Variation of COL7A1 gene in dystrophic epidermolysis bullosa pruriginosa / 中华医学遗传学杂志

OBJECTIVE@#To perform gene mutation analysis in a Chinese pedigree with dystrophic epidermolysis bullosa pruriginosa (DEB-Pr), and explore phetotype, genotype, and genotypes-phenotypes relationship of DEB-Pr.@*METHODS@#Potential variants of the COL7A1 gene were detected by skin targeted sequencing panel and verified by Sanger sequencing. The pathogenicity of the variation was analyzed.@*RESULTS@#Compound heterozygous variants, c.4128delT and c.8234G>A, were detected in the COL7A1 gene of the two patients. The c.4128delT(p.Pro1376fs) variant was derived from their mother and unreported previously. According to the American College of Medical Genetics and Genomics Standards and Guidelines, it was suggested to be a pathogenic mutation. The c.8234G>A(p.Arg2745Gln) variant was derived from their father, and possibly is a pathogenic variation.@*CONCLUSION@#In this study, the compound heterozygous variants of c.4128delT(p.Pro1376fs) and c.8234G>A(p.Arg2745Gln) of the COL7A1 gene probably underlies the disease in this patient and his sister. And our study expands the database on mutations of DEB-Pr.

Mutation analysis of the ADAR gene in two cases of dyschromatosis symmetrica hereditaria and a survey of their families / 中华皮肤科杂志

Objective:To investigate two Chinese pedigrees with dyschromatosis symmetrica hereditaria (DSH) , and to analyze gene mutations in the pedigrees.Methods:Clinical data were collected from two probands with DSH and other family members in their pedigrees. Peripheral blood samples were obtained from the two probands, their parents and 100 unrelated healthy controls. Gene mutations were detected by using a skin-targeted sequencing panel, and then verified by Sanger sequencing.Results:Case 1, an 18-year-old male patient, presented with millet-sized hyperpigmented and hypopigmented macules scattered on the dorsum of both hands and feet at the age of 5 years, and his mother had similar manifestations. A novel heterozygous frameshift mutation c.1970dupT (p.F657fs) was identified in exon 5 of the ADAR gene in case 1 and his mother, but not found in his father. Case 2, an 8-year-old male patient, presented with mottled rice- to soybean-sized brown hyperpigmented macules and hypopigmented macules on the face and neck, lower back, buttocks, lower limbs, as well as hands and feet, and his father presented with similar manifestations. A known heterozygous frameshift mutation c.2433_2434delAG (p.T811fs) was identified in exon 7 of the ADAR gene in case 2 and his father, but not found in his mother. Neither of the two mutations was identified in the 100 unrelated healthy controls.Conclusion:In this study, a novel mutation c.1970dupT (p.F657fs) in the ADAR gene was identified in a patient with DSH.

Gene mutation analysis in a Chinese pedigree with autosomal recessive woolly hair / 中华皮肤科杂志

Objective:To identify causative genes for autosomal recessive woolly hair (ARWH) in a family.Methods:Clinical data were collected from two patients and other family members in a Chinese pedigree of Han nationality with ARWH. Peripheral blood samples were obtained from the two patients, their unaffected parents and 100 unrelated healthy individuals, and DNA was extracted from the blood samples. A next-generation skin-targeted sequencing panel was used to detect gene mutations in the patients, and Sanger sequencing was performed to verify the sequencing results. The function of protein encoded by the mutant gene was predicted.Results:Two missense mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) were both identified in the LIPH gene of the two patients, which were inherited from their father and mother respectively. Neither of the two mutations was identified in the 100 unrelated healthy controls. Interspecies sequence alignment showed that leucine at amino acid position 177 and cysteine at amino acid position 246 of the protein encoded by the LIPH gene were highly evolutionarily conserved. As SIFT and Polyphen-2 softwares showed, the mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) were both predicted to be detrimental variations.Conclusion:Two missense mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) in the LIPH gene may contribute to the clinical phenotype of the two patients with ARWH in this family.

