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Objective:To investigate clinicopathological features of hypopigmented mycosis fungoides (HMF) and hypopigmented interface T-cell dyscrasia (HITCD) .Methods:A total of 41 patients with cutaneous hypopigmented lymphoproliferative diseases, who had complete clinicopathological data, were collected from Department of Dermatology, the Third People′s Hospital of Hangzhou from January 2015 to September 2020, and the clinicopathological and immunophenotypic features were analyzed. Comparisons of normally distributed measurement data were carried out using t test, comparisons of categorical data using Chi-square test or Fisher′s exact test, and comparisons of ranked data between 2 groups using rank-sum test. Results:All of the 41 patients clinically presented with irregular hypopigmentation, some of which was accompanied by erythema or furfuraceous scales. In terms of pathological features, 21 patients showed infiltration and aggregation of atypical lymphoid cells in the epidermis, which was consistent with typical pathological features of mycosis fungoides, and they were diagnosed with HMF; 20 patients showed vacuolar degeneration of the basal layer, accompanied by infiltration of lymphoid cells and mild epidermotropism, and they were diagnosed with HITCD. All immune cells expressed T-cell phenotype, and epidermal lymphocytes expressed a CD8-dominated phenotype in 14 (67%) cases of HMF and 13 (65%) of HITCD. In the epidermis, the total number of lymphocytes was significantly higher in the HMF group than in the HITCD group ( t= 1.81, P= 0.012) ; in the dermis, the number of CD4 + lymphocytes and CD8 + lymphocytes, and the total number of lymphocytes were all significantly higher in the HMF group than in the HITCD group ( t= 2.64, 1.51, 2.60, P= 0.012, 0.002, 0.001, respectively) . All patients were treated with narrow-band ultraviolet B radiation. Among 34 patients who completed the follow-up, 30 achieved complete clearance of skin lesions without recurrence, including all patients with HITCD, and 4 with HMF achieved partial regression of the lesions. Conclusions:Compared with HMF, HITCD presents different pathological characteristics and benign biological behaviors. Thus, HITCD should be distinguished from HMF as an independent disease. Phototherapy alone is effective for the treatment of HITCD.
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Objective:To assess the value of culture of epidermal melanocytes from negative-pressure suction blisters in the auxiliary diagnosis of segmental vitiligo-like nevus depigmentosus.Methods:Between June 2019 and March 2020, 8 patients with segmental vitiligo-like nevus depigmentosus, who met the Coupe′s clinical diagnostic criteria, were enrolled from Department of Dermatology, Hangzhou Third People′s Hospital. All patients were evaluated by the Wood′s lamp, reflectance confocal microscopy (RCM) , 308-nm excimer laser radiation, and in vitro culture of epidermal melanocytes from negative-pressure suction blisters. Results:Among the 8 patients, fluorescence was observed in 6 under the Wood′s lamp, dermal papillary rings were incomplete or absent in 4 as shown by RCM, and 5 experienced no repigmentation after 308-nm excimer laser radiation. Among the 8 patients, in vitro cultured lesional melanocytes were all positive for ferrous sulfate staining, yellowish-white precipitates were obtained after digestion and centrifugation of the melanocytes, and stage Ⅰ-Ⅲ melanosomes were observed in the cytoplasm of melanocytes under the electron microscope; however, the precipitates were black in color after digestion and centrifugation of the melanocytes collected from the normal skin tissues at the contralateral anatomical site, and stageⅠ-Ⅳ melanosomes were seen in the cytoplasm of the melanocytes under the electron microscope. Conclusion:Culture of epidermal melanocytes from negative-pressure suction blisters may facilitate the diagnosis of segmental vitiligo-like nevus depigmentosus.
