Objective:To dynamically observe the change of intra-articular pressure of the hip by means of injecting 2‰Noradrenaline(NA)into the hip joint cavity,to study the effects of intra-articular pressure change on transient synovitis of hip(TSH).Methods:Fourty-eight dogs aged 2~3 months had been randomly divided into 12 groups,with four in each group All were injected 2‰NA into the hip joint an four assessments had been done.①The intra-articular pressure of hip was determined at different times.②99m TC-MDP triphasic imaging was used to observe the local blood supply of femoral head.③Pathological changes were observed to understand the disease course.④X-ray was adopted to exmine the hip.Result:Noradrenaline induced transient synovitis of hip,and synovitis resulted in the increase of intra-articular pressure of hip which was over 20mmHg and lowered to be normal with the inflammation fading away.Transient blood insufficiency was observed in femoral head,but with no putrescence.Conclusion:In transient synovitis of hip,synovitis resulted in the change of intra-articular pressure hip,which changed to be normal as the inflammation faded away;and the intra-articular pressure change would influence the blood supply in femoral head,and could canse transient blood insufficiency.

BACKGROUND:There is lack of an objective and standardized animal model for assessing the therapeutic effect of physical and medication treatment on bone defoct, the effectiveness of operation, as well as the role of bone substitute in the repairing of bone defects.DESIGN:Verified study on the experimental model of bone nonunion in rabbitsSETTING: Department of Orthopaedics in Shenzhen people' s Hospital MATERIALS:Twenty common grade pure New Zealand rabbits of either gender were selected with body mass of (2.5±0.5)kg,aged 6 to 8 months.METHODS :This experiment was carried out at the experimental animal center of Shenzhen people's Hospital between May and August 1999. 1.5cm bone segment (including periosteum)was cut off in the middle of forearm radius in 20 common grade pure NewZealand rabbits,the broken ends were covered with bone wax, 10 weeks later, the bone nonunion status was assessed by macropathological observation, pathohistological and X-ray examination.MAIN OUTCOME MEASURES:Observations on rabbit forearm radius defects by macropathological observation, pathohistological and X-ray examination.RESULTS :Twenty rabbits(40 side radius)were enrolled in this study and weeks later, bone defect region was found filled with fibrous cicatricial tissue without osseous connection ,bone wax was not absorbed, capitulum was ossified with medullary cavity blocked,a small amount of callus formed at both broken ends of fractural bone ,length of bone defect ranged from 0.8 to blocked under optical microscope,chondrocyte and osteocyte could be observed arranging disorderly and covered with fibrous membrane,defect reosseous connection could be detected at defect region at week 10,broken end was ossified and medullary cavity was blocked ,there was small amount of callus appeared at both broken ends displaying irregular shape.CONCLUSION:Bone nonunion experimental animal was successfully established on rabbits in this study, with pathological changes meeting the need of bone nonunion and displaying typical properties,which can be used as reliable and feasible experimental animal model.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.

Objective:To investigate the value of B ultrasonography in the early diagnosis of transient synovitis of hip (TSH) in dogs, and provide the valid base and data for the clinic early diagnosis of TSH. Methods:Eighty 2-3 month old dogs were injected 2‰noradrenalin (NA) into the hip joint induced TSH. We observed 5 assessments that included the 99mTC-MDP triphasic imaging, X ray, B ultrasonography, the synovial lfuid and the pathological tissue check in different time. Results:Early course of TSH presented the synovium of joint hemangiectasis, hyperaemia, synovium villus hyperplasia, edema, and joint inflammatory exudation. The ischemia of local blood supply of the femoral head was observed by 99mTC-MDP triphasic imaging. Ultrasonography showed the broadening of the anterior space of the hip, but the X ray showed no valid changes.Conclusion:B ultrasonography can report the early changes of TSH and may be used in the early diagnosis of TSH in children.

Objective To evaluate surgical indications and method for regional pancreatoduodenectomy combined with blood vessel reconstruction. Methods Forty-four patients underwent pancreatoduodenectomy combined with superior mesenteric vein and portal vein(smv-pv) resection and reconstruction between April 1994 and March 2001.Among them superior mesenteric artery (SMA) and hepatic artery (HA) were reconstructed in 4 and 2 cases, respectively. Partial resection of the anterior wall of the inferior vena cava was performed in one case for tumor invasion. Results The overall mortality was 7.1%,with no complications. The resected endothelium or margins of the blood vessel and pancreas were microscopically tumor free in all cases. Histological specimen examinations demonstrated adenocarcinoma of pancreas head in 43 cases, neuroendocrine adenocarcinoma was diagnosed in one case. Patients were followed-up from 3～87 months with 2 cases lost after PV/SMV for pancreatoduodectomy. Six patients have survived more than 5 yearsand 21 cases more than 3 years. Conclusion Regional pancreatoduodenectomy combined with reconstructionof blood vessel could increase tumor resection rate in properly selected patients and could be performed safely without increased morbidity and mortality.

