[The advances on the formation and toxic effects of haloacetaldehydes disinfection by-products in drinking water].

Haloacetaldehydes (HALs), as emerging disinfection by-products in drinking water, are the third largest group by weight of identified disinfection by-products (DBPs) in drinking water. The formation of HALs is associated with the level of natural organic matter and halide in the source water, the treatment process of drinking water and the type of disinfectant. Recent studies have shown that HALs are more cytotoxic and genotoxic than regulated trihalomethanes and halo-acetic acids in drinking water. Currently, only a few countries and regions have set limit values for trichloroacetaldehyde with high detection rate in drinking water. However, there is growing evidence that unregulated HALs have a higher potential risk to human health compared to regulated HALs. This paper reviews the current research progress on the formation and transformation, cytotoxicity and genotoxicity of HALs in drinking water, and looks forward to the problems that should be paid attention in the future toxicological research of HALs in order to support the development of scientific drinking water standards.

[Impact of hypoxia-reoxygenation environment on autophagy level of osteoblasts].

Objective: To investigate the impact of hypoxia-reoxygenation environment on the level of autophagy in osteoblasts. Methods: Osteoblasts were purified from the skulls of newborn SD rats within 24-48 hours by tissue block adherence culture and differential centrifugation. The osteoblasts were identified by alizarin red staining and alkaline phosphatase staining. The third generation osteoblasts were cultured in normal state and randomly divided into four groups: group A was cultured under normal condition for 36 hours; group B was cultured under normal condition for 18 hours, then under hypoxia for 18 hours; group C was cultured under hypoxia for 36 hours; group D was cultured under hypoxia for 18 hours, and then under normal condition for 18 hours. The ability to form calcium nodules of osteoblasts in the four groups was observed after culture. The proliferation activity of osteoblasts was detected by CCK-8 assay. The expressions of autophagy specified gene Beclin 1, microtubule-associated protein light chain 3(LC3) and collagen Ⅰ(COL-Ⅰ), bone morphogenetic protein 2 (BMP-2) genes were detected by real time polymerase chain reaction (RT-PCR), and the protein expressions of Beclin 1, LC3-Ⅰ,LC3-Ⅱ and P62 were detected by immunoblotting. Results: Alizarin red staining showed that osteoblasts in group A had the strongest calcification ability, and calcification ability of osteoblasts in group B,C and D lowered gradually, and it was lowest in group D. The proliferative activity under the CCK-8 detection in group A, B, C and D was 98%±8%, 90%±8%,82%±9%,76%±8%, respectively (F=35.764, P=0.000). The mRNA expression of Beclin 1, LC3-Ⅱthe 4 groups increased gradurally (group D> group C> group B> group A)(F=38.327, 16.583, both P<0.05); and the mRNA expression of COL-Ⅰ, BMP-2 decreased gradually in the 4 groups (group A> group B> group C> group D) (F=20.387, 12.426, both P<0.05). The protein expression of Beclin 1,LC3-Ⅱ/LC3-Ⅰ increased gradually in the groups (group D>group C>group B>group A) (F=26.843, 28.576, bothP<0.05), and the expression of P62 protein decreased gradually (F=18.946, P=0.011). Conclusions: Hypoxia-reoxygenation environment can reduce the proliferation activity of osteoblasts and up-regulate the expression of autophagy-related genes in osteoblasts. Anoxic reoxygenation environment promotes the increasing of autophagy levels in osteoblasts.

[Research progress on exposure levels and toxic pathways of typical persistent organic pollutants in foods].

