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Objective To investigate the impacts of tanshinone ⅡA(Tan ⅡA)on lipopolysaccharide(LPS)induced proliferation and apoptosis of dental pulp stem cells by regulating the fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods Identification of human pulp stem cells(hDPSCs)isolated from the third molar of 18～20 years old patients requiring orthodontics.Lps-induced hDPSCs were treated with low,medium and high doses of Tan ⅡA,and then human recombinant FasL protein(rh FasL)was used to intervene the LPS-induced hDPSCs after high dose Tan ⅡA.Proliferation and apoptosis of hDPSCs,levels of tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 in hDPSCs supernatant,proliferating cell nuclear antigen(PCNA),Cleaved aspartate-specific cysteine proteinase-3(Cleaved Caspase-3),Fas,FasL protein expression were detected.Results hDPSCs were successfully isolated.In a dose-dependent manner,Tan ⅡA promoted LPS-induced proliferation,inhibited apoptosis,up-regulated PCNA protein expression,and inhibited TNF-α,IL-6 level,Cleaved Caspase-3,Fas,and FasL protein expression.The effect of rh FasL on LPS-induced dental pulp stem cells was opposite to the above indexes(P<0.05).rh FasL attenuates the effect of high-dose Tan ⅡA on pro-liferation and apoptosis of LPS-induced dental pulp stem cells.Conclusion Tan ⅡA may promote LPS induced hDPSCs proliferation and inhibit apoptosis by inhibiting the Fas/FasL signaling pathway.
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Objective To investigate the relationship between the levels of serum glycosylated hemoglobin (HbA1c), serum visfatin, glucose and lipid metabolism and the perinatal outcome in pregnant women with normal glucose tolerance (NGT). Methods A total of 135 NGT pregnant women diagnosed by oral glucose tolerance test (OGTT) entered this study. Patients were divided into two groups, high HbA1c group (HbA1c≥5.5%, n=66) and normal HbA1c group (HbA1c＜5.5%, n=69). The clinical data were collected in two groups. HbA1c levels were measured by HPLC, and the serum visfatin levels were measured by ELISA method. The values of body mass index (BMI), pregnant weight gain, visfatin, glucose and lipid met-abolic levels were analyzed between two groups. The relationship between levels of HbA1c, visfatin, glucose and lipid metab-olism and perinatal outcome were compared between two groups.Results The values of mean pregnancy weight gain, visfa-tin, total cholesterol (TC), triacylglycerol (TG), low density lipoprotein (LDL) and high density lipoprotein (HDL) were higher in high HbA1c group than those of normal HbA1c group. In NGT women with high HbA1c level,there were higher average birth weight, lower blood glucose level in newborn babies and higher rates of cesarean section. The incidences of polyhydram-nios, meconium-stained fluid, macrosomia and neonatal hypoglycemia were also higher in NGT women with high HbA 1c than those of normal HbA1c group. There was a positive relationship between HbA1c level, pregnant weight gain and visfatin and birth weight of newborn babies in 135 NGT pregnant women. Conclusion The excessive weight gain during pregnancy can induce the up-regulation of the visfatin level, resulting in the increased blood glucose level. The optimal timing and mode of delivery could prevent the occurrence of adverse pregnancy outcomes.
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Objective To investigate the expression of transforming growth factor-β1 (TGF-β1),endoglin (Eng) and their mRNA in placenta and superficial myometrium of patients with gestational hypertension or preeclampsia and to explore the role of Eng and TGF-β1 in the pathogenesis of gestational hypertension or preeclampsia.Methods One hundred and ten pregnant women were selected in the Second Affiliated Hospital of Tianjin Medical University from April 2009 to April 2010 who underwent cesarean sections and were divided into four groups:gestational hypertension group (n=30),mild preeclampsia group (n=30),severe preeclampsia group (n=30) and control group (normal pregnant women without labor and perinatal complications,n=20).The tissues of placenta and superficial myometrium were collected during cesarean section.Protein levels of Eng and TGF-β1 were detected by Western Blot.Real-time fluorescence reverse transcription polymerase chain reaction was used to detect the Eng and TGF-β1 mRNA expression.One-way ANOVA was used to compare among groups,Student-Newman-Keuls test was used to compare the differences between groups; relation between groups was analyzed by Pearson relation and linear regression.Results In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng mRNA level was 1.00,1.27±0.58,1.54±0.41 and 1.83±0.35,and the TGF-β1 mRNA level was 1.00,1.64 ± 0.33,1.92± 0.38 and 2.23 ± 0.53 in placenta respectively; those figures changed to 1.00,1.32±0.46,1.59±0.37 and 1.93±0.52,and 1.00,1.71 ± 0.45,1.91 ± 0.51 and 2.37 ± 0.46 in superficial myometrium respectively.The Eng and TGF-β1 mRNA levels of control group were lower than those of the other three groups (P＜0.05).The higher the mRNA level,the more severe the disease (P＜ 0.05).In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng protein expression was 0.11±0.07,0.15± 0.05,0.18 ± 0.06 and 0.43 ± 0.04,and the TGF-β1 protein expression was 0.11 ±0.02,0.26 ± 0.05,0.27± 0.03 and 0.88 ± 0.09 in placenta respectively; those figures changed to 0.14±0.06,0.16±0.04,0.20±0.08 and 0.46±0.05,and 0.15±0.03,0.29±0.06,0.31±0.04 and 0.91 ±0.08 in superficial myometrium respectively.The Eng and TGF-β1 protein levels of control group were lower than those of the other three groups (P＜0.05).The higher the protein level,the more severe the disease (P＜0.05).In the gestational hypertension group,mild and severe preeclampsia group,there were positive correlations between Eng and TGF-β1 protein levels in placenta (r=0.57,0.61 and 0.60 respectively,P＜0.05) and superficial myometrium (r=0.59,0.62 and 0.61 respectively,P ＜ 0.05).Conclusions Eng and TGF-β1 might play a role in pathogenesis of gestational hypertension and preeclampsia.
