Temporary Internal Iliac Artery Blockage versus Uterine Artery Embolization in Patients After Laparoscopic Pregnancy Tissue Removal Due to Cesarean Scar Pregnancy.

OBJECTIVE: This study aims to evaluate the clinical value of laparoscopic temporary internal iliac artery blockage (TIIAB) compared with uterine artery embolization (UAE) in type III cesarean scar pregnancy (CSP). METHODS: A total of 76 patients with type III CSP admitted to the Department of Gynecology the First Affiliated Hospital of Zhengzhou University between September 2017 and June 2019 were selected for this retrospective study. Thirty-six of them in the study group received TIIAB, and the rest in control group received UAE. Laparoscopic pregnancy tissue was removed from all patients, and the uterine defects were repaired. The absence of remnants was then confirmed using ultrasonography. Follow-ups were performed in the two groups for six months, and the factors of intraoperative blood loss, operation and menelipsis time, 24-h human chorionic gonadotropin decline rate, postoperative complications, hospitalization days, hospitalization costs, peri-operative hormone levels, and ovarian function indicators were compared between the two groups and within each group. RESULTS: There were statistically significant differences in the hospitalization cost, menelipsis time, and postoperative complication incidence between the two groups (p < 0.05). There were statistically significant differences between ovarian function at one month and three months after surgery (p < 0.05) as well as among the follicle-stimulating hormone, luteinizing hormone, and estradiol levels at one, three, and six months after surgery in the control group (p < 0.05). CONCLUSION: Compared with uterine artery embolization, laparoscopic TIIAB has the advantages of a low hospitalization cost, lower postoperative complication rate, and shorter menelipsis time. Moreover, it avoids ovarian function damage. It is a safe method worthy of clinical popularization.

Intermittent hypoxia enhances the tumor programmed death ligand 1 expression in a mouse model of sleep apnea.

BACKGROUND: As a hallmark of obstructive sleep apnea (OSA), intermittent hypoxia (IH) promotes tumor progress. The high expression of programmed death 1 and programmed death ligand 1 (PD-L1) in tumor leads to immune evasion and subsequently aggravates tumor progress. This study aims to determine the tumor PD-L1 expression under the IH condition. METHODS: A total of 24 C57BL/6J mice were randomly assigned to the normoxia (control, CTL) group and the IH group. Mice in the IH group were subjected to the IH condition for 5 weeks. Lung cancer cells were injected into the flank of each mouse after 1 week of IH exposure. Tumor PD-L1 expression was detected by immunohistochemistry (IHC). Correlation between tumor weight, tumor volume, and expression of PD-L1 was analyzed. RESULTS: Compared to the CTL group, mice in the IH group had a high PD-L1 expression. The IH can enhance the tumor PD-L1 expression. Tumor weight, volume, and HIF-1α levels were closely associated with the PD-L1 expression in the IH group, while dissimilar findings were observed in the CTL group. CONCLUSIONS: The IH enhances tumor PD-L1 expression in OSA mimicking mice. Additional studies are required to clarify the underlying mechanism.

Tapered Cuff versus Conventional Cuff for Ventilator-Associated Pneumonia in Ventilated Patients: A Meta-Analysis of Randomized Controlled Trials.

Background: Microaspiration of secretions around the tracheal cuff is a multifactorial process. Tracheal cuff shape might take a major part in its occurrence. The rationale for producing a taper-shaped cuff is established on the assumption that compared to a conventional cuff with a single fixed diameter, a continuum of minimum-to-maximum diameter sections might better fit the tracheal walls. Objectives: The primary objective of this meta-analysis was to compare ventilator-associated pneumonia (VAP) between tapered-cuff intubation and conventional-cuff intubation. The secondary objective was to compare intensive care unit (ICU) mortality between tapered-cuff intubation and conventional-cuff intubation. Methods: We searched the Cochrane Library, Embase, MEDLINE database through the PubMed search engine, and CINAHL from inception to April 2018. Randomized trials comparing VAP and ICU mortality between tapered-cuff intubation and conventional-cuff intubation in intubated adults were included. Two review authors assessed study quality and abstracted databasing on prespecified criteria independently. Results: We pooled summary estimates from 5 trials evaluating tapered-cuff involving 774 participants. Compared to VAP, no statistically significant difference was observed between the tapered-cuff and conventional-cuff groups (OR 0.82, CI 0.61-1.12, z = 1.24, and p=0.21). No statistically significant difference was observed between the tapered-cuff and conventional-cuff groups with ICU mortality (OR 0.77, CI 0.55-1.08, z = 1.49, and p=0.14). Conclusions: In this meta-analysis, the tapered-cuff tracheal tube may not be superior to the standard-cuff tracheal tube in reducing VAP and ICU mortality.

