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Objective To analyze the case characteristics of invited suicidal death in order to provide references for field investigation and forensic identification.Methods Analyze the scene conditions,the sex,ages,relationship,ways of meeting,suicidal incentives,and the ways of committing suicide of the casualties by collecting 15 cases of invited suicidal death in Minhang and Hongkou District of Shanghai from 2006 to 2016.Results In these 15 cases,male more than female,most of the casualties had unambiguous incentives and they were youth from age 18 to 44,and invited suicide cases via internet is significantly increased,most of them were committing charcoal-burning suicide and they chose to commit suicide in hotels and inns mostly.The manners of suicide are mostly coincident.Conclusions The characteristics of the cases mentioned above help the forensic experts to catch the point of the scene investigation in similar cases and make quick and accurate judgments on the manner and the characteristics of deaths.
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<p><b>OBJECTIVE</b>To screen the differentially expressing genes between silicotic lung tissue and normal lung tissue, to identify the differentially expressing genes of matrix metalloproteinase-12 (MMP-12) and Cathepsin E and to explore the roles of those genes in silicosis development.</p><p><b>METHODS</b>Thirty male SD rats were divided randomly into two groups: control group (6 rats) and exposure group (24 rats) which was exposed to SiO2 by intra-tracheal perfusion. On the 30 th, 60 th and 90 th days after exposure, 8 rats in model group and 2 rats in control group were executed and the lung tissues were obtained. The morphologic changes of lung tissues were observed with HE staining and VG staining under a light microscope. The gene microarrays were used to identify differentially expressing genes of lung tissues in rats exposed to SiO2 for 60 days. Two significantly up-regulated genes, MMP-12 and Cathepsin E, were validated using RT-PCR, immunohistochemistry and Western Blot assay.</p><p><b>RESULTS</b>A total of 338 differentially expressing genes were identified from the 26 962 genes between silicotic rats and normal rats, including 267 up-regulated genes and 71 down-regulated genes. The results of RT-PCR showed that in the lung tissues of exposure group on the 30 th, 60 th and 90 th days, the mRNA expression levels of MMP-12 were 4.306, 5.338, 6.713 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.434, 2.974, 3.889 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the mRNA expression levels of MMP-12 were 1.435, 1.746, 2.069 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.372, 1.663, 2.103 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the expression levels of MMP-12 protein were 1.214, 1.531, 1.959 times higher than those in the control group, the expression levels of Cathepsin E protein were 1.262, 1.828, 1.907 times higher than those in the control group, respectively. Compared with the control group, the mRNA and protein expression levels of MMP-12 and Cathepsin E in lung tissues of exposure group were significantly up-regulated (P < 0.05).</p><p><b>CONCLUSION</b>The differentially expressing genes in rat lung tissues screened by gene chip were validated, which suggested that a complex gene regulatory network may be contributed to occurrence of silicosis. MMP-12 and Cathepsin E genes may be involved in the development of silicotic pulmonary fibrosis by degrading the basement membrane of alveolar wall and participating in the immune response.</p>
Subject(s)
