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.

Integrated analysis of key microRNAs /TFs /mRNAs/ in HPV-positive cervical cancer based on microRNA sequencing and bioinformatics analysis.

BACKGROUND: Cervical squamous cell carcinoma (CESC) is one of the most common malignancies associated with mortality in females. Its onset and prognosis are primarily concerned with persistent infection with high-risk types of human papillomavirus (HPV). However, the molecular mechanisms of HPV-positive CESC remain unclear. METHODS: In this study, we conducted a high-throughput sequencing to identify differentially expressed miRNAs (DEMs). Besides, three series were selected from the Gene Expression Omnibus (GEO) database to identify differentially expressed genes (DEGs). Then the miRNA-TF-gene regulatory network was constructed using bioinformatic methods. Genes in the network were performed functional enrichment analysis and protein-protein interaction (PPI) network analysis. Ultimately, the expression levels of six key miRNAs, TFs, and mRNAs were validated by 20 HPV-positive CESC tissues and 15 normal cervical samples. RESULTS: A total of 52 DEMs and 300 DEGs differed between the HPV-positive CESC and normal cervical samples. Then the miRNA-TF-gene regulatory network was constructed consisting of 22 miRNAs, 6 TFs, and 76 corresponding genes, among which miR-149-5p, miRNA-1248 and E2F4 acted as key regulators. PPI network analysis showed that ten genes including TOP2A, AURKA, CHEK1, KIF11, MCM4, MKI67, DTL, FOXM1, SMC4, and FBXO5 were recognized as hub genes with the highest connectivity degrees. Besides, five key molecules miRNA-149-5p, E2F4, KIF11, DTL, and SMC4 were suggested to play crucial roles in the development of HPV-positive CESC. CONCLUSION: These results present a unique insight into the pathological mechanisms of HPV-positive CESC and possibly provides potential therapeutic targets.

Potential protein biomarkers for systemic lupus erythematosus determined by bioinformatics analysis.

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disorder, and its pathogenesis in males and in cases without accompanying lupus nephritis (LN-) is not fully understood. In this study, we identified 90 (82 up- and 8 downregulated) differentially expressed genes (DEGs) common to female LN-, female LN+ and male LN+ using the GSE65391 and GSE49454 gene expression datasets from Gene Expression Omnibus database (GEO). The protein-protein interaction (PPI) network of 70 DEGs was constructed using STRING and cytoscape, and the Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the PPI network was significantly enriched in defense response to virus, cytosol, protein binding and measles. Sixteen hubgenes were identified from this PPI network, and Literature Mining Gene Networks molecular of GenCLiP 2.0 showed strong interaction between STAT1, DDX58 and IFIT1. Enrichment analysis of hubgenes in published literature showed the involvement of immune response and interferon-related genes in the pathogenesis of SLE. In addition, the transcription factors STAT1 & 2 and IRF6 & 9 had high Normalized Enrichment Score (NES). The 70 DEGs with PPI network and 16 hubgenes are potential biomarkers of SLE, and can help improve diagnosis and develop individualized therapies.

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.

Using a panel of multiple tumor-associated antigens to enhance the autoantibody detection in the immunodiagnosis of ovarian cancer.

