Objective We established a model of human breast cancer in nude mice, to discuss the feature of pathology and biology of breast cancer,and give help to establish tools of pathogen research. Methods Human breast cancer cells MCF-7 were subcutaneously injected into the right armpit of nude mice to establish human breast cancer models.The mice were divided into composite group, estrogen group, leptin group and the blank group (30 in each). In the composite group,estrogen and leptin were injected into peripheral region of the tumor daily.In the estrogen group,estrogen was injected.In the leptin group, leptin was injected.In the blank group, physiological saline was injected.Tumor growth was observed, and the volume of the tumor was recorded.The tumor tissues obtained from the mice were pathologically examined. Results (1)The tumor-taking rate of leptin group and the blank group were 46.7% (14/30)and33.3% (10/30) ,and the mode is failure.In composite group and estrogen group they were 96.7% (29/30) and 70% (28/30).There was not significant difference between them ( P < 0.05).(2) The differences of average diameter and volume were statistically significant between the composite group and the estrogen group (P < 0.05).(3) Pathology diagnosis was invasive ductal carcinoma. Conclusions (1) Establishing human breast cancer model in nude mice need the stimulation of estrogen. The tumor-taking rate of nude mice has no relationship with leptin.(2) In the study in vivo, leptin as same as estrogen has stimulating effect on MCF-7 cells proliferation.(3) The transplanted cancer cell partly have the pathology and biology features of the human breast cancer cells and give help to establish tools of pathogen research.

Objective To evaluate the different effects of the treatment of thoracolumbar fractures by pedicle of vertebral arch Tenor system and Harrington device.Methods The clinical data of 35 cases of thoracolumbar fractures were analyzed retrospectively, 23 of them were operated with Harrington device and 12 with Tenor system.Results These cases were followed up for 6 months to 2 years. The fractures treated with Tenor system got satisfied reduction, reached about 96 7% of normal vertebral height. The deformities were completely corrected and complications were significant lower than those derived from Harrington device treatment, and no delayed neurologic damage was found. Most cases got reduction with one to three Frankel grades of neurologic recovery.Conclusion The Tenor pedicle system is more effective in treatment of thoracolumbar fracture than non-pedicle device. It can get satisfied fracture reduction , reliable fixation and less complications.

[Objective]To measure adult normal femur specimen in order to obtain accurate femoral resection and match the distal femoral in total knee arthroplasty.[Method](1)Forty-seven adult normal femur specimens of central Chinese were used for taking digital pictures in both anteroposterior and medialateral position.Five angles were measured through drawing lines of fixed point.(2)The exact position of the entry point was defined by the two wires on the specimens.[Result](1)The five angles reflect distal femoral anatomy features on the frontal plane and sagittal plane.(2)The mean value of distance from the entry point to the center of the intercondylar notch was 6.21 mm and the distance from the entry point to the anterior margin of the femoral posterior cruciate ligament(PCL) insertion is found to be 6.7 mm.[Conclusion](1)The distal one-third of femora was found to be outward bowing on frontal plane and forward bowing on Sagittal plane.Especially,the forward bowing is more remarkable than Outward bowing.So,when the position of intramedullary guide stick is decided by distal femoral anatomy,the axis alignments on frontal and Sagittal planes are both important.(2)During total knee afthroplasty the exact position of entry point may be defined by the Center of the intefcondylar notch and femoral PCL insertion.

