Urine spermine and multivariable Spermine Risk Score predict high-grade prostate cancer.

BACKGROUND: To investigate the role of urine spermine and Spermine Risk Score in prediction of high-grade prostate cancer (HGPCa, ISUP grade group ≥2). METHODS: Nine hundred and five consecutive men with elevated PSA were prospectively recruited from two hospitals. Core analyses focused on consecutive men with PSA 4-20 ng/mL (n = 600). Pre-biopsy urine without prior prostatic massage was analyzed for spermine level with ultra-high performance liquid chromatography with triple quadrupole mass spectrometer (UPLC-MS/MS). The proportions of PCa and HGPCa were compared across different spermine ranges. Logistic regressions were used to form different models, and their performances were compared using area under curve (AUC) and decision curve analysis (DCA). RESULTS: PCa and HGPCa were diagnosed in 30.8% (185/600) and 17.2% (103/600) men, respectively, and were significantly associated with lower urine spermine levels. Between the lowest and highest quartiles of spermine results, a threefold increase in PCa risk (49.3% vs. 16.7%) and 3.5-fold increase in ISUP grade group ≥2 PCa risk (31.3% vs. 8.7%) were observed. Multivariate analysis showed PSA, prostate volume (PV), digital rectal examination (DRE), and spermine, which were independent predictors for PCa and HGPCa, and a Spermine Risk Score with these factors achieved the highest AUC of 0.78 for PCa and 0.82 for HGPCa. At 90% sensitivity for HGPCa, 36.7% biopsies and 24.4% ISUP grade group 1 diagnoses could have been avoided, with a negative predictive value of 95.4%. DCA revealed net clinical benefit of the Spermine Risk Score. Internal validation with bootstrapping showed good discrimination and calibration. CONCLUSION: Urine spermine and Spermine Risk Score identified men at higher risk of HGPCa and reduced unnecessary biopsies.

A novel homozygous splice site mutation in COL7A1 in a Chinese patient with severe recessive dystrophic epidermolysis bullosa and squamous cell carcinoma.

BACKGROUND: Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited blistering skin disorder caused by mutations in COL7A1 gene encoding type VII collagen, the major component of anchoring fibrils in the dermo-epidermal junction. The development of cutaneous squamous cell carcinoma (SCC) is one of the most serious complications of this disease. We report herein a Chinese patient with the severe generalized subtype of RDEB (RDEB-sev gen) complicated by SCC. METHODS: Skin biopsies were examined for histology, basement membrane ultrastructure, and type VII collagen expression. Genomic DNA was extracted from the peripheral blood samples and subjected to polymerase chain reaction amplification and direct automated DNA sequencing. RESULTS: Histopathological examination of the patient's skin revealed an undetectable expression of type VII collagen polypeptides in the basement membrane zone. Mutation analysis identified a novel splice site mutation in intron 64 (IVS64+5g->a) of COL7A1 gene, which resulted in an in-frame deletion of exon 64 in both alleles. CONCLUSIONS: This report contributes to the expanding database of COL7A1 mutations and emphasizes the need to elucidate the underlying genetic mechanisms associated with the increased incidence of SCC in RDEB patients.

Stem cell-like properties of human umbilical cord lining epithelial cells and the potential for epidermal reconstitution.

BACKGROUND AIMS: Stem cells are particularly attractive for many cell-based therapeutic interventions because of their ability to self-renew and their capacity to differentiate into site-specific differentiating cells. Restoration of the integrity of epithelial continuity is an essential aspect of wound repair and tissue regeneration. We are currently looking at the potential of human umbilical cord lining cells as a source of epithelial stem cells with appropriate differentiation capacity for potential epidermal reconstitution. METHODS: We isolated human umbilical cord lining epithelial cells (CLEC) and characterized their phenotype from the perspective of proliferative potential, telomere length, expression of epidermal differentiation markers, as well as stem cell-specific markers, and clonogenicity. Their potential for epidermal reconstitution was investigated in an organotypic culture model. RESULTS: The results demonstrated that CLEC present a long telomere length and have a relatively high proliferative potential and passaging ability in culture. CLEC display some of the stem cell-specific markers for epithelial as well as pluripotent stem cells, including CK19, p63, OCT-4, SSEA-4, TRA-1-60, SOX2 and Nanog. CLEC are capable of generating a fully stratified epithelium in organotypic culture. CONCLUSIONS: The potential of CLEC to be used in clinical applications for specialized epithelial reconstruction is still unexplored. The demonstration that CLEC have stem cell-like properties and are capable of generating fully stratified epithelium provides support for their potential clinical application in epidermal reconstitution.

HA modulation of epidermal morphogenesis in an organotypic keratinocyte-fibroblast co-culture model.

Hyaluronan (HA) is a major extracellular matrix component of the skin. Amongst its biological functions is the maintenance of epidermal homeostasis. The mechanisms of action, however, remain unclear. To explore the interaction of HA with the epidermis, we have looked at the effects of exogenous application of HA in an organotypic culture model containing a dermal substrate with and without fibroblast incorporation. The results demonstrate that exogenous HA enhances epidermal proliferation resulting in a thicker viable epidermis with an increase in the number and intensity of Ki67-positive basal cells; HA also improves the basement membrane assembly as evidenced by an increased expression of laminin-332 and collagen type IV but not the expression of Nidogen-1 at the epidermal-dermal junctional zone; furthermore, the development of epidermal lipid barrier structure was enhanced. These findings provide evidence to support the therapeutic use of exogenous HA for treating skin disorders with aberrant epidermal homeostasis.

