[Colonization of group B streptococcus in the semen of chronic prostatitis patients of childbearing age and its influence on perinatal outcomes].

OBJECTIVE: To investigate the colonization of group B streptococcus (GBS) in the semen of chronic prostatitis patients of childbearing age and its influence on perinatal outcomes. METHODS: This retrospective study included 592 cases of chronic prostatitis and another 472 non-prostatitis healthy males as controls. We collected semen samples from the subjects for bacterial and Ureaplasma urealyticum (UU) culture and quantitative fluorescence PCR analysis of chlamydia trachomatis (CT) and GBS. According to the results of culture, we divided the patients into a GBS-positive and a GBS-negative group and compared the perinatal outcomes among different groups of subjects. RESULTS: The rate of GBS colonization in the semen of the chronic prostatitis patients was 11.8% (70/592). Bacteria were detected in the semen of 54.4% of the patients (322/592), mainly including GBS (21.7% ï¼»70/322ï¼½) and E coli (19.9% ï¼»64/322ï¼½), and in 7.8% of the healthy controls (37/472), Staphylococcus aureus comprising 83.8% (31/37), with statistically significant difference in the rate of bacteria detection between the two groups (P < 0.01). The incidence rate of adverse perinatal outcomes in the cases of successful pregnancy was significantly higher in the GBS-positive (32.8% ï¼»19/58ï¼½) than in the GBS-negative (22.0% ï¼»29/132ï¼½) and the healthy control group (2.2% ï¼»6/271ï¼½) (P < 0.05). CONCLUSIONS: The rate of GBS colonization is significantly increased in the semen of chronic prostatitis patients of childbearing age, and so is the incidence of adverse perinatal outcomes in the spouses of GBS-positive males. Importance should be attached to normalized screening of GBS in chronic prostatitis patients and to standardized prevention and intervention as well.

Effect of lipoprotein-associated phospholipase A2 inhibitor on insulin resistance in streptozotocin-induced diabetic pregnant rats.

This paper aims to investigate the influence of lipoprotein-associated phospholipase A2 (Lp-PLA2) inhibitor, darapladib, on insulin resistance (IR) in streptozotocin (STZ)-induced diabetic pregnant rats. The rat models were divided into Control (normal pregnancy), STZ + saline (STZ-induced diabetic pregnant rats), STZ + Low-dose and STZ + High-dose darapladib (STZ-induced diabetic pregnant rats treated with low-/high-dose darapladib) groups. Pathological changes were observed by Hematoxylin-eosin (HE) and Immunohistochemistry staining. Lp-PLA2 levels were determined by enzyme-linked immunosorbent assay (ELISA). An automatic biochemical analyzer was used to measure the serum levels of biochemical indicators, and homeostatic model assessment for insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated. Western blot was applied to determine levels of inflammatory cytokines. Compared with Control group, rats in the STZ + saline group were significantly decreased in body weight, the number of embryo implantation, the number of insulin positive cells and pancreatic islet size as well as the islet endocrine cells, and high-density lipoprotein (HDL-C) level, but substantially increased in Lp-PLA2, low-density lipoprotein (LDL-C), fatty acids (FFA), serum total cholesterol (TC), triglyceride (TG) levels. Moreover, the increased fasting plasma glucose (FPG) and HOMA-IR and inflammatory cytokines but decreased fasting insulin (FINS) and ISI were also found in diabetic pregnant rats. On the contrary, rats in the darapladib-treated groups were just opposite to the STZ + saline group, and STZ + High-dose group improved better than STZ + Low-dose group. Thus, darapladib can improve lipid metabolism, and enhance insulin sensitivity of diabetic pregnant rats by regulating inflammatory cytokines.

[Long noncoding RNA HOTAIR modulates the function of trophoblast cells in pre-eclampsia].

