肝癌组织及血清中细胞间粘附分子-1表达的研究

目的研究细胞间粘附分子－１（ｉｎｔｅｒｃｅｌｕｌａｒａｄｈｅｓｉｏｎｍｏｌｅｃｕｌｅ－１，ＩＣＡＭ－１）作为判断肝癌发展程度及转移状态指标的可能性。方法以免疫组化的方法检测ＩＣＡＭ－１在肝癌组织和正常肝组织中的表达，观察了ＩＣＡＭ－１表达在细胞的定位。以点免疫印迹法测定不同患者血清和不同肝组织中ＩＣＡＭ－１的表达。分析ＩＣＡＭ－１表达与肿瘤生长转移状态、肿瘤特性的关系。结果肝癌细胞ＩＣＡＭ－１表达阳性（阳性率为８００％），主要分布在细胞膜，正常肝细胞则为阴性。肝癌患者血清可溶性细胞间粘附分子－１（ｓＩＣＡＭ－１）水平高于正常人（Ｐ＜００１）及肝良性肿瘤患者（Ｐ＜００５），肝癌伴转移者高于无转移者（Ｐ＜００５）。肝癌组织中ＩＣＡＭ－１含量明显高于癌旁组织（Ｐ＜００１）及正常肝组织（Ｐ＜００１），而与肿瘤大小及有无包膜无关（Ｐ＞００５）；转移组肝癌中ＩＣＡＭ－１的表达明显高于非转移组（Ｐ＜００５），两组癌旁组织中ＩＣＡＭ－１表达无差别（Ｐ＞００５）。结论血清及组织中ＩＣＡＭ－１的水平在一定程度上可以反映肝癌发展程度及转移状态，有可能作为预测肝癌转移复发的指标。     

原发性肝癌癌组织肿瘤抑制因子候选基因1表达与病理特征的关系

目的观察原发性肝癌癌组织肿瘤抑制因子候选基因1(TUSC1)表达与病理特征的关系。方法纳入2016年10月至2018年10月于我院收治的78例原发性肝癌患者为研究对象,进行回顾性分析。采用实时荧光定量聚合酶链反应,检测癌组织TUSC1基因mRNA表达,并采用Western blot法检测TUSC1蛋白表达。根据TUSC1 mRNA检测结果,将其分为TUSC1 mRNA表达组(55例)及缺失组(23例);按TUSC1蛋白检测结果,将其分为TUSC1蛋白表达组(45例)及缺失组(33例)。观察原发性肝癌患者癌组织及癌旁组织中TUSC1 mRNA及其蛋白表达水平,分析原发性肝癌病理特征与癌组织中TUSC1 mRNA及其蛋白表达水平的关系,并观察TUSC1 mRNA表达与TUSC1蛋白表达的关系。结果原发性肝癌癌组织中TUSC1 mRNA及其蛋白表达水平显著低于癌旁组织(P0.05)。TUSC1 mRNA及其蛋白表达与原发性肝癌患者性别、年龄、肿瘤部位、肿瘤直径、肿瘤类型、HBsAg均无明显关系(P0.05),但中晚期(TNM分期为Ⅲ、Ⅳ期)者TUSC1 mRNA及其蛋白阳性表达率显著高于早期(Ⅰ、Ⅱ期)者(P0.05),高、中分化者TUSC1 mRNA及其蛋白阳性表达率显著低于低、未分化者(P0.05),肝内转移者TUSC1 mRNA及其蛋白阳性表达率显著高于肝内无转移者(P0.05),伴门静脉癌栓者TUSC1 mRNA及其蛋白阳性表达率显著高于不伴门静脉癌栓者(P0.05)。原发性肝癌患者TUSC1 mRNA表达与TUSC1蛋白表达具有良好一致性(kappa为0.56)。结论原发性肝癌患者癌组织中TUSC1 mRNA及其蛋白表达水平下降,与患者TNM分期、分化程度、门静脉癌栓、肝内转移有关。     

