流式细胞术分析脑益嗪对成纤细胞的调控作用

用流式细胞术测定不同剂量的脑益嗪（ＣＩＮ）对培养中３Ｔ６成纤维细胞的影响，包括检测细胞生长周期，ＤＮＡ含量，分裂增值指数及Ｇ１期和Ｇ２期蛋白质含量的变化。发现用脑益嗪后Ｇ１期细胞数增加，而Ｓ期细胞数减少，但无明显的量效关系，ＤＮＡ吟及分裂增殖指数下降；Ｇ１期及Ｇ２期蛋白质含量明显高于对照组，结果表明，脑益嗪可阻滞Ｇ１－Ｓ进程，使细胞堆积于Ｇ１期，减少ＤＮＡ含量及细胞分裂，增加蛋白质含量，使３Ｔ６细     

钙拮抗剂对3T6成纤维细胞的影响

采用流式细胞技术（ＦＣＭ）观察不同剂量的四种钙拮抗剂（Ｃａ－Ａ）汉防已甲素（Ｔｅｔ）、硝苯啶（Ｎｉｆ）、维拉帕咪（Ｖｅｒ）及脑益嗪（Ｃｉｎ）对３Ｔ６成纤维细胞生长增殖的影响，结果表明它们不仅可阻断Ｇ１→Ｓ期的进程，而且抑制Ｓ期ＤＮＡ的含量，但却增加Ｇ１和Ｇ２期蛋白质的含量，使细胞呈现不平衡生长，最终死亡。其中Ｔｅｔ的作用最为显著，呈一定的量效关系，此作用与阻断钙离子内流无关。提示钙拮抗剂对３Ｔ６成纤维细胞生长增殖的抑制可能是其抗肝纤维化的机制之一。     

钙离子拮抗剂等对成纤维细胞增殖的影响

目的研究硝苯啶（Ｎｉｆ）、脑益嗪（Ｃｉｎ）、Ａ２３１８７和氯丙嗪（ＣＰＺ）对３Ｔ６成纤维细胞增殖的影响．方法以含１０％小牛血清的ＲＰＭＩ１６４０培养基培育３Ｔ６细胞，分别加入３种浓度的Ｃｉｎ，Ｎｉｆ，Ａ２３１８７和ＣＰＺ，处理２４ｈ后用图像分析仪和流式细胞仪检测３Ｔ６细胞的ＤＮＡ，ＲＮＡ和总蛋白以及细胞周期．结果Ｎｉｆ剂量依赖性地延迟３Ｔ６细胞Ｇ２期向Ｍ期的进展；Ｃｉｎ，Ａ２３１８７和ＣＰＺ分别呈剂量依赖性地抑制３Ｔ６细胞Ｇ１期向Ｓ期的进程；Ｎｉｆ，Ａ２３１８７和ＣＰＺ促进细胞的ＲＮＡ合成，Ｃｉｎ抑制细胞ＤＮＡ及ＲＮＡ的合成；Ａ２３１８７和ＣＰＺ（１０μｇ）抑制细胞蛋白质的合成，Ｃｉｎ（３０μｇ）及ＣＰＺ（５μｇ）促进细胞蛋白质合成．结论Ｎｉｆ等通过对细胞周期进程的延迟及引起细胞不平衡生长，抑制３Ｔ６成纤维细胞的增殖．     

氟对体外骨器官培养长骨细胞周期和细胞凋亡的影响

为了探讨氟对体外骨器官培养长骨细胞周期分布和细胞凋亡的影响，我们采用体外骨器官培养的方法，流式细胞术检测长骨细胞周期分布、ＤＮＡ含量和细胞凋亡。结果发现１．染氟２．５￣５．０μｇ／ｍＬ，对体外培养长骨细胞ＤＮＡ含量和细胞周期分布影响不大。染氟１０．０μｇ／ｍＬ．Ｓ期细胞数增加，但Ｇｏ／Ｇ１期和Ｇ２／Ｍ期细胞无明显改变。染氟２０．０μｇ／ｍＬ，Ｓ期细胞数增加，Ｇ２／Ｍ期细胞数减少，但Ｇ０／Ｇ１期细胞     

