外源性双链DNA诱导小鼠狼疮样肾损害的病理学特征

目的 观察马疫锥虫双链DNA(dsDNA)诱导的小鼠狼疮样肾脏损害的病理学特征.方法 将分离纯化的马疫锥虫动基体DNA(kDNA)与不完全弗氏佐剂乳化混合,以皮下途径对健康BALB/c小鼠进行免疫.8周后,检测相关生化与免疫学指标并观察其肾脏病理学表现;同时与BXSB小鼠和抗dsDNA抗体阳性狼疮肾炎(LN)患者进行比较.结果 经马疫锥虫kDNA免疫后小鼠的免疫学指标符合LN的特征,肾脏损害以肾病综合征为主要表现;与BXSB小鼠和LN患者相比较,其肾脏病理改变有一定相似性,但以Ⅱ型(系膜细胞增殖)与Ⅳ型(弥漫性增殖)等类型为主.结论 马疫锥虫dsDNA诱导的小鼠肾脏损害与抗dsDNA抗体阳性的LN损害相似,可以作为研究该类型LN的有效工具.     

IgG型抗双链DNA抗体对体外培养系膜细胞表型转化的影响

目的 探讨IgG型抗双链DNA(dsDNA)抗体对正常大鼠系膜细胞表型转化的影响.方法 将马疫锥虫动基体DNAI)~不同途径免疫大鼠,分别诱生IgG型、IgM型抗dsDNA抗体,其血清分别与正常大鼠系膜细胞(MCs)共同培养,同时设立正常血清对照组.采用酶联免疫吸附法(ELISA)与Western blot法检测Mcs表达转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、层黏连蛋白(LN)等水平,四甲基偶氮唑蓝(MTT)比色法检测MCs的增殖率,锥虫蓝染色法测活细胞比例,透射电镜下观察形态学改变.结果 与正常血清组比较,IgM抗体组的上述指标差异无统计学意义(P＞0.05),而:IgG抗体组的细胞增殖率与活细胞比例差异无统计学意义(P＞0.05),但其余指标皆上升(P＜0.05).电镜下IgG抗体组的Mcs呈肌成纤维细胞样形态改变,而其余两组大致正常.结论 IgG型抗dsDNA抗体可能通过诱导Mcs表型转化而参与狼疮肾炎的发病机制.     

大黄素抑制IgG型抗双链DNA抗体诱导的系膜细胞表型转化的研究

目的探讨大黄素对IgG型抗双链DNA（dsDNA）抗体诱导的大鼠系膜细胞（MCs）表型转化的抑制作用。方法利用马疫锥虫动基体DNA诱导大鼠产生IgG型抗dsDNA抗体，小牛胸腺DNA纤维素层析法提纯抗体。体外培养正常MCs（空白组），或在培养上清中分别加入IgG型抗dsDNA抗体、25mg／L大黄素（对照组），或同时加IgG型抗dsDNA抗体与大黄素（实验组），检测MCs合成转化生长因子-β1（TGF-β1）、α-平滑肌肌动蛋白（α-SMA）等水平，并在透视电镜下观察细胞显微结构的变化。结果与空白组比较，抗体对照组合成TGF-β1等水平明显升高（P〈0．05），并出现肌成纤维细胞样结构变化；而大黄素对照组的上述指标下降（P〈0．05），且细胞显微结构基本正常。与抗体对照组比较，实验组合成TGF-β1等水平降低，细胞显微结构大致正常。结论抑制IgG型抗dsDNA抗体诱导的系膜细胞表型转化可能是大黄素治疗狼疮肾炎的药理机制之一。     

含弓形虫ROP1基因真核表达重组质粒DNA免疫小鼠的研究Ⅲ.免疫鼠肌组织表达产物免疫酶法定位及血清IgG抗体测定

目的用所构建的弓形虫pcDNA3-ROP1真核表达重组质粒,经肌肉注射免疫小鼠,观察它在肌组织中的表达及不同免疫途径所诱导的体液免疫应答.方法碱裂解法大量制备pcDNA3-ROP1质粒,免疫BALB/c小鼠,每只鼠注射100ug, 两周后同量加强免疫一次,以pcDNA3空质粒及生理盐水组为对照.于免疫后第50天用间接免疫酶法检测注射局部肌组织重组蛋白的表达;ELISA法测定IgG抗体滴度.结果免疫鼠肌组织石蜡切片呈特异性阳性反应;血清IgG抗体90天后测定为阳性;皮下及肌肉不同免疫途径血清均显示阳性结果,无显著性差异.结论 pcDNA3-ROP1质粒DNA免疫小鼠后,肌组织内有重组蛋白表达,并能诱导机体产生IgG抗体.     

