SERPINA3K原核表达系统的构建及其表达

目的：通过构建丝氨酸蛋白酶抑制因子SERPINA3K的原核表达载体,并将其转化到Rosetta-g ami2感受态细胞中,表达重组SERPINA3K,从而为对其功能的深入研究奠定了基础.方法：以小鼠MS-1细胞cDNA为模板通过PCR的方法扩增Serpina 3k基因的表达全片段,再将其插入到原核表达载体pET-41a中,酶切并测序鉴定重组体.将构建好的重组质粒转化大肠杆菌Rosetta-gami2感受态细胞,表达产物用Western blot鉴定.结果：原核表达载体pET-4 1a-Serpina 3k成功构建,可在大肠杆菌Rosetta-gami2中高效表达,得到重组蛋白SERPINA3K,经Western blot鉴定正确.结论：成功构建SERPINA3K原核表达载体且获得表达,为研究SERPINA3K生物学活性及产品开发提供了实验基础.     

PMA诱导节律基因表达对细胞紫外线损伤的影响

目的研究诱导NIH3T3细胞节律基因表达对细胞紫外线损伤的影响。方法用体外节律诱导剂乙酰肉豆蔻佛波酯诱导NIH-3T3细胞节律基因表达。分别选择mPer2基因表达的谷值和峰值时刻对细胞进行紫外线照射,以非诱导节律的单纯照射组为对照,通过流式细胞术检测细胞凋亡和细胞周期分布,平板克隆形成试验检测细胞增殖情况。结果mPer2基因表达峰值时进行紫外线照射的细胞,与mPer2基因表达谷值时和非诱导节律紫外线照射细胞比较,增殖能力强、凋亡率低、G1期阻滞明显。结论节律基因诱导的NIH3T3细胞的紫外线损伤减小,并与mPer2的基因表达相关,其保护作用可能与mPer2基因过表达有关。     

PITX3 polymorphism is not associated with Parkinson's disease in a Chinese population

Several studies have indicated that three PITX3 single nucleotide polymorphisms (SNPs), rs2281983, rs4919621 and rs3758549, are likely to be associated with Parkinson's disease (PD) in Caucasians. Some studies also suggested an age-of-onset effect. We recently reported that allele and genotype frequencies did not differ between late-onset PD (LOPD) patients and controls for all three SNPs. To extend the analysis to early-onset PD (EOPD) patients, and to test whether an age-of-onset effect exists in Chinese, we genotyped these SNPs in 290 Chinese EOPD patients using a ligase detection reaction (LDR). For all three SNPs, allele and genotype frequencies did not differ between total PD patients and controls, between LOPD patients and controls, between EOPD patients and controls, or between LOPD and EOPD patients. Our results suggest that these PITX3 SNPs do not contribute to the risk of developing PD in EOPD or LOPD in Chinese.     

中国汉族人群近日钟基因与高脂血症的关联

目的 针对中国汉族人群,开展Clock、Clif节律基因多态性与高脂血症间关联研究,以进一步探讨近日节律对高脂血症的影响与作用,为基于易感基因的靶向药物治疗提供新思路.方法 采用病例-对照关联分析（casecontrol association study）,以明确诊断的205例高脂血症患者和281例健康对照者为研究对象,研究节律基因相关的基因位点与高脂血症易感性、疾病严重程度等临床表型之间的关系并阐明其机制.根据选取的节律基因位点Clock（rs3840267,rs3749474,rs10529257）和Clif（rs2289709）,确定相关位点的基因型,采用遗传统计学分析确定基因型与临床表型之间的关联性.结果 通过Clock、Clif基因多态性与高脂血症的病例-对照分析,发现Clock rs 10529257等位基因显著增加了高脂血症的患病风险（8.51％ vs 13.95％,Paalleo=0.003,ORalle.=0.529,95％CI=0.346,0.809）;Clockrs3840267、rs3749474和Clifrs2289709在病例组和对照组中无显著差异（P＞0.05）.通过高脂血症分型后病例-对照分析,发现Clock rs 10529257在Ⅳ型高脂蛋白血症亚型病例组和对照组之间有显著差异（9.72％vs 13.95％,Palleo-Ⅳ=0.043,ORalleoⅣ =0.525,95％CI=0.278,0.991）;Clif rs2289709在Ⅱb型高脂蛋白血症亚型病例组和对照组之间亦有显著差异（19.23％vs8.72％,Palleo-Ⅱb=4.738±10-4,ORalleo-Ⅱb=2.959,95％CI=1.645,5.322）.通过病例组中总胆固醇、甘油三酯、低密度脂蛋白胆固醇水平与两个阳性位点Clock rs 10529257、Clif rs2289709不同基因型比较发现,Clock rs10529257中-/A杂合子携带者与A/A纯合子基因型携带者的甘油三酯水平存在显著差异[（1.27±0.69） vs（1.76±1.22）,P=0.01];Clif rs2289709中A/G杂合子携带者与G/G纯合子基因型携带者的甘油三酯、低密度脂蛋白分别存在显著差异[（2.31±1.81） vs （1.11±0.32）,P=2.33×10^-8;（2.83±0.98） vs （1.63±0.69）,P=5.83×10^-15）.结论 本研究结果发现,节律基?     

