Prenatal exposure of diabetes and progestin-mediated autistic biomarker in peripheral blood mononuclear cells.

Despite the importance of early diagnosis and intervention, the diagnosis of autism spectrum disorders (ASDs) remains delayed as it is mostly based on clinical symptoms and abnormal behaviours appearing after 2 years of age. Identification of autistic markers remains a top priority in achieving an early and effective ASD diagnosis. We have previously reported that prenatal exposure of hormones or diabetes triggers epigenetic changes and oxidative stress, resulting in gene suppression with autism-like behaviours in offspring. Here, a potential biomarker for ASD diagnosis was established through gene analysis in peripheral blood mononuclear cells (PBMCs). The study from in vivo mouse showed that prenatal hormone exposure or maternal diabetes suppresses mRNA expression of estrogen-related receptor α (ERRα), superoxide dismutase 2 (SOD2), G protein-coupled estrogen receptor (GPER) and retinoic acid-related orphan receptor α (RORA) in the brain as well as oxidative stress and mitochondrial dysfunction, subsequently triggering autism-like behaviour in mouse offspring. Also, similar gene suppression was found in hematopoietic stem cells (HSCs) and PBMC, with inherited epigenetic changes being identified on the related promoters. The human case-control study found that mRNA levels of ERRα, SOD2, GPER and RORA were significantly reduced in PBMC from ASD subjects (n = 132) compared with typically developing (n = 135) group. The receiver operating characteristic curve showed a .869 ± .021 of area under the curve for ASD subjects with 95% confidence interval of .829-.909, together with 1.000 of sensitivity and .856 of specificity. In conclusion, the combined mRNA expression in PBMC based on prenatal factor exposure-mediated gene suppression could be a potential biomarker for ASD diagnosis.

FBXL16 Promotes Endometrial Progesterone Resistance via PP2AB55α /Cyclin D1 Axis in Ishikawa.

Background: F-box proteins are essential components of the E3 ubiquitin ligases which are involved in the regulation of almost all life activities such as cell cycle, proliferation, and apoptosis, which have become an important research and drug target. However, there are few studies on F-box and leucine-rich repeat protein 16 (FBXL16) in endometrial carcinoma. Methods: Clinical samples were collected for determining the correlation between FBXL16 and endometrial carcinoma. Cells were screened and established with Ishikawa cells which proved the fundamental role of FBXL16 in regulating cell proliferation and cell cycle. The MPA-resistant endometrial carcinoma cell line Ishikawa/MPA was established. FBXL16, PP2AB55α , and cyclin D1 were analyzed separately in MPA sensitive and resistant Ishikawa cells in vitro and in vivo. Results: The high expression of FBXL16 was positively correlated with MPA resistance and poor prognosis of endometrial cancer. MPA tolerance of endometrial cancer cells was inhibited by knockdown of FBXL16 in DNA content assessment, CCK-8, and colony formation. It was confirmed that FBXL16 inhibited the activity of substrate PP2AB55α by binding to PP2A, reduced the phosphorylation level at Thr308 site of AKT1, inhibited the expression of GSK-3ß, and thus led to a significant decrease in the phosphorylation level of cyclin D1, which prevented the ubiquitination recognition and degradation of cyclin D1. Conclusion: In our experiments, FBXL16 binds PP2A to promote the dephosphorylation of Thr286 site of cyclin D1 via AKT1/GSK3ß/cyclin D1 pathway, which is required for resisting the ubiquitination degradation and enhances the MPA resistance of Ishikawa.

Curcumenol Targeting YWHAG Inhibits the Pentose Phosphate Pathway and Enhances Antitumor Effects of Cisplatin.

Objective: Cervical cancer is a common cancer in women. The drug resistance of chemotherapeutic agents has always been an urgent problem to be solved in clinics. The purpose of this study was to determine the role of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein gamma polypeptide (YWHAG) in cervical cancer and explore the effect of Curcuma on cervical cancer and its possible mechanism. Methods: YWHAG expression in cervical cancer was confirmed using The Cancer Genome Atlas (TCGA) database. Then, the effects of YWHAG on the proliferation and invasion of HeLa and C33A cervical cancer cells were detected by the cell counting kit-8 (CCK-8) and transwell assay. The relationship between YWHAG and the pentose phosphorylation pathway was further studied. CCK-8, Edu, and quantitative real-time polymerase chain reaction were used to confirm that Curcuma inhibited the sensitivity of YWHAG to cisplatin chemotherapy and to detect the expression of apoptosis-related proteins. Results: YWHAG was highly expressed in cervical cancer and was associated with poor prognosis. The proliferation and invasion abilities of HeLa and C33A cells decreased after YWHAG knockout. The TCGA database of cervical cancer showed a positive correlation between YWHAG and hypoxia-inducible factor-1 subunit alpha (HIF-1α) expression. YWHAG expression increased with HIF-1α overexpression. YWHAG knockdown reduced the protein expression in the pentose phosphorylation pathway. Curcumenol inhibited YWHAG expression. Compared with cisplatin alone, curcumenol combined with cisplatin can reduce cell proliferation and invasion and reduce matrix metalloproteinase (MMP) 2 and MMP9 expression. It can also increase apoptosis, decrease B cell lymphoma 2 (Bcl-2) expression, and increase the expression of Bcl-2 antagonist X, caspase-3, and polyadenosine diphosphate-ribose polymerase. Conclusion: YWHAG can interact with HIF-1α to affect the proliferation and invasion of cervical cancer cells. YWHAG knockout can reduce the expression of pentose phosphorylation pathway-related proteins. Curcumenol can enhance cisplatin to inhibit cancer cell proliferation, migration, and invasion and promote tumor cell apoptosis. The combination of drugs may promote the apoptosis of cervical cancer cells through the YWHAG pathway.

