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Municipal sewage treatment plants (MSTPs) are environmental pools for antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), which is cause for growing environmental-health concerns. In this study, the effects of different wastewater treatment processes on microbial antibiotic resistance in four MSTPs were investigated. PCR, q-PCR, and molecular cloning integrally indicated that the tetracycline resistance (tet) genes significantly reduced after activated-sludge treatment. Illumina high-throughput sequencing revealed that the broad-spectrum profile of ARGs and mobile element genes (MGEs) were also greatly decreased by one order of magnitude via activated sludge treatment and were closely associated with each other. Correlations between ARGs and bacterial communities showed that potential ARB, such as Acinetobacter, Bacteroides, and Cloaibacterium, were removed by the activated-sludge process. Sedimentation processes cannot significantly affect the bacterial structure, resulting in the relative abundance of ARGs, MGEs, and ARB in second-clarifier effluent water being similar to activated sludge. A comprehensive study of ARGs associated with MGEs and bacterial structure might be technologically guided for activated sludge design and operation in the MSTPs, to purposefully control ARGs carried by pathogenic hosts and mobility.

Salinity is considered as one of the vital factors affecting the profiles of antibiotic resistance genes (ARGs) in soils, whereby its roles in shaping the antibiotic resistome were still poorly understood. Here, metagenomic analysis was conducted to track the ARGs distributions and dissemination in soils during salt accumulation and desalinization processes. Neutral-salt accumulation for 45 and 90 days significantly increased the relative abundances of ARGs and mobile genetic elements (MGEs) carrying antibiotic resistance contigs (ARCs). The ARGs within antibiotic efflux and target protection families primarily carried by Streptomyces, Nocardioides, Rhodanobacter and Monashia were largely enriched by salinity. The ARGs subtypes of the resistance-nodulation-division (RND) family, ATP-binding cassette (ABC) family, rRNA methyltransferase and other efflux were closely associated with MGEs, contributing to the enrichment of ARGs. Moreover, the ARGs subtypes and transposons were genetically linked with the salt-tolerance mechanisms of organic osmolyte transporters and K

As a highly prevalent disease among women worldwide, breast cancer remains in urgent need of further elucidation its molecular mechanisms to improve the patient outcomes. Identifying hub genes involved in the pathogenesis and progression of breast cancer can potentially help to unveil mechanism and also provide novel diagnostic and prognostic markers. In this study, we integrated multiple bioinformatic methods and RNA in situ detection technology to identify and validate hub genes. EZH2 was recognized as a key gene by PPI network analysis. CENPL, ISG20L2, LSM4, MRPL3 were identified as four novel hub genes through the WGCNA analysis and literate search. Among these, many studies on EZH2 in breast cancer have been reported, but no studies are related to the roles of CENPL, ISG20L2, MRPL3 and LSM4 in breast cancer. These four novel hub genes were up-regulated in tumor tissues and associated with cancer progression. The receiver operating characteristic (ROC) analysis and Kaplan-Meier survival analysis indicated that these four hub genes are promising candidate genes that can serve as diagnostic and prognostic biomarkers for breast cancer. Moreover, these four newly identified hub genes as aberrant molecules in the maintenance of breast cancer development, their exact functional mechanisms deserve further in-depth study.

Antibiotics can disturb the gut microbial community and host metabolism. However, their recovery after antibiotics exposure needs to be characterized, and the correlation between gut microbiota and host metabolism remains unclear. In this study, mice were exposed to 0.5, 1.5 and 10 g/L tetracycline hydrochloride (TET) for 2 weeks, then recovered without TET for another 2 weeks. The results showed that 2-week TET exposure changed microbial community and functions in the mouse gut, and increased abundance of antibiotic resistance genes (ARGs), especially in the 10 g/L TET group. After a 2-week recovery, these changes could only be recovered to the control level in the 0.5 g/L TET exposure group, except for ARGs. Besides gut microbiota, TET exposure also changed metabolic profiles in mouse urine. The 2-week recovery significantly reduced changes in metabolic profiles. Some altered metabolites were found to have a very high correlation with gut microbial community and functions, indicating that TET exposure might induce certain changes in urinary metabolic profiles by altering the gut microbiota. The results from this study suggest that the influences of low-level TET exposure are reversible, except for ARGs, which should be paid more attention. During the application of TET, their dosage should be effectively considered and controlled.

