[Study on Down-regulation of Interleukin-1ß Secretion by Inhibiting ABCC1/MRP1 Transporter].

OBJECTIVE: To screen interleukin (IL)-1ß secretion-related membrane transporters by macrophage experiment in vitro and conventional knockout mice. METHODS: THP-1 cell line was differentiated to obtain human THP-1-derived macrophages, and the primary macrophages were obtained from human peripheral blood. FVB wild-type mice with the same sex and age were used as the controls of MRP1 knockout mice. The macrophages in abdominal cavity and bone marrow of mice were cultivated. The cells were treated with ABCC1/MRP1, ABCG2/BCRP, ABCB1/P-gp, OATP1B1, and MATE transporter inhibitors, then stimulated by lipopolysaccharide and adenosine triphosphate. The secretion level of IL-1ß was detected by ELISA, Western blot, and immunofluorescence. RESULTS: After inhibiting ABCC1/MRP1 transporter, the secretion of IL-1ß decreased significantly, while inhibition of the other 4 transporters had no effect. In animal experiment, the level of IL-1ß secreted by macrophages in bone marrow of MRP1 knockout mice was significantly lower than control group (P < 0.05). CONCLUSION: ABCC1/MRP1 transporter is a newly discovered IL-1ß secretion pathway, which is expected to become a new target for solving clinical problems such as cytokine release syndrome.

Effect of azithromycin combined with ambroxol in children with Mycoplasma pneumoniae pneumonia.

OBJECTIVE: This study was designed to investigate the clinical efficacy of azithromycin combined with ambroxol in children with Mycoplasma pneumoniae pneumonia (MPP). METHODS: The clinical data of 103 children with MPP treated in Fuyang District Hospital of Traditional Chinese Medicine of Hangzhou from December 2020 to August 2021 were selected and retrospectively analyzed, and these children were divided into a control group (n=51, azithromycin treatment) and a study group (n=52, azithromycin plus ambroxol treatment) according to the different treatment methods. The immunoglobulin level, pulmonary function score, treatment efficacy, serum cytokine level, disappearance time of signs and symptoms, and myocardial enzyme indices were observed and compared between the two groups. Univariate and multivariate analyses were conducted to screen the factors affecting the prognosis of MPP patients. RESULTS: After treatment, the study group showed significantly lower levels of immunoglobulin E, immunoglobulin G, immunoglobulin M and immunoglobulin A; higher pulmonary function scores, and lower levels of interleukin-6, human interferon-gamma, and monocyte chemoattractant protein-4 compared with the control group (all P < 0.05). The total incidence of adverse reactions such as nausea, diarrhea, abdominal pain, and vomiting was 15.38% in the study group, which was slightly lower than that in the control group (17.65%), exhibiting no significant difference (P > 0.05). The disappearance time of cough and lung rales, time required to restore to a normal body temperature, and hospital stay in the study group were shorter than those in the control group (P < 0.05). After treatment, aspartate aminotransferase, lactate dehydrogenase, creatine kinase, and creatine kinase isoenzyme in the study group were lower than those in the control group (all P < 0.05). The course of disease before admission, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), anemia, albumin < 30 g/L, drug initiation time, pulmonary consolidation, and complications involving multiple systems were the main factors affecting the efficacy of azithromycin combined with ambroxol in the treatment of MPP. CONCLUSION: Azithromycin combined with ambroxol can effectively improve the immunoglobulin level and lung function, reduce the level of inflammatory factors, improve the therapeutic effect, shorten the recovery process, and reduce the degree of myocardial damage, which is effective in the treatment of MPP and is worth promoting.

A Novel NUP98/RARG Gene Fusion in Pediatric Acute Myeloid Leukemia Resembling Acute Promyelocytic Leukemia.