BACKGROUND: Ovarian cancer (OC) is a major malignancy affecting a large population over the world, and a biomarker that holds diagnostic potential is of critical importance. Recently, autoantibodies have been indicated as biomarkers in multiple cancer research. The current study was designed to explore the practice of using autoantibodies in diagnostic settings by the enzyme-linked immunosorbent assay of sera with a panel of tumor-associated antigens (TAAs). METHODS: A panel of 12 TAAs was selected to detect the corresponding autoantibodies in sera sampled from 132 OC patients as case group and 147 normal healthy individuals as the control group. The diagnostic potential of this panel was evaluated by conventional evaluation, receiver operating characteristic (ROC) curve analyses, and classification tree analysis. RESULTS: When the cutoff values were set as mean ± 2 SD for normal healthy individuals, the positive rates of antibodies to any single TAA were less than 20% both in OC and in normal healthy individuals. In a parallel screening approach, a panel of nine TAAs (p53, C-myc, p90, p62, AHSG, 14-3-3zeta, RalA, Koc, and p16), obtained optimal diagnostic performance in OC with the sensitivity of 61.4% at the 85.0% specificity. In addition, when the nine TAAs were combined with CA125, the sensitivity and specificity were improved to 94.7% and 78.2%, respectively. The ROC curve analyses showed that only the area under the receiver operating characteristic curves (AUCs) of antibodies against C-myc, Koc, and RalA was beyond 0.6, which were 0.732, 0.668, and 0.665, respectively. The AUC of the combination was up to 0.914 (P < 0.05). Decision tree analysis showed that C-myc, HCC1.3, RalA, and CA125 held high potential in the detection of OC. The panel of nine TAAs also identified 78.8% of OC patients who had normal CA125 levels in their serum samples, indicating that elevated CA125 and anti-TAA antibodies appeared to be independent but supplementary biomarkers for diagnosing OC. CONCLUSIONS: In summary, the current study further supports that a customized TAA panel can serve as a promising and powerful tool for immunodiagnosis of OC and may be particularly useful in patients with normal CA125 levels.

MicroRNA-155 Suppresses Mesangial Cell Proliferation and TGF-ß1 Production via Inhibiting CXCR5-ERK Signaling Pathway in Lupus Nephritis.

Increasing evidence shows miR-155 plays an important role in regulating inflammatory processes in systemic lupus erythematosus (SLE), especially in lupus nephritis (LN). Because the chemokine CXCL13 is implicated in the pathogenesis of LN, here we examined whether miR-155 can modulate the activity of CXCL13 or its receptor CXCR5. We determined the expression of CXCL13 in normal and MRL/lpr mice and found elevated levels of CXCL13 in the kidneys of MRL/lpr mice compared with normal kidneys. Besides, CXCL13 expression was mainly detected in the glomerulus, specifically to mesangial areas. We then transfected a miR-155 mimic in human renal mesangial cells (HRMCs) to overexpress miR-155 and detected decreased protein levels of CXCR5 by western blot analysis. Transfection of the miR-155 mimic into CXCL13-treated HRMCs resulted in a significantly reduced proliferation rate of HRMCs as measured by the cell-counting assay and flow cytometry. Moreover, increased intracellular miR-155 also led to decreased phosphorylation of ERK and TGF-ß1 production. Together, these results revealed that miR-155 may play a role in the pathogenesis of LN.

Effect of metformin use on the risk and prognosis of endometrial cancer: a systematic review and meta-analysis.

BACKGROUND: Previous studies have suggested that metformin may be useful for preventing and treating endometrial cancer (EC), while the results have been inconsistent. This systematic review and meta-analysis aimed to investigate the association between metformin use and risk and prognosis of patients with EC. METHODS: PubMed, Embase, and the Cochrane Library databases were searched for observational studies evaluating the effect of metformin on EC prevention or treatment. The odds ratio (OR) was used for analyzing risks, and the hazard ratio (HR) was used for analyzing survival outcomes. A random-effects model was used for data analysis. RESULTS: Seven studies reported data on EC risk. The pooled results suggested that metformin was not significantly associated with a lower risk of EC [OR = 1.05, 95% confidence interval (CI) 0.82-1.35, P = 0.70]. For patients with diabetes, metformin showed no advantage in reducing the EC risk compared with other interventions (OR = 0.99, 95% CI 0.78-1.26, P = 0.95). Further, seven studies were included for survival analysis. The pooled data showed that metformin could significantly improve the overall survival of patients with EC (HR = 0.61, 95% CI 0.48-0.77, P < 0.05) and reduce the risk of EC recurrence (OR = 0.50, 95% CI 0.28-0.92, P < 0.05) Finally, we noted metformin was associated with significantly improving the overall survival of EC patients among diabetes (HR = 0.47; 95%CI 0.33-0.67, P < 0.05). CONCLUSIONS: This meta-analysis did not prove that metformin was beneficial for preventing EC. However, metformin could prolong the overall survival of patients with EC and reduce their risk of cancer relapse.