汉防己甲素及维拉帕米对成纤维细胞生长增殖的影响

目的探讨汉防己甲素（Ｔｅｔ）及维拉帕米（Ｖｅｒ）抗肝纤维化的机制．方法采用细胞培养、流式细胞术和图像分析法检测Ｔｅｔ及Ｖｅｒ对３Ｔ６成纤维细胞增殖的影响．结果不同浓度的Ｔｅｔ及Ｖｅｒ可使３Ｔ６细胞Ｇ１期或Ｇ２＋Ｍ期细胞显著增多（Ｐ＜００１）；Ｔｅｔ（１５，２０μｇ）使细胞ＲＮＡ含量显著下降（Ｐ＜００１）；Ｖｅｒ（１０，２０μｇ）使细胞ＤＮＡ含量显著降低（Ｐ＜００１）；Ｔｅｔ（１５，２０μｇ）和Ｖｅｒ（５，１０μｇ）均使细胞蛋白质含量显著增加（Ｐ＜００１）．结论Ｔｅｔ及Ｖｅｒ对成纤维细胞的增殖具有抑制作用，可能是抗肝纤维化的机制之一     

亚硝酸盐对培养心肌细胞的直接作用

应用亚硝酸钠作用于培养的心肌细胞，观察到心肌细胞发生了超微形态结构改变；细胞内α－羟丁酸脱氢酶漏出增加（Ｐ＜０．０５～０．０１），３Ｈ—ＴｄＲ掺入变化呈双向性：小剂量组（０．００１～０．１×１０－６ｍｏｌ／Ｌ）呈现促进作用（Ｐ＜０．０５～０．０１）；大剂量组（１．０×１０－６ｍｏｌ／Ｌ）则表现为抑制效应（Ｐ＜０．０５）。细胞周期分析结果：小剂量组（０．０１×１０－６ｍｏｌ／Ｌ）Ｇ０Ｇ１期细胞减少４．１３％，Ｓ期增长３．９５％，Ｇ２＋Ｍ期增长０．３３％，与对照组比较，无显著性差异（Ｐ＞０．０５）。大剂量组（１、０×１０－６ｍｏｌ／Ｌ）Ｇ０Ｇ１期细胞增加９．１４％，Ｓ期减少１２．４６％，Ｇ２＋Ｍ期增加４．０６％，其中Ｓ期的改变达到显著差异（Ｐ＜０．０５）；细胞及培养液中脂质过氧化物含量增加（Ｐ＜０．０１）。表明亚硝酸盐能引起培养心肌细胞的直接损伤，其机理与脂质过氧化作用有关。     

空腹血胰岛素、葡萄糖比值作为β细胞功能指数的可能性

目的探讨空腹血浆胰岛素、葡萄糖比值（ＦＩＮＳ／ＦＰＧ）作为群体研究胰岛β细胞功能指数的可能性，并与文献中其他指数比较。方法在糖耐量正常者（ＮＧＴ）１８１例，糖耐量低减者（ＩＧＴ）２８１例测定空腹及服葡萄糖７５克后１小时、２小时静脉血浆葡萄糖及胰岛素。按服糖后２小时血糖水平依次递增０．５５６ｍｍｏｌ／Ｌ，再将ＩＧＴ分为血糖从低到高６个亚组，以ｔ检验及方差检验判定ＦＩＮＳ／ＦＰＧ及文献中常用五种胰岛β细胞功能指数分辨ＮＧＴ与ＩＧＴ，及ＩＧＴ血糖水平不同亚组间β细胞功能差别的能力。结果①ＯＧＴＴ１小时血浆胰岛素、葡萄糖比值（ＩＮＳ１／ＰＧ１），ＯＧＴＴ胰岛素、血糖增值比值（△ＩＮＳ／△ＰＧ）以及Ｈｏｍａｍｏｄｅｌ中的２０×ＦＩＮＳ／（ＦＰＧ３．５）都能分辨从ＮＧＴ、ＩＧＴ的β细胞功能变化，却无一能辨认ＩＧＴ血糖水平不同亚群的β细胞功能差别。②调整胰岛素敏感性的影响后，它们则具有了各种不同程度的分辨上述人群β细胞功能变化趋势的能力。结论①使用上述β细胞功能指数必须调整胰岛素敏感性的影响；②ＦＩＮＳ／ＦＰＧ可在流行病研究中评估β细胞功能，其分辨能力不逊于Ｈｏｍａｍｏｄｅｌ中的２０×ＦＩＮＳ（ＦＰＧ３．５）。     

