• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.305-312
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• 2007

Objective: To evaluate the efficacy of split insemination method in treatments for non-male factor infertility. Method: Laboratory and clinical data were collected from 505 cycles of split insemination during 2002$\sim$2005 in our center. The subjects were non-male factor infertility such as endometriosis, tubal, uterine, PCOS and idiopathic infertility without any sperm defects. Retrieved oocytes were randomly divided, and inseminated by conventional IVF or ICSI. Fertilized zygotes were cultured for 2$\sim$5 days to ET date, and surplus zygotes and embryos were frozen for subsequent frozen-thawed ET cycles. Clinical outcomes according to insemination method were compared by statistical analysis. Results: The overall fertilization per retrieved oocytes was significantly higher in ICSI than that of conventional IVF in sibling oocytes (62.5$\pm$22.3% vs 52.9$\pm$28.0%, p<0.01). Total fertilization failure occurred only in 2 of 505 cycles (0.4%) in split insemination cycles. Incidence of fertilization failure and poor fertilization rate less than 30% by ICSI were significantly lower than those of conventional IVF (1.1% and 7.5% vs 8.5% and 22.0%, p<0.01). Delivery rates after transfer of fresh and thawed embryos from split insemination cycles were 40.0% (185/462) and 35.0% (55/157), respectively. There was no significant difference in the implantation and delivery rates of ET with embryos from conventional IVF or ICSI. Conclusion: Taken together, the split insemination method improves poor fertilization rates resulting in successful clinical outcomes and thus could be used for non-male factor infertile couples in human ART program.

• Childhood Kidney Diseases
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• v.10 no.1
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• pp.8-17
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• 2006

Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.