Objective To investigate the role of transcription factor Ets-1 involved in premature rupture of membranes(PROM) by detecting its expression and location in human fetal membranes. Methods Between Feb. and Nov. 2007, 100 pregnant women who delivered in the First Affiliated Hospital, Chongqing Medical University were enrolled in this study. According to gestational weeks, rupture of amuiochorionic membranes and delivery mode, those women were classified into preterm labor group, preterm premature rupture of membranes (PPROM) group, term labor group and term premature rupture of membranes (TPROM) group, matched with elective term cesarean sections women as control group. There were 20 pregnant women in every group. The expression and distribution of Ets-1 protein in fetal membranes were detected by immunohistochemical streptravidin-biotin peroxidase(SP) method and histoscore. In the mean time, 6 cases were chosen from each group randomly and reverse transcription-PCR was used to measure the Ets-1 mRNA expression. Results (1) The expression of Ets-1 mRNA in fetal membranes were 0.342±0.016 in preterm labor group,0.603±0.027 in PPROM group,0.325±0.013 in term labor group, 0.582±0.075 in TPROM group,0.139±0.012 in control group, respectively. When compared with that in control group, Ets-1 mRNA expression were significantly increased in both PPROM and TPROM group(P< 0.05). However, it did not show remarkably difference between preterm group and term labor group, as well as between PPROM and TPROM group (P>0.05). (2) Ets-1 was in stromal layers of amniochorionic membranes and in both cytoplasm and nuclei of trophoblast, which were shown with diffuse intracellular positive granularities (brown-yellow) clearly. No expression of Ets-1 was observed in amniotic epithelial cells. (3) The expression of Ets-1 protein was 0.552±0.018 in preterm labor group, 2.853±0.174 in PPROM group, 0.538±0.042 in term labor group, 2.731±0.090 in TPROM group and 0.214±0.013 in control group, respectively. Ets-1 protein was significantly increased in the stroma of both PPROM and TPROM group than that in control group(P<0.05). However, no remarkable different expression of Ets-1 was observed between preterm and term labor group,so was that between PPROM and TPROM group(P> 0.05). Conclusion Transcription factor Ets-1 is expressed in human amniochorionic membranes and it could be up-regulated in PROM.

Objective To evaluate the expression of Gadd45α and p38 MAPK in placentas and the correlations of Gadd45α protein and serum soluble vascular endothelial growth factor receptor-1 (sFlt-1) and soluble endoglin (sEng) in preeclampsia(PE). Methods Fifty-four pregnant women who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the subjects. They were classified into mild preeclampsia group (n=20),severe preeclampsia group (n=16) and the control group (normal pregnant women underwent elective cesarean sections at term without labor and perinatal complications, n=18). Western blot and immunohistochemistry were employed to determine the expression and localization of Gadd45α and p-p38 MAPK protein respectively. Gadd45α mRNA level was determined by quantitative real-time PCR. The levels of seum sFlt-1 and sEng were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and LSD-t test were applied for statistical analysis. Results (1)Immunohistochemistry identified that the positive stained cells were mostly located in trophoblast cells in normotensive placentas, whereas in preeclamptic placentas Gadd45α protein and p-p38 MAPK protein were detected in trophoblast and endothelial cells, as well as a few stromal cells at increased levels.(2)The mRNA levels of Gadd45α was significantly elevated in mild and severe preeclampsia groups compared to the control group (2.10±0.11 and 3.33±0.13 vs 1.01±0.18, P＜0.05), and Gadd45α mRNA level in severe group was significantly higher than in mild group (P＜0.05).(3)The data of Western blot revealed that the Gadd45α protein levels in each group were 0.22±0.11, 0.65±0.15 and 1.34±0.17, respectively, with significant differences between each group(P＜0.05). The p-p38 MAPK protein levels in each group were 0.32±0.08, 0.72±0.12 and 1.45±0.21, respectively, with significant differences between each group (P＜0.05). p38 MAPK protein levels in the total groups showed no difference(P＞0.05).(4)Compared with the control group, sFlt-1 and sEng concentrations in maternal circulation were significantly increasing in mild and severe preeclampsia groups, and concentrations in severe group were significantly higher than those in mild group (P＜0.05).(5) There were positive correlations between Gadd45α protein levels and the concentrations of serum sFlt-1 and sEng in each group( r=0.88 and 0.87, respectively all P＜0.05). Conclusions Upregulation of Gadd45α in preeclampsia placentas may play an important role in the pathogenesis of preeclampsia. It may induce the increased maternal serum levels of sFlt-l and sEng by activating p38 MAPK signaling pathway, leading to deficient cytotrophoblastic invasion and abnormal placental vascular reconstruction during pregnancy.

