蛇毒Ⅱ类磷脂酶A2功能分化的分析研究

目的：研究蛇毒Ⅱ类磷脂酶A2(PLA2)中D49 PLA2和K49 PLA2的功能分化及其功能分化决定位点的鉴定。方法：运用序列比较分析，进化树构建和DIVERGE v1．04软件计算研究D49 PLA2和K49 PLA2的功能分化情况及其分化位点。结果：序列比较分析，进化树构建和DIVERGE v1．04软件计算结果表明蛇毒Ⅱ类PLA2中D49 PLA2和K49 PLA2的确发生了功能分化，对于K49 PLA2来说，1S，7K，11Q，E12，R34，T56，N88，L92，E108，K116，K128可能为功能分化决定位点。对于D49 PLA2，L2，G33，G35，F46和Y118可能为功能分化决定位点。结论：我们首次通过序列比较分析，进化树构建和DIVERGE v1．04软件计算鉴定出蛇毒Ⅱ类PLA2中D49 PLA2和K49 PLA2可能的功能分化位点，为今后通过基因重组和定点突变方法研究蛇毒Ⅱ类PLA2结构功能关系提供了线索。     

幽门螺杆菌感染及其毒力因子和胃十二指肠疾病关系的研究

目的 探讨幽门螺杆菌（Hp)感染病人中Hp细胞毒相关蛋白（CagA)、细胞空泡毒素（VacA)抗体在胃十二指肠疾病的比例，并评估不同类型幽门螺杆菌感染对胃窦粘膜炎症程度的影响。方法 应用免疫印迹法测定Hp感染者病人体内的CagA、VacA抗体。结果 73％的Hp感染者病人中出现CagA抗体，61％Hp感染者病人中出现VacA抗体。CagA抗体和VacA抗体在各疾病之间差异无显著性（P＞0．05）。在胃溃疡、十二指肠溃疡和复合性溃疡三类病人中，具CagA和VacA双阳性的比例较CagA和VacA双阴性的比例差异有显著性（P＜0．05）。胃窦萎缩病变在CagA和VacA双阳性的病人中比CagA和VacA双阴性的病人严重（P＜0．05）。结论 具有高毒力因子（CagA和VacA)的Hp是胃肠疾病病人感染的常见菌株，具有CagA和VacA的Hp可能在诱导胃粘膜发展到萎缩病变过程中起重要作用。但不能作为特异性指标来判断胃十二指肠疾病。     

Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues

BACKGROUND: The mucosal mast cell (MMC) granule-specific beta-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-beta1 induces constitutive release of mMCP-1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated. OBJECTIVE: To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2. METHODS: MMC homologues were generated by culturing bone marrow cells in the presence of TGF-beta1, IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi-disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA. RESULTS: mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited ( approximately 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2. CONCLUSION: The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.     

重组人白细胞介素10的融合表达及鉴定

目的 研究重组人白细胞介素10(rhIL—10)载体在大肠杆菌B121(DE3)pLyse细胞中的表达，为进一步研究IL-l0在动脉粥样硬化中的作用机制奠定基础。方法 用构建成功的IL-l0—PCRT7/NT-TOPO质粒转化大肠杆菌BL21(DE3)pLyse细胞，并通过SDS-PAGE鉴定融合表达蛋白。结果 PCRT7/NT—TOPO质粒载体成功载入rhIL-l0基因；在异丙基硫代—β-D-半乳糖苷(IPTG)诱导下表达的蛋白质主要以包涵体形式存在。结论 在IPTG诱导下，重组的IL-l0—PCRT7/NT—TOPO质粒载体在大肠杆菌BL2l(DE3)pLyse细胞内成功表达。     

高效液相色谱法测定石杉碱甲片的含量

采用高效液相色谱法测定石杉碱甲片的含量。固定相为μＢｏｎｄａｐａｋ Ｃ１８，流动相为乙腈－０．０２ｍｏｌ／Ｌ磷酸二氢钾溶液（１２：８８），以对乙酰氨基酚为内标，石杉碱甲浓度在１０￣５０ｍｇ／Ｌ范围内与峰面积呈良好的线性关系，平均回收率为９９．９３％，ＲＳＤ为０．４１％（ｎ＝９）。     

硒和维生素A对支原体肺炎的治疗效果观察

观察硒和维生素A(VA)对支原体肺炎的治疗效果.方法 采用双盲随机对照2×2析因实验设计,选择100例住院支原体肺炎患儿,分为补硒组26例,补VA组23例,补硒和VA组30例以及病例对照组21例,正常组21例.一次补硒量为1mg亚硒酸钠和/或15万单位VA,对照病例组给予常规治疗.结果 与对照组比,治疗后3个补充组的症状和体征缓解天数均有不同程度缩短(P<0.05),补硒组白细胞硒和谷胱甘肽过氧化物酶水平显著上升(P<0.05),补VA、补硒组血清VA水平上升(P<0.01),细胞免疫功能有所改善.结论 硒和VA有协同作用,补充硒或同时加VA,作为辅助治疗支原体肺炎的方法,安全、有效.     

