哮喘人群STAT6基因多态性的研究

目的 研究信号转导和转录激活因子6（STAT6）基因3’非翻译区G2964A位点多态性与重庆哮喘人群的易感性及血浆IgE水平的关系。方法用聚合酶链反应和单链构像多态性（PCR-SSCP）的方法对42例哮喘患者及42例对照进行了STAT6基因G2964A位点多态性分析。结果哮喘组与对照组STAT6基因G2964A位点的基因型频率之间无显著性差异（P〉0．05），并且哮喘组各基因型之间均与血浆IgE升高无确定关系（P〉0．05）。结论G2964A位点多态性与重庆人群哮喘易感性可能无明显相关性。     

TIM-4基因多态性与湖北地区汉族人群支气管哮喘易感性的研究

目的 探讨T细胞免疫球蛋白域及黏蛋白域蛋白-4(T cells immunoglobulindomain andmucindomain protein-4,TIM-4)基因外显子2区Lys65Lys(G/A)、外显子9区Val1365Met(G/A)的单核昔酸多态性(SNP)与湖北地区汉族人群支气管哮喘易感性的关系.方法 采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)的方法对湖北地区185例哮喘患者和162例健康者TIM-4基因外显子2区Lys65Lys(G/A)、外显子9区Vai365Met(G/A)的多态性进行分析,计算基因型和等位基因频率.结果 (1)湖北地区汉族人群健康者TIM-4基因外显子2区Lys65Lys(G/A)位G/G、G/A、A/A基因型频率分别为0.840、0.160、0,而哮喘人群其频率分别为0.859、0.141、0,其基因型和等位基因型频率与对照组相比差异均无统计学意义(P=0.603,P=0.618);(2)本试验未检测到TIM-4外显子9区Va1365Met(G/A)的多态性.结论 湖北地区汉族人群TIM-4基因外显子2区Lys65Lys(G/A)存在单核苷酸多态性变异,但该位点的变异与湖北地区汉族人群支气管哮喘易感性无关;TIM-4基因外显子9区Va1365Met(G/A)在湖北地区汉族人群中未发现单核苷酸多态性.     

湖北汉族儿童TIM-1基因多态性与变应性哮喘关系的研究

目的　检测湖北地区汉族儿童T细胞免疫球蛋白域粘蛋白域蛋白 1(Tcell,immunoglobulindomainandmucindomainprotein 1,TIM 1)基因第 4外显子插入 /缺失多态性和第 8内含子拼接供体部位(interveningsequence ,IVS) 8 9G/A的单核苷酸多态性 (singlenucleotidepolymorphism ,SNP) ,探讨与儿童变应性支气管哮喘易感性的关系。方法　采用聚合酶链反应检测TIM 1第 4外显子插入 /缺失多态性 ,同时采用PCR 限制性片段长度多态性检测 92名湖北地区健康者和 110例变应性哮喘患儿TIM 1IVS 8 9G/A的SNP ,计算基因型和等位基因频率。结果　 ( 1)湖北地区汉族健康儿童TIM 1第 4外显子缺失 /缺失纯合子、插入 /缺失杂合子和插入 /插入纯合的频率分别是 0 .60 8、0 .3 2 6和 0 .0 65 ;TIM 1IVS 8 9G/G、G/A和A/A的频率分别是 0 .783、0 .196和 0 .0 2 2。 ( 2 )变应性哮喘患儿TIM 1第 4外显子缺失 /缺失纯合子、插入 /缺失杂合子和插入 /插入纯合的频率分别是 0 .63 6,0 3 18,0 .0 45 ,与对照组比较差异无显著性 ,TIM 1IVS 8 9G/G、G/A和A/A的频率分别是 0 .782 ,0 .2 0 9,0 .0 0 9,与对照组比较差异无显著性。结论　湖北汉族儿童存在TIM 1第 4外显子插入 /缺失和第 8内含子IVS 8 9G/A多态性 ,其分布与日本人群相似 ,TI     

