Low-grade Myofibroblastic Sarcoma at the Base of the Tongue

Low-grade myofibroblastic sarcoma (LGMS) represents a distinct atypical myofibroblastic tumor that occurs at several sites, primarily within the head and neck regions. A painless, enlarged mass is the most common clinical presentation, but a definitive diagnosis requires both histopathological and immunohistochemical analyses. Histologically, LGMS commonly presents as a cellular lesion composed of spindle-shaped tumor cells arranged primarily in fascicles with a diffusely infiltrative pattern. Immunohistochemically, LGMS shows positive staining for at least one myogenic marker, such as desmin or muscle actin. Here we report a case of LGMS in the base of the tongue. Our case showed positive immunostaining for desmin and vimentin, and was thus diagnosed as LGMS. The patient received surgery, but no chemotherapy or radiotherapy, and was completely without evidence of the disease 38 months after the surgery

Recombinant human FGF-2 for the treatment of early-stage osteonecrosis of the femoral head: TRION, a single-arm, multicenter, Phase II trial

Aim: This study aimed to evaluate the 2-year outcomes from a clinical trial of recombinant human FGF-2 (rhFGF-2) for osteonecrosis of the femoral head (ONFH). Patients & methods: Sixty-four patients with nontraumatic, precollapse and large ONFHs were percutaneously administered with 800 μg rhFGF-2 contained in gelatin hydrogel. Setting the end point of radiological collapse, we analyzed the joint preservation period of the historical control. Changes in two validated clinical scores, bone regeneration and safety were evaluated. Results: Radiological joint preservation time was significantly higher in the rhFGF-2 group than in the control group. The ONFHs tended to improve to smaller ONFHs. The postoperative clinical scores significantly improved. Thirteen serious adverse events showed recovery. Conclusion: rhFGF-2 treatment increases joint preservation time with clinical efficacy, radiological bone regeneration and safety

Involvement of crosstalk between Oct4 and Meis1a in neural cell fate decision.

Oct4 plays a critical role both in maintaining pluripotency and the cell fate decision of embryonic stem (ES) cells. Nonetheless, in the determination of the neuroectoderm (NE) from ES cells, the detailed regulation mechanism of the Oct4 gene expression is poorly understood. Here, we report that crosstalk between Oct4 and Meis1a, a Pbx-related homeobox protein, is required for neural differentiation of mouse P19 embryonic carcinoma (EC) cells induced by retinoic acid (RA). During neural differentiation, Oct4 expression was transiently enhanced during 6-12 h of RA addition and subsequently disappeared within 48 h. Coinciding with up-regulation of Oct4 expression, the induction of Meis1a expression was initiated and reached a plateau at 48 h, suggesting that transiently induced Oct4 activates Meis1a expression and the up-regulated Meis1a then suppresses Oct4 expression. Chromatin immunoprecipitation (ChIP) and luciferase reporter analysis showed that Oct4 enhanced Meis1a expression via direct binding to the Meis1 promoter accompanying histone H3 acetylation and appearance of 5-hydoxymethylcytosine (5hmC), while Meis1a suppressed Oct4 expression via direct association with the Oct4 promoter together with histone deacetylase 1 (HDAC1). Furthermore, ectopic Meis1a expression promoted neural differentiation via formation of large neurospheres that expressed Nestin, GLAST, BLBP and Sox1 as neural stem cell (NSC)/neural progenitor markers, whereas its down-regulation generated small neurospheres and repressed neural differentiation. Thus, these results imply that crosstalk between Oct4 and Meis1a on mutual gene expressions is essential for the determination of NE from EC cells

Stimulatory effect of Meis1a on the formation of neurospheres consisted of NSCs/neural progenitor cells.

