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Objective To explore the significance of combined detection of urinary podocalyxin (PCX), nephrin and mono-cyte chemoattractant protein-1(MCP-1) for diabetic nephropathy in newly diagnosed type 2 diabetic patients.Methods A total of 132 newly diagnosed type 2 diabetes mellitus (T2DM) patients and 36 healthy subjects were recruited .The concentrations of urinary PCX (UPCX), nephrin (UNephrin) and MCP-1(UMCP-1) were measured with enzyme-linked immunosorbent assay (ELISA), and urina-ry UAE ( UAlb) concentration was measured with turbidimetric immunoassay method .All the results were shown by dividing the urine creatinine ( UCR) concentration in the same sample .The comparisons among UAlb/UCR, UPCX/UCR, UNephrin/UCR, and UMCP-1/UCR among groups were investigated .The correlation among UAlb/UCR, UPCX/UCR, UNephrin/UCR and UMCP-1/UCR was in-vestigated in T2DM patients.The comparison among positive rate of UMCP-1/UCR, UNephrin/UCR, UPCX/UCR and UAlb/UCR was investigated in T2DM patients.Results The levels of UPCX/UCR and UMCP-1/UCR were significantly higher in diabetes melli-tus(DM) group (37.37 ±22.83 and 47.19 ±29.98, respectively) than in healthy subjects (9.59 ±2.28 and 9.98 ±3.63, respec-tively) ( P <0.05 ) .The levels of UAlb/UCR、UPCX/UCR and UMCP-1/UCR were significantly higher in diabetic nephropathy (DN) group (90.39 ±59.35, 72.11 ±58.88, and 104.82 ±82.77, respectively) than in DM group (17.50 ±9.49, 37.37 ± 22.83 , and 47.19 ±29.98 , respectively ) and in healthy subjects ( 8.93 ±3.81 , 9.59 ±2.28 , and 9.98 ±3.63 , respectively ) ( P<0.05).The level of UNephrin/UCR was significantly higher in DN group (83.60 ±61.61) than in DM group (38.74 ±35.27) ( P <0.05).The levels of UPCX/UCR, UMCP-1/UCR, and UNephrin/UCR were positively correlated with UAlb/UCR ( r =0.619, 0.568, and 0.535, respectively) ( P <0.05) in T2DM patients.The level of UPCX/UCR was positively correlated with the levels of UMCP-1/UCR and UNephrin/UCR ( r =0.799 and 0.718 , respectively ) ( P <0.05 ) .The level of UNephrin/UCR was positively correlated with the level of UMCP-1/UCR ( r =0.635 P <0.05) in T2DM patients.The positive rate of UMCP-1/UCR, UNephrin/UCR, and UPCX/UCR was significantly higher than that of UAlb/UCR(χ2 =9.818, 6.673, and 5.395, respectively, P <0.05)in T2DM patients.Conclusions The combined detection of urinary PCX , Nephrin and MCP-1 is very important for the early renal dam-age in newly diagnosed type 2 diabetic patients .
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ObjectiveTo observe the expression of bone sialoprotein(BSP) and matrix metalloproteinase-9 (MMP-9) in calcified valves of patients with rheumatic heart disease.MethodsA total of 150 mitral valves which were resected by surgery were divided into rheumatic group ( 120 valves) and nonrheumatic group (30 valves).Immunohistochemical staining was taken by SP method and the expressions of BSP and MMP-9 in two groups were observed and compared.ResultsThe positive expressions of BSP and MMP-9 in rheumatic group were 91.7％(110/120) and 90.8％(109/120),respectively,which were significantly higher than those in non-rheumatic group [23.3％(7/30) and 20.0％(6/30) ](P＜ 0.01 ).Conclusions The expressions of both BSP and MMP-9 are higher in the valves of patients with rheumatic heart disease.The calcification of rheumatic mitral valves is closely related with the degradation and remodeling of extracellular matrix caused by MMP-9,and osteoblast-like bone formation induced by BSP.
