Objective To investigate the expression and significance of Rho-associated protein kinaseⅡ(Rock Ⅱ)in preeclamptic placenta and umbilical artery.Methods Semiquanfitative RT-PCR and Western blot were used to investigate the expression of RockⅡmRNA and RockⅡprotein in placenta and umbilical artery from 35 women with moderate preeclampsia(MPE group)、38 women with severepreeclampsia(SPE group)and 45 normal third trimester pregnant women(control group),the S/D value of umbilical artery was examined by ultrasound.Results (1)The expression of Rock Ⅱ mRNA of Dlacenta in MPE group(0.82±0.14)and SPE group(0.93±0.13)were signifieantly higher than that in control group (0.70 ±0.12,P＜0.01).The expression of Rock Ⅱ protein of placenta in MPE group(0.79±0.15)and SPE group(0.92±0.12)were significantly higher compared with control group(0.68±0.11,P<0.01).The expression of Rock Ⅱ mRNA and protein of placenta in SPE group were higher compared with MPE group(P＜0.01).(2)The expression of Rock Ⅱ mRNA of umbilical artery in MPE group(0.69±0.13)and SPE group(0.55±0.12)were significantly lower than that in control gmup(0.76±0.10,P＜0.01).The expression of RockⅡ protein of umbilical artery in MPE group(0.68±0.10)and SPE group(0.51±0.12)were lower compared with control group(0.75±0.13,P＜0.01).The expression of RockⅡ mRNA and protein of umbilical artery in SPE group were significantly lower compared with MPE group(P＜0.01).(3)There were no correlations between the expression of RockⅡ mRNA and protein in placenta and umbilical artery and the S/D value and birth weight(P＞0.05).Conclusion The upregulated expression of Hock Ⅱ in placentas and downregulated expression in umbilical artery may be a compensation in preeclampsia.

Objective To investigate the role of oxidized low-density lipoprotein (oxLDL) and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-I) in pre-ecalmpsia. Methods From June 2007 to January 2008,73 women with pre-eelampsia who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited, including 35 women with mild pre-eclampsia (MPE group) and 38 women with severe pre-eclampsia(SPE group). And 45 healthy pregnant women were taken as control group. Enzyme-linked immunosorbent assay (ELISA) was used to measure the plasma concentrations of oxLDL in these women. Semiquantitative RT-PCR and Western blot were used to investigate the expression of LOX-1 mRNA and protein in placenta. The expression of caspase-3 mRNA in placenta was determined using Semiquantitative RT-PCR. Results (1) The plasma concentrations of oxLDL in MPE group ( 0.42 ± 0.11 ) mg/L, SPE group ( 0.68 ± 0.12 ) mg/L, were significantly higher compared with control group (0.35 ± 0.14 )mg/L( P < 0.01 ) and the concentrations of oxLDL in MPE group were significantly higher compared with SPE group (P<0.01 ). (2) The expression of LOX-1 mRNA in placenta of MPE group(0.70 ±0.10) and SPE group(0.84 ±0.08) were significantly higher than that in control group(0.58 ± 0.11 ) ( P<0.01 ) and the expression of LOX-1 mRNA in MPE group was significantly higher than that of SPE group (P<0.01 ). (3) The expression of LOX-1 protein in placenta of MPE group (0.79±0.15 )and SPE group(0.90±0.12) were significantly higher compared with control group(0.68 ±0.11)( P<0.01 ), while the expression of LOX-1 protein of MPE group was significantly higher compared with SPE group (P <0.01 ). (4) The expression of caspase-3 mRNA in placenta of MPE group(3.82± 0.18) and SPE group(5.39±0.14) were significantly higher than that in control group(2.19±0.20) (P <0.01 ), and the expression of caspase-3 mRNA in MPE group was significantly higher than that of SPE group (P<0.01 ). (5) In pre-eclampsia groups, the levels of LOX-1 mRNA in placenta were positively correlated with the plasma concentrations of ox LDL ( r=0.93, P<0.05 ), and caspase-3 mRNA expression were positively correlated with the expression of LOX-1 mRNA in placenta( r=0.84, P<0.05). Conclusion Increased levels of oxLDL in plasma may induce excess expression of LOX-1 in placenta, which may be involved in the pathophysiological processes of pre-eclampsia.

