SS／GHRH－GH－IGF－Ⅰ轴与糖尿病

ＳＳ／ＧＨＲＨ－ＧＨ－ＩＧＦ－Ⅰ轴与糖尿病廖二元，许樟荣胰岛素样生长因子（ｉｎｓｕｌｉｎ－ｌｉｋｅｇｒｏｗｔｈｆａｃｔｏｒｓ，ＩＧＦｓ）为一组分子量约７５００Ｄａ的蛋白质激素，分ＩＧＦ－Ⅰ和ＩＧＦ－Ⅱ两种，现已查明，ＩＧＦ－Ⅰ即生长介素Ｃ。垂体分泌的...     

重组质粒pGEX—6P—1／Sj—FABPc的构建及在大肠杆菌中的表达

目的 构建基因工程菌株、获得重组蛋白Ｓｊ－ＦＡＢＰｃ（日本血吸虫脂肪酸结合蛋白）。方法 用ＰＣＲ法从日本血吸虫ｃＤＮＡ文库中扩增Ｓｊ－ＦＡＢＰｃ基因片段，再将该片段重组于ｐＧＥＭ－Ｔ中并进行ＤＮＡ测序鉴定，经酶切后将目的片段构建成重组质粒ｐＧＥＸ－６Ｐ－１／Ｓｊ－ＦＡＢＰｃ，转化于大肠杆菌ＢＬ２１，ＩＰＴＧ诱导表达。用Ｇｌｕｔａｔｈｉｏｎｅ Ｓｅｐｈａｒｏｓｅ＾ＴＭ ４Ｂ亲和层析柱对表达产物进行纯     

日本血吸虫26kDa谷胱甘肽S-转移酶基因真核表达载体的构建及序列测定

目的 扩增日本血吸虫２６ｋＤａ谷胱甘肽Ｓ－转移酶（Ｓｊ２６ＧＳＴ）基因片段，构建其重组真核表达载体，以研制ＳｊＧＳＴ核酸疫苗。方法 根据已知基因序列设计一对引物，运用ＲＴ－ＰＣＲ技术扩增Ｓｊ２６ＧＳＴ目的基因片段，将其克隆到ｐｃＤＮＡ３质粒中，并经酶切、ＰＣＲ鉴定，测序证实。结果 获得ＧＳＴ－ｐｃＤＮＡ３重组克隆。结论：本实验获得Ｓｊ２６ＧＳＴ重组克隆，为进一步研制核酸疫苗创造了条件。     

恶性疟原虫海南FCC1/HN株环子孢子蛋白(CSP)基因的克隆与表达

目的　为疟疾疫苗的研制提供靶抗原。方法　根据恶性疟原虫ＩＭＴＭ２２ 株７Ｇ８ 克隆环子孢子蛋白基因编码区序列, 设计一对引物, 采用ＰＣＲ技术从恶性疟原虫ＦＣＣ１／ＨＮ株基因组ＤＮＡ中特异扩增ＣＳＰ基因的Ⅰ区、中央重复区和Ⅱ区片段, 全长１－０８ｋｂ; 纯化扩增产物用ＨｉｎｄⅢ和ＢａｍＨ Ⅰ双酶切后, 定向克隆入ｐｃＤＮＡ３ 载体, 转化大肠杆菌ＴＧ１ 株, 重组克隆经筛选后, 用ＰＣＲ 扩增和ＨｉｎｄⅢ＋ ＢａｍＨ Ⅰ双酶切进行鉴定： 用磷酸钙贴壁细胞转化法将重组质粒ｐｃＤＮＡ—ＣＳＰ导入ＨｅＬａ细胞, 用Ｇ４１８ 筛选出稳定分泌ＣＳＰ抗原的阳性细胞克隆; 将阳性细胞克隆系扩大培养并用Ｇ４１８ 加压, 以表达重组ＣＳＰ, 分离细胞培养上清和培养细胞, 进行ＳＤＳ ＰＡＧＥ分析。结果　（１） 从ＦＣＣ１／ＨＮ株基因组ＤＮＡ中特异扩增出ＣＳＰ基因Ⅰ区, 中央重复区和Ⅱ区编码序列;（２） 将扩增的目的片段正向插入ｐｃＤＮＡ３ＨｉｎｄⅢ和ＢａｍＨⅠ位点; （３） 在人宫颈癌细胞系ＨｅＬａ 细胞中表达重组ＣＳＰ抗原, 其分子量为３８－３ｋＤａ, 蛋白扫描分析表达量占细胞培养上清液中蛋白总含量的１５－７７％ 。结论　成功构建真核表达系统ｐｃＤＮＡ３     

