加拿大法裔人群中与肥胖相关的高血压的全基因组连锁分析

原发性高血压是一种由多种机制共同作用的多基因疾病。由肥胖导致的病理改变是与其相关的一个信号通路。我们主要研究高血压的一个亚型——肥胖相关的高血压，以寻找其致病基因。我们入选了55个有两个患病同胞或者更多患者的家系，他们均来自于地理位置偏远的加拿大法裔人群，其中15个家系的肥胖发病率较高（≥70％）。我们在所有的高血压家系和有高肥胖发病率的高血压家系中利用多位点连锁分析方法进行全基因组扫描（GeneHunter 2．1；位点密度：10cm）。在所有55个家庭的扫描中，我们在1号染色体和11号染色体上分别发现了两个连锁值显著（LOD score=2．5）的位点D1S1597和D11S1999。我们对仅患有肥胖高血压的家系进行扫描，在1号染色体的同一区域内发现连锁值最显著（LOD score=3．1）的一个位点是D1S1597。然后我们又D1S1597周围的其他遗传标记进行基因分型，得到了一个连锁更加显著的位点D1S2672（LOD score=3．5）。与肥胖高血压相关的许多候选基因都位于这个区域，其中包括肿瘤坏死因子受体2（tumor necrosis factor receptor 2）和心房利钠肽基因（atrial natriuretic peptide genes）。这些结果表明，研究临床定义明确的高血压亚型可能有助于寻找、鉴定复杂性疾病的致病基因，例如本研究中利用肥胖相关的高血压亚型。     

常染色体隐性骨质硬化症易感基因定位

目的本研究对一个常染色体隐性遗传的骨质硬化症家系进行了研究,以确定本家系骨质硬化症的致病基因。方法采用双能X线吸收法(DXA)测定骨密度(BMD)。提取11个家系成员及180名正常对照者外周血DNA,用基因测序排除已知与骨质硬化症相关的11个基因的突变后,做全基因组扫描、精细基因定位及单倍型分析确定致病基因候选区。采用TRAP染色试剂盒行骨活检标本破骨细胞染色。结果该常染色体隐性骨质硬化症家系无已知与骨质硬化症相关的基因突变。全基因组扫描、精细基因定位及单倍型分析结果显示,染色体19q13.2～q13.3的D19S197～D19S545之间的8.36cm范围为致病基因候选区,连锁分析最大LOD值Zmax=2.907(θ=0时)。骨活检标本的TRAP染色结果显示,与性别和年龄匹配的正常人相比,骨质硬化患者髂嵴破骨细胞数量减少。结论该常染色体隐性骨质硬化症家系患者破骨细胞减少,19号染色体q13.2～q13.3有骨质硬化症易感基因存在,为尚未报道的常染色体隐性骨质硬化症特色类型。     

2个汉族肥厚型心肌病家系致病基因与TCAP基因的关系研究

目的:确定候选基因TCAP与两个汉族家系家族性肥厚型心肌病(HCM)之间的连锁关系.方法:在排除13个已知家族性HCM致病基因与这两个汉族家系家族性HCM的连锁关系基础上,选择TCAP基因作为这两个汉族HCM家系的候选致病基因,在其所在的染色体区域选取4个微卫星标记(Marker)进行单倍型连锁分析.结果:D17S1814、D17S838、D17Sac091178和D17S1818这4个微卫星标记在重组率θ=0时,家系1的LOD值在-2.689754～-0.645666范围内,家系2的LOD值在-1.396476～0.416726之间;在重组率θ=0.1时,两个家系中最大的LOD值仅为0.272605.结论:TCAP基因与这两个汉族家系的HCM无连锁关系,TCAP基因不是这两个家系的致病基因,提示这两个汉族家系的致病基因可能是全新的未知致病基因.     

