IL-12在抗伯氏疟原虫感染中的免疫佐剂作用

目的　探讨IL - 12在抗伯氏疟原虫感染中的免疫佐剂作用。方法　BALB/c小鼠分别皮下用抗原sAg、IL - 12、抗原sAg联合IL - 12或抗原sAg联合弗氏佐剂进行免疫 ,共免疫 3次 ,每次间隔 2wk ,末次免疫后 1wk以 5× 10 5个感染P berghei的RBC攻击感染 ,观察各组小鼠原虫血症变化及存活时间。攻击感染后 1wk收集各组小鼠血清 ,采用ELISA检测血清中IFN -γ、IL - 4、IL - 10及IgG水平。结果　抗原sAg联合IL - 12免疫可较其它各免疫组及未免疫对照组显著延长小鼠存活时间 ,且全部小鼠在原虫血症达 5 0 %时开始死亡 ;而单独抗原sAg、单独IL - 12或抗原sAg联合弗氏佐剂免疫与未免疫比较 ,对小鼠原虫血症及存活时间均无明显影响 ,且半数以上小鼠均在原虫血症小于 2 5 %时死亡。抗原联合IL - 12免疫组小鼠血清中IFN -γ及IgG水平较单独抗原sAg、单独IL - 12及未免疫组显著升高。 结论　结果表明 ,IL - 12与sAg抗原共同免疫小鼠可诱导对P berghei攻击感染的部分抵抗力 ,具有一定的免疫佐剂活性。     

香菇多糖对约氏疟原虫感染BALB/c小鼠Th1型应答的调节作用

目的探讨香菇多糖(Lentinan,Lent)对致死型约氏疟原虫(Plasmodium yoelii 17XL,Py17XL)感染BALB/c小鼠Th1型细胞免疫应答的调节效应。方法对Py17XL感染的BALB/c小鼠进行不同时间点的Lent预处理,动态观察用药后各组感染小鼠原虫血症水平和生存率;于感染后第0d、1d、3d和5d分别提取小鼠脾细胞,ELISA法检测脾细胞培养上清中IL-12、IFN-γ的分泌水平,Griess反应检测脾细胞培养上清中一氧化氮(NO)含量。结果与药物未处理组相比,感染前15d 1mg/kg Lent用药组显著降低感染小鼠的原虫血症水平,提高生存率;明显增强Th1型免疫应答中关键细胞因子IL-12、IFN-γ的分泌水平,并提高NO含量。结论Lent预处理能够有效激发Py17XL感染的BALB/c小鼠Th1型细胞免疫应答的建立,提示调控免疫应答对于致死型约氏疟原虫感染早期免疫防御的重要性。     

IL-12对伯氏疟原虫红内期感染小鼠Th1/Th2细胞因子表达水平的影响

目的观察白细胞介素12 (IL-12) 对伯氏疟原虫(P.b)红内期感染小鼠Th1/Th2细胞因子表达水平的影响,探讨IL-12介导Th1/Th2免疫偏移在抗P.b红内期感染中的免疫调节作用.方法 BALB/c小鼠接种5×105个感染P.b的红细胞,同时分别给予0.03 μg/d或0.15 μg/d IL-12处理,用ELISA方法检测P.b感染小鼠血清或脾淋巴细胞体外培养上清中细胞因子IFN-γ、IL-4表达水平的动态变化.结果 0.03 μg/d IL-12处理组小鼠接种感染后第7天取脾淋巴细胞体外经PHA或可溶性抗原sAg刺激培养产生的IFN-γ水平较感染对照组显著升高,IL-4水平显著下降,而0.15 μg/d IL-12处理组脾淋巴细胞产生的IFN-γ及IL-4水平均较感染对照组显著下降.在感染后第3天,0.03 μg/d或0.15 μg/d IL-12处理组小鼠血清中IFN-γ均已出现,第5天达高峰,但感染后第7天 0.15 μg/d IL-12处理组血清中IFN-γ迅速下降,甚至低于感染对照组及0.03 μg/d IL-12处理组,感染对照组直到感染后第7天血清中方可检测到IFN-γ水平.结论适当低剂量IL-12诱导CD4 Th1免疫应答,产生细胞因子IFN-γ,以诱导抗P.b红内期感染的免疫保护作用;而较大剂量IL-12抑制机体抗感染免疫,甚至可致免疫病理损害.     

