Correlations between China's socioeconomic status, disease burdens, and pharmaceuticals and personal care product levels in wastewater.

The presence of pharmaceutical and personal care products (PPCPs) in domestic wastewater can potentially indicate socioeconomic status and disease burdens. However, current knowledge is limited to the correlation between specific pharmaceuticals and diseases. This study aims to explore the associations between socioeconomic status, disease burdens, and PPCP levels in domestic wastewater at a national level. Samples from 171 wastewater influents across China were used to measure PPCPs, and the per capita consumption of PPCPs was calculated. Results showed that the 31 targeted PPCPs were widely present in wastewater with varying occurrence characteristics. The mean consumption levels of different PPCPs varied greatly, ranging from 0.03 to 110723.15 µg/d/capita. While there were no significant regional differences in the overall pattern of PPCP consumption, 22 PPCPs showed regional variations between Northern China and Southern China. PPCPs with similar usage purposes exhibited similar distribution patterns. Disease burden (70.1%) was the main factor affecting most PPCP consumption compared to socioeconomic factors (26.4%). Through correlation analyses, specific types of PPCPs were identified that were highly associated with socioeconomic status and disease burdens, such as hypertension-bezafibrate, brucellosis-quinolones, sulfonamides, hepatitis-triclosan, triclocarban, socioeconomic development-fluoxetine, and people's living standards-gemfibrozil. Despite some uncertainties, this study provides valuable insights into the relationship between PPCPs in domestic wastewater and socioeconomic status and human health.

Effects of point and nonpoint source pollution on urban rivers: From the perspective of pollutant composition and toxicity.

Wastewater discharge is considered to be one of the anthropogenic factors affecting the water quality of urban rivers. The source and composition of wastewater are complex and diverse, and it is difficult to evaluate its effect on water quality and ecological health of receiving waters. Environmental DNA method can determine all species living in waters by examining DNA sequences, reflecting the impact of water quality changes on aquatic systems. In this study, water samples from two urban rivers were collected in dry and wet seasons, and the composition of pollutants was investigated by nontarget screening. Based on the pollutant composition, compound toxicity prediction and concentration addition model were used to predict the toxicity changes of pollutants in the urban rivers. More than 1500 suspect organic pollutants were nontarget screened, and silafluofen was found to be a major toxicity contributor. Environmental DNA analysis was combined with water quality measure and pollutant toxicity prediction to reveal the effects of pollutants from different sources on aquatic ecosystems. Fish diversity was negatively correlated with the mixed toxicity of organic pollutants, suggesting potential ecological risk in these two urban rivers. Our study developed a water quality assessment method based on pollutant composition and toxicity, and the potential risk of nonpoint source pollutants on aquatic ecosystems should not be neglected.

Priority control sequence of 34 typical pollutants in effluents of Chinese wastewater treatment plants.

The identification of the priority control sequence of pollutants in effluents of wastewater treatment plants (WWTPs) has important implications for the management of water quality. This study chose 34 typical pollutants based on their representativeness and detection rates in municipal wastewater. The occurrence frequency and concentration of these pollutants in 168 Chinese WWTP effluents were measured at the national level. The data on in vitro toxicity (67 assays) and in vivo toxicity (216 species) for target pollutants were obtained from the public toxicity database and our experimental data. An environmental health prioritization index (EHPi) method was proposed to integrate the occurrence frequency, concentration, removal rate, and in vitro and in vivo toxicity to determine the priority control sequence of target pollutants. Ethynyl estradiol, 17ß-estradiol, estrone, diclofenac, and atrazine were the top 5 pollutants identified by the EHPi score. Several pollutants with high EHPi scores showed spatial differences. Besides the EHPi method which was from the single pollutant perspective, the combined toxicity of pollutants (300 pairs of binary combinations) was also measured based on in vitro toxicity assays to evaluate the key pollutants from the pollutant-pollutant interacting perspective. The pollutants (such as ofloxacin and acetaminophen) that could have significant synergetic effects with many other pollutants are worthy of prior attention. This study shed new light on the identification of the priority control sequence of pollutants in WWTP effluents. The results provide meaningful data for the effective management and control of wastewater water quality.

Confined water-encapsulated activated carbon for capturing short-chain perfluoroalkyl and polyfluoroalkyl substances from drinking water.

