Objective To isolate hepatic stem cells from adult rats,and to examine the process and morphologic changes of isolated hepatic stem cells differentiated into hepatic cells and bile duct epithelial cells under in vitro culturing conditions.Methods AAF/PH was used to stimulate the oval cells.Selective enzymatic digestion and density gradient centrifugation were employed to isolate and purify these cells from heterogeneous liver cell population.The hepatic stem cells were induced to proliferate and differentiate into hepatic parenchymal cells by the addition of dexamethasone,DMSO and several growth factors.The morphologic changes of hepatic oval cells to that of hepatic cells and bile duct epithelial cells were examined.Immunofluorescence method was used to detect the hepatic stem cells c-kit and the expression of OV6 antigen.The albumin of hepatic stem cells and the CK19 expression were identified by RT-PCR analysis.Results The activity of freshly isolated oval cells were over 90%,with different colors and size.The diameter of oval cells was about 1/4~1/6 that of hepatocytes.Large and round hepatocytes appeared first during differentiation,and the cell morphology changed when the oval cells differentiated to hepatocytes.Freshly isolated oval cells was positive for c-kit and OV6 with immunofluorescence staining,and RT-PCR analysis indicated that the albumin and CK19 mRNA were expressed in the new isolated cells.Conclusion The oval cells could be induced to differentiate into a variety of cell lineages including hepatocytes and bile duct epithelial cells in the presence of DMSO and several growth factors.Several antigens such as c-kit,OV6,CK19 and albumin may be expressed in the freshly isolated hepatic stem cells,and the course of morphologic changes may be observed under in vitro conditions of induction and differentiation.

Objective To study the expression and cellular distribution of?-catenin in hepatocellular carcinoma and in HepG2 and SMCC-7721 cells.Methods Immunohistochemistry,immunocytochemistry techniques and Western blotting analysis were used to examine the expression and distribution of?-catenin in the cell membrane,cytoplasm and nucleus of bepatocellular carcinoma tissues,HepG2 and SMCC-7721 cells.Results Immunohistochemical staining showed that the?-catenin poorly,unevenly,even not distributed on the mem- brane of hepatocellular carcinoma cells.The expression of?-catenin on the membranes of low differentiated liver cancer cells was lower than that in the highly differentiated liver cancer cells,but it was strongly expressed in the cytoplasm of hepatocellular carcinoma cells,The ex- pression of?-catenin in the membranes of hepatocellular carcinoma cells was found to be reduced obviously in 18 out of 20 patients(90%).?-catenin was found to be poorly,unevenly,even not distributed in the membrane cells in 10 patients(50%).An obvious accumulation, but with poor-distribution,of?-catenin was observed in the cytoplasm of 17 cases(85%).Significant differences in the?-catenin expres- sion in both membrane and cytoplasm were found among differently differentiated stages of liver cancer cells.In 7 of 9 cases(77.8%)with highly differentiated liver cancer cells,?-catenin was continuously and evenly distributed in both the membrane and cytoplasm,while in 4 of 5(80%)cases with poorly differentiated liver cancer cells the expression of?-catenin in the membrane was found to be significantly poor and showed a uneven distribution in the cytoplasm.The?-catenin expression was positive in the cytoplasm of 5 of 6(83.3%)cases with moderatdy differentiated liver cancer cells.No expression of?-catenin was found in the membrane of HepG2 and SMCC-7721 cells,while it was highly expressed in both cytoplasm and nucleus.Western blotting showed that the?-catenin expressed in both cytoplasm and nucleus of all the liver cancer tissue,and also in HepG2 and SM(3C-7721 cells.Conclusions The expression of?-catenin was down-regulated in the membrane of hepatocellular carcinoma cells and HepG2 and SMCC-7721 cells,while?-catenin was strongly expressed in cytoplasm,and it was obviously accumulated in the nucleus.The distribution of?-catenin was related to the stage of differentiation of cancer cells.

Objectives:To construct rGA antisense eukaryotic expression vector. Methods:Using molecular cloning technology, we subcloned rGA cDNA in its antisense orientations of the pcDNA 3.0 vector. Results:Cutting with Eco RⅠ and Hin D Ⅲ,antisense recombinant plasmid produce 2 bands. One was 0.28 kb fragment, the other was 5.5 kb fragment which corresponded to theoretical calculation. Conclusions:We succeeded in constructing rGA antisense eukaryotic expression vector.

Objective To construct the recombinant plasmid carrying antisense GA cDNA, and evaluate the effect of antisense GA gene on the biological characteristics of colon cancer cells. Methods The recombinant plasmid pcDNA3.0/GA containing antisense GA gene was constructed by gene recombination technique. The recombinant plasmid was transfected into colon cancer cells by lipofectamine. The effect of antisene GA gene on the biological characteristics of colon cancer cell line DH52 was evaluated by MTT and electron microscopy. Results The recombinant plasmid pcDNA3.0/GA was successfully constructed. The growth rate of colon cancer cells decreased, and microstructure of the cells appeared typical apoptotic changes after transfection. Conclusion Antisense GA gene could inhibit the growth of colon cancer cells possibly by suppressing GA gene expression and decreasing GA activity in the colon cancer cells.

