Objective To investigate the clinical characteristics and influencing factors of moderate-to-severe pulmonary ventilation dysfunction in occupational pneumoconiosis (hereinafter referred to as "pneumoconiosis") patients complicated with pulmonary tuberculosis. Methods A total of 136 male pneumoconiosis patients complicated with pulmonary tuberculosis suffering different degrees of pulmonary ventilation dysfunction were selected as the study subjects using the judgmental sampling method. Patients were divided into mild dysfunction and moderate-to-severe dysfunction groups based on the degrees of pulmonary ventilation dysfunction. Clinical data from patients of these two groups were collected, and influencing factors of pulmonary ventilation dysfunction were analyzed. Results The prevalence of mild dysfunction and moderate-to-severe dysfunction among the study subjects was 39.0% and 61.0%, respectively. The proportion of patients with moderate-to-severe pulmonary ventilation dysfunction increased with the progression of pneumoconiosis (P<0.05). Patients in moderate-to-severe dysfunction group had higher rates of dyspnea, elevated C-reactive protein, coexisting chronic obstructive pulmonary disease (COPD), and a history of lung infections within the past two years compared with those in the mild dysfunction group (all P<0.05). The result of multivariate logistic regression analysis showed that the degree of pneumoconiosis, complicated with COPD, and a history of lung infections within the past two years were risk factors for moderate-to-severe pulmonary ventilation dysfunction (all P<0.05). Specifically, higher degree of pneumoconiosis was associated with a greater proportion of moderate-to-severe dysfunction, and patients complicated with COPD or had a history of lung infections within the past two years were more likely to experience severe pulmonary ventilation dysfunction. Conclusion The degree of pneumoconiosis, complicated with COPD, and a history of lung infections within the past two years are influencing factors of moderate-to-severe pulmonary ventilation dysfunction in patients with pneumoconiosis combined with pulmonary tuberculosis. Early detection of pneumoconiosis progression, timely diagnosis of COPD and lung infections, and appropriate treatment such as antifibrotic agents, inhaled bronchodilators, and anti-infective therapies are recommended.

Objective To study the relationship between chemotherapeutic resistance to 5-fluorouracil (5-FU) and Golgi phosphoprotein 3 (GOLPH3) expression in human colon cancer cell line HT29.Methods HT29 cells were divided into four groups:control group,siRNA transfection group,experimental group 1 (30 μmol/L 5-FU),and experimental group 2 (siRNA-GOLPH3 + 30 μmol/L 5-FU).The silencing effect of GOLPH3 gene was detected by RT-PCR and Western blot.The tumor proliferation and formation ability of HT29 cell were measured respectively by MTT and plate colony formation assay.The protein expressions of GOLPH3,P-gp and β-catenin in HT29 cell were detected by Western blot.Results Compared with the control group,the expression level of GOLPH3 mRNA (1.000 ± 0.078 vs.0.147 ± 0.021,t =12.296,P ＜ 0.01) and protein(1.003 ± 0.235 vs.0.077 ± 0.399,t =6.723,P ＜ 0.01),the absorbance(A value) (1.000 ± 0.082 vs.0.769 ± 0.086,t =3.885,P ＜ 0.01) and the tumorigenesis (430 ±36 vs.297 ± 21,t =5.492,P ＜ 0.01) in transfection group significantly reduced.The A value (0.803 ± 0.101 vs.1.050 ± 0.140,t =2.855,P ＜ 0.05;0.242 ± 0.091 vs.1.050 ± 0.140,t =9.664,P ＜ 0.001) and the tumorigenesis (73 ± 8 vs.427 ± 29,t =20.363,P ＜ 0.001;305 ± 22 vs.427 ± 29,t =5.840,P ＜ 0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group,while the A value (t =8.264,P ＜ 0.001) and the tumorigenesis (t =17.346,P ＜ 0.001) of experimental group 2 significantly decreased than that experimental group 1.Compared to the control group (0.967 ± 0.094,1.001 ± 0.199,1.000 ± 0.101),the expressions of GOLPH3,P-gp and β-catenin in experimental group 1 (3.634 ± 0.574,2.424 ± 0.261,2.324 ± 0.418) significantly upregulated (t =7.944,P ＜ 0.01;t =7.520,P ＜ 0.01;t =5.330,P ＜ 0.01),while the expressions of these proteins in experimental group 2(0.520 ±0.176,0.466 ±0.159,0.933 ±0.049) were significantly lower than that in experimental group 1 (t =8.983,P ＜ 0.01;t =11.106,P ＜ 0.001;t =5.724,P ＜ 0.01).Conclusion Expression of GOLPH3 is associated with resistance to 5-FU in colon cancer HT29 cell by activating Wnt/β-catenin signal pathway.

