Objective To analyse the effect of ursodeoxycholic acid on serum levels of cholyglycine ( CG ) , conjugated bile acid ( CBA ) and soluble vascular cell adhesion molecules (sVCAM-1) in patients with intrahepatic cholestasis of pregnancy.Methods 56 patients who were diagnosed with intrahepatic cholestasis of pregnancy in our hospital were collected.All patients were randomly divided into experimental group and control group, 28 cases in each group. The control group were treated with dexamethasone, and the experimental group were treated with the treatment of ursodeoxycholic acid, after 7d of treatment, the serum levels of glucocholic acid , CBA, ALT, AST and sVCAM-1 were detected in all patients. Results After treatment, compared with control group, the serum CG,TBA,ALT,AST and sVCAM-1 levels were significantly lower in the experimental group,and the difference was statistically significant (P<0.05).Conclusion The ursodeoxycholic acid can significantly reduce the serum CG,TBA, ALT,AST and sVCAM-1 levels in patients with intrahepatic cholestasis of pregnancy,improve pregnancy outcome,with guidance significance for clinic.

This study examined in vitro effect of lipoxin A(4) (LXA(4)) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA(4) at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca(2+)](i) of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca(2+)](i) of monocytes in the PE group were significantly higher than those in the control group. LXA(4) significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA(4), in the PE group, the [Ca(2+)](i) concentration of monocytes was significantly reduced. It was concluded that LXA(4) may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.

Objective To improve the knowledge of paediatric pulmonary arterial hypertension(PAH) and to elevate the level of early diagnosis and treatment.Methods The clinical data of 45 PAH patients admitted in Shanghai Children's Hospital from Jan.2006 to Dec.2012 were reviewed,including clinical manifestation,laboratory examination,diagnosis,treatment and prognosis.Results Of the 45 PAH patients,21 cases (46.7％) were male and 24 cases (53.3 ％) were female,with an average age of 2.5 years old.Among them idiopathic PAH was in 24 cases (53.3 ％) and secondary PAH was in 21 cases [including interstitial lung disease in 4 cases,upper airway obstruction in 3 cases,systemic lupus erythematosus in 3 cases,hepatic disease in 3 cases,including hepatic hemangioma 1 case,liver cirrhosis portal hypertension 1 case and autoimmune hepatic sarcoidosis 1 case,tachycardia induced cardiomyopathy in 2 cases,extensive pulmonary small artery stenosis in 2 cases,human immunodeficiency virus (HIV) infection in 1 case,hypothyroidism in 1 case,and familial PAH in 2 cases].Main clinical manifestations were anhelation after exercise (71.1％),fatigue (68.9％),cough (48.9％),chest tightness (26.7％),chest pain (33.3％),syncope (8.9％),et al.The most common physical signs were splitting of the second sound in pulmonary valve area (93.3％),followed by tricuspid murmur (77.8％),cyanosis (53.3％),hepatomegaly (42.2％),hydropericardium (28.9％),and oedema (11.1％),et al.Twenty-seven cases did cardiac catheterization,compared with idiopathic PAH and secondary PAH,pulmonary artery systolic pressure,mean pulmonary artery pressure,pulmonary capillary wedge pressure,pulmonary arteriolar resistance index had significant difference(P ＜ 0.05).Thirty-one cases' condition improved after treatment,11 cases without any improvement and 3 cases died during follow-up visit.Conclusions PAH is a rare disease with no specific symptom and can be easily misdiagnosed in children.Ultrasonic cardiogram and cardiac catheterization are helpful in diagnosis.Actively looking for the cause can improve the prognosis.