BACKGROUND:. Growth factor starts a new epoch of the research and application of wound healing, and the effect of the combination of Chinese tradition medicine and biomedicine in wound healing is still in the discussion.OBJECTIVE: To observe the effect in promoting wound healing of rats with bletilla colloid, a Chinese herbal medicine, as a carrier of exogenous high molecular weight nerve growth factor (HMW-NGF).DESIGN: The single sample observation was adopted in the identification of activities of HMW-NGF; The randomized control experiment with animal as subjects was adopted in the observation of the effect of the HMW-NGF mixed with the bletilla colloid as a carrier in promoting wound healing.SETTING: Department of Orthopaedics, Shenzhen Buji People's Hospital,and Department of Orthopaedics, Tongji Hospital Affiliated to Huazhong University of Science and Technology.MATERIALS: Forty SD rats aged 90-120 days of either gender were employed, with the body mass of 220-280 g; The freeze dried preparation of cattle seminal fluid with HMW-NGF with 1mg each.METHODS: From September 2001 to December 2004, the experiment was carried out in the Department of Orthopaedics in Shenzhen Buji People's Hospital and Central Laboratory of Department of Orthopaedics in Tongji Hospital Affiliated to Huazhong University of Science and Technolcolloid, HMW-NGF, HMW-NGF+ bletilla colloid were added into the serum-free medium (SFM), and the influences of them on the dorsal root lected and divided into 4 groups with 10 rats in each group. The rats of normal control group had no medicine on the wound. In the bletilla colloid group, the HMW-NGF group and the HMW-NGF+bletilla colloid group, incision size 2 cm was made at the back of each rat, and bletilla colloid,HMW-NGF, HMW-NGF+ bletilla colloid were applied respectively on the wound, once per day. The wound area was measured at day 3 and 10 after treatment, and the wound area was calculated by the image analysator. The category and the quantity of the emigrated cells were observed with light microscope, and the healing time of the wound was observed.tion under light microscope.the DRG of chick embryo was cultured, the reaction of evection growth around the ganglion in the HMW-NGF, HMW-NGF+ bletilla colloid with the dilution desity of 1:106-1: 108 was most significant, which was not seen in healing with HMW-NGF + bletilla colloid was significantly increased as compared with that in normal control group, the bletilla colloid group and the The healing time of the wound of the normal control group, the bletilla colloid group and the HMW-NGF group and the HMW-NGF+ bletilla colloid group were (19.5±0.7), (17.3±0.6), ( 16.6±0.7 )and ( 14.9±0.4 ) days respecvation of cut sheets 3 days after treatment: The quantity of polymorphonuclear leucocytes, histoleueocyteand fibroblast in the bletilla colloid group,the HMW-NGF group and the HMW-NGF+ bletilla colloid group were significantly larger than that in the normal control group, and the micrangium could be seen in the HMW-NGF group and the HMW-NGF+ bletilla colloid group. The observation of cut sheets 10 days after treatment: The granulation tissue in the HMW-NGF group and the HMW-NGF+ bletilla colloid group was compacter and had more micrangium as compared with the normal control group and the bletilla colloid group.CONCLUSION: HMW-NGF+ bletilla colloid has significant effect on promoting the healing of the wound at the earlier and later period, and the effect is better than that of bletilla colloid or HMW-NGF.

Objective To study the effects and mechanisms of infrasound with low sound pressure level on focal cerebral ischemia-refusion injury. Methods Focal cerebral ischemia was produced by 2 hours of occlusion of the middle cerebral artery in rats. lnfrasound generated by infrasound 8TM device was used as treatment factor. Thirty-two Sprague-Dawley rats were divided into three groups: sham group (n=8), model group (n=8) and in-frasound group (n=16) , and the infrasound group was subdivided into 20- and 120-min infrasound groups, with 8 rats in each group. Neurological symptoms was assessed at 2 h, 1 d, 3 d and 7 d, respectively. These rats were sacrificed after 7 days of infrasound treatment and their brains were harvested. The number of IGF-1 posive cells of ischemia cortex was counted by using immunohistochemical technique. Results Compared with model group, neurological symptoms of rats in 120-min infrasound group was significantly improved (P<0.05); the number of IGF-1 positive cells of ischemia cortex in 120-min infrasound group increased significantly as compared with that in model group (P<0.01). Conclusion Infrasound with low sound pressure level (120 min/d, 7 d) could exert neuroprotective effect in focal cerebral ischemia injury by increasing expression of IGF-1 in brain.

