Objective To compare the effect of hepatic oval cells and bone marrow mesenchymal stem cells to treat liver fibrosis in mice. Methods The C57BL/6 mice were fed with 3,5-diethoxycarbonyl-1, 4-dihydro-collidine (DDC) in a standard food at a concentra-tion of 0.1% to activate the oval cells proliferation, then the nonparenchymal cells were separated by in situ perfusion and density gradient centrifugation, and finally Sca-1 antibody was used in conjunction with magnetic activated cell sorting (MACS) to separate the Sca-1 +cells. Other mice were injected subcutaneously with CCl4 to establish experimental liver fibrosis model. Finally, the fibrosis mice were trans-planted with HOC or MSC or saline by spleen injection according to the group they belonged to. Four weeks later, the liver function index, the hydroxyproline and the histopathological change among the groups were used to evaluate the effect of the treatment. Result The liver function index was improved after transplantation. HOC and MSE treatment could reduce both the hydroxyproline level and alleviate the fibro-sis, but HOC seems more effective in the group with continuing CCl4 injection(P < 0.05). When stopping injecting CCl4, the change of fi-brosis was not visible(P >0.05). Conclusion Both hepatic oval cells and bone marrow mesenchymal stem cells can improve the liver function, and alleviate the fibrosis level by transpleen transplantation, but hepatic oval cells were more effective.

especially in the group of ceasing CCl4 injection. Conclusion After transplanted into the liver by spleen injection, hepatic oval cells could plant in the portal area, which improve liver function and alleviate the cirrhosis level.

Objective To observe the effects of Qifuyin on learning and memory ability and expression of somatostatin in hippocampus on Alzheimer’s disease (AD) rats. Methods SD rats were randomly divided into blank control group, sham operation control group, model group, donepezil group and Qifuyin group. The AD model was established by microinjection of A?1-42 into the bilateral hippocampus. The learning and memory ability of rats were evaluated with a series of tasks to find a hidden platform in Morris Water Maze. The expression levels of somatostatin in hippocampus were measured by immunohistochemical SP techniques. Results In the place navigation test, the mean escape latencies of the model group was obviously increased compared with sham operation control group (P

Objective To establish a digital segmentational data set of larynx based on Chinese visible human (CVH). Methods Magnetic lasso and polygon tools of Photoshop were used to segment the small organs and structures of larynx of CVH to establish the segmentational data set of larynx. After conversion of the image format, the segmentational structures were extracted automatically with Thresholding Method and presented 3-D visualized, and then were checked up by its result of 3D reconstruction with Amira 4.1 software. Results Many small structures of larynx were segmentated, such as laryngeal cartilage, laryngeal muscles, vocal cords and so on. Then the segmentational data set of larynx based on CVH was established, which can be used to 3-D reconstruction accurately. Conclusion The segmentational data set of larynx is accurate and integrated, which is helpful to establish the elaborate model of larynx and can provide the method of color image segmentation.

Objective To optimize the parameters of the low-frequency/low-energy ultrasound combined with micro-bubbles in inducing early apoptosis of DU145 cells (an androgen-independent prostatic cancer cells). Methods In our study, the impact of ultrasonic power, micro-bubbles/cell suspension volume rate and irradiation time were investigated. Three levels of each factor were deifned as ultrasonic power (60, 80, 100 mW), micro-bubbles/cell suspension volume rate (10%, 20%, 30%), irradiation time (30, 60, 90 s). According to the three-factor three-level orthogonal design, nine experiments were carried out. The early apoptosis was detected by lfow cytometry. A new experiment was designed with the optimized parameters. Another group without ultrasound irradiation was designed as the control group. Flow cytometry and transmission electron microscope (TEM) were used to detect the early apoptosis. Results In descending order, the inlfuence of these factors on the cell early apoptosis were:ultrasonic power>micro-bubbles/cell suspension volume rate>irradiation time. Moreover, the inlfuence of each factor level were:80 mW>60 mW>100 mW in ultrasonic power, 20%>30%>10%in micro-bubbles/cell suspension volume rate, 60 s>90 s >30 s in irradiation time. The early apoptosis rate of experiment group was 10.41%, while the control group was 0.94%. TEM showed apoptotic cells in the experiment group. Conclusions The optimized parameter of low-frequency/low-energy ultrasound with micro-bubbles in inducing early apoptosis of DU145 cells are ultrasonic power of 80 mW, micro-bubbles/cell suspension volume rate of 20%, and irradiation time of 60 s. With the optimized parameters, the early apoptosis rate of the experiment group has signiifcant higher than the control group.