Comparing the effects and compliance between volume-assured and pressure support non-invasive ventilation in patients with chronic respiratory failure.

INTRODUCTION: Standard bi-level non-invasive ventilation with fixed-level pressure support (PS) delivery may not maintain ventilation during the changes in pulmonary mechanics that occur throughout day and night, so average volume-assured pressure support (AVAPS) modes that target a preset volume by adjustment of PS may be effective. OBJECTIVE: Our meta-analysis wants to compare AVAPS and pressure support non-invasive ventilation (PS-NIV) regarding arterial blood gases (ABGs), sleep efficiency and compliance. METHOD: Relevant publications indexed in PubMed, Cochrane Library, Embase, Web of Science, Wanfang Data, China National Knowledge Infrastructure and VIPI were identified. Appropriate articles identified from the reference lists of the above searches were also reviewed. We included randomized controlled trials involved the use of AVAPS and PS-NIV ventilation for chronic respiratory failure. Each included study weighted mean differences, and 95% confidence intervals (CI) were calculated for continuous outcomes. Statistical heterogeneity was assessed using the I2 value ≤ 50% were considered as no statistical heterogeneity and used fixed effects model. Otherwise, a random effects model was used. RESULTS: Eight trials were eligible. No significant difference was observed between AVAPS and PS-NIV groups to compare PaCO2 (OR -0.97, CI-2.54-0.61, P = 0.23) and PaO2 (OR -1.81, CI-4.29-0.67, P = 0.15) in ABGs. There was no significant difference between the two groups with sleep efficiency (OR -3.31, CI-7.58-0.95, P = 0.13) and visual analog scale (OR 0.32, CI-6.97-7.61, P = 0.93). CONCLUSIONS: The evidence shows there is no significant difference in clinical outcomes when comparing AVAPS and PS-NIV used for chronic respiratory failure patients.

Influence of subglottic secretion drainage on the microorganisms of ventilator associated pneumonia: A meta-analysis for subglottic secretion drainage.

BACKGROUND: The influence of the subglottic secretion drainage (SSD) on the microorganisms of ventilator associated pneumonia (VAP) is still unclear.A meta-analysis focusing on the influence of the SSD on the microorganisms of VAP. METHODS: A comprehensive search was conducted through the online studies of PubMed, Embase, Cochrane Library, Google scholar, CNKI (China National Knowledge Infrastructure), and VIPI (Database for Chinese Technical Periodicals) using specific search terms.Included studies were randomized controlled trials (RCTs) that compare the microorganisms of VAP between SSD and standard endotracheal tube care in mechanically ventilated adults. RESULTS: Nine RCTs were eligible. There was no significant difference in the rate of VAP caused by nonfermentative bacteria and enterobacteria between SSD group and control group (OR = 0.73, 95%CI, 0.53-1.01; P = .06). The episodes of VAP caused by Gram-positive cocci and Haemophilus influenzae organisms were lower in the SSD group (OR = 0.29, 95%CI, 0.18-0.48; P<0.00001). Less mean volume of SSD daily was observed in VAP group (OR = -16.97, 95%CI, -29.87-4.08; P = .010). CONCLUSION: We found SSD to be associated with significant decreases in VAP caused by Gram-positive cocci and H influenzae organisms but no significant differences in VAP caused by nonfermentative bacteria and enterobacteria. Less mean volume of SSD daily was observed in VAP group.

Serum autoantibodies against a panel of 15 tumor-associated antigens in the detection of ovarian cancer.

