CTLA-4基因多态性在重症肌无力发病机理中的作用

目的探讨细胞毒性T淋巴细胞相关抗原-4(cytotoxicTlymphocyteassociatedantigen-4,CTLA-4)基因第1外显子 49位点、启动区-318、-1661、-1772位点的多态性及其导致的无效转录对重症肌无力(myastheniagravis,MG)遗传易感性的影响。方法酶联免疫吸附实验测定MG患者和健康对照血清中可溶性CTLA-4的水平;限制性片段长度多态性分析检测第1外显子 49位点、启动区-318、-1661、-1772位点的多态性;转录因子核因子(nuclearfactor1,NF-1)和CCAAT/增强子结合蛋白β(CCAAT/enhancerbindingproteinbeta,c/EBPβ)结合位点通过染色质免疫沉淀实验得以验证。结果启动区-1772、-1661位点和第1外显子 49位点的多态性与MG,特别是伴发有胸腺瘤的MG密切相关。启动子-318位点的多态性与MG无关。CTLA-4基因4个多态性位点间有一个明确的正性连锁不平衡关系。MG患者血清可溶性CTLA-4的表达水平与等位基因的突变相关联。-1772、-1661位点的多态性可改变转录因子NF-1和c/EBPβ结合位点,而ConA、PHA则能促进NF-1和c/EBPβ的这种位点特异性转录活性。结论MG患者CTLA-4A/G 49、C/T-1772和A/G-1661多态性可导致无效转录,影响MG的遗传易感性,T→C-1772的突变能影响基因的剪接,从而干扰蛋白的表达和功能。     

CTLA-4基因启动子区单核苷酸多态性与特发性扩张型心肌病的关联研究

目的探讨特发性扩张型心肌病（idiopathic dilated cardiomyopathy，IDC）患者细胞毒性T淋巴细胞相关抗原-4（cytotoxic T lymphocyte assoccated antigen 4，CTLA-4）表达状况及由CTLA-4基因启动子区单核苷酸多态性（single nucleotide polymorphism，SNP）导致的不同遗传易感性机制。方法采用限制性片段长度多态性分析151例IDC患者，120名正常健康人CTLA-4基因启动区-1772、-1661及-318位点SNP；免疫酶联吸附测定法检测血清sCTLA-4、干扰素-7及白介素-4水平；综合分析CTLA-4启动区基因型、等位基因频率及与sCTLA-4、干扰素-γ／白介素一4的相关性。结果IDC患者sCTLA-4水平与CTLA一4基因启动区SNP相关，携带-1772T／C变异者sCTLA-4表达增高。-1772TC基因型频率在IDC组尤其低射血分数亚组显著高于对照组，IDC组-1661G和-1661GG频率显著降低，具有-1772TC-1661AA及-1772TC-1661AG单倍型IDC患者sCTLA-4显著升高。结论IDC患者CTLA-4表达异常，CTLA-4基因启动区-1772C／T和-1661A／GSNP与IDC遗传易感性相关。-1772T／C变异可能影响CTLA-4基因剪接，干扰蛋白表达和功能，阻止负性调节信号传递而导致对IDC的易感。     

CTLA-4基因多态性调节溃疡性结肠炎患者CTLA-4 mRNA稳定性和蛋白水平

目的 研究细胞毒T淋巴细胞相关抗原4(cytotoxic T-lymphocyte associated antigen 4,CTLA-4)基因3'非转录区(AT)n重复序列多态性对溃疡性结肠炎(ulcerative colitis,UC)患者CTLA-4mRNA稳定性和基因表达的影响.方法 采用实时定量PCR方法检测膜型CTLA-4(full length CTLA-4,flCTLA-4)和可溶性CTLA-4(soluble CTLA-4,sCTLA-4)mRNA表达,半衰期法分析其mRNA稳定性.免疫组化检测flCTLA-4蛋白表达.酶联免疫吸附实验测定sCTLA-4蛋白水平.荧光PCR-毛细管电泳技术检测300例UC患者和700名健康对照者CTLA-4基因(AT)n重复序列多态性.结果 活动期UC患者肠黏膜sCTLA-4 mRNA的表达显著低于缓解期UC患者(P=0.004).在UC患者中,(AT)n重复序列长等位基因携带者表达低水平的flCTLA-4和sCTLA-4 mRNA以及sCTLA-4蛋白(均P＜0.01).携带长等位基因的UC患者CTLA-4 mRNA的稳定性明显降低.UC患者CTLA-4基因(AT)n重复序列长等位基因携带者(≥116 bp)频率显著高于正常对照组(P＜0.01),且与广泛型结肠炎相关(P=0.008).结论 UC患者CTLA-4基因(AT)n重复序列多态性与CTLA-4基因表达水平相关,携带(AT)n重复序列长等位基因的UC患者CTLA-4 mRNA及蛋白的表达降低,提示CTLA-4基因在UC遗传免疫发病机制中起重要作用.     

