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COVID-19 can be accompanied by a variety of cutaneous abnormalities, which mainly include vascular lesions (chilblain-like lesions, livedo reticularis, purpura, ecchymosis, acral cyanosis, gangrene, etc) and inflammatory lesions (diffuse erythema, morbilliform exanthem, acute urticaria, varicella-like exanthem, etc) . Some types of skin lesions may be the first symptom or the only clinical manifestation of COVID-19.
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Objective:To investigate the effect of KRT5 knockdown in keratinocytes on melanin content in co-cultured melanocytes, and to explain mechanisms underlying formation of hyperpigmented lesions in reticulate pigmented anomaly of the flexures (Dowling-Degos disease, DDD) .Methods:HaCaT cells with heterozygous mutations in the KRT5 gene were obtained by using clustered regularly interspaced short palindromic repeats (CRISPR) -CRISPR-associated protein 9 (Cas9) technology (experimental group) , and HaCaT cells transfected with non-targeting single guide RNA:Cas9 protein complex served as control group, both of which were in vitro co-cultured with primary human melanocyte cells (HEMn) separately. Immunofluorescence study was conducted to determine the expression of cytokeratin and melanosomes in co-cultured cells; melanin content was detected in melanocytes in different co-culture groups, which were obtained by differential trypsinization. Immunohistochemical study was performed to determine the expression of melanocyte-specific premelanosome protein 17 (Pmel17) in skin lesions in a patient with DDD carrying a KRT5 mutation and normal skin tissues in a healthy control. Results:Sanger sequencing showed a heterozygous mutation (c.1delA) at the initiation codon of exon 1 of the KRT5 gene in HaCaT cells in the experimental group, but no mutation in the KRT5 gene in the control group. Western blot analysis showed that the KRT5 protein expression was significantly lower in the experimental group (0.60 ± 0.05) than in the control group (1.00 ± 0.00, t = 32.38, P = 0.001) . Compared with the co-culture system in the control group, the number of Pmel17-labeled melanosomes markedly increased with the melanin content elevated by 52.5% ( t = -3.48, P = 0.025) in the HEMn cells co-cultured with HaCaT cells in the experimental group. Immunohistochemical study showed that the Pmel17 expression increased in the skin lesions in the DDD patient with KRT5 mutation compared with the normal skin tissues in the healthy control. Conclusion:The effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on the co-cultured HEMn melanocytes was verified by the successfully established in vitro co-culture system, which provides a primary cell model for further studies on interaction mechanisms between keratinocytes and melanocytes, and on pathogenesis of skin pigmentation abnormalities.
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Objective:To investigate clinicopathological features and prognosis of transformed mycosis fungoides (TMF) .Methods:A retrospective analysis was performed on clinicopathological data collected from 24 patients with TMF, as well as on flow cytometry results of 16 peripheral blood samples obtained from 11 of the 24 patients, who visited Hospital of Dermatology, Chinese Academy of Medical Sciences between 2014 and 2020.Results:Among the 24 patients, 11 were males and 13 were females. Their average age at diagnosis of TMF was 50.0 years (range: 18 - 77 years), and patients with early-stage TMF (9 cases) and tumor-stage TMF (15 cases) were aged 44.8 and 52.6 years on average, respectively. The average time interval from diagnosis of MF to large cell transformation was 3.7 years, and 8 patients were diagnosed with TMF at the initial visit. Histopathologically, large cells infiltrated in a diffuse pattern in 20 cases, as well as in a multifocal pattern in 4, and the proportion of large cells in 7 cases was greater than 75%. Immunohistochemically, 18 patients showed positive staining for CD30, and the proportion of CD30-positive large cells was greater than 75% in 9; negative staining for CD30 was observed in 6. Flow cytometry of 16 peripheral blood samples showed the presence of cell subsets expressing clonal T cell receptor (TCR) -vβ in 2 of 4 patients with early-stage TMF and 10 of 12 with tumor-stage TMF, and tumor cells with higher forward scatter than normal lymphocytes were detected in 16 samples. During the follow-up, among the patients with early-stage TMF, 3 progressed to tumor-stage TMF 3.3 years on average after large cell transformation, 1 progressed to erythrodermic MF in stage IIIA, and the other 4 still showed an indolent course; among the patients with tumor-stage TMF, 1 progressed to stage-IV TMF, and 5 died 3.3 (1.5 - 6) years after large cell transformation.Conclusion:Large cell transformation may occur in patients with MF in any stage, some patients have poor prognosis, so close follow-up is needed for patients with TMF.
