Tumor invasion model is an important tool to study the mechanisms of tumor mvasion and to evaluate prevention and treatment of cancer.Compared to the in vitro model,in vivo model can simulate the tumor microenvironment and tumor mvasion biological behaviour better.Morphological observation and invasion-related protein detection can be used to evaluate the animal invasion model,which are widely used in basic and clinical research now.

Objective To investigate the protective role of carbon monoxide releasing molecule-2 (CORM-2) in post-resuscitation myocardial dysfunction (PRMD) in rat models of cardiopulmonary resuscitation (CPR).Methods Cardiopulmonary resuscitation model was established after cardiac arrest induced by ventricular fibrillation.Male healthy Sprague-Dawley (SD) rats were randomly (random number) divided into 4 groups according to random number table:control group,CORM-2 group,inactive CORM-2 (iCORM-2) group and Sham group,in which the equal volume (1 mL) of 0.2％ DMSO,50 μmol/kg CORM-2,50 μmol/kg iCORM-2 and 0.2％ DMSO were respectively administered into the rats of these groups after resuscitation.The ejection fraction (EF) of left ventricle and myocardial performance index (MPI) were measured to detect the myocardial function by echocardiography at 12 hours after resuscitation.Mitochondrial respiration was assessed with Clark oxygen electrode at the same time.Western blot was used to determine the ratio of mitochondrial cytochrome c (cyt c) to cytoplasmic cyt c as well as caspase-3 level.Multiple comparisons were made by analysis of variance.Results Compared with the control group,higher EF and MPI,higher state Ⅲ respiration rate and respiratory control rate (RCR) of mitochondria,and decreased ratio of mitochondrial cytc/cytoplasmic cyt c and lower caspase-3 level were observed in the CORM-2 group (P ＜ 0.05).However,there were no significant differences in above biomarkers found between iCORM-2 group and control group (P ＞ 0.05).Conclusions The CO released from CORM-2 might improve mitochondrial respiration and PRMD by inhibition of myocardial apoptosis via a mitochondrial pathway.

Objective To investigate the effects of mitochondrial division inhibitor 1 (mdivi-1) in rats after cardiopulmonary resuscitation (CPR) and its mechanism.Methods Fifty Sprague-Dawley (SD) rats were randomly (random number table) divided into sham group (n =8),cardiac arrest (CA) model group (n =14),dimethyl sulfoxide post-treatment control group (DMSO group,n =14),and mdivi-1 post-treatment group (mdivi-1 group,n =14).Asphyxial CA was reproduced in animals,and they were resuscitated by CPR.In the mdivi-1 group or DMSO group,the animals were given mdivi-1 (1.2 mg/kg) or DMSO (0.1％) intravenously after restoration of spontaneous circulation (ROSC).The neurological functions were assessed using neurological deficit score (NDS) determined at 24,48 and 72 hours after CPR.The brain tissues were harvested at 72 hours after CPR.The histopathologic changes were assessed by hematoxylin and eosin (HE) staining,and the normal neuron was counted.The neuronal apoptosis was assessed with terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining,and the expressions of cytochrome C (Cyt-C) protein in mitochondria and cytoplasm from hippocampus were determined by Western Blot.Results NDS in all experiment groups was gradually increased after CPR,and they were significantly lower than thoseo.f the sham group at 24,48,and 72 hours (51.5±3.7 vs.80.0±0.0,59.3±3.6 vs.80.0±0.0,66.7±2.6 vs.80.0±0.0,all P ＜ 0.05).The number of normal pyramidal neurons in the hippocampal CA1 region was markedly reduced (cells/HP:4.4± 1.1 vs.23.1 ± 4.0,P ＜ 0,05),the apoptotic index was significantly increased [(86.9 ± 6.9)％ vs.(3.4 ± 0.8)％,P ＜ 0.05],the expressions of Cyt-C in mitochondria were significantly decreased (A value:0.46±0.18 vs.1.00±0.00,P ＜ 0.05),and the expressions of Cyt-C in cytoplasm were significantly up-regulated (A value:6.65±0.21 vs.1.00±0.00,P ＜ 0.05).Compared with the CA group,NDS at 24 hours and 48 hours in mdivi-1 group was slightly increased (55.2 ± 3.3 vs.51.5 ± 3.7,64.7 ± 2.4 vs.59.3 ± 3.6,both P ＞ 0.05),and it was significantly increased at 72 hours (74.5±2.3 vs.66.7 ± 2.6,P ＜ 0.05),the number of normal pyramidal neurons in the hippocampal CA1 region was markedly increased (cells/HP:16.2±2.4 vs.4.4± 1.1,P ＜ 0.05),the apoptotic index was dramatically reduced [(42.3 ± 3.9)％ vs.(86.9 ± 6.9)％,P ＜ 0.05],the expressions of Cyt-C in mitochondria were significantly increased (A value:0.83 ± 0.22 vs.0.46 ± 0.18,P ＜ 0.05),and the expressions of Cyt-C in cytoplasm were significantly decreased (A value:3.84±0.47 vs.6.65±0.21,P ＜ 0.05).There was no statistically significant difference in above indexes between CA group and DMSO group.Conclusion By inhibiting mitochondrial Cyt-C apoptotic pathway to reduce neuronal apoptosis in rats after CA-CPR,mdivi-1 can improve brain function after CPR.