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。     

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。     

细胞毒T淋巴细胞相关抗原4基因多态性与乳腺癌易感性的关联研究

目的: 探讨CTLA-4基因多态性位点-1722T/C和CT60G/A与中国北方汉族妇女乳腺癌易感性的关系.方法: 采用聚合酶链反应(PCR)和限制性片段长度多态性方法, 对328例中国北方汉族乳腺癌患者和327例正常对照者进行CTLA-4基因-1722位点和CT60位点多态性检测. 结果: 乳腺癌患者CTLA-4基因CT60位点G等位基因频率在乳腺癌患者组中明显高于正常对照组(28.7% 比23.5%; P=0.0352, OR=1.30, 95% CI=1.02～1.67); -1722C-CT60A单体型在对照组中的频率大于病例组中的频率, 有明显差异(P=0.0283, OR=0.77, 95% CI=0.97～0.61), 而在-1722位点基因型频率、等位基因频率与对照组比较差异无统计学意义(P＞0.05).结论: CTLA-4基因多态性-1722和CT60两个位点与我国北方汉族妇女乳腺癌发病存在一定的相关性.     

特发性扩张型心肌病CTLA-4基因3''非翻译区(AT)n微卫星多态性研究

目的 特发性扩张型心肌病（idiopathic dilated cardiomyopathy，IDC）的发病机制与T细胞免疫应答密切相关。细胞毒性T淋巴细胞相关抗原-4（cytotoxic T lymphocyte antigen-4，CTLA-4）主要在已激活的T细胞上表达，通过与CIY28竞争结合B7，抑制T细胞过度激活，维持免疫系统内环境稳定。CTLA-4基因3’非翻译区（AT）n微卫星多态性影响CTLA-4功能。本研究旨在探讨外周血单个核细胞（PBMC）CTLA-表达状况及3’非翻译区（AT）n微卫星基因多态性与IDC的相关性。方法分别用原位杂交、免疫组化、序列特异性引物PCR等方法检测38例无血缘关系的北方汉族IDC患者以及50例正常对照者的CTLA-4mRNA、蛋白质表达、CTLA-4基因外显子3’末端非翻译区（AT）n重复序列多态性，并对PCR扩增产物进行序列分析。结果IDC组与对照组相比，PBMC经金黄色葡萄球菌肠毒素B刺激后CTLA-4mRNA、蛋白质表达强度显著减弱，且无一定规律；3’末端非翻译区共发现18种CTLA-4等位基因，与对照组比较，106bp等位基因频率在IDC患者中显著增高[22．22％ vs 1％，P=0．0002．OR=23．56，95％可信区间（CI）：9．65～83．74]。结论IDC患者CTLA-4基因转录和表达缺陷，该缺陷与CTLA-4基因3’末端非翻译区（AT）n重复序列多态性存在关联。     

