Concordance between reflectance confocal microscopy and histopathology for the diagnosis of acral lentiginous melanoma.

BACKGROUND: Acral lentiginous melanoma (ALM) is a highly malignant and invasive type of melanoma with unique locations of onset. Its incidence is increasing and early diagnosis is challenging. Reflectance confocal microscopy (RCM) is a non-invasive technique that provides an accurate image of tissue pathology. There are few reports on the use of RCM for the assessment of ALM. MATERIALS AND METHODS: In this retrospective study, data from 31 patients with a clinical diagnosis of ALM were collected. RCM image features were compared with histopathological findings to determine the concordance between the two methods. The sensitivity, specificity, positive predictive value, and negative predictive value of RCM for the diagnosis of ALM were evaluated. RESULTS: RCM and histopathology findings were concordant in 29 of 31 patients (93.5%). There were no false-negative results, although there were two false positives in RCM diagnosis. The sensitivity of RCM for diagnosing ALM was 100%, specificity was 50%, positive predictive value was 93.1%, and negative predictive value was 100%. CONCLUSIONS: RCM showed substantial concordance with histopathology in the diagnosis of ALM. It is a reliable and valuable non-invasive diagnostic tool that holds promise for the early diagnosis of ALM.

Mesenchymal stromal cell-derived magnetic nanovesicles for enhanced skin retention and hair follicle growth.

BACKGROUND AIMS: Extracellular vesicles and exosome-mimetic nanovesicles (NVs) derived from mesenchymal stromal cells (MSCs) have emerged as promising to promote hair growth. However, short local skin retention after subcutaneous administration hinders their clinical applications. METHODS: In this study, we prepared magnetic nanovesicles (MNVs) from iron oxide nanoparticle-incorporated MSCs. MNVs contained more therapeutic growth factors than NVs derived from naive MSCs, and their localization and internalization were manipulated by external magnetic field. RESULTS: Following the subcutaneous injection of MNVs into a mouse model of depilation-induced hair regeneration, the magnetic attraction increased their skin retention. Then, the cellular proliferation and ß-catenin signaling in hair follicles (HF) were markedly enhanced by MNV injection and magnetic field application. Furthermore, an acceleration of HF growth was revealed by histological analysis. CONCLUSIONS: The proposed strategy can enhance the therapeutic potential of MSC-derived NVs for hair regeneration and other dermatological diseases.

Reflectance Confocal Microscopy for Identification of Vulvar Lichen Sclerosus et Atrophicus and Vitiligo.

ABSTRACT: Lichen sclerosus et atrophicus (LSA) is a chronic inflammatory disease. The clinical presentation is clustered distribution of porcelain white spots that can be fused together. Early diagnosis of LSA is difficult, especially in the vulva, where it is difficult to distinguish from vitiligo. Traditional diagnostic methods mainly rely on histopathology. In this study, we used reflectance confocal microscopy (RCM) to study the characteristics of LSA and vitiligo. We investigated the clinical and confocal features of 40 cases of histopathologically proven genital LSA or vitiligo, and correlated the features with histopathologic findings. Through the analysis of the results, by RCM, we can easily distinguish genital LSA from vitiligo, which are clinically indistinguishable. This can reduce the number of unnecessary biopsies, and help with early diagnosis and timely treatment.

Neural progenitor cell-derived nanovesicles promote hair follicle growth via miR-100.

BACKGROUND: Accumulating evidence shows that mesenchymal stem cell-derived extracellular vesicles (EVs) hold great promise to promote hair growth. However, large-scale production of EVs is still a challenge. Recently, exosome-mimetic nanovesicles (NV) prepared by extruding cells have emerged as an alternative strategy for clinical-scale production. Here, ReNcell VM (ReN) cells, a neural progenitor cell line was serially extruded to produce NV. RESULTS: ReN-NV were found to promote dermal papilla cell (DPC) proliferation. In addition, in a mouse model of depilation-induced hair regeneration, ReN-NV were injected subcutaneously, resulting in an acceleration of hair follicle (HF) cycling transition at the site. The underlying mechanism was indicated to be the activation of Wnt/ß-catenin signaling pathway. Furthermore, miR-100 was revealed to be abundant in ReN-NV and significantly up-regulated in DPCs receiving ReN-NV treatment. miR-100 inhibition verified its important role in ReN-NV-induced ß-catenin signaling activation. CONCLUSION: These results provide an alternative agent to EVs and suggest a strategy for hair growth therapy.

Reflectance Confocal Microscopy Follow-up of Multifocal Superficial Basal Cell Carcinomas Treated With Imiquimod 5% Cream.