Objective To examine the neuroprotective effect of pulsed magnetic field in a animal focal cere-bral ischemica-reperfusion injury model. Methods Forty-eight Sprague-Dawley (SD) rats were randomized into 3 groups, a sham-operation group, a model group and a pulsed magnetic field group, with rats 16 in each group. Mid-dle cerebral artery occlusion (MCAO) method was employed to establish the focal cerebral ischemia-reperfusion inju-ry model in rats of model group and pulsed magnetic field group. Rats in sham-operation group was subject to the same operation procedure but not underwent ischemia-reperfusion. The infarction volume, histopathological damage and expressions of IGF-1 in ischemic brain tissue were investigated to evaluate the effect of pulsed magnetic field. Results The infarction volume was reduced, histopathological damage alleviated and expressions of IGF-1 in ische-mic brain tissue elevated in the pulsed magnetic field group as compared against the model group (P<0.05). Con-clusions Pulsed magnetic field might provide neuro-protection against cerebral ischemia-reperfusion injury.

BACKGROUND: Some researches suggest that placenta is regarded as a new source of mesemchymal stem cells (MSCs). Adherent cells derived from placenta tissue have similar morphological characteristics and surface markers to MSCs; meanwhile, they can differentiate into osteoblasts and nerve cells.OBJECTIVE: To find a new source and way for MSCs separation.DESIGN: Observational study.SETTING: The Third People's Hospital of Wuxi.MATERIALS: Placenta was sourced from cesarean in our department of obstetrics and gynecology and provided confirmed consent from the relatives and ethics committee. The main reagents contained culture medium, sABC kit, DAB staining kit, caprine-anti-rat FITC, recombinant human basic fibroblast growth factor (rh-bFGF), rabbit-anti-human glial fibriliary acidic protein (GFAP).METHODS: The experiment was carried out in the Laboratory of Cells & Molecular Biology, the Third People's Hospital of Wuxi from May 2005 to August 2006. Placenta entity tissue was digested, adherent cells were cultured, morphological characteristics were observed, and growth curves of placenta-derived multipotent cells (PDMCs) in various generations were drawn. On the 1st and 7th days, supernatant was derived from cells in primary culture to measure content of β-glycerophosphate disodium (β-HCG) with chemiluminescence technique. In addition, expression of surface antigen and differentiating potency were detected at the same time. At 24 hours (at phase of nerve cells) and 2 weeks (at phase of osteoblasts) after inducible differentiation, cultured cells were dealt with routine immunocytochemical stain, and then they were observed under routine microscope or fluorescence microscope.MAIN OUTCOME MEASURES: ① Morphological indexes and growth curves of PDMCs; ② measurement of β-HCG in supernatant of PDMCs and expression of antigen of PDMCs with flow cytometer; ③ immunohistochemical analysis of PDMCs during and after inducible differentiation.RESULTS: ① Primary culture of PDMCs: After digestion of placenta tissue, a few of adherent cells were obtained and gradually formed thin and flat monolayer cells two weeks later. The monolayer cells grew like whirlpool or cluster. With the increase of cell density, soma was slenderer and slenderer and like fibroblast. ② Growth curves of PDMCs: Growth latency ranged from 2 to 8 days after cell inoculation. During this period, adherence was observed gradually but not obviously amplified. Eight days later, cells entered log growth phase. During this period, proliferation was active and cell process surrounding extended under phase contrast microscope. A lot of MSCs were observed in division phase between two nuclei. In addition, density was increased and cells connected to each other. Within 11-14 days, growth curve gradually entered platform phase. MSCs covered the bottom of bottle, cells slowly expanded, and primary culture stopped. ③ Measurement of β-HCG: Expression of β-HCG was not detected in supernatant at two time points. ④ Characteristics of surface antigen of PDMCs: PDMCs could express CD29, CD44 and CD105, but not CD34, CD45,CD19 and CD106. ⑤ Inducible differentiation of PDMCs: At 24 hours after inducing to nerve cells, form of PDMCs obviously changed, soma rebounded, refraction of nucleus was partially reinforced, structure was similar to dendrite and axis-cylinder, and positive neuro-specific enolase and GFAP were observed after staining.CONCLUSION:Placenta tissue contains PDMCs whose morphological function is similar to MSCs; in addition, placenta is regarded as an effective source of MSCs.