The incidence of deep vein thrombosis has increased year by year in the world, and it has become one of the serious diseases that threaten human life. In severe cases, it can lead to fatal pulmonary embolism. The pathogenesis of deep vein thrombosis is the result of the interaction of multiple genetic environmental factors. Therefore, regulation of gene expression by modification of DNA and histones may help to further reveal the pathogenesis of deep vein thrombosis, and reactivation or silencing of some genes that are inhibited or overexpressed by aberrant methylation may be a major therapeutic target for deep vein thrombosis. The author reviews the current status of DNA methylation studies related to deep vein thrombosis.

To investigate the dose effect of isocenter difference during IMRT dose verification with the 2D chamber array. The samples collected from 10 patients were respectively designed for IMRT plans, the isocenter of which was independently defined as P(o), P(x) and P(y). P(o) was fixed on the target center and the other points shifted 8cm from the target center in the orientation of x/y. The PTW729 was used for 2D dose verification in the 3 groups which beams of plans were set to 0 degrees. The γ-analysis passing rates for the whole plan and each beam were gotten using the different standards in the 3 groups, The results showed the mean passing rate of γ-analysis was highest in the P(o) group, and the mean passing rate of the whole plan was better than that of each beam. In addition, it became worse with the increase of dose leakage between the leaves in P(y) group. Therefore, the determination of isocenter has a visible effect for IMRT dose verification of the 2D chamber array, The isocenter of the planning design should be close to the geometric center of target.
Gamma Rays ; Humans ; Radiotherapy Dosage ; Radiotherapy, Intensity-Modulated ; instrumentation ; methods

Objective To characterize the cardiac magnetic resonance (CMR) features of peripartum cardiomyopathy(PPCM) and idiopathic dilated cardiomyopathy(IDCM), and to explore the value of MRI in the diagnosis of PPCM. Methods Ten cases of PPCM and 10 cases of Idiopathic dilated cardiomyopathy (IDCM) were included in this study. With 1.5 T MRI scanner, the heart shape (atrioventricular size, hypertrabeculation, thickness of the thinnest ventricular wall), function (ventricular wall movement and the overall function), cardiomyopathy perfusion were comprehensively evaluated. Paired samples t?test and Fisher exact probability method were used for statistical analysis. Results Between PPCM and IDCM group, there was no statistical significant difference in the atrioventricular size, cardiac output(CO), end diastolic volume(EDV), ejection fraction (EF), end systolic volume (ESV) and stroke volume (SV) (P>0.05). IDCM and PPCM group both showed ventricular wall thinning on MRI, with 4 cases of PPCM and 3 cases of IDCM presenting hypertrabeculation in the left ventricular apex. Seven cases of PPCM and 4 cases of IDCM depicted left ventricular local dysfunction, while 3 cases of PPCM and 6 cases of IDCM had abnormal integral movement. Two cases of PPCM appeared local delayed enhancement, while 4 cases of IDCM showed intramural delayed enhancement. After one year of follow?up, heart function recovered in 10 cases of PPCM and 4 cases of IDCM. Conclusions MRI diagnosis using multiple sequences is an ideal method in the evaluation of PPCM. Although there were no differences in cardiac morphology and function between PPCM and IDCM, the prognosis of PPCM is better than IDCM.

Objective: To further explore TCE-induced hepatotoxicity and its mechanisms by identification of trichloroethylene (TCE) induced abnormal histone methylation in human liver cells. Methods: L-02 cells were treated with 0 and 8 mmol/L TCE for 24 h. Histones were extracted by acid. Liquid chromatography electrospray ionization tandem mass spectrometry (ESI-LC-MS/MS) were used to identify and quantify TCE related histone methylations. TCE induced abnormal methylation of H3K79 me2 and H3K79 me3 were validated by Western blot analysis. The further analysis of the function of histone abnormal methylation modifications were done by single cell gel electrophoresis (SCGE) and Western blot analysis of p53 and ɤH2AX. Results: After treatment with TCE for 24 h in L-02 cells, the 36 TCE related histone methylation sites in 28 peptide segments were identified by MS. After treatment with TCE in concentrations of 0 and 8.0 mmol/L in L-02 cells for 24 h, the relative expression level of histone H3K79 me3 were 1.00±0.06, 0.70±0.09 (t=15.01, P=0.015); the relative expression level of histone H3K79 me2 were 1.00±0.05, 0.74±0.07 (t=16.69, P=0.018); the Olive Tail Moment about DNA damage were 1.46±0.28, 3.12± 0.68 (t=15.22, P=0.018); the relative expression levels of p53 were 1.00±0.04, 1.24±0.04 (t=18.71, P= 0.012); and the relative expression levels of ɤH2AX were 1.00 ± 0.03, 1.56 ± 0.11 (t=8.32, P=0 045). Conclusion TCE can induce changes in the relative expression level of H3K79 me2 and H3K79 me3 in L-02 cell, and induce DNA damage, suggesting that TCE may induce changes in the relative expression level of H3K79 me2 and H3K79 me3 by DNA damage.