Dioxins, polybrominated diphenyl ethers, and benzo(a)pyrene are common organic pollutants in food. They have been of concern to academics and government administrations due to high residue and persistence, easy accumulation and strong harmful effects. The National Research Council of the United States of America published Toxicity Testing in the 21st Century: A Vision and Strategy in 2007, which proposed a new concept of toxicity testing that toxicity testing should take full consideration of population exposure data and base on in vitro tests, human cell lines, toxicity pathways and high-throughput screening. Meanwhile, systems biology, bioinformatics and rapid assay technologies will be used to better understand toxicity pathways-the cellular response pathways that can lead to adverse health effects when sufficient perturbing induced by chemicals exposure. The new toxicity testing strategy has changed the traditional testing pattern and has brought a wide impact on the international relevant fields. The European Union, the World Health Organization, and the United States Environmental Protection Agency, the Food and Drug Administration, and the National Center for Toxicological Research have organized relevant discussions and exploratory studies to address the new toxicity testing concept and how to evaluate and utilize the results of traditional toxicity test researches. Compared to the discussion, 'whether to do it', ten years ago, the question, 'how to do it', has become the concern of the current discussion. Therefore, how to respond to the concept of toxicity testing and how to effectively utilize and excavate traditional toxicity test data have been the focus of multi-disciplines and interdisciplinary academia such as toxicology, food hygiene and environmental science. Therefore, this article provides an overview of the exposure levels of dioxin, polybrominated diphenyl ethers and benzo[a]pyrene, which are typical persistent organic pollutants in food in China and the current research status of toxic pathways based on whole animal experiments. The exposure level, toxic effect and toxicity mechanism of three contaminants are analyzed and summarized in order to provide basis for future results based on the 21st century toxicity test compared with traditional tests and data mining analysis of these two kinds of data. Meanwhile, it also lays the foundation for the establishment of a toxicity testing framework based on exposure characteristics, toxic pathways, and biomarkers.

[Management and safety assessment of nanomaterials in food: status and prospects].

The safety assessment of nanomaterials in food is essential for safeguarding supervision and maintaining public health. However, there are still no safety assessment procedures for nanomaterials established in national-level in China and no specific toxicology and safety assessment procedures about nanomaterials for food, too. These factors lead to restriction on food safety protection and supervision. Current methods of evaluating the safety of nanomaterials mainly rely on traditional toxicological assessment that are extrapolated based on animal experiment from high doses to low doses and from animals to humans. These uncertainties restrict the accuracy of safety assessment for nanomaterials and also limit the development of scientific and effective evaluation procedures and regulatory measures. Currently, the key issues need to be solved including exposure assessment and evaluation methods of nanomaterials in food and the established methods of the toxicity test for nanomaterials that are consistent with the objectives of toxicity test in the 21st century vision and strategy. In this article, we reviewed current administrative regulatory, situations, and existing issues of food nanomaterials either in China or some developed countries in order to provide a scientific basis in establishing safety assessment procedures for nanomaterials in food in the future.

[Viability of osteoblasts under cell hypoxia condition].

Objective: To investigate the effects of hypoxia condition and hypoxia-reoxygenation condition on the cell viability, apoptosis rate and gene expression of osteoblasts cultured in vitro. Methods: The cranium osteoblasts from newborn Sprague Dawley rats within 48 hours were cultured and purified through tissue block method.The morphological changes of cells were evaluated by the Alizarin Red S staining and Alkaline phosphatase staining.The third-generation osteoblasts were cultured in normal condition for 36 hours (group A), in hypoxic condition for 24hours (group B), in hypoxic condition for 24hours thereafter reoxygenated for 12 hours (group D), in hypoxic condition for 36 hours (group C). The cell viability of osteoblasts was tested via MTT assay.The apoptosis rate of osteoblasts was tested by FCM (flow cytometry). Quantitative PCR and Western blot methods were used to determine Collagen type Ⅰ, Bone morphogenetic protein 2 (BMP-2), Runt-related transcription factor 2 (RUNX-2), Transforming growth factor-ß1(TGF-ß1) expression levels. Results: The cell viability of osteoblasts decreased, group A(99.1%±8.3%), group B(90.9%±9.4%), group C(79.9%±8.7%), group D(73.0%±8.2%), group D group C >group B >group A(F=26.198, P=0.000); the mRNA expressions of Col Ⅰ, BMP-2, RUNX-2, TGF-ß1 decreased under hypoxia and hypoxia-reoxygenation condition, and group D< group C< group B< group A (F=13.082, P=0.006; F=7.088, P=0.017; F=6.857, P=0.038; F=51.368, P=0.000); the protein expressions of Col Ⅰ, BMP-2, RUNX-2, TGF-ß1 decreased under hypoxia and hypoxia-reoxygenation condition, and group D< group C< group B< group A (F=8.114, P=0.013; F=28.935, P=0.000; F=9.857, P=0.007; F=46.541, P=0.000). Conclusion: Hypoxia condition and hypoxia-reoxygenation condition decreased the cell viability, increase the apoptosis rate and suppress the expressions of associated genes.Hypoxia-reoxygenation condition aggravate the injure of osteoblasts preconditioning under hypoxia condition.

