Type I IFNs repolarized a CD169+ macrophage population with anti-tumor potentials in hepatocellular carcinoma.

Macrophages constitute a major component in human hepatocellular carcinoma (HCC) and perform various functions to facilitate disease progression. Reprogramming or reconstituting the tumor surveillance phenotypes of macrophages represents an attractive immunotherapeutic strategy in cancer treatments. The current study identified CD169 as a potential target for macrophage repolarization since it signified a population of macrophages positively correlated with an activated immune signature and better prognosis of patients with HCC. In vitro experiments revealed that a low dose of type I interferon (IFN) could effectively reprogram human monocyte-derived macrophages to upregulate CD169 expression, and such induced CD169+ macrophages exhibited significantly enhanced phagocytotic and CD8+ T cell-activating capacities compared to controls. A low dose of IFNα also inhibited hepatoma growth in mice in vivo, presumably through polarizing the CD169+ macrophage population and enhancing CD8+ T cell activities. Notably, IFNα also induced substantial PD-L1 expression on macrophages in vivo, and thus blockade of PD-L1 could further increase the anti-tumor efficacy of IFNα in the treatment of HCC. We propose a low dose of IFNα in combination with a PD-L1 blocking agent as a potential anti-tumor therapeutic strategy via its effects on macrophage polarization.

High S100A9+ cell density predicts a poor prognosis in hepatocellular carcinoma patients after curative resection.

S100A9 is differentially expressed in various cell types and is associated with the development, progression and metastasis of various cancers. However, the expression, distribution, and clinical significance of S100A9 in hepatocellular carcinoma (HCC) remain unclear. In the present study, The Cancer Genome Atlas (TCGA) database was used to examine S100A9 gene expression in HCC; we found that S100A9 expression was associated with HCC prognosis. In addition, S100A9 protein expression was assessed by immunohistochemistry analysis of tissues from 382 HCC patients. We found that the infiltration of S100A9+ cells in both tumor and nontumor tissues could predict poor overall survival (P = 0.0329, tumor; P = 0.0003, nontumor) and a high recurrence risk (P = 0.0387, tumor; P = 0.0015, nontumor) in our tissue microarray analysis. Furthermore, immunofluorescence double staining revealed that the primary S100A9-expressing cells in adjacent nontumoral tissue were CD15+ neutrophils, and both CD68+ macrophages and CD15+ neutrophils expressed S100A9 in HCC tumor tissues. Taken together, the results suggest that high S100A9+ cell density predicts a poor prognosis in HCC patients, and S100A9 expression could potentially serve as an independent prognostic marker for HCC.

Targeting adenosinergic pathway enhances the anti-tumor efficacy of sorafenib in hepatocellular carcinoma.

BACKGROUND: Sorafenib is the most widely used first-line treatment for patients with advanced hepatocellular carcinoma (HCC), but such treatment provides only limited survival benefits that might be related to the immune status of distinct tumor microenvironments. A fundamental understanding of the distribution and phenotypes of T lymphocytes in tumors will undoubtedly lead to the development of novel immunotherapeutic strategies that could possibly enhance the efficacy of sorafenib treatments. METHODS: Flow cytometry, immunohistochemistry and immunofluorescence analyses were performed to detect the infiltration and distribution of various leukocyte populations, and the expression of different immune checkpoint molecules in fresh HCC tumor tissues. Correlations among indicating genes were calculated in 365 patients with HCC from The Cancer Genome Atlas (TCGA) data set, and the cumulative overall survival time was calculated using the Kaplan-Meier method. Moreover, role of adenosinergic pathway on sorafenib anti-tumor efficacy was investigated using both subcutaneous and orthotopic transplantation tumor model in immune competent C57BL/6 mice. RESULTS: We revealed that levels of CD3+ and CD8+ T cells were significantly downregulated in HCC tumor tissue, so were the infiltration of CD169+ cells (a Mφ subpopulation with T cell activation capacities) and their contact with CD8+ cells in tumor milieus. Moreover, levels of PD-1 and CD39 expression were significantly upregulated in human HCC-infiltrating CD4+ and CD8+ T cells, and CD39+CD8+ T cells exhibited a CD69+PD-1+perforinlowIFNγlow "exhausted" phenotype. Levels of both CD39+ T cells infiltration and adenosine receptor ADORA2B expression in tumor tissues were negatively correlated with overall survival of patients with HCC. Accordingly, mice treated with sorafenib in combination with adenosine A2B receptor blockage reagents exhibited significantly reduced tumor progression compared with control groups. CONCLUSIONS: These results suggest that adenosinergic pathway might represent an applicable target for sorafenib-combined-therapies in human HCC.