Objective: Numerous studies have shown that bone marrow-derived mesenchymal stem cells (MSCs) enhance neurological recovery after cerebral ischemia. However, the mechanisms are still not clear. The present study aimed to investigate the beneficial effects of MSCs on global cerebral ischemia induced by cardiac arrest (CA) and the underlying mechanisms. Methods: Rats subjected to asphyxial CA were injected intravenously with MSCs (5×106 ) at 2 hours after resuscitation. Whole brain histopathologic damage scores (HDS) were assessed by histopathology at 3 and 7 days after resuscitation. The distribution of donor MSCs in the brain was evaluated. The expression of tumor necrosis factor-α-induced protein 6 (TSG-6) and pro-inflammatory cytokines in cerebral cortex was assayed. After intravenous infusion of TSG-6 siRNA-MSCs, HDS and pro-inflammatory cytokines were reevaluated at 7 days after resuscitation. Results: Intravenously administered MSCs significantly reduced whole brain HDS after global cerebral ischemia. Immunofluorescence microscopy revealed that donor MSCs were primarily found in cerebral cortex and expressed TSG-6. MSCs treatment significantly increased the expression of TSG-6 and reduced the expression of pro-inflammatory cytokines in cerebral cortex. In addition, intravenous infusion of TSG-6 siRNA-MSCs failed to attenuate brain inflammation. Conclusion: Systemically administered MSCs reduced inflammatory damage to brain in rats with global cerebral ischemia via secretion of TSG-6.
Heart Arrest ; Mesenchymal Stromal Cells

Objective To investigate the effects of bone marrow mesenchymal stem cells (MSCs)treatment on TSG-6 in a rat model of cardiopulmonary resuscitation (CPR).Methods Sprague Dawley (SD) rats were randomly (random number) divided into sham group,phosphate buffer solution (PBS)-treated group and MSCs-treated group.Animals were subjected to asphyxial cardiac arrest followed by CPR.In PBS-treated group or MSCs-treated group,animals were injected intravenously with PBS or MSCs at 2h after resuscitation.Neurological deficit scores (NDS) were assessed at 1,3 and 7 d after CPR.Serum S-100B was assayed using enzyme linked immunosorbent assay (ELISA).Immunofluorescence was performed to detect donor MSCs and the expression of TSG-6 in brain.TSG-6 and proinflammatory cytokines in brain were assayed using real time reverse transcription-polymerase chain reaction (RT-PCR).Western blot analysis was performed to measure the levels of neutrophil elastase (NE) in brain.Multiple comparisons were made by analysis of variance.Results At 3d and 7d,MSCs-treated group demonstrated higher NDS than PBS-treated group (P ＜ 0.01),and serum S-100B levels significantly reduced in MSCs-treated group compared with PBS-treated group (P ＜ 0.01).DAPI-labeled MSCs migrated into the ischemic brain and some DAPI + cells colocalized with TSG-6.Compared with PBS-treated group,MSCs treatment significantly up-regulated the expression of TSG-6 and reduced the expression of NE and proinflammatory cytokines in brain at 3 d and 7 d after CPR (P ＜ 0.05).Conclusion Systemically administered MSCs suppressed inflammatory responses in brain after CPR and improved neurological function in rats possibly via induction of TSG-6.

Objective: Asphyxia and ventricular fibrillation are the two most prevalent causes of cardiac arrest. The study investigated the differences in brain damage after cardiac arrest between asphyxial and ventricular fibrillation cardiac arrests in rats. Methods: Male healthy Sprague-Dawley rats were randomly assigned to the asphyxial group (cardiac arrest of 6 min, n=15), ventricular fibrillation group (cardiac arrest of 6 min, n=15) and sham group (n=5). Neurologic deficit scores and tape removal test were evaluated at 1, 3 and 7 days after cardiopulmonary resuscitation from three groups. Serum S-100B and brain histopathologic damage scores were also examined. Results: There were no differences in neurologic performance at 1, 3 and 7 days after cardiopulmonary resuscitation between the asphyxial group and ventricular fibrillation group (P>0.05, respectively). Serum S-100B level was higher in the asphyxial group at 1, 3 and 7 days, compared with the ventricular fibrillation group (P<0.05, respectively). There were significantly higher histopathologic damage scores at 1, 3 and 7 days in the asphyxial group compared with the ventricular fibrillation group (P<0.05, respectively). Conclusion: Asphyxial cardiac arrest has worse morphologic brain damage compared with ventricular fibrillation cardiac arrest, but the functional brain damage caused by asphyxial cardiac arrest is similar to that caused by ventricular fibrillation cardiac arrest.

OBJECTIVETo study the clinic presentation, radiology, pathology of recurrent ameloblastoma (RAB).

METHODSAll RAB cases accepted treatment in School of Stomatology of Nanjing University during 1996.1-2008.3 were retrospected. The clinic presentation, the radiological classification and pathological classification were summarized and analyzed.

