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Objective:To evaluate the efficacy of acrivastine alone or in combination with loratadine in the treatment of chronic refractory urticaria.Methods:From March 2017 to December 2018, a multicenter, randomized, controlled clinical study was conducted in 4 centers. Patients with chronic refractory urticaria were randomly divided into two groups, i.e., combined treatment group receiving oral acrivastine capsules 8 mg thrice a day plus oral loratadine tablets 10 mg once a day, and acrivastine alone group receiving oral acrivastine capsules 8 mg thrice a day plus a placebo 10 mg once a day. The course of treatment was 4 weeks. Visits were scheduled at baseline and after 1, 2 and 4 weeks of treatment. At the same time, clinical data were collected, and adverse events were recorded. Symptom scores were evaluated based on degree of itching, number and size of wheals, duration of each attack and number of attacks per week, and symptom score reduce index (SSRI) was used to evaluate the efficacy. Repeated measures analysis of variance and chi-square test were used to evaluate the efficacy and safety.Results:Fifty-three patients in the combined treatment group and 59 in the acrivastine alone group were included in the efficacy analysis. Before treatment, there was no significant difference in symptom score or visual analogue score between the two groups. After 2 weeks of treatment, 19 patients were cured and 10 achieved marked improvement in the combined treatment group, with a response rate of 54.72%; 15 were cured and 6 achieved marked improvement in the acrivastine alone group, with a response rate of 35.59%. After 4 weeks of treatment, 23 patients were cured and 9 achieved marked improvement in the combined treatment group, with a response rate of 60.38%; 20 were cured and 2 achieved marked improvement in the acrivastine alone group, with a response rate of 37.29%. After 2 and 4 weeks of treatment, the response rates were significantly higher in the combined treatment group than in the acrivastine alone group ( χ2 = 4.13, 5.96 respectively, both P < 0.05) . The SSRI significantly differed among different follow-up time points, as well as between the 2 groups ( F = 8.62, 4.38 respectively, both P < 0.05) . Multivariate analysis of variance showed that SSRI was significantly higher in the combined treatment group (0.63 ± 0.05, 0.68 ± 0.05, respectively) than in the acrivastine alone group (0.47 ± 0.05, 0.51 ± 0.05, respectively) after 2 and 4 weeks of treatment (both P < 0.05) . Drug-related adverse reactions, including drowsiness, stomach upsets, headache and liver function abnormality, occurred in 7 patients in the combined treatment group, as well as in 3 in the acrivastine alone group. Conclusion:Acrivastine is safe and effective for the treatment of chronic refractory urticaria, and acrivastine combined with loratadine can markedly improve the efficacy.
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Objective@#To assess the specificity of white dermographism in atopic dermatitis (AD) , and to investigate the relationship between its duration and severity of AD.@*Methods@#From 1st to 30th March 2018, 78 patients with AD (AD group) , 100 patients with non-AD skin diseases (non-AD group) and 100 healthy controls without skin diseases (control group) were enrolled from Department of Dermatology, Wuhan No.1 Hospital. Dermographism test was conducted in each subject, and the subjects′ response and duration of white dermographism were observed. Meanwhile, the severity of skin lesions of the AD patients was evaluated. Statistical analysis was carried out by using Chi-square test, analysis of variance and linear correlation analysis.@*Results@#Of the 78 patients in the AD group, 67 (85.90%) were positive for white dermographism, and the positive rate of white dermographism was significantly higher in the AD group than in the non-AD group (18.00% [18/100], χ2 = 80.97, P<0.017) and control group (5.00% [5/100], χ2 = 119.05, P<0.017) . Additionally, the positive rate of white dermographism was significantly higher in the non-AD group than in the control group (χ2 = 8.30, P<0.017) . The average duration of white dermographism was 1.89 minutes in 30 patients with mild AD, 2.74 minutes in 25 patients with moderate AD, and 4.41 minutes in 12 patients with severe AD. There was a significant difference in the duration of white dermographism among the 3 AD subgroups (F = 64.588, P<0.05) . Moreover, the severity of AD was positively correlated with the duration of white dermographism (r = 0.977, P = 0.136) .@*Conclusion@#White dermographism is a specific clinical manifestation of AD, and its duration is positively correlated with the severity of AD.
