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基于Cre-loxP重组酶系统构建肺泡Ⅱ型上皮细胞特异性敲除SENP1基因小鼠
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作者 杨坤 章容 +4 位作者 吴越 雷小平 谌云川 康兰 董文斌 《中国组织工程研究》 CAS 北大核心 2025年第14期2943-2950,共8页
背景:前期在体外成功构建了SENP1基因沉默的人肺泡上皮细胞系,在细胞水平上研究了SENP1在高氧性肺损伤中的作用。目的:基于Cre-loxP重组酶系统构建肺泡Ⅱ型上皮细胞特异性敲除SENP1基因小鼠模型。方法:将SENP1^(flox/-)小鼠自交得到SENP... 背景:前期在体外成功构建了SENP1基因沉默的人肺泡上皮细胞系,在细胞水平上研究了SENP1在高氧性肺损伤中的作用。目的:基于Cre-loxP重组酶系统构建肺泡Ⅱ型上皮细胞特异性敲除SENP1基因小鼠模型。方法:将SENP1^(flox/-)小鼠自交得到SENP1^(flox/flox)和SENP1^(flox/-)小鼠;将Sftpc-Cre^(+/+)小鼠与野生型小鼠交配获得更多的Sftpc-Cre^(+/-)小鼠。将Sftpc-Cre^(+/+)或子代Sftpc-Cre^(+/-)小鼠与SENP1^(flox/-)或子代SENP1^(flox/flox)小鼠进行杂交,获得SENP1^(flox/-)Sftpc-Cre^(+/-)双杂合小鼠。将SENP1^(flox/-)Sftpc-Cre^(+/-)小鼠与SENP1^(flox/flox)小鼠杂交,获得SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠。剪鼠尾提取基因组DNA,行PCR扩增,扩增产物经琼脂糖凝胶电泳确定小鼠基因型。取SENP1^(flox/flox)和SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠肺组织行免疫荧光双标实验及Western blot以验证SENP1敲除效果;取SENP1^(flox/flox)和SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠心、肝、肺、肾组织行苏木精-伊红染色以观察两组小鼠各脏器的组织形态。结果与结论:琼脂糖凝胶电泳正确筛选出SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠。免疫荧光双标实验显示,与SENP1^(flox/flox)小鼠相比,SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠肺组织中SENP1的平均荧光强度降低(P<0.01),且SENP1和Sftpc未见明显共定位(P<0.01)。Western blot结果显示,与SENP1^(flox/flox)小鼠相比,SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠肺组织中SENP1蛋白表达降低(P<0.001)。苏木精-伊红染色结果显示SENP1^(flox/flox)和SENP1^(flox/flox)Sftpc-Cre^(+/-)小鼠的心、肝、肺和肾脏组织形态无明显改变。该研究利用Cre-loxP重组酶系统成功构建了肺泡Ⅱ型上皮细胞特异性敲除SENP1基因小鼠,为后续研究SENP1基因在以肺泡Ⅱ型上皮细胞为主要损伤细胞的肺疾病如支气管肺发育不良、特发性肺纤维化中的作用提供了良好的工具。 展开更多
关键词 SENP1 cre-loxP重组酶系统 肺泡Ⅱ型上皮细胞 条件性基因敲除 小鼠
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The environmental implication of diatom fossils in the surface sediment of the Changjiang River estuary(CRE) and its adjacent area 被引量:3
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作者 FAN Xin CHENG Fangjin +1 位作者 YU Zhiming SONG Xiuxian 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2019年第2期552-567,共16页
In order to depict the distribution of diatom fossils in surface sediments and to establish a reliable reference data for further paleoenvironmental study in the Changjiang (Yangtze) River estuary and its adjacent wat... In order to depict the distribution of diatom fossils in surface sediments and to establish a reliable reference data for further paleoenvironmental study in the Changjiang (Yangtze) River estuary and its adjacent waters, the diatom fossils from 34 surface sediment samples and their relationship with environmental variables were analyzed by principal component analysis and redundancy correspondence analysis. The diversity and abundance of diatom fossils were analyzed. Some annual average parameters of the overlying water (salinity, temperature, turbidity, dissolved oxygen, depth, dissolved inorganic nitrogen, dissolved inorganic phosphate and dissolved inorganic silicate) were measured at each sampling site. A total of 113 diatom taxa and one silicoflagellate species were identified in the investigation area. Diatom fossils were better preserved in fine sediments. The absolute abundance of diatom fossils did not significantly diff er between inshore and off shore areas, the species diversity decreased from inshore to off shore. This may be because high nutrients and low salinity promoted the growth of more brackish species in coastal waters. The diatom taxa were divided into three groups, on the basis of their response and indication to environmental changes. For example, Actinocyclus ehrenbergii and Cyclotella stylorum were dominant in coastal waters (Group 1 and Group 3) with high nutrients and low salinity;the relative abundances of Paralia sulcata and Podosira stelliger were significantly higher in off shore sites (Group 2, average 39.5%), which were characterized by high salinity and deep water. Four environmental variables (salinity, dissolved inorganic nitrogen, temperature and water depth) explained the composition and distribution of diatom taxa independently ( P< 0.05), this finding can be applied in further paleoenvironmental reconstruction research in this area. 展开更多
关键词 DIATOM FOSSILS CHANGJIANG River estuary surface SEDIMENTS environmental variables
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应用Cre-LoxP系统构建牛结节性皮肤病病毒缺失TK基因毒株
