Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

Objective To explore the feasibility and safety of vaginal delivery with scarred uterus after cesarean section.Methods Sixty two parturients after cesarean section undergoing trial vaginal delivery in our department from October 2016 to October 2017 were enrolled in study (study group);60 primiparous women admitted at the same period were taken as the control group.The successful rate of vaginal delivery,the duration of labor and the incidence of postpartum complications were analyzed.The postpartum depression was assessed with Edinburgh Postpartum Depression Scale 6 weeks after delivery and compared between two groups.Results The vaginal delivery was successful in 49 parturients of each group with a successful rate of 79.0％ (49/62) and 81.7％ (49/60) respectively (x2=0.13,P=0.71).The first and the second stages of labor in the study group were (364± 105)min and (54±31)min,respectively,which were shorter than those of the control group [(388±93)min and (63± 18)min,t=2.325,P=0.03;t=2.145,P=0.04].The incidence rates of postpartum hemorrhage,infection and urinary retention were 3.2％ (2/62) and 1.7％ (1/60),1.6％ (1/62) and 1.7％ (1/60),4.8％ (3/62) and 3.3％ (2/60) in study group and control group,respectively (x2=0.309,0.001 and 0.176,P＞0.05).The Apgar scores of the newborns in two groups were 8.7± 2.1 and 8.5± 1.8 (t=1.415,P=0.14) and the postpartum depression scores of the mothers were 13.7±4.3 and 12.4±3.2 (t=1.203,P=0.33),respectively.Conclusion Vaginal delivery can be chosen in parturients with scarred uterus after cesarean section,if preparations are adequate,indications of vaginal delivery are sufficient and the course of labor is closely monitored.

Objective To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression.Methods The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured.The cultured cells were treated with (1) forskolin groups: different concentration (0,2.5,5,50 or 100 μmol/L) of forskolin treated cells for 2 hours,and the optimal concentration of forskolin treated cells with different time (0,1,2,10 or 20 hours) ; (2)SP-cAMP groups: different concentration (0,2.5,5,50 or 100 μmol/L) of SP-cAMP treated cells for 2 hours,and the optimal concentration of SP-cAMP treated cells with different time (0,1,2,10 or 20 hours); (3)H-89 groups: different concentration (0,5,10,50 or 100 μmol/L) of H-89 treated cells for 2 hours,and the optimal concentration of H-89 treated cells with different time (0,1,2,10 or 20 hours).The level of intracellular cAMP and activity of PKA were detected by using ELISA,and immunohistochemistry was used to detect the localization of AQP3,the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis.Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay.Results (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group.(2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin,SP-cAMP and H-89 treatment (P ＞ 0.05).(3) After different concentration of forskolin treated 2 hours,the expression of total CREB had no significant difference among them(P ＞ 0.05).While the expression of cAMP level,PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 2.5 μmol/L,5 μmol/L,50 μmol/L forskolin group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 5 μmol/L forskolin group were higher than that in 2.5 μmol/L and 50 μmol/L (P ＜ 0.05).The optimal forskolin concentration was 5 μmol/L.(4) After different concentration of SP-cAMP treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 5 μμmol/L,50 μmol/L SP-cAMP group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 50 μmol/L SP-cAMP group were higher than that in 5 μmol/L (P ＜0.05).The optimal SP-cAMP concentration was 50 μmol/L (5) After different concentration of H-89 treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were lower in 10 μmol/L,50 μmol/L and 100 μmol/L H-89 group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 10 μmol/L H-89 group were lower than that in 50 μmol/L,100 μmol/L (P ＜ 0.05).The optimal H-89 concentration was 10 μmol/L.(6) p-CREB and AQP3 protein expression were significantly lower in 5 μmol/L forskolin combined 10 μmol/L H-89 incubating 2 hours group when compared with 5 μmol/L forskolin,but higher than that in 10 μmol/L H-89 treated group (P ＜ 0.05).Total CREB was no significant difference among the three groups (P ＞ 0.05).Conclusion cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

OBJECTIVETo study the association between phthalate esters (PAEs) metabolites in maternal urine and 11beta-hydroxysteroid dehydrogenase type 2 (11β-HSD2 ) enzyme activity, explore the possible mechanism of PAEs effect on fetal development.

