Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before...Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.展开更多
INTRODUCTION The hepatitis A virus specific immunoglobulin M(IgM)antibody is a specific serological marker forearly diagnosis of hepatitis A..At present,themethods used at home or abroad for detecting anti-HAV IgM are...INTRODUCTION The hepatitis A virus specific immunoglobulin M(IgM)antibody is a specific serological marker forearly diagnosis of hepatitis A..At present,themethods used at home or abroad for detecting anti-HAV IgM are RIA,ELISA and SPHAI.The dotimmunogold combination assay that has beendeveloped since 1989 is a new technique with theproperty of simple and rapid immunologicaldetection,by using the red colloidal gold particles tolabel the antibodies as indicator,and the展开更多
AIM To detect hepatitis A virus-specific immunoglobulin M (IgM) antibody rapidly.METHODS Colloidal gold with an average dia-meter of 15 nm was prepared by controlled reduction of a boiling solution of 0.2 g/ L chloroa...AIM To detect hepatitis A virus-specific immunoglobulin M (IgM) antibody rapidly.METHODS Colloidal gold with an average dia-meter of 15 nm was prepared by controlled reduction of a boiling solution of 0.2 g/ L chloroauric acid with 10 g/ L sodium citrate and labeled with anti-HAVIgG as gold probe. Dot immunogold filtration assay (DIGFA) has been developed by coating anti-human μ chain on nitrocellulose membrane (NCM) for capturing the anti-HAV IgM in serum, then using cultured hepatitis A antigen as a 'bridge', connecting anti-HAV IgM in sample and anti-HAV IgG labeled colloidal gold. If there was anti-HAV IgM in sample, gold probes would concentrate on NCM, which will appear a pink dot.RESULTS A total of 264 serum samples were comparatively detected with both DIGFA and ELISA by 'blind' method. Among them, 88 were positive and 146 were negative with the two methods. The sensitivity and the specificity of DIGFA were 86.27% and 90.12%, respectively. Fifteen negative serum samples and 15 positive serum samples were detected 3 times repeatedly, the results were the same.CONCLUSION DIGFA is a simple, rapid, sensitive, specific and reliable method without expensive equipment and is not interfered with rheumatoid factor (RF) in serum. It is suitable for basic medical laboratories. The test could be applied for diagnosis and epidemiological survey of hepatitis A. It has a broad prospect in application.INTRODUCTIONHepatitis A virus-specific immunoglobulin M antibody (anti-HAV IgM) is the specific serological marker for the early diagnosis of acute hepatitis A. It can be detected by radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), solid phase hemagglutination inhibition test (SPHIA) and other methods. At present, double sandwich ELISA is in widespread use[1]. However, it takes more time to finish the test and the procedure is complicated. The need of a simple, rapid, and noninstrumented test is evident in many basic units, where laboratory facilities and trained personnel are limited. In 1989, Chun developed a rapid test, DIGFA[2]. It has been used to detect HCG, C-reactive protein, immunoglobulin G antibody and others[3,4]. We developed DIGFA for detection of anti-HAV IgM. The evaluation of this test is presented below.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
Apolipoprotein A-IMilano(ApoA-IM)has been shown to significantly reduce coronary atherosclerotic plaques.However,the preparation of cost-effective pharmaceutical formulations of ApoA-IM is limited by the high cost and...Apolipoprotein A-IMilano(ApoA-IM)has been shown to significantly reduce coronary atherosclerotic plaques.However,the preparation of cost-effective pharmaceutical formulations of ApoA-IM is limited by the high cost and difficulty of purifying the protein and producing the highly effective dimeric form.The aim of this study was to create an expression cassette that specifically drives the expression of dimeric ApoA-IM in the protein bodies of rice seeds.The ApoA-IM protein under control of the 13 kDa prolamin promoter is expressed exclusively in its dimeric form within the seeds,and immunocytochemical and immunogold analyses confirmed its expression in different caryopsis tissue such as seed coat,aleurone cell and endosperm,particularly in amyloplast and storage vacuoles.A plant-based ApoA-IM production system offered numerous advantages over current production systems,including the direct production of the most therapeutically effective dimeric ApoA-IM forms,long-term protein storage in seeds,and ease of protein production by simply growing plants.Therefore,seeds had the potential to serve as a costeffective source of therapeutic ApoA-IM.展开更多
AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmun...AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmune process of bile duct destruction in primary biliary cirrhosis（PBC）．METHODS：Ten wedged liver biopsies of PBC（five cases each of stages 2 and 3）were studied. The liver specimens were processed for transmission electron microscopy. Immunohistochemistry was performed using anti-ICAM-1 and anti-LFA-1 mouse mAbs.In situ hybridization was done to examine the messenger RNA expression of ICAM-1 in formalin-fixed．paraffin-embedded sections using peptide nucleic acid probes and the catalyzed signal amplification （CSA）technique．Immunogold-silver staining for electron microscopy was Derrormed using anti-ICAM and anti-LFA-1 mouse mAbs．The immunogold particles on epithelial cells of bileductules and cholangiocytes of CoH cells were counted and analyzed semi-quantitatively．Western blotting was performed to confirm ICAM-1 protein expression．RESULTS：In liver tissues of PBC patients．immunohi-stochemistry showed aberrant ICAM-1 expression on the plasma membrane of epithelial cells lining bile ductules，and also on mature cholangiocytes but not on hepatocytes in CoH．LFA-1-positive lymphocytes were closely associated with epithelial cells in bile ductules．ICAM-1 expression at protein level was confirmed by Western blot．In situ hybridization demonstrated ICAM-1 mRNA expression in bile ductules and LFA-1 mRNA in lymphocytes infiltrating the bileductules．By immunoelectron microscopy，ICAM-1 was demonstrated on the basal suface of epithelial cells in bile ductules and on the luminal surfaces of cholangiocytes in damaged COH．Cells with intermediate morphology resembling progenitor cells in Coil were not labeled with ICAM-1 and LFA-1．CONCLUSION：De novo expression of ICAM-1 both on mature cholangiocytes in COH and epithelial cells in bile ductules in PBC implies that lymphocyte-induced destruction through adhesion by ICAM-1 and binding of LFA-1-expressing activated lymphocytes takes place not only in bile ductules but also in the COH．展开更多
文摘Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.
文摘INTRODUCTION The hepatitis A virus specific immunoglobulin M(IgM)antibody is a specific serological marker forearly diagnosis of hepatitis A..At present,themethods used at home or abroad for detecting anti-HAV IgM are RIA,ELISA and SPHAI.The dotimmunogold combination assay that has beendeveloped since 1989 is a new technique with theproperty of simple and rapid immunologicaldetection,by using the red colloidal gold particles tolabel the antibodies as indicator,and the
文摘AIM To detect hepatitis A virus-specific immunoglobulin M (IgM) antibody rapidly.METHODS Colloidal gold with an average dia-meter of 15 nm was prepared by controlled reduction of a boiling solution of 0.2 g/ L chloroauric acid with 10 g/ L sodium citrate and labeled with anti-HAVIgG as gold probe. Dot immunogold filtration assay (DIGFA) has been developed by coating anti-human μ chain on nitrocellulose membrane (NCM) for capturing the anti-HAV IgM in serum, then using cultured hepatitis A antigen as a 'bridge', connecting anti-HAV IgM in sample and anti-HAV IgG labeled colloidal gold. If there was anti-HAV IgM in sample, gold probes would concentrate on NCM, which will appear a pink dot.RESULTS A total of 264 serum samples were comparatively detected with both DIGFA and ELISA by 'blind' method. Among them, 88 were positive and 146 were negative with the two methods. The sensitivity and the specificity of DIGFA were 86.27% and 90.12%, respectively. Fifteen negative serum samples and 15 positive serum samples were detected 3 times repeatedly, the results were the same.CONCLUSION DIGFA is a simple, rapid, sensitive, specific and reliable method without expensive equipment and is not interfered with rheumatoid factor (RF) in serum. It is suitable for basic medical laboratories. The test could be applied for diagnosis and epidemiological survey of hepatitis A. It has a broad prospect in application.INTRODUCTIONHepatitis A virus-specific immunoglobulin M antibody (anti-HAV IgM) is the specific serological marker for the early diagnosis of acute hepatitis A. It can be detected by radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), solid phase hemagglutination inhibition test (SPHIA) and other methods. At present, double sandwich ELISA is in widespread use[1]. However, it takes more time to finish the test and the procedure is complicated. The need of a simple, rapid, and noninstrumented test is evident in many basic units, where laboratory facilities and trained personnel are limited. In 1989, Chun developed a rapid test, DIGFA[2]. It has been used to detect HCG, C-reactive protein, immunoglobulin G antibody and others[3,4]. We developed DIGFA for detection of anti-HAV IgM. The evaluation of this test is presented below.
