Objective To investigate the relationship between placental endoglin expression and the pathogenesis of severe preeclampsia (SPE). Methods Forty nine pregnant women with SPE and 40 normal pregnant controls were collected in Peking University First Hospital from January 2006 to January 2008,among which,nine SPE patients complicated with fetal growth restriction (FGR),six with hemolysis,elevated liver enzymes and low platelets syndrome (HELLP syndrome) and 12 with heavy proteinuria. The expression of placental endoglin was detected and semi-quantified by immunohistochemistry.Data were analyzed by independent sample t test. Results Endoglin was presented on cell membranes of placental syncytiotrophoblasts and vascular endothelial cells. The endoglin density of SPE group was higher than that of control group (0.1621± 0.0029 vs 0.1576 ± 0.0038,t=- 6.367,P＜0.05).No significant difference in endoglin density was found between FGR group and non-FGR group (0.1611±0.0026 vs 0.1623±0.0029,t=1.107,P＞0.05) ; neither did the heavy proteinuria group and non-heavy proteinuria group (0.1611±0.0032 vs 0.1624±0.0027,t=1.350,P＞0.05).The endoglin density of HELLP group was lower than that of non-HELLP group (0.1595±0.0032 vs 0.1625±0.0027,t=2.495,P=0.016). Conclusions The elevated placental endoglin expression might contribute to the pathogenesis of SPE.

Objective To investigate the relationship between fasting plasma glucose (FPG) in early pregnancy and diagnosis of gestational diabetes mellitus (GDM) and to confirm the rationality of the new standard for GDM diagnosis in early pregnancy set by the International Association of Diabetes and Pregnancy Study Groups (IADPSG).Methods Clinical materials of 2761 pregnant women without diabetes mellitus,who accepted prenatal cares in Peking University First Hospital from April 1,2011 to December 31,2011,were collected and analyzed.The difference between FPG levels of GDM and non-GDM women was compared.According to the early pregnancy FPG level,the subjects were divided into group A (FPG＜5.1 mmol/L,n=2431) and B (FPG≥5.1 mmol/L,n=330).The incidence of GDM and pregnant outcomes of the two groups were compared with t or Chi-square test.Relationship between FPG and GDM was analyzed by Logistic regression and receiver operating characteristic curve.Results (1) Among the 2761 subjects,515 were diagnosed as GDM (18.7％) and the early pregnancy FPG level in GDM group was significantly higher than that in non-GDM group [(4.84±0.46) mmol/L vs (4.57 ± 0.35) mmol/L,t =11.924,P =0.000].In early pregnancy,the risk of GDM increased by 7.984-fold (OR=8.984,95％CI:6.605-12.220) with every 1 mmol/L increase of the FPG level.(2) The diagnostic rate of GDM during mid-and last-trimester in group A (15.2％,370/2431) was lower than that of group B (43.9％,145/330),x2 =123.976,P =0.000.(3) Receiver operating characteristic curve analysis of FPG in early pregnancy and diagnosis of GDM:The largest area under the curve was 0.718 (95％ CI:0.690-0.747).The sensitivity and specificity were 0.600 and 0.612,or 0.735 and 0.726 respectively,when 4.795 mmol/L or 4.785 mmol/L were set as the cut-off value.(4) Among the 1208 cases delivered,GDM was diagnosed in 227 cases.The cesarean section rate (54.2％,123/227) of GDM women was higher than that (39.2％,385/981) of non-GDM women (x2 =16.884,P=0.000).There were no differences in the incidences of macrosomia,neonatal hyperbilirubinemia,low birthweight infant,premature delivery,fetal growth restriction and preeclampsia between GDM and non-GDM group (all P＞0.05).The incidence of premature birth in GDM women with FPG＜ 5.1 mmol/L was lower (5.8％,10/173) than that (14.8％,8/54) of women with FPG≥5.1 mmol/L (x2=4.601,P＜0.05).The incidence of cesarean section,insulin administration,macrosomia and preeclampsia increased from low FPG group to high FPG group,however there was no statistical significances.Conclusions Diagnosing GDM with FPG≥5.1 mmol/L in early pregnancy is not recommended as over diagnosis might happen.But this cut-off value might indicate that the patient are at risk of GDM,and this population should not be ignored.

