The halopyrimidine 5-bromo-2′-deoxyuridine(BrdU)is an exogenous marker of DNA synthesis.Since the introduction of monoclonal antibodies against BrdU,an increasing number of methodologies have been used for the immuno...The halopyrimidine 5-bromo-2′-deoxyuridine(BrdU)is an exogenous marker of DNA synthesis.Since the introduction of monoclonal antibodies against BrdU,an increasing number of methodologies have been used for the immunodetection of this synthesized bromine-tagged base analogue into replicating DNA.BrdU labeling is widely used for identifying neuron precursors and following their fate during the embryonic,perinatal,and adult neurogenesis in a variety of vertebrate species including birds,reptiles,and mammals.Due to BrdU toxicity,its incorporation into replicating DNA presents adverse consequences on the generation,survival,and settled patterns of cells.This may lead to false results and misinterpretation in the identification of proliferative neuroblasts.In this review,I will indicate the detrimental effects of this nucleoside during the development of the central nervous system,as well as the reliability of BrdU labeling to detect proliferating neuroblasts.Moreover,it will show factors influencing BrdU immunodetection and the contribution of this nucleoside to the study of prenatal,perinatal,and adult neurogenesis.Human adult neurogenesis will also be discussed.It is my hope that this review serves as a reference for those researchers who focused on detecting cells that are in the synthetic phase of the cell cycle.展开更多
The health effects of ambient PM 2.5 and its potential mechanisms have generated considerable interest.In vitro cell studies and ex vivo animal experiments may not accurately determine the characteristics of PM 2.5 pa...The health effects of ambient PM 2.5 and its potential mechanisms have generated considerable interest.In vitro cell studies and ex vivo animal experiments may not accurately determine the characteristics of PM 2.5 particles.To better understand their detailed mechanisms,we performed an in vivo study using single photon emission tomography(SPECT)imaging.To mimic the PM 2.5 particles,SiO2 nanoparticles modified by ethylene carbonate or polyvinyl pyrrolidone were labeled with 131I.After administration via inhalation,in vivo SPECT imaging of the radiolabeled particles in sprague dawley rats was performed.It was found that radioactivity accumulated in the lungs and trachea 6 and 24 h after administration.In addition,significant radioactivity was observed in the abdomen,including the liver and kidneys.The results were also confirmed by ex vivo autoradiography.This study revealed that in vivo SPECT imaging could be an effective method for investigating the properties of PM 2.5 particles.展开更多
Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by...Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase展开更多
BACKGROUND Type 2 diabetes mellitus(T2DM) has been strongly associated with an increased risk of developing cognitive dysfunction and dementia.The mechanisms of diabetes-associated cognitive dysfunction(DACD) have not...BACKGROUND Type 2 diabetes mellitus(T2DM) has been strongly associated with an increased risk of developing cognitive dysfunction and dementia.The mechanisms of diabetes-associated cognitive dysfunction(DACD) have not been fully elucidated to date.Some studies proved lower cerebral blood flow(CBF) in the hippocampus was associated with poor executive function and memory in T2DM.Increasing evidence showed that diabetes leads to abnormal vascular endothelial growth factor(VEGF) expression and CBF changes in humans and animal models.In this study,we hypothesized that DACD was correlated with CBF alteration as measured by three-dimensional(3D) arterial spin labeling(3D-ASL) and VEGF expression in the hippocampus.AIM To assess the correlation between CBF(measured by 3D-ASL and VEGF expression) and DACD in a rat model of T2DM.METHODS Forty Sprague-Dawley male rats were divided into control and T2DM groups.The T2DM group was established by feeding rats a high-fat diet and glucose to induce impaired glucose