Objective To investigate the expression and significance of serum soluble fms-like tyrosine kinase 1 ( sFlt-1 ) in preeclampsia placenta. Methods Immunohistochemistry and semi-quantitative RT-PCR were used to investigate the expression of sFlt-1 and sFlt-1 mRNA in placenta of 30 women with preeclampsia (including mild and moderate preeclampsia 11 cases, severe preeclampsia 19 cases) and 45 normal pregnant women (including first trimester pregnancy 18 cases, 2nd trimester pregnancy 12 cases, and 3rd trimester pregnancy 15 cases). The levels of vascular endothelial growth factor(VEGF) and sFlt-1 in serum were detected by enzyme linked immunosorbent assay. Results (1) The expression of sFlt-1 mRNA was significantly higher in preeclampsia group (0. 90?0. 11) compared with the third trimester group (0. 80?0. 06; P

Objective To investigate the correlation of the expressions of angiopoietin-2 (Ang-2) and angiopoietin-2 receptor(Tie-2)in serum and placenta with preeclampsia. Methods From May 2009 to April 2010, 62 women with preeclampsia who delivered in Affiliated Hospital of Qingdao University Medical College were recruited in the study, including 30 women with moderate preeclampsia (MPE group) and 32 women with severe preeclampsia (SPE group). Another 30 healthy pregnant women were taken as control group. ELISA was used to measure the serum Ang-2 in these women. Semiquantitative reverse transcription (RT)-PCR was used to investigate the expressions of Ang-2 mRNA and Tie-2 mRNA in placenta. Western blot was used to determine the expression of Ang-2 protein in placenta. Results (1) The serum concentrations of Ang-2 in MPE group and SPE group were (5.4 ± 1.8) μg/L and (5. 1 ± 1.7) μg/L,respectively. Both were significantly lower than that in control group (16. 2 ± 4. 5) μg/L (P<0. 01).There was no significant difference between MPE group and SPE group (P > 0. 05). (2) The expressions of Ang-2 mRNA in placenta of MPE group (2. 1 ± 0. 7) and SPE group (2. 0 ± 0. 6) were both significantly lower than that of control group (5.8 ± 0. 8; P<0. 01). But there was no significant difference in Ang-2 mRNA expression between MPE group and SPE group (P>0. 05). (3) No significant difference was found in the expressions of Tie-2 mRNA in placenta among MPE group (1. 33 ±0. 04), SPE group (1.35 ±0. 05) and control group (1.34 ± 0. 04; P > 0. 05). (4) The expressions of Ang-2 protein in placenta of MPE group (2.0 ± 0. 8) and SPE group (2. 0 ± 0. 8) were both significantly lower than that of control group (5.7 ±0. 9; P <0. 0l), while no significant difference was found between MPE group and SPE group (P >0. 05) . (5) In MPE group and SPE group, the serum concentrations of Ang-2 were positively correlated with the levels of Ang-2 mRNA and Ang-2 protein in placenta(r =0. 651, 0. 627; P <0. 01). Conclusions Decreased expressions of Ang-2 mRNA and Ang-2 protein in placenta reduced serum concentration of Ang-2. Low expression of Ang-2 may be involved in the pathophysiological process of preeclampsia by affecting the formation of placenta in early pregnancy.