E-选择素基因第2外显子G98T单核苷酸多态性的调查

目的 :研究湖北地区汉族人群E 选择素 (E selectin)基因第 2外显子 98位点的单核苷酸多态性 (SNP) ,比较种族间单核苷酸的基因频率分布差异。方法 :应用聚合酶链反应 限制性片段长度多态性(PCR RFLP)的分析方法 ,检测了 2 40名健康者E selectin第 2外显子 98位点单核苷酸的基因型。结果 :E selectin各基因型频率GG型91.3 % ,GT型 8.7% ;G ,T各等位基因频率分别为 95 .6% ,4.4% ,这种基因多态性分布在男女间均无显著性差异 (P >0 .0 5 )。与其它种族比较 ,发现不同种族间E selectin基因型分布及等位基因频率均存在显著差异 (P <0 .0 5 )。结论 :在湖北地区汉族人群中存在E se lectin基因第 2外显子 98位点的单核苷酸多态性 ,这种多态性在种族间可能存在着较大的差异     

瘦素基因-2548A/G和瘦素受体基因Gln223Arg位点多态性与哮喘和代谢综合征的关系

目的:研究瘦素基因-2548A/G和瘦素受体基因Gln223Arg位点多态性与哮喘和代谢综合征的关系。方法:选取82例哮喘合并代谢综合征病人(A+M)、114例哮喘病人(AS)、100例代谢综合征病人(MS)以及96例健康对照者(NC)。根据肺功能将AS组分为轻度AS和中重度AS,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法分析所有受试者瘦素基因-2548A/G和瘦素受体基因Gln223Arg位点单核苷酸多态性及基因型,并比较不同组之间这两个基因位点和多态性之间的差异。结果:1不同组间生化指标有差异(P均<0.05);2瘦素基因-2548A/G位点多态性:MS、A+M和中重度AS组与NC相比AA基因型及A等位基因频率显著升高(P均<0.05);3瘦素受体基因Gln223Arg位点多态性:MS与NC和AS相比AG+AA基因型及A等位基因频率显著升高(P均<0.05),A+M与AS相比A等位基因频率显著升高(P<0.05),AS组、轻度AS组和中重度AS组与NC组相比均没有差异(P均>0.05)。结论:瘦素基因-2548A/G多态性与哮喘和代谢综合征都有一定的相关性,其A等位基因可能是代谢综合征的致病基因,而且与哮喘严重程度相关;瘦素受体基因Gln223Arg A等位基因可能是代谢综合征的致病基因,却与哮喘无关。     

CTLA-4基因多态性与粤西汉族人Graves病遗传易感性的关系

应用限制性片段长度多态性方法检测102例Graves病(Graves'disease,GD)及伴Graves眼病(Graves'ophthalmopathy,GO)亚组患者与100例正常组细胞毒性T淋巴细胞相关抗原-4(cytotoxic T lymphocyte associated antigen-4,CTLA-4)基因外显子1+49位点A/G及启动子-318位点C/T多态性。以探讨CTLA-4基因外显子1+49位点和启动子-318位点多态性与粤西汉族人GD及GO发病的关联性。结果显示GD组外显子1+49位点的GG基因型及G等位基因频率显著高于正常组(P=0.0142、0.0017),GD组AA基因型及A等位基因频率显著低于正常组(P=0.0079、P=0.006);启动子-318位点的各基因型及等位基因频率与正常组相比无统计学意义;外显子1+49位点和启动子-318位点基因型、等位基因频率在GO、无眼病GD亚组及正常组中任两组比较均无统计学意义。研究提示CTLA-4基因外显子1+49位点GG基因型及G等位基因可能是粤西汉族人GD的易感因素,但与GO无相关,AA基因型及A等位基因则是保护因素;启动子-318位点多态性与粤西汉族人GD及GO均不相关。     

一氧化氮合酶3基因多态性与复发性早期自然流产的关系

目的探讨一氧化氮合酶3(nitric oxide synthase 3,NOS3)基因第4内含子27bp数目可变串联重复序列(variable number of tandem repeat,VNTR)多态性和第7外显子894(G/T)多态性与复发性早期自然流产(recurrent early spontaneous abortion,RESA)的相关性。方法选取140例RESA患者和140名健康妇女,应用聚合酶链反应-琼脂糖凝胶电泳法检测NOS3基因第4内含子VNTR多态性,聚合酶链反应-限制性片段长度多态性分析技术检测第7外显子894(G/T)多态性。结果RESA组aa ba基因型频率和a等位基因频率与正常对照组相比差异有统计学意义(χ2=4.51,P<0.05;χ2=4.29,P<0.05)。与bb基因型相比,携带a等位基因的妇女与RESA显著相关(OR为1.8,95%CI:1.04~3.24)。RESA组TT GT基因型频率和T等位基因频率与正常对照组相比差异无统计学意义(χ2=1.16,P>0.05;χ2=1.12,P>0.05)。与GG基因型相比,携带T等位基因的妇女与RESA无相关。结论NOS3基因第4内含子27bp数目可变的串联重复序列多态性与复发性自然流产密切相关,NOS3基因第7外显子894G/T多态性与RESA无明显相关性,a等位基因是RESA重要的遗传易感基因。     