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Objective To assess the specificity of white dermographism in atopic dermatitis (AD), and to investigate the relationship between its duration and severity of AD. Methods From 1st to 30th March 2018, 78 patients with AD (AD group), 100 patients with non-AD skin diseases (non-AD group)and 100 healthy controls without skin diseases(control group)were enrolled from Department of Dermatology, Wuhan No.1 Hospital. Dermographism test was conducted in each subject, and the subjects' response and duration of white dermographism were observed. Meanwhile, the severity of skin lesions of the AD patients was evaluated. Statistical analysis was carried out by using Chi-square test, analysis of variance and linear correlation analysis. Results Of the 78 patients in the AD group, 67(85.90％)were positive for white dermographism, and the positive rate of white dermographism was significantly higher in the AD group than in the non-AD group(18.00％[18/100],χ2=80.97, P<0.017)and control group(5.00％[5/100],χ2=119.05, P<0.017). Additionally, the positive rate of white dermographism was significantly higher in the non-AD group than in the control group (χ2 = 8.30, P<0.017). The average duration of white dermographism was 1.89 minutes in 30 patients with mild AD, 2.74 minutes in 25 patients with moderate AD, and 4.41 minutes in 12 patients with severe AD. There was a significant difference in the duration of white dermographism among the 3 AD subgroups(F=64.588, P<0.05). Moreover, the severity of AD was positively correlated with the duration of white dermographism(r=0.977, P=0.136). Conclusion White dermographism is a specific clinical manifestation of AD, and its duration is positively correlated with the severity of AD.
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Objective To evaluate the efficacy and safety of Qingpeng ointment for the treatment of localized eczema in children.Methods A multicenter,randomized,open-labeled,tacrolimus 0.03％ ointment-controlled clinical trial was conducted.A total of 442 children with localized eczema were enrolled into this study,and divided into 2 groups to topically apply Qingpeng ointment (Qingpeng group) and tacrolimus 0.03％ ointment (tacrolimus group) respectively twice a day for 2 weeks.The children were followed up before the treatment and 1,2 weeks after the treatment,and the efficacy and safety were evaluated at the same time.Results Clinical data from 426 children were included in the efficacy analysis.At 1,2 weeks after the treatment,there were no significant differences in the response rate between the Qingpeng group and tacrolimus group (1 week after the treatment:78.92％ vs.81.77％,x2 =0.545,P =0.460;2 weeks after the treatment:98.65％ vs.97.54％,x2 =0.721,P =0.396).However,the pruritus scores at 1,2 weeks after the treatment were both significantly lower in the Qingpeng group than in the tacrolimus group (1 week:F =14.001,P =0.000;2 weeks:F =11.252,P =0.001).At 1 week after the treatment,the incidence rate of adverse reactions was significantly lower in the Qingpeng group than in the tacrolimus group (P ＜ 0.05).Mild itching,burning sensation and other local irritant sensations were the most common adverse reactions.Conclusion Qingpeng ointment is safe and effective for the treatment of localized eczema in children with good tolerability.
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Objective To investigate the mechanism of soluble β-1, 3-D-glucan in G-test positive serum in inhibiting ROS-dependent killing of Candida albicans ( C.albicans ) mediated by neutrophil Dectin-1.Methods The expression and distribution of internalized Dectin-1 and triggered ROS in human neutrophils were detected by using confocal/two-photon laser scanning microscopy upon stimulation with C.albicans (MOI=10) which was pretreated with β-1, 3-D-glucanase (10 U/ml) or not.Abrogation test was used to analyze whether intracellular Dectin-1 was involved in C.albicans-triggered ROS production in human neutrophils.Furthermore, flow cytometry analysis was performed to detect the expression of intracel-lular Dectin-1 and ROS in neutrophils which were pretreated respectively with G-test positive serum at differ-ent dilutions for 60 min and then stimulated with C.albicans for another 60 min at 37℃.Results After stimulated with C.albicans (MOI=10) for 60 min, the expression of Dectin-1 in neutrophils was recruited to the spores of opsonophagocytized C.albicans, and partly co-localized with the triggered ROS production . However, the expression of intracellular Dectin-1 was not observed in neutrophils when stimulated with β-1, 3-D-glucanase pretreated C.albicans for 60 min at 37℃.Abrogation test further showed that C.albicans-trig-gered ROS production in neutrophils was partly and irreversibly inhibited by adding Dectin -1 blocking mAb of 5 μg/ml.In addition , both the triggered expression of intracellular Dectin-1 and ROS production in neu-trophils stimulated with C.albicans ( MOI=10 ) in the presence of G-test positive serum were significantly lower than those of neutrophils stimulated only with C.albicans (LSD-t test, P<0.01).Linear regression a-nalysis suggested that the triggered intracellular Dectin-1 and ROS production in neutrophils upon stimulation with C.albicans were both inhibited by soluble β-1, 3-D-glucan in a dose-dependent manner (Dectin-1,R2=0.702,P<0.01;ROS,R2=0.588,P<0.01 ).Conclusion Taken together, these results demonstrated that the soluble β-1, 3-D-glucan in G-test positive serum may play a role in inhibiting the ROS-dependent killing of C.albicans by interfering with internalized expression of neutrophil Dectin-1.
