Epitope spreading in autoimmune bullous diseases / 中华皮肤科杂志

Epitope spreading often occurs in patients with autoimmune bullous diseases (ABDs), resulting in exposure of more antigenic epitopes, aggravation or transformation of pre-existing diseases, or concurrence of other diseases. With the increase in the immunological evidence for epitope spreading, more and more scholars have realized that epitope spreading plays an important role in the development of ABDs. This review introduces the phenomenon of epitope spreading in ABDs from 4 aspects, including the concurrence of or transformation between different types of pemphigoid, different types of pemphigus, pemphigus and pemphigoid, as well as between ABDs and other skin diseases.

Correlation between autoimmune bullous diseases and psoriasis / 中华皮肤科杂志

There are overlaps in the pathogenesis of autoimmune bullous diseases (AIBDs) and psoriasis. In recent years, there have been many reports on correlations between psoriasis and AIBDs. Bullous pemphigoid is the most common AIBD associated with psoriasis, followed by pemphigus vulgaris, pemphigus erythematosus and linear IgA bullous dermatosis. AIBDs occur after the onset of psoriasis in most cases, but simultaneously with or before the onset of psoriasis in some patients. This review summarizes correlations between AIBDs and psoriasis, as well as possible mechanisms.

A case report of dermatitis herpetiformis with positive gluten-sensitive intestinal disease autoantibodies / 中华检验医学杂志

A middle-aged male patient initially appeared scattered erythema with pruritus all over the body without obvious cause. According to the skin manifestations of the patient, combined with pathological diagnosis, direct immunofluorescence examination, and different serum autoantibody spectrum, the diagnosis and differential diagnosis of the patient was made by clinicians. The diagnosis of dermatitis herpetiformis was confirmed by the use of autoantibodies in the absence of any apparent history of pasta discomfort. With targeted treatment, the patient′s symptoms and laboratory indicators improved significantly.

Nonbullous pemphigoid / 中华皮肤科杂志

Nonbullous pemphigoid (NBP) , which is related to bullous pemphigoid, has various clinical manifestations, and is frequently accompanied by itching. Typical clinical manifestations of bullous pemphigoid (BP) , such as tense blisters or bullae, are absent in NBP cases. It is easy to misdiagnose. Histopathological findings are not specific, and its diagnosis should be confirmed by direct immunofluo-rescence, indirect immunofluorescence or salt-split indirect immunofluorescence. NBP may develop into BP in some cases, and the prognosis of NBP is better than that of BP. However, delayed diagnosis usually leads to a relatively high dosage of drugs for disease control, and a high rate of adverse reactions.

Action mechanisms of thymic stromal lymphopoietin in various skin diseases / 中华皮肤科杂志

Thymic stromal lymphopoietin (TSLP) , a cytokine similar to interleukin-7 (IL-7) , can promote the differentiation and proliferation of a variety of cells, promote the secretion of Th2 cytokines by these cells, and plays an important role in the immune system. In recent years, abnormal expression of TSLP has been found in many skin diseases, and its level is also related to the severity of some skin diseases, suggesting that TSLP may be a potential target for the treatment of various skin diseases. This review summarizes recent research progress in the role of TSLP in the occurrence and development of various skin diseases, including inflammatory diseases, immune diseases, genetic diseases and tumors, and provides a basis and some ideas for the diagnosis and treatment of related diseases.

Detection of eosinophil cationic protein in sera and blister fluids of patients with bullous pemphigoid / 中华皮肤科杂志

Objective:To investigate the relationship of eosinophil cationic protein (ECP) levels in sera and blister fluids with bullous pemphigoid (BP) .Methods:From January 2012 to October 2019, 40 patients with newly diagnosed BP and 40 healthy individuals were enrolled from Department of Dermatology, Peking Union Medical College Hospital, and serum ECP levels were detected by enzyme-linked immunosorbent assay. Meanwhile, another 33 patients with newly diagnosed BP and 41 patients with non-autoimmune bullous diseases were enrolled, and the ECP level was detected in blister fluids by enzyme-linked immunosorbent assay. Pathological sections of skin lesions of 1 patient with BP and 1 with contact dermatitis were subjected to immunohistochemical staining for ECP. Normally distributed data were compared between 2 groups by using t test or t′ test, while enumeration data were compared by using chi-square test. Pearson′s correlation coefficient was used to analyze the relationship between serum levels of ECP and proportions of peripheral blood eosinophils in patients with BP. Results:The serum level of ECP was significantly higher in the BP group (116.9 ± 19.3 ng/L) than in the healthy control group (93.3 ± 15.9 ng/L, t = 5.96, P<0.001) , and the blister fluid level of ECP was also significantly higher in the BP group (665.8 ± 189.0 ng/L) than in the non-autoimmune bullous disease group (547.5 ± 240.6 ng/L, t = 2.31, P = 0.02) . Immunohistochemical study showed more brown-yellow particles in the cytoplasm of ECP-positive cells in the BP group than in the contact dermatitis group. There was no significant correlation between the serum level of ECP and proportion of peripheral blood eosinophils in the BP patients ( r = -0.15, P = 0.35) . Conclusion:The levels of ECP in the sera and blister fluids markedly increased in the patients with BP, and blister fluid levels of ECP were much higher than serum levels of ECP, suggesting that ECP may be involved in the occurrence of BP.