Genetic testing and genotype-phenotype analysis for a child with X-linked hypohidrotic ectodermal dysplasia / 中华医学遗传学杂志

OBJECTIVE@#To carry out genetic testing for a Chinese patient with X-linked hypohidrotic ectodermal dysplasia (XLHED) and explore its genotype-phenotype correlation.@*METHODS@#Clinical data of the patient was collected. Peripheral blood samples were taken from the patient, his parents and 100 unrelated healthy controls. Genetic variants were detected by using next-generation sequencing using a skin-disease panel through targeted capture and next generation sequencing. Candidate variant was verified by Sanger sequencing. All literature related to genetic testing of XLHED patients in China was searched in the database, and the genotypes and phenotypes of patients in the literature and the correlation between them were statistically analyzed.@*RESULTS@#A novel splice site variant c.655_689del was detected in the patient but not among his parents and the 100 unrelated healthy controls. So far 61 variants of the EDA gene have been identified among Chinese patients with XLHED, which suggested certain degree of genotype-phenotype correlation.@*CONCLUSION@#A novel c.655_689del variant has been identified in the EDA gene, which has expanded the spectrum of EDA gene variant and facilitated delineation of the genotype-phenotype correlation of XLHED.

Two cases of Darier′s disease: ATP2A2 gene mutation analysis and a family survey / 中华皮肤科杂志

Case 1, a 16-year-old male patient, presented with local verrucous hyperplasia and clustered brown follicular keratotic papules on the face, neck and bilateral axillae, some of which merged into plaques; his mother had similar medical history and clinical manifestations. Case 2, a 21-year-old male patient, presented with local verrucous hyperplasia and diffuse follicular keratotic papules on the head, face, neck, trunk, bilateral axillae and buttocks, some of which merged into patchy lesions; none of his family members had similar symptoms. Histopathological examination of the neck skin lesion of the case 2 showed epidermal hyperkeratosis with focal parakeratosis, suprabasal cleft with acantholysis, "villi" , corps ronds and grains in the spinous layer, and inflammatory cells infiltrating the superficial dermis. Genetic testing showed a missense mutation c.2300A>G in exon 15 of the ATP2A2 gene in both the case 1 and his mother, and a splice-region mutation c.2097+5G>A at the junction between exon 15 and intron 15 of the ATP2A2 gene in the case 2. Neither of these mutations were identified in the other family members of the 2 patients.

Seven cases of vitiligo caused by eyebrow tattooing / 中华皮肤科杂志

Objective:To report 7 cases of vitiligo caused by eyebrow tattooing.Methods:Seven cases of vitiligo caused by eyebrow tattooing were collected from Department of Dermatology, Henan Provincial People′s Hospital from December 2017 to May 2019, and their clinical features were retrospectively analyzed.Results:One month to 1 year after eyebrow tattooing, several eyebrows became white in the 7 patients. In the early stage, only several eyebrows became white, and the surrounding skin was normal, but white patches with unclear boundaries gradually appeared around the eyebrows in the later stage. Reflectance confocal scanning microscopy of skin lesions on the eyebrow showed depigmentation in the basal layer and around hair follicles, and highly refractive amorphous substances (colorants) in the superficial and middle dermis. The 7 patients all showed negative patch test reactions to eyebrow colorants but positive reactions to sodium dodecyl sulfate.Conclusion:No depigmentation was observed on the eyebrow skin in the early stage of vitiligo caused by eyebrow tattooing, and reflectance confocal scanning microscopy of eyebrow lesions may be beneficial in reducing its misdiagnosis.

