单侧咀嚼大鼠三叉神经节内PPTAmRNA及其相应蛋白的表达研究

目的:观察磨除单侧后牙造成单侧咀嚼的大鼠三叉神经节内P物质(substance P,SP)及编码 SP 的前速激肽原 A(PPTA)mRNA 的表达情况,进一步探讨颞颌关节病的发病机制.方法:Wistar 雄性大鼠48只,随机分为6组,包括3个实验组及相应对照组,每组8只.实验组动物磨除右侧上、下颌磨牙,人为造成单侧咀嚼.双侧三叉神经节切片行 SP 免疫组化(SABC法)和原位杂交反应.光镜观察拍片,并用 Image Pro Plus 5.1 图像分析软件进行测定.结果与对照组比较,用SPSS10.0软件进行统计分析.结果:每一实验组咀嚼侧和非咀嚼侧三叉神经节内 SP 免疫阳性神经元百分比与各自对照组比较显著降低(p<0.01,p<0.05),非咀嚼侧明显低于咀嚼侧(P<0.01,p<0.05).原位杂交结果显示,每一实验组咀嚼侧与非咀嚼侧三叉神经节中 PPTAmRNA 阳性神经元的数量较各自对照组明显增高(p<0.01),非咀嚼侧明显高于咀嚼侧(p<0.01,p<0.05).结论:三叉神经节内 SP 和 PPTAmRNA 参与了单侧咀嚼引起的颞颌关节病的病理过程,且两侧三叉神经节内SP释放量、PPTAmRNA 合成量不同.     

舌神经切断吻合后菌状乳头味蕾再生的实验研究

目的：了解舌神经损伤后味蕾的变化，以及切断的舌神经用神经外膜吻合修复后舌菌状乳头和味蕾的再生能力。方法：模拟临床将SD大鼠一侧舌神经钳夹或切断损伤，即刻用神经外膜吻合的方法修复切断的舌神经．分别于术后20d和100d时用体视显微镜、组织显微镜、扫描电镜观察鼠舌菌状乳头和味蕾的形态和数目。结果：术后20d时，无论钳夹损伤或切断舌神经，切断的舌神经无论是否用神经吻合修复，舌神经损伤侧的菌状乳头和味蕾均萎缩、退化，数目明显减少；术后100d时，舌神经钳夹损伤和舌神经切断后即刻吻合修复的动物，退化、萎缩的菌状乳头和味蕾已完全再生，形态和数目恢复正常。而舌神经切断后未吻合的动物，萎缩、退化的菌状乳头和味蕾无明显再生恢复。结论：钳夹损伤舌神经，萎缩、退化的菌状乳头和味蕾可以自行再生恢复；切断舌神经，萎缩、退化的菌状乳头和味蕾不能自行再生恢复；用神经外膜吻合修复后，萎缩、退化的菌状乳头和味蕾可以再生恢复。舌菌状乳头和味蕾形态、数目的再生恢复，可以作为舌神经修复成功的客观指标。     

不同缝合法对皮肤圆形缺损瘢痕形成的实验观察

目的:通过动物实验证明荷包缝合结合II期梭形切除/线形缝合法在关闭面部皮肤圆形缺损手术中的可行性和优越性。方法:选取14只成年SD大鼠,将右侧面部设为实验组,左侧为对照组。实验侧面部切除直径为0.5cm的圆形全厚皮片,通过荷包缝合关闭皮肤缺损。术后28d测量瘢痕各项指标后行II期瘢痕梭形切除/线形缝合,14d后再测量最终瘢痕长度。对照侧面部切除短径为0.5cm的梭形皮片,线形缝合14d后测量瘢痕长度。SPSS12.0软件分析数据。结果:实验组I期术后形成椭圆形瘢痕,面积平均仅为原皮肤缺损的18%;结合II期梭形切除/线形缝合,术后瘢痕长度为(0.516±0.052)cm。对照组术后瘢痕长度为(1.341±0.122)cm。SPSS12.0分析两组数据,P<0.01,具有统计学意义。结论:在缝合面部圆形缺损时,荷包缝合结合II期梭形切除/线形缝合相比直接梭形切除/线形缝合具有一定优越性。     

Validity of radiographic evaluations of bone formation in a rat calvaria osteotomy defect model

