肝癌细胞株中肿瘤特异性抗原MAGE、GAGE、BAGE基因表达的研究

目的 研究MAGE、GAGE、BAGE基因在肝癌细胞株中的表达情况,评价这些肿瘤特异性抗原作为肿瘤分子标记以及肿瘤免疫治疗特异性靶位的可能性。 方法 用RT-PCR检测国内建株的肝癌细胞株SMMC-7721、QQY-7701、BEL-7402中MAGE-1、MAGE-3、GAGE1-8、GAGE1-2和BAGE基因mRNA的表达,以GAPDH基因作为检测内对照,并与非肿瘤肝穿组织比较。 结果 肝癌细胞株SMMC-7721表达MAGE-1和BAGE基因;QQY-7701表达MAGE-3和BAGE基因;BEL-7402表达MAGE-1和GAGE1-2基因;3株肝癌细胞至少表达其中一个基因。肝硬化病人肝穿刺组织中MAGE、GAGE、BAGE基因表达均为阴性。 结论 MAGE、GAGE、B A G E肿瘤特异性抗原可以作为肝癌早期诊断的分子标记,并具有作为肝癌免疫治疗特异性靶位的潜在价值。     

大肠癌组织中癌相关基因的表达及其启动子低甲基化的实验研究

大肠癌是最常见的消化系恶性肿瘤之一，近年来其发病率有逐年上升趋势。大肠癌的治疗关键在于早期诊断，而目前惟一的根治方法是早期切除肿瘤。癌相关基因（CAGE）是2002年发现的与癌症有关的基因，其在多种癌组织及肿瘤细胞系中呈高表达，在除睾丸组织外的正常组织中均无表达。现回顾性分析大肠癌、大肠腺瘤及正常黏膜组织中CAGE的表达及其启动子低甲基化情况。     

癌-睾丸抗原NY-ESO-1在视网膜母细胞瘤中的表达

目的了解视网膜母细胞瘤(RB)组织癌-睾丸抗原(CTA)基因NY-ESO-1的表达,探讨这些基因成为RB特异性免疫治疗靶抗原的可能性。方法选15例人RB组织和12例非肿瘤病变视网膜组织及22例正常眼部组织(对照组),采用逆转录-聚合酶链反应(RT-PCR)技术研究上述组织NY-ESO-1的mRNA表达情况,并根据肿瘤病理分级、肿瘤大小、临床分期等临床指标进行统计学分析。结果 15例RB组织中,NY-ESO-1的表达率分别为40%(6/15);NY-ESO-1基因在12例非肿瘤病变视网膜组织及22例正常眼部组织中均不表达。结论 NY-ESO-1基因在RB组织中有一定频率的表达,NY-ESO-1的表达率与病人的性别、年龄和临床分期,肿瘤大小及病理分级均无相关性(P0.05)。NY-ESO-1有希望成为RB特异性免疫治疗的靶抗原。     

GAGE-7自身抗体在食管鳞状细胞癌中的表达

目的:检测癌-睾丸抗原GAGE-7的自身抗体在食管鳞状细胞癌患者和正常人血清中表达的差异,评价GAGE家族作为肿瘤诊断标志物的可行性.方法:在原核细菌中表达并纯化重组GAGE-7蛋白.采用ELISA检测GAGE-7自身抗体在105份食管鳞状细胞癌患者血清和86份正常人血清中的表达水平.同时检测各血清中总IgG的表达量,计算GAGE-7自身抗体在总IgG中所占比例的差异.结果:血清中GAGE-7抗体在食管鳞状细胞癌中的含量与正常组比较差异显著(P<0.001),而GAGE-7抗体与总IgG比例,患者组明显高于正常组,逻辑回归模型分析c值为0.717,敏感性和特异性分别达到0.72和0.62,且差异显著(P<0.0001).结论:癌-睾丸抗原GAGE-7的自身抗体在食管鳞状细胞癌血清中有较高表达,且与正常组比较有显著差异,其表达特性可被看做是较为理想的癌症检测标志物.     

