In order to detect the expression of RhoA in placenta from normal pregnancy and preeclampsia and evaluate the role of RhoA in preeclampsia, the expression of RhoA in placenta collected from 40 preeclampsia patients and 20 normotensive controls was determined by immunohistochemistry and RT-PCR. RhoA was found in syncytiotrophoblasts and cytotrophoblasts. The mean density of RhoA protein in placental tissues of mild and severe preeclampsia groups was significantly higher than that in normal pregnancy. The expression level of RhoA mRNA in mild and severe preeclampsia groups was significantly higher than that of normal pregnancy. Increased expression of RhoA in placental tissues might play an important role in the pathogenesis of preeclampsia.

A new nested-polymerase chain reaction (nested-PCR) assay was developed to detect human parvovirus B19 DNA corresponding to the nonstructural protein in clinical specimens in a routine diagnostic laboratory. The sensitivity of this highly specific assay was up to 0.005 fg of B19 DNA. Parvovirus B19 was identified in sera of 20 pregnant women with abnormal pregnant outcome. Among these 20 cases, intrauterine parvovirus infection did exist in 7 pregnant women because parvovirus B19 DNA was detected in the pregnant tissues of them such as placenta tissues, chorionic villi, amniotic fluid, fetal spleen, liver and abdominal fluids.
DNA, Viral/*analysis ; Parvoviridae Infections/*virology ; Parvovirus B19, Human/genetics ; Parvovirus B19, Human/*isolation & purification ; Placenta/virology ; Pregnancy Complications, Infectious/diagnosis ; Pregnancy Complications, Infectious/*virology ; Viral Nonstructural Proteins/*analysis

In order to observe the effect of ursodeoxycholic acid (UDCA) in the treatment of intrahepatic cholestasis of pregnancy (ICP), 68 patients with ICP were equally divided into treatment group and control group at random. The patients in treatment group were administered with UDCA 300 mg three times every day and those in control group received a combination of 10% glucose, Vitamin C and Inosine. Itching scores, serum ALT and total bile acids (TBA) were measured before, during and after treatment. The results showed that as compared with those before treatment, itching scores, serum ALT and TBA were significantly reduced after treatment (P < 0.05). The occurrences of premature labor, fetal asphyxia and meconium staining in amniotic fluid were significantly lower in treatment group than in control group (P < 0.05). It was suggested that UDCA was an effective drug in the treatment of ICP.

The effects of leptin on cytotrophoblast proliferation and invasion activity were investigated. Immunohistochemistry was used to determine the placental expression of leptin in first-trimester pregnancy. By using RT-PCR and quantitative real-time PCR, the expression of leptin in cytotrophoblast and the effect of leptin on cytotrophoblast secretion were detected. The potential of cell proliferation, invasiveness and migration was assessed by MTT, Transwell invasion assay and migration assay respectively when the cytotrophoblast was cultured with different concentrations of leptin. The results showed that: (1) Leptin was distributed diffusely around cell membrane, in cytoplasma, and on nuclear membrane of cytotrophoblast; (2) Leptin mRNA was expressed in cytotrophoblast. Ten ng/mL leptin could promote the secretion of cytotrophoblast significantly (P<0.01); (3) After culture with different concentrations of leptin for 24 h or longer, the proliferation of cytotrophoblast was inhibited, while in 24 h leptin could promote cytotrophoblast invasion and migration. Leptin at a concentration of 500 ng/mL could promote cytotrophoblast invasiveness and migration significantly as compared with controls (P<0.05). It was suggested that leptin could inhibit cytotrophoblast proliferation, and promote cytotrophoblast invasion and migration activity.

To investigate the expression of osteopontin (OPN) and its receptor integrin alphanubeta3 in the placental tissue from pregnant women complicated with preeclampsia, the expression of OPN and alphanubeta3 in the placenta of the pregnant women with preeclampsia and healthy pregnant women was detected by immunohistochemistry, Western blotting and RT-PCR. Our results showed that OPN and alphanubeta3 protein were expressed in the placenta from normal pregnant woman and those with preeclampsia. OPN was located in the placental syncytiotrophoblasts and the cytoplasm of capillary endothelial cells and integrin alphanubeta3 was mainly expressed on the surface of trophoblast cells. Expression of OPN and integrin alphanubeta3 in the placental tissue from preeclampsia subjects was significantly lower than that from the control group (P<0.05). Compared with the control group, expression of OPN in the placental tissue from preeclampsia group was significantly lower (P<0.05) but there was no significant difference in the expression of alphanu and beta3 between the preeclampsia group and the controls. It is concluded that OPN and its receptor integrin alphanubeta3 may be involved in the pathogenesis of preeclampsia.

