1.Construction and evaluation of intracerebral capsular hemorrhage models in rats
Rufei DAI ; Jun CAI ; Ning LIU ; Fengyi ZHU ; Xiaoming LI
Chinese Journal of Tissue Engineering Research 2006;10(42):211-213
BACKGROUND: A stable and exact animal model is the necessary tool and basis for studying hemorrhagic cerebrovascular diseases.OBJECTIVE: To establish and evaluate the intracerebral capsular hemorrhage models in rats.DESIGN: A randomized and controlled animal experiment.SETTING: Second Hospital Affiliated to Xuzhou Medical College; First Hospital Affiliated to Nanjing Medical University.MATERIALS: This experiment was carried out in the animal experimental center of Nanjing Medical University during May to November 2002.Totally 35 SD rats were randomized into two groups: experimental group (n=30) and sham-operation group (n=5).METHODS: ① Autoblood was injected into the intracerebral capsule of rats to create intracerebral capsule hemorrhage models with stereotaxy in the experiment group. ②Scoring was conducted according to 5-point neurological scoring criteria from ZeaLonga, somatic sensation and motor function of rats were observed. ③Somatosensory evoked potential(SEP) of rats was detected pre- and post-operation under anesthetic state. ④ After determination of SEP, the rats were sacrificed under anesthetic state. Brains were taken out to made slices, then sections were stained with haematoxylin and eosin. Changes in haematoma and histomorphology were observed at the largest focus under optical microscope.MAIN OUTCOME MEASURES: ①Nerve function scoring; ②Latent period of various waves of SEP; ③ Observation of brain tissue morphology.RESULTS: Totally 35 rats entered the stage of result analysis. ①Appearance of obvious paralysis of the rats suggested the modeling was successful. The successful rate of this experiment was 93.3%(28/30). Significant difference existed in neurological scoring between experimental group [(2.74±0.46)points] and sham-operation group (0 point)(P<0.05). ②SEP showed that the latent periods of various waves of experimental group after operation were significantly delayed than those before operation and those of sham-operation group [P1: (15.72±0.78) ms, (10.69±0.52) ms, (10.73±0.48) ms;Nl: (17.95±1.27) ms,(13.21±1.31) ms, (13.34±1.27) ms;N2:(21.16±1.62) ms, (15.42±1.46) ms,(15.58±1.44) ms;N3:(24.86±1.58) ms, (18.72±1.76) ms, (18.99±1.67) ms,P<0.05]. ③In the shamoperation group, a few red blood cells were scattered in the peripheral area of needle channel were found, but hemorrhagic focus was not; In the experimental group, irregular or oval blood clots presented in the left internal capsule area. In about a low-fold visual field, brain tissue in the surrounding of hemorrhagic focus was loosened and swelled, and pathological changes were obviously severer than those in the sham-operation group.CONCLUSION: Intracerebral capsular hemorrhage induced by injection of autoblood with stereotaxy is more close to clinical situation, and it is easy to operate and has good reproducibility.
2.The effects of NOD2 gene on the proliferation and apoptosis of human tongue squamous cell carcinoma Tca8113cells
Mingyang SU ; Hongyu YANG ; Rufei ZHU ; Huijun YANG ; Shiyue SHEN ; Lamei WU ; Feng WANG
Journal of Practical Stomatology 2014;(4):510-514
Objective:To explore the effects of NOD2 gene on the proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells.Methods:NOD2 expression vector(NOD2-pEZ-M29)and NOD2-shRNA vector were established,then were trans-fected into Tca8113 cells respectively.Expressions of HBD-2 and NOD2 in the cells was detected by RT-PCR and Western blot.Cell proliferation was examined by MTT assay and apoptosis by flow cytometry at 48h post transfection.Results:Compared with the control group,the expression of NOD2 and HBD-2 in NOD2-pEZ-M29 transfection group was significantly higher and markedly lower in NOD2-shRNA group.The proliferation rate of Tca8113 cells was markedly lower in NOD2-pEZ-M29 transfection group and signifi-cantly higher in NOD2-shRNA group while the apoptosis rate was significantly higher in NOD2-pEZ-M29 transfection group and sig-nificantly lower in NOD2-shRNA group.Conclusion:In Tca8113 cells NOD2 expression was positively correlated with HBD-2 ex-pression.NOD2 gene may promote the apoptosis,inhibit the proliferation of Tca8113 cells.