Untargeted metabolomics reveals the mechanism of quercetin enhancing the bioavailability of ticagrelor.

Ticagrelor is a first-line clinical drug for the treatment of acute coronary syndrome, but its oral bioavailability is relatively low. Flavonoids (polyphenol compounds commonly found in plant foods) seriously affect human metabolism and health. This study compared the effects of quercetin, luteolin and catechin on the pharmacokinetic parameters of ticagrelor and found that quercetin can significantly increase the Cmax and area under the curve from time zero to 36 h (AUC0-36 ) of ticagrelor, that is, quercetin can enhance the bioavailability of ticagrelor, but luteolin and catechin cannot. The difference between the ticagrelor group and the combination of quercetin and ticagrelor was analyzed through untargeted metabolomics methods and multivariate data analysis, which identified changes in the levels of seven metabolites (deoxycholic acid, taurocholic acid, glycocholic acid, glycoursodeoxycholic acid, tryptophan, phenylalanine and kynurenine). Based on the changes of these metabolites, we found that the metabolic pathways of phenylalanine, tyrosine and tryptophan and the biosynthetic pathway of bile acids were changed. A metabolomics study revealed that quercetin improves the oral bioavailability of ticagrelor and that this might rely on changing the metabolic pathways of phenylalanine, tyrosine and tryptophan and the biosynthetic pathway of bile acids. The research results at the metabolic level provide us with a strong basis and direction for further exploring the mechanism underlying quercetin's ability to enhance the bioavailability of ticagrelor, and this may be useful for finding new agents that enhance the bioavailability.

Susceptibility of five different sizes of pathogenfree Litopenaeus vannamei to white spot syndrome virus (WSSV) by intramuscular inoculation.

White spot syndrome virus (WWSV) has become one of the most widespread causes of mortality in commercial shrimp farming. In the present study, we used PCR to determine the shrimp infectious dose 50% endpoint (SID50 ml-1) of a Chinese isolate of WSSV in 5 different sizes of pathogen-free Litopenaeus vannamei inoculated intramuscularly. The lethal dose 50% endpoint (LD50 ml-1) was also determined from the percentage of dead shrimp. The LD50 ml-1 for 2, 4, 6, 8, and 10 cm shrimp were 104.68, 105.7, 106.70, 107.75, and 108.81, respectively, and the SID50 ml-1 were 104.68, 105.70, 106.90, 107.75, and 108.94, respectively. There was no significant difference between the LD50 ml-1 and SID50 ml-1 for each shrimp size, which indicated that all infected shrimp died. The lethal and infectious titer decreased about 1 log10 as shrimp size decreased 1 grade. These data clearly indicate that adult shrimp were more susceptible to WSSV than juvenile shrimp. The horizontal comparison showed that the amount of virus in the shrimp organs increased over the experimental period. The vertical comparison showed that virus quantity was lowest in the organs of 10 cm shrimp and highest in 2 cm shrimp, which indicates that the smaller shrimp had higher levels of viral replication. Hence, the optimal size for WSSV challenge in shrimp inoculated intramuscularly was 2 cm. The determination of virus titers in different sizes of shrimp represents a step towards creating strategies to reduce the negative impacts of WSSV in the aquaculture industry.

Involvement of WNT2 in trophoblast cell behavior in preeclampsia development.

This study aimed to determine the WNT2 expression in patients with severe preeclampsia and to explore the function of WNT2 dysregulation on the biological behaviors of trophoblast cells. The WNT2 and ß-catenin expression in the patients with early-onset and late-onset severe preeclampsia and normal controls was determined. Subsequently, WNT2 was overexpressed and knocked down in HTR8 cells and WNT2 signaling pathway in regulating trophoblast cell proliferation, migration, invasion, and apoptosis were evaluated in vitro. The mRNA and protein expression levels of WNT2 and ß-catenin were decreased in patients with preeclampsia, especially early-onset severe preeclampsia. Overexpression of WNT2 promoted trophoblast cell proliferation, migration, and invasion and inhibited apoptosis in vitro, whereas knockdown of WNT2 had opposite effects. The findings of this study reveal that WNT2 and ß-catenin were decreased expressed in patients with preeclampsia. Decreased expression of WNT2 may inhibit trophoblast cell proliferation, migration, and invasion but induced apoptosis. WNT2 may serve as a promising biomarker for early detection of preeclampsia.

