弓形虫表面抗原P30DNA疫苗免疫小鼠诱导细胞免疫应答的研究

目的 观察弓形虫表面抗原P30DNA疫苗诱导小鼠产生的细胞免疫应答。方法 大量制备重组真核表达质粒pBK-P30,用生理盐水（NS）稀释至1μg/μl,1次肌注免疫BALB／c小鼠,分别在免疫后5和10周,通过ConA刺激,MTT法测定免疫鼠脾脏T淋巴细胞转化率;使用间接免疫荧光法,用流式细胞仪对CD4^ 、CD8^ T细胞亚群进行测定。结果 ConA刺激小鼠脾T淋巴细胞发生增殖反应,pBK-P30免疫组与对两对照组及对照组之间的差异均无显著性（P＞0．05）。T细胞亚群CD4^ 、CD8^ 动态分析,CD4^ T细胞在各组增殖均不明显（P＞0．05）,但免疫组CD8^ 细胞数量升高,CD4^ /CD8^ 比率下降,与空质粒pBK-CMV对照组和NS对照组之间差异均有显著性（P＜0．05,P＜0．01）,pBK-CMV对照组与NS对照组之间差异也有显著性（P＜0．01）。免疫后10周,与NS对照组相比较,免疫组和pBK-CMV对照组CD8^ 细胞仍升高（P＜0．01）,但它们之间的差异无显著性（P＞0．05）。结论 弓形虫表面抗原P30DNA疫苗能诱导BALB／c小鼠产生一定的细胞免疫应答。     

系统性红斑狼疮T细胞活化及其与病情活动的相关性研究

目的探讨系统性红斑狼疮(SLE)患者T淋巴细胞的活化及多种活化的T细胞与疾病活动指标之间的相关性。方法SLE患者28例(其中活动期16例)和健康对照18名。应用流式细胞术,比较活动期和非活动期SLE患者外周血T细胞CD25、人类白细胞抗原(HLA)-DR以及CD69的表达差别。用直线相关分析判断活化T细胞与SLE疾病活动性评分(SLEDAI),24h尿蛋白定量,血清补体C3、C4的相关性。结果活动期SLE CD3~ CD69~ 和CD4~ CD25~ 细胞明显低于正常对照组(P＜0．05),CD3~ HLA-DR~ 、CD4~ HLA-DR~ 、CD8~ HLA-DR~ 细胞均明显高于正常对照组(P＜0．01),非活动期与正常对照组之间差异无统计学意义(P＞0．05)。28例SLE患者CD3~ CD69~ 细胞与SLEDAI评分、24h尿蛋白之间均呈显著负相关(r=-0．858,r=-0．756,P＜0．01),与CD4~ 细胞之间呈显著正相关(r=0．79,P＜0．01)。活动期SLE患者CD3~ CD69~ 细胞与C4、CD4~ 细胞之间均呈显著正相关(r=0．757,r=0．666,P＜0．05)。非活动期SLE患者CD3~ CD69~ 细胞与C3之间呈显著正相关,CD4~ HLA-DR~ 细胞与C4之间呈显著正相关,CD3~ HLA-DR~ 、CD8~ HLA-DR~ 细胞与CD4~ 细胞之间呈显著负相关(P＜0．05)。结论SLE患者活动期T淋巴细胞存在异常活化,SLE活动期CD3~ CD69~ 细胞数量减少,CD~3 CD69~ 细胞可以作为反映病情活动的指标。     

梅毒患者细胞免疫的研究

目的：探讨T淋巴细胞亚群、可溶性白介素2受体和肿瘤坏死因子α在梅毒发病机制中的作用。方法：应用流式细胞仪和双抗体夹心ELISA法对86例梅毒患者Ｔ淋巴细胞亚群（CD3^ 、CD4^ 、CD8^ ）、可溶性白介素2受体（sIL-2R)及肿瘤坏死因子α（TNF－α）水平进行检测，并对其相关性进行分析。结果：梅毒患者CD4^ 及CD4^ ／CD8^ 比值明显低于正常人对照组（P＜0．01），CD8^ 显著高于正常人对照组（P＜0．05），而CD3^ 与正常人对照组无显著性差异（P＞0．05）；活动期梅毒CD4^ /CD8^ 比值低于恢复期（P＜0．05）。活动期梅毒及非活动期梅毒患者血清sIL-2R水平均明显增高（P＜0．01），且活动期高于非活动期（P＜0．01）。活动期梅毒患者血清TNF－α水平高于非活动期及正常对照组（P＜0．01），且活动期高于非活动期（P＜0．01）。活动期梅毒患者CD4^ 、CD8^ 与sIL-2R和TNF－α水平呈正、负相关。结论：本研究提示梅毒患者存在细胞免疫功能抑制现象，且T淋巴细胞功能的紊乱和TNF－α及sIL-2R水平的改变与梅毒的发病有密切关系。     

