Effect of black cloth ointment on hypertrophic scar formation: An investigation using integrated network pharmacology and animal assay.

BACKGROUND: Hypertrophic scars (HS) are a common disfiguring condition in daily clinical encounters which brings a lot of anxieties and concerns to patients, but the treatment options of HS are limited. Black cloth ointment (BCO), as a cosmetic ointment applicable to facial scars, has shown promising therapeutic effects for facial scarring. However, the molecular mechanisms underlying its therapeutic effects remain unclear. MATERIAL AND METHODS: Network pharmacology was first applied to analyze the major active components of BCO and the related signaling pathways. Subsequently, rabbit ear scar model was successfully established to determine the pharmacological effects of BCO and its active component ß-elemene on HS. Finally, the molecular mechanism of BCO and ß-elemene was analyzed by Western blot. RESULTS: Through the network pharmacology, it showed that ß-elemene was the main active ingredient of BCO, and it could significantly improve the pathological structure of HS and reduce collagen deposition. BCO and ß-elemene could increase the expression of ER stress-related markers and promote the increase of apoptotic proteins in the Western blot experiment and induce the apoptosis of myofibroblasts. CONCLUSIONS: Our findings indicate that the material basis for the scar-improving effects of the BCO is ß-elemene, and cellular apoptosis is the key mechanism through which the BCO and ß-elemene exert their effects.

Topical metformin suppresses angiogenesis pathways induced by pulsed dye laser irradiation in animal models.

Pulsed dye laser (PDL) is the first-line treatment for port-wine stain (PWS). However, only a small portion of the lesions could be completely cleared by PDL treatment, which might be related to the regeneration and revascularization of the vascular structures after laser irradiation. Recently, it is believed that the suppression of regeneration and revascularization of photocoagulated blood vessels can achieve a better therapeutic outcome. We use rabbit ear and SD rat as the animal models to investigate whether PDL-induced angiogenesis can be suppressed by topical metformin. Our results showed that topical application of metformin can effectively suppress the PDL-induced early stage of angiogenesis via inhibition of the AKT/mTOR/P70S6K pathway in animal models.

Novel GNE mutations in three Chinese patients with typical GNE myo-pathy.

GNE myopathy is an adult-onset muscle disorder featuring distal muscle atrophy and weakness. Rimmed vacuoles found in the muscle biopsies and gene mutations lead to the diagnosis of GNE myopathy. We collected clinical information, performed muscle biopsies and genetic testing on three patients. These cases developed typical disease presentations with distal muscle weakness at the ages of 26, 23, and 37 years. Their muscle pathologies revealed rimmed vacuoles. Genetic analysis led to the findings which included, c.1543-1544delGA (p.D515QfsX2)/c.38G>C (p.C13S) compound heterozygous mutation, c.733A>G (p.K245E) homozygous mutation and c.527A>T (p.D176V)/c.1634-1G>C (splicing) ; in which c.1543-1544 del GA (p.D515QfsX2), c.733A>G (p.K245E) and c.1634-1G>C (splicing) are three de novo mutations that have never been reported before. In conclusion, this study broadens the mutational spectrum of the GNE gene.

An association study of TOLL and CARD with leprosy susceptibility in Chinese population.

Previous genome-wide association studies (GWASs) identified multiple susceptibility loci that have highlighted the important role of TLR (Toll-like receptor) and CARD (caspase recruitment domain) genes in leprosy. A large three-stage candidate gene-based association study of 30 TLR and 47 CARD genes was performed in the leprosy samples of Chinese Han. Of 4363 SNPs investigated, eight SNPs showed suggestive association (P < 0.01) in our previously published GWAS datasets (Stage 1). Of the eight SNPs, rs2735591 and rs4889841 showed significant association (P < 0.001) in an independent series of 1504 cases and 1502 controls (Stage 2), but only rs2735591 (next to BCL10) showed significant association in the second independent series of 938 cases and 5827 controls (Stage 3). Rs2735591 showed consistent association across the three stages (P > 0.05 for heterogeneity test), significant association in the combined validation samples (Pcorrected = 5.54 × 10(-4) after correction for 4363 SNPs tested) and genome-wide significance in the whole GWAS and validation samples (P = 1.03 × 10(-9), OR = 1.24). In addition, we demonstrated the lower expression of BCL10 in leprosy lesions than normal skins and a significant gene connection between BCL10 and the eight previously identified leprosy loci that are associated with NFκB, a major regulator of downstream inflammatory responses, which provides further biological evidence for the association. We have discovered a novel susceptibility locus on 1p22, which implicates BCL10 as a new susceptibility gene for leprosy. Our finding highlights the important role of both innate and adaptive immune responses in leprosy.