Expression of transforming growth factor beta isoforms and their roles in tendon healing.

Transforming growth factor beta (TGF-beta) plays active roles in tendon healing. However, the differential effects of TGF-beta isoforms on tendon healing have not been investigated. In cultured tendon fibroblasts, we tested the effects of TGF-beta1, beta2, and beta3 on the mRNA levels of COL1A1 and COL3A1 by quantitative real-time polymerase chain reaction. We also investigated the expression of TGF-beta isoforms, TGF-beta receptors, procollagen Type I and Type III in a rat model of tendon healing. We found that TGF-beta3 exhibited the highest potency in stimulating COL1A1 and COL3A1. TGF-beta1 exerted antagonistic effects to TGF-beta2 and beta3. All TGF-beta isoforms and procollagen Type I were confined to the edges of the healing tendon at day 28 postinjury. Our results indicated that interaction of TGF-beta isoforms exist in the regulation of collagen synthesis in tendon fibroblasts. Their effects may be further complicated by uneven spatial distribution of TGF-beta and TGF-beta receptors in healing tendons.

TGF-beta1 reverses the effects of matrix anchorage on the gene expression of decorin and procollagen type I in tendon fibroblasts.

Transforming growth factor-beta1 is known for its effect on the production of extracellular matrix in tendons. Elevated levels of transforming growth factor-beta1 have been reported in tendon adhesion and tendinosis, which suggests that transforming growth factor-beta1 plays an important role in matrix disturbances. Tendon adhesion involves excessive collagen deposition, whereas tendinosis is associated with increased proteoglycan deposition. It seems that other factors also may affect matrix deposition and modulate the effects of transforming growth factor-beta1. We assessed whether matrix anchorage to Type I collagen or fibronectin could change the gene expression of matrix proteins in tendon fibroblasts, and studied whether the effects of transforming growth factor-beta1 were altered by matrix anchorage. Human patellar tendon fibroblast cultures were prepared in different cell anchorages, and the cellular responses to transforming growth factor-beta1 were measured as gene expression of procollagen Type I, Type III, decorin, and biglycan by real-time reverse transcriptase-polymerase chain reaction. Fibronectin anchorage significantly increased the messenger ribonucleic acid level of decorin, and the messenger ribonucleic acid level of procollagen Type I was decreased by matrix anchorage to either fibronectin or Type I collagen. Transforming growth factor-beta1 increased the messenger ribonucleic acid level of procollagen Type I in Type I collagen-coated plates, but it suppressed the messenger ribonucleic acid level of decorin in fibronectin-coated plates. These findings suggest that interaction of matrix anchorage and transforming growth factor-beta1 is an important determinant of matrix deposition in healing tendons and the development of matrix disturbances in tendons.

The roles of bone morphogenetic protein (BMP) 12 in stimulating the proliferation and matrix production of human patellar tendon fibroblasts.

Recombinant human (rh) bone morphogenetic protein 12 (BMP12) is proved to induce the formation of tendon and ligament tissues in animal experiments. But the roles of BMP12 on tissue regeneration in human tendons remain unexplored. In the present study, healthy human patellar tendon samples were collected for histological examination and preparation of tendon fibroblast culture. Immunohistochemical staining showed that BMP12 was detected on healthy patellar tendon samples, only located on active tenoblasts and perivascular mesenchymal cells but not in interstitial tenocytes. The expression of PCNA and procollagen type I also exhibited a similar distribution. It indicates that BMP12 may be involved in matrix remodeling process in adult tissues. In vitro studies showed that rhBMP12 could increase proliferation of tendon fibroblasts and increase the gene expression of procollagen type I and type III, but decrease the gene expression of decorin in tendon fibroblasts culture. Our findings suggest that BMP12 may play a role in early phases of tissue regeneration in tendons.

Increased expression of transforming growth factor-beta1 in patellar tendinosis.

Patellar tendinosis is characterized by longstanding localized and activity-related pain, swelling and tenderness on palpation, and characteristic features on magnetic resonance imaging and ultrasonography and during surgical excision. Histologic examination of tendinosis tissues shows disrupted collagen matrix, increased cellularity, and increased proteoglycan stainability, but lack of inflammatory cell infiltration despite the clinical signs resembling inflammation. Disturbances in inflammatory response may be associated with the development of tendinosis. Expression of cyclooxygenase-2 and transforming growth factor-beta1 were detected in tendinosis, and the in vitro production of prostaglandin E2 by tendinosis and healthy tendon fibroblast cultures also was observed. Eleven patients with patellar tendinosis and 12 control subjects with healthy patellar tendons, but deficient anterior cruciate ligaments, were included in the current study. The percentages of immunopositive cells in tendinosis samples for cyclooxygenase-2 and transforming growth factor-beta1 were 66.75 and 56.40, respectively, which were significantly higher than those of the control subjects (25.15 and 23.06 respectively). Tendinosis fibroblast culture also produced more prostaglandin E2 and active transforming growth factor-beta1. These findings indicate the involvement of prostaglandins and cytokines that may explain the clinical symptoms and nonhealing features of tendinosis.