Anemia, Hemolytic ; complications ; Child ; Humans ; Pneumonia ; complications

Objective To evaluate the expression and location of the growth arrest and DNA damage 45 alpha(Gadd45α)gene in human placenta and explore the relationship between Gadd45α and preeclampsia(PE).Methods Thirty-six women with preeclampsia who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the study objects.They were classified into mild group( n = 20), severe group( n = 16) and elective term cesarean sections group without previous onset of labor and perinatal complications ( control group, n = 18 ).Placentas were collected after they delivered and immunohistochemical streptravidin-biotin peroxidase(SP) method and histoscore were employed to detect the expression and localization of Gadd45α protein.Gadd45α mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR) technique and western blot analysis was used to quantify Gadd45α protein expression level.Results (1)Gadd45α protein was detectable in placenta tissues of the mild and severe groups, mostly located in cytoplasm and nuclei of trophoblast, plus nuclei of vascular endothelial cells and a few stromal cells; whereas placenta tissues of control group showed weak staining, and only detectable in trophoblast, undetectable in vascular endothelial cells.(2)The Gadd45α mRNA levels in placenta tissues of the severe and mild groups were 0.75 ±0.07 and 0.44±0.13, respectively, significantly higher than that in control group (0.18 ±0.04, P ＜0.05);compared with mild group, Gadd45α mRNA level was significantly higher in severe group( P ＜ 0.05 ).(3)The Gadd45α protein levels in placenta tissues of the severe and mild groups was 1.34 ±0.17 and 0.65 ±0.15, respectively, compared with mild group, Gadd45α protein level was higher in severe group (P ＜0.05); they were both significantly higher than that in control group ( 0.22 ± 0.11, P ＜ 0.05 ).Conclusions Gadd45α mRNA and protein levels in placenta tissues of patients with preeclampsia are higher than that of normotensive women, and up-regulated with aggravation of preeclampsia; the Gadd45α protein is located in trophoblast cells, which are closely related to pathogenesis of preeclampsia.These results indicate that Gadd45α plays an important role in the pathogenesis and progression of preeclampsia.

To cultivate more excellent pharmacists, ensuring patients rational drug use, standardiza-tion training of hospital pharmacists in Beijing has been carried out since 2000, and the wonderful effect has been achieved. The teaching methods mainly include centralized training model, small lectures, practice teaching and self-study. The training teaching content mainly includes prescription audit, drug dispensing and management as well as the pharmaceutical care. In this paper, based on the analysis of the status quo, it is recommended that detailed training formulate, teaching methods and content of the second stage of the training about the training management policy, and the clinicians teaching should be added to the contents of teaching. The thought moral qualities, laws and regula tions, as well as academic and research training should also be added to the teaching content.

Objective To explore the expression of pentraxin-3 (PTX3) in placentas from patients with severe preeclampsia and the relationship between PTX3 and the pathogenesis of severe preeclampsia.Methods Fifty-three pregnant women who delivered from October 2010 to March 2011 in the First Affiliated Hospital of Chongqing Medical University were included in the study.Twenty-three women with severe preeclampsia were chosen as the preeclampsia group,and thirty healthy pregnant women were identified as the control group.All the women received cesarean section.The location of PTX3 protein in placentas was studied by immunohistochemical SP method.Quantitative real-time PCR technique and western blot analysis were employed to assay the levels of PTX3 mRNA and protein in placentas,respectively.Results ( 1 ) The location of PTX3 protein in placentas:PTX3 protein was expressed in placentas from both groups,and there was no difference of PTX3 distribution between normal and preeclamptic placentas.PTX3 was mainly located in perivascular stroma,decidual cells and terminal villi.Neutrophilic infiltration was observed in the preeclamptic placentas.(2)The expression of PTX3 mRNA and protein in placentas:the level of PTX3 mRNA in placentas from the preeclampsia group was higher than that in the control group( 1.98 ± 0.54 vs.0.87 ± 0.27,P ＜ 0.05 ).Compared with the control group,the level of PTX3 protein was significantly elevated in the preeclampsia group ( 1.42 ± 0.29 vs.0.56 ± 0.25,P ＜ 0.01 ).Conclusion The high expression of PTX3 in placentas from the preeclamptic patients suggests that PTX3 may be involved in the pathologic process of preeclampsia.

