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Objective To investigate the relationship between Helicobacter pylori (H. pylori) infection and concentration of reactive oxygen species (ROS) in AGS cells. Methods AGS cells were cultured with either Hp11638 (CagA~+ , VacA~+ ) extract or Hp11638 mutant (CagA~+ , VacA~-) extract for 48 hours, then the cells and supernatants were collected. The concentration of ROS in AGS cells was measured by flow cytometry. The eytochrome C reduction was detected by spectrophotometer at 550 nm. Results The ROS levels in the AGS cells were correlated with two H. pylori strains in a concentration- and time-dependent manner. The ROS levels in AGS cells treated with Hp11638 extract in different concentrations or times were correspondingly higher than those treated with Hp11638 mutant extract. Similar results were found in examination of cytochrome C reduction. Conclusion The elevation of ROS in AGS cells is related to effects of H. pylori proteins, and the VaeA protein involves in the process.
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Objective To study the effects of lipopolysaccharide(LPS) on the expression of toll like receptor 4 (TLR4), reactive oxygen species(ROS) and on the proliferation of cells as well as secretion of six proinflammmatory cytokines including TNF-α, IL-1, IL-6, IL-8, IL-10, IL-12 levels in SKOV3 cells. And to explore the mechanism of SKOV3 cells in regulation. Methods Cultured primary SKOV3 cells were stimulated with different concentrations of LPS (0.01 μg/ml, 0.1 μg/ml, 1 μg/ml, 10 μg/ml and 20 μg/ml) for 4 h, the TLR4 expression in SKOV3 cells were examined by flow cytometry;1 μg/ml LPS stimulated SKOV3 for 4 h, 8 h, 12 h, 24 h respectively, the TLR4 expression and cell cycle in SKOV3, cell proliferation, ROS level as well as cells and TNF-α and IL-1, IL-6, IL-8, IL-10, IL-12 levels in the culture medium were assayed by flow cytometry, MTT, CBA assay respectively. Results LPS with different concentrations of LPS stimulation in-duced an increased TLR4 expression, however, the expression was reduced when LPS concentration up to 10 μg/ml. LPS stimulation for 4 h, 8 h induced an increased TLR4 expression and cell proliferation. Stimulated for 24 h, however, the TLR4 expression and cell growth were inhibited in S period. Meanwhile, LPS stimulation for 4 h, 8 h, 12 h, 24 h induced a higher ROS secretion in comparison with control group. LPS stimulation induced a stronger cytokine response in comparison with control group, as demonstrated by the production of TNF-α, IL-1, IL-6, IL-8 secretion in cultured SKOV3 cells, while IL-10 and IL-12 with low expression have no obvious difference in the all medium samples. Conclusion TLR4 expression, cell proliferation, ROS and proin-flammmatory cytokine secretion could be induced in SKOV3 through LPS stimulation. The study provide new ex-periment evidences for human ovarian cells SKOV3 immunity regulation and inflammation reaction to promote cells inhibition after LPS stimulation.
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Objective To explore the impact of acute Toxoplasma gondii infection on cerebral proteins and nerve growth in mice by 2D electrophoresis. Methods The cerebral proteins from C57BL/6J mice infected with Toxoplasma gondii and normal paired mice were extracted. The discrepant proteins were checked by 2D electrophoresis. Isoelectric focusing was determined as the first direction (immobilized pH gradient gel 3-10) ,SDS-PAGE as the second direction to execute 2D electrophoresis, and PDQuest 1.0 software was used to analyze 2D electrophoretogram. Results The protein spots in Toxoplasma gondii infected mice and normal paired mice were (132 ±10) and (170 ± 13) , respectively. After the analysis by PDQuest 1. 0 software, only 19 protein spots were found to express in infected mice and only 37 protein spots were found to express in normal paired mice. Additionally, the obvious quantitative changes in a part of proteins of the cerebrum in the both group occurred. Conclusion There are obvious changes in cerebral proteins from mice with acute Toxoplasma gondii infection, which provids useful clues for studying the cerebral proteins injury in acute Toxoplasma gondii infected mice and the new cure drug.
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To investigate the effect of Toxoplasma gondii infection upon the expression of brain-derived neurotrophic factor (BDNF) mRNA and N-methyl-D-aspirate receptor (NMDA) subunits NR2A and NR2B,Wistar rats of 4 weeks old were randomly divided into 3 groups with 10 rats in each group in which 2 mL suspensions of T.gondii tachyzoits in the concentrations of 2×10~7/mL and 2×10~5/mL were injected intra-peritoneally to rats in group A and group B respectively, serving as the experimental groups, while 2 mL of sterile physiologic saline was injected intra-peritoneally in group C serving as the control group. Four weeks after injection, the expressions of BDNF mRNA and BDNF protein in the brain tissues were detected by in situ hybridization and immunohistochemical assay and the expressions of NR2A and NR2B immune activity in the hippocampal CA1,CA3 and DG were investigated by using computer-assisted image analysis system. Compared with the control group, the expression of BDNF protein in the hippocampus of the experimental groups was significantly enhanced [(64.27±23.18), (50.39±19.34) vs (44.68±22.74)/mm~2,P＜0.05]. In addition, the increased expressions of BDNF mRNA in the hippocampus of the experimental groups were also demonstrated [(0.13±0.02), (0.12±0.02) vs (0.09±0.01); P＜0.05]. In the expression of the NR2A protein, their expressions in group A and B of rats were significantly lower than that of group C in CA3 (P＜0.05),but there was no significant change in CA1 and DG. In the expression of NR2B protein, the expressions in group A and B were also lower than that of group C in CA1 and CA3, and had no significant change in DG. It is evident that the expressions of BDNF mRNA and BDNF protein in hippocampal tissues were significantly increased following chronic infection with T.gondii, supporting the hypothesis that BDNF may be involved in the intrinsic neuro-protective mechanism.
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Objective To search for a biomarker from the serum of Schistosoma japonicum infected rabbits for early diagnosis of schistosomiasis. Methods The sera were obtained from different periods of the infected rabbits. The serum proteins were generated by WCX kit (Bruker Daltonics GmBH) and analyzed by the technique of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Results In the mass range from 1 000 to 12 000 Da, sixty-three proteins were captured by WCX kit. ClinProTools software was used to find the differential expressed proteins. The result revealed 7 distinct proteins compared with normal serum. Among them,1 787 Da protein expression was increased (P