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Objective To investigate the effects of soluble endoglin(sEng)on nitric oxide (NO)production and endothelial nitric oxide synthase(eNOS)phosphorylation in cultured human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells within 3 passages seeded in culture plates of 96 wells,were stimulated by total culture medium(control group)or sEng (1,10 and 100 μg/L)respectively.Cells and medium were collected after cells were cultured for 6,12 and 24 hours respectively.The concentration of the metabolites of NO in each group was measured by nitrate reductase method.The expression of eNOS and eNOS-Ser(p)1177 were detected by Western blot.The expression of eNOS mRNA in each group was detected by real-time fluorescence reverse transcription-polymerase chain reaction.Analysis of variance,LSD method and pearson correlation were used to compare the difference between groups.Results(1)The concentration of the metabolites of NO in 1,10 and 100μg/L sEng groups was(59.25±1.63),(41.08±2.71)and (30.38±1.63)μmol/L respectively after cultured for 6 hours;(54.98±3.34),(35.00±8.60)and (19.82±3.75)μmol/L for 12 hours; and(46.14±4.93),(30.24±2.08)and(12.78±5.01)μmol/L for 24 hours.There was no significant changes in control group with time going by(F=2.30,P=0.14).The concentration of the metabolites of NO was significantly lower in sEng group,and which had negative correlation with culture time(r=-0.98,P＜0.05)and dose(r=-0.88,P＜0.05).(2)The expression of eNOS in 1,10,100 μg/L sEng groups was 0.71 ± 0.00,0.47 ± 0.00 and 0.32±0.00 after cultured for 6 hours; 0.58±0.00,0.42±0.00 and 0.25±0.00 for 12 hours; and 0.49±0.00,0.33±0.00 and 0.18±0.00 for 24 hours.While the expression of eNOS and eNOS-Ser (p)1177/eNOS had no significant changes in control group with time going by(F=3.59 and 0.37,P=0.09 and 0.80).The expression of eNOS protein and eNOS-Ser(p)1177 decreased significantly in sEng groups,which had negative correlation with culture time(r=0.98 and-0.96,P＜0.05)and dose(r=-0.76 and-0.79,P＜0.05).(3)The expression of eNOS mRNA decreased significantly in sEng groups.Which also had negative correlation with culture time(r=-0.51,P＜0.05)and dose(r=-0.82,P＜0.05).Conclusions sEng might inhibit eNOS activity by blocking 1177 Ser phosphorylation to decrease NO production.
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Objective To explore the ralationship between maternal serum level of soluble endoglin (sEng) in advanced gestations and hypertensive disorders comlicating pregnaney(HDCP). Methods The serum levels of sEng were analyzed using enzyme-linked immunosorbent assay (ELISA). Blood samples were obtained from 62 pregnant women with HDCP at 35-39 weeks' gestation (20 gestational hypertension, 20 mild preeclampsia, 19 severe preeclampsia and 3 eclampsia), and 20 normal pregnant women at 37-39 weeks' gestation (control). Results The serum sEng levels in normal, gestational hypertension, mild preeclampsia, severe preeclampsia and eclampsia group were (6.24±0. 26) ng/ml; (6. 56±0. 29) ng/ml; (7.47±0. 31) ng/ml; (8. 71± 0. 37) ng/ml and (9.69±0. 28) ng/ml, respectively. The serum sEng levels in the preeclampsia and eclampsia group were significantly higher than those in the gestational hypertension and normal group (P<0. 01), that of the severe preeclampsia group was significantly higher than the mild preeclampsia (P<0. 01), and that of the eclampsia group was significantly higher than the preeclampsia group (P<0. 01). However, no difference was found between the gestational hypertension and normal group (P>0. 05). Conclusions The increased serum level of sEng may participate in the genesis of HDCP.