OBJECTIVE:This study aimed to explore the copy number variations and expression of MPDZ and to evaluate their relationship with prognostic outcomes in liver cancer. METHODS:Copy number variations and expression of MPDZ were performed from TCGA database,and their relationships were analyzed. Expression of MPDZ in liver carcinoma (282 cases) and adjacent normal tissues (50 cases) were also performed,then the relationship between MPDZ expression and clinical pathological parameters,and its prognostic value were evaluated by statistical analysis. RESULTS:Copy number deletion (31.44%) was the most frequent copy number variations observed and it was associated with the expression of MPDZ. The expression was significantly lower than that in adjacent normal tissues (P < 0.05). Kaplan-Meier survival curves showed that patients with low MPDZ expression had significantly shorter overall survival time than those with higher MPDZ expression (Log-rank=12.48,P < 0.05). Receiver operating characteristic curve (ROC) showed that MPDZ gene expression was a sensitive and specific index of liver cancer diagnosis (AUC=0.86). CONCLUSION:MPDZ gene copy number deletion might be associated with the occurrence of liver cancer and the expression of MPDZ had some reference value for the diagnosis and prognosis of liver cancer.

OBJECTIVE:To detect the expression and abundance of different transcripts of KIAA1522 gene in esophageal squamous cell carcinoma (ESCC) tissues and cell lines,and to determine the apparent molecular weight of the protein. METHODS:The expression and abundance of four different transcripts of KIAA1522 gene in ESCC tissues and cell lines were detected by reverse transcription PCR(RT-PCR) and quantitative real-time PCR(qPCR). Molecular weight of the KIAA1522 protein was determined by Western blot together with experiments that involved KIAA1522 gene specific knockdown or exogenous over-expression. The sequence of KIAA1522 protein was confirmed by mass spectrometry. RESULTS:Both T1 and T4 transcripts of KIAA1522 were expressed in ESCC tissues and cell lines,but T1 was the most abundant. The 170 kDa band observed in the Western blot was the KIAA1522 gene-specific protein expression product. CONCLUSION:T1 was the predominant transcript of KIAA1522 gene in ESCC tissues and cell lines. The apparent molecular weight of KIAA1522 protein was 170 kDa.

OBJECTIVE:To investigate the effects of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) on cell proliferation and expression of the main nuclear receptors of RXRα,TRs and PPARs in human neuroblastoma SK-N-SH cells. METHODS:SK-N-SH cells were treated with different concentrations of BDE-47 and their proliferation was measured by the Cell Counting Kit-8 (CCK-8) assay. In addition,ROS level,SOD vitality,GSH-Px vitality and hOGG1 mRNA expression were analyzed by using the DCFH-DA,WST-1,colorimetric and qPCR assays,respectively. Involvement of the main nuclear receptors on expression of mRNA and protein were determined by qPCR and Western blot,respectively. RESULTS:BDE-47 inhibited cell proliferation and induced significant cytotoxicity in SK-N-SH cells. Dose-and time-dependent responses of the inhibitory effect were concentration-dependent,and the IC₅₀ for 24-hours was 75.94 μmol/L. The expression levels in mRNA and protein of RXRα,TRs and PPARs in the treated cells were significantly increased compared with that in the control group,while induction of the isoforms was not consistent. CONCLUSION:BDE-47 inhibited proliferation of SK-N-SH cells via the induction of oxidative damage and neurotoxicity through up-regulating the expression of nuclear receptors:RXRα,TRs,and PPARs.

OBJECTIVE:To explore the effects of high fat combined with iron overload diet in rats on lipid metabolism in the development of nonalcoholic fatty liver disease (NAFLD). METHODS:Forty-eight male SD rats were randomly divided into 5 groups:normal control,high iron diet (high iron diet containing 1% FeSO₄),high fat diet (high fat diet),high fat and low dose iron supplement diet (high fat diet containing 0.5% FeSO₄),high fat and high dosage iron diet (high fat diet containing 1% FeSO₄) groups. These rats were sacrificed after 20 weeks. Their body weights were measured and liver lipid accumulation was observed by oil red O staining. The content of liver TG was measured. Quantitative real-time PCR (qPCR) were used to detect mRNA levels of fatty acid synthase (FAS) and carnitine palmitoyl transferase 1 (CPT1) and their proteins were detected by Western blot. RESULTS:Compared with the normal control group,the body weight and liver TG content in the high fat group were increased significantly (P < 0.05) and high fat diet promoted the deposition of TG in the liver. Additionally,the mRNA level and protein expression of FAS were significantly increased,but the mRNA level and protein expression of CPT1 were significantly decreased (P < 0.05). Compared with the high fat group,the content of liver TG in the high fat and high dosage iron supplement group was significantly increased (P < 0.05). The mRNA and protein level of FAS were increased significantly and the mRNA and protein level of CPT1 were significantly decreased (P < 0.05). CONCLUSION:Iron overload further promoted the deposition of TG in the liver,the disorder of lipid metabolism and the development of NAFLD.

