OBJECTIVETo observe the histological characteristics of constructed basement membrane in tissue-engineered skin.

METHODSForskins from circumcision in normal children were obtained with informed consent of the parents, and then the epidermal keratinocytes (KC) and dermal fibroblasts (Fb) were isolated with trypsin and collagenase D digestion in sequence. Tissue engineered skin with composite chitosan was maintained in a submerged state for 3 days, and then at the air-liquid interface. The tissue-engineered skins were fixed in neutral formalin and then embedded in paraffin after culture for 7, 10 and 15 days, respectively for immunohistological examination of the basement membrane component,including the condition of collagen type IV (COL-IV), collagen type VII (COL-VII), and laminin (LN).

RESULTSHE staining showed that the keratinocytes formed a fine stratified squamous epithelium with the presence of basal, spinous, granular and corneous cell layers, and there was various amount of cells in flat and fusiform shape in each layer. It was found that a regular red staining strip situated at the dermal epidermal junction. Positive staining of collagen IV, collagen VII as well as LN was observed by immunohistological examination.

CONCLUSIONThe results suggest that the composite chitosan tissue engineered skin has a good prospect for clinical use because it presents a perfect reconstruction of basement membrane.

Basement Membrane ; cytology ; Cells, Cultured ; Child ; Chitosan ; metabolism ; Collagen Type IV ; metabolism ; Collagen Type VII ; metabolism ; Humans ; Laminin ; metabolism ; Organ Culture Techniques ; Skin, Artificial ; Tissue Engineering ; methods

Non-enzymatic nitrite induced collagen cross-linking results in changes reminiscent of age-related damage and parallels the well-known model system, non-enzymatic glycation. We have recently observed that nitrite modification of basement membrane proteins can induce deleterious effects on overlying retinal pigment epithelial cells in studies relevant to age-related macular degeneration. The present work was undertaken in order to confirm 3-nitro-tyrosine (3-NT) as a product of the reaction and to identify the site specificity of nitration in collagen IV, a major component of basement membranes. Human collagen type IV was modified via incubation with 200 mM NaNO2 (pH=7.38) for one week at 37degrees C. The modified protein was prepared in 2 different ways, including acid hydrolysis and trypsin digestion for site specificity determination. The samples were analyzed by LC/MS using a C12 RP column. Site specificity was determined from tandem MS/MS data utilizing TurboSEQUEST software and the Swiss-Prot sequence database. 3-NT was detected in protein digests and acid hydrolysates of nitrite modified collagen IV. Positive identification with standard 3-NT was confirmed by identical Rt, lambda(max)=279 nm and 355 nm, and m/z=227. Analyses of tryptic digests identified four sites of tyrosine nitration, alpha1(IV)Y348, alpha1(IV)Y534, alpha2(IV)Y327, and alpha2(IV)Y1081. These sites are located in the triple-helical region of the protein and provide clues regarding potential sites for nitrite modification in collagen type IV.
Tyrosine/*metabolism ; Tandem Mass Spectrometry ; Substrate Specificity ; Nitrites/*metabolism ; Humans ; Collagen Type IV/*metabolism ; Chromatography, Liquid ; Binding Sites

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

Actins ; metabolism ; Animals ; Collagen Type III ; metabolism ; Collagen Type IV ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; metabolism ; pathology ; HSP47 Heat-Shock Proteins ; metabolism ; Kidney Glomerulus ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vimentin ; metabolism

OBJECTIVETo explore the relationship between the extracellular matrix and apoptosis of intestinal epithelium after burn injury.

METHODSThirty Wistar rats were employed in the study and were randomly divided into normal control (C) and 6 PBH (postburn hour), 12 PBH, 1 PBD (postburn day), 3 PBD and 5 PBD group with 5 rats in each group. The rats in burn groups were sacrificed at 0, 6 and 12 PBHs and 1, 3 and 5 PBDs. The apoptotic cell count and the caspases-3 activity of intestinal mucosal epithelium, and the extracellular matrix component laminin and type IV collagen were determined and their correlation was analyzed.

RESULTSThe apoptotic cell count and the caspases-3 activity of intestinal mucosal epithelium in burn groups were obviously higher than those in C group (P < 0.05 or 0.01), while the intestinal mucosal contents of laminin and type IV collagen in burned groups were much lower than those in C group (P < 0.05 or 0.01). By linear correlative analysis, it was shown that the changes in the intestinal mucosal contents of laminin and type IV collagen in burned groups were negatively correlated with the change in apoptotic cell count (r = -0.575, -0.613, P < 0.05).

CONCLUSIONIntestinal epithelial apoptosis was enhanced after burn injury, and it was correlated with the change in the components of the extracellular matrix.

Animals ; Apoptosis ; Burns ; metabolism ; pathology ; Caspase 3 ; metabolism ; Collagen Type IV ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Intestinal Mucosa ; metabolism ; pathology ; Laminin ; metabolism ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study the expression and significance of structural proteins, VEGF and Ang-1 in cavernous venous malformations of the body surface.

METHODSTissue samples came from 25 cases of cavernous venous malformations, 12 cases of normal moderate veins and 12 cases of normal small veins. Envision immunohistochemical stain was used to investigate the expression of IV collagen, fibronectin, laminin, VEGF and Ang-1. The results were analyzed semi-quantitatively.

