Objective To study the osteogenic capability of bone marrow stromal cell (BMSc) using tissue engineering methods Methods The osteogenic potential in vitro of cultured BMSc in a conditional medium were examined by phase contrast microscopy,histochemistry stains technique The BMSc were cultured in combination with bioactive glass ceramic (BGC) materials Then the composite were implanted into the skeletal muscle beds in rabbits All implants were exmined by gross observation and histological examination Results The BMSc showed a similar property to those of osteoblasts and could synthesized mineralized new bone tissue in vitro New bone tissue can be observed in the composites of the BMSc and BGC materials Conclusions New bone tissue can be formed by tissue engineering methods

Objective To Observe the effects of tumor necrosis factor ? (TNF ?) on cell proliferation and Intracellular free calcium concentraction ([Ca 2+ ]i) in endothelium of human umbilical vein endothelial cells (HUVEC) and investigate the pathogenesis of pregnancy induced hypertension syndrome (PIH). Methods Confluent monolayer of HUVEC was directly incubated with TNF ? at following final concentrations: 500, 1 000, 2 000 U/ml for 24 hours. The percentages of different cellcycles and [Ca 2+ ]i were measured by flow cytometry and fluorospectrophotometry. Results Incubated with TNF ?, the endothelial cells were elongated and transformed into fibroblast like cells. Border of nucleus was sharp, clarity, and cells were in regular shape. But there were abnormal granules in cytoplasma and some cells detached at the concentrotion of 2 000 U/ml of TNF ?. Stimulated by TNF ?, the percentage of cellcycles from phase G 0+G 1 to S and G 2+M decreased significantly and it was dose dependent. [Ca 2+ ]i increased significantly and dose dependent as well. Conclusion TNF ? may injure endothelium directly and inhibit its proliferation and repair through the changes of [Ca 2+ ]i level. It may play an important role in the pathogenesis of PIH.

Objective　To study the biocompatibility of bioactive glass ceramic (BGC) materials with bone marrow stromal cell (BMSc) and the osteogenic capability of BMSc using tissue-engineering methods. 　Methods　The osteogenic potential in vitro of cultured BMSc in a conditional medium was examined by histochemistry stains technique. The BMSc was cultured in combination with BGC. The attaching and extending speed of the cells to the materials, the proliferation and alkaline phosphatase activity were tested. Then the composite was implanted into the skeletal muscle beds in rabbits. All implants were examined by gross observation and histological examination. 　Results 　The BMSc showed a similar property to those of osteoblasts. BMSc can attach to and extend on BGC materials. No inhibition to celluar proliferation and ALP activity were observed by the materials. New bone can be observed in the composites of the BMSc and BGC materials.　 Conclusions　 BMSc may provide a rich cellular resource in tissue-engineered bone formation. New bone tissue can be formed by tissue engineering methods.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Cyclophosphamide ; therapeutic use ; Dexamethasone ; therapeutic use ; Disease-Free Survival ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; L-Lactate Dehydrogenase ; blood ; Lymphoma, T-Cell ; blood ; drug therapy ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Remission Induction ; Retrospective Studies ; Survival Rate ; Transplantation, Autologous ; Vincristine ; therapeutic use ; Young Adult