OBJECTIVE: The objective of this study was to evaluate the validity of radiographic evaluations of bone formation in a critical-size rat calvaria osteotomy defect model. METHODS: Bilateral, critical-size ( [symbol in text] 6 mm) calvaria osteotomy defects in 30 adult Sprague-Dawley rats treated with a rat platelet-rich plasma preparation or control treatments were evaluated by radiographic and histometric measures following a 4- or 8-week healing interval. Standardized radiographic images of the rat calvaria gross specimens were used to assess bone formation within the defect sites by visual evaluation of the grey scale by three masked examiners. The most central portion of each defect site was subject to histometric analysis using a PC-based image analysis system. Kappa statistics and percentage agreement between the radiographic and histometric analysis were estimated. RESULTS: Radiographic evaluations of bone formation are associated with significant weaknesses poorly representing actual healing events; kappa statistics (0.17) denoting slight agreement beyond chance. Perfect agreement between the histologic and radiographic analysis for defect sites showing complete and partial histologic bone fill was achieved 63% and 50% of the time, respectively. Agreement reached only 20% for sites with no/limited bone fill. When no/limited and partial bone fill occurred, the radiographic analysis tended to overestimate bone fill and underestimate bone fill when complete closure of the defect sites was observed in the histologic analysis. CONCLUSION: Low accuracy was observed when radiographic evaluations were employed in identifying and characterizing bone fill in the rat calvaria osteotomy defects. Assessment of bone healing in animal models aiming at treatment recommendations for clinical application must not solely be based on radiographic analysis, but should be confirmed using histologic observations.     

Bone resorption and serum levels of vitamin D metabolites in the hypophosphataemic rat

The supplementation of a low phosphate diet with vitamin D has been shown to result in an increase in bone resorption in the hypophosphataemic rat. The aim of the present study was to determine if administration of vitamin D to rats fed a vitamin D and phosphate-depleted diet would result in an increase in the circulatory levels of the active vitamin D metabolite 1,25(OH)₂D₃ and an associated increase in bone resorption. Three groups of weanling Sprague-Dawley rats were used. The first group consisted of control animals on a normal laboratory stock diet and the second and third groups were experimental animals receiving a vitamin D-and phosphate-deficient diet with the third group receiving vitamin D supplementation. All animals were housed in the dark. After 30 days on the diet the experimental animals received 0.1 mmol NaH₂PO₄ by intraperitoneal injection. Blood was sampled at zero, 3, 6, 18 and 48 h post-injection and analysed for the vitamin D metabolites 25(OH)D₃ and 1,25(OH)₂D₃, calcium and inorganic phosphate (Pi). The serum analyses revealed that the level of 25(OH)D₃ in the hypophosphataemic animals was significantly lower than that of the control animals. However the 1,25(OH)₃D₃ level was initially significantly higher, then dropped to the control level at 18 h post-intraperitoneal injection of phosphate. Further, the serum levels of 25(OH)D₃ and 1,25(OH)₂D₃, calcium and Pi in the hypophosphataemic animals supplemented with vitamin D were significantly higher than those of the vitamin D-deficient animals. Also the vitamin D-supplemented animals exhibited significantly greater levels of bone resorption. These results therefore, are consistent with a role of 1,25(OH)₂D₃ in bone resorption in hypophosphataemic rats.     

盐酸丁咯地尔对SD大鼠坐骨神经损害后神经元的保护作用

目的:观察盐酸丁咯地尔对坐骨神经损害后神经元病理形态的改变.方法:以SD大鼠坐骨神经建立周围神经损害动物模型,观察背根神经节病理切片(光镜、电镜)中的神经元变化情况,考察盐酸丁咯地尔对坐骨神经损害后神经元的影响.结果:盐酸丁咯地尔给药4周后能显著地减轻背根神经节内神经元细胞坏死.结论:盐酸丁咯地尔对坐骨神经损害后神经元有确切的保护作用.     

