Role of ADAM and ADAMTS disintegrin and metalloproteinases in normal pregnancy and preeclampsia.

Normal pregnancy (NP) involves intricate processes starting with egg fertilization, proceeding to embryo implantation, placentation and gestation, and culminating in parturition. These pregnancy-related processes require marked uteroplacental and vascular remodeling by proteolytic enzymes and metalloproteinases. A disintegrin and metalloproteinase (ADAM) and ADAM with thrombospondin motifs (ADAMTS) are members of the zinc-dependent family of proteinases with highly conserved protein structure and sequence homology, which include a pro-domain, and a metalloproteinase, disintegrin and cysteine-rich domain. In NP, ADAMs and ADAMTS regulate sperm-egg fusion, embryo implantation, trophoblast invasion, placental angiogenesis and spiral arteries remodeling through their ectodomain proteolysis of cell surface cytokines, cadherins and growth factors as well as their adhesion with integrins and cell-cell junction proteins. Preeclampsia (PE) is a serious complication of pregnancy characterized by new-onset hypertension (HTN) in pregnancy (HTN-Preg) at or after 20 weeks of gestation, with or without proteinuria. Insufficient trophoblast invasion of the uterine wall, inadequate expansive remodeling of the spiral arteries, reduced uteroplacental perfusion pressure, and placental ischemia/hypoxia are major initiating events in the pathogenesis of PE. Placental ischemia/hypoxia increase the release of reactive oxygen species (ROS), which lead to aberrant expression/activity of certain ADAMs and ADAMTS. In PE, abnormal expression/activity of specific ADAMs and ADAMTS that function as proteolytic sheddases could alter proangiogenic and growth factors, and promote the release of antiangiogenic factors and inflammatory cytokines into the placenta and maternal circulation leading to generalized inflammation, endothelial cell injury and HTN-Preg, renal injury and proteinuria, and further decreases in uteroplacental blood flow, exaggeration of placental ischemia, and consequently fetal growth restriction. Identifying the role of ADAMs and ADAMTS in NP and PE has led to a better understanding of the underlying molecular and vascular pathways, and advanced the potential for novel biomarkers for prediction and early detection, and new approaches for the management of PE.

Predicting Histological Healing and Recurrence in Ulcerative Colitis by Assessing Mucosal Vascular Pattern Under Narrow-Band Imaging Endoscopy.

This study investigated the predictive value of narrow-band imaging (NBI) endoscopic staging of different mucosal vascular patterns (MVPs) in patients with ulcerative colitis (UC) for histological healing or clinical recurrence of patients with UC. A total of 124 patients with UC in clinical remission attending the First Affiliated Hospital of Weifang Medical College were included in the study and underwent NBI colonoscopy. Inflammatory activity was assessed in the intestine using the Mayo endoscopic score (MES) and the MVP. Mucosal inflammation was histologically graded using the Nancy index (NI). The colons of 124 patients with UC were staged according to NBI endoscopic MVP staging criteria. The differences between NBI colonoscopy MVP typing and white light endoscopic MES in assessing histological healing (HH) were statistically significant (p < 0.001), and there was a moderate correlation between MES and the degree of HH (r = 0.471, p < 0.001). In addition, there was a significant correlation between the severity of mucosal activity determined by white light endoscopy (WLE) and MVP staging (r = 0.811, p < 0.001). The differences between NBI endoscopic MVP staging and white light endoscopic MES in assessing UC recurrence were statistically significant (p < 0.001). Spearman's correlation analysis showed a moderate correlation between NBI endoscopic MVP staging and clinical recurrence. NBI endoscopic MVP staging can predict HH and clinical recurrence status better than WLE.

Relationship between Human Papillomavirus Prevalence and DNA Damage in Cervical Cancer Population in Gansu Province, China.

OBJECTIVES: The aim of this study was to assess the possible reason of the high incidence and mortality of cervical cancer in Longnan, China. MATERIALS AND METHODS: 147 and 124 invasive squamous-cell carcinoma (SCC) samples from Longnan and different cities and districts of Gansu province were collected in the present study. All the samples were obtained from patients who underwent biopsies with colposcopy or advanced operations and were evaluated by experienced pathologists. HPV genotypes were examined with a validated HPV subtypes kit. The prevalence of HPV infection in SCC patients of China was analyzed by evidence-based medicine in the published literature. The markers of DNA damage response (DDR) - ATMpSer1981, H2AXp Ser139 (γH2AX), Chk2pThr68, and p53 - were analyzed by immunohistochemistry. RESULTS: HPV positivity, high-risk and multiple HPV positivity, and HPV58 infection were significantly higher in Longnan. Our results show that the prevalence of HPV infection in SCC patients of Longnan are consistent with the HPV prevalence in China. ATM, γH2AX, and p53 expressions in total and HPV+ samples were also higher in Longnan. CONCLUSIONS: HPV-related DDR activation may be one reason for the high incidence and mortality of Longnan cervical cancer.