An analysis on plasma microRNA profiling and function among population exposed to asbestos / 预防医学

Objective To investigate the effects of asbestos exposure on plasma miRNA expression.Methods Plasma samples were collected from control group and asbestos -exposed group (time of exposure >10 years)and three samples from each group were selected to detect differentially expressed miRNA using LC Sciences miRNA Microarray -Single.The target genes of differential miRNA were predicted by three kinds of online software,Target Scan,miRanda and PicTar.GO term enrichment and KEGG pathways were analyzed.Results The results of microarray indicated that there were 40 differential miRNA expression between exposed and control groups(P <0.05),and the signal value of 9 differential miRNA exceeded 500.After analyzing signal pathways of target genes of 5 miRNA,of which the signal values were over 500,these target genes were found mainly involved in pathways associated with cancer and metabolism,including potential function targets of FAS,TP53 and FGFR3.Conclusion Asbestos exposure can result in differentially expressed miRNA in the plasma from workers occupationally exposed to asbestos and the target genes of these miRNA may play important roles in the pathways of cancer.However,the mechanism of these miRNA in asbestos -related diseases needs to be further studied in the future.

Effects of p38 mitogen-activated protein kinases on the apoptosis of human bronchial epithelial cells induced by refractory ceramic fibers in vitro / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the role of p38 mitogen-activated protein kinases (MAPKs) in the apoptosis of human bronchial epithelial cells (BEAS-2B) induced by refractory ceramic fibers (RCFs).</p><p><b>METHODS</b>BEAS-2B cells were exposed to 10, 20, 40, 80, and 160 µg/cm(2) RCF1, RCF2, and RCF3 for 24 h, and the cell viability was measured by CCK-8 assay. BEAS-2B cells were exposed to 20, 40, and 100 µg/cm(2) RCF1, RCF2, and RCF3 for 24 h, and the cell apoptosis rate was measured by flow cytometry. BEAS-2B cells were exposed to 40 µg/cm(2) RCF1, RCF2, and RCF3, and the expression levels of phospho-p38 MAPK and caspase-3 were measured by Western blot. In each of the above treatments, the BEAS-2B cells were divided into positive control, p38 inhibitor SB203580 intervention, and normal groups.</p><p><b>RESULTS</b>As the concentration of RCFs rose, the RCF exposure groups showed decreased cell viability and increased cell apoptosis rate. After SB203580 intervention, the intervention groups (all concentrations of asbestos + SB, 20, 40, 80, and 160 µg/cm(2)RCF1+SB, and 40, 80, and 160 µg/cm(2) RCF2 and RCF3+SB) had significantly increased cell viabilities (P < 0.05), and the intervention groups (asbestos + SB and 20, 40, and 100 µg/cm(2) RCF1, RCF2, and RCF3 + SB) had significantly decreased cell apoptosis rates (P < 0.05). Compared with the normal group, the RCF (40 µg/cm(2)) exposure and positive control groups had significantly increased expression of phospho-p38 MAPK (P < 0.05), and the RCF (40 µg/cm(2)) exposure group had significantly increased expression of caspase-3 (P < 0.05). The intervention groups (asbestos + SB and 40 µg/cm(2) RCF1, RCF2, and RCF3 + SB) had significantly decreased expression of caspase-3 after SB203580 intervention.</p><p><b>CONCLUSION</b>p38 MAPKs play an important role in RCF-induced apoptosis of BEAS-2B cells.</p>