Mutation analysis in a pedigree with Rothmund-Thomson syndrome / 中华皮肤科杂志

Objective@#To detect gene mutations in a pedigree with Rothmund-Thomson syndrome (RTS) .@*Methods@#Clinical data were collected from two patients (an older sister and a younger brother) and their family members in a Chinese pedigree of Han nationality with RTS. Blood samples were obtained from the two patients, their unaffected older brother, their parents and 100 unrelated healthy controls. DNA was extracted, and all the exons in the encoding area of the RECQL4 gene were amplified by PCR. Gene mutations were detected by a skin-targeted next-generation sequencing panel, and verified by Sanger sequencing.@*Results@#Two heterozygous mutations were identified in the RECQL4 gene of the two patients, including a splice site mutation c.2886-1G>A and an insertion mutation c.1013_1014insC, which were inherited from the father and mother of the patients respectively. Meanwhile, neither of the two mutations was observed in 100 unrelated healthy controls or the older brother of the patients.@*Conclusion@#The splice site mutation c.2886-1G>A and the insertion mutation c.1013_1014insC in the RECQL4 gene may contribute to the clinical phenotype of the patients in this pedigree with RTS.

Mutation analysis in a pedigree with Rothmund-Thomson syndrome / 中华皮肤科杂志

Objective To detect gene mutations in a pedigree with Rothmund-Thomson syndrome (RTS).Methods Clinical data were collected from two patients (an older sister and a younger brother)and their family members in a Chinese pedigree of Han nationality with RTS.Blood samples were obtained from the two patients,their unaffected older brother,their parents and 100 unrelated healthy controls.DNA was extracted,and all the exons in the encoding area of the RECQL4 gene were amplified by PCR.Gene mutations were detected by a skin-targeted next-generation sequencing panel,and verified by Sanger sequencing.Results Two heterozygous mutations were identified in the RECQL4 gene of the two patients,including a splice site mutation c.2886-1G＞A and an insertion mutation c.1013_1014insC,which were inherited from the father and mother of the patients respectively.Meanwhile,neither of the two mutations was observed in 100 unrelated healthy controls or the older brother of the patients.Conclusion The splice site mutation c.2886-1G＞A and the insertion mutation c.1013_1014insC in the RECQL4 gene may contribute to the clinical phenotype of the patients in this pedigree with RTS.

Analysis of keratin 1 gene mutation and phenotypes in a pair of twins with bullous congenital ichthyosiform erythroderma / 中华皮肤科杂志

Objective To identify mutations in keratin genes (KRT1 and KRT10) in a pair of twins with bullous congenital ichthyosiform erythroderma (BCIE),and to explore the relationship between the causative genes and phenotypes.Methods Clinical data were collected from a pair of twins with BCIE and their family members.Peripheral blood samples were obtained from the twins,their old brother and parents,and DNA was extracted from these blood samples.Polymerase chain reaction (PCR)was performed to amplify all the coding exons and their flanking sequences of the KRT1 and KRT10 genes,and 100 unrelated healthy persons served as controls.Results The 11-year-old male proband presented with recurrent blisters,hypertrophy and desquamation all over the body for 11 years.His twin brother had similar skin lesions.Skin examination of the proband showed diffuse erythema covered with thick scaly crusts on the trunk and extremities.Blisters,bullae and erosions due to ruptured blisters were observed locally with tenderness on palpation.There were obvious hyperkeratotic and hard lesions on the big joints of the extremities.Diffuse hyperkeratosis could be seen on the palms and soles.A mutation c.591 + 1G ＞ A was identified at position 1 in intron 1 of the KRT1 gene in the twins,but not in the 3 healthy family members or the 100 unrelated healthy controls.Conclusion The mutation c.591 + 1G ＞ A at position 1 in intron 1 of the KRT1 gene may contribute to the clinical phenotype of the twins with BCIE.

