ObjectiveTo investigate the effects of Yishen Juanbi pills （YSJB）-containing bone marrow fluid on the migration and differentiation phenotypes of CD4⁺T lymphocytes based on the stromal cell-derived factor-1/chemokine receptor 4 （SDF-1/CXCR4） signaling pathway. MethodsPrimary CD4⁺T lymphocytes were isolated from mice using magnetic bead separation and identified for purity by flow cytometry. A CD4⁺T lymphocyte culture system was then established to observe the effects of SDF-1 on CD4⁺T-cell migration and differentiation. On this basis， the experimental groups included the Sham group， the ovariectomy （OVX） group， the Sham+collagen-induced arthritis （CIA） group， the OVX+CIA group， the Sham+CIA+YSJB group （2.16 g·kg^-1）， the OVX+CIA+YSJB group （2.16 g·kg^-1）， and the OVX+CIA+methotrexate （MTX） group （1.5 mg·kg^-1）. Bone marrow fluid from each group was prepared according to previous methods and added to the CD4⁺ T-cell culture system at 5% （v/v）. Transwell assays were used to examine CD4⁺T-cell migration in each group. Real-time PCR was used to measure the mRNA expression levels of interleukin （IL）-17， tumor necrosis factor-α （TNF-α）， retinoic-acid-related orphan receptor γt （RORγt）， IL-10， transforming growth factor-β （TGF-β）， forkhead box P3 （FoxP3）， CXCR4， phosphoinositide 3-kinase （PI3K）， and protein kinase B （Akt）. Western blot was used to detect the expression of helper T （Th）17/regulatory T （Treg） cell signature factors （RORγt， FoxP3）， CXCR4， PI3K， phosphorylated （p）-PI3K， Akt， and p-Akt. In a separate set of experiments， cells were divided into the Sham group， OVX+CIA group， OVX+CIA+CXCR4 antagonist AMD3100 group， and OVX+CIA+YSJB+AMD3100 group to observe changes in the above indicators following AMD3100 intervention. ResultsCompared with the Sham group， the number of migrated cells in the lower chamber was significantly increased in the Sham+CIA and OVX+CIA groups （P<0.05， P<0.01）. The mRNA expression of RORγt， IL-17， TNF-α， CXCR4， PI3K， and Akt was significantly upregulated， whereas FoxP3， IL-10， and TGF-β mRNA expression was significantly decreased （P<0.05， P<0.01）. Protein expression of RORγt， CXCR4， p-PI3K/PI3K， and p-Akt/Akt was significantly increased， while FoxP3 protein expression was markedly decreased （P<0.05， P<0.01）. Compared with the OVX+CIA group， the OVX+CIA+YSJB group and OVX+CIA+MTX group showed significantly reduced migration （P<0.05）， mRNA expression of RORγt， IL-17， TNF-α， CXCR4， PI3K， and Akt was also significantly decreased， while FoxP3， IL-10， and TGF-β mRNA expression was significantly increased （P<0.05， P<0.01）. RORγt protein expression was significantly downregulated， and FoxP3 protein expression markedly upregulated （P<0.05）. In the OVX+CIA+YSJB group， CXCR4， p-PI3K/PI3K， and p-Akt/Akt protein expression was significantly decreased （P<0.05）. Compared with the OVX+CIA group， RORγt， CXCR4， PI3K， and Akt mRNA expression in CD4⁺T cells was significantly decreased in the OVX+CIA+AMD3100 group and the OVX+CIA+YSJB+AMD3100 group， while FoxP3 mRNA and protein expression was significantly upregulated （P<0.05， P<0.01）. RORγt， CXCR4， p-PI3K/PI3K， and p-Akt/Akt protein expression was also markedly decreased （P<0.05， P<0.01）. Compared with the OVX+CIA+AMD3100 group， the OVX+CIA+YSJB+AMD3100 group showed significantly decreased RORγt and Akt mRNA expression （P<0.05） and significantly lower p-Akt/Akt protein expression （P<0.05）. ConclusionYSJB-containing bone marrow fluid suppresses CD4⁺T-cell migration and regulates Th17/Treg balance by downregulating Th17-associated signature factors and upregulating Treg-associated signature factors through inhibition of the SDF-1/CXCR4 signaling pathway and PI3K/Akt signaling pathway. The SDF-1/CXCR4 signaling pathway is one of the targets through which YSJB inhibits CD4⁺T-cell differentiation.