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Objective:To investigate the efficacy of systemic glucocorticoid treatment and its related factors in progressive vitiligo patients with vitiligo disease activity (VIDA) scores ≥ 2 points.Methods:A total of 272 progressive vitiligo patients with VIDA scores ≥ 2 points and skin lesion area < 1% of body surface area, who received no systemic glucocorticoid treatment, were collected from Department of Dermatology, the Third People′s Hospital of Hangzhou from June 2018 to June 2019. The area and type of skin lesions, VIDA scores, predisposing factors and special clinical markers (trichrome vitiligo, confetti-like depigmentation, Koebner phenomenon and inflammatory vitiligo) were analyzed. These patients were randomly divided into 3 groups by a random number table: topical glucocorticoid group (62 cases) , oral prednisone + topical glucocorticoid group (76 cases) and compound betamethasone injection + topical glucocorticoid group (134 cases) , and the latter two groups were also called as the systemic and topical glucocorticoid group. The patients in the topical glucocorticoid group were treated with halometasone cream or 0.05% clobetasol propionate cream once a day; during the oral prednisone treatment, the dose was adjusted once every 7 days, and gradually reduced from 30 mg/d to 20, 15, 10 and 5 mg/d, and the treatment lasted 35 days; during the treatment with compound betamethasone injection, intramuscular injection was performed once every 20 days at a dose of 1 ml for 2 sessions. The stable disease rate (defined as the proportion of patients experiencing no progression during the study among the analyzed patients) was calculated in these groups after 3 months of treatment, and changes in vitiligo types were evaluated after 1 year of follow-up. Statistical analysis was carried out by using Kruskal-Wallis H test, χ2 test and Fisher′s exact test. Results:After 3-month treatment, there was a significant difference in the expansion rate of skin lesion area among the 3 groups ( H = 12.468, P < 0.001) , and the expansion rate of skin lesion area was significantly lower in the oral prednisone + topical glucocorticoid group and compound betamethasone injection + topical glucocorticoid group than in the topical glucocorticoid group ( P < 0.001, = 0.005, respectively, α = 0.016 7) ; among the patients with slowly progressive vitiligo (VIDA scores = 2 or 3 points) , the stable disease rate was significantly higher in the systemic and topical glucocorticoid group than in the topical glucocorticoid group ( χ2 = 23.973, 11.877, respectively, both P < 0.001) ; the stable disease rate also significantly differed among the patients with different VIDA scores (VIDA scores = 2, 3 or 4 points) in the systemic and topical glucocorticoid group ( χ2 = 17.122, P < 0.001) . After 3-month treatment, the patients with predisposing factors or special clinical markers showed significantly decreased stable disease rate (47.3% [35/74], 41.2% [47/114], respectively) compared with those without predisposing factors or special clinical markers (70.6% [96/136], 87.5% [84/96]; χ2 = 11.098, 47.548, respectively, both P < 0.001) . After 1 year of follow-up, the proportion of patients with localized vitiligo converted into non-localized vitiligo was significantly higher in the topical glucocorticoid group (41.9%, 26/62) than in the systemic and topical glucocorticoid group (21.9%, 46/210; χ2 = 10.328, P = 0.006) , and higher in the group with predisposing factors or special clinical markers than in that without predisposing factors or special clinical markers respectively (both P < 0.01) . Conclusions:Early systemic glucocorticoid treatment should be performed in the progressive vitiligo patients with high VIDA scores, predisposing factors and special clinical markers.
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Objective:To observe the efficacy of anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin in the treatment of severe acne.Methods:Fifty patients with severe acne were selected in the Dermatology Clinic of the Third People＇s Hospital of Hangzhou from November 2018 to July 2019. They were randomly divided into the experimental group of 25 cases and the control group of 25 cases. The experimental group was treated with anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin orally. The control group was treated with isotretinoin orally alone. Before and after treatment for 56 days, lactate score, skin cuticle hydration (SCH), transepidermal water loss (TEWL) and skin physiological indexes were measured.Results:After 56 days of treatment, the TEWL and SCH of the control group were 15.75±3.31 and 10.13±3.62, the TEWL and SCH of the experimental group were 12.17±3.61 and 28.07±3.17, respectively; the difference was statistically significant ( T was 3.610 and 12.398, P was 0.002 and 0.000, respectively). The volume and depth of cyst nodule, scar depression, skin roughness, absolute value and area of erythema in the experimental group were significantly lower than those in the control group ( T was 2.280, 1.676, 2.332, 1.508, 4.813 and 3.637; P was 0.031, 0.011, 0.027, 0.040, 0.000 and 0.001, respectively). Conclusions:Anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin has a good effect on severe acne and it can reduce the barrier damage and other adverse reactions.
