结核分枝杆菌Ag85B/GST融合蛋白表达载体构建及在大肠杆菌中的表达

目的 构建结核分枝杆菌Ag85B／GSF融合蛋白表达质粒，并在大肠杆菌(E．coli)中诱导表达。方法 以质粒pUC18／Ag85B为模板，用PCR法扩增结核杆菌Ag85B基因，扩增的Ag85B上游含有EcoR Ⅰ酶切位点，下游含有SalⅠ酶切位点；将Ag85B基因定向插入质粒pGEX-4T-1中，构建原核表达质粒pGEX-Ag85B并转化E．coli B121，筛选阳性重组子，限制性内切酶切鉴定和DNA序列测定，异丙基硫代半乳糖苷(IPTG)诱导表达，SDS-PAGE和Westem blot鉴定结果。结果 成功构建原核表达质粒pGEX-Ag85B并表达出Mr约56000大小的Ag85B／GSY融合蛋白，表达量占总菌体的30％。结论 为进一步进行纯化Ag85B蛋白和研究其在膀胱肿瘤治疗中的作用奠定了基础。     

慢性泪囊炎经鼻腔逆行插管术的护理体会

慢性泪囊炎为眼科常见病之一，需行手术治疗。我科近2年来对86例慢性泪囊炎患者经鼻腔逆行插管术进行了整体护理，取得了良好的效果。现报告如下：     

UVB对角质形成细胞产生细胞因子的影响

角质形成细胞可产生多种细胞因子，这些细胞因子除介导炎症和免疫反应外，还参与组织细胞的生长和分化。UVB照射影响了角质形成细胞合成、分泌细胞因子，并影响细胞因子对角质形成细胞的作用及细胞因子之间的相互作用。概述角质形成细胞产生各种细胞因子的生物学效应。     

Migraine is associated with enhanced arterial stiffness.

Migraine is a common subtype of headache. Epidemiological studies have revealed that migraine could be an independent risk factor for ischemic stroke even in elderly subjects. Arterial stiffness is one of the major pathophysiological bases of stroke. In the present study, we cross-sectionally investigated the possible relationship between migraine and arterial stiffness in community-dwelling subjects. The study subjects were independently recruited from two sources (Group A, n=134, 68+/-5 years; Group B, n=138, 68+/-7 years). Augmentation index (AI), the ratio of augmented pressure by the reflection pressure wave to the pulse pressure, was obtained from the radial arterial waveform as an index of arterial stiffness. Brachial blood pressure was also measured simultaneously. Migraine was diagnosed using a previously validated questionnaire. The prevalence of migraine was 5.2% (Group A) and 16.7% (Group B). Subjects with migraine had higher radial AI in both Group A (migraine, 101+/-15%; other headache, 88+/-12%; no headache, 86+/-12%, p=0.003) and Group B (95+/-11%, 90+/-11%, 91+/-14%, p=0.058). Multiple linear regression analysis revealed that migraine was an independent determinant of AI (beta=0.154, p=0.002) after adjustment for other confounding factors: age (beta=-0.024, p=0.654); sex (beta=0.141, p=0.069); body height (beta=-0.215, p=0.005); systolic blood pressure (beta=0.174, p=0.001); medication for hypertension, hyperlipidemia, and diabetes mellitus (beta=-0.014, p=0.787); and heart rate (beta=-0.539, p<0.001). In a separate analysis by sex, migraine was also a significant determinant for AI (male, beta=0.246, p=0.019; female, beta=0.159, p=0.008). Migraine in the elderly could be a clinical manifestation of enhanced arterial stiffness.     

Temperature dependence of spurious pulses in use of plastic scintillation detectors.

Pulse-height and time spectra of afterpulses from a scintillation detector with NE102A scintillator sandwiching a 60Co source were observed at several different temperatures. From time analysis with a slow amplifier followed by a conventional TAC, spurious pulses are grouped into two types; time-dependent afterpulses and random noise. The intensity of afterpulses decreased considerably with increasing temperature, while the intensity of random noise pulses increased abruptly above 50 degrees C. Experiments, in which the temperatures of scintillator sheets and of the photomultiplier tube were changed separately, suggested that the time-dependent afterpulses were produced in the scintillator itself, while the random component arose from photomultiplier tube.     