Objective To investigate the effect of lipoxin A4(LXA4) on interleukin-1β(IL-1β)production of monocytes in maternal peripheral blood from severe preeelampsia women and its relationship with cytosolic free calcium([Ca2+]i)concentration. Methods Peripheral venous blood was drawn from 15 women with severe preeelampsia(SPE group)and 20 normal pregnant women (control group)who were admitted to the First Affiliated Hospital of Wenzhou Medical College from October 2008 to May 2009.Monoeytes were obtained from peripheral blood with Ficolt density gradient centrifugation and then were suspended in RPMI-1640 culture supplemented with LXA4 at the final concentrations of 0,10 and 100 nmol/L,respectively.IL-1β levels in monocytes supernatant were detected by enzyme-linked immunosorbent assay.The cytoplasma[Ca2+]i of cultured monocytes and its variations affected by LXA4 were measured by laser scanning confocal microscope. Results (1) After incubation with different concentrations of LXA4(0,10,100 nmol/L)for 24 h,the levels of IL-1β in SPE group were(63.16±8.20)pg/L,(53.71±8.08)pg/L and(43.16±6.89)pg/L,respectively, indicating a significant inhibition effect on IL-1β level in a dose-dependent manner (P<0. 05). The IL-1β levels in the control group were (19.22±7.43) pg/L, (16.99±6.32) pg/L and (15. 18±5.24) pg/L, correspondingly (P>0.05). (2) Without LXA4, the [Ca2+ ]i concentrations of monocytes in the SPE group were higher than that of the control (1028.09 ± 160. 52 vs 323.61 ±87.86, P<0. 05). After treatment with 100 nmol/L LXA4, the [Ca2+ ]i concentration of monocytes significantly decreased in the SPE group (409.67± 116.73, P<0. 05). Conclusions In vitro LXA4 may inhibit the IL-1β production in monoeytes of SPE patients through decrease of the cytoplama [Ca2+ ]i concentration.

Objective To investigate the clinical characteristics and the optimal time of delivery in pregnant women with severe preeclampsia complicated with ascites. Methods A retrospective study was conducted on 179 severe preeclampsia mothers and their 195 neonates,presented in the First Affiliated Hospital of Wenzhou Medical College from Jan.2003 to Dec.2005,who were divided into two groups:32 complicated with ascites(ascites group)and 147 without(non-ascites group). The general conditions,mode of delivery and complications including eclampsia,hemolysis,elevated serum level of 1iver enzymes,and low platelets(HELLP syndrome),liver failure,renal failure,heart failure,hypoproteinemia,placental abruption,postpartum hemorrhage and puerperal infection,were also analyzed.Clinical data of all infants(38 from ascites group and 157 from non-ascites group)were analyzed.The incidence and mortality rate of small for gestational age(SGA)in both group within the same gestational age group and those between different gestational age groups in the ascites group were compared. Results (1)The average gestations at admission and delivery in the ascites group were earlier than the other[admission:(32.5±2.1)weeks vs(36.1±3.5)weeks;delivery:(34.1±2.3)weeks vs(37.2±1.5)weeks,P＜0.053.The rate of systemic antenatal care in the ascites group waslowcr than that of the non-ascites group(25.0%vs 53.7%,P＜0.05).More complications werefound in the ascites group than in the non-ascites group(hypoproteinemia:100.0%vs 47.0%;liver and renal failure:31.2%vs 8.2%;HELLP syndrome:9.4%vs 2.0%;postpartum hemorrhage:18.8%vs 2.0%;all P＜0.05).(2)The incidence of SGA in the ascites group was all higher than that in the non-ascites group,however,significant differences was only found between the tWO groups at＞36 weeks(7/9 vs 30.2%,P＜20.05).The perinatal mortalily rates of SGA in the ascites group at＜32 weeks and 32～34 weeks were significantly higher than that in the non-aseites group respectively(＜32 weeks:69.2%vs 19.2%,P＜0.05;32～34 weeks:2/7 vs 0,P＜0.05).(3)The highest perinatal mortality rate and the highest incidence of SGA in the ascites group were found in the groups of＜232 weeks and＞36 weeks,respectively. Conclusions The early onset of ascites and higher rate of complications in severe preeelamptie women implies the adverse maternaI and fetal outcomes.Ascites in severe preeclampsia cases should alert the clinicians.The optimal time for delivery might be at 32～36 weeks of gestations.