MiR-95-5p involves in the migration and invasion of trophoblast cells by targeting low density lipoprotein receptor-related protein 6.

AIMS: Low density lipoprotein receptor-related protein 6 (LRP6) has been demonstrated to control trophoblast cell invasion, but its regulatory gene remains undefined. In this study, microRNA (miR) regulating LRP6 were explored to elucidate the potential mechanism of preeclampsia (PE). METHODS: Firstly, the expression of LRP6 in PE tissues was detected by immunohistochemical staining and quantitative real-time polymerase chain reaction (qRT-PCR) assay. Prediction software predicted that LRP6 might be the target gene of miR-95-5p, and verified by double-luciferase reporter analysis. qRT-PCR assay measured the expression of miR-95-5p in PE tissues and trophoblast cell lines. Then, we transfected miR-95-5p mimic, inhibitor, LRP6, or mimic plus LRP6 into trophoblast cell lines, and analyzed their influences on cell migration and invasion by wound healing and Transwell experiments. The expressions of matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitors of metalloproteinase (TIMP)-1 in transfected cells were examined by western blot (WB) analysis. RESULTS: LRP6 was low-expressed in PE tissues, while miR-95-5p expression was high-expressed. MiR-95-5p negatively regulated the LRP6 expression in trophoblast cells. Both up-regulated LRP6 and down-regulated miR-95-5p can not only promote the migration and invasion of trophoblast cells, but also raised the expressions of MMP-2 and MMP-9 and inhibited the expression of TIMP-1. The over-expression of miR-95-5p suppressed the metastasis of trophoblast cells and rescued LRP6-induced increase of MMP-2 and MMP-9 and reduction of TIMP-1. CONCLUSION: MiR-95-5p involved in the migration and invasion of trophoblast cells by targeting LRP6, which might be a potential therapeutic target for PE.

HIF­3α affects preeclampsia development by regulating EVT growth via activation of the Flt­1/JAK/STAT signaling pathway in hypoxia.

Preeclampsia (PE) is a common obstetric disease occurring after 20 weeks of gestation. Hypoxia­inducible factor (HIF)­3α potentially functions as a regulatory factor in PE development, however its specific molecular mechanism remains to be elucidated. The present study aimed to investigate the function of HIF­3α in trophoblast cell line HTR­8/SVneo, to provide a better understanding of the pathology and treatment of PE. Normal and PE placentas were obtained from pregnant women. HTR8/SVneo cells were cultured under the condition of normoxia or hypoxia, pretreated with or without AG490, then transfected with HIF­3α. The gene expression levels of HIF­3α and Fms like tyrosine kinase receptor (Flt) 1 extracted from the placentas and cells were detected by reverse transcription­quantitative PCR, and the expression levels of proteins and Janus kinase signal transducer and activator of transcription (JAK/STAT) phosphorylation were detected by western blot analysis. Viability and apoptosis of the treated cells were assessed by MTT and flow cytometry. The results demonstrated that HIF­3α and Flt­1 gene expression levels of PE placentas were reduced compared with normal placentas. Under a hypoxic environment, the expression levels of HIF­3α and Flt­1, the phosphorylation of JAK/STAT and the cell viability of HTR8/SVneo cells were increased at first and then reduced, whereas cell apoptosis was promoted over time. Under chronic hypoxia, the expression levels of HIF­3α and Flt­1, JAK/STAT pathway phosphorylation and cell viability of AG490­treated HTR8/SVneo cells were reduced, but cell apoptosis was promoted. However, the upregulation of HIF­3α in HTR8/SVneo cells markedly reversed the effects of AG490 on the cells under hypoxia. Thus, the present study preliminarily demonstrated that HIF­3α was involved in PE development by regulating extravillous cytotrophoblast growth via Flt­1 and the JAK/STAT signaling pathway.

The relationship between iron metabolism, stress hormones, and insulin resistance in gestational diabetes mellitus.