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Objective To estimate the value of BIOMED-2 primers for the detection of T cell receptor γ (TCR-γ) gene rearrangements in different types of specimens from patients with mycosis fungoides (MF).Methods Totally,15 paraffin-embedded tissue specimens,14 fresh tissue specimens and 18 whole blood specimens were obtained from 28 patients with MF,and subjected to DNA extraction.BIOMED-2 multiplex PCR tubes TCRγ (A+B) were used for the analysis of TCRγgene rearrangements.Data were processed by SPSS 13.0 software,and statistical analysis was done by chi-square test and Fisher's exact probability test.Results TCR-γ gene rearrangements were detected in 3 paraffin-embedded tissue specimens,11 fresh tissue specimens and 12 blood specimens,with significant differences in the detection rate between the three samples (x2 =13.047,P ＜ 0.01).The fresh tissue samples showed a significantly higher detection rate than the paraffin-embedded tissue samples (X2 =12.523,P ＜ 0.01).The detection rate of TCRγgene rearrangements was 3/6 in paraffin-embedded tissue samples collected in 2011,significantly higher than that in the other 9 paraffin-embedded tissue samples collected before 2011 (Fisher's exact probability test,P =0.044),but similar to that in 14 fresh tissue specimens (12/14,Fisher's exact probability test,P =0.044).Decreased detection rate of TCRγ gene rearrangements was observed in blood samples compared with fresh tissue specimens,but no statistical difference was observed between the two types of specimens (x2 =2.358,P ＞ 0.05).Conclusions BIOMED-2 multiplex PCR tubes TCRγ(A+B) are suitable for the detection of clonal rearrangements of TCRγgene in different types of specimens,especially in fresh tissue specimens,from patients with MF.
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Objective To compare the reactive oxygen species (ROS) expression in human neutrophils phagocytizing Sporothrix Schenckii and Candida albicans yeast cells,and to compare the fungicidal activity of human neutrophils against Sporothrix Schenckii and Candida albicans.Methods Human neutrophils were isolated from peripheral blood by using density gradient centrifugation method,and cultured with the presence of the yeast phase of a Sporothrix Schenckii clinical isolate and a standard strain of Candida albicans (ATCC 90028)at a multiplicity of infection of 10 or 1 for 60-210 minutes.Subsequently,flow cytometry with ROS probe (2',7'-dichlorofluorescein diacetate,DCFH-DA) was carried out to for the real time detection of intracellular ROS level,confocal laser scanning microscopy (CLSM) for the observation of ROS distribution.In addition,the fungicidal efficiency of neutrophils against Sporothrix Schenckii and Candida albicans was estimated by the number of colonies after additional culture of neutrophil lysates on brain-heart infusion agar (BHIA) medium.Statistical analysis was done by using univariate analysis of variance and LSD-t test.Results The intracellular ROS level peaked at 60 minutes in neutrophils incubated with Sporothrix Schenckii yeast cells,then decreased rapidly from 60 minutes to 210 minutes.Compared with the neutrophils incubated with Candida albicans yeast cells,those with Sporothrix Schenckii yeast cells showed a higher ROS level (expressed as mean fluorescence intensity) at 60minutes (159.67 ± 11.34 vs.112.22 ± 9.66,P＜ 0.01),but a lower ROS level at 120 minutes (89.01 ± 9.81 vs.110.25 ± 7.28,P＜ 0.05) and 180 minutes (57.63 ± 8.46 vs.109.98 ± 9.00,P＜ 0.01).CLSM revealed that ROS was mainly distributed in neutrophils with phagocytized fungal spores,and especially on the surface of phagocytized spores.Furthermore,the percentage of yeast cells killed by neutrophils was significantly lower for Sporothrix Schenckii than for Candida albicans at 180 minutes (19.21％ ± 3.68％ vs.26.63％ ± 4.97％,P ＜ 0.01).Conclusions Differential expression of intracellular ROS was observed in neutrophils after phagocytosis of Candida albicans and Sporothrix schenckii.Neutrophils exert a stronger fungicidal activity against Sporothrix Schenckii in comparison with Candida albicans,which may be associated with the rapid decrease of ROS level in neutrophils after phagocytosis.
