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Objective:To construct "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " based on the Delphi method, and to provide a methodological basis for consensus construction.Methods:After systematic search and evaluation of the literature related to chronic inducible urticaria, the first draft of "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " was written, and a questionnaire was designed for expert consultation. A representative sample of 25 experts was selected to conduct two rounds of correspondence consultation via electronic questionnaire in strict accordance with the Delphi method, and the content of the consensus was revised and improved according to the consultation results.Results:The response rates in the two rounds of questionnaire consultation were both 100%, and the expert authority coefficient was 0.92 ± 0.09. In the first round of consultation, the coefficients of variation (CV values) of 9 items were greater than 20%, and the mean agreement degree of 3 items was less than 7 points; in the second round of consultation, the CV values of all items were less than 15%, the agreement degree of the above 3 items whose mean agreement degree was less than 7 points in the first round of consultation all rose to over 7 points, and the median agreement degree of all items was greater than or equal to 8 points. Reliability analysis of the two rounds of questionnaire results showed that the Cronbach α coefficient and standardized Cronbach α coefficient were both greater than 0.9; the P values in the agreement tests by using Kendall′s coefficient of concordance for the two rounds of questionnaire results were both less than 0.001, and the Kendall′s coefficients of concordance were 0.170 and 0.219 in the first and second rounds of questionnaire consultation, respectively. Conclusion:The Delphi method-based "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " is highly representative, authoritative and reliable; this study also provides a methodological reference for the formulation and research of consensus.
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Objective:To explore the association rules of personality traits and health-promoting lifestyle in patients with primary angle closure glaucoma, so as to provide advice for the synthetical treatment.Methods:From July to November 2021, a total of 117 primary angle closure glaucoma patients(acute patients n=89, chronic patients n=28) in ophthalmology department of five hospitals in Nanjing were investigated with type A behavior pattern scale, health-promoting lifestyle scale Ⅱ and general information questionnaire.Based on Weka 3.8.5, algorithm of Apriori was used to mine its association relationship. Results:(1) The total scores of type A behavior pattern scale for patients with acute and chronic types of primary angle closure glaucoma were (32.48±6.43) and (27.54±6.49) respectively.The total scores of health-promoting lifestyle scale Ⅱ were (101.69±11.83) and (97.79±7.78) respectively.(2) There were positive associations among patients with acute primary angle closure glaucoma, type A/A-personality (including impatience and hostility) and health-promoting lifestyle (including stress management disorder, interpersonal relationship management disorder, well sense of health responsibility and adequate dietary nutrition intake)(all support>0.1, confidence >0.6, lift >1.0). And patients with chronic primary angle closure glaucoma were associated with B/B-personality (including patience and mild), health-promoting lifestyle (including stress management disorder, interpersonal relationship management disorder, well sense of health responsibility and adequate dietary nutrition intake)(all support>0.1, confidence >0.6, lift >1.0).Conclusion:Primary angle closure glaucoma is strongly related with personality traits and health-promoting lifestyle.Its synthetical treatment plan should take both physical and mental measures, and classified health management for patients with different disease types.
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Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare but severe diseases. This study aimed to validate the predictive ability of risk models in patients with SJS/TEN and propose possible refinement in China. Patients in the Department of Dermatology of Huashan Hospital from January 2008 to January 2019 were included. Results showed that the severity-of-illness score for TEN (SCORTEN) had a good discrimination (area under the receiver operating characteristic curve (AUC), 0.78), and it was superior to auxiliary score (AS) and ABCD-10, which indicates age, bicarbonate level, cancer, dialysis, and 10% involved body surface area (AUC, 0.69 and 0.68, respectively). The calibration of SCORTEN (Hosmer-Lemeshow goodness-of-fit test, P = 0.69) was also better than that of AS (P = 0.25) and ABCD-10 (P = 0.55). SCORTEN and ABCD-10 were similar (Brier score (BS), 0.04 and 0.04) in terms of accuracy of predictions. In addition, the imaging appearance of pulmonary consolidation on computed tomography was associated with high mortality. Refined models were formed using the variables and this imaging appearance. The refined AS and ABCD-10 models were similar in discrimination compared with the original SCORTEN (0.74 vs. 0.78, P = 0.23; 0.74 vs. 0.78, P = 0.30, respectively). Therefore, SCORTEN showed good discrimination performance, calibration, and accuracy, and refined AS or ABCD-10 model may be an option when SCORTEN variables are not available.