Objective To analyze the clinical characteristics and perinatal outcome of early-onset intrahepatic cholestasis of pregnancy (ICP).Methods A total of 305 ICP cases were collected in the First Affiliated Hospital of Chongqing Medical University between June 2006 and May 2012.According to the onset time of ICP,patients were divided into early-onset ICP group (onset time ＜ 28 gestational weeks) and lateonset ICP group (onset time ≥28 gestational weeks).The late-onset ICP group was further divided into 28-31 +6 gestational weeks and ≥32 gestational weeks according to the onset time.The biochemical indices and perinatal outcome of each group were assessed.Results (1) When the diagnosis was made for the first time,the maternal serum concentrations of total bile acid (TBA) and total bilirubin (TBIL) in early-onset ICP group were (41 ±9) and (32 ±9) μmol/L,respectively; while TBA and TBIL in late-onset ICP group were (32 ± 6) and (22 ± 9) μmol/L,and the difference between the two groups was statistically significant (P ＜ 0.05).(2) There was no significant difference in alanine aminotran-sferase (ALT) and aspartate aminotransferase (AST) between early-onset ICP group and late-onset ICP group (P ＞ 0.05).The ALT of early-onset ICP group and late-onset ICP group were (159 ± 50) and (145 ± 52) U/L,respectively; and AST were (151 ±49) and (138 ± 44) U/L,respectively.(3) The early-onset ICP group had significant higher (P ＜ 0.05) incidence of meconium staining (18.8％ vs.7.4％),fetal distress (22.9％ vs.8.9％),newborn asphyxia (14.6％ vs.5.4％),premature delivery (33.3％ vs.15.6％),developing into severe ICP (41.7％ vs.25.3％) and cesarean section (91.7％ vs.78.6％) when compared to the late-onset ICP group.No significant difference in the incidence of premature delivery,developing into severe ICP and cesarean section was found between the two types of late-onset ICE (4) There was significant differences in average birth weight and gestational weeks at delivery between the two groups [early-onset ICP group:(3113 ± 443) g and (36.3 ± 2.6) weeks] ; late-onset ICP group:[(3513 ± 450) g and (37.7 ±1.6) weeks].Conclusion The early-onset ICP patients presented worse clinical manifestations than lateonset ICP patients,and early-onset ICP is more likely to lead to premature delivery and fetal distress.

