Journal of immunological methods最新文献

{"title":"Inhibition of RANKL is critical for accurate assessment of anti-drug antibody incidence to denosumab in clinical studies","authors":"Mieke Jill Miller ,&nbsp;Mark A. Kroenke ,&nbsp;Troy Barger ,&nbsp;Marta Starcevic Manning ,&nbsp;Winnie Sohn ,&nbsp;Kevin Graham ,&nbsp;Daniel T. Mytych ,&nbsp;Shalini Gupta","doi":"10.1016/j.jim.2025.113864","DOIUrl":"10.1016/j.jim.2025.113864","url":null,"abstract":"<div><div>Denosumab is an approved monoclonal antibody therapeutic for the treatment of bone loss in the postmenopausal osteoporosis and oncology settings and acts by binding and neutralizing the activity of receptor activator of nuclear factor-kappa-B ligand (RANKL). Anti-drug antibodies (ADAs) to denosumab are measured <em>via</em> a standard electrochemiluminescence- (ECL) based bridging assay. In this format, the presence of soluble RANKL (sRANKL) in clinical samples can lead to false positive results. In a denosumab bioequivalence study, approximately 50 % of the serum samples showed reactivity to denosumab in the absence of a specific reagent to sequester the sRANKL. However, upon addition of osteoprotegerin (OPG) as a reagent to neutralize the sRANKL, the overall ADA incidence was lowered to &lt;1 %. To address this RANKL reactivity over the long-term use of this assay, the performance of 3 RANKL inhibitors was evaluated using healthy donor sera samples spiked with different concentrations of positive control ADA, sRANKL, or both, in an ECL based immunoassay utilizing the Meso Scale Discovery (MSD) platform. Based on these data, the denosumab antibody assay was modified to include a neutralizing anti-RANKL monoclonal antibody to eliminate the assay reactivity or false positivity due to sRANKL. Use of an anti-RANKL antibody did not impact the ADA-specific signal but inhibited the false positive assay signal due to sRANKL, resulting in an accurate detection of ADA incidence. Therefore, inhibition of interference posed by sRANKL in study samples is critical for the accurate assessment of ADA incidence towards denosumab and any biosimilars for this product that are undergoing clinical development.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"540 ","pages":"Article 113864"},"PeriodicalIF":1.6,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143839487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancing multiplex diagnostics: A novel framework for simultaneous detection of diverse pathogens using respiratory RNA viruses as a model system","authors":"M. Caputo ,&nbsp;S. Ginart ,&nbsp;L. Garrigos ,&nbsp;D. Corach ,&nbsp;F. Remes Lenicov ,&nbsp;A. Sala","doi":"10.1016/j.jim.2025.113857","DOIUrl":"10.1016/j.jim.2025.113857","url":null,"abstract":"<div><div>Numerous infectious pathologies share clinical presentation but require different management. The search for multiple pathogens in a single test could improve the resolution of these cases. For example respiratory infections, caused either by viral, bacterial or fungal agents. Rapid differential detection of common respiratory RNA viruses could be benefit by such a strategy.</div><div>We present a diagnostic method based on real-time quantitative PCR (qPCR) coupled with high-resolution melt (HRM) analysis. While applicable to diverse pathogens, this study focuses on detecting three respiratory RNA viruses: influenza A (H1N1), respiratory syncytial virus (RSV), and SARS-CoV-2.</div><div>To develop a rapid, specific, and cost-effective system, we designed a multiplex assay using SYTO 9 fluorescent dye and pathogen-specific primers. Post-amplification HRM analysis generated melt curves, with first-derivative plots enabling discrimination of the three viruses.</div><div>Samples tested in quintuplicate (10¹–10³ copies/μL) yielded distinct melt peaks. Mean melting temperatures (average Tm ± SD) were 83.64 ± 0.08 °C (SARS-CoV-2), 80.45 ± 0.07 °C (influenza H1N1), and 75.99 ± 0.07 °C (RSV). The system robustly differentiated all three viruses (100 % accuracy). Preliminary validation with clinical samples showed strong agreement with a commercial kit: κ = 0.87 (95 % CI: 0.61–1.00) for SARS-CoV-2 and κ = 1.00 (95 % CI: 0.74–1.00) for RSV.</div><div>Therefore, the designed method represents a rapid and cost-effective alternative for differential diagnosis in a single reaction tube.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"540 ","pages":"Article 113857"},"PeriodicalIF":1.6,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143795603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel combination mode for inhalant allergen screening based on 8-well microplates by LiCA","authors":"Yixian Li ,&nbsp;Xiaoming Cui ,&nbsp;Xiaohui Yang ,&nbsp;Qinqin Liu ,&nbsp;Yuanmin Sun ,&nbsp;Xue Li ,&nbsp;Huiqiang Li ,&nbsp;Yang Yu","doi":"10.1016/j.jim.2025.113856","DOIUrl":"10.1016/j.jim.2025.113856","url":null,"abstract":"<div><h3>Objective</h3><div>Currently, allergen detection is primarily classified into two types: single detection and multiple combined detection. The challenges include large volume of serum required, high testing costs, and fixed combinations of test items. A cost-effective method that uses a small amount of serum and allows for the flexible combination of allergen screening is necessary. Light-initiated Chemiluminescence Assay (LiCA) requires no washing, only need little serum, and supports flexible combination detection of allergens.