乙型肝炎患者外周血CD4~+T细胞Runx3的表达及意义

目的探讨乙型肝炎患者外周血CD4+T细胞Runx3 mRNA的表达及意义。方法应用实时定量PCR检测37例慢性乙型肝炎(chronic hepatitis B,CHB)患者(CHB组),11例急性乙型肝炎(acute hepatitis B,AHB)患者(AHB组)和19名健康对照者(健康对照组)外周血CD4+T细胞Runx3 mRNA的表达水平。结果 CHB组Runx3 mRNA的表达水平明显低于AHB组和健康对照组(P均0.01)。AHB组Runx3 mRNA的表达水平明显高于健康对照组(P0.05)。CHB重度组Runx3 mRNA的表达水平明显高于CHB轻度组(P0.05)。结论 Runx3在CHB的发病过程中可能发挥了重要的作用,CHB患者Runx3表达水平降低可能与其Th1/Th2失衡有关。     

Notch信号通路对慢性丙型肝炎患者CD8~+T淋巴细胞功能的影响

目的观察Notch信号通路对慢性丙型肝炎患者CD8~+T淋巴细胞功能的影响,初步阐释Notch信号通路在慢性丙型肝炎发病中的作用。方法收集2016年1月-8月在陕西省人民医院就诊的慢性丙型肝炎初治患者38例和健康志愿者17例,分选CD8~+T淋巴细胞,应用实时定量PCR法检测CD8~+T淋巴细胞中Notch1和Notch2 mRNA的表达水平。分别应用直接接触培养系统和间接接触培养系统将从HLA-A2限制性丙型肝炎患者分选的CD8~+T淋巴细胞与HCV感染的Huh7.5细胞共培养,加入Notch信号通路抑制剂DAPT,通过检测乳酸脱氢酶和细胞因子水平分析CD8~+T淋巴细胞的细胞毒性和非细胞毒性功能的变化。2组间计量资料的比较采用t检验。结果 Notch1和Notch2 mRNA在慢性丙型肝炎患者CD8~+T淋巴细胞中的表达水平较健康志愿者均显著升高,分别升高约10倍和4倍,差异均有统计学意义(t值分别为6.63、18.88,P值分别为0.003、0.001)。抑制Notch信号通路可增加直接接触培养系统中细胞死亡的比例[(26.41±4.76)%vs(13.65±4.65)%,t=3.32,P=0.029];但抑制Notch信号通路对间接接触培养细胞中CD8~+T淋巴细胞毒性无明显影响。抑制Notch信号通路后,IFNγ的表达在直接接触培养系统[(52.19±12.38)pg/ml vs(18.62±5.66)pg/ml,t=4.27,P=0.013]和间接接触培养系统[(18.86±3.09)pg/ml vs(5.52±2.52)pg/ml,t=5.79,P=0.004]中均显著升高。TNFα的表达在抑制Notch信号通路后仅在直接接触培养系统中显著升高[(1296.0±293.3)pg/ml vs(585.3±32.62)pg/ml,t=4.17,P=0.014],而在间接接触培养系统中无明显变化[(297.3±175.4)pg/ml vs(273.7±105.9)pg/ml,t=0.20,P=0.851]。结论抑制Notch信号通路可增强慢性丙型肝炎患者中CD8~+T淋巴细胞的细胞杀伤和非细胞杀伤功能,Notch信号通路在慢性HCV感染中可能通过抑制CD8~+T淋巴细胞功能发挥诱导免疫耐受的作用。     

乙型肝炎患者外周血CD4+ CD25+调节性T细胞表型与功能分析

目的 观察急、慢性乙型肝炎(AHB、CHB)患者外周血CD4+CD25 high调节性T细胞(Treg)的频率、表型和功能特点.方法 采集16例AHB急性发病期(发病后第1周)患者、72例CHB患者和32例健康人的外周血,检测Treg频率,并分析其表面CD45RO、CD45RA、HLA-DR、CD95和细胞内细胞毒T淋巴细胞相关抗原4(CTLA-4)的表达水平.应用实时荧光定量RT-PCR检测CD4+ CD25+、CD4+ CD25-、CD4+和CD4-等细胞亚群和外周血单个核细胞(PBMC)的FoxP3 mRNA表达量.通过MACS免疫磁珠分选Treg,并应用[3H]掺入法检测Treg抑制抗-CD3抗体和HBV抗原刺激的PBMC增殖能力,并观察Treg对HBV抗原或抗-CD3抗体刺激自体PBMC分泌IFNγ的影响.结果 CD4+CD25 high Treg高表达CD45RO、HLA-DR、CD95和细胞内CTLA-4,低表达CD45RA,并且较特异的高表达FoxP3 mRNA.乙型肝炎病人外周血Treg频率与健康对照(3.50±0.72)%比较无统计学差异,但CHB组(3.90±1.44)%显著高于AHB组(3.10±0.87)%,P＜0.05.Treg本身对于HBV抗原或抗-CD3抗体刺激没有明显的增殖反应和IFNγ分泌,但可抑制自体PBMC增殖和IFNγ分泌,其中对HBV抗原刺激引起的细胞反应抑制作用较强.结论 HBV感染者外周血Treg较特异地表达FoxP3分子,能抑制HBV抗原特异性细胞免疫反应,这对于深入阐明CHB发病机制具有重要意义.     