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Humans , Cohort Studies , Retrospective Studies , Severity of Illness Index , Stevens-Johnson Syndrome/diagnostic imaging , TomographyABSTRACT
Objective@#To evaluate the in vitro effect of tacrolimus on the expression and function of protease-activated receptor 2 (PAR-2) in cultured human keratinocytes.@*Methods@#After 24-hour co-culture of human keratinocytes with 10-9 - 10-5 mol/L tacrolimus, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed to determine the mRNA expression of PAR-2, immunofluorescence (IF) staining and Western blot analysis were performed to determine the protein expression of PAR-2 in the keratinocytes, and the fluorescent calcium probe fluo-4 was used to evaluate the effect of tacrolimus at different concentrations on the intracellular calcium concentration after the activation of PAR-2 in the keratinocytes. The group treated without tacrolimus served as control group. One-way analysis of variance was used to compare the PAR-2 expression and calcium concentration in the human keratinocytes in different groups, and least significant difference (LSD) -t test was carried out for multiple comparisons.@*Results@#PAR-2 was expressed on both the membrane and cytoplasm of keratinocytes. After 24-hour co-culture of keratinocytes with 10-9 - 10-5 mol/L tacrolimus, the PAR-2 mRNA expression significantly decreased in these cells compared with the control group (all P < 0.05) , and was negatively correlated with the tacrolimus concentration (r=-0.962, P = 0.009) . IF staining and Western blot analysis showed that the PAR-2 protein expression was significantly lower in the 10-5- and 10-6-mol/L tacrolimus groups than in the control group (both P < 0.05) , and decreased to a certain extent in the 10-7-mol/L tacrolimus group (IF staining: P < 0.05; Western blot analysis: P > 0.05) . No significant difference in the PAR-2 protein expression was observed between the 10-8- or 10-9-mol/L tacrolimus group and the control group (both P > 0.05) . After 24-hour co-culture, the peak concentration of intracellular calcium after PAR-2 activation was significantly lower in the 10-5-, 10-6- and 10-7-mol/L tacrolimus groups (peak absorbance: 1 463 ± 283, 1 455 ± 270, 1 423 ± 291 respectively) than in the control group (1 602 ± 407; t = 2.582, 2.821, 2.923, P = 0.032, 0.022, 0.019, respectively) , while there was no significant difference between the 10-8- or 10-9-mol/L tacrolimus group (1 649 ± 379, 1 633 ± 415 respectively) and the control group (t = 0.846, 0.462, P = 0.422, 0.657, respectively) .@*Conclusion@#Tacrolimus can inhibit PAR-2 expression and suppress calcium mobilization induced by a PAR-2 agonist in keratinocytes.
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Objective To evaluate the in vitro effect of tacrolimus on the expression and function of protease-activated receptor 2(PAR-2)in cultured human keratinocytes. Methods After 24-hour co-culture of human keratinocytes with 10-9-10-5 mol/L tacrolimus, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR)was performed to determine the mRNA expression of PAR-2, immunofluorescence(IF)staining and Western blot analysis were performed to determine the protein expression of PAR-2 in the keratinocytes, and the fluorescent calcium probe fluo-4 was used to evaluate the effect of tacrolimus at different concentrations on the intracellular calcium concentration after the activation of PAR-2 in the keratinocytes. The group treated without tacrolimus served as control group. One-way analysis of variance was used to compare the PAR-2 expression and calcium concentration in the human keratinocytes in different groups, and least significant difference(LSD)-t test was carried out for multiple comparisons. Results PAR-2 was expressed on both the membrane and cytoplasm of keratinocytes. After 24-hour co-culture of keratinocytes with 10 - 9- 10 - 5 mol/L tacrolimus, the PAR-2 mRNA expression significantly decreased in these cells compared with the control group(all P < 0.05), and was negatively correlated with the tacrolimus concentration(r=-0.962, P=0.009). IF staining and Western blot analysis showed that the PAR-2 protein expression was significantly lower in the 10-5- and 10-6-mol/L tacrolimus groups than in the control group (both P < 0.05), and decreased to a certain extent in the 10-7-mol/L tacrolimus group(IF staining:P<0.05;Western blot analysis:P>0.05). No significant difference in thePAR-2 protein expression was observed between the 10-8-or 10-9-mol/L tacrolimus group and the control group(both P>0.05). After 24-hour co-culture, the peak concentration of intracellular calcium after PAR-2 activation was significantly lower in the 10-5-, 10-6- and 10-7-mol/L tacrolimus groups (peak absorbance:1463 ± 283, 1455 ± 270, 1423 ± 291 respectively)than in the control group(1602 ± 407;t=2.582, 2.821, 2.923, P=0.032, 0.022, 0.019, respectively), while there was no significant difference between the 10-8-or 10-9-mol/L tacrolimus group(1649 ± 379, 1633 ± 415 respectively)and the control group(t=0.846, 0.462, P=0.422, 0.657, respectively). Conclusion Tacrolimus can inhibit PAR-2 expression and suppress calcium mobilization induced by a PAR-2 agonist in keratinocytes.