Objective To investigate the feasibility of low-frequency ultrasound combined with microbubbles improving transfection of enhanced green lfuorescent protein plasmid (pEGFP) to subcutaneously transplanted prostate cancer in nude mice, and to optimize the parameters of intensity. Methods The model of nude mice of subcutaneously transplanted prostate cancer were built. Then they were divided into 2 groups including low-frequency ultrasound group and low-frequency ultrasound combined with microbubbles group, each group with 15 mice. Then 250μl of mixture of saline and pEGFP (50μg) were co-injected in low-frequency ultrasound group and 250μl of mixture of SonVue and pEGFP (50μg) were co-injected in low-frequency ultrasound combined with microbubbles group through tail vein. According to intensity, they were divided into 3 subgroups respectively, including group 1 W/cm2, group 2 W/cm2 and group 3 W/cm2, each group with 5 mice. Ultrasound was applied at 80 kHz input frequency with 50%duty cycle for 10 minutes after pEGFP injection. The tumors were collected on the third day after treatment. The expression of pEGFP in tumor was examined by laser scanning confocal microscope (ISCM) and the average lfuorescence intensity were estimated. At the same time, routine pathological examination was performed. The mean lfuorescence intensity of low frequency ultrasound group and low frequency ultrasound combined with microbubble group were compared with one-way ANOVA and those of any two groups were compared by SNQ-q test. Results TThe mean lfuorescent light in low-frequency ultrasound combined with microbubbles group were 23.75±2.54, 30.25±1.67 and 59.60±2.03, which were obviously stronger than those of low-frequency ultrasound treatment group (14.04±1.35, 14.66±0.98 and 14.32±1.20), the difference was statistically signiifcant (the value of q were 18.26, 14.12 and 13.88, P<0.05). The rate of gene transfection increased along with the enhancement of the ultrasound intensity (the value of q were 15.33, 17.81 and 13.79, P<0.05). Histology analyses performed by HE staining showed that there was no damage to the tumor tissues in all groups, tumor tissues were intact and without infection. Conclusions Low-frequency ultrasound combination with microbubbles can significantly enhance pEGFP transfecting into subcutaneously transplanted prostate cancer in nude mice. In certain range, improving the ultrasound intensity can increase the rate of gene transfection.

BACKGROUND: There is no accepted treatment for liver fibrosis recently. Bone marrow meaenchymal stern cells (BMSCs) used in the treatment of liver fibrosis has been reported as an effectively treatment, but the mechanism is unclear.OBJECTIVE: To study the regulation of hepatic stellate cells mediated by human BMSCs in vitro.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Center for Stem Cells and Tissue Engineering of Sun Yat-sen University and the Central Laboratory of Third Affiliated Hospital of Sun Yat-sen University from June to December 2008.MATERIALS: Human bone marrow masenchymal stem cells were collected from normal youth volunteers; Human hepatic stellate cells and normal liver call line L-O2 were supplied by the Animal Experimental Center of Sun Yat-sen University.METHODS: The purified human BMSCs and hepatic stellate calls were set up in Transwell co-culture system. The incubation density was 2×104cells/well. L-O2 was set up instead of human BMSCs as negative control. Hepatic stellate cells cultured alone served as blank control group. The culture was performed for 72 hours.MAIN OUTCOME MEASURES: Morphology of hepatic stellate cells and results of immunocytochemical staining. Apoptosis of hepatic stellte calls was determined by flow cytometry. Western blot were used to assay the expression of α-actin.RESULTS: Activated hepatic stellate cells presented fiat and thin shape under an inverted microscope. Fat drop was lack in cytoplasm, a -actin located in hepatic stellate calls, with the presence of high tension fibers. Compared with the L-O2 + hepatic stellate cell and hepatic stellate call groups, the apoptotic rate of hepatic stellate cells was significantly increased in the BMSC + hepatic stellate cell group (P < 0.05). α -actin expression was significantly down-regulated.CONCLUSION: Human BMSCs can inhibit activation of hepatic stellate ceils and promote them apoptosis, which may be the anti-hepatic fibrosis mechanism of BMSCs.

Objective To construct the bioartificial liver by bone mesenchymal stem cell (BMSC) and alginate scaffold. Method Alginate scaffold was used as the cell carrier for the cultivation of BMSC and the differentiation from BMSC into hepatic like cells was induced by the cell factors of HGF, EGF and FGF-4 in the scaffold in vitro. The compatibility of the cells and the scaffold was observed by microscopy and the function of the differentiatd cells was tested. The gene of AFP and ALB was detected by RT-PCR. The secretion of ALB and the urea synthesis of the cells were tested by ALB kit and urea kit respectively. The glycogen synthesis and the CK-18 was tested by the glycogen stanning method and the immunofluorescence test. Results BMSC was able to attach, grow and proliferate well in the alginate scaffold, the well compatibility was observed by microscopy. ALB and urea were detected in the cultivating medium, the gene of ALB and AFP was identified by RT-PCR. The glycogen synthesis ability and the expression of CK-18 were induced during the differentiation. Conclusion The three dimensional atginate scaffold exhibited well compatibility with BMSC, BMSC could be differentiated into the hepatic like cell in the scaffold. BMSC and the alginate scaffold could be used to construct the bioartificial liver for the hepatic tissue engineering.

Objective To analyze the short-term and long-term effectiveness of radiofrequency ablation combined with transcatheter arterial chemoembolization (TACE) for the treatment of hepatocellular carcinoma (HCC).Methods The clinical data of 70 HCC patients who had received thermal ablation (group A) done or in combination with TACE (group B) were retrospectively analyzed.Results The rate of intrahepatic distant recurrence in group B (25 cases) was lower than that in group A (45 cases) (X2 =3.845,P =0.046) and the tumor-free survival rate was higher than group A (X2 =5.020,P =O.030).There were no differences in the local tumor progression rate (X2 =0.853,P =0.374) and overall survival (x2 =2.316,P =0.154) between two groups.Incidence of bone marrow suppression in group B was higher than that of group A (X2 =5.642,P =0.042).Major complications didn't occur in any group(X2 =2.016,P =0.183).The costs was higher(t =7.738,P ＜0.001) and the hospital stay was longer (t =5.921,P =0.003) in group B than group A.Conclusions Compared with ablation alone,combined therapy is able to reduce short-term recurrence,and improve tumor-free survival.Combine therapy is safe and effective method for small liver carcinoma.

BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.