A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium （MEM） supplemented with fetal bovine serum （FBS...A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium （MEM） supplemented with fetal bovine serum （FBS） and 10 ng ml4 basic fibroblast growth factor （bFGF）. Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number （2n=46）. The cells were successfully transfected with green fluorescent protein （GFP） reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus （LCDV） and found to be susceptible to the virus in cytopathic effect （CPE） observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.展开更多
A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 ...A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 passages for over 200 d in DMEM medium supplemented with 15% FBS （fetal bovine serum）, 2.5 ng/cm^3 bFGF （basic fibroblast growth factor）, 1 ng/cm^3 LIF （leukemia inhibitory factor） and 50 mmol/dm^3 2-ME （2-mecaptoethanol）. The cells grew well in the temperature range of 24-30 ℃ and the optimal growth temperature was 24 ℃. FBS and bFGF concentrations are the two key components for CSEC cells proliferation. Chromosome analysis reveals that CSEC cells have a normal diploid karyotype with 2n=42t. The significant fluorescent signals were observed in CSEC after transfection with the GFP reporter gene, suggesting that the CSEC cell line can be used as a useful tool for transgenic and genetic manipulation studies. CSEC cells showed the cytopathic effect （CPE） after infection with lymphosystis disease virus （LCDV） in 2 d. Moreover, the LCDV particles can be observed in the cytoplasm of virus-infected cells by electron microscopy. It suggests that CSEC could be potentially used for the study of aquatic virus.展开更多
2016年8月24日19:00至25日08:00(北京时)在500 h Pa副热带高压控制下,甘肃中东部、陕西关中出现强对流暴雨,19个乡镇出现大暴雨,最大降水量达158.7 mm,小时最大降水量达79.1 mm,且伴随雷电天气,呈现典型的强对流天气特征。利用卫星、雷...2016年8月24日19:00至25日08:00(北京时)在500 h Pa副热带高压控制下,甘肃中东部、陕西关中出现强对流暴雨,19个乡镇出现大暴雨,最大降水量达158.7 mm,小时最大降水量达79.1 mm,且伴随雷电天气,呈现典型的强对流天气特征。利用卫星、雷达、地面加密资料、ECMWF细网格资料、NCEP再分析资料和常规观测资料,重点分析造成短时强降水的中尺度系统的发生、发展以及中尺度对流系统传播特征。结果表明,大暴雨主要由2个中尺度对流系统产生;中尺度对流系统的发生、发展与中尺度地面辐合线有密切关系;低层动力场的切变扰动在六盘山两侧形成东西向的辐合线,雷暴单体在辐合线附近强烈发展,整体随辐合线向偏南方向移动;六盘山以西的辐合线影响甘肃中部,六盘山东侧的辐合线在移动中分裂为两段:东段继续向偏南方向移动,影响甘肃陇东;西段移到关中西部时,沿峡谷进入关中西部的偏北风,受地形影响转为西北风,使辐合线由东西向转为南北向,雷暴单体随辐合线沿地形走向自西向东移动,影响关中;有利的动力、热力因素,对六盘山东侧中尺度对流系统的加强和传播方向的改变起到重要作用。展开更多
基金supported by grants from State 863High-Technology Rand Project of China(2006AA09Z406,2006AA10A401)Taishan Scholar Project of Shan-dong Province
文摘A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium （MEM） supplemented with fetal bovine serum （FBS） and 10 ng ml4 basic fibroblast growth factor （bFGF）. Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number （2n=46）. The cells were successfully transfected with green fluorescent protein （GFP） reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus （LCDV） and found to be susceptible to the virus in cytopathic effect （CPE） observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.
基金The State "863" High-Technology Research and Development Project of China under contract Nos 2006AA09Z406 and 2006AA10A401the National Natural Science Foundation of China under contract No. 40376047the Taishan Scholar Project of Shandong Province and MOA for returned scientists from abroad
文摘A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 passages for over 200 d in DMEM medium supplemented with 15% FBS （fetal bovine serum）, 2.5 ng/cm^3 bFGF （basic fibroblast growth factor）, 1 ng/cm^3 LIF （leukemia inhibitory factor） and 50 mmol/dm^3 2-ME （2-mecaptoethanol）. The cells grew well in the temperature range of 24-30 ℃ and the optimal growth temperature was 24 ℃. FBS and bFGF concentrations are the two key components for CSEC cells proliferation. Chromosome analysis reveals that CSEC cells have a normal diploid karyotype with 2n=42t. The significant fluorescent signals were observed in CSEC after transfection with the GFP reporter gene, suggesting that the CSEC cell line can be used as a useful tool for transgenic and genetic manipulation studies. CSEC cells showed the cytopathic effect （CPE） after infection with lymphosystis disease virus （LCDV） in 2 d. Moreover, the LCDV particles can be observed in the cytoplasm of virus-infected cells by electron microscopy. It suggests that CSEC could be potentially used for the study of aquatic virus.
文摘2016年8月24日19:00至25日08:00(北京时)在500 h Pa副热带高压控制下,甘肃中东部、陕西关中出现强对流暴雨,19个乡镇出现大暴雨,最大降水量达158.7 mm,小时最大降水量达79.1 mm,且伴随雷电天气,呈现典型的强对流天气特征。利用卫星、雷达、地面加密资料、ECMWF细网格资料、NCEP再分析资料和常规观测资料,重点分析造成短时强降水的中尺度系统的发生、发展以及中尺度对流系统传播特征。结果表明,大暴雨主要由2个中尺度对流系统产生;中尺度对流系统的发生、发展与中尺度地面辐合线有密切关系;低层动力场的切变扰动在六盘山两侧形成东西向的辐合线,雷暴单体在辐合线附近强烈发展,整体随辐合线向偏南方向移动;六盘山以西的辐合线影响甘肃中部,六盘山东侧的辐合线在移动中分裂为两段:东段继续向偏南方向移动,影响甘肃陇东;西段移到关中西部时,沿峡谷进入关中西部的偏北风,受地形影响转为西北风,使辐合线由东西向转为南北向,雷暴单体随辐合线沿地形走向自西向东移动,影响关中;有利的动力、热力因素,对六盘山东侧中尺度对流系统的加强和传播方向的改变起到重要作用。