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Objective To analyze clinical and pathological features of papular elastorrhexis.Methods Clinical data were collected from 22 patients with confirmed papular elastorrhexis in Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Pekin Union Medical College from September 2006 to May 2018.Clinical manifestations,pathological findings and follow-up results were retrospectively analyzed.Results The average age of onset of the 22 patients was 5.7 years (range:1-10 years),and the male to female ratio was 4.5:1.The average duration from the occurrence of disease to the confirmation of diagnosis was 1.5 years,and no definite etiology was found.The patients had no itching or pain sensation.Skin lesions were soft,slightly elevated,well-circumscribed,round,oval or polygonal-shaped,white papules with diameters of 1-10 mm,and wrinkles appeared on the surface of the papule when the papule was pushed towards its center.Among the 22 patients,16 (73％) presented with scattered lesions,13 (59％)had less than 5 papules,and lesions were located in the trunk in 21 (95％).Histopathological examination of skin lesions in 8 patients showed no obvious increase of collagen fibers in the superficial and middle dermis,which were normally arranged with slightly widened spaces between them.Elastic fiber staining showed that elastic fibers disappeared or were dissociated focally in the superficial and middle dermis.After confirmed diagnosis,the 22 patients received no treatment.In 18 patients,skin lesions did not continue to expand after onset,and no new skin lesions occurred.Skin lesions were slightly enlarged,but remained steady thereafter in 4 patients.Sixteen patients achieved partial remission.Conclusions Papular elastorrhexis is a rare skin disorder of elastic fibers that occurs predominantly during childhood and adolescence,and its diagnosis relies on clinical manifestations combined with histopathological findings.No special treatment is needed and the prognosis is good.
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Objective@#To analyze clinical and pathological features of papular elastorrhexis.@*Methods@#Clinical data were collected from 22 patients with confirmed papular elastorrhexis in Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Pekin Union Medical College from September 2006 to May 2018. Clinical manifestations, pathological findings and follow-up results were retrospectively analyzed.@*Results@#The average age of onset of the 22 patients was 5.7 years (range: 1 - 10 years) , and the male to female ratio was 4.5∶1. The average duration from the occurrence of disease to the confirmation of diagnosis was 1.5 years, and no definite etiology was found. The patients had no itching or pain sensation. Skin lesions were soft, slightly elevated, well-circumscribed, round, oval or polygonal-shaped, white papules with diameters of 1 - 10 mm, and wrinkles appeared on the surface of the papule when the papule was pushed towards its center. Among the 22 patients, 16 (73%) presented with scattered lesions, 13 (59%) had less than 5 papules, and lesions were located in the trunk in 21 (95%) . Histopathological examination of skin lesions in 8 patients showed no obvious increase of collagen fibers in the superficial and middle dermis, which were normally arranged with slightly widened spaces between them. Elastic fiber staining showed that elastic fibers disappeared or were dissociated focally in the superficial and middle dermis. After confirmed diagnosis, the 22 patients received no treatment. In 18 patients, skin lesions did not continue to expand after onset, and no new skin lesions occurred. Skin lesions were slightly enlarged, but remained steady thereafter in 4 patients. Sixteen patients achieved partial remission.@*Conclusions@#Papular elastorrhexis is a rare skin disorder of elastic fibers that occurs predominantly during childhood and adolescence, and its diagnosis relies on clinical manifestations combined with histopathological findings. No special treatment is needed and the prognosis is good.
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With the development of medical technology,a new era of "big data" and "precision medicine" is coming.Artificial intelligence can assist in the diagnosis and treatment process,and reduce the pressure of data analysis on physicians.At present,artificial intelligence is mainly applied to image recognition,genetics and genomics,intelligent diagnosis and treatment,and prognosis prediction in medicine,whose accuracy may approach human experts.Image recognition is most studied,including skin image recognition.In non-image recognition field,artificial intelligence is less studied in dermatology.In the future,artificial intelligence will increase the efficiency of diagnosis and treatment,so as to benefit both physicians and patients.
