Genetic polymorphisms and surface expression of CTLA-4 and PD-1 on T cells of silica-exposed workers

Exposure to silica dust has been examined as a possible risk factor for autoimmune diseases, including scleroderma, rheumatoid arthritis and systemic lupus erythematosus. Since CTLA-4 [CD152] and PD-1 [CD279] are important for the maintenance of peripheral tolerance by regulating T cell responsiveness, we evaluated the expression of these molecules on the surface of CD4 and CD8 T cells, as well as single nucleotide polymorphisms (SNP) in CTLA-4 and PDCD1 genes, of 70 silica-exposed workers and 30 non-exposed, age-, ethnically- and sex-matched controls. Expression of CTLA-4 was significantly (P<0.05) reduced in CD4 T cells of exposed individuals [median=0.1% and interquartile range, IQR 0.0-0.1% (exposed), median=0.20%, IQR 0.0-0.4% (control)]. Also the expression of PD-1 was significantly (P<0.0001) reduced in both CD4 [median=0.9%, IQR 0.4-2.3% (exposed), median=5.7%, IQR 1.4-13.3% (control)] and CD8 T cells [median=0.9%, IQR 0.3-1.9% (exposed), median=5.0%, IQR 3.4-8.9% (control)]. The study of polymorphisms demonstrated a lower frequency of the A allele in the analysis of the PD1.3 SNP in the exposed group, which might be associated with the lower expression of PD-1 on the surface of CD4 T cells. Our findings provide evidence for the association of silica exposure and the maintenance of self-tolerance, i.e., the susceptibility to autoimmune disorders.     

SiO_2粉尘刺激对NR8383细胞分泌炎症因子的影响

目的:比较不同剂量Si O2粉尘刺激NR8383细胞后TNF-α,IL-1β,TGF-β基因表达水平变化。方法:细胞接受不同剂量粉尘刺激后,采用MTT法检测各组细胞活力的改变,筛选出细胞存活率比较高的剂量。细胞随机分为四组,即对照组,10μg/cm2组、20μg/cm2组和40μg/cm2组,接受粉尘刺激12 h后,采用荧光定量PCR法测定各组TNF-α,IL-1β,TGF-β基因表达水平。结果:随着染尘剂量的提高,细胞存活率逐渐降低,80μg/cm2组细胞存活率最低。实时荧光定量PCR显示10μg/cm2组、20μg/cm2组和40μg/cm2组TNF-α,IL-1β,TGF-β基因表达水平均高于对照组,差异具有统计学意义(P<0.05)。随着染尘剂量的递增,TNF-α,IL-1β,TGF-β的mRNA相对表达水平逐渐增高。结论:粉尘刺激可以引起大鼠肺泡巨噬细胞分泌炎症因子增加,呈现一定的剂量效应关系。     

A study of the prevalence of respiratory morbidity among agate workers

Summary A random sample of 342 workers engaged in chipping and grinding of agate stones were surveyed in a cross-sectional study to assess the prevalence of respiratory morbidity in the agate industry. The findings were compared with those obtained in controls. The study showed a significantly higher prevalence of lung diseases among agate workers than among controls (63.4% vs 35.5%, P < 0.001). The respiratory morbidity in agate workers and controls was maximal in the group aged 31+ years (83.3% and 42.2%, respectively). The prevalence of pneumoconiosis in agate workers (18.4%) was highly significant as compared with controls, in whom not a single case was found (P < 0.001). However, there was no significant difference between the prevalence of pneumoconiosis in men and women of the exposed group (17.9% vs 19.6%). Among the cases of pulmonary diseases in agate workers, pneumoconiosis formed the largest group (18.4%), whereas among controls it was tuberculosis (12.1%). The prevalence of pulmonary tuberculosis was very high in both agate workers and controls (15.5% and 12.1%, respectively), probably because of poor socio-economic and unhygienic living conditions. The prevalence of chronic bronchitis was found to be higher among the control population as compared with the exposed group (6.7% vs 2.6%). It therefore appeared that agate dust had no role in precipitating chronic bronchitis. However, bronchial asthma appeared to have been aggravated due to agate dust, as the risk among agate workers was 7-fold that found among the controls. The prevalence of pneumoconiosis showed a dose-response relationship in both male and female agate workers. Pulmonary tuberculosis and bronchial asthma did not show any significant trend in relation to the duration of exposure. Other lung diseases were also independent of exposure. As far as occupations are concerned, grinders were found to suffer more from pneumoconiosis (21.9%) than did chippers (7.3%, P < 0.05); this was because the respirable dust concentration at the workplace of grinders was 6-fold that at the workplace of chippers. Acute bronchitis was also more prevalent in grinders (P < 0.05). However, tuberculosis and asthma did not vary significantly between the two occupational groups.     

