Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.M...Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods.展开更多
Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in ...Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity.展开更多
Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% so...Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% sodium periodate and 5% chromic acid were used as oxidant respectively. Theoxidation time for the mouse lung smears was 5,15,60 minutes and the oxidation temperature was 20℃.The time of silver impregnation was 90 minutcs and the temperature was 60℃ for the all smearo. Whenthe oxidation time was under 15 minutes. Pneumocystis cariniic cysts showed light or dark brown, and theparenthesis-like structure could clearly be found in part of the cysts. However, if the time of oxidationWas longer, the cysts showed black and secmed to have damaged. In the same batch of the mouse lungsmears oxidated for 5 minutes, the samiples oxidated by sodium periodate showed more the cysts with theparen thesis-like structure than those oxidated by chromic acid.In the rat or patient's lung sectionsoxidated by. sodium periodate, this structure could also be found. The result of the experiment showsthat sodium periodate as an oxidant in the subsequent step of the the silver impregnation is preferable tochromic acid. And then,it is useful to clinical practice that the step of sodium bisulfate can be omittedin the study.展开更多
in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue secti...in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue sections or cell preparations. Probes are usually展开更多
Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize...Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize root tips using immunocytochemistry technique.Based on the staining pattern ob-served along the root tip,an apicobasal gradient of ABBP has been revealed with either Ab Ⅰ or Ab Ⅱ.Thelabels are mainly concentrated in,the root cap and apical meristem.In the elongation zone and root hair zone,only cells in pericycle and epidermis are obviously labeled.The silver labels are distributed evenly in thewhole cell of the apical meristem,but in the cells of the root cap and elongation zone,the positive stainingdegree in the cytoplasm is reduced even to completely negative staining;nevertheless,the nuclei and plasmamembranes are strongly labeled.As in the labeled cells of the root hair zone,only some immunoactive sitescan be seen in the nucleus compacted to the edge of the cells.The staining pattern in the whole root tip alsoshows obvious dependence of the ABBPs localization on the activity of the nucleus.The significance ofABBPs localization is discussed.展开更多
Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of tre...Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of treating hypersensitivity for individuals with chronic teeth pain. SDF is also used as a method to treat and arrest dental caries. SDF application is limited due to its negative esthetic effects, which is a black stain where the cavity was present on the tooth. Topical application of potassium iodide applied immediately after SDF has been shown in studies to reduce the color change caused by SDF. This study used topical application of silver diamine fluoride (SDF) and potassium iodide (KI) treatments on bovine teeth to determine if SDF and KI were efficacious in the treatment for carious lesions. The color change was detected by use of spectrophotometric analysis to determine L, a and b readings that demarcate light and color values following staining. The conclusion was made that the application of SDF followed directly by KI treatment produced L, a and b spectrophotometric values that indicated a significant reduction in teeth staining than the application of SDF alone. Therefore, this study supports the idea that SDF and KI can be used to treat carious lesions on bovine teeth while retaining surface enamel coloration.展开更多
[Objective] The aim was to optimize the SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Method]25 accessions of kernelled apricot and three accessions of edible apricot were s...[Objective] The aim was to optimize the SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Method]25 accessions of kernelled apricot and three accessions of edible apricot were selected as experimental materials to screen the repeatable SSR loci with high polymorphism by the use of SSR markers combined with non-denatured polyacrylamide gel electrophoresis.And the effect of different factors on electrophoresis conditions was compared to explore the optimal SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Result]The optimal non-denatured polyacrylamide gel electrophoresis conditions for SSR-PCR were established as follows：polyacrylamide gel concentration 6%,the ratio of acrylamide to bisacrylamide 29∶1,electrophoresis at 1 000 V for 2-3 h,and staining for 15 min within 0.1% AgNO3.[Conclusion]The optimum electrophoresis system has provided some technical foundations to further study the phylogenetic relationship of kernelled apricots by SSR markers.展开更多