癌细胞端粒酶活性、DNA含量和Ki-67表达与肝细胞肝癌转移和复发的关系

为探讨端粒酶活性、DNA倍体和Ki－67基因在肝细胞性肝癌（HCC）中的表达及其相互关系，以及对肝癌转移和预后的影响，用TRAP法检测了34例HCC及15例肝硬化（门脉高压）患者肝组织端粒酶活性；用流式细胞计检测DNA倍体和Ki-67阳性细胞。结果显示，34例HCC中29例端粒酶活性，15例肝硬化中2例阳性，HCC二倍体肿瘤23例，异倍体肿瘤11例；端粒酶阳性表达和异倍体肿瘤与HCC肝内转移和门静脉癌栓明显相关（P＜0．05），而与肿瘤大小、AFP水平、有无乙型肝炎（HBV感染）及分化程度无关；Ki－67抗原在HCC中呈现高表达,与肝硬化组织相比差异显著（P＜0．01），Ki－67表达和端粒酶活性与肝内转移等因素的关系极为相似。提示大多数HCC和个别肝硬化组织端粒酶活性呈现阳性；Ki－67在HCC中呈现高表达；端粒酶活性、Ki－67基因和DNA倍体可能在肿瘤发生、转移及预后中起重要作用，尤其前者可作为临床诊断、病情恶性程度及预后判断的标记物。     

肝癌组织中FAM96A的表达及其与临床病理特征、预后的关系

目的研究肝癌组织中96序列相似的家庭成员(FAM96)A的表达及其与临床病理特征、预后的关系。方法以2014年2月至2016年2月德阳市第二人民医院收治的肝癌患者112例,分别采用实时荧光定量聚合酶链式反应与Western印迹法检测肝癌组织与癌旁组织中FAM96A的mRNA及蛋白表达情况,分析肝癌组织中FAM96A蛋白表达情况与临床病理特征及预后的关系。结果肝癌组织中FAM96A的mRNA及蛋白相对表达量,均显著低于癌旁组织(P<0.05)。肿瘤直径<5 cm、无门静脉癌栓、肝外转移阴性、Child分级A级、TNM分期Ⅰ～Ⅱ期肝癌患者组织中FAM96A的蛋白相对表达量均显著高于肿瘤直径≥5 cm、有门静脉癌栓、肝外转移阳性、Child分级B级、TNM分期Ⅲ～Ⅳ期患者(均P<0.05)。肝癌组织中FAM96A蛋白低表达组术后1年复发率、术后2年复发率,均显著高于FAM96A蛋白高表达组(均P<0.05)。结论肝癌组织中FAM96A存在明显低表达,其表达水平的升高往往预示着预后良好。     

人肝细胞癌中整合蛋白α5β1表达的生物学意义

为了探讨整合蛋白α5β1在肝细胞癌中表达状况及其生物学意义，本文应用Northern，印迹杂交、免疫组化及图像分析技术对肝癌细胞株7721、人肝细胞癌(HCC)组织作整合蛋白α5β1表达分析。结果表明，整合蛋白α5、β1 mRNA在肝癌细胞中表达量明显低于正常肝细胞，且与肝癌的分化程度相关；肝癌浸润部位癌细胞整合蛋白α5β1表达减少；伴肝内转移组其原发灶内α5、β1表达水平较无肝内转移组明显下降，但其静脉内癌栓外周部癌细胞表达增强。结果提示，整合蛋白α5β1的表达对HCC分化、浸润、转移起重要作用。     

检测细胞间粘附分子-1对原发性肝癌的诊断意义

1资料与方法：患者血清标本-80℃保存,HCC44例,LC30例,健康献血员30例。组织标本：经病理证实的HCC标本35例,正常肝组织6例。smICAM－1检测用免疫组织化学给台多媒体彩色病理图文分析系统分析,ICAM－1用相对定量,即ICAM－1量＝（样品灰度－本底灰度）×样品面积。血清sICAM－1应用ELISA法检测。统计学分析用t检验及直线相关分析。2.sICAM－1测检的结果：（1）LC组（242.3±96.1）U／ml与HCC组（286.4±83．4）U／mlsICAM－1水平均明显高于正常对照组（90.6±397）U／ml（P＜0．01）HCC组sICAM－l水平高于LC组（…     

肝癌组织中HIF-1α与VEGF的表达及意义

目的探讨缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）在肝细胞肝癌（HCC）发生、发展中的作用。方法采用免疫组化法检测30份HCC组织（HCC组）和10份正常肝组织（对照组）标本中HIF-1α和VEGF蛋白表达。结果HCC组HIF．10r．阳性率为73．3％,对照组为0％（P〈0．05）。HCC有肝内转移者HIF-1α表达明显高于无肝内转移者（P〈0．05）。HCC组VEGF阳性率为60．0％,对照组为10．0％（P〈0．05）。HCC有肝内转移、癌栓形成者VEGF表达明显高于无肝内转移及癌栓形成者（P均〈0．05）。HCC中HIF-1α与VEGF的表达呈正相关（r=0．431,P〈0．05）。结论HIF-1α、VEGF在HCC中表达上调,且在HCC的发生、发展中起促进作用。     