Introduction: Patients with multifocal superficial basal cell carcinomas (sBCC) require a non-invasive treatment and follow-up with a non-invasive technique. Imiquimod 5% cream is a new non-invasive therapy for BCC. Reflectance confocal microscopy (RCM) is a non-invasive, real-time imaging technique. Objectives: To evaluate and describe the feasibility and efficacy of imiquimod 5% cream for the treatment of multifocal sBCC using RCM. Methods: The efficacy of imiquimod 5% cream for the treatment of multifocal sBCC was evaluated, as well as the potential of RCM for assessing therapeutic effects. We reported four patients with 34 sBCC lesions were treated with imiquimod 5% cream. RCM was performed in the baseline and at 12 weeks, 24 weeks and 52 weeks after starting treatment. Results: Of 34 lesions treated with imiquimod 5%, 32 responded to the treatment and showed complete clinical clearing. Two subclinical BCC lesions were identified by RCM. The complete tumor clearance rate was 88.2%, and the efficiency rate was 97.1%. No lesion recurred at 24-month follow-up. RCM identified previously described confocal features of BCC and was more sensitive than clinical examination. Local skin reactions were relieved after expectant treatment. Conclusions: Imiquimod 5% cream may be useful for the treatment of multifocal sBCC, and its side effects are easy to manage. RCM can be used for non-invasive monitoring of treatment response and improved the tumor clearance rate.

CircMYC Regulates Glycolysis and Cell Proliferation in Melanoma.

Circular RNAs (cicRNAs) have been identified to play pivotal roles in several cancer types. However, functions of circRNA in malignant melanoma are poor defined. Our current study demonstrated that human circMYC was obviously upregulated in human melanoma tissue. Furthermore, circMYC promoted the proliferation of human melanoma cells and Mel-CV cells. The expression of circMYC can repress Mel-CV cell glycolysis and LDHA activities in the in vitro glycolysis and lactate production evaluations. circMYC directly bound to miR-1236 as a molecular sponge that targeting miR-1236 in Mel-CV cells via bioinformatics analysis, pull-down assay, and luciferase reporter assays. Our present study revealed that 3' UTR of LDHA acted as a target of miR-1236 using Mel-CV cells. Based on our findings, c-MYC-SRSF1 axis may regulate the production of circMYC. Overall, these results elucidate potential effects of circMYC in melanoma development and provide a promising biomarker for melanoma diagnosis.

Role of Annexin A2 isoform 2 on the aggregative growth of dermal papillae cells.

The dermal papilla is a major component of hair, which signals the follicular epithelial cells to prolong the hair growth process. Human Annexin A2 was preliminarily identified by two-dimensional gel electrophoresis (2-DE), MALDI-TOF-MS and database searching. The aim of the present study was to explore the role of Annexin A2 in the aggregative growth of dermal papillae cells (DPC). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were adopted to detect the expression of Annexin A2. And siRNA technique was used to suppress the expression of Annexin A2. Construction of over-expression vector was used to up-regulate the expression of Annexin A2. Cell Counting Kit 8 (CCK-8) and proliferating cell nuclear antigen (PCNA) were taken to detect the proliferation of DPC. The expression of Annexin A2 mRNA was up-regulated in passage 3 DPC compared with passage 10 DPC by RT-PCR. In line with the results at the mRNA level, Western blot analysis revealed that Annexin A2 isoform 2 was up-regulated significantly in passage 3 DPC compared with passage 10 DPC. The Annexin A2 isoform 2 siRNA was synthesized and transfected into passage 3 DPC. RT-PCR data showed the mRNA expression of Annexin A2 isoform 2 was suppressed in passage 3 DPC. Western blot results showed the expression level of Annexin A2 isoform 2 and PCNA were suppressed in passage 3 DPC. CCK-8 results showed that the proliferation of passage 3 DPC was suppressed (P < 0.05). Recombinant plasmid PLJM-Annexin A2 isoform 2-expression vector were constructed and were transfected into passage 10 DPC. RT-PCR data showed the mRNA expression of Annexin A2 isoform 2 was up-regulated in passage 10 DPC. Western blot results showed the expression level of annexin A2 isoform 2 and PCNA were up-regulated in passage 10 DPC. CCK-8 assay showed the proliferation of DPC was stimulated compared with control group (*P < 0.05). Our study proved that Annexin A2 isoform 2 may participate in regulating the proliferation of DPC and may be related to aggregative growth of dermal papilla cells. Therefore, our study suggests that Annexin A2 may be linked to hair follicle growth cycle.

Long-term effect of early-life supplementation with probiotics on preventing atopic dermatitis: A meta-analysis.

The prevalence of allergic diseases has increased over the past few decades. Atopic dermatitis (AD) is a common allergic disease, for which there is currently no known cure. Administration of probiotics in early life may be an effective method to prevent AD, but very little is known about its long-time preventive effect. In this research, a meta-analysis has been conducted to evaluate the long-term effect of early-life supplementation with probiotics on preventing AD. Meta-analysis was performed by the Review Manager version 5.2 software. Risk ratio and 95% confidence intervals were calculated by a fixed effect model. Six trials and a total of 1955 patients were included in this meta-analysis. The combined risk ratio of the meta-analysis comparing probiotics with placebo for investigating the long-term preventive effect of AD was 0.86 (95% CI 0.77-0.96), which demonstrates that probiotics is likely to produce long-term prevention of AD.