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作者 张玉哲 任善会 +10 位作者 姚威 龚真莉 张红强 柳民意 尤婷 王相伟 李积雲 殷相平 孙跃峰 陈豪泰 万学瑞 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第10期4517-4529,共13页
利用同源重组技术及Cre-LoxP系统平台,构建牛结节性皮肤病病毒(lumpy skin disease virus,LSDV)缺失TK基因毒株,为研制出安全、高效的LSDV基因工程疫苗提供备选毒株。以TK基因为靶基因,利用Cre-LoxP系统平台,红色荧光蛋白(RFP)为筛选标... 利用同源重组技术及Cre-LoxP系统平台,构建牛结节性皮肤病病毒(lumpy skin disease virus,LSDV)缺失TK基因毒株,为研制出安全、高效的LSDV基因工程疫苗提供备选毒株。以TK基因为靶基因,利用Cre-LoxP系统平台,红色荧光蛋白(RFP)为筛选标记,采用重叠PCR方法扩增左、右同源臂以及RFP基因表达框并进行融合,将其克隆至pUC19T载体,构建基因缺失转移载体pUC19T-LSDV-ΔTK-RFP。将转移载体重组质粒转染至Vero细胞,并感染LSDV/CHA/FJ/2021毒株,构建LSDV-ΔTK-RFP重组病毒。采取蚀斑法、有限稀释法纯化LSDV-ΔTK-RFP。然后,利用Cre-LoxP系统,在MDBK-Cre细胞中从病毒基因组中切除RFP标签,采取蚀斑法和有限稀释法筛选并纯化LSDV-ΔTK毒株。利用PCR及测序方法对重组毒株进行鉴定,并测定其一步生长曲线。结果表明成功构建缺失TK基因的重组毒株(LSDV-ΔTK),重组毒株复制力略低于野生毒株。应用Cre-LoxP系统构建的缺失TK基因LSDV毒株,具有良好的遗传稳定性和复制生长特性,为后续LSDV基因工程疫苗的研制提供了候选毒株,同时拓展了Cre-LoxP系统的应用范围。 展开更多
关键词 牛结节性皮肤病病毒 TK基因 cre/LOXP系统 重组毒株
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Does reduced sediment load contribute to increased outbreaks of harmful algal blooms off the Changjiang Estuary? 被引量:2
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作者 WANG Baodong XIN Ming +2 位作者 SUN Xia WEI Qinsheng ZHANG Xuelei 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2016年第8期16-21,共6页
Harmful algal blooms(HABs) have been increasingly frequent in coastal waters around the world over the last several decades. Accelerated coastal eutrophication, resulting from the increased anthropogenic loadings of... Harmful algal blooms(HABs) have been increasingly frequent in coastal waters around the world over the last several decades. Accelerated coastal eutrophication, resulting from the increased anthropogenic loadings of nutrients, is commonly assumed to be the primary cause of this increase. However, although important,accelerated coastal eutrophication may not be the only explanation for the increasing blooms or toxic outbreaks in estuarine waters. Changes in riverine material fluxes other than nutrients, such as sediment load, may significantly affect biological activities and HAB incidence in estuarine and coastal waters. A case study off the Changjiang(Yangtze River) Estuary indicated that with the increasing riverine loadings of nutrients, the sediment load from the Changjiang River has been reduced by 70% over the past four decades. A comparison of long-term data revealed that the phytoplankton biomass maximum has expanded to a region of much lower salinity due to the drastic reduction in riverine sediment load and the subsequent improvement in light penetration in the Changjiang River plume. Furthermore, there was an apparent mirror-image relationship between the sediment load from the Changjiang River and the HAB incidence off the Changjiang Estuary over the past four decades, and the number of HAB incidents was significantly negatively correlated with the sediment load. Therefore, it is argued that the drastic decline in sediment load from the Changjiang River reduced turbidity in the Changjiang Estuary and thus contributed to the increased frequency of HABs in the buoyant discharge plumes. 展开更多
关键词 harmful algal blooms phytoplankton biomass sediment load chlorophyll Changjiang(Yangtze River) estuary
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Biodeposition, respiration, and excretion rates of an introduced clam Mercenaria mercenaria in ponds with implications for potential competition with the native clam Meretrix meretrix in Shuangtaizi estuary, China 被引量:1
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作者 张安国 袁秀堂 +4 位作者 侯文久 李晓东 赵凯 陈卫新 苏秀榕 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2016年第3期467-476,共10页
This study aimed to evaluate the potential impacts of an introduced clam Mercenaria mercenaria on estuarine ecosystem, and implications for the niche competition with a native clam Meretrix meretrix. The biodeposition... This study aimed to evaluate the potential impacts of an introduced clam Mercenaria mercenaria on estuarine ecosystem, and implications for the niche competition with a native clam Meretrix meretrix. The biodeposition, respiration, and excretion rates of 34. mercenaria were determined seasonally using a sediment trap and a closed respirator in field. The biodeposition rates ofM. mercenaria were 0.06-0.37 g/ (ind.·d), and the respiration rates were 0.31-14.66 mg/(ind.·d). The ammonia and phosphate excretion rates were 0.18-36.70 and 1.44-14.87 μg/(ind.