In order to create the Male Sterile Line in tobacco, the anti-sense fragment of HSP70 gene was linked to anther specific expression promoter TA29 and the reconstructed vector was transformed into tobacco by Agrobacter...In order to create the Male Sterile Line in tobacco, the anti-sense fragment of HSP70 gene was linked to anther specific expression promoter TA29 and the reconstructed vector was transformed into tobacco by Agrobacterium mediated transformation, and the transformants were then screened. Gus and spot blotting hybridization analysis of the transformants indicated that anti-sense fragment of HSF70 gene had been integrated into tobacco genome and expressed, thus the male sterile tobacco line was obtained. Microscope observation of anther and pollen showed that pistils of transgenic tobacco were normal, whereas anthers and pollens were fairly abortive in the same transgenic tobacco flower, comparing with pistils and stamens in control plants. The ratio of HSI：＇70 protein before and after heat shock in mitochondrial was found to be 1.39 in control tobacco plants and 1.01 in transgenic tobacco sterile lines. This is suggested that the anti-sense gene fragment of HSP70 can effectively inhibit the expression of HSP70 protein and lead to transgenic male sterility in tobacco flowers. The assay provided a new genetic engineering method for male sterility creation in plants.展开更多
AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70...AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70 and grp94 in 78 human esophageai cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis. RESULTS: Both esophageai cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageai carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8% (56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageai cancer and adjacent normal tissues (P<0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P<0.01). CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70 is related to the differentiation of esophageai carcinoma.展开更多
AIM： To investigate the ultra-structural changes and heat shock protein 70 （HSP70） expression in the pancreas of rats with endotoxic shock and to detect their possible relationship. METHODS： A total of 33 Wistar r...AIM： To investigate the ultra-structural changes and heat shock protein 70 （HSP70） expression in the pancreas of rats with endotoxic shock and to detect their possible relationship. METHODS： A total of 33 Wistar rats were randomly divided into three groups： control group （given normal saline）, small dose lipopolysaccharide （LPS） group （given LPS 5 mg/kg） and large dose LPS group （given LPS 10 mg/kg）. Pancreas was explanted to detect the ultrastructural changes by TEM and the HSP70 expression by immunohistochemistry and Western blot. RESULTS： Rats given small doses of LPS showed swelling and loss of mitochondrial cristae of acinar cells and increased number of autophagic vacuoles in the cytoplasm of acinar cells. Rats given large doses of LPS showed swelling, vacuolization, and obvious myeloid changes of mitochondrial cristae of acinar cells, increased number of autophagic vacuoles in the cytoplasm of acinar cells. HSPT0 expression was increased compared to the control group （P〈0.05）. CONCLUSION： Small doses of LPS may induce stronger expression of HSP70, promote autophagocytosis and ameliorate ultra-structural injuries.展开更多
Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of Hsp...Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of Hsp70 gene family using ClustalW algorithm. The authors used an in silico approach to find a homology between more than one accession numbers of DNA sequences, X67711.2 was for Oryza sativa Hsp70, AY372071.1 was for Nicotiana tabacum Hsp70 and L41253.2 was for Lycopersicon esculentum Hsc70. The three accession numbers which were retrieved by the BLASTn program depend on their expected value （E-value）. Multiple sequence alignment was performed by ClustalW algorithm to produce a conserved blocks and determine the consensus region which had been used to produce the forward and reverse primer by the primer select module of DNAStar Lasergene V7 and In-Silco PCR module of FASTPCR program ver.4.0.8 was performed to detect the melting temperatures （Tm） and predict the PCR product size, The results of designed degenerate primer showed that there was a homology found between the designed primers and the DNA templates for the three accession numbers with at least 80% identity. The result of degenerate PCR showed that the three bands of the amplified PCR products of the three accession numbers were detected at the same molecular weight of marker （400 bp） with a difference about 15 pb compared to the in silco PCR product （385 pb）. In conclusion, this study focused on the importance of using the clustalW algorithm for designing the degenerate primer.展开更多