OBJECTIVE: To investigate the expression of long noncoding RNA (lncRNA) HOTAIR in pre-eclampsia (PE) placentas and its effect on trophoblast cells proliferation, invasion, and apoptosis. METHODS: The expression of HOTAIR was determined by qPCR for 23 severe PE and 23 normal pregnant women. qRT-PCR was used to detect the efficiency of overexpression and knock-down after the HTR-8/SVneo cells were transfected with HOTAIR overexpression and siRNAs targeting HOTAIR for 24-48 h, respectively. MTT and colony formation assays were used to test the proliferation of HTR-8/SVneo cells transfected. Transwell assay was used to show the migration and invasion ability of HTR-8/SVneo cells transfected. Flow cytometry assay was used to detect the cell apoptosis rate after treatment. Western blot assay was applied to detect the expression level of apoptotic proteins Caspase-3 and BCL-2. RESULTS: The level of HOTAIR in severe pre-eclampsia groups was significantly increased compared to normal pregnant placentas (P<0.05). The expression of HOTAIR in HTR-8/SVneo cells after transfected with pcDNA-HOTAIR was 51. 27-fold than that of control; while inhibition of HOTAIR was more than 95% in si-HOTAIR than that of control. According to the MTT and colony formation assays, we found that cells proliferation rate of cells were significantly decreased in overexpression HOTAIR group while increased in si- HOTARI group when compared with control, respectively. The transwell assay showed that the invasive capacity of HTR-8/SVneo cells in cells transfected with pcDNA-HOTAIR decreased, while increased in si-HOTAIR transfected cells when compared with that of control. The apoptosis rate in cells transfected with HOTAIR overexpression was apparently more than that of control, while less in cells treated with si-HOTAIR. Western blot assay showed that the Caspase-3 showed an obvious increase in HOTAIR overexpression group while decreased in si-HOTAIR group. And BCL-2 presented an opposite trend. CONCLUSION: HOTAIR is probably involved in the onset of preeclampsia by regulating proliferation, invasion and apoptosis of trophoblast cells.

Peptidome analysis of human skim milk in term and preterm milk.

The abundant proteins in human milk have been well characterized and are known to provide nutritional, protective, and developmental advantages to both term and preterm infants. Due to the difficulties associated with detection technology of the peptides, the expression of the peptides present in human milk is not known widely. In recent years, peptidome analysis has received increasing attention. In this report, the analysis of endogenous peptides in human milk was done by mass spectrometry. A method was also developed by our researchers, which can be used in the extraction of peptide from human milk. Analysis of the extracts by LC-MS/MS resulted in the detection of 1000-3000Da peptide-like features. Out of these, 419 peptides were identified by MS/MS. The identified peptides were found to originate from 34 proteins, of which several have been reported. Analysis of the peptides' cleavage sites showed that the peptides are cleaved with regulations. This may reflect the protease activity and distribution in human body, and also represent the biological state of the tissue and provide a fresh source for biomarker discovery. Isotope dimethyl labeling analysis was also used to test the effects of premature delivery on milk protein composition in this study. Differences in peptides expression between breast milk in term milk (38-41weeks gestation) and preterm milk (28-32weeks gestation) were investigated in this study. 41 Peptides in these two groups were found expressed differently. 23 Peptides were present at higher levels in preterm milk, and 18 were present at higher levels in term milk.

[Effect of transforming growth factor ß1 on the expression of matrix metalloproteinase 9, tissue inhibitor of metalloproteinase 1 and nuclear factor kappa B signalling pathway in the human amniotic cells WISH].