OBJECTIVE: To explore the effects of nano-silicon dioxide( SiO_2) on the survival and poly( ADP-ribose)polymerase-1( PARP-1) expression in human bronchial epithelial cells( 16 HBE cells). METHODS: i) The 16 HBE cells were treated with nano-SiO_2 at concentrations ranging from 0 to 100 mg/L for 24. 0 hours,and CCK-8 assay was used to examine cell viability. ii) The 16 HBE cells were divided into 6 groups: solvent control group( equal volume solvent treatment),micro-SiO_2 control group( treated with 20 mg/L micro-SiO_2),5,10,and 20 mg/L nano-SiO_2 groups( treated with the corresponding final dose of nano-SiO_2),and curcumin group. The curcumin group was given pretreatment with curcumin at a final concentration of 10 μmol/L for 2. 0 hours followed by treatment with a final concentration of 20 mg/L of nano-SiO_2. Cells in each group were harvested at time points of 4. 0,12. 0 and 24. 0 hours after treatment. The relative expression of PARP-1 mRNA and protein in 16 HBE cells was detected by quantitative real-time polymerase chain reaction and Western blotting respectively. RESULTS: i) The survival of 16 HBE cells decreased with increasing nano-SiO_2 treatment dose,showing a dose-effect relationship( P < 0. 01). ii) The expression of PARP-1 mRNA and protein in 16 HBE cells were dose-dependently decreased after nano-SiO_2 stimulation at the 12. 0 and 24. 0 hours time points( P < 0. 01). The expression of PARP-1 mRNA and protein in 5,10,and 20 mg/L nano-SiO_2 groups decreased at the above mentioned time points( P < 0. 05),compared with the solvent control group at the same time points. The expression of PARP-1 mRNA and protein in 20 mg/L nano-SiO_2 group was lower than that in the micro-SiO_2 control group at the same 12. 0 and 24. 0 hours time point( P < 0. 05). The above two indexes of cells were higher in curcumin group than that of 20 mg/L nano-SiO_2 group at the 12. 0 hours time point( P < 0. 05). CONCLUSION: Nano-SiO_2 stimulation can lead to decrease survival of 16 HBE cells in a dose-dependent manner and down-regulation of PARP-1 expression may be one of the mechanisms of proliferation and inhibition of 16 HBE cells induced by nano-SiO_2. Curcumin has certain protective effect on nano-SiO_2-induced 16 HBE cell injury.