Objective: To analyze the association between polycyclic aromatic hydrocarbons(PAHs)exposure and rheumatoid arthritis(RA)based on large sample data. Methods: The RA patients(RA group)and non-RA patients(non-RA group)with complete data were selected from the National Health and Nutrition Survey Database in the United States(NHANES)(2005—2014). The logistic regression model was used to analyze the association between 8 monohydroxylated(OH-)PAH metabolites in the urine and RA. Results: A total of 357 RA patients and 5, 256 non-RA participants were included.After adjusting the confounding factors by logistic analysis, the level of OH-PAHs mixture at the highest quartile(Q4)was associated with increased risk of RA compared with that at the lowest quartile(Q1)(OR=1.60, 95%CI: 1.16-2.23). For a single kind of OH-PAHs, the Q4 levels of 1-hydroxynaphthalene(OR=1.59, 95%CI: 1.14-2.23), 2-hydroxynaphthalene(OR=1.66, 95%CI: 1.19-2.32), 2-hydroxyfluorene(OR=1.61, 95%CI: 1.17-2.22), 3-hydroxyfluorene(OR=1.64, 95%CI: 1.18-2.27)and 1-hydroxyphenanthrene(OR=1.38, 95%CI: 1.00-1.94)were all associated with significantly increased risk of RA compared with the Q1 level(all P<0.05). However, the Q2 level of 1-hydroxypyrene(OR=0.60, 95%CI: 0.43-0.83)was related to a decreased incidence of RA(P<0.01). Conclusions OH-PAHs mixed exposure is a risk factor for RA.The association between the level of individual OH-PAH and the rate of RA is bidirectional and is depended on the type and concentration of OH-PAHs.

Background: Aberrant Bcl-2 transcription is closely related with nodal diffuse large B-cell lymphoma (DLBCL), however, the relationship between Bcl-2 and primary gastrointestinal DLBCL (PGI-DLBCL) was not fully studied.Aims: To investigate the relationship between Bcl-2 gene amplification and protein expression and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL.Methods: Clinical data was collected from 136 PGI-DLBCL patients receiving surgical treatment, and a telephone interview was conducted for survival information.Bcl-2 gene amplification and protein expression in tumor tissue were determined by fluorescence in situ hybridization and immuno-histochemistry, respectively, and relationships between Bcl-2 and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL were analyzed.Results: Among 136 PGI-DLBCL patients, 33 (24.3%) showing gene amplification and 90 (66.2%) showing protein expression of Bcl-2;gene amplification was correlated with primary tumor location, Ann Arbor stage, serum lactate dehydrogenase level, B symptom and International Prognostic Index (IPI) score (P<0.05), while protein expression was correlated with primary tumor location and immunophenotype (P<0.05).5-year overall survival (OS) in patients positive for Bcl-2 gene amplification and patients with non-GCB immunophenotype and positive for Bcl-2 protein expression were inferior to those negative ones (41.5%vs.71.5%, P<0.05;54.6% vs.84.6%, P<0.05).In Bcl-2 gene amplification or protein expression positive patients, 5-year OS of CHOP chemotherapy was inferior to that of rituximab combined with CHOP chemotherapy (48.6%vs.80.3%, P<0.05;66.4%vs.83.4%, P<0.05).Conclusions: Detection of Bcl-2 gene amplification is useful for prediction of prognosis in PGI-DLBCL.Both patients with Bcl-2 gene amplification and non-GCB patients with Bcl-2 protein expression have a poorer prognosis.Rituximab may improve the prognosis in patients with Bcl-2 gene amplification or protein expression.

This study was aimed to observe the effect of Bai-Zhu Huang-Qi (BZHQ) decoction ethyl acetate extract on NOD like receptor family,pyrin domain-containing 3 (NLRP3) inflammasome in macrophages.The U937 cells were pretreated with phorbol 12-myristate 13-acetate (PMA,10 ng· mL-1) for 48 hours to induce macrophages.Effects on cell viability by different doses of BZHQ decoction ethyl acetate extract (0,3.125,6.25,12.5,25,50,100 μg· mL-1) were observed to select the appropriate concentration.Contents of NLRP3 and caspase-1 in cells were detected by real-time PCR and western blot.The concentration of interleukin-1β (IL-1β) in cell supernatant was detected by enzyme linked immunosorbent assay (ELISA).The cell counting kit-8 (CCK-8) assay showed that when the drug concentration was lower than 25 μg· mL-1,there was no impact on cell viability;when the drug concentration was higher than 50 μg· mL-1,there was inhibition on cell viability (P ＜ 0.05).The concentration of 25 μg· mL-1 was used to conduct the following experiment.Compared to the blank group,the expression of NLRP3 and caspase-l in cells of the LPS group were significantly increased (P ＜ 0.01).The concentration of IL-1β in cell supernatant was also significantly increased (P ＜ 0.01).After treated with BZHQ decoction ethyl acetate extract,levels of NLRP3,caspase-1 and IL-1β were significantly decreased (P ＜ 0.05).It was concluded that BZHQ decoction ethyl acetate extract can inhibit the production of NLRP3 inflammasome in LPS-stimulated macrophages.