RESULTSThere were 23 RAB patients during 1996.1-2008.3. 3 patients recurred twice, 20 patients recurred once, and 26 cases were found together. 19 patients were performed conservative surgery while 4 patients were performed radical surgery in the primary treatment; 4 patients were performed conservative surgery and 19 patients were performed radical surgery in the recurrence. The radiological classification included 12 cases multicystic, 6 cases unicystic, and 8 cases extraosseous. The pathological classification included 21 cases follicular ameloblastoma, and 5 cases plexiform ameloblastoma. The multicystic after conservative surgery (11 cases) and extraosseous ameloblastoma after radical treatment (8 cases) were more than others according the radiological classification and treatment.

CONCLUSIONConservative surgery has apparent higher recurrences rate than radical surgery. The follicular ameloblastoma has more aggressive biological behave and is more liable to recurred.

Objective To investigate the mechanism of Guiqiyiyuan Ointment (GO) for preventing and treating 12C6+ beam radiation induced lung and kidney bystander effect to provide a new strategy for prevention and treatment of clinical radiation injury. Methods Sixty healthy male Wistar rats were randomly and equally divided into 3 groups: NC group, SR group (simple radiation 2ml/kg), GO group(GO 2ml/kg intragastric administration for 7 days). The right side of the lung was modeled by 12C6+beam radiation. After modeling, the rats were killed at 48h. The left lung, left and right kidney tissues were taken from the rats. The DNA methylation rate was detected by ELISA assay, pathological changes were observed by HE staining, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were detected by immunohistochemistry. Results Compared with the NC group, the level of DNA methylation was decreased significantly (P<0.01), the left lung showed inflammation, no abnormal finding was seen in the left and right kidneys, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were significantly increased in the SR group (P<0.01). Compared with the SR group, the level of DNA methylation was increased significantly (P<0.01), the left lung inflammation became better, and the expressions of Dnmt1, Dnmt3a and Dnmt3b were significantly decreased in the GO group (P<0.01). Dnmt1, Dnmt3a and Dnmt3b proteins were expressed in the cytoplasms of bronchial and renal tubular epithelial cells in all the groups. The NC group presented as light brown-brown staining, showing a weak positive expression, the SR group as brown-brown staining, showing astrong positive expression, and the GO group as light brown-brown staining, showing a moderate positive expression. Conclusion The GO can reduce the bystander effect caused by 12C6+ beam radiation, and its mechanism is related to improving the level of DNA methylation.

Objective To observe the protection and distribution of bone marrow mesenchymal stem cells (MSCs) by distinct intravenous infusion time on renal ischemia reperfusion injury (IRI) in rats.Methods We used unilateral nephrectomy and contralateral vascular occlusion method to establish renal IRI model in rats.The experimental groups which received 2 × 106 MSCs infusion through the tail vein,were subsequently divided into 3 subgroups:2 h pre-reperfusion (PreOp,n =16),immediately after reperfusion (Op,n =16),6 h post-reperfusion (PostOp,n - 16).The control groups included sham operation group (n =16) and ischemia group (n =16).Chemotaxis of DAPI-labeled MSCs was detected 6 h after administration in the IR kidney.Renal function was detected at 6,24,and 48 h respectively after operation. Forty eight h after operation,the renal tissues were harvested to observe the pathological changes by HE staining and the tubular epithelial cell apoptosis via TUNEL assay.Results MSCs were found in the experimental groups after IR in the kidney,most in PostOp group.Twenty-four and 48 h after reperfusion,there was no significant difference in Cr and BUN between the experimental groups and sham operation group (P＞0.05),but the levels of Cr and BUN in the experimental groups were significantly lower than in the IR group (P＜ 0.05). As compared with IR group,the renal pathological injury was alleviated,the number of apoptotic cells was decreased in the experimental group,most significantly in PostOp group (P＜0.05).Conclusion MSCs can reduce the inflammatory response and inhibit renal tubular cell apoptosis in rat renal IRI.Post-reperfusion administration of MSCs leads to the best chemotaxis efficiency and protection.

Objective:To explore the effect of the pharyngeal pressure feedback training on pharyngeal constriction in persons with swallowing disorders caused by brainstem lesions.Methods:Twenty patients with disordered swallowing caused by a brainstem lesion were randomly divided into a control group and an intervention group, each of 10. Both groups received routine swallowing training including oral sensorimotor training, neuromuscular stimulation and balloon catheter dilation, while the intervention group was additionally provided with two weeks of pharyngeal pressure feedback training. Before and after the treatment, both groups were evaluated in terms of the peak pressure of superior and inferior pharyngeal constriction, endoscopically and also using the functional oral intake scale.Results:There was no significant difference between the two groups in any measure before the intervention. Afterward, both groups had improved significantly by all of the measurements, but the average peak upper pharyngeal pressure, the average intake scale score and the endoscopy results of the intervention group were all significantly better than the control group′s averages.Conclusions:Pharyngeal pressure feedback training can significantly relieve swallowing disorders caused by brainstem lesions.