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Objective:To investigate the role of folliculin in apoptosis of and chemokine secretion by melanocytes mediated by interferon-γ (IFN-γ) .Methods:Normal primary melanocytes were isolated from circumcised foreskin tissues from a healthy male child, and primary vitiliginous melanocytes were isolated from normally pigmented suction-blistered epidermis from patients with vitiligo after suction blister epidermal grafting. Western blot analysis was performed to determine the folliculin protein expression in normal primary melanocytes, primary vitiliginous melanocytes and a human primary melanocyte line PIG1. PIG1 cells stimulated with 10 ng/ml IFN-γ for 48 hours served as induction group, and untreated PIG1 cells served as control group. Real-time quantitative RCR (qRT-PCR) was performed to determine the mRNA expression of folliculin, autophagy-related microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ and Beclin genes, and Western blot analysis to determine the protein expression of folliculin, Beclin1 and LC3Ⅱ/Ⅰ, as well as phosphorylation levels of adenosine monophosphate-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) in the above cells. Furthermore, the melanocytes stimulated with 10 ng/ml IFN-γ for 48 hours were divided into several groups: negative control group infected with an empty lentiviral vector, folliculin inhibition group infected with a folliculin-inhibiting lentivirus, autophagy enhancement group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with a mTOR inhibitor, autophagy inhibition group infected with a folliculin-inhibiting lentivirus followed by 2-hour treatment with an AMPK inhibitor. Then, flow cytometry was conducted to detect apoptosis of PIG1 cells, and enzyme-linked immunosorbent assay to measure the concentration of chemokines CXCL10 and CCL20 in the culture supernatant of PIG1 cells in the above groups. Measurement data were compared among multiple groups by using one-way analysis of variance, and multiple comparisons were carried out by using least significant difference- t test. Results:The relative protein expression level of folliculin significantly differed among the normal primary melanocytes (0.850 ± 0.120) , primary vitiliginous melanocytes (1.507 ± 0.170) and PIG1 cells (0.697 ± 0.130; F = 50.09, P < 0.001) , and was significantly higher in the primary vitiliginous melanocytes than in the normal primary melanocytes and PIG1 cells ( t = 4.06, 5.89, respectively, both P < 0.01) . Compared with the control group, the induction group showed significantly increased relative mRNA and protein expression levels of folliculin (both P < 0.01) , but significantly decreased relative mRNA and protein expression levels of LC3Ⅱ and Beclin (all P < 0.01) ; moreover, the induction group showed significantly decreased LC3Ⅱ/Ⅰ levels (0.72 ± 0.02) and AMPK phosphorylation levels (0.714 ± 0.023) in the PIG1 cells compared with the control group (1.13 ± 0.02, 1.176 ± 0.002, t = 7.34, 6.67, respectively, both P < 0.01) , but significantly increased mTOR phosphorylation levels (1.051 ± 0.023) compared with the control Group (0.451 ± 0.016, t = 3.81, P = 0.009) . There were significant differences in the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 among the control group, induction group and other treatment groups (all P < 0.001) ; specifically, the PIG1 cell apoptosis rate and concentrations of CXCL10 and CCL20 were significantly higher in the induction group than in the control group, lower in the folliculin inhibition group than in the negative control group, lower in the autophagy enhancement group than in the folliculin inhibition group, and higher in the autophagy inhibition group than in the folliculin inhibition group (all P < 0.05) . Conclusions:Folliculin is highly expressed in vitiliginous melanocytes. Folliculin expression and downstream signaling pathways are regulated by IFN-γ, and folliculin may participate in IFN-γ-mediated melanocyte apoptosis and chemokine secretion via regulating autophagy.
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Objective:To investigate clinical efficacy and safety of cultured autologous melanocyte transplantation for the treatment of non-segmental vitiligo accompanied by autoimmune thyroid diseases.Methods:From May 2008 to December 2018, a total of 2 284 patients with non-segmental vitiligo were retrospectively collected, who received cultured autologous melanocyte transplantation in Hangzhou Third People′s Hospital. Among these patients, 75 were also diagnosed with autoimmune thyroid diseases, including hyperthyroidism (42 cases) , hypothyroidism (18 cases) and Hashimoto′s thyroiditis (15 cases) . Efficacy and safety were compared between the vitiligo patients with autoimmune thyroid diseases (concomitant group) and those without (non-concomitant group) . Chi-square test was used to compare enumeration data.Results:Among the 2 284 patients, 1 085 were males and 1 199 were females, with an age of 25.0 ± 1.2 years and a disease duration of 5.1 ± 2.3 years. Six months after transplantation, 1 873 out of 2 209 patients in the non-concomitant group achieved favorable clinical response, with a response rate of 84.8%, including 1 162 achieving complete clinical response (52.6%) ; 46 out of 75 patients in the concomitant group achieved favorable clinical response, with a response rate of 61.3%, including 20 achieving complete clinical response (26.7%) ; the response rate and recovery rate were both significantly lower in the concomitant group than in the non-concomitant group ( χ2 = 29.72, 19.54, respectively, both P < 0.001) . Moreover, the response rate was significantly lower in the hypothyroidism group than in the hyperthyroidism group ( χ2 = 6.61, P = 0.010) . The incidence of isomorphic response at the donor site was significantly higher in the concomitant group than in the non-concomitant group (9.3% vs. 4.3%, χ2 = 4.31, P = 0.038) , so were the recurrence rates of vitiliginous patches at the recipient site after 1, 3, 5 and 10 years (concomitant group: 6.7%, 14.7%, 17.3%, 8.7%, respectively; non-concomitant group: 0.7%, 1.4%, 2.1%, 3.6%, respectively; χ2 = 29.96, 70.69, 67.23, 41.61, respectively, all P < 0.001) . Conclusion:Concomitant autoimmune thyroid diseases negatively affect the efficacy of cultured autologous melanocyte transplantation in the treatment of vitiligo, so effective measures should be taken to prevent isomorphic response and recurrence at the recipient site for patients with non-segmental vitiligo complicated by autoimmune thyroid diseases.