In this study, enzyme-linked immunosorbent assay has been used to examine the frequencies of serum autoantibodies against two candidate tumor-associated antigens intensively selected from the Human Protein Atlas database, in combination with 13 tumor-associated antigens available from our lab in sera from 44 OC patients and 50 normal healthy controls. Conventional evaluation (mean + 3SD as the cutoff value to determine a positive reactivity), receiver operating characteristic curve analyses, and classification tree analysis were further used to evaluate the diagnostic performance of autoantibodies against these tumor-associated antigens (anti-tumor-associated antigens) in ovarian cancer. For single anti-tumor-associated antigen, when the cutoff values were set as mean + 3SD of normal healthy controls, NPM1, MDM2, PLAT, p53, and c-Myc could achieve sensitivity higher than 20% at 98% specificity. Combinational utilization of autoantibodies against MDM2, PLAT, NPM1, 14-3-3 Zeta, p53, and RalA achieved the optimal diagnostic performance with 72.7% sensitivity at 96% specificity. Receiver operating characteristic curve analysis showed that the area under the receiver operating characteristic curves of autoantibodies against c-Myc, NPM1, MDM2, p16, p53, and 14-3-3 Zeta were greater than 0.80. This indicated that these tumor-associated antigens held high potential to serve as diagnostic biomarkers in ovarian cancer detection. Decision tree analysis indicated that anti-c-Myc held high potential in the detection of ovarian cancer. Further studies are warranted to validate the diagnostic performance of these anti-tumor-associated antigens with high area under the receiver operating characteristic curve, including autoantibodies against c-Myc, MDM2, PLAT, NPM1, 14-3-3 Zeta, p53, and RalA.

Proteomic-based identification of HSP70 as a tumor-associated antigen in ovarian cancer.

Ovarian cancer commonly presents without prominent symptoms and is consequently diagnosed at advanced stages with unfavorable prognosis. Novel serological biomarkers for the early detection and clinical management of ovarian cancer are imminently needed. Proteomic-based methods for biomarker discovery are promising strategies implemented in cancer research. The aim of the present study was to identify new tumor antigens from the ovarian cancer cell line SKOV3 and their associated autoantibodies in sera of patients with ovarian cancer employing proteomic-based approaches. Proteins from the ovarian cancer cell line SKOV3 were extracted by two­dimensional polyacrylamide gel electrophoresis (2-DE) followed by western blotting and antibody reaction with sera from patients with ovarian cancer and normal controls. Positive spots were excised from Coomassie blue­stained gels and identified by liquid chromatography­tandem mass spectrometry (LC-MS/MS). The 2-DE analysis results revealed a total of 14 protein spots on the gel, and 7 proteins were finally identified by LC-MS/MS. In the subsequent experiment, using immunoassay on ovarian cancer sera and tissue-array slides, the well-known protein HSP70 was selected in order to validate this proteomic-based approach. In conclusion, the proteomic method used in the present study is a powerful instrument for identifying novel serum markers that may exhibit clinical usefulness in cancer.

miR424-5p functions as an anti-oncogene in cervical cancer cell growth by targeting KDM5B via the Notch signaling pathway.

AIMS: Aberrant expression of miRNAs exert the critical roles in carcinogenesis, including cervical cancer. Recent study corroborated the down-regulation of miR424-5p in uterine cervix adenocarcinoma. This research aimed to investigate the function and underlying mechanisms of miR424-5p in cervical cancer cell growth. MAIN METHODS: Tissues samples were collected from patients with cervical cancer and healthy control. The expression levels of miR424-5p were determined by qRT-PCR. After transfection with miR424-5p mimics or inhibitor, cervical cancer cell proliferation and apoptosis were evaluated by WST-1 and flow cytometry assay, respectively. The underlying mechanism involved in aforementioned processes was also explored. KEY FINDINGS: Expression of miR424-5p was notably decreased in cervical cancer tissues and cells. Overexpression of miR424-5p restrained cell proliferation and promoted cell apoptosis, but with little function in miR424-5p inhibitor-treated groups. Furthermore, KDM5B was identified as a direct target of miR424-5p as the evidence that miR-424-5p inhibited KDM5B expression and luciferase activity of KDM5B 3'-UTR. Here, KDM5B elevation majorly reversed miR424-5p-triggered inhibition in cell proliferation and increase in cell apoptosis. Moreover, silencing KDM5B expression also restrained cell growth. Additionally, miR424-5p overexpression inhibited the expression of Notch1 and Notch2, which was obviously rescued after KDM5B up-regulation. Simultaneously, blocking KDM5B also attenuated the activation of Notch pathway. Importantly, treatment with Notch agonist Jagged1 antagonized miR424-5p-mediated suppression on cell growth. SIGNIFICANCE: This research suggests that miR424-5p may act as a novel anti-oncogene in cervical cancer by blocking cell growth through targeting KDM5B-Notch pathway. Accordingly, our study will support a promising therapeutic strategy against cervical carcinoma.