</div></div><div><h3>Methods</h3><div>Based on the advantages of LiCA technology, which does not require washing, the ‘4 + 3X + 1’ allergen detection mode was established in this study. The antigens are coated on chemibeads, which are selectively added into the detection wells to achieve flexible combination, supporting the simultaneous detection of eight allergens.</div></div><div><h3>Results</h3><div>This study introduces a new allergen screening model termed “4 + 3X + 1”, where “1” represents tIgE, “4” comprises four commonly encountered indoor inhalable allergens, and “3X” encompasses a variety of frequent inhalable allergens. Allergens from “3X” could be selected based on regional characteristics and patient-specific conditions. We tested the intermediate precision(3.59 %–9.71 %) and repeatability(2.81 %–9.31 %) of the four allergens in “4-fixed”, and LoB(0.035–0.066 kU_A/L), LoD(0.092–0.156 kU_A/L), and LoQ(0.135–0.199 kU_A/L) all showed good results. We compared the consistency of LiCA and ImmunoCAP/Dot-ELISA, each of which was greater than 0.8828. In addition, two allergens were selected from “4-fixed” and “3X-optional “respectively to detect the sensitivity(80 %–97.72 %) and specificity(93.75 %–100 %).</div></div><div><h3>Conclusions</h3><div>The proposed combined test model offers a new, economical, and rapid option for allergen screening that requires low blood volumes and can be customized for individual patients.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"540 ","pages":"Article 113856"},"PeriodicalIF":1.6,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143795601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Comprehensive normalization and binary classification methods for enhanced sensitivity and reproducibility in Luminex assay quantitation” [Journal of Immunological Methods 538 (2025) 113826]","authors":"B.A. Burns ,&nbsp;C.A. Shaw ,&nbsp;M. Chandra ,&nbsp;C.S. Forconi ,&nbsp;A.M. Moormann ,&nbsp;V. Konduri ,&nbsp;V.N. Tubman ,&nbsp;W.K. Decker","doi":"10.1016/j.jim.2025.113852","DOIUrl":"10.1016/j.jim.2025.113852","url":null,"abstract":"","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113852"},"PeriodicalIF":1.6,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143692411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of polyreactive antibodies by high throughput enzyme-linked immunosorbent assay and surface Plasmon resonance","authors":"Luis Antonio Rodriguez Carnero ,&nbsp;Daniel Bedinger ,&nbsp;Simon Cocklin ,&nbsp;Jianquan Li ,&nbsp;M. Frank Erasmus ,&nbsp;Sara D'Angelo ,&nbsp;Camila Leal-Lopes ,&nbsp;Andre Azevedo Reis Teixeira ,&nbsp;Fortunato Ferrara ,&nbsp;Andrew Raymon Morton Bradbury","doi":"10.1016/j.jim.2025.113855","DOIUrl":"10.1016/j.jim.2025.113855","url":null,"abstract":"<div><div>The assessment of polyreactivity is usually carried out by enzyme linked immunosorbent assay (ELISA) using biochemically diverse target antigens with different biochemical properties, including charge and hydrophobicity, and comprising proteins, carbohydrates, nucleic acids and lipids, some of which are heterogenous in nature. Here we explored polyreactivity ELISAs based on probes of defined molecular weight, which we were also able to directly transition to a polyreactivity assay using surface plasmon resonance (SPR). Using a panel of previously characterized clinical antibodies we obtain results compatible with previous polyreactivity studies, but with potential for high throughput analysis following kinetic measurements in the early discovery process. We find ELISA is more sensitive for the detection of polyreactivity in antibodies, and with potential lower throughput, compared to SPR, but may lack the linear sensitivity of SPR.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113855"},"PeriodicalIF":1.6,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143743167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Flow cytometry reveals constant lymphocyte proportions after long-term cryopreservation of whole blood in TransFix® cell stabilization reagent","authors":"Maria Faresjö ,&nbsp;Junko Johansson ,&nbsp;Ulrika Islander ,&nbsp;Andrea Tompa","doi":"10.1016/j.jim.2025.113853","DOIUrl":"10.1016/j.jim.2025.113853","url":null,"abstract":"<div><div>Flow cytometry is an important technique for characterization of immune cells, with accurate lymphocyte profiling being essential for clinical diagnostics and research applications. While immediate processing of blood samples is ideal, long-term storage solutions are needed for large-scale studies or settings without immediate access to laboratory facilities. TransFix® is a chemical stabilization solution that allows delayed analysis by preserving cell morphology and surface markers. However, the impact of long-term cryopreservation in TransFix® on lymphocyte integrity remains underexplored. In this study, we evaluated the efficacy of cryopreservation in TransFix® for maintaining the proportions of key lymphocyte subsets, including CD3⁺ T cells, CD3⁺CD4⁺ T helper cells, CD3⁺CD8⁺ cytotoxic T cells, CD19⁺ B cells, and CD16⁺/CD56⁺ natural killer cells. Blood samples were cryopreserved in TransFix® for varying time periods, up to 48 months, and compared to fresh samples using flow cytometry. The results show that the proportions of lymphocyte subsets remain stable during cryopreservation for up to 48 months, with no significant differences observed between fresh and cryopreserved samples. This suggests that TransFix® can successfully preserve lymphocyte integrity for long-term storage, providing a reliable option for delayed analysis. These results highlight the usefulness of TransFix® for studies that require extended storage, making it easier to conduct immune monitoring in a wide range of settings.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113853"},"PeriodicalIF":1.6,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143697791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MARMOT: A multifaceted R pipeline for analysing spectral flow cytometry data from subcutaneously growing murine gastric organoids","authors":"Lydia Kirsche ,&nbsp;Jiazhuo He ,&nbsp;Anne Müller ,&nbsp;Peter Leary","doi":"10.1016/j.jim.2025.113854","DOIUrl":"10.1016/j.jim.2025.113854","url":null,"abstract":"<div><div>The analysis of murine immune cell types is a critical component of immunological research, necessitating precise and reproducible methodologies. Here, we present a comprehensive protocol and pipeline designed to streamline the process from murine gastric organoid transplant sample preparation to figure generation. This pipeline includes a detailed staining panel tailored for murine immune cells, ensuring accurate and comprehensive identification of various cell types. Additionally, it integrates an R-based analysis script (<strong>MARMOT</strong> Pipeline), encompassing data processing and visualisation. A key feature of this pipeline is its ability to produce publication-quality figures with minimal direct R coding, thus making advanced data analysis accessible to researchers with limited programming experience. Additionally, figures can be customised using a provided Shiny application. This approach both enhances the efficiency of data analysis and enables the reproducibility required for high-quality scientific research.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"540 ","pages":"Article 113854"},"PeriodicalIF":1.6,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143692416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The generation of a novel HER2-targeting nano PE25 immunotoxin with superior anti-tumor activity and high productivity","authors":"Jianguo Xiao ,&nbsp;Xianfei Chen ,&nbsp;Yibin Wan ,&nbsp;Zhenhua Guo ,&nbsp;Xiu Ren ,&nbsp;Haomin Huang","doi":"10.1016/j.jim.2025.113849","DOIUrl":"10.1016/j.jim.2025.113849","url":null,"abstract":"<div><div>Immunotoxins have the potential to be developed into anti-tumor drugs due to their targeting and strong tumor-killing activity. However, at present, issues such as dose limitations due to off-target toxicity and immunogenicity hinder the clinical use of immunotoxins. A novel HER2-targeted immunotoxin F02 was devised in this study. F02 links an anti-HER2 single-domain camelid antibody to a domain III mutant of PE toxin via a cleavable linker. The PE domain III mutant has seven point-mutations, which potentially remove B-cell and T-cell binding epitopes, thus reducing immunogenicity risks. F02 maintains anti-tumor activity in vivo and in vitro. In animals, F02 effectively inhibited the growth of NCI-N87 tumors at 1.0 mg/kg, and showed a dose-effect relationship, as the effect of completely removing tumors could be achieved at doses above 2.5 mg/kg. F02 also has low toxicity. In cynomolgus monkey models, good tolerability was observed even at 5 mg/kg, a much higher dose than the effective dose. In addition, the molecule has good druggability and can achieve soluble expression in <em>E. coli</em> with a high expression level. Thus, this new molecule has the potential to become a new option for treatment of HER2-positive tumors.