FOXM1 transcriptional regulation of RacGAP1 activates the PI3K/AKT signaling pathway to promote the proliferation, migration, and invasion of cervical cancer cells.

BACKGROUND: Cervical cancer (CC) is the most common gynecological tumor disease in women, which occurs at the junction of cervical squamous columnar epithelium. We investigated the effect and mechanism of transcription factor FOXM1 synergizing RacGAP1 in the proliferation, migration, and invasion of CC cells. METHODS: Here, we analyzed the correlation between FOXM1 and RacGAP1 and the clinicopathological features of 68 CC patients. RT-qPCR was used to assess FOXM1 and RacGAP1 mRNA expression in CC tissues and cells. Cell proliferation was assessed by CCK-8 and EDU assays. Transwell assay was applied to test migration and invasion. Cell apoptosis was evaluated utilizing flow cytometry. ChIP and dual-luciferase reporter assays confirmed the interaction of FOXM1 and RacGAP1. Protein levels of FOXM1 and RacGAP1, as well as PI3K/AKT, were analyzed by Western blot. RESULTS: FOXM1 expression was correlated with FIGO stage and histological grade, and RacGAP1 expression was correlated with histological grade. FOXM1 and RacGAP1 levels were increased in CC tissues, and higher expressed in human CC cell lines than that in an immortalized HPV-negative skin keratinocyte line (HaCaT). Depleted RacGAP1 suppressed CC cell proliferation, migration and invasion, and promoted apoptosis. RacGAP1 was a target gene of FOXM1, and FOXM1 positively regulated RacGAP1 expression. FOXM1 had a synergistic effect with RacGAP1 to exert oncogenic function in CC by activating the PI3K/AKT signaling. CONCLUSION: FOXM1 cooperates with RacGAP1 to induce CC cell proliferation, migration and invasion, inhibit apoptosis, and regulate PI3K/AKT signaling to promote CC progression.

Occurrence and Health Risks of Organic Micropollutants in Tap Water in Dalian.

Organic micropollutants (OMPs) in tap water may pose risks to human health. Previous studies on the potential health risks of OMPs in tap water may have underestimated the potential health risks of OMPs due to their limited coverage in target pollutants and incomplete toxicity data. In this study, tap water samples were collected in 37 sampling sites in Dalian, China. More than 1,200 target pollutants were screened by combining screening analysis and target analysis. A total of 93 OMPs were detected, with concentration summation ranging from 157 to 1.7 × 104 ng/L among different sampling sites. A total of 17 OMPs (12 agrochemicals, 3 pharmaceuticals and personal care products, and 2 other compounds) were detected in over 80% of the sampling sites. Especially, imidacloprid, tebuconazole, and atrazine-desethyl were found in all the sampling sites. Computational toxicology models were adopted to predict the missing toxicity threshold values of the identified chemicals. Noncarcinogenic risks were estimated to be negligible among all the sampling sites, while carcinogenic risks at six sites were above 10-6 but below 10-4, indicating non-negligible risks. Griseofulvin contributed the most to the carcinogenic risk. This study offers valuable insights that can guide future initiatives to safeguard tap water safety.

Differences of functionalized graphene materials on inducing chronic aquatic toxicity through the regulation of DNA damage, metabolism and oxidative stress in Daphnia magna.

Graphene can be modified with functional groups when released into the environment. However, very little is known about molecular mechanisms of chronic aquatic toxicity induced by graphene nanomaterials with different surface functional groups. By using RNA sequencing, we investigated the toxic mechanisms of unfunctionalized graphene (u-G), carboxylated graphene (G-COOH), aminated graphene (G-NH2), hydroxylated graphene (G-OH) and thiolated graphene (G-SH) to Daphnia magna during 21-day exposure. We revealed that alteration of ferritin transcription levels in the "mineral absorption" signaling pathway is a molecular initiating event leading to potential of oxidative stress in Daphnia magna by u-G, while toxic effects of four functionalized graphenes are related to several metabolic pathways including the "protein digestion and absorption" pathway and "carbohydrate digestion and absorption" pathway. The transcription and translation related pathways were inhibited by G-NH2 and G-OH, which further affected the functions of proteins and normal life activities. Noticeably, detoxifications of graphene and its surface functional derivatives were promoted by increasing the gene expressions related to chitin and glucose metabolism as well as cuticle structure components. These findings demonstrate important mechanistic insights that can potentially be employed for safety assessment of graphene nanomaterials.

An Efficient UPLC-MS/MS Method Established to Detect Relugolix Concentration in Rat Plasma.

Relugolix, a gonadotropin-releasing hormone (GnRH) receptor antagonist, has been well studied in the treatment of endometriosis symptomatic. It is mainly metabolized by the CYP3A subfamily of P450 enzymes, while minorly metabolized by CYP2C8. Daidzein in different dose groups exhibited a certain induction on the mRNA expression level of CYP3A4 and resulted in the potent induction of CYP3A4. However, it is still unknown whether daidzein and relugolix interact. We developed an effective ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method to study the effect of daidzein on the pharmacokinetics of relugolix in rats after oral administration of 12 mg/kg relugolix in a single or mixed of 50 mg/kg daidzein. The results showed that the method had respectable linearity (r 2 > 0.999) on the scale of 0.7-1000 ng/mL. The intra-day precision was between 3.0% and 8.4% in this assay, and the inter-day was between 4.0% and 11.7%. The intra-day accuracy was from -4.3% to 6.1%, and the inter-day was 2.9% to 12.1%. Another three key indicators, including the stability, the recovery rate of extraction and the new technique's matrix effect, were perfectly in accord with the test verification rule in the biological medium by the United States Food and Drug Administration. Meanwhile, treatment with daidzein led to a decrease in Cmax and AUC0-t of relugolix by about 15.56% and 21.36%, respectively. Although there was no statistical difference in pharmacokinetic parameters, it reflected the induction trend of daidzein on relugolix metabolism for food-drug interaction. It would provide reference and improvement value for subsequent experiments.

Risk assessments of emerging contaminants in various waters and changes of microbial diversity in sediments from Yangtze River chemical contiguous zone, Eastern China.

Over recent decades, increasing chemical contamination has greatly affected aquatic life and human health, even though most contaminants are present at low concentrations. The large-scale chemical industrial parks (CIPs) concentrated in the Yangtze River Delta account for over half of the total in China, and Jiangsu Province occupies one fifth of the Yangtze River Delta. Inevitably, the ecosystems could be affected by these CIPs. In this study, we collected 35 water and 12 sediment samples from the Yangtze River (Taizhou section) surrounding waters adjacent to concentrated CIPs and determined their cumulative chemical levels to be 0.2 to 28.4 µg/L and cumulative detections to be 11 to 39 contaminants with a median of 20 contaminants. 61 out of 153 screened chemicals were detected from at least one sampling site, and 6 contaminants, mostly semi-volatile organic compounds, appeared at all sites. Among these detected chemicals, di-n-octyl phthalate and dibutyl phthalate were at the highest levels. Ecological assessment revealed that 4-chloroaniline, phenol and dibutyl phthalate possibly would induce adverse effects on Yangtze River (Taizhou) ecosystems. Further aided with an evaluation of integrated biomarker response (IBR) index, it was found that site W06 (downstream of Binjiang CIP wastewater inlet) was the location in greatest need of urgent action. As a result, the microbial diversity of sediments in the Yangtze River mainstream was significantly higher than that of tributaries, where CIPs wastewater entered.

Predicting plant cuticle-water partition coefficients for organic pollutants using pp-LFER model.

Predicting plant cuticle-water partition coefficients (Kcw) and understanding the partition mechanisms are crucial to assess environmental fate and risk of organic pollutants. Up to now, experimental Kcw values are determined for only hundreds of compounds because of high experimental cost. For this reason, computational models, which can predict Kcw values based on chemical structures, are promising approaches to evaluate new compounds. In this study, a large dataset consisting of 279 logKcw values for 125 unique compounds were collected and curated. A poly-parameter linear free energy relationship (pp-LFER) model was developed with stepwise multiple linear regression based on this dataset. The resulted pp-LFER model has good predictability and robustness as indicated by determination coefficient (R2adj,tra) of 0.93, bootstrapping coefficient (Q2BOOT) of 0.92, external validation coefficient (Q2ext) of 0.94 and root mean square error of 0.52 log units. Contribution analysis of different interactions indicated that dispersion and hydrophobic interactions have the highest positive contribution (56%) to increase the partition of pollutants onto plant cuticles. In addition, for organic pollutions containing benzene ring (13-31%), double bond (9-17%) or nitrogen-containing heterocycles (9-17%), π/n-electron pairs interactions exhibit obvious positive contributions to logKcw. In conclusion, the proposed pp-LFER model is beneficial for predicting logKcw of potential organic pollutants directly from their molecular structures.