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Objective:To investigate clinical efficacy of composite subcutaneous pedicled flaps in repairing full-thickness nasal alar defects.Methods:Clinical data were collected from 11 patients with basal cell carcinoma of nasal ala in Department of Dermatologic Surgery, the Third People′s Hospital of Hangzhou from August 2019 to September 2020, and analyzed retrospectively. There were 6 females and 5 males, aged 70 years in average (range, 56-87 years) . After tumor resection, full-thickness nasal alar defects were formed, a flap was designed along the inner side of the cheek and turned by 180 degrees to repair the nasal mucosal lining; then, a subcutaneous pedicled flap was designed along the ipsilateral nasolabial fold to repair the outer secondary defects and postoperative defects.Results:The mean diameter of postoperative defects was 1.8 cm (range, 1.5-2.5 cm) , and the mean diameter of the nasal lining was 0.9 cm (range, 0.6-1.5 cm) . Seven days after the operation, stitches were removed, wounds healed without complications, and all flaps survived with a satisfactory appearance. The follow-up period ranged from 1 to 13 months (average, 6 months). After scar maturation, all patients were satisfied with the aesthetic outcome. No tumor recurrence was observed during the follow-up period.Conclusion:The composite subcutaneous pedicled flap is safe and effective for the repair of full-thickness nasal alar defects, and has advantages of a well-hidden donor site, good color match, and stage-I operation for reconstruction.
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Objective@#To evaluate the efficacy of long-term treatment with tacrolimus ointment in patients with vitiligo.@*Methods@#A total of 156 patients with vitiligo (70 males and 86 females) were enrolled into this study, who were firstly diagnosed with vitiligo and continuously followed up in Department of Dermatology, Yiwu Dermatology Hospital and the Third People′s Hospital of Hangzhou between January 2016 and February 2018. Of the 156 patients, 114 were adults, and 42 were children aged 6 - 18 years. All the patients received 6-month treatment with tacrolimus ointment twice a day. They were followed up once a month, the time to initial repigmentation and vitiligo area severity index (VASI) were recorded, and VASI improvement rate was calculated. After 6-month treatment, the patients achieving marked improvement were divided into conventional treatment group and interval treatment group to be treated with tacrolimus ointment twice a day and once every 3 days respectively for another 6 months, and final efficacy of the 2 protocols was compared. Mauchly′s test of sphericity, randomized block analysis of variance and Bonferroni method were used to analyze differences among pre-treatment VASI scores, and 1- to 6-month post-treatment VASI scores.@*Results@#Among the 156 patients, the pre-treatment, and 1-, 2-, 3-, 4-, 5- and 6-month post-treatment VASI scores[M (P25, P75) ] were 2 (0.6, 5) , 2 (2, 5) , 1.6 (0.575, 4.5) , 1.2 (0.5, 4.0) , 0.4 (1.15, 3.5) , 0.75 (0.3, 2.575) , 0.6 (0.2, 2.2) respectively. Additionally, there was a significant difference in the VASI score among the above time points (F = 6.14, P < 0.05) , and the VASI score showed a gradual decreasing trend over time. During the initial 6-month treatment, the average time to initial repigmentation was 2.2 ± 0.6 months. Of the 156 patients, 98 (62.8%) received marked improvement, and the time to marked improvement was 5.2 ± 0.6 months. Of the 98 patients who received marked improvement, 46 completed another 6-month treatment. After 12-month treatment, there was no significant difference in the repigmentation rate between the conventional treatment group (73.3% ± 18.2%) and interval treatment group (72.0% ± 16.2%, t = 0.42, P > 0.05) .@*Conclusions@#The initial 6-month topical tacrolimus therapy showed marked efficacy for vitiligo. After the marked improvement of skin lesions, good efficacy can be maintained by long-term treatment with topical tacrolimus once every 3 days or twice a day.
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The pathogenesis of vitiligo is complicated, and the loss of melanocytes plays a central role. Besides deficiency in melanocytes, it is considered that dysfunction of epidermal and dermal cell populations as well as their interaction with melanocytes also play important roles in the occurrence of vitiligo, Thus, it is necessary for understanding and treatment of vitiligo to fully and accurately understand pathophysiological states of full-thickness vitiliginous skin cells and tissues at different stages. This review aims to summarize research progress in the role of melanocytes and related cell populations in the occurrence of vitiligo.
