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Objective:To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) in the diagnosis of bullous pemphigoid (BP) .Methods:A single-center clinical retrospective study was conducted. Totally, 163 patients with newly diagnosed BP were collected from Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2013 to January 2019, so were 404 controls, including 161 with pemphigus, 67 with eczema, 26 with drug eruption, 23 with erythema multiforme, 18 with prurigo nodularis, etc. Blood samples were collected before the treatment, and IIF-SSS, BP180 NC16A enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) assay were performed to evaluate the value of IIF-SSS in the diagnosis of BP. Measurement data were compared by using t test and Mann-Whitney test, and enumeration data were compared by using chi-square test and Fisher′s exact test or McNemar test. Results:The number of cases positive for IIF-SSS, BP180 NC16A ELISA and DIF assay was 160, 153 and 127 respectively in the BP group, and 0, 18 and 26 respectively in the control group. The sensitivities of IIF-SSS, BP180 NC16A ELISA and DIF assay for the diagnosis of BP were 98.15%, 93.86% and 77.91% respectively, and their specificities were 100%, 95.54% and 93.56% respectively. There was strong consistency in the diagnosis of BP between IIF-SSS and DIF (Kappa coefficient= 0.767, P < 0.001) . Conclusion:IIF-SSS has relatively high sensitivity and specificity for the diagnosis of BP, and can serve as a routine method for diagnosing BP.
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Objective:To optimize indirect immunofluorescence on salt-split skin (IIF-SSS), and to evaluate its performance in detection of bullous pemphigoid (BP) antibodies.Methods:Normal human foreskin and non-foreskin skin tissues were used to prepare salt-split substrates under 3 different experimental conditions: traditional group rotated at 4 ℃ for 48 - 72 hours, low-temperature immersion group soaked at 4 ℃ for 48 - 72 hours, room-temperature immersion group soaked at 25 ℃ (range: 23 - 27 ℃) for 24 hours. Serum samples were obtained from 20 patients with bullous pemphigoid (BP) in Hospital of Dermatology, Chinese Academy of Medical Sciences between August 2019 and August 2020, and subjected to IIF on the intact skin or salt-split substrates by using a multiple dilution method. Paired-sample t test was used for comparisons of means between two paired samples. Results:No dermal-epidermal separation was observed in the substrates prepared in the low-temperature immersion group at 48 - 72 hours, while dermal-epidermal separation occurred in the lower lamina lucida of the foreskin and non-foreskin substrates in the room-temperature immersion group and the traditional group. For the 20 patients with BP, the reciprocal end-point titers ( M[ Q1, Q3]) detected with the salt-split non-foreskin skin and salt-split foreskin in the room-temperature immersion group, and with the salt-split non-foreskin skin in the traditional group were 5 120 (2 560, 17 920), 1 280 (640, 2 560), 1 280 (640, 2 560), respectively. Moreover, 19 (95%) patients with BP showed that the reciprocal end-point titers detected with the substrates in the room-temperature immersion group were 1 - 5 times those in the traditional group ( t = 8.04, P<0.001), suggesting that the performance of salt-split skin in the room-temperature immersion group was superior to that in the traditional group in the detection of BP antibodies; however, there was no significant difference in the reciprocal end-point titers of BP antibodies between the salt-split foreskin in the room-temperature immersion group and salt-split non-foreskin skin in the traditional group ( t<0.001, P>0.05). The reciprocal end-point titers in 20 BP sera detected by conventional IIF on the intact non-foreskin skin and foreskin were 320 (160, 640) and 480 (160, 1 120), respectively; the reciprocal end-point titers detected by IIF on the salt-split foreskin and non-foreskin skin in the room-temperature immersion group, as well as on the salt-split non-foreskin skin in the traditional group, were all consistent with or 1 - 7 times higher than those detected by conventional IIF ( t = 6.47, 14.83, 5.26, respectively, all P<0.001) . Conclusion:The soaking method at room temperature 25 ℃ (23 - 27 ℃) for preparing salt-split substrates has advantages of short duration and simple procedure, and the sensitivity of IIF-SSS using the substrates prepared by this method is equal or superior to the traditional salt-split method for detecting BP antibodies.