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Objective To investigate the mechanism hy which Dectin-1 mediates the killing of Candida albicans(C.albicans) by human neutrophils in a manner dependent on the production of reactive oxygen species (ROS).Methods After stimulation with FITC-C.albicans at a multiplicity of infection (MOI) of 10 for 30 or 60 min,PE-anti-human Dectin-1 monoclonal antibody (2.5 μg/106 cells) was used to detect the expression of Dectin-1 in human neutrophils by flow cytometry.For Dectin-1 inhibition test and ROS assay,human neutrophils (2×106/ml) were respectively pre-incubated with different concentrations of blocking antibody (0.5,1,2.5 and 5 μg/ml) for 60 min at 4℃,and then with 25 μmol/L 2′,7′-Dichlorofluorescein diacetate for another 20 min at room temperature.Afterwards,under stimulation with live C.albicans at a MOI of 10,the rate of intracellular ROS production over time in blocking and control groups was measured continuously at 10 min intervals for up to 120 min.In addition,localization of Dectin-1 and ROS in human neutrophils was observed by confocal/two-photon laser scanning microscopy after stimulation with live C.albicans.For the detection of candicidal activity,after pre-treatment with different concentrations of Dectin-1 blocking antibody as mentioned above,neutrophils were stimulated with live C.albicans (MOI=1) for 60 min,serial dilutions of cell lysate were plated onto yeast agar,and CFU were enumerated after incubation at 37℃ for 48 h.The candicidal activity was represented as [1-(CFUblocking group/CFUbuffer)] × 100％.Results Under stimulation with FITC-C.albicans at a MOI of 10 for 30 and 60 min,positive percentage of intracellular Dectin-1-expressing neutrophils increased significantly when compared with initial level (0 min,8.32％ ; 30 min,16.82％ ; 60 min,23.88％) (versus 0 min,P＜0.01).However,positive percentage of cell-surface Dectin-1-expressing neutrophils remained almost unchanged after stimulation for 30 and 60 min (versus 0 min,P＞0.05).Upon blockage of Dectin-1,the stimulated ROS generation (R2=0.306,P＜0.01) and candicidal activity (R2=0.251,P＜0.01) of neutrophils were partly and irreversibly inhibited in a dose-dependent manner when compared with control group.In addition,the intracellular Dectin-1 is recruited and co-distributed with ROS on the surface of phagocytized C.albicans as observed by confocal microscopy.Conclusion Taken together,these results demonstrated that an internalized expression pattern of human Dectin-1 might contribute to the ROS-dependent killing of serum-opsonzied C.albicans which was phagocytized by human neutrophils.
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ObjectiveTo study the correlation between the etiology and clinical features of erythroderma.MethodsThe clinical data on 182 patients with erythroderma were retrospectively collected and analyzed.ResultsThe male-to-female ratio was 2.8 ∶ 1 and the average age at onset was 58.6 ± 14.6 years.Of the 182 cases,135 (74.2％) were due to pre-existing dermatoses,14 (7.7％) to drug reaction,8 (4.4％) to malignancies,while 25(13.7％) had no obvious precipitating factors.The most frequent triggering factor was systemic consumption of drugs(52 patients,28.6％ ),and glucocorticosteroid was the most prevalent causative drug.Seventy-six patients were followed up,recurrence was observed in 14 patients but not in 58 patients,and 5 patients died,2 patients with idiopathic erythroderma were finally diagnosed with mycosis fungoides (MF)after multiple skin biopsies.ConclusionsPre-existing dermatoses are the most frequent cause of erythroderma.Idiopathic erythroderma is liable to relapse,possibly associated with malignancies,and should be closely followed up.
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Objective To develop a novel padlock probe and rolling circle amplification(RCA) to detect 5 common cell culture contaminant Mollicute( Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycomplasma orale, and Acholeplasma laidlawii ). Methods "Universal" primers ( SPS1, SPA2 and SPS2, SPA1) were used to amplify the Mollicute 16S-23S rRNA intergenic spacer region.Amplicon was ligate Mollicute specific padlock probe. Probe amplified and monitored using a Corbett RotorGeneTM 6000 machine. Results Five reference Mollicute strains were correctly detected by RCA method.There was no cross-reaction. RCA method can sensitive detect 10 copies templates and show strong positive signal. Sixty-two cell culture specimens were detected. Thirty-seven samples were contained single specie Mollicute and 14 samples contained two Mollicutes. Eleven samples did not contained Mollicute. RCA detection results were concordant with previously species-specific PCR. Conclusion RCA can rapid, sensitive and specific detect contamination Mollicute.