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Objective To discuss the diagnosis and differential diagnosis of microcystic adnexal carcinoma (MAC).Methods Totally,10 patients with MAC were enrolled from Hospital for Skin Diseases,Chinese Academy of Medical Sciences from 2003 to 2017.Their clinical manifestations,histopathological and immunohistochemical features,treatment and prognosis were retrospectively analyzed.Results Of the 10 patients,3 were males and 7 were females.Their average age at the onset of MAC was 51.65 years.Skin lesions all occurred on the face,and on the upper lip in 6 cases.The lesions usually presented as solitary plaque or nodule,and ulceration occurred in 4 cases.Histopathologically,skin lesions consisted of epithelial cords with different numbers of keratinous cysts and tubular structures,and neural involvement occurred in 6 cases.However,mitotic figures were rare.Immunohistochemical staining showed epithelial cells and keratinous cysts stained positive for cytokeratin,as well as tubular structures and glandular cavities stained positive for carcinoembryonic antigen and epithelial membrane antigen.All the patients received surgical excision,and one patient experienced in situ recurrence 13 years later.No distant metastasis occurred in these patients.Conclusions MAC mainly presents as red plaques with occasional ulceration on the upper lip.Its definite diagnosis depends on characteristic histopathological changes in bidirectional differentiation into hair follicles and sweat glands,and immunohistochemical features are helpful to distinguish MAC from other adnexal tumors.
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Objective To analyze differentially expressed proteins in A375 melanoma cells before and after short hairpin RNA (shRNA)-mediated Cbl-b gene silencing.Methods The label-free quantitative proteomics approach was performed to identify differentially expressed proteins between A375 cells transfected with lentiviral vectors containing Cbl-b shRNA (Cbl-b shRNA group) and those with control lentiviral vectors (control group).Then,the properties of differentially expressed proteins were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) enrichment analysis.Western blot analysis was conducted to determine the expression of differential proteins (EphA2 and GSK3β) and phosphorylated protein kinase (p-AKT) after shRNA-mediated Cbl-b gene silencing.Statistical analysis was carried out by t test of two independent-samples for comparison of protein expression abundance between the two groups with SPSS 23.0 software.Additionally,the results of GO and KEGG enrichment analysis were analyzed by Fisher's exact test.Results A total of 3 449 proteins were identified and quantified,and 74 of them were differentially expressed between the Cbl-b shRNA group and control group.Compared with the control group,52 proteins were up-regulated and 22 were down-regulated in the Cbl-b shRNA group.GO enrichment analysis of differential proteins revealed that the top five significantly enriched biological processes were integrin-mediated cell adhesion,single-organism metabolic process,regulation of integrin-mediated cell adhesion,regulation of protein-targeting mitochondria and nucleic acid metabolic process.The top five significantly enriched molecular functions included DNA binding,2-iron,2-sulfur cluster binding,signaling receptor activity,cadherin binding and cell adhesion molecule binding.The top five significantly enriched cell components included nucleosome,DNA packaging complex,photoreceptor connecting cilium,DNA-protein complex and extracellular region part.KEGG enrichment analysis demonstrated that the top five significantly enriched melanoma-related signaling pathways were folate biosynthesis,axon guidance,extracellular matrix-receptor interaction,adherens junction and Wnt signaling pathways.As Western blot analysis revealed,the Cbl-b shRNA group showed lower protein expression of EphA2 (0.369),but higher protein expression of GSK3β (3.524) compared with the control group (1),which were consistent with the results of proteomics analysis.Additionally,the protein expression of p-AKT was down-regulated in Cbl-b shRNA group (0.453) compared with the control group (1).Conclusion Cbl-b may be involved in the occurrence of melanoma through a variety of biological pathways,and the EphA2/PI3K/AKT signaling pathway may be one important pathway.