There is no correlation between the serum level of interleukin-21 and bullous pemphigoid / 中华皮肤科杂志

Objective To investigate the correlations of the serum level of interleukin-21 (IL-21) with anti-BP 180 antibodies,peripheral eosinophil level,bullous pemphigoid disease area index (BPDAI)in patients with bullous pemphigoid (BP).Methods A total of 31 patients with bullous pemphigoid and 31 healthy controls were enrolled into this study.Blood samples were collected from all the subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum level of IL-21 and serum titer of anti-BP180 antibodies.BPDAI was assessed among 13 patients,and the correlation of the IL-21 level with anti-BP180 antibody titers,eosinophil count and BPDAI were analyzed.Results No significant difference in the serum level of IL-21 was observed between the patients (M [P25-P75]:255.28 [156.19-361.59] ng/L) and healthy controls (150.47 [72.4-268.73] ng/L,P =0.14).There was no correlation between the serum level of IL-21 and serum titer of anti-BP180 antibodies (82.98 [26.82-132.92] U/ml,rs =0.18,P =0.332).Among the 27 BP patients who underwent blood cell count,no correlation was found between the serum levels of IL-21 and whole-blood eosinophil counts (rs =0.104,P =0.606).Among the 13 BP patients who underwent assessment of BPDAI,there was no correlation between the serum levels of IL-21 and BPDAI scores (rs =0.233,P =0.464).Conclusion The IL-21 level is uncorrelated with anti-BP 180 antibody level,peripheral eosinophil level or BPDAI in patients with BP.

Prognostic accuracy of the SCORTEN scoring system in patients with toxic epidermal necrolysis or Stevens-Johnson syndrome / 中华皮肤科杂志

Objective To evaluate the prognostic accuracy of the score of toxic epidermal necrolysis (SCORTEN) scoring system in patients with toxic epidermal necrolysis (TEN) or Stevens-Johnson syndrome (SJS).Methods Clinical data were collected from 39 patients with SJS/TEN hospitalized in Peking Union Medical College Hospital during April 1992 and March 2014,and retrospectively analyzed.Among the 39 patients,13 had died,and the other 26 patients,who were matched to the dead patients in a ratio of 2:1 for age,all had a definite diagnosis and were discharged with improved conditions.The SCORTEN scoring system was used to evaluate the 39 patients with SJS/TEN and calculate expected mortality.The expected mortality and actual mortality were compared between different groups stratified by age in the 39 patients.The receiver operating characteristic (ROC) curve was drawn to assess the prognostic accuracy of the SCORTEN scoring system.Results According to the SCORTEN scoring system,15 out of the 39 patients scored 1 point,14 scored 2 points,6 scored 3 points,and 4 scored 4 points.The total number of expected deaths was 6.808,while that of actual deaths was 13.There was no significant difference between the expected mortality and actual mortality in every SCORTEN score-based group.The area under curve (AUC) was 0.832 8,indicating a good predictive ability of the SCORTEN scoring system.Conclusion The SCORTEN scoring system can predict mortality in TEN/SJS patients at early stage.

Clinical measures of bullous pemphigoid / 中华皮肤科杂志

Bullous pemphigoid (BP) is a group of autoimmune blistering skin diseases.There are special measures for the assessment of disease severity and therapeutic response in autoimmune bullous skin diseases.Common scoring indexes include autoimmune bullous skin disorder intensity score (ABSIS),bullous pemphigoid disease area index (BPDAI),and so on.ABSIS has been validated,and is widely applied in evaluation of pemphigus vulgaris and other autoimmune blistering diseases.BPDAI is still in the process of verification,and has been applied in only a few clinical trials.The evaluation of diseases by ABSIS and BPDAI is both based on a global assessment of skin and mucosal lesions,and degree of subjective pruritus.

