[Role of LINC00310 in the glycidyl methacrylate-induced malignant transformation of human bronchial epithelial cell].

OBJECTIVE: To investigate the expression and biological significance of LINC00310 in the malignant transformation of human bronchial epithelial cells(16 HBE) induced by glycidyl methacrylate(GMA). METHODS: The 16 HBE cells recovered successfully used 1 µg/mL dimethyl sulfoxide(DMSO) as the solvent control group, and the final concentration was 8 µg/mL GMA as the treatment group, and were subcultured after repeated exposure 3 times for 72 hours each time. The 10 th, 20 th and 30 th generation cells of the GMA treatment group and corresponding DMSO control group were collected. The LncRNA microarrays was used to analyze the expression changes of LINC00310 in different periods, and the target gene and function prediction was performed by NCBI and cBioPortal bioinformatics database, and real-time quantification PCR(qPCR) was used to detect the relative expression levels of LINC00310 and predicted target genes. RESULTS: The result of the microarray showed that LINC00310 in the GMA-treated group was down-regulated by 2. 02-fold, up-regulated by 6. 17-fold, and up-regulated by 2. 03-fold in the pre-transformation, mid-term, and late, respectively. The result of qPCR confirmed that the expression of LINC00310 relative expression level of 10 th, 20 th and 30 th generation cells was consistent with the microarray result, which were down-regulated by 2. 76-fold, up-regulated by 2. 68-fold, and up-regulated by 3. 09-fold. Consistently, the relative expression of the target gene C-Myc was statistically significant in 20 th and 30 th generation cells. CONCLUSION: LINC00310 induced low expression in the early stage of malignant transformation of 16 HBE cells induced by GMA, and was highly expressed in the middle and late stages. It indicated that LINC00310 may play a cancer-promoting role in the process of cell malignant transformation through C-Myc.

[Anti-oncogene of opioid binding protein/cell adhesion molecule-like methylation status in malignant transformation of human bronchial epithelial cells induced by glycidyl methacrylate].

OBJECTIVE: To analyze the opioid binding protein/cell adhesion molecule-like (OPCML) methylation status at different stages of malignant transformation of human bronchial epithelial (16HBE) cells induced by Glycidyl Methacrylate (GMA) and to explore the effect of OPCML methylation in the process of malignant transformation. METHODS: Cells were harvested at different stages (the 10th generation, the 20th generation and the 30th generation). To verify the Methylation chip result of OPCML methylation status in the process of malignant transformation, we detected it by methylation-specific-PCR (MSP); Real-time fluorescence Quantitative PCR (qPCR) were applied to measure the gene expression levels of OPCML at different transformed stage, and compared with the control groups (treated with DMSO). RESULTS: Based on the result of methylation chip, the gene of OPCML methylation occurred in all stages, which was consistent to the result of MSP; qPCR showed that the levels of gene expression decreased in the 20th generation and 30th generation. CONCLUSION: Methylation status of OPCML gene promoter could be considered as a stable and specific biomarker in the transformation process.

Interleukin 1ß and interleukin 1 receptor antagonist gene polymorphisms and cervical cancer: a meta-analysis.