·d), respectively. The hard clam M. mercenaria may discharge dry deposits up to 2.1 × 10^5 t, contribute 18.3 t ammonia and 9.0 t phosphate to culture ponds, and consume 7.9×10^3 t O2 from ponds annually. It suggested that the hard clam M. mercenaria might play an important role in pelagic-benthic coupling in pond ecosystem through biodeposition and excretion. A comparison of the key physiological parameters of the introduced clam M. mercenaria and the native clam Meretrix meretrix suggested that M. mercenaria had a niche similar to that of Meretrix meretrix in Shuangtaizi estuary and might have a potential competition with Meretrix meretrix for habitat and food ifM. mercenaria species escaped from the culture pond or artificially released in estuarine ecosystem. 展开更多
关键词 Mercenaria mercenaria Meretrix meretrix species introduction native clam physiological parameters potential competition Shuangtaizi estuary
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The impact of geomorphology of marsh creeks on fish assemblage in Changjiang River estuary 被引量:1
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作者 金斌松 许旺 +2 位作者 郭立 陈家宽 傅萃长 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2014年第2期469-479,共11页
Tidal marshes are an important habitat and nursery area for fi sh.In the past few decades,rapid economic development in the coastal areas of China has led to the interruption and destruction of an increasing number of... Tidal marshes are an important habitat and nursery area for fi sh.In the past few decades,rapid economic development in the coastal areas of China has led to the interruption and destruction of an increasing number of tidal marshes.The growing interest in tidal marsh restoration has increased the need to understand the relationship between geomorphological features and fi sh assemblages in the design of marsh restoration projects.We studied temporal variations in,and the effects of creek geomorphological features on,the estuarine tidal creek fi sh community.Using modifi ed channel nets,we sampled fi sh monthly from March 2007 to February 2008 from seven tidal creeks along an intertidal channel system in Chongming Dongtan National Nature Reserve.Fourteen creek geomorphological variables were measured or derived to characterize intertidal creek geomorphological features.The Gobiidae,with 10 species,was the most speciesrich family.The most abundant fi sh species were Liza affi nis,Chelon haematocheilus,and Lateolabrax maculatus.The fi sh community was dominated by juvenile marine transients,which comprised about 80% of the total catch.The highest abundance of fi sh occurred in June and July,and the highest biomass occurred in December.Canonical redundancy analyses demonstrated that depth,steepness,cross-sectional area,and volume signifi cantly affected the fi sh species assemblage.L.affi nis favored small creeks with high elevations.Synechogobius ommaturus,Acanthogobius luridus,and Carassius auratus preferred deep,steep creeks with a large cross-sectional area and volume.These fi ndings indicate that the geomorphological features of tidal creeks should be considered in the conservation and sustainable management of fi sh species and in the restoration of salt marshes. 展开更多
关键词 intertidal environment habitat selection GEOMORPHOLOGY RESTORATION salt marshes Changjiang (Yangtze) River estuary
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Composition of fish communities in an intertidal salt marsh creek in the Changjiang River estuary,China 被引量:1
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作者 全为民 倪勇 +1 位作者 施利燕 陈亚瞿 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2009年第4期806-815,共10页
Fish communities in a (third-order) intertidal creek in Dongtan marsh in the Changjiang (Yangtze) River estuary were investigated seasonally for one year. A total of 1 996 fish specimens (10 967.8 g) comprising ... Fish communities in a (third-order) intertidal creek in Dongtan marsh in the Changjiang (Yangtze) River estuary were investigated seasonally for one year. A total of 1 996 fish specimens (10 967.8 g) comprising 26 species and 15 families were collected. Abundances of fish communities in the intertidal salt marsh creek were primarily dominated by Boleophthalmus pectinirostris (19.8%), Collichthys lucidus (18.6%), Periophthalmus magnuspinnatus (18.2%), Liza haematocheilus (17.9%), and secondarily by Mugilogobius abel (8.5%), L. carinatus (7.2%), Odontamblyopus lacepedii (4.3%), and Acanthogobius ommaturus (3.9%); another 18 species were present only occasionally. Non-MDS ordination and SIMPER analysis indicated that there were two fish communities in the intertidal salt marsh creek. In spring, the communities were dominated by B. pectinirostris, P. magnuspinnatus, C. lucidus and M. abei; in summer, autumn, and winter by L. haematocheilus, L. carinatus, A. ommaturus and O. lacepedii. Some species showed strong habitat selection; L. carinatus and P magnuspinnatus were distributed mainly in the upper and middle creek, while B. pectinirostris, M. abei and O. lacepedii inhabited the middle and lower creek. The study indicated that the salt marshes of the Changjiang River estuary are an important nursery and feeding habitat for many fishes and should be protected. 展开更多
关键词 fish communities HABITAT NURSERY tidal creek salt marsh Changjiang (Yangtze) River estuary
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Stability of the Stratification of Water Bodies in the North Passage of the Yangtze River Estuary Based on the EFDC Model 被引量:1
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作者 WU De’an LI Xin WANG Yigang 《Journal of Ocean University of China》 CAS CSCD 2024年第1期23-32,共10页
To study the stratified stability of a water column in the North Passage of the Yangtze River Estuary,a numerical model of the hydrodynamics of this estuary is established using the EFDC model.On the basis of EFDC res... To study the stratified stability of a water column in the North Passage of the Yangtze River Estuary,a numerical model of the hydrodynamics of this estuary is established using the EFDC model.On the basis of EFDC results,this paper derives and pro-vides the discriminative index of water body stability caused by salinity and analyzes the along-range variation in water body strati-fication stability in the North Passage of the Yangtze River Estuary and the periodic variation at a key location(bend area)based on the simulation results of the numerical model.This work shows that the water body in the bend area varies between mixed and strati-fied types,and the vertical average flow velocity has a good negative correlation with the differential velocity between the surface and bottom layers of the water body.The model simulation results validate the formulae for the stratified stability discriminant during spring tides. 展开更多
关键词 Yangtze estuary North Passage EFDC model stratification stability salinity distribution
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Wnt1-Cre和Pax2-Cre标记的小鼠第一鳃弓颅颌面部神经嵴细胞异质性研究
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作者 徐珏 刘双 +3 位作者 符宏高 邵美瑛 陈美玲 黄镇 《华西口腔医学杂志》 CAS CSCD 北大核心 2024年第4期435-443,共9页
目的利用Wnt1-Cre和Pax2-Cre小鼠特异性标记颅颌面神经嵴细胞(CNCs)迁移到第一鳃弓时的分化异质性及机制。方法分别收取胚胎期(E)8.0~E9.25Wnt1-Cre;R26R^(mTmG)及Pax2-Cre;R26R^(mTmG)小鼠胚胎进行整体荧光观察,利用石蜡切片免疫荧光对... 目的利用Wnt1-Cre和Pax2-Cre小鼠特异性标记颅颌面神经嵴细胞(CNCs)迁移到第一鳃弓时的分化异质性及机制。方法分别收取胚胎期(E)8.0~E9.25Wnt1-Cre;R26R^(mTmG)及Pax2-Cre;R26R^(mTmG)小鼠胚胎进行整体荧光观察,利用石蜡切片免疫荧光对E15.5的Pax2-Cre;R26R^(Ai9)和Wnt1-Cre;R26R^(Ai9)小鼠所标记的CNCs在颅面部主要组织器官中的谱系分化情况进行比较分析,最后对E10.5的Wnt1-Cre;R26R^(mTmG)和Pax2-Cre;R26R^(mTmG)小鼠的第一鳃弓组织中CNCs进行单细胞测序分析,并对差异基因进行荧光定量聚合酶链反应(q-PCR)验证。结果Pax2-Cre和Wnt1-Cre小鼠特异性标记的CNCs均在E8.0自神经板开始迁移,但Pax2-Cre小鼠仅标记迁移到第一鳃弓的CNCs,而Wnt1-Cre同时标记了迁移到第一和第二鳃弓的CNCs;在分化谱系示踪方面,二者皆标记了CNCs分化形成的颅颌面部组织器官的间充质,但Wnt1-Cre在上腭和舌中标记CNCs更多;在第一鳃弓间充质中,Pax2-Cre所标记的CNCs特异性表达基因主要参与了成骨,而Wnt1-Cre所标记的CNCs特异性表达基因主要参与了肢体发育、细胞迁移和成骨,q-PCR结果也证实了两者高表达差异基因参与了以上功能。结论本研究结果提示Pax2-Cre小鼠可特异性用于第一鳃弓CNCs及其衍生组织成骨方面的研究。 展开更多
关键词 颅颌面部神经嵴细胞 第一鳃弓 Wnt1-cre Pax2-cre 单细胞测序
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Diel,seasonal,and annual variations of fish assemblages in intertidal creeks of the Changjiang River estuary
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作者 Jiayi FAN Jiasheng YANG +1 位作者 Yuanwen HE Xiaodong JIANG 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2023年第5期1849-1863,共15页