TNF-α基因多态性与妊娠高血压的相关性研究

目的　探讨肿瘤坏死因子-α(tum or necrosis factor- alpha,TNF-α)基因启动子- 30 8G>A、- 85 0 C>T多态性与妊娠期高血压疾病的相关性。方法　应用聚合酶链反应-限制性片段长度多态性技术检测10 6例患者和10 8名健康孕妇的TNF- α基因启动子- 30 8G>A、- 85 0 C>T多态性。对两组之间的基因型频率和等位基因频率进行比较。结果　TNF-α基因启动子- 30 8位点TNF2等位基因频率和TNF2 / 1基因型频率在病例组明显升高(P<0 .0 5 )。- 85 0位点T等位基因频率和CT+TT基因型频率在对照组明显升高,差异有统计学意义(P<0 .0 5 )。由这两个多态性位点组成的不同基因型中,病例组TNF2 / 1CC基因型频率明显高于对照组,差异有统计学意义(P<0 .0 5 ) ,而TNF1/ 1TT基因型频率在对照组明显增高,差异有统计学意义(P<0 .0 5 )。结论　TNF- α- 30 8、- 85 0位点多态性与妊娠期高血压疾病相关,其中TNF- α基因- 30 8位点的突变是危险因素,- 85 0位点的突变可能是保护性因素。TNF2 / 1CC基因型可能是妊娠期高血压疾病的易感基因型。     

TMX基因多态性与先天性肥厚性幽门狭窄易感性的传递不平衡检验分析

目的 探索TMX基因外显子6上的多态性位点rs7161242[ c.492T＞G]及外显子7上的多态性位点rs7160810[ c.648 G＞A]与先天性肥厚性幽门狭窄(CHPS)发病易感性的关联.方法 对广州市第一人民医院收治的22个汉族核心家系(CHPS患者及父母)采用PCR及测序的方法进行基因分型.应用传递不平衡检验(TDT)判断基因多态性与CHPS发病的关联.结果 测序结果未发现新的突变位点;患儿及父母组内这两个多态性位点的Hardy-Wcinberg平衡检验均P＞0.05.TDT检验提示多态性位点rs7161242的G等位基因及rs7160810的A等位基因均与CHPS发病相关,其p值分别为2.0×10-4和5.699x10-5.连锁不平衡分析结果提示,这两个位点的r2为0.757,D′值为0.893,成紧密连锁.结论 TMX基因的多态性位点rs7161242[ c.492T＞G]及rs7160810[c.648 G＞A]与中国汉族人群CHPS发病密切相关.     

血清可溶性 CTLA-4表达与重症肌无力的相关性研究

目的探讨血清可溶性细胞毒性T淋巴细胞相关抗原-4（CTLA-4）表达与福建地区重症肌无力（ myasthenia gravis ,MG）患者的相关性。方法 ELISA法检测福建地区80例MG患者（ MG患者分为激素未治疗组及激素治疗组,且激素治疗组进一步分为免疫抑制剂治疗组和胸腺切除组）和80例正常对照组血清可溶性CTLA-4（ sCTLA-4）水平。结果 MG激素未治疗组、激素治疗组的sCTLA-4均高于正常对照组[（6．03±3．58） ng/ml、（3．44±2．36） ng/ml vs （0．49±0．95） ng/ml],它们之间的差别具有统计学意义（χ2＝100．67,P＜0．001）;其中激素治疗组sCTLA-4表达水平高于正常对照组（Z＝-9．03,P＜0．001）,而 MG激素未治疗组sCTLA-4表达高于激素治疗组（Z＝-3．37,P＝0．001）;并且激素未治疗组激素治疗前后血清sCTLA-4水平差异有统计学意义（t＝3．10,P＝0．005）;胸腺切除术后sCTLA-4低于手术前（Z＝-2．21,P＝0．04）,而免疫抑制剂治疗前后差异却没有统计学意义（Z＝-1．26,P＝0．21）。结论 sCTLA-4参与MG的发病机制,激素治疗、胸腺切除治疗减少sCT-LA-4表达。     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Changes in the hypothalamic nuclei in in experimental allergy

Summary In rabbits subjected to prolonged sensitization and in which the Arthus phenomenon was induced there was a marked reaction of the hypothalamic nuclei. Staining by Gomori's method indicated a cellular swelling, loss of granules, and protoplasmic vacuolization in the supraoptic nucleus. There was a considerable increase in the size of the cross-sectional area of the cells. The same effects were much less well shown in the paraventricular nucleus. These results show that marked signs of increased neurosecretion developed in the animals at the height of the Arthus phenomenon.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten Éksperimental'noi biologii i Meditsiny, Vol. 55, No. 4, pp. 110–113, April, 1963.     