METHODSAll of 33 cases of intrauterine growth retardation (IUGR) newborn were selected by random sampling in 2012. And 33 cases of normal control newborn were enrolled, use high performance liquid chromatography-tandem mass spectrometry method was used to detect 4 kinds of phthalate esters (PAEs) metabolites in maternal urine: mono-n-butyl phthalate ester (MBP), mono (2-ethylhexyl) phthalate (MEHP), mono (2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono (2-ethyl-5-oxohexyl) phthalate (MEOHP) and three kinds of cortisol corticosterone metabolites, tetrahydrocortisol (THF), allo-tetrahydrocortisol (allo-THF), tetrahydrocortisone (THE), and analyze the association between phthalate esters (PAEs) metabolites in maternal urine and 11β-HSD2 enzyme activity.

RESULTSMBP, MEHP, MEHHP, MEOHP metabolites can be detected in 98% (65 cases) , 89% (59 cases), 91% (60 cases), 91% (60 cases) of all 66 maternal urine samples, respectively. The median concentrations of test material in case group were 31.20 ng/ml for MBP, 24.61 ng/ml for MEHHP, 11.72 ng/ml for MEOHP and 48.67 ng/ml for SumDEHP which were significantly higher than those of the control group (were 17.32, 12.03, 5.68 and 28.64 ng/ml); 11β-HSD2 activity in case group ((THF+allo-THF)/THE = (0.79 ± 0.09) ng/ml) was significantly lower than that of the control group((THF+allo-THF)/THE = (0.58 ± 0.04) ng/ml); PAEs metabolites MBP (β' = 1.12), MEHHP(β' = 1.14), MEOHP(β' = 1.10), SumDEHP(β' = 1.08) in baby boy mother's urine was reversely correlated to 11β-HSD2 activity.

CONCLUSIONSPAEs could affect fetal development by inhibit 11β-HSD2 activity.

11-beta-Hydroxysteroid Dehydrogenase Type 2 ; Chromatography, Liquid ; Diethylhexyl Phthalate ; analogs & derivatives ; Fetal Development ; Humans ; Infant, Newborn ; Male ; Mass Spectrometry ; Phthalic Acids ; Tetrahydrocortisol ; analogs & derivatives ; Tetrahydrocortisone

Objective To study the association between phthalate esters ( PAEs ) metabolites in maternal urine and 11beta-hydroxysteroid dehydrogenase type 2(11β-HSD2 ) enzyme activity, explore the possible mechanism of PAEs effect on fetal development.Methods All of 33 cases of intrauterine growth retardation ( IUGR) newborn were selected by random sampling in 2012.And 33 cases of normal control newborn were enrolled , use high performance liquid chromatography-tandem mass spectrometry method was used to detect 4 kinds of phthalate esters ( PAEs) metabolites in maternal urine:mono-n-butyl phthalate ester (MBP), mono (2-ethylhexyl) phthalate (MEHP), mono (2-ethyl-5-hydroxyhexyl) phthalate(MEHHP), mono ( 2-ethyl-5-oxohexyl ) phthalate ( MEOHP ) and three kinds of cortisol corticosterone metabolites , tetrahydrocortisol (THF), allo-tetrahydrocortisol (allo-THF), tetrahydrocortisone (THE), and analyze the association between phthalate esters ( PAEs) metabolites in maternal urine and 11β-HSD2 enzyme activity.Results MBP,MEHP,MEHHP,MEOHP metabolites can be detected in 98%(65 cases),89%(59 cases), 91%(60 cases),91%(60 cases)of all 66 maternal urine samples, respectively.The median concentrations of test material in case group were 31.20 ng/ml for MBP, 24.61 ng/ml for MEHHP, 11.72 ng/ml for MEOHP and 48.67 ng/ml for SumDEHP which were significantly higher than those of the control group ( were 17.32,12.03,5.68 and 28.64 ng/ml); 11β-HSD2 activity in case group (( THF +allo-THF)/THE =(0.79 ±0.09) ng/ml) was significantly lower than that of the control group (( THF+allo-THF)/THE=(0.58 ±0.04) ng/ml);PAEs metabolites MBP(β′=1.12),MEHHP(β′=1.14),MEOHP(β′=1.10), SumDEHP (β′=1.08 ) in baby boy mother′s urine was reversely correlated to 11β-HSD2 activity.Conclusions PAEs could affect fetal development by inhibit 11β-HSD2 activity.