OBJECTIVE: To investigate the effect of transforming growth factor ß1 (TGF-ß1) on the expression of matrix metalloproteinase 9 (MMP-9), tissue inhibitor of metalloproteinase 1 (TIMP-1), nuclear factor kappa B (NF-κB) and the possible signalling pathways in human amniotic cells WISH. METHODS: The WISH cell line was cultured. WISH cells were added with TGF-ß1 of different concentrations (0, 2, 10 and 20 ng/ml, respectively) for 24 hours. Then, reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot. RESULTS: (1) The profile of TIMP-1 mRNA (0.413 ± 0.036, 0.623 ± 0.058, 1.392 ± 0.124, 1.387 ± 0.102) in WISH cells elevated when the concentration of TGF-ß1 increased (0, 2, 10, 20 ng/ml). In accordance with TIMP-1 mRNA, the expression of TIMP-1 also elevated with the increase of TGF-ß1 (0.357 ± 0.031, 0.596 ± 0.048, 1.243 ± 0.097 and 1.359 ± 0.121, respectively). And when 2, 10 or 20 ng/ml of TGF-ß1 was added, the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-ß1 was added (P < 0.05). (2) In contrast with TIMP-1, MMP-9 mRNA (1.325 ± 0.056, 0.987 ± 0.081, 0.610 ± 0.034, 0.347 ± 0.023) in WISH cells decreased when the concentration of TGF-ß1 increased (0, 2, 10, 20 ng/ml). The MMP-9 protein (1.119 ± 0.064, 1.008 ± 0.052, 0.578 ± 0.041, 0.401 ± 0.015) also decreased with the increase of TGF-ß1. And when 2, 10 or 20 ng/ml of TGF-ß1 was added, the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-ß1 was added (P < 0.05). (3) The NF-κB protein (1.423 ± 0.065, 1.116 ± 0.045, 0.796 ± 0.041, 0.359 ± 0.021) was significantly reduced with the increase of TGF-ß1 (0, 2, 10, 20 ng/ml;P < 0.05). CONCLUSIONS: The mRNA and protein expression of TIMP-1 decreased when TGF-ß1 was low in WISH cells, whereas those of MMP-9 elevated when TGF-ß1 was low. The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane. And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.

A novel bridge between oxidative stress and immunity: the interaction between hydrogen peroxide and human leukocyte antigen G in placental trophoblasts during preeclampsia.

OBJECTIVE: The aim of this study was to investigate a possible association between hydrogen peroxide and human leukocyte antigen G (HLA-G) in preeclampsia. STUDY DESIGN: We explored the correlation between HLA-G and hydrogen peroxide in preeclamptic (n = 30) and normotensive (n = 30) placentas, which was confirmed by in vitro experiments. Furthermore, RNA interference was used to explore the influence of HLA-G on the proliferation, apoptosis and invasion of HTR-8/SVneo cells when exposed to hydrogen peroxide. RESULTS: We found an inverse correlation between hydrogen peroxide and HLA-G expression in preeclamptic placentas. High levels of hydrogen peroxide could down-regulate HLA-G expression in HTR-8/SVneo. Compared with HLA-G knockdown HTR-8/SVneo, increased proliferation inhibition, higher apoptosis, and decreased cell invasion were found in the cell expressing HLA-G when exposed to hydrogen peroxide. CONCLUSION: Our findings highlight that high levels of hydrogen peroxide can down-regulate HLA-G expression in trophoblasts during preeclampsia and trophoblasts expressing HLA-G are vulnerable to oxidative stress.

[Effect of epidermal growth factor on the expression of matrix metalloproteinase-9 and the signalling pathways involved in the trophoblast cell line JEG-3].

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 10 ng/ml EGF), EGF+ inhibitors group (exposure to 10 ng/ml EGF+ 20 ng/ml SB203580 or exposure to 10 ng/ml EGF+ 10 ng/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-κB), 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-9 mRNA were increased by various concentrations of EGF (0, 1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0.567±0.056), 10 ng/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 p38MAPK 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-κB (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-κB (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.

[Evaluation the efficacy and safety of estradiol and drospirenone tablets in the treatment of menopausal symptoms among postmenopausal Chinese healthy women:a randomized, multi-center, double-blind, placebo-controlled clinical study].

