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.

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

BACKGROUND/AIMS: There is a controversy about the availability of invasive treatment for esophageal/gastric varices in patients with Child-Pugh class C (CP-C) end-stage liver cirrhosis (LC). We have evaluated the validity of invasive treatment with CP-C end-stage LC patients. METHODS: The study enrolled 51 patients with CP-C end-stage LC who had undergone invasive treatment. The treatment modalities included endoscopic variceal ligation in 22 patients, endoscopic injection sclerotherapy in 17 patients, and balloon-occluded retrograde transvenous obliteration (BRTO) in 12 patients. We have investigated the overall survival (OS) rates and risk factors that contributed to death within one year after treatment. RESULTS: The OS rate in all patients at one, three, and five years was 72.6%, 30.2%, and 15.1%, respectively. The OS rate in patients who received endoscopic treatment and the BRTO group at one, three, and five years was 67.6%, 28.2% and 14.1% and 90.0%, 36.0% and 18.0%, respectively. The average of Child-Pugh scores (CPS) from before treatment to one month after variceal treatment significantly improved from 10.53 to 10.02 (P=0.003). Three significant factors that contributed to death within one year after treatment included the presence of bleeding varices, high CPS (≥11), and high serum total bilirubin levels (≥4.0 mg/dL). CONCLUSIONS: The study demonstrated that patients with a CPS of up to 10 and less than 4.0 mg/dL of serum total bilirubin levels may not have a negative impact on prognosis after invasive treatment for esophageal/gastric varices despite their CP-C end-stage LC.
Balloon Occlusion ; Bilirubin ; Endoscopy ; Esophageal and Gastric Varices ; Hemorrhage ; Humans ; Hypertension, Portal ; Ligation ; Liver Cirrhosis ; Liver ; Prognosis ; Risk Factors ; Sclerotherapy ; Varicose Veins