Advanced atherosclerosis is often associated with dystrophic calcification and remodeling of extracellular matrix of vascular wall. Recently many studies have documented a general relationship between calcification and severity of coronary disease, and discussed the feasibility of electron beam computed tomography for detecting and quantifying the coronary artery calcification in the patients. The present study investigated the expression and the localization of osteopontin, one of noncollagenous bone matrix protein, within the calcified coronary arteries. Autopsy-derived coronary artery specimens were scanned and reconstructed to visualize the pattern of coronary calcification using a novel microscopic computed tomography technique. The localization of the osteopontin were evaluated by immunohistochemial stain with LF7. The present study showed that the pattern of coronary calcification is variable and the expression of osteopontin is localized mainly to calcified lesion. The smooth muscle cells in addition to macrophage expressed osteopontin protein in human coronary atherosclerotic plaques. Soluble osteopontin released near to the sites of vascular calcification may represent an adaptive mechanism aimed at regulating the process of vascular calcification.
Aged ; Calcinosis/metabolism ; Coronary Arteriosclerosis/pathology* ; Coronary Arteriosclerosis/metabolism* ; Coronary Vessels/pathology* ; Coronary Vessels/metabolism ; Coronary Vessels/chemistry* ; Female ; Human ; Immunohistochemistry ; Male ; Middle Age ; Sialoglycoproteins/biosynthesis ; Sialoglycoproteins/analysis*

Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood except the suppression of osteoblast function. To investigate the effect of simulated weightlessness on mRNA level of extracellular matrix proteins, osteoblasts were rotated in horizontal plane as a model of simulated microgravity. Primordial osteoblasts of rats were grown for 2 d and then rotated for 24, 48 and 72 h, respectively. After isolating total RNA in cells, reverse transcription polymerase chain reaction (PT-PCR) was made to examine the mRNA level of osteopontin (OPN) and osteonectin (ON). Meanwhile, the levels of alkaline phosphatase (ALP) and osteocalcin (BGP) in the cultured medium were measured to evaluate the calcific function of cell. The expression of OPN and ON mRNA fell significantly after rotating for 24, 48 and 72 h, respectively. The contents of BGP descended significantly, meanwhile, the activity of ALP also showed a degressive tendency. Horizontal rotation decreased the expression of ON and OPN as well as diminished the secretion of BGP and ALP, which affected the calcific function of osteoblast. The results obtained suggest that depression of extracellular matrix proteins expression plays a key role in bone loss during weightlessness.
Animals ; Animals, Newborn ; Cells, Cultured ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteonectin ; biosynthesis ; genetics ; Osteopontin ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Rotation ; Sialoglycoproteins ; biosynthesis ; genetics ; Skull ; cytology ; Weightlessness Simulation

OBJECTIVETo observe the effect of Astragalus-Angelica Mixture (AAM) on osteopontin (OPN) expression in rats with chronic nephrosclerosis.

METHODSChronic nephrosclerosis model rats induced by repeated intraperitoneal injection of puromycin were randomly divided into the model group, AAM group and Irbesartan (an antagonist of angiotensin) group. The experimental course lasted 12 weeks. Blood and urine samples were examined by biochemical method. Kidney tissue was taken for pathological stain and immunohistochemical method and was applied to examine OPN expression, mononuclear macrophage, laminin in extracellular matrix and decorin expressions.

RESULTSAAM showed the effects of decreasing urinary protein and improving renal function similar to that of Irbesartan. It also could alleviate the pathological damage of kidney tissue, especially in decreasing renal tubular mesenchymal damage index. The accumulation of decorin and laminin in the mesenchymal extracellular matrix significantly decreased. Renal tubular OPN expression and mesenchymal infiltration of mononuclear macrophage decreased significantly and in a positive correlated manner (r = 0.885, P < 0.01).

CONCLUSIONAAM has similar renal protective action to that of Irbesartan, this action may be related to the inhibition of up-regulated OPN expression.

Animals ; Astragalus Plant ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Nephrosclerosis ; chemically induced ; metabolism ; Osteopontin ; Phytotherapy ; Puromycin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; biosynthesis

To estimate intracellular interleukin-1 receptor antagonist (icIL-1ra) expression in the bone marrow cells from adult patients with chronic myelogenous leukemia (CML), flow cytometry (FCM) assay was used for detecting the mean icIL-1ra fluorescence intensity per cell (equivalent to it's expression level) in different groups of cells from normal and CML patient bone marrows by 15 monoclonal antibodies with different fluorescent marker. The results showed that all of marrow nucleated cells express IL-1ra, but its expression level in granulocytes was the highest and that in lymphocytes was the lowest. The icIL-1ra expression level was significantly lower in nucleated marrow cells, granulocytes and lymphocytes from the marrow of 17 untreated CML patients than that from the marrow of 8 normal. The mean icIL-1ra fluorescence intensity was significantly lower in marrow nucleated cells, granulocytes and lymphocytes in 13 CML patients with marrow blasts >or= 10% than that in 43 CML patients with marrow blasts < 5% or than that in 9 CML patients with marrow blasts 5% - 10%. It was concluded that the lower expression of icIL-1ra in CML marrow nucleated cells might be involved in the evolution of CML.
Adult ; Bone Marrow Cells ; metabolism ; Flow Cytometry ; Humans ; Interleukin 1 Receptor Antagonist Protein ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Sialoglycoproteins ; biosynthesis ; blood

OBJECTIVETo investigate the effects of specific RNA interference (RNAi) to Notch ligand Delta1 on proliferation and differentiation of human dental pulp stem cells (DPSC).