Combined therapy of psoralen plus ultraviolet A followed by narrow band ultraviolet B photochemotherapy for early stage mycosis fungoides / 中华肿瘤杂志

<p><b>OBJECTIVE</b>Only a few clinical reports in the treatment of early mycosis fungoides (MF)(IA, IB, IIA stage) are available in the literature. The purpose of this study was to compare the efficacy and safety of narrow-band UVB and psoralen plus ultraviolet A (PUVA) photochemoterapy in 24 patients with early-stage MF, and explore a new approach for the treatment of early mycosis fungoides.</p><p><b>METHODS</b>A total of 24 identified early mycosis fungoides patients received PUVA, NB-UVB and a combined therapy of PUVA followed by NB-UVB (n = 9/6/9) irradiation. A retrospective study was carried out to analyze the sex, age of onset, TNM stage, treatment, and duration of treatment, and times of treatment, duration of maintenance treatment, effective and recurrence in these patients. The data were analyzed using SPSS 17.0 and a two-sided test at the α = 0.05 level of significance was conducted.</p><p><b>RESULTS</b>Of the 24 patients studied, the average treatment was 104.5 (95% CI, 75.71-133.29) times. The average duration of treatment was 12.88 (95% CI, 9.90-15.85) months. The average maintenance treatment time was 11.08 (95% CI, 2.13-20.04) months. The effective rate (CR+PR) of PUVA treatment was 88.9%, recurrence rate was 11.1% (n = 9). In the NB-UVB treatment group, the effective rate was 100.0%, and the recurrence rate was 33.3% (n = 6). In the PUVA followed by NB-UVB (combination therapy) treatment group, the effective rate was 77.8% and the recurrence rate was 55.6% (n = 9). There were no significant differences among the three groups in terms of number of treatments, treatment duration, maintenance treatment duration, effective rate and recurrence rate (P > 0.05).</p><p><b>CONCLUSIONS</b>PUVA and NB-UVB are effective and safe in the targeted therapy of early stage mycosis fungoides. The combined therapy of PUVA followed by NB-UVB can reduce the total PUVA dose and risk of developing skin cancer.</p>

Development of a method to detect anti-BP180NC16A IgG subclasses and the significance of antiBP180NC16A IgG in bullous pemphigoid / 中华皮肤科杂志

ObjectiveTo develop an assay to quantitatively detect anti-BP180NC16A IgG subclasses and to assess the significance of anti-BP180NC16A lgG in bullous pemphigoid (BP).MethodsThe Glutathione S-transferase(GST)-BPI80NC16A fusion protein was expressed in E.coli system and purified by affinity chromatography.An improved enzyme-linked immunoabsorbent assay (ELISA) was developed and used to detect anti-BP180NCI6A IgG subclasses in serum samples from 10 patients with BP,5 patients with pemphigoid gestationis (PG),1 patient with linear IgA bullous dermatosis (LIBD) and 2 patients with pemphigus.Results The optimal condition for the ELISA was determined by cross assay as follows:the concentration of GSTBP180NC16A fusion protein for coating,500 μg/L; the condition for coating,4 ℃ for 12 hours; the dilution ratio of sera and secondary antibody,1∶ 100 and 1 ∶ 2000 respectively; the condition for incubation,37 ℃ for 1 hour;,the condition for the enzyme-substrate reaction,37 ℃ for 20 minutes.Of the 10 patients with BP,all were positive for anti-BP180NC16A IgG1,9 for IgG2,5 for IgG3,and 9 for IgG4.Anti-BP180NC16A IgG was undetected in any of the serum samples from 2 patients with pemphigus vulgaris or 1 patient with adult LIBD.All the 5 sera from patients with PG; were positive for all the anti-BP180NC16A IgG subclasses,which were predominated by IgGl and IgG3.ConclusionThe developed ELISA is a highly specific and reproducible semiquantitative method for the detection of anti-BP180NCI6A IgG subclasses in patients with BP and PG.