BackgroundAs a highly prevalent tumor disease worldwide, Further elucidation of the molecular mechanisms of the occurrence, development and prognosis of breast cancer remain an urgent need. Identifying hub genes involved in these pathogenesis and progression can potentially help to unveil its mechanism and provide novel diagnostic and prognostic markers for breast cancer.MethodsIn this study, we systematically integrated multiple bioinformatic methods, including robust rank aggregation (RRA), functional enrichment analysis, protein-protein interaction (PPI) networks construction and analysis, weighted gene co-expression network analysis (WGCNA), ROC and Kaplan-Meier analyses, DNA methylation analyses and genomic mutation analyses, GSEA and GSVA, based on ten mRNA datasets to identify and investigate novel hub genes involved in breast cancer. In parallel, RNA in situ detection technology was applied to validate those novel hub gene.ResultsEZH2 was recognized as a key gene by PPI network analysis. CENPL, ISG20L2, LSM4 and MRPL3 were identified as four novel hub genes through the WGCNA analysis and literate search. Among those five hub genes, many studies on EZH2 gene in breast cancer have been reported, but no studies are related to the roles of CENPL, ISG20L2, MRPL3 and LSM4 in breast cancer. These novel four hub genes were up-regulated in breast cancer tissues and associated with tumor progression. ROC and Kaplan-Meier indicated these four hub genes all showed good diagnostic performance and prognostic values for breast cancer. The preliminary analysis revealed those novel hub genes are four potentially candidate genes for further exploring the molecular mechanism of breast cancer.ConclusionWe identify four novel hub genes (CENPL, ISG20L2, MRPL3, and LSM4) that are likely playing key roles in the molecular mechanism of occurrence and development of breast cancer. Those hub genes are four potentially candidate genes served as promising candidate diagnostic biomarkers and prognosis predictors for breast cancer, and their exact functional mechanisms in breast cancer deserve further in-depth study.

Background: Further elucidation of the molecular mechanisms of the occurrence, development and prognosis of breast cancer remains an urgent need. Identifying hub genes involved in these pathogenesis and progression can potentially help to unveil these mechanisms and provide novel therapeutic targets for breast cancer. Methods: In this study, we systematically integrated robust rank aggregation (RRA), functional enrichment analysis, protein-protein interaction (PPI) networks construction and analysis, weighted gene co-expression network analysis (WGCNA), DNA methylation analyses and genomic mutation analyses, GSEA and GSVA to identify potential hub genes that are highly associated with breast cancer. Results: We identified a total of 512 robust DEGs that were significantly associated with breast cancer based on RRA analysis and functional enrichment analysis. CENPL, ISG20L2, MRPL3 and LSM4 were identified as four potential hub genes for breast cancer through the WGCNA analysis and literate search. These four hub genes were upregulated in breast cancer tissues and associated with tumor progression. ROC and Kaplan-Meier indicated these four hub genes all showed good diagnostic performance and prognostic values for breast cancer. Methylation analyses and genomic mutation analyses suggested that the abnormal up-regulation of these genes are likelyresulted from hypomethylation and gene mutations. Moreover, GSEA and GSVA for single potential hub genes revealed they were all tightly related to the proliferation of tumor cells. Conclusion: We identify four genes (CENPL, ISG20L2, MRPL3, and LSM4) that are likely playing key roles in the molecular mechanism of occurrence and development of breast cancer. They may become potential therapeutic targets for breast cancer patients with further studies. Keywords: breast cancer, RRA, WGCNA, hub genes