OBJECTIVES: Previous studies investigating the association between interleukin 1ß (IL-1ß) and its receptor antagonist (IL-1RN) polymorphism and cervical cancer risk have reported controversial results. Thus, we examined these associations by performing meta-analyses. METHODS AND MATERIALS: Fourteen studies testing the association between IL-1ß and/or IL-1RN gene polymorphisms and cervical cancer were examined: 5 studies of IL-1ß-511C/T, 3 studies of IL-1ß-31T/C, and 6 studies of IL-1RN. Overall and ethnicity-specific summary odds ratios (ORs) and corresponding 95% confidence intervals (CIs) for cervical cancer associated with these polymorphisms were estimated using fixed- and random-effects models. Heterogeneity and publication bias were evaluated. RESULTS: Meta-analysis of all 6 studies showed variant genotypes of IL-1RN to be associated with an elevated cervical cancer risk (RN2/RN2 vs RN1/RN1: OR, 2.64; 95% CI, 1.29-5.40; recessive: OR, 2.15; 95% CI, 1.06-4.38; dominant: OR, 1.60; 95% CI, 1.07-2.38). Combined analysis indicated that IL-1ß-511C/T polymorphism was also associated with increased risk of cervical cancer (TT vs CC: OR, 1.56; 95% CI, 1.22-1.99; CT vs CC: OR, 1.61; 95% CI, 1.31-1.99; dominant: OR, 1.60; 95% CI, 1.31-1.95). No significant association of IL-1ß-31T/C and cervical cancer risk was detected. There was no evidence of publication bias. CONCLUSIONS: This meta-analysis suggested that the IL-1RN and IL-1ß-511C/T polymorphisms may contribute to genetic susceptibility of cervical cancer. More studies are needed to further evaluate the role of the IL-1ß-31T/C polymorphism in the etiology of cancer.

[Determination of 2, 4-dichlorophenoxyacetic acid in air of workplace by high-performance liquid chromatography].

OBJECTIVE: To develop a method for determination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) in the air of workplace by high-performance liquid chromatography. METHODS: 2, 4-D was collected by ultrafine glass filters, desorbed by methanol, separated by a C18 column, and detected by a UV detector. Identification and quantification of 2, 4-D were performed by retention time and peak areas, respectively. RESULTS: The linear range of the test was 2∼200 µg/ml; the elution efficiency was 94.6%- 95.9%; the limit of detection (S/N = 3) was 0.034 µg/ml (injection volume of 20 µl eluant); the lower limit of quantification (S/N = 10) was 0.11 µg/ml; the minimum detectable concentration was 0.011 mg/m(3); the minimum quantifiable concentration was 0.037 mg/m(3) (with sampled air volume of 45 L). CONCLUSION: This method is convenient and simple in sample collection and preparation, and satisfies all methodological requirements. Therefore, this method is useful for the determination of 2, 4-D in the air of workplace.

[Effect of Huperzine A on neural lesion of acute organophosphate poisoning in mice].

OBJECTIVE: Effects of neurophathologic changes and expression of Glu and 60 nNOS were observed in acute isocarbophos and phoxim poisoning in mice. METHODS: KM male mice were randomly divided into three groups, which were control, non-treated and Huperzine A (HupA)-treated groups. The control group was given tween-80. Nontreated group was given isocarbophos (14.7 mg/kg) or phoxim (1702 mg/kg). HupA-treated group was given HupA 2h before phoxim or isocarbophos. Twenty-four hours after exposure, the whole brain was removed and adjacent coronal sections was obtained. One part of sections were stained with toluidine blue. The part of sections were used to assessed the expression of Glu and nNOS in the cortex and hippocampal of brain by immunohistochemistry. RESULTS: Compared to control group, non-treated group was observed nissal body nembers reduced and dyeing light. The animals of HupA protective group were observed nissal body nembers reduced, but the lesional degree was lighter obviously than non-treated group. The statistically reduced of the expression of Glu (P<0.01), the elevation of nNOS (P<0.01), after Isocarbophos intoxication were observed. Compared to non-treated group, the significant elevation of Glu (P<0.01) and reduced of nNOS (P<0.01) was observed on HupA-treated groups. Whereas for phoxim treatment, no changes were observed. CONCLUSION: HupA have protective effect against glutamatergic systems disorder caused by Isocarbophos poisoning. Administration of HupA have no effects of the neurotransmitter changes induces by acute poisoning of phoxim. It is different for the toxic effect mechanism of the two organophosphate.

[Oxidative damage effects induced by CdTe quantum dots in mice liver].