AIM: To construct recombinant lentiviral vector with short hairpin RNA (shRNA) of CREB gene, and to investigate the effect of CREB gene silencing on mitochondrial morphology and cell apoptosis in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced cortical neurons.METHODS: Three lentiviral vectors pLentiLox3.7 (PLL) inserted shRNA fragments targeting CREB gene were co-transfected with the packaging plasmids psPAX2 and pMD2.G to the 293T cells, and the virus particles, which was infected with the primary cortical neurons, was encapsulated.The protein expression of CREB was detected by Western blot.The mitochondrial morphology, cell apoptosis and the expression of Bcl-2 and Bax were evaluated by the methods of MitoTracker red, TUNEL and Western blot in OGD/R induced cortical neurons after CREB gene silencing.RESULTS: The pLL-CREB-shRNA1 was the most effective shRNA, which inhibited 80% CREB gene expression in the cortical neurons.The mitochondrial was appeared dot and fragment morphology in OGD/R induced cortical neurons with transfected pLL-CREB-shRNA1 plasmid.In addition, the expression of Bcl-2 was decreased, the expression of Bax, and the apoptosis of the neurons were increased by tranfected with pLL-CREB-shRNA1.CONCLUSION: CREB shRNA recombinant lentiviral vector specifically inhibits the expression of CREB gene.CREB gene silencing promotes the cell apoptosis and mitochondrial morphological changes in the cortical neurons induced by OGD/R.

Objective To investigate the association between polymorphism of HLA-DRB1 allele and systemic sclerosis (SSc) and scleroderma-associated renal damage in Han Chinese of Henan Province.Methods Eighty-one SSc patients and 90 normal controls were recruited in this study,among them 59 patients had renal damage (SRD).The genotyping was carried out by nest PCR-SBT and gene clone.Comparisons between groups were performed with x2 test or exact probabilities.Multivariable Logistic regression was used to evaluate the association between prevalence of SSc or SRD and the possible relevant alleles.Results Thirty-three HLA-DRB1 alleles were discovered from the specimens,including 27 in SSc specimens,and 22 in SRD.Among them,the allele frequencies of HLA-DRB1 * 040501 (8.64％), * 110101 (11.11％), * 150201(8.02％) were higher in SSc patients than those of the controls (1.67％,4.44％,2.22％ respectively).After adjusted for other factors,HLA-DRBl * 040501 (P=0.010,OR =5.839,95％CI:1.518-22.460)、* 110101(P=0.019,OR=3.060,95％CI:1.204-7.772)、* 150201(P=0.010,OR=4.780,95％CI:1.444-15.821 )were identified as independent risk factors for SSc.And the allele frequencies of HLA-DRB1*040501 (9.32％),* 150201 (7.63％) were higher in SRD patients than those of the controls (1.67％,2.22％ respectively).After adjusted for other factors,HLA-DRB1 * 040501 (P=0.008,OR=6.363,95％CI:1.614-25.087) and * 150201 (P=0.030,OR =4.043,95 ％CI:1.147-14.243 ) were identified as independent risk factors for SRD.Conclusion Our data suggest that HLA-DRB1 * 040501,* 110101,* 150201 may be susceptible genes for SSc and the HLA-DRB1 * 040501,* 150201 may be susceptible genes for SRD in Han Chinese of Henan Province.

Objective To discuss the serum endoglin expression in severe pre-eclampsia and eclampsia women and their relationships. Methods Forty-two severe pre-eclamptic patients and 4 eclamptic patients in Peking University First Hospital from Dec. 2005 to Dec. 2007 were enrolled in the study group, with the mean gestational week of 35 ± 4, the mean age of 29.3 ± 5.7 and the mean BMI (30.1 ± 4.1 ) kg/ m2. This group included 25 cases of early onset pre-eclampsia, 21 cases of late onset pre-eclampsia, 8 cases of fetal growth restriction and 5 cases of HELLP syndrome. The control group included 29 cases of normal pregnant women during the same period, with the mean gestational week of 33±4, the mean age of 30.7± 3.4 and the mean BMI(27.2±2. 2) kg/m2. Peripheral serum endoglin was determined by ELISA in these two groups. Results (1)There is positive correlation between serum soluble endoglin level and the gestational weeks during 27 to 37 gestational weeks in the control group (r=0.79, P<0.05), but there is no distinct relationship in the study group (r=0.31, P>0.05). (2) Serum endoglin level of severe pre-eclampsia group was higher than the normal group [(14.2±5.6)μg/L vs. ( 10.9 ± 4.2 ) μg/L, P<0.05]. (3) Serum endoglin level of early onset group did not differ from late onset group [(14.3±5.7)μg/L vs. (13.6±5.0)μg/L, P >0.05]. (4) No difference of serum endoglin between HELLP group and non-HELLP group was found [(10.1±2.9) μg/L vs. ( 14.4±5.4) μg/L, P>0.05 ]. (5) Serum endoglin level of FGR sub group was higher than non-FGR sub group [(17.3±6.1) μg/L vs. (13.0±4.8) μg/L, P < 0.05] in the stady group. Conclusion The elevated peripheral serum endoglin level may contributes to the pathogenesis of severe pre-eclampsia and FGR, but not the week of the onset of the disease.