BackgroundColorectal cancer (CRC) is a common malignant solid tumor with an extremely low survival rate after relapse. Previous investigations have shown that autophagy possesses a crucial function in tumors. However, there is no consensus on the value of autophagy-associated genes in predicting the prognosis of CRC patients. This work screens autophagy-related markers and signaling pathways that may participate in the development of CRC, and establishes a prognostic model of CRC based on autophagy-associated genes.MethodsGene transcripts from the TCGA database and autophagy-associated gene data from the GeneCards database were used to obtain expression levels of autophagy-associated genes, followed by Wilcox tests to screen for autophagy-related differentially expressed genes. Then, 11 key autophagy-associated genes were identified through univariate and multivariate Cox proportional hazard regression analysis and used to establish prognostic models. Additionally, immunohistochemical and CRC cell line data were used to evaluate the results of our three autophagy-associated genes EPHB2, NOL3, and SNAI1 in TCGA. Based on the multivariate Cox analysis, risk scores were calculated and used to classify samples into high-risk and low-risk groups. Kaplan-Meier survival analysis, risk profiling, and independent prognosis analysis were carried out. Receiver operating characteristic analysis was performed to estimate the specificity and sensitivity of the prognostic model. Finally, GSEA, GO, and KEGG analysis were performed to identify the relevant signaling pathways.ResultsA total of 301 autophagy-related genes were differentially expressed in CRC. The areas under the 1-year, 3-year, and 5-year receiver operating characteristic curves of the autophagy-based prognostic model for CRC were 0.764, 0.751, and 0.729, respectively. GSEA analysis of the model showed significant enrichment in several tumor-relevant pathways and cellular protective biological processes. The expression of EPHB2, IL-13, MAP2, RPN2, and TRAF5 was correlated with microsatellite instability (MSI), while the expression of IL-13, RPN2, and TRAF5 was related to tumor mutation burden (TMB). GO analysis showed that the 11 target autophagy genes were chiefly enriched in mRNA processing, RNA splicing, and regulation of the mRNA metabolic process. KEGG analysis showed enrichment mainly in spliceosomes. We constructed a prognostic risk assessment model based on 11 autophagy-related genes in CRC.ConclusionA prognostic risk assessment model based on 11 autophagy-associated genes was constructed in CRC. The new model suggests directions and ideas for evaluating prognosis and provides guidance to choose better treatment strategies for CRC.

Wastewater is a major source of biological high-risk pollutants (bHRPs), including pathogenic bacteria, viruses, parasitic protozoa, antibiotic-resistant bacteria, and antibiotic resistance genes. Before wastewater discharge or reclamation, various technologies are being used to remove bHRPs to ensure human health and ecological safety. This chapter is an overview of the main bHRPs' control technologies in wastewater treatment processes that mainly reports the application, action mechanisms, and removal efficiency of conventional disinfection technologies such as chlorination, chlorine dioxide, ozone, ultraviolet, and peracetic acid. For advanced and natural processes, the action mechanisms are also introduced. Furthermore, other technologies including anaerobic and/or aerobic treatment reactor, membrane bioreactor, membrane filter, advanced oxidation processes, constructed wetland, wastewater stabilization pond, nanomaterials, biochar, and bacteriophage are also elucidated. Most of the advanced technologies are interesting but are still in the research state; whereas, conventional disinfection technologies (especially chlorination) are commonly used for the removal of bHRPs. Future work should be focused on new effective technologies including combined technologies to replace conventional disinfection technologies to improve the removal of bHRPs.

Halonitromethanes (HNMs) as one typical class of nitrogenous disinfection byproducts have been widely found in drinking water. In vitro test found HNMs could induce higher cytotoxicity and genotoxicity than trihalomethanes and haloacetic acids. However, data on toxic effect from in vivo experiment is limited. In this study, bromonitromethane (BNM), bromochloronitromethane (BCNM) and trichloronitromethane (TCNM) were chosen as target HNMs, and exposed to mice for 30 d. Hepatic toxicity and serum metabolic profiles were determined to reveal toxic effects and mechanisms of the three HNMs. Results showed the three HNMs significantly decreased relative liver weight, indicating liver is one of the target organs. Further, the three HNMs exposure damaged hepatic antioxidant defense system, and increased oxidative DNA damage. Nuclear magnetic resonance based metabolomics analysis found amino acid metabolism and carbohydrate metabolism were disturbed by HNMs exposure. Some metabolites in these metabolisms are related to oxidative stress and damage. Combined with above results, BNM had the highest toxicity, followed by BCNM and TCNM, indicating bromo-HNMs had higher toxicity than chloro-HNMs. Induction of oxidative stress is one of the toxicity mechanisms of HNMs. This study firstly provides the insight into in vivo toxicity of HNMs and their underlying mechanisms based on metabolomics methods, which is very useful for their health risk assessment in drinking water.

Diabetes is a common metabolic disease, which might influence susceptibility of the kidney to arsenic toxicity. However, relative report is limited. In this study, we compared the influence of inorganic arsenic (iAs) on renal oxidative damage and urinary metabolic profiles of normal and diabetic mice. Results showed that iAs exposure increased renal lipid peroxidation in diabetic mice and oxidative DNA damage in normal mice, meaning different effects of iAs exposure on normal and diabetic individuals. Nuclear magnetic resonance (NMR)-based metabolome analyses found that diabetes significantly changed urinary metabolic profiles of mice. Oxidative stress-related metabolites, such as arginine, glutamine, methionine, and β-hydroxybutyrate, were found to be changed in diabetic mice. The iAs exposure altered amino acid metabolism, lipid metabolism, carbohydrate metabolism, and energy metabolism in normal and diabetic mice, but had higher influence on metabolic profiles of diabetic mice than normal mice, especially for oxidative stress-related metabolites and metabolisms. Above results indicate that diabetes increased susceptibility to iAs exposure. This study provides basic information on differential toxicity of iAs on renal toxicity and urinary metabolic profiles in normal and diabetic mice and suggests that diabetic individuals should be considered as susceptible population in toxicity assessment of arsenic.

