两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

The expression and meaning of TGF-β and TGIF in endometrosis

Objective To explore the role of TGF-βand TGIF in the pathogenesis of endometriosis. Methods The expression of TGF-β and TGIF was detected by immunohistochemistry method in the ectopic and eutopic endometrium of 30 cases with endometriosis (ec-topic endometrium group and eutopic endometrium group) and in the normal endometrium of 40 cases without endometriosis (control group). Result The expression of TGF -β in ectopic endometrium group was significantly higher than that in eutopic endometrium group and control group (P < 0.05). There was no significant difference in the expression of TGF- βbetween eutopic endometrium group and control group(P > 0.05). The expression of TGIF in ectopic endometrium group was significantly lower than that in eutopic endometrium group and control group( P <0.05). There was no significant difference in the expression of TGIF between eutopic endometrium group and control group(P > 0.05). There were negative correlation between the expressions of TGF - β and TGIF in ectopic endometrium group and eutopic endome-trium group(rs= - 0.769, - 0.549, P < 0.05). Conclusion The abnormal expression of TGF-β and TGIF in ectopic endometrium of pa-tients with endometriosis may be associated with the genesis and progression of endometriosis.     

PPAR-γ在COPD所致的肺动脉高压患者中的表达变化

Objective To investigate the relationship among peroxisome proliferators - activated receptor gamma (PPAR-γ), pul-monary arterial systolic pressure(PASP) ,and insulin resistance in Chronic Obstructive Pulmonary Disease (COPD) patients. Methods A-mong 63 COPD patients, 30 patients with level of PASP above 40mmHg were enrolled in PAH group and other 33 patients were enrolled in COPD group. Twenty healthy medical examination subjects were enrolled in control group. The expression of PPAR-γ, mRNA was detected by real time fluorescent quantitative RT- PCR. Radioimmunoassay was used to measure the level of fasting plasma insulin (FIN). Fasting plas-ma glucose (FPG) was detected by glucose oxidase method. Results The expression of PPAR-γ mRNA was significantly decreased in PAH and COPD group, while FPG, FIN and IRI increased significantly. PAH group had more increased PASP, decreased expression of PPAR-γ and higher IRI than COPD group. Expression of PPAR-γ was negatively related to PASP and IRI. Conclusions The significantly down reg-ulated expression of PPAR-γ maybe explain the higher FPG and PASP.     

Effects of silicotic alveolar macrophages exposed to SiO2 on the expression of type Ⅲ collagen and type Ⅲ procollagen in human lung fibroblasts

Objective 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-β1 antibody Methods 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-β1antibody (10μg/ml); (5) control group plus anti-TGF-β1 antibody (10 μg/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. Results 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-β1 antibody group were significantly lower than those of control plus anti-TGF-β1 antibody group (P＜0.05 or P＜0.01).Conclusion AMs exposed to SiO2 can induce the elevated pC Ⅲ and C Ⅲ expression levels in HELF by TGF-β 1 to some extent.     

足月新生儿出生体重与母婴脂联素水平的相关性研究

Objective To study the characteristics of plasma adiponectin levels exhibiting in gestation women and newborns, as well as the relationships between adiponectin levels and fetal birth weight. Methods Totally 98 subjects have been considered in this study, in-cluding venous blood samples of 36 healthy non-pregnant women (control group), 31 uncomplicated pregnant women (pregnancies group) and the cord blood samples taken at delivery of their singleton infants born at term (newborns group). The concentrations of adiponectin and insulin were determined by radioimmunoassay technique. And SPSS was used for statistical analyses. Results The adiponectin level in women of late period of gestation (WLPG) (11.10±5.72)g/ml was lower than that of other two groups, and the newborns average cord plasma level of adiponectin was (30.71±12.77) g/ml significantly higher than that of the control group (16.52±6.87) g/ml. The adipone-ctin level for WLPG was correlated with FINS and HOMA-IR (r = -0.411, -0.393, P <0.05). The adiponectin level in cord plasma was positively correlated with birth weight(r = 0.416, P < 0.05). The adiponectin level in WLPG was correlated neither with the adiponec-tin level nor body weights of their newborns. Conclusions The average cord plasma adiponectin level of newboms is significantly higher than that of the control group and WLPG. It is positively correlated with birth weight, which suggests that adiponectin may be involved in reg-ulating fetal growth. The adiponectin level in WLPG is correlated neither with the adiponectin level nor body weights of their newborns.     