The use patterns of salt marsh habitats by fish assemblages were investigated in two welldeveloped intertidal creeks in the Changjiang(Yangtze)River estuary on spatial and temporal scales.Samples were collected using ... The use patterns of salt marsh habitats by fish assemblages were investigated in two welldeveloped intertidal creeks in the Changjiang(Yangtze)River estuary on spatial and temporal scales.Samples were collected using fyke nets at two sites during day and night in each season throughout four years.Notable changes in fish assemblages were detected over seasonal and interannual cycles,with many more marine species present in winter assemblages,whereas freshwater and estuarine species characterized the other seasonal samplings.The appearance of catadromous species in intertidal creeks mainly depended upon their specific physiological requirements,such as spawning migration.No significant diel shifts were observed in fish assemblages.This diel pattern may be a combined consequence of avian predation pressure and other drivers(i.e.,inherent living behaviors,food availability,and predation by piscivorous fishes).The spatial difference in fish community was insignificant even though the two intertidal creeks were different in size.Intertidal creeks with different size could possess similar ecological value for fish fauna.Redundancy analysis(RDA)showed that biological factors exerted greater effects on fish assemblages than physical variables.Temporal variations were strongly facilitated by food availability,but negatively impacted by avian predation.Salinity,water temperature,and river runoff had weak influence on temporal variations of fish communities.The lengthfrequency distribution showed that the fish collected in these two sites mainly consisted of young-of-year and juvenile fish,which confirmed that the creeks provided important nursery and forage roles in the process of fish ontogenetic development.Further,we proposed that salt marsh conservation management should be equally applied to all well-developed intertidal creeks regardless of creek size. 展开更多
关键词 FISH temporal variation intertidal creeks Changjiang(Yangtze)River estuary
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基于CRISPR/Cas9联合Cre-LoxP方法的Ddx3x肝脏条件性敲除小鼠模型的构建与鉴定
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作者 潘桢桢 徐玲 +5 位作者 张向颖 高耀 田原 范子豪 曹亚玲 任锋 《胃肠病学和肝病学杂志》 CAS 2024年第10期1275-1280,共6页
目的采用CRISPR/Cas9联合Cre-LoxP方法构建DEAD-box RNA解旋酶3X连锁(DEAD-box RNA helicase 3X-linked,Ddx3x)肝脏条件性敲除小鼠。方法设计特异的sgRNA序列,在Ddx3x基因外显子4~10两端插入LoxP序列,构建Ddx3x-flox小鼠。采用PCR方法... 目的采用CRISPR/Cas9联合Cre-LoxP方法构建DEAD-box RNA解旋酶3X连锁(DEAD-box RNA helicase 3X-linked,Ddx3x)肝脏条件性敲除小鼠。方法设计特异的sgRNA序列,在Ddx3x基因外显子4~10两端插入LoxP序列,构建Ddx3x-flox小鼠。采用PCR方法对F0代和F1代小鼠进行基因鉴定。将Ddx3x-flox F1代小鼠与特异性表达Alb-Cre的工具鼠交配,获得肝脏条件性敲除Ddx3x基因小鼠(Ddx3x^(Δhep))。采用PCR方法和Western blotting方法分别检测Ddx3x^(Δhep)基因小鼠目的基因与蛋白的表达。选取8周龄雄性野生型C57/6J小鼠、Ddx3x^(Δhep)小鼠、Ddx3x^(fl/fl)小鼠,测定血清ALT、AST水平评价肝脏肝损伤情况。结果对F1代小鼠基因PCR鉴定的结果表明,基因型符合Ddx3x^(fl/fl);F1代小鼠与Alb-Cre小鼠杂交,子代与Ddx3x^(fl/fl)小鼠杂交,即获得Ddx3x^(Δhep)小鼠;PCR与Western blotting结果显示,目的小鼠肝组织中Ddx3x基因和蛋白表达显著降低。此外,Ddx3x^(Δhep)小鼠无胚胎致死现象,出生后生理状况良好。正常饲养条件下,Ddx3x^(Δhep)小鼠与Ddx3x^(fl/fl)小鼠及野生型小鼠相比,血清ALT、AST指标及体质量差异均无统计学意义(P>0.05)。结论基于CRISPR/Cas9联合Cre-LoxP方法成功构建了Ddx3x^(Δhep)小鼠动物模型,为后续研究Ddx3x在肝脏中的生物学功能及作用机制奠定了良好的基础。 展开更多
关键词 CRISPR/Cas9技术 cre-LoxP系统 Ddx3x 肝脏条件性敲除小鼠
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采用Cre-loxP技术构建肝脏特异性Trappc11基因敲除小鼠模型
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作者 郑帆帆 李霞 +3 位作者 高枫 李梦瑶 杨彦玲 李优磊 《肝脏》 2024年第9期1137-1140,1153,共5页
目的采用Cre-loxP系统构建肝脏组织特异性Trappc11基因敲除小鼠(Trappc11^(flox/flox)-Alb-Cre^(+/-))模型以探究脂肪肝的发病机制。方法用带有loxP位点的Trappc11纯合子小鼠(Trappc11^(flox/flox))和带有Alb-Cre的转基因小鼠(Alb-Cre^(... 目的采用Cre-loxP系统构建肝脏组织特异性Trappc11基因敲除小鼠(Trappc11^(flox/flox)-Alb-Cre^(+/-))模型以探究脂肪肝的发病机制。方法用带有loxP位点的Trappc11纯合子小鼠(Trappc11^(flox/flox))和带有Alb-Cre的转基因小鼠(Alb-Cre^(+/-))进行杂交,对其子代进行基因型鉴定。筛选出Trappc11^(flox/+)-Alb-Cre^(+/-)小鼠,将其与Trappc11^(flox/flox)小鼠进一步杂交可获得Trappc11肝脏特异性敲除鼠。将3~4周龄的Trappc11^(flox/flox)-Alb-Cre^(+/-)、Trappc11^(flox/+)-Alb-^(Cre+/-)和Trappc11^(flox/flox)-Alb-^(Cre-/-)小鼠各3只,提取各组织脏器中的DNA,检测Trappc11的敲除效率和特异性。然后,在mRNA和蛋白质水平进一步验证。于第4周开始监测摄食量和体重等指标。结果我们成功繁育出肝脏特异性Trappc11基因敲除鼠,敲除心、脾、肺、肾、胰腺、肌肉、脂肪和脑组织中该基因表达未受影响,mRNA水平敲除效率达85%以上且TRAPPC11蛋白水平下降。该敲除鼠目前生长状态良好,饮食饮水正常,可用于后期繁殖与机制研究。结论已经构建成功的Trappc11^(flox/flox)-Alb-Cre^(+/-)鼠将为探索Trappc11在肝脏中的生理病理作用提供在体模型。 展开更多
关键词 转运蛋白颗粒复合物亚基11 cre-loxP重组酶系统 组织特异性敲除
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Cre/loxP条件性基因敲除小鼠在椎间盘退变研究中的应用
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作者 黄龙鳌 陈棣 江华 《中国脊柱脊髓杂志》 CAS CSCD 北大核心 2024年第9期990-997,共8页
椎间盘退变(intervertebral disc degeneration,IDD)是一个在生命早期就开始的过程,其与遗传、机械负荷、创伤和营养因素以及衰老密切相关,已证是腰痛的主要原因之一[1]。然而,IDD的分子生物学机制仍然不清楚。椎间盘的不同细胞类型表... 椎间盘退变(intervertebral disc degeneration,IDD)是一个在生命早期就开始的过程,其与遗传、机械负荷、创伤和营养因素以及衰老密切相关,已证是腰痛的主要原因之一[1]。然而,IDD的分子生物学机制仍然不清楚。椎间盘的不同细胞类型表达不同的基因,以调控椎间盘的发育和稳态。 展开更多
