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A research background of pre-eclampsia (pre-eclampsia) is a pregnancy-specific diseases, is a multi-system dysfunction and high blood pressure, urine protein levels, edema main clinical features of the disease, leading to maternal and one of the main reasons for increased perinatal mortality. According to the blood pressure and the degree of proteinuria, pre-eclampsia can be mild pre-eclampsia and severe pre-eclampsia, severe pre-eclampsia is a serious threat to the health of the mother and child. American scholars in the 1980s, the concept of early onset severe preeclampsia (Early-onset severe Preeclampsia, EOSP), Chinese scholar Yang Niyaz onset severe preeclampsia before 32 weeks' gestation is counted as the early onset of the column, After the onset known as late-onset (Late-onset severe Preeclampsia, LOSP). The In recent years, EOSP has caused obstetric Perinatal attention of the medical profession and research interests, EOSP pregnant women the incidence rate of 5% to 7%, accounting for severe preeclampsia 20.4%, due to disease onset gestational weeks early, progression quickly is an important cause of maternal and fetal death, and how to prevent and treat EOSP health workers face a thorny issue. The current research on the pathogenesis of EOSP with LOSP controversial. Kadyrov observed placental implantation site morphology of uterine layer specimens found EOSP patients with placental tissue significantly increased apoptosis, trophoblastic invasion of restricted placental implantation shallow spiral arteries stenosis. Crispi that EOSP prefer a placental disease LOSP with maternal relationship closer. Since most patients with preeclampsia placenta after delivery of clinical symptoms ease soon, so the placental pathology severe preeclampsia relationship increasingly attracted attention. The placenta is an intermediary between maternal and fetal sex organs, placental function is normal or not directly affect the safety of the fetus, scholars placenta as specialized topics to study. With the development of science and technology, the placenta research has been widely involved in the field of Histology and Embryology, Biochemistry, reproductive immunology and molecular biology, but the most basic placental pathology research. In recent years, immunohistochemistry, stereology Ultrasonics widely used in the study of placental pathology the various pathological pregnancy the placenta organizational structure, ultrastructural changes, basically can be observed and detected, and thus clinical diagnosis and treatment play a guiding role. The this stage severe preeclampsia and placental tissue, especially EOSP is mostly morphological studies, relatively few studies of quantitative change. The stereological is a scientific inference by the two-dimensional structural information quantitative three-dimensional structural information. Stereology severe preeclampsia placenta can reflect quantitative changes in the characteristics of severe preeclampsia and placental tissue. Second, the purpose of this article by morphological observation and stereological parameters of severe preeclampsia and placental tissue light microscope level and electron microscope level measurement, to investigate severe preeclampsia placental tissue structures quantitative change, and thus explain the morphological changes characteristic with The relationship between fetal adverse prognosis. Materials and methods. Material: (1) the material source of placental tissue samples taken from Zhujiang Hospital the (refreshing Hospital, Huangqi Hospital) Obstetrics and Gynecology 2007.9 to 2008.6 maternal hospital delivery 40 cases are obtained informed consent of patients. (2) grouping severe preeclampsia group: diagnostic criteria for severe preeclampsia in accordance with the 7th edition of \Of these 20 samples were divided into two groups according to the onset of gestational age: ≥ 32 weeks for late-onset severe preeclampsia group (LOSP group), a total of 10 cases; <32 weeks early onset severe preeclampsia the group (EOSP group), a total of 10 cases. Control group: Select the same period in hospital delivery in preterm and full-term deliveries of 20 cases of placenta as a control group. Of these 20 samples were divided into two groups in accordance with the gestational age: ≥ 32 weeks for the control group A, a total of 10 cases; <32 weeks for the control group B, a total of 10 cases, respectively, as the LOSP the group and EOSP of group control group. The above the LOSP group and control group A, EOSP group and control group B between age, gestational age, producing inferior were not statistically different (P> 0.05). 2 light microscope specimen processing and stereological parameter test set: in the middle of the placenta immediately after delivery of the placenta with, respectively, throughout the five randomly cut size is about 1cm × 1cm x 1cm tissue blocks, fixed in 4% formaldehyde, paraffin package buried randomly selected five sections, HE staining. Each slice 40X objective random shooting of five non-overlapping photos, photographs entered into the computer, and then use the Image-Pro image analysis software design square mesh test system for testing, square mesh point spacing of 30 pixels . Before the test, the first objective micrometer on the grid calibration, calibration the rearward network dot pitch of 9.09 microns. The stereological parameters chosen: the villous capillary volume density (Vv), surface area density (Sv), number density (Nv). Hit the square network test system test the villous capillary and inclusive space of test points, count the number of intersections of the test line with villous capillary, counting the number of villous capillary test area. Vv, Sv, and Nv density parameter reference space for fluff, the number density of the test to the villous capillary wall as a reference. Literature stereological formula to calculate the above parameters. Electron microscope specimen handling and testing: the placenta after delivery, in the middle of the floor