The response mechanism of wastewater treatment biofilms to salt stress has not yet been fully established. The aim of this study was to reveal the comprehensive biological effects of salinity on biofilm microbial community and metabonomic characteristics. The study assessed performance at a range of sodium chloride (NaCl) concentrations of 0.6, 14 and 20 g/L. Biofilm coverage rate decreased significantly with increasing NaCl concentrations. High NaCl concentrations resulted in more compact and smoother biofilm morphologies. NaCl concentrations affected bacterial community variation at the class and genus level, with Gammaproteobacteria being the most dominant Proteobacteria, exhibiting NaCl tolerance at concentrations ranging from 0 to 20 g/L. Also, NaCl sensitive or tolerant species were identified, such as Pseudomonas and Planococcus, respectively. Dominant metabolites in wastewater treatment biofilms belonging to nucleotide, lipid, vitamin, amino acid and carbohydrate metabolism pathways decreased with increasing NaCl concentrations. High concentrations of NaCl regulated cell motility, transcription and membrane transport functions. In particular, the activity of ABC transporters were up-regulated at NaCl concentrations of 0.6 g/L and down-regulated at higher salinity concentrations. In addition, transcription machinery were inhibited under the stress of 14 g/L NaCl. These findings further our understanding of the short-term adaption mechanisms of wastewater treatment biofilms to high NaCl concentration environments.

Microcystin-LR (MC-LR) can induce hepatic tissue damages and molecular toxicities, but its effects on lipid metabolism remain unknown. This study investigated the effects of MC-LR exposure on mice lipid metabolism and uncovered the underlying mechanism through metabonomic, transcriptomic and metagenomic analyses after administration of mice with MC-LR by gavage for 28 d. Increased liver weight and abdominal fat weight, and evident hepatic lipid vacuoles accumulation were observed in the mice fed with 0.2mg/kg/d MC-LR. Serum nuclear magnetic resonance analysis showed that MC-LR treatment altered the levels of serum metabolites including triglyceride, unsaturated fatty acid (UFA) and very low density lipoprotein. Digital Gene Expression technology was used to reveal differential expression of hepatic transcriptomes, demonstrating that MC-LR treatment disturbed hepatic UFA biosynthesis and activated peroxisome proliferator-activated receptor (PPAR) signaling pathways via Pparγ, Fabp1 and Fabp2 over-expression. Metagenomic analyses of gut microbiota revealed that MC-LR exposure also increased abundant ratio of Firmicutes vs. Bacteroidetes in gut and altered biosynthetic pathways of various microbial metabolic and pro-inflammatory molecules. In conclusion, oral MC-LR exposure can induce hepatic lipid metabolism disorder mediated by UFA biosynthesis and PPAR activation, and gut microbial community shift may play an important role in the metabolic disturbance.

Graphene is a promising candidate as an antibacterial material owning to its bacterial toxicity. However, little information on influence of graphene on gut microbiota is available. In this study, mice were exposed to graphene for 4 weeks, and high-throughput sequencing was applied to characterize the changes in microbial community and antibiotic resistance genes (ARGs) in mouse gut. The results showed that graphene exposure increased biodiversity of gut microbiota, and changed their community. The 1 μg/d graphene exposure had higher influences on the gut microbiota than 10 μg/d and 100 μg/d graphene exposures, which might be due to higher aggregation of high-level graphene. The influence of graphene on gut microbiota might attribute to that graphene could induce oxidative stress and damage of cell membrane integrity. The results were verified by the increase of ratio of Gram-negative bacteria. Outer membrane of Gram-negative bacteria could reduce the membrane damage induced by graphene and make them more tolerance to graphene. Further, we found that graphene exposure significantly increased the abundance and types of ARGs, indicating a potential health risk of graphene. This study firstly provides new insight to the health effects of graphene on gut microbiota.