关键词 条件性基因敲除 椎间盘退变 生命早期 cre/loxP 分子生物学机制 营养因素 机械负荷 IDD
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Vav^(Cre)介导的荧光报告系统对小鼠小胶质细胞的标记效率
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作者 李冰冰 李佳倬 +2 位作者 朱盼盼 王一婷 谢华锋 《郑州大学学报(医学版)》 CAS 北大核心 2024年第4期450-454,共5页
目的:探讨Vav^(Cre)介导的荧光报告系统对小胶质细胞的标记效率。方法:将Vav^(Cre)/Rosa26REYFP fl/fl雄鼠与C57BL/6雌鼠杂交获得Vav^(Cre)/Rosa26REYFP^(fl/-)和Rosa26REYFP^(fl/-)子代。以发现雌鼠阴道栓的时间标记为E0.5,流式细胞术... 目的:探讨Vav^(Cre)介导的荧光报告系统对小胶质细胞的标记效率。方法:将Vav^(Cre)/Rosa26REYFP fl/fl雄鼠与C57BL/6雌鼠杂交获得Vav^(Cre)/Rosa26REYFP^(fl/-)和Rosa26REYFP^(fl/-)子代。以发现雌鼠阴道栓的时间标记为E0.5,流式细胞术检测EYFP在E7.5~E8.25、E10~E10.5、E12~E12.5、E14~E14.5小鼠胚胎和新生小鼠小胶质细胞中的表达。结果:Vav^(Cre)/Rosa26REYFP^(fl/-)组E10~E10.5、E12~E12.5、E14~E14.5胚胎和新生小鼠小胶质细胞中EYFP阳性率高于Rosa26REYFP^(fl/-)组(P<0.001),且均>95.000%。Vav^(Cre)/Rosa26REYFP^(fl/-)E7.5~E8.25小鼠胚胎中EYFP阳性率为(0.059±0.009)%。结论:以Vav^(Cre)介导的荧光报告系统可以稳定、高效地标记小鼠小胶质细胞,可以用于动态监测小胶质细胞的发育。 展开更多
关键词 小胶质细胞 Vav^(cre) Rosa26REYFP 小鼠
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基于Cre/Loxp系统的Csf1r-Cre^(ERT2)R26R^(EYFP)报告基因小鼠的构建及效率检测
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作者 朱向玲 吴旭铭 +5 位作者 王卉卉 周园园 王安琪 张慧茹 刘崇 涂佳杰 《安徽医科大学学报》 CAS 北大核心 2024年第7期1175-1180,共6页
目的构建报告基因小鼠,评价Csf1r-Cre^(ERT2)介导增强黄色荧光素蛋白EYFP标记组织CD45^(+)细胞CSF1R的效率。方法Csf1r-Cre^(ERT2)小鼠与R26R^(EYFP)小鼠繁育,他莫昔芬诱导、PCR筛选Csf1r-Cre^(ERT2)R26R^(EYFP)小鼠,流式细胞术和Wester... 目的构建报告基因小鼠,评价Csf1r-Cre^(ERT2)介导增强黄色荧光素蛋白EYFP标记组织CD45^(+)细胞CSF1R的效率。方法Csf1r-Cre^(ERT2)小鼠与R26R^(EYFP)小鼠繁育,他莫昔芬诱导、PCR筛选Csf1r-Cre^(ERT2)R26R^(EYFP)小鼠,流式细胞术和Western blot分析EYFP对不同组织以及不同组织CD45^(+)细胞中CSF1R的标记效率。结果获得Csf1r-Cre^(ERT2)R26R^(EYFP)报告基因小鼠。此外,Csf1r-Cre^(ERT2)小鼠介导EYFP可有效标记小鼠组织CSF1R以及不同部位中CD45^(+)细胞。与R26R^(EYFP)组比较,Csf1r-Cre^(ERT2)小鼠介导EYFP标记效率最高的是脑组织(P<0.001),最低的是胸腺组织(P<0.05),脾脏组织则差异无统计学意义。结论成年Csf1r-Cre^(ERT2)小鼠与R26R^(EYFP)小鼠是获得Csf1r-Cre^(ERT2)R26R^(EYFP)诱导型条件性荧光小鼠的有效途径。Csf1r-Cre^(ERT2)介导EYFP可对小鼠不同部位CSF1R以及CD45^(+)细胞中CSF1R进行有效示踪。 展开更多
关键词 Csf1r-cre^(ERT2) R26R^(EYFP) cre/LOXP系统 CD45 流式细胞术
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Inter-annual variations of dissolved oxygen and hypoxia off the northern Changjiang River(Yangtze River) Estuary in summer from 1997 to 2014
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作者 Anqi Liu Feng Zhou +6 位作者 Xiao Ma Qiang Zhao Guanghong Liao Yuntao Zhou Di Tian Xiaobo Ni Ruibin Ding 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2024年第6期119-130,共12页
Hypoxia off the Changjiang River Estuary has been the subject of much attention,yet systematic observations have been lacking,resulting in a lack of knowledge regarding its long-term change and drivers.By revisiting t... Hypoxia off the Changjiang River Estuary has been the subject of much attention,yet systematic observations have been lacking,resulting in a lack of knowledge regarding its long-term change and drivers.By revisiting the repeated surveys of dissolved oxygen(DO) and other relevant hydrographic parameters along the section from the Changjiang River Estuary to the Jeju-do in the summer from 1997 to 2014,rather different trends were revealed for the dual low-DO cores.The nearshore low-DO core,located close to the river mouth and relatively stable,shows that hypoxia has become more severe with the lowest DO descen ding at a rate of -0.07 mg/(L·a) and the thickness of low-DO zone rising at a rate of 0.43 m/a.The offshore core,centered around 40-m isobath but moving back and forth between 123.5°-125°E,shows large fluctuations in the minimum DO concentration,with the thickness of low-DO zone falling at a rate of -1.55 m/a.The probable factors affecting the minimum DO concentration in the two regions also vary.In the nearshore region,the decreasing minimum DO is driven by the increase in both stratification and primary productivity,with the enhanced extension of the Changjiang River Diluted Water(CDW) strengthening stratification.In the offshore region,the fluctuating trend of the minimum DO concentration indicates that both DO loss and DO supplement are distinct.The DO loss is primarily attributed to bottom apparent oxygen utilization caused by the organic matter decay and is also relevant to the advection of low-DO water from the nearshore region.The DO supplement is primarily due to weakened stratification.Our analysis also shows that the minimum DO concentration in the nearshore region was extremely low in 1998,2003,2007 and 2010,related to El Ni?o signal in these summers. 展开更多
