The nucleotide sequence of the melanocortin-l-receptor （MC1R） gene was studied with the help of the polymerase chain reaction （PCR）, in which the protein structure in Chinese Holstein was predicted, and the molecu...The nucleotide sequence of the melanocortin-l-receptor （MC1R） gene was studied with the help of the polymerase chain reaction （PCR）, in which the protein structure in Chinese Holstein was predicted, and the molecular mechanism of the red coat color was investigated. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was performed to genotype the individuals. The bioinformatics and biotechnology softwares were used to predict the secondary structure of MC1R. The results showed that the EE genotype was the dominant genotype in Chinese Holstein Black and White herd, whereas, it was ee in Chinese Holstein Red and White herd. The secondary structure of the mutational MC1R protein was changed and the deletion mutation caused an earlier termination in translation, which led to the formation of the red coat color. The allele E was mainly associated with the black coat color, whereas, e was associated with red.展开更多
Background Enhanced apoptosis of cytotrophoblasts in early pregnancy is associated with high risk of intrauterine growth retardation and preeclampsia, which are two common pregnant complications. Its etiological facto...Background Enhanced apoptosis of cytotrophoblasts in early pregnancy is associated with high risk of intrauterine growth retardation and preeclampsia, which are two common pregnant complications. Its etiological factors remain unclear. Cytotrophoblasts share some traits with innate immune cells and may show response to lipopolysaccharide. This study was conducted to demonstrate whether lipopolysaccharide has apoptosis-inducing effects on cytotrophoblast and the role of innate immune reaction in this process. Methods Cytotrophoblasts were isolated from eady pregnant villous tissues and cultured with serum-free medium. Subsequently, cytotrophoblasts were treated with lipopolysaccharide at the concentrations of 0 （control）, 25, 50, 100 and 200 ng/ml for 24 hours. Apoptosis of cytotrophoblasts was determined by light microscopy, Hoechst 33258 DNA staining with a fluorescent microscope, transmission electron microscope and annexin V-fluorescein isothiocyanate-cenjugated / propidium iodide （PI） staining with flow cytometry. Then expression of caspase-3 was detected by Western blot. Confocal immunofluorescence technique was used to detect tumor necrosis factor a expression in cytotrophoblasts. The levels of tumor necrosis factor a in the culture medium were detected by enzyme-linked immunosorbent assay. Results Under light, fluorescence microscope and transmission electron microscope, characteristic altemations of apoptosis in cytotrophoblasts were observed after lipopolysaccharide treatment. Flow cytometry results showed that lipopolysaccharide significantly increased apoptosis indexes of cytotrophoblasts. Significant statistical differences were found in the above groups （P≤0.01）. The mean relative densities of bands corresponding to caspase-3 were significantly increased in groups treated with lipopolysaccharide, as compared with the normal control （P〈0.001）. Tumor necrosis factor a expression was found to increase in cytotrophoblasts by confocal immunofluorescence technique and in culture medium by enzyme-linked immunosorbent assay after lipopolysaccharide treatment. A positive correlation was found between tumor necrosis factor a expression and apoptosis indexes of cytotrophoblasts （r=0.747, P〈0.001）. Conclusion Apoptosis of cytotrophoblasts could be induced by lipopolysaccharide, in which innate immune reaction is the important mechanism.展开更多
The title compound phenyl(6-phenyl-3-p-tolyl-6,7-dihydro-5 H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazin-7-yl)methanone(PTM),designed using 4-amino-3-p-tolyl-1H-1,2,4-triazole-5(4 H)-thione as the starting material,was su...The title compound phenyl(6-phenyl-3-p-tolyl-6,7-dihydro-5 H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazin-7-yl)methanone(PTM),designed using 4-amino-3-p-tolyl-1H-1,2,4-triazole-5(4 H)-thione as the starting material,was successfully synthesized via a two-step synthesis route and finally characterized by NMR,FT-IR,single-crystal X-ray diffraction and mass spectrometry techniques.The crystal structure of trans-PTM was obtained from X-ray diffraction:C24H20N4OS,Mr=412.52,monoclinic system,space group P21/c,a=16.650(3),b=13.876(3),c=8.812(2)A,β=100.340(3)°,V=2002.8(7)A^3,F(000)=865,Z=4,Dc=1.3680 g/cm^3,λ=0.71073A,μ=0.186 mm^-1 and the final R=0.0786 for 3514 unique reflections with 2044 observed ones(Ⅰ>2σ(Ⅰ)).The biological activities of antimicrobial and regulation of plant growth of PTM were investigated.The results indicated that PTM showed weak antimicrobial activity on Bacillus pumilus,while promoted the growth of radish and inhibited that of wheat in a dose-dependent manner.Therefore,PTM may be developed as a potential drug to promote the growth of dicotyledonous plants or as an herbicide to inhibit that of monocotyledonous plants in the future.展开更多
基金National 863 Program of China(2006AA1021D9,2007AA10Z169)High Technology Independent Innovation Project from Shandong Academy of Agricultural Science s(2006YCX028)Youth Science Foundation from Shandong Academy of Agricultural Sciences of China(2006YQN038)
文摘The nucleotide sequence of the melanocortin-l-receptor （MC1R） gene was studied with the help of the polymerase chain reaction （PCR）, in which the protein structure in Chinese Holstein was predicted, and the molecular mechanism of the red coat color was investigated. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was performed to genotype the individuals. The bioinformatics and biotechnology softwares were used to predict the secondary structure of MC1R. The results showed that the EE genotype was the dominant genotype in Chinese Holstein Black and White herd, whereas, it was ee in Chinese Holstein Red and White herd. The secondary structure of the mutational MC1R protein was changed and the deletion mutation caused an earlier termination in translation, which led to the formation of the red coat color. The allele E was mainly associated with the black coat color, whereas, e was associated with red.