Purification and identification of anti-BP180 NC16A antibodies from the sera of patients with bullous pemphigoid or herpes gestationis / 中华皮肤科杂志

Objective To develop a method to purify and identify anti-BP180 NC16A antibodies from the sera of patients with bullous pemphigoid (BP) or herpes gestationis.Methods The GST/NC16A fusion protein was expressed in a prokaryotic expression vector pGEX-2TBP180NC16A,and then crosslinked to glutathione sepharose beads.Anti-BP180 NC16A antibodies were isolated from the sera of 3 patients with BP and 2 patients with herpes gestationis by affinity chromatography,and analyzed by immunofluorescence,Western blot and enzyme linked immunosorbent assay (ELISA).Results The GST/NC16A fusion protein with a relative molecular mass of 37 000 was successfully expressed by the prokaryotic vector pGEX-2TBP180NC16A.Purified anti-BP180 NC16A antibodies were obtained from the sera of patients by the affinity chromatography,and ELISA revealed that the concentration of anti-BP180 NC16A was 2.4 mg/ml.The purified antibody could bind to the basement membrane zone of human skin,suggesting a strong biological activity of the antibodies.Western blot showed a single band corresponding to the expected molecular mass for anti-BP180 NC16A antibodies,indicating a high purity of the isolated antibodies.Conclusion The anti-BP180 NC16A antibodies purified by microbead-based affinity chromatography from the sera of patients with BP or herpes gestationis are highly active and specific.

Expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis / 中华皮肤科杂志

Objective To measure the expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis (EV), and to explore the relationship between cellular immune abnormality and EV pathogenesis. Methods Immunohistochemistry was used to measure the expressions of IL-10, IL-23 and CD86 in tissue samples from 10 patients with EV and 10 normal human controls. Results Three cytokines were observed in all the samples of EV, with the expression score ranging from 3 to 6 and expression intensity from moderate to high. However, of the control specimens, only 1 was positive for IL-10 with the expression score being 3, and expression intensity being moderate. Conclusion The pathogenesis of epidermodysplasia verruciformis may be correlated with the expression abnormality of some cytokines secreted by keratinocytes.

Relationship of TAP-1 allele gene polymorphism to recurrent condyloma acuminatum / 中华皮肤科杂志

Objective To investigate the relationship of polymorphisms at codons 333 and 637 of TAP-1 allele gene to recurrent condyloma acuminatum (CA) in a Chinese population. Methods Amplificatory refraction mutation system-PCR (ARMS-PCR) was performed to detect TAP1 polymorphic residues at codon 333 in 88 patients with recurrent CA and 81 age- and sampling date-matched controls and at codon 637 in 60 patients with recurrent CA and 60 age- and sampling date-matched controls. Results The frequencies of AA,GG and AG genotypes at codon 333 of TAP-1 gene were 86.36%, 0, 13.64%, respectively, in patients with recurrent CA, 79.01%, 0 and 20.99%, respectively, in the controls, and no significant difference was observed between the two groups (χ2 = 1.604, P ＞ 0.05). However, a significant difference was observed in the frequencies of AA, GG and AG genotypes at codon 637 of TAP-1 gene between the patients and controls (3.33% vs 10.0%, 95.00% vs 60.00%, 1.67% vs 30.00%, χ2 = 21.551, P ＜ 0.01 ). Conclusions The recurrence of CA may be associated with the polymorphism at codon 637, but not with that at codon 333, of TAP-1 gene.

The Mutations and Sequencing of HPV16E6E7Gene of Human Papillomavirus from Patients with HPV Infection in Beijing / 中华皮肤科杂志

Objective To investigate the gene sequence and mutations of human papillomavirus(HPV)type16E6E7in patients with HPV infection in Beijing.Methods Sample DNA was extracted from lesions in patients with HPV infection.HPV types were identified by polymerase chain reaction(PCR).E6E7gene,isolated from samples infected with HPV16only,was cloned into plasmid pGEM-3zf and sequenced.Results The recombinant plasmid pGEM/16E6E7was constructed successfully.The whole HPV E6E7gene was776bp in length which was equal to that of the standard strain.Three nucleotides exchanges,i.e.,p60PROE6,p96GLUE6,p565SERE7,were found in E6E7gene.Conclusion The data suggest that there are nucleotide differences of HPV E6E7gene between HPV obtained from Beijing and that of standard sequence.