OBJECTIVE: To study the efficacy and safety of estradiol and drospirenone tablets (Angeliq) in treatment of menopausal symptoms among postmenopausal Chinese healthy women. METHODS: Total 244 postmenopausal Chinese healthy women who had moderate to severe hot flushes were randomly assigned into estradiol and drospirenone (observation group, n = 183) or placebo group (n = 61) by the ratio of 3:1 for 16 weeks in this randomized multi-center double-blind placebo-controlled study. During the trial, the follow-up visits were conducted at week 4, 8, 12, 16 of treatment and 2 weeks after treatment respectively. Height, weight, vital signs, hot flushes, other relevant menopausal symptoms and vaginal bleeding were observed in each follow-up visit, while the clinical global impression scale was assessed at 16 weeks as well. RESULTS: It showed that hot flushes were reduced significantly more in observation group than that in placebo group (P < 0.01), although both treatments were effective. The absolute values of mean severity index of total hot flushes decreased by -0.6 ± 0.5 in observation group and -0.4 ± 0.4 in placebo group from baseline respectively, which reached significant difference (P < 0.05). However, the absolute values of mean severity index of moderate to severe hot flushes decreased by -0.6 ± 0.8 in observation group and -0.3 ± 0.6 in placebo group from baseline respectively, which had no significant difference (P > 0.05). After 16 weeks treatment, it also showed that estradiol and drospirenone had significant better efficacy than placebo on moderate to severe sweating, vaginal dryness and clinical global impression scale (P < 0.01). During the trial, blood pressure in observation group was stable. The rate of vaginal bleeding in observation group was higher than that in the placebo group, especially during the week 4 to week 8 when 48.9% (87/178) in observation group and 10.7% (6/56) in placebo group of patients bled. Although the cumulative amenorrhea rate of observation group was lower than that of placebo group in each cycle (28 days), it increased gradually along with duration of the treatment. The commonest adverse event in observation group was breast tenderness which accounted for 12.0% (22/183). The level of serum potassium was in the normal range in observation group mostly.Meanwhile, the other adverse events rate was low. Serious adverse events reported in this trial were assessed as not study drug related or as unlikely study drug related. CONCLUSION: Estradiol and drospirenone tablets which could effectively alleviate menopausal symptoms in postmenopausal Chinese healthy women is a novel hormone replacement therapy regimen with high safety and efficacy.

[Effect of hydrogen peroxide on human leukocyte antigen-G expression in placental trophoblasts in pre-eclampsia].

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 micromol/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) nmolxmg(-1)xprot(-1) vs (62+/-18) nmol.mg(-1)xprot(-1), P<0.05]. (2) The expression of HLA-G protein in placenta of the pre-eclampsia group was reduced by 88% 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.

[Effect of endoplasmic reticulum stress-responsive protein glucose-regulated protein 78, 94 and endoplasmic reticulum apoptosis factor caspase-12 in trophocyte on the pathogenesis of preeclampsia].

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.

[A multi-center study to evaluate the dynamic changes of uterine artery and umbilical artery flow in a normal pregnancy and hypertensive disorders in pregnancy].

