In 2014, the Ministry of Health clearly pointed out the development direction of establishing a national medical consortium (reduced into the "medical association"). This paper aims to analyze the current si...In 2014, the Ministry of Health clearly pointed out the development direction of establishing a national medical consortium (reduced into the "medical association"). This paper aims to analyze the current situation of the construction of medical association in different systems in the primary medical and health service system, explore its operation mode, summarize the experience, meet the multi-level and diversified construction of medical resources through the implementation of medical association in different systems, and increase the capacity and efficiency of medical service in China.展开更多
Bovine fetal oviduct epithelial cells were transfected with constructed double marker selective vector(pCE-EGFP-IRES-Neo-dNdB) containing the enhanced green fluorescent protein (EGFP) and neomycin-resistant(Neo^r) gen...Bovine fetal oviduct epithelial cells were transfected with constructed double marker selective vector(pCE-EGFP-IRES-Neo-dNdB) containing the enhanced green fluorescent protein (EGFP) and neomycin-resistant(Neo^r) genes by electroporation, and a transgenic cell line was obtained. Somatic cell nuclear transfer (SCNT) was cartied out using the transgenic cells as nuclei donor. A total of 424 SCNT embryos were reconstructed and 208 (49.1%) of them developed to blastocyst stage. 17 blastocysts on D 7 after reconstruction were transferred to 17 surrogate calves,and 5 (29.4%) recipients were found to be pregnant. Three of them maintained to term and delivered three cloned calves.PCR and Southern blot analysis confirmed the integration of transgene in all of the three cloned calves. In addition, expression of EGFP was detected in biopsy isolated from the transgenic cloned calves and fibroblasts derived from the biopsy. Our results suggest that transgenic calves could be efficiently produced by SCNT using transgenic cells as nuclei donor. Furthermore, all cloned animals could be ensured to be transgenic by efficiently pre-screening transgenic cells and SCNT embryos using the constructed double marker selective vector.展开更多
Human lysozyme is a 130-aa (amino acid) alka-line polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ul...Human lysozyme is a 130-aa (amino acid) alka-line polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal of improving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomicDNA sequence of the human lysozme gene has been gener-ated for the first time. From 83 mice generated by microin-jection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene inlines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic mice was 0.2 mg/mL. The antibacterial activity of the whey fortransgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse wasvery low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides astrong basis for further studies into the possible application of rHlys express in mammary gland.展开更多
文摘In 2014, the Ministry of Health clearly pointed out the development direction of establishing a national medical consortium (reduced into the "medical association"). This paper aims to analyze the current situation of the construction of medical association in different systems in the primary medical and health service system, explore its operation mode, summarize the experience, meet the multi-level and diversified construction of medical resources through the implementation of medical association in different systems, and increase the capacity and efficiency of medical service in China.
文摘Bovine fetal oviduct epithelial cells were transfected with constructed double marker selective vector(pCE-EGFP-IRES-Neo-dNdB) containing the enhanced green fluorescent protein (EGFP) and neomycin-resistant(Neo^r) genes by electroporation, and a transgenic cell line was obtained. Somatic cell nuclear transfer (SCNT) was cartied out using the transgenic cells as nuclei donor. A total of 424 SCNT embryos were reconstructed and 208 (49.1%) of them developed to blastocyst stage. 17 blastocysts on D 7 after reconstruction were transferred to 17 surrogate calves,and 5 (29.4%) recipients were found to be pregnant. Three of them maintained to term and delivered three cloned calves.PCR and Southern blot analysis confirmed the integration of transgene in all of the three cloned calves. In addition, expression of EGFP was detected in biopsy isolated from the transgenic cloned calves and fibroblasts derived from the biopsy. Our results suggest that transgenic calves could be efficiently produced by SCNT using transgenic cells as nuclei donor. Furthermore, all cloned animals could be ensured to be transgenic by efficiently pre-screening transgenic cells and SCNT embryos using the constructed double marker selective vector.
基金supported by the State High-tech Research Development Program(Grant No.2002AA206111)
文摘Human lysozyme is a 130-aa (amino acid) alka-line polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal of improving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomicDNA sequence of the human lysozme gene has been gener-ated for the first time. From 83 mice generated by microin-jection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene inlines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic mice was 0.2 mg/mL. The antibacterial activity of the whey fortransgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse wasvery low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides astrong basis for further studies into the possible application of rHlys express in mammary gland.