Objective： To investigate the protective effects of acuptmcture pretreatment on ischemic myocardium, the protective mechanism of acupuncture pretreatment on ischemic myocardium was explored by observing the cardiac m...Objective： To investigate the protective effects of acuptmcture pretreatment on ischemic myocardium, the protective mechanism of acupuncture pretreatment on ischemic myocardium was explored by observing the cardiac muscle cell apoptosis and the expression of HSP70 mRNA of ischemia-reperfusion injury rats treated with acupuncture pretreatment. Methods： Sixty-four Wistar rats were randomly divided into eight groups： control group, sham surgery group, ischemia-reperfusion group, ischemia pretreatment group, manual acupuncture pretreatment group （once a day）, electroacupuncture pretreatment group （once a day）, manual acupuncture pretreatment group （twice a day）, and electroacupuncture pretreatment group （twice a day）. The reperfusion model of rat myocardial ischemia was made. Expression of HSP70 mRNA was assayed by in situ hybridization, and cell apoptosis by TUNEL. Results： Compared with those in the control group and the sham surgery group, the apoptosis and the expression of HSP70 mRNA were increased in the ischemia-reperfusion group. Compared with those in the ischemia-reperfusion group, the cardiac muscle cell apoptosis was decreased and the HSP70 mRNA was increased in the rats treated with acupuncture pretreatment; meanwhile, acupuncture pretreatment twice a day had stronger effects than acupuncture pretreatment once a day and ischemia pretreatrnent. Conclusion： Acupuncture pretreatment can inhibit the cardiac muscle cell apoptosis, and up-regulate the expression of HSP70 mRNA in ischemia-reperfusion rats. Acupuncture pretreatment twice a day has stronger effects than pretreatment once a day.展开更多
Objective： To observe the effect of preventively used moxibustion on the expression of apoptosis-related factors in rats with acute gastric mucosal damage （GMD）, and analyze the relationship between those factors a...Objective： To observe the effect of preventively used moxibustion on the expression of apoptosis-related factors in rats with acute gastric mucosal damage （GMD）, and analyze the relationship between those factors and heat shock protein 70 （HSP70）, for discovering the mechanism of moxibustion for protecting gastric mucosa from the aspect of mitochondria signal transduction of apoptosis. Methods： Thirty-two Wistar rats were randomly divided into binding, model, acupoint-moxibustion and non-acupoint-moxibustion groups, 8 in each group. Moxibustion was applied preventively to acupoint-moxibustion and non-acupointmoxibustion groups respectively for 8 d. Rats in binding and model groups were served with restraint without moxibustion. The GMD models were induced in groups except for A group, and the expression of cytochrome c （Cyt-c） in gastric mucosa was measured by Western-blot analysis, the expression of HSP70, cell apoptosis index （AI）, apoptotic protease activating factor 1 （Apaf-1）, Caspase-9, Caspase-3, Bcl-2 and Bax in gastric mucosal cells were studied by immunohistochemical detection. Results： Compared with binding group, the expressions of HSP70, AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax of gastric mucosa in model group were significantly higher （P〈0.05 or P〈0.01）. Compared with model group, the expressions of HSP70 and Bcl-2 were significantly higher （P〈0.01）, while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly lower （P〈0.01） in acupoint-moxibustion group.Compared with the acupoint-moxibustion group, the expressions of HSP70 and Bcl-2 were significantly lower （P〈0.01）, while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly higher （P〈0.05 or P〈0.01） in non-acupoint-moxibustion group. Conclusion： Moxibustion inhibits the apoptosis of rats＇ gastric mucosal cells and protect them from damage, through up-regulating the expression of HSP70, and modulating the expressions of Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax which were related to the mitochondria signal transduction of apoptosis.展开更多
基金Supported by National Natural Science Foundation of China(30970247)Hunan Provincial Science Fund for Distinguished Young Scholars(11JJ1007)~~
文摘In order to create the Male Sterile Line in tobacco, the anti-sense fragment of HSP70 gene was linked to anther specific expression promoter TA29 and the reconstructed vector was transformed into tobacco by Agrobacterium mediated transformation, and the transformants were then screened. Gus and spot blotting hybridization analysis of the transformants indicated that anti-sense fragment of HSF70 gene had been integrated into tobacco genome and expressed, thus the male sterile tobacco line was obtained. Microscope observation of anther and pollen showed that pistils of transgenic tobacco were normal, whereas anthers and pollens were fairly abortive in the same transgenic tobacco flower, comparing with pistils and stamens in control plants. The ratio of HSI：＇70 protein before and after heat shock in mitochondrial was found to be 1.39 in control tobacco plants and 1.01 in transgenic tobacco sterile lines. This is suggested that the anti-sense gene fragment of HSP70 can effectively inhibit the expression of HSP70 protein and lead to transgenic male sterility in tobacco flowers. The assay provided a new genetic engineering method for male sterility creation in plants.