OBJECTIVE:To explore the impact of iron overload on iron metabolism index of Hepcidin and Fpn-1 in HepG2 cells,a model of nonalcoholic fatty liver disease (NAFLD). METHODS:Cell viability was determined using the MTT assay at different concentrations (0.062 5,0.125,0.25,0.5,1.0,2.0 mmol/L,respectively) of oleic acid (OA) and iron and to determine the combined drug concentration. The model of NAFLD was established by using 0.5 mmol/L OA in combination with different concentrations (0,0.125,0.25,0.5 mmol/L) of the Fe induced in HepG2 cells. The contents of intracellular triglyceride (TG) and ferritin (Fn) were determined. The mRNA levels of Hepcidin and Ferroportin1 (Fpn-1) were determined by quantitative real-time PCR and the protein expression of Hepcidin and Fpn-1 were determined by Western blot. A negative control group (no drug treatment of cells) and a 0.5 mmol/L Fe group were used as controls. RESULTS:Compared with the negative control group,0.5 mmol/L OA,0.125,0.25,0.5 mmol/L Fe had no obvious effect on cell viability (P > 0.05). Compared with the control and the 0.5 mmol/L OA groups,in groups of 0.5 mmol/L OA joint with various concentrations of Fe and with increased of the concentration of iron,intracellular TG and ferritin (Fn) content increased gradually and significantly (P < 0.05). Compared with the control and the 0.5 mmol/L OA group,the joint effect of 0.5 mmol/L OA with various concentrations of Fe resulted in the decrease of mRNA and protein expression of Hepcidin (P < 0.05),resulted in the increase of mRNA of Fpn-1 (P < 0.05). Compared with the control,0.5 mmol/L Fe group,0.5 mmol/L OA combined with 0.25 and 0.5 mmol/L Fe resulted in the decrease of protein expression of Fpn-1 (P < 0.05). CONCLUSION:The joint effect of 0.5 mmol/L OA with various concentrations of Fe aggravated the pathogenesis of NAFLD through lipid deposition and iron dysfunction.

OBJECTIVE:To explore the effect of folic acid (FA) on genetic damage and DNA methylation which were induced by 1,3-butadiene (BD) in mice. METHODS:Male Kunming mice were fed with corresponding diets for 3 weeks to establish FA-C (normal folic acid group),FA-D (folic acid deficiency group) and FA-S (folic acid supplementation group) animal models. Then,each group of FA-treated mice was randomly divided into control,and BD low-and high-dose groups (0,13.82,1 382.14 mg/m³) and treated with the corresponding dosages of BD by inhalation for 2 weeks (6 h/d,5 d/week). Enzyme-linked immunosorbent assay (ELISA) was applied to detect the concentrations of FA in serum and genomic DNA methylation levels in liver tissue. The expression of methyltransferase DNMT1 and DNMT3a was quantified by qPCR and chromosomal damage in peripheral blood lymphocytes was detected by cytokinesis-block micronucleus test (CBMNT). RESULTS:After 3 weeks of FA treatment,the concentrations of serum folic acid were significantly lower in FA-D group but higher in FA-S group compared with the FA-C group. With increase of the BD doses,methylation of global DNA and expression of methyltransferase were decreased while the frequencies of micronuclei,nucleoplasmic bridge and nuclear bud were increased within the same FA treated group. On the other hand,under the same BD exposure conditions,FA-D group showed a decreased level of DNA methylation and methyltransferase expression and increased chromosomal damage while FA-S group resulted in an increased level of DNA methylation and mRNA level of methyltransferase 1,3a expression and decreased micronucleus rate of chromosome compared with FA-C group,especially in the high dose BD exposure conditions (P < 0.05 or 0.01). CONCLUSION:BD can induce global DNA demethylation and chromosomal damage in mice. Folic acid deficiency can aggravate the BD-induced abnormalities. However,folic acid supplementation has a protective effect against the BD-induced abnormalities.

