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关键词： Diabetic nephropathy   IL-11   TGF-β.   mesenchymal stem cells  

摘要： OBJECTIVE:This study aims to explore the therapeutic potential of mesenchymal stem cells (MSC) in treating diabetic nephropathy (DN) by investigating their effect on IL-11 modulation in a mouse model. METHODS:The effects of MSC therapy on DN were examined both in vivo and in vitro. Sixty adult male C57BL/6 mice were divided into the streptozotocin (STZ) diabetes (T1D) and the high-fat diet diabetes (T2D) models, with both groups receiving MSC treatment or saline for 4 or 8 weeks. Blood glucose, serum urea, interleukin-11 (IL-11), and kidney fibrosis markers were measured. Additionally, western blotting was used to assess levels of Type I and III collagen, E-Cadherin, α- smooth muscle actin (α-SMA), Vimentin, and ferroptosis suppressor protein 1 (FSP-1). RESULTS:MSC-treated T1D and T2D mice showed reduced blood glucose, serum urea, IL-11, TGF-β, and fibrosis markers (type I and III collagen, α-SMA, Vimentin, FSP-1), alongside increased E-Cadherin expression. Similar effects were observed in vitro using mouse glomerular epithelial cells, confirming MSC-mediated suppression of fibrosis pathways. CONCLUSION:MSC therapy improves nephropathy, likely by inhibiting IL-11 and reducing fibrosis- related markers, making it a promising treatment for DN.

关键词： CXCL1   Equol   IL1B   Mesenchymal stem cells   NF-κB   Oxidative stress   Reactive oxygen species  

摘要： Oxidative stress (OS) inhibits the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). Equol (Eq), a phytoestrogen, exhibits notable antioxidant properties and potential for preventing osteoporosis. However, the research on the regulatory effects of Eq on stem cell osteogenesis remains limited. This investigation aimed to identify whether Eq could protect the osteogenic potential of hPDLSCs under HO-induced oxidative microenvironment. We employed a series of assays, including CCK-8, DCFH-DA, ALP staining, ARS, RT-qPCR, and Western Blotting, to assess the changes in cell viability, antioxidant capacity, and osteogenic potential following HO and Eq treatments. Our findings indicated that low concentrations of Eq had no cytotoxic effects on hPDLSCs and promoted their proliferation. Eq pre-treatment (0.5 μmol/L) partially counteracted the inhibitory effect of HO, reduced the generation of reactive oxygen species, and increased glutathione levels, thereby inhibiting oxidative damage. Eq suppressed the HO-induced inhibition of osteogenic differentiation, presenting as restoring the alkaline phosphatase levels and calcium nodule formation, as well as by upregulating the expression of BMP2 and RUNX2. Furthermore, bioinformatics analysis in this study suggested that the IL1B/NF-κB/CXCL1 signaling pathway might be a key pathway for Eq's enhancement of osteogenic differentiation potential of hPDLSCs under OS conditions. The activation of this axis by HO, which Eq can alleviate, was confirmed by validation experiments. This study provides new insights into the potential therapeutic application of Eq in alveolar bone resorption and bone regeneration research.

关键词： 人H组轮状病毒   杆状病毒表达系统   VP7蛋白   多克隆抗体  

摘要： 目的 表达和纯化人H组轮状病毒(group H rotavirus, RVH)J19株的VP7蛋白，制备VP7的多克隆抗体(多抗)并对其多抗功能进行鉴定。 方法 通过杆状病毒系统表达RVH J19株的VP7蛋白，通过亲和层析和凝胶过滤层析方法进行蛋白纯化。将纯化得到的RVH J19株的VP7蛋白免疫兔子以制备VP7多抗血清，通过酶联免疫吸附测定(enzyme-linked immunosorbent assay，ELISA)、蛋白质免疫印迹法(Western blot，WB)、免疫荧光荧光试验(Immunofluorescence，IF)等对VP7多抗血清进行鉴定。 结果 纯化得到可溶的J19 VP7蛋白；制备获得J19株VP7兔多抗；ELISA显示VP7多抗具有较好的结合效价，在稀释2 048 000倍时与人RVH VP7蛋白仍有结合；WB表明多抗能够结合J19 VP7蛋白；IF显示VP7多抗可以特异识别人H组轮状病毒J19株。 结论 本研究成功得到了人H组轮状病毒J19 VP7蛋白，制备的VP7多抗可以特异性识别人H组轮状病毒，为人H组轮状病毒的检测及病毒衣壳结构功能研究提供了基础。