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Objective The study was aiming at elucidating the rale of BSP over-expression in enhancing osseous metastasis of breast cancer cells,in order to search for new drug targets for prevention and treatment of such metastasis.Methods Chicken embryo allantois membrane(CAM)was used in the experiment.Breast cancer cells were inoculated on the superior surface of CAM sac,then the vascular tissues were collected in lower sac after incubation.The human specific Alu gene was amplified by PCR with genomic DNA extracted as template.Breast cancer cells were incubated with bone,which had becn treated with collagenase and trypsinase,and then OD_(488) of breast cancer was assessed to evaluate the adhesive affinity of breast cancer cells to bone.Results The findings of CAM assay and PCR amplifica- tion using human Alu primers and genomic DNA showed that Alu gene was amplified from 4 of 5 in MDA-MB-231BO(pI)group and 3 of 5 in MDA-MB-231BO(pID)group,but not amplified in control group.In bone adhesion assay,the OD_(488) of digested bone after MDA- MB-231BO(pI)and MDA-MB-231BO(pID)adhering to bone were 0.58?0.046 and 0.49?0.039,respectively,which were larger than that of MDA-MB-231BO(0.26?0.021,P
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Objective To explore the effects of fluvastatin on NF-?B activity and osteopontin(OPN) mRNA expression in albumin-induced renal tubular epithelial cells.Methods The renal tubular epithelial cells were cultured with 30 mg/ml fat free bovine serum albumin(BSA) as the control group.The renal tubular epithelial cells in the treatment group were cultured with different concentrations of fluvastatin for different hours.EMSA and RT-PCR were used to observe NF-?B activity and OPNmRNA expression.Results Fluvastatin can inhibit the NF-?B activity and OPNmRNA expression in a time and dose dependent manner.Conclusion Fluvastatin can inhibit NF-?B activity and OPNmRNA expression in albumin induced tubular epithelial cells.
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Sialic acid residues are constant constituents of the glycoproteins of the airways in all species. Sialoglycoproteins are the main acidic glycoprotein and their functions are to mediate cell adherence, to control the viscoelasticity of mucus and to serve as receptor sites for the binding of exogenous macromolecules. The purpose of this study was to investigate the differences in the distribution of sialoglycoproteins as a terminal sugar and in the composition of the penultimate sugar according to aging in the murine nasal respiratory and olfactory mucosa. Nasal cavities of mice (BALB/c) were fixed by intracardiac perfusion with 2.0% glutaraldehyde and embedded in Epon 812. First, the serial sections were stained with Maackia amurensis agglutinin (MAA) and Sambucus nigra agglutinin (SNA). Then, the adjacent sections were stained with DBA and PNA before and after neuraminidase digestion in all experimental groups. Apical cell surfaces of olfactory mucosa and cilia on a few ciliated cells in the mucosa of the septum and nasal floor were labelled with MAA, but cell surfaces of respiratory mucosa, Bowman's glands and goblet cells were not labelled with MAA, irrespective of aging. Apical cell surfaces of both olfactory and respiratory mucosa and Bowman's glands were stained with SNA, however, goblet cells were not labelled with SNA. After neuraminidase digestion to remove terminal sialic acid residues of sialoglycoproteins, only cell surfaces of respiratory mucosa were labelled with PNA, but goblet cells, cell surfaces of olfactory mucosa and Bowman's glands were not labelled with PNA. Cell surfaces and Bowman's glands of olfactory mucosa were labelled with DBA after neuraminidase digestion, but cell surfaces of respiratory mucosa and goblet cells were not labelled with DBA. Our results indicate that there were different carbohydrate structures of sialoglycoconjugates in olfactory and respiratory mucosa, and it was not influenced by aging.
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Mice , Aging/metabolism , Animals , Carbohydrates/analysis , Mice, Inbred BALB C , Nasal Mucosa/chemistry , Olfactory Mucosa/chemistry , Sialoglycoproteins/analysisABSTRACT
Objective:To express human dentin sialoprotein (hDSP) gene in COS-7 cells. Methods:hDSP gene was subcloned into mammalian expression vector pcDNA3. The recombined plasmids were transfected into COS-7 cells using lipofectamune PLUS TM kit for transient expression. Western blot analysis and immunohistochemical staining were used to examine the gene products. Results:The constructed vectors were confirmed by digestion with restriction enzyme. An immuno-reaction positive band with relative molecular mass of 60 000 was found by Western blot analysis in culture supernatant and cytoplasms of COS-7 cells. Immunohistochemical staining showed strong positive particles in the cytoplasms. Conclution:hDSP gene can be expressed in COS-7 cells.
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Objective Recent researches show that bone sialoprotein is closely related with metastasis of breast cancer to bone. In order to investigate the effect of bone sialoprotein in the course of metastasis of breast cancer to bone, the following study was undertaken. Methods The growth curve, clone formation and adherence of breast cancer cells experiments were carried out to study the effect of hBSP over-expression on biological behaviors of breast cancer cells. Results It was found that breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID) stably transfected with hBSP cDNA were not inhibited through proliferation assay of breast cancer cells. The activity and tumorigenicity of breast cancer cells with hBSP over-expression were enhanced. Comprising with MDA-MB-231BO non-transfected the colony formation percentage of breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID) increased from 8.90%?0.53% to 15.6%?0.94% and 12.8%?0.77%. The OD_ 620 value of adherence of breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID)increased from 0.29?0.017 to 0.52?0.031 and 0.43?0.026 respectively in cells attachment assay, which suggested that over-expression of hBSP enhances breast cancer cell MDA-MB-231BO adherence and attachment property. Conclusion The study results suggested that hBSP over-expression had autocrine effect on breast cancer cells and could enhance the ability of breast cancer cells metastasizing to bone.