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Objective To investigate clinical and pathological features of 6 cases of disabling pansclerotic morphea (DPM).Methods Clinical and pathological manifestations of and follow-up results in 6 patients,who were clinically and histopathologically diagnosed with DPM in the Department of Pathology,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College from 2007 to 2017,were retrospectively analyzed.Results Among the 6 patients,4 were male and 2 were female.The age of onset ranged from 3 to 10 years,with an average age of 6.5 years.The average duration from the occurrence to the confirmation of the diagnosis was 6.2 years (range,2-10 years).At all the lesional sites,skin atrophy,thining and tightness occurred,and the limbs became thin.Additionally,there were muscular atrophy and visible deep thick veins on the surface of the limbs.The contracture,deformity and dysfunction of the adjacent joints occurred in 4 cases,and the lower limbs were obviously shortened in 2 cases.Peripheral blood examination showed no increase of eosinophils or hypergammaglobulinemia.Imaging examination revealed smooth cortical bone and clear trabecular bone,and no osseous abnormality was observed.Histopathological examination of contracted skin lesions of the lower limbs revealed atrophic and thinned epidermis,hyperpigmentation in the basal layer,hyperplastic,thickened,hardened and partly homogenized collagen fibers in the middle to deep dermis,subcutaneous adipose tissue region and deep tissue of the skin.Conclusions DPM usually does not affect viscera,but often involves deep tissue of the limbs.Histopathologically,DPM is mainly characterized by obviously hyperplastic and hardened collagen fibers in the dermis and subcutaneous tissue.
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Objective To construct a recombinant lentiviral vector carrying the casitas B-lineage lymphoma (Cbl)-b short hairpin RNA (shRNA),and to evaluate its effect on the biological behavior of A375 melanoma cells in vitro.Methods Three specific shRNAs targeting Cbl-b gene and a negative control shRNA were designed and synthesized,and recombinant lentiviral vectors were constructed.A375 cells were divided into 5 groups to be transfected with 3 kinds of lentiviral vector expressing Cbl-b genespecific shRNAs (CBLB-shRNA-1 group,CBLB-shRNA-2 group and CBLB-shRNA-3 group),a lentiviral vector containing negative control shRNA (negative control group),and an empty vector (blank control group).Real-time fluorescence-based quantitative PCR and Western blot analysis were performed to determine the silencing efficiency at 72 hours after transfection.Cell counting kit-8 (CCK-8)assay was conducted to evaluate cellular proliferative activity at 24,48,72 and 96 hours after transfection,flow cytometry to detect cell apoptosis and cell cycle at 72 hours after transfection,and Transwell invasion assay to assess cellular invasive activity at 72 hours after transfection.Results Three recombinant lentiviral vectors containing Cbl-b shRNA were constructed successfully.As Western blot analysis revealed,the CBLB-shRNA-3 showed the highest silencing efficiency.CCK-8 assay indicated that the proliferative activity of A375 cells was significantly lower in the CBLB-shRNA-3 group than in the negative control group and blank control group at 72 and 96 hours after transfection(all P ＜ 0.01).Flow cytometry showed that the apoptosis rate of A375 cells was significantly higher in the CBLB-shRNA-3 group (22.73％ ± 6.58％) than in the negative control group (6.08％ ± 1.35％,P ＜ 0.01) and blank control group (6.34％ ± 1.07％,P ＜ 0.01).The CBLB-shRNA-3 group showed a significantly higher proportion of A375 cells at G1 phase,but a significantly lower proportion of A375 cells at S phase compared with the negative control group and blank control group(all P ＜ 0.01).Transwell assay showed that there were significant differences in the number of A375 cells crossing the artificial basement membrane (matrigel) at 72 hours after transfection among the negative control group,blank control group and CBLB-shRNA-3 group (76.60 ± 1.82,73.20 ± 3.83,19.60 ± 1.14,respectively;F =794.50,P ＜ 0.01).Conclusions A recombinant CBLB-shRNA-3-expressing lentiviral vector which can efficiently silence Cbl-b gene has been successfully constructed.It can inhibit the proliferation,cell cycle progression and invasive activity of A375 cells,but promote the apoptosis of A375 cells.
