214例脑梗死患者与HPA-1～5、15基因多态性关系的检测与分析

目的:探讨老年易感病脑梗死与HPA-1~5、15基因多态性的相关性。方法:采用PCR-SSP技术对214例60岁以上脑梗死病例组和220例健康体检者对照组进行HPA-1~5、15系统基因分型,并应用统计学方法对两组进行χ~2检验和Logistic回归分析。结果:1 HPA-2抗原系统多态性病例与对照组比较差异有统计学意义。脑梗死组基因频率(a=0.915 9,b=0.048 1)与对照组基因频率(a=0.977 3,b=0.022 7)相比较P0.01;脑梗死组基因型频率(aa=0.831 8,ab=0.168 2,bb=0)与对照组基因型(aa=0.9545,ab=0.0455,bb=0)相比较P0.01。2 HPA-2回归分析:脑梗死HPA-2b基因和HPA-2ab基因型均显著高于对照组,以HPA-2aa为指示符进行Logistic回归分析并以年龄调整后,HPA-2ab的OR值=4.123(95%CI=1.981~8.581)。结论:HPA-2抗原系统基因多态性可能是本地区老年人群脑梗死发病的遗传易感因素,HPA-2b可能是其易感基因,HPA-2ab可能是其易感基因型。     

血小板膜糖蛋白Ⅰa和Ⅰbα基因多态性与脑梗死的相关性研究

目的探讨血小板膜糖蛋白(GP)Ⅰa基因和Ⅰbα基因多态性与脑梗死发生的关系,为缺血性脑卒中的预防及治疗提供理论基础。方法选择经CT或MRI证实的脑梗死患者302例(脑梗死组)和健康体检者196例(对照组);采用PCR-RFLP方法检测GPⅠa C807T基因与Ⅰbα基因HPA-2、Kozak序列多态性在2组中的分布频率。结果脑梗死组GPⅠa C807T等位基因频率明显高于对照组,差异有统计学意义(P0.05);脑梗死组GPⅠbα基因Kozak序列C等位基因频率明显高于对照组差异有统计学意义(25.33% vs 10.20%,P0.05);脑梗死组GPⅠbα基因HPA-2序列等位基因频率、基因型与对照组比较,差异无统计学意义(P0.05)。结论 GⅠbα基因HPA-2序列多态性与脑梗死无相关性;GPⅠa C807T等位基因和Ⅰbα基因Kozak序列多态性可能是脑梗死的遗传危险因素。     

GpⅡb／Ⅲa基因多态性与冠心病关系的研究

目的探讨血小板膜糖蛋白（Gp）Ⅱh／Ⅲa基因多态性与冠心病的关系。方法选择冠心病患者86例（A组）和健康体检者80例（B组）,运用聚合酶链-序列特异性引物技术进行GpⅡb人类血小板抗原3（HPA-3）及GpⅢaHPA-1多态性检测。结果A组和B组中GpⅡb HPA-3aa、ab、bb基因型分布有统计学差异,A组ab、bb基因型的分布较B组高,A组b等位基因频率高于B组。GpⅡb HPA-3ab和bb基因型在心肌梗死组中的分布、b等位基因频率高于非心肌梗死组。结论GpⅡb HPA-3b等位基因可能是冠心病的遗传易患因子,GpⅡb HPA-3可能为心肌梗死发生的遗传易感性标志,GpⅢa HPA-1基因可能与蔻心病发生无关联。     