致死型约氏疟原虫感染DBA/2小鼠保护性免疫机制的研究

目的 观察约氏疟原虫(P.yoelii) 17XL感染DBA/2小鼠的免疫应答机制及免疫效应的动态变化。方法 1×10⁶ P.yoelii 17XL感染的红细胞经腹腔接种DBA/2小鼠, ELISA测定血清白细胞介素-12 (IL-12)、干扰素-γ( IFN-γ)、IL-4和IL-10的水平以及特异性抗体IgG水平。Griess反应检测脾细胞培养上清中一氧化氮(NO)含量。检测小鼠原虫血症、单核细胞百分率,观测其吞噬疟原虫现象。结果 感染小鼠第9天原虫血症高达46.9%,多数小鼠于感染后第20天左右自愈。感染后第6至16天,外周血查见有吞噬作用的单核细胞。感染后第1天起, IL-12水平开始升高; IFN-γ于第6天达最高水平, IL-4和IL-10分别于第9天和第15天达最高水平。脾细胞培养上清NO含量,分别于第6天和第20天显著升高。血清中特异性抗体IgG水平呈增高趋势,至第70天达最高水平。结论 Th1细胞的有效活化对遏制原虫血症和最终清除疟原虫具有重要意义。约氏疟原虫感染早期,单核-巨噬细胞对原虫血症的遏制起到关键作用。     

约氏疟原虫感染不同小鼠免疫分子的应答差异

目的探讨在约氏疟原虫感染过程中不同宿主的免疫应答差异。方法以约氏疟原虫（致死型）感染DBA／2和BALB／c小鼠，计算红细胞感染率；收集感染前和感染后1、3、6、9、12、15、20d小鼠血清，并无菌取出脾脏，培养脾细胞。应用ELISA试剂盒检测小鼠血清中IFN-γ和IL-12水平，并通过Griess方法检测脾细胞培养上清中NO含量。结果DBA／2小鼠的原虫血症峰值水平明显低于BALB／c小鼠，并于感染后第20d左右自愈；BALB／c小鼠于原虫血症达到峰值水平后全部死亡；DBA／2小鼠的IFN-γ和IL-12水平于感染后1d即出现了有意义的升高并持续到第20d；BALB／c小鼠的IFN-γ和IL-12水平仅干感染后1d出现了有意义的升高；DBA／2小鼠NO的产生于感染后3d出现了有意义的升高，第6d达到峰值，而BALB／c小鼠的NO水平始终未见明显升高。结论DBA／2小鼠通过感染早期Th1细胞免疫应答的有效建立能够抑制原虫血症，IL-12是启动并维持Th1细胞免疫应答的关键性细胞因子。     

脂多糖对约氏疟原虫感染DBA/2小鼠感染早期的免疫调节作用

目的本文主要探讨脂多糖（LPS）在致死型约氏疟原虫（Plasmodium yoelii 17×L,Pyl7×L）感染早期的调节作用。方法 6～8w、雌性DBA/2小鼠,随机分为实验组和对照组,经腹腔接种1×106 Pyl7XL寄生的红细胞。实验组于感染后d2静脉给予LPS（25μg/只）。动态观察两组小鼠的原虫血症水平,小鼠脾脏树突亚群及其表面TLR4和活化T细胞的表达水平。结果 （1）两组小鼠于感染后约d15均自愈,LPS处理组小鼠的原虫血症水平在感染后d5-8显著低于对照组;（2）与对照组相比,在感染后d3和d5,LPS处理组小鼠脾脏DCs亚群表达水平显著升高,在感染后d3,TLR4表达水平显著升高。在感染后d5,LPS处理组小鼠脾脏细胞培养初始活化T表达水平显著升高。结论在P.y17×L感染早期,LPS可通过激活TLR4强化DC成熟进一步强化Th1免疫应答反应,降低原虫血症,这将为疫苗的开发提供新的靶点。     

鼠疟原虫感染大鼠和小鼠的种特异性分析

目的用约氏疟原虫(P.y) BY265株和伯氏疟原虫(P.b) ANKA-luciferase株感染小鼠和大鼠，分析鼠疟原虫感染宿主的种特异性。方法制备P.y和P.b的子孢子和裂殖子。分别用5×10⁴的P.y和P.b子孢子经尾静脉注射感染SD大鼠、BALB/c小鼠(P.y感染组，5只/组)和SD大鼠、C57BL/6J小鼠(P.b感染组，5只/组)，每日取尾静脉血涂片镜检，记录红内期疟原虫出现的时间。P.y子孢子感染SD大鼠和BALB/c小鼠后42 h取肝组织，实时荧光定量PCR (qRT-PCR)检测肝组织疟原虫18S rRNA的相对表达量。P.b子孢子感染SD大鼠和C57BL/6J小鼠后42 h活体成像法检测肝脏荧光值。分别用1×10⁸、1×10⁷、1×10⁶的P.y和P.b被感染红细胞(iRBC)感染SD大鼠(P.y感染组、P.b感染组)和易感小鼠(BALB/c、C57BL/6J)(阳性对照组)，每日取尾静脉血涂片镜检，计算原虫血症。分别用P.y、P.b裂殖子感染SD大鼠和易感小鼠，每日取尾静脉...     

