| Objective:HE staining and immunohistochemical examination were made for pathological specimens of nevus flammeus in different color types,in order to explore the morphological differences of Port Wine-stains (PWS) in different clinical manifestations, and make better choice for laser parameters in clinical treatment and improve the therapeutic effect. The tissue pathology of different types were observed through optical microscope and the pathological images were analysised and measure by computer, including mean vessels diameter, number density, mean vessels depth and the maximum depth of the blood vessels.Method:Patients with PWS were choosed during march 2008 to march 2009, all patients were divided into four types by the color of lesion area according to combing visual observation of two or more physician with spectrophotometric color measurement form CM-2600d. These were pink type (typeⅠ), red type (typeⅡ), purple type (typeⅢ) and purple with hypertrophic nodularity type(typeⅣ).10 cases were selected randomly in each type which were not accepted any treatment before and all lesions were in the face or neck. In addition,10 cases of healthy skin tissues in the face or neck were selected randomly from the 40 patients. After local anesthesia by 2% lidocaine, the focus and the corresponding normal tissues were collected by scalpel. After regular fixation and embedding, they were sliced up, and the prepare sections were stained with HE and marked the vessels with geneⅧfor immunohistochemistry.The slices stained with HE were put under light microscopes to observe the pathology of the tissues, especially the distribution of the malformed blood vessels. The slices stained with immunohistochemistry underwent a computed image assessment, and measured the following parameters:the mean vessels diameter, the mean vessels depth, the maximum depth, and figure out the number density based on principles of stereology. Statistic processing "analyzed by the SPSS16.0, the experimental data were figured as (x±s).The margin of P<0.05 had statistic significances after the ANOVA, exact probability test and rank sum test of the materials.Results:Histological examination:the cuticle was normal with the stain of HE, and the capillary vascular displayed dilatation with groups in the superficial dermis. With darkening of the color of the pathological changes, the number of the vessels increasing and the diameter enlarging, part of the dilatation capillary vascular extended to the deep dermis and tissues under the skin. The number of capillary vascular full of red cells were increasing. The endothelial cells matured but were not hypertrophic. Immunohistochemical examination:the brown capillary vascular in the superficial and middle dermis were abundant after the blood vessels marked with geneⅦwere stained positively. Most of them were dilated vessels. Computed image assessment:the mean vessels diameter (μm) of pink type, red type, purple type and purple with hypertrophic nodularity type were 55.39±2.74,94.55±2.13,133.85±11.51,171.07±13.75;The mean vessels depths (mm) were 0.40±0.10,0.43±0.21,0.52±0.08,0.62±0.12; The maximum depth (mm) were 0.51±0.07,0.61±0.16,0.72±0.11,0.87±0.18; The number density (/mm2) were 15.87±0.07,19.14±1.09,22.39±3.10,25.33±2.84;Capillary diameter of the normal control group (μm) was 14.23±0.85, the maximum depth (mm) was 0.51±0.17, the mean depth (mm) was 0.40±0.14, the number density (/mm2) was 6.70±0.34. Among these, mean vessels diameter and the number density had statistic differences between groups with different clinical symptoms. While the maximum depth between the pink type with the normal group, the mean vessels depths between the normal group and the pink type, the normal group and red type, the pink type and the red type had no statistic differences. And other clinical groups were all had significantly different.Conclusion:The patients'age increasing with the deepening of colour in focals area, meanwhile, the mean vessels diameter, the number density, the mean vessels depth and the maximum depth in focal ares also increasing gradually. The vascular distribution in focal areas of the pink type and the red type are relatively shallow, mainly presented for the expansion of blood vessels and increasement of the number;The distribution of dilated vessels in purple type and purple with hypertrophic nodularity type were deeper than the other types. The number of the vessels in typeⅢand typeⅣis increasing.With the color deepening of lesions, it can choose different wavelengths of laser, combined with increasing the pulse width and energy density properly.
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