Objective To investigate the effects of immune system and immuno-related miRNAs expression in low-dose benzene and its homologue mixed-exposed workers in a short time. Methods A total of 40 workers were recruited from a petrochemical factory ,and their blood samples were collected before and after work to detect the levels of IgA,IgG,IgM,IFN-γ,IL-1β,IL-2,IL-8,TNF-αin serum and miRNA-146a,-155 expression in the peripheral blood mononuclear cell. In the meanwhile ,we measured the individual benzene and its homologue exposure level of recruited workers. Results According to the concentration of benzene ,the subjects were divided into benzene-exposed group(20 workers)and control group(20 workers). And toluene exposure level in benzene-exposed group was significantly higher than that in control group(P<0.05). Significant decrease of IgG was found in benzene-exposed workers after one work shift,compared with control group(P<0.05). Moreover,the interaction between benzene and toluene was significant correlated with the decrease of IL-1β and TNF-α(P < 0.05). Additionally,the interaction between miRNA-155 and miRNA-146a was significant correlated with the decrease of TNF-α(P<0.05). Conclusions:Occupational exposure to low-dose benzene and toluene in a short time could decrease immune function. And there may be an effect of miRNAs in the regulation of cytokine.

Objective: To investigate the value of DNA content in comet tail (TailDNA) in predicting the changes in peripheral blood cell counts in workers exposed to benzene. Methods: In 2011, cluster sampling was used to select 150 male workers exposed to benzene in a petrochemical factory. Cubital venous blood and urine samples were collected for routine blood rest, comet assay, and measurement of s-phenylmercapturic acid (SPMA) and urine creatinine. The population was divided into groups according to urinary SPMA or TailDNA, and routine blood test results within 3 years were collected to analyze the changes in blood cell counts. Results: The low-SPMA group had significantly higher white blood cell and neutrophilcounts in all years than the high-SPMA group (P<0.05) . The low-Tail DNA group had a significant increase in platelet count from 2012 to 2014 (P<0.05) , while the high-Tail DNA group had no significant change (P>0.05) . During the 4-year period, the high-TailDNA group had a significantly lower red blood cell count than the low-TailDNA group (P<0.05) . The high-TailDNA group showed a gradual reduction in white blood cell count over time (β=-0.113, P<0.05) , and the low-TailDNA group showed no trend of the reduction in white blood cell count (P>0.05) . Conclusion TailDNA can be used to predict the changes in blood cell counts in workers exposed to benzene.

Objective To investigate the expression of apolipoprotein (Apo) F in hepatocellular carcinoma (HCC) and its application value in the prognosis of patients with HCC. Methods 50 HCC samples were procured from patients undergoing surgical resection in the Third Affiliated Hospital of Sun Yat-sen University between September 2015 and September 2016, and all the samples were confirmed by postoperative pathological examination. The informed consents of all patients were obtained and the local ethical committee approval was received. There were 37 males and 13 females, aged from 31-67 with a median age of 53 years old. The expression of ApoF mRNA in HCC tissues was detected by RT-PCR. The expression profile was analyzed by using data from the Gene Expression Omnibus (GEO). The expression of ApoF between two groups were compared by t test. Correlation analysis of clinical related parameter was conducted by Chi-square test, and survival prognosis was analyzed by Kaplan-Meier test and Log rank test. Results The average relative expression of ApoF mRNA in HCC tissues was 0.15±0.07, significantly lower than 0.55±0.09 in the adjacent tissues (t=-6.26, P<0.05). GEO online analysis showed that expression of ApoF was significantly correlated with the status of liver cirrhosis, and most HCC patients with liver cirrhosis presented low expression of ApoF (χ2=4.626, P<0.05). The 5-year disease-free survival was respectively 55.9% and 32.0% in ApoF high expression group and low expression group, where significant difference was observed (χ2=3.939, P<0.05). Conclusions Low expression of ApoF exists in HCC tissues, and it is related to the liver cirrhosis status of patients. Patients with low ApoF expression present poorer prognosis. ApoF plays a role in inhibiting the cancer.

