Humans ; Mycoplasma pneumoniae ; Pneumonia, Mycoplasma ; diagnosis

Objective To examine the effect and mechanism of sodium ferulate inhibiting P38 MAPK in macrophage on neurotoxicity which was mediated by fribrilar-amyloid-induced.Methods The isolated peritoneal macropohages were cultured.P38 MAPK protein in nuclear extracts was analyzed by Western blot and production of tumor necrosis factor-?(TNF-?) was detected by ELISA.For neuron-macrophage co-cultures,Microtubute-associated protein-2(MAP-2) expression was detected by immunocytochemical technique.The level of LDH in the medium was measured.Results The production of TNF-? and P38 MAPK protein in nuclear extracts increased significantly by incubation with A?25-35.LDH efflux in neuron-macrophage co-culture medium increased and MAP-2 expression decreased significantly(P

The methods, present research situation, existing problems, and application foreground of multidimensional reconstruction of ultras onic heart images are discussed in the present paper. According to the procedure of multidimensional reconstruction of ultrasonic heart images, the discussions are presented in respect toimage acquisition, processing, and display. Based on the present situation and existing problems, the future development and applica tion foreground are proposed.

10.3969/j.issn.2095-4344.2013.25.010

BACKGROUND: How to deal with bone defect is a big problem to surgeons. In recent years, the development in the technology of molecular biology and tissue engineering provides broad prospect for the clinical treatment of bone defect, which is one of the important study directions in department of orthopedics. The transforming growth factor-beta 1(TGF-β1),one of the important factors in bone formation, plays an important role in bone metabolism and recovery.OBJECTIVE: To observe the therapeutic effects of bone defects with osteoblasts transfected . By TGF-β1 combining with biomimetic biodegradable polymer scaffolds.DESIGN: Randomized controlled trial.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Siderophilin, trypsin, 3H-proline and sirius red, etc.MATERIALS: The experiment was completed in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from March to August in 2003. Twenty healthy adult Sprague-Dawley(SD) rats of SPF grade, weighing 200-250 g, were provided by the Experimental Animal Center of Tongji Medical College.METHODS: The osteoblasts transfected by TGF-β1 gene, combining with poly-DL-lactic acid scaffolds modified with poly-L-lysine, were transplanted into rat tibia defect. Radiographs and histological analysis were performed to evaluate the repair effects.MAIN OUTCOME MEASURES: The X-ray evaluation and histology observation were performed at the 4th and 8th weeks after the operation.RESULTS: Totally 20 SD rats were included in result analysis without one rat missing. ①In the experiment group, X-ray image indicated callus formation, while histology observation showed osteoid tissue and new bone formation, and osteoblasts attached to the surface of the materials after 4 weeks. Eight weeks later, the defect was essentially repaired, and the bone density of new bone was similar to that of the autogenous bone. ②In the control group, there was no formation of callus and osteoid tissue, and few osteoblasts attached to the surface of the materials, and a lot of lymphocytes infiltrated and blood capillary grew in the lacune of materials after 4 weeks. Eight weeks later, the imbedded materials were substituted mostly by fibrous tissue.CONCLUSION: The ideal repair effect of bone defect can be obtained through the combination of molecular biology with tissue engineering.