胱硫醚β合酶基因T833C突变与脑梗死发生的相关性研究

目的探讨脑梗死患者同型半胱氨酸(Hcy)代谢相关酶胱硫醚β合酶(CBS)基因T833C位碱基突变与脑梗死发病的相关性。方法对67例经头部CT或MR I证实为脑梗死的患者(脑梗死组)和31名健康对照者(对照组),应用聚合酶链反应(PCR)扩增法检测CBS基因T833C多态性,采用高效液相色谱法测定血清Hcy水平。结果在脑梗死组有9例CBS基因纯合子突变,28例为杂合子突变;对照组3名为纯合子突变,5名为杂合子突变。两组基因型频率分布差异亦有显著意义,2χ=11.429,P<0.01;脑梗死组C等位基因频率为34.33%,T等位基因频率为65.67%,与对照组比较差异有显著意义,2χ=8.978,P<0.01。CBS基因杂合突变者血清Hcy浓度显著高于正常基因者(t=4.612,P<0.01)。脑梗死组患者血清Hcy浓度为(23±7)nmol/m l,显著高于对照组(13±4)nmol/m l,两组比较差异有显著性(t=8.826,P<0.01)。结论Hcy血症是脑梗死的独立危险因素,而CBS基因T833C点突变可能是其发病的重要遗传因素。     

血小板膜糖蛋白Ⅱb HPA-3基因多态性与中国汉族人缺血性卒中的相关性

目的 探讨天津市汉族人群血小板膜糖蛋白(glycoprotein,GP)Ⅱb HPA-3基因多态性的分布情况及其与缺血性卒中的关系.方法 缺血性卒中组150例,对照组135例,应用聚合酶链反应-限制性内切酶片段长度多态性方法进行基因分型,并抽样测序验证.比较各基因型和等位基因频率分布及其与缺血性卒中的关系.结果 病例组ab基因型、bb基因型和b等位基因频率均显著高于对照组(P =0.000),而aa基因型和a等位基因频率则显著低于对照组(P=0.000).病例组不同性别和年龄组之间GPⅡb基因型和b等位基因频率无显著性差异;虽然各病因学亚型之间基因型频率无显著性差异,但大动脉粥样硬化性卒中亚组b等位基因频率显著高于小血管闭塞性卒中亚组和心源性脑栓塞亚组(61.8％对46.7％对47.5％x2 =6.573,P=0.037).多变量logistic回归分析显示,高血压(优势比7.475,95％可信区间3.700 ～ 15.003;P=0.000)和b等位基因(优势比3.678,95％可信区间1.245～10.863,P=0.018)是缺血性卒中发生的独立危险因素.结论 血小板膜GPⅡb HPA-3基因多态性可能与缺血性卒中发生风险相关,携带b等位基因可能是缺血性卒中的独立危险因素,尤其是大动脉粥样硬化性卒中.     

基质金属蛋白酶-1基因多态性与老年脑梗死患者颈动脉粥样硬化程度的相关性

目的探讨基质金属蛋白酶(MMP)-1基因多态性与老年脑梗死患者颈动脉粥样硬化程度的相关性。方法选取120例老年颈动脉粥样硬化性脑梗死患者和120例健康体检者,脑梗死患者根据颈动脉超声检查结果分为不稳定斑块组(60例)和稳定斑块组(60例),运用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测各组MMP-1基因-519A/G的多态性,采用酶联免疫吸附法测定各组外周血MMP-1水平,比较其基因型及等位基因分布频率和各组外周血MMP-1水平的差异。结果梗死组AA基因型分布频率(70.00%)及A等位基因的分布频率(82.08%)明显高于对照组(55.83%和74.58%)(χ2=5.16、χ2=1.65,P均<0.05),梗死组MMP-1血清含量明显高于对照组(t=18.45,P<0.05);不稳定斑块组和稳定斑块组外周血MMP-l浓度比较差异无统计学意义(t=0.31,P>0.05),不稳定斑块组MMP-1基因-519A/G位点AA基因型的分布频率(65.00%)以及A等位基因的分布频率(75.84%)明显高于稳定斑块组(30.00%和56.66%)(χ2=24.56、χ2=8.22,P<0.05)。结论 MMP-1基因-519A/G多态性与脑梗死的发病和颈动脉粥样硬化程度相关,AA基因型和A等位基因可能会降低颈动脉粥样硬化斑块的稳定性,增加脑梗死的发病风险。     