Halonitromethanes (HNMs) are a class of halogenated nitrogenous disinfection by-products (N-DBPs) in drinking water which possess health concerns due to their potentially higher toxicity than regulated disinfection by-products (DBPs). A cold on-column (COC) injection in track-oven mode coupled with a gas chromatography-mass spectrometry (GC-MS) system for the analysis of HNMs has been developed. Comparative experiments showed that COC had an advantage over the conventional split/splitless injection in minimizing thermal degradation of HNMs, especially dibromochloro- and tribromo-nitromethanes in water. Both debromo- and denitro-products of HNMs were observed in the splitless injection mode at 117 °C and 170 °C. Liquid–liquid extraction (LLE) and solid phase extraction (SPE) procedures were compared for sample pretreatment. LLE showed good recoveries of 73–91% for all nine HNMs. In comparison, SPE provided a similar recovery range for four commonly detected HNMs in drinking water: dichloronitromethane, trichloronitromethane, bromochloronitromethane and dibromonitromethane in drinking water, while the recoveries of the other HNMs were below 50%. This indicated that the SPE-COC-GC-MS method can be a good alternative to LLE-COC-GC-MS for the identification and quantification of the four HNMs commonly present in tap water due to the simplicity of the SPE pretreatment technique.

Iodinated disinfection byproducts (DBPs) are widely present in disinfected drinking waters and wastewater effluents, and they have drawn increasing concern owing to their high toxicity. To date, the reported iodinated DBPs mainly include aliphatic and aromatic ones, and iodinated trihalomethanes (THMs) and haloacetic acids (HAAs) are the most commonly detected aliphatic iodinated DBPs in disinfected waters. In this study, the formation of iodinated THMs and HAAs from aromatic iodinated DBPs during chloramination was investigated. The decomposition kinetics of the aromatic iodinated DBPs and the formation of iodinated THMs and HAAs were studied, the formation pathways of iodinated THMs and HAAs from the aromatic iodinated DBPs were explored, the factors affecting the formation were examined, and the toxicity change was evaluated. The results revealed that four aromatic iodinated DBPs (2,4,6-triiodophenol, 3,5-diiodo-4-hydroxybenzaldehyde, 3,5-diiodosalicylic acid, and 2,6-diiodo-4-nitrophenol) all underwent transformation to form triiodomethane (TIM), monoiodoacetic acid (MIAA), and diiodoacetic acid (DIAA) during chloramination. The decomposition of the aromatic iodinated DBPs all followed a pseudo-first-order decay during chloramination, and the rank order of the decomposition rate constants was as follows: 2,4,6-triiodophenol > 3,5-diiodo-4-hydroxybenzaldehyde ≥ 3,5-diiodosalicylic acid > 2,6-diiodo-4-nitrophenol. Several polar iodinated intermediates were detected and identified (e.g., 2,6-diiodo-1,4-benzoquinone and iodobutenedioic acid) during chloramination of 2,4,6-triiodophenol, based on which the formation pathways of TIM, MIAA, and DIAA from 2,4,6-triiodophenol during chloramination were proposed and further validated. The results also revealed that monochloramine dose, pH, temperature, and short free chlorine contact time all affected the formation of TIM, MIAA, and DIAA from 2,4,6-triiodophenol during chloramination. The cytotoxicity order of the eight iodinated DBPs was MIAA > 2,6-diiodo-4-nitrophenol > 2,4,6-triiodophenol > 2,6-diiodo-1,4-benzoquinone > DIAA ≥ 3,5-diiodosalicylic acid >3,5-diiodo-4-hydroxybenzaldehyde > TIM. The toxicity of the chloraminated 2,4,6-triiiodophenol sample first decreased and then increased over time due to the transformation.

Halonitromethanes (HNMs) as one typical class of nitrogenous disinfection byproducts in drinking water and wastewater are receiving attentions due to their high toxicity. This study applied a simulator of the human gastrointestinal tract to determine the dehalogenation processes of trichloronitromethane, bromonitromethane and bromochloronitromethane for the first time. Influence of digestion process of HNMs on gut microbiota and hepatotoxicity was further analyzed. Results showed that the three HNMs were rapidly and completely dehalogenated in the gastrointestinal tract, especially in the stomach (2 h retention Time) and small intestine (4 h retention Time). Mucin, cysteine, pancreatin and bile salts in the digestive juice played major roles in the dehalogenation process. HNMs and their dehalogenation products in the resulting fluids of stomach induced the highest toxicity followed by those in intestine and colon, exhibiting dose-dependent effects. Although most HNMs were degraded in the stomach and small intestine, residual HNMs entered into colon changed the microbial community. Abundance of several genera, such as Bacteroides, Lachnospiraceae_unassigned and Lactobacillus had high correlation with exposure concentration of HNMs. This study sheds new light on dehalogenation and toxic processes of HNMs by oral exposure, which provides basic data for their human health risk assessment.