关键词： Akkermansia muciniphila   Dendritic cells   Group 3 innate lymphoid cells   IL-22   Retinoic acid   STAT3   Ulcerative colitis  

摘要： BACKGROUND:The interplay between gut microbiota and immune responses is crucial in ulcerative colitis (UC). Though Akkermansia muciniphila (Akk) shows therapeutic potential, the mechanisms remain unclear. This study sought to investigate differences in therapeutic efficacy among different forms or strains of Akk and elucidate the underlying mechanisms. RESULTS:Employing a dextran sulfate sodium (DSS)-induced colitis mouse model, we assessed Akk's impact on colitis using cellular cytokine analysis, immune phenotyping, proteomics, and biochemical methods. Our results suggest that treatment with live Akk effectively reduced colitis in the DSS-induced model, whereas heat-inactivated Akk did not yield the same results. Notably, Akk exhibited protective properties by promoting the secretion of IL-22 by Group 3 innate lymphoid cells (ILC3s), as evidenced by the absence of protection in IL-22 knockout mice. Additionally, Akk augmented the population of CD103CD11b dendritic cells (DCs) and enhanced their retinoic acid (RA) synthesis through the modulation of RALDH2, a crucial enzyme in RA metabolism. The depletion of RALDH2 in DCs diminished Akk's protective properties and impaired IL-22-mediated mucosal healing. Mechanistically, Akk activated RA production in DCs by enhancing the JAK2-STAT3 signaling pathway. Additionally, various strains of Akk may exhibit differing abilities to alleviate colitis, with the novel strain Am06 derived from breast milk showing consistent efficacy similar to the reference strain. CONCLUSIONS:In summary, our findings indicate that certain strains of Akk may mitigate colitis through the promotion of RA synthesis and IL-22 secretion, underscoring the potential efficacy of Akk as a therapeutic intervention for the management of UC. Video Abstract.

关键词： NLRP12 c.1382dup   Crohn's disease   IL-1 beta   NLRP3   ERK  

摘要： Whole-genome sequencing was used to identify a dominant inherited NLRP12 c.1382dup mutation in refractory familial Crohn's disease (CD) patients. Additionally, we observed a T insertion at position 1382 in the third exon of NLRP12, leading to a frameshift mutation. Isolation of peripheral blood from mutation carriers and subsequent experiments demonstrated increased interleukin (IL)-1 beta in CD patients with the NLRP12 c.1382dup mutation. However, the mechanisms by which the NLRP12 c.1382dup mutation mediates IL-1 beta remain unclear. Our research findings reveal a close correlation between elevated p-ERK levels and increased expression of NLRP3 and IL-1 beta in the presence of the NLRP12 c.1382dup mutation. Further experiments demonstrate that inhibiting pERK with PD98059 effectively reduces the production of NLRP3 and IL-1 beta. This discovery provides new insights into the pathogenesis of CD, highlighting the significant role of the ERK/NLRP3/IL-1 beta pathway in the progression of CD. Not only does this offer novel therapeutic targets for treating CD, but it also lays the groundwork for the development of treatment strategies targeting this pathway.

