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采用简并引物克隆葎草花粉过敏原全长同源cDNA 总被引:1,自引:0,他引:1
目的:建立一套稳定可靠的方法,对过敏原性物种中的过敏原同源基因进行快速克隆。方法:在分析生物信息数据库中积累的大量过敏原序列同源性的基础上,设计简并引物,基于高质量Lǘ草花粉RNA,逆转录合成cDNA。采用Touchdown方式在cDNA池中进行选择性PCR扩增。同时借助梯度PCR程序,对引物扩增的简并性作进一步强化,并结合RACE技术获取全长cDNA.进而对Lǘ草花粉中的过敏原同源基因进行克隆。结果:成功地获得3个全长cDNA克隆。序列分析显示.这些基因与已知过敏原的基因序列相似性高达79%-85%,初步认定其为泛过敏原肌球蛋白抑制蛋白(profilin)的同源基因。对比RACE技术获得的相应基因的全长序列发现,这些序列在引物结合处与简并引物序列之间存在4个碱基的差异,提示采用Touchdown方式的梯度PCR程序.可使引物的简并性得到进一步扩展。结论:简并引物与Touchdown梯度PCR相结合的方法.能够对以Lǘ草为代表的基因组未曾深入研究的物种中的过敏原同源基因进行有效克隆. 相似文献
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Objective
To observe the effect of acupuncture in regulating ubiquitin-proteasome pathway (UPP), and discuss the action of acupuncture in intervening heroin-induced brain damage.Methods
Thirty male Sprague-Dawley (SD) rats were divided into a control group, a model group and an acupuncture group by using the random number table. Rats in the model and acupuncture groups received intramuscular heroin injection for successive 8 d at a progressively increased dose. Afterwards, the injection was suspended for 5 d for withdrawal. The heroin relapse rat model was established by repeating the drug addiction and withdrawal process for 3 times. The control group followed the step of the model establishment, but was given intramuscular injection of normal saline at the stage of addiction and no intervention at the stage of withdrawal; the model group was given intramuscular heroin injection at a progressively increased dose at the addiction stage and no intervention at the withdrawal stage; the acupuncture group was dealt in the same way as the model group at the addiction stage, but received acupuncture at Baihui (GV 20) and Dazhui (GV 14) at the withdrawal stage, with the needles retained for 30 min each time, 1 session a day, for successive 5 d. On the 39th day, brain tissues were extracted from the hippocampus and ventral tegmental area (VTA) of the three groups of rats. The apoptosis of brain nerve cells was detected by using terminal deoxynucleotidyl transferase-mediated nick and labeling (TUNEL). The mRNA and protein expressions of ubiquitin (Ub), ubiquitin protein ligase (E3) and 26S were examined by immunohistochemistry and quantitative real-time polymerase chain reaction (RT-qPCR).Results
Compared with the model group, rat’s hippocampus and VTA in the acupuncture group showed significantly fewer cells positively stained by TUNEL staining (P<0.01), and its mRNA and protein expressions of Ub, E3, 26S were significantly lower (P<0.01).Conclusion
Reducing nerve cell apoptosis and regulating the mRNA and protein expressions of Ub, E3 and 26S in rat’s hippocampus and VTA are possibly one of the action mechanisms of acupuncture in intervening heroin-induced brain damage.3.