基金Supported by the Youth Research Scientific Foundation of Beijing (02-31)
文摘AIM: To investigate the expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human esophageai carcinoma and adjacent normal tissues. METHODS: The expression of HSP70 and grp94 in 78 human esophageai cancer and adjacent normal tissues was studied by immunohistochemistry and pathology photograph analysis. RESULTS: Both esophageai cancer and adjacent normal tissues could express HSP70 and grp94. Of the 78 cases of esophageai carcinoma, 95.0%(72/78) showed positive HSP70, mainly stained in nuclei, while grp94 was mainly stained in cell plasma, and the positive rate was 71.8% (56/78).There was a significant difference in the expression of HSP70 and grp94 between esophageai cancer and adjacent normal tissues (P<0.01). Compared with adjacent normal tissues, there was a significant difference between differential types and HSP70 expression (P<0.01). CONCLUSION: HSP70 and grp94 express differently in cell plasma and nuclei. The expression intensity of HSP70 is related to the differentiation of esophageai carcinoma.
基金Supported by the National Natural Science Foundation of China,No.30170849
文摘AIM： To investigate the ultra-structural changes and heat shock protein 70 （HSP70） expression in the pancreas of rats with endotoxic shock and to detect their possible relationship. METHODS： A total of 33 Wistar rats were randomly divided into three groups： control group （given normal saline）, small dose lipopolysaccharide （LPS） group （given LPS 5 mg/kg） and large dose LPS group （given LPS 10 mg/kg）. Pancreas was explanted to detect the ultrastructural changes by TEM and the HSP70 expression by immunohistochemistry and Western blot. RESULTS： Rats given small doses of LPS showed swelling and loss of mitochondrial cristae of acinar cells and increased number of autophagic vacuoles in the cytoplasm of acinar cells. Rats given large doses of LPS showed swelling, vacuolization, and obvious myeloid changes of mitochondrial cristae of acinar cells, increased number of autophagic vacuoles in the cytoplasm of acinar cells. HSPT0 expression was increased compared to the control group （P〈0.05）. CONCLUSION： Small doses of LPS may induce stronger expression of HSP70, promote autophagocytosis and ameliorate ultra-structural injuries.
文摘Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of Hsp70 gene family using ClustalW algorithm. The authors used an in silico approach to find a homology between more than one accession numbers of DNA sequences, X67711.2 was for Oryza sativa Hsp70, AY372071.1 was for Nicotiana tabacum Hsp70 and L41253.2 was for Lycopersicon esculentum Hsc70. The three accession numbers which were retrieved by the BLASTn program depend on their expected value （E-value）. Multiple sequence alignment was performed by ClustalW algorithm to produce a conserved blocks and determine the consensus region which had been used to produce the forward and reverse primer by the primer select module of DNAStar Lasergene V7 and In-Silco PCR module of FASTPCR program ver.4.0.8 was performed to detect the melting temperatures （Tm） and predict the PCR product size, The results of designed degenerate primer showed that there was a homology found between the designed primers and the DNA templates for the three accession numbers with at least 80% identity. The result of degenerate PCR showed that the three bands of the amplified PCR products of the three accession numbers were detected at the same molecular weight of marker （400 bp） with a difference about 15 pb compared to the in silco PCR product （385 pb）. In conclusion, this study focused on the importance of using the clustalW algorithm for designing the degenerate primer.