OBJECTIVE: To investigate oxidative damage effects induced by CdTe quantum dots (QDs) in mice liver. METHODS: 40 ICR mice were randomly divided into 5 groups, one control group (normal saline), four CdTe QDs (exposed by intravenous injection of 0.2 ml CdTe QDs at the concentration of 0, 0.5, 5, 50 and 500 nmol/ml respectively). All mice were decapitated 24h after the injection. Concentration of MDA and the activities of SOD, CAT, 8-OHdG expression were examined by immunohistochemistry and hepatocellular apoptosis was measured with TUNEL resperctively. RESULTS: The results showed in 500 nmol/ml CdTe QDs group, MDA concentration in the liver tissue were significantly higher than those of control group (P < 0.05). The activities of SOD in 50 and 500 nmol/ml and CAT in 5, 50 and 500 nmol/ml CdTe QDs groups were significantly lower than those of control group (P < 0.01). Positive staining for 8-OHdG appeared in 50 and 500 nmol/ml CdTe QDs groups and the apoptosis of liver cells was observed in 500 nmol/ml CdTe QDs group by TUNEL technique. CONCLUSION: It was suggested that CdTe QDs at certain doses could induce oxidative damage effects of the mice liver in the condition of this experiment.

[Effects of CdTe QDs on chromosome aberration of CHL cells in vitro].

OBJECTIVE: To observed the chromosome oberration of CHL cells induced by CdTe QDs. METHODS: The chromosome oberration test of Chinese hamster lung (CHL) cell was conducted in CdTe QDs at the concentrations of 1.6, 3.2, 6.3, 12.6 and 25.2 microl/ml, under the metabolic (+S9) and non-metabolic (-S9) activation systems. RESULTS: The chromosome oberration rate of 12.6 and 25.2 microl/ml CdTe QDs groups were significantly higher than the control groups (P<0.01) Under the condition of metabolic activation. Main types of chromosomal aberrations was broken, fragmented and exchange. CONCLUSION: It was suggested that CdTe QDs could induce the effects of chromosome oberration under the metabolic activation systems (+S9).

[Effects of CdTe QDs on oxidative stress and DNA damage of liver cells in mice].

OBJECTIVE: To investigate the effects of CdTe QDs (cadmium telluride quantum dots) on oxidative stress and DNA damage of liver cells in mice. METHODS: Thirty ICR male mice were randomly divided into 5 groups: one negative control (normal saline) group. Three CdTe QDs groups (exposed by intravenous injection of 0.2 ml of CdTe QDs at the concentration of 3.75, 37.5 and 375 nmol/ml respectively) for electron paramagnetic resonance (EPR) test, and another positive control group (exposed by intravenous injection of 0.2 ml of cyclophosphamide 20 mg/ml) for single cell gel electrophoresis (SCGE) test. All mice were decapitated 24h after the injection, free radicals and DNA damage of liver cells were detected by EPR and SCGE. RESULTS: The levels of oxygen free radicals detected by EPR were increased with the increase of CdTe QDs. The tail length, olive tail moment, tail DNA (%) and the ratio of tail/head examined by SCGE were also increased with the increase of the dosage of CdTe QDs (P < 0.01). CONCLUSION: CdTe QDs could induce oxidative stress and DNA damage of liver cells in mice with a dose-effect relationship.

[Chromosome damage of human bronchial epithelial cells induced by glycidyl methacrylate].

OBJECTIVE: To observed the chromosome damage of human bronchial epithelial cells induced by glycidyl methacrylate (GMA). METHODS: Chromosome aberration analysis of human bronchial epithelial cells treated with GMA at different dosages (4, 8, 12, 16 and 20 microg/ml), times (1, 2 and 3 times), and phases (10th, 30th genetation) was detected. RESULTS: In the dosages range from 4 to 20 microg/ml, the aberration rates (3%, 6%, 7%, 11% and 14%) were demonstrated increasingly with the increase of exposure doses, and dose-effect relationship was found. Significant differences were observed when treated with GMA three times (6%, 7% and 10%). Structure aberrations were found in the transformed 10th-generation cells, while number aberrations were mainly manifested in the transformed 30th-generation cells. CONCLUSION: The chromosome aberration can be induced by GMA in the human bronchial epithelial cells, from the structure aberration at the beginning to the lack of normal nuclear style.

