OBJECTIVETo investigate the correlation and anatomic association of benign hyperplastic nodules in the peripheral zone (PZ) with those in the transition zone (TZ) of the prostate, and to compare the histological components of the two kinds of nodules.

METHODSWe obtained benign hyperplastic nodules specimens from the PZ and TZ by autopsy, measured the distance between the outer surface of the nodules and the inner gland, observed the integrity of the surgical envelope of the prostate, and determined the histological components of the two kinds of nodules by HE staining, immunohistochemistry and automatic quantitative image analysis.

RESULTSThe surgical envelope of the prostate was integrated and the distance between the nodules of the PZ and the outer surface of the inner gland was about 2.5 to 5 mm ([3.9 +/- 0.8] mm), with no signs of anatomic connection in between. The stromata and epithelia in the nodules accounted for (69.32 +/- 8.35)% and (16.08 +/- 5.36)% in the PZ and (74.58 +/- 8.95)% and (15.82 +/- 6.41)% in the TZ.

CONCLUSIONBenign hyperplastic nodules may originate from the PZ of the prostate and not correlate with the inner gland hyperplasia in the TZ, but with no statistical difference between the histological components of the two kinds of nodules.

Aged ; Aged, 80 and over ; Autopsy ; Collagen Type I ; analysis ; Collagen Type II ; analysis ; Collagen Type III ; analysis ; Collagen Type IV ; analysis ; Fibronectins ; analysis ; Humans ; Hyperplasia ; Immunohistochemistry ; Laminin ; analysis ; Male ; Prostate ; chemistry ; pathology ; Prostatic Hyperplasia ; metabolism ; pathology

BACKGROUND: Changes in the composition of the extracellular matrix (ECM) occur between the proliferating and involuted phases of infantile hemangiomas (IH), and are associated with angiogenic growth. We examined the composition of the ECM in proliferating and involuted IHs and assessed correlations between the composition of the ECM and whether the IH was in the proliferating or the involuted phase. METHODS: We evaluated IH samples from a cohort of patients who had five proliferating IHs and five involuted IHs. The following ECM molecules were analyzed using enzyme-linked immunosorbent assays and immunohistochemistry: laminin, fibronectin, collagen type I, collagen type II, and collagen type III. RESULTS: The involuted IHs had higher levels of deposition of collagen type III than the proliferating IHs. The median values (interquartile ranges) were 1.135 (0.946-1.486) and 1.008 (0.780-1.166) (P=0.019), respectively. The level of laminin was higher in involuted IHs than in proliferating IHs, with median values (interquartile ranges) of 3.191 (2.945-3.191) and 2.479 (1.699-3.284) (P=0.047), respectively. Abundant collagen type III staining was found in involuted IHs. Laminin alpha4 chain staining was clearly present within the basement membrane adjacent to the blood vessels, and was significantly more intense in involuted IHs than in proliferative IHs. CONCLUSIONS: Involuted hemangiomas showed extensive deposition of collagen III and laminin, suggesting that differences in the composition of the ECM reflect stages of the development of IHs. This pattern may be due to the rapid senescence of IHs.
Aging ; Basement Membrane ; Blood Vessels ; Cohort Studies ; Collagen ; Collagen Type I ; Collagen Type II ; Collagen Type III ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix* ; Fibronectins ; Hemangioma* ; Humans ; Immunohistochemistry ; Laminin

BACKGROUND: The fibrous proteins of extracellular matrix (ECM) produced by dermal fibroblast contributes to the maintenance of connective tissue integrity. OBJECTIVE: This study is carried out to identify the bioactive ingredient from natural products that enhances ECM production in dermal fibroblasts. METHODS: Bioassay-directed fractionation was used to isolate the active ingredient from natural extracts. The effects of rasatiol (isolated from Raphanus sativus) on ECM production in primary cultured human dermal fibroblasts was investigated by enzyme linked immunosorbent assay and western blot analysis. RESULTS: Rasatiol accelerated fibroblast growth in a dose-dependent manner and increased the production of type 1 collagen, fibronectin and elastin. Phosphorylation of p42/44 extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and Akt was remarkably increased by rasatiol, indicating that enhanced ECM production is linked to the activation of intracellular signaling cascades. CONCLUSION: These results indicate that rasatiol stimulates the fibrous components of ECM production, and may be applied to the maintenance of skin texture.
Biological Agents ; Blotting, Western ; Collagen Type I ; Connective Tissue ; Elastin ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Raphanus ; Scleroproteins ; Skin

BACKGROUND: The fibrous proteins of extracellular matrix (ECM) produced by dermal fibroblast contributes to the maintenance of connective tissue integrity. OBJECTIVE: This study is carried out to identify the bioactive ingredient from natural products that enhances ECM production in dermal fibroblasts. METHODS: Bioassay-directed fractionation was used to isolate the active ingredient from natural extracts. The effects of rasatiol (isolated from Raphanus sativus) on ECM production in primary cultured human dermal fibroblasts was investigated by enzyme linked immunosorbent assay and western blot analysis. RESULTS: Rasatiol accelerated fibroblast growth in a dose-dependent manner and increased the production of type 1 collagen, fibronectin and elastin. Phosphorylation of p42/44 extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and Akt was remarkably increased by rasatiol, indicating that enhanced ECM production is linked to the activation of intracellular signaling cascades. CONCLUSION: These results indicate that rasatiol stimulates the fibrous components of ECM production, and may be applied to the maintenance of skin texture.
Biological Agents ; Blotting, Western ; Collagen Type I ; Connective Tissue ; Elastin ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Raphanus ; Scleroproteins ; Skin

