凉血解毒汤对银屑病小鼠皮肤组织CCL20/CCR6表达的影响

目的:观察凉血解毒汤对银屑病小鼠皮肤组织中CC趋化因子配体20(CCL20)/CC趋化因子受体6(CCR6)表达的干预作用。方法:24只BALB/c雄性小鼠随机分为空白对照组、银屑病模型组、凉血解毒汤中药治疗组和趋化因子CCL20单克隆抗体(阳性药)治疗组。用咪喹莫特诱导小鼠银屑病模型,采用银屑病皮损面积和疾病严重程度(psoriasis area and severity index,PASI)评分标准观察银屑病样小鼠皮损的变化情况。光镜下观察皮损组织形态学变化,测量表皮层厚度。采用real-time PCR检小鼠皮肤组织样本中CCL20和CCR6表达的变化。结果:银屑病模型组小鼠皮肤出现大量鳞屑和红斑,表皮增厚;与银屑病模型组小鼠比较,凉血解毒汤组小鼠银屑病样皮损程度较轻,红斑、鳞屑以及表皮增厚程度轻于模型组,PASI分数降低,皮肤组织CCL20以及CCR6的表达明显低于模型组。趋化因子CCL20单克隆抗体治疗组小鼠银屑病皮损程度较轻,红斑、鳞屑以及表皮增厚程度轻于模型组,PASI分数降低,皮肤组织CCL20以及CCR6的表达明显低于模型组。结论:凉血解毒汤可能通过下调CCL20/CCR6的表达来减轻银屑病小鼠的皮肤病变。     

The Vitamin D Analogue Calcipotriol Reduces the Frequency of CD8+IL‐17+ T Cells in Psoriasis Lesions

The vitamin D analogue calcipotriol is an immunomodulatory drug widely used to treat psoriasis; however, how calcipotriol affects the immune cells in psoriasis lesions is not fully understood. The aim of this atudy was to investigate the effect of calcipotriol on the frequency of CD4⁺ and CD8⁺ T cells and innate lymphoid cells (ILC) and their production of IL‐17A, IFN‐γ and IL‐22 in psoriasis lesions in patients with chronic plaque psoriasis. Eighteen patients with psoriasis were included, and two similar psoriasis lesions were chosen for each patient. One lesion was treated with calcipotriol (50 μg/g) and the other with vehicle twice a day for 14 days. The clinical effect was measured by degree of erythema, scaling and induration in each lesion (SUM score). Skin biopsies were collected for histological and immunohistochemical analyses. Skin‐derived cells were isolated and analysed by flow cytometry. After 14 days of treatment with calcipotriol, a significant clinical and histological effect was seen; however, we found no differences in the frequency of CD4⁺ and CD8⁺ T cells or ILC between calcipotriol‐ and vehicle‐treated skin. The main finding was a significant decrease in CD8⁺IL‐17⁺ T cells in skin‐derived cells from calcipotriol‐treated skin, which was further supported by the absence of CD8⁺IL‐17⁺ T cells in immunohistochemical staining of calcipotriol‐treated skin. No changes in the frequency of IL‐22⁺ or IFN‐γ⁺ cells were observed. Our findings show that the vitamin D analogue calcipotriol reduces the frequency of CD8⁺IL‐17⁺ T cells in psoriasis lesions concomitant with clinical improvement.     

3D surface roughness measurement for scaliness scoring of psoriasis lesions

Psoriasis is an incurable skin disorder affecting 2–3% of the world population. The scaliness of psoriasis is a key assessment parameter of the Psoriasis Area and Severity Index (PASI). Dermatologists typically use visual and tactile senses in PASI scaliness assessment. However, the assessment can be subjective resulting in inter- and intra-rater variability in the scores. This paper proposes an assessment method that incorporates 3D surface roughness with standard clustering techniques to objectively determine the PASI scaliness score for psoriasis lesions. A surface roughness algorithm using structured light projection has been applied to 1999 3D psoriasis lesion surfaces. The algorithm has been validated with an accuracy of 94.12%. Clustering algorithms were used to classify the surface roughness measured using the proposed assessment method for PASI scaliness scoring. The reliability of the developed PASI scaliness algorithm was high with kappa coefficients>0.84 (almost perfect agreement).     