OBJECTIVE: To investigate the dynamic changes of uterine artery and umbilical artery in the first, second, and third trimester of normal pregnancy and hypertensive disorders in pregnancy (HDP). METHODS: A multi-center prospective study was conducted on 1098 women with normal singleton pregnancies at the first prenatal visit in the Second West China Hospital of Sichuan University, First Affiliated Hospital with Nanjing Medical University, Beijing Obstetrics and Gynecology Hospital Affiliated to Capital Medical University, Wuhan Union Hospital Affiliated to Medical School of Huazhong University of Science and Technology and Renji Hospital Affiliated to Medical School of Shanghai Jiao Tong University from April 2005 to July 2006, with the average age of (28.3 ± 3.3). The pulsatility indices (PI), resistance indices (RI) and systolic to diastolic velocity ratios (S/D) of uterine artery and umbilical artery were measured for all subjects in the first (10th-14th gestational weeks), second (20th-26th gestational weeks) and third trimester (30th-36th gestational weeks), respectively. In this longitudinal study, women who developed HDP were classified into HDP group, and the rest into normal pregnancy group. RESULTS: (1) Among the 1098 pregnant women, 44 developed HDP during the index pregnancy, including 20 gestational hypertension, 15 mild pre-eclampsia and 9 severe pre-eclampsia, giving an incidence of 4.17%, and the rest 1054 were normal until delivery. (2) In the normal pregnancy group, the RI, PI and S/D of uterine artery were decreased with the progress of pregnancy (RI: 0.64, 0.57, 0.50; PI: 1.24, 0.98, 0.80; S/D: 3.26, 2.58, 2.20; P < 0.01). However, the above indices showed an increasing trend with the increase of gestations in the HDP group (RI: 0.55, 0.67, 0.64; PI: 1.22, 1.36, 1.20; S/D: 3.18, 3.41, 3.05; P < 0.01). In the second and third trimester, the RI, PI and S/D of uterine artery in the HDP group were higher than those in the normal pregnancy group (P < 0.01). (3) In the normal pregnancy group, the RI, PI and S/D of the umbilical artery decreased from the second to the third trimester (RI: 0.71 and 0.58; PI: 1.16 and 0.87; S/D: 3.58 and 2.48; P < 0.01). However, no significant difference was found in the RI, PI and S/D value of umbilical artery in the second and third trimester between the normal and HDP group (RI: 0.71 and 0.63; PI: 1.20 and 0.95; S/D: 3.71 and 2.69; P > 0.05, respectively), despite the decreasing trend in the HDP group. CONCLUSIONS: In uncomplicated pregnancies, the blood flow resistance of uterine artery decreases and the end-diastolic blood flow of uterine artery increases with the progress of pregnancy. However, in pregnant women with HDP, the blood flow resistance of uterine artery increases significantly with the increase of gestations which shows significant difference to that in normal pregnancies. The blood flow resistance of umbilical artery decreases in both normal and HDP pregnant women with the increasing gestational age.

[Vaginal microflora and relevant factors in puerperium].

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 postpartum women from 7 hospitals in China were enrolled. Questionnaire survey, gynecological examination and Nugent score of vaginal smear and microbial spectrum study of the vaginal flora were completed. RESULTS: (1) According to the Nugent 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, 25 (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 = 0.185), 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.

[Multi-center study of motherwort injection to prevent postpartum hemorrhage after caesarian section].

OBJECTIVE: To evaluate the efficacy and safety of motherwort (herba leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. METHODS: The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apr 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 mg uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort+oxytocin: 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U+5% glucose 500 ml intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell (RBC) from prepartum to postpartum. (4) Adverse reaction. RESULTS: (1) The mean amount of blood loss during operation were (368+/-258) ml in group of motherwort, (255+/-114) ml in group of motherwort+oxytocin and (269+/-141) ml in group of oxytocin, which exhibited significant difference among three groups (P<0.01). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2, 6, 12, 24, 48 hours after CS. (2) The amount of blood loss of postpartum at 24 hours were (480+/-276) ml in group of motherwort, (361+/-179) ml in group of motherwort+oxytocin, (381+/-179) ml in group of oxytocin, which showed significant difference among 3 groups (P<0.01). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort+oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort+oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC (0.3+/-0.5)x10(12)/L and Hb (9+/-13) g/L in group of motherwort, RBC (0.2+/-0.4)x10(12)/L and Hb (6+/-10) g/L in group of motherwort+oxytocin and RBC (0.2+/-0.4)x10(12)/L and Hb (7+/-30) g/L in group of oxytocin respectively. However, the comparison of different value of RBC and Hb in group of oxytocin and motherwort+oxytocin showed significant difference (P<0.05). (5) Two cases with allery reaction was observed. CONCLUSION: It is safe and efficacious that combined use of motherwort injection and oxytocin was to prevent postpartum hemorrhage during or after caesarian section.

