Submicron resolution techniques: Multiphoton microscopy in skin disease.

Non-invasive optical examination plays a crucial role in various aspects of dermatology, such as diagnosis, management and research. Multiphoton microscopy uses a unique submicron technology to stimulate autofluorescence (AF), allowing for the observation of cellular structure, assessment of redox status and quantification of collagen fibres. This advanced imaging technique offers dermatologists novel insights into the skin's structure, positioning it as a promising 'stethoscope' for future development in the field. This review provides an overview of multiphoton microscopy's principles, technology and application in studying normal skin, tumour and inflammatory diseases, as well as collagen-related and pigmentary diseases.

Superficial basal cell carcinoma mimicking Bowen's disease: Diagnosis with noninvasive imaging.

BACKGROUND: Superficial basal cell carcinoma (SBCC) is the rare subtype of basal cell carcinoma (BCC). BCC occurs in exposed areas such as the head and face, SCBB prone to form in trunk. Due to the manifestation of erythema and desquamation, it is prone to misdiagnosed as Bowen's disease in clinica. MATERIALS AND METHODS: A 68-year-old female presented with coin-sized erythema located on the lower abdomen for 5 years. Histopathological examination was performed, and results informed the diagnosis of SBCC. Lesions were detected by dermoscopy, reflectance confocal microscopy (RCM) and multiphoton microscopy (MPM). RESULTS: Dermoscopy revealed yellow-red background with more dendritic and linear proliferating vessels and more blue-gray nonaggregated dots structures. RCM displayed streaming of stratum spinosum, tortuous dilated vessels, highlighted inflammatory cells, and medium refraction round and oval tumor cell masses. MPM showed epidermal cells in polar arrangement, increased cell spacing, disorganized stratum granulosum and elastic fibers are gathered in clusters. CONCLUSION: We described a case of SBCC detected by dermoscopy, RCM and MPM. Noninvasive imaging features may provide a potentially tools in recognition and differentiation of SBCC.

In vivo detection of healthy skin: Comparison of multiphoton microscopy and reflectance confocal microscopy.

BACKGROUND: Noninvasive skin examination evolved rapidly in recent years, with multiphoton microscopy (MPM) and reflectance confocal microscopy (RCM) being used to image in-vivo skin at high resolution. The aim of this study is to compare the imaging clarity between the two techniques and measure the thickness of the epidermis in different body sites. We also measured the degree of skin aging with noninvasive tools. METHODS: Fifty-six volunteers were evaluated and measured at three different body sites, including the cheek, volar forearm, and back. We used RCM and MPM to evaluate the clarity of each skin layer, including stratum corneum, stratum granulosum, stratum spinosum, dermo-epidermal junction, and dermis. We measured epidermal thickness (ET) at the three body sites in individuals of different ages and genders. We assessed skin aging by the second harmonic to autofluorescence aging index of dermis (SAAID), and multiple linear regression was used to analyze the factors affecting SAAID. RESULTS: MPM had advantages in observation of stratum granulosum, collagen fiber, and elastic fiber (p < 0.001), but RCM provided better observation in dermo-epidermal junction layer (p < 0.001). The epidermis was thicker in the cheek area than the volar forearm and back in both RCM and MPM detection, and the average ET measured by MPM was lower than RCM. ET varied among the three body sites with significant differences (p < 0.05). ET was significantly lower at almost all sites in individuals above 40y (p < 0.05). SAAID decreased with age, and more rapidly in women. Cheeks have lower SAAID scores than other body sites. CONCLUSION: MPM and RCM provide noninvasive methods for imaging skin and each method has its own advantages. Epidermal thickness and SAAID correlated with age, gender, and different body sites. MPM could also assess the degree of skin aging, which could guide the clinical treatment of patients with diffferent ages and genders in the above body sites.

Ceramide synthase 6 antisense RNA 1 contributes to the progression of breast cancer by sponging miR-16-5p to upregulate ubiquitin-conjugating enzyme E2C.