Objective To study the association between phthalate esters ( PAEs ) metabolites in maternal urine and 11beta-hydroxysteroid dehydrogenase type 2(11β-HSD2 ) enzyme activity, explore the possible mechanism of PAEs effect on fetal development.Methods All of 33 cases of intrauterine growth retardation ( IUGR) newborn were selected by random sampling in 2012.And 33 cases of normal control newborn were enrolled , use high performance liquid chromatography-tandem mass spectrometry method was used to detect 4 kinds of phthalate esters ( PAEs) metabolites in maternal urine:mono-n-butyl phthalate ester (MBP), mono (2-ethylhexyl) phthalate (MEHP), mono (2-ethyl-5-hydroxyhexyl) phthalate(MEHHP), mono ( 2-ethyl-5-oxohexyl ) phthalate ( MEOHP ) and three kinds of cortisol corticosterone metabolites , tetrahydrocortisol (THF), allo-tetrahydrocortisol (allo-THF), tetrahydrocortisone (THE), and analyze the association between phthalate esters ( PAEs) metabolites in maternal urine and 11β-HSD2 enzyme activity.Results MBP,MEHP,MEHHP,MEOHP metabolites can be detected in 98%(65 cases),89%(59 cases), 91%(60 cases),91%(60 cases)of all 66 maternal urine samples, respectively.The median concentrations of test material in case group were 31.20 ng/ml for MBP, 24.61 ng/ml for MEHHP, 11.72 ng/ml for MEOHP and 48.67 ng/ml for SumDEHP which were significantly higher than those of the control group ( were 17.32,12.03,5.68 and 28.64 ng/ml); 11β-HSD2 activity in case group (( THF +allo-THF)/THE =(0.79 ±0.09) ng/ml) was significantly lower than that of the control group (( THF+allo-THF)/THE=(0.58 ±0.04) ng/ml);PAEs metabolites MBP(β′=1.12),MEHHP(β′=1.14),MEOHP(β′=1.10), SumDEHP (β′=1.08 ) in baby boy mother′s urine was reversely correlated to 11β-HSD2 activity.Conclusions PAEs could affect fetal development by inhibit 11β-HSD2 activity.