恶性疟原虫CSP基因重组蛋白及DNA免疫诱导小鼠体液免疫应答比较

本文利用恶性疟原虫FCC1/HN株环子孢子蛋白 (PfCSP)基因DNA质粒通过不同途径、不同剂量免疫小鼠 ,观察其产生的体液免疫应答反应 ,并将其与相应的重组表达蛋白疫苗进行比较。结果显示 :DNA免疫刺激机体产生抗体反应强度的免疫途径依次为肌肉、静脉和皮下 ;宿主对DNA免疫存在一定的剂量依赖性 ;ELISA和Dot -ELISA检测免疫后 4周和 7周 ,DNA质粒组刺激机体产生抗体的滴度均显著低于相应的重组蛋白组。表明PfCSPDNA疫苗与重组表达蛋白疫苗均可刺激小鼠产生特异性体液免疫应答 ,但前者诱导高滴度的抗体反应需要更长的时间     

弓形虫表面抗原P30 DNA疫苗免疫小鼠诱导体液免疫应答及保护性的初步观察

目的观察弓形虫表面抗原P30 DNA疫苗免疫BALB/c小鼠诱导其体内产生的体液免疫应答及抗虫感染的免疫保护作用. 方法重组质粒pBK-P30用生理盐水稀释后,肌注免疫BALB/c小鼠.分别于免疫后5周和10周,ELISA测定IgG抗体滴度;取免疫鼠的血液、肺、心脏、肝脏、脾、肾脏及肌肉PCR扩增P30基因;免疫鼠腹腔注射弓形虫速殖子攻击感染.结果两次检测,均可测到特异性IgG抗体,且抗体的滴度随免疫时间的延长而增高;免疫后5周,免疫鼠的上述组织均可扩增出P30基因条带,但免疫后10周仅血液扩增出特异P30基因条带;弓形虫速殖子腹腔攻击感染,免疫组鼠的平均存活时间较对照组鼠延长,但统计学差异不显著(P＞0.05). 结论弓形虫表面抗原P30 DNA疫苗能诱导BALB/c小鼠产生特异性体液免疫应答及部分抗虫免疫保护作用.     

多聚赖氨酸预处理花生DNA疫苗对花生蛋白诱导变态反应的作用

目的探讨经多聚赖氨酸预处理的花生DNA疫苗对花生蛋白诱导变态反应的预防作用。方法以编码花生蛋白Arah2的质粒DNA(pArah2)、经多聚赖氨酸(PLL)预处理的质粒DNA(PLL-pArah2)、磷酸盐缓冲液(PBS)和PLL对照皮内注射免疫Balb/c小鼠,1次/周,连续3周,再经腹腔注射纯化的Arah2蛋白免疫小鼠。在不同时间点收集各组小鼠血清,用酶联免疫吸附法检测血清中Arah2特异性IgG1、IgG2a抗体以及血清总IgE抗体水平,分析DNA疫苗免疫对蛋白诱导小鼠变态反应的影响。结果 pArah2及PLL-pArah2质粒DNA免疫后,小鼠血清Arah2特异性IgG1和IgG2a升高,但血清总IgE抗体未见升高。Arah2蛋白免疫后,不同处理组血清总IgE、Arah2特异性IgG1和IgG2a抗体水平均升高,但pArah2组血清总IgE和Arah2特异性IgG1抗体水平明显低于PBS对照;PLL-pArah2组血清总IgE和Arah2特异性IgG1、IgG2a抗体水平不仅明显低于PBS和PLL对照,且明显低于单纯pAra h2组。结论经多聚赖氨酸预处理的Arah2 DNA免疫能够更好地抑制之后蛋白免疫产生的变态反应应答,为DNA疫苗治疗过敏性疾病提供了新的思路。     