BACKGROUND: It is a key point to revascularize the tissue-engineered bone during the repairing of large bone defect. Fascia flap is commonly used in clinic to accelerate the blood supply of implant.OBJECTIVE: To observe the feasibility of repairing goat tibia defects with tissue-engineered bone and accelerating revascularization with fascia flaps.DESIGN: Randomized and controlled animal experiment SETTING: Department of Traumatic Orthopaedics, Nanfang Hospital,Southern Medical University.MATERIALS: Totally 36 goats with the body mass of 14.5-15.5 kg of either gender were enrolled.METHODS: This experiment was conducted at the Department of Traumatic Orthopaedics, Nanfang Hospital, formerly the First Military Medical University of Chinese PLA from December 1999 and December 2003.Bone and periosteum defects 20 mm long were made and fixed with plate of left tibia in 36 goats. They were randomly divided into four groups: Group A in which the defects were filled with coral hydroxyapatite (CHAP), Group B I CHAP+ bone marrow stroma cells (BMSc); Group C with fascia flaps;Group D with nothing. Next, the bone regeneration and the revasculariza tion were evaluated. Radionuclide bone imaging was done 2, 4, 8 weeks after operation. After X-ray examination, the index of optical density of Xray films and histology of the implants were analyzed at 4, 8, 12 weeks after operation, and the biomechanical characters were studied 12 weeks postoperatively.MAIN OUTCOME MEASURES: Gross observation and X-ray, radionuclide bone imaging, biomechanical and histological observation RESULTS: Totally 36 goats entered the stage of result analysis. ① Gross observation of the repair sample of bone defects of the animals in each group: there was no osteogenesis postoperatively at each time point in the blank control group . In Group B, at week 8 to 12, there was no obvious osteogenesis and callus formation on the surface of the materials. In Group C,At weeks 8 to 12, bone defects were filled gradually, many bone callus processes were seen on the surface of the materials , centralizing and enwrapping the materials. The osteogenetic process in the Group C were superior to that of theGroup B. ②Examination result with -901/SA PET-CT scanners: It was seen by naked eyes that at weeks 2 to 8 in the Group A,the radioactivity concentration at region of interesting (ROI) of the operation side had obvious increasing trend, and similar trend of changing appeared in the Group B and Group C, but the ROI counts and T/NT value in the Group B were both lower than those in the Group C. The decreasing trend in the Group A was lower than that in the Group B. ③) Radiological results: the osteogenesis volume through measuring absorbance in the order from large to small was Group C, Group B, and Group A[At week 12, they were (4.180±0.192), (3.480±0.453), (2.959±0.682)respectively ].④Biomechanical results: there were significant difference of loading and bending stress in the Group C, Group B and Group A [ The loading was (758.333±88.754), (530.214±65.297), (359.667±60.715)N , respectively; and the bending stress was (13.937±2.199), (10.123±1.243),(6.223±0.945)N/mm2, respectively ].⑤)Histological results: Slices at various time points in the blank control group showed no bone tissue. In the other three 3 groups, with the prolongation of time, the osteagenetic range and quality were in the order of Group C, Group B and Group A.CONCLUSION: The fascia flaps can accelerate the revascularization process in the formation of tissue-engineered bone so that the capability of tissue engineered bone to repair the large bone defects may be enhanced.

BACKGROUND: The main aspect of the study in the bone histological engineering is how to maintain and improve theosteogenesis of the osteoblasts in vivo and in vitro. The gene transference may provide a new effective method to deal with theproblem.OBJECTIVE: To discuss the effect of the reverse transcription virus mediated human bone morphogenetic protein7(hBMP-7) gene transfection on the proliferation and osteogenetic differentiation of the bone marrow mesenehymal stemcells (BMSCs) of the rabbits.DESIGN:Cells taken as the study object, grouping control, repeat observation andmeasurement.SETTING: Traumatological and othopaedic lab of a medical university hospital.PARTICIPANTS: The study wascompleted in the Traumatological and Othopaedic Lab in the Affiliated Nanfang Hospital of the Southern Medical University from July 2001 to July 2003. Four New Zealand rabbits,whose weights varied from 1.0 to 1.5 kg, were provided without sexlimit by the Animal Experiment Center of the First Military Medical University of Chinese PLA.METHODS:The reverse transcription virus carriersof the hBMP-7 were constructed,and then the BMSCs were transfected by the virus containing target genes. The expression of the hBMP-7 protein was detected with the immunohistochemical method. The cell proliferation, cycle and ALP synthesis were respectively detected with the MTT method,flow cytometer and NPP method.MAIN OUTCOME MEASURES: Primary results: ① the detection results of the cell proliferation. ② the detection results of the ALP.Secondary results: ① the expression of the hBMP-7 protein in the transfected BMSCs. ② the detection results of the cell cycle.RESULTS: After the BMP-7 gene transfection, there was hBMP-7 positive expression in the BMSCs of the rabbits,using the immunohistochemical detection. There was no significant change in the BMSCs proliferation of the rabbits after the hBMP-7 gene transfection ( P ＞ 0.05). Compared with the ALP synthesis of the transfected BMSCs(294. 592 ± 86. 567) nkat/L, there was significant difference in the ALP synthesis of the empty carrier transfected BMSCs(155. 231 ±86.567) nkat/L and the un-transfected BMSCs (160. 866 ±91. 585)nkat/L( F =5. 660, P ＜ 0. 05).CONCLUSION: After the BMP-7 gene transfection, the BMSCs can synthesize and express the extragenous BMP-7. The hBMP gene transfection can promote the differentiation of the BMSCs cultured in vitro into the osteoblasts and can be used as the seed cells in the construction of the histological en gineering bone tissues and in further application.