Objective To investigate the role of mitogen-activated protein kinases (MAPK)/extracellular signal regulated kinase1/2 (ERK1/2) signal transduction pathway in regulating the expression of aquaporin 3 (AQP3) in human amnion epithelial cells.Methods Primary cell culture of human amnion epithelial cells deriving from amnion of term pregnancy with normal amniotic fluid volume (AFV) or isolated oligohydramnios was conducted in the Second Affiliated Hospital of Wenzhou Medical College from January to November 2011.Either group included 10 elective cesarean cases.The primarily cultured cells were treated with different concentrations (0,5,10,20 and 40 mol/L) of ERK inhibitors (U0126) for 12 h,and then the optimal concentration of U0126 which resulted in the lowest expression of phospho~ERK1/2 (p-ERK1/2) was added for different durations(0,2,6,12 and 24 h).Immunocytochemistry was used to detect the localization of AQP3 and Western blot analysis was used to examine the expression of total ERK1/2,p-ERK1/2 and AQP3 in human amnion epithelial cells.Statistical analysis was performed by t-test and one-way ANOVA.Results (1) Compared with those in normal AFV group,p-ERK1/2 and AQP3 expression in human amnion epithelium cells decreased in oligohydramnios group,respectively (p-ERK1/2:3.46 ± 0.33 and 2.46±0.25;AQP3:2.34 ± 0.18 and 1.56±0.10,t=9.243 and 13.292,P＜0.01).(2) In oligohydramnios groups,after treated with different concentrations of U0126,the expressions of total ERK1/2 did not change (F=0.365,P＞0.05).The expression of p-ERK1/2 and AQP3 in 5,10,20,40 μmol/L U0126 (p-ERK1/2:0.96±0.16,1.12±0.13,0.98±0.17 and 1.02±0.26; AQP3:1.10±0.09,1.12±0.08,1.13±0.06 and 1.11±0.06) were all significantly lower than those in 0 μmol/LU0126 group (p-ERK1/2:2.46±0.25; AQP3:1.56±0.10,P＜0.05).However,the expression of p-ERK1/2 and AQP3 showed no significant difference among 5,10,20,and 40μmol/L U0126 groups (P＞0.05).The optimal concentration of U0126 was 5 μmol/L.After treated with 5 μmol/L U0126 for 2 h,the expressions of p-ERK1/2 and AQP3 (1.27±0.29 and 1.44±0.12)were lower than those after treated for 0 h (2.55±0.12 and 2.15±0.09,P＜0.05).However,there was no significant difference among groups treated for 2,6,12 and 24 h.Therefore,the optimal treatment time was 2 h.(3) The expression of AQP3 was distributed in both cell membrane and cytoplasm in amnion epithelial cells with normal amniotic fluid volume or isolated oligohydramnios,but mainly in cytoplasm.U0126 did not change the localization of AQP3 expression.Conclusions U0126 inhibits the phosphorylation of ERK1/2 and expression of AQP3 of women with oligohydramnios,indicating that the MAPK/ERK1/2 signal transduction pathway might regulate the expression of AQP3 in human amnion epithelial cells,and therefore affect the balance of amniotic fluid volume.

Objective To investigate the iron status in the newborns of maternal gestational diabetes mellitus(GDM) women,and explore the mechanism of iron deficiency in these newborns.Methods From June 2008 to October 2011,64 GDM women (GDM group) and 71 healthy pregnant women (control group)who delivered in the Second Affiliated Hospital of Wenzhou Medical College and their newborns were studied prospectively.Serum ferritin (SF),serum transferrin receptor (sTfR),erythropoietin (Epo),haemoglobin (Hb),serum level of insulin and plasma glucose in cord blood was measured.The neonatal birth weight (BW) and birth weight Z Score(WAZ) was recorded.The concentrations of serum fasting insulin (FINS),fasting plasma glucose (FPG) and glycosylated hemoglobin (HbA1c)were tested for all the women before delivery.Results In the GDM group,the cord blood sTfR,Epo and serum level of insulin was (42 ± 10)nmol/L,(56 ±41) U/L and(18 ± 5) U/L,respectively.While in the control group,these were(35 ± 8)nmol/L,(41 ± 43) U/L and (10 ± 5) U/L,respectively.The cord blood sTfR,Epo and serum level of insulin in the GDM group were significantly higher than those in the control group (P ＜ 0.05).The cord blood SF in the GDM group[(60 ±36) μg/L] was significantly lower than that of the control group[(146 ±38) μg/L,P ＜ 0.01].The neonatal BW and WAZ in the GDM group [(3615 ± 538) g and 0.558] were significantly higher than those in the control group [(3449 ± 423) g and 0.224,P ＜ 0.05].No significant difference was found in the cord blood plasma glucose and Hb between the GDM group[(3.3 ± 1.0) mmol/L and (181 ± 18) g/L] and the control group [(3.0 ± 0.8) mmol/L and (176 ± 16) g/L,P ＞ 0.05].The FINS and HbA1c of the GDM group[(12.5 ±5.0) U/L and (6.5 ±0.7)％] were significantly higher than those in the control group [(10.9 ± 4.3) U/L and (5.3 ± 0.7) ％,P ＜ 0.05].The FPG of the GDM group and the control group were (5.3 ± 1.2) and (5.0 ± 1.0) mmol/L,respectively,with no statistically significant difference (P ＞ 0.05).Conclusion Maternal GDM may related to the iron deficiency of the newborns.

