SARS病毒S蛋白编码基因表达载体的构建及在Vero细胞中的表达

目的构建含SARS病毒S蛋白编码基因片段的表达载体,并将其置于Vero细胞中表达。方法设计1对PCR引物,在其下游引入Flag序列,通过PCR方法从质粒pGEX-6P-1-SARS-S中扩增出S蛋白编码基因,PCR产物经酶切后,插入表达载体pcDNA3.1(+)中,构建重组表达质粒pcDNA3.1(+)-SARS-S。重组质粒经酶切鉴定及核苷酸序列测定和分析后转染Vero细胞,用抗Flag单克隆抗体通过Western blot检测S蛋白片段在Vero细胞中的表达。结果酶切鉴定及核苷酸序列测定和分析示重组质粒构建完全正确;Western blot证实重组S蛋白片段能够在Vero细胞表达。结论融合Flag序列的SARS病毒S蛋白片段在Vero细胞中获得了正确表达。     

红霉素联合莫沙比利治疗急性脑卒中患者胃肠运动障碍的疗效观察

急性脑卒中伴发胃肠运动障碍十分常见,主要表现为胃排空延缓而导致的腹胀、呕吐、返流,并容易导致吸入性肺部感染,严重影响预后.本研究采用红霉素联合莫沙比利治疗急性脑卒中伴发胃肠运动障碍,效果良好,现报道如下.     

Hp感染胃疾病与Treg细胞及褪黑素的关系

幽门螺杆菌(Helicobacter pylori,Hp)定植可引起胃黏膜炎症反应,是导致十二指肠溃疡和胃癌的主要危险因素之一。全球约一半人口感染Hp,我国Hp感染率更高达50%-80%,其中约10%-20%的感染者将发展为胃溃疡或胃癌。新近研究发现Hp感染患者机体中调节性T细胞(Regulatory T cell,Treg细胞)增多,后者能抑制机体对Hp的过度免疫应答和炎症反应,在介导免疫耐受形成和炎症慢性化过程中可能起重要作用,并可促进胃癌的发生和发展。褪黑素(Melatonin,MLT)对胸腺Treg细胞的产生有显著调控作用,并呈剂量依赖性。相关研究资料表明,MLT与Treg细胞、与Hp、与胃癌之间存在一定的功能联系。但目前国内外有关MLT与Hp感染所致胃疾病直接联系的研究较少,本文就Hp感染-Treg细胞-褪黑素之间的关系做一综述。     

针、药、灸三法治疗脑卒中后尿失禁106例分析

目的研究脑卒中后尿失禁患者尿动力学异常以及针刺、艾灸和中药联合应用对脑卒中后尿失禁的治疗效果。方法脑卒中后尿失禁患者106例,行尿动力学检查,分析膀胱尿道功能;并将患者随机分为治疗组与对照组,对照组给予脑卒中常规治疗,治疗组在脑卒中常规治疗的基础上,给予针刺、艾灸和中药联合治疗。结果脑卒中后尿失禁主要尿动力学异常为逼尿肌反射亢进和尿道外括约肌无抑制性松弛,其主要病变部位为额叶和基底节。针刺、艾灸和中药联合治疗能明显改善脑卒中后尿失禁症状。结论脑卒中后尿失禁患者尿动力学改变以逼尿肌反射亢进和尿道外括约肌无抑制性松弛为主;针刺、艾灸和中药联合治疗脑卒中后尿失禁疗效显著。     

经尾静脉注入大鼠骨髓间充质多能成体祖细胞向脑缺血灶微环境中的移行及效应

BACKGROUND： Multipotent adult progenitor cells （MAPCs） from the bone marrow have been shown to differentiate into neurons.
OBJECTIVE： To observe migration, survival, and neuronal-like differentiation of MAPCs by tail vein injection. DESIGN, TIME AND SETTING： Randomized, controlled experiment of neural tissue engineering was performed at the Laboratory for Cardio-Cerebrovascular Disease, Hospital of Integrated Traditional and Western Medicine, Tongji Medical College of Huazhong University of Science and Technology between September 2006 and August 2007. MATERIALS： Eighty Sprague Dawley rats, 3-6 months old, underwent cerebral ischemia/reperfusion by thread technique, and were randomly divided into model and MAPCs groups （n = 40）. METHODS： Mononuclear cells were harvested from bone marrow using the FicolI-Paque density gradient centrifugation method. After removing CD45 and glycophorin A-positive cells （GLYA＋） with immunomagnetic beads, CD45 GLYA adult progenitor cells were labeled with bromodeoxyuridine （5-bromo-2-deoxyuridine, BrdU）. A total of 1 mL cell suspension, containing 5 × 10^6 MAPCs, was injected into the MAPCs group through the tail vein. A total of 1 mL normal saline was injected into the model rats.
MAIN OUTCOME MEASURES： After 60 days, BrdU and neuron-specific enolase double-positive cells were observed using immunofluorescence. Cell morphology was observed under electron microscopy, and nerve growth factor mRNA was measured through RT-PCR. In addition, rat neurological functions were measured with behavioral tests.
RESULTS： Immunofluorescence revealed that MAPCs positive for BrdU and neuron specific enolase were found surrounding the ischemic focus in the MAPCs group. Microscopic observation suggested that MAPCs-derived neuronal-like cells connected with other nerve cells to form synapses. Compared with the model animals, the level of nerve growth factor mRNA was significantly upregulated in rats injected with MAPCs （P 〈 0.05）. In addition, rats in the MAPCs group performed better in behavioral tests than the model group on days 28 and 60 （P 〈 0.05）.
CONCLUSION： Transplanted MAPCs migrated to the ischemic region, survived, and differentiated into neuronal-like cells, resulting in stimulation of nerve growth factor mRNA and improved neurological function in ischemic rats.     