Identification of IL18RAP/IL18R1 and IL12B as leprosy risk genes demonstrates shared pathogenesis between inflammation and infectious diseases.

Of eight leprosy susceptibility loci identified by genome-wide association studies, five have been implicated in Crohn disease, suggesting a common genetic fingerprint between leprosy and inflammatory bowel disease (IBD). Here, we conducted a multiple-stage genetic association study of 133 IBD susceptibility loci in multiple leprosy samples (totaling 4,971 leprosy cases and 5,503 controls) from a Chinese population and discovered two associations at rs2058660 on 2q12.1 (p = 4.57 × 10(-19); odds ratio [OR] = 1.30) and rs6871626 on 5q33.3 (p = 3.95 × 10(-18); OR = 0.75), implicating IL18RAP/IL18R1 and IL12B as susceptibility genes for leprosy. Our study reveals the important role of IL12/IL18-mediated transcriptional regulation of IFN-γ production in leprosy, and together with previous findings, it demonstrates the shared genetic susceptibility between infectious and inflammatory diseases.

Sustained activation of c-Jun N-terminal and extracellular signal-regulated kinases in port-wine stain blood vessels.

BACKGROUND: Port-wine stain (PWS) is a congenital, progressive vascular malformation but the pathogenesis remains incompletely understood. OBJECTIVE: We sought to investigate the activation status of various kinases, including extracellular signal-regulated kinase, c-Jun N-terminal kinase, AKT, phosphatidylinositol 3-kinase, P70 ribosomal S6 kinase, and phosphoinositide phospholipase C γ subunit, in PWS biopsy tissues. METHODS: Immunohistochemistry was performed on 19 skin biopsy samples from 11 patients with PWS. RESULTS: c-Jun N-terminal kinase, extracellular signal-regulated kinase, and P70 ribosomal S6 kinase in pediatric and adult PWS blood vessels were consecutively activated. Activation of AKT and phosphatidylinositol 3-kinase was found in many adult hypertrophic PWS blood vessels but not in infants. Phosphoinositide phospholipase C γ subunit showed strong activation in nodular PWS blood vessels. LIMITATION: Infantile PWS sample size was small. CONCLUSION: Our data suggest a subsequent activation profile of various kinases during different stages of PWS: (1) c-Jun N-terminal and extracellular signal-regulated kinases are firstly and consecutively activated in all PWS tissues, which may contribute to both the pathogenesis and progressive development of PWS; (2) AKT and phosphatidylinositol 3-kinase are subsequently activated, and are involved in the hypertrophic development of PWS blood vessels; and (3) phosphoinositide phospholipase C γ subunit is activated in the most advanced stage of PWS and may participate in nodular formation.

The differential diagnosis of lichenoid keratosis and seborrheic keratosis with reflectance confocal microscopy: a preliminary study.

BACKGROUNDS AND OBJECTIVES: Lichenoid keratosis (LK, or lichen planus-like keratosis, LPLK) and seborrheic keratosis (SK) present as similar benign keratotic lesions on cosmetically sensitive area, but require different therapies. Both lesions can be easily differentiated based on histological evaluation of biopsy materials. However, the biopsies may cause scarring and result in hyper-pigmentation, which reduces the compliance of the patients to be treated. In this study, we investigated the role of reflectance confocal microscopy (RCM) in the non-invasive differential diagnosis of LK and SK. PATIENTS AND METHODS: Cases with facial brown patches or plaques suspicious of SK were enrolled in the study. After written informed consent was obtained, the lesions were photographed, imaged by RCM, and then biopsied. The RCM findings were analyzed and correlated with histology results. Evaluation of the RCM pictures and confirmation with histological results were conducted by two independent dermatologists. RESULTS: In total, 10 cases were enrolled in the study. The main characteristics of LK lesions observed by RCM were the disarray of the dermal-epidermal junction (DEJ), and marked inflammatory infiltrates in the superficial dermis; while prominent cerebriform pattern, or elongated cords with bulbous projections without significant inflammation reaction, were the features of SK. Among the 10 cases, clinically suspicious of facial SK, 4 were determined as LK, 6 as SK by RCM imaging, and all the RCM findings were confirmed by histological results. CONCLUSIONS: The RCM features of LK and SK have significant difference, highlighting the important role of RCM in the differential diagnosis of LK and SK, avoiding biopsies and allowing safe treatments.

Genomewide association study of leprosy.

