Objective To explore the effect of hydrogen peroxide (H2O2) on the expression of human leukocyte antigen-G ( HLA-G) in placental trophoblasts in pregnant women with pre-eclampsia.Methods Forty pregnant women,delivered through cesarean section in the Department of Obstetrics of and Gynecology,the First Affiliated Hospital of Nanjing Medical University from October 2008 to October 2009,were enrolled,including 20 women with pre-eclampsia and 20 healthy gravidas (control group).Colorimetry and western blot were applied,respectively,to determine the level of H2O2 and the expression of HLA-G protein in placental tissues and the correlation between them were analyzed.After 24 hours of seeding,JEG-3 cells (the HLA-G positive cell line of choriocarcinoma) were divided into two groups:intervention group (exposure to 175 μmol/L H2O2) and control group (without H2O2).Immunofluorescence and western blot were used to investigate the expression of HLA-G protein in JEG-3 cells at 24 hours and 48 hours after incubation.Results (1 )The level of H2O2 in placenta in the pre-eclampsia group was significantly higher than that in control group[(105 ±13) nmol·mg-1·prot-1 vs (62 ± 18) nmol·mg-1 ·prot-1,P < 0.05].(2) The expression of HLA-G protein in placenta of the pre-eclampsia group was reduced by88%compared with that of the control (0.20 ± 0.08 vs 1.67 ± 0.65,P < 0.05).( 3) Negative correlation was found between HLA-G level and H2O2 expression in the placenta in both groups(r =-0.895,P =0.000).(4) Compared with the control group,the expression of HLA-G protein in JEG-3 cells,after 24 hours and 48 hours exposure to H2O2,reduced by 39% and 80%,respectively,(3.21 ±0.33 vs 1.95 ±0.25 and 0.65 ±0.08,P <0.05,respectively) and 67% reduction was detected from 24 hours to 48 hours of H2O2 exposure (P <0.05).Fluorescence microscope observed reduced expression of HLA-G in JEG-E cells in the intervention group at 48 hours compared to the control group (P<0.05).Conclusion High level of H2O2 could down-regulate HLA-G expression in the placental trophoblasts in pre-eclampsia which may be involved in the pathogenesis of pre-eclampsia.

Objective:To study the expression of aromatasecy P450 and receptor of estrogen and progesterone in the tissue of endometrial hyperplasia and endometrial cancinoma,analysis the correlation of the endometrial carcinoma pathogenesis,clinical stage and cell differentiation.Methods:The immunohistochemiscal S-P method techniques were performed to study the expression of aromatasecy P450,estrogen receptor(ER) and progesterone receptor(PR) in the tissue of endometrium from 226 cases,including 42 cases of normal endometrium,52 cases of simple endometrial hyperplasia,38 cases of complex endometrial hyperplasia,36 cases of endometrial atypical hyperplasia and 58 cases of endometrial carcinoma.Results:There was no the positive expression in the normal endometrial tissues.But the positive expressions of aromatasecy P450 were found in the tissue of endometrial hyperplasia(simple endometrial hyperplasia,complex endometrial,atypical hyperplasia)and endometrial carcinoma.The rates of the positive expression were 40.4%,47.4%,69.4%,72.4% respectively.There were significant relationships among fifth groups(P

Objective To evaluate the relationship between pathogenesis of preeclampsia (PE) and the ultrastructure change of the endoplasmic reticulum in trophocyte, mRNA and protein expression levels of endoplasmic reticulum molecular chaperone glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94), endoplasmic reticulum apoptosis factor cysteine protease protein 12 (caspase-12).Methods Sixty-five pregnant women who were hospitalized in the First Affiliated Hospital of Nanjing Medical University from July 2008 to January 2010, were selected as the subject. Thirty pregnancy women diagnosed with PE were divided into PE group and 35 normal pregnant women were used as control group.Electron Microscopy was used to measure ultrastructure change of the endoplasmic reticulum in placenta trophocyte. Reverse transcription(RT) PCR and western blot were used to investigute the expression levels of GRP78, GRP94, caspase-12 mRNA and protein in placenta. Results (1) In control group the volume of endoplasmic reticulum does not increase; no swelling and no expansion of endoplasmic reticulum was found.In PE group the edema number of endoplasmic reticulum was reduced; the volume of endoplasmic reticulum increased; expansion and vacuolation of cavity and degranulation of the endoplasmic reticulum was observed significantly. (2) The mRNA and protein expression levels of GRP78 in placenta of PE group (2.59 ± 0. 09 and 0. 81 ±0. 31) were significantly higher than those in placenta of control group (1. 16 ±0. 07 and 0. 40 ± 0. 10, P <0. 01). (3) The mRNA and protein expression levels of GRP94 in placenta of PE group (1.31 ± 0. 91 and 0. 55 ±0. 24) were significantly higher than those in placenta of control group (0. 63 ±0. 57 and 0. 22 ±0. 09, P < 0. 01). (4) The mRNA and protein expression levels of caspase-12 in placenta of PE group (4. 03 ± 0. 65 and 1.56 ± 0. 17) were significantly higher than those in placenta of control group (1.85 ± 0. 85 and 0. 91 ± 0. 69, P < 0. 01). Conclusion The obvious expansion of endoplasmic reticulum in trophocyte and the increased expression levels of GRP78, GRP94 and caspase-12 indicate that endoplasmic reticulum stress-mediated apoptosis may be involved in the pathophysiological processes of PE.