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The histological and immunohistological features of two cases of cutaneous lymphadenoma was studied. A single, erythematous nodule with smooth surface developed on the face of both patients. The lesion slowly progressed. Histology revealed irregular epithelial lobules in the dermis which showed a peripheral palisaded border of basaioid-like cells as well as a center composed of clear cells. Some epithelial lobules and surrounding stroma were infiltrated by numerous small lymphocytes. Immunohistological study showed that the lymphocytic infiltration was predominantly composed of T cells (CD3 positive) along with a small number of B cells (CD20 positive). Within epithelial lobules and surrounded stroma, there were numerous dendritic cells that were positive for S-100 and CDia but negative for cytokeratin 7, cytokeratin 20 or carcino-embryonic antigen. In the center of epithelial lobules in one case, a few cells positive for epithe-lial membrane antigen and CD30 was observed. The diagnosis of cutaneous lymphadenoma was made according to the pathological and immunohistochemical findings, and the infiltration was predominated by CD3-positive lymphocytes in this uncommon epithelial neoplasm.
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Objective To develop a multiple PCR-based reverse line blot hydrization assay (mPCR-RLB)to simutaneously detect several STD pathogens:Neisserria gonorrhoeae,Chlamydia trachomatis,Ureaplasma urealyticum,U.parvum,Mycoplasma genitalium,M,hominis and Trichomonas vaginalis.Methods Seven pairs of biotin-labelled primer were designed and synthesized to target the 16S rRNA-23S rRNA intergenic spacer regions of Neisserria gonorrhoeae,Chlamydia trachomatis,Mycoplasma,and repetitive DNA sequence of Trichomonas to identify and subtype thesc pathogens.DNA was extracted from the referrence strains of seven pathogens and used as templates.mPCR was performed to simutaneously amplify the target regions of these pathogens.Then,the biotin-labelled amplicons were hybridized with membrane-bound specific oligonucleotide probes followed by the detection of bound amplicons with chemiluminescence assay.Serially diluted plasmids containing the target genes of pathogens were amplified with this method to detect its sensitivity.Two-hundred and eleven specimens,including 104 male urethral swabs and 107 female cervical swabs,were collected from the STD clinic of Wuhan First Hospital;mPCR-RLB and single-primer PCR were performed.For specimens with inconsistent results,nested PCR was performed to confirm the results.Results The assay sensitively and specifically identified referrence strains of the tested pathogens.The detection limit of mPCR-RLB was 100 copies for all the pathogens.Of the 211 clinical specimens,2.8%(6/21)were negative for single-primer PCR,but positive for mPCR-RLB,and nested-PCR results were consistent with those of mPCR-RLB.Conclusion mPCR-RLB is a sensitive,specific and rapid method for the detection of STD pathogens from clinical specimens.
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To investigate whether LL-37 and human beta defensin-2 (hBD-2) is related to the patients with psoriasis seldom having skin infections and explore the role of the two peptides and CCR6 (the receptor of hBD-2) in the pathogenesis of psoriasis, the expression levels of mRNA of LL-37, hBD-2, and CCR6 in skin lesions of patients with psoriasis vulgaris were detected by using RT-PCR. The results showed that the mRNA expression levels of the two peptides and CCR6 in psoriatic lesions all increased compared with the normal skin (P<0.001). It was suggested that up-regulated expression of LL-37 and hBD-2 might be the main reason that result in the the skin of patients with psoriasis being seldom infected, and the two peptides and CCR6 might play crucial roles in the pathogenesis of psoriasis.
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Adult , Female , Humans , Male , Middle Aged , Antimicrobial Cationic Peptides , Genetics , Psoriasis , Genetics , Metabolism , RNA, Messenger , Genetics , Receptors, CCR6 , Receptors, Chemokine , Genetics , Up-Regulation , beta-Defensins , GeneticsABSTRACT
Objectives To investigate the expression of B lymphocyte stimulator (BLyS) and its receptor BAFF-R(a receptor of B-cell activator factor, belonging to the TNF family) in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE), and to explore their clinical significance. Methods The patients were separated into active group (n = 28) and inactive group (n = 24) according to the SLE disease activity index (SLEDAI). The mRNA and protein expression of BLyS and its receptor BAFF-R were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blot in PBMCs from the patients and 21 healthy volunteers. The relationship between the expression of BLyS and BAFF-R and SLEDAI was analyzed. Results In the case of the expression of mRNA and protein of BLyS and BAFF-R, the patients had a higher level than the healthy controls (P 0.05). Conclusions These findings indicate that BLyS and its receptor BAFF-R might be involved in the pathogenesis of SLE. Also, BLyS expression level might be a new parameter for the evaluation of SLE disease activity and therapeutic effect.