Localized wall thickening of the gallbladder mimicking a neoplasm

Clinical diagnosis of chronic cholecystitis is made based on diffuse hyperechoic thickening of the gallbladder wall as shown by ultrasonographic examination. We herein report three cases of chronic cholecystitis showing localized hypoechoic thickening of the gallbladder wall that mimicked gallbladder cancer by ultrasonography. Histologically, hypertrophy of the muscularis propria was a common characteristic finding in these three patients. A smooth surface of the inner hypoechoic layer of the thickened wall was considered to be a reliable finding in the differential diagnosis between this type of chronic cholecystitis and gallbladder cancer.     

健身宝对衰老模型小鼠血清SOD和MDA的影响

目的　研究健身宝补脾益肾、延缓衰老的作用机制。方法　将 60只小鼠随机分为 6组 ,除正常对照组外均每日注射对D -半乳糖造成衰老模型 ,同时给予不同剂量的健身宝和金匮肾气丸。 6周后检测小鼠血清中SOD活性、MDA含量。结果　衰老模型组与正常对照组比较 ,小鼠血清中SOD活性显著降低 ,MDA含量显著升高 (P <0 .0 1 ) ;而健身宝大、中剂量组与正常对照组相比较 ,小鼠血清中SOD活性显著升高 ,MDA含量显著降低 ,与衰老模型组相比较有显著性差异 (P <0 .0 5 )。结论　健身宝明显提升D -半乳糖拟衰小鼠血清中SOD活性 ,降低MDA含量 ,可从调节自由基代谢和提高抗氧化能力方面发挥延缓衰老的作用。     

Nurr1基因转染人脐血间充质干细胞诱导分化为多巴胺能神经元的实验研究

目的探讨外源性Nurrl基因修饰的人脐血间充质干细胞在基因重组成纤维细胞生长因子-8(FGF8)和音猬因子(Shh)诱导下体外分化为多巴胺能神经元的情况。方法间充质干细胞被随机分为A组(对照组)、B组(Nurrl组)、C组(FGF8+Shh组)及D组(Nurrl+FGF8+Shh),采用脂质体法将pcDNA3.1-Ntrr1转染B、D组间充质干细胞,Western blot观察Nurr1基因表达情况,并在神经元条件培养液中进行增殖培养和诱导分化,间接免疫荧光染色法鉴定细胞性质,高效液相色谱法测定多巴胺含量。结果Western blot结果显示转染后Nurr1蛋白表达明显增高。A组和B组未发现酪氨酸羟化酶(TH)阳性细胞,而C组和D组TH阳性细胞分别为10．12％±2．65％及25．36％±3．13％,多巴胺含量分别为(43．6±2．1)nmol／L及(83．2±3．5)nmol／L,差异均有显著性意义(P< 0．01)。结论人脐血间充质干细胞在FGF8和Shh诱导下可以分化为多巴胺能神经元,外源性Nurr1基因修饰后,分化为多巴胺能神经元的数量增加。     

丘脑底核电刺激治疗帕金森病术后程控问题分析

目的 总结丘脑底核电刺激治疗帕金森病术后程控经验，提高术后程控水平，改善疗效。方法 对32例丘脑底核电刺激术后的帕金森病患者进行程控，其中单侧植入者6例，双侧26例；年龄40～73岁，在不同刺激器植入中心接受手术，程控时间术后3周至4年之间。程控前均停药10h以上，程控参数调整主要为刺激电极触点、电压、频率、脉宽四项，程控过程中密切观察病员肌张力和震颤等症状改变以及副反应发生情况，作好详细记录，并分别评估患者程控前后药物“关期”和“开期”症状改善情况，部分行UPDRS评分。结果 31例（97％）患者术后症状得到不同程度改善。刺激电极触点选择中，共29例患者采用单极模式，3例因出现持续、无法耐受的副反应采用双极模式。除1位患者使用循环模式外，其他患者均使用持续刺激模式。刺激电压2．0-4．0V，主要集中于2．8～3-3V，是主要的程控调整参数，电压的高低与病人UPDRS运动评分不具有相关性（P〉0．05）。刺激脉宽60～120μs，刺激频率130～185Hz。药物“关”期，患者UPDRS运动评分，在刺激器打开时，平均18．7分；刺激器关闭时平均47．9分。在刺激器打开情况下，药物“开”期患者症状仍然有进一步缓解．主要表现为步态、全身协调动作方面。合并异动患者6例中，3例适当降低刺激电量，1例提高电压后，异动缓解。结论 丘脑底核电刺激术是有效的帕金森病症状控制手段。准确植入刺激电极是术后获得良好症状控制的前提条件，而术后程控是脑深部刺激器置入术后的关键环节．精确的参数调整能够满意控制病人症状。