AIM: To analyze the relationship between iron metabolism index and stress hormones, insulin resistance, and oxidative stress in gestational diabetes mellitus (GDM). METHODS: From January to November 2019, 75 patients with GDM were selected as GDM group, according to age of 1:1; 75 normal pregnant women were selected as Control group. Blood glucose, insulin, stress hormones such as cortisol, norepinephrine (NE), and epinephrine (E), and iron metabolism index such as serum iron, serum ferritin (SF), and transferrin saturation (TS) were measured. Insulin resistance was evaluated by homeostasis model insulin resistance index (HOMA-IR). Multiple linear regression was used to analyze the relationship between iron metabolism index and stress hormones, insulin resistance, and oxidative stress. RESULTS: The levels of NE, E, serum iron, SF, and TS saturation in the GDM group were higher than Control group (t = 3.82, 2.75, 3.14, 6.12, and 3.90, P < 0.05, <0.05, <0.05, <0.01, <0.01); HOMA-IR was higher in the GDM group (t = 4.92, P < 0.01); malondialdehyde (MDA) was higher, while superoxide dismutase (SOD) was lower than Control group (t = 5.25, 4.98, both P < 0.01). Epinephrine, norepinephrine, cortisol, and serum ferritin were positively correlated (r = 0.21, 0.17, and 0.21); epinephrine, cortisol, and transferrin were positively correlated (r = 0.12, 0.31). There was a positive correlation between HOMA-IR and SF and TS (r = 0.34, 0.34). MDA was positively correlated with SF and TS (r = 0.24, 0.29); SOD was negatively related to SF and TS (r = -0.12, -0.17). CONCLUSIONS: Iron metabolism index is related to insulin resistance in GDM women. The change in iron metabolism may be involved in the pathogenesis of gestational diabetes caused by stress- adaptive disorder.

Stress adaptation is associated with insulin resistance in women with gestational diabetes mellitus.

AIM: Oxidative stress is known to increase the risk of insulin resistance (IR). The aim of this study was to investigate the association between stress hormones and IR in women with gestational diabetes mellitus (GDM), in an attempt to gain insights into the pathogenesis of GDM. METHODS: Recruited in this study were 70 GDM women and 70 healthy pregnant women as control. Malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), plasma epinephrine (E), noradrenaline (NE), glucagon, and cortisol levels were detected. IR was assessed by homeostasis model assessment of IR (HOMA-IR) in both groups. Correlations among stress hormones, oxidative stress, and IR were analyzed by Pearson's correlation after log transformation. RESULTS: Compared with the Control group, MDA was increased and anti-oxidative enzymes SOD and GSH were decreased significantly in the GDM group. Glucagon, E, and NE in the GDM group were increased by 22.42%, 36.82%, and 35.09%, respectively, as compared with those in the Control group. MDA showed a significant positive correlation, and SOD showed a negative correlation with HOMA-IR in the GDM group. In addition, HOMA-IR was positively related to glucagon, E, NE, and cortisol. CONCLUSIONS: Elevation of stress hormones and stress adaptation disturbance may be associated with the pathogenesis of GDM in pregnant women.

Reduced ELABELA expression attenuates trophoblast invasion through the PI3K/AKT/mTOR pathway in early onset preeclampsia.

INTRODUCTION: Early onset preeclampsia is linked to abnormal trophoblast invasion, leading to insufficient recasting of uterine spiral arteries and shallow placental implantation. This study investigated ELABELA (ELA) expression and its involvement in the pathogenesis of early onset preeclampsia. METHODS: We used immunohistochemistry, quantitative PCR and Western blot to calculate ELA levels in the placentas. Transwell assays were utilize to assess the invasion and migration of trophoblastic Cells. Western blot was used to identify the concentrations of vital kinases in PI3K/AKT/mTOR pathways and invasion-related proteins in trophoblast cells. RESULTS: ELA was expressed in villous cytotrophoblasts and syncytiotrophoblasts in placental tissue. Compared with the normal pregnancies, ELA mRNA and protein expression was significantly reduced in early onset preeclampsia placentas. In the HTR-8/SVneo cells, when ELA was knocked down, the invasion and migration capability of cells decreased significantly, with MMP2 and MMP9 expression downregulated and the expression of important kinases in the PI3K/AKT/mTOR pathways being significantly decreased compared to the control group. Overexpression of ELA was on the contrary. Besides, while PI3K was blocked, the invasion and migration capability of HTR-8/SVneo cells and the expression of key kinases in PI3K/AKT/mTOR pathways were decreased significantly. DISCUSSION: ELA stimulates the invasion and migration of trophoblastic cells through activation of downstream PI3K/AKT/mTOR pathway and is complicit in early onset preeclampsia pathogenesis. Our research offers a potential novel treatment for PE.