Antibiotics have been widely used for disease prevention and treatment of the human and animals, and for growth promotion in animal husbandry. Antibiotics can disturb the intestinal microbial community, which play a fundamental role in animals' health. Misuse or overuse of antibiotics can result in increase and spread of microbial antibiotic resistance, threatening human health and ecological safety. In this study, we used Illumina Hiseq sequencing, (1)H nuclear magnetic resonance spectroscopy and metagenomics approaches to investigate intestinal microbial community shift and antibiotic resistance alteration of the mice drinking the water containing tetracycline hydrochloride (TET). Two-week TET administration caused reduction of gut microbial diversity (from 194 to 89 genera), increase in Firmicutes abundance (from 24.9 to 39.8%) and decrease in Bacteroidetes abundance (from 69.8 to 51.2%). Metagenomic analysis showed that TET treatment affected the intestinal microbial functions of carbohydrate, ribosomal, cell wall/membrane/envelope and signal transduction, which is evidenced by the alteration in the metabolites of mouse serum. Meanwhile, in the mouse intestinal microbiota, TET treatment enhanced the abundance of antibiotic resistance genes (ARGs) (from 307.3 to 1492.7 ppm), plasmids (from 425.4 to 3235.1 ppm) and integrons (from 0.8 to 179.6 ppm) in mouse gut. Our results indicated that TET administration can disturb gut microbial community and physiological metabolism of mice, and increase the opportunity of ARGs and mobile genetic elements entering into the environment with feces discharge.

Aromatic iodinated disinfection byproducts (DBPs) are a newly identified category of highly toxic DBPs. Among the identified aromatic iodinated DBPs, 2,4,6-triiodophenol and 2,6-diiodo-4-nitrophenol have shown relatively widespread occurrence and high toxicity. In this study, we found that 4-iodophenol underwent transformation to form 2,4,6-triiodophenol and 2,6-diiodo-4-nitrophenol in the presence of monochloramine. The transformation pathways were investigated, the decomposition kinetics of 4-iodophenol and the formation of 2,4,6-triiodophenol and 2,6-diiodo-4-nitrophenol were studied, the factors affecting the transformation were examined, the toxicity change during the transformation was evaluated, and the occurrence of the proposed transformation pathways during chloramination of source water was verified. The results revealed that 2,4,6-triiodophenol and 2,6-diiodo-4-nitrophenol, which could account for 71.0% of iodine in the transformed 4-iodophenol, were important iodinated transformation products of 4-iodophenol in the presence of monochloramine. The transformation pathways of 4-iodophenol in the presence of monochloramine were proposed and verified. The decomposition of 4-iodophenol in the presence of monochloramine followed a pseudo-second-order decay. Various factors including monochloramine dose, pH, temperature, nitrite concentration, and free chlorine contact time (before chloramination) affected the transformation. The cytotoxicity of the chloraminated 4-iodophenol samples increased continuously with contact time. The proposed transformation pathways occurred during chloramination of source water.

This study investigated health risk reduction in a drinking water treatment plant of Nanjing City (China) based on chemical detection of 22 semi-volatile organic compounds (SVOCs) and 24 metallic elements in source water and drinking water during 2009–2011. Chemical analysis showed that 15 SVOCs and 9 metals were present in the water. Health risk assessment revealed that hazard quotient of each pollutant and hazard index (HI) of all the detectable pollutants were below 1.00, indicating that the chemicals posed negligible non-carcinogenic risk to local residents. Benzo(a)pyrene may induce carcinogenic risk since its risk index via both oral and dermal exposure exceeded the safety level (1.00E−6), but other SVOCs induced no carcinogenic risk. Total HI of the SVOCs was 1.08E−3 for the source water and 1.56E−3 for the drinking water, suggesting that the used conventional treatment processes (coagulation/sedimentation, sand filtration and chlorine disinfection) cannot effectively reduce the non-carcinogenic risk. The source water had higher carcinogenic risk than the drinking water, but risk index of the drinking water still exceeded 1.00E−6. This study might serve as a basis for health risk assessment of drinking water and also as a benchmark for the authorities to reduce health risk arising from trace-level hazardous pollutants.