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Objective:To preliminarily evaluate the effect of Fam114A1 on the biological function of melanocytes.Methods:A375 human melanoma cells was used to construct stably Fam114A1-overexpressing or -inhibited cell line by lentiviral transfection, namely overexpression group and expression inhibition group respectively, and A375 cells transfected with an empty lentivirus served as control group. Real-time fluorescence-based quantitative PCR was performed to evaluate effect of Fam114A1 on the mRNA expression of melanin synthesis-related genes tyrosinase (TYR) , tyrosinase-related protein 1 (TYRP1) , premelanosome protein (PMEL) , microphthalmia-associated transcription factor (MITF) and dopachrome isomerase (DCT) , Western blot analysis was conducted to determine the protein expression of TYR and MITF, methyl thiazol tetrazolium (MTT) assay, Transwell migration and adhesion assays were conducted to assess the effect of Fam114A1 on cellular proliferative activity, migratory and adhesive ability of A375 cells respectively. Statistical analysis was carried out by using one-way analysis of variance and Dunnett- t test. Results:Fluorescence microscopy showed that lentivirus-based transfection efficiency was about 90% in the 3 groups. Compared with the control group (0.850 ± 0.120) , the protein expression of Fam114A1 significantly increased in the overexpression group (1.507 ± 0.170, t = 5.888, P = 0.001) , but significantly decreased in the expression inhibition group (0.397 ± 0.120, t = 4.065, P = 0.007) , suggesting that the stably Fam114A1-overexpressing or -inhibited A375 cell line was successfully constructed. Real-time fluorescence-based quantitative PCR and Western blot analysis showed that the mRNA and protein expression of TYR and MITF were significantly lower in the expression inhibition group than in the control group (all P < 0.01) , but did not differ between the overexpression group and control group (all P > 0.05) . Compared with the control group, the expression inhibition group showed significantly increased cellular proliferative activity and adhesive ability ( P = 0.009, 0.001, respectively) , but significantly decreased migratory ability ( P = 0.005) , while the overexpression group only showed significantly increased migratory ability ( P = 0.021) . Conclusions:Fam114A1 can affect the proliferative activity, migratory and adhesive abilities of A375 cells, and the expression of melanin synthesis-related proteins TYR and MITF in A375 cells. Fam114A1 may be a functional protein involved in regulating the biological activity of melanocytes.
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Objective:To analyze consistency between histopathological and reflectance confocal microscopy (RCM) characteristics of early-stage mycosis fungoides (MF) , and to evaluate the value of RCM in assisting the pathological diagnosis of early-stage MF and the feasibility of dynamic monitoring of treatment response with RCM.Methods:From January 2014 to January 2018, 40 cases of clinically suspected MF were collected from Department of Dermatology, Third People′s Hospital of Hangzhou, including 26 males and 14 females, and their age was 47.0 ± 17.6 years. According to the summarized RCM characteristics of early-stage MF, biopsy sites were preliminarily located, and then a histopathological examination was performed. The RCM and pathological features of MF were compared. In addition, a combination therapy with narrowband ultraviolet B and interferon was performed in patients with confirmed MF. Targeted lesions were followed up with RCM for 9 months, and then therapeutic efficacy was evaluated.Results:Among the 40 cases of clinically suspected MF, 8 were preliminarily diagnosed as typical MF, 18 as suspected MF, and 14 were excluded according to the RCM characteristics; according to the pathological features, 12 could be diagnosed as typical MF, 14 as suspected MF, and 14 were excluded. Consistency analysis showed that the kappa coefficient between RCM classification and pathological diagnosis was 0.848 ( P < 0.01) . The consistency of epidermal infiltration of mildly refractive cells was the highest between RCM and pathological findings (kappa coefficient = 1, P = 0.005) , followed by dermal fibrosis at the erythema stage (kappa coefficient = 0.714, P = 0.035) . The RCM characteristics of MF gradually returned to normal during treatment, but atypical lymphocytes still existed when clinical lesions completely regressed. Conclusion:RCM can be used for pathological localization of suspected MF lesions in the early stage, and for dynamic monitoring of therapeutic efficacy in MF.
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Objective:To evaluate the efficacy of sectioned hair follicle transplantation in the treatment of stable vitiligo in hair-bearing areas.Methods:A total of 10 patients with stable vitiligo aged 19-28 years (including 4 males and 6 females) were enrolled from Department of Dermatology, The Third People′s Hospital of Hangzhou between December 2016 and December 2018. Among the 10 patients, a total of 26 white patches accompanied by leukotrichia received sectioned hair follicle transplantation, including 18 on the scalp and 8 on the eyebrow. Hair follicles were obtained from the occipital and postauricular regions by follicular unit extraction, and divided into the upper and lower portions. Each white patch with leukotrichia was divided into 2 regions, and the 2 portions of the hair follicle were transplanted into the 2 regions of lesions respectively. The patients were followed up once every 4 weeks after grafting, and repigmentation and hair growth were evaluated.Results:At 12 weeks, of the 26 white patches transplanted with the upper hair follicles, 3 were cured, 5 were markedly improved, 12 were improved, and 6 showed no response; the response rate was 30.8% (8/26) , and no hair grew on the transplantation sites. Of the 26 lesions transplanted with the lower hair follicles, 5 were cured, 7 were markedly improved, 11 were improved, and 3 showed no response; the response rate was 46.2% (12/26) , and hair growth was observed on 8 lesions. There was no significant difference in the response rate between the upper hair follicle transplantation group and lower hair follicle transplantation group ( χ2 = 1.30, P = 0.25) . Conclusion:Sectioned hair follicle transplantation is suitable for the treatment of vitiligo complicated by leukotrichia on the scalp and eyebrow.