Objective To assess the diagnostic predictive value of Wells score and modified Geneva score for acute pulmonary embolism by prospective case series and to explore a more suitable scoring system for Chinese population.Methods All the patients suspected of pulmonary embolism (PE) and received CT pulmonary angiography (CTPA) were enrolled consecutively in Fuxing Hospital,Capital Medical University,China,from June 2009 to August 2011.Before CTPA test or on condition that test results were unknown,clinical scoring was assessed prospectively by the Wells score and the modified Geneva score.The probability of PE in each patient was assessed and the patients were divided into low,moderate and high probability groups according to the clinical scores.The result of CTPA was used as the diagnostic gold standard for PE.Diagnostic accuracy in each group was analyzed.The predictive accuracy of both scores was compared by AUCROC curve.Results A total of 139 patients met our enrollment criteria and 117 eligible patients entered our study at last.PE was diagnosed in 47 patients by CTPA with an overall prevalence of 40.2％.Prevalence of PE in the low,moderate and high pretest probability groups assessed by the Wells score and by the simplified modified Geneva score were 7.1％ (3/42),42.9％ (21/49),88.5％ (23/26)and 10.0％ (3/30),48.1％ (37/77),7/10,respectively.AUCROC curves for the Wells score and the simplified modified Geneva score were 0.872 ( 95％ CI 0.810-0.933 ) and 0.734 ( 95％ CI 0.643-0.825 )respectively,with a significant difference ( P =0.005 ).Conclusion The Wells score is more accurate for clinical predicting acute PE than the modified Geneva score.

BACKGROUND: Key point for subculture of human embryonic stem cells (hESCs) is to inhibit spontaneous differentiation and make sure totipotency of cells. Although mouse embryonic fibroblasts (MEF) or human foreskin fibroblasts (HFF) used as the feeder layer can maintain undifferentiated state of embryonic stem cells, cell clone is still imperfect and parallel arranged. OBJECTIVE: To establish mixed feeder layer of mouse embryonic fibroblasts plus human foreskin fibroblasts and to observe the hESCs growth.DESIGN: Multi-sample observation and comparison.SETTING: Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College. MATERIALS: This study was performed at the Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College from April 2006 to July 2007. Foreskin was derived from the children who underwent circumcision and came from Urinary Surgery, the Affiliated Hospital of Haihan Medical College. The children's family members provided the informed consent, and the experiment received confirmed consent from the local ethic committee. hESCs line HN-1 was separated from human blastula. Eleven 12.5-14.5-day-old fetal mice of clean grade were selected in this study. The experimental animals were disposed according to ethical criteria. METHODS: Heads, four extremities, and viscera were removed from fetal mice under anesthesia. Subsequently, cell suspension was prepared using routine trypsinase digestion and inoculated. When cells were cultured in confluent monolayer, some primary cells were frozen, processed with mitocin-C for 2.0-3.0 hours, and inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., MEF feeder layer. The HFF separation and culture and the preparation of HFF feeder layer were the same as above-mentioned processing. In addition, the two fibroblasts were respectively counted and mixed together according to the ratios of 1∶0, 3∶1, 1∶1, 1∶3, and 0:1. And then, the mixture was inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., mixed feeder layer. The growth of subcultured hESCs in vitro was observed in three different feeder layers, and hESCs in the mixed feeder layer underwent alkaline phosphatase test, OCT-4 expression immunohistochemical measurement, and OCT-4 and telomerase mRNA expression RT-PCR detection. Finally, differentiation in vitro of hESCs was observed after removing the feeder layer.MAIN OUTCOME MEASURES: ① Growth of hESCs in the three different feeder layers; ② Growth of hESCs in the mixed feeder layer based on different mixed ratios; ③ undifferentiated state of hESCs in the mixed feeder layer; ④ differentiation in vitro.RESULTS: ① hESCs clone in the MEF and HFF feeder layers was thin and flat and imperfect, but hESCs clone in the mixed feeder layer was perfect and thick and solid. Apparently, the clone form in the mixed feeder layer was superior to MEF and HFF feeder layers. ② When MEF and HFF was mixed together according to the ratio of 1∶1, hESCs grew in apparent accumulation; clone border was clear; eminentia was apparent and perfect. However, there were no changes according to the ratio of 1∶3. The ratio of 1∶1 was superior to the ratios of 1∶0, 3∶1, and 0∶1. ③ Alkaline phosphatase staining and OCT-4 antigen expression were strongly positive. Specific straps of OCT-4 and telomerase mRNA expression were observed at 200-300 bp and 300-400 bp, respectively. ④ Embryoid bodies were formed. hESCs could differentiate into multi-morphological cells after attachment.CONCLUSION: ① The mixed feeder layer may well support in vitro subculture of hESCs to acquire excellent clone form compared to MEF or HFF feeder layer. ② The mixture of MEF and HFF has excellent effect according to the ratio of 1∶1.

