日本血吸虫SjC23 DNA疫苗基因枪免疫小鼠诱导免疫保护作用的研究

目的研究日本血吸虫中国大陆株23 kDa膜蛋白DNA疫苗基因枪免疫诱导BALB/c小鼠产生的抗血吸虫感染作用.方法60只雌性BALB/c小鼠随机分为6组:pcDNA3.1组(对照组),每只小鼠用基因枪经腹部皮肤免疫pcDNA3.1质粒DNA 2次,共2 μg;SjC23基因枪(gg)组,每鼠用基因枪免疫pcDNA3.1-SjC23质粒DNA 2 μg;SjC23肌注(im)组,每鼠肌注100 μg pcD-NA3.1-SjC23质粒DNA;SjC23/CpG(gg)组,每鼠用基因枪免疫pcDNA3.1-SjC23/CpG质粒DNA2μg;SjC23/CpG(im)组,每鼠肌注100μg pcDNA3.1-SjC23/CpG质粒DNA;联合免疫组,每只小鼠先肌注100μg pcDNA3.1-SjC23/CpG质粒DNA,第2天用基因枪免疫2 μg.各组小鼠隔2周加强免疫1次,共3次.末次免疫后4周每鼠经腹部皮肤攻击感染(45±1)条日本血吸虫尾蚴,45 d后剖杀,计数成虫及肝脏虫卵数.首次免疫前2天及感染前2天经尾静脉采血检测IgG抗体水平及抗体亚类IgG1、IgG2a.末次免疫后3周检测脾细胞经ConA和rSjC23-HD刺激后培养上清中鼠IL-2、IL-4和IFN-γ的水平.结果SjC23(gg)组、SjC23/CpG(gg)组及联合免疫组小鼠所检成虫数均低于对照组(P均＜0.01),减虫率分别为19.57%、25.38%和32.31%;SjC23(gg)组、SjC23/CpG(gg)组及联合免疫组小鼠检获虫卵数均低于对照组(P均＜0.05),减卵率分别为16.92%、19.56%和27.73%.SjC23(im)组和SjC23/CpG(im)组分别获得了28.07%和35.14%的减虫率及21.56%和26.52%的减卵率.抗体检测结果,SjC23(gg)组、SjC23/CpG(gg)组、SjC23(im)组、SjC23/CpG(im)组和联合免疫组均诱导小鼠产生了特异性IgG抗体.SjC23(gg)组IgG1的A450值为0.102,而IgG2a几乎检测不到;SjC23/CpG(gg)组IgG2a/IgG1比值为2.01;联合免疫组IgG2a/IgG1比值为5.18;SjC23(im)组和SjC23/CpG(im)组IgG2a/IgG1的比值分别为10.06和12.15.脾细胞经ConA和rSjC23-HD刺激,联合免疫组检测到较高水平的IL-2.SjC23(gg)组小鼠脾细胞经特异及非特异性抗原刺激均产生较高水平的IL-4.脾细胞经ConA刺激,IFN-7的水平SjC23/CpG(gg)组较SjC23(gg)组有所升高.结论SjC23 DNA疫苗通过基因枪免疫也能产生部分免疫保护作用,但明显低于肌肉注射的方法.     

日本血吸虫多价DNA疫苗免疫保护作用的研究

目的研究日本血吸虫多价DNA疫苗诱导BALB/c小鼠的免疫保护作用.方法65只BALB/c雌性小鼠随机分为5组,各组小鼠分别肌肉注射纯化的质粒DNA pcDNA3.1(对照组)、pcDNA3.1-TPI(TPI)、pcDNA3.1-(CDR3)6[(CDR3)6]、pcDNA3.1-TPI-linker-(CDR3)6(TLC)和pcDNA3.1-(CDR3)6-linker-TPI(CLT),每鼠100μg.每隔2周加强免疫1次,共3次.末次免疫后4周每鼠经腹部皮肤攻击感染(45±1)条日本血吸虫尾蚴,45 d后剖杀,计数成虫及肝脏虫卵数.首次免疫前2天及感染前2天经尾静脉采血检测IgG抗体水平、抗体亚类IgG1、IgG2a.末次免疫后2周取小鼠脾脏制备单个脾细胞,检测细胞因子IL-2、IL-4、IFN-γ的水平.结果TPI、TLC组和CLT组小鼠血清都检测到特异性IgG抗体,抗体亚类IgG2a/IgG1的比值分别为1.71、2.71和4.70,而其他两组则未检测到特异性IgG抗体;TPI、TLC组和CLT组小鼠的IL-2、IFN-γ较对照组均有不同程度的升高,IL-4则无明显差异.多价DNA疫苗TLC组和CLT组减虫率分别达到37.30%、39.09%,减卵率分别为41.61%、49.54%,均显著高于TPI组和(CDR3)6组(P均＜0.05).结论多价DNA疫苗相对于单价DNA疫苗能诱导小鼠产生较高的免疫保护作用,且诱导宿主产生较高的以Th1为主的免疫应答.     

日本血吸虫中国大陆株TPI DNA疫苗诱导BALB/c小鼠保护性免疫力的研究

目的研究日本血吸虫中国大陆株磷酸丙糖异构酶(SjCTPI)DNA疫苗诱导BALB/c小鼠的保护性免疫作用.方法将39只BALB/c小鼠随机分成3组A组(对照组),肌注pcDNA3.1质粒DNA 100μg/鼠;B组(实验组),每鼠肌注pcDNA3.1-SjCTPI 100μg;C组(加强组),每鼠肌注pcDNA3.1-SjCTPI 100μg及pcDNA3.1-P35和pcDNA3.1-P40的混合物100μg.每两周免疫1次,共计3次.末次免疫4周后,每鼠用45条尾蚴进行攻击,45 d后剖杀小鼠,计数成虫以及每鼠肝组织内虫卵数;于攻击前每组各取两只小鼠采用51Cr释放法测定SjCTPI介导的特异性细胞毒作用;ELISA检测攻击前后的抗TPI的抗体水平.结果用ELISA检测抗TPI抗体的结果表明,攻击前A组的10份血清均为阴性,B组5/10份血清出现弱阳性,C组也有6/10份血清出现弱阳性反应.51Cr释放法测定细胞毒活性结果表明,A、B、C组细胞毒活性分别为9.1%、27.6%和54.4%.与对照组相比,实验组的减虫率、减卵率分别为30.2%、52.9%;加强组的减虫率、减卵率分别为32.7%、47.0%.结论进一步证实了SjCTPI DNA疫苗作为抗血吸虫病核酸疫苗的潜力.     

日本血吸虫中国大陆株TPI DNA疫苗诱导BALB／c小鼠保护性免疫力的研究

目的 研究日本血吸虫中国大陆株磷酸丙糖异构酶（SjCTPI)DNA疫苗诱导BALB/c小鼠的保护性免疫作用。方法 将39只BALB/c小鼠随机分成3组：A组（对照组），肌注pcDNA3.1质粒DNA100μg/鼠；B组（实验组），每鼠肌注pcDNA3.1-SjCTPI100μg;C组（加强组），每鼠肌注pcDNA3.1-SjCTPI100μg及pcDNA3.1-P35和pcDNA3.1-P40的混合物100μg。每两周免疫1次，共计3次，末次免疫4周后，每鼠用45条尾蚴进行攻击，45d后剖杀小鼠，计数成虫以及每鼠肝组织内虫卵数；于攻击前每组各取两只小鼠采用^51Cr释放法测定SjCTPI介导的特异性细胞毒作用。ELISA检测攻击前后的抗TPI的抗体水平。结果 用ELISA检测抗TPI抗体的结果表明，攻击前A组的10份血清均为阴性，B组5/10份血清出现弱阳性，C组也有6/10份血清出现弱阳性反应。^51Cr释放法测定细胞毒活性结果表明，A、B、C组细胞毒活性分别为9.1%、27.6%和54.4%。与对照组相比，实验组的减虫率、减卵率分别为30.2%、52.9%，加强组的减虫率、减卵率分别为32.7%、47.0%。结论 进一步证实了SjCTPI DNA疫苗作为抗血吸虫病核酸疫苗的潜力。     