RESULTSThe distribution of structural proteins in cavernous venous malformations is similar to moderate and small veins, but the expression in venous malformations is less obviously. VEGF expression in cavernous venous malformations and small veins is stronger obviously than moderate veins. Ang-1 expression in small veins is stronger remarkably than cavernous venous malformations and moderate veins.

CONCLUSIONThe abnormal expression of structural proteins may be an important factor in etiopathology and progress of cavernous venous malformations. There is disturbance of blood vessel remodelling in the sinusoid of cavernous venous malformations, with which the less expression of Ang-1 may be related.

Angiopoietin-1 ; metabolism ; Collagen Type IV ; metabolism ; Fibronectins ; analysis ; Hemangioma, Cavernous ; metabolism ; Humans ; Laminin ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Veins ; abnormalities ; metabolism

Skin wound healing is a complex event, and interrupted wound healing process could lead to scar formation. The aim of this study was to examine the morphological changes of scar tissue. Pathological staining (HE staining, Masson's trichrome staining, methenamine silver staining) was used to evaluate the morphological changes of regenerating epidermis in normal skin and scar tissue, and immunofluorescence staining to detect the expression of collagen IV, a component of basement membrane (BM), and the expression of integrinβ4, a receptor for BM laminins. Additionally, the expression of CK14, CK5, and CK10 was measured to evaluate the proliferation and differentiation of keratinocytes in normal skin and scar tissue. The results showed that the structure of the skin was histologically changed in scar tissue. Collagen IV, expressed under the epidermis of normal skin, was reduced distinctly in scar tissue. Integrinβ4, expressed in the basal layer of normal skin, was found absent in the basal layer of scar tissue. Additionally, it was found that keratinocytes in scarring epidermis were more proliferative than in normal skin. These results indicate that during the skin wound healing, altered formation of BM may affect the proliferation of keratinocytes, reepithelial and tissue remodeling, and then result in scar formation. Thus, remodeling BM structure during wound repair may be beneficial for improving healing in cutaneous wounds during clinical practice.
Adolescent ; Adult ; Cicatrix ; metabolism ; pathology ; Collagen Type IV ; metabolism ; Female ; Humans ; Integrin beta4 ; metabolism ; Keratinocytes ; cytology ; metabolism ; pathology ; Male ; Skin ; cytology ; metabolism ; pathology

OBJECTIVEIn order to investigate whether or not thrombopoietin (TPO) could promote the fibrogenesis of bone marrow stromal cells in absence of megakaryocytes (MKs).

METHODSImproved dexter culture system with various TPO concentrations was used for ex vivo culture of bone marrow stromal cells. Relative proliferation index, the expressions of fibronectin, laminin and type IV collagen, and the systhesis of type III procollagen were detected at different time points during culture process.

RESULTSTPO stimulated the proliferation of bone marrow stromal cells. Relative proliferation index of the stromal cells increased with the TPO concentration increasing, and was not related to the exposure time. The expressions of fibronectin, laminin, and type IV collagen appeared stronger in the TPO groups than those in the control group. But the expressions of these molecules were not dependent upon the culture time. TPO could accelerate the synthesis of type III procollagen in bone marrow stromal cells, and this acceleration was unrelated to the TPO concentration.

CONCLUSIONThese findings suggested that TPO could stimulate the stromal cells with a consequence of increased syntheses and secretions of the extracellular matrix and collagen in absence of MKs. In other words, TPO could promote the fibrogenesis of bone marrow stromal cells without the existence of MKs.

Cells, Cultured ; Collagen Type III ; metabolism ; Collagen Type IV ; metabolism ; Extracellular Matrix ; metabolism ; Fibronectins ; metabolism ; Fibrosis ; pathology ; Humans ; Laminin ; metabolism ; Megakaryocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; pathology ; Thrombopoietin ; pharmacology

OBJECTIVE: To investigate the expression of collagen type I (Col I), collagen type IV (Col IV) and fibronectin (Fn) in laryngeal squamous cell carcinomas (LSCC) and their pathobiological relationship with invasion and metastasis of tumor. METHOD: The expression of Col I, Col IV and Fn was detected by immunohistochemistry method in normal tissue of latero-carcinoma and tissue of carcinoma in 60 specimens of LSCC. Integral optical density (iOD) was detected by image analysis of computer and was analyzed by SPSS13.0. RESULT: The expression of Col I was obvious and integrity. The expression of Col IV and Fn of basal membrane was like intact line-shape appearance and Fn of interstitial substance appeared like a complete network in the normal tissue of latero-carcinoma. Their expression decreased gradually and their integrity was broken and disappeared gradually from well to poorly differentiated LSCC. Their expressions also fell off with the tumor size, clinical stage and cervical lymphatic metastasis gradually and consistently in LSCC. Their variances were statistical significance separately (P<0.05 or P<0.01). CONCLUSION The expression of Col I, Col IV and Fn was closely related to tumor invasion, the regional lymph node metastasis and other some pathobiological features in LSCC. A detection of Col I, Col IV and Fn was of definite significance on a better comprehension of the possibility of metastasis, choice of surgery and prognosis.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Collagen Type I ; metabolism ; Collagen Type IV ; metabolism ; Female ; Fibronectins ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Larynx ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis

Breast Neoplasms ; pathology ; Carcinoma in Situ ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Carcinoma, Papillary ; metabolism ; pathology ; Collagen Type IV ; metabolism ; Female ; Humans ; Neoplasm Invasiveness ; pathology