胰岛素预防NOD鼠糖尿病的机制研究

目的观察胰岛素（Ins）对非肥胖糖尿病（NOD）小鼠胰岛炎和糖尿病的影响，并探讨其诱导免疫耐受的机制。方法60只NOD雌鼠随机分为胰岛素（NPH）处理组和PBS对照组，分别于第4、12、20、28周予皮下注射NPH6U（60μl）及PBS60μl十不完全弗氏佐剂60μl。于12周龄观察胰岛炎和胰岛β细胞凋亡；测定血清和脾细胞上清IL-4和干扰素（IFN）γ浓度；检测胰岛内和脾细胞IL-4、IFN-γ mRNA的表达；并行联合过继转移实验。结果Ins组糖尿病发病率和β细胞凋亡率比PBS组低。血清和脾细胞上清IL-4浓度及胰岛内和脾细胞IL-4mRNA表达较PBS组高，而IFN-γ浓度及IFN—γ mRNA表达比PBS组低。过继转移实验Ins组DM发病率比PBS组低。结论Ins能诱导NOD鼠调节T细胞的产生，使全身和胰岛局部T细胞由Th1向Th2转型，抑制β细胞凋亡，从而预防DM．     

吡格列酮对NOD鼠糖尿病的预防作用及其机制探讨

目的探讨吡格列酮（PIO）对NOD鼠糖尿病发病率和胰岛炎的影响及其作用机制。方法4周龄NOD雌鼠随机分为2组，分别摄食0．02％PIO混合饲料（PIO，n=26）和普通饲料（对照组，n=25），观察30周龄时的糖尿病累积发病率。各组另取12周龄未患病NOD鼠（n=15）胰腺H～E染色观察胰岛炎；ELISA法测血清、脾细胞培养上清干扰素γ（IFN-γ）和白细胞介素4（IL～4）水平；RT-PCR检测脾脏IL-4、IFN～γmRNA的表达水平。结果30周龄时，PIO组发病率较对照组明显降低（P〈0．05）。12周龄时，PIO组胰岛炎平均积分低于对照组（P〈0．05）；血清、脾上清IL～4水平，脾脏IL-4mRNA表达水平显著性高于对照组（P〈0．05）；PIO血清、脾上清IL-4／IFN-γ比值水平高于对照组（P〈0．05）。结论PIO通过上调IL～4水平，促使免疫平衡向Th2方向偏移，从而使NOD鼠胰岛炎减轻，而在一定程度上预防和延缓NOD鼠糖尿病的发生。     

完全费氏佐剂预防非肥胖糖尿病鼠胰岛炎和糖尿病的效应

目的 探讨完全费氏佐剂（CFA）在预防非肥胖糖尿病（NOD）鼠胰岛炎和糖尿病机制中的作用。方法 将3周龄NOD雌鼠随机分入CFA组、磷酸盐缓冲液（PBS）对照组,分别给予后足板一次性注射CFA50μl、PBS50μl,于6周龄、12周龄、30周龄时观察胰岛β细胞凋亡、胰岛炎程度和糖尿病发病率。结果 6周、12周时CFA组胰岛炎积分、β细胞凋亡率均明显低于周期PBS组（P＜0.05,P＜0.01）,6周龄、12周龄胰岛炎积分与β细胞凋亡率存在正相关。到30周龄时,CFA组11只鼠只有1只发生糖尿病（1／11）,明显低于PBS组[11只中有9只（9／11）] ,P=0.001。结论 早期给予NOD鼠CFA可明显减轻其胰岛炎和糖尿病的发生,其机制与抑制β细胞凋亡有关。     

NOD小鼠胰岛β细胞凋亡及完全弗氏佐剂的预防效应

目的　探讨NOD小鼠胰岛 β细胞凋亡的规律及完全弗氏佐剂 (CFA)在预防胰岛炎和糖尿病中的作用。方法　计数 12天龄、6周龄、12周龄未处理的NOD雌鼠胰岛 β细胞的凋亡。将 3周龄NOD雌鼠随机分入CFA组、PBS对照组 ,分别给予后足板一次性注射CFA 5 0 μl、PBS 5 0 μl,于 6周龄、12周龄、30周龄时 ,观察胰岛 β细胞凋亡率 ,胰岛炎程度和糖尿病发病率。 结果　未处理组 12天龄胰岛 β细胞凋亡率分别高于 6周龄、12周龄 (P <0 .0 1)。 6周、12周时CFA组胰岛炎积分 ,β细胞凋亡率均明显低于同期PBS组 (P <0 .0 5 ,P <0 .0 1) ,6周龄、12周龄胰岛炎积分与 β细胞凋亡率存在正相关 (r =0 .5 9,P <0 .0 1)。到 30周龄时 ,CFA组糖尿病发病率 (9.1% )较PBS组 (81.8% )明显降低 (P=0 .0 0 1)。结论　早期给予NOD鼠CFA可明显减轻胰岛炎和预防糖尿病的发生 ,其机制与抑制 β细胞凋亡有关。     