CircZNF124 regulates cell proliferation, leucine uptake, migration and invasion by miR-199b-5p/SLC7A5 pathway in endometrial cancer.

BACKGROUND: Recent studies have revealed that circular RNA participates in endometrial carcinoma (EC) progression. Here we investigated the role of circRNA zinc finger protein 124 (circZNF124) in EC genesis and underlying mechanism. METHODS: The expression levels of circZNF124, microRNA-199b-5p (miR-199b-5p) and solute carrier family 7 member 5 (SLC7A5) were detected by quantitative real-time polymerase chain reaction. The expression of SLC7A5 and other indicated marker proteins was determined by western blot analysis. For functional assay, cell proliferation, leucine uptake and metastasis were investigated by total cell number, cell counting kit-8, cell colony formation, leucine uptake or transwell assay. The interaction between miR-199b-5p and circZNF124 or SLC7A5 was predicted by starbase online database, and identified by mechanism assays. The impact of circZNF124 absence on tumor growth in vivo was revealed by xenograft mouse model assay. Immunohistochemistry assay was implemented to detect the positive expression rate of nuclear proliferation marker (Ki67). RESULTS: CircZNF124 and SLC7A5 expression were significantly increased, while miR-199b-5p was decreased in EC tissues and cells compared with normal endometrial tissues or cells. CircZNF124 expression was closely associated with EC severity and lymph node metastasis. Additionally, circZNF124 depletion repressed cell proliferation, leucine uptake, migration and invasion in both HEC1A and Ishikawa cells. CircZNF124 regulated SLC7A5 expression by binding to miR-199b-5p. MiR-199b-5p inhibitors or SLC7A5 overexpression attenuated circZNF124 silencing-mediated EC malignant progression. Furthermore, SLC7A5 absence inhibited tumor growth in vivo. CONCLUSION: CircZNF124 depletion inhibited EC cell malignancy by miR-199b-5p/SLC7A5 pathway, which demonstrated that circZNF124 had the potential as a therapeutic target for EC.

ERß Accelerates Diabetic Wound Healing by Ameliorating Hyperglycemia-Induced Persistent Oxidative Stress.

Delayed wound healing in diabetic patients is a serious diabetic complication, resulting in major health problems as well as high mortality and disability. The detailed mechanism still needs to be fully understood. In this study, we aim to investigate potential mechanisms and explore an efficient strategy for clinical treatment of diabetic wound healing. Human umbilical endothelial cells were exposed to hyperglycemia for 4 days, then switched to normoglycemia for an additional 4 days. The cells were harvested for the analysis of reactive oxygen species (ROS) generation, gene expression and VEGF signaling pathway. Furthermore, the diabetic wound model was established in rats for the evaluation of wound healing rates under the treatment of either ERß agonist/antagonist or SOD mimetic MnTBAP. Our results show that transient hyperglycemia exposure results in persistent ROS overgeneration after the switch to normoglycemia, along with suppressed expression of ERß, SOD2, and the VEGF signaling pathway. Either ERß expression or activation diminishes ROS generation. In vivo experiments with diabetic rats show that ERß activation or SOD mimetic MnTBAP diminishes ROS generation in tissues and accelerates diabetic wound healing. Transient hyperglycemia exposure induces ROS generation and suppresses ERß expression, subsequently resulting in SOD2 suppression with additional elevated ROS generation. This forms a positive-feed forward loop for ROS generation with persistent oxidative stress. ERß expression or activation breaks this loop and ameliorates this effect, thereby accelerating diabetic wound healing. We conclude that ERß accelerates diabetic wound healing by ameliorating hyperglycemia-induced persistent oxidative stress. This provides a new strategy for clinical treatment of diabetic wound healing based on ERß activation.