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背景缺血性脑卒中发病率、死亡率、致残率均较高,溶栓后的再灌注损伤对患者影响较大,针刺是治疗本病的特色疗法,但作用机制尚不明确。目的 探讨“通督调神”电针预处理对脑缺血再灌注损伤(CIRI)大鼠miR-124-3p/糖原合成酶激酶β(GSK-3β)/亲环素D(Cyp-D)信号通路及线粒体膜通透性转换孔(MPTP)的影响,探讨其防治CIRI的可能机制。方法 2022年6—8月,将100只清洁级SD大鼠随机分为假手术组、模型组、电针组、抑制剂组和电针+激动剂组,每组20只。造模前干预7 d,电针组、电针+激动剂组选取通督调神穴组:百会、风府、大椎穴进行电针干预,1次/d,连续7 d;造模前24 h电针+激动剂组和抑制剂组分别侧脑室注射miR-124激动剂和抑制剂(5 nmol)。除假手术组,余组采用改良线栓法制备大鼠右侧脑缺血再灌注模型;造模成功后,取大鼠右侧脑皮质,采用改良神经功能损伤评分量表(mNSS)、TTC染色观察各组大鼠神经功能损伤程度,TUNEL染色以及透射电镜观察神经细胞损伤情况;免疫荧光染色、Western blotting、实时荧光定量PCR检测各组大鼠脑皮质GSK-3β、... 相似文献
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浅谈医院档案工作的管理 总被引:3,自引:0,他引:3
医院的档案,反映了医院工作的历史和现状,又是医院工作的依据和参考。在医院的全面管理工作中,特别是在医疗质控和服务质量的管理及基本建设方面,档案是历史和法律的凭证。 相似文献
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微小核糖核酸(microRNA,miRNA)是一类进化上高度保守的非编码中链小分子RNA,能够参与调节脑缺血再灌注损伤(cerebral ischemia reperfusion injury, CIRI)病理生理过程,被认为是潜在的脑缺血(cerebral ischemia, CI)诊断生物标志物。miR-124是大脑皮层和小脑中优先表达的miRNA,在缺血性脑损伤发生后,miR-124与人体内多个靶点结合,通过调控细胞凋亡、自噬、神经炎症、氧化应激反应、促进神经保护与再生、抑制兴奋性氨基酸毒性等多种机制调节缺血性脑损伤的发展走向,具有一定的神经恢复潜力,且在调控细胞凋亡、神经炎症的过程中具备负向调节作用。但目前有关miR-124调控缺血性脑损伤在临床层面的证据相对匮乏,需要在广泛的临床试验中进一步论证其作为缺血性脑损伤诊断生物标志物、治疗靶点的可能性。 相似文献
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背景:“通督调神”电针预处理法是结合传统针刺和现代电刺激的新兴物理治疗方法,既往文献报道其对缺血性脑卒中具有显著治疗作用,但其具体机制尚不明确。目的:探讨“通督调神”电针预处理作用于细胞凋亡,对大鼠缺血半暗带区miR-124和皮质区Notch-1、p-JNK和半胱氨酸天冬氨酸特异性蛋白酶3蛋白相对表达量的影响及其可能的作用机制。方法:将75只雄性SD大鼠随机分为模型组、假手术组、电针预处理组、miR-124抑制剂预处理组和电针+miR-124激动剂预处理组,每组15只。7 d干预期内,电针预处理组取“风府”“百会”“大椎”穴行电针,1次/d,20 min/次;miR-124抑制剂预处理组第7天注射miR-124antagomir至大鼠侧脑室内;电针+miR-124激动剂预处理组第7天在电针基础上注射miR-124 agomir;模型组及假手术组不予治疗。干预结束后造模,假手术组只做鼠板上固定和钝性分离血管,其余组均采用大脑中动脉闭塞方法并参照Zea Longa线栓法建立大鼠右侧脑缺血再灌注损伤模型。造模完成后24 h应用改良神经损伤程度评分定损,评分结束后使用TUNEL染色法测细胞凋亡... 相似文献
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目的 观察针刺人迎穴为主联合加味牵正汤治疗急性期周围性面瘫(PFP)的疗效。方法 将80例急性期PFP患者随机分为两组,均予西医常规治疗,对照组40例针刺常规取穴联合内服加味牵正汤,观察组40例另增加针刺人迎穴治疗。比较两组治疗前后House-Brackmann(H-B)分级、面部残疾指数(FDI)[(躯体功能(FDIp)和社会功能FDIs)]、症状和体征积分的变化。结果 观察组总有效率为95.00%,显著优于对照组的82.51%(P <0.05)。治疗后,观察组患者H-B分级、FDIs评分、症状和体征积分均低于对照组(P <0.05),FDIp评分高于对照组(P <0.05)。结论 针刺人迎穴为主联合加味牵正汤能有效改善急性期PFP患者的临床症状,提高生活能力。 相似文献
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