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Apolipoprotein A-IMilano(ApoA-IM)has been shown to significantly reduce coronary atherosclerotic plaques.However,the preparation of cost-effective pharmaceutical formulations of ApoA-IM is limited by the high cost and difficulty of purifying the protein and producing the highly effective dimeric form.The aim of this study was to create an expression cassette that specifically drives the expression of dimeric ApoA-IM in the protein bodies of rice seeds.The ApoA-IM protein under control of the 13 kDa prolamin promoter is expressed exclusively in its dimeric form within the seeds,and immunocytochemical and immunogold analyses confirmed its expression in different caryopsis tissue such as seed coat,aleurone cell and endosperm,particularly in amyloplast and storage vacuoles.A plant-based ApoA-IM production system offered numerous advantages over current production systems,including the direct production of the most therapeutically effective dimeric ApoA-IM forms,long-term protein storage in seeds,and ease of protein production by simply growing plants.Therefore,seeds had the potential to serve as a costeffective source of therapeutic ApoA-IM.
文摘AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmune process of bile duct destruction in primary biliary cirrhosis（PBC）．METHODS：Ten wedged liver biopsies of PBC（five cases each of stages 2 and 3）were studied. The liver specimens were processed for transmission electron microscopy. Immunohistochemistry was performed using anti-ICAM-1 and anti-LFA-1 mouse mAbs.In situ hybridization was done to examine the messenger RNA expression of ICAM-1 in formalin-fixed．paraffin-embedded sections using peptide nucleic acid probes and the catalyzed signal amplification （CSA）technique．Immunogold-silver staining for electron microscopy was Derrormed using anti-ICAM and anti-LFA-1 mouse mAbs．The immunogold particles on epithelial cells of bileductules and cholangiocytes of CoH cells were counted and analyzed semi-quantitatively．Western blotting was performed to confirm ICAM-1 protein expression．RESULTS：In liver tissues of PBC patients．immunohi-stochemistry showed aberrant ICAM-1 expression on the plasma membrane of epithelial cells lining bile ductules，and also on mature cholangiocytes but not on hepatocytes in CoH．LFA-1-positive lymphocytes were closely associated with epithelial cells in bile ductules．ICAM-1 expression at protein level was confirmed by Western blot．In situ hybridization demonstrated ICAM-1 mRNA expression in bile ductules and LFA-1 mRNA in lymphocytes infiltrating the bileductules．By immunoelectron microscopy，ICAM-1 was demonstrated on the basal suface of epithelial cells in bile ductules and on the luminal surfaces of cholangiocytes in damaged COH．Cells with intermediate morphology resembling progenitor cells in Coil were not labeled with ICAM-1 and LFA-1．CONCLUSION：De novo expression of ICAM-1 both on mature cholangiocytes in COH and epithelial cells in bile ductules in PBC implies that lymphocyte-induced destruction through adhesion by ICAM-1 and binding of LFA-1-expressing activated lymphocytes takes place not only in bile ductules but also in the COH．