Objective To explore the effect of oxidative stress on human Wiskott-Aldrich syndrome related protein 2 (WAVE2) expression in placental trophoblasts in women with preeclampsia.Methods (1) Twenty women with preeclampsia and twenty-three normal term pregnant women,delivered from August 15,2011 to February 23,2012 in the First Affiliated Hospital of Chongqing Medical University,were recruited and divided into preeclampsia group and control group.Placenta samples were collected after cesarean section.The localization and distribution of WAVE2 in placenta was studied by immunohistochemistry.Quantitative real-time polymerase chain reaction and Western blot were employed to assay the WAVE2 mRNA and protein levels.Tissue homogenates was applied to determine the levels of reactive oxygen species (ROS).The correlation between ROS levels and WAVE2 was also analyzed.(2) An in vitro hypoxia/reoxygenation (H/R) model was utilized to simulate ischemia/reperfusion injury to placental trophoblasts.The HTR-8/ SVneo cells (immortalized human first trimester extravillous trophoblast cells) were pre-incubated overnight,after exposure to H/R or normoxic conditions for 48 hours.Flow cytometry was employed to analyze intracellular ROS level.Meanwhile,Transwell assay was utilized to analyze the invasion and migration of HTR-8/SVneo cells.The location and expression of WAVE2 in trophoblasts was evaluated by cell immunofluorescence and Western blot.Statistical differences between the two groups were evaluated by independent t-tests.Pearson's correlation coefficient test was used for correlation analysis.Results (1)Compared with the control group,preeclampsia group had significantly higher 24-hour proteinuria [(1.96±0.24) g vs (0.08±0.05) g,t=19.436,P＜0.05],systolic blood pressure [(154 ± 13) mm Hg vs (98 ±11) mm Hg,t=11.324,P＜0.05] and diastolic blood pressure [(105±14) mm Hgvs (69±8) mm Hg,t=9.612,P＜0.05].In addition,the placental weight and birth weight of infants in preeclampsia group were significantly reduced as compared to the control group [(432±53) g vs (536±67) g,(2446± 187) g vs (3207± 233) g,t=14.562 and 16.307,allP＜0.05)].The WAVE2 mRNA level (0.28±0.07 vs 1.01±0.02,t=12.747,P＜0.05) and the WAVE2 protein levels (0.63±0.08 vs 1.34±0.19,t=11.648,P＜0.05) were also significantly decreased in preeclampsia groups.The level of ROS in placenta in the preeclampsia group was significantly higher than in control group [(144.22 ± 12.32) nmol/(mg · prot) vs (75.17 ± 8.71) nmol/(mg · prot),t=20.467,P＜0.05].There was significant negative correlation between ROS level and WAVE2 protein expression in preeclamptic placenta (r =-0.726,P =0.000).(2) In vitro study showed that,the levels of ROS in normoxia group and H/R group was (82.9±5.8)％ and (155.6±8.1)％,(t=12.747,P＜0.05).Compared with normoxia condition,decreased cell invasion and migration were found in HTR-8/SVneo cells in H/R group [(51.9 ± 3.3)％ and (58.4 ±4.2)％ respectively,t=11.034 and 13.839,P＜0.05].Results from the cell immunofluorescence showed that WAVE2 protein located in the cytoplasm of HTR-8/SVneo cells,and the expression of WAVE2 protein was significant decreased in HTR-8/SVneo cells after exposure to H/R for 48 h (0.37±0.05 vs 0.76±0.06,t=8.631,P＜0.05).Conclusions Excessive oxidative stress in preeclamptic placentas was correlated with the decreased expression of WAVE2.H/R-induced oxidative stress could decrease WAVE2 expression,which may contribute to impaired trophoblast invasion and migration in preeclampsia.