Efficient preparation of nitrogen-doped fluorescent carbon dots for highly sensitive detection of metronidazole and live cell imaging.

Herein, nitrogen-doped carbon dots (N-CDs) emitting blue fluorescence were prepared using L-tartaric acid and triethylenetetramine through a simple and quick microwave-assisted method. The synthesized N-CDs displayed excitation-dependent fluorescence behavior, and their maximum excitation and emission wavelengths were 350 and 425 nm, respectively. The obtained N-CDs, which featured excellent fluorescence properties with a high fluorescence quantum yield of 31%, were applied to detect metronidazole (MNZ), which can effectively quench the fluorescence intensity of N-CDs due to the inner filter effect. This phenomenon was used as basis to develop a label-free fluorescent method for rapid MNZ determination, with the limit of detection of 0.22 µM and corresponding linear range of 0.5-22 µM. Hence, we had established a fluorescence method for MNZ detection and applied it to detect MNZ in real samples with satisfactory results. Finally, N-CDs with superior biocompatibility were applied for cell imaging and MNZ detection by the changes in fluorescence intensity.

Post hysteroscopic progesterone hormone therapy in the treatment of endometrial polyps.

OBJECTIVE: To find out the clinical effects of post hysteroscopic progesterone hormone therapy in the treatment of endometrial polyps in terms of clinical outcome and the expression of endometrial Vascular Endothelial Growth Factor (VEGF). METHODS: Ninety-eight patients who were confirmed as endometrial polyp in the hospital from April 2014 and December 2016 were selected and divided into treatment group and a control group using random number table, 49 in each group. Patients in both groups were given hysteroscopic operation. Patients in the treatment group were treated by progesterone hormone drugs after hysteroscopic operation, while patients in the control group were not given progesterone hormone. The changes of menstrual blood volume, menstrual cycle and expression of VEGF were compared between the two groups after treatment, and the recurrence condition, thickness of endometrium and hemoglobin were followed up one year after treatment. RESULTS: The pictorial blood loss assessment chart (PBAC) scores of patients in the two groups had no significant difference before treatment (P>0.05); but the score of the treatment group was much lower than that of the control group. The improvement rate of menstrual cycle of the treatment group was much higher than that of the control group, and the difference had statistical significance (P<0.05). Compared to before treatment, the serum VEGF level of the patients in both groups had a remarkable decline in the 1st, 3rd and 6th month after treatment, and the difference had statistical significance (P<0.05). The difference of the serum VEGF level between the two groups in the 1st and 3rd month after treatment had no statistical significance (P>0.05). The serum VEGF level of the treatment group was notably lower than that of the control group six months after treatment, and the difference had statistical significance (P<0.05). The follow-up results demonstrated that the treatment group had smaller thickness of endometrium and higher level of hemoglobin compared to the control group, and the recurrence rate of the treatment group was lower than that of the control group (P<0.05). CONCLUSION: Post hysteroscopic progesterone hormone therapy has favorable clinical effect in treating endometrial polyps as it can effectively prevent the recurrence of endometrial polyps, relieve the level of hemoglobin and reduce endometrial thickness.

Facile and green synthesis of fluorescent carbon dots with tunable emission for sensors and cells imaging.