凝血酶活化纤维蛋白溶解抑制物基因编码区C1040T及G753A的多态性与脑梗死的相关性

目的探讨中国汉族脑梗死患者凝血酶活化纤维蛋白溶解抑制物(TAFI)基因编码区C1040T及G753A单核苷酸多态性与脑梗死的相关性。方法回顾性分析采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,检测130例脑梗死患者和118名同期行正常体检者(对照组)的TAFI基因编码区C1040T及G753A的多态性。结果脑梗死组TAFI基因G753A多态性的GG型频率41.5%(54例),A等位基因携带者频率58.5%(76例);而对照组分别为GG型频率44.9%(53例),A等位基因携带者频率55.1%(65例)。TAFI基因G753A多态性的GG型及A等位基因携带者两组频率差异无统计学意义(χ2=0.288,P=0.592)。在脑梗死组,C1040T多态性的CC型占50.0%(65例),T等位基因携带者占50.0%(65例);而对照组分别为CC型51.7%(61例),T等位基因携带者占48.3%(57例)。C1040T多态性的CC型及T等位基因携带者两组差异无统计学意义(χ2=0.071,P=0.790)。多因素Logistic回归分析显示,TAFI基因编码区G753A单核苷酸多态性(GA或AA基因型)及C1040T单核苷酸多态性(CT或TT基因型)不是脑梗死发病的独立危险因素。结论 TAFI基因编码区C1040T及G753A的多态性与脑梗死之间无显著相关性,不是脑梗死发病的独立危险因素。     

内皮细胞固有型一氧化氮合酶 4 b/a多态性与终末期慢性肾衰竭的关联性研究

目的对天津地区汉族人内皮细胞固有型一氧化氮合酶基因内含子4的插入/缺失多态性(ecNOS 4 b/a)与终末期慢性肾衰竭 (ESCRF)的关联性进行研究.方法应用PCR-小卫星DNA多态性分析技术,对67例ESCRF患者(观察组)和70例健康人(对照组)的ecNOS 4 b/a基因型分布进行检测.结果 (1) 观察组和对照组的三种基因型bb、ba和aa频率分别为79.1%、19.4%、1.5%、91.4%、8.6% 和0% .(2)两组等位基因分布差异有显著性(χ2=4.617,P＜0.05).观察组比对照组a/(a+b)OR值(95%可信区间)为2.64(1.09, 6.43)(Z=2.14,P＜0.05).(3)有a基因的ESCRF患者平均年龄(47.43±11.63)岁,无a基因的ESCRF患者平均年龄(58.08±13.68)岁,二者比较差异有显著性(t=2.664,P＜0.01).(4) 天津汉族正常人群a基因频率(4.3%)低于日本人(10.1%)(χ2=4.898,P=0.027).结论天津地区汉族健康人群a等位基因频率低于日本人和澳洲白种人;ecNOS 4 b/a多态性与天津地区汉族人ESCRF有关联性,a基因与ESCRF危险性增加呈正相关.     

脑梗死患者血小板膜糖蛋白Ⅰ bα基因HPA-2、Kozak位点多态性分析

目的观察脑梗死患者血小板膜糖蛋白（GP）Ⅰbα基因HPA-2、Kozak位点多态性,探讨其与脑梗死的关系。方法选取316例经CT或MR I证实的脑梗塞患者（观察组）及209例无脑血管疾病者（对照组）,采用聚合酶链反应—限制性片断长度多态性（PCR-RFLP）法检测GPⅠbα基因HPA-2、Kozak位点多态性,用流式细胞术（FCM）测定血小板活化标志物CD62P、CD63的表达。结果观察组血小板GPⅠbα基因HPA-2位点基因型分布及等位基因频率与对照组相比P均〉0.05;观察组Kozak位点观察组基因型为CC者20例、CT者113例、TT者169例,等位基因C频率为25.33%、T频率为74.67%;对照组分别为6、28、162例和10.20%、89.80%。两组基因型分布和等位基因C、T频率相比,P均〈0.05。观察组表达等位基因C者（基因型CC＋CT）CD62P表达量为20.94±7.35、CD63表达量为28.62±9.47,不表达等位基因C者（基因型为TT）分别为13.36±6.27、20.27±5.45,二者CD62P、CD63表达量相比P均〈0.05。结论 脑梗死患者血小板GPⅠbα基因Kozak位点存在多态性,HPA-2位点未发现多态性。GPⅠbα基因Kozak位点多态性可能通过影响血小板的活化参与脑梗死的发生过程。     