CTLA-4基因多态性与特发性扩张型心肌病的相关性

目的包括细胞和体液免疫在内的自身免疫机制至少参与了部分特发性扩张型心肌病(Idiopathicdilatedcar-diomyopathy,IDC)患者的发病,且前者介导的心肌损害在IDC中更重要。CTLA-4是特异性细胞免疫的负性调节因子。本研究旨在探讨CTLA-4基因启动子-318C/T、外显子A/G多态性及3′非翻译区(AT)n微卫星多态性与IDC及血清可溶性CT-LA-4(sCTLA-4)水平的相关性。方法采用聚合酶链反应-限制性片段长度多态性(Polymerasechainreaction-restrictionfragmentlengthpolymorphisms,PCR-RFLP)方法分析黑龙江省无血缘关系汉族人群(包括72例IDC患者,100例正常健康人)CTLA-4基因-318C/T、49位点A/G多态性及3′微卫星多态性;ELISA法检测血清sCTLA-4水平。综合分析CTLA-4基因型频率、等位基因频率与IDC及sCTLA-4水平的相关性。结果IDC组外显子1GG基因型和G等位基因频率显著高于正常对照组(P=0.012,P=0.008);3′非翻译区共发现18种等位基因,106bp等位基因频率在IDC患者中显著增高(22.22%vs1%,P=0.0002,OR=23.56,95%CI9.65~83.74);两组间-318C/T多态性分布无统计学差异。与对照组相比,IDC组sCTLA-4水平显著升高[(1.87±1.06)μg/L比(0.54±0.19)μg/L,P<0.05];直线回归分析显示,IDC组GG基因型及G等位基因频率与血清sCTLA-4水平(r=0.57,P=0.021)显著相关,而AA、A/G基因型及A等位基因频率与sCTLA-4水平无相关性。启动子-318C/T多态性及3′非翻译区(AT)n微卫星多态性与sCTLA-4水平的亦无相关性。结论CTLA-4基因外显子1A49→G变异与IDC相关,携带G等位基因者易患IDC,其机制可能为该多态性造成CTLA-4信号肽中编码苏氨酸和甘氨酸的替换,从而影响蛋白翻译后加工、修饰,使sCTLA-4功能发生变化。提示3′末端非翻译区(AT)n重复序列中106bp等位基因可能是IDC的易感基因。     

TIM4基因多态性与湖北汉族变应性哮喘的相关性研究

目的分析湖北地区汉族人群T细胞免疫球蛋白域黏蛋白域蛋白4（T cells immunoglobulin domin and mucin domain protein 4，TIM4）基因8570G〉A、11515C〉A单核苷酸多态性，探讨其与变应性哮喘易感性之间的关系。方法采用聚合酶链反应和限制性片段长度多态性对145例变应性哮喘患者和130名健康对照T／M4基因8570G〉A和11515C〉A多态性进行分析，计算基因型和等位基因频率。结果湖北地区健康人群TIM48570G〉AGG、GA和AA基因型频率分别是0．985、0．015和0，而哮喘患者其频率分别为0．931、0．069、0，基因型和等位基因频率在病例组与对照组间差异有统计学意义（P=0．030，P=0．032）；未检测到T／M4基因11515C〉A的多态性。结论湖北汉族人群T／M4基因8570G〉A存在单核苷酸多态性变异，可能与湖北地区汉族变应性哮喘易感性有关。     

Action of dopamine agonists on parkinson-like muscle rigidity induced by 6-aminonicotinamide.

