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背景:氟桂嗪和转化生长因子β(TGF-β)都具有抗脑缺血损伤作用,但两者之间是否存在某种联系,目前还不明确。目的:通过研究氟桂嗪对沙鼠脑缺血再灌注后脑内转化生长因子I型,II型受体(TβRI,II)基因表达的影响,探讨氟桂嗪与TGFβ信号转导途径在抗脑缺血损伤方面的联系。设计:随机对照的实验研究。地点和对象:实验在中南大学湘雅二医院中心实验室完成。健康雄性蒙古沙鼠60只,9月龄,体质量(90±5)g,随机分为脑缺血组、氟桂嗪治疗组、假手术组、正常对照组,其中脑缺血组、氟桂嗪治疗组各有缺血再灌6h,1,3,7d组,共计10组,每组6只。干预:夹闭双侧颈总动脉法制作沙鼠脑缺血再灌注模型,氟桂嗪治疗组实验前1d给沙鼠喂食氟桂嗪犤按20mg/(kg·d)犦。采用原位杂交检测TβRI,II基因表达情况,用苏木精-伊红染色方法观察脑组织病理变化。主要观察指标:脑组织病理变化,及脑内TβRI,IImRNAs的表达。结果:氟桂嗪治疗组在再灌注各个时间点脑组织损伤程度均明显轻于脑缺血组。各组沙鼠脑组织的神经元和胶质细胞胞浆均有TβRI,IImRNAs阳性表达。棕褐色颗粒主要位于神经元和胶质细胞胞浆中,但表达程度有所不同。假手术组的TβRI,IImRNAs的表达较正常对照组稍高但无明显差异(P>0.05)。氟桂嗪治疗组中缺血再灌注6h,1d,3dTβRI,IIm 相似文献
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目的:评价一期后、前路手术治疗发育性颈椎管狭窄合并巨大椎间盘突出的特点及疗效。方法:回顾分析2002年2月至2006年1月接受该术式的21例患者的临床资料,对手术前后临床表现、JOA评分、MRI影像变化以及术后神经功能改善率及并发症进行总结分析。结果:术后未发生神经系统症状加重、感染及内固定失效等并发症。获得1年以上随访者21例,平均随访时间为29.2个月,术后3个月、12个月时神经功能改善率分别为:58%、66%;16例获得2年以上随访者神经功能改善率为71%。结论:一期后、前路手术治疗发育性颈椎管狭窄合并巨大椎间盘突出可缩短治疗周期,相对节省治疗费用,同时解除了硬膜囊前、后方的压迫,减压彻底,疗效满意。 相似文献
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目的总结一期前路病灶清除植骨内固定治疗脊柱结核的临床效果。方法对13例平均年龄42·3岁,平均病程5·6个月的脊柱结核患者,采用一期病灶清除结合植骨内固定,手术前后配合正规化疗,根据X线片观察脊柱融合时间,手术前后后凸角度变化以及按照Frankel分级的神经功能变化。结果全部病例伤口均一期愈合,未出现严重并发症。平均随访时间17·3个月,植骨界面骨性融合时间平均5·6个月。后凸平均矫正度数为11·7°,7例术前伴有神经损害症状者Frankel分级平均提高1·1级。结论一期病灶清除植骨内固定治疗脊柱结核,骨结构重建可靠,治疗过程相对简化,住院周期缩短,效果肯定。 相似文献
5.