PGC1ß Regulates Breast Tumor Growth and Metastasis by SREBP1-Mediated HKDC1 Expression.

Background: Breast cancer is a very common cancer with significant premature mortality in women. In this study, we show that HKDC1 expression in breast cancer cells is increased significantly. We aim to investigate the detailed mechanism for the regulation of HKDC1 expression and its potential contribution to tumorigenesis. Methods: Gene expression was evaluated by real time PCR, western blotting, and immunohistochemistry. The mechanism for PGC1ß/SREBP1-mediated HKDC1 expression was investigated using luciferase reporter assay, chromatin immunoprecipitation, and siRNA techniques. In addition, HKDC1 was overexpressed or knocked down by lentivirus to evaluate the potential effect on in vitro cell proliferation, glucose uptake, mitochondrial function, apoptosis, and reactive oxygen species (ROS) formation. Furthermore, an in vivo xenograft tumor development study was employed to investigate the effect of HKDC1 on tumor growth and mouse survival. Results: HKDC1 is highly expressed in both breast cancer cells and clinical tumor tissues. HKDC1 expression is upregulated and co-activated by PGC1ß through SREBP1 binding motif on the HKDC1 promoter. HKDC1 is located on the mitochondrial membrane and regulates the permeability transition pore opening by binding with VDAC1, subsequently modulating glucose uptake and cell proliferation. Overexpression of HKDC1 increases while knockdown of HKDC1 decreases in vitro breast cancer cell proliferation and in vivo tumor growth, metastasis, and mouse survival. Conclusions: PGC1ß regulates breast cancer tumor growth and metastasis by SREBP1-mediated HKDC1 expression. This provides a novel therapeutic strategy through targeting the PGC1ß/HKDC1 signaling pathway for breast cancer treatment.

Prenatal Progestin Exposure Is Associated With Autism Spectrum Disorders.

We have previously reported that prenatal progestin exposure induces autism-like behavior in offspring through ERß (estrogen receptor ß) suppression in the brain, indicating that progestin may induce autism spectrum disorders (ASD). In this study, we aim to investigate whether prenatal progestin exposure is associated with ASD. A population-based case-control epidemiology study was conducted in Hainan province of China. The ASD children were first screened with the Autism Behavior Checklist (ABC) questionnaire, and then diagnosed by clinical professionals using the ASD diagnosis criteria found in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-V). Eventually, 235 cases were identified as ASD from 37863 children aged 0-6 years old, and 682 matched control subjects with typically developing children were selected for the analysis of potential impact factors on ASD prevalence using multivariate logistic regression. Our data show that the ASD prevalence rate in Hainan was 0.62% with a boy:girl ratio of 5.4:1. Interestingly, we found that the following factors were strongly associated with ASD prevalence: use of progestin to prevent threatened abortion, use of progestin contraceptives at the time of conception, and prenatal consumption of progestin-contaminated seafood during the first trimester of pregnancy. All the above factors were directly or indirectly involved with prenatal progestin exposure. Additionally, we conducted in vivo experiments in rats to further confirm our findings. Either endogenous (progesterone) or synthetic progestin (norethindrone)-treated seafood zebrafish were used to feed pregnant dams, and the subsequent offspring showed autism-like behavior, which further demonstrated that prenatal progestin exposure may induce ASD. We conclude that prenatal progestin exposure may be associated with ASD development.

[Combined Process of DNBF-O3-GAC for Nitrogen and Phosphorus and Metabolite Advanced Removal].

To improve the quality of the tailings water from a wastewater treatment plant (WWTP), a denitrification biofilter (DNBF) with a composite filler composed of a new slow-release organic-carbon source (SOC-F), sponge iron, and activated carbon was tested. Studies were conducted in the combined process of DNBF-O3-GAC to explore the efficiency of the advanced removal of nitrogen, phosphorus, and microbial metabolite by using synthetic effluent made from running water and chemicals. Corresponding comparative studies were conducted by using the secondary effluent from the WWTP. The microbial population structure in the biofilm of the denitrification biofilter was analyzed by adopting MiSeq high-throughput sequencing technologies. The results indicated that the combination process achieved high efficiency removal of nitrogen, phosphorus, and microbial metabolite. The average removal rate of NO3--N in the simulated and actual water period reached 88.87% and 79.99%, respectively; the average removal rate of TP reached 87.67% and 65.51%, respectively; and the average removal rate of UV254 reached 45.51% and 49.23%, respectively. Each processing unit had different functions. The changes in NO3--N, TN, TP, and TFe mainly occurred in the denitrification biofilter, and the removal of UV254 and the change in the three-dimensional fluorescence intensity mainly occurred in the ozone-activated carbon reactor. The cluster analysis at the genus level indicated that the denitrification system had sulfur autotrophic denitrifying bacteria and heterotrophic denitrifying bacteria. Sulfur autotrophic denitrification increased obviously in the actual water period when relatively lack of carbon sources, and the proportion of Thiobacillus increased from 7.44% to 29.62%. The complementary effect of sulfur autotrophic denitrification and heterotrophic denitrification had extended the use of the new slow-release carbon source.