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Objective:To investigate the correlation of related antibody titers in serum of patients with pemphigus vulgaris (PV) with disease severity and activity.Methods:A total of 24 patients with active PV were collected, who firstly visited Hospital for Skin Diseases, Chinese Academy of Medical Sciences from 2012 to 2015. Pemphigus disease area index (PDAI) was evaluated in the patients with PV at active and stable stages, and serum samples were collected. Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of pathogenic anti-desmoglein (Dsg) conformational epitope antibodies, total anti-Dsg antibodies and anti-acetylcholine receptor (AChR) antibody in serum samples. Measurement data were compared by using t test, enumeration data were compared by using Fisher′s exact test, and correlations were analyzed by using Pearson correlation analysis. Results:Among the patients with active PV, there was no significant difference between the anti-Dsg1 antibody titers (611.4 ± 136.8) and anti-Dsg1 conformational epitope antibody titers (585.5 ± 134.7, t = 0.13, P = 0.89) , but the anti-Dsg3 antibody titers (708.6 ± 130.7) were significantly higher than the anti-Dsg3 conformational epitope antibody titers (297.2 ± 54.4, t = 2.90, P < 0.01) . In addition, both the anti-Dsg1 antibody titers and anti-Dsg1 conformational epitope antibody titers were positively correlated with PDAI scores in the patients with active PV (both r = 0.54, P < 0.01) ; PDAI scores were not correlated with the anti-Dsg3 antibody titers ( r = 0.11, P = 0.62) , but positively correlated with the anti-Dsg3 conformational epitope antibody titers ( r = 0.53, P < 0.01) . Among the 20 patients with stable PV, the serum titers of anti-Dsg1 antibodies and anti-Dsg1 conformational epitope antibodies significantly decreased compared with those at their first visit; anti-Dsg3 antibody titers significantly decreased in only 7 patients, and 13 patients still had high titers of anti-Dsg3 antibodies, including 6 with declined anti-Dsg3 conformational epitope antibody titers, and 5 converted from anti-AChR antibody-positive to anti-AChR antibody-negative. Conclusions:Both the anti-Dsg1 antibody and anti-Dsg1 conformational epitope antibody titers can reflect the disease activity of PV. The disease activity was not consistent with anti-Dsg3 antibody titers in some patients, and the anti-Dsg3 conformational epitope antibody or anti-AChR antibody may facilitate evaluating the disease activity of PV.
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Pregabalin can reduce the release of multiple neurotransmitters by acting on the voltagegated calcium channel of the nervous system.It is currently widely used in a variety of diseases,including neuropathic pain,generalized anxiety disorder,epilepsy and so on.In dermatology department,pregabalin also has a therapeutic effect on postherpetic neuralgia,prurigo nodularis,uremic pruritus,nerve-related pruritus and mentally relevant pruritus.
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Vaccine is an important measure to prevent diseases,and plays a great role in preventing various infectious diseases.Vaccine can not only produce a beneficial "immune response",but also induce adverse reactions or even adverse damage,some of which manifest as skin lesions.Clinical manifestations of vaccine-associated cutaneous adverse reactions are various,and their pathogenesis is complex.This review mainly elaborates the current status of vaccine-associated cutaneous adverse reactions,which may help to understand the pathogenesis and different clinical manifestations of vaccine adverse reactions.
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Objective To prepare human epidermal extracts by thermal separation,and to evaluate the value of epidermal extract-based Western blot analysis in the diagnosis of bullous pemphigoid (BP).Methods Human epidermal extracts were prepared by thermal separation from circumcised foreskins of healthy males.Serum samples were obtained from 22 inpatients with BP and 25 inpatients without BP in Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and August 2017.These serum samples were subjected to Western blot analysis with epidermal extracts as substrates,as well as to BP180-NC16A enzyme-linked immunosorbent assay (ELISA).Statistical analysis was carried out using chi-square test and Fisher's exact test with the SPSS22.0 software.Results The sensitivities of epidermal extract-based Western blot analysis and BP 180-NC16A ELISA in the diagnosis of BP were 86.36％ (95 ％ CI:64.03％-96.41％) and 95.45％ (95％ CI:75.11％-99.76％) respectively (~ =1.10,P =0.294),and the specificities were 100％ (95％ CI:83.42％-100％) and 92％ (95％ CI:75.11％-99.76％) respectively (x2 =20.8,P =0.149).Epidermal extract-based Western blot analysis in the 22 patients with BP showed a protein band with relative molecular mass (RMM) of 230 000 in 4 patients,a protein band with RMM of 180 000 in 18,a protein band with RMM of 120 000 in 1,and a protein band with RMM of 97 000 in 1.The BP180-NC16A ELISA showed that the antibody titers were more than 50 U/ml in the BP patients with protein bands of RMM of 180 000.Conclusions The epidermal extract-based Western blot analysis mainly showed the protein band with RMM of 180 000 in the patients with BP.The sensitivity of the epidermal extract-based Western blot analysis was lower than that of the BP180-NC16A ELISA,and the epidermal extract-based Western blot analysis tends to be negative when the titer of the autoantibody is low.