Here, we introduced the first case of acute myeloid leukemia (AML) with RARG-NUP98 in a pediatric patient. The young male presented with structural and functional abnormalities similar to hypergranular acute promyelocytic leukemia, but was resistant to all transretinoic acids and arsenic trioxide. Till date, only 12 adult AML cases involving RARG rearrangement have been reported. At present, there is no standardized or optimal treatment option for this AML subtype. Disease management may typically require a joint treatment strategy involving chemotherapy, immunotherapy, and support therapy. In this study, we report the clinical manifestations and experimental results of a 10-year-old male and review other cases of RARG gene rearrangement reported in the literature.

Treatment of Relapsed and Refractory ALK-Positive Anaplastic Large Cell Lymphoma With ALK-Specific Tyrosine Kinase Inhibitor in Children: A Case Series.

Relapsed and refractory ALK-positive anaplastic large cell lymphoma (ALCL) has a poor prognosis. In this report, we present 3 relapsed/refractory pediatric ALCL patients, 1 of these with central nervous system involvement. All 3 patients were treated with ALK inhibitor and achieved complete response. Both crizotinib and alectinib have shown significant activity in pediatric patients with refractory ALK-positive ALCL.

Deep Multimodal Fusion Autoencoder for Saliency Prediction of RGB-D Images.

In recent years, the prediction of salient regions in RGB-D images has become a focus of research. Compared to its RGB counterpart, the saliency prediction of RGB-D images is more challenging. In this study, we propose a novel deep multimodal fusion autoencoder for the saliency prediction of RGB-D images. The core trainable autoencoder of the RGB-D saliency prediction model employs two raw modalities (RGB and depth/disparity information) as inputs and their corresponding eye-fixation attributes as labels. The autoencoder comprises four main networks: color channel network, disparity channel network, feature concatenated network, and feature learning network. The autoencoder can mine the complex relationship and make the utmost of the complementary characteristics between both color and disparity cues. Finally, the saliency map is predicted via a feature combination subnetwork, which combines the deep features extracted from a prior learning and convolutional feature learning subnetworks. We compare the proposed autoencoder with other saliency prediction models on two publicly available benchmark datasets. The results demonstrate that the proposed autoencoder outperforms these models by a significant margin.

Efficacy and safety of immunosuppressive therapy with or without eltrombopag in pediatric patients with acquired aplastic anemia: A Chinese retrospective study.

To determine the efficacy and safety of eltrombopag (E-PAG) combined with intensive immunosuppressive therapy (IST) for the treatment of pediatric patients with severe aplastic anemia (SAA). A total of 57 pediatric patients with newly diagnosed severe aplastic anemia were enrolled in this study. Thirty nine patients were treated with IST alone, consisting of porcine anti-human thymocyte globulin (30 mg/kg/day × 5 days) and cyclosporine A (CsA) (treated for 2 years, with a trough concentration maintained at 200-250 ng/mL), and 18 patients were treated with IST + E-PAG (12.5-50 mg/day, maintained for 6 months). We found no statistical difference between the response rates at 3 months for the two groups (CR: 12.8% vs. 22.2% p > 0.05, ORR: 56.4% vs. 77.7% p > 0.05). However, we found a statistical difference between the response rates at 6 months for the two groups (CR: 17.9% vs. 50% p < 0.05, ORR: 69.2% vs. 94.4% p < 0.05). The main side-effect during treatment with E-PAG was having a slightly to moderately elevated bilirubin level, which was temporary and controllable, accounting for approximately 66.6% (12/18) of patients in the IST + E-PAG group vs. 20.5% (8/39) of those in the IST group (p < 0.05). IST + E-PAG therapy appears to be more effective than IST alone for the treatment of pediatric SAA, with good tolerability and compliance. This approach deserves further exploration.

Chimeric antigen receptor T cell therapy can be administered safely under the real-time monitoring of Th1/Th2 cytokine pattern using the cytometric bead array technology for relapsed and refractory acute lymphoblastic leukemia in children.