Circadian learning and memory changes in Aβ1–42 induced Alzheimer’s mice

Metabolic Brain Disease - Alzheimer disease (AD) is a growing health problem globally, which causes a progressive decline in learning and memory and multiple disturbances of circadian rhythms. Six...     

A comparative study of lesionectomy versus corticectomy in patients with temporal lobe lesional epilepsy

We performed a retrospective study comparing surgical outcome of anterior temporal lobectomy (n = 20) and stereotactic lesionectomy (n = 14) in 34 consecutive patients with intractable partial epilepsy evaluated at this institution between June 1986 and June 1992. All patients had medically refractory partial seizures and a neuroimaging-identified temporal lobe foreign-tissue lesion. Patients underwent a pre- and postoperative comprehensive evaluation. In all patients, postoperative neuroimaging studies indicated complete lesion resection. Seventy-one percent of lesionectomy patients and 90% of lobectomy patients, respectively, experienced a worthwhile reduction in seizure tendency. Forty-three percent of lesionectomy patients and 85% of lobectomy patients, respectively, were seizure-free postoperatively. Mean duration of follow-up was 51 months (range, 21–83 months). Morbidity occurred in three patients who underwent corticectomy: complete homonymous hemianopsia (1) and dysphasia (2). No significant morbidity was associated with lesionectomy. Results of this study have altered the surgical strategy at this institution for patients with temporal lobe lesional epilepsy and have proved useful in counseling patients regarding neurologic outcome.     

吗啡成瘾小鼠对正常小鼠学习记忆力能力的影响

目的:用BALB/C小鼠建立动物模型,观察吗啡成瘾戒断小鼠对其周围的正常小鼠学习记忆能力的影响。方法:将吗啡成瘾戒断小鼠与正常小鼠同笼饲养,然后用Morris水迷宫实验测定正常小鼠的学习记忆能力,应用RT-PCR的方法检测正常小鼠海马中PKCa、GABA分子表达的变化。结果:水迷宫实验证明,与生理盐水组相比,实验组小鼠的学习记忆能力明显降低(P<0.05),海马中PKCa和DABA的表达都明显升高(P<0.05)。     

慢性吗啡成瘾小鼠海马中RACK1对CREB表达水平的影响

目的：研究shRAKC1对慢性成瘾小鼠脑组织CREB mRNA、蛋白的表达变化。方法：通过条件性位置偏爱（CPP）实验分析shRAKC1对慢性吗啡成瘾小鼠的作用;通过RT-PCR检测RACK1和CREB的mRNA在成瘾小鼠海马中的表达水平,并采用免疫组化观察RACK1和CREB蛋白的表达情况。结果：慢性成瘾组与生盐水组相比,前者海马区RACK1和CREBmRNA表达升高（p〈0．05）,且吗啡诱导的CPP效应增强（p〈0．05）,而shRAKC1组与空质粒组相比,前者由吗啡诱导的CPP诱导效应减弱（p〈0．05）,同时其海马区RACK1和CREBmRNA表达下降（p〈0．05）。结论：干扰慢性吗啡成瘾小鼠RACK1表达,可以使CREB表达水平下调,并有抑锏吗啡成瘾效应的作用。