OBJECTIVE:To establishment a zebrafish model with CCM3 gene knockdown and to investigate its usefulness as a biological monitoring model for low concentrations of cardiovascular toxicants in water. METHODS:The transgenic zebrafish line (VEGFR2:GFP) which expresses green flurescence proteins in blood vessels were used. The one-cell stage zebrafish eggs were injected with morpholino oligos (MO) at indicated doses. Laser scanning confocal microscopy was used to observe the intracranial vessels of zebrafish at 72 h. The zebrafish model with CCM3 gene knockdown was established. Karber's method was used to calculate the median lethal dose (LD₅₀) of lead for zebrafish. The wild-type and CCM3 gene knockdown zebrafish were exposed to low concentrations of lead (10 mg/L). Development of zebrafish of each group was documented. RESULTS:The LD₅₀ value for wild-type zebrafish was 31.24 mg/L. Zebrafish with CCM3 gene knockdownt were more susceptible to low concentrations of lead-exposure (10 mg/L) than the wild-type by displaying abnormal phenotype,such as decreased body length,pericardial edema,hemorrhage,egg yolk edema, spine distortions. Up to 72 h,after treatment with the low dose,the wild-type zebrafish did not show any abnormal phenotype. CONCLUSION:The CCM3 gene knockdown zebrafish can possibly be used as a sensitive biological monitoring model for cardiovascular toxicants in water.

OBJECTIVE:To investigate the use of siRNA to silence the breast cancer resistance protein (BCRP/ABCG2) on overcoming multidrug resistance in breast cancer cells. METHODS:The morphologic,particle size,surface potential of magnetic nanoparticles (polyMAG) were characterized by transmission electron microscopy and by zeta potential analyzer,and the binding capacity of siRNA and polyMAG were investigated by gel retardation assay. We used polyMAG as a carrier to deliver several concentrations of siRNA (5,10,20,50,100 nmol/L) effectively into MCF-7/ADR cells. Quantitative real-time PCR (qPCR) and Western blotting were used to detect ABCG2 levels. CCK-8 and Calcein-AM/PI were stained and used to detect cell proliferation. RESULTS:The average size and zeta potentials of polyMAG was about 100 nm and 29.6 mv. Gel retardation assay confirmed that siRNA was well packaged into polyMAG under polyMAG/siRNA ratios 1:1 and 1:2. Cell viability of MCF-7/ADR was reduced while siRNA concentrations changed from 20 nmmol/L to 100 nmmol/L,compared with non-siRNA transfection (P < 0.05). Expression of ABCG2 mRNA was significantly down-regulated in the 20 nmol/L polyMAG-siRNA transfected group,which was about 1/8 of the un-transfected group (P < 0.05),and apoptotic cells were significantly increased compared with other groups. 50 nmol/L polyMAG-siRNA significantly interfered with the expression of ABCG2 protein (P < 0.05). CONCLUSION:polyMAG-siRNA effectively perturbed the ABCG2 expression and enhanced DOX susceptibility of MCF-7/ADR to overcome multidrug resistance in breast cancer cells.

OBJECTIVE:To investigate the involvement of mTOR-regulated SNCG expression in the control of radiation sensitivity in T47D breast cancer cells. METHODS:Expression levels of tumor-related genes in T47D cells were investigated after their irradiation with different doses of γ-rays. Western blot analysis was performed to determine the level of SNCG expression with or without the treatments of phosphatidic acid (PA;an mTOR pathway activator),and with 4 Gy γ-rays with or without PA treatment. Furthermore,SNCG expression was inhibited by transfection with SNCG-siRNA,and colony formation assay was performed to document radiation sensitivity. RESULTS:After γ-rays irradiation,there was a remarkable up-regulation of mTOR expression in T47D cells. However,radiation sensitivity was altered in PA-treated cells,suggesting that mTOR pathway was involved in the regulation of radio-sensitivity in these cells. From the qPCR and Western blot analyses,SNCG expression was up-regulated in either γ-rays irradiated or PA-treated T47D cells,indicating that mTOR-mediated the expression of SNCG which reduced the radiation sensitivity in T47D cells. Furthermore,transfection of cells with SNCG-siRNA significantly suppressed colony formation,suggesting that knockdown of SNCG expression can improve radiation sensitivity in breast cancer cells. CONCLUSION:Our study show that mTOR mediated the expression of SNCG and the subsequent radiation sensitivity of breast cancer cells. Our data suggest that down-regulation of SNCG expression may increase the sensitivity of radiotherapy and this may be helpful for the treatment of the patients with breast cancer.