BACKGROUND: Multipotential mesenchymal stem cells (MSCs) can be differentiated into bone, fat, cartilage, muscle and endothelial cell at the specific conditions, so it draws great attentions of the related study.OBJ ECTTVE: To establish the method of in vitro culture and expansion of human umbilical blood-derived MSCs and investigate their influencing factors.DESTGN: Randomized and controlled trials.SETTING: Department of Tissue Engineering and Cell Biology in Wuxi Third People's Hospital.MATERTALS: Seventy cases of human umbilical cord blood (HUCB) of healthy neonates (selected from Parturient Room, Department of Gynecology and Obstetrics, Wuxi Third People's Hospital, with the informed consents of the puerpera and their relatives), 40 mL each; dulbecco's minimal essential medium (DMEM) of low glucose (LG) at the dose of 50 g/L, fetal bovine serum (FBS), mycillin and trypsin (all were from Gibco), FITC-labeled mice anti-human CD29, CD105 and CD106 (Ancell) monoclonal fluorescent antibody, PE-labeled mice anti-human CD34, CD44, CD45, CD19, HLA-DR (Immunotech)monoclonal fluorescent antibody, Ficoll separation medium (Pharmacia).METHODS: The experiment was carried out in Department of Tissue Engineering and Cell Biology in Wuxi Third neonatal cord blood were anticoagulated by 25 u/mL heparin, and 60 cases of isolated HUCB mononuclear cell were precipitated and suspended by cell culture fluid (50 g/L DMEM-LG + 50 g/L FBS + 10 g/L mycillin), while other 10 cases were purified with DMEM of high glucose (100 g/L) to produce adherent layer. Once the cells reached fluences of FBS coating on adhesive rats of MSCs: Some of the purified HUCB MSCs at logarithmic phase were growth curve of MSCs: The growth of cells were observed by phase contrast microscope (OLYMPUS CK40), and were detected by EPICS-ALTRA flow cytometry.FBS coating and without FBS coating at the different time points, and their adhesive rates.RESULTS: All 70 cases of HUCB MSCs were adopted in this study, however, 10 samples failed to obtain the ideal MSCs cord blood MSCs after culture. During 2 weeks of culture, primary cells began to double for 3-4 days and reached conand CD105, but did not express antigens CD34, CD45, CD106 and HLA-DR, which was identical to surface labeling of those without FBS coating (P＜0.01).CONCLUSrON: MSCs in HUCB can be cultured and expanded in vitro, and are regarded as an alternative source of MSCs for experimental and clinical applications. Moreover, high glucose can depress the growth and proliferation of MSCs in HUCB.

Objective To observe dynamically the expression of insulin-like growth factor(IGF-1)in the brain and liver,and to explore changes in IGF-1 levels.Methods The thread method waa used to establish middle cerebral artery occlusion in rats.The expression of IGF-1 on the ischemic side of the cerebral cortex and in the liver wag observed dynamically using immunohistochemical techniques. Results The number of IGF-1-positive cells increased significantly,peaking on the 3rd day in the cortex and the 7th day in the liver.Conclusion The expression of IGF-1 Wag upregulated in the brain after the experimental cerebral ischemia;peripheral humoral regulation of IGF-1 responded more slowly and recovered to some extent at the 7th day after cerebral ischemia.

BACKGROUND: There have been no effective means for clinical treatment of large regions of bone defects.Nano-hydroxyapatite/collagen(nHAC)composite would provide a new pathway for repair of bone defects owing to its similar structure to natural skeleton and better biocompatibility.OBJECTIVE: To investigate the role of nHAC composite co-cultured with bone marrow-derived mesenchymal stem cells(BMSCs)in repair of bone defects.METHODS: Following isolation and culture,human BMSCs were co-cultured with nHAC composite.Gross observation,histological analysis,and electron microscope observation were performed to analyze osteogenesis for repair of bone defects in the clinic.RESULTS AND CONCLUSION: Human nHAC could greatly proliferate in vitro.X-ray photography revealed that bone defects well healed after implantation of nHAC/BMSCs composite.These findings indicate that BMSCs exhibit osteogenic potential and nHAC is a satisfactory scaffold material for construction of tissue-engineered bone.