The preeclamptic nephropathy is characterized by swelling of endothelial cells, interposition of mesangial cells and matrix, subendothelial deposits of incompletely defined material, and thickening of the capillary walls. To determine the distribution of extracellular matrix (ECM) components in preeclamptic nephropathy, the immunohistochemical study was performed in ten renal biopsy cases using antisera to human type I, III, IV, and VI collagens, fibronectin, and laminin. In preeclamptic nephropathy, the accumulation of type IV and VI collagens, fibronectin was observed in moderate amount in the mesangium and, to some extent, in the thickened capillary walls, particularly in the subendothelial layer. In segmentally sclerotic lesions seen in six cases, the amount of type IV collagen was partly decreased, whereas those of type VI collagen and fibronectin were slightly increased. Type I collagen was expressed to a mild degree in the expanded mesangium and segmentally sclerotic lesions. The results suggest that the expression of ECM in the mesangium is increased in preeclamptic nephropathy, and the deposition of ECM components may be involved in the development and the reparative process of the characteristic glomerular lesions. The formation of sclerotic lesions may be linked to the alternative accumulation of ECM components.
Biopsy ; Capillaries ; Collagen ; Collagen Type I ; Collagen Type IV ; Collagen Type VI ; Endothelial Cells ; Extracellular Matrix* ; Fibronectins ; Humans ; Immune Sera ; Laminin ; Mesangial Cells

We performed photorefractive keratectomy(PRK) on 10 rabbit eyes and determined the distribution of collagen type III, IV VI, VII at postoperative 2, 4 and 6 months to examine immunohistochemical changes after PRK. Type III collagen was not found in the normal cornea but strongly detected in the regenerated corneal stroma at all intervals. It was most prominent at 2 months after surgery and then decreased. Type IV collagen was detected in basement membrane in both normal and ablated corneas at all intervals and the staining was more intense in ablatd corneas than in normal cornea. There was no difference of staining intensity among the groups of different intervals. Type IV collagen was found in both normal and healed corneal stroma at all intervals and there was no difference of staining intensity between normal and ablated corneas and among the groups of different intervals. Type VII collagen was observed as a linear continuous band along the basal surface of epithelium in normal cornea. At 2 months after surgery, type VII collagen staining in basement membrane zone became denser than normal cornea, but segmented. At 4 months after surgery, continuous band of collagen type VII staining was observed, but it was less intense than in normal cornea. At 6 months after surgery, the intensity of continuous band of collagen type VII was the same as in normal cornea. This results suggest that the presence of type III collagen in the regenerated cornea may be related to the development of postoperative subepithelial opacity after PRK and the normalization of collagen type IV and VII at postoperative 6 months may mean the complete reestablished of the adhesion of regenerated epithelium and stroma.
Basement Membrane ; Collagen Type III* ; Collagen Type IV ; Collagen Type VII ; Collagen* ; Cornea* ; Corneal Stroma ; Epithelium ; Immunohistochemistry ; Lasers, Excimer* ; Photorefractive Keratectomy

Keloids are characterized by excessive extracelluar matrix (ECM) deposition such as collagen, fibronectin, elastin, and proteoglycans in the dermis. Recently, the use of botulinum toxin A (BTXA) in the treatment of keloids have had good results. To investigate the therapeutic effect of BTXA on the keloids, we evaluated the mRNA expression of collagen type I, type III, MMP (matrix metalloproteinases)-1, and TIMP (tissue inhibitor of metalloproteinases)-1 on keloid fibroblasts (KFs, n=5) after administration of BTXA. We also evaluated the enzymatic activity of MMP-2 and 9 by using zymography with BTXA. The same process was repeated after administration of TGF-beta in addition to BTXA. Type III collagen mRNA expression was decreased significantly when BTXA was administrated on KFs regardless of the presence or absence of TGF-beta. MMP-1 mRNA expression in KFs was increased according to the BTXA concentration increment, however, not increased with TGF-beta. Moreover, MMP-2 enzymatic activity in KFs was increased when BTXA administrated regardless of the presence or absence of TGF-beta. These results suggest that the down regulation of collagen III expression, the up regulation of MMP-1, and increased MMP-2 enzymatic activity on KFs after BTXA administration are able to decrease the excess collagen deposition in keloids.
Botulinum Toxins ; Collagen ; Collagen Type I ; Collagen Type III ; Dermis ; Down-Regulation ; Elastin ; Fibroblasts ; Fibronectins ; Keloid ; Matrix Metalloproteinases ; Proteoglycans ; RNA, Messenger ; Tissue Inhibitor of Metalloproteinases ; Transforming Growth Factor beta ; Up-Regulation