METHODSDPSC were infected by lentivirus vectors carrying Delta1-RNAi. DPSC were divided into three groups, DPSC/Delta1-RNAi group, DPSC/wt group and DPSC/vector group as control. Cell counting kit-8 (CCK-8) assay and flow cytometry were used to evaluate the proliferation of DPSC. Expression of proliferating cell nuclear antigen (PCNA) was examined with immunohistochemical staining. All groups were cultured in an odonto-inductive medium and were observed under microscope. The number of mineralization nodules was counted after Alizarin red staining. Alkaline phosphatase (ALP) activity and the expression of dentin sialophosphoprotein (DSPP) were detected by ALP activity assay and Western blotting.

RESULTSCompared with DPSC/wt or DPSC/vector separately, proliferating rate and S-cycle of DPSC/Delta1-RNAi was significantly lower. The S phase and proliferation index (PI) decreased markedly from 22.32 ± 2.35 and 33.68 ± 4.19 (DPSC/Delta1-RNAi) to 5.44 ± 0.91 and 16.00 ± 6.07 (DPSC/wt). The PCNA staining of DPSC/Delta1-RNAi was evidently weaker. DPSC/Delta1-RNAi group had more calcified cell nodules than the other two control groups, and ALP activity and DSPP expression of DPSC/Delta1-RNAi group increased markedly.

CONCLUSIONSDelta1-RNAi induced by the lentivirus vectors may inhibit DPSC proliferation and differentiation. Notch-Delta signal pathway plays an important role in self-renewal and differentiation.

Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Dental Pulp ; cytology ; metabolism ; Epithelial Cells ; Extracellular Matrix Proteins ; biosynthesis ; Genetic Vectors ; Homeodomain Proteins ; Humans ; Lentivirus ; Phosphoproteins ; biosynthesis ; RNA Interference ; Sialoglycoproteins ; biosynthesis ; Signal Transduction ; Stem Cells ; metabolism

The aims of this study were to investigate the relationships between the production of interleukin-1 (IL-1), and IL-6 system by whole blood cells, and bone mineral density (BMD), and polymorphisms in IL-1 system and IL-6 gene in postmenopausal Korean women. The production of IL-1alpha, IL-1beta, IL-1 receptor antagonist (IL-1ra), IL-6, and soluble IL-6 receptor (sIL-6r) by lipopolysaccharide-stimulated whole blood cells was measured by ELISA in 110 subjects. Serum osteocalcin, C-telopeptide of type I collagen, and BMD at lumbar spine and proximal femur were measured. IL-1alphaC(-889)T polymorphism, IL-1beta C(-511)T polymorphism, 86-base pair variable number tandem repeat polymorphism in the IL-1ra gene, and IL-6 C(-634)G polymorphism were analyzed. The production of IL-1beta correlated positively with BMD at femoral neck, whereas the production of other ILs did not correlate with BMD at the skeletal sites examined. No significant differences in the production of ILs were observed among normal, osteopenic and osteoporotic postmenopausal women, and among the different IL system polymorphisms groups studied. No correlation between bone turnover markers and the production of ILs was noted. In conclusion IL-1beta may regulate bone metabolism at femoral neck, and the IL system polymorphism do not affect the production of ILs by whole blood cells.
Aged ; Blood Cells/drug effects/immunology ; Bone Density/*genetics/*immunology ; Bone Diseases, Metabolic/blood/genetics/immunology ; Cytokines/*biosynthesis/blood ; Female ; Humans ; In Vitro ; Interleukin-1/biosynthesis/blood/genetics ; Interleukin-6/biosynthesis/blood/genetics ; Interleukins/*genetics ; Lipopolysaccharides/pharmacology ; Middle Aged ; Osteoporosis, Postmenopausal/blood/genetics/immunology ; *Polymorphism, Genetic ; Receptors, Interleukin-6/biosynthesis/blood/genetics ; Research Support, Non-U.S. Gov't ; Sialoglycoproteins/biosynthesis/blood/genetics

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adenocarcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Organic Cation Transport Proteins ; biosynthesis ; genetics ; Organic Cation Transporter 2 ; Osteopontin ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Sialoglycoproteins ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate