1.Mechanisms of peripheral basophils activation in patients with rheumatoid ar-thritis
Qiaofen LAN ; Huanjin LIAO ; Qiuhua CHEN ; Jun CAI ; Lifang ZHANG ; Yanxia PENG ; Yiming HE ; Ping WU ; Tong XIE ; Qingjun PAN
Chinese Journal of Immunology 2017;33(1):85-89
Objective:To investigate the activation of peripheral basophils from patients with rheumatoid arthritis ( RA) and its mechanisms. Methods:The activation markers including CD203c and the proportion of IL-4 positive rate of peripheral basophils from RA patients and healthy controls were detected by flow cytometry. Serum levels of IgE in RA patients and healthy controls were detected by electrochemilu minescence immunoassay. Basophils were negatively isolated and co-cultured with or without purified IgE,anti-IgE as positive control,then,the expression of CD203c and propotion of IL-4 positive basophils were detected by flow cytometry. Results:The expression of CD203c and IL-4 positive rate of basophils from RA patients were higher than that of healthy controls (P<0. 05). Serum levels of IgE in RA were higher than that of healthy controls (P<0. 05). After co-cultured with isolated IgE from RA patients,basophils negatively isolated from healthy controls were activated,and higher expression of CD203c and proportion of IL-4 (P<0.05). Conclusion:Basophil activation is related with development of RA,and its activation is mainly mediated by IgE. Targeting basophil and its activation pathway would be expected to provide new strategies for the treatment of RA.
2.Evaluation of the detection performance of rotavirus-enteric adenovirus fluorescent nanoparticles combined test strip
Chen LI ; Qiaofen LAN ; Huanjin LIAO ; Shangmei LI ; Ling YE ; Huiying SHENG ; Qingjun PAN
Chinese Journal of Experimental and Clinical Virology 2015;29(4):368-370
Objective To evaluate the sensitivity and specificity of rotavirus and enteric adenovirus antigen fluorescent nanoparticles rapid combined detection test strip.Methods The specimen including rotavirus,enteric adenovirus,and other enteric pathogens and 140 cases of patients with symptoms of gastrointestinal infection.The performance was analyzed after fluorescent nanoparticles rapid combined detection test strip and coloredmicrospheres rapid detection test strips were used to detect the same sample.Result The sensitivity of fluorescent nanoparticles rapid combined detection test strip is significantly higher than coloredmicrospheres rapid test strip and no specific reaction with Coxsachievirus (type A16),ECHO virus (type 30) and enterovirus (type 71).Conclusion The sensitivity of fluorescent nanoparticles rapid combined detection test strips is significantly higher than coloredmicrospheres rapid detection test strip,and the specificity is good.
3.Expression and relevance of serum IgE,IgE receptors and autoantibodies in patients with chronic urticaria
Yue YIN ; Chuang GAO ; Hong DANG ; Wen XU ; Xia PENG ; Yuting LIANG ; Huanjin LIAO ; Li LI
Chinese Journal of Laboratory Medicine 2020;43(4):411-415
Objective:To detect serum levels of immunoglobulin E (IgE), soluble version of the IgE-binding alpha-chain of Fcepsilon-RI (sFcεRIα), immunoglobulin G (IgG) anti-IgE, IgG anti-FcεRI in patients with chronic urticaria(CU), and to evaluate their relevanceMethods:Forty-three newly diagnosed patient with CU were enrolled. Thirty-seven patients with dermatitis or eczema and 51 healthy subjects were selected as the disease control (DC) and normal control (NC) group, respectively.Serum IgE was detected by immunoturbidimetry; serum anti-IgE, anti-FcεR Ⅰ and sFcεR Ⅰ α were determined byenzyme-linked immunosorbent assay (ELISA) methods. Statistical analysis was carried out by non-parametric test for comparisons of the above variables between groups, and by Spearman correlation analysis for assessment of relationships between the variables as well as between the variables and disease severity, and by receiver operating characteristic curve to analyze the diagnostic value of the variables.Results:Serum IgE, anti-IgE and anti-FcεRI in CU were significantly higher than that of NC.Their medians are 65.70 IU/ml vs 17.10 IU/ml,χ 2=28.541, P=0.001;0.61 vs 0.39,χ 2=27.408, P=0.001;0.64 vs 0.51, χ 2=29.102, P<0.001.Serumanti-FcεRI in CU was significantly lower than that in NC(0.64 vs 0.83,χ 2=25.869, P=0.007).The medians of serum sFcεRIα in CU, DC and NC groups were 5.74,5.38,4.50 ng/ml, respectively. The difference was not statistically significant,χ 2=3.463, P=0.177.There was a positive correlation between IgE and sFcεR Ⅰ α, anti-IgE and anti-FcεRI( r=0.455, P<0.001; r=0.611, P<0.001).No significant correlation was showed between the four variables and the course of disease or the severity of symptoms, P>0.05. The diagnostic performances of IgE, anti-FcεRI for CU were similar (AUC=0.72),which were better than that of sFcεRIα (AUC=0.61).The highest diagnostic efficacy (AUC=0.83) can be achieved by four joint tests.Anti-FcεRI is of value in differential diagnosis of CU and DC, AUC=0.7, P=0.002. Conclusion:The levels of serum IgE, anti-IgE, anti-FcεRI were significantly increased in CU patients, and these mast cell activation-related molecules have the potential to be diagnostic markers for CU.