Objective To investigate the values and characteristics of 75 g oral glucose tolerance test (OGTT) in pregnant women.Methods A total of 6 103 singleton pregnant women aged (30.4±3.8) years (18-49 years) who delivered in Peking University First Hospital between May 1,2011 and December 31,2012 underwent the 75 g OGTT at gestational age of 24-28 weeks.They were divided into five groups based on maternal age:＜25 years (n=222,3.6％),25-years (n=2 485,40.7％),30-years (n=2 573,42.2％),35-years (n=683,11.2％),and ≥ 40 years (n=140,2.3％).The normal values of the fasting,1 h and 2 h blood glucose were lower than 5.1,10.0 and 8.5 mmol/L.Gestational diabetes mellitus (GDM) was diagnosed when blood glucose of any point was higher than or equal to normal value.Comparison between groups was tested by analysis of variance and LSD test.Logistic regression was used to calculate the risk for GDM in different age groups.Results (1) The fasting,1 h and 2 h blood glucose levels were in Gaussian distribution.The （-x）+2s were 5.51,11.12 and 9.49 mmol/L.The 97.5 percentile were 5.63,11.32 and 9.95 mmol/L.Fasting plasma glucose of ＜ 25,25-,30-,35-,and ≥ 40 years were (4.53±0.40),(4.60±0.40),(4.67±0.43),(4.74±0.46) and (4.82±0.49) mmol/L.The 1 h blood glucose were (6.98± 1.70),(7.55± 1.60),(7.92± 1.63),(8.30± 1.71) and (8.76± 1.86) mmol/L.The 2 h blood glucose were (6.11±1.33),(6.53±1.27),(6.89±1.33),(7.23±1.50) and (7.57±1.60) mmol/L.Therewas statistical difference in the blood glucose levels at a same time-point test among different age groups (F=29.61,60.17 and 72.29,all P＜0.01).(3) The total prevalence rate of GDM was 21.1％ (1 290/6 103) ; and the prevalence rates were 9.9％ (22/222),16.7％ (414/2 485),22.7％ (583/2 573),32.1％ (219/683) and 37.1％ (52/140) among the five age groups,respectively,with significant differences (x2=120.68,P=0.00).Compared with the group aged ＜25 years,the OR (95％CI) of the prevalence among 25-,30-,35-,and ≥40 years group were 1.82 (1.16-2.86),2.66 (1.70-4.18),4.29 (2.69-6.86) and 5.37 (3.08-9.39),respectively.Conclusions Advanced age is a risk factor for GDM.The risk of GDM increases significantly after 35 years old and pregnancy in women aged ＜ 35 years can reduce the risk of GDM.

Objective To understand the growth pattern of infants of mothers with maternal glucose metabolism during pregnancy.Methods Totally,7600 infants,born from singleton pregnant women from January 1st,2007 to December 31st,2009 in Peking University First Hospital and were followed up at 6-12 weeks after birth,were included.Altogether,645 mothers were complicated with hyperglycemia and 6955 with normal glucose metabolism during pregnancy.All infants were divided into four groups based on maternal glucose metabolism and their birth weight:Group N1 (n =6432) was consisted of non-macrosomia infants with normal maternal glucose metabolism; Group N2 (n =523) included macrosomia infants with normal maternal glucose metabolism; Group A1 (n =588) were non-macrosomia infants with abnormal maternal glucose metabolism; Group A2 (n =57) were macrosomia infants with abnormal maternal glucose metabolism.Birth weight,body weight at the day of follow-up and average daily weight gain were compared among these four groups.T-test,single variance analysis and LSD was applied in statistics,and the time at follow-up was used as co variance to find out the early growth pattern of infants.Results The birth weight of infants in normal and abnormal glucose metabolism group showed no statistical difference [(3367.0±420.3) g vs (3368.2±475.1) g,t=-0.061,P＞0.05],but body weight at the day of follow-up and the daily weight gain in the former group were lower than in the latter [body weight at follow-up:(5459.3±625.2) g vs (5393.9±647.2) g;daily weight gain:(44.0±9.5) g vs (42.9±9.5) g,t=2.464 and 2.874,all P＜0.05].The birth weight of infants in Group N1,A1,N2 and A2 was (3300.6±359.2) g,(3282.1±397.0) g,(4183.8±203.8) g and (4256.8±248.8) g,respectively;the body weight at the day of follow-up was (5400.5±590.7) g,(5325.8±618.8) g,(6182.7±584.7) g and (6096.5±502.4) g;daily weight gain was (44.1±9.4) g,(43.2±9.4) g,(42.4±10.9) g and (39.6±10.0) g,respectively (F=1140.471,313.376 and 10.830,all P＜0.001).While using co-variance to compare among the four groups,statistically more daily weight gain was shown in Group N1 than in A1,A2 and N2,in Group N2 than in Group A2,in Group A1 than in A2 (all P＜ 0.05).Conclusions The growth speed may slow down in early infantile period for offsprings of mother with hyperglycemia during pregnancy.

Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi's extracts. Methods Twelve Sprague-Dawley rats'right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium,NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi;the normal group contained the extracts of the normal hippocampi;the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs into neurons.

Objective To identify one runny mucoid-like Gram-negative bacteria with pink pigment isolated from clinical pus sample. Methods The pus sample was aseptically extracted from a deep lesions of one patient, then stored in Amies medium at room temperature for transportation. One sheep blood plate and one chocolate plate were used to detect the possible pathogens from the specimens. After inoculation, the plates were placed in a humidified incubator with 5% CO2 at 35 ℃. To identify the obtained isolates, we used the commercial Vitek2 and API systems, combining some traditional morphological examination and classical biochemical and physiological characteristics. For pure cultures, the cellular fatty acids were extracted, methylated, and determined by gas chromatography method. The 16S rRNA gene was amplified and sequenced by a commercial broad-spectrum PCR primers. The phylogenetic tree based on 16S rRNA gene was constructed by Mega 4.1 software using the neighbour-joining methods with 1 000 bootstrap replications. Results One runny mucoid-like Gram-negative bacterium, named K8756, was isolated both on sheep blood and chocolate plates after 72 h incubation. The API 20NE profile was 1245045 after a 3-day culture, which would be identified as Ochrobactrum anthropi with a good confidence of 98% probability. It was identified as Ralstonia pickettii and Bordetella bronchiseptica by VITEK 2 GN kits. However, further comparative 16S rRNA gene sequences showed that strain K8756 was closely related to the valid published Roseomonas mucosa MDA 5527 with 100% identity. Colonial morphologic features, phenotypic characteristics and major cellular fatty acid composition were also with high similarity to Roseomonas mucosa. Conclusions Strain K8756( = GIMCC 1.0030 ) is identified as Roseomonas mucosa by the polyphasic phenotypic and genotypic characteristics. The comparative analysis based on 16S rRNA gene sequences is a useful method for identifying the problematic and newly named bacteria.

Objective To detect the serum level of mannose binding lectin (MBL) in patients with renal injury induced by rheumatoid arthritis (RA), and to investigate the role of MBL in the pathogenesis of renal injury in RA. Methods ELISA was used to measure the serum MBL level of 19 RA patients with renal injury, 49 RA patients without renal injury and 40 healthy individuals. The clinical features and laboratory markers were compared and analyzed by chi-square test, two independent samples t-test and Spearman's correlation analysis. Results Compared with RA patients without renal injury, the number of tender and swollen joints [(15±9) vs (9±11)], duration of morning stiffness [(2.9±1.3) vs (2.3±1.6) h] and extraarticular manifestations (42.1％ vs 16.3％) in RA patients with renal injury were significantly higher (P＜0.05or P＜0.01). There was no significant difference between the two groups in RA disease duration and jointdeformity(P＞0.05). In patients with renal injury, the level of platelet count [(376±155)×109/L vs (304±121)×109/L], CIC[(4.3±3.0) vs (2.9±3.3) g/L], ESR[(79±46) vs (53±31) mm/1 h], RF[(77±42) vs (52±49)U/ml], CRP[(32±28)vs (23±18)mg/L], IgG[(11.7±2.6)vs (8.4±2.4)g/L], C3[(1.18±0.53)vs (0.94±0.21) g/L] were higher than those in RA patients without renal injury (P＜0.01 or P＜0.05); the level of Alb [(26±13) vs (30±9) g/L] was lower than that in RA patients without renal injury (P＜0.05). The level of serum MBL in the two groups of RA patients was significantly lower than that in the healthy group [(3.1±0.5)mg/L](P＜0.01), and the level of serum MBL in RA patients with renal injury [(1.7±1.2) mg/L] was higher than that in RA patients without renal injury [(1.4±1.3) mg/L](P＜0.05). The level of serum MBL in RA patients with renal injury showed a positive correlation with IgG, C3, CRP and 24 h urine protein level (r=0.6, 0.6, 0.47, 0.57; P＜0.05). Conclusion Renal injury in RA patients is immune complex dependent. The serum level of MBL is higher in patients with renal injury, therefore, high-concentration MBL may be one of a potential causes of renal injury in RA patients.