Takehiro Nakamura

MicroRNAs (miRNAs) exert critical roles in the majority of biological and pathological processes. Recent studies have associated miR‑150 with a number of different cancer types. However, little is known about miR‑150 targets in cervical cancer. In the present study, the HeLa human cervical cancer cell line was transfected with hsa‑miR‑150‑5p mimics, hsa‑miR‑150‑5p inhibitors or miRNA controls. miR‑150 was predicted to bind the 3'untranslated region (3'UTR) of the CDKN1B gene, which encodes the cyclin‑dependent kinase inhibitor 1B (p27^Kip1). The direct binding between miR‑150 and the 3'UTR of CDKN1B was confirmed using dual‑luciferase reporter assays. The effects of miR‑150 on CDKN1B mRNA expression, p27^Kip1 protein expression, cell cycle and cell proliferation were determined using reverse‑transcription quantitative PCR, western blot analysis, flow cytometry and WST‑8 assays, respectively. miR‑150 was demonstrated to directly target the 3'UTR of CDKN1B in transfected HeLa cells. The expression of CDKN1B mRNA and p27^Kip1 protein was reduced by miR‑150 mimics, and increased by miR‑150 inhibitors. Moreover, the overexpression of miR‑150 promoted cell cycle progression from the G0/G1 to the S phase and led to a significant increase in HeLa cell proliferation. The results of the present study indicated that miR‑150 promotes HeLa cell cycle progression and proliferation via the suppression of p27^Kip1 expression.

Monocytes are important cells in innate immunity. The early stage of the innate immunity is regulated by various cytokines produced by monocytes. We conducted a preliminary study to investigate TNFα expression by stimulating THP-1 cells with several bacterial species. The TNFα mRNA levels significantly varied, with the most potent stimulatory effects observed with P. gingivalis. In the present study, we focused on P. gingivalis and compared differences in cytokine expression profiles after the stimulation of THP-1 with E. coli. Bacterial antigen stimulation increased various cytokine gene expressions in THP-1. P. gingivalis had significantly more potent effects on the mRNA expressions of TNFα, IL-1β, and IL-10, but not of IL-12p40, than E. coli. This result suggests the potent ability of P. gingivalis to induce inflammation. THP-1 stimulated with LPS derived from both bacterial species showed that E. coli had significantly more potent effects on the expressions of TNFα, IL-1β, and IL-12p40 than P. gingivalis. The differences in the bacterial antigens and the LPS stimulation effects suggest involvements of different receptors, such as TLR-2 and -4, which recognize bacterial components. The present results suggest that the P. gingivalis somatic cell antigen stimulates a number of pattern recognition receptors at the same time as the synthesis of bacterial components, except LPS. The potent virulence of P. gingivalis and persistence of infection might be affected by differences in cytokine production. Pro-inflammatory responses are dependent not only on the bacterial type, but also bacterial components.

Cerebral vasospasm (CVS) is a major contributor to the high morbidity and mortality of aneurysmal subarachnoid hemorrhage (aSAH) patients. We measured histidine-rich glycoprotein (HRG), a new biomarker of aSAH, in cerebrospinal fluid (CSF) to investigate whether HRG might be an early predictor of CVS. A total of seven controls and 14 aSAH patients (8 males, 6 females aged 53.4 +/- 15.4 years) were enrolled, and serial CSF and serum samples were taken. We allocated these samples to three phases (T1-T3) and measured HRG, interleukin (IL)-6, fibrinopeptide A (FpA), and 8-hydroxy-2'-deoxyguanosine (8OHdG) in the CSF, and the HRG in serum. We also examined the release of HRG in rat blood incubated in artificial CSF. In contrast to the other biomarkers examined, the change in the CSF HRG concentration was significantly different between the non-spasm and spasm groups (p < 0.01). The rat blood/CSF model revealed a time course similar to that of the human CSF samples in the non-spasm group. HRG thus appears to have the potential to become an early predictor of CVS. In addition, the interaction of HRG with IL-6, FpA, and 8OHdG may form the pathology of CVS.

Issues concerning Black Companies have been extensively discussed in Japan recently. This paper aims to understand how discourses on Black Companies have been constructed and altered within the Japanese social context. To achieve this, we conducted discourse analysis using the case of Dentsu Inc., focusing especially on the multi-levels of discourse and power relations. We find that the process of power emergence in Black Companies and of values relating to the ethic of hard work on the part of employees relate to three levels of discourse: micro-discourse, meso-discourse and macro-discourse. This paper contributes to discourse studies by establishing that one discourse emerges under the influence of other discourses. As a practical contribution, this finding will assist those struggling with injustices in their organisation by enabling them to change their difficult situations via involvement with various discourses.