Breast cancer (BC) is the most dangerous female mortality all over the world, described by unavoidable spread and metastaticity of BC cells. Increasing evidences verified that lncRNA play a major role in the tumorgenesis and development of BC cell. The purpose of this study is to investigate the roles of lncRNA ceramide synthase 6 antisense RNA 1 (CERS6-AS1) and ubiquitin-conjugating enzyme E2C (UBE2C) in BC and explore the regulatory association among miR-16-5p, CERS6-AS1, and UBE2C in BC. The CERS6-AS1 and UBE2C expression levels were determined by real time quantitative PCR in cell lines and tissues of BC. The function of CERS6-AS1 and UBE2C in the apoptosis, proliferation, and migration was confirmed by cell counting kit-8, Transwell, and flowcytometry tests. We performed tumor xenograft assay to validate the roles of CERS6-AS1 in vivo. The expression of UBE2C proteins was evaluated by Western Blot analysis. Moreover, the relationship among UBE2C, CERS6-AS1, and miR-16-5p was verified by luciferase report assay. It was found that CERS6-AS1 and UBE2C were meaningfully upregulated in BC, and knockdown of both CERS6-AS1 and UBE2C inhibited the BC cell proliferation and migration, whereas induced apoptosis. Mechanistically, CERS6-AS1 could facilitate BC progression by sponging miR-16-5p for upregulation of the UBE2C expression. The CERS6-AS1/miR-16-5p/UBE2C axis might be a prospective therapeutic target in the BC treatment by sponging miR-16-5p to upregulate UBE2C, which might contribute to the development of BC.

Clinical and Histopathological Characteristics of Tumors Arising in Seborrheic Keratosis: A Study of 1365 Cases.

OBJECTIVE: The aim of this study was to describe the clinical and histopathological characteristics of seborrheic keratosis (SK) with secondary tumor and identify associated factors. METHODS: This study was a monocentric retrospective case-control study describing the factors associated with the development of tumors in SK. A total of 1365 patients with SK were admitted to our department between 2018 and 2021. All of the histopathological specimens and medical records of these patients were reviewed. We compared the data of 36 patients with tumors arising in SK with those of the remaining 1329 patients with only SK, which were designated the control group. RESULTS: The incidence of tumors arising in SK that were biopsied was 2.6%. The tumor types included squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and keratoacanthoma (KA). Compared with other patients with SK, elderly patients with immunosuppression, patients presenting surrounding erythema or ulceration, and those with SK in exposed areas had an increased possibility of developing a secondary tumor. Tumors arising within SK have the histopathological features of ulceration, cytologic atypia, mitosis, "malignant horn", trabecular cell cords, solar elastosis and severe inflammatory infiltration in the dermis. CONCLUSION: SK can show malignant progression in a small fraction of SK cases that are biopsied. This study is the first report on the clinical and histopathological characteristics of tumors arising in SK and identifies relevant factors in an Asian population.

Expression of Heat Shock Protein 105 in Cutaneous Squamous Cell Carcinoma: Correlation with Clinicopathological Characteristics.

BACKGROUND: Heat shock proteins (HSPs), a group of heat stress proteins, are characterized by highly conserved properties. Malignant transformation is a cellular stress, and the expression of HSPs may be affected during this process. Heat shock protein 105 (HSP105) is a protective protein that has long been observed in many cancer types, but little attention has been given to cutaneous squamous cell carcinoma (CSCC). As such, the objectives of this study were to observe the expression of HSP105 on CSCC and evaluate its correlation with clinicopathological characteristics. METHODS: This retrospective study enrolled 60 patients with CSCC. The patients' clinical data, including sex, age, tumor location, tumor type, and degree of pathological differentiation, were collected. The expression of HSP105 was measured by Western blot and immunohistochemical staining. RESULTS: HSP105 expression was decreased in CSCC (HSCORE=0.65 (0.30, 1.98)) compared with normal skin (HSCORE=2.20 (1.50, 2.80)) (P<0.001). These results were consistent with the Western blot analysis. HSP105 immunostaining of Bowen disease (HSCORE=1.28 (1.08, 2.40)) revealed higher expression than in verrucous carcinoma (HSCORE=0.30 (0.23, 0.85)), keratoacanthoma (HSCORE=0.53 (0.29, 0.93)) and acantholytic squamous cell carcinoma (HSCORE=0.53 (0.41, 0.68) (P<0.01)). Poorly differentiated CSCC showed significantly higher expression of HSP105. CONCLUSION: Our study reveals for the first time that the expression of HSP105 is decreased in CSCC. We suggest that the molecular mechanisms underlying the differential expression of HSP deserve a more rigorous future study, the results of which might explain its role in carcinogenesis and its potential as a target for selective tumor therapy.