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Objective To evaluate the inductive effect of interferon-γ(IFN-γ) combined with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on programmed necrosis of the human immortalized keratinocyte cell line HaCaT,and to explore its mechanisms.Methods In vitro cultured HaCaT cells were divided into several groups:negative control group receiving no treatment,IFN-γ group treated with 50 μg/L IFN-γ,TRAIL group treated with 4 μg/L TRAIL,and cytokine combination group treated with 50 μg/L IFN-γ and 4 μg/L TRAIL or zVAD combination group pretreated with 40 μmo/L zVAD for 1 hour followed by the treatment with 50 μg/L IFN-γand 4 μg/L TRAIL.After 48-hour treatment,the morphology of HaCaT cells were observed under a light microscope,methyl-thiazolyl-tetrazolium assay was performed to evaluate the inhibitory effect of the treatment on the proliferation of HaCaT cells,and double staining flow cytometry to detect the necrosis of HaCaT cells.Meanwhile,real-time fluorescence-based quantitative PCR (qPCR) was conducted to determine the mRNA expression of receptor interaction protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL),Western blot analysis to determine the expression of RIP1,RIP3,MLKL proteins and their phosphorylated forms (pRIP1,pRIP3,pMLKL),immunofluorescent staining to observe the distribution of pRIP3 and pMLKL in HaCaT cells,and the level of reactive oxygen species (ROS) in HaCaT cells in the above groups was detected by the fluorescence probe DCFH-DA.Statistical analysis was carried out with SPSS 22 software by using one-way analysis of variance (ANOVA) for comparing indices among different groups,and least significant difference (LSD)-t test for multiple comparisons.Results After 48-hour treatment,HaCaT cells in the cytokine combination group and zVAD combination group showed necrosis-like morphologic features.Methyl-thiazolyl-tetrazoliumassay revealed significant differences in the survival rate of HaCaT cells among the IFN-γgroup,TRAIL group,cytokine combination group,zVAD combination group and negative control group (73.16％ ± 5.71％,81.46％ ± 4.68％,72.18％ ± 2.93％,69.67％ ± 3.24％ and 100％,respectively;F =24.34,P ＜ 0.001).The necrosis rate of HaCaT cells was notably higher in the cytokine combination group and zVAD combination group (9.86％ ± 1.31％,10.33％ ± 2.16％,respectively) than in the negative control group (5.26％ ± 0.91％,t =4.61,5.07,respectively,both P ＜ 0.05).qPCR revealed that the mRNA expression of RIP3 and MLKL significantly increased in the cytokine combination group and zVAD combination group compared with the negative control group (tRIP3 =0.99,1.84,tMLKL =1.51,2.17,respectively,all P ＜ 0.05).Western blot analysis suggested that the protein expression of RIP1,RIP3,MLKL,pRIP1,pRIP3 and pMLKL significantly increased in the cytokine combination group compared with the negative control group (all P ＜ 0.05),and the zVAD combination group showed significantly decreased caspase 8 expression and increased expression of the above proteins compared with the cytokine combination group.Fluorescence microscopy showed that enhanced green dot-like or clump-like fluorescent spots (representing pRIP3) could be observed in the cytoplasm,and red fluorescent spots (representing pMLKL) could be seen on the cell membrane in the cytokine combination group.The average fluorescence intensity of ROS was significantly higher in the cytokine combination group than in the negative control group (t =702.00,P ＜ 0.05).Conclusion IFN-γcombined with TRAIL can induce the programmed necrosis of HaCaT cells with increased level of ROS.