Screening of differentially expressed serum proteins in patients with asbestosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To screen differently expressed proteins for serum biomarkers by studying serum proteome of population with asbestosis, population exposed to asbestos without asbestosis and population never exposed to asbestos, to further understand the mechanisms of asbestosis.</p><p><b>METHODS</b>The subjects of present study included 37 patients with asbestosis, 254 workers exposed to asbestos and 439 healthy controls. The 2-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-MS/MS) were used to screen and identify the differentially expressed serum proteins among all subjects. ImageMaster6.0 software was utilized to analyze the differentially expressed proteins.</p><p><b>RESULTS</b>Well-qualified gel images of serum proteome were obtained, 21, 34 and 32 differentially expressed spots were found between asbestosis and normal controls, between asbestosis and negative controls or between negative controls and normal controls, respectively. Differentially displayed proteins were identified as cytokines, α1-AT, L-ficolin, etc.</p><p><b>CONCLUSION</b>Exposure to asbestos for a long period could interfere with the immune system of workers exposed to asbestos, and some proteins may serve as the biomarkers for early diagnosis and intervention of asbestosis.</p>

The protein expression profiles induced by trimethyltin chloride in Vero cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the biomarkers and mechanism of kidney toxicity induced by trimethyltin chloride (TMT-Cl) through analyzing the differences of protein expression profiles between vero cells and vero cells exposed to TMT-Cl.</p><p><b>METHODS</b>The differences of protein expression levels of three paired samples of vero cells and vero cells exposed to TMT-Cl were compared by two-dimensional gel electrophoresis (2-DE) and liquid chromatography-electrospray ionization-linear trap quadrupole (LC-ESI-LTQ). The differences of expression levels of Annexin A1 and α-Tubulin proteins were validated with western blot assay, and the differences of mRNA expression levels of Annexin A1 and α-Tubulin genes were detected with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).</p><p><b>RESULTS</b>Fifteen spots of differential expression in protein profiles between vero cells and vero cells exposed to TMT-Cl were found, and 9 of these spots were identified by LC-ESI-LTQ. The expression levels of 3 proteins (Annexin A1,similar to RAN protein and a hypothetical protein) increased and the expression levels of 6 proteins(growth factor receptor-bound protein 10, tubulin alpha 6, heterogeneous nuclear ribonucleoprotein, similar to elongation factor SIII p15 subunit, S-adenosylhomocysteine hydrolase and a hypothetical protein) reduced. The expression levels of α-Tubulin protein and mRNA significantly decreased in vero cells exposed to TMT-Cl, as compared with vero cells (P < 0.01). The expression of Annexin A1 protein in all exposure groups was significantly up-regulated, the expression of Annexin A1 mRNA in the groups exposed to 25 and 50 µmol/L TMT-Cl was significantly down-regulated, and The expression of Annexin A1 mRNA in the group exposed to 100 µmol/L TMT-Cl was significantly up-regulated (P < 0.01).</p><p><b>CONCLUSIONS</b>The results of present study suggest that 9 proteins with differential expression detected by LC-ESI-LTQ may be related to the kidney toxicity induced by TMT-Cl, which can serve as the biomarkers of early diagnosis and therapeutic effect for the kidney toxicity induced by TMT-Cl.</p>

