The clinical value of induced sputum DKK3 gene methylation in the evaluation of disease condition and prognosis for non-small cell lung cancer / 中国医师进修杂志

Objective:To study the clinical value of induced sputum DKK3 gene methylation in the evaluation of disease condition and prognosis for non-small cell lung cancer (NSCLC).Methods:Eighty NSCLC patients (observation group) and 50 benign lung disease patients (control group) who were treated in Linshu County People′s Hospital of Shandong Province from January 2015 to December 2017 were enrolled. Methylation specific polymerase chain reaction (PCR) was used to detect the induced sputum DKK3 gene methylation. The DKK3 gene methylation rates in different clinicopathological factors were compared.Results:The DKK3 gene methylation rate in observation group was significantly higher than that in control group: 81.3%(65/80) vs. 2.0%(1/50), the difference was statistically significant ( P<0.05). The DKK3 gene was methylated in lung cancer cells, and was unmethylated in normal lung epithelial cells BEAS-2B. The DKK3 gene methylation rates had correlation with pleural effusion, degree of differentiation, tumor diameter, lymph node metastasis, distant metastasis and TNM staging ( P<0.05). The R0 radicalresection, 3-year survival rate and total survival time in patients with DKK3 gene methylated were significantly lower than those in patient with DKK3 gene unmethylated: 53.8%(35/65) vs. 15/15, 28.1% vs. 37.9%, (1.8 ± 0.3) years vs. (2.1 ± 0.6) years, the differences were statistically significant ( P<0.05). Conclusions:The induced sputum DKK3 gene methylation rate in NSCLC patients is significantly higher and is related with prognosis. The induced sputum DKK3 gene methylation may provide basis for evaluating of disease condition and prognosis for NSCLC patients.

Gene variance in microsomal epoxide hydrolase and the susceptibility of coal workers' pneumoconiosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore whether the tagging single nucleotide polymorphisms (SNPs) within EPHX1 gene were involved in the genetic susceptibility to coal worker's pneumoconiosis (CWP) by case-control study.</p><p><b>METHODS</b>This study consisted of 697 CWP patients and 694 controls. All the subjects were Han Chinese, underground coal miners and recruited from coal mines of Xuzhou Mining Business Group Co Ltd.. The venous blood samples were obtained from all subjects and extracted genome DNA from the isolated leucocytes. Three SNPs were selected from the HapMap and the genotyping was done by the TaqMan method with the ABI 7900HT Real Time PCR system.</p><p><b>RESULTS</b>The Single SNP analyses showed that the genotype frequencies of EPHX1 (rs2234922) was significantly associated with decreased risk of CWP under co-dominant model (OR = 0.22, 95% CI = 0.06~0.79, P = 0.020), recessive model (OR = 0.23, 95% CI = 0.06~0.82, P = 0.023), and addictive model (OR = 0.75, 95% CI = 0.58~0.96, P = 0.022). The further stratification analysis showed that the risk of CWP will significantly decreased in non-smoking groups (OR = 0.10, 95% CI = 0.01~0.83, P = 0.033).</p><p><b>CONCLUSIONS</b>Our results suggest that individuals with the EPHX1 (rs223492) GG genotype was associated with a dereased risk of CWP, and it has a protective effect on the developing CWP.</p>

Regulatory effect of miR-149 on interleukin-6 expression in silica-induced pulmonary fibrosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the regulatory effect of miR-149 on interleukin-6 (IL-6) expression in silica-induced pulmonary fibrosis.</p><p><b>METHODS</b>A mouse model of pulmonary fibrosis was established using silica dust; the level of miR-149 in the lung tissues of mice with silica-induced pulmonary fibrosis was measured by quantitative real-time polymerase chain reaction (qRT-PCR), while the protein expression of IL-6 was measured by immunohistochemistry and Western blot. Type II alveolar epithelial cells (A549) and bronchial epithelial cells (HBE) were exposed to silica dust to establish a model; the level of miR-149 was measured by qRT-PCR, while the protein expression of IL-6 was measured by Western blot. A549 cells were transfected with miR-149 mimics and inhibitor in vitro, and the cellular expression of IL-6 was measured by Western blot. Serum samples from patients with coal workers' pneumoconiosis were examined by double-antibody sandwich ELISA to measure the protein expression of IL-6.</p><p><b>RESULTS</b>At three time points after silica treatment, the miR-149 expression in lung tissues was significantly down-regulated while an evident increase in IL-6 expression was observed in lung tissues (P < 0.01). Silica-stimulated epithelial cell (A549 and HBE) had up-regulated IL-6 expression and down-regulated miR-149 expression (P < 0.01). Increased levels of miR-149 attenuated IL-6 expression, whereas adverse results were found when miR-149 was inhibited. Compared with that in control group, serum level of IL-6 was significantly increased in patients with stage II and III coal workers' pneumoconiosis (P < 0.01).</p><p><b>CONCLUSION</b>Down-regulation of miR-149 and up-regulation of IL-6 might be involved in the progression of silica-induced pulmonary fibrosis; miR-149 could negatively regulate IL-6 expression.</p>

Regulation of melanin expression in both cell and gene by retinoic acid / 中华医学美学美容杂志

Objective To investigate the effects and mechanism of retinoie acid on B16 marina mel-anoma cells and human melanocytes in vitro. Methods B16F10 murine melanoma cells and human mela-noeytes were cultured in culture medium which contains different concentration of components, including retinoic acid. Using reverse transcription-polymerase chain reaction (RT-PCR) mRNA expression of the tyrosinase was detected. Tyrosinase activity, melanin content and cell proliferation rate were also deter-mined. Results Retinoieacid exhibited an inhibitory effect on the expression of tyrosinase mRNA. As the concentration of retinoic acid was 100 μmol/L, treating for 72 h, the expression of tyrosinase mRNA de-creased 30.13 %, retinoic acid exhibited an inhibitory effect on tyrosinase activity and melanin production at high concentration (>500 μmol/L), and it could promote the cell proliferation. Retinoic acid and hy-droqninone could be cooperative at high concentration (1 000 μmol/L), and enhanced the down regulation of tyrosinase activity and melanin content. Retinoic acid could also mitigate the inhibitory effect of hydro-quinone on cell proliferation, so as to protect the cells from injury. Hydroquinone had no effect on tyrosi-nase gene expression at mRNA level. Conclusion Retinoic acid inhibits the synthesis of melanin by the genetic regulation at mRNA level.