No abstract

Tumor invasion and metastasis are complex biological processes. Matriptase and its endogenous inhibitor, hepatocyte growth factor activator inhibitor-1 (HAI-1) are involved in invasion and metastasis. To evaluate the ratio of matriptase/HAI-1 and their potential therapeutic value in ovarian cancer, HO-8910 human ovarian cancer cells and the homologous high-metastatic HO-8910PM cells were used as in vitro cellular models ovarian cancer. The invasive and metastatic abilities, and the expression of matriptase and HAI-1 in these cells were detected using scratch assays, Transwell chamber assays, reverse transcription-quantitative polymerase chain reaction, western blotting and fluorescent immunocytochemistry. Following infection with lentivirus-mediated matriptase-targeting small interfering RNA (siRNA), cell cycle progression and apoptosis were also analyzed. The migration distance and number of invading HO-8910PM cells were significantly increased compared with HO-8910 cells. HO-8910PM cells exhibited a significantly higher ratio of matriptase/HAI-1 mRNA levels compared with HO-8910 cells (0.51 vs. 0.24, ~2.2 fold increase). Compared with HO-8910 cells, the matriptase mRNA level was increased by ~3.6 fold in HO-8910PM cells, whereas the HAI-1 mRNA level was increased by ~1.7 fold. Similar increases in protein expression levels were also observed in HO-8910PM cells compared with HO-8910 cells. Migration and invasiveness were positively correlated with matriptase expression level (r=0.994, P<0.01) and the ratio of matriptase/HAI-1 (r=0.929, P<0.01). Downregulation of matriptase using siRNA resulted in inhibition of the invasive and metastatic abilities of HO-8910PM cells, cell cycle arrest in the G0/G1 phase and increased apoptosis. The present study demonstrated that ovarian cancer cell metastasis and invasion were more dependent on upregulation of matriptase levels than downregulation of HAI-1. Matriptase may be a potential adjuvant therapeutic target for inhibiting ovarian cancer invasion and metastasis.

As a notable feature of malignant tumors, invasion and metastasis are important events in the process of tumor progression. Amiloride, a synthetic inhibitor of urokinase plasminogen activator (uPA), is involved in these events. To evaluate the therapeutic value of amiloride in cervical cancer, HeLa cells were used as in vitro cellular models. The migration and invasion abilities of HeLa cells, in addition to the mRNA expression of matriptase, uPA, uPA receptor and 72 kDa type IV collagenase (MMP-2), were detected using scratch assays, Transwell chamber assays and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results of RT-qPCR demonstrated that the mRNA expression of uPA and MMP-2 in HeLa cells was downregulated significantly in a dose-dependent manner when incubated with various concentrations of amiloride for 24 h. The migration distance of HeLa cells was significantly shorter at 6, 12 and 24 h following incubation with amiloride (P<0.01), and there was a positive correlation between cell migratory ability and cellular uPA protein expression level (r=0.955, P<0.01). The number of HeLa cells that penetrated the Matrigel following incubation for 24 h with different concentrations of amiloride decreased significantly compared with the control group, indicating that cell invasiveness was positively correlated with the protein expression level of uPA in the cells (r=0.993, P<0.01). The present study demonstrated that amiloride was able to specifically inhibit the mRNA expression levels of uPA in HeLa cells, and sequentially downregulate the mRNA expression of downstream MMP-2 in the uPA system, thereby suppressing the migratory and invasive ability of HeLa cells. Therefore, amiloride may be a promising therapeutic target for the treatment of cervical cancer.