Experimental and numerical study of the colour appearance of tattoo models

The colour of tattooed skin has been predicted by a Monte Carlo method based on the optical coefficient spectra of the skin and tattoo dyes. Slices of pig skin, a tattoo phantom and skin phantoms with different thickness were prepared, and their reflectance and transmittance spectra were measured using an integrating sphere at wavelengths varying from 400 nm to 700 nm. The absorption and scattering coefficient spectra of skin phantoms, pig skins and the tattoo phantom were each calculated using the inverse Monte Carlo method. The skin phantoms and pig skins were overlaid on the tattoo phantom, and the reflectance spectra of the two-layered structures were measured. The reflectance spectra of the two-layered structures were calculated from the optical coefficient spectra using the Monte Carlo method. They agreed well with the measured spectra. The colour differences between the calculated and measured spectra were also evaluated by the L^*a^*b^* colour space distances and showed good agreement, with 3.49 for the skin phantoms and 8.27 for the pig skins.     

Increased Th17 cells are accompanied by FoxP3 Treg cell accumulation and correlated with psoriasis disease severity

Both T-helper 17 cells (Th17) and CD4⁺CD25⁺ regulatory T cells (Treg) play important roles in the pathogenesis of psoriasis. However, the relationship between Th17 and Treg cells and their dynamic variations in psoriasis remain unclear. In this study, we found that both Th17 and FoxP3⁺ Treg cells were increased in psoriasis patients both in the peripheral circulation and skin tissue lesions and were positively correlated with disease severity. The ratio of Th17 to Treg cells in skin tissue lesions was inversely correlated with PASI scores, while it was positively correlated with PASI scores in the circulation. IL-17 secretion by CD4⁺ T cells was not regulated by Treg cells, even though Treg cells exhibited significant inhibition on CD4⁺ T cells proliferation and IFN-γ production. These findings provide new information regarding the association between Th17 and Treg cells, which will further our understanding of the pathogenesis of psoriasis.     

Association of Disease Severity with IL‐1 levels in Methotrexate‐treated Psoriasis Patients

Interleukin‐1 plays a key role in inflammation and keratinocyte activation. It is an important mediator in the initiation and maintenance of psoriatic plaques and may represent an attractive therapeutic target. The aim of this study is to evaluate the effect of Methotrexate (MTX) on IL‐1 α and IL‐1 β levels in both plasma and skin biopsy of patients with psoriasis and to investigate their association with clinical disease activity. Forty‐five control subjects and 58 patients with psoriasis were recruited for this study. The patients were treated with 7.5 mg of MTX per week for 12 weeks. Folic acid was given at 5 mg once daily except on the day of MTX for 12 weeks. Blood samples and lesional skin biopsy were taken. Disease severity was assessed by Psoriasis Area Severity Index (PASI) score. IL‐1 levels in plasma and skin biopsy were analysed using ELISA. PASI score declined significantly (P < 0.001) from day 0 to 12 weeks of MTX treatment. IL‐1 α level in plasma and skin biopsy was reduced at day 0 sample and elevated significantly (P < 0.001) after MTX treatment. IL‐1β level in plasma and skin biopsy was higher at day 0 sample and reduced significantly (P < 0.001) after MTX treatment. IL‐1α levels and PASI score showed inverse correlation score before and after treatment with MTX. Whereas IL‐1β levels showed positive correlation before and after treatment with MTX. Decreasing IL‐1β levels by MTXs in psoriasis may block the Th17 differentiation. This shows the therapeutic effect of MTX in controlling the immunopathogenesis of psoriasis.     

Effects of Expression of Transcriptional Factor AP-1 <Emphasis Type="Italic">FOSL1</Emphasis> Gene on Psoriatic Process

We performed quantitative analysis of FOSL1 gene expression in lesional psoriatic skin. The expression of this gene in lesional psoriatic skin was significantly increased compared to that in unaffected areas. Enhanced FOSL1 expression significantly correlated with high psoriasis area and severity index (PASI). High level of FOSL1 gene expression was proposed to be a marker of pathological process activity in psoriasis.     