Objective To investigate the effect of epidermal growth factor (EGF) on the expression of Matrix metalloproteinase-9 (MMP-9) and the signalling pathways involved in the trophoblast cell line JEG-3. Methods The JEG-3 trophoblast cell line was used in this study. (1) JEG-3 cells were cultured with various concentrations of EGF (0, 1, 10,20 ng/ml) for 24 hours and the expression of MMP-9 was tested by western blotting and reverse transcription PCR (RT-PCR). (2) Western blotting and RT-PCR were also used to investigate the expression of MMP-9 expression after incubation for 0,4,12 and 24 hours with EGF treatment (10 ng/ml) in JEG-3 cells. (3) According to the different added ingredients, JEG-3 cells were divided into some groups: control group (without EGF), EGF group (exposure to l0ng/ml EGF),EGF + inhibitors group (exposure to 10 ng/ml EGF +20 ng/ml SB203580 or exposure to 10 ng/ml EGF + 10ng/ml U0126) inhibitors group (exposure to 20 ng/ml SB203580 or exposure to 10 ng/ml U0126). Western blotting were used to investigate the expression levels of MMP-9, nuclear factor kappa B (NF-kB) ,p38MAPK, phospho-p38MAPK (p-p38MAPK) , extracellular -signal regulated kinase (ERK) and phospho-ERK (p-ERK) protein in JEG-3 cells after incubation for 24 hours. Results (1) The profiles of MMP-9mRNA were increased by various concentrations of EGF (0, 1 , 10, 20 ng/ml) in JEG-3 cells after 24hculture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0. 567 ±0. 056) , 10ng/ml of EGF (1. 392 ±0. 133) , 20 ng/ml of EGF (1. 971 ±0. 067) were significantly higher respectively (P <0. 05) , compared with 0 ng/ml of EGF treatment (0. 166 ±0. 015). Similarly, MMP-9 mRNAs were also increased with the increasing incubation time. Compared to EGF (10 ng/ml) stimulation for 0 h (0.253 ±0.044), the MMP-9 mRNA profiles were 0. 470 ±0. 026, 1.061 ±0. 115, 1. 453 ±0. 180 for 4,12 and 24 hours, respectively (P < 0. 05). (2) In accordance to the mRNA profiles, the expression of MMP-9 protein was also increased by different concentrations of EGF (0,1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The abundance of MMP-9 protein in the three groups was 0. 043 ±0. 012, 0. 085 ±0. 008, 0. 142 ±0. 015, with a significantly higher expression, compared with 0 ng/ml of EGF treatment (0. 004 ±0.001, P < 0.05) respectively. Similarly, MMP-9 proteins were also increased with the increasing incubation time. Compared to EGF(10 ng/ml) stimulation for 0 h (0. 030 ±0. 009) , the profiles of MMP-9 protein were 0. 137 ± 0. 010, 0. 240 ± 0. 010, 1.240 ±0.061 for 4, 12 and 24 hours, respectively (P < 0.05). (3) Both p38MAPK and ERK signalling pathways were activated by EGF in JEG-3 cells. The expression of p-p38MAPK was significantly higher (without or with 10 ng/ml EGF, 234. 1 ± 4. 1 vs.260. 9 ± 2. 5 , P < 0. 05) , however, the p38 MAPK inhibitor SB203580 markedly suppressed the increase in p-p38MAPK content induced by EGF(227. 9 ±2. 4 vs. 260. 9 ±2. 5, P<0. 05). Similarly, the expression of p-ERK was significantly higher with EGF treatment (812. 2 ±3. 5) vs. without EGF group (453.4±5.8) (P <0. 05) , while the ERK inhibitor U0126 significantly inhibited the increased p-ERK content in response to EGF treatment (71. 0 ± 1. 2 vs. 812. 2 ± 3. 5, P < 0. 05) . (4) The p38MAPK inhibitor SB203580 significantly reduced the expression of EGF-induced MMP-9 (0. 645 ± 0. 270 vs. 1. 476 ± 0. 452, P < 0. 05)and NF-kB (0.530 ± 0.026 vs. 0.959 ± 0. 017, P < 0. 05) . (5) The ERK inhibitor U0126 also significantly reduced the expression of EGF-induced MMP-9 (0. 623 ±0. 030 vs. 2. 112 ±0. 056, P <0. 05)and NF-kB (0. 325 ± 0. 082 vs. 0. 939 ± 0. 153, P < 0. 05). Conclusion EGF induced the expression of MMP-9 in a time and dose-dependant manner in JEG-3 cells. EGF enhanced MMP-9 expression through the activation of p38MAPK and ERK signalling pathways in JEG-3 cells.

Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.

Objective To explore the function of placental trophoblast cell apoptosis on the pathogenetic mechanism of pregnancy induced hypertension (PIH). Methods Apoptosis of trophoblast cells in 20 cases of PIH(PIH group) and in 10 cases of normal pregnancy (control group) were directly observed using the terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL) method. Apoptosis gene expression patterns were screened with gene chip provided by Poxing Company, Shanghai. Standards for differently expressed genes were: (1) An absolute value of the natural logarithm of cy5(PIH group)/cy3(control group) greater than 0.69 with a difference of signal of cy5 2 times over that of cy3. (2) The signal value either cy3 or cy5 must be greater than 800. Results (1) TUNEL test showed that the number of trophoblast cells apoptosis per ten thousand ?m 2 was 1.584 in the PIH group and 0.032 in the control group with significant difference between the two groups (P

PurposeTraumatic pancreatitis which has a high mortality rate is likely to be misdiagnosed. This study aims to analyze the clinical manifestations and CT findings of traumatic pancreatitis, so as to improve its early diagnosis and treatment.Materials and Methods The clinical manifestations and CT images of 25 patients with traumatic pancreatitis confirmed by operation or post-treatment review were analyzed retrospectively. Pancreatic injuries were classified as superficial lesions (with the depth of trauma less than 50% of the thickness of pancreas) and deep lesions (with the depth of trauma more than 50% of the thickness of pancreas). The clinical manifestations, CT findings and the complicated organ injuries in these two types of pancreatic trauma were analyzed.Results Eight patients had superficial lesions, and 17 patients were with deep lesions. Nine patients had complicated organ injuries. Patients with deep lesions showed a more severe abdominal pain, nausea, vomiting, rebound tenderness and muscular tension than those patients with superficial lesions. The serum amylases increased in all the patients. Pancreatic-relevant complications including pancreas pseudocyst, pancreatic fluid leakage and peritonitis occurred in 7 patients who accepted a delayed operation. Three out of 8 patients with superficial pancreatic injuries were missed on plain CT scan in the first time. Among 17 patients with deep pancreatic trauma, 12 had incomplete laceration, 5 had complete laceration, and 1 was missed in the first time. The direct CT features of pancreatic trauma were focal abnormal attenuation and/or discontinuity in pancreatic parenchyma.Conclusion The clinical manifestations of patients with traumatic pancreatitis are complicated. The direct CT features of pancreatic trauma include heterogeneous density of pancreatic parenchyma and/or interruption. Trauma's depth is closely related to the main injury of pancreatic duct. It is worth to be aware of the indirect signs such as peripancreatic oozy and other viscera damages.

Objective To analyze reasons for unplanned return-to-theater obstetrical surgery in patients with placenta previa, and to propose a strategy for prevention.Methods Among 571 patients with placenta previa in the Department of Obstetrics, First Affiliated Hospital of Nanjing Medical University from January 2010 to January 2015, ten cases (1.75％) who had an unplanned return-to-theater obstetrical surgery were retrospectively analyzed.Results Seven out of the ten cases returned to the theater due to severe hemorrhage after cesarean section and hysterectomy or uterine artery embolization was performed.The rest three pregnancies were terminated at mid-term with amniotic injection of rivanol, two of which developed severe infection after the induction combined with uterine artery embolization followed by cesarean section,and the other one finally had an emergent hysterectomy due to severe postpartum hemorrhage after cesarean section because of intrapartum hemorrhage.Severe postpartum hemorrhage occurred in eight out of the ten cases, with a mean volume of (4 212± 1 651) ml.Blood loss between the original and return-to-theater surgery was (2 206± 736) ml.In these eight cases, the mean volume of erythrocyte suspension transfusion was (23.7±9.0) U, and [M(min-max)] 1 845(390 3 960) ml for plasma transfusion.Platelet transfusion was performed in five cases, cryoprecipitate transfusion in eight cases, serum albumin transfusion in six cases, and fibrinogen transfusion in five cases.The interval between original and return-to-theater surgery was 2.0(0.5-19.0) h.After the return-to theater surgery, the time of antibiotic use was (9.2±2.3) d, and the duration of hospital stays was (10.6±2.5) d.No patient required further re-operation, and all were discharged without long-term sequelae.All seven neonates had a good prognosis.Conclusions Severe postpartum hemorrhage in patients after initial operation because of placenta previa is the primary indication for unplanned return-totheater surgery.Closed postoperative monitoring, early recognition and expedite return-to-theater surgery are crucial to stop bleeding and save lifes.