Objective To explore the impact of telomerase regulation factor PinX1 to the proliferation and invasion ability of hepatoma cells. Methods Hepatoma cells PinX1-7721 (experimental group) with stable expression of PinX1 as well as control cell VECTOR-7721 (control group) were constructed. The expression of PinX1 mRNA was detected by RT-PCR. The proliferation ability and clonality of hepatoma cells were detected by CCK-8 method and plate clonality assay, and the invasion ability of hepatoma cells by Transwell assay. Comparison of the experiment data was conducted by t test. Results Expression level of PinX1 mRNA in experiment group was (13.9±2.0)×10-3, which was significantly higher than (1.1±0.2)×10-3in control group (t=10.98, P<0.05). A450of the cells on 1-7 d in experiment group was respectively 0.260±0.004, 0.340±0.008, 0.450±0.040, 0.500±0.020, 0.730±0.030, 1.350±0.040 and 1.640±0.050, which were significantly lower than 0.280±0.009, 0.410±0.007, 0.680±0.044, 0.730±0.029, 0.850±0.070, 1.700±0.020 and 2.080±0.280 in control group (t=-5.82, -12.99, -6.36, -5.96, -28.42,-18.98, -5.08; P<0.05). The plate clonality assay results showed that the clone formation quantity of cells in experiment group was 143±32, which was significantly lower than 305±25 in control group (t=-6.91, P<0.05).Transwell assay results showed that the quantity of trans-membrane cell in experiment group was 230±16, which was significantly lower than 650±30 in control group (t=-21.40, P<0.05). Conclusion PinX1 could inhibit the proliferation and invasion ability of hepatoma cells.

Objective To investigate expression of plasmalemmal vesicle-associated protein (PLVAP) in hepatocellular carcinoma (HCC) tissues and its relationship with clinicopathological features.Methods Tissue specimens were collected from 108 patients with HCC in the Third Affiliated Hospital of Sun Yat-sen University from January 2013 to December 2015.92 patients were male and 16 female,aged (48±5) years on average.The informed consents of all patients were obtained and the local ethical committee approval was received.The expression level of PLVAP was analyzed based on the data of HCC in public databases.The expression level of PLVAP mRNA in HCC and paracarcinoma tissues was detected by RT-PCR,and the relationship between the expression of PLVAP and clinicopathological characteristics of HCC was analyzed.The relationship between PLVAP and prognosis of HCC patients was investigated with the data from cancer genome atlas (TCGA) database.The expression levels of PLVAP mRNA between HCC tissues and para-carcinoma tissues were compared by Kruskal-Wallis rank-sum test.Correlation analysis was performed by Chi-square test.Survival analysis was conducted by Kaplan-Meier survival curve and Log-rank test.Results According to Human Protein Atlas and Oncomine databases,the expression level of PLVAP in HCC tissues was significantly higher than that in normal liver tissues.RT-PCR showed that the median expression level of PLVAP mRNA in HCC tissues was 0.172(0.004-0.607),significantly higher compared with 0.091(0.002-0.513) in para-carcinoma tissues (Z=6.839,P＜0.05).The expression level of PLVAP in HCC patients was significantly correlated with TB,tumor size and microvascular invasion (x2=4.183,3.924,6.075;P＜0.05).In PLVAP high expression group,the overall survival and tumor-free survival were 58.8(0.5-107.0) and 42.2(0.1-67.2) months,where no significant difference from 55.7(0.2-120.7) and 20.9(0.1-109.4) months in PLVAP low expression group (x2=0.054,0.065;P＞0.05).Conclusions The expression level of PLVAP is significantly correlated with the development and progression of HCC,whereas it is probably not associated with the prognosis of HCC patients.