This study examined the association of expression of vascular endothelial growth factor (VEGF), a promoter of angiogenesis, with endothelium dysfunction in preeclampsia. The level of VEGF protein and mRNA in the placenta and peripheral blood samples of 30 preeclampsia patients and 30 normotensive pregnant women was measured by immunohistochemistry, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. VEGF expression in the human umbilical vein endothelial cells (HUVECs) was blocked by small interfering RNAs (siRNAs). The monolayer barrier function of HUVECs was determined by measuring the fluorescence intensity of BSA that crossed the HUVEC monolayers. The cell proliferation and cell-secreted nitric oxide (NO) level were detected by MTT method and nitrate reductase assay, respectively. The results showed that VEGF was expressed in the syncytiotrophoblasts and endothelial cells of vessels and capillaries in the placenta tissue. The serum level of VEGF in the preeclampsia patients was significantly decreased as compared with that in normal pregnant subjects, although VEGF mRNA expression in the placenta tissue of preeclampsia patients remained still high. Moreover, VEGF deficit could lead to endothelium cell dysfunction, and the administration of VEGF could protect endothelium cells from injury. It was concluded that lack of VEGF contributes to endothelium dysfunction, which may lead to the occurrence and development of preeclampsia.

ObjectiveTo investigate the effects of vascular endothelial growth factor (VEGF) and dexamethasone on mRNA expressions of surfactant protein B (SP-B) and transforming growth factor-β1 (TGF-β1) of type Ⅱ alveolar epithelial cell (AECⅡ). MethodsAECⅡ were isolated and purified from fetal rat lung tissues,then cultured with different dose of VEGF (25,50 and 100 ng/ml) and dexamethasone (25,50,100 and 200 nmol/ml).The mRNA levels of SP-B and TGF-β1 were detected by real-time quantitative polymerase chain reaction (RT-PCR) and expression of TGF-β1 protein was detected by immunocytochemistry.ANOVAor q-test wasappliedtocompare the difference among groups.ResultsCompared with control group,SP-B mRNA levels in 25 ng/ml VEGF group and 25,50,100 and 200 nmol/ml dexamethasone groups were higher (13.500±3.172,3.547±0.690,5.219±0.782,3.493±0.335,and 3.981 ± 1.133 vs 1.001 ± 0.059,q=-5.286,-4.943,- 7.228,- 9.906 and - 3.525 respectively,P＜0.05) ; TGF-β1 mRNA expression of 25 ng/ml VEGF group,50,100 and 200 nmol/ml dexamethasone group was lower (0.451 ± 0.078,0.579±0.019,0.422 ± 0.020 and 0.769 ± 0.025 vs 1.019±0.226,q=4.110,3.356,4.551 and 1.901 respectively,P＜0.05) ; other groups had no significant differences compared with control group (P＞0.05).Immunocytochemistry showed that the positive rate of TGF-β1 expression in 25 ng/ml VEGF,50,100 and 200 nmol/ml dexamethasone group was 23％,26％,22％ and 29％,respectively,while in the control group,the expression of TGF-β1 was positive in most of the AECⅡ (80％).ConclusionsBoth VEGF and dexamethasone could increase the expression of SP-B at mRNA level at appropriate concentrations.At the same time,the expression of TGF-β1 is inhibited.It is suggested that both VEGF and dexamethasone might increase the mRNA expression of SP-B by inhibiting the expression of TGF-β1.

Objective:The treatment of late pregnancy complicated with utedne leiomyoma was investigated.Methods:193 Cases of Iate pregnancy complicated with uterine leiomyoma from January 2003 to August 2008 were recruited in our hospital.According to the delivery route,size and subtype of fibroid,blood loss,operation hours and postoperative inpatient period were compared.Results:104 cases of pregnancy complicated with uterine leiomyoma were diagnosed before cesarean section(CS).No significant differences on blood losses and operation hours were found between CS group and CS+myomectomy group(P>0.05).The operation heurs of leiomyoma in corpus uteri was significantly shorter than leiomyoma in lower uterine segment and cervix(P=0.007).Leiomyoma bigger than 8 cm needed significantly Ionger operative hours and lose more blood than the smaller leiomyoma.Operation hours,blood loss and postoperative inpatient period were significantly different between submucous leiomyoma and subserosal leiomyoma(P<0.05).Conclusions:Pregnancy complicated with uterine leiomyoma should be diagnosed as early as possible.During cesarean section on when leiomyoma is bigger than 8 cm,locating at lower uterine segment or cervix or submucous,the treatment should be cautious.

