Issue of individual integration of children with medical disabilities in regular kindergarten

Chronic hypoxic exposure induces the alteration of blood components and skeletal muscle phenotype. Change in blood profile after exposure to 10 % hypoxic condition for 4 weeks. A. Red blood cell count. B. Hemoglobin level. C. Hematocrit value (n = 5–6 per group). D. Mass of soleus, and gastrocnemius of male under normoxia (Nx), normoxia treated with FK-506, hypoxia (Hx), and hypoxia treated with FK-506 at 8–10 weeks of age normalized to body weight (n = 5–6 mice per group). E. Appearance of skeletal muscles, including the soleus and gastrocnemius, in normoxia and hypoxia. F. The composition of muscle fiber-type was analyzed by immunostaining. Frozen sections of the soleus and gastrocnemius muscle after exposure to normoxia and hypoxia were stained with antibodies for MyHC I/slow, MyHC IIa and counterstained for laminin (n = 4–6 mice per group). *p < 0.05, **p < 0.01 compared to control. Values are means ± SEM. (PDF 5163 kb

Additional file 3: Figure S4. of Prolyl hydroxylase domain 2 deficiency promotes skeletal muscle fiber-type transition via a calcineurin/NFATc1-dependent pathway

The expression of genes related to ubiquitin proteasome system in skeletal muscle. The mRNA level of Murf-1 and antrogin-1 in gastrocnemius muscle (n = 4 per group). Values are means ± SEM. (PDF 58 kb

Histological analysis.

<p>Representative photomicrographs of control, vehicle, TM5484 (5 mg/kg/d) and fingolimod (0.5 mg/kg/d) sections at dpi. 28. Spinal cord was sectioned and stained with Cd11b (A), MBP (B), SMI-32 (C) and Collagen IV (D). Four sections of the spinal cord (lumbar spinal segments) from each group were examined. Data are expressed as the mean and corresponding SEM. TM5484 significantly reduced macrophage infiltration/microglia activation, demyelination, axonal degeneration as well as deposition of collagen IV in the spinal cord.</p

<i>In vitro</i> BBB penetration assay.

<p>Analysis of the penetration ratio of different compounds using an <i>in vitro</i> model of BBB. A permeability coefficient (Papp) above 20 x 10^–6 cm/s is considered to provide a very good penetration into the CNS, while a Papp lower than 10 x 10^–6 cm/s is consistant with a low penetrant CNS drug. TM5484 has a 67.6 x 10^–6 cm/s permeability coefficient. Verapamil (N.D.) and Propanolol (31.3 x 10^–6 cm/s) were used as negative and positive control, respectively. Data are mean±SD. n = 3 for each compound. N.D. indicates not detectable.</p

Clinical score (Preventive and combined therapy).

<p>Mice were injected with myelin oligodendrocyte glycoprotein^35-55 peptide in an equal volume of complete Freund’s adjuvant. A, Administration of TM5484 (5 mg/kg/d) from day 0 not only abolished paralysis but also delayed the onset of symptoms in EAE mice. **<i>P</i><0.01 or ***<i>P</i><0.001 Vehicle <i>vs</i> TM5484, †<i>P</i> < 0.01, ‡<i>P</i> < 0.001 Vehicle <i>vs</i> Fingolimod by 2-way ANOVA and Bonferroni test, n = 8–10. B, Simultaneous administration of TM5484 (10 mg/kg/d) with fingolimod (0.5 mg/kg/d) attenuates the clinical score to a significantly greater extent than either TM5484 or fingolimod alone. Data are shown as the means and corresponding SEM. *<i>P</i><0.05 or ***<i>P</i><0.001 Fingolimod <i>vs</i> TM5484 + Fingolimod, by 2-way ANOVA and Bonferroni test, n = 8–10.</p

Clinical score in the rat MS model.

<p>Rats were injected with myelin basic protein^69-88 peptide in an equal volume of complete Freund’s adjuvant. Administration of TM5484 (5 mg/kg/d) from the onset of symptoms significantly attenuated the paralysis score in EAE-treated rats (P<0.001 vs. vehicle at dpi. 15). Fingolimod (0.5 mg/kg) also reduced the paralysis score to a level similar to TM5484 (P<0.001 vs. vehicle at dpi. 15). The peak of the disease was selected to analyze the paralysis score as all animals, including vehicle, had an almost complete recovery by day 22. Data are shown as the means and corresponding SD. *<i>P</i><0.05 or ***<i>P</i><0.001 Vehicle <i>vs</i> TM5484, †<i>P</i> < 0.01, ‡<i>P</i> < 0.001 Vehicle <i>vs</i> Fingolimod by 2-way ANOVA and Bonferroni test, n = 6–8.</p

Gene expressions.

<p>Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of Tnf-1, PAI-1, Cd68, Ccl2, il-6 and BDNF in spinal cord. EAE mice show an increased expression of proinflammatory cytokines as well as a decreased expression of BDNF in spinal cord. Fingolimod has no effects on expressions of PAI-1 and BDNF. Expression levels of all markers are normalized to b-actin. Data are shown as the means and corresponding SEM. **<i>P</i><0.01 by 1-way ANOVA and Dunnett test, n = 6–7</p