OBJECTIVE: To investigate the effects of subacute systemic inhalation exposure of 1,2-dichloroethane(1,2-DCE) on learning and memory in NIH mice. METHODS: Forty-five specific pathogen free healthy 7-week-old NIH mice were randomly divided into control,low-dose and high-dose groups with 5 female mice and 10 male mice in each group. The mice were exposed to 1,2-DCE at dosages of 0. 00,100. 00 and 350. 00 mg/m3 for 6 hours per day for consecutive 28 days by dynamic systemic inhalation. The neurobehavioral tests of mice were performed before and after the first to fourth weeks of exposure using the Morris water maze test. RESULTS: There was no significant difference in body weight and swimming speed among the three groups of mice( P > 0. 05). The navigation experiment results showed that the escape latency of mice in both low-and high-dose groups were longer than that of the control group at the same time point(P < 0. 05) during 1-4 weeks after exposure. In the control group,the escape latency was shorter than that of the same group before exposure( P < 0. 05). The escape latency of high-dose group prolonged with the increase of exposure time,and in the 4 th week the escape latency was significantly higher than that of the same group before exposure( P < 0. 05).The experiment results of space exploration indicated that the first time of crossing platform in low-and high-dose groups were longer than that of the control group at the second to the fourth week( P < 0. 05). The target quadrant retention time and the number of crossing the platform in the low-and high-dose groups were lower than those in the control group( P <0. 05). CONCLUSION: Subacute inhalation exposure of 1,2-DCE can impair the learning and memory ability of NIH mice.The high-dose exposure may reduce learning ability in mice in a time-effect manner.

OBJECTIVE: To study the effect of aquaporin 4(AQP4) in regulating the permeability of blood-brain barrier(BBB) induced by subacute 1,2-dichloroethane(1,2-DCE) inhalation. METHODS: Specific pathogen free healthy CD-1 male Aqp4 genetically engineered mice(Aqp4~(+/+)and Aqp4~(-/-)) were randomly divided into control and low-, medium-and high-dose groups. The mice were exposed to 1,2-DCE at the dosages of 0.00, 100.00, 350.00 and 700.00 mg/m~3 for 6 hours per day for consecutive 28 days by systemic dynamic inhalation. After the end of 1,2-DCE exposure, the BBB permeability was evaluated by Evans blue staining. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of genes related to BBB tight junction protein(Tjp)1, Tjp2, Tjp3, claudin(Cldn)3, Cldn5, Cldn11, occludin(Ocln), matrix metalloproteinase(Mmp)2, Mmp9 and Na-K-Cl cotransporter-1(Nkcc1). RESULTS: The BBB permeability in mice showed significant change with 1,2-DCE dose and Aqp4 genotype(P<0.01). The BBB permeability of Aqp4~(+/+) genotype mice was higher in low-, medium-and high-dose groups than that of control group(all P values were <0.05). The permeability of BBB was lower in Aqp4~(+/+) genotype mice in the control group than that of Aqp4~(-/-) genotype mice in the same group(P<0.05), but BBB permeability was higher in Aqp4~(+/+) genotype mice in the three dose groups than that of Aqp4~(-/-) genotype mice in the same group(all P values were <0.05). The Cldn3 and Olcn mRNA relative expression in the brain cortex had statistical difference in mice with different genotype(all P values were <0.01). The mRNA relative expressions of Cldn3 and Olcn in the brain cortex were higher in Aqp4~(-/-) genotype mice than that of Aqp4~(+/+) genotype mice(all P values were <0.01). The relative mRNA expression levels of Tjp1, Tjp2, Tjp3, Cldn5, Cldn11, Mmp2, Mmp9 and Nkcc1 in the cerebral cortex of mice were not statistically significant in aspect of 1,2-DCE exposure dose and genotype(all P values were >0.05). CONCLUSION: Exposure to 1,2-DCE can increase BBB permeability in mice, and the mechanism may be associated with 1,2-DCE-induced down-regulation of Aqp4 and up-regulation of mRNA expression of the cerebral cortex TJP-related molecules Cldn3 and Ocln.

