Lake Vrana on the island of Cres is one of the largest fresh water features on Mediterranean islands. The maximum depth of the lake is 72 m and it stores 220 million m3 of fresh water. The paper provides an overview o...Lake Vrana on the island of Cres is one of the largest fresh water features on Mediterranean islands. The maximum depth of the lake is 72 m and it stores 220 million m3 of fresh water. The paper provides an overview of lake and groundwater temperature measurements to date and recent activities within the EU project ＂CC-WaterS （Climate Change and Impacts on Water Supply）＂. Groundwater temperatures in the lake surrounding are almost constant throughout the year, in the range from 14.6 ℃ to 13.1 ℃, while thermodynamic cycle of the lake conforms to the characteristics of a monomictic, medium depth lake in the moderate climate belt. Present and future climate simulations using three limited area models were analyzed （Aladin, Promes and RegCM3）, they pointed out further air temperature increase in range of 0.27 ℃/10 yrs to 0.32 ℃/10 yrs. The significant changes of precipitation rates were not indicated. Considering increasing water consumption from the lake, already asserted negative trends, indicated climate changes and possible effects on the lake recharge, it is necessary to establish continual monitoring of parameters that describe lake system behaviour and periodically analyze lake conditions, especially with respect to the extraction for the public water supply.展开更多
The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Re...The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Research,Vol.25,No.4,2017,pp.595–603.展开更多
Precise chromosome engineering has traditionally relied on the Cre-Lox recombination system-an approach in which the enzyme Cre functions like molecular scissors,cutting and rejoining DNA at specific“Lox”sites to ad...Precise chromosome engineering has traditionally relied on the Cre-Lox recombination system-an approach in which the enzyme Cre functions like molecular scissors,cutting and rejoining DNA at specific“Lox”sites to add,remove,or flip genomic DNA segments inside living cells.展开更多
Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genet...Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genetic tools to manipulate neurons in rats could benefit neurological research.Methods:Using the CRISPR/Cas9 system,we inserted a Cre cassette into endogenous Thy1 and NeuN loci.Thy1-Cre rats featured a downstream P2A-linked insertion,while NeuN-Cre was inserted at the transcriptional start site.The Cre activity was assessed by crossing with a Cre reporter(Rosa26 imCherry)rat and through analyzing mCherry expression patterns.The specificity of cell type was further confirmed by immunofluorescence with NeuN antibody.Phenotypic consequences were assessed by crossing with ND1^(LSL) rats to deplete ND1,followed by monitoring weight/survival and conducting motor function tests.Results:We generated two neuron-specific rats(Thy1-Cre and NeuN-Cre),which exhibited high neuron-specific Cre expression in brain and spinal cord with minor leakage in other tissues.Thy1-Cre showed minor leakage in spleen,lung and kidney while NeuN-Cre showed minor leakage in spleen and kidney.ND1^(Thy1-Cre) and ND1^(NeuN-Cre) rats both showed decreased body weights and survival times.The ND1^(NeuN-Cre) rats died within two weeks,while ND1^(Thy1-Cre) rats lived longer with impaired motor function.Conclusions:We successfully generated two neuron-specific NeuN-Cre and Thy1-Cre rats,and systemically analyzed their expression pattern.展开更多
Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed ra...Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.展开更多
文摘Lake Vrana on the island of Cres is one of the largest fresh water features on Mediterranean islands. The maximum depth of the lake is 72 m and it stores 220 million m3 of fresh water. The paper provides an overview of lake and groundwater temperature measurements to date and recent activities within the EU project ＂CC-WaterS （Climate Change and Impacts on Water Supply）＂. Groundwater temperatures in the lake surrounding are almost constant throughout the year, in the range from 14.6 ℃ to 13.1 ℃, while thermodynamic cycle of the lake conforms to the characteristics of a monomictic, medium depth lake in the moderate climate belt. Present and future climate simulations using three limited area models were analyzed （Aladin, Promes and RegCM3）, they pointed out further air temperature increase in range of 0.27 ℃/10 yrs to 0.32 ℃/10 yrs. The significant changes of precipitation rates were not indicated. Considering increasing water consumption from the lake, already asserted negative trends, indicated climate changes and possible effects on the lake recharge, it is necessary to establish continual monitoring of parameters that describe lake system behaviour and periodically analyze lake conditions, especially with respect to the extraction for the public water supply.
文摘The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Research,Vol.25,No.4,2017,pp.595–603.
文摘Precise chromosome engineering has traditionally relied on the Cre-Lox recombination system-an approach in which the enzyme Cre functions like molecular scissors,cutting and rejoining DNA at specific“Lox”sites to add,remove,or flip genomic DNA segments inside living cells.
基金Research Project of China Baoyuan Investment Co.,Ltd,Grant/Award Number:Program CBYI202102Haihe Laboratory of Cell Ecosystem Innovation Fund,Grant/Award Number:HH24KYZX0007+4 种基金CAMS Innovation Fund for Medical Sciences,Grant/Award Number:2021-I2M-1-024,2021-I2M-1-034 and 2023-I2M-2-001Fundamental Research Funds for the Central Universities,Grant/Award Number:3332022040 and 3332023164Open Research Project in State Key Laboratory of Vascular Homeostasis and Remodeling,Peking University,Grant/Award Number:202411State Key Laboratory Special Fund,Grant/Award Number:2060204the Non-profit Central Research Institute Fund of the Chinese Academy of Medical Sciences,Grant/Award Number:2023-PT180-01.
文摘Background:The Cre/loxP system is most popular in mice,but its application in rats has largely lagged far behind.The rat is vital laboratory animal,especially in toxicological and neurological studies.Generating genetic tools to manipulate neurons in rats could benefit neurological research.Methods:Using the CRISPR/Cas9 system,we inserted a Cre cassette into endogenous Thy1 and NeuN loci.Thy1-Cre rats featured a downstream P2A-linked insertion,while NeuN-Cre was inserted at the transcriptional start site.The Cre activity was assessed by crossing with a Cre reporter(Rosa26 imCherry)rat and through analyzing mCherry expression patterns.The specificity of cell type was further confirmed by immunofluorescence with NeuN antibody.Phenotypic consequences were assessed by crossing with ND1^(LSL) rats to deplete ND1,followed by monitoring weight/survival and conducting motor function tests.Results:We generated two neuron-specific rats(Thy1-Cre and NeuN-Cre),which exhibited high neuron-specific Cre expression in brain and spinal cord with minor leakage in other tissues.Thy1-Cre showed minor leakage in spleen,lung and kidney while NeuN-Cre showed minor leakage in spleen and kidney.ND1^(Thy1-Cre) and ND1^(NeuN-Cre) rats both showed decreased body weights and survival times.The ND1^(NeuN-Cre) rats died within two weeks,while ND1^(Thy1-Cre) rats lived longer with impaired motor function.Conclusions:We successfully generated two neuron-specific NeuN-Cre and Thy1-Cre rats,and systemically analyzed their expression pattern.
基金CAMS Innovation Fund for Medical Sciences(CIFMS),Grant/Award Number:2021-I2M-1-035National Natural Science Foundation of China,Grant/Award Number:31970508。
文摘Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.
