Proteomic profiling change during the early development of silicosis disease.

BACKGROUND: Silicosis is one of several severe occupational diseases for which effective diagnostic tools during early development are currently unavailable. In this study we focused on proteomic profiling during the early stages of silicosis to investigate the pathophysiology and identify the proteins involved. METHODS: Two-dimensional (2D) gel electrophoresis and MALDI-TOF-MS were used to assess the proteomic differences between healthy individuals (HI), dust-exposed workers without silicosis (DEW) and silicosis patients (SP). Proteins abundances that differed by a factor of two-fold or greater were subjected to more detailed analysis, and enzyme linked to immunosorbent assay (ELISA) was employed to correlate with protein expression data. RESULTS: Compared with HI, 42 proteins were more abundant and 8 were less abundant in DEW, and these were also differentially accumulated in SP. Closer inspection revealed that serine protease granzyme A, alpha-1-B-glycoprotein (A1BG) and the T4 surface glycoprotein precursor (TSGP) were among the up-regulated proteins in DEW and SP. Significant changes in serine proteases, glycoproteins and proto-oncogenes may be associated with the response to cytotoxicity and infectious pathogens by activation of T cells, positive regulation of extracellular matrix structural constituents and immune response, and fibroblast proliferation. Up-regulation of cytokines included TNFs, interferon beta precursor, interleukin 6, atypical chemokine receptor 2, TNFR13BV, and mutant IL-17F may be involved in the increased and persistent immune response and fibrosis that occurred during silicosis development. CONCLUSIONS: Granzymes, glycoproteins, cytokines and immune factors were dramatically involved in the immune response, metabolism, signal regulation and fibrosis during the early development of silicosis. Proteomic profiling has expanded our understanding of the pathogenesis of silicosis, and identified a number of targets that may be potential biomarkers for early diagnosis of this debilitating disease.

Proteomic profiling differences in serum from silicosis and chronic bronchitis patients: a comparative analysis.

BACKGROUND: Silicosis is a severe occupational disease characterized by pulmonary fibrosis, whereas chronic bronchitis (CB) is an acute inflammation of the airways. Differences in the mechanisms of pathogenesis of these diseases are not well understood, therefore we performed proteomic profiling of silicosis and CB patients and, compared the results. METHODS: Two-dimensional gel electrophoresis and MALDI-TOF-MS (matrix assisted laser desorption ionization time of flight mass spectrometry) were used to identify differentially accumulated proteins in stage I of silicosis (SI), stage II of silicosis (SII) and CB. Enzyme linked immunosorbent assay (ELISA) was employed to validate protein expression data. RESULTS: A total of 28 and 10 proteins were up- and down-regulated in SI, and 21 and 9 proteins were up- and down-regulated SII, compared with CB. Transforming growth factor beta-1 precursor and interferon beta precursor were up-regulated in CB, while interleukin 6, tumor necrosis factor (TNF) and a variant TNF receptor 13B were down-regulated in CB. Additionally, glycoprotein- and apolipoprotein-associated proteins including apolipoprotein A-IV and α-1-B-glycoprotein were up-regulated in CB, indicating an involvement in the pathogenesis of CB but not silicosis. By contrast, HLA-DRB1, medullasin and the proto-oncogene c-Fos were up-regulated in CB. CONCLUSIONS: The immune, metabolism and apolipoprotein-related proteins were identified as playing specific and different roles in silicosis and CB. These proteomic profiling differences would facilitate further studies on the mechanisms underlying silicosis and CB, and may also prove useful to disease diagnosis and treatments.

[Effects of 1-bromopropane on liver and kidney functions of exposed workers].

OBJECTIVE: To study the effects of 1-bromopropane (1-BP) on liver and kidney functions of exposed workers. METHODS: Occupational health situation in three 1-BP plants was investigated. Fifty-four workers from the 1-BP manufacturing line were chose to be contact group, while 42 workers from non-1-BP manufacturing line as control group. All workers underwent questionnaire survey, liver function test as well as kidney function test. RESULT: Working years has no impact on liver and kidney functions of workers from contact group. Compared with the control, liver and kidney functions test of the two groups showed no statistical difference either. CONCLUSION: The present investigation doesn't prove any impact of occupational 1-BP exposure on worker's liver and kidney functions.

[Determination of total Bromine in urine by inductively coupled plasma mass spectrometry (ICP-MS)].