tolerance and then injecting them with streptozotocin to induce T2DM.Cognitive function was assessed using the Morris water maze experiment.The CBF changes were measured by 3D-ASL magnetic resonance imaging.VEGF expression was determined using immunofluorescence.RESULTS The escape latency time significantly reduced 15 wk after streptozotocin injection in the T2DM group.The total distance traveled was longer in the T2DM group;also,the platform was crossed fewer times.The percentage of distance in the target zone significantly decreased.CBF decreased in the bilateral hippocampus in the T2DM group.No difference was found between the right CBF value and the left CBF value in the T2DM group.The VEGF expression level in the hippocampus was lower in the T2DM group and correlated with the CBF value.The escape latency negatively correlated with the CBF value.The number of rats crossing the platform positively correlated with the CBF value.CONCLUSION Low CBF in the hippocampus and decreased VEGF expression might be crucial in DACD.CBF measured by 3D-ASL might serve as a noninvasive imaging biomarker for cognitive impairment associated with T2DM.展开更多
We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with E...We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with Eu^(3+) when conjugated to protein.The labeled proteins were characterized by absorbance and fluorescence measurements and the effect of labeling on the biological activity of sone proteins was also studied.It is shown that the new label is suitable for applications in time-resolved fluoroimmunoassay.展开更多
Objective： The combination of both nuclear and fluorescent reporters provides unique opportunities for noninvasive nuclear imaging with subsequent fluorescence image-guided resection and pathology. Our objective was ...Objective： The combination of both nuclear and fluorescent reporters provides unique opportunities for noninvasive nuclear imaging with subsequent fluorescence image-guided resection and pathology. Our objective was to synthesize and optimize a dual-labeled trastuzumab-based imaging agent that can be used to validate an optical imaging agent with potential use in identifying tumor metastases in human epidermal growth factor receptor 2 （HER2） positive breast cancer patients. Methods： [111In]-DTPA-trastuzumab-IRDye 800 was synthesized by a three-step procedure. Purity, stability, immunoreactivity, internalization and biodistribution were explored in HER2＋ SKBR-3 cells. Biodistribution of [111In]-DTPA-trastuzumab-IRDye 800 was performed in a SKBR-3 xenograft model. Results： [111In]-DTPA-trastuzumab-IRDye 800 demonstrated high purity by both chemical and fluorometric determinations. Both flow cytometry and the Lindmo assay demonstrated a high binding affinity of [111In]-DTPA-trastuzumab-IRDye 800 to HER2-overexpressing cells. The dual-labeled conjugate was stable in PBS, but not in serum after 24 h at 37 ℃. Larger molecules （〉150 kD） were seen after a 24 h-incubation in human serum. Biodistribution studies revealed tumor-specific accumulation of [111In]-DTPA- trastuzumab-IRDye 800 in SKBR-3 tumors, and tumor uptakes at 24 and 48 h were （12.42±1.72）% and （9.96±1.05） %, respectively, following intravenous administration. The tumor-to-muscle ratio was 9.13±1.68 at 24 h, and increased to 12.79±2.13 at 48 h. Liver and kidney showed marked uptake of the dual-labeled imaging agent. Conclusions： [111In]-DTPA-trastuzumab-IRDye 800 is an effective diagnostic biomarker that can be used to validate dual-labeled, molecularly targeted imaging agents and can allow these agents to be translated into clinical practice for identifying HER2＋ lesions.展开更多
11C-labeled C1-C10 partial structure of kulokekahilide-2 (1) was successfully synthesized based on Pd0-mediated rapid C-[11C]methylation using [11C]methyl iodide and pinacol alkenylboronate. The preparation of organob...11C-labeled C1-C10 partial structure of kulokekahilide-2 (1) was successfully synthesized based on Pd0-mediated rapid C-[11C]methylation using [11C]methyl iodide and pinacol alkenylboronate. The preparation of organoboron intermediate via olefin cross-metathesis is also a crucial procedure for the synthesis of 11C-labeling C1-C10 dihy-droxy acid moiety of 1.展开更多
