Table 1.
List of oligonucleotide primers used for RT-PCR.
Figure 1.
Representative CT images of the femur derived from gastrectomized rats and sham-operated rats.
Compared with the sham group (upper column), the cortical bone in the diaphysis is thinner and the cancellous bone in the distal or proximal epiphysis is more scattered in the GX group (lower column).
Table 2.
CT-based bone densitometry of the femur.
Figure 2.
Representative photographs showing horizontal sections of the calvaria and the femoral diaphysis stained with Villanueva bone stain.
In GX (C and D), the bone marrow cavities are larger and the cortical bones are thinner in the calvaria (A and C) and in the femoral diaphysis (B and D) compared with sham-operated rats (A and B). Scale bar = 500 µm.
Figure 3.
Representative photographs showing microscopic images in the femoral distal epiphysis stained with Villanueva bone stain (A and C) and fluorescence microscopic images of the calcein and tetracycline layers in the same focus plane (B and D).
In GX (B) compared with sham (A), the osteoblasts (indicated by blue arrow) are larger in size and increased in number, while the osteoclasts (indicated by red arrow) are also increased in number. The distances between calcein and tetracycline layers as shown by arrow-head were larger in GX (D) compared with sham (B), indicating the mineral apposition rate is increased in GX. Scale bar = 20 µm.
Table 3.
The histomorphometric parameters in the distal epiphysis of the femur.
Table 4.
The biochemical parameters in the serum.
Table 5.
Fold changes in the gene expressions in the bone, liver, kidney, pituitary gland and thyroid.
Figure 4.
Microarray analysis and data mining using IPA-pathway analysis (biological function).
IPA was used to organize the differentially expressed genes into functionally annotated pathways and networks. In biological functions modified significantly by the 1,709 genes whose expression changed by GX, connective tissue development and function (p = 9.67E-10) and skeletal and muscular system development and function (p = 9.67E-10) were listed. A total of 104 genes were included in connective tissue development and function, and 55 genes were in skeletal and muscular system development and function.
Figure 5.
Microarray analysis and data mining using IPA-pathway analysis (canonical signal pathways).
In canonical signal pathways, the role of osteoblasts, osteoclasts and chondrocytes (p = 5.00E-3) was listed. A total of 36 genes were included in this pathway.
Figure 6.
Network analysis to identify novel mechanisms related to the effect of GX.
Twenty-five networks were generated from the 1,709 genes. Eight networks formed hub constructions. Network 21 was the central hub, whose biological functions were tissue development, hematological disease and immunological disease.
Table 6.
Validation of gene expression changes in the bone.