基于16S rDNA高通量测序探讨肠道菌群对疟原虫感染小鼠的保护性作用

目的 探究Plasmodium berghei ANKA(P.bANKA)感染小鼠肠道菌群特异性改变及其保护性研究。方法 C57BL/6小鼠随机分为正常对照组(NC组)和P.bANKA感染组(P.b A组),动态留取粪便进行16S rDNA高通量测序。建立益生菌预处理组(Pre-probiotics+P.b A组),伊文思蓝灌注评估血脑屏障(BBB)完整性，苏木精-伊红(HE)染色，观察脑组织病理学改变。结果 16S rDNA高通量测序分析，P.bANKA感染小鼠后肠道菌群以乳酸杆菌和罗伊乳酸杆菌明显下降为主要变化趋势，且呈时间依赖性。动物验证实验发现，与P.b A组相比，Pre-probiotics+P.b A组原虫血症水平较低，小鼠于感染后第14 d开始出现死亡且未发生脑疟神经系统症状，BBB完整性较好，脑组织无明显损伤。结论 P.bANKA感染可明显改变宿主肠道菌群的结构和功能，其中以乳酸杆菌数量减少最为明显。外源性补充乳酸杆菌后可明显降低ECM小鼠原虫血症水平，提高生存率。其作用机制可能与保护BBB完整性，改善小鼠脑组织病理损伤密切相关。     

IL-12对伯氏疟原虫红内期感染小鼠淋巴细胞增殖活性的影响

目的探讨IL12对伯氏疟原虫(P.b)红内期感染小鼠淋巴细胞增殖活性的影响。方法每只BALB/c小鼠接种5×105个感染P.b的RBC,同时每天分别给予0.03或0.15μgIL12处理,用3HTdR掺入法检测脾淋巴细胞增殖转化活性,流式细胞仪测定T淋巴细胞亚群CD4+与CD8+百分比。结果每天给予0.03或0.15μgIL12处理,均可显著增加P.b感染小鼠脾淋巴细胞数量及CD4+、CD8+数。每天给予0.03μgIL12可显著促进脾淋巴细胞的增殖转化,而每天给予0.15μgIL12则明显抑制脾淋巴细胞增殖。结论适当低剂量IL12可促进P.b感染小鼠脾淋巴细胞增殖活性,增强机体抗感染免疫,而较大剂量IL12则抑制脾淋巴细胞增殖活性,甚至可致免疫病理损害。     

致死型约氏疟原虫感染BALB/c小鼠巨噬细胞的作用地位研究

摘 要：目的 固有免疫在疟疾急性期感染中的保护性作用地位有待深入研究。本研究旨在探讨巨噬细胞在致死型约氏疟原虫（Plasmodium yoelii 17XL,P.y 17XL) 感染中的作用地位。方法 雌性BALB/c在感染-2、0d腹腔注射氯磷酸脂质体（clodronate liposomes,300μl/只/次）,建立巨噬细胞消除模型。对照组小鼠在同时点注射同体积PBS脂质体。随后,小鼠腹腔注射1×106 P.y 17XL寄生的红细胞。计数原虫血症和观察生存率。ELISA法检测脾培养上清IFN-γ和IL-10水平,Griess法检测NO（NO2-）水平。脾细胞流式细胞术（FACS）染色,计数F4/80+巨噬细胞数量。结果 与对照组相比,巨噬细胞消除组原虫血症峰值提前出现,较对照组死亡时间提前。在感染0、3、5d,巨噬细胞消除组脾巨噬细胞占脾细胞百分含量和绝对值均明显降低。脾培养上清中IFN-γ、IL-10和 NO分泌水平明显下降。巨噬细胞消除后加速P.y 17XL感染BALB/c小鼠的感染进程且明显影响疟疾感染的最终结局。结论 巨噬细胞在P.y 17XL感染BALB/c小鼠中发挥重要的免疫保护作用。     

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.     

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.     

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.     

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.