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A 59-year-old male patient presented with recurrent erythema and wheals all over the body for 6 years,complicated by irregular fever (the highest body temperature ＜ 39 ℃)and arthralgia for 2 months.He experienced a weight loss of 5 kg during two years prior to the presentation.Physical examination showed anemic comlexion,and there was no palpable enlargement of superficial lymph nodes,liver and spleen.Generalized erythema and wheals occurred all over the body,involving about 50％ of the body surface area.Histopathological examination of skin lesions showed infiltration of multiple neutrophilic granulocytes around blood vessels in the superficial dermis,and no vasculopathy was observed.Laboratory examinations revealed increased white blood cell (WBC) counts(13.97 × 109/L),erythrocyte sedimentation rate (ESR,136 mm/1 h) and C-reactive protein (CRP) level (75 mg/L),decreased hemoglobin level (96 g/L),and high serum levels of IgE (1 596 kU/L),IgG(20.4 g/L)and IgM(6 310 mg/L).Serum immunoelectrophoresis demonstrated that the IgM was a κ-type monoclonal immunoglobulin.Bone marrow examination showed active bone marrow hyperplasia.DNA sequencing analysis of 10 pairs of exons of the NLRP3 gene revealed 3 synonymous mutations,including c.663:C ＞ T:p.T221,c.732:G ＞ A:p.A244A and c.786:A ＞ G:p.R262R.Finally,the patient was diagnosed with Schnitzler's syndrome.There was no improvement of symptoms after the treatment with multiple oral antihistamines at increased dose levels.Then,the treatment protocol was adjusted to oral ciclosporin at a dosage of 200 mg/d for consecutive 18 days,but the patient still showed no response.After the treatment with oral prednisone (30 mg/d) and Tripterygium wilfordii tablets (66 μg thrice a day) for 2 weeks,the skin rashes subsided gradually,and arthralgia was relieved.Moreover,the WBC count,ESR and CRP level were decreased to 9.01 × 109/L,50 mm/1 h and 6 mg/L respectively,while the hemoglobin level was increased to 109.7 g/L.After 1-year follow-up,the dosage of glucocorticoids was gradually decreased to 8 mg/d.In addition,his condition was controlled well with no skin lesions and fever,except for occasional arthralgia in the knees and hip.
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Objective To explore whether paraquat (PQ) can induce the formation of neutrophil extracellular traps (NETs) in human peripheral blood.Methods Neutrophils were isolated from healthy human peripheral blood,and the cells were identified by hematoxylin-eosin (HE) strain.The cells were treated with different concentrations of PQ [0 (as control),200,400,600,800,1 000 and 1 200 μmol/L],and the cell viability was measured by cell proliferation and CCK-8 cytotoxicity detection kit,and the median lethal concentration of PQ was selected.The cells were treated with the median lethal concentration of PQ (PQ poisoning group),and the untreated cells were served as the control.Immunofluorescence staining was adopted to evaluate NETs formation.PicoGreen dye was used to determine the quantitative content of circulating free DNA.Western Blot was used to determine the expressions of citrullinated histone 3(H3Cit) and myeloperoxidase (MPO) in the supernatant.Results The purity of neutrophils was about 95％ by HE staining.The cells were treated with different concentrations of PQ,and the result showed that the viability of cells was (58 ± 2)％ with 800 μmol/L PQ for treatment.The immunofluorescence showed that there were few expressions of H3Cit and MPO in neutrophils in the control group,and there was no NETs formation,which was composed of DNA,H3Cit and MPO.Compared with the control group,a large amount of NETs was generated from neutrophils stimulated by 800 μmol/L of PQ.Meanwhile,quantitative result showed that the content of cell free DNA in the supernatant was significantly increased in PQ poisoning group as compared with that of control group (μg/L:2 235 ± 462 vs.561 ± 87,P ＜ 0.01).The protein expressions of H3Cit and MPO in the supernatant were also significantly increased as compared with those of control group [H3Cit protein expression (gray value):0.23 ± 0.03 vs.0.11 ± 0.01,MPO protein expression (gray value):0.47 ± 0.05 vs.0.21 ± 0.04,both P ＜ 0.05].Conclusion 800 μmol/L of PQ can induce the formation of NETs in human peripheral blood.