BACKGROUND:With increased age,bone marrow mesenchymal stem cells(BMSCs)are influenced with regard to quantity and quality,which will induce great damages to the donors.Many studies have focused on seeking substitute MSC source.In contrast it remains controversial whether umbilical cord blood contains MSCs.OBJECTIVE:To isolate MSCs from human umbilical cord blood,and to detect their biological properties.METHODS:Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells.DMEM medium containing fetal bovine serum,penicillin,streptomycin and L-glutamine was added.Following several adherences and purification,the floating cells were discarded.Thus,many adherent cells with a confluence were collected.When cells were 60% 80% confluent,cells were digested by trypsin for subculture.Cells at passages,1,5 and 9 were obtained and their morphological changes were observed.Cell surface antigens were measured using flow cytometry.Growth curves were drawn,and cell viability was determined utilizing MTT.RESULTS AND CONCLUSION:Isolated umbilical cord blood MSCs presented an even size,showing spindle or star-shape fibroblasts-like cells.Umbilical cord blood MSCs at 1,5,9 passages were greatly positive for CD29,CD105 and CD166,but weakly positive for CD34 and CD45.Following 5 days of incubation,cells entered logarithmic growth phase.The number was decreased at day 9.Population doubling time was(53.5±8.32)hours.Cells grew well.Cells at 1-7 passages showed similar viability(P ＞ 0.05).Till passage 9,cell proliferation viability was decreased,but no significant difference was determined(P ＞0.05).Results verified that MSCs can be successfully isolated from umbilical cord blood in vitro.Cells at passages 1-9 presented a good reproductive activity.

Objective To develop a kind of new machine for making animal model for pressure ulcer,and inspect its effect through experiments,in order to lay the foundation for the research of pressure ulcer experiments on animals.Methods This study developed the machine after reviewing the domestic and foreign literature,making full use of the existing experimental platform of our university.Then 55 Sprague Dawley(SD) rats were selected,after anesthesia and the skin preparation,the researchers imposed certain pressure with 70 mmHg/cm2 (1 mmHg=0.133 kPa) on the skin and muscle tissue on the inner left thigh of SD rats by using self-designed machine,pressing for 2 h,then reperfusion for 30 min,3 times a day,a total of 7 days.Results The authors developed a kind of new machine for making animal model for pressure ulcer,and successfully prepared Ⅲ phase pressure ulcers model in SD rats with success rate of 98.2％(54/55).Conclusion This machine can prepare Ⅲ phase pressure ulcers model on animals,it's easy to use and efficient,it can be used for researches in the field of prevention and cure of pressure ulcers.

BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants.
OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.
METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.
RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.

Objective To explore the safety of the human bone marrow‐derived mesenchymal stem cells (hBMSCs) after silencing of human leukocyte antigen A2 expression .Methods We divided the cells into three groups:normal cultured cells of the 8th passage served as control group , and hBMSCs after HLA A2 silencing expression of the 5th and 15th passage as experimental groups 1 and 2 ,respectively .The hBMSCs were recultured by sterile methods .The growth curve ,telomerase activation ,and expressions of P27 ,cyclin D2 and cyclin‐dependent kinase 4 (CDK4) were utilized to explore the safety of the hBMSCs induced by LV‐siRNA‐HLA A2 .The BMSCs were transplanted to the subcutaneous layer of nude mice .Tissue types were detected 24 weeks after transplantation . Results The cell curves had no obvious left or right shift in all the groups . The telomerease activation in experimental groups 1 and 2 did not significantly differ from those in control group . The expressions of anti‐oncogene P27 ,cyclin D2 and CDK4 had no obvious difference between the two experimental groups and control group , either . There was only ectopic osteogenesis 24 weeks after the BMSCs (HLA A2 gene silenced ) were transplanted to the subcutaneous layer of the nude mice .Conclusion There was no obvious evidence to support that hBMSCs had undergone change in safety after the silencing of HLA A 2 expression .

Objective To determine the effects of ?-lipoic acid on bone metabolism in lead-poisoned rats.Methods Totally 31 SD rats were randomly divided into two groups,8 rats in blank control group and 23 in lead group that received gastric lavage with lead acetate(230 mg/L).After 40 days,3 rats from each group were taken for the measurement of lead in the blood and bone.The rats in lead group were then randomly divided into four groups: lead control group and three lead groups with different concentrations of ?-lipoic acid [30,60 and 100 mg/(kg?d)],with 5 rats in each.At the end of the experiment(80 d),the rats were killed and the samples of whole blood,serum and bone were collected to detect osteocalcin content with radioimmunoassay and alkaline phosphatase expression with immunohistochemistry staining.Results ① Blood and bone lead contents in each ?-lipoic treatment group were significantly lower than those in lead control group(P