Most carbon dots (CDs) conventional fabrication approaches produce single colored fluorescent materials, different methods are required to synthesize distinct carbon dots for specific optical applications. Herein, using one-pot hydrothermal treatment of Syringa obtata Lindl, a facile, low-cost and green assay is achieved in the controllable synthesis of blue and green fluorescent carbon dots. The fluorescent emission of CDs can be well-tuned by adding sodium hydroxide in the precursor solution. Blue fluorescent CDs are applied to Fe3+ sensing with a low detection limit of 0.11 µM of linear range from 0.5 to 80 µM, and then further extended to analysis river water samples. Green fluorescent CDs can be applied to pH detection, which show a remarkable linear enhancement in the green fluorescence emission region when the pH is increased from 1.98 to 8.95. Eventually, the detection of Fe3+ and pH are applied for the living cells fluorescent images in MCF-7 cells are achieved successfully, indicating as-synthesized CDs potential toward diverse application as promising candidate.

Asiatic acid inhibits LPS-induced inflammatory response in endometrial epithelial cells.

Accumulating evidence indicates that asiatic acid, a natural triterpene isolated from Centella asiatica, has anti-inflammatory activity. However, the anti-inflammatory effects of asiatic acid on LPS-stimulated endometrial epithelial cells and the involved molecular pathways have not been completely elucidated. In the present study, we evaluated the effects of asiatic acid on LPS-induced inflammatory response in endometrial epithelial cells. Mouse endometrial epithelial cells were treated with asiatic acid and stimulated with LPS. ELISA was performed to measure the levels of inflammatory cytokines TNF-α, IL-1ß, and PGE2. Western blot analysis was used to test the expression of PPARγ and NF-κB. The results showed that LPS-induced inflammatory mediators TNF-α, IL-1ß, NO, and PGE2 were significantly inhibited by asiatic acid. Furthermore, LPS-induced TLR4 expression and NF-κB activation were concentration-dependently suppressed by asiatic acid. In addition, asiatic acid was found to increase the expression of PPARγ in a concentration-dependently manner. The inhibition of asiatic acid on inflammatory mediators production were prevented by PPARγ inhibitor, GW9662. Taken together, these results showed that asiatic acid exhibited its anti-inflammatory effects in endometrial epithelial cells by activating PPARγ.

Suppressive efficiency of RASSF1A in endometrial carcinoma via inhabiting estrogen receptor alpha expression and ERK pathway activation.

INTRODUCTION: Known as a tumor suppressor, the Ras association domain family 1 isoform A (RASSF1A) is implicated in many human cancers, such as endometrial carcinoma. There is little known about the tumor inhibitive effects of RASSF1A on endometrial carcinoma. The present study was designed to investigate the role of RASSF1A in HEC-1-A cells and to explore its potential mechanisms. MATERIALS AND METHODS: In this study, overexpression of RASSF1A was established by transfection the recombinant adenoviral RASSF1A in HEC-1-A cells. Cells viability was assessed by MTT assay and the apoptosis was analyzed using flow cytometry. Cell migration and invasion were measured in Transwell assay. The levels of ERα and PELP1 protein and extracellular regulated protein kinase (ERK) pathway activation were detected by Western blot. RESULTS: RASSF1A over-expression could significantly inhibit the proliferation, migration and invasion of the HEC-1-A cells in transfection with RASSF1A group compared to that in transfection with control group, also induced apoptosis and suppressed the tumor growth after injection in nude mice. Moreover, overexpression of RASSF1A could inhibit the ERK signal pathway activation and decrease the ERα and PELP1 expression. CONCLUSION: Tumor suppressive efficiency of RASSF1A is exerted through the regulation of ERK pathway activation, ERα and PELP1 expression.

Identification of key genes in endometrioid endometrial adenocarcinoma via TCGA database.