Nuclear Factor Kappa B and Cyclo-Oxygenase-2: Two Concordant Players in Psoriasis Pathogenesis

Nuclear factor kappa B (NFκB) is a key regulatory element in a variety of immune and inflammatory pathways, cellular proliferation, differentiation and apoptosis. Cyclo-oxygenase 2 (COX2) is one of the downstream targets of NFκB. The current work aimed to explore the possible role of NFκB and COX2 in psoriasis pathogenesis through their immunohistochemical (IHC) expression in skin biopsies of this disease and correlating this expression with clinico-pathological parameters of studied cases. 103 subjects were studied; including 58 cases with psoriasis vulgaris (lesional and perilesional skin) and 45 normal, age- and gender-matched subjects, as a control group. NFκB and COX2 expressions were evaluated using standard IHC techniques. NFκB and COX2 were upregulated in psoriasis lesional skin compared to perilesional (p?p?p?=?0.02 for both), severe degree of perivascular inflammatory infiltrate (p?=?0.03 and 0.002, respectively) and thin suprapapillary epidermis (p?=?0.003 and 0.006, respectively). Significant positive correlation was noted between NFκB and COX2 H scores in epidermis (r?=?0.41, p?=?0.02) and dermis (r?=?0.6, p?=?0.04) of lesional skin. Significant positive correlation between NFκB H score and PASI score (r?=?0.38, p?=?0.04) and between COX2 H score and PASI score (r?=?0.52, p?相似文献   

Evidence for a role of autoinflammation in early-phase psoriasis

Psoriasis is a common, inflammatory immune-mediated skin disease mainly presenting with plaques whose pathogenesis is based on the central role of the interleukin (IL)-23/IL-17 axis. However, the mechanisms acting in papular lesions of early-phase psoriasis are not fully understood. The aim of this study was to assess the involvement of autoinflammation, a state of sterile inflammation mainly driven by IL-1 over-production that has been recently hypothesized to act in the early phase of disease. Lesional skin of 10 patients with recent onset, untreated psoriasis has been investigated for expression of IL-1β, IL-17, IL-23 and other cytokines involved in the disease in comparison with normal skin of 10 healthy controls using a protein array method. Immunohistochemical phenotyping of inflammatory infiltrate and co-localization experiments with immunofluorescence confocal microscopy were conducted. IL-1β was significantly more expressed in psoriasis than in normal skin (P < 0·0001). The chemokine IL-8 was also over-expressed in psoriasis (P = 0·03) while IL-12, IL-17, IL-23, tumour necrosis factor-α and interferon-γ were only slightly more expressed in psoriasis than in normal skin, without reaching statistical significance. The inflammatory infiltrate consisted mainly of neutrophils with a relevant number of macrophages and dendritic cells and only scattered, predominantly T helper 1 lymphocytes. IL-1β co-localized mainly with CD66b, a neutrophil marker, suggesting that neutrophils were the major source of this cytokine. IL-1β over-expression in combination with low expression of cytokines that are predominant in late-phase plaque psoriasis may support the role of autoinflammation in early-phase disease, possibly paving the way to randomized trials with IL-1 antagonists.     

Lesion area assessment in psoriasis patients

Psoriasis is an autoimmune skin disease, afflicting skin with red plaques that are usually accompanied by silvery-white scales. Various medical treatments are used, with different impacts on the patients, but there is no definite cure for the disease. The PASI standard is employed to measure the performance of the treatments. It includes four parameters, namely area, erythema, scaliness and skin thickness. The PASI parameters are usually measured manually by physicians through subjective clinical observations which are imprecise, time consuming and in some cases lead to diverse results. This paper presents a computer-based automatic method to measure the area parameter in the PASI standard. In the proposed method, the YCbCr colour space is used to differentiate the plaques from the skin by applying an optimal threshold method. Performance evaluation results indicate that the proposed method is able to determine lesion areas with accuracy higher than 96% for 18 out of 20 cases and higher than 92% for another case. As well as high accuracy the proposed method has another advantage over previous methods: it can automatically detect plaques with silvery-white scales, plaques on hairy skins and tiny plaques, as well as simple (scale-less) plaques.     

