ABSTRACT
Objective To investigate the difference between mammary gland tissues and breast cancer tissues.Methods Monoclonal antibodies against Mam-A immunized epitopes were screened for immunohistochemical staining of normal breast tissues and breast cancer tissues.The average optical density was used as an index to identify the quantitative data by computer-aided technology to screen epitope-specific antibodies with significant difference in staining characteristics between two types of tissues.Furthermore the feasibility and effectiveness of breast cancer diagnosis were evaluated.Results Four anti-Mam-A epitope-specific monoclonal antibodies,mAb1152,mAb11617,mAb995 and mAb656,were obtained.Immunohistochemical staining showed that the average density of mAb1152,mAb11617 and mAb995 was significantly different between the two types of tissues.The difference was significant between normal breast tissues and breast cancer tissues under the same conditions.The results showed that mAb11617 was better than mAb1152 and mAb995.At the best working point,mAb11617 was the best,the specificity was 90％ and the sensitivity was 59.62％.Further analysis showed that the sensitivity of mAb11617 combined with mAb995 in the diagnosis of in situ breast cancer was 81.48％ and the specificity was 90％,which was of great diagnostic significance.Conclusion There is significant difference between breast tissues and breast cancer tissues in Mam-A protein immunological activity or expression.This difference,which can be recognized by the specific antibody staining and computer aided technology,is of important diagnostic value.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of Bushen Qiangji Granule (, BSQJ) in restraining the osteogenic differentiation of ankylosing spondylitis (AS) fifibroblasts.</p><p><b>METHODS</b>Hip joint capsules were obtained from AS patients (n=10) receiving total hip replacement and healthy hip joint capsules from patients with hip fracture (n=10) receiving surgery as a control. Finite fifibroblast lines were established from these tissue samples to observe the effect of BSQJ on suppressing osteogenic differentiation of fifibroblasts. The expression of osteogenic marker gene corebinding factor a1 (Cbfa1) and Smad family proteins were examined by Western blot and real-time quantitative polymerase chain reaction (qPCR).</p><p><b>RESULTS</b>The mRNA expression level of Cbfa1 was significantly higher in AS fibroblasts than that in normal fibroblasts and the expression of pSmad1, pSmad5, Smad4 and Cbfa1 in AS fibroblasts was also higher, demonstrating the activation of the BMP/Smads signal pathway in AS fifibroblasts. BSQJ-medicated serum not only restrained the mRNA and protein expression levels of Cbfa1 and inhibited protein expression level of Smad4 but also decreased the expression quantities of pSmad1 and pSmad5.</p><p><b>CONCLUSIONS</b>BSQJ can inhibit osteogenic differentiation of AS fifibroblasts in vitro by suppressing the activation of the BMP/Smads signal pathway. This may be the important molecular mechanism of BSQJ in regulating AS ossifification.</p>
Subject(s)
Adult , Humans , Middle Aged , Young Adult , Bone Morphogenetic Proteins , Metabolism , Cell Differentiation , Core Binding Factor Alpha 1 Subunit , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Fibroblasts , Metabolism , Pathology , Osteogenesis , Genetics , Phosphorylation , RNA, Messenger , Genetics , Metabolism , Serum , Metabolism , Signal Transduction , Smad Proteins , Metabolism , Spondylitis, Ankylosing , Genetics , PathologyABSTRACT
<p><b>OBJECTIVE</b>To study the pulmonary pathology in patients died of fatal human influenza A(H1N1) infection.</p><p><b>METHODS</b>Eight cases of fatal human influenza A (H1N1) infection, including 2 autopsy cases and 6 paramortem needle puncture biopsies, were enrolled into the study. Histologic examination, immunohistochemitry, flow cytometry and Western blotting were carried out.</p><p><b>RESULTS</b>The major pathologic changes included necrotizing bronchiolitis with surrounding inflammation, diffuse alveolar damage and pulmonary hemorrhage. Influenza viral antigen expression was detected in the lung tissue by Western blotting. Immunohistochemical study demonstrated the presence of nuclear protein and hemagglutinin virus antigens in parts of trachea, bronchial epithelium and glands, alveolar epithelium, macrophages and endothelium. Flow cytometry showed that the apoptotic rate of type II pneumocytes (32.15%, 78.15%) was significantly higher than that of the controls (1.93%, 3.77%).</p><p><b>CONCLUSION</b>Necrotizing bronchiolitis, diffuse alveolar damage and pulmonary hemorrhage followed by pulmonary fibrosis in late stage are the major pathologic changes in fatal human influenza A (H1N1) infection.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Alveolar Epithelial Cells , Pathology , Antigens, Viral , Metabolism , Apoptosis , Autopsy , Biopsy, Needle , Bronchiolitis, Viral , Pathology , Hemagglutinin Glycoproteins, Influenza Virus , Metabolism , Influenza A Virus, H1N1 Subtype , Allergy and Immunology , Influenza, Human , Metabolism , Mortality , Pathology , Virology , Lung , Allergy and Immunology , Metabolism , Pathology , Nuclear Proteins , Metabolism , Pulmonary Alveoli , Pathology , Pulmonary Fibrosis , PathologyABSTRACT
<p><b>OBJECTIVE</b>To detect humoral immune response against different function regions of hepatitis C virus (HCV) in chronic patients, and further to investigate the correlativity between anti-HCV antibody titers and HCV RNA concentration.</p><p><b>METHODS</b>Using recombinant dominate epitope antigens, e.g. HCV Core, NS3, NS4, NS5 and chimeric HVR1, a set of ELISA test reagents was formulated. Then, titers of antibodies against HCV different regions and the RNA concentration of HCV in chronic patient sera were detected by ELISA and quantitative RT-PCR technique, respectively.</p><p><b>RESULTS</b>Great differences have been noted in antibody titers and positive rate of different HCV function regions in chronic patients. Antibodies against HCV Core and HVR1 have the highest positive rate, then NS3, NS4, and NS5 in sequence.</p><p><b>CONCLUSION</b>The titer of antibodies against different regions of HCV in chronic patients has good correlation with HCV RNA concentration.</p>
Subject(s)
Humans , Hepatitis C Antibodies , Blood , Hepatitis C, Chronic , Allergy and Immunology , Virology , RNA, Viral , BloodABSTRACT
Objective: To study the recombinant epitope antigens of hepatitis C virus (HCV), in order to fulfil the requirements of recombinant immunoblot assay kit. Methods: An expressing vector pBVIL1 for expression of recombinant antigens in a fusion manner with IL-1β was constructed. A series of selected genes from the HCV antigens including the C, NS3, NS4 and NS5 were amplified from HCV gene-containing plasmids using PCR and the expression plasmids for these genes were constructed in pBVIL1, respectively. The activity of the purified recombinant antigens were tested against an identified HCV antibody positive and negative panel with ELISA. Results and Conclusions: All the cloned genes of chosen antigen epitopes were highly expressed in pBVIL1 in E.coli. The activity of the C and NS4 antigens were slightly higher than the RIBA3.0 antigens, while the activity of NS3 was slightly lower than the RIBA3.0 antigen. But the total evaluation for the panel was same as RIBA3.0. That means the cloned antigens were suitable for the use in RIBA test kit.