Objective: To detect the alterations of telomerase activity and the expression for oxidative stress responsive genes related with senescence during cellular replicative senescence and hydrogen peroxide-induced premature senescence in human embryonic lung fibroblasts (HELFs) in vitro. Methods: The HELFs were divided into young cells (22 population doubling levels, 22PDL) , mid-aged cells (35PDL) and replicative senes-cent cells (49PDL) and premature senescent cells induced by H₂O₂(premature senescence, PS). The telomerase activity was detected by ELISA assay during cellular replicative and premature senescence. The mRNA level of oxidative stress responsive genes related with senescence for Foxo1, Foxo3, Pdx1, apoA-I and MMP1 was per-formed by RT-Q-PCR separately. Results: The mRNA level for Foxo1, Foxo3, apoA-I and Pdx1 was decreased separately during cellular replicative senescence compared to that in the young-stage cells with statistical signifi-cance (P<0.05). The expression of MMP1 was up-regulated 5.1-fold obviously (P<0.05). In premature senes-cence, the mRNA level was only decreased for Foxo1, Foxo3 and apoA-I, but up-regulated 2.3-fold and 6.2-fold for Pdx1 and MMP1 respcetively vs 22PDL significantly (P<0.05). The telomerase activity in young cells was not detected, and it increased in mid-aged cells and replicative senescence stages during cellular replicative se-nescence as compared to 22PDL with statistical significance (P<0.05). The telomerase activity in premature se-nescence was highly active. Conclusion The expression for genes related with senescence has differences be-tween replicative and premature senescence and hydrogen peroxide modifies their expression levels. The telomer-ase activity has been going up with increased PDLs.

The controversial results of several studies suggest that certain everyday-use chemicals may be linked to breast cancer. ln recent years, extensive researches have been carried out to under-stand breast carcinogenesis and the hedgehog(Hh) signaling pathway has emerged as a critical determi-nant of human breast cancer. Aberrant Hh signaling in adults results in carcinogenesis, angiogenesis, and metastasis. This review is focused on the Hh signaling pathway and chemicals in the regulation of breast cancer development and provide an updated survey of pre-clinical and clinical trials of novel strategies to target them.

OBJECTIVE To explore the effect of bisphenol A (BPA) on the differentiation potential of embryonic stem cells, and provide an experimental basis for evaluation of safety of BPA. METHODS Mouse embryonic fibroblasts (MEFs) and embryonic stem cells (ESCs) were treated with BPA 0.1, 1, 10, 100 and 1000 μmol.L-1 for 8 d respectively. The viability of MEFs and ESCs was measured by CCK-8 and lC50 was calculated. The mRNA expression of α-myosin heavy chain in ESCs was tested by RT-PCR to determine lD50 . The embryonic body cultured by suspension method was treated with BPA 0.001, 0.01, 0.1 and 1 μmol.L-1 for 10 d respectively. The changes of marked genes in each blastoderm were detected by RT-PCR. RESULTS lC50 of BPA to mouse ESCs was 5.22×10-4 mol.L-1 , and to MEFs was 6. 25 × 10-4 mol.L-1 . lD50 of BPA to mouse ESCs differentiating to cardiomyocytes was 7.0×10-7 mol.L-1 . BPA 0.001 and 0.01 μmol.L-1 upregulated the expression of the marked genes of mesoderm, fetal liver kinase-1 and globin transcription factor 1. CONCLUSION BPA is a strong embry-otoxic compound. BPA of low concentration can promote the differentiation of mouse ESCs to mesoderm.

OBJECTIVETo study the effect of silicon dioxide nanoparticles on the expression and promoter region CpG islands methylation of (Poly [ADP-ribose] polymerase 1, PARP-1) gene in human HaCaT Cell.

METHODSHaCaT Cells were treated with nm-SiO₂at 0, 2.5, 5 and 10 µg/mL and micro-SiO₂at 10 µg/ml for 24 h and DAC treatment was given at 10 µg/ml group for 48 h. Real-time PCR and western blot assay was used to detect the expression of PARP-1 mRNA and protein. BSP (Bisulfite Pyrosequence, BSP) assay was used to detect the promoter region CpG islands methylation status of PARP-1 gene.

RESULTSAfter exposure to nano-SiO₂particles, compared to CTRL group, the mRNA and protein expression of PARP-1 in micro-SiO₂and 2.5 µg/ml group unchanged, but he mRNA and protein expression of PARP-1 in 5, 10 µg/ml as well as DAC group was down-regulated and there are statistical significance between CTRL group and 5, 10 µg/ml as well as DAC group and the PARP-1 promoter region CpG islands showed methylation.