Cutaneous mucormycosis caused by Mucor irregularis: a case report / 中华皮肤科杂志

A 39-year-old female patient presented with a painful plaque with ulcers on the right cheek for 2 months.She had acute myeloid leukemia for 1 year.After treatment,the patient achieved remission,but experienced recurrence half a year prior to the presentation.Skin examination showed a violaceous plaque measuring 5 cm × 5 cm in size on the right cheek with erosions and ulcers in the center,whose surface was covered with yellowish brown crusts.Granulation tissues were observed on the plaque,and yellow pus was exuded after the crusts were removed.The boundary of the plaque was sharp and slightly elevated,and there was obvious tenderness on palpation.Laboratory examination revealed increased white blood cell (WBC,28.75 × 109/L) and lymphocyte counts (27.17 × 109/L),but decreased neutrophil (1.05 × 109/L) and red blood cell counts (2.20 × 1012/L),hemoglobin level (69 g/L) and platelet count (84 × 109/L) in the peripheral blood.The hepatic and renal function,electrolyte level and electrocardiogram were normal.Hematoxylin and eosin (HE) staining and periodic acid-Schiff staining of the lesion showed a large number of lymphocytes and histiocytes infiltrating in the dermis and broad aseptate hyphae.The fungal microculture yielded broad hyalinea septate hyphae,fungal rhizoids,stolons and spherical sporangia.The isolated fungus was identified as Mucor irregularis by using molecular biology techniques.The patient was diagnosed with primary cutaneous mucormycosis caused by Mucor irregularis complicated by acute myeloid leukemia.Then,the patient was treated with oral hydroxyurea at a dose of 0.5 g thrice a day,a single-dose intravenous infusion of 4 units of red blood cell suspension,and intravenous drip infusion of amphotericin B at an initial dose of 5 mg/d,which increased by 5 mg every day until 25 mg/d (about 0.5 mg· kg-1· d-1).After the treatment,the lesion gradually became fiat and smaller.After 12-day treatment,the patient was discharged because of a certain reason,and finally lost to follow-up.

Gravitational erythema：a case report / 中华皮肤科杂志

A 15-year-old female patient presented with orthostatic erythema in the limbs for half a year. Skin examination revealed that diffuse reticular erythema appeared in the limbs after standing with her arms hanging down for 30 seconds, while the erythema disappeared and skin color returned to normal after raising her arms or lying flat for 30 seconds. The rash in the limbs was reproduced by inflating a sphygmomanometer cuff to 60 mmHg for 2 minutes with the limbs in the horizontal position. Antinuclear antibodies were positive at a titer of 1 ∶1 000. Histopathological examination showed dilated small vessels in the superficial dermis without obvious inflammatory cell infiltration. The patient was diagnosed with gravitational erythema.

Clinical efficacy of 5-aminolevnlinic acid-based photodynamic therapy for the treatment of lichen sclerosus et atrophicus / 中华皮肤科杂志

Objective To estimate the efficacy and safety of 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) in the treatment of lichen sclerosus et atrophicus of the vulva.Methods An open and noncontrolled clinical study was performed.Forty-two patients were enrolled in this study and received ALA-PDT once every two weeks for 2 to 4 times.Follow-up visits were arranged at 2,4 and 8 weeks after initiation of treatment,and patients were evaluated at the baseline (before treatment) and all the follow-up time points for the efficacy and safety of treatment.Results Finally,38 patients completed the trial and 4 patients were lost to follow up.The total response rate was 81.6％ (31/38) at the end of the treatment.The average symptom and sign score in these patients was significantly lower at 2,4 and 8 weeks after initiation of treatment than that before treatment (17.6 ± 10.18,11.6 ± 8.35 and 7.6 ± 5.93 vs.29.3 ± 9.17,t =5.26,8.80,12.22,respectively,all P ＜ 0.01).A significant improvement was also observed in the other aspects,such as skin lesion area,hypopigmentation,erosion/rhagades and itching score at 2,4 and 8 weeks,as well as in skin atrophy at 8 weeks after initiation of treatment compared with those before treatment (all P ＜ 0.05).Local burning sensation was the main adverse reaction to ALA-PDT,and 16％ (6/38) of these patients complained of severe pain during the first treatment.Conclusion ALA-PDT shows favorable efficacy in patients with lichen sclerosus et atrophicus of the vulva with a rapid onset of action.