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Objective@#To explore the clinical value of endoscopic interventional therapy for locally recurrent primary lung adenoid cystic carcinoma (ACC).@*Methods@#The clinical data of 42 patients with locally recurrent ACC were retrospectively analyzed, and the differences of tracheal and bronchial diameter, airway scoring grade and airway obstruction degree before and after treatment were compared among three treatment methods: bronchoscopic interventional therapy + palliative radiotherapy, interventional therapy alone, and non-interventional therapy. Log rank test and Cox proportional risk model multi-factor analysis were used to determine the prognostic factors of ACC patients with local recurrence, and the long-term effect of bronchoscopic interventional therapy on ACC with local recurrence was determined.@*Results@#The median overall survival of 42 patients was 59 months and 5-year survival rate was 54.2%.Univariate analysis showed that vascularized cancer, pleural invasion, pulmonary atelectasis, incisal margin, microscopic classification, tumor diameter, initial TNM stage, ki-67 index, and treatment after local recurrence were associated with long-term survival of ACC patients with local recurrence (all P<0.05). Cox multivariate regression analysis showed that margin status (RR=0.272, P=0.011), tumor diameter (RR=2.586, P=0.005), initial TNM staging (RR=0.369, P=0.035), ki-67 index (RR=3.569, P<0.001), and treatment methods after local recurrence (RR=0.126, P<0.001) were independent factors influencing the prognosis of ACC patients with local recurrence. After three months of treatment, the tracheal bronchus diameters, rating of shortness of breath, and degree of airway obstruction were all improved significantly (all P<0.05), both in the interventional therapy + palliative radiotherapy group [(14.5±2.8 mm, 0.86±0.45, (14.50±10.67)%, respectively], and the interventional therapy alone group [(13.7±2.3) mm, 0.97±0.25, (15.38±12.02)%, respectively]. Meanwhile, the difference before and after non-interventional therapy was not statistically significant (all P>0.05). 5-year overall survival rates were 55.8%, 46.6% and 42.6% for patients undergoing interventional therapy+ palliative radiotherapy, interventional therapy alone, and non-interventional therapy after recurrence, with statistically significant differences (P=0.015). Patients underwent bronchial endoscopic interventional therapy and palliative radiotherapy had the best efficacy of treatment.@*Conclusion@#Endoscopic interventional therapy plus palliative radiotherapy is an effective local palliative treatment for locally recurrent ACC patients, which can rapidly relieve airway stenosis, improve the quality of life of patients and prolong the survival time of patients.
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Objective To explore the in vitro culture methods for oriented differentiation of peritoneal cells and bone marrow cells into high-purity mast cells,and to identify the function of these mast cells.Methods Peritoneal cells and bone marrow cells were isolated from the peritoneal cavity lavages and femur of C57BL/6 mice,and cultured with both interleukin-3 (IL-3) and stem cell factor for 2 and 4 weeks respectively.Light microscopy was performed to observe the morphology of these cells,toluidine blue staining to identify the degree of maturity of these mast cells,and flow cytometry to measure the expression of cell surface markers C D 117 and FceR Ⅰ α.After the stimulation with compound 48/80 at different concentrations,the degranulation rate of mast cells was counted under the microscope,and β-hexosaminidase release rate was measured by spectrophotometry.Results After 2-or 4-week culture,the mouse peritoneal and bone marrow cells all manifested as refractive suspension cells of uniform size.Toluidine blue staining showed violaceous metachromatic granules in the cytoplasm of the two kinds of cells.The proportions of CD117 or FcεR Ⅰ α single-positive peritoneal and bone marrow-derived mast cells were all more than 95％,and the proportions of CD117/FcεR Ⅰ α double-positive peritoneal and bone marrow-derived mast cells were 97.68％ ± 0.80％ and 96.12％ ± 0.76％ respectively.The degranulation rates of mast cells in the 100-and 1 000-mg/L compound 48/80 groups significantly differed from those in the blank control group (all P ＜ 0.01).Compared with the blank control group,the β-hexosaminidase release rates significantly increased in bone marrow-derived mast cells in the 100-mg/L compound 48/80 group and peritoneal mast cells in the 10-and 100-mg/L compound 48/80 groups (P ＜ 0.01 or 0.05).Conclusion IL-3 and stem cell factor can co-induce the directed differentiation and proliferation of mouse bone marrow stem cells and peritoneal cells,so as to harvest highnuritv mature degranulated mast cells,and lay a foundation for subsequent cell biology research.