Lesion area assessment in psoriasis patients

Psoriasis is an autoimmune skin disease, afflicting skin with red plaques that are usually accompanied by silvery-white scales. Various medical treatments are used, with different impacts on the patients, but there is no definite cure for the disease. The PASI standard is employed to measure the performance of the treatments. It includes four parameters, namely area, erythema, scaliness and skin thickness. The PASI parameters are usually measured manually by physicians through subjective clinical observations which are imprecise, time consuming and in some cases lead to diverse results. This paper presents a computer-based automatic method to measure the area parameter in the PASI standard. In the proposed method, the YCbCr colour space is used to differentiate the plaques from the skin by applying an optimal threshold method. Performance evaluation results indicate that the proposed method is able to determine lesion areas with accuracy higher than 96% for 18 out of 20 cases and higher than 92% for another case. As well as high accuracy the proposed method has another advantage over previous methods: it can automatically detect plaques with silvery-white scales, plaques on hairy skins and tiny plaques, as well as simple (scale-less) plaques.     

Pathogenic function of IL-1 beta in psoriasiform skin lesions of flaky skin (fsn/fsn) mice

IL-1 acts on many cells as an inflammatory mediator. Its two forms, IL-1α and IL-1β, are regulated differentially within hyperproliferative inflammatory skin conditions, such as psoriasis. While IL-1α is down-regulated within psoriatic lesions, the levels of IL-1β are increased. However, some investigators have described an inactive form of IL-1β in psoriasis, while others have detected increased IL-1β activity within these lesions. Thus, its in vivo role remains unclear. We have assessed expression and function of IL-1β within psoriasiform skin lesions of the spontaneous mouse mutation flaky skin (fsn/fsn). It was found that IL-1β was increased by 357% within psoriasiform lesions of fsn/fsn mice compared with their wild-type or heterozygous (+/?) littermates (P < 0·00001). When the IL-1β function was inhibited by i.p. injection with a neutralizing MoAb, no effects were seen in +/? mice. In contrast, psoriasiform features in fsn/fsn mice were alleviated dramatically, as demonstrated by a 40% decrease of the epidermal thickness and a diminished number of intra-epidermal microabscesses. In addition, infiltrating epidermal CD4⁺ and CD8⁺ T cells were decreased by 68% and 81%, respectively (P < 0·05), and epidermal Langerhans cells also were reduced by 36% (P < 0·005). In contrast, mast cells were not affected, suggesting differential responses of various cutaneous cell types. Our results demonstrate an important in vivo role of IL-1β for the generation of hyperproliferative inflammatory skin lesions in the fsn/fsn model.     

Plexin-B2 in psoriasis; a clinical and immunohistochemical study

ABSTRACT

Psoriasis is an inflammatory, immune-mediated disease. Plexins are transmembrane proteins that are involved in immune system regulation and inflammation. This work aimed to investigate the immunohistochemical expression of Plexin-B2 in plaque psoriasis in both lesional and perilesional skin. This case‐control study included 30 patients with psoriasis vulgaris in comparison with 20 age- and sex‐matched apparently healthy persons. We used the Psoriasis Area and Severity Index (PASI) score to evaluate psoriasis severity. Biopsies from 30 lesional, 30 perilesional, and 20 control-skin patients were subjected to histopathological and immunohistochemical evaluations of Plexin-B2. There was significant stepwise overexpression of Plexin-B2 in proliferating keratinocytes from controls (66 ± 31.02) to perilesional (116 ± 41.95) and lesional (159.7 ± 63.05) skin (P < .001). Also, Plexin-B2 showed significant overexpression in dermal inflammatory cells of lesional psoriatic skin (153.67 ± 72.71) when compared to controls skin (25.71 ± 11.34) (P < .001). There was a significant positive correlation between Plexin-B2 expression and psoriasis severity (r = 0.557; P < .001). Plexin-B2 could promote skin inflammation, as well as keratinocyte proliferation in psoriasis vulgaris; therefore, it may be used as a targeted therapy for psoriasis treatment.     

Impact of clinical variables on Borrelia burgdorferi-specific antibody seropositivity in acute-phase sera from patients in North America with culture-confirmed early Lyme disease