Quantification of CD123+ BDCA-2+ plasmacytoid dendritic cells in skin lesions and peripheral blood of patients with psoriasis vulgaris / 中华皮肤科杂志

[Objective] To quantify CD123+ BDCA-2+ plasmacywid dendritic cells (PDCs) in skin lesions and peripheral blood of patients with psoriasis vulgaris,and to investigate their significance.[Methods] Skin tissue samples were resected from the lesions of 22 patients with psoriasis vulgaris and normal skin of 15healthy controls,and peripheral blood samples were also obtained from these subjects.Immunohistochemical technique (SP) was used to quantify PDCs in skin tissue samples,and flow cytometry to determine the proportion of PDCs in peripheral blood samples.[Results] Immunohistochemical study showed that the density of PDCs in the psoriatic lesions was significantly higher than that in the healthy controls (( 10.1 ± 2.1 )/mm2 vs.(0.4 + 0.6)/mm2,t =17.34,P ＜ 0.01 ).On the contrast,the proportion of PDCs in peripheral blood was 0.17％ ±0.07％ in the patients with psoriasis vulgaris,significantly lower than that in the healthy controls (0.33％ ±0.20％,t =4.48,P ＜ 0.01 ).[Conclusion]s The proportion of PDCs is reduced in the peripheral blood of patients with psoriasis vulgaris,which may be related to the initiation and progression of psoriasis vulgaris.

Effects of a short hairpin RNA targeting STAT3 gene on the biological behavior of a human malignant melanoma cell line A375 / 中华皮肤科杂志

Objective To study the effects of short hairpin RNA (shRNA) targeting STAT3 gene on the proliferation and apoptosis of A375 human malignant melanoma cells.Methods Sense and antisense oligonucleotides with small hairpin structures targeting STAT3 gene were designed,synthesized and cloned into the plasmid vector psiRNA-hHlneo after annealing.Cultured A375 cells were divided into 3 groups: control group receiving no treatment,psiRNA-H1 group transfected with empty plasmid,and psiRNA-H1/STAT3 group transfected with the recombinant plasmid containing the shRNA.After additional culture for different durations,reverse transcription PCR and Western blot were performed to detect the expression of STAT3 mRNA and protein,methyl thiazolyl tetrazolium (MTT) assay to evaluate cell proliferation,flow cytometry to assess cell cycle and apoptosis.Results The expression level of STAT3 mRNA and protein in A375 cells in psiRNA-H1/STAT3 group (0.2136 ± 0.0626,0.8214 ± 0.043,respectively) were significantly lower than that in the control group (0.7826 ± 0.0701,3.1693 ± 0.0846,respectively,both P ＜ 0.01) and psiRNA-H1 group (0.8518 ± 0.0783,3.218 ± 0.078,respectively,both P ＜ 0.01 ).The inhibition rates of cell proliferation at 24,48 and 72 hours were 21.35％ ± 2.12％,32.52％ ± 2.64％ and 40.4％ ± 3.08％ respectively in psiRNA-H1/STAT3 group,statistically different from those in the control group (1.39％ ± 0.53％,3.05％ ± 1.16％,4.41％± 1.42％,respectively,all P ＜ 0.01) and psiRNA-H1 group (2.63％ ± 1.38％,5.84％ ± 2.47％,10.32％ ±2.48％,respectively,all P ＜ 0.01).Flow cytometry showed a statistical increase in cell apoptosis rate in psiRNA-H1/STAT3 group compared with the control and psiRNA-H1 group (81.06％ ± 2.10％ vs.26.28％ ±0.47％ and 27.31％ ± 1.05％,both P ＜ 0.01 ).The psiRNA-H1/STAT3 group exhibited a higher percentage of cells at G0/G1 phase (68.43％ ± 4.00％) but a lower percentage of cells at S phase (17.4％ ± 2.05％) compared with the control group (60.07％ ± 2.47％,P ＜ 0.05; 28.40％ ± 2.09％,P ＜ 0.01 ) and psiRNA-H1 group (60.29％ ± 2.26％,27.34％ ± 3.63％,both P ＜ 0.05 ).Conclusions The small interference RNA targeting STAT3 gene can specifically down-regulate the expressions of STAT3 mRNA and proteins in,inhibit cellular proliferation of,and induce apoptosis in,A375 cells.