HPV infection associated DNA damage correlated with cervical precancerous lesions and cancer in the highest area of cervical cancer mortality, Longnan, China.

OBJECTIVES: This study was to assess whether human papillomavirus (HPV) resulting in genetic instability is one reason for the high incidence and mortality of cervical cancer in Longnan. METHODS: Between 2012 and 2016, a total of 346 samples from Longnan were collected and divided into four groups: cervicitis group (n=57), cervical intraepithelial neoplasia I group (CIN I, n=63), CIN II/III group (n=79) and invasive squamous cell carcinoma group (SCC, n=147). HPV E6/E7 mRNA was detected by Quantivirus® HPV E6/E7 RNA 3.0 assay (bDNA). The markers of DNA damage response (DDR) - ataxia telangiectasia mutated (ATM) pSer1981, H2AX pSer139 (γH2AX), Chk2 pThr68 and P53 - were analyzed by immunohistochemistry. RESULTS: The activation of ATM, γH2AX, Chk2 and P53 was increased with increasing severity of cervical lesion. A significant difference of ATM expression in simple infection was also shown accompanied by the cervical lesion. The expression of γH2AX between HPV16+ and HPV16- specimens, γH2AX and P53 between HPV58+ and HPV58- groups had statistical significance. The expression and copy number of HPV E6/7 mRNA increases with the cervical lesion severity. A significant difference was shown for P53 expression between HPV E6/7 mRNA+ and mRNA- specimens. A close correlation with CHK2 expression for HPV E6/7 mRNA+ and HPV16 E6/7 mRNA+ specimens and γH2AX and CHK2 expression for SCC specimens was shown between low and high viral load groups. CONCLUSIONS: DDR, HPV genotypes and HPV E6/E7 oncogene expression correlated with the level of dysplasia of cervical lesions. HPV infection resulted in genetic instability may be one reason for the high incidence and mortality in Longnan.

MiR-181c affects estrogen-dependent endometrial carcinoma cell growth by targeting PTEN.

MicroRNAs (miRNAs), which is a type of non-coding and single-stranded small molecule RNA, bind either completely or incompletely to 3'-UTR of the target gene mRNA to inhibit mRNA translation or degradation. In our study, we aimed to explore the roles and mechanisms of miR-181c in the apoptosis of RL95-2 human endometrial carcinoma cells. Cell activity and apoptosis were detected by cell counting Kit-8 (CCK-8) assay and flow cytometry (FCM), respectively. Related mRNAs and proteins expression was determined by quantitative real-time reverse transcription PCR (qRT-PCR) and western blot assays, respectively. The binding capacity of PTEN-3'-UTR and miR-181c was assessed by luciferase reporter assay. The obtained results suggested that E2 evidently increased the cell activity of RL95-2 cells. In addition, miR-181c inhibitor suppressed the cell viability and enhanced the apoptosis capacity of E2-induced RL95-2 cells and distinctly reduced the miR-181c expression. We also found that miR-181c could bind to PTEN-3'-UTR and miR-181c inhibitor up-regulated the expression level of PTEN in E2-induced RL95-2 cells. Besides, overexpression of PTEN markedly promoted the apoptosis of E2-induced RL95-2 cells through regulating the Bax and Bcl-2 expression, and modulated the expression of AKT pathway, p53 and Cyclin D. In conclusion, our findings revealed that miR-181c affected the estrogen-dependent endometrial carcinoma cell growth by targeting PTEN. The potential effects of miR-181c on the apoptosis of E2-induced RL95-2 cells suggest that miR-181c could be an effective target for endometrial carcinoma therapies.

Methylation of HIF3A promoter CpG islands contributes to insulin resistance in gestational diabetes mellitus.

