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Heavy ions are the positively charged nuclei of the atomic number greater than two in the universe, which mainly comes from the charged particles of the solar system outside. Heavy ions occupy only 1% in cosmic ray, but because they are is capable of forming the Bragg peak, resulting in huge energy deposition, they have much more severe biological effects on immune organ than electromagnetic radiation in the cosmic rays. This paper outlines the biological effects of heavy ions on several aspects of immune organs, immune cells and chromosomes, and provide a theoretical basis for the study of the impact of heavy ion radiation on the immune system.
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Objective To investigate the effect of asarone on the zebrafish embryonic development and behavior. Methods The 3 hours post fertilization(hpf) zebrafish embryos were exposed to different concentrations(0, 25,50,100,200 and 400 μmol/L) of asarone solution. The asarone solution was replaced each 24 h. Microscope was used to observe embryos morphology at 24,48,72 and 96 hpf, respectively. Spontaneous movements at 24 hpf, heart rate at 48 hpf, hatch rate, deformity rate and mortality rate were evaluated.The sepn1 gene expression of zebrafish in 96 hpf was detected by RT-qPCR. With Noldus tracking system the behaviors of zebrafish larvae exposed to asarone were recorded. Results After zebrafish embryos were exposed to the different concentrations of asarone, their spontaneous movements were decreased. Compared with the control group, the 100,200 and 400 μmol/L groups were significantly decreased (P<0.01). There was a significant difference(P<0.01) in 48 hpf heart rate between all asarone groups and control group. Spine curvature, pericardial edema, yolk sac edema and other deformity were observed at 72 hpf and 96 hpf. Compared with the control group, the 100, 200 and 400 μmol/L groups showed significanly decreased hatch rate(P<0.01). There was a significant difference (P<0.05) in mortality rate between the 400 μmol/L group and control group. With concentrations of asarone increasing, the speed and distance of movement of 200 and 400 μmol/L group were significantly reduced compared to the control group(P<0.05). There was a significant difference between the 200,400 μmol/L groups and the control group in activity(P< 0.01).At 96 hpf, the expression of sepn1 gene in arone groups were decreased compared to control group, especially the 100 μmol/L group(P<0.05), and the 200 and 400 μmol/L groups(P<0.01). Conclusion Asarone had teratogenic effect on zebrafish embryos and larvae ,and inhibitory effect on larvae behavior .We should be careful to use asarone in infant medication.
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Objective Using silkworm expression system to express human serum paraoxonase 1 (PON1) fusion protein with protein transduction domain P11 and to study its biological activity of fighting aginst the toxicity of dichlorvos. Methods P11-PON1 fusion gene was constructed in cloning sites of silkworm pFastBac 5B vector, the vector and was transformed to silkworm DH10BmBac competent cells. Virus particles and 5 instar silkworm was infected 96 hours after infection, parasites were collected and lyophilized crushed and preserved at-80℃. The protien was dissolved, sonicated and centrifuged before used. Supernatants were harvested. The fusion protein P11-PON1 proportion of the total protein was analyzed with SDS-PAGE electrophoresis and protein content was calculated. Mouse and zebrafish models were used to evaluate P11-PON1 fusion protein bioactivity. Each mouse was treated with 1 mg P11-PON1 fusion protein via intragastric or rectal administration. 0 and 3 hours after administration, 20 mg/kg dichlorvos were injected intraperitoneally. The status of intoxication was observed and the survival rate was scored. P11-PON1 fusion protein with concentrations of 1, 2.5, 5, 10, 20 mg/L was dissolved in zebrafish breeding water respectively. 0 and 3 hours after exposing dichlorvos were added with a final concentration of 50 mg/L. Observe their behavioral change.The survival rate of zebrafish was recorded. Results The content of P11-PON1 fusion protein was 8%of silkworm total protein. In mice experiments, P11-PON1 fusion protein by intragastric adminstration did not increase the survival of mouse. By intraperitoneal injection with dichlorvos 0h after rectal adminstration with protein,the survival rate of mouse did not significantly increase. In contrast, the mouse intraperitoneally injected with dichlorvos 3h after adminstration with protein, the survival rate increased extremely significantly (P < 0.01). In zebrafish experiments, the zebrafish exposed to dichlorvos 0 h after adminstration with protein, the survival rate was not significantly improved, while exposed to dichlorvos 3h after admindtration, the survival rate significantly increased. The survival rate of 20, 10, 5 mg/L group were 62.5%, 62.5%and 50%respectively at 24 h time point, compared to the control group. The survival rate increased extremely significantly (P < 0.01). 2.5 mg/L group was 41.7%, with the survival rate increasing significantly (P < 0.05). However, the survival rate of 1 mg/L group was 16.7%, compared to the control group. The increase had no sistatistical significance. Conclusion The PTD-containing PON1 fusion protein can be expressed in silkworm. Pretreatment with the fusion protein in mice and zebrafish decreased the toxicity of dichlorvos.