二氧化硅对人红细胞膜ATPase和乙酰胆碱脂酶的影响

本研究探讨了二氧化硅粉尘对人红细胞膜乙酰胆碱脂酶(AChE)、Na~+、K~+-ATPase 和 Ca~(2-)-ATPase 的相互作用。结果显示二氧化硅具有吸附 AChE 的能力,对 Ca~(2+)-ATPase 活性有一定抑制,而 Na~+、K~+-ATPase 活性有所上升。提示二氧化硅在损伤细胞膜的过程中对膜酶也会产生影响。     

In vitro toxicity of silica nanoparticles in human lung cancer cells

The cytotoxicity of 15-nm and 46-nm silica nanoparticles was investigated by using crystalline silica (Min-U-Sil 5) as a positive control in cultured human bronchoalveolar carcinoma-derived cells. Exposure to 15-nm or 46-nm SiO(2) nanoparticles for 48 h at dosage levels between 10 and 100 microg/ml decreased cell viability in a dose-dependent manner. Both SiO(2) nanoparticles were more cytotoxic than Min-U-Sil 5; however, the cytotoxicities of 15-nm and 46-nm silica nanoparticles were not significantly different. The 15-nm SiO(2) nanoparticles were used to determine time-dependent cytotoxicity and oxidative stress responses. Cell viability decreased significantly as a function of both nanoparticle dosage (10-100 microg/ml) and exposure time (24 h, 48 h, and 72 h). Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species (ROS), glutathione, malondialdehyde, and lactate dehydrogenase, were quantitatively assessed. Exposure to SiO(2) nanoparticles increased ROS levels and reduced glutathione levels. The increased production of malondialdehyde and lactate dehydrogenase release from the cells indicated lipid peroxidation and membrane damage. In summary, exposure to SiO(2) nanoparticles results in a dose-dependent cytotoxicity in cultural human bronchoalveolar carcinoma-derived cells that is closely correlated to increased oxidative stress.     

Development of a chronic inhalation reference level for respirable crystalline silica

Chronic inhalation exposure of workers to crystalline silica can result in silicosis. The general public can also be exposed to lower levels of crystalline silica from quarries, sand blasting, and entrained fines particles from surface soil. We have derived an inhalation chronic reference exposure level for silica, a level below which no adverse effects due to prolonged exposure would be expected in the general public. Incidence of silicosis and silica exposure data from a cohort of 2235 white South African gold miners yielded a reference level of 3 microg/m3) for respirable silica (particle size as defined occupationally) using a benchmark concentration approach. Data from cohorts of American gold miners, Chinese tin miners, diatomaceous earth workers, and black South African gold miners yielded similar results with a range of 3-10 microg/m3. Strengths of the chronic reference exposure level include the availability of several large long-term studies of inhalation in workers at varying exposure concentrations, adequate histopathological and radiologic analysis, adequate follow-up of exposed workers, a dose-response effect in several studies, observation of a No Observed Adverse Effect Level in the key study, and the power of the key study to detect a small effect. Uncertainties include the general underestimation of silicosis by radiography alone and the uncertainties in exposure estimation.     

Influence of synthesis and processing conditions on the release behavior and stability of sol-gel derived silica xerogels embedded with bioactive compounds

The influence of processing parameters and synthetic strategies in the properties of sol-gel derived silica matrices intended for the release of bioactive compounds was investigated. The time-evolution of the matrix properties during its aging at room temperature in the dry and wet forms was investigated by measuring some of its physical and drug retaining properties. The results indicate that long term gel aging in the wet form is fundamental for the obtainment of dry matrices that are stable upon storage, a fundamental requirement for any practical application. In the case of hybrid matrices obtained by replacing part of the tetraethoxysilane precursor with mono-methyl trimethoxysilane, the order of addition of the reaction component is also important in determining the properties of the final dry gel, probably by influencing the polymer structural properties. This parameter acts synergistically with the matrix composition in determining the release properties of xerogels embedded with bioactive compounds.     