CpG免疫刺激序列增强日本血吸虫TPIDNA疫苗免疫保护作用的研究

目的研究CpG免疫刺激序列对日本血吸虫中国大陆株磷酸丙糖异构酶DNA疫苗诱导BALB/c小鼠免疫保护的增强作用.方法设计合成一对含6个免疫刺激序列的寡脱氧核苷酸,克隆到真核表达载体pcDNA3.1第5314碱基Ssp I酶切位点,改建为pcDNA3.1-CpG.再将SjCTPIDNA片段克隆到pcDNA3.1-CpG的多克隆位点区,构建为pcDNA3.1-TPI/CpG.56只BALB/c雌性小鼠随机分为4组,各组小鼠分别肌肉注射pcDNA3.1、pcDNA3.1-TPI、pcDNA3.1-CpG、pcD-NA3.1-CpG/TPI质粒DNA,每鼠100 μg.每隔2周加强免疫1次,共3次.末次免疫后4周每鼠经腹部皮肤攻击感染(45士1)条日本血吸虫尾蚴,45 d后剖杀,计数成虫及肝脏虫卵数.首次免疫前2d及攻击感染前2 d经尾静脉采血检测抗体水平、抗体亚类IgG1、IgG2a.末次免疫后2周制备脾细胞,检测细胞因子IL-2、IL-4、IFN-γ的水平.结果 pcDNA3.1-TPI组和pcDNA3.1-TPI/CpG组小鼠血清都检测到特异性IgG抗体,抗体亚类IgG2./IgG1的比值分别为1.76和2.67,而其他两组则未检测到特异性IgG抗体;pcDNA3.1-TPI组和pcDNA3.1-TPI/CpG组小鼠的IL-2、IFN-γ较对照组均有不同程度的升高,IL-4无明显差异;pcDNA3.1-TPI组和pcDNA3.1-TPI/CpG组分别获得了25.98%和34.54%的减虫率,后者高于前者,并分别获得了27.68%和29.50%的减卵率.结论 CpG免疫刺激序列似能增强DNA疫苗在小鼠体内诱导的Th1型免疫应答,并提高DNA疫苗的免疫保护作用.     

日本血吸虫TPI DNA疫苗基因密码子优化增强免疫保护作用的研究

目的　 研究日本血吸虫中国大陆株磷酸丙糖异构酶基因（TPI基因）密码子优化后的DNA疫苗增强免疫保护作用的效果。 方法　 60只BALB/c雌性小鼠随机均分为A（pcDNA3.1空质粒对照组）、B（pcDNA3.1-TPI组）、C （pcDNA3.1-TPI-mHSP70组）、D（pcDNA3.1-TPI.opt组）、E（pcDNA3.1-TPI.opt-mHSP70组）等5组。每鼠肌肉注射相应的纯化质粒DNA 100 μg,每隔3周免疫1次,共3次。末次免疫后4周,每鼠经腹部皮肤攻击感染日本血吸虫尾蚴（40±1）条,42 d后剖杀,计数成虫及肝脏虫卵数。首次免疫前2 d及感染前2 d经尾静脉采血,检测IgG及IgG₁、IgG_2a的水平。攻击感染前2 d取脾脏,制备单个脾细胞,用流式细胞仪检测白细胞介素2（IL-2）、IL-4、IL-5、γ干扰素（IFN-γ）及肿瘤坏死因子（TNF）的水平。 结果　 B、C、D、E组小鼠血清均检测到特异性IgG及IgG_2a与IgG₁抗体,IgG_2a/IgG₁的比值分别为1.73、2.06、2.44、3.09。D、E组的IL-2、IFN-γ、TNF含量较B、C组均有不同程度地升高。D、E组减虫率分别为36.03%、39.03%,减卵率分别为41.71%、46.85%,均显著高于B、C组（P<0.01）。 结论 TPI基因密码子优化后的DNA疫苗相对于未优化TPI DNA疫苗能诱导小鼠产生较高的免疫保护作用,且诱导宿主产生较强的,及以Th1为主的免疫应答。     

