The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse tran...The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.展开更多
The method of immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep was established using monoclonal antibody. Genomic DNA was isolated from Chinese Short-tailed Hart sheep blood. U...The method of immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep was established using monoclonal antibody. Genomic DNA was isolated from Chinese Short-tailed Hart sheep blood. Using the polymerase chain reaction technique, PrP27-30 gene sequence was amplified from Chinese Short-tailed Han sheep genomic DNA. By recombinant DNA technology, the recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained. Then, using standard methodology of myeloma cell fusion, a panel of monoclonal antibodies was generated. With mAbs, scrapie in Chinese Short-tailed Han sheep was detected by immunohistochemistry assay. The recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained and a panel of six hybridoma cell lines secreting specific antibodies to Chinese Short-tailed Han sheep PrP27-30 related to scrapie was obtained with one fusion between myeloma Sp2/0 and spleen ceils from mice immunized with the purified recombinant protein. Four hybridoma cell lines can be used in immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep. So that the special monoclonal antibody developed in author＇s institute can be used to detect PrP^sc of scrapie in Chinese Short-tailed Han sheep by immunohistochemistry in China.展开更多
[ Objective] The paper was to investigate the relationship between pathological lesion and transcription, expression and distribution of heat shock protein 60 （HSP60） in kidney of chickens infected by Marek＇s disea...[ Objective] The paper was to investigate the relationship between pathological lesion and transcription, expression and distribution of heat shock protein 60 （HSP60） in kidney of chickens infected by Marek＇s disease virus （MDV）. [ Method] Tumor animal model was successfully established with chickens infected by MDV. Real-time RT-PCR, histopathological and immunohistochemistrical methods were used to detect the pathological lesion, transcription level, expression level and distribution of HSP60 in the kidney. [ Result] Obvious pathological changes appeared in kidney tissue of chicken after injection of MDV for 21 d. HSP60 was mainly distributed in the cytoplasm of tumor cell and interstitial macrophages. The expression of HSP60 in challenge group was always higher than blank control group and vaccine control group, and the difference reached extremely significant level from 28 to 86 day-ages （ P 〈 0.01 ） ; the expression of HSP60 in challenge group at 28 day-age was about 8.608 and 12.752 times of blank control group and vaccine control group, respectively. The mRNA transcription levels of HSP60 in challenge group showed a downward parabola during 7 to 42 day-ages; the mRNA transcription level of HSP60 in challenge group was rising in early stage （7 -21 d）, and then gradually dropped; it reached the peak value at 21 d, being 1.222 and 1.179 times of blank control group and vaccine control group, respectively. The mRNA transcription level of HSP60 was higher than two control groups throughout the entire experiment, and the difference reached extremely significant level from 14 to 28 day-agas （P 〈0.01 ）. [ Conclusion] HSP60 expression in kidney tissue and its mRNA transcription level can reflect MDV infection, tumor formation and development progress, with close relationship with pathogenesis of tumor diseases, which is expected to be used as one of the determination indicators for diagnosis and morbidity process of tumor diseases in chicken.展开更多
基金This work was financially supported by the National Natural Science Foundation of China （30170682, 30571400） the Specialized Research Fund for the Doctoral Program of Higher Education, China （20050307008）.
文摘The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.
基金the National Natural Sci-ence Foundation of China (C02030606)948 Project from Agriculture Ministry of China (2001-366)
文摘The method of immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep was established using monoclonal antibody. Genomic DNA was isolated from Chinese Short-tailed Hart sheep blood. Using the polymerase chain reaction technique, PrP27-30 gene sequence was amplified from Chinese Short-tailed Han sheep genomic DNA. By recombinant DNA technology, the recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained. Then, using standard methodology of myeloma cell fusion, a panel of monoclonal antibodies was generated. With mAbs, scrapie in Chinese Short-tailed Han sheep was detected by immunohistochemistry assay. The recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained and a panel of six hybridoma cell lines secreting specific antibodies to Chinese Short-tailed Han sheep PrP27-30 related to scrapie was obtained with one fusion between myeloma Sp2/0 and spleen ceils from mice immunized with the purified recombinant protein. Four hybridoma cell lines can be used in immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep. So that the special monoclonal antibody developed in author＇s institute can be used to detect PrP^sc of scrapie in Chinese Short-tailed Han sheep by immunohistochemistry in China.
基金Supported by National Natural Science Foundation of China(31101787)Natural Science Foundation of Shandong Province(ZR2010CM035)
文摘[ Objective] The paper was to investigate the relationship between pathological lesion and transcription, expression and distribution of heat shock protein 60 （HSP60） in kidney of chickens infected by Marek＇s disease virus （MDV）. [ Method] Tumor animal model was successfully established with chickens infected by MDV. Real-time RT-PCR, histopathological and immunohistochemistrical methods were used to detect the pathological lesion, transcription level, expression level and distribution of HSP60 in the kidney. [ Result] Obvious pathological changes appeared in kidney tissue of chicken after injection of MDV for 21 d. HSP60 was mainly distributed in the cytoplasm of tumor cell and interstitial macrophages. The expression of HSP60 in challenge group was always higher than blank control group and vaccine control group, and the difference reached extremely significant level from 28 to 86 day-ages （ P 〈 0.01 ） ; the expression of HSP60 in challenge group at 28 day-age was about 8.608 and 12.752 times of blank control group and vaccine control group, respectively. The mRNA transcription levels of HSP60 in challenge group showed a downward parabola during 7 to 42 day-ages; the mRNA transcription level of HSP60 in challenge group was rising in early stage （7 -21 d）, and then gradually dropped; it reached the peak value at 21 d, being 1.222 and 1.179 times of blank control group and vaccine control group, respectively. The mRNA transcription level of HSP60 was higher than two control groups throughout the entire experiment, and the difference reached extremely significant level from 14 to 28 day-agas （P 〈0.01 ）. [ Conclusion] HSP60 expression in kidney tissue and its mRNA transcription level can reflect MDV infection, tumor formation and development progress, with close relationship with pathogenesis of tumor diseases, which is expected to be used as one of the determination indicators for diagnosis and morbidity process of tumor diseases in chicken.