A k-L(2,1)-labeling for a graph G is a function such that whenever and whenever u and v are at distance two apart. The λ-number for G, denoted by λ(G), is the minimum k over all k-L(2,1)-labelings of G. In this pape...A k-L(2,1)-labeling for a graph G is a function such that whenever and whenever u and v are at distance two apart. The λ-number for G, denoted by λ(G), is the minimum k over all k-L(2,1)-labelings of G. In this paper, we show that for or 11, which confirms Conjecture 6.1 stated in [X. Li, V. Mak-Hau, S. Zhou, The L(2,1)-labelling problem for cubic Cayley graphs on dihedral groups, J. Comb. Optim. (2013) 25: 716-736] in the case when or 11. Moreover, we show that? if 1) either (mod 6), m is odd, r = 3, or 2) (mod 3), m is even (mod 2), r = 0.展开更多
Objective： To investigate the effect of fluorescent dye labeling on the targeting capabilities of 111 In- （DTPA）.-trastuzumab-（IRDye 800）m. Methods： Trastuzumab-based conjugates were synthesized and conjugated w...Objective： To investigate the effect of fluorescent dye labeling on the targeting capabilities of 111 In- （DTPA）.-trastuzumab-（IRDye 800）m. Methods： Trastuzumab-based conjugates were synthesized and conjugated with diethylenetriaminepentaacetic acid （DTPA） at molar ratios of I, 2, 3 and 5 and with a fluorescent dye （IRDye 800CW） at molar ratios of 1, 3 and 5. Immunoreactivity and internalization were assessed on SKBR-3 cells, overexpressing human epidermal growth factor receptor 2. The stability in human serum and phosphate-buffered saline （PBS） was evaluated. The biodistribution of dual-labeled conjugates was compared with that of 111In-（DTPA）2-trastuzumab in a SKBR-3 xenograft model to evaluate the effect of dye-to-protein ratio. Results： All trastuzumab-based conjugates exhibited a high level of chemical and optical purity. Flow cytometry results showed that increasing dye-to-protein ratios were associated with decreased immunoreactivity. Stability studies revealed that the conjugate was stable in PBS, while in human serum, increased degradation and protein precipitation were observed with increasing dye-to-protein ratios. At 4 h, the percentages of internalization of dual-labeled conjugates normalized by dye-to-protein ratio （m） were 24.88%±2.10%, 19.99%±0.59%, and 17.47%±1.26% for ＂m＂ equal to 1, 3, and 5, respectively. A biodistribution study revealed a progressive decrease in tumor uptake with an increase in the dye-to-protein ratios. The liver, spleen and kidney showed a marked uptake with increased dye-to-protein ratios, particularly in the latter. Conclusions： With non-specific-site conjugation of the fluorescent dye with a protein based on imaging agent, the increase in dye-to-protein ratios negatively impacted the immunoreactivity and stability, indicating a reduced tumor uptake.展开更多
the widely discussed study of word class categorization has a long history of more than 2000 years,which is known as the study of the“God Particles”in language.As a typical analytic language,Modern Chinese,due to it...the widely discussed study of word class categorization has a long history of more than 2000 years,which is known as the study of the“God Particles”in language.As a typical analytic language,Modern Chinese,due to its lack of morphological changes,is challenged by a thorny problem of word classes especially when it comes to the criteria for word class identification and the treat⁃ment of multiple class membership.As such,all the controversies eventually give rise to some contradiction and confusion in word class labeling in Modern Chinese and Chinese-English dictionaries.As an important grammatical means in Chinese and the focus of lexicology and rhetorics,total reduplication lexemes serve as an essential part of Chinese-English dictionaries with complex and diverse word classes.Guided by the Two-level Word Class Categorization Theory,this thesis focuses on the word class labeling of total reduplication lexemes in New Century Chinese-English Dictionary(2nd edition)backed by large-scale balanced Modern Chi⁃nese corpora.With an innovative theoretical perspective,this study not only contributes to the word class labeling of total reduplica⁃tion lexemes and even sheds light on the compilation of Chinese-English dictionaries,but also drives the study of Modern Chinese word classes in the long term.展开更多