Animals , Male , Rats , Cathepsin E , Genetics , Metabolism , Gene Expression , Lung , Metabolism , Matrix Metalloproteinase 12 , Genetics , Metabolism , Rats, Sprague-Dawley , Silicosis , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect and regulation of extracellular signal-regulated kinase (ERK1/2) signaling pathway on the expression of transforming growth factor-beta1 (TGF-beta1) in human embryonic lung fibroblasts induced by SiO2.</p><p><b>METHODS</b>Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and in the presence or absence of SiO2 (50 ug/ml) exposition for 18h, and then the conditioned supernatants were used to incubate HELF. The expressions of TGF-beta1, of the HELF acted with the conditioned AM supernatant fluid were detected with the immunocytochemistry method after treatment with PD98059 of inhibitor of ERK.</p><p><b>RESULTS</b>The expression of TGF-beta1 in HELF of the SiO2 treatment group (OD value is 0.322 7 +/- 0.023 8) exceed blank group (OD value is 0.163 7 +/- 0.019 6) and AM control group (OD value is 0.240 6 +/- 0.022 5) by the immunocytochemistry method. But the expression of TGF-beta1 had reduction in some extent in the PD98059 intervention group (OD value is 0.271 1 +/- 0.022 9). The values were statistically different (P < 0.05).</p><p><b>CONCLUSION</b>ERK inhibitor PD98059 have inhibition effect on the expression of transforming growth factor-beta1 and expression of cytokine of human embryonic lung fibroblasts stimulated by SiO2. The study indicate that the proliferation and collagen production of HELF activated by SiO2 are mediated by ERK/MAPK signal pathway in some extent. PD98059 may antagonizes silica-induced lung fibrosis by inhibiting the expression of transforming growth factor-beta1.</p>
Subject(s)
Humans , Male , Middle Aged , Bronchoalveolar Lavage Fluid , Cell Biology , Cells, Cultured , Fibroblasts , Metabolism , Flavonoids , Pharmacology , Lung , Cell Biology , Metabolism , MAP Kinase Signaling System , Macrophages, Alveolar , Cell Biology , Metabolism , Silicon Dioxide , Silicosis , Metabolism , Transforming Growth Factor beta1 , MetabolismABSTRACT
<p><b>UNLABELLED</b>Effects of silicotic alveolar macrophages exposed to SiO2 on the expression of type III collagen and type</p><p><b>OBJECTIVE</b>To study the effects of supernatant of alveolar macrophages (AM) exposed to SiO2 on the expression of type III collagen and type III procollagen in human lung fibroblasts (HELF) and the intervention effects of anti-TGF-beta1 antibody.</p><p><b>METHODS</b>AMs collected from a silicotic by bronchoalveolar lavage were divided into 2 parts, one part was exposed to SiO2 and other part served as control. The supernatant was obtained from AMs cultured for 18 h. HELF were divided into (1) exposure group, which was added with supernatant from AMs exposed to SiO2; (2) control group, which was added with the supernatant from AMs not exposed to SiO2; (3) blank control group, which was added with DMEM; (4) exposure group plus anti-TGF-beta1 antibody (10 microg/ml); (5) control group plus anti-TGF-beta1 antibody (10 microg/ml). (1)-(3) groups were cultured for 6, 12, 18, 24, 36, 48h, respectively. (4)-(5) groups were cultured for 18, 24, 36, respectively. Immunocytochemical test and Western blot assay were used to detect pC III expression levels in HELF and C III expression levels in the supernatant of HELF culture, respectively.</p><p><b>RESULTS</b>The pC III expression levels of exposure group were 0.1423 +/- 0.0107, 0.1624 +/- 0.0011, 0.1925 +/- 0.0050, 0.2421 +/- 0.0097 and 0.2103 +/- 0.0103, respectively, which were significantly higher than those (0.1212 +/- 0.0079, 0.1414 +/- 0.0058, 0.1620 +/- 0.0081, 0.1965 +/- 0.0103, 0.1715 +/- 0.0116) of control group (P < 0.05 or P < 0.01). The C III levels of exposure group were (0.2559 +/- 0.0061, 0.3249 +/- 0.0110, 0.4171 +/- 0.0193, 0.5441 +/- 0.0452, 0.4751 +/- 0.0252), respectively, which were significantly higher than control group (0.2296 +/- 0.0121, 0.2778 +/- 0.0116, 0.3367 +/- 0.0269, 0.3722 +/- 0.0214). The pC III and C III expression levels of exposure plus anti-TGF-beta1 antibody group were significantly lower than those of control plus anti-TGF-beta1 antibody group (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>AMs exposed to SiO2 can induce the elevated pC IIII and C III expression levels in HELF by TGFbetaP1 to some extent.</p>
Subject(s)