[Objective] The aim was to explore biological function of related genes in Setcreasea purpurea Boom under copper stress and to study the mechanism of its copper-resistance from standpoint of molecular biology in order...[Objective] The aim was to explore biological function of related genes in Setcreasea purpurea Boom under copper stress and to study the mechanism of its copper-resistance from standpoint of molecular biology in order to solve the problem of copper pollution. [Method] Setcreasea purpurea Boom was taken as experimental material which enjoys enrichment ability to cupric ion. About 90 fragments of differential expression were obtained by cDNA-AFLP and silver staining technique, among which, 17 fragments were amplified. After purification and identification, sequences of 6 differential fragments were got and used for BLAST X contrast. [Result] Six differential expressed fragments may play roles when Setcreasea purpurea Boom was under copper stress. The homology achieved 49% between differential sequences of E5MG-3 and of Arabidopsis thaliana mRNA （accession numbers： AAM62956.1）, homology was 53% between sequences of E4MB-2 and Solanum tuberosum mRNA （accession numbers： A5A717.1）, and 65% between sequences of E6MG-1 and Colocasia esculenta （L.） Schott mRNA （accession numbers： AAO62313.1）. It can be concluded that differential expressed genes are related to cell signaling, antioxidation, metabolism and protein modification. [Conclusion] The study has laid foundation for further exploration of regulatory network about response of Setcreasea purpurea Boom to copper stress.展开更多
Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of ...Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin （STZ）. At the second week after STZ injection, the sensory nerve conduction velocities （SNCV） of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction （DD-PCR） was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, （45.25±10.38） m/s] reduced obviously compared with the control group [n = 8, （60.10± 11.92） m/s] （P 〈 0.05）. Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA （accession No., BC059140）, which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.展开更多
基金Supported by the 2016 Major Collaborative Innovation Program of the Chinese Academy of Medical Sciences(2016-I2M-1004)
文摘Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods.
文摘Telomerase is a ribonucleoprotein enzyme, which synthesizes telomeric repeats (TTAGGG) n.While germline cells and most malignant tumor cells express telomerase activity, normal somatic cells aregenerally deficient in telomerase activity. Our objective was to detect telomerase activity of human cells bysilver staining with telorneric repeat amplification protocol (TRAP) which is easy and quick. Comparing withradioisotopic TRAP, we examined the telomerase activity in telomerase-positive 293-cell and RNase-pretreated and heat-pretreated negative controls by silver staining TRAP. We detected telomerase activity in 2 strainsof human liver tumor cells (QGY7701 and SMMC7721 ). The 293 cells (only 10 cells) and the 2 strains ofhuman liver tumor cells were all positive. while telomerase activity was not detected in the negative controls.These data suggest that non-radioisotopic silver staining TRAP is a specific, sensitive and fast assay fortelomerase activity. It was verified that the 2 strains of human liver tumor cells express telomerase activity.
文摘Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% sodium periodate and 5% chromic acid were used as oxidant respectively. Theoxidation time for the mouse lung smears was 5,15,60 minutes and the oxidation temperature was 20℃.The time of silver impregnation was 90 minutcs and the temperature was 60℃ for the all smearo. Whenthe oxidation time was under 15 minutes. Pneumocystis cariniic cysts showed light or dark brown, and theparenthesis-like structure could clearly be found in part of the cysts. However, if the time of oxidationWas longer, the cysts showed black and secmed to have damaged. In the same batch of the mouse lungsmears oxidated for 5 minutes, the samiples oxidated by sodium periodate showed more the cysts with theparen thesis-like structure than those oxidated by chromic acid.In the rat or patient's lung sectionsoxidated by. sodium periodate, this structure could also be found. The result of the experiment showsthat sodium periodate as an oxidant in the subsequent step of the the silver impregnation is preferable tochromic acid. And then,it is useful to clinical practice that the step of sodium bisulfate can be omittedin the study.
文摘in situ hybridization or hybridocytochemistry is a powerful tool for detection and localization of gene expression. It is also the most commonly used procedure to localize specific DNA or RND sequences in tissue sections or cell preparations. Probes are usually
文摘Antisera against abseisic-acid-binding proteins(Ab Ⅰ)and anti-idiotypic antibodies to ABAmonoclonal antibody(Ab Ⅱ)have been developed and tested to comparatively localize abacisic-acid-bindingproteins(ABBPs)in maize root tips using immunocytochemistry technique.Based on the staining pattern ob-served along the root tip,an apicobasal gradient of ABBP has been revealed with either Ab Ⅰ or Ab Ⅱ.Thelabels are mainly concentrated in,the root cap and apical meristem.In the elongation zone and root hair zone,only cells in pericycle and epidermis are obviously labeled.The silver labels are distributed evenly in thewhole cell of the apical meristem,but in the cells of the root cap and elongation zone,the positive stainingdegree in the cytoplasm is reduced even to completely negative staining;nevertheless,the nuclei and plasmamembranes are strongly labeled.As in the labeled cells of the root hair zone,only some immunoactive sitescan be seen in the nucleus compacted to the edge of the cells.The staining pattern in the whole root tip alsoshows obvious dependence of the ABBPs localization on the activity of the nucleus.The significance ofABBPs localization is discussed.
