Additional file 1 of Impaired pulmonary function and associated factors in the elderly with tuberculosis on admission: a preliminary report

Supplementary materials: Table S1. The FEV1/FVC ratio and FEV1(%) between different groups of patients. Table S2. Logistic regression analysis of obstructive ventilatory disorders. Table S3 Logistic regression analysis of restrictive ventilatory disorders

Viability of the cells encapsulated in PEGDA gel.

<p>Representative images of live (green) and dead (red) 4T1 cells encapsulated in PEGDA gels with 5 kPa modulus and cultured in stem cell culture medium for 2 (a), 6 (b) and 12 (c) days. Cells were stained with cAM/EthD for live (green) and dead (red) cell imaging. The insets in (a) to (c) are live/dead images of 4T1 cells in PEGDA gels with 70 kPa modulus after 2, 6, and 12 (day), respectively.</p

Effect of CD44BP on tumorsphere formation and CSC marker expression.

<p>Representative fluorescent images of the tumorsphere size and distribution for 4T1 cells encapsulated in PEGDA gels (1.4×10⁵ cells/ml) conjugated with CD44BP (a, conj CD44BP), conjugated with a scrambled sequence of CD44BP (b, conj s-CD44BP), CD44BP dissolved in the gel (c, dis CD44BP), and s-CD44BP dissolved in the gel (d, dis s-CD44BP) and cultured in the stem cell culture medium for 9 days. Effect of CD44BP on cell number density (e) and tumorsphere number density (f) for 4T1 tumor cells encapsulated in PEGDA hydrogel and incubated in the stem cell culture medium for 9 days. Effect of CD44BP conjugation on CD44 (g), CD24 (h), ABCG2 (i) and SCA1 (j) mRNA marker expression for 4T1 tumor cells encapsulated in PEGDA gel and incubated in the stem cell culture medium for 9 days. RNA levels of the cells were normalized to those at time zero. A star indicates a statistically significant difference (p<0.05) between the test group and “Ctrl”. Two stars indicates a significant difference (p<0.05) between the two CD44BP and s-CD44BP groups within the same form of peptide addition (Dis or Conj). Values are expressed as mean ± SD (n = 3).</p

Effect of CD44BP conjugated to the gel on tumor formation <i>in vivo</i>.

<p>The gel without cell (negative control, light blue), 4T1 tumospheres in suspension (positive control, red), 4T1 cells encapsulated in the gel without CD44BP (green), and 4T1 cells encapsulated in the gel with CD44BP (light blue) were inoculated subcutaneously in syngeneic Balb/C mice. Tumor sizes were measured daily from post-inoculation day 11 (n = 6/group). Tumor growth was not observed in the negative control group (the gel without cell) and the group with 4T1 cells in the gel with CD44BP (the lines for these two groups are overlapped in the figure).</p

CSC population in the cells encapsulated in PEGDA gel.

<p>MCF7 cells were encapsulated in PEGDA gels with 5 kPa modulus and cultured in stem cell culture medium. Cells before encapsulation (a), 3 days (b), 8 days (c) and 11 days (d) after encapsulation were stained with CD44−FITC and CD24-PE antibodies. The population of CD24+, CD44+ and CD44+/CD24− cells was determined by flow cytometry. Flow cytometry was repeated multiple times on each sample to ascertain reproducibility of the results.</p

Sphere formation and the effect of cell type encapsulated in PEGDA gel on the expression of CSC markers.

<p>Representative fluorescent images of the tumorsphere size and distribution for 4T1 (a), MCF7 (b), and MCF10a (c) cells encapsulated in PEGDA gels (1.4×10⁵ cells/ml), and cultured in stem cell culture medium. Encapsulated cells were stained with phalloidin for cytoskeleton (red) and DAPI for nucleus (blue). Effect of cell type on cell number density (d) and tumorsphere diameter (e) for tumor cells encapsulated in PEGDA hydrogel and incubated in stem cell culture medium for 6 or 9 days. The sphere size distribution (f) was determined 9 days after encapsulation. Effect of cell type on CD44 (g), CD24 (h) and ABCG2 (i) mRNA marker expression for tumor cells encapsulated in PEGDA hydrogel and incubated in stem cell culture medium for 6 or 9 days. RNA levels of the cells were normalized to those at time zero. A star indicates a statistically significant difference (p<0.05) between the test group and the groups with different cell type in the same time point (the same diameter range in f). Values are expressed as mean ± SD (n = 3).</p

Peptides and their scrambled sequence.

<p>Peptides and their scrambled sequence.</p

Expression of the markers related to CSC maintenance in cells grown in the gel conjugated with CD44BP, IBP, or FHBP.

<p>Effect of cell binding peptide on N-Cadherin (a), E-Cadherin, (b), integrin α_V (c), and integrin β₃ (D) mRNA marker expression for 4T1 tumor cells encapsulated in PEGDA hydrogel and incubated in the stem cell culture medium for 9 days. Effect of cell binding peptide on vimentin (e) and VEGF (f) protein expression. The protein expression was determined by western blot and quantified with imageJ. Actin was used as the internal control and the protein expressions were normalized to those at time zero. RNA levels of the cells were normalized to those at time zero. A star indicates a statistically significant difference (p<0.05) between the test group and “Ctrl”. Values are expressed as mean ± SD (n = 3).</p

CSC population in cells encapsulated in PEGDA gel conjugated with CD44BP, IBP, or FHBP.

<p>4T1 cells were encapsulated in PEGDA gels with 5 kPa modulus and cultured in stem cell culture medium. Cells before encapsulation (a), 9 days after encapsulation in the gel without peptide (b), 9 days in the gel conjugated with FHBP (c), 9 days in the gel conjugated with IBP (d), and 9 days in the gel conjugated with CD44BP were stained with CD44-FITC and CD24-PE antibodies. The population of CD24+, CD44+ and CD44+/CD24− cells was determined by flow cytometry. Flow cytometry was repeated multiple times on each sample to ascertain reproducibility of the results.</p

MiR-133b Is Down-Regulated in Human Osteosarcoma and Inhibits Osteosarcoma Cells Proliferation, Migration and Invasion, and Promotes Apoptosis

<div><p>MicroRNAs (miRNAs) decrease the expression of specific target oncogenes or tumor suppressor genes and thereby play crucial roles in tumorigenesis and tumor growth. To date, the potential miRNAs regulating osteosarcoma growth and progression are not fully identified yet. In this study, the miRNA microarray assay and hierarchical clustering analysis were performed in human osteosarcoma samples. In comparison with normal human skeletal muscle, 43 miRNAs were significantly differentially expressed in human osteosarcomas (fold change ≥2 and <i>p</i>≤0.05). Among these miRNAs, miR-133a and miR-133b expression was decreased by 135 folds and 47 folds respectively and the decreased expression was confirmed in both frozen and paraffin-embedded osteosarcoma samples. The miR-133b precursor expression vector was then transfected into osteosarcoma cell lines U2-OS and MG-63, and the stable transfectants were selected by puromycin. We found that stable over-expression of miR-133b in osteosarcoma cell lines U2-OS and MG-63 inhibited cell proliferation, invasion and migration, and induced apoptosis. Further, over-expression of miR-133b decreased the expression of predicted target genes BCL2L2, MCL-1, IGF1R and MET, as well as the expression of phospho-Akt and FAK. This study provides a new insight into miRNAs dysregulation in osteosarcoma, and indicates that miR-133b may play as a tumor suppressor gene in osteosarcoma.</p></div