computed tomography showed a round, hemorrhagic tumor of 3 cm in diameter in the right cerebellar hemisphere, pons herniation and hydrocephalus. Pelvic magnetic resonance imaging showed no pathological findings in the internal genitalia. Craniotomy was indicated due to signs of increased intracranial pressure. The resection of cerebellar tumor was performed. Postoperative pathological study revealed choriocarcinoma. EMA-CO combination chemotherapy and whole-brain irradiation was initiated at the first postsurgical week. The chemotherapy brought rapid decrease of both the urinary hCG level and size of the lung tumor. After the sixth course of the chemotherapy, left lower lobectomy with the partial resection of parietal pleura was performed and the urinary hCG level became undetectable. Three further courses of the chemotherpy were then administered. Finally, hysterectomy with left salpingo-oophorectomy and right salpingectomy was performed. At one year after discharge, she showed sign of neither recurrence, respiratory dysfunction nor neurological abnormality. [Adv Obstet Gynecol 49 (5); 547-553, 1997 (H9.9)]

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Recently, several applications of HDC (High Dose Chemotherapy) combined with PBSCT (Peripheral Blood Stem Cell Transplantation) to ovarian cancer treatment revealed in many hospitals. We had experienced three ovarian cancer cases treated with HDC combined with PBSCT. We estimated these cases about the appropriate agents combination for mobilization chemotherapy, the use of G-CSF (Granulocyte Colony Stimulating Factor) and the suitable time of peripheral blood stem cell harvest under monitoring of CD34 as a stem cell marker. High CD34 positive stem cell harvest ratio was obtained, when bone marrow suppression preceding induction chemotherapy was strong, G-CSF application was started at the point nadir of peripheral white blood cells and PBSCI-I was carried out at the time of stem cells mobilization into peripheral blood. [Adv Obstet Gynecol 49 (5): 554-562, 1997 (H9.9)]

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We encountered an extremely rare case of primary malignant melanoma of the cervix. A 72year-old female with multigravidity (twice) and multiparity (twice) consulted our hospital for genital bleeding. In the vaginal portion of the uterus, phyma with partial black pigmentation was noted. Cervical scraping smear revealed atypical cells with naked nuclei or omnipresent nuclei, marked increases in chromatin and little intercellular binding. The cytoplasm included brown granules. Clinical stage of the patient was classified as cervical cancer stage II b. Abdominal radical hysterectomy with bilateral salping000phorectomy and radiotherapy were performed. Histopathology revealed round tumor cells with polymorphism. In these cells, large nucleoli, intranuclear vacuoles, nuclear division and brown melanin granules with in the cytoplasm were detected. On immunohistochemistry, tumor cells were positive for HMB-45 and S-100 proteins. Electron microscopy revealed melanosomes in the cytoplasm. Sixth months postoperatively, she died because her disease continued to progress rapidly. [Adv Obstet Gynecol 49 (5); 563-567, 1997 (H9.9)]

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[Mitochondria] myopathy is caused by several biochemical defects involving the energy-producing mitochondrial enzyme system. The disease is associated with impairment of muscular function, which may be progressive, intermittent or fluctuant. Inheritance is thought to be via maternal transmission. MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) is a type of the mitochondrial myopathy characterized by stroke-like episodes, elevation of lactate level in both the blood and cerebrospinal fluid, and pathologically or biochemically abnormal finding of the mitochondria. We report a case of MELAS. The patient developed exercise intolerance at 27 weeks of gestation and had normal parturition and puerperium. She was a 31-year-old nulliparous woman who had two siblings diagnosed as MELLAASS.. At the age of 25, she had experienced seizure-like episodes. She complained of muscular hypotonia, exercise intolerance and uterine contraction at 27 weeks of gestation. Although periodical uterine contraction was shown by CTG, gynecological examination and sonography revealed no remarkable change. The disturbance of electrolytes, severe metabolic acidosis and elevated lactate level were noted by laboratory procedure. The presence of a T to C point mutation at nucleotide position 3271in the mitochondrial DNA tRNA`-`'"'1"' was indicated by the mitochondrial DNA analysis of the peripheral blood. The muscular hypotonia and exercise intolerance were gradually improved by supplement of electrolyte and adjustment of acid-base balance. At 38 weeks, she went into spontanous labor. Although labor progressed normally, vacuum extraction was performed due to non-reassuring FIIR pattern indicating severe variable deceleration at the second stage of labor. She delivered transvaginally a male infant weighing 3472 gram with Apgar score of 5 and 10 at 1 and 5 minutes. Although postpartum lactic acidosis required administration of 7 % sodium bicarbonate, the postpartum course for both the mother and infant was uneventful. Recently, it is well accepted that a mutation of mitochondrial DNA causes not only mitochondrial myopathy but diabetes mellitus, cardiomyopathy and other degenerative disease. Further investigation on perinatal care of pregnancy with mitochondrial disease are needed. [Adv Obstet Gynecol 49 (5); 568-572, 1997 (H9.9)]

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We encountered a patient with uterine polypoid adenomyoma associated with adenofibroma that has been rarely reported. When she underwent abdominal myomectomy, an irregular hyperplastic tissue mass accompanied by necrosis and polyps was observed in the uterine cavity. Histological examination during operation revealed a benign mixed tumor of epithelial and non-epithelial origin. However, cytological examination showed a few atypical tumor cells of non-epithelial origin, and simple total hysterectomy was perfomed. Resected uterine specimens demonstrated combination of benign tumors. Small multiple polyps of the uterine body showed endometrial glands and hyperplasia of fibroblasts and smooth muscle fibers, which were diagnosed to be adenomyoma by immunohistological staining using an anti-desmin antibody. The tumor of the endocervix was diagnosed to be adenofibroma with phyllode papillary proliferation consisting of cervical glands and fibroblasts. [Adv Obstet Gynecol 49 (5); 573---577.1997 (H9.9)]

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