Objective To investigate factors and neonatal outcomes associated with histologic chorioamnionitis (HCA) in preterm premature rupture of membranes (PPROM).MethodsFrom Jan.2008 to Jun.2011,230 women with PPROM at 28 -33 +6 weeks of gestation undergoing deliveries in the Second Affiliated Hospital of Wenzhou Medical College were studied retrospectively.According to placental histopathologic findings,those patients were categorized into two groups,including 138 cases in histologic chorioamnionitis (HCA group ) and 65 cases in non-chorioamnionitis (control)group.Age,parity,gestational age of PPROM and delivery,latency period,oligohydramnios,white blood cell (WBC) count and serum C-reactive protein (CRP) level at admission and before delivery,the incidence of neonatal respiratory distress syndrome (NRDS),neonatal pneumonia,bronchopulmonary dysplasia,necrotizing enterocolitis,early-onset neonatal sepsis,abnormal brain sonography findings and mortality were compared between two groups.Results( 1 ) The incidence of HCA was 68.0.％ ( 138/203 ) in all 203 cases with PPROM.(2) The occurring ruptured membrane gestation in HCA group was ( 31.1 ± 1.5 ) weeks,which were significantly earlier than (32.0 ± 1.3 ) weeks in control group ( P ＜ 0.05 ).The level of CRP of (8.2 ± 14.9) mg/L before delivery in HCA group was significantly higher than (5.5 ±7.2) mg/L in control group (P ＜ 0.05).The rate of oligohydramnios and cesearean sections were 55.1％ (76/138) and 45.7％ (63/138) in HCA group,which were significantly higher than 30.8％ (20/65) and 29.2％ (19/65) in control group (P ＜0.05).There were no significant difference in patient's age,parity,WBC count and CRP at admission between two groups (P ＞ 0.05 ).The latency period did not show significant difference between (140± 116) hours in HCA group and (129 ± 125) hours in control group (P ＞ 0.05).(3) Using multivariable logistic regression models,oligohydramnios ( OR =2.937 ),gestational age of PPROM ＜ 32 weeks ( OR =2.352),serum CRP level ＞ 8 mg/L before delivery ( OR =4.923 ) and latency period ＞ 48 -168 hours (OR =4.439) were significantly associated with HCA (P ＜0.05).(4) The gestational age of delivery and birth weight of HCA group were significantly lower than those of control group [ ( 32.0 ± 1.5 ) weeks vs.( 32.7 ± 1.5 ) weeks,( 1680 ± 379) g vs.(2017 ± 333) g,respectively,P ＜ 0.05 ].The incidence of Apgar ＜7,abnormal brain sonograhy findings, neonatal pneumonia,bronchopulmonary dysplasia,early-onset neonatal sepsis and mortality in HCA group were significantly higher than those in control group [20.3％ (28/138) vs.7.7％ (5/65),14.5％ (20/138) vs.4.6％ (3/65),12.3％ (17/138) vs.3.1％(2/65),5.8％ (8/138) vs.0,6.5％ (9/138) vs.0,12.3％ (17/138) vs.3.1％ (2/65),respectively,P ＜ 0.05 ].The incidence of necrotizing enterocolitis ( 1.5％,2/138 ) in HCA group was higher than that of controlgroup(0) and the incidence of NRDS ( 18.8％,26/138) in HCA group did not show statistical difference with 21.4％ ( 14/65 ) in control group ( P ＞ 0.05 ).ConclusionsIt was found that HCA was significantly correlated with lower gestational age of PPROM,higher serum CRP level before delivery,prolonged latency period and oligohydramnios in PPROM.HCA could increase the neonatal morbidity and mortality.