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A 59-year-old male patient presented with recurrent erythema and wheals all over the body for 6 years,complicated by irregular fever (the highest body temperature ＜ 39 ℃)and arthralgia for 2 months.He experienced a weight loss of 5 kg during two years prior to the presentation.Physical examination showed anemic comlexion,and there was no palpable enlargement of superficial lymph nodes,liver and spleen.Generalized erythema and wheals occurred all over the body,involving about 50％ of the body surface area.Histopathological examination of skin lesions showed infiltration of multiple neutrophilic granulocytes around blood vessels in the superficial dermis,and no vasculopathy was observed.Laboratory examinations revealed increased white blood cell (WBC) counts(13.97 × 109/L),erythrocyte sedimentation rate (ESR,136 mm/1 h) and C-reactive protein (CRP) level (75 mg/L),decreased hemoglobin level (96 g/L),and high serum levels of IgE (1 596 kU/L),IgG(20.4 g/L)and IgM(6 310 mg/L).Serum immunoelectrophoresis demonstrated that the IgM was a κ-type monoclonal immunoglobulin.Bone marrow examination showed active bone marrow hyperplasia.DNA sequencing analysis of 10 pairs of exons of the NLRP3 gene revealed 3 synonymous mutations,including c.663:C ＞ T:p.T221,c.732:G ＞ A:p.A244A and c.786:A ＞ G:p.R262R.Finally,the patient was diagnosed with Schnitzler's syndrome.There was no improvement of symptoms after the treatment with multiple oral antihistamines at increased dose levels.Then,the treatment protocol was adjusted to oral ciclosporin at a dosage of 200 mg/d for consecutive 18 days,but the patient still showed no response.After the treatment with oral prednisone (30 mg/d) and Tripterygium wilfordii tablets (66 μg thrice a day) for 2 weeks,the skin rashes subsided gradually,and arthralgia was relieved.Moreover,the WBC count,ESR and CRP level were decreased to 9.01 × 109/L,50 mm/1 h and 6 mg/L respectively,while the hemoglobin level was increased to 109.7 g/L.After 1-year follow-up,the dosage of glucocorticoids was gradually decreased to 8 mg/d.In addition,his condition was controlled well with no skin lesions and fever,except for occasional arthralgia in the knees and hip.
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Objective To determine the expression of Notch pathway receptors (Notch1 and Notch4) and ligands (Jagged1 and Dll4) in cutaneous malignant melanoma (CMM) tissues,and to preliminarily explore the role of the Notch signaling pathway in the pathogenesis of CMM.Methods Immunohistochemical study was performed to determine the expression pattern and intensity of Notch1,Notch4,Jagged1 and Dll4 in 40 paraffin-embedded CMM specimens and 15 paraffin-embedded pigmented nevus specimens.Statistical analysis was carried out by chi-square test and Spearman rank correlation analysis with the SPSS 21.0 software.Results Notchl was detected in 31 (77.5％) of 40 CMM specimens,as well as in 3 of 15 pigmented nevus specimens,and the positive rates significantly differed between the two groups (x2 =15.281,P ＜ 0.001).However,no significant difference in the expression intensity of Notch1 was observed between 18 in situ melanoma tissues and 22 invasive melanoma tissues (x2 =0.631,P =0.427).In addition,the positive rates of Notch4,Jagged1 and Dll4 were also significantly higher in the CMM group than those in the pigmented nevus group (all P ＜ 0.05),and the expression intensity of Notch4,Jagged1 and Dll4 significantly differed between in situ and invasive melanoma tissues (all P ＜ 0.05).In CMM tissues,the expression of Notch1 was positively correlated with that of Jagged1 (rs =0.350,P =0.027) and Dll4 (rs =0.562,P ＜ 0.001),while the expression of Jaggedl was negatively correlated with that of Dl14 (rs =-0.734,P ＜ 0.001).Conclusion Abnormality of the Notch signaling pathway may be involved in the pathogenesis of melanoma,but further researches are still needed to elucidate the detailed mechanism.