OBJECTIVE: To establish a method to determine total bromine in urine. METHOD: Diluted urine samples were directly introduced into ICP-MS then quantized by standard curve. RESULT: Total bromine in urine was linear within 1.0~50 mg/L with r > 0.999, When spiked at a concentration of 0.020 mg/L, 0.050 mg/L, 0.150 mg/L, the recovery was 95%~98%, intra-assay precision was 1.4% 3.2%, inter-assay precision was 3.4% to 5.0%. Urine could store in -20 °C refrigerator 3 months without any bromine loss. CONCLUSION: Using ICP-MS to determine the urinary total bromine, the method is fast, accurate, wide linear range of features, could meet with the requirement of Part 5 of occupational health standards guide: Method determination of chemical substances in biological materials (GBZ/T 210.5-2008), a strong competitive advantage in a wide range of survey, suitable for promotion.

[Comparative analysis of serum proteomic profiles between patients with silicosis and chronic bronchitis].

OBJECTIVE: To analyze the differences in serum proteomic profiles between patients with silicosis and chronic bronchitis and to investigate the pathogenesis, clinical diagnosis, and treatment of these two disease. METHODS: Serum samples from patients with stage I silicosis and chronic bronchitis were collected. Two-dimensional gel electrophoresis was performed and protein plots with expression differences higher than 2-fold were identified and further analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Compared with the silicosis group, the chronic bronchitis group had 11 up-regulated proteins and 23 down-regulated proteins. The chronic bronchitis group had high expression of proteins such as interferon beta precursor, apolipoprotein precursor, and transforming growth factor beta1 precursor. The silicosis group had high expression of proteins such as interleukin-6, granzyme A, cathepsin G, and glycoprotein precursor. CONCLUSION: The differentially expressed proteins are mainly involved in the activity of serine enzymes, cytotoxicity, inflammation response, and apolipoprotein transfer and play different roles in silicosis and chronic bronchitis.

[Changes in serum protease and cytokine in patients with silicosis, tuberculosis, and lung cancer].

OBJECTIVE: To investigate the changes in serum protease and cytokine in patients with silicosis, tuberculosis, and lung cancer. METHODS: Serum samples of patients with silicosis, tuberculosis, and lung cancer were collected. The variation trends of the expression of granzyme A, cathepsin G, apolipoprotein A, and interferon-ß (IFN-ß) were analyzed using enzyme-linked immunosorbent assay. RESULTS: The concentration of apolipoprotein A of the silicosis group was 200 µg/ml, significantly higher than those of the tuberculosis and lung cancer groups (P < 0.05), and the lung cancer group had a significantly higher concentration of apolipoprotein A compared with the tuberculosis group (P < 0.05). The silicosis group had significantly higher expression of cathepsin G compared with the tuberculosis and lung cancer groups (P < 0.05), and the tuberculosis group and lung cancer group showed no significant difference in the concentration of cathepsin G (P > 0.05). The tuberculosis group had a significantly higher concentration of granzyme A than the silicosis and lung cancer groups (P < 0.05), and the silicosis group and lung cancer group had similar protein concentration trends (P > 0.05). The tuberculosis group and lung cancer group had significantly higher concentration of IFN-ß compared with the silicosis group (P < 0.05), and the tuberculosis group and lung cancer group showed no significant difference in IFN-ß concentration (P > 0.05). CONCLUSION: This study may offer diagnostic markers for the clinical diagnosis of silicosis, tuberculosis, and lung cancer, and could provide a basis for the research, as well as potential molecular targets for the diagnosis and treatment of these diseases.

[Clinical efficacy of tetrandrine combined with acetylcysteine effervescent tablets in treatment of silicosis].

OBJECTIVE: To observe the clinical efficacy of tetrandrine combined with acetylcysteine effervescent tablets in the treatment of silicosis. METHODS: A total of 96 patients with silicosis were randomly divided into treatment group (49 cases) and control group (47 cases). Both groups were given routine therapy including anti-inflammatory, antitussive, and antiasthmatic drugs, and the patients in treatment group were given tetrandrine combined with acetylcysteine effervescent tablets at the same time. Tetrandrine (100 mg) was orally administrated twice a day, and there was a one-day interval between every 6 days' continuous administration; totally, there were four courses of treatment, with 3 months for each course, and there was a one-month break between each course. Acetylcysteine effervescent tablets (600 mg) were taken twice a day; each course of treatment was 12 days, and there were four courses; for the first two months, there was one course per month, and then one course every other two months for the rest of time. Clinical symptoms, pulmonary ventilation function, serum superoxide dismutase (SOD) and changes in X-ray findings were observed. RESULTS: After treatment, the treatment group had significantly increased rates of improvements in cough, expectoration, chest congestion and pain, and dyspnea compared with the control group (P < 0.05). Compared with the control group (serum SOD level: 70.466±20.261 U/ml) and the treatment group before therapy (serum SOD level: 68.182±21.414 U/ml), the treatment group after therapy had significantly increased serum SOD level (77.389±21.315 U/ml?, forced vital capacity, and forced expiratory volume in one second (P < 0.05). Eight patients in treatment group showed improvement in the chest X-ray findings of silicosis. CONCLUSION: The combination of tetrandrine and acetylcysteine effervescent tablets show some effect in the treatment of silicosis. It can be an effective option for treating silicosis as there are no other specific remedies.

