Polyporus polysaccharide (PPS), an active ingredient of traditional Chinese medicinal Polyporus umbellatus, has multiple biological functions, such as anti-cancer, immune-regulating and hepatoprotective activities. The purpose of this study was to investigate the mechanism of homogeneous polyporus polysaccharide (HPP) activated macrophages in the treatment of bladder cancer.
100 ng/mL Phorbol myristate acetate (PMA) was used to induce THP-1 human leukemic cells as a macrophage model. Then macrophages derived from THP-1 were treated with different concentrations of HPP (1, 10 and 100 μg/mL). Flow cytometry and RT-PCR were used to detected the expression of CD16, CD23, CD86, CD40 and interleukin (IL)-Iβ, iNOS mRNA. ELISA was used to test the change of IL-1β and TNF-α in macrophage after the treatment with HPP. The conditioned medium from HPP-polarized macrophages was used to detect the effect of activated macrophages on bladder cancer. MTT assay, 5-ethynyl-2'-deoxyuridine assay, flow cytometry, Transwell assay, and Western blot analysis were used to detect the effects of polarized macrophages on the viability, proliferation, apoptosis, and migration of bladder cancer cells. Western blot was also used to analysis the change of JAK2/NF-κB pathway protein.
HPP promoted the expression of pro-inflammatory factors, such as IL-Iβ, TNF-α and iNOS, and surface molecules CD86, CD16, CD23, and CD40 in macrophages and then polarized macrophages to M1 type. Results demonstrated that activated macrophages inhibited the proliferation of bladder cancer cells, regulated their apoptosis, and inhibited migration and epithelial-mesenchymal transformation (EMT). JAK2/NF-κB pathways were downregulated in the anti-bladder cancer process of activated macrophages.
The findings indicated that HPP inhibited the proliferation and progression of bladder cancer by the polarization of macrophages to M1 type, and JAK2/NF-κB pathway was downregulated in the process of anti-bladder cancer.

Follicular mature (FM) and germinal center (GC) B cells underpin humoral immunity, but the dynamics of their generation and maintenance are not clearly defined. Here, we exploited a fate-mapping system in mice that tracks B cells as they develop into peripheral subsets, together with a cell division fate reporter mouse and mathematical models. We find that FM cells are kinetically homogeneous, recirculate freely, are continually replenished from transitional populations, and self-renew rarely. In contrast, GC B cell lineages persist for weeks with rapid turnover and site-specific dynamics. Those in the spleen derive from transitional cells and are kinetically homogeneous, while those in lymph nodes derive from FM B cells and comprise both transient and persistent clones. These differences likely derive from the nature of antigen exposure at the different sites. Our integrative approach also reveals how the host environment drives cell-extrinsic, age-related changes in B cell homeostasis.
Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

Variability in gene expression across a population of homogeneous cells is known to influence various biological processes. In model organisms, natural genetic variants were found that modify expression dispersion (variability at a fixed mean) but very few studies have detected such effects in humans. Here, we analyzed single-cell expression of four proteins (CD23, CD55, CD63 and CD86) across cell lines derived from individuals of the Yoruba population. Using data from over 30 million cells, we found substantial inter-individual variation of dispersion. We demonstrate, via de novo cell line generation and subcloning experiments, that this variation exceeds the variation associated with cellular immortalization. We detected a genetic association between the expression dispersion of CD63 and the rs971 SNP. Our results show that human DNA variants can have inherently-probabilistic effects on gene expression. Such subtle genetic effects may participate to phenotypic variation and disease outcome.

The aim of the present study was to evaluate the efficacy of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) in diagnosing mediastinal and intra-abdominal lymphadenopathies. A total of 154 patients with mediastinal and intra-abdominal lymphadenopathies were included in this retrospective study between February 2010 and March 2015. Malignancy was suspected in the patients as a result of imaging findings and EUS-FNAs were performed to confirm the diagnoses. EUS and EUS-FNA data, as well as hospital medical records, were reviewed. The accuracy of EUS-FNA was 90.8% for diagnosing malignancy and 85.6% for diagnosing benign lymphadenopathy. In combination with flow cytometry (FCM), the accuracy of EUS-FNA to determine lymphoma was 94.2%. Among the malignant lymphadenopathy cases, 80 were caused by metastasis, 19 by lymphoma and 1 by myeloid leukemia. In the 53 benign cases, EUS-FNA revealed a nonspecific inflammatory condition in 27 patients, tuberculosis in 21 patients and Castleman's disease in 5 patients. The factors revealed to be associated with malignant lymphadenopathy included the sex and age of patients, as well as the location and size of the enlarged lymph node. In particular, celiac axis lymphadenopathy was associated with malignancy (23.0% of cases of malignancy, vs. 3.8% of benign lymphadenopathy). EUS-FNA results additionally suggested that the malignant lymph nodes observed in celiac axis were more likely to result from lymphoma (42.1%; 8/19 cases) than metastasis (18.8%; 15/80 cases; P=0.039). By contrast, malignant lymph nodes observed in the mediastinum were more likely to be caused by metastasis (47.5%; 38/80 cases) than lymphoma (10.5%; 2/19 cases; P=0.004). The results of the present study suggested that EUS-FNA is accurate for differentiating between malignancy and benign lymphadenopathy. Therefore, EUS-FNA in combination with FCM analysis, as a minimally invasive and highly sensitive tool, should be routinely performed for the identification of lymphoma. Additionally, examining the enlarged celiac axis lymph nodes of elderly males, who exhibit an increased risk of malignancy, may be beneficial.

The humoral immune response after acute infection with HIV-1 is delayed and ineffective. The HIV-1 envelope protein gp120 binds to and signals through integrin α4β7 on T cells. We found that gp120 also bound to and signaled through α4β7 on naive B cells, which resulted in an abortive proliferative response. In primary B cells, signaling by gp120 through α4β7 resulted in increased expression of the immunosuppressive cytokine TGF-β1 and FcRL4, an inhibitory receptor expressed on B cells. Coculture of B cells with HIV-1-infected autologous CD4(+) T cells also increased the expression of FcRL4 by B cells. Our findings indicated that in addition to mediating chronic activation of the immune system, viral proteins contributed directly to HIV-1-associated B cell dysfunction. Our studies identify a mechanism whereby the virus may subvert the early HIV-1-specific humoral immune response.