Background: Lower serum homocysteine (Hcy) levels are found to correlate with a better chance of clinical pregnancy and better embryo grades in assisted reproductive technology (ART). However, there is little knowledge on the association between Hcy level and unexplained infertility until now. Methods:A total of 388 infertile women undergoing IVF/ICSI treatments were recruited, including 129 women with unexplained causes (case group) and 259 women with known causes (control group), and the case group was further divided into subgroups A (≤8 μmol/L), B (>8 and <15 μmol/L), and C (≥15 μmol/L) based on the serum Hcy level. The associations between serum Hcy level and IVF/ICSI pregnancy outcomes were examined in infertile women with unknown causes.Results: A significantly higher serum Hcy level was measured in the case group than in the control group (P = .008). Subgroup analysis revealed a significant difference in the total number of oocytes retrieved among subgroups A, B, and C (P = .031), and no significant difference was seen among these three groups in terms of age, BMI, E 2 level on the hCG day, number of M-II oocytes, number of fertilized oocytes, or total number of high-quality embryos (P > .05). Spearman correlation analysis revealed a negative correlation between serum Hcy level and total number of oocytes retrieved (r = −.406, P = .019). Univariate and multivariate linear regression analyses revealed that serum Hcy level had no correlations with any IVF/ICSI outcomes. Conclusion:Serum Hcy level has no associations with IVF/ICSI pregnancy outcomes. K E Y W O R D Sassociation study, homocysteine, in vitro fertilization/intracytoplasmic sperm injection, unexplained infertility

This study aimed to induce the differentiation of isolated and purified adipose-derived stromal cells (ADSCs) into myoblasts, which may provide a new strategy for tissue engineering in patients with stress urinary incontinence (SUI). ADSCs, isolated and cultured ex vivo, were identified by flow cytometry and induced to differentiate into myoblasts in the presence of an induction solution consisting of DMEM supplemented with 5-azacytidine (5-aza), 5% FBS, and 5% horse serum. Cellular morphology was observed under an inverted microscope. Ultrastructural changes occurring during the differentiation were observed by transmission electron microscopy and confocal laser scanning microscopy. Cellular immunohistochemical staining was applied to determine the expression of desmin protein in cells with and without induced differentiation. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to detect mRNA and protein expression, respectively, of sarcomeric and desmin smooth muscle proteins. The results showed that ADSCs were mainly of a spindle or polygon shape. Flow cytometry analysis revealed that ADSCs did not express CD34, CD45, and CD106 but high levels of CD44 and CD90, which confirmed that the cultured cells were indeed ADSCs. After induction with a 5-aza-containing solution, morphological changes in ADSCs, including irregular cell size, were observed. Cells gradually changed from long spindles to polygons and star-shaped cells with microvilli on the cell surface. Many organelles were observed and the cytoplasm was found to contain many mitochondria, rough endoplasmic reticulum (rER), and myofilament-like structures. Cell immunohistochemical staining revealed different levels of desmin expression in each phase of the induction process, with the highest expression level found on day 28 of induction. RT-PCR and Western blot results confirmed significantly higher desmin gene expression in induced cells compared with control cells, but no significant difference between the two groups of cells in sarcomeric protein expression. It was concluded that under specific induction setting, ADSCs can be induced to differentiate into myoblasts, providing a potential new option in stem cell transplantation therapy for SUI.

Background To determine the relationship between the levels of stress biomarkers in cord blood and preeclampsia in a hospital-based population of pregnant patients, and to evaluate the resulting effects on pregnancy outcomes. Measures: This prospective case-control study included a total of 282 severe preeclamptic and 534 normal pregnant women. Fetal blood was collected from the umbilical cord at delivery and was utilized for Malonaldehyde (MDA), reactive oxygen species (ROS, superoxide dismutase (SOD) and homocysteine (Hcy) analysis. For a better assessment, we performed a univariate general linear regression model to control for potential confounders and determined the underlying influencing factors for high MDA and ROS. Moreover, a receiver operating characteristic curve analysis was conducted to determine the cutoff values for identifying severe preeclampsia. Further, the severe PE group was divided into the low or high MDA and low or high ROS subgroups according to their cut-off values. Finally, we created logistic regression models to estimate the adjusted odds ratio for each perinatal outcome in high-MDA and -ROS subgroup. Results MDA and ROS levels are higher in women with severe preeclampsia than in normotensive pregnant patients. However, when adjusted for cord blood Hcy levels, the difference is insignificant. Additionally, both MDA (r = 0.359, p < 0.001) and ROS (r = 0.473, p < 0.001) are positively correlated with cord blood Hcy level. The AUCs of MDA and ROS levels were 0.65 (95% CI: 0.60–0.69) and 0.88 (95% CI: 0.86–0.90), respectively, for predicting severe PE. Higher MDA and ROS levels are associated with increased risks of low Apgar score, admission to the NICU, and assisted ventilation for the newborn. Conclusion Increased levels of cord blood oxidative stress markers are significantly associated with negative effects on newborns. High cord blood Hcy levels might be involved in the observed elevated MDA and ROS concentrations in women with severe PE.