BACKGROUND: Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance during pregnancy, and will lead to high risk of diabetes even after pregnancy. Hypoxia-inducible factor (HIF) family proteins are transcriptional factors that are highly correlated with methylation, which might be involved in the regulation of GDM. METHODS: Baseline clinical characteristics of the GDM patients and healthy women were analyzed. Omental tissue from GDM patients and control groups were collected and detected for the expression levels of HIF1A, HIF2A, and HIF3A. The CpG islands of HIF3A promoter were predicted by "methprimer" software, and the methylation level of CpG islands was detected by bisulfite sequencing PCR. RESULTS: HIF3A was downregulated in the omental tissue from GDM patients, whereas HIF1A and HIF2A were not affected. Furthermore, HIF3A expression was positively correlated with levels of estrogen receptor α (ESR1) and solute carrier family 2 member 4 (SLC2A4). Moreover, CpG islands of HIF3A promoter were highly methylated in GDM patients. In addition, methylation level of CpG islands could be upregulated by Estradiol (E2) treatment, since high dose of E2 reduced HIF3A mRNA expression in 3T3-L1 adipocytes. CONCLUSION: Our findings demonstrate that the expression level of HIF3A, but not HIF1A or HIF2A, is downregulated in GDM patients. The methylation status of HIF3A promoter region is highly correlated with GDM, which could be a novel therapeutic target for GDM treatment.

Overexpressed HO-1 is associated with reduced STAT3 activation in preeclampsia placenta and inhibits STAT3 phosphorylation in placental JEG-3 cells under hypoxia.

INTRODUCTION: Inadequate trophoblast invasion and placentation are widely believed to contribute to preeclampsia, and multiple lines of evidence indicate the involvement of hypoxia in preeclampsia. However, the molecular mechanisms underlying the association of placental hypoxia with preeclampsia are not clear. MATERIAL AND METHODS: The present study focused on the role in preeclampsia of heme oxygenase 1 (HO-1), which is an inducible isoform of HO in response to hypoxia, via examining the expression of HO-1 and the expression and phosphorylation (Tyr705) of Signal transducer and activator of transcription (STAT) 3 in preeclamptic placentas via the immunohistochemical method, western blotting assay and RT-qPCR method. Then we investigated the regulation by HO-1 of the expression and phosphorylation of STAT3 in human placental choriocarcinoma JEG-3 cells under hypoxia. RESULTS: There was upregulation of HO-1 at both mRNA (1.506 ±0.08347 (N = 37) vs. 1.000 ±0.08854 (N = 31), p < 0.0001) and protein (0.630 ±0.155 (N = 35) vs. 0.310 ±0.052, 0.630 ±0.155 (N = 35), p < 0.001) levels and a reduced level of STAT3 phosphorylation (Tyr 705) in the preeclamptic placental tissues, compared to normal placental tissues (0.143 ±0.027 (N = 35) vs. 0.194 ±0.028 (N = 35), p < 0.01). Also, in vitro experiments demonstrated that HO-1 was markedly promoted by hypoxia in human placental choriocarcinoma JEG-3 cells, 6 or 12 h post treatment (p < 0.05 or p < 0.01). However, the STAT3 phosphorylation (Tyr 705) was attenuated by sustained hypoxia (p < 0.01). Moreover, it was demonstrated that HO-1 overexpression significantly inhibited the hypoxia-promoted STAT3 phosphorylation (Tyr 705). CONCLUSIONS: HO-1 was overexpressed in PE placenta, in association with reduced STAT3 phosphorylation (Tyr 705). HO-1 inhibits the STAT3 phosphorylation in placental JEG-3 cells under hypoxia. Thus, we speculate that overexpressed HO-1 might contribute to the reduced STAT3 phosphorylation (Tyr 705) and the pathogenesis of preeclampsia.

[Establishing a Combination Model in Predicting Mortality of Lung Cancer].

OBJECTIVE: To identify a good combination model for predicting the mortality of lung cancer. METHODS: Mortality data of lung cancer from 2001-2013 were used to test three prediction model: dynamic series, exponential smoothing, and Joinpoint regression. Weight coefficients of the combination models were calculated using the arithmetic average method, the variance inverse method, the mean square error inverse method, and the simple weighted average method. RESULTS: The exponential smoothing model had the highest accuracy (79.67%) of prediction, followed by the Joinpoint linear model (74.27%). The combination of these two models resulted in better results. The arithmetic average method and the mean square error inverse method had the best prediction, with an accuracy of 86.87% and 85.80%, respectively. CONCLUSIONS: The combined model has higher accuracy than the single models in predicting the mortality of lung cancer.

Human papillomavirus genotypes associated with cervical precancerous lesions and cancer in the highest area of cervical cancer mortality, Longnan, China.