HSP105 expression in cutaneous malignant melanoma: Correlation with clinicopathological characteristics.

BACKGROUND: Heat shock proteins can protect against stress-associated cellular challenges, but they can also protect some tumors from human immune system monitoring. Heat shock protein 105 (HSP105/110) is a high molecular weight protein whose expression has been reported in many cancers, but few studies on its role in cutaneous malignant melanoma have been published. In this study, we analyzed the relationship between HSP105 expression and the clinicopathological characteristics of CMM. METHODS: This retrospective study included 91 patients with CMM. The clinicopathological characteristics of CMM patients, including age, lesion duration, location, pathological classification, Clark's level, Breslow thickness, metastasis and recurrence, were collected. Immunohistochemical staining and Western blot analysis for HSP105 were performed. Pigmented nevi (n = 20) served as a control. The staining intensity and percentage of stained cells were expressed as a histochemical score (HSCORE). RESULTS: HSP105 was overexpressed in melanoma compared with nevi. Differences in the HSCORE between nevi (HSCORE = 1.05(0.15,1.50)) and CMM (HSCORE = 2.68(1.80,3.60)) were remarkable (P<0.001). Exposed site lesions, recurrent and metastatic lesions, nodular melanoma and lentigo maligna melanoma were closely associated with higher HSP105 expression (P = 0.011, P = 0.001 and P = 0.001, respectively). Moreover, no significant difference was observed in Clark's level, Breslow thickness, or lesion duration (P>0.05). CONCLUSION: HSP105 is overexpressed in CMM. Higher HSP105 expression in lesions is associated with different clinicopathological variables. HSP105 may be a potential target for the diagnosis, treatment and prognostic prediction of CMM.

Bullous Scabies: Clinical, Dermoscopic, and Pathologic Characteristics of Ten Patients.

Bullous scabies (BS) is a rare atypical clinical variant of scabies and is easily confused with bullous disorders. The diagnosis of BS is always a challenge, and physicians often misdiagnose BS patients. Patients with BS admitted from 2012 to 2020 were enrolled in this study. The clinical, dermoscopic, and pathological characteristics of the patients were analyzed retrospectively. Ten patients with BS were enrolled in this study. Seven of the 10 patients were male. The bullae were most commonly found on the thighs and arms (80% of patients). Only 30% of patients (3/10) tested positive for mites and/or eggs by the initial skin scraping, but 100% (5/5) of the patients who received dermoscopy tested positive. Among these 10 patients, only five received a skin biopsy. Subepidermal (4/5) and intraepidermal (1/5) bullae with eosinophil and neutrophil infiltration were observed in five patients. Direct immunofluorescence (DIF) indicated linear deposition of IgG in the basement membrane zone in three patients. Physicians should consider the possibility of BS in patients with blisters, pruritus, and poor response to corticosteroids. Dermoscopy should be prioritized for the differential diagnosis of BS to exclude other bullous disorders. Finally, a biopsy should be performed on each patient with bullae.

The HSP90 Inhibitor, 17-AAG, Influences the Activation and Proliferation of T Lymphocytes via AKT/GSK3ß Signaling in MRL/lpr Mice.

