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.

This study was aimed to establish HPLC fingerprint of Chaenomeles speciosa for the place of production and quality control. HPLC was used to comparatively analyze 7 groups of C. speciosa crude drugs from Guizhou Zheng'an and 8 groups of C. speciosa crude drugs from other different production places. The results showed that after the mutual mode of HPLC fingerprint was set up, and the 11 common peaks were pinpointed, comparison of similar degree to crude drugs of other producing areas had been carried on. The main kinds of ingredient in C. speciosa from different places are nearly the same, but the content of each component was different. It was concluded that the method can be used as the identification and quality control of Guizhou Zheng'an C. speciosa, as well as the evaluation of herb quality and standardized planting of GAP, establishment of standard operating procedures of planting and the quality control of C. speciosa.

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.

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.

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 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 investigate the role of Shenqihuang steady plaque caps (SQH) and simvastatin on stabilization of atherosclerosis plaque in rabbits whit atherosclerosis (AS). Methods Forty-five rabbits were randomly divided into five groups: normal food group (A), instability plaque group (B), SQH group (C), simvastatin group (D), SQH add simvastatin group. A group was fed with common feeds, and B, C, D, E groups were given high cholesterol feeds. An atherosclerotic rabbit model was established by feeding high cholesterol diet supplemented for 12 weeks in B, C, D, E groups. Then C, D, E groups received corresponding drugs. At the end of 24~(th) week, the drug were injected into the aortic segments rich in plaque. Two weeks later, the concentration in serum of lipids, vascular cellular adhesion molecule-1 (VCAM-1), intercellular cellular adhesion molecule-1 (ICAM-1) and high sensitivity C reaction protein (hs-CRP), and area of plaque, the thickness of endomembrane, ttmica media, fibrous cap were detected. Results Compared with B group, the concentration of VCAM-1, ICAM-1, hs-CRP and area of plaque, the thickness of endomembrane, tunica media in C, D and E groups were lower significantly, the thickness of fibrous cap was thicker significantly (P

Objective To observe the effect of subarachnoid block with 0.5% bupivacaine at different temperatures during cesarean section.Methods 100 cases of elective cesarean section were randomly divided into room temperature group and heating group,50 cases in each group.Room temperature group: bupivacaine hydrochloride injection and glucose injection equilibrated group in a constant temperature thermostatic bath of 24 degrees thermostatic bath heating for above 30 min.Heating group: bupivacaine hydrochloride injection and glucose injection heated in the constant temperature thermostatic bath of 37 degrees thermostatic bath heatingfor above 30 min.Anesthesia was injected into the subarachnoid space at different temperatures to observe the anesthetic effect.Results The anesthesia increased rapidly, and the analgesia and muscle relaxation effects were better in the heating group than room temperature group, but the heating group had hypotension rate was higher than the room temperature group (36.0% vs.16.0%).There was no obvious difference between the incidence of adverse reactions such as nausea and vomiting in both groups.Conclusion Different temperatures of bupivacaine can be used safely for section anesthesia.The anesthesia effect of the heateding bupivacaine is faster, the anesthesia level is higher, the anesthesic and muscle relaxant effect is better.Bupivacaine at room temperature has relatively small effect on hemodynamics.

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.

To observe the efficacy and safety of mini-percutaneous nephrolithotomy with Neodymium: Yttrium Aluminum Garnet (Nd-YAG) laser in the treatment for upper urinary tract stones, from December 2005 to September 2006, 31 patients with renal stones, 15 patients with ureteral stones and 7 patients with renal and ureteral stones underwent mini-percutaneous nephrolithotomy with Nd-YAG laser by combination of rigid ureteroscope and flexible ureteroscope under B-ultrasound guidance. Clinical data including operation time, lithotripsy time, complications and stone-free rate were analyzed retrospectively. Our study showed that the percutaneous renal access (F14-F18) was successfully established under B-ultrasound guidance in all cases. Immediate phase: lithotripsy was performed in 47 cases through single tract, and in one case through two tracts. Delayed phase II lithotripsy was done in 5 cases of renal stones. Operation time ranged from 55 to 180 min with an average time of 100+/-15 min. Lithotripsy time was from 25 to 135 min and the average lithotripsy time was 65+/-11 min. No severe complications occurred in our series. Complex renal stones were cleared in 34 of 38 cases (89.5%). All ureteral stones were completely removed in 15 cases (100%). It was concluded that mini-percutaneous nephrolithotomy with Nd:YAG laser for the treatment of upper urinary tract stones by combination of rigid ureteroscope and flexible ureteroscope has the advantages of effectiveness, mini-invasion, shorter operative time and safety.