玉屏苍耳散对实验性AR大鼠维甲酸相关孤儿核受体γt及相关因子表达的影响

目的探究玉屏苍耳散对实验性变应性鼻炎(AR)大鼠维甲酸相关孤儿核受体γt(RORγt)及相关因子表达的影响。方法将40只SD大鼠随机分为正常组,模型组,玉屏苍耳散组,西替利嗪组,每组各10只。采用卵清蛋白致敏法构建AR大鼠模型。灌胃给药,给药剂量按照大鼠体重计算:玉屏苍耳散组(10 g/kg),西替利嗪(2 mg/kg),正常组以及模型组灌胃给予等量的生理盐,酶联免疫法检测各组大鼠鼻粘膜组织细胞因子IL-17、TNF-α、IL-23以及IL-6的分泌水平;HE染色检测各组大鼠鼻黏膜组织损伤程度;采用免疫组化法检测各组大鼠鼻黏膜组织中RORγt蛋白的表达;通过RT-PCR检测各组大鼠鼻黏膜组织中RORγt、IL-17以及IL-23 m RNA的表达。结果与正常组比较,模型组大鼠鼻部症状评分、鼻粘膜厚度、细胞因子IL-17、TNF-α、IL-23、IL-6表达及RORγt、IL-17以及IL-23 m RNA表达明显升高(P<0.05);与模型组相比,玉屏苍耳散组与西替利嗪组上述指标均明显降低(P<0.05)。结论玉屏苍耳散可调控RORγt以及相关因子IL-17、IL-23的表达,发挥对AR具有免疫调节作用。     

依托咪酯对新生大鼠认知功能的影响及其机制研究

目的 探讨新生大鼠依托咪酯麻醉后近远期认知功能及烟碱型乙酰胆碱受体（nAChR）功能的变化。方法 将7 d龄新生大鼠随机分对照组、依托咪酯组、激动剂组,分别单次腹腔注射生理盐水10 ml/kg、依托咪酯5 ml/kg、依托咪酯5 ml/kg+α7nAChR激动剂PNU-282987 5 ml/kg,每组16只。麻醉清醒后2 h和4周,各组随机取8只大鼠进行行为学测试,完成测试后立即取双侧海马组织,采用qRT-PCR和Western blotting检测α7nAChR、乙酰胆碱酯酶（AChE） mRNA和蛋白。结果 麻醉清醒后2 h,依托咪酯组洞板实验探洞次数少于对照组和激动剂组（P <0.05）;3组Y电迷宫实验中正确反应次数和全天总反应时间比较,差异无统计学意义（P >0.05）。麻醉清醒后4周,3组洞板实验探洞次数、Y电迷宫实验中正确反应次数和全天总反应时间比较,差异无统计学意义（P >0.05）。麻醉清醒后2 h,依托咪酯组海马组织α7nAChR mRNA相对表达量低于对照组和激动剂组（P <0.05）,AChE mRNA和蛋白相对表达量高于对照组和激动剂组（P <0.05）。麻醉清醒后4周,3组α7nAChR、AChE mRNA和蛋白相对表达量比较,差异无统计学意义（P >0.05）。结论 新生大鼠依托咪酯麻醉对近期学习记忆能力和远期认知能力无明显影响,但可一过性影响新生大鼠近期探索能力,其机制可能与抑制海马nAChR功能有关。     

新生大鼠单侧基底核出血对神经功能的影响观察

目的：探析新生大鼠单侧基底核出血对神经功能产生的影响。方法：抽取出生第10 天的54 只新生大鼠,根据随机数字分组的原则随机分为正常对照组（normal control group,NC,n=18）、假手术组（sham operation group,SH,n=18）、脑出血组（cerebral hemorrhage group,CH,n=18）,通过立体定位仪将25 μL 自体血注入大鼠左侧脑尾状核位置,完成脑基底核出血模型的建立工作。NC 组不采取任何处理干预,给予正常饲养,SH 组在大鼠左侧尾状核位置刺入微量注射泵针头,不注血,其他处理和CH 组相同。对比3 组大鼠不同时间点神经功能评分、白天开场实验结果、晚上开场实验结果等指标。结果：大鼠在造模之后神经功能呈现不同程度损伤,SH 组大鼠有着相对轻微的神经受损症状,能在短时间内恢复,在手术后72 h 能全部康复;CH 组大鼠发生肌张力改变以及偏瘫的情况;与SH 组、NC 组相比,CH 组大鼠在d 1、d 3 的神经功能评分明显较高（P<0.05）;SH 组、NC 组大鼠在d 1、d 3、d 7、d 14 的神经功能评分对比差异不大（P>0.05）;和SH 组、NC 组大鼠白天开场实验结果相比,CH 组大鼠理毛次数、直立次数、穿越格子数等明显较多（P<0.05）;和SH 组、NC 组大鼠晚上开场实验结果相比,CH 组大鼠直立次数、穿越格子数等明显较多（P<0.05）。结论：大鼠在发育期会出现单侧基底核局灶性出血损伤症状,有着可代偿性、可修复性等基本特征,大部分不会出现严重神经运动功能障碍遗留的情况,但会引起非选择性注意水平降低、自主活动增多等症状。