关键词： Chondrocytes   Circ-PDE1C   Osteoarthritis   SGTB   miR-766-3p  

摘要： BACKGROUND:Osteoarthritis (OA) is a common degenerative joint disease. Circular RNA Phosphodiesterase 1 C (circ-PDE1C, hsa_circ_0134111) has participated in the IL-1β-induced chondrocyte damages. The objective of our study was to explore the molecular mechanism of circ-PDE1C. METHODS:Circ-PDE1C, microRNA-766-3p (miR-766-3p) or Small Glutamine Rich Tetratricopeptide Repeat Co-Chaperone Beta (SGTB) expression was determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell counting kit-8 (CCK-8) assay and flow cytometry were used to analyze proliferation and apoptosis, respectively. Western blotting assay was performed for protein detection. The inflammatory cytokines were measured by Enzyme-linked immunosorbent assay (ELISA). Oxidative stress was assessed by commercial kits. Target analysis was conducted by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. RESULTS:Circ-PDE1C was abnormally overexpressed in OA tissues and IL-1β-exposed chondrocytes. Downregulation of circ-PDE1C alleviated the IL-1β-induced cell apoptosis, inflammation, extracellular matrix degradation and oxidative stress. Circ-PDE1C could interact with miR-766-3p to serve as miRNA sponge. The function of si-circ-PDE1C was attributed to the inhibition of miR-766-3p. Additionally, miR-766-3p directly targeted the 3'UTR of SGTB. The miR-766-3p upregulation impeded the IL-1β-triggered cell damages through reducing the level of SGTB. Moreover, SGTB expression was regulated by circ-PDE1C via binding to miR-766-3p in IL-1β-induced chondrocytes. CONCLUSION:Altogether, circ-PDE1C enhanced the IL-1β-induced dysfunction in chondrocytes via upregulating SGTB by targeting miR-766-3p.

关键词： Crassostrea gigas   Cytokine production   E-type prostanoid receptor   Haemocytes   Prostaglandin E2  

摘要： Prostaglandin E2 imparts diverse physiological effects on multiple cells through its actions on four distinct E-type prostanoid (EP) receptor subtypes (EP1-EP4), among which the EP4 is one of subtypes known to mediate the immune response in mammalian monocytes and macrophages. However, the precise characteristics and functions of EP4 in mollusks remain unclear. In the present study, an EP4 homologue (designated as CgEP4) was identified from oyster Crassostrea gigas. CgEP4 contained a seven-helix transmembrane domain, shared significant homology with its vertebrate homologs, and clustered with EP4s from other Mollusca by phylogenetic analysis. When CgEP4 was transfected and expressed in HEK293 cell line, the concentration of intracellular Ca was significantly increased after treatment with its agonist PGE2, while that of cAMP was not obviously changed. The mRNA transcripts of CgEP4 were dominantly expressed in haemocytes, which was 28.37-fold (p < 0.05) higher than that in hepatopancreas. By immunofluorescence analysis, CgEP4 was found to be mainly expressed in the agranulocyte subpopulation, and CgEP4 agranulocyte accounted for 18 % of total haemocytes. After Vibrio splendidus stimulation, the mRNA expression of CgEP4 in haemocytes was not obviously changed at the first 12 h, then significantly up-regulated at 24 h, which was 2.7-fold (p < 0.01) of that in control group. After PGE2 treatment in vivo, the phosphorylation of JNK (p-JNK) in haemocytes, but not the phosphorylated ERK (p-ERK), was significantly decreased, which was 0.64-fold (p < 0.05) of that in control group. Simultaneously, the mRNA expression levels of CgTNF-1 and CgTNF-2 in haemocytes dramatically down-regulated, which were 0.06-fold (p < 0.05) and 0.73-fold (p < 0.05) of that in control group at 24 h. When the EP4 antagonist Grapiprant was added with PGE2 treatment in vivo, the p-JNK in haemocytes significantly increased, concomitant with the up-regulation of expressions of CgTNF-1 and CgTNF-2, whic

关键词： Adjuvant   Interleukin-12   Interleukin-12 receptor subunit beta-2   Paralichthys olivaceus   Th1 cells differentiation  