BACKGROUND: Understanding the molecular mechanisms is important in development and therapy of endometrioid endometrial adenocarcinoma. OBJECTIVE: To identify key genes in endometrioid endometrial adenocarcinoma. METHODS: The data of mRNA, miRNA and DNA methylation were downloaded from The Cancer Genome Atlas (TCGA) database and differential analysis was performed. Then, bioinformatic analysis was used to explore the regulatory mechanisms of miRNA and DNA methylation on gene expression. The regulatory network between differentially expressed miRNAs and target genes was established. Finally, the quantitative RT-PCR was applied to validate the bioinformatics results. RESULTS: We obtained biological omics data of 381 patients with endometrioid endometrial adenocarcinoma from TCGA data portal. After data processing, up to 2068 DEGs and 69 differentially expressed miRNAs were identified. Prediction and correlation analysis revealed that 175 DEGs that were not only the target genes but also negatively correlated with the screened differentially expressed miRNAs. After the integrated analysis of differentially methylated CpG islands and DEGs, 16 related genes were obtained. The quantitative RT-PCR results were roughly consistent with the bioinformatics analysis. CONCLUSIONS: The altered DEGs (ZEB1, ZEB2, TIMP2, TCF4, CYP1B1, PITX1, PITX2, ZNF154 and TSPYL5) may be involved in tumor differentiation of endometrioid endometrial adenocarcinoma and could be used as potential therapeutic targets for the disease.

B7-H4 expression promotes tumorigenesis in ovarian cancer.

INTRODUCTION: It has been previously shown that B7-H4, one of the B7 family members that serve as negative regulators of T cell function, has altered expression levels in a variety of cancers, overexpression of B7-H4 promotes cellular transformation. However, there is still lack of adequate evidence to establish a direct connection between B7-H4 expression and malignant transformation. METHODS: Herein, we constructed pE-green fluorescent protein-N1/B7-H4 mammalian expression vector and transfected into B7-H4-negative human ovarian cancer cell line SKOV3. Cellular proliferation, apoptosis, adhesion, motility, and invasion were examined in vitro. Cells injected subcutaneously into severe combined immunodeficient mouse were analyzed for the possible functions of B7-H4 in ovarian tumorigenesis in vivo. RESULTS: Fluorescence microscopy studies confirmed that the B7-H4-green fluorescent protein localizes in the cytoplasm of SKOV3/B7-H4 cells, whereas green fluorescent protein is uniformly distributed throughout the cell. B7-H4 promoted cellular proliferation rate and increased cell adhesion, migration, and invasion. In addition, SKOV3 cells expressing B7-H4 gained growth advantage in the xenograft model in vivo. CONCLUSIONS: These studies demonstrate that B7-H4 directly promotes malignant transformation of ovarian cancer cell line, and provides a potential therapeutic strategy for targeting B7-H4 to inhibit progression of human ovarian cancers.

Altered expression of DACH1 and cyclin D1 in endometrial cancer.

The altered expression of human DACH1, a Drosophila Dachshund homolog, has been associated with tumor progression and metastasis. DACH1 inhibits breast cancer cellular proliferation via cyclin D1. Endometrial cancer is the third most common cancer in women and broad screening for DACH1 expression will further our understanding of this disease. Herein, we screened 126 hysterectomy specimens for DACH1 expression and evaluated the correlation between DACH1 levels and several clinical parameters. Decreased DACH1 expression was significantly correlated with FIGO surgical stages (Stage I-II vs. stage III-IV, p = 0.017), peritoneal cytology (p = 0.044), lymph node positivity (p = 0.035) and histological type (p = 0.007), but not histological grade, depth of myometrial and patient age. Immunostaining was also conducted to examine the expression of cyclin D1, estrogen receptor alpha (ERalpha) and progesterone receptor (PR) in these specimens. Multivariate analysis using the stepwise Cox proportional hazard model showed that FIGO surgical stage, histological grade, lymph node metastasis, and PR expression were correlated with poor survival. Despite the fact that univariate analysis demonstrated that DACH1 positivity is associated with increased 5-year survival in all patients (p = 0.037), decreased expression of DACH1 had no significant value as an independent prognostic factor in predicting survival in endometrial cancers. Our results suggested that loss of DACH1 expression might be involved in endometrial cancer progression.