CONCLUSIONnano-SiO₂can down-regulate PARP-1 expression in HaCaT Cell and this is associated with the change in the methylation of PARP-1 gene promoter region CpG islands induced by nano-SiO₂particles.

Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Humans ; Nanoparticles ; adverse effects ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Silicon Dioxide ; adverse effects

OBJECTIVETo put the insight into the trichloroethylene (TCE)-induced effect on the differential expression of subcellular proteins in human normal liver cell line (L-02).

METHODSThe membrane proteins and nuclear proteins of TCE-treated (8.0 mmol/L) group and controls were extracted by subcellular proteome extraction kit, respectively. The TCE-induced differentially expressions were analyzed by a two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization tandem time-of-flight spectrometry (MALDI-TOF-MS). Bioinformatics analysis was used to reveal the biological processes and predict transmembrane domains of differential expressed proteins. The expression of ATP synthase subunit beta (ATP5B), heterogeneous nuclear ribonucleoprotein H2 (hnRNP H2) and far up steam element-binding protein 1 (FUBP1) were measured under TCE treatment by Western blot.

RESULTSAfter TCE treatment for 24 h in L-02 cells, 14 membrane proteins and 18 nuclear proteins were identified as differential expression. After treated with TCE in concentrations of 0, 2.0, 4.0 and 8.0 mmol/L for 24 h, the relative levels of ATP5B expression were 1.00±0.03, 1.21±0.14, 1.25±0.12 and 1.48±0.17 (F = 8.51, P = 0.007), the relative levels of hnRNP H2 expression were 1.00±0.09, 1.22±0.15, 1.43±0.21, 1.53±0.17 (F = 6.57, P = 0.015), respectively; the relative levels of FUBP1 expression were 1.00±0.11, 0.91±0.07, 0.73±0.04 and 0.67±0.03 (F = 15.81, P = 0.001), respectively, which were consistent with the results in proteomics. The bioinformatics analysis showed that the most dominant biological process were involved in RNA processing (10 proteins, P = 2.46×10(-6)), especially in RNA splicing (9 proteins, P = 1.77×10(-7)).

CONCLUSIONThe exposure of TCE could alter the expression of membrane proteins and nuclear proteins in L-02 cells. These abnormal expressed proteins involved in RNA splicing would provide novel clues for further understanding of TCE-induced hepatotoxicity.

Blotting, Western ; Cell Line ; DNA Helicases ; DNA-Binding Proteins ; Hepatocytes ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H ; Humans ; Mitochondrial Proton-Translocating ATPases ; Proteome ; Proteomics ; RNA Processing, Post-Transcriptional ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trichloroethylene

OBJECTIVETo investigate the effect of poly-ADP-ribosylation in hexavalent chromium Cr(VI) induced cell damage.

METHODSThe study object, poly (ADP-ribose) glycohydrolase (PARG) deficient human bronchial epithelial cells (16HBE cells), was constructed previously by our research group. Normal 16HBE cells and PARG-deficient cells were treated with different doses of Cr (VI) for 24 h to compare the differences to Cr (VI) toxicity, meanwhile set up the solvent control group. On this basis, 5.0 µmol/L of Cr (VI) was selected as the exposure dose, after the exposure treatment, total proteins of both cells were extracted for two dimension fluorescence difference gel electrophoresis (2D-DIGE) separation, statistically significant differential protein spots were screened and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS), and further validated by Western blot.

RESULTSAfter Cr (VI) treatment, the survival rate of PARG-deficient cells was higher than normal 16HBE cells. When the doses reached up to 5.0 µmol/L, the survival rate of 16HBE cells and PARG-deficient cells were respectively (59.67 ± 6.43)% and (82.00 ± 6.25)%, the difference between which was significant (t = -4.32, P < 0.05). 18 protein spots were selected and successfully identified after 2D-DIGE comparison of differential proteins between normal 16HBE cells and PARG-deficient cells before and after exposure. The function of those proteins was involved in the maintenance of cell shape, energy metabolism, DNA damage repair and regulation of gene expression. The differential expression of cofilin-1 was successfully validated by Western blot. The expression level of cofilin-1 in the 16HBE cells increased after Cr (VI) exposure with the relative expression quantity of 1.41 ± 0.04 in treated group and 1.00 ± 0.01 in control group, the difference of which was statistically significant (t = -18.00, P < 0.05), while the expression level in PARG-deficient cells had no statistically significant difference (t = -8.61, P > 0.05).