摘要： Interleukin-12 (IL-12) regulates the differentiation of CD4 T lymphocytes into Th1 cells by binding to its receptor, thereby promoting cellular immunity. This study characterized IL-12 and its receptor β2 (IL-12Rβ2) in flounder (Paralichthys olivaceus) and investigated their interaction, effects on T cell proliferation and differentiation, and the adjuvant effects of IL-12. The recombinant IL-12 was successfully expressed, and the IL-12Rβ2 antibody was confirmed to specifically recognize IL-12Rβ2. IL-12 bound to IL-12Rβ2 at the cellular level. IL-12 stimulation increased leukocyte proliferation and the proportion of CD4/IL-12Rβ2 cells. Moreover, blocking IL-12Rβ2 with antibody reduced Th1 markers (STAT4, T-bet, IFN-γ) and increased Th2 markers (JAK3, STAT6, GATA3). Immunization with rOmpV+IL-12 significantly upregulated CD4/IFN-γ cells on day seven, peaked the sIgM B lymphocyte response in the fourth week, and enhanced survival after Edwardsiella tarda challenge. In conclusion, IL-12 signaling effectively facilitates the differentiation of Th1 cells and negatively impacts the function of Th2 cells in flounder. This study provides new insights into the immune regulation of CD4 T cells in teleosts and lays the foundation for understanding the cellular immune mechanisms of vaccines in aquaculture.

摘要： Immune checkpoint inhibitors (ICI) represent new anticancer agents and have been used worldwide. However, ICI can potentially induce life-threatening severe cutaneous adverse reaction (SCAR), such as Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN), hindering continuous ICI therapy. We examine 6 cohorts including 25 ICI-induced SJS/TEN patients and conduct single-cell RNA sequencing (scRNA-seq) analysis, which shows overexpression of macrophage-derived CXCL10 that recruits CXCR3 cytotoxic T lymphocytes (CTL) in blister cells from ICI-SJS/TEN skin lesions. ScRNA expression profiles and ex vivo blocking studies further identify TNF signaling as a pathway responsible for macrophage-derived CXCL10 and CTL activation. Based on the trajectory analysis, ICI-activated T cells from whole blood are proposed to serve as the initial cells involved in inflammation, that lead to monocytes differentiating into macrophages and increasing their susceptibility to migrate to the lesion sites. Compared with systemic corticosteroids treatment, ICI-induced SJS/TEN patients treated with biologic TNF blockade showed a significantly rapid recovery and no recurrence of SCAR with continuous ICI therapy. Our findings identify that macrophage-eliciting CTL contribute to the pathogenesis of ICI-induced epidermal necrolysis and provide potential therapeutic targets for the management and prevention of SCAR induced by ICI therapy.

摘要： HDAC3 has been demonstrated to play a crucial role in the progression of various tumors and the differentiation and development of T cells. However, its impact on peripheral T cells in the development of murine lung cancer remains unclear. In this experiment, a subcutaneous lung tumor model was established in C57BL/6 mice, and tumor-bearing mice were treated with the specific inhibitor of HDAC3, RGFP966, at different doses to observe changes in tumor size. Additionally, a lung tumor model was established using hdac3cd4cre mice to investigate its mechanism. Mice injected with 10 mg/kg RGFP966 had the smallest tumor volume, while those injected with 30 mg/kg RGFP966 had the largest tumors. Flow cytometry analysis revealed that the expression of HDAC3 in splenic T cells was reduced in all groups of mice, while IFN-γ and IL-17 A were increased. Moreover, the expression of granzyme B and perforin in splenic CD8 T cells was increased in all groups of mice. Compared to the use of 30 mg/kg RGFP966 alone, the combination with anti-IL-17 A mAb reduced the infiltration of Neutrophils and exhausted T cells in mouse tumors, thereby impeding tumor development. These findings demonstrate that the use of RGFP966 or T cell-specific loss of hdac3 promotes the expression of IL-17 A in splenic T cells, leading to tumor resistance and providing insights for clinical treatment.