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OBJECTIVE To investigate the develop mental toxicity of muscone to embryos. METHODS With zebrafish embryos as a model,The 3 h post fertilization (hpf)embryos were exposed to muscone at 5,10,20,40,80 and 160 μmol·L -1 culture solutions for 96 h and inspected daily with mi-croscopy for larval morphology.The drug solution was replaced every 24 h.Spontaneous move ments were checked at 24 hpf.Heart rate at 48 hpf,hatching rate,e mbryo deformity rate and mortality rate were evaluated.The expression of sepn1 was determined with real-ti me quantitative PCR technique at 96 hpf.RESULTS The 24 hpf spontaneous move ments showed no significant difference.At 48 hpf, spine curvature,pericardial ede ma,yolk sac ede ma,and abnormal swi mming were observed.In addition, the 48 hpf heart beats(10 s)was decreased fro m 26.5 ±1 .0 to 18.0 ±1 .9(P <0.01 ).At 48 hpf , hatching rate of 5 ~40 μmol·L -1 decreased(P <0.05),while of 160 μmol·L -1 increased (P <0.05) co mpared with muscone 0 μmol·L -1 .Muscone had little effect on hatching rate at other ti me points;Mal-formation rate and mortality rate at higher concentrations were up to 100%.The sepn1 gene expression at 96 hpf in the exposure groups decreased co mpared with that of control group(P <0.01 ).CONCLU-SION Muscone had toxic effects on the develop ment of zebrafish embryos,including spine curvature, abnormal swi mming,and pericardial ede ma.These effects may be related to the inhibition of sepn1 gene expression by muscone.
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OBJECTlVE To establish Tg(-6.3CYP3A65∶EGFP) transgenic zebrafish for quick, intuitive detection of heavy metals ( copper, cadmium and zinc) , dioxin-like PCBs ( PCB126) and other environmental pollutants. METHODS Tol2 transposon system was used to generate transgenic zebrafish lines Tg(-6.3CYP3A65∶EGFP) in which CYP3A65 promoter regualated labeled fluorescence. The effect of heavy mentals ( copper, cadmium and zinc ) and PCB126 on the relative amounts of CYP3A65 gene expression was determined by observing the change in fluorescence intensity. RESULTS The relative gene expression of CYP3A65 was significantly increased after 96 h exposure to copper 0.1 and 0.2μmol·L-1 , cadmium 0.35 and 0.7μmol·L-1 , zinc 1.5 and 3μmol·L-1 , and PCB126 2-32μmol·L-1 , respectively ( P<0.01) , but decreased after 96 h exposure to copper 0. 9 μmol·L-1 , cadmium 2. 7 and 5.4 μmol·L-1 , and zinc 24μmol·L-1 , respectively( P<0.01) . CYP3A65 gene expression was significantly increased after 168 h exposure to copper 0.1 and 0.2 μmol·L-1 , cadmium 0.35 and 0.7 μmol·L-1 , zinc 1.5 and 3 μmol·L-1, and PCB126 2-32 μmol·L-1, respectively(P<0.01), but decreased after 168 h exposure to copper 0.9 μmol·L-1, cadmium 2.7 and 5.4 μmol·L-1, and zinc 12 and 24 μmol·L-1( P<0.05) , in a concentration-dependent manner. CONCLUSlON The results suggest that zebrafish CYP3A65 gene expression and the CYP3A65 labeled fluorescence lines can be another candidate biomarker for detecting environmental pollutants.