CD19 chimeric antigen receptor T (CD19CAR-T) cell therapy has shown striking response in treating relapsed and refractory B-lineage acute lymphoblastic leukemia (r/r B-ALL). However, side-effects including cytokine release syndrome (CRS) and neurotoxicity can be fatal to patients. In this report, five patients with r/r B-ALL were treated with CD19CAR-T cells. Cytokine release syndrome experienced by four patients who achieved complete remission (CR) with minimal residual disease (MRD) negative. One patient who did not response to the treatment had no CRS. Acute toxicities including fever, hypotension and other neurological toxicities occurred in responding patients within 2 weeks post infusion and managed properly with tocilizumab and/or steroids according to the "real-time" monitoring of a simple 6 Th1/Th2 cytokine pattern. In conclusion, our study demonstrates that CD19CAR-T cell therapy can be safely administered for patients with relapsed and refractory leukemia under the "real-time" monitoring of a simple 6-cytokine pattern.

Inhibitory effects of human immunodeficiency virus gp120 and Tat on CpG-A-induced inflammatory cytokines in plasmacytoid dendritic cells.

Plasmacytoid dendritic cells (pDCs), not only inhibit viral replication, but also play an essential role in linking the innate and adaptive immune system. In this study, we explored the effects of human immunodeficiency virus (HIV) gp120 and tat on CpG-A-induced inflammatory cytokines in pDCs. The results provided fundamental insights into HIV pathogenesis that may hold promise for preventative and even curative strategies. pDCs were isolated using blood DC antigen 4 (BDCA-4) DC isolation kit, and the purity was analyzed using BDCA-2 antibody by flow cytometry. pDCs and peripheral blood mononuclear cells (PBMCs) were stimulated by either CpG-A (5 µg/ml), gp120 (0.5 µg/ml), tat (0.5 µg/ml), or CpG-A treatment combined with gp120 or tat. The production of type I interferons (IFNs) and other inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interlukine-6 (IL-6), and interferon-gamma-inducible protein-10 (IP-10) in the culture supernatant, was determined by enzyme-linked immunosorbent assay. The results showed that CpG-A induced high levels of type I IFNs and other inflammatory cytokines, including TNF-α, IL-6, and IP-10, in pDCs. Concomitant treatment with gp120 reduced the levels of IFN-α, IFN-ß, TNF-α, IL-6, and IP-10 induced by CpG-A in pDCs by 79%, 53%, 60%, 50%, and 34%, respectively, while tat suppressed them by 88%, 66%, 71%, 64%, and 53%, respectively. Similar results were demonstrated in CpG-A-treated PBMCs. In conclusion, gp120 and tat are effective inhibitors of the CpG-A-mediated induction of type I IFNs and other inflammatory cytokines from pDCs and PBMCs.

Multiple signaling pathways are involved in the interleukine-4 regulated expression of DC-SIGN in THP-1 cell line.

Dendritic cell-specific intercellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN) is an important pattern recognition receptor on dendritic cells (DCs), and its expression shows significant cytological and histological specificity, being interleukine-4 (IL-4) dependent. The signaling pathways through which IL-4 regulates expression of DC-SIGN are still unclear. We used phorbol 12-myristate 13-acetate- (PMA-) differentiated THP-1 cells as the in vitro model of monocyte/macrophage cells to study the signaling pathways involved in IL-4-regulated expression of DC-SIGN. We found that a high expression of DC-SIGN could be induced by IL-4 at the levels of mRNA and cell surface protein. Upregulated expression of DC-SIGN was almost completely blocked by the specific inhibitor of ERK pathway, and partly reduced by the specific inhibitors of JAK-STAT and NF-κB pathways. The activation of the three signaling pathways was directly confirmed by testing the phosphorylation of protein kinase within the cytoplasm and nucleus over time. The analysis of cis-acting elements of DC-SIGN promoter showed that the activity of DC-SIGN promoter without Ets-1 transcription factors binding site almost completely disappeared. Our results demonstrated that multiple signaling pathways are involved in IL-4 induced high expression of DC-SIGN on THP-1 cells, in which ERK pathway is the main signaling pathway and mediated by the Ets-1 transcription factors binding site.

Detection of hepatitis C virus by an improved loop-mediated isothermal amplification assay.