Erythema migrans, the most common manifestation of Lyme disease, has been associated with highly variable rates of seropositivity for antibodies to Borrelia burgdorferi. Differences in the sensitivities of serologic assays for the detection of these antibodies, however, may not be the only or even the primary explanation for this observation. We investigated the impacts of four clinical variables on seropositivity—the duration of erythema migrans, the presence of single versus multiple skin lesions, and the gender and age of the patient. In this analysis, three different serologic tests were performed on acute-phase sera from 175 untreated patients with culture-confirmed erythema migrans: the C6 single-peptide enzyme-linked immunosorbent assay (ELISA), a commercially available ELISA in which a whole-cell sonicate of B. burgdorferi was the antigen, and a two-tier procedure. Irrespective of the serologic test performed, the results showed that seropositivity rates increased with the duration of the erythema migrans for patients with single lesions (P < 0.001) but not for those with multiple skin lesions. The variability in seropositivity rates was greatest for the two-tier testing strategy, with a >6-fold-higher rate of seropositivity among patients with a single lesion of 22- to 30-day duration than among those whose skin lesion was of 1- to 7-day duration (85.7 versus 14.1%; P < 0.001). Rates of seropositivity by each of the testing methods were also significantly higher for patients with multiple skin lesions than for those with single lesions (P < 0.001). In contrast, seropositivity rates were not affected by either the gender or the age of the patient. Thus, in patients with erythema migrans, certain clinical variables such as the duration and number of skin lesions had a profound impact on seropositivity rates, irrespective of the serologic assay performed.     

Clinical,histologic, and electron microscopy study of skin exposed to low‐frequency ultrasound

The use of low‐frequency ultrasound has been proposed to enhance the transdermal transport of various drugs, a technique referred to as sonophoresis. The aim of the present study was to determine the safety of low‐frequency sonophoresis on human and rat skin by evaluating their structural modifications after ultrasound exposure. Human skin samples and hairless rats were exposed to 20 kHz ultrasound in vitro and in vivo, respectively. Ultrasound was used with average intensities ranging from 0.25 to 7 W/cm² in pulsed or continuous mode. Hairless rats were also exposed to a heat source mimicking the temperature versus time profile during sonication. Skin samples were observed under optical and electron microscopy to detect any structural changes. Human skin samples exposed to intensities lower than 2.5 W/cm² showed no modification. For hairless rats, slight and transient erythema was observed after 2.5 W/cm² exposure, whereas deep lesions (dermal and muscle necrosis) were observed 24 hr later. These lesions were also observed when a plastic film was placed between the coupling medium and the animals' skin during sonication. In contrast, no histologic lesion could be seen when a heat source was applied to animal skin. Low‐frequency ultrasound induces delayed and deep lesions in hairless rat skin at 2.5 W/cm² which are not only attributable to the increase in temperature at the skin surface during ultrasound exposure. By using the same ultrasound conditions, human skin seems to be less sensitive in vitro. Anat Rec 264:114–119, 2001. © 2001 Wiley‐Liss, Inc.     

Evaluation of Aquaporin-3 Role in Nonmelanoma Skin Cancer: An Immunohistochemical Study

Aquaporin-3 (AQP3), is an aquaglyceroporin, that plays a role in cell proliferation, tumorigenesis, and cell migration. This study aimed at evaluating the possible role of AQP3 in nonmelanoma skin cancer (NMSC) pathogenesis through its immunohistochemical expression in skin biopsies of these diseases. One-hundred and thirty cutaneous specimens were studied. These included 60 cases of NMSC and 40 normal skin and 30 psoriasis samples, from age- and gender-matched subjects, as a control group. AQP3 was expressed in 66.7% of basal cell carcinoma (BCC) cases and in 93.3% of squamous cell carcinoma (SCC) cases. Higher AQP3 expression (p?=?.01), expression percentage (p?=?.01), and H score (p?=?.04) were significantly associated with SCC compared to BCC. Normal skin and psoriasis showed significantly higher AQP3 expression (p?=?.001, p?<?.001, respectively), expression percentage (p?<?.001 for both), and H score values (p?<?.001, p?=?.001, respectively) compared to NMSC. Higher H score values in BCC were significantly associated with female gender (p?=?.02) and with nodular lesions (p?>?.001). Higher H score values in SCC were significantly associated with grade III tumors (p?=?.04) and AQP3 percentage of expression was significantly correlated with grade III tumors (r?=?.48, p?=?.009). In conclusion, AQP3 may play a role in NMSC pathogenesis. This probably occurs through aquaporin-mediated glycerol transport and ATP generation. Its downregulation, observed in the current work, is mostly a result of excessive proliferation. Further studies are needed to investigate the therapeutic effect of its inhibition in NMSC treatment.     