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Chronic urticaria is a very common allergic disease in the outpatient.It has complicated pathogenesis and there is no conclusion up to now.The activation of the mast cells and the release of the histamine are the key point of the chronic urticaria onset.In recent years,the studies have demonstrated that the autoimmunity and the chronic infection are the main cause of the disease.There are other factors including dysfunction of the coagulation,deficiency of the vitamin D,pseudoallergen,psychic factor and genetic mechanism.This article will make a review of the pathogenesis of the chronic urticaria from the aspects mentioned above.
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Objective:To analyse the whole DNA methylation status , expression of DNMT1 and DNA methylation status of DNMT1 promoter from patients with systemic lupus erythematosus ( SLE) .To assess the negative feedback effection of DNA methylation on the expression of DNMT1 in the patients with SLE.Methods:The whole DNA methylation status in T cells from 34 SLE patients and 23 healthy controls was assessed by the specific monoclonal antibodies to 5-methylcytosine (5-mc) and the DNA methylation status of DNMT1 promoter was assessed by the Bisulfite Conversion Kit and sequencing.Real time RT-PCR was applied to analyse DNMT 1 mRNA levels in T cells from the patients and controls.Results: SLE patients had siginificantly whole DNA hypomethylation than controls(P<0.05),and the whole DNA methylation was negative correlated with the SLE-DAI.There was no difference in levels of DNA methylation of DNMT1 promoter between patients and controls ,also no correlation between the levels of DNA methylation of DNMT 1 promoter and DNMT1 mRNA.Conclusion:There was no negative feedback effection of DNA methylation on the expression of DNMT 1 in the patients with SLE.The mechanism of lower expression of DNMT 1 in patients with SLE should be studied further.
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Objective To investigate DNA methylation status as well as signal transducer and activator of transcription 3 (STAT3) gene expression in peripheral blood T-lymphocytes from patients with systemic lupus erythematosus (SLE),and to explore their relationship.Methods Peripheral blood samples were obtained from 34 patients with SLE and 23 healthy controls followed by the isolation of T-lymphocytes.Flow cytometry was performed to evaluate DNA methylation status using antibodies against 5-methylcytosine (5-me),reverse transcription (RT)-PCR to detect the mRNA expression of STAT3 in T-lymphocytes.Results Compared with healthy controls,17 patients with active SLE showed significantly decreased DNA methylation levels (8.50 ± 1.42 vs.11.31 ± 1.34,P ＜ 0.01),but increased STAT3 mRNA expression levels (1.34 ± 0.32 vs.1.02 ± 0.28,P ＜ 0.01).However,there were no significant differences between 17 patients with stable SLE and healthy controls in DNA methylation levels (11.30 ± 1.34 vs.11.31 ±1.34,P ＞ 0.05) or STAT3 mRNA expression levels (1.01 ± 0.27 vs.1.02 ± 0.28,P ＞ 0.05).Patients with active SLE also showed significantly reduced DNA methylation levels,but enhanced STAT3 mRNA expression compared with those with stable SLE (both P ＜ 0.01).As Pearson correlation analysis showed,SLE disease activity index (SLE-DAI) was negatively correlated with DNA methylation levels in patients with SLE (r =-0.78,P ＜ 0.01),but positively correlated with STAT3 mRNA expression (r =0.50,P ＜ 0.01),and no significant correlation was observed between STAT3 mRNA expression and DNA methylation levels (r =-0.13,P ＞ 0.05).Conclusion The aberrant expression of STAT3 in peripheral blood T-lymphocytes may be related to disease activity in SLE,but unrelated to DNA methylation levels.