Objective To investigate the correlation between 24 h urinary protein quantitation and pregnancy outcome in patients with pre-eclampsia.Methods A total of 332 pre-eclampsia patients were selected in Tongji Hospital from January 2014 to December 2016.The patients were divided into microalbuminuria group(24 h urinary protein quantification ＜ 0.3 g,n =46),mild proteinuria group (0.3 g ≤ 24 h urinary protein quantification ＜ 2.0 g,n =98),moderate proteinuria group (2.0 g ≤ 24 h urinary protein quantification ＜ 5.0 g,n =71) and severe proteinuria group(24 h urinary protein quantification ≥ 5.0 g,n =117) according to the results of 24 h urinary protein quantification.The pregnancy outcomes were compared between the four groups.Results The 24 h urinary protein quantification and the serum creatinine,urea nitrogen,uric acid levels in the mild proteinuria group,moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the microalbuminuria group (P ＜ 0.05).The 24 h urinary protein quantification and serum urea nitrogen,uric acid levels in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in serum creatinine level between the two groups (P ＞ 0.05).The 24 h urinary protein quantification,serum creatinine,urea nitrogen and uric acid levels in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the gestational week was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The 24 h urinary protein quantification in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the gestational week and serum creatinine,urea nitrogen,uric acid levels between the two groups (P ＞0.05).There was no significant difference in the rates of cesarean section and spontaneous labor between the four groups (P ＞0.05).The rate of induced labor in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the mild albuminuria group and the microalbuminuria group (P ＜ 0.05).There was no significant difference in the rate of induced labor between the mild proteinuria group and the microalbuminuria group (P ＞ 0.05).There was no significant difference in the rate of induced labor between the severe proteinuria group and the moderate proteinuria group (P ＞ 0.05).The incidence of complications in microalbuminuria group,mild proteinuria group,moderate proteinuria group and severe proteinuria group was 30.43％ (14/46),47.96％ (47/98),74.65％ (53/71) and 74.36％ (87/117) respectively;the incidence of complications in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the microalbuminuria group and the mild albuminuria group (P ＜ 0.05),but there was no significant difference in the incidence of complications between microalbuminuria group and mild albuminuria group (P ＞ 0.05),there was no significant difference in the incidence of complications between the moderate proteinuria group and the severe proteinuria group (P ＞0.05).The incidences of premature birth and neonatal asphyxia in the mild proteinuria group were significantly higher than that in the microalbuminuria group (P ＜ 0.05),and the body mass of the neonates was significantly lower than that in the microalbuminuria group (P ＜0.05),but there was no significant difference in the perinatal mortality rate and the incidences of fetal growth restriction(FGR) and poor neonatal resuscitation between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and the perinatal mortality in the moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and neonatal body mass was significantly lower than that in the mieroalbuminuria group (P ＜ 0.05).The incidences of FGR,premature birth and poor neonatal resuscitation and perinatal mortality in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the neonatal body mass was significantly lower than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in the neonatal asphyxia incidence between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and perinatal mortality in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the body mass of the newborns was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The incidence of neonatal asphyxia in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the incidences of FGR,premature birth,poor neonatal resuscitation,perinatal mortality and neonatal body mass between the two groups (P ＞ 0.05).Conclusions The of 24 h urinary protein quantitation is closely related to the pregnancy outcome in patients with pre-eclampsia,the 24 h urinary protein quantification should be regularly detected in the patients with pre-eclampsia.When the urinary protein quantitation is more than 2.0 g,the incidences of maternal complications and poor prognosis of the perinatal infants is significantly higher,but the boundary value of the 24 h urinary protein quantitation for the diagnosis of severe pre-eclampsia still needs further large sample study.

The mechanism of injury on the human glomerular endothelial cells (ciGENC) induced by preeclampsia serum was investigated. Concentration of maternal serum sFlt-1 protein was detected by ELISA. Fluorescently-labeled bovine serum albumin infiltrating through lower chamber of Transwell was measured by multifunction microplate reader. Morphologic change of ciGENC was observed under inverted phase contrast microscope. The concentration of sflt-1 in preeclampsia groups was significantly increased as compared with control group (P<0.01). Permeability in preeclampsia groups was significantly increased as compared with control group (P<0.01). By contrast with severe preeclampsia group, the permeability of ciGENC monolayer in mild preeclampsia group was decreased significantly (P<0.05). Intervention of exogenous VEGF significantly decreased permeability of ciGENC in preeclampsia groups. It was concluded that sFlt-1 increased ciGENC permeability by damaging integrity of endothelial barrier function.