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Objective:To analyze clinical and pathological characteristics of 9 cases of hypopigmented mycosis fungoides (HMF) .Methods:Nine patients with clinically and histopathologically confirmed HMF were collected at Department of Dermatology, Hangzhou Third People's Hospital from 2014 to 2018, and their clinical and pathological manifestations and follow-up findings were retrospectively analyzed.Results:Among the 9 patients, there were 7 males and 2 females; the age ranged from 6 to 29 years (average, 12.4 years) ; the duration of disease varied from 6 months to 3 years (average, 20.7 months) . All skin lesions manifested as poorly demarcated, irregular, hypopigmented patches, some hypopigmented patches were confluent, and hypopigmented patches were covered with scales in 2 cases. Wood′s lamp examination of hypopigmented patches showed a grayish-white color with weak fluorescence. Dermoscopic images showed white scales on the lesions and blood vessels distributed in spermatozoa-like, punctiform or short linear patterns. Reflectance confocal microscopy showed mild hypopigmentation in the basal layer without obvious depigmentation, and highly-refractive atypical cells at the dermo-epidermal junction. Histopathological examination of the 9 cases showed epidermotropic phenomenon of atypical lymphocytes, and some cells clustered and formed Pautrier′s microabscess, immunohistochemical staining revealed that all the cells expressed T cell phenotypes, and were mainly infiltrating CD8 + T cells. The 9 patients were treated with narrow-band ultraviolet B (NB-UVB) , and all of them achieved clinical complete response. During a follow-up of 23.8 months on average, 2 patients experienced recurrence, and they achieved clinical complete response again after NB-UVB treatment. Conclusions:HMF progresses slowly, and NB-UVB treatment is effective. There is the possibility of recurrence after clinical recovery in patients with HMF, and a long-term follow-up is needed.
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Objective:To identify and differentiate cell subsets in the epidermis and dermis of vitiligo skin lesions using single-cell RNA sequencing technology, and to study the relationship between them.Methods:Skin samples were collected from 2 healthy people without immune or systemic diseases and 2 patients with stable non-segmental vitiligo in Department of Dermatology in the Third People′s Hospital of Hangzhou in September 2019. Single-cell transcriptome sequencing was performed on 11 000 cells in all the skin samples by using 10 × Genomics single-cell RNA-Seq technology. Cell subsets were analyzed, screened and counted by using Seurat software.Results:Cluster analysis of gene expression in the 2 normal skin tissues revealed several cell subsets, including keratinocytes, fibroblasts, nerve cells and melanocytes, endothelial cells, tissue stem cells, and immune cells mainly consisting of dendritic cells and T cells. In the 2 vitiligo lesions, abnormal differentiation and quantity were observed in fibroblasts and 4 keratinocyte subpopulations. The proportion of fibroblasts was significantly lower in vitiligo lesions than in normal skin tissues (0 vs. 0.4%) , while the proportions of keratinocyte subpopulations 5, 6, 10 and 12 (8.03%, 7.36%, 3.52%, 0.91%, respectively) in vitiligo lesions were significantly higher than those in the normal skin tissues (4.47%, 3.53%, 2.69%, 0.28%, respectively, all P < 0.01) . Moreover, the above keratinocyte subpopulations were at the end of cell differentiation, and expressed very significant and specific marker genes, which were mainly closely related to cell-cell interactions and cell homeostasis. GO and KEGG analysis showed that keratinocyte subpopulations 5 and 6 were mainly related to intercellular connection, cell adhesion and cytoskeleton function, while the keratinocyte subpopulation 10 was closely related to cell homeostasis. Conclusion:The single-cell sequencing technology was firstly used to study the transcriptional expression profile of vitiligo lesions in China, and preliminary analysis revealed 4 groups of keratinocytes with different quantity and functions, suggesting that abnormal differentiation and dysfunction of keratinocyte subpopulations may affect the occurrence and development of vitiligo.
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Narrowband ultraviolet B has been applied to the treatment of vitiligo for more than 10 years in China.Currently,there are no consistent standards for clinical treatment parameters,and patients cannot benefit from non-standard treatment,which is liable to cause erythema,blisters,photoaging and other adverse reactions.Based on the Vitiligo Working Group recommendations for narrowband ultraviolet B phototherapy for vitiligo,relevant literature and clinical experiences,the authors discuss parameters of narrowband ultraviolet B phototherapy for vitiligo from the aspects of treatment frequency,initial dosing,dose adjustment during consecutive treatment or after missed treatment,response plateau,treatment course and maximum acceptable number of phototherapy,so as to improve the efficacy of narrowband ultraviolet B phototherapy for vitiligo.