Humans , Male , Middle Aged , Cells, Cultured , Collagen Type III , Metabolism , Fibroblasts , Metabolism , Lung , Cell Biology , Macrophages, Alveolar , Cell Biology , Metabolism , Silicon Dioxide , Toxicity , Silicosis , Metabolism , Transforming Growth Factor beta , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of ERK1/2 signal pathway activated by SiO₂ in the proliferation of human embryonic lung fibroblast mediated by silicotic alveolar macrophages.</p><p><b>METHOD</b>The alveolar macrophages (AM) harvested from silicotic sufferers by bronchoalveolar lavage (BAL) were interacted with SiO₂ suspension once more. HELF, pretreated with the inhibitor PD98059 (50 µmol/L) for 1 hour, were stimulated by conditional supernatant fluid of silicotic sufferers. The experimentation have been classificated four group: blank group, AM control group, SiO₂ treatment group, PD98059 intervention group. The proliferation activity and expressions of Phospho-ERK1/2 of lung fibroblast activated by AM supernatant fluids of silicotic are detected with the MTT assay, flow cytometry and Western blot method after being pretreatmented with PD98059.</p><p><b>RESULT</b>The A values of cell proliferation in SiO₂ treatment group and AM control group are 2.6 and 2.0 times that of blank group, in which the difference was statistically significant (P < 0.05). Comparing with SiO₂ treatment group, the A values of every concentrations of PD98059 intervention group decreased with a dose-response relationship, after 10, 25 and 50 µmol/L PD98059 intervention. The 25 and 50 µmol/L PD98059 intervention group were 72.1% and 48.5% of SiO₂ treatment group, which the difference is statistic (P < 0.05). The expression of phospho-ERK1/2 in SiO₂ treatment group was up, which appeared in 15 min and apparent activated in 30 min (A value is 0.4653 ± 0.0265), and then still in the higher state afterwards declined after 60 min. In addition to 15 min, the expression of phospho-ERK1/2 protein in SiO₂ treatment group at each time point are 1.25, 1.23, 1.25 times over the same period AM control group respectively, the differences were statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>The silicotic supernatant of alveolar macrophages have promote proliferation of HELF and activation of ERK1/2, which may involve in the development of silicosis pathogenesis by ERK1/2 signal pathway.</p>
Subject(s)
Humans , Male , Middle Aged , Cell Proliferation , Cells, Cultured , Culture Media, Conditioned , Fibroblasts , Metabolism , Flavonoids , Pharmacology , Lung , Cell Biology , Macrophages, Alveolar , Metabolism , Mitogen-Activated Protein Kinase 1 , Metabolism , Mitogen-Activated Protein Kinase 3 , Metabolism , Signal Transduction , Silicon Dioxide , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the proliferation effect of the AM supernatant incubated activation of p38 mitogen activated protein kinases (p38MAPK) signal transduction pathway in human embryonic lung fibroblasts, and to participate in the development of fibrosis in silicosis.</p><p><b>METHODS</b>The silicotic alveolar macrophages were collected by bronchoalveolar lavage and incubated in vitro in the DMEM medium containing SiO₂ (50 µg/ml) and DMEM medium without SiO₂ for 18 h. Then the AM supernatant incubated for 18 h was collected. HELFs were isolated by organize paste block method, and incubated with AM supernatants. HELFs were divided into four groups: blank control groups, AM groups, SiO₂ + AM groups, SB203580 + SiO₂ + AM groups. The proliferation in the HELF was detected with MTT method and Flow cytometry.</p><p><b>RESULTS</b>The proliferation in the HELF acted with the conditioned AM supernatant fluid were more than blank control groups, AM groups and SB203580 + SiO₂ + AM groups [average optical density: (0.48 ± 0.03) vs (0.29 ± 0.01), (0.38 ± 0.02), (0.33 ± 0.03)], the values with MTT method were statistically different (P < 0.05); Proliferous index with flow cytometry in SiO₂ + AM groups (18.12 ± 0.82) was bigger than blank control groups (9.24 ± 0.48), AM groups (14.76 ± 0.43) and SB203580 + SiO₂ + AM groups (11.71 ± 0.70) and the values were statistically different(P < 0.05).</p><p><b>CONCLUSIONS</b>The AM supernatant stimulated by silicon dioxide can accelerate the proliferation in the HELF by activation of p38MAPK signal transduction pathway.</p>