An improved, sensitive, specific, and rapid one-step reverse transcription loop-mediated isothermal amplification (LAMP) assay targeting the 5' untranslated region (UTR) was developed to detect hepatitis C virus (HCV) infection. Based on an accelerating primer (AP), the present assay, named AP-LAMP, has the advantages of rapidity and sensitivity over the routine LAMP method. The possible AP-based amplification pathway during the reaction was revealed by restriction enzyme digestion and eletrophoresis. The detection limit of the AP-LAMP assay was approximately 84 IU/ml, and no cross-detection was observed. The assay was evaluated further with 126 clinical specimens, and the results indicated the suitability and simplicity of the test as a rapid diagnostic tool for detection of HCV RNA.

Honokiol: an effective inhibitor of tumor necrosis factor-α-induced up-regulation of inflammatory cytokine and chemokine production in human synovial fibroblasts.

In this study, we investigated the mechanisms underlying the anti-inflammatory effects of honokiol in tumor necrosis factor (TNF)-α-stimulated rheumatoid arthritis synovial fibroblasts (RASFs). RASFs pre-treated with honokiol (0-20 µM) were stimulated with TNF-α (20 ng/ml). The levels of prostaglandin E2 (PGE2), nitric oxide (NO), soluble intercellular adhesion molecule-1 (sICAM-1), transforming growth factor-ß1 (TGF-ß1), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) in supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and Griess assay. In addition, protein expression levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and phosphorylated Akt, nuclear factor kappa B (NFκB), and extracellular signal-regulated kinase (ERK)1/2 were determined by western blot. The expression of NFκB-p65 was assessed by immunocytochemical analysis. TNF-α treatment significantly up-regulated the levels of PGE2, NO, sICAM-1, TGF-ß1, MCP-1, and MIP-1α in the supernatants of RASFs, increased the protein expression of COX-2, iNOS, and induced phosphorylation of Akt, IκB-α, NFκB, and ERK1/2 in RASFs. TNF-α-induced expression of these molecules was inhibited in a dose-dependent manner by pre-treatment with honokiol. The inhibitory effect of honokiol on NFκB-p65 activity was also confirmed by immunocytochemical analysis. In conclusion, honokiol is a potential inhibitor of TNF-α-induced expression of inflammatory factors in RASFs, which holds promise as a potential anti-inflammatory drug.

[Expression of CD38 and HLA-DR on CD8+ T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART)].

OBJECTIVE: To study the expression of CD38 and HLA-DR on CD8(+) T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART) and the relationship of immune activation and disease progression. METHODS: A cross-section study of 194 pediatric AIDS patients receiving HAART was carried out and 52 age-matched healthy children were recruited as control. The percentage of CD4(+), CD8(+), CD8(+)/CD38(+) and CD8(+)/HLA-DR(+) T cells was tested using flow cytometry, and HIV-RNA in plasma was detected by quantitative RT-PCR. RESULTS: One hundred and ninety-four pediatric AIDS patients were divided into two groups according to the viral load: 59 patients with VL ≥ 400 copies/ml and 135 patients with VL < 400 copies/ml. The percentage of CD8(+)/CD38(+) and CD8(+)/HLA-DR(+) T cells of patients with VL ≥ 400 copies/ml was significantly higher than that of patients with VL < 400 copies/ml (P < 0.05). Of patients with VL < 400 copies/ml, the percentage of CD8(+)/CD38(+) T cells was nearly normal, and the percentage of CD8(+)/HLA-DR(+) T cells was higher than normal level (P < 0.05). There was a positive correlation between percentage of CD8(+)/CD38(+) and of CD8(+)/HLA-DR(+)T cells and viral load (R = 0.403, P = 0.03 for the former and R = 0.569, P = 0.09 for the later). CONCLUSIONS: Effective HAART could decrease immune activation of HIV-infected children significantly. And there was a positive correlation between percentage of CD8(+)/CD38(+) and of CD8(+)/HLA-DR(+)T cells and viral load, suggesting that the two indicators might be used as the substitution of viral load in resource-limited areas.