[Quantitative structure-activity relationship prediction of carcinogenicity of N-nitroso compounds based on category approach and read-across].

Objective: New quantitative structure-activity relationship (QSAR) method was used to predict N-nitroso compounds (NOCs) carcinogenicity. This could provide evidences for health risk assessment of the chemicals. Methods: Total 74 chemical substances of NOCs were included as target chemicals for this validation study by using QSAR Toolbox based on category approach and read-across. The included 74 NOCs were categorized and subcategorized respectively using "Organic functional groups, Norbert Haider " profiler and "DNA binding by OASIS V.1.1" profiler. Carcinogenicity of rat were used as target of prediction, the carcinogenicity results: of analogues in chemical categories were cross-read to obtain the carcinogenic predictive results of the target chemicals. Results 74 NOCs included 26 nonclic N-nitrosamines, 24 cyclic N-nitrosamines and 24 N-nitrosamides The sensitivity, specificity and concordance of the category approach and read-across for predicting carcinogenicity of 74 NOCs were 75% (48/64), 70%(7/10) and 74% (55/74) respectively. The concordance for noncyclic N-nitrosamines, cyclic N-nitrosamines and N-nitrosamides were 88% (23/26), 71% (17/24) and 63% (15/24) respectively. Conclusion: QSAR based on category approach and read-across is good for prediction of NOCs carcinogenicity, and can be used for high-throughput qualitative prediction of NOCs carcinogenicity.

Association between dietary intake of folate and MTHFR and MTR genotype with risk of breast cancer.

We investigated the association between dietary intake of folate, vitamin B6, and the 5,10-methylenetetrahydrofolate reductase (MTHFR) genotype with breast cancer. A matched case-control study was conducted, and 413 patients with newly diagnosed and histologically confirmed breast cancer and 436 controls were recruited. Folate intake, vitamin B6, and vitamin B12 levels were calculated, and the MTHFR C677T and A1298C and MTR A2756G polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Breast cancer cases were generally older, older at first live birth, and younger at menarche, had a higher body mass index, were smokers, had higher energy intake, and more first-degree relatives with breast cancer as well as more live births compared to controls. With respect to energy intake, we found that higher energy intake were more likely to increase the risk of breast cancer. The MTHFR 667TT genotype was associated with a moderately increased risk of breast cancer when compared with the CC genotype, and a significant odds ratio (OR; 95% confidence interval, CI) was found (OR = 1.70, 95%CI = 1.06-2.73). Individuals carrying T allele were associated with higher risk of breast cancer when compared with C allele (OR = 1.34, 95%CI = 1.06-1.70). We did not find a significant effect of the MTHFR A1298C and MTR A2756G on the risk of breast cancer. We did not find any association between folate intake and MTHFR C677T polymorphisms. In conclusion, we found that the MTHFR C667T polymorphism is associated with the risk of breast cancer, indicating that this genotype plays a role in breast cancer development.

Clinical significance of CA 19.9 and LINC01197 in pancreatic cancer.