Subject(s)
Adult , Humans , Male , Cell Proliferation , Cells, Cultured , Culture Media, Conditioned , Fibroblasts , Cell Biology , Pathology , Lung , Cell Biology , MAP Kinase Signaling System , Macrophages, Alveolar , Cell Biology , Signal Transduction , Silicon Dioxide , Pharmacology , Silicosis , Metabolism , Pathology , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effect of SiO(2) on the expression of platelet derived growth factor (PDGF) in human silicotic alveolar macrophages (AM) and human embryonic lung fibroblasts (HELF).</p><p><b>METHODS</b>Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to SiO(2) for 3, 6, 12, 18, 24 and 36 h. The cultured supernatant at 24 h was incubated with human embryonic lung fibroblasts for 6, 12, 18, 24, 36 and 48 h. The immunocytochemistry and Western blot were used to detect the level of expression of PDGF in lung fibroblasts and their supernatant respectively. (3)H-proline was used to detect the synthesis and secretion of collagen in HELF.</p><p><b>RESULTS</b>The expression of the PDGF in the supernatant of alveolar macrophages exposed to SiO(2) increased significantly and reached the peak at 24 h (average optical density: 0.282 +/- 0.019 vs 0.214 +/- 0.014, P < 0.01) with ELISA. The expression of PDGF in lung fibroblasts and their supernatant increased at different time (6, 12, 18, 24, 36 and 48 h) with immunocytochemistry and Western blot respectively when incubated with the cultured supernatant of silicotic AM exposed to SiO(2). The expression of PDGF was significantly different from the control group (P < 0.05). The synthesis and secretion of collagen in FB were increased markedly when incubated with the cultured supernatant of AM stimulated by SiO(2) compared with the control group.</p><p><b>CONCLUSION</b>SiO(2) may affect the expression of PDGF and synthesis of collagen through AM mediation and participate in the formation of lung fibrosis.</p>
Subject(s)
Humans , Male , Middle Aged , Cells, Cultured , Collagen , Metabolism , Fibroblasts , Metabolism , Macrophages, Alveolar , Metabolism , Platelet-Derived Growth Factor , Metabolism , Silicon Dioxide , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the effect of culture supernatant of alveolar macrophage alveolar macrophages (AM) stimulated by SiO2 on the expression of matrix metalloproteinases (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagen of fibroblast human embryonic lung fibroblasts (HELF) in the development of silicosis fibrosis.</p><p><b>METHODS</b>AMs were collected from a silicotic patient by bronchoalveolar lavage and exposed to SiO2, cultured human embryo lung fibroblast were allocated into a treated group, a control group, a positive group, and a blank group. HELF was incubated with the cultured supernatant of AMs for 6, 12, 18, 24, 36, 48 h. Immunocytochemical and Western blot technology were used to detect MMP-1 and TIMP-1 expressions in HELF and collagen expression in supernatant of HELF respectively.</p><p><b>RESULTS</b>The supernatant of AM exposed to SiO2 significantly decreased the expressions of MMP-1 (0.0605 +/- 0.0201, 0.0519 +/- 0.0117, 0.0412 +/- 0.0105 and 0.0213 +/- 0.0106 in the treated group at 18, 24, 36 and 48 h) compared with the control group and the blank group (P < 0.05, P < 0.01) but stimulated expressions of TIMP-1 and collagen (P < 0.05, P < 0.01). The ratio of TIMP-1 to MMP-1 increased. The ratio of TIMP-1 to MMP-1 was positively correlated with the expression of collagen III (r = 0.88, P < 0.01).</p><p><b>CONCLUSION</b>Through AM mediation SiO2 can accelerate the expression of TIMP-1 and collagen, and inhibit the expression of MMP-1. The imbalance between the expression of TIMP-1 and that of MMP-1 is related with the abnormal increase in collagen III.</p>
Subject(s)