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Objective To investigate associations of anti-desmoglein (Dsg1 and Dsg3) antibodies detected by enzyme-linked immunosorbent assay (ELISA) with clinical phenotypes and disease activity in pemphigus patients,and to explore their change patterns.Methods A total of 111 patients with pemphigus were enrolled from Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and January 2018.ELISA was performed to detect serum levels of anti-Dsg1 and anti-Dsg3 antibodies in these patients with different clinical types of pemphigus at different stages,including onset stage,control stage (no new erythema or vesicles occurred in the last 2 or more weeks,and primary lesions began to regress),maintenance stage (the condition had been stable for ≥ 1 month,and treatment was maintained with a low dose of glucocorticoids [prednisone equivalent of ＜ 15 mg/d]),and recurrence stage,and the change patterns of serum levels of anti-Dsg1 and anti-Dsg3 antibodies were analyzed.Statistical analysis was carried out with SPSS 22 software by using oneway analysis of variance for the comparison among groups,and least significant difference (LSD)-t test for multiple comparisons.Results At the disease onset stage,control stage,maintenance stage and recurrence stage,92,53,33,and 9 patients respectively completed the detection.Among the 92 patients with initial onset of pemphigus,the positive rates of anti-Dsg1 and anti-Dsg3 antibodies were 100％ and 2.77％ respectively in 36 patients with pemphigus foliaceus,20％ and 80％ respectively in 10 with mucosaldominant pemphigus vulgaris,and 97.82％,95.65％ respectively in 46 with mucocutaneous pemphigus vulgaris.The serum levels of anti-Dsg1 antibodies in the patients with pemphigus foliaceus significantly differed among the disease onset stage,control stage,maintenance stage and recurrence stage (137.43 ±77.74,13.94 ± 14.81,21.50 ± 58.33,121.13 ± 86.89 U/ml,respectively),the serum levels of anti-Dsg3 antibodies in the patients with mucosal-dominant pemphigus vulgaris also significantly differed among the above clinical stages (125.61 ± 94.81,34.5 ± 16.26,0.6,258 U/ml,respectively),and the serum levels of anti-Dsg1 and anti-Dsg3 antibodies in patients with mucocutaneous pemphigus vulgaris both significantly differed among the above clinical stages(anti-Dsg1 antibody:115.39 ± 70.62,15.74 ± 25.10,3.62 ± 12.09,78.60 ± 92.25 U/ml,respectively;anti-Dsg3 antibody:137.98 ± 81.25,58.14 ± 63.46,29.26 ± 64.70,136.9 ± 101.47 U/ml,respectively).Additionally,the serum levels of anti-Dsg1 antibodies in the patients with pemphigus foliaceus,as well as the serum levels of anti-Dsg3 antibodies in the patients with mucosaldominant pemphigus vulgaris and those with mucocutaneous pemphigus vulgaris,were both significantly lower at the disease control stage and maintenance stage than at the disease onset stage and recurrence stage (all P ＜ 0.05).During the treatment,epitope spreading occurred in 2 patients,and high-titer anti-Dsg antibodies were observed in 4 patients at the stable stage.Conclusion Anti-Dsg antibody spectrum is associated with clinical phenotypes of pemphigus,and its serum levels measured by ELISA can be applied to disease activity monitoring and evaluation of therapeutic efficacy.
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Skin diseases manifesting as desquamative gingivitis (DG) can be divided into recurrent DG-and chronic DG-related skin diseases,including oral lichen planus,mucosal pemphigoid,pemphigus vulgaris and so on.A thorough medical history,detailed oral and histopathological examinations and serum immunological tests can be helpful for correct diagnosis of DG-related skin diseases.The treatment of DG-related skin diseases includes topical and systemic therapies.It is necessary to individualize treatment protocols due to treatment response.During the treatment of DG,oral hygiene should be strengthened,secondary fungal and bacterial infections should be avoided,and attention should be paid to the protection of oral cavity and periodontal tissues.
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Objective To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) and bullous pemphigoid 180 N C 16a enzyme-linked immunosorbent assay (BP 180 N C 16a-ELISA) in the diagnosis of bullous pemphigoid (BP).Methods Serum samples were collected from 174 BP patients and 129 controls,who were enrolled from Institute of Dermatology of Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and August 2017,and subjected to IIF-SSS and BP180 NC16a-ELISA.Direct immunofluorescence (DIF) test was performed in 25 cases of BP,and its sensitivity for the diagnosis of BP was compared with that of IIF-SSS and BP180 NC16a-ELISA.Results The sensitivities for IIF-SSS and BP180 NC16a-ELISA were 93.67％ and 96.55％ respectively,and the specificities for IIF-SSS and BP180 NC16a-ELISA were 100％ and 96.12％ respectively.IIF-SSS was weakly correlated with BP180 NC16a-ELISA with a correlation coefficient of 0.147.There was no significant difference in the sensitivity between the serological diagnostic methods (IIF-SSS and BP180 NC 16a-ELISA) and DIF.Conclusion Serological diagnostic methods show high specificity and sensitivity in the diagnosis of BP,and are worthy of clinical promotion and application.