文摘The halopyrimidine 5-bromo-2′-deoxyuridine(BrdU)is an exogenous marker of DNA synthesis.Since the introduction of monoclonal antibodies against BrdU,an increasing number of methodologies have been used for the immunodetection of this synthesized bromine-tagged base analogue into replicating DNA.BrdU labeling is widely used for identifying neuron precursors and following their fate during the embryonic,perinatal,and adult neurogenesis in a variety of vertebrate species including birds,reptiles,and mammals.Due to BrdU toxicity,its incorporation into replicating DNA presents adverse consequences on the generation,survival,and settled patterns of cells.This may lead to false results and misinterpretation in the identification of proliferative neuroblasts.In this review,I will indicate the detrimental effects of this nucleoside during the development of the central nervous system,as well as the reliability of BrdU labeling to detect proliferating neuroblasts.Moreover,it will show factors influencing BrdU immunodetection and the contribution of this nucleoside to the study of prenatal,perinatal,and adult neurogenesis.Human adult neurogenesis will also be discussed.It is my hope that this review serves as a reference for those researchers who focused on detecting cells that are in the synthetic phase of the cell cycle.
基金supported by the National Natural Science Foundation of China(Nos.31671035,51803082)National Significant New Drugs Creation Program(No.2017ZX09304021)+1 种基金Jiangsu Province Foundation(Nos.BK20170204,BK20161137)Jiangsu Provincial Medical Innovation Team(Nos.CXTDA2017024,LGY2017088,QNRC2016628)。
文摘The health effects of ambient PM 2.5 and its potential mechanisms have generated considerable interest.In vitro cell studies and ex vivo animal experiments may not accurately determine the characteristics of PM 2.5 particles.To better understand their detailed mechanisms,we performed an in vivo study using single photon emission tomography(SPECT)imaging.To mimic the PM 2.5 particles,SiO2 nanoparticles modified by ethylene carbonate or polyvinyl pyrrolidone were labeled with 131I.After administration via inhalation,in vivo SPECT imaging of the radiolabeled particles in sprague dawley rats was performed.It was found that radioactivity accumulated in the lungs and trachea 6 and 24 h after administration.In addition,significant radioactivity was observed in the abdomen,including the liver and kidneys.The results were also confirmed by ex vivo autoradiography.This study revealed that in vivo SPECT imaging could be an effective method for investigating the properties of PM 2.5 particles.
文摘Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase
基金National Natural Science Foundation of China(No.10671074 and No.60673048)Natural Science Foundation of Education Ministry of Anhui Province(No.KJ2007B124 and No.2006KJ256B)
基金Supported by The Endocrine Clinical Medical Center of Yunnan ProvinceNo.ZX20190202+2 种基金the Fund of the Diabetic Innovation Team in Yunnan Province,No.2019HC002the Special Joint Fund from Yunnan Provincial Department of Science and Technology and Kunming Medical University,Kunming,Yunnan,China,No.2018FE001(-267)the SKY Image Research Fund,China,No. Z-2014-07-2003-12。