慢性乙型肝炎患者血清转化生长因子β1与CD4~+CD25~+Foxp3~+调节性T淋巴细胞的关系

目的 调查慢性乙型肝炎患者转化生长因子β1(TGF β1)与CD4~+CD25~+Foxp3~+调节性T淋巴细胞异常的关系.方法 将实验对象分为慢性乙型肝炎(CHB)患者、无症状HBV携带者(AsC)、乙型肝炎恢复者、正常人对照4组.通过流式细胞术分析外周CD4~+CD25~+Foxp3~+调节性T淋巴细胞的表型和频率以及实时荧光定量PCR分析Foxp3表达水平;并通过酶联免疫吸附法检测血清中TGF β1水平.根据数据不同采用方差分析或秩和检验进行统计学分析.结果 CHB或AsC组外周CD4~+T淋巴细胞中CD4~+CD25~+Foxp3~+T淋巴细胞频率以及CD4~+CD25~+T淋巴细胞中Foxp3 mRNA表达水平显著高于正常人对照组,差异有统计学意义(P<0.05).CHB患者和AsC血清中TGF β1水平显著高于正常人和乙型肝炎恢复者,并与CD4~+CD25~+Foxp3~+T淋巴细胞频率存在显著正相关(r=0.78,P<0.01).结论 CHB和AsC患者TGF β1与CD4~+CD25~+Foxp3~+调节性T淋巴细胞增高有密切关系.     

乙型肝炎病毒感染不同阶段患者外周血CD4+CD25+调节性T细胞的测定

目的 测定HBV感染不同阶段患者外周血CD4+CD25+调节性T细胞(Treg)的频率及标记分子,并分析其与临床指标的相关性.方法 采集79例慢性乙型肝炎(CHB)、12例急性乙型肝炎(AHB)患者、26例无症状HBV携带者(ASC)和20例健康对照的外周血,流式细胞仪分析Treg频率、Treg细胞表面和胞内特征性分子的表达.普通RT-PCR和相对荧光定量PCR测定叉头/翼状转录因子3(Foxp3)在CD25+Treg细胞的表达水平.所有患者及健康对照均经ELISA检测HBV血清标记物水平,实时荧光定量PCR测定血清HBV DNA载量,并进行肝功能检测.结果 总CD4+CD25+T细胞占外周血CD4+T细胞的比率,在各患者组和健康对照组之间差异均无统计学意义(P＞0.05).HBeAg阳性CHB组CD4+CD25高表达T细胞频率(3.42%±0.81%)与HBeAg阴性CHB组(3.19%±0.67%)、ASC组(3.05%±0.64%)比较,差异无统计学意义(均P＞0.05),但明显高于健康对照组(2.72%±0.71%,P=0.034)和AHB组(2.25%±0.54%,P=0.013).CD4+CD25高表达T细胞表面高表达CD45RO、CD25分子,低表达CD45RA,细胞内高表达细胞毒性T淋巴细胞相关抗原-4(CTLA-4)和Foxp3.各患者组及健康对照组Treg中Foxp3表达水平的差异无统计学意义(P＞0.05).CHB组患者的Treg频率与血清病毒载量呈正相关(r=0.48,P=0.018).结论 Treg可能通过抑制T细胞免疫应答反应而影响病毒清除,并与CHB患者的持续感染密切相关.     