OBJECTIVE: To explore the molecular mechanism underlying 1,2-dichloroethane(1,2-DCE) induced apoptosis by screening differentially expressed proteins in human astrocytes( HAs). METHODS: HAs were cultured in complete medium with 1,2-DCE at various concentrations of 0-80 or 0-40 mmol/L. After 24 hours,apoptosis of HAs was evaluated using flow cytometry and staining with annexin Ⅴ-fluoresce in isothiocyanate and propidium iodide. An AAH-APO-1-2 protein chip was used to screen differentially expressed proteins and quantitative real-time polymease chain reaction(qRT-PCR) was used to verify related differentially expressed genes(DEGs). RESULTS: At 1,2-DCE concentrations of0-80 mmol/L,the total apoptosis rate of HAs increased with 1,2-DCE concentrations in a dose-dependent manner( P <0. 01). Seven different kinds of proteins were screened out by apoptotic protein chip. Among them,the expression of insulin-like growth factor-binding protein( IGFBP)-1,IGFBP-4 and cytochrome C( Cyto C) were up-regulated,while the expression of P27,cysteine aspartic acid specific protease-3( Caspase-3),B-cell lymphoma-2 interacting mediator of cell death( BIM) and BH3 interacting domain death agonist( BID) were down-regulated compared with the control group. The result of DEGs verified by qRT-PCR showed that the expression of mRNA of IGFBP-1,IGFBP-4 and Cyto C at 1,2-DCE concentrations of 40 mmol/L was up-regulated. This result was in consistent with the trend of target expression in the protein chip. The mRNA expression of Caspase-3,BIM and BID was also up-regulated. CONCLUSION: 1,2-DCE induces apoptosis of HAs through mitochondrial pathway.

OBJECTIVE: To predict the sensitizing potency and optimal sensitization dose of trichloroethylene( TCE) by an in vitro skin sensitization test on a human acute monocytosis cell line( THP-1).METHODS: THP-1 cells were cultured in vitro and exposed to 2,4-dinitrochlorobenzene( DNCB),sodium dodecyl sulfate( SDS),tert-butylhydroquinone( tBHQ)and TCE for 24 hours.Flow cytometry was used to detect the expression of cell surface marker such as cluster of differentiation( CD) 86 and CD54,and the optimal dose range for sensitization detection was determined.With the relative fluorescence intensity( RFI),CD86 ≥ 150 and CD54 ≥ 200 as the standard,the sensitizing potency and optimal sensitization dose of TCE were predicted.RESULTS: The concentration range of reagents for sensitization test on THP-1 cells was the dose range at which the relative cell survival rate reached 75.0%-100.0%.DNCB at the doses of 20.83,25.00 and 30.00 μmol/L,tBHQ at the dose of 5.80 μmol/L,TCE at the doses of 8.33,10.00 and 12.00 mmol/L,can cause sensitivity.SDS was recognized as a negative sensitizer.The expression of CD86 and CD54 was the highest when the concentration of TCE was 8.33 mmol/L,which was considered as the best sensitization dose.CONCLUSION: The optimum sensitization dose of TCE is 8.33 mmol/L,which can provide the basis for dose design in future study of TCE sensitization pathways.

OBJECTIVE: To investigate the effect of 1,2-dichloroethane(1,2-DCE) acute inhalation exposure on the differential gene expression of phase Ⅰ metabolic enzymes. METHODS: The specific pathogen free SD rats were randomly divided into control group(16 rats), low-and high-dose groups(24 rats in each group, half males and half females). Low-and high-dose group were given daily 600, 1 800 mg/m~(3 ) of 1,2-DCE, and the control group given the fresh air by dynamic inhalation for 8 hours per day for consecutive 7 days. After the end of exposure, the relative mRNA expression of cytochrome P450 2 E1(CYP2 E1), alcohol dehydrogenase(ADH1) and acetaldehyde dehydrogenase 3 alpha 1(ALDH3α1) in the liver tissue was detected by real-time fluorescence quantitative polymerase chain reaction. RESULTS: The relative expression of CYP2 E1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ADH1 in male low-and high-dose groups was higher than that in male control group(P<0.05). The relative expression of ADH1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ALDH3α1 in high-dose group was higher than that in control group and low-dose group(P<0.05). CONCLUSION: High dose 1,2-DCE could increase the gene expression of phase Ⅰ metabolic enzymes in rat liver. The 1,2-DCE has more obvious effect in male rats than in female rats.