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Objective To evaluate the efficacy of long-term treatment with tacrolimus ointment in patients with vitiligo.Methods A total of 156 patients with vitiligo (70 males and 86 females) were enrolled into this study,who were firstly diagnosed with vitiligo and continuously followed up in Department of Dermatology,Yiwu Dermatology Hospital and the Third People's Hospital of Hangzhou between January 2016 and February 2018.Of the 156 patients,114 were adults,and 42 were children aged 6-18 years.All the patients received 6-month treatment with tacrolimus ointment twice a day.They were followed up once a month,the time to initial repigmentation and vitiligo area severity index (VASI) were recorded,and VASI improvement rate was calculated.After 6-month treatment,the patients achieving marked improvement were divided into conventional treatment group and interval treatment group to be treated with tacrolimus ointment twice a day and once every 3 days respectively for another 6 months,and final efficacy of the 2 protocols was compared.Mauchly's test of sphericity,randomized block analysis of variance and Bonferroni method were used to analyze differences among pre-treatment VASI scores,and 1-to 6-month post-treatment VASI scores.Results Among the 156 patients,the pre-treatment,and 1-,2-,3-,4-,5-and 6-month post-treatment VASI scores [M(P25,P75)] were 2 (0.6,5),2 (2,5),1.6 (0.575,4.5),1.2 (0.5,4.0),0.4 (1.15,3.5),0.75 (0.3,2.575),0.6 (0.2,2.2) respectively.Additionally,there was a significant difference in the VASI score among the above time points (F =6.14,P ＜ 0.05),and the VASI score showed a gradual decreasing trend over time.During the initial 6-month treatment,the average time to initial repigmentation was 2.2 ± 0.6 months.Of the 156 patients,98 (62.8％) received marked improvement,and the time to marked improvement was 5.2 ± 0.6 months.Of the 98 patients who received marked improvement,46 completed another 6-month treatment.After 12-month treatment,there was no significant difference in the repigmentation rate between the conventional treatment group (73.3％ ± 18.2％) and interval treatment group (72.0％ ± 16.2 ％,t =0.42,P ＞ 0.05).Conclusions The initial 6-month topical tacrolimus therapy showed marked efficacy for vitiligo.After the marked improvement of skin lesions,good efficacy can be maintained by long-term treatment with topical tacrolimus once every 3 days or twice a day.
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Objective:To investigate the clinical effect of Loquat Lung-Cleaning Decoction combined with fusidic acid cream in the treatment of acne vulgaris.Methods:From March 2019 to November 2019, 50 patients (9 males and 21 females, aged 13-31 years) with acne vulgaris were collected from the acne clinic of the dermatology department of the Hospital and divided into the experimental group and the control group with 25 cases in each group. The control group was treated with fusidic acid cream alone, and the experimental group was treated with Loquat Lung-Cleaning Decoction combined with fusidic acid cream. Six weeks after treatment, the clinical effect and adverse reactions were observed.Results:In the experimental group, 7 cases were cured, 10 cases were markedly effective, 6 cases were effective, but 2 cases were ineffective; the apparent efficiency was 68%. In the control group, 4 cases were cured, 6 cases were markedly effective, 9 cases were effective, and 6 cases were ineffective; the apparent efficiency was 40%. The effective rate of treatment in the experimental group was significantly higher than that in the control group, and the area of skin lesions was significantly smaller than that in the control group. The difference was statistically significant (rank sum test Z=1.98442, P=0.0472<0.05, rate analysis χ2 test χ2=3.945, P<0.05). Conclusions:The treatment of acne vulgaris with Loquat Lung-Cleaning Decoction and fusidic acid cream is effective.
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Objective:To study the characteristics of intestinal microflora in patients with vitiligo, and to analyze the relationship between the changes of intestinal microflora and the incidence of vitiligo, so as to provide new ideas for clinical treatment.Methods:Fecal specimens from 30 patients with vitiligo and 30 healthy adults were collected and analyzed qualitatively by Roche/45 high-throughput sequencing platform. At the same time, macrogenomics was used to analyze the feces of 5 patients with vitiligo and 5 healthy adults to identify the potential regulatory pathways.Results:The bacterial species in the feces of patients with vitiligo were similar to those of healthy people, but the intestinal microbial diversity of patients with vitiligo was significantly reduced ( P<0.01); the abundance of Proteus and Clostridium was significantly reduced at phylum level; at genus level, 7 of them were Bacteroides, Escherichia coli Shigella, Rochella, Bacillus anthracis, Clostridium clostridium, Jordani bacteria. The abundance of RF9 and Prunella-7 decreased significantly ( P<0.01), while the abundance of 4 genera (Rumen Coccus-1, Rumen Coccus UCG, Trichomonas and Streptococcus) increased significantly ( P<0.01). The expression of Streptococcus and Phase Anthraceae in vitiligo patients was significantly different: the former increased by 10.8 times, the latter decreased by 6.517 times, and an intestinal microorganism based on 11 vitiligo-related genera was constructed. The random forest model of bacterial flora showed that AUC of the discriminant model was 0.89 in ROC, and macrogenomic analysis showed that the disorders of vitiligo-related bacterial flora were mainly related to immune-related pathways (such as WNT pathway, Notch pathway), energy metabolism, mitochondrial function and amino acid metabolism (such as phenylalanine metabolism). Conclusions:The diversity of bacterial community in intestinal microecological environment of vitiligo patients is significantly different from those in normal people. The imbalance of bacterial community may be involved in the pathogenesis and development of vitiligo. Supplementation of probiotics may be beneficial to the treatment of vitiligo.