[Effects of plasma from patients with preeclampsia on the proliferation and apoptosis of human umbilical vein endothelial cells and its relationship with lysophosphatidic acid receptors].

OBJECTIVES: To investigate the effects of plasma from the patients with preeclampsia on proliferation and apoptosis of human umbilical vein endothelial cells (HUVEC), and to explore the relationship between cell damage and lysophosphatidic acid (LPA) receptors. METHODS: Sixty patients with preeclampsia were recruited from October 2011 to June 2012 in the First Affiliated Hospital of Zhengzhou University. Among them, thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group. The other thirty healthy pregnant women were recruited in the healthy pregnant women group. The levels of plasma LPA in the three groups were measured. The HUVEC were cultured in vitro with plasma from the three groups, and a blank control group was set up as well. Proliferation and apoptosis of HUVEC were measured by MTT assay and flow cytometry. Immunohistochemistry of biotin streptomyces protein peroxidase (SP) method was used to measure the protein expression level of Edg 2, 4, 7. RESULTS: (1) The plasma LPA levels in the healthy pregnant woman group, mild preeclampsia group and severe preeclampsia group were (3.38 ± 2.08) µmol/L, (6.12 ± 0.22) µmol/L, (9.10 ± 0.17) µmol/L, respectively. The plasma levels of LPA in patients with preeclampsia were significantly higher than that in the healthy pregnant women (P < 0.01). (2) The proliferation rate of HUVEC in the mild and severe preeclampsia groups [(65.2 ± 2.7)% and (51.9 ± 2.8)%] were significantly lower than that in the healthy pregnant women group and the control group [(84.3 ± 3.1)% and (100.0 ± 0.0)%, P < 0.01]. (3) The early apoptosis rate, middle-late apoptosis rate and total apoptosis rate of HUVEC in the mild and severe preeclampsia groups [total apoptosis rate were (30.4 ± 2.0)% and (43.4 ± 2.5)%] were significantly higher than those in the healthy pregnant women group and the control group [total apoptosis rate were (18.6 ± 1.6)% and (8.0 ± 1.5)%, P < 0.01]. (4) The expression positive rates of Edg 2, 4, 7 proteins in the four groups were as following: mild preeclampsia group 83%, 80% and 73%; severe preeclampsia group 97%, 93% and 90%; healthy pregnant women group 40%, 40% and 37%, and the control group 10%, 10% and 7% respectively. The positive rates of HUVEC in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group (P < 0.01). CONCLUSIONS: The plasma of patients with preeclampsia could inhibit proliferation and promote apoptosis of HUVEC, and induce the expression of Edg 2, 4, 7 proteins. It suggested that the increase of lysophosphatidic acid in plasma could be one of the reasons of endothelial cell damage in patients with preeclampsia.

[Influence of pertussis toxin on GPER-mediated activation of phosphatidylinositol 3-kinase/protein kinase B signaling induced by 17ß-estradiol in endometrial carcinoma cells].