OBJECTIVE: This study aimed to explore the expression and clinical significance of LINC01197 in serum of patients with pancreatic cancer (PC). PATIENTS AND METHODS: Fifty PC patients (patient group) treated in our hospital from March 2012 to April 2014 were collected, and another 50 normal people (normal group) were collected for physical examination. The LINC01197 expression in serum of the two groups was detected by qRT-PCR method, and the CA 19.9 expression in serum was detected by Roche automatic biochemistry. The expression and diagnostic value of CA 19.9 and LINC01197 in PC were analyzed, and the relationship between LINC01197 and prognosis of PC patients was observed. RESULTS: The CA 19.9 expression in the patient group was significantly higher than that in the normal group (p<0.001). Their area under the curve was 0.791 and 0.944 respectively. The incidence of phases III+IV, lymphatic invasion, and distant metastasis in patients with low expression of LINC01197 is significantly higher than that in those with high expression, and has higher diagnostic value. With the progress of clinical staging, the TNM expression decreased gradually and there were differences between groups (p<0.001). Spearman's test analysis found that the decreased TNM staging of LINC01197 increased gradually (r=-0.816, p<0.001), and the area under the curve of LINC01197 distinguishing phase I and phase II+phase+III+phase IV was 0.930. The 1-year survival rate and 5-year survival rate of patients in low expression group were lower than those in high expression group (p1 year =0.037, p5 year =0.014). Distant metastasis is an independent prognostic factor for PC patients to survive for 1 to 5 years. Differentiation, TNM staging, and LINC01197 are independent prognostic factors for PC patients to survive for 5 years. CONCLUSIONS: The low expression of LINC01197 in PC patients indicates poor prognosis of patients and is expected to be a potential diagnostic and prognostic indicator of PC.

MiR-150 alleviates EMT and cell invasion of colorectal cancer through targeting Gli1.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-150 alleviates EMT and cell invasion of colorectal cancer through targeting Gli1, by H. Fan, X. Liu, W.-W. Zheng, Z.-H. Zhuang, C.-D. Wang, published in Eur Rev Med Pharmacol Sci 2017; 21 (21): 4853-4859-PMID: 29164577" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/13726.

MiR-150 alleviates EMT and cell invasion of colorectal cancer through targeting Gli1.

OBJECTIVE: Epithelial-mesenchymal transition (EMT) is related to colorectal cancer invasion and metastasis. Glioma-associated oncogene homolog 1 (Gli1) abnormal expression is associated with EMT, invasion, and metastasis in various cancers. MiR-150 is found downregulated in colorectal cancer pathogenesis. Bioinformatics analysis shows the complementary targeted relationship between miR-150 and the 3'-UTR of Gli1 mRNA. This study explores the role of miR-150 in regulating Gli1 expression, colorectal cancer cell EMT, and invasion. MATERIALS AND METHODS: Dual luciferase assay confirmed the targeted relationship between miR-150 and Gli1 predicted by bioinformatics analysis. MiR-150 and Gli1 expressions were compared in NCM460, SW480, and SW620 cells. Cell colony formation and invasion were tested in SW480 and SW620 cells. Anip973 and AGYZ83-a cells were treated by 10 ng/mL TGF-ß1 to detect miR-150 and Gli1 expressions. SW620 cells were cultured in vitro and divided into five groups, including miR-NC, miR-150 mimic, si-NC, si-Gli1, and miR-150 mimic + si-Gli1 groups. RESULTS: MiR-150 specifically inhibited Gli1 expression. The level of miR-150 was significantly downregulated, while Gli1 was elevated in SW480 and SW620 cells compared with that in NCM460 cells. SW620 exhibited markedly stronger invasive and colony formation abilities than SW480. The level of miR-150 was apparently reduced, whereas Gli1 was increased in SW620 than that in SW480 cells after the treatment of TGFß1. MiR-150 mimic and/or si-Gli1 transfection markedly reduced Gli1 and Snail levels, upregulated E-cadherin expression, and attenuated cell colony formation and invasion. CONCLUSIONS: Downregulation of miR-150 and elevation of Gli1 promote the development and invasion of colorectal cancer cell EMT. MiR-150 attenuated the progression of colorectal cancer cell EMT via inhibiting Gli1.