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Objective To construct a eukaryotic expression plasmid carrying the PKCI-1/HINT1 gene,to investigate its expression in A375 melanoma cells after transfection,and to evaluate its effects on apoptosis and autophagy of A375 cells.Methods The PKCI-1/HINT1 gene sequence was amplified by reverse transcription PCR (RT-PCR) with total RNA extracted from A375 cells as the template,then inserted into the eukaryotic expression plasmid PCDNA3.1 (+) to construct a recombinant plasmid,PCDNA3.1 (+)-PKCI-1/HINT1.Some A375 cells were classified into two groups to be transiently transfected with the recombinant plasmid (PCDNA3.1 (+)-PKCI-1/HINT1 group) or the empty plasmid PCDNA3.1 (+) (control group).After additional 48-hour culture,RT-PCR and Western blot analysis were performed to quantify the mRNA and protein expressions of PKCI-1/HINT1 respectively,Hoechst 33342 staining was conducted to detect apoptosis of A375 cells,Western blot analysis to detect the expressions of intracellular caspase-3 and autophagy-associated protein beclin1,and cell autophagy was observed by using the green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 (LC3) labelling method combined with confocal laser scanning microscopy.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of A375 cells at 24,48,72 and 96 hours after transfection.Results Enzyme digestion and sequencing analysis confirmed that the eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed and effectively expressed in the transfected A375 cells.MTT assay showed that PKCI-1/HINT1 could obviously inhibit the proliferation of A375 cells,and the number of live cells was decreased by 17.0％,25.6％ and 29.4％ in the PCDNA3.1 (+)-PKCI-1/HINT1 group at 48,72 and 96 hours,respectively,compared with the control group (all P ＜ 0.05).Hoechest 33258 staining revealed that PKCI-1/HINT1 could promote the formation of apoptotic bodies in A375 cells.Confocal laser scanning microscopy demonstrated that the overexpression of PKCI-1/HINT1 increased GFP-LC3 puncta formation in A375 cells.In addition,Western blot analysis indicated that PKCI-1/HINT1 up-regulated the protein expressions of caspase-3 and beelin1 in A375 cells.Conclusions The eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed,and PKCI-1/HINT1 could be effectively expressed in A375 cells.High-level expression of PKCI-1/HINT1 could suppress cellular proliferation,promote apoptosis,and induce autophagy,of A375 cells.
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Objective To investigate clinical and pathological features of primary cutaneous CD30 + anaplastic large cell lymphoma(PC-ALCL). Methods Clinical and pathological data were collected from 7 patients with PC-ALCL and analyzed retrospectively. Results Of the 7 patients, 6 were male and 1 was female, with an average age of 52 years. PC-ALCL was characterized by solitary (n = 3)or multiple (n = 4)erythematous nodules, lumps and/or plaques with (n = 6)or without (n = 1)ulceration. Systemic involvement was observed in none of the 7 patients. Histopathological examination showed diffuse distribution of tumor cells in the dermis, which were large with rich cytoplasm and atypical nuclei. Mitotic figures were seen. An immunohistochemical study of tumor cells showed positive staining for CD30 and cytotoxic protein, but negative staining for CD20, CD56,anaplastic lymphoma kinase(ALK). Epstein-Barr virus-encoded RNA in situ hybridization was negative. Conclusions PC-ALCL is a rare primary cutaneous low-grade malignant T-cell lymphoma, which can be confirmed by clinical manifestations as well as histopathological and immunohistochemical examinations. It usually has good prognosis with rare systemic involvement and metastasis.
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Objective To investigate clinicopathological features and differential diagnosis of clear cell acan?thoma (CCA). Methods Clinical and pathological data on 10 patients with CCA were retrospectively reviewed. Results CCA clinically manifested as widespread, well?circumscribed, hemispherical dark red to brown papules and nodules with ulcerative, hemorrhagic or desquamative surfaces. Most patients had no subjective symptoms. Nine patients had solitary lesions, and 1 patient had multiple lesions. It frequently occurred in the middle?aged or elderly. Histopathological examination showed thickened prickle cell layer, and the tumor was composed of large clear cells with pale?staining cytoplasm. Characteristic pathological findings were scattered neutrophils and nuclear dust in the epidermis. Periodic acid?Schiff (PAS) staining without diastase was positive in all the 10 patients. Immunohisto?chemical study revealed that tumor cells expressed epithelial membrane antigen (EMA) and keratin, but did not express carcinoembryonic antigen (CEA). Conclusions CCA has no obvious clinical characteristics, and is easily misdiagnosed as melanocytic or vascular tumors. However, CCA has typical histological changes, and histopathological examination is the gold standard for its diagnosis.