<p>(<b>A</b>) Effect of Meis1a on sphere formation during neural differentiation. Aggregated S-Meis1a and AS-Meis1a were treated with RA in the presence or absence of MIF. After 4 days, spheres were analyzed under a phase-contrast microscope. <i>Scale bar</i> = 100 µm. (<b>B</b>) Quantification of sphere sizes of S-Meis1a and AS-Meis1a indicated in <b>A</b>. More than 400 spheres of each sample were analyzed. (<b>C</b>) Effect of Meis1a on the cell growth during neural differentiation. (<b>D</b> and <b>E</b>) Aggregated S-Meis1a and AS-Meis1a cells were treated with RA together with or without MIF. Effects of Meis1a on the NSC/neural progenitor marker expressions. Cell lysates from RA-primed S-Meis1a (<b>D</b>) and AS-Meis1a (<b>E</b>) cells with or without MIF were analyzed by WB with anti-Nestin, anti-GLAST, anti-BLBP and anti-Sox1 antibodies. (<b>F</b> and <b>G</b>) Quantification of expression levels of NSC/neural progenitor markers in S-Meis1a and AS-Meis1a cells as shown in <b>D</b> and <b>E</b>, respectively.</p

Effect of Meis1a on Sox2 and Pax6 expressions.

<p>(<b>A</b>) Expression patterns of Sox2 and Pax6 during neural differentiation. Aggregated P19 cells were treated with RA for various times and analyzed by WB with anti-Sox2 and Meis1 antibodies. (<b>B</b>) Quantification of expression levels of Sox2 and Pax6 as shown in <b>A</b>. (<b>C</b> and <b>D</b>) Stimulation of <i>Sox2</i> and <i>Pax6</i> mRNA and protein expressions by ectopic expression of Meis1a. Monolayer-cultured P19 cells were transfected with various amounts of pcDNA3-EF1-α-<i>Meis1a</i> and after 12 h, <i>Sox2</i> and <i>Pax6</i> mRNAs and proteins were analyzed by RT-PCR (<b>C</b>) and WB (<b>D</b>), respectively.</p

Induction of Meis1a/b expressions during neural differentiation of RA-primed P19 cells.

<p>Aggregated P19 cells were treated with 5×10⁻⁷ M RA for various times and the expression levels of Meis1a/b mRNAs and proteins were analyzed by RT-PCR and WB with the anti-Meis1 antibody, respectively. (<b>A</b>) Expression patterns of <i>Meis1a</i> and <i>Meis1b</i> mRNAs during neural differentiation. (<b>B</b>) Quantification of the expression levels of <i>Meis1a</i> and <i>Meis1b</i> mRNAs indicated in <b>A</b>. (<b>C</b>) Expression patterns of Meis1a and Meis1b proteins during neural differentiation. (<b>D</b>) Quantification of the expression levels of Meis1a and Meis1b proteins indicated in <b>C</b>. (<b>E</b>) Expression levels of <i>Meis1a/b</i> mRNAs in mouse fetal brain. Total RNAs from the developing brain were analyzed by Northern blotting. (<b>F</b>) Detailed expression patterns of <i>Meis1a</i> and <i>Oct4</i> mRNAs during neural differentiation. (<b>G</b>) Quantification of the expression levels of <i>Meis1a</i> and <i>Oct4</i> mRNAs indicated in <b>F</b>. (<b>H</b>) Detailed expression patterns of Meis1a and Oct4 proteins during neural differentiation. (<b>I</b>) Quantification of the expression levels of Meis1a and Oct4 proteins indicated in <b>H</b>. <i>Ribosomal large subunit protein P0</i> mRNA, β-actin and 28S ribosomal RNA were used as internal controls.</p

Oct4 activates Meis1a expression.