MicroRNA-216b is downregulated in hepatocellular carcinoma and inhibits HepG2 cell growth by targeting Forkhead box protein M1.

OBJECTIVE: Aberrantly expressed microRNAs (miRs) may play critical roles in the regulation of tumorigenicity of various cancers. The present study was designed to investigate the expression, function and the underlying mechanism of miR-216b in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The expression of miR-216b and FOXM1 in 24 paired HCC tissues and adjacent normal tissues was determined by Real-time PCR. The proliferative activity of HepG2 cells was determined by MTT assay. We analyzed cell cycle progression by flow cytometry, apoptosis by cell death enzyme-linked immunosorbent assay (ELISA) and cleaved-caspase-3 by western blot. Luciferase reporter assay was employed to verify whether FOXM1 serves as a target of miR-216b in vitro. RESULTS: The expression of miR-216b was significantly decreased in HCC tissues compared with that in adjacent normal tissues, whereas FOXM1 expression was increased. In addition, FOXM1 and miR-216b expression were inversely correlated in HCC tissues. Ectopic expression of miR-216b produced a suppressive effect on the growth of HepG2 cells and induced cell cycle arrest and apoptosis. We further demonstrated that miR-216b targets the 3' untranslated region (UTR) of FOXM1 directly to suppress the expression of FOXM1, and that suppression of FOXM1 produced the similar effects to miR-216b CONCLUSIONS: These data suggest that down-regulation of miR-216b directly contributes to the up-regulation of FOXM1, which may confer the tumorigenicity of HCC cells. MiR-216b may serve as a potential therapeutic agent for HCC.

[Chronic neutrophilic leukemia complicated with multiple myeloma: two cases report and literature review].

OBJECTIVE: To explored the diagnosis and treatment of chronic neutrophilic leukemia (CNL) complicated with multiple myeloma (MM). METHODS: The clinical features and molecular biological characteristics of 2 patients with CNL complicated with MM were summarized, and the diagnosis and treatment of the patients were retrospectively reviewed. RESULTS: The diagnosis of CNL complicated with MM was established in 2 cases. Case 1 had CSF3R mutation (P733T), but CSF3R-exon 14 mutation and SETBP1 mutation were all negative. The neutrophil count returned to normal when MM was successfully treated in case 1. When the patient relapsed, neutrophil count increased again. CONCLUSION: Coexistence of CNL and MM is rare. CSF3R is a very important molecular marker for CNL. To the best of our knowledge, it's the first time to report the coexistence of CNL and MM carried CSF3R mutation (P733T). Chemotherapy regimens for MM may be effective in the treatment of CNL complicated with MM.

Cochlear gene delivery through an intact round window membrane in mouse.

Cochlear gene transfer studies in animal models have utilized mainly two delivery methods: direct injection through the round window membrane (RWM) or intracochlear infusion through a cochleostomy. However, the surgical trauma, inflammation, and hearing loss associated with these methods lead us to investigate a less invasive delivery method. Herein, we studied the feasibility of a vector transgene-soaked gelatin sponge, Gelfoam, for transgene delivery into the mouse cochlea through an intact RWM. The Gelfoam absorbed with liposomes and adenovirus, but not with adeno-associated virus (AAV), was successful in mediating transgene expression across an intact RWM in a variety of cochlear tissues. The Gelfoam technique proved to be an easy, atraumatic, and effective, but vector-dependent, method of delivering transgenes through an intact RWM. Compared with the more invasive gene delivery methods, this technique represents a safer and a more clinically viable route of cochlear gene delivery in humans.

Cationic liposome mediated transgene expression in the guinea pig cochlea.

