Temporal expression of chitinase-like 3 in wounded murine skin.

In forensic practice, it is important to diagnose wound age accurately. We analyzed the proteome of injured murine skin to identify a novel protein marker of wound age after recent injury. We used samples from 3 days after injury, with 0 days as the control. The proteins were separated with two-dimensional electrophoresis. Using mass spectrometry, we identified a protein, chitinase-like 3 (Chil3). Chil3 messenger RNA (mRNA) expression showed temporal changes, which included a peak increase at 2 days after injury. Next, we produced an anti-Chil3 antibody and confirmed its specificity with western blotting. Similar to the mRNA results, an analysis of temporal changes in Chil3 protein expression revealed a peak at 2 days after injury. We also investigated the time course of changes in Chil3 tissue localization using immunohistochemistry. Chil3 signals remained in the wounded area for up to 9 days. However, Chil3-positive cells were observed in the scab, the edge of the dermal layer, and neogenetic granulation tissue between 1 and 3 days. Thus, wound age can be histologically determined using the localization of Chil3 but not its general existence. Additionally, double-labeled fluorescent immunohistochemistry revealed that the Chil3-expressing cells were mainly neutrophils. These data show that Chil3 is expressed in neutrophils during the early stage of wound healing in mice; thus, Chil3 is a potential histological marker of 1-3-day-old wounds.

Immunohistochemical detection of CD14 and combined assessment with CD32B and CD68 for wound age estimation.

Estimation of wound age is a major topic of study for forensic pathologists, but few markers exist that can indicate a specific period 1-5 days postinfliction, and a method to estimate wound age with high accuracy has not yet been established. This study examined CD14 as such a marker in mouse skin wounds of different ages (0min and 1, 2, 3, 5, 7, and 9 days) and in human subjects (group 1, 0-1 day; group 2, 1-5 days; group 3, >7 days) using Western blot analysis and/or immunohistochemical staining. In addition, we evaluated a combination of immunohistochemical markers in human skin wounds using transmembrane proteins, CD14, CD32B, and CD68, expressed on inflammatory cells. The expression of CD14 was detected only during 1-5 days postinfliction and, thus, the evaluation of CD14-expressing cells could specify wound age during 1-5 days postinfliction in mouse skin wounds. The ratio of samples assessed to be CD14(+) was significantly high in human skin wounds in group 2. Combined assessment using the three markers increased the specificity of diagnosis and shortened the range of wound age, compared with the assessment using a single marker. Our results indicate that CD14 may be a useful marker of wound age, 1-5 days postinfliction, and that combined assessment with CD14, CD32B, and CD68 may be a good method for the accurate estimation of wound age.

Use of Aquamid as a filler for facial rejuvenation in orientals.

We used Aquamid as a filler for facial augmentation and rejuvenation in Orientals. This article introduces the injection techniques, effects, adequate dosage and complications of this filler, especially about rejuvenation of nasolabial fold and nasojugal groove. From December 2002 to June 2007, 5676 patients were treated in our clinic group. Complications were relatively minimal (0.082%) in comparison to other fillers and long-term effects were revealed. This is the first report concerning Aquamid use in facial rejuvenation of the Orientals.

The time-course analysis of gene expression during wound healing in mouse skin.

RNA analysis has been applied to forensic work to determine wound age. We investigated mRNA expression using quantitative RT-PCR of ten genes, including c-fos, fosB, mitogen-activated protein kinase phosphatase-1 (MKP-1), CD14, chemokine (C-C motif) ligand 9 (CCL9), placenta growth factor (PlGF), mast cell protease-5 (MCP-5), growth arrest specific 5 (Gas5), beta-2 microglobulin (B2M) and major urinary protein-1 (MUP-1), in terms of repair response in adult mice. The expression level of c-fos, fosB and MKP-1 transcripts increased drastically, peaked within 1h, and that of the CD14 and CCL9 transcripts peaked from 12 to 24h. An increase in PlGF and MCP-5 mRNA appeared on about day 5. Gas5, B2M and MUP-1 transcripts showed no significant change. Each gene had differentially expressional patterns with time-course. Our result implied that the observation of the 7 genes in wounded skin could serve to aid in the accurate diagnosis of wound age.

Bednar tumor: a report of two cases.

We have recently treated two Japanese with Bednar tumor, pigmented dermatofibrosarcoma protuberans. The tumors were multilobular with brown-pigmentation and an irregular surface. Firm nodules were growing into the deep subcutaneous tissue, but they were movable over the underlying tissue. Histopathological examination and immunohistochemical studies showed the typical features of Bednar tumor and positive staining for CD34, which is known to be expressed in dermatofibrosarcoma protuberans. Bednar tumor is thought to be a variant of dermatofibrosarcoma protuberans and to have intermediate malignancy.