OBJECTIVE:To study the effect of di(2-ethylhexyl) phthalate (DEHP) on metabolism in livers of female rats. METHODS:64 female Wistar rats were randomly divided into four groups:control (corn oil) and 150,300,600 mg/mL DEHP groups with 16 rats per group. Different doses of DEHP were administered via gavage,daily in volume of 2.5 μL/g,for 6 months. At 3 and 6 months,rat urine samples were collected. Rats were sacrificed at 6 months later. The liver,spleen and kidney were removed,and the ratio of viscera was calculated. Liver tissue sections were prepared and their morphological changes were observed by HE staining. The contents of DEHP and its metabolite,mono-ethylhexyl phthalate (MEHP),in urine were measured by high performance liquid chromatography (HPLC) at 2 time points. The content of MEHP in DEHP and its metabolite,mono-ethylhexyl phthalate,was measured by high performance liquid chromatography (HPLC). RESULTS:Compared with the control group,there was no significant difference in the general growth of the rats in each dose group,but the ratio of the dirty body in the liver increased. HE staining showed slight changes in the liver tissues of the 300 and 600 mg/mL DEHP rats. At 3 months,the levels of DEHP and MEHP in the urine of the 300 and 600 mg/mL DEHP groups were significantly increased with the increase of DEHP dose,the difference was statistically significant (P < 0.05). At 6 months,the levels of DEHP and MEHP in the urine of the 600 mg/mL DEHP group were higher than those of the other three groups (P < 0.05). CONCLUSION:DEHP in DEHP and DEHP and MEHP in the female rats were increased which exceeded the normal metabolic capacity,therefore,the increase might have caused some damage to the liver.

OBJECTIVE:To explore the effect of DDX46 knockdown by RNA interference on proliferation and apoptosis in esophageal squamous cell carcinoma Eca-109 cells. METHODS:DDX46 mRNA expression in Eca-109 and normal esophageal epithelium cell Het-1A were quantified using quantitative real-time PCR (qPCR). Then,DDX46 -shRNA-mediated RNA interference was applied to silence DDX46 in Eca-109. Subsequently,cell viability was measured by MTT assay,cell colony-forming capacity was measured by colony formation assay,cell cycle progression and apoptosis were determined by flow cytometry. Western blot analysis was used to detect changes of apoptosis signaling molecules. RESULTS:DDX46 mRNA was significantly up-regulated in Eca-109 cells compared with that in Het-1A cells (P < 0.01). DDX46 knockdown in Eca-109 led to reduced cell proliferation (P < 0.01),reduced number and size of cell colonies (P < 0.01),increased percentage of G1-phase cells,decreased percentage of S-phase cells (P < 0.05),and increased percentage of apoptotic cells (P < 0.01). Western blot analysis showed that the knockdown effectively suppressed the expression of DDX46 and up-regulated the expression of cleaved Caspase-3 and cleaved PARP-1 (P < 0.01). CONCLUSION:DDX46 mRNA was over-expressed in Eca-109 cells. Targeted silencing of the DDX46 gene inhibited cell proliferation,arrested cell cycle in the G0/G1 phase,and induced cell apoptosis. The DDX46 silencing effect could be mediated by positive regulation of the apoptosis signaling pathway which played the roles of inhibiting Eca-109 cells growth and inducing apoptosis.

OBJECTIVE:To study the correlation between telomere length of peripheral blood leukocytes and noise-induced hearing loss. METHODS:In a factory that produced computer shells,all posts that had noise intensity L_{EX,8 h} ≥ 85 dB(A),as determined by SV104IS wireless noise intensity detector,were selected. Questionnaires were used for collecting general and occupational information from workers and real-time polymerase chain reaction was used to detect telomere length of their peripheral blood leukocytes. Ordinal ploytomous logistic regression was used for analyzing the correlation between telomere length and noise-induced hearing loss. RESULTS:The average noise intensity among L_{EX,8 h} of overproof posts in the factory was (91.30±3.22) dB(A). This study recruited 127 noise-exposed workers consisting of 43 normal hearing workers (33.86%),75 abnormal high-frequency hearing loss workers (59.06%),9 abnormal speech-frequency hearing loss accompanied with abnormal high-frequency hearing loss (7.09%). Statistical significance was found in relative telomere length between different levels of noise-induced hearing loss (P < 0.05). It was revealed that noise intensity,age,gender and relative telomere length were factors related to noise-induced hearing loss (P < 0.05). CONCLUSION:Relative telomere length of peripheral blood leukocytes can potentially be used as a biomarker of susceptibility for noise-induced hearing loss.