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Objective To measure the levels of plasma D-dimer,activated coagulation factor Ⅶ (FⅦa) and activated clotting factor Ⅻ (FⅫa) in patients with chronic urticaria (CU),and to investigate their relationship with the occurrence of CU.Methods Venous blood samples were collected from 50 patients with CU and 50 healthy human controls.Dry-column immune scattering chromatography was performed to detect the plasma level of D-dimer,and enzyme-linked immunosorbent assay (ELISA) to measure the levels of FⅦa and FⅫa.In addition,autologous plasma skin test (APST) was conducted in 43 patients with CU,and autologous serum skin test (ASST) in 41 patients with CU.A correlation analysis was carried out between the above three parameters and disease severity as well as between the results of APST and ASST and plasma level of D-dimer.Results The levels of plasma D-dimer and F Ⅶa were significantly higher in patients with CU than in healthy human controls (both P ＜ 0.05),while no significant difference was found in FⅫa level between the two groups (P ＞ 0.05).Moreover,the degree of increase in D-dimer plasma level was positively correlated with disease severity in the patients with CU.The plasma level of D-dimer was significantly higher in APST-positive patients than in APST-negative patients (P ＜ 0.05),but not significantly different between ASST-positive and-negative patients (P ＞ 0.05).Conclusions Coagulation mechanism,especially the extrinsic coagulation pathway,is related to the occurrence of CU.Studies on coagulation mechanism are beneficial to the evaluation of severity,and clinical treatment,of CU.
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Objective To investigate the expressions of miR-17 and miR-20a in CD4+ T cells from patients with systemic lupus erythematosus (SLE) and their correlations with disease activity.Methods CD4+ T cells were isolated from the venous blood of 30 patients with SLE and 18 healthy human controls.RNA was extracted from the CD4+T cells,and real-time fluorescence-based quantitative PCR (qPCR) was performed to determine the expression levels of miR-17 and miR-20a.The intergroup comparison of miR-17 and miR-20a expression levels was done using t test and Mann-Whitney test,and the correlations of miR-17 and miR-20a expressions with clinical parameters were assessed using Pearson or Spearman correlation coefficients.Results The patients with SLE showed increased expression levels (2-△c~) of miR-17 and miR-20a compared with the healthy controls (0.24 ± 0.08 vs.0.15 ± 0.06 for miR-17,0.19 ± 0.10 vs.0.12 ± 0.09 for miR-20a,both P ＜ 0.05).The miR-17 and miR-20a expression levels were also significantly higher in patients with active SLE than in those with inactive SLE and the healthy controls (0.27 ± 0.07 vs.0.20 ± 0.05 and 0.16 ± 0.08 for miR-17,0.22 ± 0.10 vs.0.15-0.07 and 0.13 ± 0.09 for miR-20a,all P ＜ 0.05),but similar between the patients with inactive SLE and healthy controls (both P ＞ 0.05).The expression levels of both miR-17 and miR-20a were positively correlated with SLE disease activity index (SLEDAI)(r =0.45,0.38,respectively,both P ＜ 0.05) and anti-dsDNA antibody level (r =0.54,0.46,respectively,both P ＜0.05),but negatively correlated with serum C3 levels (r =-0.43,-0.42,respectively,both P ＜ 0.05).Condusions The expressions of miR-17 and miR-20a are increased in CD4+ T cells from patients with SLE,and correlated with the disease activity in SLE.
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Arabidopsis BOTRYTIS-INDUCED KINASE1 (BIK1) is a receptor-like cytoplasmic kinase acting early in multiple signaling pathways important for plant growth and innate immunity. It is known to form a signaling complex with a cell-surface receptor FLS2 and a co-receptor kinase BAK1 to transduce signals upon perception of pathogen-associated molecular patterns (PAMPs). Although site-specific phosphorylation is speculated to mediate the activation and function of BIK1, few studies have been devoted to complete profiling of BIK1 phosphorylation residues. Here, we identified nineteen in vitro autophosphorylation sites of BIK1 including three phosphotyrosine sites, thereby proving BIK1 is a dual-specificity kinase for the first time. The kinase activity of BIK1 substitution mutants were explicitly assessed using quantitative mass spectrometry (MS). Thr-237, Thr-242 and Tyr-250 were found to most significantly affect BIK1 activity in autophosphorylation and phosphorylation of BAK1 in vitro. A structural model of BIK1 was built to further illustrate the molecular functions of specific phosphorylation residues. We also mapped new sites of FLS2 phosphorylation by BIK1, which are different from those by BAK1. These in vitro results could provide new hypotheses for more in-depth in vivo studies leading to deeper understanding of how phosphorylation contributes to BIK1 activation and mediates downstream signaling specificity.