预激综合征与染色体7q3连锁

目的：探寻预激综合征的致病基因。方法：应用基因分型的方法，以7q3上D7S505、D7S688和D7S483为候选位点，对3个家庭性预激综合征家系（共101例成员）进行了连锁分析。结果：家庭性预激综合征与上述3个位点均连锁，以D7S505 Lod值最高，重组率（θ）为0．1时Lod=6.4；D7S483和D7S688位点Lod值分别为2．5和5．3。结论：预激综合征的相关基因位于7q3上。     

家族性致心律失常性右室心肌病与微卫星遗传标志的连锁分析

目的 研究探索致心律失常性右室心肌病(ARVC)家系与3个微卫星遗传标志的连锁关系，进行ARVC的基因定位。方法 选择微卫星遗传标志：D2S152、D14S252和D10S1664，对121例背景人群和5个中国人ARVC家系(家系编号1～5)，用每个微卫星引物扩增家系和背景人群DNA的微卫星片段，在DNA手工测序电泳仪槽上进行恒功率变性聚丙烯酰胺凝胶电泳、银染、读取等位基因片段，在常染色体显性和隐性两种遗传模式下进行连锁分析。结果 (1)根据D2S152的连锁资料，在常染色体显性遗传模式下，1～5号家系的连锁值(Logarithm of odd，LOD值)分别为2．17、-0．59、-∞(负无穷大)、-(表示此遗传模式下不支持连锁分析)、-∞，均为0=0。在常染色体隐性遗传模式下，1～5号家系的LOD值分别为-∞、-∞、-∞、-∞、0．18。(2)根据D14S252的连锁资料，在常染色体显性遗传模式下，1～5号家系的LOD值分别为-、-、-∞、-、0。在常染色体隐性遗传模式下，1～5号家系的LOD值分别为-∞、-0．81、-∞、-∞、0．09。(3)根据D10S1664的连锁资料，在常染色体显性遗传模式下，1～5号家系的LOD值分别为-、-、0．54、-、0．60。在常染色体隐性遗传模式下，1～5号家系的LOD值分别为-、-∞、-∞、-∞、-∞。结论 (1)1号家系与D2S152的连锁值已达2．17，连锁与不连锁的可能性之比为150：1，提示在D2S152附近存在ARVC的可能致病基因；排除3号家系与D2S152的连锁关系；需要收集更多资料才能决定2、4和5号家系是否与此位点连锁。(2)排除3号家系与D14S252的连锁，需要收集更多资料才能决定1、2、4和5号家系是否与此位点连锁。(3)5个家系与D10S1664的连锁关系都需收集更多资料。     

中国汉族Graves病的一个主要易感位点定位于染色体5q31

目的　确定中国人Graves病 (GD)的染色体易感区。方法　采用 2 77个高度多态 (平均间距约为 13cM )的微卫星标记对 5 4个中国汉族GD多发家系的 3 2 2名个体做全基因组扫描研究。分别计算疾病在呈显性及隐性遗传的各外显率下 ,各微卫星的两点LOD值及多点LOD值 ,并计算多点非参数连锁(NPL)值。结果　参数法连锁分析显示染色体 5q3 1区的D5S43 6的两点LOD值与多点LOD值均最高 ,分别为 2 .8和 2 .3。为了进一步确定该区域在GD发生中的作用 ,在D5S43 6两侧又增加了 4个微卫星做基因分型和连锁分析 ,在D5S2 0 90处得到了最大两点LOD值 4.3 1和最大多点LOD值 4.12 (具有遗传异质性 ,α^ =0 .3 8)。多点非参数法连锁分析也显示D5S2 0 90的NPL值最高 ,为 2 .66(P =0 .0 0 1) ,同样支持该位点与GD相连锁。结论　GD的一个主要易感位点位于染色体 5 q3 1区 ;在GD中存在遗传异质性。     

6个长QT综合征家系的分子遗传学检测

目的对6个先天性长QT综合征(long QT syndrom e,LQTS)家系成员进行基因检测。方法运用位于KCNH2和SCN5A基因内和临近的短串联重复(short tandem repeat,STR)(D7S1824、D7S2493、D7S483、D3S1298、D3S1767、D3S3521)位点确定染色体单体型。6个家系的所有成员进行单倍型连锁分析,确定基因型。1个家系经直接测序确定其基因型。结果家系1的致病基因位于染色体LQT3位点,而家系2~6的致病基因位于LQT2位点,家系6经直接测序确定该家系的先证者为散发病例,突变基因为KCNH2。确诊LQTS患者22例,其中6例为无症状基因携带者,排除6例可疑患者。结论LQTS的遗传学研究检测不但能确定LQTS基因分型,而且可进行LQTS的症状前诊断。从而为临床的基因靶向治疗提供依据。     