老年肝癌患者血清TGF-β1、Smad4水平及临床意义

目的探讨老年肝癌患者血清转化生长因子(TGF)-β1、Smad4水平及临床意义。方法选择浙江省肿瘤医院2015年1月至2016年12月收治的老年原发性肝癌患者80例作为肝癌组,老年健康体检者80例作为对照组。采用酶联免疫吸附试验(ELISA)测定血清TGF-β1、Smad4水平。结果肝癌组血清TGF-β1、Smad4水平明显高于对照组(P0.05)。肝癌患者血清TGF-β1、Smad4水平与性别、肿瘤大小、是否合并肝硬化无关(P0.05),与TNM分期和门静脉癌栓有关(P0.05),TNM分期Ⅲ～Ⅳ期患者血清TGF-β1、Smad4水平明显高于Ⅰ～Ⅱ期患者,有门静脉癌栓患者血清TGF-β1、Smad4水平明显高于无门静脉癌栓患者(P0.05)。死亡肝癌患者血清TGF-β1、Smad4水平明显高于存活肝癌患者(P0.05)。肝癌患者血清TGF-β1水平与Smad4水平呈正相关(r=0.537,P=0.000)。结论老年肝癌患者血清TGF-β1、Smad4水平明显升高,TGF-β1、Smad4在老年肝癌的发病过程中及预后评价中具有重要作用。     

原发性肝癌患者外周血、肝组织中GGT mRNA-H表达及意义

目的探讨判断原发性肝细胞癌血行转移的方法。方法以RT-PCR法检测38例原发性肝癌（HCC）患者外周血γ-谷氨酰转移酶mRNA亚型（GGTmRNA-H）表达及其中18例患者肝组织GGTmRNA-H表达。结果外周血GGTmRNA．H表达阳性率为42．1％（16、38）。相关分析显示,GGTmRNA-H阳性率与肝癌临床分期、肝内转移、门静脉癌栓和远处转移密切相关。肝癌组织GGTmRNA-H表达的阳性率为93．8％。结论血循环中GGTmRNA-H表达可判断HCC有无血源性播散;对AFP阴性或低值HCC患者有补充诊断作用。     

sICAM-1和GPC-3 mRNA对肝癌诊断与转移监测的临床价值

目的:定量分析肝病患者血可溶性细胞间粘附分子-1(sICAM-1)浓度和外周血单核细胞磷脂酰肌醇蛋白多糖-3(GPC-3)mRNA,探讨对肝癌(HCC)诊断与预后的价值.方法:收集住院肝病患者外周血,分离单核细胞,制备总RNA,经逆转录合成GPC-3cDNA,以荧光定量PCR扩增;并以酶联免疫吸附法(ELISA)定量分析血ICAM-1水平.结果:肝病发展过程中血ICAM-1表达呈梯度增加,HCC患者血ICAM-1表达显著高于肝硬化(t=3.184,P=0.002)和慢性肝炎患者(t=3.962,P<0.001),与伴门脉癌栓(t=2.941,P=0.005)及肝外转移(t=3.282,P=0.002)明显相关,与患者年龄、性别、HBsAg阳性与否、AFP浓度及肿瘤大小间未见明显相关.GPC-3mRNA阳性仅见肝癌患者(70.9%);肝硬化、慢性肝炎患者及正常对照组中未检出(2=26.773,P<0.001).GPC-3mRNA阳性表达与HBsAg阳性(2=14.601,P<0.001)、肝癌TNM分期(2=17.732,P<0.001)、伴门脉癌栓及肝外转移(2=22.271,P<0.001)显著相关,与瘤体直径、数目、AFP浓度及分化程度未见明显相关;两者可互补诊断,提高诊断肝癌阳性率.结论:sICAM-1和GPC-3mRNA检测是肝癌诊断和转移监测的良好标志物,且对AFP阴性肝癌具有互补诊断价值.     

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.     

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.     

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.