Efficacy of Q-switched Alexandrite laser in seborrheic keratosis / 中华医学美学美容杂志

Objective Seborrheic keratosis are very common dermatosis occurred in the fourth to fifth decade. Recently, some encouraging results in the treatment of melain-related lesions, such as Ota`s nevus, tatoos, cafe spots, have been obtained using the Q-switched Alexandrite laser, but no reports were found in treating seborrheic keratosis. Our purpose was to assess the clinical outcomes induced by the Q-switched Alexandrite laser at 752nm. Methods Total 119 cases of seborrheic keratoses , aged from 31-76years, were treated with the Q-switched Alexandrite laser (model HT-10, Cynosure Laser Corp, USA. wavelength 752nm, pulse width 45-75ns). The energy was adjusted to 7.1-8.5J/cm 2 according to the charateristics of the lesions. The interval of two treatment was 1.5-3 months. No special care was needed. Results Sixty-seven of the 119 patients were evaluated after treatment (men 15 cases, women 52 cases, aged from 31～71 years, and course from 1 to 21 years). All the patients were treated by 1-3 times. The number of cases treated by 1, 2, and 3 times was 50 (74.6 %), 14 (20.9 %) and 3 (4.5 %), respectively. The degree of clearing was: complete clearing (17 cases, 25.4 %), significant improvement (25 cases, 37.3 %), moderate improvement (17 cases,25.4 %) and no change or mild improvement (8 cases,11.9%). The total rate of improment was 62.7%, while the improvement rate after 1 treatment was 58%. The efficacy was correlated with the area of the lesions. No side effects, such as hyperpigmentation and scars, were found. Conclusions This clinical data demonstrate the Q-switched Alexandrite laser is a safe and effective method for treatment of seborrheic keratosis and deserves to be used widely.

PCR-induced Modification of C Terminus of HPV-16 E7 and Expression of Mutational E7 in Eukaryotic Cells / 中华皮肤科杂志

Objective To induce the mutation of HPV-16 E7 in two zinc-binding motifs near the C terminus by polymerase chain reaction (PCR) and evaluate the effect of this mutation on the antigen-specific immunity of HPV-16 E7. Methods HPV-16 E7 fragment was amplified by PCR and cloned into pGEM-3zf vector. Two site mutations at 58 and 91 animo acid sites in the open reading frame of HPV-16 E7 were induced by PCR, and then the molecular clones of HPV-16 E7 wild type (pcDNA3.1/E7) and mutant (pcDNA3.1/ME7) were successfully reconstructed. Western blot and immunofluorescence were used to detect the expression of E7 protein. Results Intracellular fluorescence signals were observed in the cells transfected with pcDNA3.1/E7 and pcDNA3.1/ME7 24 hours after transfection, but the signals in the cells transfected with pcDNA3.1/ME7 disappeared 48 hours after tansfection. Twenty-four and 48 hours after transfection with pcDNA3.1/ME7, E7 protein was not detected by Western blot. Conclusions The stability of HPV-16 E7 protein is reduced by mutations (C58G, C91G) near two zinc-binding motifs. It is suggested that the zinc-binding motifs near the C terminus of HPV-16 E7 may be important for maintaining the stability of E7 protein.

Detecting and Typing of HPV DNA in Tissues of Some External Genital Proliferative Diseases / 中华皮肤科杂志

Objective To study the possible relationship between human papillomaviruse (HPV) infection and external genital proliferative diseases. Methods HPV DNA was detected using polymerase chain reaction (PCR) with a consensus primer and typed by using restriction fragment length polymorphism (RFLP) method in 151 paraffin-embedded biopsy specimens from seven kinds of external genital epidermal proliferative lesions. Results In 30 cases of condyloma acuminatum, HPV DNA was positive for all cases, among which HPV6 and HPV11 accounted for 60% and 26.7%, respectively. In 40 cases of bowenoid papulosis, 5 cases of genital Bowen′s disease and 6 cases of erythroplasia of Queyrat, the positive rates of HPV DNA were 55%, 100% and 33.3%, respectively, and the predominant type was HPV16. In 18 cases of external genital invasive squamous cell carcinoma HPV DNA positive rate was 27.8%, and HPV16 was detected in all of the 5 HPV-positive cases. HPV DNA was negative in 32 cases of leukoplakia and 20 cases of extramammary Paget′s disease. Conclusions Condyloma acuminatum is mainly caused by HPV6 infection, followed by HPV11; HPV16 infection is closely associated with the pathogenesis of bowenoid papulosis and external genital squmous cell carcinoma including Bowen′s disease and erythroplasia of Queyrat. In external genital invasive squamous cell carcinoma, HPV16 infection may be one of the multiple carcinogenic factors. No clear relationship is found between HPV infection and leukoplakia or extramammary Paget′s disease.