OBJECTIVE: To explore the molecular mechanism of 17-AAG in the treatment of systemic lupus erythematosus (SLE), and the effects of the heat shock protein 90 (HSP90) inhibitor 17-AAG on the activation and proliferation of lymphocytes and the AKT/GSK3ß signaling pathway in MRL/lpr mice were detected. METHODS: MRL/lpr mice were randomly divided into the control group and the experimental group. The experimental group was injected intraperitoneally with 17-AAG, and T lymphocytes were separated by magnetic beads. Lymphocyte proliferation was detected by MTT and flow cytometry (FCM), and the expression of the HSP90 protein and PI3K/AKT signaling pathway-related proteins was detected by Western blotting. Renal histopathology and immune complex deposition were also observed in both groups. RESULTS: Immune complex deposition and inflammation decreased in kidneys from MRL/lpr mice in the experimental group. HSP90 protein expression, T lymphocyte proliferation and phosphorylated AKT and GSK3ß levels also decreased in the experimental group. CONCLUSION: 17-AAG can inhibit the activation and proliferation of T lymphocytes and downregulate the AKT/GSK3ß signaling pathway, which may be relevant for the treatment of SLE.

Flavonoids with iNOS inhibitory activity from Pogonatherum crinitum.

Pogonatherum crinitum has long been used as a folk remedy for the treatment of many inflammatory diseases in Taiwan, and till now there is still no report concerning its active principles as well as their pharmacological studies. That prompted us to investigate the bioactive constituents of Pogonatherum crinitum. Two novel chemical entities, luteolin 6-C-beta-boivinopyranoside (1) and 6-trans-(2''-O-alpha-rhamnopyranosyl)ethenyl-5,7,3',4'-tetrahydroxyflavone (2), along with luteolin (3), kaempferol (4), luteolin 6-C-beta-fucopyranoside (5), kaempferol 3-O-alpha-L-rhamnopyranoside (6), luteolin 6-C-beta-glucopyranoside (7), rutin (8) and kaempferol 3-O-rutinoside (9) were isolated from this plant, and identified by spectroscopic analysis. The effect of these compounds on the inhibition of NO production in LPS-activated macrophages was further evaluated. All these compounds inhibited NO production in activated RAW 264.7 cells to various degrees without affecting the cellular viability. Among the compounds examined, both compounds 1 and 2 suppressed LPS-induced NO production, with E(max) values of 99.51+/-0.23% and 92.41+/-3.22%, respectively. The most potent compounds, 3 and 4, inhibited NO production with IC(50) values of 10.41+/-0.02 microM and 10.61+/-0.44 microM, respectively. These effects were attributed to suppression of mRNA expression of inducible NO synthase (iNOS). Our results clearly demonstrated that these naturally occurring iNOS inhibitors may be beneficial to the treatment of inflammatory diseases associated with overproduction of NO, which provides an explanation, at least a part, for the anti-inflammatory property of Pogonatherum crinitum.

[Characteristics and influencing factors of air pollution in and out of the highway toll gates].

During June and July 2003, CO, NO2, THC and PM10 were sampled at the four highway toll gates in Chongqing. Air temperature, air pressure, wind velocity and traffic flow were also monitored simultaneously. The relation between air pollution parameters and influencing factors was analyzed by applying the methods of bivariate correlation and partial correlation. As shown in the monitoring result, the outdoor average concentrations of CO and PM10 exceed indoor ones, but NO2 and THC are reverse. The average concentrations of CO and NO2 at the toll gates don't exceed the indoor and outdoor air quality standards except for the toll gate in Chongqing and Chayuan. One-hour average concentrations of outdoor and indoor THC are 7.728 mg/m3 and 7.216 mg/m3 respectively, and exceed ten times of the indoor air quality standard. One-hour average concentrations of indoor and outdoor PM10 change acutely respectively, and the their maximum concentrations are 0.631 mg/m3 and 0.217 mg/m3 which exceed indoor air quality standard and the second class of ambient air quality standard. Polluting state of Chongqing toll is the worst among the four sampled tolls, and three indexes are bigger than others. Indoor and outdoor air pollutants have correlativity. Correlations of CO, PM10 and NO2 are significant at the 0.01 level respectively, and correlations between indoor and outdoor THC are significant at the 0.05 level. In the influencing factors analysis, traffic flow is significantly correlative with NO2, THC and PM10 (p < 0.01 or 0.01 < p < 0.05), and not significantly correlative with CO (p > 0.01). Air pressure and ambient temperature are predominating factors which influencing the concentration variation, and wind speed is a minor meteorological factor influencing the fluctuations of the data.