area of ??the placenta maternal side immediately with dense fluff at different parts randomly taken from the two organizations, about the size of 2 mm × 2mm x 2mm into saline cleaning wash the blood after immersion in 2.5% glutaraldehyde solution, stored in the refrigerator at 4 ℃. After after fixation, dehydration, immersion, embedded semi-thin locate and repair block ultrathin sections, electron staining, tissue blocks were randomly selected five ultrathin sections, Hitachi H-7500 transmission electron microscopy villi and syncytiotrophoblast cells super micro-structure changes. Shooting the photos do not overlap each slice 15,000 times 5 requires random shooting, shooting accompanied ruler. Shooting electron micrograph entered into the computer, using Image-Pro image analysis software design party network test system for testing, square mesh point spacing of 20 pixels. Before the test, the first electron microscope photo ruler on the grid calibration, calibration the rearward network point spacing was 0.1709 microns. Test as a frame of reference to syncytiotrophoblast, hit the square network test system test mitochondria, endoplasmic reticulum, of microvilli and inclusive space of test points, count the test line with mitochondria, endoplasmic reticulum, the number of intersections of the microvilli, respectively. count test area within the mitochondria, endoplasmic reticulum, and the number of microvilli. Each individual as the parameters to be tested according to the formula. 4. Analysis method: observation the LOSP group and control group A, placental tissue between the EOSP group and control group B microscopic morphology change the stereological parameters between the differences. Data processing and statistical analyzes were performed using the SPSS13.0 statistical package processing data: All data are expressed as mean ± standard deviation ((?) ± SD) said the two sample independent samples t-test, two-sample rate comparison with x ~ 2 test (Fisher's exact, n <40), α = 0.05 significant level of inspection. Results. Placental villi light microscope observations: observation, see A placental villi of the control group has more syncytial nodules rare fibrinoid necrosis fluff with vascular fit membrane (vasculo-syncytialmembrane, VSM) fluff a few more; rare syncytial nodules in the control group B placental villi, fibrinoid necrosis of small, large number of vascular fit membrane fluff; LOSP group the placenta with more syncytial nodules, fibrinoid necrosis than more with the cell membrane of vascular fit fluff few; groups EOSP placenta visible syncytial nodules, fibrinoid necrosis more, while with vascular fit the cell membrane of the villi few. Severe preeclampsia group of terminal villous capillary Vv, Nv were less than the corresponding control group, the difference was statistically significant (p <0.05), Sv, respectively, between the two groups showed no significant difference (P> 0.05) . Placental tissue electron microscopy results: control group A and control group B placenta fit trophoblastic surface have a large number of microvilli, was thin and long, neatly arranged, rich in the endoplasmic reticulum and mitochondria within the cells, the cytoplasm small amount of lipid particles; the the LOSP placental syncytiotrophoblast cell surface microvilli are scarce, the end of the microvilli enlarged part of the syncytiotrophoblast microvilli missing, swelling of mitochondria, endoplasmic reticulum significant expansion of the partial fusion cystic lipid particles increased deposition; EOSP combination trophoblastic except LOSP group's performance, also appeared cytoplasmic vacuolar degeneration, lipid particles increased significantly. Placental syncytiotrophoblast mitochondria stereological parameters: LOSP group with EOSP group placenta syncytiotrophoblast mitochondria Sv, Vv are greater than the corresponding control group, the two groups were compared, the difference was statistically significant (P <0.05); mitochondria the of Nv, EOSP group and LOSP group compared with the corresponding control group, the difference was not statistically significant (P> 0.05). The the placenta syncytiotrophoblast stereological parameters of the endoplasmic reticulum comparison: the LOSP the group and EOSP group placental syncytiotrophoblast endoplasmic reticulum Sv, Vv are greater than the corresponding control group, the two groups were compared, the difference was statistically significant (P <0.05 ); endoplasmic reticulum the of Nv, EOSP group and LOSP group compared with the corresponding control group, the difference was not statistically significant (P> 0.05). Placenta microvilli stereological parameters: LOSP group and EOSP of group placenta syncytiotrophoblast microvilli Sv, Vv, Nv are smaller than the corresponding control group, the two groups were compared, the difference was statistically significant (P <0.05). 3 children perinatal outcome: LOSP cesarean end of pregnancy, 10 cases of fetal growth restriction (fetalgrowth restriction, FGR) 2, neonatal asphyxia two cases, no neonatal deaths; end of EOSP group cesarean section 10 cases of pregnancy , FGR 3 cases, 4 cases of neonatal asphyxia, neonatal death two cases. The end of the control group A cesarean section two cases of pregnancy, natural childbirth eight cases, FGR neonatal asphyxia and neonatal death; control group B 10 cases of natural childbirth FGR0 cases, neonatal asphyxia, neonatal deaths, the two cases. V. Conclusion 1. Severe preeclampsia group observed more adverse neonatal outcomes. 2 light microscope the LOSP the group and EOSP of placental villi presence of ischemia, hypoxia and pathological findings. 3 electron microscope the LOSP the group and EOSP of group the placenta syncytiotrophoblast cell ultrastructure serious damage. 4. In severe preeclampsia environment, placental villi and fit trophoblast cell ultrastructure, caused by the change of its physiological functions, affecting transport and synthesis functions of the placenta may be involved in fetal injury.
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