BACKGROUND: The narrow host range of Mycobacterium leprae and the fact that it is refractory to growth in culture has limited research on and the biologic understanding of leprosy. Host genetic factors are thought to influence susceptibility to infection as well as disease progression. METHODS: We performed a two-stage genomewide association study by genotyping 706 patients and 1225 controls using the Human610-Quad BeadChip (Illumina). We then tested three independent replication sets for an association between the presence of leprosy and 93 single-nucleotide polymorphisms (SNPs) that were most strongly associated with the disease in the genomewide association study. Together, these replication sets comprised 3254 patients and 5955 controls. We also carried out tests of heterogeneity of the associations (or lack thereof) between these 93 SNPs and disease, stratified according to clinical subtype (multibacillary vs. paucibacillary). RESULTS: We observed a significant association (P<1.00x10(-10)) between SNPs in the genes CCDC122, C13orf31, NOD2, TNFSF15, HLA-DR, and RIPK2 and a trend toward an association (P=5.10x10(-5)) with a SNP in LRRK2. The associations between the SNPs in C13orf31, LRRK2, NOD2, and RIPK2 and multibacillary leprosy were stronger than the associations between these SNPs and paucibacillary leprosy. CONCLUSIONS: Variants of genes in the NOD2-mediated signaling pathway (which regulates the innate immune response) are associated with susceptibility to infection with M. leprae.

Treatment of striae albae with 1,550 nm Er: Glass vs. CO2 fractional laser: A self-controlled study.

Objective: The objective of this study was to compare the efficacy and safety of fractional CO2 laser and 1,550 nm Er: glass laser in the treatment for the patients with striae albae. Methods: The female adults with striae albae in the abdomen for at least 12 months were recruited. After informed consent obtained, the patient received three treatments at 2-month intervals. The lesions on the left abdomen were treated with 10,600 nm CO2 fractional laser and right side treated with 1,550 nm Er: glass fractional laser. The pictures were taken before each visit and 3 months after the final treatment. The criteria for the evaluations using a quartile grading scale were excellent (76-100%), good (51-75%), fair (26-50%), poor (1-25%), and no improvement (0%). The safety and efficacy of the two lasers were independently evaluated using before and after photographs by two dermatologists. In addition, the self-reports to investigate the pain and satisfaction from patients were also recorded. Results: Totally, 27 cases were recruited, and 25 patients completed the treatments and follow-up. The excellent and good results (improvement of 51-100%) were achieved on the right abdomen in 84% of the patients, while 48% on the left site (p < 0.05). Hyper-pigmentation was seen in 20% of the patients assessed on the left abdomen and only in 8% on the right abdomen. During the treatments, average score of the pain on the right abdomen was 5.41 ± 2.13, which was higher than that on the left (4.19 ± 2.12) (p < 0.001). No permanent hyper-pigmentation was found on the both sides. Considering the whole treatments, the patients favored the modality used on the right side (80 vs. 68%, p < 0.05). Conclusion: Compared with CO2 fractional laser, 1,550 nm Er: glass fractional laser therapy provides the significantly better clinical outcomes and fewer side effects in the treatment of striae albae. Limitations: The sample size and follow-up time were limited.

Gene expression analysis of leprosy by using a multiplex branched DNA assay.

Leprosy is caused by Mycobacterium leprae, and the global registered prevalence of leprosy at the beginning of 2009 stood at 213,036 cases. It has long been thought that leprosy has a strong genetic risk. Recently, we have identified significant associations (P < 1.00 × 10(-10)) between SNPs in the genes CCDC122, C13orf31, NOD2, TNFSF15, HLA-DR and RIPK2 and a trend towards an association (P = 5.10 × 10(-5)) with a SNP in LRRK2. Here, we investigated the expression of these seven genes in formalin-fixed, paraffin-embedded skin tissues of leprosy and matched normal tissues using branched DNA technology. This technology allows for direct measurement of targeted mRNA within cellular lysate using a 96-well plate format in a time frame compared to a reporter gene assay. The clear upregulation of all seven genes was found in leprosy tissues compared to normal tissues, which further supports our genome-wide association study results.

Histological classification of melasma with reflectance confocal microscopy: a pilot study in Chinese patients.