CONCLUSIONMost of the identified differential proteins are closely related to tumorigenesis, suggesting that poly-ADP-ribosylation reaction may resist the cytotoxicity of Cr(VI) by inhibiting Cr (VI) induced tumorigenesis, which provides important reference data to clarify the mechanisms of poly-ADP-ribosylation in Cr (VI) induced cell damage.

Bronchi ; Cell Transformation, Neoplastic ; genetics ; Chromium ; Cofilin 1 ; DNA Repair ; Epithelial Cells ; Glycoside Hydrolases ; deficiency ; physiology ; Humans ; Tandem Mass Spectrometry

OBJECTIVETo reveal the role of poly-ADP-ribosylation and DNA methylation in carcinogenic process induced induced by Cr (VI), and to discuss the relations between them.

METHODSThe pre-established Poly (ADP-ribose) glycohydrolase (PARG) deficient cells and 16HBE cells were treated with different concentrations of Cr (VI), and the changes of total genomic DNA methylation level in different groups were detected by methylation immunofluorescent detection, as well as the changes of the activity of methyltransferases. Moreover, RT-PCR and western blotting method were applied to analyze the changes of expression of DNMT1, DNMT3a, DNMT3b and MBD2, upon the protein level.

RESULTSAfter treated by Cr(VI) for 24 h, the healthy 16HBE cells showed a significant lower level of genomic DNA methylation; however, there was no significant changes (P > 0.05) found in PARG deficient cells by immunofluorescence assay. When the dose of Cr (VI) reached 5.0 µmol/L, the activity of methyltransferases in 16HBE cells and PARG deficient cells (49.33 ± 2.65, 80.05 ± 2.05) decreased by 20% and 50% comparing with contrast group (99.27 ± 1.10, 99.30 ± 0.60) . After treated by Cr (VI) for 24 h, the expression of mRNA and protein level among DNMT1, DNMT3a, DNMT3b and MBD2 decreased significantly in healthy 16HBE cells; and the expression of DNMT1 and DNMT3a decreased in PARG deficiency cells. The relevant expression levels of mRNA of DNMT1 were separately (0.99 ± 0.09), (0.79 ± 0.10), (0.59 ± 0.13) and (0.39 ± 0.02) (F = 247.17, P < 0.01), the expression levels of protein were separately (1.00 ± 0.03), (0.69 ± 0.15), (0.65 ± 0.10) and (0.55 ± 0.13) (F = 214.12, P < 0.01), the expression levels of DNMT3a mRNA were separately (1.00 ± 0.04) , (0.93 ± 0.11) , (0.79 ± 0.07) , (0.59 ± 0.05) (F = 498.16, P < 0.01) , and the expression levels of protein were separately (1.00 ± 0.14) , (0.97 ± 0.11) , (0.79 ± 0.17) , (0.57 ± 0.15) (F = 390.11, P < 0.01) when the dose of Cr (VI) at 0, 0.3, 1.2 and 5.0 µmol/L. However, there were no significant changes of expression found in DNMT3b and MBD2.

CONCLUSIONPoly-ADP-ribosylation could regulate the activity of DNMT3b and MBD2, protect cells against the DNA methylation alteration induced by Cr(VI) and maintain the global genomic DNA methylation level.

Cell Line ; Chromium ; toxicity ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; drug effects ; DNA-Binding Proteins ; metabolism ; Epithelial Cells ; metabolism ; Genome ; Humans ; Poly Adenosine Diphosphate Ribose ; metabolism ; RNA, Messenger ; genetics