Objective To investigate protective effects and mechanisms of lipoxinA4(LXA4)on human umbilical vein endothelial cells(HUVEC)under hypoxia in vitro.Methods The HUVEC culture were divided into groups as followed:added M199 cudture medium as normal contml groups,added CoCl2 to mimic hypoxia in vitro as hypoxia group and added different concentrations of LXA4(1,10,100 nmol/L)were added to the induced hypoxiai HUVEC as agents intervention group.Morphological changes of HUVEC were observed by using inverted phase contrast mieroscope.The influence of LXA4 on cell survival was investigated by methyl thiazolyl tetrazolium(MTT) assaying method after the treatment with different concentrations of LXA4 and 100 nmol/L lipoxinA4 according to different time(4,8,12 and 24 hours).The expression of von-willebrand factor (vWF) was detected by immunocytoehemistry method. The changes of cytosolic Ca2+ were measured by laser scanning confocal microscope. Results ( 1 ) Morphological changes:the cells under hypoxia lost its normal shapes and showed necrosis, while the cells cocuhured with 100 nmol/L LXA4 were normal appropriately. (2)Survival rate: the survival rates of HUVEC under hypoxia was (40. 1±3.9) % and increased to ( 52. 9 ± 1.4) %, (64. 1 ± 3. 3 ) %, ( 76. 6 ± 1.6) % respectively when added with LXA4 with concentration of 1,10, 100 nmol/L into culture medium. There was significant different survival rate when compared with that of hypoxia group. (3) The level of vWF: The expression of vWF was decreased with the increasing concentrations of LXA4 added into culture medium, the gray values were 203.9 ±0. 7 in 1 nmol/L,204.6 ±0. 9 in 10 nmoL/L,191.8 ±0. 5 in 100 nmol/L respectively, which reached statistical difference in comparison with that of hypoxia groups (P<0. 05). (4) Confocai analysis:the intracellular free Ca2+ concentrations of HUVEC were intensified with LXA4 treatment. Conclusions LXA4 plays an important role in keeping the normal shape of HUVEC under hypoxia, can enhance survival of hypoxial HUVEC and decrease the level of vWF in cytoplasm. The protective mechanism might be via decreasing mitochondria Ca2+ overload and increasing cytoplasm Ca3+ by nucleus Ca2+ transference.

Objective To explore lipoxinA4 (LXA4) expression in maternal serum of pregnant women and the protective effect and mechanism of LXA4 on trophoblastic cells from oxidative injury. Methods Trophoblastic cells were randomized into six groups: Control group; Lipopolysaccharides (LPS) group, cells were stimulated by 10 μg/ml LPS for 24 h; Intervention group, cells stimulated by LPS were treated with 100 nmol/L LXA4 for 24 h; LXA4 group, cells were treated with 100 nmol/L LXA4 for 24 h; Antagonistic group, cells stimulated by LPS were treated with 100 nmol/L LXA4 plus 100 μmol/L N-tert-butoxycarbonyl-2-pyrrolidine (BOC-2) for 24 h; BOC-2 group, trophoblastic cells stimulated by LPS were treated with 100 μmol/L BOC-2 for 24 h. The serum concentration of LXA4 in normal group and preeclampsia group was detected by ELISA. The intracellular formation of reactive oxygen species (ROS) was detected by 2,7-dichlorofluorescein diacetate (DCFH-DA) as a fluorescent probe. SOD mRNA was analyzed by RT-PCR. SOD and Nrf2 protein expressions were analyzed by Western blot. The levels of SOD in trophoblastic cells were detected by using detection kit. Results (1) The serum concentration of LXA4 was significantly lower in preeclampsia group (165.53±18.89) pg/L than in the control [(545.67±30.91) pg/L, P<0.01]. (2) Compared with control group, the levels of ROS in LPS group were significantly higher, DCF density of trophoblastic cells increased from 53.00±3.08 to 77.00±5.83 (P<0.01). The expression of nuclear Nrf2 protein, SOD mRNA and protein in LPS group were obviously decreased (P<0.01). The levels of SOD in LPS group were also significantly lower (P<0.01). (3) Compared with LPS group, the levels of ROS in intervention group were significantly lower, DCF density of trophoblastic cells decreased from 77.00±5.83 to 62.00±3.39 (P<0.01). The expression of nuclear Nrf2 protein, SOD mRNA in intervention group were obviously increased (P<0.01), so did the SOD protein expression (P<0.05). The levels of SOD were significantly increased from (4.77±0.34) U/ml to (8.93±0.53) U/ml (P<0.01). (4) The levels of SOD in antagonistic group were lower than in intervention group, but still higher than LPS group. [(6.23±0.41) U/ml vs (8.93±0.53) U/ml (P<0.01) or (4.77±0.34) U/ml (P<0.01)]. No significant difference was found in the levels of SOD between BOC-2 and LPS group (P>0.05). Conclusions LXA4 can significantly reduce the oxidative stress of placental trophoblastic cells stimulated by LPS. LXA4 can bind to lipoxin receptors and activate Nrf2-ARE signaling pathway playing a protective effect. So LXA4 in pregnant women can affect the oxidative stress of placenta.