Low-Dose Topical 5-Aminolevulinic Acid Photodynamic Therapy in the Treatment of Different Severity of Acne Vulgaris.

The objective of this article is to investigate the effectiveness and safety of photodynamic therapy (PDT) with 3.6 % topical aminolevulinic acid (ALA) and a short incubation time with red light in moderate to severe acne. One hundred and thirty-six patients with moderate to severe acne were treated with 3.6 % topical ALA-PDT for three sessions with an interval of 2 weeks. Patients were evaluated for efficacy and safety on week 2, 4, 6, 8, and 12 after the initial treatment. Most patients showed apparent clearance of acne lesions at the treated site after three sessions. The effective treatment rates were increased after the multiple therapies. The clinical outcomes are the best at 4 weeks after the final treatment. The total effectiveness rate and cure rate of the low-dose ALA-PDT procedure is 92.65 and 47.06 %, respectively. Thirty-one patients and nineteen patients showed apparent exacerbation of acne lesions before the 2nd and 3rd treatment, respectively, but all of them showed good or excellent improvement after a three-course treatment. A few patients showed mild relapse including papules and comedos at 8 weeks after the final treatment. No significant differences are found in the effects of different acne severity and different genders. Adverse reactions are mild and transient. A 3.6 % topical ALA-PDT with a short time incubation with red light is a simple and an effective treatment option for moderate to severe acne with mild side effects in Chinese people.

[Construction and expression of scFv phage antibody from natural anti-keratin monoclonal antibody 3B4].

AIM: To construct and express the scFv phage antibody from natural anti-keratin monoclonal antibody 3B4 in E.coli and then compare its antigenic specificity with that of the parental antibody 3B4. METHODS: The V(H) and V(L) of 3B4 were amplified by PCR from pMD-18T- mAb 3B4 V(H) and pMD-18T- mAb 3B4 V(L), and then cloned into the phagemid pscMH. E.coli was transfected by recombined pscMH and superinfected by the helper phage VCSM13. Then the antigenic specificity of the scFv concentrated from the supernatant of the transfected E.coli was detected by indirect ELISA. RESULTS: DNA sequence analysis showed that the scFv sequence was the same as the original sequence except that there was one base mutation in the FR1 of V(L). Indirect ELISA showed that the scFv phage antibody was polyreactive as mAb 3B4, but its antigenic specificity was not the same with that of mAb 3B4. CONCLUSION: The scFv phage antibody from natural anti-keratin monoclonal antibody 3B4 was successfully constructed and expressed. Its antigenic specificity was different from that of its parental antibody 3B4.

[Reactivity of IgG class of anti-keratin autoantibodies in healthy human sera].

AIM: To analyze the reaction patterns of IgG class of anti-keratin autoantibodies (AK auto Abs) in normal human sera before and after purification, and to explore the effects of serum non-IgG components on the reaction patterns of IgG class of AK auto Abs. METHODS: Titers of IgG AK auto Abs in healthy human sera before and after purification were measured by indirect ELISA, and the reaction patterns of the AK auto Abs to a group of keratins with different relative molecule mass were analysed by Western blot. RESULTS: Titers of purified IgG class of AK auto Abs were higher than those of unpurified sera. Western blot analysis showed purified AK auto Abs recognized more keratins with stronger avidity than unpurified AK auto Abs. The reaction patterns of AK auto Abs from different individuals tended to be homogenous after purification. CONCLUSION: The reaction patterns of serum IgG class of AK auto Abs are influenced by some non-IgG components in sera. The homogeneity of reaction patterns of AK auto Abs from different individuals indicated that B cells producing IgG class of AK auto Abs may be selected by a set of conserved self-antigens, and this process may have no relationship with an individual's exposure to foreign antigens.

[Construction and expression of eukaryotic expression vector for intact IgG against human keratin].

AIM: To construct an eukaryotic expression vector containing the gene of anti-keratin antibody and express it in CHO(dhfr(-)) cells. METHODS: Both the V(L) and V(H) genes of an anti-keratin Fab from a phage display library were amplified by PCR. Using PCR product as the template, the V(kappa) and V(H) genes with the leader sequences (named L(kappa) and L(H) respectively) were amplified by overlapping PCR. After digested with endonuclease Xba I/BamH I and Xho I/Hind III, L(kappa) and L(H) genes were inserted into pWD digested with Xba I/BamH I and Xho I/Hind III, respectively to construct pWDkappaH. After PCR and sequencing, the expression plasmid pWDkappaH was transfected into CHO (dhfr(-)) cells. The culture supernatant of the transfected cells was collected and assayed for IgG activity. RESULTS: The eukaryotic expression vector pWDkappaH was constructed successfully and expressed in CHO(dhfr(-)) cells. The expression of intact IgG against keratin was identified by ELISA and RT-PCR. CONCLUSION: The successful expression of intact IgG against keratin lays the foundation for its clinical application.