OBJECTIVE: To investigate the histological classification of melasma with reflectance confocal microscopy (RCM) in vivo. METHODS: Two hundred and ten cases with facial melasma lesions were enrolled. After informed consent, the target melasma lesion of 10 patients were imaged with RCM and then biopsied as well. Under the RCM scanning, the distribution of the melanin determined the histological types, and then, the results of RCM images were compared with those of the histopathology. The other 200 cases were tested only with RCM. RESULTS: For the 10 cases imaged and biopsied, compared with that of the perilesional normal skin, the amount of melanin was significantly increased in the epidermis in all lesions under RCM, while three cases also found melanin in the dermis. Thus, seven of the 10 patients were categorized as the epidermal type while the other three as mixed ones, and the results were well correlated with those of the histopathology. Of the other 200 patients, 143 cases 71.5%) were categorized as the epidermal type while the other 57 (28.5%) cases as mixed ones. LIMITATIONS: If more melasma cases are biopsied, the data will be more convincing. CONCLUSION: RCM in vivo analysis shows complete coherence with histopathology results, which could be an alternative for the classification of melasma, and based on the results of RCM imaging, melasma is classified into two major types: the epidermal type and mixed type.

Imaging of Vulva Syringoma With Reflectance Confocal Microscopy.

Objectives: To investigate the application of reflectance confocal microscopy (RCM) imaging in diagnosis of vulva syringoma. Methods: Patients with lesions suspicious of syringoma on vulva were enrolled in the study. After informed consent was taken, the lesions were photographed and imaged with RCM. The features of the lesion in confocal images were then analyzed and compared with the biopsy findings for histology correlation. Results: Eleven cases in total were included in the study. The typical RCM features observed in syringoma are the presence of round to oval high refractive, and relatively monomorphous mass of varying sizes in the superficial and middle dermis, usually surrounded with 1-2 layers of light-dark line structures, which were further confirmed by histological evaluation. Ten cases showed classic features of syringoma and 1 case exhibited milia in RCM images. Conclusions: Syringoma has distinct features in RCM imaging, which correlates well with histological findings, highlighting the potential role of RCM in the diagnosis and differential diagnosis of vulva syringoma.

Seborrheic keratosis or verruca plana? A pilot study with confocal laser scanning microscopy.

BACKGROUND: Differentiation of some seborrheic keratosis (SK) and verruca plana (VP) lesions is a challenge. Confocal laser scanning microscopy (CLSM) has been proved to be useful in the diagnosis of skin diseases; however, to date, there is no report on the differential study of the two diseases with CLSM. OBJECTIVES: To obtain the CLSM image characteristics of SK and VP, and then test the differential ability of CLSM imaging. METHODS: We recruited 10 patients with typical lesions of SK under CLSM images to validate the features reported. Another 10 patients with typical VP lesions were also recruited, imaged with CLSM and biopsied to obtain the features under CLSM images based on histology analysis. Then, we attempt to summarize and refine those characteristics collected to obtain the most significant ones. All the cases with lesions suggestive of SK or VP were advised to undergo imaging with CLSM, and if CLSM imaging reflected discordantly with the clinical diagnosis, a biopsy was suggested for the exact lesion imaged. Those cases with CLSM and histology results were collected. Finally, two clinical dermatologists, who had no previous experience with CLSM, were tested with the simplified features of CLSM images to differentiate the suspected lesions of SK and VP among the cases collected. RESULTS: In total, there were 58 cases with CLSM images and histology results collected, in which, 40 cases were diagnosed as SK and 18 cases as VP by histology. The two blinded dermatologists' judgments were identical to histology analysis. CONCLUSION: CLSM proved to be valuable in the differential diagnosis of SK and VP. The simplified characteristics were easily understood and acceptable to those with no previous experience of CLSM.

The Differential Diagnosis of Hypopigmented Mycosis Fungoides and Vitiligo With Reflectance Confocal Microscopy: A Preliminary Study.

Objective: To investigate the role of reflectance confocal microscopy (RCM) in the differential diagnosis of hypopigmented mycosis fungoides (HMF) and vitiligo. Methods: Cases with persistent hypopigmented patches, suspicious of early stage vitiligo, or HMF were imaged with RCM. The melanin contents and inflammatory conditions of the epidermis and superficial dermis of the lesions were compared with the same layers of the adjacent skin, and then, the imaged lesions were biopsied and analyzed by histology. Results: 15 cases were enrolled in this study, and based on the RCM findings, there was just slight or moderate reduction of melanin but no melanin absence in the basal cell layer of HMF lesions. The finding of monomorphous weakly refractile, oval to round cells on the basis of vesicle-like dark space was clearly elucidated in the epidermis of the lesions by RCM, which indicates the Pautrier's microabscesses on histopathology. Among those 15 cases, 13 cases were identified as HMF, and the other two cases were vitiligo, based on RCM findings, which were confirmed by histology analysis. Conclusions: The RCM findings correlated well with histology results in the screening of HMF, which indicates the RCM is an important tool in the early detection and differential diagnosis of HMF.