Objective To explore the optimal timing of termination of pregnancy,we analyzed the different gestational age in repeat cesarean delivery and maternal and neonatal outcomes.Methods This was a retrospective study.The information of cesarean sections was collected from maternal obstetric records in the electronic medical recording system of the First Affiliated Hospital of Chongqing Medical University from June 1,2011 to June 30,2013,and women with intrauterine viable singleton pregnancies delivered after 37 weeks of gestation without prenatal complications were selected.They were divided into five groups with different gestational weeks.Maternal general information,perioperative outcome and rate of neonatal adverse event were analyzed with one way ANOVA analysis and Chi-square test.Results A total of 579 cases of elective repeat cesarean at term were performed.The ratios of cesarean section prior to 39 and 39-39+6 weeks of gestation were 64.6％ (374/579) and 29.0％ (168/579),respectively.No fetal,neonatal or maternal death occurred.There were no statistically significant differences in the termination of pregnancy at 37-37+6 weeks,38 38+6 weeks,39-39+6 weeks,40 weeks and ≥ 41 weeks between the two time intervals for cesarean section (P＞0.05).There were statistically significant differences in the length of hospitalization [(4.9±3.0),(4.3 ± 1.3),(4.3 ± 1.0),(4.5± 1.2) and (4.0±0.7) d,respectively; F=2.849,P＜0.05].No significant difference was observed in the maternal BMI,placental membrane residue,maternal perioperative bleeding,premature rupture of membrane (PROM),intensive care unit (ICU) admission and uterine resection (P＞0.05).There were statistically significant differences among the five groups in neonatal weight [(3 082.9±479.2),(3 318.1 ±390.8),(3 415.7±431.1),(3 630.5±475.2) and (3 334.0±242.5) g,F=13.798] and length [(48.8± 1.5),(49.3± 1.5),(49.6± 1.5),(50.0± 1.5) and (47.8±3.9) cm,F=7.460; both P＜0.05].One min and 5 min Apgar scores also showed statistically significant differences [1 min:(9.7±0.7),(9.8±0.6),(9.8±0.4),(9.7±0.5) and (8.8±2.7) ; F=4.432; 5 min:(9.9±0.3),(10.0±0.3),(10.0±0.2),(10.0±0.2) and (9.2± 1.8),F=9.625; all P＜0.05].The overall rates of neonatal adverse events,including the admission to neonatal intensive care units (NICU),the rates of cardiopulmonary resuscitation or ventilator therapy,asphyxiation,as well as the length of stay in NICU ≥ 5 d among the five groups also showed statistically significant differences [overall:5.4％ (5/93),1.8％ (5/281),0.6％ (1/168),0.0％ (0/32) and 2/5,x2=16.812;NICU:3.2％ (3/93),1.1％ (3/281),0.0％ (0/168),0.0％ (0/32) and 1/5; x2=1 1.294; cardiopulmonary resuscitation or ventilator therapy:2.2％ (2/93),0.7％ (2/281),0.0％ (0/168),0.0％(0/32) and 1/5,x2=10.584; asphyxiation:1.1％ (1/93),0.7％ (2/281),0.0％ (0/168),0.0％ (0/32) and 1/5,x2=9.637; NICU ≥ 5 d:3.2％ (3/93),1.1％ (3/281),0.0％ (0/168),0.0％ (0/32) and 1/5,x2=1 1.294; P＜0.05].The risks of neonatal adverse outcomes in delivery at 37-38+6 weeks were:OR=1.1(95％CI:1.0-2.1) at 37 37+6 weeks,OR=1.3 (95％CI:0.9-1.9) at 38-38+6 weeks,compared with delivery at 39-39+6 weeks.Conclnsions The percentage of repeat cesarean delivery prior to 39 weeks of gestation is high in our hospital,early termination of pregnancy would not reduce the maternal perioperative adverse outcome,but may increase the risk of neonatal adverse events.Taking into account the maternal benefit,we suggest 39 39+6 weeks of gestation as the best time of elective repeat cesarean in order to reduce the risk of neonatal adverse events.

Objective To evaluate the expression of Krüppel-like factor 8 (KLF-8) and matrix metalloproteinase 9 (MMP-9) in placentas and their relationship with the pathogenesis of preeclampsia (PE).Methods Twenty-two women with PE (mild PE:4 cases; severe PE:18 cases) who received cesarean sections in the First Affiliated Hospital of Chongqing Medical University from September 2011 to March 2012 were recruited as the PE group (n =22).And twenty women who received elective term cesarean section without perinatal complications were chosen as the control group (n =20).Placentas were collected and immunohistochemical SP method were employed to detect the localization of KLF-8 protein.KLF-8 mRNA level was determined by quantitative real-time PCR technique and western blot analysis was used to quantify KLF-8 and MMP-9 protein levels.Results (1) There was no difference of KLF-8 protein distribution in placentas of the PE group and the control group.It was mainly located in the nuclear and cytoplasm of syncytiotrophoblasts.KLF-8 immunostaining was apparently decreased in the placentas of preeclamptic women when compared with the control group.(2)The KLF-8 mRNA levels were significantly decreased in placentas of the PE group (0.69 ±0.08) compared to those of the control group (1.14 ±0.09,P ＜0.01).(3) KLF-8 and MMP-9 protein levels significantly decreased in the PE placentas (0.68 ±0.05 and 0.21 ± 0.03) when compared to the control group (0.94 ± 0.06 and 0.34 ± 0.03,respectively,P ＜ 0.01).(4) There was a positive correlation between the expression of KLF-8 and MMP-9 protein in the placentas from PE and normal pregnancies (r =0.64,P ＜ 0.01).Conclusions KLF-8 mRNA and protein levels were decreased in placentas of PE patients compared to those of normotensive women.KLF-8 protein was primarily located in the invasion-related trophoblast cells and its expression had a positive correlation with MMP-9 levels.KLF-8 might have an important role in the pathogenesis of PE by regulation of trophoblast invasion.