The global ecological crisis of perfluoroalkyl and polyfluoroalkyl substances (PFASs) in drinking water has gradually shifted from long-chain to short-chain PFASs; however, the widespread established PFAS adsorption technology cannot cope with the impact of such hydrophilic pollutants given the inherent defects of solid-liquid mass transfer. Herein, we describe a reagent-free and low-cost strategy to reduce the energy state of short-chain PFASs in hydrophobic nanopores by employing an in situ constructed confined water structure in activated carbon (AC). Through direct (driving force) and indirect (assisted slip) effects, the confined water introduced a dual-drive mode in the confined water-encapsulated activated carbon (CW-AC) and completely eliminated the mass transfer barrier (3.27 to 5.66 kcal/mol), which caused the CW-AC to exhibit the highest adsorption capacity for various short-chain PFASs (C-F number: 3-6) among parent AC and other adsorbents reported. Meanwhile, benefiting from the chain length- and functional group-dependent confined water-binding pattern, the affinity of the CW-AC surpassed the traditional hydrophobicity dominance and shifted toward hydrophilic short-chain PFASs that easily escaped treatment. Importantly, the ability of CW-AC functionality to directly transfer to existing adsorption devices was verified, which could treat 21,000 bed volumes of environment-related high-load (~350 ng/L short-chain PFAS each) real drinking water to below the World Health Organization's standard. Overall, our results provide a green and cost-effective in situ upgrade scheme for existing adsorption devices to address the short-chain PFAS crisis.

Identification and characterization of diverse isomers of per- and polyfluoroalkyl substances in Chinese municipal wastewater.

Linear and branched isomers of per- and polyfluoroalkyl substances (PFASs) are simultaneously present in the environment. However, isomer profiles of PFASs in municipal wastewater treatment plants (WWTPs) are still unknown because of the limitations of standards. Here, influent and effluent samples from 148 municipal WWTPs in China were collected. Ion mobility spectrometry was introduced into high-resolution mass spectrometry-based suspect screening methods to identify the target and suspect PFAS isomers. A total of 38 branched isomers of 14 typical PFASs were identified in wastewater samples. Linear PFASs had higher detection rates (22.3%-100%) than branched isomers (2.0%-98%). Compared to the influents, proportions of branched isomers of most PFASs (except for perfluoropentanoic acid and perfluorohexanoic acid) increased in the effluents. The conventional biological treatment processes (such as anaerobic-anoxic-aerobic and oxidation ditch treatments) had poor removal efficiency for linear PFASs (<21.4%) and branched isomers (<13.4%). No difference on removal efficiency among treatment processes was found. Furthermore, isomer composition in the WWTPs showed obvious differences between East China region and other regions, and the usage of short-chain PFASs (perfluorobutanesulfonic acid and perfluorohexanesulfonic acid) may be a key factor for driving this difference. This study sheds lights on the identification and characterization of PFAS isomers in WWTPs, which would be useful for development of monitoring and control strategies of PFASs.

Perfluorobutanesulfonate exposure induces metabolic disturbances in different regions of mouse gut.

Perfluorobutanesulfonate (PFBS), an alternative to perfluorooctanesulfonate (PFOS), has raised many health concerns. However, PFBS toxicity in the mammalian gut remains unclear. C57BL/6 mice were exposed to 10 µg/L and 500 µg/L PFBS or 500 µg/L PFOS in their water supply for 28 days. PFBS toxicity in the ileum and colon was explored and compared to that of PFOS. Biochemical analysis showed that tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) levels increased in the ileum exposed to 10 µg/L PFBS, whereas no significant changes were observed in those levels in the colon. Catalase (CAT) activity, malondialdehyde (MDA), TNF-α, and IL-1ß levels increased and glutathione (GSH) levels decreased in the ileum of the 500 µg/L-PFBS group, whereas only MDA levels increased in the colon of the 500 µg/L-PFBS group. The results showed that more severe damage occurred in the ileum than in the colon after PFBS exposure, and these align with the 500 µg/L-PFOS group exposure as well. Furthermore, metabolomic analysis revealed glutathione metabolism as a vital factor in inducing PFBS and PFOS toxicities in the ileum. Steroid hormone and amino acid metabolisms were other important factors involved in PFBS and PFOS toxicities, respectively. In the colon, GSH, pyrimidine, and glucose (especially galactose) metabolism was the main contributor to PFBS toxicity, and sulfur amino acid metabolism was the main pathway for PFOS toxicity. This study provides more evidence of the health hazards due to low-dose PFBS exposure in the mammalian gut.

Effects of perfluorobutane sulfonate and perfluorooctane sulfonate on lipid homeostasis in mouse liver.