完全弗氏佐剂预防NOD鼠糖尿病的机制及其与细胞因子Th表达和β细胞凋亡的关系

目的　探讨完全弗氏佐剂 (CFA)预防非肥胖糖尿病 (NOD)小鼠胰岛炎与糖尿病的作用机制。　方法　 42只NOD雌鼠随机分为CFA处理组 (n =2 1)和磷酸盐缓冲液 (PBS)对照组 (n =2 1) ,3周龄时分别于后脚板一次性注射 50 μlCFA和PBS。于 12周龄时观察胰岛炎的程度、胰岛Fas和FasL的表达、β细胞凋亡情况以及测定血清白细胞介素 4(IL 4)、γ干扰素 (Ifn γ)水平和胰岛内IL 4、Ifn γ、白细胞介素 1β(IL 1β)、FasmRNA的表达水平。 　结果　 12周龄时CFA组胰岛炎积分和β细胞凋亡率均显著低于PBS组 (P均 <0 .0 5)。Fas只在PBS组胰岛内表达 ,而FasL在CFA和PBS组均有相似表达。 12周龄CFA组血清IL 4水平较PBS组增高而Ifn γ水平降低 (P均 <0 .0 5) ;RT PCR结果显示 ,CFA组胰岛内IL 4mRNA转录水平较对照组显著增高 (P <0 0 1) ,而Ifn γ、IL 1β、FasmRNA转录水平则明显降低 [P均 <0 .0 1(Ifn γ ,IL 1β) ,P <0 .0 5(Fas) ]。 3 0周龄时CFA组糖尿病发病率 (9.1% ,1/ 11)比PBS组 (81.8% ,9/ 11)显著降低 (P =0 .0 0 1)。　结论　CFA预防NOD鼠胰岛炎及糖尿病可能与胰岛自身反应T细胞由Th1向Th2转型 ,从而抑制Fas介导的 β细胞凋亡相关     

完全弗氏佐剂预防非肥胖糖尿病鼠胰岛炎和糖尿病的效应

目的探讨完全弗氏佐剂(CFA)在预防非肥胖糖尿病(NOD)鼠胰岛炎和糖尿病机制中的作用.方法将3周龄NOD雌鼠随机分入CFA组、磷酸盐缓冲液(PBS)对照组,分别给予后足板一次性注射CFA 50 μl、PBS 50 μl,于6周龄、12周龄、30周龄时观察胰岛β细胞凋亡、胰岛炎程度和糖尿病发病率.结果 6周、12周时CFA组胰岛炎积分、β细胞凋亡率均明显低于同期PBS组(P＜0.05, P＜0.01), 6周龄、12周龄胰岛炎积分与β细胞凋亡率存在正相关.到30周龄时,CFA组11只鼠只有1只发生糖尿病(1/11),明显低于PBS组[11只中有9只(9/11)],P=0.001.结论早期给予NOD鼠CFA可明显减轻其胰岛炎和糖尿病的发生,其机制与抑制β细胞凋亡有关.     

IL-1β、IFN-γ对胰岛β细胞凋亡及胰岛素释放的影响

体外分离培养SD大鼠胰岛细胞，分别或共同加入IL-1β（终浓度20u／mL）和IFN-γ（终浓度150u／mL）处理。加入11．1mM的葡萄糖刺激胰岛素释放。应用放射免疫分析法测定培养液上清中胰岛紊浓度，流式细胞术检测胰岛细胞凋亡。结果：IL-1β单独作用于胰岛细胞，能抑制高糖刺激下胰岛素分泌及促进胰岛细胞凋亡。但与对照组比较差异无显著性（P〉0．05）；IFN-γ单独或联合IL-1β能显著抑制胰岛紊分泌（P〈0．05），促进胰岛细胞凋亡（P〈0．05）。结论：IFN-γ和IL-β能诱导胰岛β细胞凋亡，并且有协同作用。与自身免疫性糖尿病的发生、发展有一定关系。     