文摘Objective： To investigate the protective effects of acuptmcture pretreatment on ischemic myocardium, the protective mechanism of acupuncture pretreatment on ischemic myocardium was explored by observing the cardiac muscle cell apoptosis and the expression of HSP70 mRNA of ischemia-reperfusion injury rats treated with acupuncture pretreatment. Methods： Sixty-four Wistar rats were randomly divided into eight groups： control group, sham surgery group, ischemia-reperfusion group, ischemia pretreatment group, manual acupuncture pretreatment group （once a day）, electroacupuncture pretreatment group （once a day）, manual acupuncture pretreatment group （twice a day）, and electroacupuncture pretreatment group （twice a day）. The reperfusion model of rat myocardial ischemia was made. Expression of HSP70 mRNA was assayed by in situ hybridization, and cell apoptosis by TUNEL. Results： Compared with those in the control group and the sham surgery group, the apoptosis and the expression of HSP70 mRNA were increased in the ischemia-reperfusion group. Compared with those in the ischemia-reperfusion group, the cardiac muscle cell apoptosis was decreased and the HSP70 mRNA was increased in the rats treated with acupuncture pretreatment; meanwhile, acupuncture pretreatment twice a day had stronger effects than acupuncture pretreatment once a day and ischemia pretreatrnent. Conclusion： Acupuncture pretreatment can inhibit the cardiac muscle cell apoptosis, and up-regulate the expression of HSP70 mRNA in ischemia-reperfusion rats. Acupuncture pretreatment twice a day has stronger effects than pretreatment once a day.
文摘Objective： To observe the effect of preventively used moxibustion on the expression of apoptosis-related factors in rats with acute gastric mucosal damage （GMD）, and analyze the relationship between those factors and heat shock protein 70 （HSP70）, for discovering the mechanism of moxibustion for protecting gastric mucosa from the aspect of mitochondria signal transduction of apoptosis. Methods： Thirty-two Wistar rats were randomly divided into binding, model, acupoint-moxibustion and non-acupoint-moxibustion groups, 8 in each group. Moxibustion was applied preventively to acupoint-moxibustion and non-acupointmoxibustion groups respectively for 8 d. Rats in binding and model groups were served with restraint without moxibustion. The GMD models were induced in groups except for A group, and the expression of cytochrome c （Cyt-c） in gastric mucosa was measured by Western-blot analysis, the expression of HSP70, cell apoptosis index （AI）, apoptotic protease activating factor 1 （Apaf-1）, Caspase-9, Caspase-3, Bcl-2 and Bax in gastric mucosal cells were studied by immunohistochemical detection. Results： Compared with binding group, the expressions of HSP70, AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax of gastric mucosa in model group were significantly higher （P〈0.05 or P〈0.01）. Compared with model group, the expressions of HSP70 and Bcl-2 were significantly higher （P〈0.01）, while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly lower （P〈0.01） in acupoint-moxibustion group.Compared with the acupoint-moxibustion group, the expressions of HSP70 and Bcl-2 were significantly lower （P〈0.01）, while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly higher （P〈0.05 or P〈0.01） in non-acupoint-moxibustion group. Conclusion： Moxibustion inhibits the apoptosis of rats＇ gastric mucosal cells and protect them from damage, through up-regulating the expression of HSP70, and modulating the expressions of Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax which were related to the mitochondria signal transduction of apoptosis.