[Observation on therapeutic effect of filiform fire needle assisted 308 nm excimer laser on vitiligo of different parts].

OBJECTIVE: To compared the therapeutic effect between filiform fire needle assisted 308 nm excimer laser and simple 308 nm excimer laser on vitiligo of different parts. METHODS: Target lesions of 134 patients were divided into an observation group and a control group according to the principle of self-controlled, 201 pieces in each one. In the observation group, filiform fire needle was performed at target lesions. Then target lesions both of the two groups were irradiated with 308 nm excimer laser at the same time. Once every 2 weeks, totally 10 treatments were required. The effective rate and effective rate, color recovery rate and responding time of different parts in the two groups were evaluated 2 weeks after treatment. RESULTS: The effective rate in the observation group was 82.59% (166/201), which was higher than 68.16% (137/201) in the control group (P<0.01). The effective rate of face-neck, trunk, limbs and hand-foot were 90.32%, 81.63%, 81.48% and 58.62% respectively in the observation group, which were higher than 82.80%, 69.39%, 51.85% and 31.03% in the control group (P<0.01, P<0.05). The color recovery rate of different parts in the observation group was higher than the control group, and the effect was faster in the observation group (P<0.01, P<0.05). CONCLUSION: Filiform fire needle as an adjunctive therapy, combined with 308 nm excimer laser are more effective than simple 308 nm excimer laser for vitiligo of different parts. Combination therapy has a shorter responding time, the face-neck has the best effect and hand-foot has poor effect.

Identification of Seven Novel Mutations in the Acid Alpha-glucosidase Gene in Five Chinese Patients with Late-onset Pompe Disease.

BACKGROUND: Pompe disease is a rare lysosomal glycogen storage disorder linked to the acid alpha-glucosidase gene (GAA). A wide clinical and genetic variability exists between patients from different ethnic populations, and the genotype-phenotype correlations are still not well understood. The aim of this study was to report the clinicopathological and genetic characteristics of five Chinese patients with late-onset Pompe disease (LOPD) who carried novel GAA gene mutations. METHODS: Clinical and pathological data of patients diagnosed with glycogen storage disease at our institution from April 1986 to August 2017 were collected, and next-generation sequencing of frozen muscle specimens was conducted. RESULTS: Of the five patients included in the study, the median disease onset age was 13 years, with a median 5 years delay in diagnosis. The patients mainly manifested as progressive weakness in the proximal and axial muscles, while one patient developed respiratory insufficiency that required artificial ventilation. In muscle biopsies, vacuoles with variable sizes and shapes appeared inside muscle fibers, and they stained positive for both periodic acid-Schiff and acid phosphatase staining. Ten GAA gene mutations, including seven novel ones (c.796C>A, c.1057C>T, c.1201C>A, c.1780C>T, c.1799G>C, c.2051C>A, c.2235dupG), were identified by genetic tests. CONCLUSIONS: The seven novel GAA gene mutations revealed in this study broaden the genetic spectrum of LOPD and highlight the genetic heterogeneity in Chinese LOPD patients.

Pathological findings in sporadic inclusion body myositis and GNE myopathy.

OBJECTIVE: The following study compared the pathological findings between sporadic inclusion body myositis (sIBM) and Glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase myopathy (GNEM) patients. METHODS: An enzyme histochemistry was used to compare the pathological characteristics between 11 patients with sIBM and 16 patients with GNEM. RESULTS: There were four pathological differences observed: (1) A majority of the rimmed vacuoles found in the sIBM patients resembled cracks, whereas the GNEM patients (P=0.004) had round or oval vacuoles. (2) A majority of the rimmed vacuoles that were located in the periphery of the atrophic muscle fibers of the sIBM patients. The patients with GNEM had a majority of the rimmed vacuoles in the center of the atrophic muscle fibers (P=0.001). (3) The patients with sIBM had basophilic granules in the rimmed vacuoles, which appeared to be fine granules that were sand-like particles. The GNEM patients had coarse granules (P=0.018). (4) The proportion of mononuclear cells invasion of muscle fibers was larger in the sIBM patients than the GNEM patients (P=0.047). The GNEM patients were younger on average than the sIBM patients at the onset of symptoms (P<0.001) and at the diagnosis age (P<0.001). The electromyography (EMG) showed the presence of myogenic lesions in 10 patients with sIBM, both myogenic and neurogenic lesions in one patients with sIBM and myogenic lesions in 16 patients with GNEM. CONCLUSION: There were significant differences in the morphologies of the rimmed vacuoles between sIBM patients and GNEM patients.