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.

CXCL13 Promotes Proliferation of Mesangial Cells by Combination with CXCR5 in SLE.

As a CXC subtype member of the chemokine superfamily, CXCL13 is considered to be involved in systemic lupus erythematosus (SLE), especially in lupus nephritis (LN). To determine the effect of CXCL13 on SLE and explore the potential mechanisms, we tested serum concentrations of CXCL13 in patients and healthy individuals and found that CXCL13 expression was high in SLE patients especially in LN patients. When we treated human renal mesangial cells (HRMCs) in vitro with recombinant human CXCL13, the cell proliferation was accelerated, which was tested by Cell Counting Kit-8 assay and flow cytometry. Western blot and immunofluorescence assay revealed that CXCL13 would lead to phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, the effect was weakened after the silence of CXCR5. The results of our study elaborated that high expression of CXCL13 could be involved in the pathogenesis of LN.

Comparison of Two Techniques of Laparoscopy-Assisted Peritoneal Vaginoplasty.

Neovagina creation is essential for patients with the Mayer-Rokitansky-Kuster-Hauser syndrome. We compared a technique involved the pushing down of the peritoneum with the technique of separating the peritoneum for laparoscopy-assisted peritoneal vaginoplasty. We collected patients with congenital absence of vagina who underwent laparoscopy-assisted peritoneal vaginoplasty of the First Affiliated Hospital of Zhengzhou University between January 2011 and May 2013. The 2 surgical groups (pushing group and separating group) were compared for various parameters. The values of the following parameters were significantly lower for the pushing group compared with the separating group: mean operating time (78 ± 13 minutes vs 135 ± 28 minutes), mean duration of hospitalization (12.9 ± 2.7 days vs 18.0 ± 3.8 days), mean cost of hospitalization (14 016 ± 1640 RMB vs 18 783 ± 2143 RMB), requirement for a drainage tube (4% vs 27%; χ(2) = 8.864), requirement for analgesic drugs (20% vs 40%; χ(2) = 3.977), and postoperative rehospitalization (3.3% vs 10.0% at 2 months and 6.7% vs 26.7% at 6 months; χ(2) = 4.268 and 5.196). Mean values for blood loss (57 ± 19 mL vs 66 ± 20 mL), time to pass gas (21 ± 4 hours vs 23 ± 7 hours), and length of the reconstructed vagina (9.0 ± 0.4 cm vs 8.9 ± 0.5 cm) were not significantly different between the 2 groups. In addition, mean postoperative Female Sexual Function Index score did not differ significantly between the 2 groups or among the 2 groups and a control group (27.0 ± 4.8 vs 26.7 ± 5.2 vs 27.9 ± 4.5; p > .05). The technique involving pushing down of the peritoneum offers advantages of reduced cost, complications, hospitalization, operative time, and pain over the traditional technique. Sexuality approaches so-called "normal" sexuality.

Efficacy and safety of cord blood-derived cytokine-induced killer cells in treatment of patients with malignancies.