文摘Silver Diamine Fluoride (SDF) is colorless and alkaline with a pH of 10. It has been used in Japan and other international countries for decades. The Food and Drug Administration gave approval for it as a means of treating hypersensitivity for individuals with chronic teeth pain. SDF is also used as a method to treat and arrest dental caries. SDF application is limited due to its negative esthetic effects, which is a black stain where the cavity was present on the tooth. Topical application of potassium iodide applied immediately after SDF has been shown in studies to reduce the color change caused by SDF. This study used topical application of silver diamine fluoride (SDF) and potassium iodide (KI) treatments on bovine teeth to determine if SDF and KI were efficacious in the treatment for carious lesions. The color change was detected by use of spectrophotometric analysis to determine L, a and b readings that demarcate light and color values following staining. The conclusion was made that the application of SDF followed directly by KI treatment produced L, a and b spectrophotometric values that indicated a significant reduction in teeth staining than the application of SDF alone. Therefore, this study supports the idea that SDF and KI can be used to treat carious lesions on bovine teeth while retaining surface enamel coloration.
基金Supported by the Scientific and Technological Program of Educational Department of Hebei Province(No.ZH2007116)~~
文摘[Objective] The aim was to optimize the SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Method]25 accessions of kernelled apricot and three accessions of edible apricot were selected as experimental materials to screen the repeatable SSR loci with high polymorphism by the use of SSR markers combined with non-denatured polyacrylamide gel electrophoresis.And the effect of different factors on electrophoresis conditions was compared to explore the optimal SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Result]The optimal non-denatured polyacrylamide gel electrophoresis conditions for SSR-PCR were established as follows：polyacrylamide gel concentration 6%,the ratio of acrylamide to bisacrylamide 29∶1,electrophoresis at 1 000 V for 2-3 h,and staining for 15 min within 0.1% AgNO3.[Conclusion]The optimum electrophoresis system has provided some technical foundations to further study the phylogenetic relationship of kernelled apricots by SSR markers.
基金Supported by National Natural Science Foundation of China(30760021)Natural Science Foundation of Jiangxi Province(0530016)Project from Education Bureau of Jiangxi Province([2007]151)~~
文摘[Objective] The aim was to explore biological function of related genes in Setcreasea purpurea Boom under copper stress and to study the mechanism of its copper-resistance from standpoint of molecular biology in order to solve the problem of copper pollution. [Method] Setcreasea purpurea Boom was taken as experimental material which enjoys enrichment ability to cupric ion. About 90 fragments of differential expression were obtained by cDNA-AFLP and silver staining technique, among which, 17 fragments were amplified. After purification and identification, sequences of 6 differential fragments were got and used for BLAST X contrast. [Result] Six differential expressed fragments may play roles when Setcreasea purpurea Boom was under copper stress. The homology achieved 49% between differential sequences of E5MG-3 and of Arabidopsis thaliana mRNA （accession numbers： AAM62956.1）, homology was 53% between sequences of E4MB-2 and Solanum tuberosum mRNA （accession numbers： A5A717.1）, and 65% between sequences of E6MG-1 and Colocasia esculenta （L.） Schott mRNA （accession numbers： AAO62313.1）. It can be concluded that differential expressed genes are related to cell signaling, antioxidation, metabolism and protein modification. [Conclusion] The study has laid foundation for further exploration of regulatory network about response of Setcreasea purpurea Boom to copper stress.
基金the grant from Technical Program of Social Development ofNantong Municipality (No.S30043)the Natural ScienceFoundation of Nantong University (No. 05Z084)
文摘Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin （STZ）. At the second week after STZ injection, the sensory nerve conduction velocities （SNCV） of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction （DD-PCR） was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, （45.25±10.38） m/s] reduced obviously compared with the control group [n = 8, （60.10± 11.92） m/s] （P 〈 0.05）. Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA （accession No., BC059140）, which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.