肝细胞癌中耐药基因的表达及意义

目的通过对肝细胞癌（HCC）中3种耐药基因MDR1、MRP、GST-π的检测,探讨肝细胞癌中耐药基因的表达特点及临床意义。方法应用逆转录-聚合酶链反应（RT-PCR）技术检测3种耐药基因在51例肝细胞癌组织和10例正常肝组织中的表达。结果 （1）MDR1、MRP、GST-π在肝细胞癌中的表达分别为0.55±0.27、0.62±0.29、0.64±0.31,正常肝组织中的表达分别为0.23±0.10、0.25±0.07、0.26±0.12,耐药基因在肝细胞癌中的表达高于正常肝组织,差异具有统计学意义（P〈0.05）;（2）耐药基因的表达与肿瘤Edmondson分级呈正相关（P〈0.05）;（3）MRP与GST-π的表达相关。结论肝细胞癌中存在有原发性耐药的现象,且多种机制并存。MDR1、MRP、GST-π在肝细胞癌中有较高的表达。联合检测对制定科学有效的治疗方案有一定价值。     

肝细胞癌组织中的miRNA-21基因表达变化

目的检测肝细胞癌组织中miRNA-21基因、RECK蛋白的表达变化。方法取60例肝细胞癌患者的肿瘤组织标本（观察组）和癌旁组织标本（对照组）,用PAGE/Northern blot分析法检测其中的miRNA-21基因表达。结果观察组miRNA-21基因表达表达量为19.3±3.39,对照组为2.35±1.63,两组相比P〈0.05。结论肝细胞癌组织中miRNA-21基因表达上调。     

DLC-1基因在肝细胞癌组织中表达的研究

目的 研究肝癌缺失基因-1（DLC-1）在人原发性肝癌组织中的表达及与肝细胞癌侵袭转移的关系。方法收集51例复旦大学附属中山医院外科手术切除的肝细胞癌及癌旁正常组织标本，用荧光定量RT-PCR方法分析组织中DLC-1基因的表达。结果在51对配对的肝细胞癌与癌旁无瘤肝组织中，前者DLC-1基因表达水平明显低于后者（P〈0．01），且高侵袭性肝细胞癌组明显低于低侵袭性肝细胞癌组（P〈0．05）。结论DLC-1基因在HCC组织中呈低表达，其表达水平与其侵袭性高低相关。     

DAC对卵巢癌细胞株增殖及其MAGE、BAGE、GAGE基因表达的影响

目的探讨5-杂氮脱氧胞嘧啶（DAC）对卵巢癌细胞株SKOV3、A2780、COC1增殖及其肿瘤特异性抗原MAGE、BAGE、GAGE基因表达的影响。方法将对数生长期的卵巢癌细胞株与DAC共同培养，观察细胞生长指数，利用流式细胞术检测细胞周期，RT-PCR技术检测卵巢癌细胞株在DAC作用前后MAGE、BAGE、GAGE基因的表达。结果DAC能使大多数巢癌细胞株阻滞在G0期，并使其MAGE、BAGE、GAGE基因表达增加。结论DAC能抑制卵巢癌细胞株的增殖，提高其肿瘤特异性抗原基因的表达。     

胃癌组织及血清中放免单克隆抗体癌相关抗原检测的研究

胃癌组织及血清中放免单克隆抗体癌相关抗原检测的研究徐萍，周铭伟，黄重发，袁新（解放军４５４医院）胃癌是我国最常见的恶性肿瘤之一，寻找具有诊断价值的肿瘤相关抗原一直是人们所关注的问题。我们应用新的放免测定盒检测胃癌组织及血清中单克隆抗体癌相关抗原（ＭＧ...     

泛癌组织中T-box转录因子19基因水平与患者预后和肿瘤免疫微环境的相关性

目的 基于数据库数据分析泛癌组织中T-box转录因子19(TBX19)表达水平与患者预后和肿瘤免疫微环境（免疫细胞浸润、免疫检查点基因、肿瘤突变负荷和微卫星不稳定性）之间的关系。方法 从癌症基因组图谱（TCGA）数据库中获取泛癌组织及相应正常组织中TBX19表达水平，比较不同WHO分级和TNM分期泛癌组织中TBX19表达水平，使用Cox回归分析TBX19表达水平与肿瘤患者预后的关系。基于肿瘤免疫估算数据库（TIMER2）数据，采用Pearson法分析TBX19表达水平与免疫细胞浸润丰度的相关性；在Sanger Box网站中利用TCGA数据库采用Pearson方法分析TBX19表达水平与免疫评分、肿瘤突变负荷和微卫星不稳定性的相关性；基于临床生信之家数据库数据，采用Spearman法分析TBX19表达水平与免疫检查点基因的相关性。结果 在膀胱尿路上皮癌、胆管癌、结肠癌（COAD）、结直肠癌（COADREAD）、食管癌、头颈鳞状细胞癌、肾透明细胞癌、肾乳头状细胞癌、混合肾癌、肝细胞癌（LIHC）、肺腺癌、肺鳞癌、嗜铬细胞瘤和副神经节瘤、直肠腺癌、胃癌、胃和食管癌癌组织中TBX19高表达（P...     

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.     

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.     

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.