OBJECTIVE: To investigate the influence of pertussis toxin (PTX) on G protein-coupled estrogen receptor (GPER)-mediated activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling activated by 17ß-estradiol (17ß-E2) in endometrial carcinoma cells. METHODS: Expressions of GPER protein were detected by immunohistochemical SP method in Ishikawa and HEC-1A cells. Changes of levels of GPER, ERα and ERß protein and the activation of Akt protein were observed by western blot in the two cells after they were treated by PTX for 30 minutes at different concentrations (0, 0.1, 0.5, 1.0 µg/ml), and then co-stimulated with with 1×10(-6) mol/L 17ß-E2 respectively at different time (Ishikawa 30 minutes, HEC-1A 15 minutes). RESULTS: (1) Immunohistochemical SP method showed that GPER was positive stained in cell cytoplasm of Ishikawa and HEC-1A cell. (2) After co-treated with PTX at different concentrations (0, 0.1, 0.5, 1.0 µg/ml) and 10(-6) mol/L 17ß-E2, in Ishikawa cell, the ratio of p-Akt/Akt was 0.74 ± 0.54, 0.34 ± 0.06, 0.18 ± 0.03, 0.07 ± 0.15, the gray values of GPER was 0.872 ± 0.490, 0.395 ± 0.054, 0.145 ± 0.014, 0.034 ± 0.008, and with increasing concentration of PTX, the ratio of p-Akt/Akt and the expression of GPER decreased gradually (P < 0.05), which was most obviously when the concentration was 1.0 µg/ml (F = 63.729, P = 0.0001; F = 160.284, P = 0.0001); ERα and ERß protein had no significant change among different groups (P > 0.05). In HEC-1A cell, the ratio of p-Akt/Akt was 0.73 ± 0.09, 0.26 ± 0.14, 0.11 ± 0.03, 0, the Gray values of GPER is 0.927 ± 0.134, 0.485 ± 0.022, 0.194 ± 0.004, 0, and with increasing concentration of PTX, the ratio of p-Akt/Akt and the expression of GPER decreased gradually (P < 0.05), which were also completely inhibited when the concentration was 1 µg/ml (F = 1039.321, P = 0.0001; F = 109.646, P = 0.0001), ERα protein had no significant differences (P > 0.05) among different groups. ERß was negatively expressed. CONCLUSION: The results proposed that the activation of PI3K/Akt signaling in Ishikawa and HEC-1A cells could be inhibited after blocking the role of GPER by PTX.

[Characteristics of N forms and N-transforming bacteria in paddy soil under different tillage patterns].

Paddy soil samples were collected in layers (0-5, 5-12, and 12-20 cm) during rice growth period to investigate the characteristics of the N forms and N-transforming bacteria in the soil profile under different tillage patterns (no-tillage with straw returning, NTS; conventional tillage with straw returning, CTS; no-tillage, NT; and conventional tillage, CT). In the whole rice growth period, ammonifying bacteria in 0-5 cm soil layer had the highest number under NTS, and nitrosobacteria in 0-5 cm and 5-12 cm soil layers were more abundant but in 12-20 cm soil layer were lesser under CT than under NT. Nitrosobacteria and denitrobacteria in 0-20 cm soil layer were lesser under NTS than under CTS. At elongating and ripening stages, anaerobic N-fixing bacteria in 0-5 cm soil layer were more abundant under NT than under CT. In the whole rice growth period, the alkali-hydrolyzable N and total N contents in 0-5 cm soil layer were significantly higher but in 5-12 cm and 12-20 cm soil layers were lower under NT than under CT, and the NH4(+)-N and NO3(-)-N contents in 0-20 cm soil layer were higher under NTS but in 12-20 cm soil layer had no significant differences between NT and CT. Correlation analysis and multiple polynomial regression analysis further revealed that there were significant relationships between soil NH4(+)-N and soil ammonifying bacteria, nitrosobacteria and denitrobacteria, and between soil alkali-hydrolyzable N and soil anaerobic N-fixing bacteria. Among the test tillage patterns, NTS could be the more desirable one for the N supply and fertility maintenance of paddy soil.

[Expression and significance of aquaporin 1 in placenta, placental membranes and peritoneum of patients with hypertensive disorder complicating pregnancy].