文摘BACKGROUND Type 2 diabetes mellitus(T2DM) has been strongly associated with an increased risk of developing cognitive dysfunction and dementia.The mechanisms of diabetes-associated cognitive dysfunction(DACD) have not been fully elucidated to date.Some studies proved lower cerebral blood flow(CBF) in the hippocampus was associated with poor executive function and memory in T2DM.Increasing evidence showed that diabetes leads to abnormal vascular endothelial growth factor(VEGF) expression and CBF changes in humans and animal models.In this study,we hypothesized that DACD was correlated with CBF alteration as measured by three-dimensional(3D) arterial spin labeling(3D-ASL) and VEGF expression in the hippocampus.AIM To assess the correlation between CBF(measured by 3D-ASL and VEGF expression) and DACD in a rat model of T2DM.METHODS Forty Sprague-Dawley male rats were divided into control and T2DM groups.The T2DM group was established by feeding rats a high-fat diet and glucose to induce impaired glucose tolerance and then injecting them with streptozotocin to induce T2DM.Cognitive function was assessed using the Morris water maze experiment.The CBF changes were measured by 3D-ASL magnetic resonance imaging.VEGF expression was determined using immunofluorescence.RESULTS The escape latency time significantly reduced 15 wk after streptozotocin injection in the T2DM group.The total distance traveled was longer in the T2DM group;also,the platform was crossed fewer times.The percentage of distance in the target zone significantly decreased.CBF decreased in the bilateral hippocampus in the T2DM group.No difference was found between the right CBF value and the left CBF value in the T2DM group.The VEGF expression level in the hippocampus was lower in the T2DM group and correlated with the CBF value.The escape latency negatively correlated with the CBF value.The number of rats crossing the platform positively correlated with the CBF value.CONCLUSION Low CBF in the hippocampus and decreased VEGF expression might be crucial in DACD.CBF measured by 3D-ASL might serve as a noninvasive imaging biomarker for cognitive impairment associated with T2DM.
基金Supported by National Natural Science Foundation of China.
文摘We describe for the first time the synthesis and the optimal conditions for protein labeling with a new fluorescent probe,5-chlorosulfoyl-2-thenoyltrifluoroacetone(CTTA),whicb forms a highly fluorescent conplex with Eu^(3+) when conjugated to protein.The labeled proteins were characterized by absorbance and fluorescence measurements and the effect of labeling on the biological activity of sone proteins was also studied.It is shown that the new label is suitable for applications in time-resolved fluoroimmunoassay.
基金supported by Beijing Natural Science Foundation (No. 7132037)NIH R01
文摘Objective： The combination of both nuclear and fluorescent reporters provides unique opportunities for noninvasive nuclear imaging with subsequent fluorescence image-guided resection and pathology. Our objective was to synthesize and optimize a dual-labeled trastuzumab-based imaging agent that can be used to validate an optical imaging agent with potential use in identifying tumor metastases in human epidermal growth factor receptor 2 （HER2） positive breast cancer patients. Methods： [111In]-DTPA-trastuzumab-IRDye 800 was synthesized by a three-step procedure. Purity, stability, immunoreactivity, internalization and biodistribution were explored in HER2＋ SKBR-3 cells. Biodistribution of [111In]-DTPA-trastuzumab-IRDye 800 was performed in a SKBR-3 xenograft model. Results： [111In]-DTPA-trastuzumab-IRDye 800 demonstrated high purity by both chemical and fluorometric determinations. Both flow cytometry and the Lindmo assay demonstrated a high binding affinity of [111In]-DTPA-trastuzumab-IRDye 800 to HER2-overexpressing cells. The dual-labeled conjugate was stable in PBS, but not in serum after 24 h at 37 ℃. Larger molecules （〉150 kD） were seen after a 24 h-incubation in human serum. Biodistribution studies revealed tumor-specific accumulation of [111In]-DTPA- trastuzumab-IRDye 800 in SKBR-3 tumors, and tumor uptakes at 24 and 48 h were （12.42±1.72）% and （9.96±1.05） %, respectively, following intravenous administration. The tumor-to-muscle ratio was 9.13±1.68 at 24 h, and increased to 12.79±2.13 at 48 h. Liver and kidney showed marked uptake of the dual-labeled imaging agent. Conclusions： [111In]-DTPA-trastuzumab-IRDye 800 is an effective diagnostic biomarker that can be used to validate dual-labeled, molecularly targeted imaging agents and can allow these agents to be translated into clinical practice for identifying HER2＋ lesions.