[Preparation and Phosphorus Removal Mechanism of Highly Efficient Phosphorus Adsorbent Mg/Al-LDO].

Aiming at the problem of phosphorus removal in water, Mg/Al-layered double hydroxides (Mg/Al-LDHs) were synthesized via optimized constant pH co-precipitation method, and highly efficient phosphorus adsorbent Mg/Al-layered double oxide(Mg/Al-LDO) was obtained when it was calcined at high temperature. Based on the adsorption characteristics of phosphorus removal, the study combined Zeta potential, X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) to analyze the changes of isoelectric point, crystal structure and functional group before and after adsorption. In addition, Mg/Al-LDO of phosphorus adsorption mechanism was discussed. The results indicated that using the optimized co-precipitation method in the conditions of Mg/Al=2:1, calcination temperature 450℃, and calcination time 2 h, the Mg/Al-LDO adsorption capacity of phosphate was the best, and the maximum adsorption capacity could reach 176.94 mg·g-1, which was basically consistent with the theoretical adsorption capacity of 191.57 mg·g-1, far higher than those of Mg/Al-LDHs and other phosphorus adsorbents. The results showed that the experimental data has the best fitting result with pseudo-second-order kinetics model. The adsorption process was consistent with Langmuir adsorption isotherm model. The results of Zeta potential, XRD and FTIR showed that phosphorus adsorption of Mg/Al-LDO was accomplished co-operatively by electrostatic attraction, anion in layer, ions exchange, and surface co-ordination.

[Feasibility of 3BER-S Process for the Deep Denitrification in Synch with the Removal of PAEs from Reclaimed Water].

In order to investigate the feasibility of deep denitrification and simultaneous removing phthalate esters (PAEs) in the process of reclaimed water treatment uses three-dimensional biofilm-electrode reactor coupled with sulfur autotrophic deep denitrification technology (3BER-S), the technological characteristics and mechanisms were analyzed based on determining the static adsorption capacity of biofilm cultured active carbon fillers in 3BER-S reactor together with the operation results of dynamic denitrification and simultaneous PAEs removing. The results showed that the average adsorption rates of DBP, DEHP were 85.84% and 97.12% in the biofilm cultured active carbon fillers, the equilibrium adsorption capacities were 0.1426 mg x g(-1) and 0.162 mg(-1) and the time spans of reaching adsorption saturation were 120 min and 60 min, respectively; The existence of PAEs had no obvious effect on denitrification, the reactor effluent concentration of TN was in range of 1-2 mg x L(-1) before and after the addition of PAEs, and the average removal rate of TN reached above 94%; 3BER-S denitrification system showed significant ability in removing PAEs, leading to effluent concentrations of DBP and DEHP of no more than 6 microg x L(-1) with removal rates of above 96%; this was due to the synergistic effect of absorption, biodegradation and electrochemistry. After treatment with 3BER-S technology, DBP and DEHP in simulative municipal secondary effluent met the regulated limitation of The Reuse of Urban Recycling Water Quality Standard for Groundwater Recharge (GB/T 19772-2005).

Restoration of rare earth mine areas: organic amendments and phytoremediation.

Overexploitation of rare earth mine has caused serious desertification and various environmental issues, and ecological restoration of a mining area is an important concern in China. In this study, experiments involving dry grass landfilling, chicken manure broadcasting, and plant cultivation were carried out to reclaim a rare earth mine area located in Heping County, Guangdong Province, China. The prime focus was to improve soil quality in terms of nutrients, microbial community, enzyme activity, and physicochemical properties so as to reclaim the land. After 2 years of restoration, an increase of organic matter (OM), available potassium (K), available phosphorus (P) levels, and acid phosphatase (ACP) activity and a reduction of the available nitrogen (N) level and urease (URE) activity in soil were achieved compared to the original mined land. The nutrients and enzyme activities in soil with 5 years of restoration were close to or surpass those in the unexploited land as control. The bulk density, total porosity, water holding capacity, pH, and electrical conductivity (EC) of soil were improved, and the number of cultivable microorganisms and the bacterial diversity in soil were greatly increased with time during ecological restoration, especially for surface soil. Furthermore, the artificial vegetation stably grew at the restored mining sites. The results indicated that organic amendments and phytoremediation could ecologically restore the rare earth mining sites and the mined land could finally be planted as farmland.