OBJECTIVE: To explore the role of aquaporin 1 (AQP1) in the initiation and development of hypertensive disorder complicating pregnancy (HDCP), and to analyze the relationship between AQP1 expression and ascites formation of patients with eclampsia. METHODS: Sixty inpatients with HDCP were recruited in the study, including 20 patients with gestational hypertension, 20 with mild preeclampsia, and 20 with severe preeclampsia. And 20 healthy pregnant women were taken as control. Immunohistochemistry was used to analyze AQP1 expressions in placenta, embryolemma and peritoneum, and B-mode ultrasonography was used to detect the ascites level of the patients. RESULTS: (1) AQP1 expression was detected in placenta, embryolemma and peritoneum. AQP1 was mainly located in endotheliocytes of blood vessels and blood capillaries in placenta, endothelial cells of amniotic membrane in embryolemma, and endotheliocytes of blood capillary and small veins in peritoneum. (2) The ascites incidence of HDCP patients (63%, 38/60) was higher than that of controls (10%, 2/20; P < 0.01). (3) The positive expressive rate of AQP1 in placenta of patients with HDCP (85%) was higher than that of controls (70%, P < 0.01). Furthermore, the AQP1 positive expressive rate from severe preeclampsia (90%) was obviously higher than that from gestational hypertension patients (80%, P < 0.05). (4) The AQP1 positive expressive rate in embryolemma from HDCP patients (87%) was lower than that of controls (95%, P < 0.05). The expressive rate from severe preeclampsia patients (80%) was obviously lower than that from gestational hypertension patients (95%, P < 0.05) and that of controls. (5) The AQP1 expressive rate in peritoneum from HDPC patients (82%) was higher than that of controls (70%, P < 0.01). The expressive rate of AQP1 from severe preeclampsia patients (90%) was obviously higher than that from gestational hypertension patients (75%, P < 0.05) and that of controls. CONCLUSIONS: The expression level of AQP1 of patients with preeclampsia increases in placenta and peritoneum and decreases in embryolemma, and holds correlation with the degree of HDCP. All these suggest that the changes in AQP1 expression may play an important role in the initiation and development of HDCP and may be one of the mechanisms for ascites formation.

Increased staining for phosphorylated AKT and nuclear factor-kappaB p65 and their relationship with prognosis in epithelial ovarian cancer.

AKT plays an important role in malignant behavior of tumors. The purpose of the present study was to determine the expression of phosphorylated AKT (P-AKT) and nuclear factor-kappaB (NF-kappaB) p65 and their association with clinicopathological parameters and prognosis in epithelial ovarian tumor. On immunohistochemistry 115 samples of ovarian tissue that included 68 specimens of epithelial ovarian cancer, 12 of borderline tumor, 24 of epithelial benign tumor and 11 of normal ovary, were evaluated. Sixty-three patients with ovarian cancer were followed up from 7 to 68 months. The positive expression rate of P-AKT and NF-kappaB p65 were higher in epithelial ovarian cancer than in normal ovarian tissue (P<0.01). Elevated P-AKT or NF-kappaB p65 expression was significantly correlated with late clinical stage (P<0.05 and P<0.01) and poor histological differentiation (both P<0.01). P-AKT expression was significantly correlated with NF-kappaB p65 immunostaining (phi=0.272, P<0.05). Elevated expression of P-AKT was negatively correlated with the survival of ovarian cancer patients, but it was not an independent prognostic factor after multivariate analysis. Overexpression of P-AKT and NF-kappaB p65 were involved in the carcinogenesis and metastasis of ovarian cancer. P-AKT might contribute to the malignant transformation through NF-kappaBp65 upregulation.

Diabetic ketoacidosis in pregnancy tends to occur at lower blood glucose levels: case-control study and a case report of euglycemic diabetic ketoacidosis in pregnancy.

BACKGROUND AND OBJECTIVE: The occurrence of diabetic ketoacidosis (DKA) during pregnancy is considered a medical emergency. The aims of the present study were to evaluate the incidence of DKA in pregnant and non-pregnant women with diabetes; to compare the blood glucose levels at the diagnosis of DKA in pregnant and non-pregnant women; and to show a case of euglycemic DKA in pregnancy. METHODS: The subjects consisted of 90 cases of DKA in pregnant women with diabetes and 286 cases of non-pregnant female inpatients receiving treatment for diabetes during 2001 to 2005 in our hospital. The incidence of DKA in pregnant and non-pregnant women with diabetes and the blood glucose levels at the diagnosis of DKA in pregnant and non-pregnant women were compared. RESULTS: DKA had a higher incidence in pregnant women with diabetes (8/90, 8.9%) than in non-pregnant women with diabetes (9/286, 3.1%) (P < 0.05). The blood glucose levels (mmol/L) in pregnant women with DKA were significantly lower than those in non-pregnant women with DKA (16.3 +/- 4.6 vs 27.5 +/- 4.8, P < 0.001). A case of euglycemic DKA in pregnancy was described whose serum glucose level was only 6.9 mmol/L. CONCLUSIONS: DKA in pregnant women with diabetes may occur more frequently, and at lower blood glucose levels than DKA in non-pregnant women with diabetes.