Perfluorobutane sulfonate (PFBS), an alternative to perfluorooctane sulfonate (PFOS), has been increasingly used in recent years. However, emerging evidence has raised concerns about the potential health risks of PFBS. Here, the toxicityof low-dose PFBS on livers was explored and compared with that of PFOS. Adult C57BL/6 mice were exposed to 10 µg/L, 500 µg/L PFBS, or 500 µg/L PFOS for 28 days through drinking water. At the phenotypic level, no liver damage was observed in the 10 µg/L PFBS group. The cell apoptosis and decrease of CAT activities were observed in the 500 µg/L PFBS group, while accumulation of lipid droplets, increase of CAT activities and TAG levels were found in the 500 µg/L PFOS group. Lipidomics analysis revealed that 138, 238, and 310 lipids were significantly changed in the 10 µg/L, 500 µg/L PFBS and 500 µg/L PFOS groups, respectively. The two PFBS-treated groups induced similar global lipid changes in a dose-dependent manner, which were distinct from PFOS. Overall, PFBS exposure induced an increase in phosphatidylcholines and sphingomyelins, but a decrease in phosphatidylinositol. PFOS exposure caused an increase in triacylglycerols. This study provides more evidence on the health hazards caused by exposure to low-dose PFBS.

Distribution, source and ecological risk of per- and polyfluoroalkyl substances in Chinese municipal wastewater treatment plants.

Municipal wastewater treatment plants (WWTPs) are sinks of per- and polyfluoroalkyl substances (PFASs) generated by human activities and are also sources of PFASs in aquatic environment. This study analyzed distribution, source and ecological risk of 14 PFASs in influent and effluent samples from 148 Chinese municipal WWTPs. Composition and concentrations of PFASs in the influents and effluents had obvious spatial differences. Fluoropolymer processing aids/wrappers and textile treatments/coatings were found to be the dominant sources in WWTP influents, which accounted for 78.34% of all sources. Consumption structure and metal and transportation equipment manufacturing affected the spatial differences of PFASs in WWTPs. Further, mean removal rate of total PFASs in all WWTPs was -5.45%. The conventional treatment processes can not effectively remove PFASs and no significant difference was found among different treatment processes. However, risk quotient values of PFASs in effluents were all below 0.1, indicating low risk or no risk to aquatic organisms. It should be noted that the composition, source and ecological risk of PFASs in east China were different from the other regions, which need more attentions. This study sheds insights into occurrencesof PFASs in municipal WWTPs, which should be helpful for their control strategy development.

The effects of TRAF6 on proliferation, apoptosis and invasion in osteosarcoma are regulated by miR-124.

The present study aimed to verify tumor necrosis factor receptor­associated factor 6 (TRAF6) as the target gene of microRNA-124 (miR-124). In addition, the expression of miR­124 was investigated in osteosarcoma tissues and cells, and its effects on the biological characteristics of osteosarcoma cells were determined, in order to provide an experimental and theoretical basis for the application of TRAF6 in the treatment of osteosarcoma. A fluorescence reporter enzyme system was used to verify TRAF6 as a target gene of miR­124, and western blotting was used to detect the effects of miR­124 on the protein expression levels of TRAF6 in cells. The expression levels of miR­124 were detected in osteosarcoma tissues and an osteosarcoma cell line (MG­63) by quantitative polymerase chain reaction (qPCR). In addition, a total of 48 h post­transfection of MG­63 cells with a miR­124 mimic, qPCR was used to detect the expression levels of miR­124, and the effects of miR­124 on the viability of MG­63 human osteosarcoma cells was determined using the MTT method. The effects of miR­124 on the cell cycle progression and apoptosis of MG­63 cells were analyzed by flow cytometry, whereas the effects of miR­124 on the migration of MG­63 cells was detected using the Transwell invasion chamber analysis method. A TRAF6 recombinant expression plasmid (pcDNA3.1­TRAF6) was also constructed, and MG­63 cells were transfected with the recombinant plasmid and a miR­124 mimic, in order to further validate the biological role of miR­124 via the regulation of TRAF6. The results of the present study indicated that, compared with in the normal control group, the expression levels of miR­124 were significantly increased in MG­63 cells transfected with a miR­124 mimic (P<0.01). In addition, the luciferase reporter gene system demonstrated that, compared with in the control group, relative luciferase activity was significantly reduced in the miR­124 mimic group (P<0.01). The results of MTT analysis indicated that cell viability was also significantly reduced in response to the overexpression of miR­124 in MG­63 cells (P<0.01). Flow cytometric analysis demonstrated that the proportion of cells in S phase and G2/M phase was significantly decreased (P<0.01) in cells overexpressing miR­124, and the number of apoptotic cells was significantly increased (P<0.01). Furthermore, the results of the Transwell invasion assay suggested that the number of invasive cells was significantly decreased following enhanced expression of miR­124 (P<0.01). In MG­63 cells overexpressing miR­124 and TRAF6, the results of MTT, flow cytometric and Transwell assay analyses demonstrated that the overexpression of TRAF6 had the opposite biological effects compared to miR­124 overexpression. In conclusion, the present study indicated that the expression levels of miR­124 were downregulated in human osteosarcoma tissues and cells, and that miR­124 is associated with negative regulation of TRAF6 expression; therefore, the role of TRAF6 in primary osteosarcoma may be regulated by miR­124. Therapeutic strategies that enhance miR­124 expression or inhibit TRAF6 expression may be beneficial for the treatment of patients with osteosarcoma.