山东汉族人群血小板特异性抗原基因的多态性研究

目的 研究山东汉族人群血小板特异性抗原（HPA）1～5和15系统基因多态性分布特点。方法 采用PCR-序列特异性引物（PCR-SSP）技术对234例无血缘关系，汉族，血小板志愿捐献者进行HPA-1～5和HPA-15系统基因分型，计算等位基因频率、基因型频率并与其他种族、地区人群相关资料比较。结果 等位基因频率由高到低依次为HPA-4a、-1a、-5a、-2a、-3a、-15a、-15b、-3b、-2b、-5b、-1b，未检到HPA-4b。基因型频率由高到低依次为HPA-4a4a、-1a1a、-5a5a、-2a2a、-15a15b、-3a3b、-3a3a、-15a15a、-363b、-15b15b、-2a2b、-5a5b、-1a1b，未检到HPA-1b1b、-5b5b和HPA-2b2b。与报道的东方人群比较，有显著性差异的系统有HPA-2、-3,-5（P〈0．005）；与西方人群比较，HPA-1、-2、-3、-5系统有显著性差异（P〈0．005），HPA-4无种族间显著性差异。HPA-15系统的基因分布与越南、德国、奥地利人近似，而与印地安人有较大差异（P〈0．005）。结论 HPA存在明显的种族和地域性差异。山东汉族人发生HPA同种免疫的可能风险抗原基因主要有HPA-3a，-3b，-2b，-5b，-1b，其次为HPA-15a、-15b。     

Three layer functional model and energy exchange concept of aging process

Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.     

Unusual responses of nocturnal pineal melatonin synthesis and secretion to swimming: Attempts to define mechanisms

Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm²) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.     

Melatonin and photoperiodic time measurement: Seasonal breeding in the sheep

Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Serological survey of HIV and syphilis in pregnant women in Madagascar

Objectives Peripartal transmission of human immunodeficiency virus (HIV) and Treponema pallidum, the causative agent of syphilis, leads to severe consequences for newborns. Preventive measures require awareness of the maternal infection. Although HIV and syphilis testing in Madagascar could be theoretically carried out within the framework of the national pregnancy follow‐up scheme, the required test kits are rarely available at peripheral health centres. In this study, we screened blood samples of pregnant Madagascan women for HIV and syphilis seroprevalence to estimate the demand for systemic screening in pregnancy. Methods Retrospective anonymous serological analysis for HIV and syphilis was performed in plasma samples from 1232 pregnant women that were taken between May and July 2010 in Ambositra, Ifanadiana, Manakara, Mananjary, Moramanga and Tsiroanomandidy (Madagascar) during pregnancy follow‐up. Screening was based on Treponema pallidum haemagglutination tests for syphilis and rapid tests for HIV, with confirmation of positive screening results on line assays. Results Out of 1232 pregnant women, none were seropositive for HIV and 37 (3%) were seropositive for Treponema pallidum. Conclusions Our findings are in line with previous studies that describe considerable syphilis prevalence in the rural Madagascan population. The results suggest a need for screening to prevent peripartal Treponema pallidum transmission, while HIV is still rare. If they are known, Treponema pallidum infections can be easily, safely and inexpensively treated even in pregnancy to reduce the risk of transmission.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.