<p>Monolayer-cultured P19 cells were transfected with the pcDNA3-EF1-α-<i>Oct4</i> expression vector and after 24 h <i>Meis1a</i> and <i>Oct4</i> mRNAs and proteins were analyzed by RT-PCR (<b>A</b>) and WB (<b>B</b>), respectively. (<b>C</b>) Stimulatory effect of Oct4 on <i>Meis1</i> promoter activity. P19 cells were transfected with <i>Meis1</i>(−926)-Luc and various amounts of pcDNA3-EF1-α-<i>Oct4</i>. After 24 h, luciferase activities and expression levels of Oct4 were analyzed. *<i>p</i><0.001 significantly different from vacant vector introduced control cells. (<b>D</b>) Schematic presentation of <i>Meis1</i> promoter-inserted luciferase reporter vectors. <i>Meis1</i>(−926)-Luc possesses three putative Oct4-BEs. (<b>E</b>) Functional analysis of Oct4-BEs. P19 cells were transfected with indicated <i>Meis1</i>-Luc vectors and pcDNA3-EF1-α-<i>Oct4.</i> After 24 h, luciferase activities were assayed. *<i>p</i><0.001 significantly different from <i>Meis1</i>(−926)-Luc and <i>Meis1</i>(−335)-Luc. <i>n = </i>3. (<b>F</b>) Association of Oct4, HDAC1, AcH3, 5mC and 5hmC with the <i>Meis1</i> promoter. Genomic chromatin fragments from RA-treated aggregation form of P19 cells for 0 and 6 h were immunoprecipitated with the indicated antibodies and then DNAs were extracted. PCR was carried out using the primer set covering the −360 to −67 region of Meis1 promoter, in which 12 CpG sites exist. Aliquots of 10% antibody-untreated DNA samples were used for input DNA. (<b>G</b>) Quantification of the <i>Meis1</i> promoter-bound Oct4, HDAC1, AcH3, 5mC and 5hmC indicated in <b>F</b>.</p

Meis1a suppresses Oct4 expression.

<p>(<b>A</b>) Reduction of Oct4 by ectopic expression of <i>Meis1a</i> RNA. P19 cells were transfected with the vacant vector or pcDNA3-EF1-α-<i>Oct4</i> and after 24 h, these aggregated cells were treated with RA for 0 and 6 h. Thereafter, expression levels of Oct4 and Meis1a proteins were analyzed by WB with anti-Oct4 and anti-Meis1 antibodies. (<b>B</b>) The suppressive effect of Meis1a on <i>Oct4</i> promoter activity. P19 cells were co-transfected with <i>Oct4</i>(−1059)-Luc and various amounts of pcDNA3-EF1-α-<i>Meis1a</i>. After 24 h, luciferase activities and Meis1a expression levels were analyzed. *<i>p</i><0.001 significantly different from vacant vector-introduced cells. <i>n = </i>3. (<b>C</b>) Schematic presentation of <i>Oct4</i> promoter-introduced luciferase reporters. <i>Oct4</i>(−1059)-Luc has four putative Meis1-BEs. (<b>D</b>) Functional analysis of Meis1-BEs. P19 cells were transfected with <i>Oct4</i>(−1059)-Luc, <i>Oct4</i>(−698)-Luc, <i>Oct4</i>(−506)-Luc, or <i>Oct4</i>(−254)-Luc together with 60 ng pcDNA3-EF1-α-<i>Meis1a</i>. After 24 h, luciferase activities were analyzed. *<i>p</i><0.001 significantly different from <i>Oct4</i>(−698)-Luc, <i>Oct4</i>(−506)-Luc, and <i>Oct4</i>(−254) Luc. <i>n = </i>3. (<b>E</b>) Occupation of the <i>Oct4</i> promoter by Meis1a and HDAC1. Aggregated P19 cells were treated with RA for 0 and 6 h. Genomic chromatin fragments were immunoprecipitated with anti-Oct4 and anti-HDAC1 antibodies and DNAs were extracted. PCR was carried out using the primer set covering the −1062 to −778 region of the <i>Oct4</i> promoter. Aliquots of 10% antibody-untreated DNA samples were used for input DNA. (<b>F</b>) Quantification of the <i>Oct4</i> promoter-bound Meis1a and HDAC1 indicated in <b>E</b>. (<b>G</b>) Schematic presentation of the adjacent Meis1-BEs3/4 region. In this region, putative Pbx- and Hox-BEs also existed.</p

Data supporting publication "Reusable Immobilized Iron(II) Nanoparticle Precatalysts for Ligand-Free Kumada Coupling"

Electron microscopy data (SEM and TEM images) of Fe-based precatalysts developed in Japan for ligand-free Kumada coupling chemistry