[Expression and significance of Edg4 and Edg7 in the placentas of patients with hypertensive disorder complicating pregnancy].

OBJECTIVE: To study Edg4 and Edg7 expression in placenta of women with hypertensive disorder complicating pregnancy, and to investigate the relation between the expression of lysophosphatidic acid (LPA) and hypertensive disorder complicating pregnancy. METHODS: Immunohistochemical SP method was used to measure the expressions of Edg4 and Edg7 in placenta of women with normal pregnancy, 20 women with gestational hypertension, 20 with mild preeclampsia, and with severe preeclampsia. RESULTS: (1) LOCATION: immunohistochemical staining for Edg4 and Edg7 protein were located at the membrane and endochylema of cytotrophoblast as well as decidua cells. (2) The positive expression of Edg4 protein and Edg7 protein on membrane and endochylema of cytotrophoblast was 25% and 20% (normal women), 60% and 40% (gestational hypertension), 80% and 65% (mild preeclampsia), and 83.3% and 86.7% (severe preeclampsia). The expression of Edg4 and Edg7 protein in mild preeclampsia and severe preeclampsia was significantly correlated with the degree of differentiation (P < 0.05). The expression of Edg4 and Edg7 protein showed an insignificant difference in normal pregnant women and gestational hypertension (P > 0.05). (3) The positive expression of Edg4 protein and Edg7 protein on membrane and endochylema of decidua was 20% and 25% (normal pregnancy), 55% and 50% (gestational hypertension), 70% and 55% (mild preeclampsia), and 83.3% and 73.3% (severe preeclampsia) respectively. The expression of Edg4 and Edg7 protein in mild preeclampsia and severe preeclampsia showed a significant correlation with the degree of differentiation (P < 0.05). The expression of Edg4 and Edg7 protein showed an insignificant difference in normal pregnancy and gestational hypertension (P > 0.05). CONCLUSIONS: The high expression of Edg4 and Edg7 protein in the placentas of patients with hypertensive disorder complicating pregnancy indicates that LPA combines with Edg4 and Edg7, inducing the occurrence of hypertensive disorder complicating pregnancy.

[Vector construction and silencing effect of Edg4 gene targeted small interfering RNA in ovarian cancer cell line].

OBJECTIVE: To construct the recombinant eukaryotic expression vector pRNAT-U6.1-siEdg4 which carries small interfering RNA (siRNA) of Edg4 and observe the silencing effect of Edg4 gene targeted siRNA in ovarian cancer cell line SKOV3. METHODS: The Edg4 gene-targeted hairpin siRNA sequence was designed according to the Edg4 sequence in Genbank, and the two complementary oligo nucleotide strands were synthesized and annealed and inserted into the pRNAT-U6.1 plasmid to build a recombinant Edg4 siRNA eukaryotic expression vector, which was sequenced and identified to contain the correct Edg4 siRNA sequence. The human ovarian carcinoma cell lines SKOV3 were transfected with the vector using lipofectamine method. The efficiency of transfecting cells was observed with fluorescent microscope and the mRNA expression level of Edg4 gene was detected by real time quantitative PCR. The LPA levels in cell supernatants were detected using a biochemical method. And the apoptosis of SKOV3 cells induced by the vector was evaluated by flow cytometry. RESULTS: The recombinant eukaryotic expression vector was confirmed to contain correct Edg4 siRNA sequence by PCR and sequencing. After transfection large amounts of green fluorescence were seen in plasma and nuclei of SKOV3 cells and the positive cell rates were 64%. The expression level of Edg4 mRNA in transfected SKOV3 cell line was significantly decreased (0.05 +/- 0.01vs 0.29 +/- 0.04, P < 0.05). The decrease in LPA level in the cell supernatants was revealed [(3.0 +/- 1.0) vs (7.5 +/- 2.2)micromol/L, P < 0.05]. The apoptosis rate of transfected SKOV3 was increased obviously (53.38% vs 0.51%, P < 0.05). CONCLUSIONS: We have successfully constructed the recombinant eukaryotic expression vector containing Edg4 gene targeted siRNA (pRNAT-U6.1-siEdg4). The vector could effectively transfect SKOV3 cell line, and obviously suppress the Edg4 mRNA expression and induce cell apoptosis in ovarian cancer cell line SKOV3.