文摘11C-labeled C1-C10 partial structure of kulokekahilide-2 (1) was successfully synthesized based on Pd0-mediated rapid C-[11C]methylation using [11C]methyl iodide and pinacol alkenylboronate. The preparation of organoboron intermediate via olefin cross-metathesis is also a crucial procedure for the synthesis of 11C-labeling C1-C10 dihy-droxy acid moiety of 1.
文摘A k-L(2,1)-labeling for a graph G is a function such that whenever and whenever u and v are at distance two apart. The λ-number for G, denoted by λ(G), is the minimum k over all k-L(2,1)-labelings of G. In this paper, we show that for or 11, which confirms Conjecture 6.1 stated in [X. Li, V. Mak-Hau, S. Zhou, The L(2,1)-labelling problem for cubic Cayley graphs on dihedral groups, J. Comb. Optim. (2013) 25: 716-736] in the case when or 11. Moreover, we show that? if 1) either (mod 6), m is odd, r = 3, or 2) (mod 3), m is even (mod 2), r = 0.
基金supported by Beijing Natural Science Foundation(No.7132037)the National Cancer Institute Network for Translational Research U54 CA136404-01
文摘Objective： To investigate the effect of fluorescent dye labeling on the targeting capabilities of 111 In- （DTPA）.-trastuzumab-（IRDye 800）m. Methods： Trastuzumab-based conjugates were synthesized and conjugated with diethylenetriaminepentaacetic acid （DTPA） at molar ratios of I, 2, 3 and 5 and with a fluorescent dye （IRDye 800CW） at molar ratios of 1, 3 and 5. Immunoreactivity and internalization were assessed on SKBR-3 cells, overexpressing human epidermal growth factor receptor 2. The stability in human serum and phosphate-buffered saline （PBS） was evaluated. The biodistribution of dual-labeled conjugates was compared with that of 111In-（DTPA）2-trastuzumab in a SKBR-3 xenograft model to evaluate the effect of dye-to-protein ratio. Results： All trastuzumab-based conjugates exhibited a high level of chemical and optical purity. Flow cytometry results showed that increasing dye-to-protein ratios were associated with decreased immunoreactivity. Stability studies revealed that the conjugate was stable in PBS, while in human serum, increased degradation and protein precipitation were observed with increasing dye-to-protein ratios. At 4 h, the percentages of internalization of dual-labeled conjugates normalized by dye-to-protein ratio （m） were 24.88%±2.10%, 19.99%±0.59%, and 17.47%±1.26% for ＂m＂ equal to 1, 3, and 5, respectively. A biodistribution study revealed a progressive decrease in tumor uptake with an increase in the dye-to-protein ratios. The liver, spleen and kidney showed a marked uptake with increased dye-to-protein ratios, particularly in the latter. Conclusions： With non-specific-site conjugation of the fluorescent dye with a protein based on imaging agent, the increase in dye-to-protein ratios negatively impacted the immunoreactivity and stability, indicating a reduced tumor uptake.
文摘the widely discussed study of word class categorization has a long history of more than 2000 years,which is known as the study of the“God Particles”in language.As a typical analytic language,Modern Chinese,due to its lack of morphological changes,is challenged by a thorny problem of word classes especially when it comes to the criteria for word class identification and the treat⁃ment of multiple class membership.As such,all the controversies eventually give rise to some contradiction and confusion in word class labeling in Modern Chinese and Chinese-English dictionaries.As an important grammatical means in Chinese and the focus of lexicology and rhetorics,total reduplication lexemes serve as an essential part of Chinese-English dictionaries with complex and diverse word classes.Guided by the Two-level Word Class Categorization Theory,this thesis focuses on the word class labeling of total reduplication lexemes in New Century Chinese-English Dictionary(2nd edition)backed by large-scale balanced Modern Chi⁃nese corpora.With an innovative theoretical perspective,this study not only contributes to the word class labeling of total reduplica⁃tion lexemes and even sheds light on the compilation of Chinese-English dictionaries,but also drives the study of Modern Chinese word classes in the long term.