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113849"},"PeriodicalIF":1.6,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NADPH oxidase subunit p22phox: A marker of oxidase-dependent oxidative stress and target for stress suppression in nonphagocytic cells","authors":"Kei Miyano ,&nbsp;Shuichiro Okamoto ,&nbsp;Fumiya Ojima ,&nbsp;Yasuhiro Takenouchi ,&nbsp;Risa Yamamoto ,&nbsp;Kimika Matsui ,&nbsp;Misaki Azuhata ,&nbsp;Mariko Inoue ,&nbsp;Mizuho Kajikawa ,&nbsp;Akira Yamauchi ,&nbsp;Futoshi Kuribayashi ,&nbsp;Shin-Ichiro Nishimatsu","doi":"10.1016/j.jim.2025.113850","DOIUrl":"10.1016/j.jim.2025.113850","url":null,"abstract":"<div><div>Reactive oxygen species (ROS)-producing NADPH oxidase (Nox) family proteins are involved in host defense. The overproduction of ROS leads to oxidative stress, which is associated with a myriad of diseases. The Nox subunit p22^{<em>phox</em>} is essential for Nox1–4 activity, and p22^{<em>phox</em>} and Nox2 proteins are mutually stabilized in phagocytic cells. This study investigated the suitability of p22^{<em>phox</em>} protein as a marker of Nox activity. To avoid contamination by phagocytic p22^{<em>phox</em>}, we developed global <em>Cybb</em> (encoding Nox2)-knockout mice and analyzed p22^{<em>phox</em>} stability and the expression profiles of Nox proteins in lysates of various tissues. We found that consistent with Nox2 in phagocytic cells, p22^{<em>phox</em>} protein was detected when Nox1–4 were coexpressed in nonphagocytic cells. Furthermore, p22^{<em>phox</em>} protein degradation was suppressed by Nox1–4, suggesting that p22^{<em>phox</em>} is a suitable marker of Nox family-dependent oxidative stress. Thus, we examined p22^{<em>phox</em>} protein levels in tissue lysates prepared from <em>Cybb</em>-knockout mice to avoid the contamination of phagocytic p22^{<em>phox</em>}. <em>Cybb</em>-knockout mice show moderately reduced p22^{<em>phox</em>} protein levels in lung tissue, suggesting that Nox2 and other Nox family members stabilized p22^{<em>phox</em>} protein. Paradoxically, this result implied that p22^{<em>phox</em>} knockdown concurrently suppressed various Nox family-dependent oxidative stress mechanisms, and this was confirmed by the suppression of Nox family-dependent directed migration in p22^{<em>phox</em>}<em>-</em>knockdown A549 human lung epithelial cells. Therefore, p22^{<em>phox</em>} not only served as a marker of Nox-dependent oxidative stress but also as a target to suppress this stress in various tissues and cells.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113850"},"PeriodicalIF":1.6,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143620864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of IgG1 immune complexes to stimulate cytokine production in innate cells","authors":"Arpita Deb,&nbsp;Kailyn Lott,&nbsp;Audrey Miceli,&nbsp;Barbara L.F. Kaplan","doi":"10.1016/j.jim.2025.113851","DOIUrl":"10.1016/j.jim.2025.113851","url":null,"abstract":"<div><div>Fcγ receptors are key immunoreceptors, that when bound by IgG immune complexes, trigger activation of downstream signaling pathways. However, there are limited in vitro assays that stimulate innate cells via Fcγ receptors that allow for evaluation of drugs or chemicals on antibody-triggered signaling. Our study investigated the activation of Fcγ receptors in innate cells using immune complexes. Our findings revealed that immobilized IgG antibodies did not elicit a significant immune response, so we designed two IgG1 immune complexes: trinitrophenyl-bovine serum albumin (TNP-BSA)/TNP-IgG1 and streptavidin-biotinylated IgG1 (Strept-Biotin IgG1). Strept-Biotin IgG1 immune complex was particularly effective, significantly enhancing IL-6, TNFα, and C3a levels, whereas TNP-BSA/TNP-IgG1 immune complex showed a modest IL-6 increase. Both TNP-BSA/TNP-IgG1 and Strept-Biotin IgG1 stimulated CD86 marker expression on F4/80+ macrophages. We also confirmed the binding of Strept-Biotin IgG1 to innate cells with fluorochrome-conjugated streptavidin. To further understand the Fcγ receptor-mediated activation of innate cells, we blocked the downstream phosphatidylinositol 3-kinase (PI3K) pathway. We found out that the PI3K inhibitor successfully suppressed IL-6 cytokine release and C3a production. However, specific Fcγ receptor-blocking antibodies failed to block IL-6 cytokine production and only modestly suppressed TNFα cytokine release, suggesting either that the antibodies were not effective blockers or that these immune complexes use other receptors. Regardless, the use of the Strept-Biotin IgG1 immune complex to stimulate cytokine production and other immune signaling was consistent with Fcγ receptor activation on innate cells which might be useful in assessing the effects of drugs or chemicals in innate cells.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113851"},"PeriodicalIF":1.6,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624607","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}