慢性HBV感染者外周血T淋巴细胞亚群的变化及其与白细胞介素15、白细胞介素16的相关性分析

目的分析慢性HBV感染者外周血T淋巴细胞(CD3+、CD4+、CD8+和CD4+/CD8+)及细胞因子白细胞介素(IL)15、IL-16表达水平的变化及临床意义。方法选择2014年7-12月在兰州市第二人民医院治疗的慢性乙型肝炎患者63例(CHB组),慢性HBV携带者112例(ASC组),健康对照者84例(对照组),运用流式细胞术检测3组患者血清中T淋巴细胞亚群水平,酶联免疫法检测IL-15、IL-16表达水平,运用聚合酶链反应检测HBV DNA载量。多组间比较采用方差分析,组内数据的相关分析采用Pearson相关法。结果与健康对照组相比,ASC组和CHB组CD3+、CD4+T淋巴细胞百分数以及CD4+/CD8+T淋巴细胞百分数比值均显著降低,CD8+T淋巴细胞百分数显著升高,差异均有统计学意义(P值均0.05);ASC组和CHB组IL-15、IL-16表达水平均显著高于健康对照组,差异均有统计学意义(P值均0.05)。CHB患者中HBV DNA高载量组CD3+、CD4+、CD4+/CD8+T淋巴细胞百分数比值较HBV DNA低载量组显著降低,CD8+T淋巴细胞百分数显著升高,差异均有统计学意义(P值均0.05);ASC组HBV DNA高载量者CD4+、CD4+/CD8+T淋巴细胞百分数比值显著低于HBV DNA低载量组,CD8+T淋巴细胞百分数显著高于HBV DNA低载量组,差异均有统计学意义(P值均0.05)。CHB患者外周血CD4+表达水平与IL-15呈正相关(r=0.516,P0.05),CD8+表达水平与IL-16呈正相关(r=0.665,P0.05)。ASC组外周血CD3+表达水平与IL-15呈正相关(r=0.618,P0.05)。结论 HBV感染者存在T细胞亚群比例异常,IL-15、IL-16可能导致机体存在不同程度细胞免疫功能障碍和细胞免疫调节异常。     

microRNA-18a通过PPARα/γ信号通路上调慢性乙型肝炎患者调节性免疫功能

目的分析microRNA-18a(miRNA-18a)通过过氧化物酶体增殖物激活受体α/γ(PPARα/γ)信号通路对慢性乙型肝炎患者调节性免疫功能的影响。方法纳入空军特色医学中心(原中国人民解放军空军总医院)2017年4月-2018年10月98例慢性乙型肝炎患者和96例非乙型肝炎患者分别作为试验组和对照组。实时荧光定量聚合酶链反应检测2组患者血清中的miRNA-18a mRNA的相对表达量;流式细胞技术检测人外周血单个核细胞中miRNA-18a的表达;酶联免疫吸附试验检测miRNA-18a对CD4~+CD25~+调节性T淋巴细胞(Treg)相关细胞因子分泌水平的影响;Western Blot检测2组患者PPARα/γ信号通路相关蛋白表达。再将PBMC分为si-miRNA-18a抑制组(转染miRNA-18a抑制剂)和si-miRNA-18a正常对照组(转染miRNA-18a质粒),用流式细胞技术检测抑制miRNA-18a表达对CD4~+CD25~+Treg细胞频率的影响,Western Blot检测2组PPARα/γ信号通路相关蛋白的表达。正态分布的计量资料2组间比较采用t检验。Pearson相关分析检验miRNA-18a表达与PPARα/γ信号通路相关蛋白的相关性。结果试验组miRNA-18a mRNA表达水平在血清和肝组织中较对照组均显著上调(t值分别为9.634、9.863,P值均0. 01)。试验组CD4~+CD25~+Treg细胞频率较对照组显著上调(t=9.854,P0. 01)。试验组的IFNγ分泌水平和IL-9分泌水平较对照组均明显上调(t值分别为8.235,8.382,P值均005)。试验组的PPARα和PPARγ表达水平较对照组均明显上调(t值分别为4.229、3.545,P值均0. 05)。si-miRNA-18a抑制组外周血CD4~+CD25~+Treg细胞占CD4~+T淋巴细胞的比例明显低于si-miRNA-18a正常对照组(t=3.968,P0. 01)。si-miRNA-18a抑制组的PPARα和PPARγ表达水平均明显较si-miRNA-18a正常对照组低(t值分别为5.023、4.983, P值均0. 05)。miRNA-18a与PPARα/γ信号通路中的PPARα及PPARγ蛋白表达水平均呈正相关(r值分别为0.701、0.682,P值均0. 05)。结论 miRNA-18a可能通过激活PPARα/γ信号通路对慢性乙型肝炎患者调节性免疫功能产生影响,使其免疫功能相关细胞表面因子频率和细胞因子分泌水平上调。     