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Amino Acids , Chemistry , Arabidopsis , Arabidopsis Proteins , Chemistry , Genetics , Gene Expression Regulation, Plant , Immunity, Innate , Mutation , Phosphorylation , Protein Serine-Threonine Kinases , Chemistry , Genetics , Signal Transduction , Threonine , GeneticsABSTRACT
Objective To evaluate skin barrier function in children with atopic dermatitis (AD) as well as healthy children from two communities in Shanghai and to assess the relationship between skin barrier function and AD severity.Methods Totally,169 children with AD and 142 healthy children aged 3-12 years were recruited from two communities (Changning Xining community and Jiading Juyuan community) in Shanghai,China.Transepidermal water loss (TEWL) and stratum corneum hydration were measured in normal appearing nonlesional skin at four body sites (dorsal and volar forearm,cheek and anterior shin) of the patients,as well as in normal skin at the same sites of the controls.AD severity was evaluated by using the severity scoring of atopic dermatitis (SCORAD) index.Results Compared with the healthy children,the patients with AD showed higher TEWL value at all of the four body sites (all P ＜ 0.05),but lower water content in stratum corneum at dorsal forearm and anterior shin (both P ＜ 0.05).In patients with AD,the SCORAD index was positively correlated with mean TEWL value,but negatively correlated with the mean water content in stratum corneum.Conclusion Skin barrier function may serve as an index for evaluating the severity of AD.
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Objective To assess the clinical features and associated factors of interstitial lung disease (ILD) in patients with dermatomyositis (DM).Methods Clinical data were retrospectively analyzed on 206 patients with DM collected at the Department of Dermatology,Huashan Hospital,Fudan University in the past 6 years.Chi-square test and t test were performed for statistical analysis.Results The prevalence of ILD was 49.03％ in the 206 patients with DM.Heliotrope rash on the upper eyelids,Gottron's sign (papules),arthralgia,and cough were correlated with the incidence of ILD in DM patients (all P ＜ 0.05),and of these factors,the prevalence of artharalgia and cough were positively correlated with the incidence of ILD,while the presence of Gottron's papules was negatively correlated.The patients with DM and ILD showed a higher prevalence of abnormal serum levels of lactate dehydrogenase (LDH),hydroxybutyrate dehydrogenase (HBDH) and anti-Jo-1 antibodies,as well as with a poorer pulmonary function,compared with those suffering from DM only (all P ＜0.05).Characteristic imaging findings on computed tomography (CT) scan in patients with DM and ILD included linear opacity (57.4％),high-density patchy opacity (31.7％),reticular opacity (16.8％) and even ground glass-like opacity (13.8％),usually at the bottom or apex of the lungs.Conclusions In patients with DM,the prevalence of artharalgia and cough is positively correlated,whereas the presence of Gottron's papules is negatively correlated,with the incidence of ILD.Characteristic imaging findings on CT scan in patients with DM and ILD are linear opacity,high-density patchy opacity,reticular opacity and ground glass-like opacity at the bottom and apex of lungs.
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Objective To evaluate the efficacy and safety of Qingpeng ointment in the treatment of eczema.Methods A multi-center,randomized,double-blind and placebo-controlled clinical trial was conducted.A total of 246 patients with eczema were randomly assigned with a ratio of 2∶1 to the treatment group and control group to topically apply Qingpeng ointment and placebo respectively twice daily for 3 weeks.Total symptom scores were calculated for the patients at the baseline,on week 1,2 and 3 during the treatment according to the individual scores for pruritus,lesions including erythema,papules,papulovesicles or vesicles,desquamation,crusting,infiltration and lichenification.The occurrence of adverse events was recorded.Results Totally,228 patients completed the trial,including 154 patients in the treatment group and 74 patients in the control group.After 3 weeks of treatment,a statistical difference was observed in the response rate (85.71％ vs.41.89％,Z=47.16,P＜ 0.01) and cure rate (31.82％ vs.12.16％,Z=12.30,P＜ 0.01) between the treatment and control group.There was no significant difference in the incidence of adverse events between the two groups (2.48％ vs.2.56％,x2 =0,P ＞ 0.05).Conclusion Qingpeng ointment displays a promising efficacy for the treatment of mild to moderate eczema with a rapid onset and high safety.