Objective To investigate the influence of composite transplantation of two kinds of acellular xeno-dermis (pigskin) and micro-autograft on wound healing. Methods Seventy-two Sprague-Dawley rats with full-thickness skin defect on the back were randomly divided into three groups, and then compositt skin I(acellular dermal xeno-matrix+autologous particulate skin +selective acellular xeno-dermis) was adopted to cover the skin defect in the experimental group I, composite skin II(acellular dermal xeno-matrix+autologous particulate skin + glutaraldehyde xeno-skin) was adopted to cover the skin defect in the experimental group II, and auto-particulate skin was adopted in the control group. The area of wound healing and the rates of wound contraction were calculated.Tissue samples were harvested and examined by means of histology, immunohistochemistry and electron microscopy. Results There were continuous epidermis and basement membrane, and the dermal matrix was a kind of matrix with normal structure and organization of collagen and without any cellular components in selective acellular xeno-dermis. There was full differentiation of epithelium, orderly collagen arrange and basal membrane of the skin could be identified by immunohistochemical stain in compound skin grafting group I. The healing effect of compound skin grafting group I was better than compound skin grafting group II and auto-particulate skin grafting group. Conclusion The composite grafts constructed by selective acellular xeno-dermis (pigskin) could be a potential new type of composite skin substitute for the repair of full-thickness skin defect.

Objective To explore the role of the sensory neuropeptide in the chronic cough induced by postnasal drip syndrome(PNDs).Methods Patients of PNDs with and without chronic cough who came to the First Affiliated Hospital of Ji Nan University between Mar.2004 and Sep.2005 were enrolled and induced sputum by hypertonic saline aerosol inhalation was performed.Twenty-three cases of PNDs with chronic cough and 16 cases of PNDs without chronic cough were collected.Substance P(SP)and calcitonin gene-related peptide(CGRP)in the supernatant of the induced sputum were measured by radioimmunoassay.Results SP and CGRP in the supernatant of the induced sputum were significantly higher in the patients with chronic cough induced by PNDs[SP(345.14?72.58)mg/L,CGRP(573.78?210.96)mg/L]as compared with those normal subjects[SP(168.14?56.97)mg/L,CGRP(227.69?70.84)mg/L,P

Objective To discuss the feasibility of computerized tomography (CT) calibration technique for the quantification of the hepatic fat content in vivo . Methods Eleven healthy volunteers, 53 patients with fatty liver, 10 cases of chronic hepatitis B and 10 cases of liver cirrhosis were tested with CT calibration test tubes. Test tubes contained solutions with Terbutyl alcohol, which CT scan density (CTD) is close to fat tissue, and 18.5% glucose solution which CTD is almost identical to normal liver. Variable Terbutyl concentrations of 10%, 20%, 30% and 40%, which stands for various fat content in liver, were used to measure patients with variable degrees of steatosis under CT scan. Results There was a negative linear correlation ( r = -0.982 , P

Hepaticbililary surgery interventional therapy is a technique which applied to hepaticbilliary diseases for diagnosis and treatment presently.With the development and widespread of interventional therapy technology,there are increasing demands for interventional doctors.But the interventional therapy with invasive and radioactive makes itself impossible for interns to have much chance of actual practice,which becomes the "bottleneck"of the technology development of teaching.Computer-assisted instruction has become an important approach in medical education.It is possible that multimedia courseware can break through the "bottleneck".So we decide to design and make a kind of multimedia courseware adapted to hepaticbililary surgery intentional therapy to improve quality and efficiency of teaching

Precise verification is required in the execution of intensity-modulated radiotherapy (IMRT) for its complexity. Thereinto, the verification of dose distributions is the emphases and nodus. Some problems in the verification by using radiotherapy planning dose verification system (DVS) are explored such as how to verify the precision and credibility, how to reduce errors in every verification step, how to provide quantified analysis. A suit of practical methods and techniques are summarized from equipment selection to quantified analysis.

Obiective To study the regularity of three-dimensional displacement in respiration of lung cancer. Methods Af-ter CT scanning, the CT images were transferred into target planning system (TPS) to obtain three-dimensional reconstruc-tion images. Then target drawing and noting of target position, tumor volume, lung function, patient gender, age and weight were performed. The images during the course of inspiration and expiration in simulator were scanned and transferred into TPS by DICOM RT port. At last, the three-dimensional displacement curve of the interested points(X,Y and Z axis) could be obtained. Results The three-dimensional real time displacement curve of lung cancer interested points could be estab-fished by the way of collecting simulator images and drawing interested points. The maximum displacement range of the interested points were 4.2mm in Z-axis(head-foot direction), 3.8ram in X-axis(left-right direction), 2.9ram in Y-axis(ante-rior-posterior direction). Conclusion The three--dimensional real time displacement curve of lung cancer at any point of the target can be established by collecting the dynamic images in simulator and reconstructing of the corresponding images in target planning system through the DICOM RT.