血吸虫硫氧还蛋白免疫原性研究Ⅱ日本血吸虫硫氧还蛋白DNA疫苗小鼠保护性免疫研究

目的观察日本血吸虫(大陆株)硫氧还蛋白DNA疫苗(pcDNA3-SjcTrx)在小鼠诱导抗血吸虫感染的免疫保护作用。方法制备pcDNA3-SjcTrx重组质粒,将30只雌性C57BL/6小鼠随机分为3组,每组10只:pcDNA3-SjcTrx核酸疫苗免疫组、pcDNA3空质粒对照组和攻击感染对照组。核酸疫苗免疫组每只小鼠经股四头肌注射100μg核酸疫苗,共注射3次,间隔2周。空质粒对照组每只小鼠在相应时间经股四头肌注射100μgpcDNA3空质粒,感染对照组则不注射任何质粒。于末次免疫后3周,每只小鼠经腹部感染(30±1)条日本血吸虫尾蚴。小鼠于攻击感染后42天剖杀,门脉灌注收集成虫,计数成虫数和肝内虫卵数。分别在免疫前、攻击感染前和小鼠剖杀前采血并分离血清,用ELISA检测血清中特异性IgG抗体。另取6只雌性C57BL/6小鼠经股四头肌注射核酸疫苗,分别于注射后24、48小时和72小时取肌肉注射部位局部组织制备冰冻切片,用免疫酶染色试验(IEST)检测注射局部组织抗原的表达情况,以注射pcDNA3空质粒者为对照。结果IEST结果表明该DNA疫苗在小鼠肌肉组织内表达,ELISA检测表明DNA疫苗免疫小鼠后产生明显的抗体(IgG)免疫应答,并诱导出对攻击感染的45.7%的减虫率和41.4%的肝组织减卵率(P<0.05)。结论日本血吸虫(大陆株)硫氧还蛋白DNA疫苗具有较好的免疫原性,在小鼠诱导出明显的免疫保护作用,可作为疫苗候选分子作进一步的研究。     

日本血吸虫磷酸丙糖异构酶(TPI)DNA疫苗对肝脏虫卵肉芽肿调节作用的研究

目的 探讨日本血吸虫TPI DNA疫苗对血吸虫肝脏虫卵肉芽肿的免疫调节作用，以评价其作为抗病疫苗的潜能。方法 30头松江猪分为3组。A组(TPI组)每头猪于两侧臀部肌注500μg pcDNA3．1-SjCTPI质粒DNA；B组(TPI IL-12组)每头猪肌注500μg pcDNA3．1-sjCTPI DNA疫苗及500μg pcDNA3．1-P35和500μg pcDNA3．1-P40质粒DNA的混和物；C组(对照组)，每头猪肌注500μg pcDNA3．1质粒DNA。于第0、3、6周共免疫3次。末次免疫后30d，每头猪采用背部贴片法攻击感染600条日本血吸虫尾蚴，攻击后45d剖杀实验猪，取肝脏，于同一部位切取1．5cm^3的肝组织。按常规法制备石蜡切片。切片在NYD-1000型图像分析系统下观察，测定所有肉芽肿的面积和直径，并比较。结果 与对照组相比TPI组及TPI IL-12组肉芽肿内炎性细胞明显减少。肉芽肿平均面积较对照组分别减少41．95％和42．50％，肉芽肿直径分别减少25．14％和25．66％。结论 SjCTPI DNA疫苗具有下调血吸虫肝脏肉芽肿的作用，可望成为一种血吸虫病抗病疫苗。     

日本血吸虫调宁蛋白样蛋白P14基因DNA疫苗对小鼠免疫保护作用研究

摘 要：目的 研究日本血吸虫调宁蛋白样蛋白P14基因DNA疫苗对小鼠免疫保护作用。 方法 制备无内毒素DNA疫苗,用于免疫小鼠。将雌性BALB/c小鼠随机分为4组,每组10只。生理盐水组给予100 μl/鼠/次;空质粒组100 μg/鼠/次、pcDNA3.1(+)-SjP14组、pcDNA3.1(+)-SjP14 + pcDNA3.1(+)-SjGST组分别经肌肉给予100 μg/鼠/次;同上每2周免疫一次,共3次。末次免疫后2周,经腹部皮肤感染日本血吸虫尾蚴（30±1）条/鼠。尾蚴攻击6周后解剖小鼠,收集成虫和血清,计算减虫率并检测血清IgG1、IgG2a及总IgG;同时留取肝脏,部分消化后在显微镜下行虫卵计数,计算减卵率,部分肝脏用于组织病理学分析（HE染色法）,观察肝细胞变化及肉芽肿情况。结果 与NS对照组比较,pcDNA3.1(+)-Sj P14p核酸疫苗组减虫率和减卵率分别达到45.1%（P<0.05）和62.0%（P<0.001）;SjP14核酸疫苗与SjGST核酸疫苗联合免疫组减虫率和减卵率分别提高至56.3%（P<0.01）和73.9%（P<0.001）;SjGST和SjP14 组减虫率大于SjP14疫苗组,但两组之间差异无显著性（P＞0.05）; SjGST和SjP14组减卵率明显大于SjP14疫苗组, 两组之间差异有显著性（P<0.001）。SjGST和SjP14组与SjP14组血清IgG1、IgG2a及总IgG水平在免疫6周、12周后均较对照组显著提高（P<0.01）,但SjGST和SjP14组与SjP14之间差异无显著性（P＞0.05）。肝组织切片镜下显示,pcDNA3.1(+)-SjP14组肝脏病变较生理盐水组和空质粒组明显减轻,pcDNA3.1(+)-SjP14 + pcDNA3.1(+)-SjGST组肝脏损伤程度最轻,虫卵肉芽肿周围炎症反应轻,肉芽肿面积较小。结论 pcDNA3.1(+)-Sj P14核酸疫苗有一定程度的抗血吸虫感染作用,pcDNA3.1(+)-SjP14与pcDNA3.1(+)-SjGST疫苗联合免疫能增强小鼠对血吸虫感染的保护。     