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Objective To investigate the reflectance confocal microscopic features of primary cutaneous amyloidosis.Methods A total of 30 outpatients with confirmed primary cutaneous amyloidosis were enrolled from Department of Dermatology,Hangzhou Third People's Hospital between June 2015 and October 2016.Their lesions were observed by reflectance confocal microscopy (RCM),and subjected to histopathological examination.Results RCM imaging of primary cutaneous amyloidosis lesions showed hyperkeratosis in 22 cases,acanthosis in 15,hyperpigmentation in the basal cell layer in 20,incontinence of pigment in the dermal papilla and superficial dermis in 20,and enlarged polymorphous dermal papilla with highly refractive clumps in 30.These RCM features were highly consistent with histopathological findings.Conclusion The typical RCM characteristic of primary cutaneous amyloidosis is highly refractive clumps in the dermal papilla,and RCM may serve as a non-invasive tool for assisting diagnosis of primary cutaneous amyloidosis.
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Objective To evaluate the association between clinical efficacy of and response to narrow-band ultraviolet B (NB-UVB) phototherapy in patients with confirmed vitiligo,and to identify factors influencing phototherapy response.Methods From January 2017 to March 2018,a total of 104 patients with vitiligo,who received NB-UVB phototherapy at the Department of Dermatology,Hangzhou Third Hospital,Zhejiang University of Traditional Chinese Medicine,were enrolled into this study.Among them,there were 55 males and 49 females,with an average age of 32.06 years (range:4-64 years).Their course of disease ranged from 2 months to 27 years,and the average course of disease was 8.15 years.The phototherapy sessions required to achieve initial repigmentation and to enter the plateau phase during which repigmentation ceased to increase with treatment sessions (plateau photothereapy sessions),as well as effective phototherapy sessions (the difference between the phototherapy sessions required to achieve initial repigmentation and those to enter the plateau phase) were recorded.Rank sum test was used to analyze the difference in the above phototherapy sessions between patients with different response,and multiple linear regression analysis to identify factors influencing plateau photothereapy sessions and effective phototherapy sessions.Results All the 104 patients were followed up.Among them,2 patients achieved complete repigmentation,1 showed no response to the phototherapy,and the remaining 101 patients were enrolled into the analysis.Among the 101 patients,52 achieved ≥ 50％ repigmentation (high efficacy group),and 49 achieved ＜ 50％ repigmentation (low efficacy group).Compared with the low efficacy group,the high efficacy group showed significantly less phototherapy sessions required to achieve initial repigmentation (M [P25,P75],8 [7,10] vs.10[8,13],Z =3.125,P =0.002),but significantly more plateau phototherapy sessions (41 [29,60] vs.35 [26,44],Z =-3.375,P ＜ 0.001) and effective phototherapy sessions (32 [18,51] vs.24 [8,36],Z =-4.407,P ＜ 0.001).Multiple linear regression analysis showed that the total number of plateau phototherapy sessions was positively correlated with vitiligo area (β =0.360,t =2.698,P =0.008) and duration of disease (β =0.215,t =2.325,P =0.022),but negatively correlated with vitiligo disease activity score (VIDA) (β =-2.665,t =3.969,P ＜ 0.001).Moreover,the number of effective phototherapy sessions was positively correlated with vitiligo area (β =0.358,t =2.582,P =0.011) and duration of disease (β =0.216,t =2.276,P =0.025),but negatively correlated with VIDA (β =-2.669,t =-3.935,P ＜ 0.001) and phototherapy sessions required to achieve initial repigmentation (β =-1.011,t =-4.508,P ＜ 0.001).Conclusions During the treatment with NB-UVB,the less the number of phototherapy sessions required to achieve initial repigmentation is,the more the number of effective phototherapy sessions is,and the better the efficacy of NB-UVB phototherapy is.Vitiligo area,duration of disease and clinical staging may provide the basis for predicting phototherapy sessions required to enter the plateau phase.