Flashlamp-pumped pulsed dye laser (595 nm) for the treatment of port-wine stains: a clinical review of 1560 patients / 中华皮肤科杂志

Objective To evaluate the efficacy and safety of a 595-nm flashlamp-pumped pulsed dye laser in the treatment of patients at different ages with port-wine stains (PWS).Methods A retrospective review was performed in 1560 patients with PWS who had been treated with a 595-nm flashlamp-pumped pulsed dye laser.Treatment parameters were selected according to the age of and types of lesions in patients.Results The total response rate was 76.73％ (1197/1560) in all of the patients.Clinical efficacy of the flashlamppumped pulsed dye laser was closely correlated with patients'age (x2 =83.47,P ＜ 0.01) and types of lesions (x2 =46.30,P ＜ 0.01 ).There was a low incidence of adverse reactions which were well tolerable.Conclusion The 595-nm flashlamp-pumped pulsed dye laser is safe and effective for the treatment of PWS.

Multiple keratoacanthoma centrifugum marginatum: first case report in China / 中华皮肤科杂志

A case of multiple keratoacanthoma centrifugum marginatum is first reported in China. A 37-year-old woman was admitted to the hospital for papules and plaques on her face, which had been increasing in number for 4 months. Cutaneous examination revealed dozenes of well-marginated, pale-red or skin-colored crateriform papules of variant size, and plaques in a geographic pattern on her face. The papules presented with a central umbilication filled with grey-brown corneous material. The plaques were surrounded by dyke-like borders, covered with thick, crusted brown corneous material, and partly depressed in the center. Histopathology showed hyperkeratosis, parakeratosis, irregular strip-like extension of epidermis into dermis, keratinous cysts and squamous eddies. The tumor cells had eosinophilic and glassy cytoplasm characteristic of keratoacanthoma.Given both the clinical and histologic evidence, a diagnosis of multiple keratoacanthoma centrifugum marginatum was made. After more than 3 months of treatment with oral acitretin and topical tretinoin, the lesions faded,leaving rugosity scars. No relapse was noted during 3-year follow-up.

Effects of genistein on biological activities of and collagen synthesis by human skin fibroblasts in vitro / 中华皮肤科杂志

Objective To study the effects of genistein on the biological characteristics of and collagen synthesis by human skin fibroblasts in vitro.Methods Human skin fibroblasts(HSFBs)were obtained from the prepuce of healthy adolescents,and suhjected to primary culture.Atier 5 to 15 passages of culture,HSFBs were treated with various concentmtions(0.03125,0.0625,0.125,0.25,0.5,1mg/L)of genistein for different durations.The potential of cell proliferation was detected by MTT assay and growth curves were drawn for HSFBs.Flow cytometry(FCM)and RT-PCR were used to estimate cell cycle phases and mRNA expression of type Ⅰ procollagen.respectively.Results The proliferation rate of HSFBs was 97.7%,113.8%,132.5%,116.4%,94.5%and 83.3%after treatment with genistein of 0.03125,0.0625,0.125,0.25,0.5 and 1 mg/L,respectively.The genistein of more than 0.5 mg/L displayed an inhibitive effect on the proliferation of HSFBs.while that between 0.0625 and 0.25 mg/L showed a promotive effect.Atier treatment with genistein at 0.0625,0.1 25 and 0.25 mg/L,the percentage of HSFBs in S phase and G2 phase significantly increased compared with untreated HSFBs(S phase:41.15%±2.88%,61.89%±3.16%,48.18%±1.68%vs30.12%±0.60%,P＜0.05;G2 phase:9.76%±3.99%,10.40%±0.54%,7.46%±2.47%vs 0.61%±0.16%,P＜0.05).Compared with the untreated HSFBs.the relative mRNA expression level of type Ⅰ procollagen was increased with genistein of 0.0625,0.125 and 0.25 mg/L(0.4814±0.0138,0.5767±0.0291,0.5675±0.0272 vs 0.4101±0.0236,P＜0.01),but decreased with genistein of Ⅰ and 0.5 mg/L(0.1662±0.0165 and 0.2017±0.0203 vs 0.4101±0.0236,P＜0.01).ConclusionCertain concentrations of genistein could enhance the proliferation and growth of as well as mRNA expression of type Ⅰ procollagen in HSFBs.