OBJECTIVE:To determine genotype-specific distribution of human papillomavirus (HPV) in women and to identify its relationship with cervical carcinoma in Chaoshan population. METHODS:Data from 30 000 women who were subjected to cervical HPV DNA testing were retrospectively evaluated in a hospital-based study. A total of 8 739 women,who had both HPV typing and pathological examination,were investigated. RESULTS:Overall,30.00% (9 000/30 000) of women were HPV positive,the frequency of HPV infection was 58.29% (5 094/8 739) in women with cervical cytological abnormalities,and the infection rate for patients with chronic cervicitis was 40.00% (1 088/2 720),patients with CIN I was 53.59%(1 396/2 605),patients with CIN was Ⅱ/Ⅲ 71.61% (1 647/2 300) and patients with cervical carcinoma was 86.45%(963/1 114). HPV 16 (14.02%) was the most common type for all cervical cytological abnormalities,followed by 18 type 6.28%(549),33 type 5.23%(457),58 type 4.46%(390) and 52 type 3.71% (324),while the more common HPV 16 type in cervical carcinoma with the highest infection (49.92%) was in women aged over 60. CONCLUSION:Our data indicate a relatively high prevalence of HPV 16,52,33,58 and 18 in women,their positive association with cervical cytological abnormalities. Specifically,the HPV 16 was the most common subtype among senior women with cervical carcinoma.

OBJECTIVE:To compare the presence of chemical elements in plasma between gastric cancer patients and healthy people. METHODS:Using a case-control design,we focused on people who had lived in Xianyou for 10 years or more. Among them,we identified newly discovered cases of pathologically confirmed gastric cancer (299 cases) and 295 health cases. Among these cases,we obtained general situation information,living habits,etc. using questionnaires and collected blood samples. Blood samples were subjected to microwave digestion and chemical elements in plasma were analyzed by inductively coupled plasma mass spectrometry. The presence of chemical elements between the two groups were compared using non-parametric tests. RESULTS:Cu,Mg,and Cu/Zn in the case group were significantly higher than that in the control (P < 0.01). However,Mo,Ca in the case group were significantly lower (P < 0.01). Multivariate analysis showed that Cu (OR=2.71),Mg (OR=2.26),Cu/Zn (OR=3.88) were significantly associated with increased risk for gastric cancer but Mo (OR=0.48),Ca (OR=0.17) with significantly reduced risk. CONCLUSION:Levels of some plasma chemical elements in gastric cancer patients were significantly different from that of the normal population.

OBJECTIVE:To provide positive reference for teratogenicity test on food,health care products and cosmetics,we explored teratogenic effects of three different positive control materials. METHODS:According to the gestation time of SD rats,pregnant rats were randomly divided into control,aspirin (280 mg/kg),Bis-A-TDA (1 mg/kg) and CP (12 mg/kg) groups. Their body weights on days 0,6,9,12,15 and 20 of pregnancy were recorded. On day 6-15 of gestation,the aspirin and Bis-A-TDA groups were administrated corresponding solutions,while CP group were injected ip with CP on day 12. Pregnant rats were inspected by cesarean section and weights of uterus,uterine and fetuses were recorded. The number of corpus luteum,implantation point,and embryonic development were examined. Appearance of fetus was examined,and fetal weight,body length and tail length of each fetus were measured. 1/2 of the live fetus were stained for skeletal examination and the others were fixed for visceral examination. RESULTS:In the aspirin group,body weight and weight increment of the pregnant rats,the weight of the uterus and uterine and fetal were reduced. The rates of live fetus,fetal weight,the length of body and tail were decreased significantly (P < 0.05,P < 0.01). In the Bis-A-TDA group,the rates of live fetus,fetal weight,the length of body and tail were significantly decreased (P < 0.01). In the CP group,weight increments of the pregnant rats,and the weight of the uterus and uterine and fetal,and the rate of live fetus were decreased significantly (P < 0.05,P < 0.01). The teratogenic rate of appearance, skeleton and viscera of three different positive control materials were statistically significant (P < 0.01). There were more teratogenic types and higher teratogenic rates in the Bis-A-TDA and CP groups. CONCLUSION:Under the conditions of our experiment,Bis-A-TDA (1 mg/kg) and CP (12 mg/kg) induced more teratogenic types,and higher teratogenic rate,therefore they were more sensitive positive control materials on teratogenicity test.