Sensorineural hearing loss affects nearly 10% of the American population that is refractory to conventional therapy. Gene therapy represents an intervention with potential therapeutic efficacy. We studied the feasibility of cationic liposome mediated gene transfer within the guinea pig cochlea in vivo following direct microinjection into the cochlea. Transgene expression was persistent up to 14 days in the neurosensory epithelia and surrounding tissue without toxicity and inflammation in the target organ. This study represents the first successful use of cationic liposomes for cochlear gene transfer thus providing a safe and rapid alternative to the use of recombinant viral vectors in gene therapy for inner ear disorders.

Genospheres: self-assembling nucleic acid-lipid nanoparticles suitable for targeted gene delivery.

We describe the assembly of a cationic lipid-nucleic acid nanoparticle from a liquid monophase containing water and a water miscible organic solvent where both lipid and DNA components are separately soluble prior to their combination. Upon removal of the organic solvent, stable and homogenously sized (70-100 nm) lipid-nucleic acid nanoparticles (Genospheres) were formed. The low accessibility (<15%) of the nanoparticle-encapsulated DNA to a DNA intercalating dye indicated well-protected nucleic acids and high DNA incorporation efficiencies. It was demonstrated that Genospheres could be stably stored under a variety of conditions including a lyophilized state where no appreciable increase in particle size or DNA accessibility was observed following reconstitution. Finally, Genospheres were made target-specific by insertion of an antibody-lipopolymer (anti-HER2 scFv (F5)-PEG-DSPE) conjugate into the particle. The target specificity (>100-fold) in HER2 overexpressing SK-BR-3 breast cancer cells was dependent on the degree of PEGylation, where the incorporation of high amounts of PEG-lipid on the particle surface (up to 5 mol%) had only a minor effect on the transfection activity of the targeted Genospheres. In summary, this work describes a novel, readily scalable method for preparing highly stable immunotargeted nucleic acid delivery vehicles capable of achieving a high degree of specific transfection activity.

Increased target specificity of anti-HER2 genospheres by modification of surface charge and degree of PEGylation.

Genospheres are cationic lipid-nucleic acid nanoparticles prepared by the assembly of the lipids and nucleic acids from an aqueous/organic liquid monophase that independently dissolves the components, where the resultant particles are homogeneously sized (70-110 nm), with efficiently incorporated and protected DNA. In the present study, we demonstrate pH-dependent modulation of the Genosphere surface charge using pH-titratable lipids. By incorporation of the lipids with titratable anionic or imidazole headgroups, Genospheres with neutral or anionic surface charge at neutral pH were produced and compared for cellular uptake and transfection of a reporter gene (luciferase) in culture of breast cancer cells. The extent of particle-cell association was also studied by fluorescent microscopy and quantified by cytofluorometery. The effects of Genosphere surface modification with poly(ethylene glycol) (molecular weight 2000) at low (0.5 mol %) and high (5 mol %) grafting densities, as well as the effects of HER2-receptor-directed targeting by an internalizable anti-HER2 scFv F5, linked via PEG spacer, were also studied. Inclusion in the Genosphere formulation of pH-titratable lipids CHEMS (cholesteryl hemisuccinate), CHIM (1-(3-(cholesteryloxycarbonylamino)propyl)imidazole), or DSGG (1,2-distearoyl-sn-glycero-3-hemiglutarate) rendered the particles surface-charge neutral or slightly anionic at neutral pH, and cationic at mildly acidic pH, as shown by zeta-potential measurements. In HER2-targeted systems, transfection activity and target specificity with HER2-overexpressing SKBR-3 breast cancer cells were dependent on Genosphere surface charge and PEGylation. The highest target specificity correlated with low cationic charge at neutral pH, while incorporation of 5 mol % PEG-lipid had only minor effects on Genosphere-cell association, internalization, and transfection activity. The implications of this work for potential in vivo applications are discussed.