<b><i>Objectives:</i></b> The objective of this study was to determine the relationship between the levels of stress biomarkers in cord blood and pre-eclampsia (PE) in a hospital-based population of pregnant patients and evaluate the effects on pregnancy outcomes. <b><i>Design:</i></b> This was an observational, case-control study. <b><i>Participants/Materials, Setting, Methods:</i></b> This case-control study included 282 patients with severe PE and 534 women with normal pregnancy. The umbilical cord was collected at delivery and tested for malonaldehyde (MDA), reactive oxygen species (ROS), superoxide dismutase, and homocysteine (Hcy) analysis. We performed a univariate general linear regression model analysis to control potential confounders and determined the underlying influencing factors for high MDA and ROS. A receiver operating characteristic curve analysis was conducted to determine the cutoff values for identifying severe PE. Further, the severe PE group was divided into the low- or high-MDA and low- or high-ROS subgroups according to the cutoff values. Finally, we created logistic regression models to estimate the adjusted odds ratio for each perinatal outcome in the high-MDA and high-ROS subgroup. <b><i>Results:</i></b> The levels of MDA and ROS levels were higher in women with severe PE than in normotensive pregnant patients. However, when adjusted for cord blood Hcy levels, the difference was insignificant. Additionally, both MDA (<i>r</i> = 0.359, <i>p</i> &#x3c; 0.001) and ROS (<i>r</i> = 0.473, <i>p</i> &#x3c; 0.001) were positively correlated with the cord blood Hcy level. The areas under the curve of MDA and ROS levels were 0.65 (95% confidence interval [CI]: 0.60–0.69) and 0.88 (95% CI: 0.86–0.90), respectively. Higher MDA and ROS levels were associated with increased risks of a low Apgar score, admission to the NICU, and assisted ventilation for the newborn. <b><i>Limitations:</i></b> The study design led to the exclusion of several participants. <b><i>Conclusions:</i></b> Increased levels of oxidative stress markers in the cord blood might be significantly associated with negative effects on newborns. High levels of Hcy in the cord blood might be associated with elevated MDA and ROS concentrations in women with severe PE.

Background: Severe preeclampsia may affect placental development, and high homocysteine (Hcy) levels are linked to intellectual disability. However, the correlation between perinatal Hcy levels and intellectual ability remains unknown in severe preeclampsia-affected offspring. Objective: We aimed to investigate the intellectual ability in offspring born from preeclamptic mothers and examine the role of prenatal Hcy in the prediction of intellectual disability in preschool-aged offspring. Methods: The IQ scores were compared between 101 children born to mothers with severe preeclampsia and 202 offsprings born to normotensive mothers. Maternal Hcy levels within 7 days prior to delivery and postnatal cord blood Hcy were measured. The associations of Hcy with IQ scores were evaluated, and the optimal cutoff values for predicting intellectual disability in the offspring were estimated. Results: The children born to mothers with severe preeclampsia had a greater postnatal cord blood Hcy than those born from normotensive mothers (P < 0.001), and the mothers with severe preeclampsia presented a higher prenatal Hcy (P < 0.001). The children born to mothers with severe preeclampsia had significantly lower IQ scores than those born from normotensive mothers, and a higher Hcy was associated with a lower IQ in preeclampsiaaffected offspring. The prevalence of intellectual disability was 2.86 times higher in severe preeclampsia-affected offspring than in children born from normotensive mothers, and the prevalence of low IQ was greater in children born to mothers with severe preeclampsia than in those from normotensive mothers. ROC curve analysis showed that both maternal and cord blood Hcy were predictors of intellectual disability, and the optimal cutoff for predicting intellectual disability was 17.7 and 9.75 μmol/L for maternal and cord blood Hcy. Conclusion: Perinatal exposure to severe preeclampsia has an adverse effect on postnatal intellectual development, and high maternal and cord blood Hcy may contribute to this association.