ObjectiveTo investigate the effects of 5(S),6(R),7-trihydroxyheptanoic acid methyl ester (BML-111) on pregnant mice with fetal growth restriction(FGR) induced by antenatal dexamethasone and its probable mechanism. MethodsThe mice were mated overnight,with day 1 of pregnancy designated as the day on which spermatozoa were presented in a vaginal smear.The pregnant mice were then randomly divided into control group,dexamethasone group and BML-111 group.From 9 to 14 days of pregnancy,pregnant ICR mice of control,dexamethasone and BML-111 group were treated separately with saline,dexamethasone(5 mg/kg) and dexamethasone at 8:00 am,and two hours later they were treated separately again with 1 mg/kg saline,saline and BML-111.On the day 18 of gestation,they were sacrificed after blood were collected from their eyeballs.The serum lipoxin A4 was measured with enzyme-linked immunosorbent assay. Fetuses were delivered by cesarean section; the placenta and uterus were immediately removed and frozen.Gene expressions of 11β-hydroxysteroid dehydrogenase 2 ( 11β-HSD2 ),interleukin-1β (IL-1β) in placenta and lipoxin A4 receptor-formyl peptide receptor 3 (FPR3)in uterine were detected by reverse transcriptionpolymerase chain reaction and compared with analysis of variance.The 11β-HSD2 protein in mice placenta was detected by immunohistochemistry. ResultsThe mean fetal weight of dexamethasone group was (0.823±0.054) g,lower than that of the control group and BML-111 group [(1.103±0.218) g and (0.992 ± 0.207) g] (t =- 4.108 and - 2.890,P ＜ 0.05 respectively).Protein expression of 11β-HSD2 in dexamethasone group (0.030±0.019) was weaker than that in control group (0.058±0.015,t=-3.107,P＜0.05) or in BML-111 group (0.049±0.026,t=-2.211,P＜0.05).The expression of 11β-HSD2 mRNA in dexamethasone group (0.457±0.062) was lower than that in control group (0.943±0.057,t=-9.418,P＜0.05) or in BML-111 group (0.698±0.071,t=-4.617,P＜0.05).Expression of IL-1β mRNA in dexamethasone group (0.543±0.103)was less than that in control group (0.710± 0.085,t=-3.736,P＜0.05) but more than that in BML-111 group (0.229 ±0.031,t=7.025,P＜0.05). The expression of FPR3 mRNA in dexamethasone group (0.323 ± 0.019) was less than that in control group (0.857 ± 0.057,t =-14.630,P＜0.05) or in BML-111 group (0.499 ±0.050,t=-4.822,P＜0.05).The serum concentration of lipoxin A4 in dexamethasone group was lower than that in control group [(64.463±22.144) pg/ml vs (101.610±24.916) pg/ml,t=3.152,P＜0.05].ConclusionsBML-111 regulate the expression of 11β-HSD2 and then protect against FGR resulted from too much prenatal application of dexamethasone.

Objective To investigate the expression and clinical significance of lipoxin A4,leukotrienc C4,lipoxygenase-5 in peripheral blood of pregnant women with different types of severe preeclampsia.Methods Forty-five singleton pregnant women who accepted antenatal care and delivered in First Affiliated Hospital of Wenzhou Medical College were enrolled in this study from December 2010 to June 2011.All objects were divided into normal pregnancy group (n=20),early onset severe preeclampsia group (n=10) and late onset severe preeclampsia group (n=15).Enzymelinked immunosorbent assay was used to detect lipoxin A4 and leukotriene C4 levels in peripheral blood.The level of lipoxygenase-5 mRNA in white blood cells was detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction.The differences of lipoxin A4,leukotriene C4 and lipoxygenase-5 mRNA among groups were compared by analysis of variance and LSD-t test;and correlations among their expressions were analyzed by linear regression.Results Lipoxin A4 level in early and late onset severe preeclampsia group was (355.3±116.0) pg/ml and (389.7±117.5) pg/ml,which were both significantly lower than that in normal pregnancy group [(555.0±139.8) pg/ml] (t=-4.03 and-3.77,P＜0.05 respectively).The leukotriene C4 level in early and lateonset severe preeclampsia and normal pregnaney group was (591.3±185.5) pg/ml,(510.3±197.1) pg/ml and (496.9 ± 158.8) pg/ml,no statistical difference were found (F=0.889,P＞0.05) ; neither did the expression of lipoxygenase 5 mRNA,which was 4.2± 1.9 in normal pregnancy group,4.8 ± 2.0 in early onset severe preeclampsia group and 4.4 ± 1.2 in late onset severe preeclampsia group (F=0.311,P＞0.05).There was no correlation among the levels of lipoxin A4,leukotriene C4 and lipoxygenase-5 mRNA in each group (P ＞ 0.05).Conclusions Early and remarkable decreasing of lipoxin A4 level might contribute to the development of early onset severe preeclampsia.