Objective To find out a proper way to detect green fluorescent protein (GFP). Methods Kidneys, livers and femurs from GFP transgenic mice and C57BL/6J wild type mice were employed for in vivo study.The samples were dehydrated with alcohol and acetone individually before embedding, then frozen, paraffin and resin sections were made for the detection of GFP. C3 P12 cells which derived from calvaria bone cells of GFP transgenic mouse were used for the detection of GFP in vitro. Cells were exposed to alcohol, acetone and PBS after paraformaldehyde fixation. Laser scanning microscopy was employed for GFP detection. Results In frozen sections, both kidney and liver samples which exposed to 4% buffered paraformaldehyde fixation had strong GFP signals, while GFP signal disappeared completely in fresh frozen sections without fixation. Much stronger GFP intensity was found in acetone treated samples than in alcohol treated paraffin sections, but without apparent difference in GFP intensity in acetone and alcohol treated resin samples. Acetone and alcohol made no difference in fixed C3 cells in different time courses. Conclusion Acetone treated paraffin sections are preferable for GFP detection.

Purpose : We verified electromyogram activity during hip flexion under different pelvic rotation positions, investigated the effects of pelvic rotational position and defined the difference between males and females. Subjects : 15 healthy adults (5 male, 10 female) with a mean age of 28.8 years participated in this study. Method : We recorded surface electromyograms of the tensor fasciae latae muscle (TFL), rectus femoris muscle (RF), biceps femoris muscle (BF), semitendinosus muscle (ST), and the bilateral internal oblique muscle (OI) during flexion of the hip joint in a supine position with three different pelvis rotation conditions. Results : Males showed no significant differences at all muscle activity levels. Meanwhile, TFL muscle activities were significantly higher for females in other side rotation of the pelvis than in the pelvis neutral position (p<0.05). Moreover, opposite side of OI muscle was activated significantly highly in both side rotation position (p<0.05). Conclusion : Generally, the transverse diameter of the pelvis in females is longer than that in males. When the lower extremity is elevated in the pelvis rotation positions, the moment of force on the pelvis is thought to be higher in females. Therefore, the stabilizing muscles of the pelvis, like the OI, need to be activated isometrically in females.

OBJECTIVETo detect the expression of osteoclast related factors, tumor necrosis factor-alpha (TNF-alpha), receptor activator of NF-kappaB ligand (RANKL) and tartrate-resistant acid phosphatase (TRAP), in the process of bone remodeling.

METHODS8-week-old male C57BL/6J mice were employed in this study to detect the expression of osteoclast related factors by real time PCR.

RESULTSTNF-alpha, RANKL and TRAP were up regulated in the process of bone remodeling, they reached the peak on day 2, 5 and 10 individually after injury.

CONCLUSIONOsteoclast related factors also participate in bone remodeling, which depends on the delicate balance between bone formation and bone resorption.

Animals ; Bone Resorption ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; Osteoclasts ; Osteogenesis ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Tumor Necrosis Factor-alpha

OBJECTIVEAlkaline phosphatase (ALP) and osteocalcin (OC) are the markers of new bone formation. Quantitative study of ALP and OC in the process of new bone formation helps to understand the ongoing of this cascade and contributes to make diagnosis in clinical treatment.

METHODS8-week-old male C57BL/6J mice and primary osteoblasts from neonatal C57BL/6J mice calvaria were used in this experiment. HE staining, Northern blot and Real Time PCR methods were employed to detect the histological changes and the expression pattern of ALP and OC.

RESULTSIn vivo study showed that after fracture the expressions of both ALP and OC kept on increasing which were peaked on the 10 day, then started decreasing gradually. In vitro study on primary osteoblasts showed that the expressions of ALP and OC reached peak on the 14th day in differentiation culture medium and started decreasing from this time point till the 21st day.

CONCLUSIONThe expression of ALP and OC in the process of new bone formation parallels with the development of osteoblasts, it increases with the differentiation of osteoblasts and becomes decreasing with the maturation of osteoblasts. The reciprocal relationship between the expression pattern of ALP and OC and development of osteoblast helps to maintain homeostasis.

Alkaline Phosphatase ; Animals ; Bone and Bones ; Cell Differentiation ; Male ; Mice ; Mice, Inbred C57BL ; Osteoblasts ; Osteocalcin ; Skull