卵巢切除与血清肿瘤坏死因子-α、转化生长因子-β1浓度的关系

目的探讨卵巢摘除与血清肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)浓度的关系. 方法 30只SD大鼠随机分为模型Ⅰ(OVXⅠ)组、模型Ⅱ(OVXⅡ)组和假手术组,OVXⅠ、OVXⅡ组大鼠卵巢切除,术后12和24周运用酶联免疫法检测大鼠血清中TNF-α、TGF-β1浓度的变化. 结果大鼠去卵巢后当雌二醇(E2)水平下降时,血清TNF-α浓度[(53.88±16.35)ng/L]升高,而TGF-β1含量[(6.78±0.75)μg/L]下降,与假手术组比差异有显著性(P＜0.05或P＜0.01);相关性分析显示,E2与TNF-α呈负相关(r=-0.384,P=0.05),而与TGF-β1正相关(r=0.750,P=0.001),两细胞因子之间亦呈负相关(r=-0.465,P=0.05). 结论 E2对TNF-α有抑制作用,但却能促进TGF-β1的分泌,TNF-α含量的增加和TGF-β1水平的降低,均能刺激骨的吸收活动,导致骨质疏松症的发生.     

针灸、微波配合盆底肌锻炼治疗脑卒中后尿失禁106例分析

目的：分析脑卒中后尿失禁患者尿动力学异常情况以及针灸、微波配合盆底肌锻炼治疗脑卒中后尿失禁的效果。方法：选择2004—07／2006—05在华中科技大学同济医学院附属中西医结合医院神经内科住院脑卒中后尿失禁患者106例，随机分为治疗组59例与对照组47例，对照组给予脑卒中常规治疗，治疗组在脑卒中常规治疗的基础上，给予针刺、艾灸以及微波配合盆底肌锻炼等综合治疗。分别于治疗前及治疗30d后行尿动力学检查，分析膀胱尿道功能。结果：106例患者均进入结果分析。①两组患者最常见的尿动力学异常为逼尿肌反射亢进、尿道外括约肌无抑制性松弛，治疗组占37％（22／59），对照组占41％（19／47），两组比较差异无显著性（P〉0．05）。②脑卒中性质和病灶部位对患者尿动力学结果无显著影响（P〉0．05）。③治疗组经综合治疗后尿动力学检查结果较治疗前明显改善[膀胱最大收缩压：（5．16&;#177;2．04），（6．65&;#177;2．43）kPa；膀胱最大容积：（363．50&;#177;194．14），（317．11&;#177;159．72）mL；尿道闭合压：（8．53&;#177;3．23），（5．23&;#177;2．17）kPa；膀胱充盈初始量：（174．73&;#177;105．32），（188．30&;#177;84．95）mL，P〈0．05]。④治疗组显效率和有效率明显高于对照组（显效率：26％，53％；有效率：80％，71％，P〈0．05）。结论：脑卒中后尿失禁患者尿动力学改变以逼尿肌反射亢进和尿道外括约肌无抑制性松弛为主；针刺、艾灸、微波配合盆底肌锻炼治疗脑卒中后尿失禁效果显著。     

喜普妙治疗脑卒中后情绪障碍的疗效分析

目的探讨脑卒中后患者存在的情绪障碍、喜普妙对脑卒中后情绪障碍的治疗效果及其对患者生活质量的影响。方法采用心理量表对1700例卒中单元住院患者进行心理评估。将存在情绪障碍的脑卒中患者随机分为治疗组及对照组，评定与分析两组患者SCL-90、HAMD、HAMA、SF-36的分值及其临床疗效。结果76％脑卒中患者存在一定程度的情绪障碍，以抑郁、焦虑症多发。治疗组HAMD、HAMA评分显著低于对照组，其临床疗效及生活质量评分显著高于对照组。结论脑卒中后患者容易出现情绪障碍；喜普妙能显著改善脑卒中患者的情绪障碍，可以提高临床疗效、患者的躯体功能及生活质量。     

雌孕激素与子宫内膜的一氧化氮合酶

NO(NitricOxide) /NOS(NitricOxideSynthase)在雌性生殖生理功能方面如卵泡发育、激素分泌、胚胎着床、维持妊娠、促进分娩及内膜周期性变化、细胞凋亡等方面都具有重要作用。有关子宫内膜NOS的表达及调控机制仍存在异议。三种NOS神经型NOS(neuronalNOS ,nNOS)、内皮型NOS(endothelialNOS ,eNOS)和诱导型NOS(inducibleNOS ,iNOS)在内膜不同部位和时期呈强弱不等的表达 ,生理作用也不尽相同 ,且与雌孕激素对内膜的调节 ,内膜周期性变化关系密切。本文就NOS在子宫内膜的活性表达及其与雌孕激素关系的研究进行综述。     

Nimodipine Modulates Bcl-2 and Bax mRNA Expression after Cerebral Ischemia

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA.The increased ratio of Bcl-2 and Bax mRNA may contribute to the antiapoptic effect of nimodipine.The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new stratehgy to manage neuronal damage.