Correlation of serum vitamin D, adipose tissue vitamin D receptor, and peroxisome proliferator-activated receptor γ in women with gestational diabetes mellitus.

BACKGROUND: Gestational diabetes mellitus (GDM) is a common complication during pregnancy. Obesity and overweight are closely related to metabolic diseases and diabetes. However, the role of adipose tissue in the pathogenesis of GDM remains to be studied. The aim of this study was to investigate the correlation of vitamin D (VD) levels, VD receptor (VDR), and peroxisome proliferator-activated receptor γ (PPARγ) expression with GDM in overweight or obese women. METHODS: One hundred and forty pregnant women with full-term single-birth cesarean-section were selected as the study subjects and grouped (70 GDM women, including 35 non-overweight/non-obese women [group G1] and 35 women with overweight or obesity [group G2]; 70 pregnant women with normal glucose tolerance, including 35 non-overweight/non-obese women [group N1] and 35 overweight/obese women [group N2]). The levels of serum VD, blood biochemistry, and adiponectin were compared in these women. Subcutaneous adipose tissue was isolated from the abdominal wall incision. VDR and PPARγ messenger RNA (mRNA) transcript levels in these adipose tissues were quantified by real-time polymerase chain reaction. The differences between the levels of PPARγ protein and phosphorylated PPARγ Ser273 were detected by Western blotting. RESULTS: The serum VD level of GDM women was lower in comparison to that of women with normal glucose tolerance (G1 vs. N1: 20.62 ±â€Š7.87 ng/mL vs. 25.85 ±â€Š7.29 ng/mL, G2 vs. N2: 17.06 ±â€Š6.74 ng/mL vs. 21.62 ±â€Š7.18 ng/mL, P < 0.05), and the lowest in overweight/obese GDM women. VDR and PPARγ mRNA expression was higher in the adipose tissues of GDM women in comparison to that of women with normal glucose tolerance (VDR mRNA: G1 vs. N1: 210.00 [90.58-311.46] vs. 89.34 [63.74-159.92], G2 vs. N2: 298.67 [170.84-451.25] vs. 198.28 [119.46-261.23], PPARγ mRNA: G1 vs. N1: 100.72 [88.61-123.87] vs. 87.52 [66.37-100.04], G2 vs. N2: 117.33 [100.08-149.00] vs. 89.90 [76.95-109.09], P < 0.05), and their expression was the highest in GDM + overweight/obese women. VDR mRNA levels positively correlated with the pre-pregnancy body mass index (BMI), pre-delivery BMI, fasting blood glucose (FBG), homeostasis model assessment of insulin resistance (HOMA-IR), and PPARγ mRNA while it negatively correlated with the VD and the adiponectin levels (r = 0.395, 0.336, 0.240, 0.190, 0.235, -0.350, -0.294, respectively, P < 0.05). The degree of PPARγ Ser273 phosphorylation increased in obese and GDM pregnant women. PPARγ mRNA levels positively correlated with pre-pregnancy BMI, pre-delivery BMI, FBG, HOMA-IR, serum total cholesterol, triglyceride, free fatty acid, and VDR mRNA, while it negatively correlated with the VD and adiponectin levels (r = 0.276, 0.199, 0.210, 0.230, 0.182, 0.214, 0.270, 0.235, -0.232, -0.199, respectively, P < 0.05). CONCLUSIONS: Both GDM and overweight/obese women had decreased serum VD levels and up-regulated VDR and PPARγ mRNA expression in adipose tissue, which was further higher in the overweight or obese women with GDM. VD may regulate the formation and differentiation of adipocytes through the VDR and PPARγ pathways and participate in the occurrence of GDM.

Proteomic analysis of proteins differentially expressed in preeclamptic trophoblasts.