Pirfenidone diminishes SiO2 induced lung fibrosis in rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>to investigate whether pirfenidone (PFD) presents the antifibrotic effect in silicosis of rats.</p><p><b>METHODS</b>SD rats were randomly divided into five groups: the non-treat group, the normal saline group, the normal saline + PFD group, the SiO2 group, the SiO2 + PFD group. Rats except in the non-treat group were intratracheally instilled with SiO2 (25 mg/ml) or normal saline. The rats in normal saline + PFD group and the SiO2 + PFD group were given PFD (50 mg/kg) orally the next day after instillation and throughout the study. Rats were respectively sacrificed 7, 21, 42 days after instillation. The pathology changes were evaluated by Haematoxylin-eosin (HE), Van Gieson and Foot staining, and the hydroxyproline (HYP) content of pulmonary tissue was determined.</p><p><b>RESULTS</b>compared with the SiO2 group, PFD could relieve the fibrotic changes in the lungs of rats. The fibrotic degree in silicotic lesions of lungs was lower in the SiO2 + PFD group than that of SiO2 group. The HYP content in the lungs of the SiO2 + PFD group [(0.75 ± 0.12) mg/g] was significantly lower than that of the SiO2 group [(1.19 ± 0.17) mg/g] at 42 days after instillation (P < 0.05).</p><p><b>CONCLUSION</b>these data support that PFD has an antifibrotic effect against SiO2 induced lung fibrosis in rats, Which appears to be changing collagen accumulation and inhibiting pulmonary fibrosis.</p>

Abnormal liver function associated with polymorphism of GSTT1, GSTM1 and CYP2E1 in workers exposed to N, N-dimethylformamide / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate abnormal liver function associated with polymorphism of GSTT1, GSTM1 and CYP2E1 in workers exposed to N, N-dimethylformamide.</p><p><b>METHODS</b>Sixty-nine workers with abnormal liver function in a synthetic leather factory were recruited as case. One hundred and twenty five control subjects with similar work tasks were selected from the same factory. Genotypes for GSTT1 and GSTM1 were determined by multiplex PCR, and for CYP2E1 PstI by PCR-RFLP assay.</p><p><b>RESULTS</b>The frequency of positive GSTM1 was 59.42% in cases and 38.40% in control, with an odds ratio (OR) of 2.34,95% CI: 1.29-4.29 (P=0.005). For GSTT1 and CYP2E1 PstI, the frequencies of genotypes showed no significant difference between case and control.</p><p><b>CONCLUSION</b>GSTM1 positive genotype may be genetic risk factors for development of abnormal liver function in workers exposed to N, N-dimethylformamide.</p>

Chrysotile fibers induced apoptosis of human bronchial epithelial cells and effect of oxidation / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis rate and the reactive oxygen species (ROS) level induced by chrysotile fibers in BEAS-2B cells and the blockage effect of free radical scavengers on the induction of chrysotile fibers.</p><p><b>METHODS</b>The cell survival rate, the morphological variation of BEAS-2B cells, the apoptosis rate, the expression levels of gene caspase-3 and the ROS generation level were measured by using trypan blue phagocytosis, hematoxylin and eosin staining, oligonucleosomal DNA fragmentation assay, FCM, RT-PCR and fluorescent probe DCFH-DA in the suspension (0, 5, 10, 20, 100 and 200 microg/cm(2)) and the filtrate (0, 100, 200, 400, 800 and 1600 microg/ml) of chrysotile fibers. Addition of free radical scavengers such as catalase, dimethyl sulfoxide and mannitol prevented the radical generation and gene expression.</p><p><b>RESULTS</b>Survival rates of BEAS2B cells treated by the suspension (0, 5 and 10 microg/cm(2)) and the filtrate (0, 100 and 200 microg/ml) of chrysotile fibers for 24 hours were above 90%. The apoptotic rates of BEAS-2B were increased with the concentration of suspension and filtrate from chrysotile fibers (P < 0.05). Otherwise, caspase-3 mRNA and ROS were stimulated by chrysotile fibers. Free radical scavengers such as CAT, DMSO and mannitol could reduce these stimulations. The ROS blocking rate of suspension of chrysotile fibers was 23.7%, 21.6% and 11.2% respectively, and that of filtrate was 37.9%, 40.3% and 10.6% respectively.</p><p><b>CONCLUSION</b>Apoptosis is induced in BEAS-2B cells exposed to chrysotile fibers suspension and filtrate. Generation of ROS plays an important role in chrysotile fibers-induced BEAS-2B cell apoptosis.</p>