慢性湿疹是一种比较常见的慢性、顽固性皮肤病,临床主要表现为多型皮疹,分布对称,易反复发作,慢性经过,瘙痒剧烈等特征,病因复杂,预防和治疗困难。1999年1月-2003年1月,我院应用静脉封闭疗法治疗慢性湿疹50例,疗效满意,现报告如下。 相似文献
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目的:分别采用高效液相色谱法和气相色谱质谱联用法测定白念珠菌麦角甾醇含量,并对这两种方法进行比较。方法:采用浓度梯度递增法诱导白念珠菌耐药株,采用KONT真菌显色MIC药敏系统鉴定耐药性。构建6、12、24h生物被膜,采用高效液相色谱法和气相色谱质谱联用法测定麦角甾醇含量。结果:高效液相色谱法检测出麦角甾醇含量,最低检出浓度为0.05mg/L,而气相色谱质谱联用法未检测出麦角甾醇含量。结论:高效液相色谱法能准确测定白念珠菌麦角甾醇含量。与气相色谱质谱联用法比较,高效液相色谱法简单、高效和可靠。 相似文献
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目的 探讨联合检测血清肿瘤标志物对肝癌诊断的价值.方法 选择55例原发性肝癌( PHC)患者、120例各型肝炎患者和30例正常健康体检者,分别检测血清甲胎蛋白(AFP)、甲胎蛋白异质体(AFP-L3)、糖类抗原( CA199)、癌胚抗原(CEA)、血清铁蛋白(SF)、a-L-岩藻糖苷酶(AFU)、γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)检测,用四格表法计算灵敏度(Se)、特异度(Sp)、Youden指数、预测值(PV)、似然比(LR)、比数积(OP)、准确度(AC)等特征参数.结果 AFP-L3的受试者工作特性曲线下面积最大,其Sp、Youden指数、OP、LR+、PV+、AC也最大,Se、PV-最大为GGT.平行试验中AFP/AFP-L3/CEA组合Se为94.6%,PV-为97.2%,Youden指数为64.6%,AC为76.6%;系列试验中AFP+ AFP-L3+ GGT组合Sp为97.3%,LR+为25.2,Youden指数为64.6%,AC为89.3%.结论 PHC筛查检测应首选AFP/AFP-L3/CEA平行试验组合,确诊检测应首选AFP+AFP-L3+GGT系列试验组合. 相似文献
8.
42只家兔分为对照组与3个实验组,通过膈神经放电积分信号判断动物呼吸频率、幅度的变化,初步分析了由谷氨酸钠引起延髓最后区细胞兴奋而产生的呼吸效应的递质基础。实验组在延髓最后区微量注射谷氨酸钠前经脑室分别注入妥拉苏林、肉桂硫胺、阿托品,对照组则注入与阻断剂等量的生理盐水。结果表明,脑室预先注入妥拉苏林、肉桂硫胺后,最后区微量注射谷氨酸钠的呼吸增频反应明显被抑制,与对照组比较,反应差别有显著的统计学意义;脑室注射阿托品后,最后区微量注射谷氨酸钠,动物出现与对照组类似的浅快呼吸,但呼吸反应达高峰的时间推迟。本工作提示:最后区细胞兴奋后可能通过去甲肾上腺素、5-羟色胺等物质的作用引起呼吸频率加快反应,乙酰胆硷则在最后区注射谷氨酸钠引起的呼吸兴奋效应中可能起易化作用。 相似文献
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BACKGROUND: Overexpression of α-synuclein can induce cell apoptosis. RNA interference (RNAi) may block specific gene function and cause gene silencing. OBJECTIVE: To construct a specific and effective RNAi plasmid for the α-synuclein gene and investigate if RNAi can block apoptosis in HEK293 cells, induced by overexpression of wild-type α-synuclein.
DESIGN, TIME AND SETTING: A contrast experiment based on genetically engineered cytobiology was performed at the State Key Lab of Medical Genetics of China, Xiangya Medical College of Central South University, between October 2004 and October 2008.
MATERIALS: HEK293 cells and pBSHH1 plasmid were provided by the State Key Lab of Medical Genetics of China; OligDNA sequence by Sagon Bioengineering Company, Shanghai; Lipofectamine 2000 by Invitrogen, USA; α-synuclein monoclonal antibody, Hoechst 33258, and MTT by Sigma, USA; Horseradish peroxidase-coupled goat anti-rat IgG by KPL, USA; FACSan flow cytometry by BD, USA.
METHODS: Four target sites were used to construct hairpin RNA pBSHH1 vectors - pSYNi-1, pSYNi-2, pSYNi-3 and pSYNi-4 - which were cloned in the pBSHH1 plasmid. HEK293 cells were transfected using Lipofectamine 2000. In addition, a non-transfect group and a negative plasmid transfect group were established. The cultured HEK293 cells were processed as follows: transfection of blank plasmid (blank control group), transfection of α-synuclein-pEGFP and RNAi negative vector (negative control group), and transfection of α-synuclein-pEGFP and pSYNi-1 (transfection group). Cells in all groups were transfected with Lipofectamine 2000 for 48 hours.
MAIN OUTCOME MEASURES: Expression of α-synuclein mRNA and protein were detected by RT-PCR and Western blot. Cell morphology was observed under an inverted fluorescence microscope; cell viability was measured using MTT method; and cell apoptosis was determined with Annexin V-PE flow cytometry.
RESULTS: α-synuclein mRNA and protein expressions were significantly decreased in the pSYNi-1 group when compared with the non-transfect and negative plasmid transfect groups (P 〈 0.05). The expressions were partially decreased in the pSYNi-2 group, but there was no significant difference in the pSYNi-3 and pSYNi-4 groups. Hoechst staining indicated that cell nuclei were enlarged in the negative control group, coloring was not uniform, and chromatin was accumulated and appeared spot-like. The nucleus coloring was uniform in the transfection group compared to negative control group. Cell viability in the negative control group was significantly lower than blank control group with cell apoptosis being significantly increased (P 〈 0.05). In comparison with negative control group, cell viability was significantly increased in the transfection group and cell apoptosis was significantly decreased (P 〈 0.05).
CONCLUSION: pSYNi-1 can inhibit α-synuclein gene expression and block apoptosis of HEK293 cells induced by overexpression of wild-type α-synuclein. 相似文献
DESIGN, TIME AND SETTING: A contrast experiment based on genetically engineered cytobiology was performed at the State Key Lab of Medical Genetics of China, Xiangya Medical College of Central South University, between October 2004 and October 2008.
MATERIALS: HEK293 cells and pBSHH1 plasmid were provided by the State Key Lab of Medical Genetics of China; OligDNA sequence by Sagon Bioengineering Company, Shanghai; Lipofectamine 2000 by Invitrogen, USA; α-synuclein monoclonal antibody, Hoechst 33258, and MTT by Sigma, USA; Horseradish peroxidase-coupled goat anti-rat IgG by KPL, USA; FACSan flow cytometry by BD, USA.
METHODS: Four target sites were used to construct hairpin RNA pBSHH1 vectors - pSYNi-1, pSYNi-2, pSYNi-3 and pSYNi-4 - which were cloned in the pBSHH1 plasmid. HEK293 cells were transfected using Lipofectamine 2000. In addition, a non-transfect group and a negative plasmid transfect group were established. The cultured HEK293 cells were processed as follows: transfection of blank plasmid (blank control group), transfection of α-synuclein-pEGFP and RNAi negative vector (negative control group), and transfection of α-synuclein-pEGFP and pSYNi-1 (transfection group). Cells in all groups were transfected with Lipofectamine 2000 for 48 hours.
MAIN OUTCOME MEASURES: Expression of α-synuclein mRNA and protein were detected by RT-PCR and Western blot. Cell morphology was observed under an inverted fluorescence microscope; cell viability was measured using MTT method; and cell apoptosis was determined with Annexin V-PE flow cytometry.
RESULTS: α-synuclein mRNA and protein expressions were significantly decreased in the pSYNi-1 group when compared with the non-transfect and negative plasmid transfect groups (P 〈 0.05). The expressions were partially decreased in the pSYNi-2 group, but there was no significant difference in the pSYNi-3 and pSYNi-4 groups. Hoechst staining indicated that cell nuclei were enlarged in the negative control group, coloring was not uniform, and chromatin was accumulated and appeared spot-like. The nucleus coloring was uniform in the transfection group compared to negative control group. Cell viability in the negative control group was significantly lower than blank control group with cell apoptosis being significantly increased (P 〈 0.05). In comparison with negative control group, cell viability was significantly increased in the transfection group and cell apoptosis was significantly decreased (P 〈 0.05).
CONCLUSION: pSYNi-1 can inhibit α-synuclein gene expression and block apoptosis of HEK293 cells induced by overexpression of wild-type α-synuclein. 相似文献