Objective Present study aimed to characterize the alteration of cortical thickness in first-episode, never-medicated, adult patients with major depressive disorder (MDD), and explore whether such deficits were related with their disease duration and clinical symptom severity. Methods Thirty-seven adult MDD patients were recruited from March 2013 to August 2015 as patient group, and 41 healthy volunteers were as control group. All the patients underwent three-dimensional spoiled gradient recalled (3D-SPGR) sequences, and the images were acquired. Constructions of the cortical surface were developed from 3D-SPGR images using FreeSurfer software, and the thickness of the entire cortex was measured according to the automated surface reconstruction, transformation, and high-resolution inter-subject alignment procedures. Finally, cortical thickness was compared between the two groups, and the relativity between clinical symptom severity, disease progression and clinical scores were analyzed using the General Linear Model (GLM). Results Our results revealed a significant increase in cortical thickness(P<0.05, false discovery rate corrected) in the left anterior and middle cingulate cortex, bilateral precentral cortex, left paracentral cortex, bilateral superior parietal cortex, left temporal pole, and right lateral occipital cortex (cortical thickness 1.89-2.87 mm, cortical volume 34-384 mm2, P<0.05) in MDD patients compared to healthy controls, while no reversed alternation was found. In addition, clinical symptom severity and disease progression showed no correlation with the cortical thickness abnormalities in MDD group(P>0.05). Conclusion Excluding the impact of treatment, our study showed that the cortical thickness change was mainly located in the prefrontal-limbic system in the in early course of MDD.

Objective To find out the bacterial species in the vagina of postpartum women and the possible influencing factors on colonization. Methods From Jun. 2007 to Oct. 2007, 560 postparmm women from 7 hospitals in China were enrolled. Questionnaire survey, gynecological examination and Nngent score of vaginal smear and microbial spectrum study of the vaginal flora were completed. Results (1 ) According to the Nngent score, 48 out of the 560 women were normal (8.6%), 337 at the borderline (60. 2% ) and 175 (31.2%) were complicated with bacterial vaginosis (BV). Among the 560 women, Bacterium lacticum were identified in 74 cases (13. 2% ), but not in the rest 486 cases (86. 8% ). Gardnerella and bacteroids were detected in 322 women (57. 5% ) and small flectobacillus in 214 women (38. 2% ) out of the 560 subjects. (2) Influencing factors on vaginal microflora: among the 266 women who had normal vaginal delivery, 23 (9. 4% ) showed normal vaginal microflora, 148 (55.6% ) at borderline and BV was diagnosed in 93 women (35.0%). The corresponding figures among the 294 women who underwent cesarean section were 23 (7. 8% ), 189 (64. 3% ) and 82(27.9% ), respectively. However, the incidence of BV had no statistical difference between these two groups (P = 0. 204). In the 233 women who received episiotomy, 22 (9. 4% ) showed normal vaginal microflora, 135 (57.9%) at borderline and 76 presented with BV (32. 6% ), the corresponding figures among the 327 women without episiotomy were 26 (8. 0% ), 202 (61.8%) and 99 (30. 2% ), respectively. The incidence of BV did not show any statistical difference between the above two groups (P = 0. 790 ). (3 ) Prenatal vaginitis were reported in 46 women, among which 5 (10.9%) with normal vaginal flora, 26 (56.5%) at borderline and 15 (32.6%) with BV, and again in the 514 women without prenatal vaginits, the above figures changed to 43 (8.4%) , 311 (60. 5% ) and 160 (31.1%). No significant difference was found in the incidence of BV between the two groups (P =0. 962). The rate of BV in women without sex, with sex occasionally and with sex frequently during pregnancy was 27.5% (78/284), 35.6% (96/270) and 1/6, respectively (P = O. 185), and the numbers in women who had breast-feeding, bottle feeding and mixed feeding were 31.0% (67/216), 39.3% (35/89) and 28.6% (73/255), respectively (P=0.573). Conclusions The amount of Lactobacillus in vagina of postpartum women is greatly reduced leading to dysbacteria. The incidence of BV is not affected by vaginal delivery, episiotomy, vaginitis, prenatal intercourse and the way of feeding, but is higher in postpartum women.