In rats, 6-aminonicotinamide and reserpine induce a Parkinson-like persisting or transient muscular rigidity which was registered electromyographically as spontaneous permanent activity in the gastrocnemic muscle. Seven days after application of 6-aminonicotinamide this pathological finding is already fully developed. At the same time a clear decrease in dopamine concentration in the dopamine-rich striatum and a significant slow-down in the utilization of this transmitter is found. The concentration and utilization of noradrenaline in the diencephalon are unchanged.Both on the 6-aminonicotinamide and on the reserpine model, dose response relations for two dopamine agonists (lisuride and α-bromocriptine) were established by means of electromyography. In both models lisuride was fifty times more effective than α-bromocriptine in reducing the elevated electrical activity. In the 6-aminonicotinamide model, however, 10-fold higher doses of both substances were required to reduce the initial pathological effect by half.The dopamine deficit in the striatum and the effects of l-DOPA and dopamine agonists on the 6-aminonicotinamide-induced muscular rigidity imply that the antimetabolite produces a Parkinson-like syndrome.     

Substance P-like immunoreactivity in nerves associated with the vascular system of guinea-pigs

Substance P-like immunoreactivity was localized by an indirect immunohistochemical technique in whole mounts and sections of blood vessels from the guinea-pig. There was a widespread association of nerve fibres that had substance P-like immunoreactivity with blood vessels, extending into all vascular beds. The relative densities of supply of different vessels were assessed visually and a rating scale used to compare them. Large elastic arteries close to the heart had dense networks of immunoreactive nerves associated with them. The density decreased as more peripheral beds were approached, except that there was a particularly dense network of nerves with arteries of the splanchnic beds. Arteries to myocardial, central nervous system, renal, reproductive and skeletal muscle beds all had substance P-immunoreactive nerves associated with them to varying extents. The venae cavae near the heart were densely supplied, but there were few fibres with their more peripheral extensions. Some large veins (e.g. pulmonary, hepatic portal and superior mesenteric) had a few fibres with them, but veins of peripheral vascular beds had very few or no immunoreactive nerve fibres. Substance P-like immunoreactivity in vascular nerves was markedly reduced in guinea-pigs that were injected with capsaicin but was unaffected by the injection of 6-hydroxydopamine. It is concluded that the vascular substance P-immunoreactive nerves are likely to be of sensory origin.     

The Mer receptor tyrosine kinase is expressed on discrete macrophage subpopulations and mainly uses Gas6 as its ligand for uptake of apoptotic cells

The Mer receptor tyrosine kinase is both an important mediator of apoptotic cell phagocytosis and a regulator of macrophage and DC cytokine production. Since phenotypically distinguishable macrophages are known to have different functions, we have examined Mer expression of murine splenic macrophages. We also used serum deficient in the Mer ligand, growth arrest-specific protein 6 (Gas6) to define better the role of this Mer ligand in macrophage function. By immunofluorescence staining, we found Mer to be strongly expressed in splenic red pulp, largely on platelets. We also found Mer expression on marginal zone macrophages. Strikingly, all tingible body macrophages bore Mer. In functional phagocytosis assays of apoptotic cells, Gas6 appeared to be the sole ligand for Mer, and this system accounted for about 30% of splenic macrophage phagocytosis of apoptotic cells. Taken together, the expression pattern of Mer on macrophage subpopulations in the spleen and its Gas6-dependent role in macrophage phagocytosis suggest an important role for Mer in the modulation of immune responses.     

Interaction between noradrenergic and cholinergic systems in the rat brain: behavioural function in locomotor activity.

Anticholinergic drugs such as scopolamine and atropine induce a mild locomotor stimulation when given intraperitoneally to rats. This effect is usually ascribed to interaction between dopaminergic and cholinergic transmission in the striatum or nucleus accumbens. However, an interaction of acetylcholine with noradrenergic systems is also apparent from biochemical data and the results reported here indicate that at least part of the locomotor activity induced by scopolamine or atropine involves a noradrenergic component. Depletion of forebrain noradrenaline by injection of 4 μg of the selective neurotoxin 6-hydroxydopamine into the dorsal bundle was found to potentiate the locomotor activation induced by various doses of scopolamine or atropine. This was a central effect since methylscopolamine, which does not pass the blood-brain barrier, failed to induce locomotor activity and was not affected by the noradrenergic lesion. The noradrenergic interaction was restricted to cholinergic drugs since locomotor activity induced by the indirect dopamine agonist amphetamine was not affected by noradrenaline depletion.These studies show that the interaction between noradrenergic and cholinergic transmission, which has previously been indicated by biochemical analysis, influences behaviour and they also cast some light on the functions of the central noradrenergic system itself.     