AIMS: To identify differential trophoblastic proteins associated with preeclampsia (PE) by proteomic analysis. METHODS: We isolated and purified placental trophoblasts from normotensive pregnant women and patients with PE by a continuous Percoll gradient. The expression of proteins was determined by sliver staining after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Proteins of interest were identified using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). RESULTS: The overall trophoblastic protein expression patterns in preeclamptic and corresponding normotensive placentas were quite similar except for some areas. Of 34 differentially expressed protein spots (p < 0.05 by paired t-test), seven differential proteins from nine spots were identified by MALDI-TOF-MS. The expression of the following proteins was repressed (p < 0.01): disulfide isomerase ER-60, peroxiredoxin 2, and Delta3,5-Delta2,4-dienoyl-CoA isomerase. Four proteins (protein disulfide isomerase precursor, endoplasmic reticulum resident protein, dihydrolipoyl dehydrogenase and TIM21-like protein) were found to be significantly upregulated in PE (p < 0.01). CONCLUSION: We identified several proteins with significant altered expression in PE using 2D-PAGE. This method is a powerful technique with which to search for not only quantitative but also qualitative changes in a biological process of interest.

Genetic variation of rs3811463 is associated with gestational diabetes mellitus susceptibility.

Gestational diabetes mellitus (GDM) is a growing health concern, and it increases the risk of adverse pregnancy outcomes with substantial long-term adverse health impacts on mothers and their offspring. Several studies have revealed specific associations between genetic variants and the risk of GDM. Single nucleotide polymorphisms (SNPs) are the major type of genetic variation in humans. Let-7 microRNA targets are enriched for genes containing SNPs associated with glucose metabolism, including Lin28. In the present study, the effect of T/C variants of rs3811463 (a SNP located near to the let-7 binding site in Lin28) on GDM risk was investigated. A GDM rat model was successfully constructed using a high fat diet and streptozotocin injection, and the primary skeletal muscle cells were isolated. The cell transfection results demonstrated that rs3811463-T/C significantly affected the glucose uptake and insulin sensitivity. Reverse transcription-quantitative polymerase chain reaction analysis indicated that the C allele at rs3811463 regulated the expression of glucose metabolism-associated genes insulin-like growth factor two binding protein 2 and glucokinase. Western blot analysis data revealed that replacement of the T allele by the C allele at rs3811463 modulated the protein level of Sirtuin 1. Taken together, it was concluded that the let-7/Lin28 axis regulated glucose uptake and insulin sensitivity by modulating the expression of glucose metabolism-associated proteins. These findings provide novel evidence on the association between genetic variations of rs3811463 and GDM susceptibility.

Intravaginal Misoprostol for Cervical Ripening and Labor Induction in Nulliparous Women: A Double-blinded, Prospective Randomized Controlled Study.

BACKGROUND: In China, no multicenter double-blinded prospective randomized controlled study on labor induction has been conducted till now. This study is to evaluate the efficacy and safety of intravaginal accurate 25-µg misoprostol tablets for cervical ripening and labor induction in term pregnancy in nulliparous women. METHODS: This was a double-blinded, prospective randomized controlled study including nulliparous women from 6 university hospitals across China. Subjects were randomized into misoprostol or placebo group with the sample size ratio set to 7:2. Intravaginal 25-µg misoprostol or placebo was applied at an interval of 4 h (repeated up to 3 times) for labor induction. Primary outcome measures were the incidence of cumulative Bishop score increases ≥3 within 12 h or vaginal delivery within 24 h. Safety assessments included the incidences of maternal morbidity and adverse fetal/neonatal outcomes. RESULTS: A total of 173 women for misoprostol group and 49 women for placebo were analyzed. The incidence of cumulative Bishop score increases ≥3 within 12 h or vaginal delivery within 24 h was higher in the misoprostol group than in the placebo (64.2% vs. 22.5%, relative risk [RR]: 2.9, 95% confidence interval [CI]: 1.4-6.0). The incidence of onset of labor within 24 h was significantly higher in the misoprostol group than in the placebo group (48.0% vs. 18.4%, RR: 2.6, 95% CI: 1.2-5.7); and the induction-onset of labor interval was significantly shorter in the misoprostol group (P = 0.0003). However, there were no significant differences in the median process time of vaginal labor (6.4 vs. 6.8 h; P = 0.695), incidence (39.3% vs. 49.0%, RR: 0.8, 95% CI: 0.4-1.5) and indications (P = 0.683) of cesarean section deliveries, and frequencies of maternal, fetal/neonatal adverse events between the groups. CONCLUSION: Intravaginal misoprostol 25 µg every 4 h is efficacious and safe in labor induction and cervical ripening.

