人TAT-PDX-1融合蛋白在大肠杆菌中的表达与纯化

目的构建人TAT-PDX-1表达载体,并对其融合蛋白进行纯化。方法采用RT-PCR技术从人胰腺组织中获得目的基因hPDX-1,将TAT序列与PDX-1克隆到原核表达载体pET-28a;用IPTG诱导、表达融合蛋白;NiNTAagaros纯化融合蛋白,Western blot鉴定。结果目的片段PDX-1被有效扩增,DNA序列测定表明所构建重组质粒pET28a-TAT-h PDX-1与设计相同;TAT-PDX-1融合蛋白在大肠杆菌中获得高效表达并成功纯化。结论成功地获得了TAT-hPDX-1融合基因的表达产物,为进一步研究及临床应用奠定了基础。     

超选择动脉溶栓治疗80岁以上脑梗死的安全性及疗效评估-两中心研究

Objective To evaluate the feasibility, safety and efficacy of intra-arterial thrombolytic therapy on elderly patients (≥ 80 years old) with acute ischemic stroke. Methods The clinical data of 86 patients with acute ischemic stroke, received intra-arterial thrombolytic therapy, were retrospectively analyzed; according to age differences, these patients were divided into advanced age group (≥80 years old, n=21) and common age group (<80 years old, n=65); and control group (≥80 years old, not receiving thrombolytic therapy, n=50) was established. The recanalization rate and early clinical improvement rate, and the incidence, recover rate and death rate of symptomatic intracerebral hemorrhage were evaluated in these patients after treatment. Results No significant differences in the favorite recanalization rate and short-term outcome, and the incidence of symptom intracranial hemorrhage were noted between the advanced age group and common age group (P=0.528, P=0.102,P=0.353). The incidence of symptom intracranial hemorrhage in the advanced age group was obviously higher than that in the control group (P=0.034); the recover rate of symptom ntracranial hemorrhage in the advanced age group (42.9%) was obviously lower than that in the common age group (50.8%), but significantly higher than that in the control group (16%, P=0.042, P=0.017). The mortality of the advanced age group was similar to that of the control group (23.8% versus 28%, P=0.816), but higher than that of common age group (23.8% versus 10.8%, P=0.034). Conclusion Relatively high feasibility, safety and efficacy of intra-arterial thrombolytic therapy are noted in elderly patients (≥80 years old) with acute ischemic stroke, demonstrating that the use of intra-arterial thrombolytic therapy in very elderly patients should not be avoided but pursued advisably.     

实验性脑室出血大鼠脑脊液S100β含量的变化

目的 观察脑室出血大鼠脑脊液S100β含量的变化.方法 采用大鼠脑室出血模型,应用酶联免疫吸附方法测定脑脊液S100β含量.结果 大鼠脑室出血后24 h,脑脊液S100β浓度显著升高.结论 脑脊液S100β可作为脑室出血后脑组织损伤的敏感标志物. Abstract： Objective To observe the changes of cerebrospinal fluid(CSF) S100β concentrations in experimental ventricular hemorrhage rat model.Methods By using enzyme-linked immunosorbent assay, the concentration S100β in CSFprepared from experimental ventricular hemorrhage model rats were determined.Results CSF S100β concentrations in model rats were significantly higher than that of normal and saline-control rats,after 24 hours of ventricular hemorrhage.Conclusions CSF S100β concenfrations can be a sensitive marker of ventricular hemorrhagic brain damage.     

TA-SOD融合蛋白表达载体的构建

目的:构建pET28a-TAT-SOD原核表达载体,纯化并测定其活性.方法:PCR法扩增TAT-SOD片段,插入载体pET28a后得到pET28a-TAT-SOD重组表达子,转化人肠杆菌BL21(DE3),IPTG诱导TAT-SOD融合蛋白表达.表达产物用SDS-PAGE电泳鉴定,组氨酸亲和层析柱纯化融合蛋白,利用SOD活性测定试剂盒测定其活性.结果:构建了高表达重组子pET28a-TAT-SOD,获得了相对分子质量约为25 000的具有活性的融合蛋白TAT-SOD,纯度达90%.纯化的蛋白浓度为700 mg/L,活性为197.54 U/L.结论:成功构建了具有活性TAT-SOD融合蛋白的表达载体.     

高同型半胱氨酸血症患者个体化治疗疗效分析

高同型半胱氨酸血症(hyperhomocysteinemia,HHcy)作为脑血管病的重要危险因素<'[1-2]>,其治疗方法国际上尚无统一意见[3].目前,国内一般建议使用叶酸、维牛素B<,6>或B<,12>联合治疗,但适用范围、治疗时间长短缺乏明确规定.而临床实际中,多数HHcy患者仅伴有一种维生素的缺乏.为此,作者实施了改进的个体化治疗方案,即依据患者体内叶酸、维乍素B<,12>的不同水平,有针对性地补充患者所缺乏的特定维生素,取得了较好的临床效果,报道如下.     