关键词 dissolved oxygen low-DO Changjiang River estuary interannual variations DUAL-CORE
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Effects of phosphorus limitation on sinking velocities of phytoplankton during summer in the Changjiang River Estuary
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作者 Xinchi You Qiang Hao +6 位作者 Jie Zhu Wei Zhang Haiyan Jin Dewang Li Huanhong Ji Yu Ke Feng Zhou 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2024年第6期131-141,共11页
The sinking of phytoplankton is critical to organic matter transportation in the ocean and it is an essential process for the formation of coastal hypoxic zones.This study was based on a field investigation conducted ... The sinking of phytoplankton is critical to organic matter transportation in the ocean and it is an essential process for the formation of coastal hypoxic zones.This study was based on a field investigation conducted during the summer of 2022 in the Changjiang River(Yangtze River) Estuary(CJE) and its adjacent waters.The settling column method was employed to measure the sinking velocity(SV) of different size fractions of phytoplankton at the surface of the sea and to analyze their environmental control mechanisms.The findings reveal significant spatial variation in phytoplankton SV(-0.55-2.41 m/d) within the CJE.High-speed sinking was predominantly observed in phosphate-depleted regions beyond the CJE front.At the same time,an upward trend was more commonly observed in the phosphate-rich regions near the CJE mouth.The SV ranges for different sizefractionated phytoplankton,including micro-(>20 μm),nano-(2-20 μm),and picophytoplankton(0.7-2 μm),were-0.50-4.74 m/d,-1.04-1.59 m/d,and-1.24-1.65 m/d,respectively.Correlation analysis revealed a significant negative correlation between SV and dissolved inorganic phosphorus(DIP),implying that the influence of DIP contributes to SV.The variations in phytoplankton alkaline phosphatase activity suggested a significant increase in SV across all size fractions in the event of phosphorus limitation.Phytoplankton communities with limited photo synthetic capacity(maximum photochemical efficience,Fv/Fm <0.3) were found to have higher SV than that of communities with strong capacity,suggesting a link between sinking and alterations in physiological conditions due to phosphate depletion.The findings from the in situ phosphate enrichment experiments confirmed a marked decrease in SV following phosphate supplementation.These findings suggest that phosphorus limitation is the primary driver of elevated SV in the CJE.This study enhances the comprehension of the potential mechanisms underlying hypoxic zone formation in the CJE,providing novel insights into how nearshore eutrophication influences organic carbon migration. 展开更多
关键词 PHYTOPLANKTON sinking velocity Changjiang River estuary phosphorus limitation alkaline phosphatase
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Tetrabromobisphenol A and hexabromocyclododecane in sediments from the Zhujiang(Pearl) River Estuary and South China Sea
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作者 Chuyue Long Weiyan Yang +6 位作者 Jiaxun Lu Yuanyue Cheng Ning Qiu Sen Du Li Zhang Shejun Chen Yuxin Sun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2024年第6期32-37,共6页
Marine sediments collected from the Zhujiang(Pearl) River Estuary(ZRE) and South China Sea(SCS) were utilized to study the occurrence and spatial distribution of tetrabromobisphenol A(TBBPA) and hexabromocyclododecane... Marine sediments collected from the Zhujiang(Pearl) River Estuary(ZRE) and South China Sea(SCS) were utilized to study the occurrence and spatial distribution of tetrabromobisphenol A(TBBPA) and hexabromocyclododecane(HBCDD).The levels of TBBPA and HBCDD in sediments ranged from not detected(nd) to 6.14 ng/g dry weight(dw) and nd to 0.42 ng/g dw.TBBPA concentrations in marine sediments were substantially higher than HBCDD.The concentrations of TBBPA and HBCDD in the ZRE sediments were significantly greater than those in the SCS.α-HBCDD(48.7%) and γ-HBCDD(46.2%) were the two main diastereoisomers of HBCDD in sediments from the ZRE,with minor contribution of β-HBCDD(5.1%).HBCDD were only found in one sample from the northern SCS.The enantiomeric fraction of α-HBCDD in sediments from the ZRE was obviously greater than 0.5,indicating an accumulation of(+)-α-HBCDD.The enantiomers of HBCDD were not measured in sediments from the SCS.This work highlighted the environmental behaviors of TBBPA and HBCDD in marine sediments. 展开更多