This study was focused on the localizations or expressions of fibronectin and interactions of fibronectin with type I and type III collagen in the new forming tissues during wound healing in rat skin. Adult male rats(SPrague-Dawley strain), 150-200 gm weight, were used for experimental animals. And the experimental animals were divided into 3 or 4 groups by passed days after incision of back skin.The specimens were obtained and immunohistologically stained for electron microscopy from each of groups and the specimens were observed with the electron microscope (Hitach-600 Model). The results obtained were as follows. 1. Strong positive reactions for type I collagen are seen in the new forming tissue at day 3 after incision of skin, and moderate reactions at day 5 and day 7 after incision of skin. 2. The weak reactions for type III collagen are seen at day 3 and day 5 after incision in the new forming tissues of skin. And the type III collagen reaction at day 7 are more increased than that of other groups. 3. Fibronectin expressions are seen moderately at day 5 and day 3 groups, and strong at day 5 group, but at day 7 group reactions are rapidly decreased. It is suggested that type I and III collagens are gradually increased from the beginning of wound healing to the time of the new fibrous tissues formation. The great activities of the fibronection are seen within processes of the new tissue formation for wound healing events.
Adult ; Animals ; Collagen ; Collagen Type I ; Collagen Type III ; Fibronectins ; Humans ; Male ; Microscopy, Electron ; Rats ; Skin ; Wound Healing ; Wounds and Injuries

Both mature and stem cell-derived hepatocytes lost their phenotype and functionality under conventional culture conditions. However, the 3D scaffolds containing the main extracellular matrix constitutions, such as heparin, may provide appropriate microenvironment for hepatocytes to be functional. The current study aimed to investigate the efficacy of the differentiation capability of hepatocytes derived from human Wharton's jelly mesenchymal stem cells (WJ-MSCs) in 3D heparinized scaffold. In this case, the human WJ-MSCs were cultured on the heparinized and non-heparinized 2D collagen gels or within 3D scaffolds in the presence of hepatogenic medium. Immunostaining was performed for anti-alpha fetoprotein, cytokeratin-18 and -19 antibodies. RT-PCR was performed for detection of hepatic nuclear factor-4 (HNF-4), albumin, cytokeratin-18 and -19, glucose-6-phosphatase (G6P), c-met and Cyp2B. The results indicated that hepatogenic media induced the cells to express early liver-specific markers including HNF4, albumin, cytokeratin-18 and 19 in all conditions. The cells cultured on both heparinized culture conditions expressed late liver-specific markers such as G6P and Cyp2B as well. Besides, the hepatocytes differentiated in 3D heparinized scaffolds stored more glycogen that indicated they were more functional. Non-heparinized 2D gel was the superior condition for cholangiocyte differentiation as indicated by higher levels of cytokeratin 19 expression. In conclusion, the heparinized 3D scaffolds provided a microenvironment to mimic Disse space. Therefore, 3D heparinized collagen scaffold can be suggested as a good vehicle for hepatocyte differentiation.
Antibodies ; Collagen ; Collagen Type I ; Constitution and Bylaws ; Extracellular Matrix ; Fetal Proteins ; Gels ; Glucose-6-Phosphatase ; Glycogen ; Heparin ; Hepatocytes* ; Humans ; Keratin-18 ; Keratin-19 ; Mesenchymal Stromal Cells* ; Phenotype ; Wharton Jelly

To evaluate the clinical significance of autoantibodies to three major epithelial cytokeratins (CK) -- CK8, CK18, and CK19 -- we compared 66 patients with toluene diisocyanate (TDI)-induced asthma (group I) with three control groups: 169 asymptomatic exposed subjects (group II), 64 patients with allergic asthma (group III), and 123 unexposed healthy subjects (group IV). Serum IgG, specific for human recombinant CKs, were measured by ELISA (enzyme linked immunosorbent assay), and ELISA inhibition tests were performed. The existence of these antibodies was confirmed by IgG immunoblot analysis. Anti-TDI-HSA (human serum albumin) IgE and IgG antibodies were measured by ELISA in the same set of the patients. The prevalence of CK8, CK18, and CK19 auotantibodies in group I was significantly higher than in the other three groups. Results of the ELISA inhibition test showed significant inhibition with the addition of three CKs in a dose-dependent manner. No significant association was found between CK autoantibodies and the prevalence of anti- TDI-HSA IgG and IgE antibodies. These results suggest that autoantibodies to CK18 and CK19 can be used as serologic markers for identifying patients with TDI-induced asthma among exposed workers.
Toluene 2,4-Diisocyanate/*toxicity ; Sensitivity and Specificity ; Occupational Diseases/chemically induced/*diagnosis ; Middle Aged ; Male ; Keratins/*immunology ; Keratin-8/immunology ; Keratin-19/immunology ; Keratin-18/immunology ; Immunoblotting ; Humans ; Female ; Enzyme-Linked Immunosorbent Assay ; Biological Markers/blood ; Autoantibodies/*blood ; Asthma/chemically induced/*diagnosis ; Adult