CD3单抗和淋巴细胞功能相关抗原-1对狼疮肾炎外周血单个核细胞的共刺激作用

目的：探讨CD3单抗（mAb)和淋巴细胞功能相关抗原－1单抗（LFA－1mAb)对狼疮肾炎（LN）患者外周血单个核细胞（PBMC）的共刺激作用。方法：2例LN患者被分为活动期（n=14)和非活动期（n=15)，以12例健康献血员为对照，观察CD3mAb或（和）LFA－1mAb共刺激及阻断1－磷酯酰肌醇3－激酶（PI3－K）对体外培养96h后PBMC增生和IL－2产生的影响。PBMC提取采用Ficoll密度梯度离心法，细胞增生实验采用^3H-TdR掺入法，IL－2测定采用ELISA法。结果：PBMC培养96h后，自然生长的LN非活动期和活动期PBMC ^3H-TdR掺入量和IL－2合成与正常对照无明显差异（P值均＞0或0．05）；与自然生长的PBMC相比，CD3mAb单独处理增加了LN非活动期（P值均＜0．05）和活动期（P值均＜0．05）PBMC^3H-TdR掺入量和IL－2合成。而单独CD3mAb对正常对照PBMC^3H-TdR掺入量和IL－2合成没有影响（P值均＞0．05）。与CD3mAb单独处理组相比，CD3mAb和LFA－1mAb共刺激均增加了LN活动期（P值均＜0．05）和非活动期（P值均＜0．05）及正常对照（P值均＜0．05）PBMC ^3H-TdR掺入量和IL－2合成；与对照组相比，CD3mAb和LFA－1mAb共刺激后活动期（P值均＜0．01）和非活动期（P值均＜0．01）PBMC^3H-TdR掺入量和IL－2合成均增加，但活动期效应明显强于非活动期（P＜0．01）。PI3－K特异抑制剂Wortmannin(WT)的加入明显抑制了CD3mAb和LFA－1mAb共刺激诱导的PBMC增生和IL－2的分泌效应（P＜0．01，P＜0．01）。结论：CD3和FLA－1对狼疮肾炎PBMC具有共刺激作用，这种共刺激作用可能参与了狼疮肾炎T、B细胞的异常活化，阻断共刺激传递信号分子PI3－K则能抑制CD3和LFA－1介导的共刺激作用。     

~(125)Ⅰ-抗Tg-McAb与甲状腺癌细胞结合能力的影响因素研究

模仿体内环境及放射免疫显像临床应用中遇到的实际问题，观察Tg浓度、pH值及碘化钾（KI）、青霉素和地塞米松等药物对1251-抗Tg－McAb与高分化型甲状腺癌细胞结合能力的影响作用．结果显示，Tg的影响与其浓度密切相关，Tg浓度＜200ng／ml时，1251-抗Tg－McAb与高分化状腺甲癌细胞的结合率没有明显影响作用（P＞0．05）；当Tg浓度＞500ng／ml时，影响作用显著（P＜0．01）．偏酸环境可使抗Tg－McAb与高分化型甲状腺癌细胞的结合率稍低，但影响作用不大（P＞0．05），偏碱条件下，标记McAb活性损失增大，结合率显著降低（P＜0．01）．KI和青霉素加地塞米松对结合能力无任何影响．     

老年高血压病患者外周血T淋巴细胞亚群及红细胞免疫功能的临床研究

为探讨老年原发性高血压（OEH）患者外周血T淋巴细胞亚群及红细胞免疫功能的临床意义，对38例OEH患者的外周血T淋巴细胞亚群及红细胞免疫功能进行测定，并与对照组比较。结果表明：OEH患者CD3＋、CD4＋、CD8＋细胞百分率均低于对照组，CD4＋／DS8＋明显高于对照组（P＜0．01），其中以CD8＋细胞数量变化尤为明显，OEH患者红细胞C3b受体花环率（RBC－C3b）明显降低，而红细胞免疫复合物花环率（RBC-ICR）明显升高（P＜0．01）；CD8＋细胞数量与RBC－C3b呈正相关（T＝0．667P＜0．05），而与RBC－ICR呈负相关（r＝0．721P＜0．05）；RBC－C3b与RBC－ICR呈负相关（r＝－0．581P＜0．05）提示：OEH患者存在免疫功能紊乱，外周血T淋巴细胞亚群与红细胞免疫功能关系密切。     

妇科手术患者围术期外周血T细胞亚群的变化及其与血浆儿茶酚胺的相关性

妇科择期手术患者20例。采用连续硬膜外麻醉．术毕前10min硬膜外注入吗啡2mg。于术前及术后第1、3天晨抽取空腹静脉血，测定外周血T细胞亚群以及血浆儿茶酚胺（CA）。结果显示，术后第1、3天外周血CD3^＋及CD4^＋细胞百分率较术前明显降低（P均〈0．01），而CA较术前明显升高（P〈0．01）。CD3^＋细胞百分率与CA水平呈负相关，r=-0．772，P〈0．01；CD4^＋细胞百分率与CA无直线相关关系，r=0．350．P〉0．05。提示妇科手术患者围术期细胞免疫及内分泌状态发生变化．CD3^＋细胞的变化与CA水平变化有关。     