昆明地区儿童G6PD缺乏症临床检测

目的对在我院分娩的新生儿及住院儿童(共5715例)其中男3045人,女2670人,进行G6PD缺乏症筛查.方法采用G6PD/6PGD比值法.结果 144例G6PD阳性,男93人,女51人.男性G6PD缺乏症发生率3.05%,女性发生率为 1.91%.结论广泛开展G6PD缺乏症筛查工作,可使临床医生对G6PD缺乏症有进一步的认识 ,并对患者进行必要的处理,以及在今后的临床用药上起着指导作用.     

昆明地区孕妇夫妇G6PD缺乏症的筛查研究

本文采用G6PD/ 6PGD比值法 ,对 1998年 9月～ 2 0 0 1年 8月来我院门诊就诊的 16 97对孕妇夫妇 (共 3394例 )进行G6PD缺乏症筛查。结果表明 :男性G6PD缺乏症发生率为 3 48% ,女性发生率为 5 30 % ,从而得出昆明地区G6PD缺乏症的基因频率为 0 0 34 8。昆明作为此症的高发地区 ,广泛开展孕妇夫妇G6PD缺乏症筛查工作可使临床医生对缺乏症患者进行必要的处理 ,并帮助她们提高防患意识 ,以避免受到氧化物质的损害 ,从而提高优生优育水平。同时 ,本研究表明 ,G6PD/ 6PGD比值法能较为有效地检出女性杂合子 ,建议广泛应用于临床检测。     

荧光测定法定量测定滤纸片干血斑标本G6PD酶活性的可靠性分析

目的探讨荧光测定法测定滤纸片干血斑标本G6PD酶活性的可靠性。方法随机选取43例门诊孕前咨询者为检测对象,每人采集2ml静脉血抗凝保存同时制备滤纸片干血斑标本。采用G6PD/6PGD比值法测定抗凝血G6PD/6PGD比值,同时采用荧光测定法定量测定滤纸片干血斑标本G6PD酶活性,比较两种方法测定结果。结果荧光测定法检测43份滤纸片干血斑标本,检出1例缺乏（1.57 IU/gHb）;比值法检测43份抗凝血标本,检出1例缺乏（0.76）;两种方法符合率为100%。结论荧光测定法定量测定G6PD酶活性准确性高、简单、快捷、费用低廉,可对滤纸片干血斑标本进行G6PD缺乏症的大规模筛查,适于在G6PD缺乏高发区推广应用。     

小鼠白细胞介素6的cDNA克隆及其在昆虫细胞中的表达

本文采用ＲＴ－ＰＣＲ技术从ＬＰＳ刺激后的小鼠腹腔巨噬细胞，扩增小鼠ＩＬ－６ｃＤＮＡ，并克隆构建ｐＧＥＭ－３Ｚｆ（＋）ＩＬ－６质粒，继将小鼠所得ｃＤＮＡ克隆到ｐＶＬ１３９２载体上，利用杆状病毒ＡｃＮＰＶ表达系统在Ｓｆ９中进行瞬间表达，用ＩＬ－６依赖细胞株ＭＨ６０·ＢＳＦ２检测其表达活性，结果表明，感染后４８ｈ后就具有明显ＩＬ－６表达，由此可见，利用该系统可成功地表达小鼠ＩＬ－６基因。     

The KSHV viral interleukin-6 is not essential for latency or lytic replication in BJAB cells

Kaposi's Sarcoma-associated herpesvirus encodes a homolog of the human cellular interleukin-6 that may play a formative role in many KSHV-related diseases. While the viral IL-6 can signal similarly to its human counterpart little is known about the role of vIL-6 during KSHV infection. Using homologous recombination and selection in eukaryotic cells, a KSHV isolate was purified that does not express vIL-6 as was a control recombinant that left vIL-6 intact. The two viruses establish and maintain latency to similar levels in BJAB B-cells, reactivate to similar levels in B-cells and Monkey kidney cells and have very similar KSHV gene expression patterns. BJAB cells expressing KSHV survive better than the parental BJAB cells in low serum and the vIL-6 deletion does not abrogate this growth advantage. Thus vIL-6 is not essential for establishment, maintenance, or reactivation from latency in cell culture and is not involved in the survival of infected BJAB B-cells in low serum.