[Changes of pathologic feature and microtubulin associated protein 2 in nervous system of hens with organophosphate-induced delayed neuropathy induced by 2,4,6-trimethylbenzoyl phenylphosphonate].

OBJECTIVE: To develop the organophosphate-induced delayed neuropathy (OPIDN) hen model with 2,4,6-trimethylbenzoyl phenylphosphonate (TOCP), and observe the change of pathology and investigate the alterations of microtubulin associated protein 2 (MAP2). METHODS: 48 adult hens were randomly divided into four groups, including three experimental groups and control group (n = 12 each group). The hens in three experimental groups were treated with TOCP by gavage at single dosages of 250, 500 and 750 mg/kg respectively while the control hens received an equivalent volume of corn oil by gavage. All hens were sacrificed after 21 days of treatment. Half hens in each group were dissected for HE examination and myelin straining of brain, spinal cord and sciatic nerve while brains of another half hens were dissected for the determination of MAP2 by western blotting. RESULTS: The delayed neurotoxicity symptoms of hens both in 500 and 750 mg/kg groups were consistently observed. The pathological changes of brain, spinal cord and sciatic nerve in 500 and 750 mg/kg groups showed nerve cells difference necrosis, increased cytoplasm basophilia, microglia proliferation, mono-nuclear and lymphocyte infiltration, myelin sheath extensive up to part of them disaggregation deletion. Compared with the control group, at 500 and 750 mg/kg respectively the increase of MAP2 was 25% and 23% (P < 0.01 and P < 0.05). CONCLUSIONS: The histopathologic changes of OPIDN caused by TOCP have dose-response relationship. The changes of MAP2 in nervous system may contribute to the occurrence and development of TOCP induced delayed neurotoxicity.

[Study on malignant transformation of human bronchial epithelial cells induced by glycidyl methacrylate].

OBJECTIVE: To study the malignant transformation of human bronchial epithelial cells induced by glycidyl methacrylate (GMA). METHODS: 16HBE cells were treated multiple times with GMA at concentrations of 1, 2, 4 and 8 microg/ml. Cellular biological characteristics of malignant transformation were identified by the tests of conA, colony forming frequency on soft agar, scanning electron microscope and tumorigenesis in nude mice. Test of immunocytochemical detection was also applied to confirm the derivation of cell and tumor. Groups of solvent control (DMSO) and positive control (MCA) were also performed at the same time. RESULTS: Transformed foci could be observed after the cells were treated by GMA at concentrations from 1 to 8 microg/ml. The number of transformation foci increased with the concentration of GMA. Transforming rate in 8 microg/ml group (8.48 x 10(-6)) was significantly higher (P < 0.01) than that of solvent control group (4.5 x 10(-7)). The transformed cells lost contact inhibition and exhibited a crossover growth in culture dish. They also could grow in semi-solid agar and showed dose-reaction relations with the concentration of GMA. The colony forming frequency in 2, 4 and 8 microg/ml group was 1.20 per thousand, 2.35 per thousand and 5.70 per thousand respectively, which were higher than that of solvent control group (P < 0.01). The transformed cells could be agglutinated by low concentration of conA. Microvilli on the surface of transformed cells increased and became strong and long under scanning electron microscope. The transformed cells could form subcutaneous tumor in nude mice which was diagnosed as squamous cell carcinoma in morphology. Expression of cytokeratin (CK) was detected in both 16HBE cells and tumor formed in nude mice. CONCLUSION: GMA could induce the malignant transformation of 16HBE cells. This research system might provide a potential tool and lay a foundation for the study of the molecular mechanism of carcinogenesis induced by GMA.

[Effect of HuperzineA on neurophathologic change of mouse exposed by isocarbophos and phoxim].