文摘Background Enhanced apoptosis of cytotrophoblasts in early pregnancy is associated with high risk of intrauterine growth retardation and preeclampsia, which are two common pregnant complications. Its etiological factors remain unclear. Cytotrophoblasts share some traits with innate immune cells and may show response to lipopolysaccharide. This study was conducted to demonstrate whether lipopolysaccharide has apoptosis-inducing effects on cytotrophoblast and the role of innate immune reaction in this process. Methods Cytotrophoblasts were isolated from eady pregnant villous tissues and cultured with serum-free medium. Subsequently, cytotrophoblasts were treated with lipopolysaccharide at the concentrations of 0 （control）, 25, 50, 100 and 200 ng/ml for 24 hours. Apoptosis of cytotrophoblasts was determined by light microscopy, Hoechst 33258 DNA staining with a fluorescent microscope, transmission electron microscope and annexin V-fluorescein isothiocyanate-cenjugated / propidium iodide （PI） staining with flow cytometry. Then expression of caspase-3 was detected by Western blot. Confocal immunofluorescence technique was used to detect tumor necrosis factor a expression in cytotrophoblasts. The levels of tumor necrosis factor a in the culture medium were detected by enzyme-linked immunosorbent assay. Results Under light, fluorescence microscope and transmission electron microscope, characteristic altemations of apoptosis in cytotrophoblasts were observed after lipopolysaccharide treatment. Flow cytometry results showed that lipopolysaccharide significantly increased apoptosis indexes of cytotrophoblasts. Significant statistical differences were found in the above groups （P≤0.01）. The mean relative densities of bands corresponding to caspase-3 were significantly increased in groups treated with lipopolysaccharide, as compared with the normal control （P〈0.001）. Tumor necrosis factor a expression was found to increase in cytotrophoblasts by confocal immunofluorescence technique and in culture medium by enzyme-linked immunosorbent assay after lipopolysaccharide treatment. A positive correlation was found between tumor necrosis factor a expression and apoptosis indexes of cytotrophoblasts （r=0.747, P〈0.001）. Conclusion Apoptosis of cytotrophoblasts could be induced by lipopolysaccharide, in which innate immune reaction is the important mechanism.
基金supported by the grants from Zhangzhou Natural Science Foundation of China(No.ZZ2018J39)The work reported in this manuscript has been granted a Chinese Patent(No.ZL201710632570.6)。
文摘The title compound phenyl(6-phenyl-3-p-tolyl-6,7-dihydro-5 H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazin-7-yl)methanone(PTM),designed using 4-amino-3-p-tolyl-1H-1,2,4-triazole-5(4 H)-thione as the starting material,was successfully synthesized via a two-step synthesis route and finally characterized by NMR,FT-IR,single-crystal X-ray diffraction and mass spectrometry techniques.The crystal structure of trans-PTM was obtained from X-ray diffraction:C24H20N4OS,Mr=412.52,monoclinic system,space group P21/c,a=16.650(3),b=13.876(3),c=8.812(2)A,β=100.340(3)°,V=2002.8(7)A^3,F(000)=865,Z=4,Dc=1.3680 g/cm^3,λ=0.71073A,μ=0.186 mm^-1 and the final R=0.0786 for 3514 unique reflections with 2044 observed ones(Ⅰ>2σ(Ⅰ)).The biological activities of antimicrobial and regulation of plant growth of PTM were investigated.The results indicated that PTM showed weak antimicrobial activity on Bacillus pumilus,while promoted the growth of radish and inhibited that of wheat in a dose-dependent manner.Therefore,PTM may be developed as a potential drug to promote the growth of dicotyledonous plants or as an herbicide to inhibit that of monocotyledonous plants in the future.
