Immunosuppressive effect of culture supernatant derived from decidual cell rich fraction

The immunosuppressive function of culture supernatant of decidual cell rich fraction (Dec. supp.) was studied with the aim of clarifying the role played by decidual tissue in maintaining pregnancy. To obtain decidual cells, decidual tissue from miscarriages was enzyme-treated and submitted to Percoll discontinuous specific gravity centrifugation in specific gravity fractions from 1,025 to 1,065 in steps of 0.01. Decidual cell rich fraction was obtained in specific gravity fractions at 1,035 and 1,045. The Dec. supp. was obtained by incubating the decidual cell rich fraction for 24 hours. The effects on cell-mediated immunity were studied with the PHA reaction of mononuclear cells (MNC) and the one-way mixed lymphocyte reaction (MLR) among normal subjects. For the PHA reaction, suppression was 19.5 +/- 7.8% and 38.2 +/- 8.9% after the administration of Dec. supp. at 20% and 50% respectively. For the MLR, conspicuous suppression of 48.0 +/- 19.1% and 61.0 +/- 31.2% was seen after administration at 20% and 50% respectively. Next, indomethacin (ind.), a synthetic suppressor of prostaglandin (PG), was added in order to determine what part PG in the culture supernatant plays as one of the immunosuppressive factors in Dec. supp. Suppression was notably weaker. The PGE density in Dec. supp. was measured and found to be as high as 840 +/- 350 pg/ml, which suggests the possible intervention of PG as one of the suppressing factors.     

Growth-inhibitory signals by activin A do not affect anticancer drug-sensitivity and acquired multi-drug-resistance in human ovarian endometrioid adenocarcinoma OVK-18 cells

Using the human ovarian adenocarcinoma cell line, OVK-18, which is sensitive to activin A-mediated inhibition of growth and various anticancer drugs, we determined whether activin A altered the sensitivity of these cells to seven anticancer drugs. The relationship between the sensitivity to activin and the resistance to anticancer drugs was also investigated in OVK-18 parent cells and OVK-18-derived CDDP-resistant cells. Activin A inhibited proliferation of OVK-18 parent cells in a dose-dependent manner, although it did not affect the sensitivity of OVK-18 parent cells to the seven anticancer drugs, CDDP, CBDCA, adriamycin, paclitaxel, SN38, terarubicin and etoposide (VP16). Both the sensitivity to activin A-mediated inhibition of growth and the sensitivity to anticancer drug-induced apoptosis were reduced in CDDP-resistant cells, while their sensitivity to the seven anticancer drugs was not affected by activin A. Flow cytometric analysis revealed a significant reduction in type IIA activin receptor expression on the surface of CDDP-resistant cells. These results indicate that the activin A-induced intracellular signals inhibiting cell growth are independent of the inhibition caused by the seven anticancer drugs, and suggest that the reduced sensitivity of CDDP-resistant cells to activin A is derived in part from reduced activin receptor expression and not acquired drug-resistance.     

Death-associated protein kinase localization to human renal tubule cells, and increased expression of chronic obstructive uropathy in rats

BACKGROUND: Death-associated protein kinase (DAP kinase) is a Ca2+/calmodulin-dependent serine/threonine kinase implicated as a positive apoptosis mediator. However, little is known about DAP kinase involvement with apoptosis in renal diseases. METHODS: In order to determine whether DAP kinase has a role in renal cell apoptosis in kidney diseases, we performed an immunohistochemical study using a monoclonal antibody to DAP kinase. Firstly, by examining the cellular distribution of DAP kinase in normal human renal tissues and cultured proximal tubule cells. We then used western blotting and immunohistochemical analysis to examine directly whether DAP kinase protein levels could be modulated in rat kidneys with chronic obstructive uropathy created by unilateral ureteric ligation. RESULTS: Immunohistochemistry of normal human kidney tissues showed that DAP kinase was exclusively localized in the cytoplasm of renal tubule cells. Expression analysis of DAP kinase using cultured cells confirmed DAP kinase mRNA and protein presence in human renal tubule cells. Immunocytochemical analysis directly visualized DAP kinase in the cytoplasm of the renal tubule cells in culture. Finally, DAP kinase was found up-regulated in renal tubule cells of rat kidneys with chronic obstructive uropathy. CONCLUSIONS: Our study demonstrates that DAP kinase is localized to renal tubule cells, implying a crucial role for DAP kinase in renal tubular cell apoptosis in progressive renal diseases.     

