1.Isolation, culture and differentiation of rat hepatic oval cells into hepatocytes in vitro
Gaosu ZHOU ; Rongbin ZHOU ; Dongmei CHEN ; Yuhui NIE ; Guanglong WU ; Hua HAO ; Guanghui YANG ; Mingjun ZHANG ; Side LIU ; Zhenshu ZHANG
Chinese Journal of Tissue Engineering Research 2008;12(25):4957-4961
BACKGROUND: Hepatic oval cells (HOCs) possess the potential of self-renewal, replication, and clone, proliferation and differentiation into mature hepatocytes under a certain condition. HOCs can be used as biomaterial for constructing biological artificial liver in vitro, employed for in vivo transplantation, as well as for tissue engineering as seed cells. HOCs can be widely used for improving clinical treatment of liver diseases. OBJECTIVE: To establish adult Wistar rat models of HOC proliferation, to perform/n vitro isolation and culture of HOCs, and to study the possibility of induction and differentiation of HOCs into hepatucytes. DESIGN: Observational study. SETTING: Institute of Gastroenterology, Nanfang Hospital, Southern Medical University. MATERIALS: Experiments were performed at the Laboratory of Institute of Gastroenterology of Nanfang Hospital from December 2003 to February 2006. Thirty-six healthy male Wistar rats aged 3-4 months (150-200 g) were provided by Experimental Animal Center of Southern Medical University. METHODS: Male Wistar rats were orally fed with ethionine received two-thirds partial hepatectomy (2/3 PH). HOCs were harvested and purified by two-steps perfusion and Percoll density gradient centrifugation, and then cultured in vitro and induced with hepatocyte growth factor (HGF), oncostatin M (OSM) and fibroblast growth factor-4 (FGF4). MAIN OUTCOME MEASURES: Identification and differentiation of HOCs. RESULTS: The concentration of HOCs was about 1.34×108 L-1 in each rat model after in vitro isolation. These cells were round, oval or polygon, about 1/6 1/3 the size of normal hepatocytes. The nucleus-cytoplasm ratio was relatively large. After 2 weeks, clone-like proliferation of HOCs could be observed. Laser scanning confocal microscopy indicated positive expression of stem cells markers Thy-1 and C-kit in cytoplasm and membrane of HOCs. Immunocytochemistry demonstrated positive stem cells marker alpha fetoprotein (AFP) in cytoplasm of HOCs. HOCs can stably passage and its shape gradually changed after inducing with HGF, OSM and FGF4. HOC volume became larger and HOCs lost their ability of sticking to the wall of culture flask. Apparent positive stain of cytoplasm albumin (Alb) was detected 14 days after induction, and the positive ratio increased along with the extension of inducing duration. Results of cytochemistry indicated a brown or black deposit after glucose-6-phosphotase (G-6-P) staining and red particles after periodic acid-Schiff (PAS) staining. CONCLUSION: Adult Wistar rat models of HOC proliferation are replicated by ethionine feeding combined with 2/3 PH. HOCs can be obtained through collagenase perfusion and Percoll density gradient centrifugation. Rat HOCs can be passaged and cultured in vitro. Under a certain condition, HOCs can be induced and differentiated into hepatocytes.
2.18F-FDG PET/CT in vivo Imaging in Examining Inflammation and Apoptosis in the Early-stage Calcification of Atherosclerosis in Rabbits
Jinpeng XU ; Yang LIU ; Maoxiao NIE ; Yuhui ZHANG ; Mingduo ZHANG ; Quanming ZHAO
Chinese Journal of Medical Imaging 2017;25(8):566-571
Purpose 18F-FDG PET/CT,pathological and immunohistochemical analysis are adopted to explore the value of PET/CT in the early-stage calcification examination of atherosclerosis in rabbits and effects of Pioglitazone in treating early-stage calcification.Material and Methods Sixteen New Zealand male rabbits were randomly divided into two groups:Pioglitazone group and control group,witheight rabbits in each group.Atherosclerosis model was established.Rabbits in Pioglitazone group received gavage with Pioglitazone and were raised with high-fat diet for 20 weeks.Blood was drawn to exam high sensitivity C-reactive protein and matrix metalloproteinase-9.PET/CT was used to measure mean standardized uptake value (SUVmean) and maximum standardized uptake value (SUVmax).Rabbit aorta received immunohistochemical,the plaque area,density of macrophage,percentage of calcification area and apoptosis index between the two groups were determined and compared.Results On 20 week,high sensitivity C-reactive protein in Pioglitazone group (4.27±0.43 vs.6.51 ±0.91,P<0.01),matrix metalloproteinase-9 (41.52± 1.99 vs.62.21 ±3.60,P<0.05),SUVmean (0.55±0.18 vs.0.68±0.21,P<0.01)and SUVmax (0.70±0.19 vs.0.82±0.30,P<0.05) were obviously lower than those in control group.Plaque area,density of macrophage,percentage of calcification area and apoptosis index in control group were obviously higher than those in Pioglitazone group.Plaque area of related artery section was positively correlated with SUVmean (r=0.28,P<0.01) and SUVmax (r=0.25,P<0.05).Density of macrophage was positively correlated with SUVmean (r=0.50,P<0.01) and SUVmax (r=0.46,P<0.01).Percentage of calcification area was positively correlated with SUVmean (r=0.50,P<0.01) and SUVmax (r=0.47,P<0.01).Apoptosis index was positively correlated with SUVmean (r=0.61,P<0.01)and SUVmax (r=0.60,P<0.01).Conclusion Inflammation and macrophage apoptosis are of great importance in the early-stage of atherosclerosis.18F-FDG PET/CT imaging can be used to assess minor calcification.Pioglitazone can reduce inflammatory level of atherosclerosis of the experimented animals,inhibiting early-stage calcification.
3.Research progress in medical imaging methods based on magnetoacoustic effect
Yuhui NIE ; Zhipeng LIU ; Tao YIN ; Shunqi ZHANG
International Journal of Biomedical Engineering 2023;46(6):562-568
Electrical property imaging based on the magnetoacoustic effect has the advantages of non-invasiveness, high contrast, and high spatial resolution, making it a promising technique for early disease diagnosis. In this paper, the representative research progress of three magnetoacoustic multi-physical field coupled methods are reviewed, including magnetoacoustic tomography combined with magnetic induction (MAT-MI), magnetoacoustic tomography combined with current injection (MAT-CI), and magneto-acousto-electrical tomography (MAET). Additionally, the future development of these techniques is also discussed.