OBJECTIVE: In order to assessment the Huperzine A (HupA) on CNS of mice exposed by phoxim and isocarbophos. METHODS: One group was given tween-80 or peanut oil as control. One group was given phoxim or isocarbophos alone as phoxim or isocarbophos group. The other group was given HupA 2h before phoxim or isocarbophos exposure as HupA protective group. Twenty-four hours after exposure, the whole body perfusion in situ technique was performed for tissue fixation. The whole brain was removed and adjacent coronal sections was obtained. The sections of brain were stained with hematoxylin and eosin (HE) and toluidine blue. Under optic microscope images were collected and analysed. RESULTS: The extensive lesions including cortex oedema, pyramidal cell oedema, and nissal body numbers reduced were observed in phoxim or isocarbophos group. The animals of HupA protective group were observed cortex oedema, pyramidal cell oedema, but the lesional degree was lighter obviously than phoxim or isocarbophos group. CONCLUSION: It was suggested that the HupA could have the protective effect of neuropathologic changes exposed by phoxim and isocarbophos.

[Experimental study on protective effects of HupA in the treatment of isocarbophos poisoning].

OBJECTIVE: To investigate the therapeutic and prophylactic efficiency of HupA in mice with acute isocarbophos poisoning, and the protective effects of the HupA on AChE inhibited by isocarbophos. METHODS: Mice were randomizedly divided into the non-treatment group, the atropine control group, the HupA treatment group and the atropine and HupA combined treatment group. Toxic signs and survival rates were observed and compared among these groups. The AChE activity was monitored in the whole blood, the red cells and brain tissue exposed to isocarbophos in the either treated with HupA or non-treated groups. RESULTS: In HupA treatment group compared with the non-treatment group, toxic signs were significantly decreased and the survival rate was increased. The therapeutic efficiency in the atropine and HupA combined treatment group was better than other groups. After isocarbophos was administered, the AChE activity in the HupA treatment group and the non-treatment group was decreased. However, the AChE activity in the whole blood (1.096 +/- 0.111), (1.262 +/- 0.146), (1.181 +/- 0.353) U/ml, the red cells (0.798 +/- 0.063), (1.000 +/- 0.176), (0.837 +/- 0.331) and the brain tissue (13.739 +/- 2.970), (18.507 +/- 3.466), (10.764 +/- 2.212) U/g in HupA treatment group 0.5, 1 and 2 hours after isocarbophos was administered was significantly higher than those in the non-treatment group (P < 0.05 or P < 0.01). CONCLUSION: HupA has therapeutic effect on mice with acute isocarbophos poisoning. The protective effect of HupA on blood and brain AChE inhibited by isocarbophos may be one of the mechanisms of the therapeutic effect of HupA in acute Isocarbophos poisoning.

[The polymorphism of delta-aminolevulinic acid dehydratase and genetic susceptibility to lead toxicity in Uighur and Yi population in China].

OBJECTIVE: To investigate the polymorphism of delta-aminolevulinic acid dehydratase(ALAD) and the genetic susceptibility to lead toxicity in Uighur and Yi population in China. METHODS: The ALAD genotypes were determined by PCR and MspI restriction fragment length polymorphism techniques in 214 Uighur individuals from Xinjiang autonomous region and 144 Yi individuals from Yunnan province. The correlation between the polymorphism of ALAD and blood lead levels, and the factors affecting the latter were explored. RESULTS: The frequencies of the allele ALAD1 and ALAD2 in Uighur are 0.91 and 0.09; and in Yi are 0.98 and 0.02 respectively. In Uighur the average blood lead level was (76 +/- 4) microgram/L, and 25.7% individuals with blood lead level > or = 100 micrograms/L. In Yi the average blood lead level was (50 +/- 16) microgram/L, and 6.3% individuals with blood lead level > or = 100 micrograms/L. However, no statistic correlation between the distribution of ALAD alleles and the blood lead level was found in both populations. CONCLUSION: The genetic susceptibility of ALAD polymorphism to lead toxicity may exhibit in a lead dose-dependent manner.