BACKGROUND: The mortality of cervical cancer in Longnan is as high as 39/10 million, ranking first in China. METHODS: Between 2012 to 2016, 329 samples with cervicitis, cervical intraepithelial neoplasia grade 1 to 3 (CINI to III), and invasive squamous cell carcinoma (SCC) were collected. HPV genotypes were examined with a validated kit for 23 different HPV subtypes. RESULTS: Compared to cervicitis, the HPV positivity is significantly higher in CINI, CIN II/III, and SCC (38.60%, 74.60%, 87.50% and 89.05%, P < 0.001) and the positivity is also higher in SCC compared to CINI (P < 0.01). The most frequently detected genotypes were HPV16 in cervicitis, HPV16, 58 and 52 in CINI and CIN II/III, and HPV16, 58 and 18 in SCC groups. HPV16 positivity in cervicitis, CINI, CIN II/III, and SCC patients were 45.46%, 46.81%, 60.32% and 78.69%, respectively. Compared to cervicitis and CINI, the odds ratios (OR) for SCC in HPV16 positive patients were 2.96 (95% confidence interval [CI]: 1.09-8.00, P < 0.05) and 4.20 (95% confidence interval [CI]: 2.05-8.61, P < 0.001), respectively. In addition, the multiple infections in cervicitis, CINI, CINII/III and SCC group are 9.09%, 27.66%, 26.98% and 25.41% and HPV16 + 58 was the most common combinations. CONCLUSION: These findings highlight the key role of HPV16, 58, 52 and 18 in the development of CIN and SCC in Longnan women and a fully aware of regional differences in HPV genotype distribution are tasks for cervical cancer control and prevention.

Tumor suppressor in lung cancer 1 gene expression in epithelial ovarian cancer.

OBJECTIVES: Loss of Tumor Suppressor in Lung Cancer 1(TSLC1) was observed in many different cancers, but there were only limited research on TSLC1 gene and its roles in cancer suppression in ovarian cancer. This study explores the relationship between TSLC1 gene expression and the ovarian epithelial cancer. MATERIALS AND METHODS: Expression levels of TSLC1 were detected by immunohistochemical staining on formal-fixed paraffin embedded pathological specimen. A total of 259 samples were collected, including 24 benign ovarian tumor and 235 malignant ovarian cancers among them. RESULTS: Suppressed expression of TSLC1 protein was observed in 87.8% of poorly differentiated, 85.1% of moderately differentiated, and 46% of well differentiated ovarian epithelial cancers, respectively. There were none suppressed TSLC1 expression in benign ovarian tumor. Kruskal-Wallis test showed a significant association between the expression levels of TSLC1 gene and the degree of ovarian cancer differentiation (P < 0.001). CONCLUSIONS: Decreased expression of TSLC1 is associated with the differentiation in ovarian epithelial cancer. TSLC1 might be used as a molecular marker of severity in early stage ovarian cancer, and to help differentiating benign and malignant ovarian tumors.

Kruppel-like factor 4 (KLF-4) inhibits the epithelial-to-mesenchymal transition and proliferation of human endometrial carcinoma cells.

Kruppel-like factors (KLFs) are a group of transcriptional regulators, being tumor-suppressive in various types of cancers, but not clear in human endometrial carcinoma (EC). We investigated the KLF-4 expression in both mRNA and protein levels in 29 EC specimens with RT-qPCR and Western blotting methods, and then to determine its promotion to Epithelial-to-mesenchymal transition (EMT) and proliferation of EC Ishikawa cells, via analyzing EMT-associated markers and via CCK-8 and colony forming assay. We found the downregulation of KLF-4 in the 29 EC specimens, correlating with the EC malignance. Moreover, we confirmed reduced levels of EMT and cell proliferation of Ishikawa cells post-KLF-4 overexpression. In conclusion, the significantly reduced KLF-4 correlated with the EC malignance. And the overexpressed KLF-4 promoted the EMT and proliferation of EC cells in vitro. The present study recognized the tumor suppressive role of KLF-4 in EC.

[Cancer burden in the Jinchang cohort].