关键词 tetrabromobisphenol A HEXABROMOCYCLODODECANE marine sediments Zhujiang River estuary South China Sea
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Coastal hypoxia response to the coupling of catastrophic flood,extreme marine heatwave and typhoon:a case study off the Changjiang River Estuary in summer 2020
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作者 Xiao Ma Qicheng Meng +8 位作者 Dewang Li Yuanli Zhu Xiaobo Ni Dingyong Zeng Di Tian Ting Huang Zhihao Jiang Haiyan Jin Feng Zhou 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2024年第6期107-118,共12页
Massive bodies of low-oxygen bottom waters are found in coastal areas worldwide,which are detrimental to coastal ecosystems.In summer 2020,the response of coastal hypoxia to extreme weather events,including a catastro... Massive bodies of low-oxygen bottom waters are found in coastal areas worldwide,which are detrimental to coastal ecosystems.In summer 2020,the response of coastal hypoxia to extreme weather events,including a catastrophic flooding,an extreme marine heatwave,and Typhoon Bavi,is investigated based on multiple satellite,four cruises,and mooring observations.The extensive fan-shaped hypoxia zone presents significant northward extension during July-September 2020,and is estimated as large as 13 000 km^(2) with rather low oxygen minimum(0.42 mg/L) during its peak in 28-30 August.This severe hypoxia is attributed to the persistent strong stratification,which is indicated by flood-induced larger amount of riverine freshwater input and subsequent marine heatwave off the Changjiang River Estuary.Moreover,the Typhoon Bavi has limited effect on the marine heatwave and coastal hypoxia in summer 2020. 展开更多
关键词 coastal hypoxia Changjiang River estuary extreme weather events seasonal evolution
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Biogeochemistry of sedimentary organic matter in the Yongjiang River estuary in the southern part of Hangzhou Bay,China,since the Late Pleistocene
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作者 Dongqin HUANG Xiaolong LI +6 位作者 Zilong LI Pei Sun LOH Jianxiong HU Jianfang CHEN Yuan-Pin CHANG Chin-Wen YANG Qin GAO 《Journal of Oceanology and Limnology》 SCIE CAS CSCD 2024年第3期772-786,共15页
A sediment core(YJK19-02)collected from the southern outlet of Hangzhou Bay near the Yongjiang River estuary in East China was analyzed for grain size,lignin,bulk elemental composition,stable carbon isotope,and rare e... A sediment core(YJK19-02)collected from the southern outlet of Hangzhou Bay near the Yongjiang River estuary in East China was analyzed for grain size,lignin,bulk elemental composition,stable carbon isotope,and rare earth elements(REEs)to determine the sources and diagenesis of sedimentary organic matter(OM)of the estuary and adjacent areas since the Late Pleistocene.δ^(13)C values(-24.80‰–-23.60‰),total organic carbon/total nitrogen(TOC/TN)molar ratios(8.00–12.14),and light rare earth element/heavy rare earth element ratios(LREE/HREE=8.34–8.91)revealed the predominance of terrestrial sources of OM,mainly from the Changjiang(Yangtze)River.The lignin parameters of syringyl/vanillyl(S/V=0.20–0.73)and cinnamyl/vanillyl(C/V=0.03–0.19)ratios indicate the predominance of nonwoody angiosperms,and the vanillic acid/vanillin ratios[(Ad/Al)_(V)=0.32–1.57]indicate medium to high degrees of lignin degradation.An increasing trend ofΛ(total lignin in mg/100-mg OC)values from ca.14500 a BP to ca.11000 a BP reflected the increase in temperature during the Late Pleistocene.However,a time lag effect of temperature on vegetation abundance was also revealed.The relatively higher and stableΛvalues correspond to the higher temperature during the mid-Holocene from ca.8500 a BP to ca.4500 a BP.Λvalues decreased from ca.4000 a BP to the present,corresponding to historical temperature fluctuations during this time.Our results show that the vegetation abundance in the Yongjiang River Basin since the Late Pleistocene was related to the temperature fluctuation duo to climate change. 展开更多
关键词 Yongjiang River estuary sedimentary organic matter LIGNIN environmental change Late Pleistocene
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