DNA microarray was used to analyze hepatic transcriptional profile of male mice (Mus musculus) after the mice were fed with Yangtze River (China) source water (NJS) and tap water (NJT) for 90 days. Chemical analysis demonstrated that NJS and NJT contained various trace-level pollutants including polycyclic aromatic hydrocarbons, phthalate ethers and inorganic contaminants. DNA microarray revealed occurrence of 5,042 differentially expressed genes (DEGs) in the mice fed with NJS and 828 DEGs in the mice fed with NJT, indicating that NJS posed greater influence on liver transcriptome. Annotation against Kyoto Encyclopedia of Genes and Genomes pathway database showed that the DEGs in NJS group were mostly involved in lipid metabolism (51 DEGs), followed by neurodegenerative diseases (47 DEGs), energy metabolism (41 DEGs) and endocrine system (38 DEGs). NJT exposure was found to affect lipid metabolism (14 DEGs), xenobiotics biodegradation and metabolism (6 DEGs), and cofactors and vitamins metabolism (5 DEGs). Annotation against Gene Ontology database confirmed that lipid metabolism among the altered pathways was most susceptible to both NJS and NJT exposure. The DEGs were involved in 6 lipid metabolic pathways including fatty acid metabolism, glycerophospholipid metabolism, unsaturated fatty acid biosynthesis, steroid biosynthesis, primary bile acid biosynthesis and steroid hormone biosynthesis. Although both NJS and NJT might cause no obvious liver tissue damages, the lipid metabolic disturbance induced by trace-level pollutants still deserves public health concern.

Expression of heparanase is associated with invasion, metastasis and angiogenesis of a variety of human cancers. However, the roles of heparanase in cervical cancer are not clear. The aim of this study is to determine whether up-regulation of heparanase expression can promote growth of cervical cancer in vitro and in vivo. Heparanase protein expression was analyzed in cervical cancer patients using immunohistochemistry. In addition, expression of heparanase was also examined in cervical cancer cell lines. A series of in vivo and in vitro assays was performed to elucidate the role of heparanase in tumor growth of cervical cancer. Positive rate of heparanase was 63.3 % (38/60) in cervical cancer patients by immunohistochemistry, and it was significantly correlated with tumor size and clinical stage (P < 0.05). Overexpression of heparanase inhibited apoptosis of cervical cancer cells. Moreover, ectopic overexpression of heparanase in cervical cancer cells promoted proliferation of cervical cancer cells in vitro and tumor growth in vivo. These results suggest that heparanase participates in tumor growth of cervical cancer by influencing the proliferation and apoptosis of cervical cancer cells, and heparanase could be used as an effective therapeutic target for cervical cancer.

Apoptosis in cultured rat hippocampal neurons was induced using the nitric oxide donor 3-morpholinosydnonimine, and cells were treated with the chloride channel blocker, 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid. Results showed that the survival rate of neurons was significantly increased after treatment with 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid, and the rate of apoptosis decreased. In addition, the expression of the apoptosis-related proteins poly(adenosine diphosphate-ribose)polymerase-1 and apoptosis-inducing factor were significantly reduced. Our experimental findings indicate that the chloride channel blocker 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid can antagonize apoptotic cell death of hippocampal neurons by inhibiting the expression of the apoptosis-related proteins poly(adenosine diphosphate-ribose)polymerase-1 and apoptosis-inducing factor.

Foxp3 was identified as a key protein in mediating inhibitory functions of regulatory T cell (Treg). Foxp3 was thought to express only in the T cell lineage until recently when some researches reported that Foxp3 was also expressed by cancer cells. In this study, we describe for the first time the expression of Foxp3 in cervical cancer. Progression from cervical intraepithelial neoplasia (CIN) to cervical cancer is a multistep process initiated by persistent infection with high-risk human papillomavirus (HPV). P16(INK4a) is a crucial marker of HPV integration into host cells. In the present study, expressions of Foxp3 and P16(INK4a) in CIN and cervical cancer were detected by immunohistochemistry. Our results found expression level of Foxp3 was increased during the progression of cervical neoplasia. Moreover, up-regulation of Foxp3 appeared to be correlated with the expression of P16(INK4a). Examination of the role of Foxp3 in differentiation by double immunostaining for cytokeratin 10 (CK10) showed significant association between Foxp3 expression and differentiation (Foxp3 vs CK10). Furthermore, positive expression of Foxp3 was correlated with tumor size. These data suggest that Foxp3 may play an important role in differentiation and growth of cervical cancer cells. Our findings provide new insights regarding the role of Foxp3 in differentiation and its association with HPV infection during the development of cervical cancer.