慢性乙型肝炎患者外周血中T淋巴细胞亚群和CD4~+CD25~+调节性T淋巴细胞的表达水平与HBV DNA定量的相关分析

目的探讨慢性乙型肝炎(CHB)患者外周血中T淋巴细胞亚群(CD3+T、CD4+T、CD8+T、CD4+/CD8+T)和CD4+CD25+调节性T淋巴细胞(Treg)表达水平与HBV DNA定量的相关性。方法选取2012年7月-12月在兰州市第二人民医院就诊的175例HBV感染者作为研究对象,分为CHB组(63例)、HBV携带者(ASC组)(112例),以同期50例健康体检者作为对照组。采用流式细胞术检测3组患者血清中的CD3+T、CD4+T、CD8+T、CD4+T/CD8+T和CD4+CD25+Treg表达水平,实时荧光定量PCR法检测HBV DNA载量。多组间均数比较采用方差分析,进一步两两比较采用LSD-t检验,计数资料组间比较采用χ2检验;采用Pearson进行相关性分析。结果 CHB患者外周血CD4+CD25+Treg表达水平(30.97±18.78)%显著高于对照组(27.30±17.59)%,差异具有统计学意义(P0.05);与对照组相比,CHB组、ASC组的CD3+T、CD4+T淋巴细胞百分数和CD4+与CD8+T淋巴细胞百分数比值显著降低,差异均有统计学意义(P值均0.05),CD8+T淋巴细胞百分数显著升高,差异均有统计学意义(P值均0.05);ASC组和CHB组CD4+CD25+Treg表达水平与HBV DNA呈正相关(r值分别为0.501、0.682,P值均0.01)。结论 HBV感染者存在T淋巴细胞亚群比例异常,HBV DNA可以促进CD4+CD25+Treg表达水平升高,说明CD4+CD25+Treg和CD4+与CD8+T淋巴细胞比例失衡在乙型肝炎慢性化过程中发挥重要作用。     

不同病毒载量HBV感染者外周血T淋巴细胞亚群和转化生长因子β的变化及意义

目的通过检测不同病毒载量慢性乙型肝炎(CHB)患者和HBV携带者(ASC)外周血T淋巴细胞各亚群变化的规律,探讨T淋巴细胞与人转化生长因子(TGF)β、ALT及TBil相关性。方法选择2012年7月至2012年12月兰州市第二人民医院收治的175例HBV感染者作为研究对象,分为ASC组(112例)、CHB组(63例),以同期健康体检者84例作为对照组。运用流式细胞术检测3组血清中T淋巴细胞亚群水平,同时检测肝功能指标等。正态分布的计量资料多组间比较采用方差分析,偏态分布的计量资料组间比较采Kruskal-Wallis H秩和检验。结果与健康对照组相比,ASC和CHB组TGFβ表达水平显著升高(P均0.05),CD3+、CD4+T淋巴细胞百分数显著降低(P0.05或P0.01),CD4+与CD8+T淋巴细胞比值显著降低(P0.01),CD8+T淋巴细胞百分数显著升高(P0.05或P0.01)。ASC与CHB组HBeAg阳性组和HBeAg阴性组患者CD3+、CD4+T淋巴细胞百分数均降低,CD8+T淋巴细胞百分数升高,CD4+与CD8+T淋巴细胞百分数降低。结论 TGFβ可能参与了CHB的发病过程,其免疫抑制作用可能是通过对T淋巴细胞、抗原递呈细胞成熟分化的抑制等多个环节发挥作用。     

调节性T淋巴细胞对成人乙型肝炎疫苗接种免疫应答的影响

目的 通过对乙型肝炎疫苗接种后不同免疫应答人群调节性T淋巴细胞(Treg细胞)Foxp3 mRNA的表达和细胞因子分泌的检测,探讨乙型肝炎疫苗接种后免疫应答与免疫调节细胞和细胞因子之间的内在联系.方法 采集不同反应人群全血,实时荧光定量PCR检测人外周血单个核细胞Foxp3 mRNA的表达;流式细胞术对外周血单核细胞的表面标志物CD4、CD25进行检测;酶联免疫法检测外周血单个核细胞植物血凝素(PHA)和HBsAg刺激后白细胞介素(IL)-4、IL-12、IL-18、干扰素(IFN)γ的产生水平.根据不同资料采用方差分析、q检验或相关分析进行统计学处理.结果 (1)PHA和HBsAg刺激前后,无应答组Foxp3的表达均高于应答组和对照组,差异有统计学意义(P＜0.05).(2)应答组外周血CD4+CD25+Treg细胞占CD4+T淋巴细胞的百分比(0.59%±0.46%)明显低于对照组(1.30%±1.44%),差异有统计学意义(F=2.990,P＜0.01).(3)各组外周血单个核细胞经PHA和HBsAg刺激后,无应答组的IFN γ浓度[(11.00±9.03)U/ml]明显低于应答组[(38.88±28.16)U/ml],差异有统计学意义(P＜0.01).(4)各组外周血单个核细胞经PHA和HBsAg刺激后,IL-18、IL-4、IL-12的浓度差异均无统计学意义.结论 CD4+CD25+Foxp3+Treg细胞在一定程度上参与了乙型肝炎疫苗接种应答的负性调控;乙型肝炎疫苗接种后无应答与IFN γ分泌不足有关;抗-HBs滴度水平与IFN γ和CD4+CD25+Foxp3表达无相关性.     

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.