便潜血陰性の鉄欠乏性貧血の1例 : 民間療法による経皮的脱血療法による失血例

患者は鉄欠乏性貧血が次第に生じてきた78歳男性である.便潜血反応は常に陰性であり,上部および下部消化管検査でも全く異常は認められず,入院精査となった.全身的な検索を終えたところ,患者は1年半前より,鍼灸師に通院しており,1ヵ月におよそ100 mlほど,経皮的に真空吸引器による混血療法を行っていたことを明らかにした.脱血治療時に上腕部,肩や背部に生じた皮下出血斑は,1ヵ月後に当院に来院するときまでには完全に消失していたため,貧血の成因が不詳であった.直ちに真空吸引療法を中止し,経口鉄剤を補給することにより,貧血は急速に改善された.患者は,最近5年間,元気に過ごしており,血清鉄もフェリチン濃度も正常範囲である.この症例は,便潜血が常に陰性の鉄欠乏性貧血患者を診た場合には現病歴を詳細に聴取することが最も重要であることを示している.A 78-year-old man insidiously developed iron-deficiency anemia. Occult blood tests of the stools remained consistently negative. Results of gastroscopic and colonoscopic examinations were normal. After systemic examinations had been performed, the patient admitted that he had been treated by an acupuncturist for the previous 18 months who had aspirated nearly 100 ml blood per month transcutaneously using a vacuum aspirator. Subcutaneous hemorrhages, which developed at the sites of vacuum-aspiration on the brachium, shoulders and back, had disappeared almost completely by the time the patient visited one month later. After the vacuum aspiration therapy had been stopped and oral administration of iron supplement started, the anemia improved rapidly. The patient has been doing well for the last 5 years, with normal serum levels of iron and ferritin. The present data indicate that history taking is very important for the differential diagnosis of iron-deficiency anemia, particularly when there is no evidence of blood loss in the gastrointestinal tract

C/EBPβ directly bound to <i>Ihh</i> promoter.

<p>(A) EMSA for specific binding of C/EBPβ to the <i>Ihh</i> promoter. Consensus oligonucleotide (C), wild-type (WT) and mutant (MT) probes were incubated with nuclear extract from C/EBPβ-transfected ATDC5 cells. Competition and supershift experiments were also performed. Data are representative of two independent experiments performed in duplicate. (B) A ChIP assay for C/EBPβ using ATDC5 cells cultured for 3 weeks. Semi-quantitative RT-PCR was performed using primers as follows: promoter region of <i>Ihh</i> (from −259 to −160) and negative control (from −1274 and −1102 bp). Data are representative of two independent experiments performed in duplicate.</p

C/EBPβ stimulated the expression of Ihh in <i>ex vivo</i> organ cultures.

<p><i>Ex vivo</i> organ culture of tibias dissected from E14.5 mouse embryos. Tibias were cultured for 4 days after transfection with adenovirus vectors expressing LacZ control (top row) and C/EBPβ-LAP (bottom row). Safranin O staining and immunofluorescent staining were performed to localize C/EBPβ, RUNX2 and Ihh. DAPI was used as a counterstain. Red, green and blue bars indicate the proliferative, pre-hypertrophic and hypertrophic zones, respectively. Scale bar, 500 µm. Histological analysis was repeated at least twice for each sample from six pairs of limbs, respectively.</p

C/EBPβ binding element is crucial for RUNX2 to regulate transcriptional activity of <i>Ihh</i>.

<p>(A) Deletion constructs were co-transfected with 0.2 µg of RUNX2 or GFP into HeLa cells. Means ± S.D. of duplicates from three independent experiments are shown. ^*<i>p</i><0.05. (B) Mutation constructs of C/EBPβ and RUNX2 binding elements in pDel3 were co-transfected with 0.2 µg of RUNX2 or GFP into HeLa cells. Means ± S.D. of duplicates from three independent experiments are shown. ^*<i>p</i><0.05. (C) EMSA for specific binding of RUNX2 to the C/EBPβ binding site of <i>Ihh</i> promoter. Wild-type (WT) probe, which harbors C/EBPβ binding site, was incubated with nuclear extract from C/EBPβ-transfected ATDC5 cells. Supershift experiment using RUNX2 antibody was also performed. Data are representative of two independent experiments performed in duplicate. (D) A ChIP assay for RUNX2 using ATDC5 cells cultured for 3 weeks. Semi-quantitative RT-PCR was performed using same primers as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104547#pone-0104547-g005" target="_blank">Figure 5B</a>. Data are representative of two independent experiments performed in duplicate. (E) Immunoprecipitation (IP) and Immunoblotting were performed. Nuclear extract was obtained from C/EBPβ-transfected ATDC5 cells. Immunoprecipitated proteins with C/EBPβ, RUNX2 or IgG antibody were subjected to SDS-PAGE and immunoblotting using C/EBPβ or RUNX2 antibody.</p