Minocylcine, a tetracycline derivate, has been shown to cross the blood-brain barrier and enter the central nervous system. In this study, cerebral ischemia-reperfusion injury models were established using the suture method, and minocycline was immediately injected intraperitoneally after cerebral ischemia-reperfusion (22.5 mg/kg, initially 45 mg/kg) at a 12-hour interval. Results showed that after minocycline treatment, the volume of cerebral infarction was significantly reduced, the number of surviving cell in the hippocampal CA1 region increased, the number of apoptotic cells decreased, the expression of caspase-3 and poly(adenosine diphosphate-ribose) polymerase-1 protein was down-regulated, and the escape latency in the water maze test was significantly shortened compared with the ischemia-reperfusion group. Our experimental findings indicate that minocycline can protect against neuronal injury induced by focal ischemia-reperfusion, which may be mediated by the inhibition of caspase-3 and poly(adenosine diphosphate-ribose) polymerase-1 protein expression.

Protein 14-3-3γ is an important member of the 14-3-3 family that play important roles in the regulation of various cellular processes. The aim of the study is to investigate the association between 14-3-3γ expression and the clinicopathological features of patients with breast cancer.The expression of 14-3-3γ was detected by Western blot in both foci of breast cancer and adjacent non-cancerous tissues. In addition, 14-3-3γ expression was analyzed by immunohistochemistry in 60 clinicopathologically characterized breast cancer cases. The association of 14-3-3γ expression with survival of the patients were analyzed.The expression level of 14-3-3γ protein in breast cancer were significantly higher than that in non-cancerous mammary gland tissues. Moreover, high expression of 14-3-3γ correlated with tumor size and tumor grade (all P0.05). Patients with high 14-3-3γ expression had worse overall survival rate than that with low expression (P0.05). Furthermore, multivariate analysis showed that 14-3-3γ expression was an independent predictor of overall survival (HR, 0.196; 95%CI, 0.043-0.892; P = 0.035).Our data suggest for the first time that the increased expression of 14-3-3γ in breast cancer is associated significantly with tumor progression and poor prognosis. 14-3-3γ may be a novel and potential prognostic marker for breast cancer.

Objective Annexins are a family of intracellular proteins that bind membrane phospholipids in a Ca2 + concentration-dependent manner. Several annexins play important roles during tumor progression. However, little is known about the clinical implications and biological functions of Annexin A3 in breast cancer. Methods Using immunohistochemistry, we analyzed 60 breast cancers for the levels of annexin A3 and investigated the correlation of its expression change with patient's survival via Kaplan–Meier survival analysis. Furthermore, via knockdown of Annexin A3 expression in breast cancer cells with special siRNA, the role of Annexin A3 in the proliferation and apoptosis of breast cancer cells was examined. Results Annexin A3 was expressed at higher level in breast cancer than that in normal breast tissue. The expression of Annexin A3 in human breast carcinoma closely correlated with tumor size and axillary lymph node metastasis. Kaplan–Meier survival analysis revealed a significant inverse correlation between strong Annexin A3 expression and overall patient survival. Moreover, Annexin A3 overexpression was inversely associated with Bax staining and the apoptosis index. Annexin A3 small interfering RNA in MCF-7 and MDA-MB-435 could inhibit cell proliferation, decrease Bcl-2 mRNA and protein expression, and increase Bax mRNA and protein expression. Conclusion Our findings indicated that Annexin A3 might be a novel and potential prognostic marker for patients with breast cancer and be involved in regulating apoptosis by affecting Bcl-2/Bax balance.

OBJECTIVE Through determination of the dynamic change of the active component in different parts of Prunella vulgaris at different growth stages, to find the optimal harvest time. METHOD Total flavonoids content was determined by using the spectrophotometric method, and the content of ursolic acid and oleanolic acid was determined by HPLC. The contents of ash and extract were determined according to the methods in Chinese Pharmacopeia (2005 edition). RESULT There existed the active components in all parts of P. vulgaris, but the active component contents in different parts of P. vulgaris of at different growth stages, changed very obviously. CONCLUSION In Yangtze-Huaihai region, the optimal harvest time of Prunella spike best harvest is at the end of June, and Prunellastem at the end of May. All parts of P. vulgaris have medicinal value.