Objective To investigate the influence of heme oxygenase-1 (HO-1) on rat renal tubular epithelial cell apoptosis induced by albumin and the possible mechanism. Methods The renal tubular epithelial cells (NRK-52E) were cultured in DMEM/F12 1:1 medium as normal control group; NRK-52E cells were cultured with 30 g/L fat-free bovine serum albumin (BSA) as the BSA control group; NRK-52E cells were cultured with CoPP (Cobalt pretoporphyrin Ⅸ) 5 μ mol/L for 24 hours as the treatment group. MTT assay was used to observe the effects of CoPP on growth inhibition induced by BSA in NRK-52E cells. The effect of CoPP was observed in BSAinduced apoptosis with the fluorescence microscope dyed by AnnexinV-FITC PI. The levels of HO-1, and expression of Bcl-2 and Bax mRNA were detected by reverse transcript polymerase chain reaction (RT-PCR). Results Compared with normal control group, BSA inhibited the growth of NRK-52E cells (P<0.05) and increased cell apoptosis rate (P<0.05). The CoPP pretreatment partially inhibited the BSA-induced apoptosis(P<0.05). Compared with normal control group, HO-1 mRNA expression increased (0.44±0.06 vs 0.39±0.05, P<0.05) in BSA control group. Compared with the BSA control group, the expression of HO-1 mRNA significantly increased after CoPP pretreatment(0.50±0.06 vs 0.44±0.06, P<0.05 ). Meanwhile, BSA increased the expression of Bax mRNA (0.87±0.04 vs 0.67±0.03, P<0.05)and reduced the expression of Bcl-2 mRNA (0.25± 0.04 vs 0.42±0.02, P<0.05 ). CoPP could inhibit the effect of BSA (Bax mRNA: 0.75±0.07, Bcl-2 mRNA: 0.36±0.03, P<0.05, respectively). Conclusions BSA can increase the apoptosis rate significantly and regulate the expression of apoptosis associated proteins in mRNA level directly. CoPP inhibits these changes, which provides evidence to support the essential role of HO-1 in cytoprotective function .

Objective: To study the protective effects and the mechanism of Salvia yunnanensis extract on hypoxia/reoxygenation injury in cultured rat H9c2 cardiomyocytes.Methods: The hypoxia/reoxygen (H/R) injury model was established in H9c2 cell strain with or without the extract of Salvia yunnanensis.The cultured H9c2 cardiomyocytes were randomly divided into 6 groups: the normal control (C) group, H/R group, H/R+verapamil (H/R+V) group, H/R+Salvia yunnanensis extract at low dose (H/R+L, 0.01 mg·L-1) group, medium dose (H/R+M, 0.1 mg·L-1) group and high dose (H/R+H, 1.0 mg·L-1) group.The cell viability was measured by MTT assay and the activity of malondialdehyde (MDA) and lactate dehydrogenase (LDH) was measured by a detection kit.Fluorescence absorbance (A) value was measured by a fluoroscopy to show the intracellular reactive oxygen species (ROS) levels.Results: Compared with that in the model group, the survival rate of myocardial cells was significantly higher in Salvia yunnanensis extract at low, medium and high dose groups (P＜0.05 or P＜0.01), and the intracellular LDH leakage (P＜0.05 or P＜0.01), the content of MDA in cytoplasm (P＜0.01) and the intracellular ROS levels significantly decreased in Salvia yunnanensis extract at high dose group (P＜0.05).Conclusion: The extract of Salvia yunnanensis has protective effect on hypoxia/reoxygenation injury in cultured rat H9c2 cardiomyocytes, and the mechanism may be related to the reduction of lipid peroxides and removal of cell oxygen free radicals.

Hepatitis, Autoimmune ; Humans ; Immunoglobulin G

Objective To establish a network laboratory for blood cell analysis and better calibrate haematology analyzers in local lab.Methods According to GB/T 15481《General requirements for the competence testing and calibration laboratories》(idt ISO/IEC 17025),we established a network laboratory providing traceability for blood cell analysis.Complete blood count was traced to Calibration Laboratory in NCCL;The secondary standard haematology analyzer with the same model and calibrator with same lot number were used for verification for a long period.Fresh blood from healthy people was used to calibrate haematology analyzers.Results Gradually we have improved our laboratory quality management system, precision as well as accuracy,which was satisfactory.The unified blood sample was adopted to calibrate different equipments in our hospital and showed consistence when compared with calibration analyzer.The correlation coefficient of all tests is more than 0.99.The relative deviation of WBC,RBC,HCT,HGB and PLT are within?7%,?3.5%,?4%,?3% and?15%,respectively.Conclusions Secondary standard systems provides good comparable results with calibration laboratory.Its tracing mode and quality control scheme could ensure the traceability and accuracy of completed blood count.Furthermore,using elective fresh blood from healthy people,the comparable results from different analyzers were achievable.