[Changes in activities of SOD and GSH-Px in induced sputum and their significance among silicosis patients].

OBJECTIVE: To observe the changes in activities of superoxide dismutase (SOD) and glutathione peroxide (GSH-Px) in the induced sputum of silicosis patients, and to investigate the roles of SOD and GSH-Px in the development and progression of silicosis and the significance of measuring activities of SOD and GSH-Px in induced sputum among silicosis patients. METHODS: Fifty hotel attendants were chosen as control group, 50 workers with more than one year of silica dust exposure as dust exposure group, 32 silica dust-exposed workers as observation subject group, and 52 silicosis patients as silicosis group. The activities of SOD and GSH-Px in their induced sputum were measured by enzyme-linked immunosorbent assay. RESULTS: Compared with the control group, the observation subject group and silicosis group had significantly decreased SOD activity (68.16 ± 30.17 and 66.38 ± 47.32 U/ml vs 75.81 ± 11.92 U/ml, P < 0.05); compared with the dust exposure group, the silicosis group had significantly decreased SOD activity (66.38 ± 47.32 U/ml vs 70.12 ± 14.31 U/ml, P < 0.05). Compared with the control group and dust exposure group, the observation subject group and silicosis group had significantly increased GSH-Px activity (268.21 ± 15.45 and 279.34 ± 29.26 U/ml vs 224.22 ± 12.64 and 236.41 ± 14.54 U/ml, P < 0.05 or P < 0.01). CONCLUSION: The SOD activity in dust exposure group and silicosis group decreased, but there were no significant differences between patients with different stages of silicosis. The GSH-Px activity in dust exposure group and silicosis group was significantly higher than that in control group, and there were significant differences between patients with different stages of silicosis. These suggest that the imbalance of oxidative/antioxidant systems is associated with the development and progression of silicosis.

[Determination of 2, 4-toluenediamine in urine by gas chromatography].

OBJECTIVE: To establish a method for determining the content of 2,4-toluenediamine, a urinary metabolite of toluene diisocyanate, by gas chromatography. METHODS: Urine samples were collected, and acidification, extraction, derivatization, separation with a capillary column, and detection with an electron capture detector were performed. The target compound was qualified by retention time and quantified by peak area. RESULTS: The concentration of 2, 4-toluenediamine showed a linear relationship with peak area within 0.0∼40 ng/ml, with a correlation coefficient 0.9995; the limit of detection was 0.44 ng/ml, the lower limit of quantification was 1.47 ng/ml, the relative standard deviation was 1.85%∼4.05%; the recovery rate was 97.98%∼99.28%. CONCLUSION: The method has the advantages of high sensitivity and high accuracy and can be used for determination of 2, 4-toluenediamine in urine.

[Therapeutic efficacy of tetrandrine tablets combined with matrine injection in treatment of silicosis].

OBJECTIVE: To observe the therapeutic efficacy of tetrandrine tablets combined with matrine injection in the treatment of silicosis. METHODS: Sixty-three patients with silicosis were randomly divided into treatment group (n = 33) and control group (n = 30). Both groups received anti-inflammatory, cough-relieving, and anti-asthmatic treatment. Meanwhile, the treatment group was given tetrandrine tablets (100 mg bid) and matrine injection (150 mg qd). There were 4 courses of tetrandrine treatment (each course = 3 months), with one-month intervals among them. Matrine injection was used for 15 consecutive days in each month. There were 2 courses of matrine treatment (each course = 3 months), with a one-month interval in between. The clinical symptoms, pulmonary function, serum superoxide dismutase (SOD) activity, and chest X-ray images were observed before and after treatment. RESULTS: After treatment, chest distress, chest pain, shortness of breath, and other respiratory symptoms were relieved significantly (P < 0.05). The treatment group showed significantly higher SOD activity than before treatment and the control group (P < 0.05) and significantly higher forced vital capacity and forced expiratory volume in one second than before treatment and the control group (P < 0.05). After treatment, 5 patients (4 stage II cases and 1 stage III case, all in rapidly progressive forms) in the treatment group showed smaller, lighter, and clearer shadows with decreased overall intensity on chest X-ray; 12 patients showed significantly fewer and clearer lung markings on chest X-ray. CONCLUSION: Tetrandrine tablets combined with matrine injection have some therapeutic effect on silicosis.