平滑肌细胞源性生长因子的部分纯化

本文用超滤和肝素亲和层析法对培养的兔主动脉平滑肌细胞的无血清条件培养基进行纯化。用３Ｈ－ＴｄＲ掺入细胞ＤＮＡ法检测层析所得各组分对ＮＩＨ３Ｔ３纤维母细胞的致有丝分裂活性。用ＮａＤｏｄＳＯ４－聚丙烯酰胺凝胶电泳及等电聚焦电泳检测其分子量及等电点。结果表明，该条件培养基经肝素亲和层析，被１．０～１．６ｍｏｌ·Ｌ＇ＮａＣｌ洗脱下来的蛋白质成分对３Ｔ３细胞有致有丝分裂活性，其分子量范围为２３．０～２６．９ｋＤａ，等电点约为４．６。这些生物化学特性不同于ＰＤＧＦ、ＩＧＦ及ＦＧＦ，提示这种致有丝分裂蛋白质可能是一种独立的生长因子。     

DNA及增殖细胞核抗原含量与急性白血病临床及预后关系的研究

目的探讨急性白血病细胞ＤＮＡ及增殖细胞核抗原（ＰＣＮＡ）含量的临床意义和预后。方法用流式细胞仪（ＦＣＭ）对５６例急性髓细胞性白血病（ＡＭＬ），检测其骨髓单个核细胞ＤＮＡ含量及ＰＣＮＡ阳性率。结果白血病细胞周期结果显示，初治组Ｇ０／Ｇ１期百分率８７．４％±１１．３％显著高于完全缓解（ＣＲ）组４５．１％±１０．２％及对照组４８．４％±６．９％（Ｐ＜０．０５），同时初治组Ｓ期为８．９％±７．４％及Ｓ＋Ｇ２／Ｍ１５．７％±１０．１％低于对照组及ＣＲ组；亚二倍体组ＣＲ率５６．３％低于整二倍体９６．４％及超二倍体组９１．７％，ＰＣＮＡ阳性率分别与ＣＲ率、生存率呈负相关；ＰＣＮＡ阳性率与非整倍体ＤＮＡ检出率两者有显著相关性，呈正相关。初治组之间ｒ＝０．６４２，Ｐ＜０．０５，复发组之间ｒ＝０．５８３，Ｐ＜０．０５。结论将急性白血病患者ＰＣＮＡ阳性率与ＤＮＡ的含量相结合，才能正确全面评估急性白血病预后     

弓形虫病诊断方法的对比研究

应用５种不同方法测定了６例弓形虫病人不同病期的血清标木。用ＩＨＡＴ和ＩＦＡＴ测定抗弓形虫总免疫球蛋白，ＩＨＡＴ在发病后第３周的２份血清标本中有１份已呈阳性，１４～１７周达高峰。滴度升高非常明显；ＩＦＡＴ第３周的标本都呈阳性，１７～２４周达高峰。用ＩｇＧ－ＥＬＩＳＡ和ＩｇＭ－ＥＬＩＳＡ，ＩｇＭ－ＩＦＡＴ分别测定抗弓形虫特异性抗体ＩｇＧ和ＩｇＭ，ＩｇＧ－ＥＬＩＳＡ从第６周呈阳性，随病情延长，滴度明显升高；ＩｇＭ－ＥＬＩＳＡ两份第３周的标本都呈阳性，至少第１７周仍保持较高水平，其中１人第２４周的标本仍呈阳性；但ＩｇＭ－ＩＦＡＴ仅２例（２／６）呈阳性且滴度不很高，在较短时间内就转阴。结果表明：病人的抗体类型相似；ＩｇＧ－ＥＬＩＳＡ，ＩｇＭ－ＥＬＩＳＡ和ＩＦＡＴ敏感特异，ＩＨＡＴ简便、快速；高滴度特异性ＩｇＧ易掩盖ＩｇＭ－ＩＦＡＴ；ＩｇＭ－ＥＬＩＳＡ可为早期诊断提供依据。     

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.     

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.