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A 38-year-old man complained of multiple papules and nodules in the left retroauricular region for 38 years. On physical examination, there were tens of irregularly sized, skin-colored or pink papules and nodules in the left retroauricular region. Pathological examination showed a large irregularly shaped unilocular cystic space surrounded by a fibrous pseudocapsule in the dermis. The cystic space was lined by a double layer of epithelial cells, including an outer layer of flattened vacuolated myoepithelial cells and an inner layer of cuboidal or columnar secretory cells with eosinophilic cytoplasm. Apocrine secretion was present. Numerous hyperplastic apocrine glands were seen in the deep dermis. A final diagnosis of multiple apocrine hidrocystomas accompanied by apocrine hyperplasia was rendered based on the clinical and pathological presentations.
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Objective To investigate the diagnosis and differential diagnosis of dermatofibrosarcoma protuberans (DFSP). Methods Totally, 50 patients with DFSP visiting the Institute of Dermatology, Chinese Academy of Medical Sciences from 1998 to 2014 were enrolled. The clinical manifestations, histopathological and immunohistochemical features, treatment and prognosis of DFSP were retrospectively reviewed. Results The average age at onset of DFSP was (29.5 ± 15.9)years in the 50 patients, with a mean disease duration of 9.57 years. Skin lesions most frequently occurred on the trunk(n = 33, 66.0%), followed by the extremities, head and neck. DFSP was characterized by atrophic patches or plaques in 13 cases (26.0%), multiple nodules varying in size and arising on atrophic plaques or patches in 30 cases (60.0%), single or multiple nodules arising on normal skin in 7 cases (14.0%). Histologically, the tumor consisted of uniform infiltrative spindle cells arranged in a storiform or cartwheel pattern. In addition, the tumor cells expressed CD34 and vimentin. Twenty patients experienced recurrence at the primary site after resection of skin lesions with a recurrence rate of 43.5%. No distant metastasis or death occurred in these patients. Conclusions DFSP usually has various skin manifestations, is easily misdiagnosed, and can be confirmed based on histopathological and immunohistochemical findings. Local recurrence of DFSP is common, and may occur for many times after surgical excision, but lymphatic and distant metastases are rare.
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Objective To explore the clinicopathologic features of pityriasis lichenoides et varioliformis acuta (PLEVA).Methods A retrospective analysis was performed.Clinical and histological data were collected from 60 patients with PLEVA.The clinicopathologic features of PLEVA were analyzed.Results Among the 60 patients with PLEVA,32 (53.3％) were aged 2-18 years,and 28 (46.7％) aged 19-49 years.Skin lesions were distributed in a diffuse pattern in 50 (83.3％) patients,in a central pattern in 2 (3.3％) patients,and in a peripheral pattern in 8 (13.4％) patients.Nineteen (31.6％) patients had a history of upper respiratory infection.Histopathological examination revealed liquefactive degeneration of basal cells and perivasculitis in the dermis in all the 60 cases,neutrophil abscess formation in the stratum corneum in 26 (43.3％) cases,keratinocyte necrosis in the epidermis in 41 (68.3％) cases,generalized liquefactive degeneration in 30 (50.0％) cases,migration of lymphocytes into the epidermis in 43 (71.6％) cases,Pautrier's microabscess formation in 2 cases,varying degrees of extravasation of erythrocytes into the epidermis in 46 (76.7％) cases,fibrinoid necrosis of blood vessel walls in the dermis in 3 cases.PLEVA progressed into granuloma fungoides in 1 patient.Twenty patients underwent immunohistochemical examination,and 3 of them showed monoclonal hyperplasia of T cells.Conclusions PLEVA has characteristic clinical manifestations,and the combination of pathological and clinical examination is the gold standard for its diagnosis.