SiO2粒径分布对牙科复合树脂性能影响

目的研究SiO2粒径分布对牙科复合树脂无机填料的填充量和物理机械性能的影响。方法采用经硅烷表面修饰的气相法纳米SiO2(NS)和微米SiO2(MS),通过调整两者相对质量比,与双酚A双甲氧基缩水甘油酯牙科树脂单体共混,制备了一系列牙科复合树脂M50N20,M55N15,M58N12,M60N10,M63N07,研究了不同SiO2粒径分布对无机填料的填充量,以及复合树脂维氏硬度、三点弯曲强度和断面形貌的影响。结果复合树脂中填料的填充量取决于无机填料的分布及比表面大小,当MS/NS=58/12时,复合树脂的无机填料填充量达到84wt%。M58N12表现出最高的三点弯曲强度(153.2MPa)以及维氏硬度(102.3MPa)。复合树脂断面的SEM形貌分析表明,纳米SiO2与复合树脂基体有较好的粘合作用。结论复合树脂中SiO2粒径分布对物理机械性能以及无机填料的填充量有较显著的影响。     

Roles of Rho/Rock Signaling Pathway in Silica-induced Epithelial-mesenchymal Transition in Human Bronchial Epithelial Cells

ObjectiveTo investigate the roles of Rho/Rock signaling pathway in silica-induced Epithelial-mesenchymal transition (EMT) in human bronchial epithelial cells (BEC) in vitro.MethodsHuman BEC were incubated with silica with various concentrations for indicated times. Cell viability was assayed by MTT test. Morphologic Changes were observed by microscope. Mesenchymal marker α-smooth muscle actin (α-SMA), vimentin (Vim), and epithelial marker E-cadherin (E-cad) were analyzed by Western Blot. The pull-down assay was used to measure Rho activity. In the prevention experiments, the specific inhibitor for Rho effector ROCK (Y27632) was used to inhibit the activity of Rho.ResultsHuman BEC stimulated with silica were converted from a “cobblestone” epithelial structure into an elongated fibroblast-like shape structure. Incubation of human BEC with silica induced de novo expression of α-SMA and Vim, and loss of E-cad. Also, silica treatment resulted in Rho activation in human BEC. Y27632 up-regulated the E-cad expression but attenuated α-SMA and Vim expression in silica-stimulated cells.ConclusionThe activation of Rho/ROCK signaling pathways is most likely involved in Silica-induced EMT in human bronchial epithelial cells.     

Bone morphogenetic protein-7 inhibits silica-induced pulmonary fibrosis in rats

Bone morphogenetic protein-7 (BMP-7) has been shown to inhibit liver and renal fibrosis in in vivo and vitro studies. There is no study to investigate BMP-7's role in the development of pulmonary fibrosis induced by silica. In the current study, we used the rat model to explore the potential antifibrotic role of BMP-7 and its underlying mechanism in silica-induced pulmonary fibrosis. Sixty Wistar rats were randomly assigned into three groups. Control group received saline, silica group received silica and BMP-7 treated group received silica and BMP-7. BMP-7 was administered to silica-treated rats intraperitoneally at a dose of 300 μg/kg/injection from day 8 to day 30 every other day. After the animals were sacrificed on day 15 and 30, hydroxyproline levels, the protein expressions of BMP/Smad and TGF-β/Smad signaling, and histopathology in lung tissues were analyzed. The hydroxyproline contents in BMP-7 treated groups were significantly lower than the silica groups (P < 0.05). Histopathological results showed BMP-7 could reduce the progression of silica induced fibrosis. Furthermore, the expression of p-Smad1/5/8, a marker of BMP/Smad signaling, was significantly up-regulated in BMP-7 treated groups (P < 0.05) compared with the silica groups. On the contrary, the expression of p-Smad2/3, a marker for TGF-β/Smad signaling, reduced significantly in BMP-7-treated groups compared with silica groups (P < 0.05). In conclusion, the pulmonary fibrosis induced by silica in rats was significantly reduced with the therapeutic treatment of BMP-7. The antifibrotic effect of BMP-7 could be related to the activation of BMP/Smad signaling and inhibition of TGF-β/Smad pathways.