携带人脱嘌呤/脱嘧啶核酸内切酶( APE1)基因重组慢病毒载体的构建及体外表达检测

目的:构建携带人脱嘌呤/脱嘧啶核酸内切酶-1 (Apurinic/apyrimidinic endonuclease-1,APE1)基因重组慢病毒载体内英文摘要中,并检测其体外表达目的基因的水平.方法:应用基因克隆技术将人APE1基因克隆入慢病毒载体LV中,经过PCR、酶切和测序鉴定后,与包装四质粒系统共转染293T细...     

河南汉族人群血小板膜糖蛋白I ba基因Kozak序列多态性与缺血性脑血管病的关联性

目的:探讨河南地区汉族人群血小板膜精蛋白(GP)I ba Kozak序列基因多态性与缺血性脑血管病的关联性.方法:采用PCR-RFLP方法对缺血件脑血管病组(228例)和健康对照组(235例)进行Kozak序列多态性分析.结果:缺血性脑血管病组CC基冈型频率为8.8%,C等位基因频率为29.2%,均高于对照组的4.3%和23.2%(X2=3.896,P=0.048;X2=4.281,P＝0.039).结论:GP I ba基冈Kozak序列多态性可能与河南汉族人群缺血性脑血管病的发生相关.     

脑干梗死的危险因素分析

目的探讨脑干梗死的危险因素。方法根据脑梗死部位将193例脑梗死患者分为大脑半球梗死组(128例)和脑干梗死组(65例),比较两组间性别、年龄、吸烟、合并症、血脂及糖化血红蛋白(HbA1c)水平,以及两组患者基底动脉和椎动脉狭窄度、基底动脉弯曲分级、椎动脉优势的比率;采用Logistic回归方法分析脑干梗死危险因素。结果与大脑半球梗死组比较,脑干梗死组年龄明显降低,吸烟、合并糖尿病的比率明显增高,血低密度脂蛋白及HbA1c水平明显增高(P<0.05～0.01);基底动脉和椎动脉狭窄率、基底动脉弯曲≥2级率及椎动脉优势比率明显增高(P<0.05～0.01)。Logistic多因素回归分析显示,伴有糖尿病(OR=4.018,95%CI:1.796～8.992;P=0.0006)、基底动脉狭窄(OR=1.003,95%CI:1.018～1.048;P<0.0001)和基底动脉弯曲≥2级(OR=1.377,95%CI:1.008～1.059;P=0.0087)与脑干梗死的发生独立相关。结论伴有糖尿病、基底动脉狭窄和基底动脉弯曲度是脑干梗死的重要的高危因素。     

缺血性脑血管病患者蛋氨酸合成酶基因A2756G多态性检测

目的:检测缺血性脑血管病患者蛋氨酸合成酶(MS)基因A2756G多态性.方法:运用PCR-限制性片段长度多态性(RFLP)技术对512例缺血性脑血管病患者及500例健康对照者进行MS基因多态性检测.结果:缺血性脑血管病患者组MS A2756G A/A、A/G、G/G基因型频率分别为88.9%、10.7%和0.4%,A和G等位基因频率分别为94.2%和5.8%.对照组A/A、A/G、G/G基因型频率分别为84.6%、15.0%和0.4%,A和G等位基因频率分别为92.1%和7.9%.2组基因型及等位基因频率差异均无统计学意义(P＞0.05).结论:MS基因A2756G多态可能不足以构成河南汉族人群缺血性脑血管病发病中的一个独立的遗传危险因子.     

脊髓受累的可逆性后部脑病综合征的临床特征及预后

目的探讨脊髓受累的可逆性后部脑病综合征(PRES-SCI)的临床表现、影像特征及预后。方法分析郑州大学第一附属医院神经内科2021年11月收治的1例PRES-SCI患者的临床资料, 并检索国内外数据库, 对已报道的38例PRES-SCI患者及本例患者进行汇总分析。结果 39例PRES-SCI患者(包括本例)主要临床表现为头痛(79.5%, 31/39)、视力障碍(79.5%, 31/39)、呕吐(46.2%, 18/39)、意识障碍(38.5%, 15/39)、肢体无力(28.2%, 11/39)、癫痫发作(23.1%, 9/39)。97.4%(38/39)的患者有血压明显升高。影像特征性表现为以中央灰质受累为主的长节段脊髓病变, 89.7%(35/39)的脊髓病灶起源于延髓颈髓交界处, 病变累及椎体所对应的脊髓节段数中位数为9(最短为4, 最长为全脊髓)。38例合并颅脑病变, 最常累及的部位为延髓(82.1%, 32/39)、枕叶(46.2%, 18/39)、脑桥(43.6%, 17/39)、顶叶(41.0%, 16/39)、小脑(38.5%, 15/39)。38例完成随访的患者中,...