编码完整膜蛋白Sj23质粒DNA核酸疫苗免疫小鼠及其保护力的研究

目的 探讨用编码完整膜蛋白(Sj23)核酸疫苗诱导BALB/c小鼠抗日本血吸虫病保护力的作用.方法 大量制备DNA疫苗,BALB/c小鼠被分成3组,肌肉内注射进行免疫.免疫小鼠3次,间隔2周,末次免疫后2周,用ELISA法和Western Blotting法检测免疫鼠血清特异性抗体效价.结果 编码Sj2-pcDNA质粒免疫的小鼠产生了针对Sj23的特异性IgG,而peDNA对照组免疫小鼠血清则无此作用.Sj23-pcDNA疫苗对日本血吸虫攻击感染有一定的保护力.结论 Sj23-pcDNA疫苗能够诱导小鼠产生抗日本血吸虫攻击感染的保护力.     

恶性疟原虫裂殖子表面蛋白1 DNA与改良痘苗病毒组合疫苗诱导小鼠抗体应答

目的?摇探索DNA与改良痘苗病毒(MVA)组合免疫对增强恶性疟原虫裂殖子表面蛋白1（MSP1）抗体应答的作用。 方法 以人工合成MSP1全基因为基础分别构建DNA免疫质粒VR1020/190和重组MVA，单用VR1020/190或与表达质粒GM-CSF共同对小鼠进行初始免疫后，用重组病毒追加强化，采用DNA/MVA组合方案免疫BALB/c小鼠，ELISA测定血清IgG及其亚类水平，经腹腔接种转基因伯氏疟原虫Pb-PfM19进行攻击。 结果 DNA免疫能有效诱导小鼠产生抗MSP1-190抗体，其终点稀释度为1∶2 500，GM-CSF质粒共免疫组抗体的终点稀释度为1∶11 150，抗体亚类的测定表明GM-CSF质粒显著促进了IgG1类抗体应答， MVA追加可使单独免疫组和共免疫组抗体分别增加53和10倍；两实验组产生了水平相近的抗19 000抗体（1∶32 000），其含量占血清中MSP1总IgG的1/4-1/3。经转基因伯氏疟原虫Pb-PfM19攻击后小鼠的存活时间并没有明显延长（P＞0.05）。 结论 采用合成MSP1全基因进行DNA/MVA组合免疫可诱导小鼠产生显著的抗体应答，抗体的详细特性和保护作用正在进一步研究中。     

静脉应用核小体H2B表位肽防治小鼠实验性系统性红斑狼疮的研究

目的 探讨静脉途径应用基于核小体的表位肽防治系统性红斑狼疮(SLE)的可行性。方法 以凋亡细胞诱导的SLE样症状小鼠为模型，用核小体表位肽H2B14-28静脉途径诱导小鼠免疫耐受，观察实验小鼠自身抗体产生情况以及SLE样临床症状改善情况。结果用表位肽H2B14-28诱导免疫耐受以后，可显著降低实验小鼠抗核抗体(ANA)、抗双链DNA(dsDNA)抗体、抗RNP抗体和抗心磷脂(ACL)抗体的产生，同时显著改善小鼠蛋白尿、白细胞低下、肾脏免疫复合物沉积程度和病理损伤程度。结论 静脉途径应用核小体表位肽H2B14-28可有效阻止实验性SLE病变的形成，基于核小体表位肽的肽疫苗的研究有可能为SLE的防治开辟一条全新的思路和途径。     

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.     

Language of CTO interventions – Focus on hardware

The knowledge of variety of chronic total occlusion (CTO) hardware and the ability to use them represents the key to success of any CTO interventions. However, the multiplicity of CTO hardware and their physical character and the terminology used by experts create confusion in the mind of an average interventional cardiologist, particularly a beginner in this field. This knowledge is available but is scattered. We aim to classify and compare the currently used devices based on their properties focusing on how physical character of each device can be utilized in a specific situation, thus clarifying and simplifying the technical discourse.