Performance of a dermoscopy-based computer vision system for the diagnosis of pigmented skin lesions compared with visual evaluation by experienced dermatologists

BackgroundIt is often difficult to differentiate early melanomas from benign melanocytic nevi even by expert dermatologists, and the task is even more challenging for primary care physicians untrained in dermatology and dermoscopy. A computer system can provide an objective and quantitative evaluation of skin lesions, reducing subjectivity in the diagnosis.ObjectiveOur objective is to make a low-cost computer aided diagnostic tool applicable in primary care based on a consumer grade camera with attached dermatoscope, and compare its performance to that of experienced dermatologists.Methods and materialsWe propose several new image-derived features computed from automatically segmented dermoscopic pictures. These are related to the asymmetry, color, border, geometry, and texture of skin lesions. The diagnostic accuracy of the system is compared with that of three dermatologists.ResultsWith a data set of 206 skin lesions, 169 benign and 37 melanomas, the classifier was able to provide competitive sensitivity (86%) and specificity (52%) scores compared with the sensitivity (85%) and specificity (48%) of the most accurate dermatologist using only dermoscopic images.ConclusionWe show that simple statistical classifiers can be trained to provide a recommendation on whether a pigmented skin lesion requires biopsy to exclude skin cancer with a performance that is comparable to and exceeds that of experienced dermatologists.     

Psoriasis in humans is associated with down-regulation of galectins in dendritic cells

We have investigated the expression and role of galectin‐1 and other galectins in psoriasis and in the Th1/Th17 effector and dendritic cell responses associated with this chronic inflammatory skin condition. To determine differences between psoriasis patients and healthy donors, expression of galectins was analysed by RT‐PCR in skin samples and on epidermal and peripheral blood dendritic cells by immunofluorescence and flow cytometry. In the skin of healthy donors, galectin‐1, ‐3 and ‐9 were expressed in a high proportion of Langerhans cells. Also, galectins were differentially expressed in peripheral blood dendritic cell subsets; galectin‐1 and galectin‐9 were highly expressed in peripheral myeloid dendritic cells compared with plasmacytoid dendritic cells. We found that non‐lesional as well as lesional skin samples from psoriasis patients had low levels of galectin‐1 at the mRNA and protein levels, in parallel with low levels of IL‐10 mRNA compared with skin from healthy patients. However, only lesional skin samples expressed high levels of Th1/Th17 cytokines. The analysis of galectin‐1 expression showed that this protein was down‐regulated in Langerhans cells and dermal dendritic cells as well as in peripheral blood CD11c⁺ DCs from psoriasis patients. Expression of galectin‐1 correlated with IL‐17 and IL‐10 expression and with the psoriasis area and index activity. Addition of galectin‐1 to co‐cultures of human monocyte‐derived dendritic cells with autologous T lymphocytes from psoriasis patients attenuated the Th1 response. Conversely, blockade of galectin binding increased IFNγ production and inhibited IL‐10 secretion in co‐cultures of monocyte‐derived dendritic cells with CD4⁺ T cells. Our results suggest a model in which galectin‐1 down‐regulation contributes to the exacerbation of the Th1/Th17 effector response in psoriasis patients. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Optical devices used for image analysis of pigmented skin lesions: a proposal for quality assurance protocol using tissue-like phantoms

Different technological tools have been developed to aid in the diagnosis of pigmented skin lesions, including cameras working with conventional RGB colour systems, epiluminescence microscopy and spectrophotometric methods using visible and near infrared wavelengths. All the different procedures should provide in an objective and reproducible fashion quantitative measurements of the colour and shape features of a given skin mole. At present, many devices have been introduced in experimental stages for clinical diagnosis, mainly used to provide to the clinicians an objective, computer-assisted second opinion. As for any diagnostic instruments, optical devices should also be subjected to a dedicated quality assurance protocol in order to evaluate the response repeatability of each device (intra-instrument agreement) and to check the accordance among the responses of different devices (inter-instrument agreement). The aim of this study was to design a quality assurance protocol for optical devices dedicated to image analysis of pigmented skin lesions and, in case, to detect cutaneous melanoma by using suitable tissue-like phantoms as standard references that enable testing of both hardware and software components. As an example, we report the results of intra-instrument and inter-instrument agreement when the protocol was applied on a series of 30 SpectroShade instruments, a novel optical device based on multi-spectral image analysis of colour and shape features of pigmented skin lesion.