Experience in adult population in dengue outbreak in Delhi

A dengue outbreak has recently hit the Indian capital. We studied the clinical profile of adult patients. Five hundred and sixty patients of dengue infection were admitted in a specially created ward according to the criteria laid down by WHO. Haematemesis (28.28%), epistaxis (26.78%) and malena (14.28%) were some of the common presentations. Similarly lymphadenopathy, especially cervical (30.89%), palatal rashes (26.96%) and hepatomegaly (23.75%) were the most commonly encountered findings on physical examination. Most of the cases were of dengue fever with haemorrhage and only 2.5% cases were classified under dengue haemorrhagic fever or dengue shock syndrome. The average hospital stay was 3.4 days but only 9.8 hours in the eleven patients who died, suggesting their late arrival in preterminal situation giving little time for resuscitation. Thrombocytopenia was not a feature and only 12.85% patients had platelet count less than 70,000/cmm. Most of the patients who were admitted with thrombocytopenia, showed normalization in their platelet counts in next few days. Serological examination demonstrated evidence of recent dengue infection in 41.17% patients. Few patients required blood or platelet concentrate transfusion. Eleven patients died, three due to DIC, one of intracranial haemorrhage and seven due to massive gastric haemorrhage. Rest of the patients recovered completely. Thus we can conclude that recent outbreak in Delhi was of dengue fever with haemorrhage and mortality was very low in patients who came early to the hospital.     

Intestinal helminthiasis in school children in Haiti in 2002

A survey on intestinal helminths in school children was conducted in Haiti in 2002. This first nationwide study involving the entire country was stratified by department according to urban and rural zones using the cluster method. Focusing on elementary school children (n=5792; age range 3 to 20 years), it involved 26 urban and 49 rural schools randomly selected. Stools were preserved in formalin and examined by the Ritchie technique. Thirty-four per cent of stools (1981/5792) tested positive for intestinal helminths with the following parasites identified: Ascaris lumbricoides (27.3%), Trichuris trichiura (7.3%), Necator americanus (3.8%), Hymenolepsis nana (2%), Taenia sp. (0.3%) and Strongyloides stercoralis (0.2%). The helminth prevalence was higher in rural (38.4%) compared to urban areas (30%). There was no significant difference in prevalence by sex and age. The importance of geohelminths changed from one department to another with the highest prevalence found in the Southern department of Grande Anse (73.7%) and the lowest prevalence in the Center department (20.6%). Five out of the country's nine departments had a similar prevalence varying from 25.5% to 28.2%. Intestinal helminthic polyparasitism was observed in a percentage of infested school children comprise between 3.4% and 28.6% according in relation to the geographical area. A program to fight against geohelminths in school children should be initiated as a public health priority. Albendazole is the drug of choice. Frequency of drug distribution should be based on the prevalence of geohelminths in each department.     

Cerebellar noradrenergic systems in aging: studies in situ and in in oculo grafts

Age-related changes in noradrenergic function in the rat cerebellum were examined using electrophysiological and electrochemical techniques. Sprague-Dawley and Fischer 344 rats showed subsensitivity to norepinephrine (NE) locally applied onto cerebellar Purkinje neurons. The modulatory actions of NE on Purkinje cell-evoked activity was also examined. In young rats NE preferentially inhibits spontaneous activity more than evoked excitations when compared to control. These modulatory actions of NE are not seen in senescent Fischer 344 rats. The intrinsic vs. extrinsic influences determining the loss of efficacy to NE were examined using three groups of rats with in oculo cerebellar grafts. The first group had young grafts grown in young hosts and these grafts showed a potent response to perfused NE. The second group, old grafts in old hosts, showed a diminished responsiveness to NE with respect to the first group. The third group consisted of young grafts in old hosts. These grafts demonstrated a responsiveness to NE that was indistinguishable from those in the first group. The integrity of the presynaptic NE fibers was examined in the grafts using electrochemical techniques. No difference in the release of NE was observed in the old grafts. Taken together, these results suggest a loss of postsynaptic NE function that is intrinsically determined. The change in NE modulation could influence information processing within the aged cerebellar cortex. This deficit could underlie behavioral changes seen in senescence.