12个水盐代谢、离子转运候选基因与高血压病连锁分析

目的研究12个水盐代谢、离子转运调节基因与高血压病（EH）的关系，以筛选EH易感基因。方法在每一候选基因的染色体区域附近，选择微卫星DNA多态性位点作为遗传标记，采用微卫星引物荧光标记-基因扫描及分型技术，对95个EH核心家系的477个成员进行微卫星位点与EH连锁分析。遗传统计采用Genehunter软件包中两点非参数连锁（NPL）分析、最大LOD值及传递不平衡检验（TDT）。结果NPL检验显示，D12S398的Z=2.08,P＜0.05；LOD值=1.26,P＜0.01；TDT分析χ2=9.00,P＜0.005。其余位点NPL分析及TDT的P值均>0.05,LOD值<1。结论采用三种不同的遗传统计方法提示D12S398位点与EH存在连锁，并且在D12S398附近有血管加压素1A受体基因,该区域值得进一步研究。     

肥厚型心肌病MYH7基因致病突变的产前遗传诊断研究

目的在致病突变明确的家系中通过产前遗传诊断来阻断肥厚型心肌病(hypertrophy cardiomyopathy,HCM)的代际传递,减少HCM患者数量。方法对分别携带MYH7基因Arg663Ser和Arg453His致病突变的2例HCM患者进行家系分析,通过Sanger测序检测致病突变位点,通过毛细管电泳进行短串联重复序列(short tandem repeats,STR)分型及及连锁分析。在17-20周行羊穿术采集羊水,羊水细胞提取基因组DNA,进行遗传检测。结果MYH7基因Arg663Ser和Arg453His突变在各自家系中与HCM家系共分离,为所在家系的致病突变。我们选择了杂合度高的6个位于MYH7基因附近的STR(D14S50、D14S283、D14S990、D14S972、D14S64和D14S264)进行分型检测,确认在第一个家系中D14S50的173bp长度等位基因和D14S990的151bp长度等位基因与Arg663Ser突变连锁。在第二个家系中,D14S50的169bp长度等位基因和D14S283的145bp长度等位基因与Arg453His突变连锁。羊水DNA的Sanger测序和STR分型均显示2例胚胎均未携带MYH7基因致病突变。新生儿脐带血的复检结果与产前诊断结果一致。结论产前遗传诊断能够在孕早期明确诊断胎儿是否携带家族HCM致病突变,为咨询者夫妇提供合理的遗传咨询依据,对阻断HCM在家系中遗传具有重要意义。     

一个常染色体显性遗传性垂体性尿崩症家系AVP-NPⅡ基因的连锁和突变分析

目的 寻找中国人常染色体显性遗传性垂体性尿崩症（autosomal dominant neumhypophyseal diabetesinsipidus，ADNDⅠ）的致病基因。方法 采集ADNDⅠ家系，抽提基因组DNA，对AVP-NPⅡ基因进行微卫星标记（STR）连锁分析和PCR直接测序。结果 连锁分析结果显示，AVP-NPⅡ基因位点与ADNDⅠ表型连锁，但PCR产物直接测序未发现任何碱基的改变。结论 推测ADNDⅠ存在遗传多样性，很可能有其他致病基因突变影响机体正常水代谢，导致ADNDⅠ的发生。     

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.     

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.     

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.     

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.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Language of CTO interventions – Focus on hardware

The knowledge of variety of chronic total occlusion (CTO) hardware and the ability to use them represents the key to success of any CTO interventions. However, the multiplicity of CTO hardware and their physical character and the terminology used by experts create confusion in the mind of an average interventional cardiologist, particularly a beginner in this field. This knowledge is available but is scattered. We aim to classify and compare the currently used devices based on their properties focusing on how physical character of each device can be utilized in a specific situation, thus clarifying and simplifying the technical discourse.