Association of angiotensin II type 1 receptor gene polymorphism with essential hypertension

The renin-angiotensin system is involved in control of blood pressure and salt and fluid homeostasis. Genes for components of this system have been of major focus in research on the causation of the common, complex, polygenic trait, essential hypertension (HT). Association of an A→C variant at nucleotide 1166 of the angiotensin II type 1 receptor (AT₁R) gene with HT, but an absence of linkage of this locus with this disease, has been reported recently. Since confirmation in a different setting is imperative, we performed a cross-sectional case-control study of the A1166C variant in a well-characterized group of 108 Caucasian HT subjects with a strong family history (two affected parents) and early onset disease. Genotyping was by mismatch polymerase chain reaction/ Bfr I restriction fragment length polymorphism analysis. Frequency of the C¹¹⁶⁶allele was 0.40 in HTs and 0.29 in normotensives. The difference in genotype (χ²= 13, P = 0.0015) and allele (χ²= 5.3, P = 0.02) frequencies between the two groups was significant (odds ratio for CC vs AA+AC = 7.3 [95% CI, 1.9–31.9). The present results implicate the AT₁R gene, or a locus in linkage disequilibrium with the variant tested, in the causation of essential HT.     

人体β-胡萝卜素的肠转化和吸收后转化的研究

目的 :为了解部分中国人体内 β 胡萝卜素 ( β carotene ,以下简称 β C)转化维生素A (vitaminA ,以下简称VA)的效率 ,开展了本研究。方法 :使用稳定同位素稀释法对 15名 5 0 60岁健康农村志愿者 (男 9,女6)进行β C人体代谢实验。 2周适应期和 5 6天实验期内 ,志愿者接受常规膳食 ,避免大量VA和 β C摄入以及烟、酒和营养补充剂。实验第 1天 ,给受试者含 6mg氘标记 β C( 2 H8β C)玉米油胶丸 ,随半流质早餐 (脂肪热能比 2 5 % )一起摄入。实验第 4天 ,受试者以同样方法摄入含 3mg氘标记醋酸视黄醇 ( 2 H8RAC)油剂胶丸。实验第 1天和第 4天摄入标记物后 0 ,3 ,5 ,7,9,11,13h时 ,实验第 2 ,3 ,5 ,6,7,8,9,10 ,14,2 1,2 8,3 5 ,42 ,49,5 6天晨空腹时 ,采静脉血。用高效液相色谱仪 (HPLC)分离血清 β C和VA组分 ,再分别使用气相质谱仪(GC MS)和液相质谱仪 (LC MS)测定VA和 β C组分的同位素丰度。根据VA和 β C的浓度和同位素丰度 ,描述标记VA和β C在体内应答的血液动力学曲线。 结果 :所有 15名受试者对2 H8RAC应答明显 ;但是在对2 H8β C的应答方面 ,只有 11名受试者2 H4视黄醇应答曲线明显 ,有 4名受试者血清2 H4视黄醇应答曲线非常微弱。经过对备份血清样品进行的多次重复GC MS测定 ,我们目前初步?     

Utilization of care in haemophilia: a resource-based method for cost analysis from the Haemophilia Utilization Group Study (HUGS)

Summary.  The Haemophilia Utilization Group Study (HUGS) was created 10 years ago to examine the annual utilization and cost of haemophilia-related healthcare services. Retrospective chart reviews for 336 patients with haemophilia A receiving treatment in one of five comprehensive haemophilia treatment centres (HTCs) during 1995 were completed through interview of the provider. This method provided adequate collection of data from patient charts without the abstractor having direct access to patient health information. Utilization data were used to impute the costs of different components of care (e.g. physician visits, factor VIII concentrate, emergency room, hospitalization). The total annual cost of care was $139 102 (SD $304 033). Factor VIII concentrate costs comprised the largest proportion of these costs; mean factor VIII concentrate use was 128 517 units per patient per year. Unbilled physician utilization accounted for 7.8% of the mean total physician costs per annum, while mean allied healthcare costs accounted for 33.5% of the total annual allied healthcare costs per patient. In the ordinary least-squares regression model, higher costs were associated with severe factor VIII deficiency, arthropathy, more comorbid conditions, an inhibitor to factor VIII concentrate, infusing through a port and prophylaxis. Although factor VIII concentrate is the most costly component, the treatment of haemophilia uses many healthcare resources. HUGS has demonstrated that patient clinical characteristics and physician practices predominantly drive the costs of haemophilia care. Specifically, patients with more severe arthropathy had greater healthcare costs. As future funding decisions are made, it is important to provide for all components of care.