MicroRNA-19 contributes to the malignant phenotypes of osteosarcoma in vitro by targeting Pax6.

This study was conducted to detect the expression of miR-19 and Pax6 (Paired box protein 6) in human osteosarcoma cells and the effects on biological characteristics of osteosarcoma cells. Quantitative real-time polymerase chain reaction was used to detect the expression of Pax6 and miR-19 in normal human osteoblasts (hFOB 1.19) and osteosarcoma cell lines (U2OS, Saos-2, and MG-63). Results showed that miR-19 was significantly upregulated in osteosarcoma cell lines compared with that in hFOB 1.19 cells, while the expression of Pax6 messenger RNA was significantly downregulated. Pax6 was defined as the target gene of miR-19 which was validated by luciferase reporter gene analysis. Results indicated that miR-19 had an interaction with Pax6 3'-untranslated region. At the same time, the protein expression of Pax6 was significantly decreased in the MG-63 cells transfected with miR-19 mimic and was notably enhanced in osteosarcoma MG-63 cells transfected with miR-19 inhibitor. These data suggested that Pax6 was a target of miR-19 in osteosarcoma MG-63 cells. The effects of miR-19 on the biological behavior of MG-63 cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and Transwell assay. Results showed that the downregulation of miR-19 inhibited cell viability, reduced the percentage of cells in S phase and the number of cells passing through the Transwell chamber, and increased the number of apoptotic cells. Western blot analysis showed that the inhibition of miR-19 significantly increased the expression of epithelial proteins (E-cadherin and ß-catenin) and decreased the expression of mesenchymal protein (Vimentin), extracellular signal-regulated kinase, and phosphorylated extracellular signal-regulated kinase in MG-63 cells. MiR-19 inhibitor and Pax6 small interfering RNA were simultaneously transfected into MG-63 cells. Results from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and Transwell assay demonstrated that the inhibition of Pax6 expression in MG-63 cells could reverse the cell biological effects induced by the inhibition of miR-19 expression. Based on these findings, it was suggested that miR-19, upregulated in osteosarcoma cells, negatively regulated the expression of Pax6, which can promote the malignant phenotypes of osteosarcoma cells via activation of the extracellular signal-regulated kinase signaling pathways. Therefore, miR-19/Pax6 may offer potential for use as a target for the treatment of osteosarcoma.

[Association of methylenetetrahydrofolate reductase gene polymorphisms with polycystic ovary syndrome].

OBJECTIVE: To assess the association of C677T and A1298C polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene with the susceptibility to polycystic ovary syndrome (PCOS). METHODS: Blood samples of 115 PCOS patients and 58 fertile women (for whom PCOS has been excluded) were collected for DNA extraction. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for determining the C677T and A1298C polymorphisms. A database has been set up with Epidata and a significance test was performed with a statistical analysis system. RESULTS: A significant difference has been found in the allele frequencies of MTHFR gene 677 C and T polymorphisms between the two groups (P<0.01), for which T allele has increased the risk for PCOS by 2.06 times. Heterozygous and homozygous genotypes at position 677 (CT and TT) were more common among PCOS cases than controls, with an OR of 3.91 (95%CI: 1.70-8.97) and 4.39 (95%CI: 1.77-10.89), respectively. There was no statistical difference in genotypic distribution of MTHFR gene A1298C polymorphism (P>0.05). In the PCOS group, there was a significant difference with an OR of 6.40 (95%CI: 1.71-23.95) for an increased risk of insulin resistance in homozygous C677T mutations (TT) compared with the wild genotype (CC, P<0.01). The PCOS group and the control group also differed significantly in their red blood cell folate levels (P<0.01), but not in serum vitamin B12 and homocysteine levels (P>0.05). CONCLUSION: MTHFR gene C677T polymorphism is associated with PCOS, for which CT and TT genotypes can increase the risk of PCOS. The TT genotype can also increase the risk of insulin resistance in PCOS patients. The A1298C polymorphism of the MTHFR gene is not associated with the occurrence of PCOS. The folate level in red blood cells of PCOS patients is lower, for whom folate should be supplemented.