BACKGROUND AIMS: Adoptive immunotherapy with the use of cytokine-induced killer (CIK) cells represents an effective therapeutic option for treating malignancies. The characteristics and function of cord blood-derived CIK (CB-CIK) cells have been evaluated both in vitro and in vivo. In this study, we assessed the efficacy and safety of administering CB-CIK cells to patients with cancer. METHODS: In this retrospective clinical trial, 15 patients with cancer received CB-CIK therapy with different cycles from April 2012 to August 2014. CB-CIK cells demonstrated a high percentage of main functional fraction CD3(+)CD56(+) and efficient anti-tumor activity in vitro. RESULTS: After the infusion of CB-CIK cells, the subsets of CD3(+)CD4(+) T lymphocytes and CD3(-)CD56(+) T cells in the peripheral blood were significantly increased compared with those before the therapy. Of 15 patients, one patient with hepatocellular cancer and one patient with esophageal cancer achieved complete responses, two patients with ovarian cancer obtained partial remissions, 10 patients had stable disease and one patient with hepatocellular cancer had progressive disease. Acute toxicities including fever, slight fever, dizziness and other neurologic toxicities were few and occurred in patients after infusion of CB-CIK cells. CONCLUSIONS: These results demonstrated the feasibility and safety of treating malignancies with CB-CIK cells. The study provides a potential therapeutic approach for the patients with poor health or older patients who cannot tolerate repeated collection of blood.

Humoral autoimmune responses to insulin-like growth factor II mRNA-binding proteins IMP1 and p62/IMP2 in ovarian cancer.

Ovarian cancer is one of the leading causes of cancer-related deaths among women. There is an urgent need of better approaches for the identification of appropriate biomarkers in the early detection of ovarian cancer. The aim of this study was to elucidate the significance of autoantibodies against insulin-like growth factor II mRNA-binding proteins (IMPs) in patients with ovarian cancer. In this study, autoantibody responses to two members (IMP1 and p62/IMP2) of IMPs were evaluated by enzyme-linked immunosorbent assay (ELISA), western blotting, and indirect immunofluorescence assay in sera from patients with ovarian cancer and normal human individuals. The results have demonstrated that both IMP1 and p62/IMP2 can induce relatively higher frequency of autoantibody responses in patients with ovarian cancer (26.5% and 29.4%) compared to normal individuals (P<0.01). Our preliminary data suggest that IMP1 and p62/IMP2 can stimulate autoimmune responses in ovarian cancer, and anti-IMP1 and anti-p62/IMP2 autoantibodies could be used as potential biomarkers in immunodiagnosis of ovarian cancer.

[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.

The G protein-coupled receptor GPR30 mediates the nontranscriptional effect of estrogen on the activation of PI3K/Akt pathway in endometrial cancer cells.

OBJECTIVE: The goal of this study was to investigate the effect of G protein-coupled receptor 30 (GPR30) on the activation of PI3K/Akt pathway induced by E2 in endometrial cancer cells. METHODS AND MATERIALS: Immunohistochemistry was performed to determine the location and expression of GPR30, estrogen receptors (ERs), Akt, and phosphorylated Akt. We also investigated the expression of GPR30, ERs, and the level of phosphorylation of Akt induced by E2 in endometrial cancer cells, Ishikawa cells, and HEC-1A cells. We down-regulated the expression of GPR30 in endometrial cancer cell lines by transfection with shGPR30-pGFP-V-RS, a GPR30 antisense expression vector. The cells were then subjected to a proliferation assay. Immunoprecipitation assay was performed to determine whether GPR30 directly bind to PI3K. The stable transfected cells resuspension of 100 µL (5 × 10(6) cells) was injected subcutaneously into the right flank of athymic mice to perform xenograft tumor formation assays. RESULTS: E2 stimulated cell proliferation and induced GPR30 expression and PI3K/Akt pathway activation in endometrial cancer cells, Ishikawa cells, and HEC-1A cells, whereas the expression of ERs remained unchangeable. Down-regulation of GPR30 decreased the phosphorylation of Akt and reduced cell proliferation, and GPR30 did not bind to PI3K. Down-regulation of GPR30 significantly inhibited the tumor growth of HEC-1A cells in athymic nude mice. CONCLUSIONS: These findings suggest that GPR30 mediates the nontranscriptional effect of estrogen on the activation of PI3K/Akt pathway in endometrial cancer cells.

[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.