Objective To investigate the value of combined 3D-constructive interference in steady state(3D-CISS) with phase-contrast MR(PC-MR) in diagnosis of hydrocephalus children.Methods We collected retrospectively 28 children with hydrocephalus who caused by obstructive membranes.They all underwent MR imaging examination with a 1.5 T consisting of 3D-CISS, conventional sequences and PC-MR.With the result of surgery and CT cisternography as gold standard,we analysised different sequence in the diagnosis of membranous obstruction.The results of obstructive were analysed by x2 test between 3D-CISS combined with PC-MR and conventional sequences.Results The study included 28 children, of whom 16 showed obstructive membranes in CSF pathways of ventricle and 12 displayed obstructive membranes in CSF pathways of cistern.Conventional sequences diagnosed obstructive membranes of 15 children, and obstructive membranes of 11 children were consistent with gold standard, and 4 cases were false positive.The 3D-CISS diagnosed obstructive membranes of 40, and 12 cases were false positive.There were significant difference between 3D-CISS combined with PC-MR and conventional sequence about obstructive membranes diagnosis of CSF pathways(x2=24.410, P=0.001).Conclusions Making use of jointing 3D-CISS sequence and PC-MR sequence successively, and we can diagnose the position of obstructive membrane for hydrocephalus, which can be helpful to select a perfect surgical approach.

Objective To observe the clinical effect of combination of Tongxiening granule and Mesalazine on treating mild and moderate ulcerative colitis(UC) .Methods Totally 380 patients with mild‐to‐moderate UC diagnosed through endoscopy were allocated to the control group (n=190) and observation group(n=190) .For the observation group ,patients were remedied with the combination of Tongxiening Granules and the Mesalazine by oral administration for eight weeks ,meanwhile the control group only received the Mesalazine for eight weeks .The total effective rate of the two groups were statistically analyzed ,and the levels of ser‐um MMP‐2 and MMP‐9 before and after treatment in the two groups were measured .The expression of S100A12 and RAGE were detected by immunohistochemistry SP method .Results The total effective rate of the observation group and the control group was 94 .74% and 89 .47% respectively ,and the difference was statistically significant(P<0 .01) .After treatment ,the expression levels of MMP‐2 and MMP‐9 in the two groups were decreased ,additionally the expression levels in the observation group was lower than those in the control group ,and the difference was statistically significant (P< 0 .01) .After treatment ,the expression levels of RAGE and S100A12 in the observation group were decreased ,and there was a significant difference when compared with the control group(P<0 .01) .Conclusion Combined application of Tongxiening Granules and Mesalazine in treating patients with mild‐to‐mod‐erate UC could better improve clinical symptoms and bring better therapeutic effect than single use of Mesalazine .

Objective To investigate the relationship between serum visfatin levels and glycometabolism in gestational diabetes mellitus(GDM). Methods 80 cases pregnant women were divided into two groups: GDM group and normal glucose tolerance group(control group). The levels of serum visfatin、fast insulin(FIN)、fast plasma glucose(FPG)、1h、2h、3h plasma glucose after glucose loading(1hPG、2hPG、3hPG )were measured. Insulin resistance index(HOMA-IR) was calculated. Results (1)The levels of visfatin、FPG、1hPG、2hPG、3hPG、HOMA-IR、FIN were significantly higher in GDM group than control group(P＜0.01). (2)There was a positive correlation between visfatin and HOMA-IR and 2hPG(P＜0.01). Multiple regression analysis showed that 2hPG was independent related factor. Conclusion Increased serum levels of visfatin represents a compensatory mechanism aimed at high glucose levels in women with GDM.[Key Words] Visfatin;Diabetes;Gestational;Insulin resistance

Osseointegration plays an important role in the functions and aesthetics of dental implant. This paper focus on the hormones closely related to the bone metabolism, including glucocorticoid, parathyroid hormone, calcitonin, melatonin, and oestrogen, and reviews the influence of hormones on the osseointegration.