[Apoptosis gene expression profiling of placental trophoblast cells in patients with pregnancy induced hypertension].

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 micro m(2) was 1.584 in the PIH group and 0.032 in the control group with significant difference between the two groups (P < 0.01). (2) Ten differently expressed apoptosis genes were obtained through gene chip test (occupy 5% of total apoptosis gene in the gene chip). There was a significant decrease of apoptosis gene expression in all of PIH patient placental tissues (i.e a ratio of cy5/cy3 less than 1). Among them, there were genes that possess significant anti-apoptosis functions (including SFRP(2), IAP(2), DHCY24 and ATPIA1). CONCLUSIONS: Remarkable apoptosis was found in placental trophoblast cells of PIH patients. Significant decrease in genes with anti-apoptosis functions can result in the apoptosis of placental trophoblast cells and thus contributes to the pathogenesis of PIH.

Fasting plasma glucose at 24-28 weeks to screen for gestational diabetes mellitus: new evidence from China.

OBJECTIVE: To evaluate the usefulness of a fasting plasma glucose (FPG) at 24-28 weeks' gestation to screen for gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: The medical records and results of a 75-g 2-h oral glucose tolerance test (OGTT) of 24,854 pregnant women without known pre-GDM attending prenatal clinics in 15 hospitals in China were examined. RESULTS: FPG cutoff value of 5.1 mmol/L identified 3,149 (12.1%) pregnant women with GDM. FPG cutoff value of 4.4 mmol/L ruled out GDM in 15,369 (38.2%) women. With use of this cutoff point, 12.2% of patients with mild GDM will be missed. The positive predictive value is 0.322, and the negative predictive value is 0.928. CONCLUSIONS: FPG at 24-28 weeks' gestation could be used as a screening test to identify GDM patients in low-resource regions. Women with an FPG between ≥4.4 and ≤5.0 mmol/L would require a 75-g OGTT to diagnose GDM. This would help to avoid approximately one-half (50.3%) of the formal 75-g OGTTs in China.

[The relationship among a portable gas chromatography, organoleptic method and portable sulfide detector to diagnose halitosis].

PURPOSE: To evaluate the relationship between a portable gas chromatography(GC-SCS) and other methods to diagnose halitosis. METHODS: Fifty-five systemic healthy subjects were involved in the study.Halitosis intensity was examined by two experienced judges using organoleptic method. Halimeter was used to evaluate the concentrations of volatile sulfur compounds while GC-SCS was used to tell the volume of each volatile sulfur compound. SPSS11.5 software package was used for Pearson and Spearman correlation analysis. RESULTS: The results showed that the concentration of hydrogen sulfide, methyl mercaptan and total VSCs measured by GC-SCS was significantly related to the organoleptic scores with the correlational coefficients of 0.486, 0.529 and 0.491, respectively (P<0.01). There was no statistically significant relationship between the dimethyl sulfide values and the organoleptic scores. The VSCs value measured by Halimeter was significantly related to the concentration of hydrogen sulfide, methyl mercaptan and total VSCs measured by GC-SCS with the correlational coefficients of 0.458, 0.522 and 0.436, respectively (P<0.01). CONCLUSIONS: GC-SCS relates well with organoleptic method and Halimter. It is suggested that GC-SCS may be a good way to diagnose halitosis.