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Objective To investigate the effects of 5-azacytidine and estradiol on the methylation of CpG motifs and expression of DNA methyltransferasel (DNMT1) mRNA in patients with systemic lupus erythematosus (SLE) and normal controls.Methods Peripheral blood lymphocytes isolated from 12 patients with SLE and 11 normal human controls were stimulated with phytohaemagglutinin for one day followed by additional 3-day culture with or without the presence of 5-azacytidine of 1 μmol/L or estradiol of 30μg/L respectively.Then.the methylation of CpG motifs was detected by flow cytometry using anti-5-methylcytosine antibody,and DNMT1 mRNA expression by real time reverse transcription-PCR Results After treatment with 5-azacytidine,a decrease wag observed in the methylation of CpG motifs, but not in the expression of DNMT1 mRNA in peripheral lymphocytes from patients with SLE (1=18.60,P<0.01;t=1.56.P>0.05) and in those from the normal controls (t=5.63,P<0.01;t=2.17,P>0.05) compared with untreated lymphocytes.Nevertheless,there were no significant changes in the methylation of CpG motifs or expression of DNMT1 mRNA in lymphocytes from patients with SLE (t=1.53,0.93,respectively,both P>0.05) and normal controls (t=1.93,0.11,respectively,both P>0.05) after the treatment with estradiol.Conclusions The methylation of CpG motifs is suppressed efficiently by 5-azacytidine,and the suppression is unlikely to be associated with the decrease of DNMT1 mRNA.Estradiol has no significant impact on the methylation of CpG motifs and expression of DNMT1 mRNA in lymphocytes.
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Objective To explore the mechanism underlying the induction of systemic lupus erythematosus (SLE) by estrogen, hydralazine and ultraviolet irradiation. Methods Peripheral blood mononuclear cells (PBMCs) were harvested from 10 patients with SLE and 9 normal human controls, and cultured with or without the intervention with estrogen, hydralazine or ultraviolet irradiation. The DNA methyltransferase-1 (DNMT1) activity of PBMCs was quantified by using DNMT activity/inhibition assay kit. Results No statistical difference was observed in DNMT1 activity between patients with SLE and normal controls (0.36 ± 0.24 vs 0.46 ± 0.17, P ＞ 0.05). A significant decrease was noted in DNMT1 activity in PBMCs from patients with SLE after intervention with estrogen (0.32 ± 0.18 vs 0.46 ± 0.17, t = 1.725, P ＜ 0.05), hydralazine (0.33 ±0.13 vs 0.46 ± 0.17, t = 1.739, P ＜ 0.05) and ultraviolet irradiation (0.30 ± 0.14 vs 0.46 ± 0.17, t = 1.739,P ＜ 0.05 ) compared with that from normal human controls. The treatment with hydralazine also induced an attenuation of DNMT1 activity in PBMCs from normal human controls (0.38 ± 0.12 vs 0.46 ± 0.17, P＜ 0.05).Conclusion Estrogen, hydralazine and ultraviolet irradiation can inhibit the DNMT1 activity of SLE patients,indicating that they may induce the initiation of SLE by altering the activity of DNMT1.
ABSTRACT
Aim To explore the effect of microRNAs on the differentiation of 3T3-L1 adipocytes and the expression of adipo-related gene-fatty acid binding protein during the adipocyte differentiation.Methods adipo-related microRNAs during 3T3-L1 adipocyte differentiation were screened and identified by micorRNA microarray.Constructed high-expression plasmids of the adipo-related microRNAs,were transfected into the 3T3-L1 preadipocytes by lipofectamine.While the effect of adipo-related microRNAs on the course of 3T3-L1 adipocyte differentiation was observed,the protein and mRNA expression level of fatty acid binding protein(FABP4)were analyzed by Western blot and RT-PCR during 3T3-L1 adipocyte differentiation.Results The expression profiles of microRNAs have significant changed during 3T3-L1 adipocyte differentiation,in which 35 microRNAs among them down-relation,the most lowly expression is miR-24;17 microRNAs among them up-relation,the most highly expression is miR-21.MiR-24 significantly inhibited adipocyte differentiation and maturity,while miR-21 have no significant effect.MiR-24 significantly inhibited the expression of FABP4,but had no effect on the level of its mRNA;miR-21 had no effect on the expression of protein and mRNA of FABP4.Conclusion There exist adipogenic-related microRNAs during 3T3-L1 adipocyte differentiation; miR-24 play an important role in the regulation of 3T3-L1 preadipocyte differentiation into adipocyte and the(FABP4)protein expression.