吡格列酮预防非肥胖糖尿病鼠胰岛β细胞凋亡的机制研究

目的 探讨吡格列酮预防非肥胖糖尿病小鼠胰岛β细胞凋亡的机制.方法 (1)将4周龄NOD雌鼠分为吡格列酮(21只)及对照(21只)组,分别摄食含0.02％吡格列酮的混合饲料和普通营养饲料.观察52周龄的累积糖尿病发病率.(2)各组取12周龄未发病NOD鼠(n=15)胰腺,HE染色观察胰岛炎;TUNEL+SABC法检测胰岛β细胞凋亡.(3)ELISA法测定血清、脾细胞培养上清IFN-γ和IL-4水平及培养脾细胞核因子PPARγ、NF-κB活性.结果 (1)30、52周龄时,吡格列酮及对照组发病率分别为57.1％和76.2％ 、76.2％和90.5％(均P＞0.05);15周龄时,吡格列酮及对照组发病率分别为4.8％和33.3％(P=0.045).(2) 12周龄时,吡格列酮组正常胰岛和胰岛周围炎比例(14.73％,26.02％)高于对照组(5.69％,15.72％;均P＜0.01),胰岛内炎比例(59.25％)则低于对照组(78.59％,P＜0.01);吡格列酮组胰岛β细胞凋亡率(6.17％±3.62％)低于对照组( 10.62％±4.43％,P=0.008).(3)12周龄NOD鼠吡格列酮组血清IFN-γ水平[(561.05 ±78.61) pg/ml]显著低于对照组[(666.43±28.42) pg/ml,P=0.045];在培养的脾细胞上清中,吡格列酮组IFN-γ水平[(605.84±65.60) pg/ml]显著低于对照组[(692.20±44.98)pg/ml,P=0.041].(4)在培养的脾细胞中,吡格列酮组PPARγ活性(0.06±0.01)高于对照组(0.03±0.01,P=0.013),NF-κB活性(0.03±0.01)较对照显著降低(0.08±0.01,P=0.001).结论 吡格列酮活化PPARγ,抑制NF-κB活性,血清和脾细胞上清IFN-γ下降,Th细胞向Th1方向分化减少,NOD鼠胰岛炎减轻、胰岛β细胞凋亡减少.     

NOD鼠口服胰岛素诱导免疫耐受的疗效及形态学观察

目的 评价口服胰岛素诱导免疫耐受在防治自身免疫性糖尿病方面的效果。方法 雌性NOD鼠120只，随机分为胰岛素组和对照组各60只，6周龄始分别口服猪胰岛素500ul或磷酸盐缓冲液（PBS）500ul，至40周龄。6、12、20、25、30周龄分别测定两组未发病NOD鼠血清胰岛素（insulin)水平，并观察胰岛形态学变化。结果 40周龄糖尿病累计发生率对照组为91％，胰岛素组为11％，两者具有显著性差异（P＜0．01）。未发病NOD鼠中，20、25、30周龄对照组胰岛炎明显重于胰岛素组，血清胰岛素水平显著低于胰岛素组（P均＜0．05）。发病的NOD鼠中，对照组NOD糖尿病鼠血清胰岛素水平显著低于胰岛素组糖尿病鼠（P＜0．05）。结论 口服胰岛素可明显延迟NOD鼠糖尿病的发病时间，降低糖尿病发生率，缓解动物发病后的病情。     

己酮可可碱预防NOD鼠1型糖尿病的机理研究

目的 探讨己酮可可碱（Pentoxifylline,PTX)对非肥胖糖尿病（NOD）小鼠1型糖尿病发病率，胰岛素的影响及其机制。方法 采用动物模型NOD鼠，注射环磷酰胺（CP）加速其发病。给PTX药物后计算糖尿病发病率，HE染色观察胰岛炎，并用逆转录（RT）PCR法检测脾细胞干扰素γ（IFN－γ），肿瘤坏死因子α（TNF－α），白介素10（IL－10）mRNA的表达。结果 PTX组糖尿病发生率（30．00％）明显低于对照组（67．86％）（P＜0．1）；胰岛炎程度也明显减轻（P＜0．001）；脾细胞IFN－γ，TNF－αmRNA的表达较对照组明显降低（P＜0．05），IL－10mRNA的表达无明显改变。结论 PTX可预防NOD鼠发生糖尿病，其机制可能与纠正Th1与Th2型细胞因子比例失衡有关。     

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.     

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.