日本血吸虫P14基因纳米微球-DNA疫苗的免疫保护作用

目的 探讨日本血吸虫碱性调宁蛋白样蛋白P14基因的重组真核表达载体(pcDNA3.1(+)-SjP14)对小鼠血吸虫感染的免疫保护作用.方法 制备无内毒素pcDNA3.1 (+)-SjP14及纳米微球DNA疫苗,将雌性BALB/c小鼠随机分为3组,每组10只,分别为生理盐水对照组、pcDNA3.1(+)-Sj P14组及纳米微球-DNA疫苗组.各组小鼠分别肌肉注射生理盐水和pcDNA3.1(+)-Sj P14、纳米微球DNA疫苗100μg/鼠·次,每2周免疫一次,共3次.末次免疫后2周,尾蚴攻击,6w后处死小鼠,收集成虫,计算减虫率;同时留取肝脏,部分消化后在显微镜下行虫卵计数,计算减卵率.结果 pcDNA3.1(+)-Sj P14p核酸疫苗能明显提高小鼠的抗血吸虫感染能力,减虫率和减卵率分别达到44.6％和61.6％;纳米微球-DNA疫苗免疫能进一步提高小鼠抗血吸虫感染作用,减虫率和减卵率分别提高至56.2％和73.5％.结论 pcDNA3.1(+)-Sj P14核酸疫苗有一定程度的抗血吸虫感染作用,pcDNA3.1(+)-SjP14-纳米微球复合物疫苗免疫能增强小鼠对血吸虫感染的免疫保护作用.     

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.     

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.     

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.     

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.     

High prevalence of distal sensory polyneuropathy in antiretroviral-treated and untreated people with HIV in Tanzania

Objectives To describe the prevalence of distal sensory polyneuropathy (DSP), a complication of both advanced HIV disease and of antiretroviral therapy (ART), amongst Tanzanians with HIV, on and off ART (including stavudine) with CD4 counts above and below 200 cells/μl. Methods We recruited participants attending ART clinic into four groups: >6 months ART exposure and (i) CD4 < 200 cells/μl or (ii) CD4 > 200 cells/μl (ART/CD4 < 200 and ART/CD4 > 200, respectively); ART‐naïve and (iii) CD4 < 200 cells/μl or iv)CD4 > 200 cells/μl (noART/CD4 < 200 and noART/CD4 > 200, respectively). Primary outcome was DSP, as defined by presence of at least one symptom and one sign. Results Of 326 evaluable participants, 81 (32 men, median age 38 years, median CD4 142 cells/μl) were enrolled in the ART/CD4 < 200 group, 78 (17 men, median age 37 years, median CD4 345 cells/μl) in ART/CD4 > 200, 81 (30 men, median age 37 years, median CD4 128 cells/μl) in noART/CD4 < 200 and 86 (22 men, median age 33 years, median CD4 446 cells/μl) in noART/CD4 > 200. Numbness was the most commonly reported symptom. DSP prevalence ranged from 43.2% in ART/CD4 < 200 to 20.9% in noART/CD4 > 200. DSP was more common among men (adjusted odds ratio [aOR] 1.9, 95% confidence interval [CI] 1.2–3.3) and older participants (aOR 2.7, 95% CI 1.1–6.2 for age 40 + vs. <30 years). Conclusion Distal sensory polyneuropathy is common amongst those attending this clinic, even those with no ART exposure and a CD4 count above 200 cells/μl. Stavudine and didanosine expose HIV‐infected patients to an additional avoidable risk of DSP. Access to non‐neurotoxic ART regimes as well as earlier HIV diagnosis and initiation of ART is needed.