Humans , Male , Middle Aged , Cells, Cultured , Collagen Type III , Metabolism , Fibroblasts , Metabolism , Macrophages, Alveolar , Matrix Metalloproteinase 1 , Metabolism , Silicon Dioxide , Toxicity , Silicosis , Pathology , Tissue Inhibitor of Metalloproteinase-1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in human silicotic alveolar macrophages (AM).</p><p><b>METHODS</b>Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3 h, 6 h, 12 h, 18 h, 24 h and 36 h. The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.</p><p><b>RESULTS</b>The expressions of MMP-9 in the AM increased clearly along with the time, reached peak at 24 h when detected with zymography (average optical density: 3.061+/-0.153 vs 2.851+/-0.164, P<0.05); and reached peak at 18h when detected with immunological method (average optical density: 0.386+/-0.037 vs 0.322+/-0.034, P<0.05). The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P>0.05).</p><p><b>CONCLUSION</b>SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.</p>
Subject(s)
Humans , Cells, Cultured , Macrophages, Alveolar , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Silicon Dioxide , Toxicity , Silicosis , Pathology , Tissue Inhibitor of Metalloproteinase-1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the expression of KiSS-1, nuclear factor kappa B (NF-kappaB) p50 and matrix metalloproteinase 9 (MMP-9) in breast cancer tissue and the relationship with clinicpathological factors.</p><p><b>METHODS</b>Immunohistochemical staining for KiSS-1, NF-KappaBp50, and MMP-9 protein was performed in 152 cases of human breast tissue [92 cases of BC, 30 cases of epithelial hyperplasia, and 30 cases of peritumoral breast tissue (PMT)] and 54 cases of axillary lymph node metastases. In-situ hybridization for KiSS-1 mRNA was done in 50 cases of breast cancer, and 20 cases of PMT.</p><p><b>RESULTS</b>(1) The expression of KiSS-1 gene was significantly higher in well-differentiated breast cancer than in PMT, and this expression progressively decreased with decreasing degree of tumor differentiation, increasing pathological grade, TNM stage and the presence of lymph node metastases. The expression of KiSS-1 gene in lymph node metastasis was markedly lower than the corresponding primary tumor. There was correlation between the expression of KiSS-1 mRNA and KiSS-1 protein in breast cancer group. (2) The expression of NF-kappaKBp50 and MMP-9 increased progressively with decreasing degree of tumor differentiation, increasing TNM stage, large tumor size ( >2 cm) and the presence of lymph node metastases.</p><p><b>CONCLUSIONS</b>The expression of KiSS-1 protein showed negative correlation with that of NF-kappaBp50 and MMP-9 respectively. MMP-9 protein expression was positively correlated with NF-kappap50 protein expression. These suggest that the genes of KiSS-1, NF-kappaBp50 and MMP-9 could be involved in the progression and metastasis of breast cancer.</p>
Subject(s)
Female , Humans , Breast Neoplasms , Metabolism , Pathology , Gene Expression Regulation, Neoplastic , Kisspeptins , Lymphatic Metastasis , Matrix Metalloproteinase 9 , Genetics , Metabolism , NF-kappa B , Genetics , Metabolism , RNA, Messenger , Metabolism , Statistics as Topic , Tumor Suppressor Proteins , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effect of the cultured supernatant of human silicotic alveolar macrophages (AM) on the expression of the collagen type I in human embryonic lung fibroblasts.</p><p><b>METHODS</b>Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 18 h. Then the cultured supernatant were used to culture human embryonic lung fibroblasts for 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 72 h. Then detected collagen anabolism and secretion with (3)H-proline detected the expression of the procollagen type I in the fibroblast with immunological method detected the quantity of collagen Type I in FB supernatant with Western blot.</p><p><b>RESULTS</b>The anabolism and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO(2), Along with the time, the expression of collagen type I increased. In cultured supernatant of silicotic AM exposed to SiO(2), ((3)H-proline: 1096.500 +/- 76.400, 707.000 +/- 62.160, OD: 0.314 +/- 0.011, OD: 14.218 +/- 0.342.</p><p><b>CONCLUSION</b>SiO(2) may affect the expression of collagen through AM mediation and participate in the formation of lung fibrosis.</p>