Abnormal liver function associated with occupational exposure to N, N-dimethylformamide in synthetic leathers works / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the hepatotoxic effects of N, N-dimethylformamide (DMF) in the workers of a synthetic leathers factory, and the effects on liver function of covariates such as alcohol consumption and other factors.</p><p><b>METHODS</b>The workers were classified into three groups (low, high and the control) by the concentration of DMF in workplace which was determined in the past two years. A questionnaire was drawn up for relevant demographic characteristics and other factors influencing liver function. The bloods were collected for laboratory test which included parameters especially relevant to the liver (ALT AST and gamma GT).</p><p><b>RESULTS</b>Low and high-exposure groups were significantly associated with elevated ALT and gamma GT, and high-exposure group was significantly associated with elevated Liver index. Modeling by stepwise regression analysis demonstrated that high concentration of DMF and BMI were associated with and elevated ALT, gamma GT and Liver index, besides DMF and BMI, the elevation of ALT was also associated with high TRIG. AST was only associated with alcohol consumption. The AST/ALT ration < 1 was present in 86.7% of the exposure workers of liver function abnormal.</p><p><b>CONCLUSION</b>DMF can cause liver function alternations even if air concentration of DMF was below PC-TWA. Besides the levels of DMF exposure, obesity (BMI) and alcohol consumption are covariates alternating liver function. Liver index can be a parameter for assessment liver function, and the AST/ALT ration < 1 may serve as markers of risk in health screening programs.</p>

[Clinical study of the effect of verapamil-procaine compound in prevention and treatment of acute respiratory distress syndrome after high risk operation].

OBJECTIVE: To observe the effect of verapamil-procaine compound (V-P) on prevention and treatment of acute respiratory distress syndrome (ARDS) subsequent to high risk operation. METHODS: Altogether 150 cases of major operations with high risk of ARDS were enrolled for study. They were randomly divided into three groups. V-P group: 5% glucose 500 ml and procaine 1 250 mg and verapamil 10 mg; procaine group: 5% glucose 500 ml and procaine 1 250 mg; control group: only glucose was given. The injection speed of the three groups were the same, and it was kept at 0.5 ml x h(-1) x kg(-1). The dosages of verapamil and procaine in V-P group and procaine group were doubled when the diagnosis of acute lung injury (ALI) or ARDS was confirmed. UT4000F was used in monitoring (non-invasive) blood pressure (BP), electrocardiogram (ECG), pulse oxygen saturation (SpO(2)), respiratory rate, and temperature. Blood routine and arterial blood gases measurements were intermittently performed. Diagnosis of systemic inflammatory response syndrome (SIRS), ALI and ARDS was made respectively according to published diagnostic criteria. SIRS score and acute physiology and chronic health evaluation II (APACHEII) score were performed. RESULTS: Eleven cases in V-P group, 26 in procaine group, and 42 in control group manifested symptoms and signs of SIRS. There were notable differences among groups (all P<0.01). Four patients in V-P group, 7 in procaine group, and 19 in control group were shown to develop ALI. Significant difference was found between control and V-P or procaine group (both P<0.01), but no significant difference was found between procaine group and V-P group. Twelve cases were complicated with ARDS in control group 2 weeks after the operation, and among them 5 died of multiple organ failure. There was significant difference between control group and V-P group or procaine group (both P<0.01). Two patients were complicated with acute renal failure in V-P group, 2 in procaine group, and 5 in control group. CONCLUSION: The V-P can interrupt SIRS to develop ALI, then ARDS and multiple organ dysfunction syndrome(MODS), and thus prevents and cures ARDS.