Runx3过表达对慢性乙型肝炎患者Th1、Th2型细胞因子表达的影响

目的：探讨 Runx3过表达对 CHB 患者 Th1、Th2型细胞因子表达水平的影响。方法重组慢病毒载体pGC-FU-Runx3与阴性对照慢病毒载体 pGC-FU 分别转染29例 CHB 患者外周血 CD4＋ T 细胞,收集培养3 d、5 d 和7 d的细胞培养上清液,应用 ELISA 检测 Th1型细胞因子 IFN-γ、IL-2和 Th2型细胞因子 IL-4、IL-10的表达水平。结果与pGC-FU 转染组比较,pGC-FU-Runx3转染组 Th1型细胞因子 IFN-γ的表达水平在3 d（P ＜0．05）、5 d（P ＜0．01）和7 d （P ＜0．01）时均明显升高;IL-2的表达水平在3 d 时差异无统计学意义（P ＞0．05）,但在5 d（P ＜0．05）和7 d（P ＜0．01）时均明显升高。与 pGC-FU 转染组比较,pGC-FU-Runx3转染组 Th2型细胞因子 IL-4的表达水平在3 d 时差异无统计学意义（P ＞0．05）,但在5 d（P ＜0．01）和7 d（P ＜0．05）时均明显降低;IL-10的表达水平在3 d 时差异无统计学意义（P ＞0．05）,但在5 d（P ＜0．05）和7 d（P ＜0．05）时均明显降低。与 pGC-FU 转染组比较,pGC-FU-Runx3转染组 IFN-γ／IL-4比值在3 d（P ＜0．01）、5 d（P ＜0．01）和7 d（P ＜0．01）时均明显增大。结论 Runx3过表达可以促进 CHB 患者 Th1型细胞因子的分泌,抑制 Th2型细胞因子的分泌,使其 Th1／Th2失平衡得到改善。     

白细胞介素-8受体A型和B型在强直性脊柱炎中的表达及其意义探讨

目的 检测白细胞介素-8受体A型（CXCR1）和白细胞介素-8受体B型（CXCR2）在强直性脊柱炎（AS）患者外周血中性粒细胞、CD14^＋单核细胞和CD3^＋T细胞上的表达水平，探讨其与AS疾病活动的相关性和可能涉及的AS炎症发病机制。方法 研究对象包括30例活动期AS患者，30例活动期类风湿关节炎（RA）患者和30名健康对照者，应用流式细胞术（FCM）检测CXCR1、CXCR2分别在AS患者、RA患者和健康对照者外周血中性粒细胞、CD14^＋单核细胞和CD3^＋T细胞上平均荧光强度（MFI）的表达水平，并和AS患者的临床BASFI、BASDAI、红细胞沉降率（ESR）、血清C反应蛋白（CRP）等指标进行相关分析。结果 CXCR1在AS患者组外周血CD3^＋T细胞上MFI表达水平（41±24）分别较RA患者组（18±10）和健康对照组（19±7）高（P均〈0．01）。CXCR2在AS患者组外周血CD14^＋单核细胞MFI表达水平（210±54）较健康对照组（300±52）低（P〈0．01），与RA患者组（191±53）比较差异无统计学意义（P〉0．05）；CXCR2在AS患者外周血CD14^＋单核细胞上MFI表达下降与患者毕氏疾病功能指数（BASFI）（r=-0．394，P=0．031）、毕氏AS疾病活动指数（BASDAI）（rs=-0．378，P=0．040）、ESR（rs=-0．465，P=0.010）、CRP（rs=-0．648．P=0．000）存在着负相关关系。结论 CXCR1和CXCR2分别在AS患者外周血CD3^＋T细胞和CD14^＋单核细胞表达异常，提示它们可能参与了AS的发病过程。检测AS患者外周血CD14^＋单核细胞的MFI表达水平可能是评价AS疾病活动性有价值的潜在的生物学标志之一。     

艾滋病患者抗病毒治疗最佳时机探讨

目的探讨艾滋病抗病毒治疗的最佳时机。方法分析116例艾滋病患者的抗病毒治疗效果,统计累计治疗时间及治疗1a后不同治疗时机与治疗方案对CD4^＋细胞增长率的影响。结果89例存活,17例死亡,3例失访,7例退出治疗。CD4^＋细胞〈100开始治疗者病死率高于CD4^＋细胞≥100开始治疗者（P〈0．05）。89例存活的艾滋病患者治疗后CD4^＋细胞平均增加329．92个／μl,增长2．65倍;CD4^＋细胞100～200开始治疗者CD4^＋细胞增长率明显高于CD4^＋细胞〉200开始治疗者（F=7．74,P〈0．05）。结论艾滋病患者抗病毒治疗的最佳时机为CD4^＋细胞100-200。     

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.