Suspect screening and risk assessment of pollutants in the wastewater from a chemical industry park in China.

Owing to the production and use of chemicals in chemical industry parks (CIPs), these areas are considered to be highly polluted. However, the type of pollutants presents in the wastewater from CIPs and the risk posed to the environment due to the release of these pollutants remains unclear. In this study, suspect screening was combined with traceability analysis to determine the type of pollutants present in wastewaters at 9 chemical enterprises and wastewater treatment plants (WWTPs) in the CIPs. Additionally, the distribution of nine pollutants from the WWTPs' effluent stage and the risk they posed to the surrounding river was examined through target analysis. Upon conducting suspect analysis, the presence of 65 and 64 chemicals in the 9 chemical enterprises' wastewaters and WWTPs, respectively, was tentatively identified. Traceability analysis of the compounds screened in the effluent from the WWTPs determined that 41 substances were identified as characteristic pollutants of the chemical enterprises, indicating that the suspect screening strategy enabled relatively more efficient identification of the characteristic pollutants compared to traditional quantitative analysis. Targeting analysis combined with ecological risk assessment showed that metolachlor, carbendazim, atrazine, diuron, and chlorpyrifos posed relatively higher risks to aquatic organisms in the surrounding river. Therefore, the refined management of the wastewater treatment plant in the CIPs is necessary.

Effects of lomefloxacin on survival, growth and reproduction of Daphnia magna under simulated sunlight radiation.

Lomefloxacin, an antibacterial agent with known photo-induced toxicity in clinical studies, is frequently detected in aquatic environments. Investigating the photo-induced toxicity of lomefloxacin in aquatic organisms is therefore of importance for assessing its ecological risks. In this study, the effects of lomefloxacin on survival, growth and reproduction of Daphnia magna under simulated sunlight radiation (SSR) were investigated, and the mechanism of action was revealed. Results indicated that SSR containing UV radiation increased the acute toxicity of lomefloxacin to Daphnia magna relative to white fluorescent light irradiation. Under SSR, 100 µM lomefloxacin significantly enhanced reactive oxygen species (ROS) generation and lipid peroxidation, and decreased activities of superoxide dismutase and catalase. The biochemical observations and apparent effects on the organism indicate that oxidative stress plays a central role in the acute photo-induced toxicity. Chronic toxicity results showed that SSR significantly affected growth and reproduction of Daphnia magna, whereas lomefloxacin reduced the damage of UV radiation in SSR through light shielding. This study provides insight into the mechanism of photo-induced toxicity and can support the risk assessment of chemicals in the aquatic environment by including the impacts of sunlight irradiation on toxicity.

[High fat diet induced the expression of SREBP-1, TGF-beta1 and alpha-SMA in renal tubular cells and extracellular matrix accumulation in Wistar rats].

OBJECTIVE: To explore the effect of high fat diet on the expression of sterol regulatory element binding protein-1 (SREBP-1), transforming growth factor-beta1 (TGF-beta1) and alpha-smooth muscle actin (alpha-SMA) in renal tubular cells and extracellular matrix accumulation in Wistar rats. METHODS: The Wistar rats were treated with high fat diet for 12 weeks and renal lipid deposit was detected by the method of Oil Red O staining. The immunohistochemistry and Western blot were used to investigate the expression of SREBP-1, TGF-beta1, alpha-SMA and fibronectin (FN) protein. The expression of SREBP-1 mRNA was determined with in situ hybridization. Masson staining was for the detection of extracellular matrix (ECM) accumulation. RESULTS: The weight of rats raised by high fat diet increased, in company with the high serum glucose, serum triglyceride and serum insulin. The Oil Red O staining revealed that the renal proximal tubular epithelial cells showed significant lipid droplet in high fat diet rats. SREBP-1 protein and mRNA were located in the renal tubular cells and the expressions of high fat diet rats were higher than those of normal control rats. They were respectively 1.88 times and 1.85 times than those of normal control group. TGF-beta1 and alpha-SMA protein were also located in renal tubular cells and high fat diet up-regulated the expression of them. ECM accumulation was detected with Masson staining and the result showed that high fat diet treatment increased interstitial ECM product and FN protein was found high expression. CONCLUSION: High fat diet may induce lipid droplet deposit in renal tubular cells by up-regulation of the expression of SREBP-1, which causes ECM accumulation by increasing the expression of TGF-beta1 and alpha-SMA.