4.Analysis of bone marrow morphological characteristics in acute myeloid leukemia with positive RUNX1-RUNX1T1 fusion gene
Mengna LIU ; Hongling WANG ; Huanjin LIAO ; Li LI ; Ping BAI
International Journal of Laboratory Medicine 2024;45(17):2081-2086
Objective To investigate the morphological characteristics of bone marrow in RUNX1-RUNX1T1 fusion gene-positive AML.Methods From January 2017 to January 2023,20 cases of RUNX1-RUNX1T1 fusion gene-positive AML patients newly diagnosed in this hospital were selected as group A,20 cases of AML patients without fusion gene were selected as group B,and 24 cases of PML-RARA fusion gene-positive AML patients were selected as group C.Results of bone marrow morphology,immunology,cytogenet-ics and molecular biology were analyzed retrospectively.The morphological characteristics in similar cases combined with domestic and foreign literature were summarized.Results The number of promyelocytes in group A increased in different degrees,the difference was statistically significant compared with group B(P<0.05),and the difference was also statistically significant compared with group C(P<0.001).In group A,some promyelocytes had abnormal morphology,and the mRNA level of fusion gene was positively correlated with the number of promyelocytes(r=0.478,P=0.039).Patients with expressions of CD56,CD19 and c-Kit mutation in group A was much more than that in group B(P<0.05).No significant difference was detected in remission rate after the first course of chemotherapy between group A and group B(P>0.05).Besides,the simultaneous expression of CD56 and CD19 in group A was easy to accompany the loss of sex chromosome.Conclusion AML with positive RUNX1-RUNX1T1 fusion gene could be identified with bone marrow mor-phology,immunology,cytogenetics and molecular biology.Granulocyte in the bone marrow is prone to differ-ent degrees of pathological hematopoiesis with great variability.The increased number and abnormal morphol-ogy of promyelocytes in bone marrow may be one of the morphologic features of AML with positive RUNX1-RUNX1T1 fusion gene.
5. An indirect ELISA method for quantitative detection of anti-IgE autoantibodies
Yue YIN ; Hong DANG ; Chuang GAO ; Xia PENG ; Yuting LIANG ; Huanjin LIAO ; Li LI
Chinese Journal of Laboratory Medicine 2019;42(9):782-786
Objective:
The accurate measurement of anti-IgE levels in patients′ serum helps for make diagnosis of chronic urticaria (CU). An indirect ELISA method for quantitative detection of IgG anti-IgE, were established.
Methods:
The serum samples and the clinical data of 75 first-diagnosed CU patients and 120 healthy controls were collected at Shanghai General Hospital during the year of 2018. In the indirect ELISA system to measure the IgG (anti-human IgE), the antigen, Human IgE Myeloma, was coated on the plate; Omalizumab (a humanized anti-human IgE antibody) was the standard, and horse radish peroxidase (HRP)-labeled anti-human IgG was the tracer. The optimum concentrations of serum and HRP-labeled anti-human IgG were determined by chessboard titration, and method evaluation was conducted. The comparison of serum anti-IgE levels in CU patients and healthy people were analyzed by Mann-Whitney U test and chi-square test. Receiver operating characteristic (ROC) curves were applied to establish the diagnostic performance of serum anti-IgE for CU.
Results:
The coating concentration of IgE was 5.0×10-4 mg/ml; serum dilution was 1∶300; enzyme-labeled antibody dilution was 1∶100 000. Standard curve was in good linearity with R2=0.996. The intra-and inter-assay coefficient of variation were 3.9%-7.5% and 6.0%-8.2%, and the recovery rate of low and high concentration samples were 95.9% and 108.4%, respectively. When hemoglobin≤1.3 g/L, triglyceride≤4.6 mmol/L, bilirubin≤171 μmol/L, no interference were observed. The limit of blank, limit of detection, and limit of quantitation were 5.8×10-4, 1.8×10-3, and 2.0×10-3 mg/ml. The linearity range was from 2.0×10-3 to 354.4 mg/ml. No Hook effect was found until anti-IgE reached 354.4 mg/ml. The anti-IgE remained stable after serum storage at 4 ℃ overnight or treated with 6 repeated freeze-thaw cycles. The anti-IgE levels in CU patients [19.0(1.9, 58.6)mg/ml] were significantly higher than those in healthy controls [0.7(0.002, 11.1)mg/ml] with P<0.001. When cut-off value was set as 15.3 mg/ml, in this method, the positive rate of CU patients (54.7%) was significantly higher than these of healthy controls (18.3%) (P<0.001, AUC=0.736).
Conclusions
The indirect ELISA method for serum anti-IgE measurement was successfully established. Anti-IgE autoantibodies in serum would be used in the diagnosis of CU.