OBJECTIVE: To understand the disease burden caused by cancers in Jinchang cohort, and develop effective strategies for cancer prevention and control in this population. METHODS: The cancer mortality data from 2001 to 2013 and the medical records for cancer patients from 2001 to 2010 in Jinchang cohort were collected. The disease burden caused by cancer was analyzed by using mortality rate, potential years of life lost (PYLL), working PYLL (WPYLL), and direct economic burden. RESULTS: During 2001-2013, in Jinchang cohort, the five leading cancers ranked by mortality rate were lung cancer (78.06/100,000), gastric cancer (38.03/100,000), liver cancer (37.23/100,000), esophageal cancer (19.06/100,000), and colorectal cancer (9.53/100,000). The five leading cancers in terms of PYLL (person-years) and WPYLL (person-years) were lung cancer (3480.33, 1161.00), liver cancer (2809.03, 1475.00), gastric cancer (2120.54, 844.00), esophageal cancer (949.61, 315.00), and colorectal cancer (539.90, 246.00). From 2001 to 2010, the five leading cancers in term of average daily cost of hospitalization were gastric cancer (8,102.23 Yuan), esophageal cancer (7135.79 Yuan), colorectal cancer (7064.38 Yuan), breast cancer (6723.53 Yuan), and lung cancer (6309.39 Yuan). CONCLUSIONS: The cancers common causing higher disease burden in Jinchang cohort were lung cancer, gastric cancer, liver cancer, esophageal cancer and colorectal cancer. The lung cancer disease burden was the highest.

Rapid detection of active human cytomegalovirus infection in pregnancy using loop-mediated isothermal amplification.

Understanding the association between congenital human cytomegalovirus (HCMV) infection and active maternal HCMV infection during pregnancy is important for maternal and neonatal healthcare. In the present study, a loop-mediated isothermal amplification (LAMP) method was established for the detection of CMV DNA from whole blood or amniotic fluid samples, using reverse transcription-quantitative polymerase chain reaction. The results of the present study demonstrated that the CMV LAMP assay detection was specific for CMV DNA, whereas it did not detect viral DNA from herpes simplex type 1 (HSV-1), HSV-2, varicella zoster virus, HSV-6 or HSV-7. Sensitivity determination using serially-diluted CMV glycoprotein B-containing plasmids, demonstrated that >10 copies per tube were detectable using the CMV LAMP method. Furthermore, the detection results, using the LAMP method for 336 whole blood samples, demonstrated that at a threshold of 10(1)-10(4) copies per tube, the sensitivity of this method was 86.96-100%, the specificity was 97.24-100%, the positive predictive value was 76.92-100% and the negative predictive value was 99.05-100%. The results for 11 amniotic fluid samples from pregnant women with whole blood CMV-positive and 15 control amniotic fluid samples, indicated that the CMV LAMP assay was sensitive and specific for CMV detection. In conclusion, in the present study, a CMV LAMP method was developed, which was shown to be sensitive, specific and efficient in the detection of HCMV infection. Furthermore, CMV LAMP is capable of detecting active CMV infection in pregnant women. Therefore, the current study provides novel insights into diagnostic approaches for active CMV infection in pregnant women.

Metformin inhibits angiogenesis induced by interaction of hepatocellular carcinoma with hepatic stellate cells.

Accumulated evidences indicate metformin is associated with reduced risk of hepatocellular carcinoma (HCC) in diabetic patients, which inspired researchers to explore its therapeutic potentials in HCC. Since Hepatic stellate cells (HSCs) are believed to be the key contributors to tumor microenvironment in HCC and promotes tumor development, here, we explored the effect of metformin on tumor angiogenesis induced by interplay of HCC and HSCs. Our results showed that conditional medium from co-culture of HCC/HSCs induced VEGF secretions and stimulated human umbilical vein endothelial cells (HUVEC) tube formation. However, 25 µM metformin could inhibit this angiogenesis effect. Furthermore, knockdown AMPK of HSCs, not HCC, could abolish inhibition caused by metformin. Our finding suggested that metformin could inhibit HCC angiogenesis through targeting on HSCs through AMPK pathway.

Diethyl 4,5-diphenyl-3,6-bis-(trimethyl-sil-yl)benzene-1,2-dicarboxyl-ate.

In the title compound, C(30)H(38)O(4)Si(2), the two phenyl rings are twisted away from the central benzene ring by 70.28 (8) and 67.42 (7)°. The two Si atoms attached to the benzene ring deviate in opposite directions from the ring plane by 0.258 (3) and 0.206 (3) Å, respectively. One ethyl group is disordered over two conformations in a 0.568 (5):0.432 (5) ratio. The crystal packing exhibits weak inter-molecular C-H⋯O inter-actions.