兴奋性氨基酸递质系统在乐果诱导星形胶质细胞活化中的作用

Objective To study the involvement of excitatory amino acid system in astrocytes activation caused by dimethoate. Methods Pure-cultured astrocytes were gained by three passages from primary cultured rat nerve cells, then treated with 10-6,10-5,10-4 mol/L dimethoate for 48 h, 50 μmol/L and 100μmol/L MK801, a NMDA receptor blocker, was used to intervene the effects induced by 10-4 mol/L dimethoate.HPLC-FLD was utilized to measure the concentrations of excitatory amino acid (EAA), RT-PCR was used to detect the expression levels of NR2B, GLT-1, GLAST, GFAP and S100β mRNA, and immunofluresence staining method was applied to measure the expression levels of GFAP and S100β proteins. Results The expression levels of GLAST mRNA in all exposure groups were 67.8% ,68.6% and 76.2% of control level,respectively, which were significantly lower than that of control group (P＜0.05); The concentrations of EAA significantly decreased in 10-4 mol/L dimethoate group, as compared with control group (P＜0.01); the expression levels of GFAP mRNA in 10-4 mol/L dimethoate group, of S100β mRNA in 10-5 mol/L dimethoate group, of GFAP protein in 10-4 mol/L and 10-5 mol/L dimethoate groups and S100β protein in 10-4 mol/L dimethoate group were significantly higher than those in control group (P＜0.01). The expression levels of GLT-1 and GLAST mRNA in 10-4 mol/L dimethoate plus 50 μ mol/L or 100 μ mol/L MK801 groups increased significantly, as compared with 10-4 mol/L dimethoate group (P＜0.01), the expression levels of NR2B mRNA in 10-4 mol/L dimethoate plus 50 μ mol/L or 100 μmol/L MK801 groups increased significantly, as compared with control group (P＜0.05 or P＜0.01); the concentration of Glu in 10-4 mol/L dimethoate plus 100 μ mol/L MK801 group increased significantly, as compared with 10-4 mol/L dimethoate group (P＜0.01); the expression levels of GFAP mRNA and protein in10-4 mol/L dimethoate plus 50 μ mol/L or 100 μ mol/L MK801 groups decreased significantly (P＜0.01); S100β protein expression level in 50 μ mol/L MK801 intervention group was significantly higher than thatl in control group (P＜0.01). Conclusion Excitatory amino acid system involved in astrocytes activation caused by dimethoate. MK801 was useful to control astrocytes gliosis.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.     

两种膨润土细胞毒性的体外试验研究

Objective To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.Methods The cytotoxicity of two kinds of bentonite was detected using CCK8 assay,neutral red uptake(NRU) assay,lactate dehydrogenase(LDH) leakage assay,apoptosis assay and hemolysis assay.In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0,0.3125,0.6250,1.2500 and 2.5000 mg/ml for ten min.In other four assays,human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0,10,20,30,60,120 and 180 μg/ml for four h.Results In hemolysis assay,the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P＜0.05);in CCK-8 assay,the cellular activities in acid bentonite group at the doses ≥30 μg/ml and in organic bentonite group at the doses ≥20μg/ml were significantly lower than that of control (P＜0.01);the similar results appeared in NRU assay and LDH assay,and the dose-effect relationship was observed in above 4 assays.In apeptosis assay,the early apoptosis cell rates in acid bentonite group at the dose of 180 μg/ml and in organic bentonite group at the doses of 120,180 μg/ml were significantly higher than that of control (P＜0.05).Moreover,the results of five in vitro assays indicated the eytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.Conclusion Two kinds of bentonite could induce cytotoxicity,such as apoptosis and damage of cell membrane.The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.