Strain gauge force transducer and its application in a pig model to evaluate the effect of probiotic on colonic motility

The aim of this study was to investigate the effect of probiotic, i.e., fermented milk prepared with Lactobacillus casei strain Shirota, on colonic motility by the strain gauge force transducer (SGFT) in a pig model. The contractions of the circular muscle layer of the cecum, upper colon, lower colon, and terminal colon in pigs were directly measured in conscious status by this method. This method was useful for quantitatively evaluating the effects of stimuli on colonic motility. Feeding significantly stimulated the motilities of the upper and lower colon. Defecation significantly stimulated the motilities of the upper and terminal colon. Two weeks' feeding of the fermented milk significantly activated the response to feeding in four portions of the large intestine. It increased motility of the terminal colon that did not promote defecation. The frequency of defecation from 9:00 to 10:00 (the period just after the morning meal) increased significantly, but from 0:00 to 1:00 (the midnight period) it decreased as a result of the ingestion of fermented milk. Such effects of the fermented milk on motility of the terminal colon are discussed in relation to the movement of digesta. The effects may relate to the stimulation of colonic fermentation as shown by a decrease in fecal pH.     

依替膦酸二钠对卵巢切除妇女骨密度降低的预防作用

目的 探讨双膦酸盐对切除双侧卵巢的妇女骨密度减低的预防作用。方法 18例早期恶性肿瘤行双侧卵巢切除的患者使用骨吸收抑制剂依替膦酸二钠,400 mg/d×14d,每3个月间断用药一次,持续1年,并以20例因其它病因而实施去势手术的妇女作为对照,研究骨吸收抑制剂依替膦酸二钠对早期恶性肿瘤去势术后妇女骨量的保护作用。结果使用依替膦酸二钠1年时,患者的骨量虽然未能保持在术前水平,但可以显著减少骨量的丢失,与对照组相比差异有显著性(P<0.05),并且骨密度的变化与BALP呈负相关性(P<0.05)。投药1年时肿瘤患者无1例复发。结论依替膦酸二钠可以减少去势手术妇女的骨量丢失,对预防去势手术妇女的骨质疏松有积极的作用。     

The role of p16-cyclin d/CDK-pRb pathway in the tumorigenesis of endometrioid-type endometrial carcinoma

We analysed p16 gene alteration and p16, cyclin-dependent kinase 4 (CDK4), CDK6, cyclin D1, cyclin D2, cyclin D3 and retinoblastoma protein (pRb) expression in ten normal endometriums (PE), 18 endometrial hyperplasias (EH) and 35 endometrial cancers (EC). Two of ten PE (20%), nine of 18 EH (50.0%) and 29 of 35 EC (82.9%) exhibited p16 nuclear staining. p16 expression was significantly higher in EC than EH (P = 0.0119). In the six p16 (-) EC, one was considered to have reduced gene dosage consistent with possible homozygous deletion of the CDKN2 gene and three had methylation in 5'CpG island in the promoter region of the p16 gene, whereas none showed such reduced gene dosage and four had methylation in the nine p16 (-) EH. Strong CDK4 staining was observed in 12 of 35 EC (34.3%) and one of 18 EH (5.6%). The strong expression of CDK4 was higher in EC than in EH (P = 0.0399). The expression of CDK4 was higher in EH than PE (P = 0.0054). The abnormalities of p16-cyclin D/CDK-pRb pathway were detected in 18 of 35 EC (51.4%). In conclusion, the expression of p16 and CDK4 may be an early event in the neoplastic transformation of endometrial cancer.     

Cellular heterogeneity in long-term surviving cells isolated from eutopic endometrial,ovarian endometrioma and adenomyosis tissues

Human endometrial tissues regenerate easily after menstruation and childbirth, suggesting the existence of endometrial stem-like cells that can survive and proliferate from a single cell over a long time. To clarify this hypothesis, limiting dilution cultures performed with eutopic endometrial, ovarian endometrioma and adenomyosis cells obtained from a patient, achieved cloning efficiencies of 13.0, 5.0, and 0.8%, respectively. These monoclonal cells survived for more than 24 months. More than 4 types of monoclonal cells were established from eutopic endometrial cells and microscopically were distinctly different from each other. Intraperitoneal injections of dispersed human eutopic endometrial cells did not cause any endometriosis-like lesions in scid mice, but those of endometrial tissue fragments did. IL-6, TNF-alpha, IL-1beta, M-CSF and HGF failed to enhance transplantation of dispersed endometrial cells to the mice. These results indicate that several types of eutopic endometrial cells survive long-term, and that simple regurgitation of eutopic endometrial stem-like cells may not induce peritoneal endometriosis.