日本血吸虫基因工程BCG多价疫苗的研究

用日本血吸虫（Ｓｊ）成虫提取的总ＲＮＡ逆转录合成ｃＤＮＡ第一链，并设计合成引物，经ＰＣＲ扩增，扩增出２６ｋＤａ的编码区基因，并进行了测序，结果表明Ｓｊ中国大陆株（ＳｊＣ）、Ｓｊ菲律滨株（ＳｊＰ）２６ｋＤａ ＧＳＴ基因核苷酸同源性９９．８％。然后，构建了大肠杆菌－分枝杆菌穿梭表达质粒ＰＢＣＧ－Ｓｊ２６Ⅰ￣Ⅳ。再将其依次分别转化大肠杆菌、耻垢分枝杆菌和ＢＣＧ中，筛选出重组耻垢分枝杆菌疫苗和重组ＢＣＧ多     

含日本血吸虫脂肪酸结合蛋白（SjFABPc）基因DNA重组质粒 …

目的 观察裸ＤＮＡ重组质粒ｐＣＤ－ＳｊＦＡＢＰｃ在体外转染ＨｅｐＧ２细胞以及质粒ＤＮＡ免疫鼠肌组织中的表达，为进一步应用该质粒进行ＤＮＡ免疫实验奠定基础。方法 用脂质体介导ＤＮＡ转染法将ｐＣＤ－ＳｊＦＡＢＰｃ转染贴壁细胞ＨｅｐＧ２，Ｇ４１８加压筛选获得阳性克隆细胞并传代培养，ＳＤＳ－ＰＡＧＥ及Ｗｅｓｔｅｒｎ－ｂｌｏｔ鉴定目的基因在ＨｅｐＧ２细胞中的表达，将ｐＣＤ－ＳｊＦＡＢＰｃ质粒肌注免疫ＢＡＬＢ     

反义c-mycRNA的表达对大鼠血管平滑肌细胞c-mycmRNA和c-Myc蛋白水平的影响

为探讨反义ｃ－ｍｙｃ逆转录病毒载体是否影响血管平滑肌细胞表达ｃ－ｍｙｃ ｍＲＮＡ和ｃ－Ｍｙｃ蛋白水平，用基因重组方法，将第二外显子及其两则部分内含子共１．５３ｋｂ序列的ｃ－ｍｙｃ基因片段反向连接于Ｐ＾ＬＸＳＮ的５′长末端重复序列下游，通过Ｌｉｐｏｆｅｃｔｉｎ将反义ｃ－ｍｙｃ逆转录病毒导入ＰＡ３１７包装细胞系，并经Ｇ４１８筛选获得抗性克隆。反义－ｃ－ｍｙｃ逆转录病毒上清液转染的平滑肌细胞经ＤＮＡ印迹     

日本血吸虫中国大陆株成虫谷胱甘肽S—转移酶cDNA克隆及其核苷酸？…

日本血吸虫成虫２６ｋＤ谷胱甘肽Ｓ－转移酶是成虫代谢过程中一个关键酶。为研究制备日本血吸虫重组疫苗，根据日本血吸虫菲律宾株成虫Ｓｊ２６ｋＤＧＳＴ完整编码区序列设计引物，以日本血吸虫中国大陆株成虫２６ｋＤＧＳＴｍＲＮＡ模板，采用逆转 聚合酶链反应技术，扩增了其２６ｋＤＧＳＴ完整编码区ｃＤＮＡ将ｃＤＮＡ重组于ｐＢｌｕｅｓｃｒｉｐｔＳＫ质粒上，转化大肠杆菌，重组体经限制酶解图谱鉴定，用双脱氧链末端终止法对     

谷氨酸脱羧酶表达型质粒的构建

采用逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）克隆了谷氨酸脱羧酶基因的ｃＤＮＡ，双脱氧末端终止法对其全部核苷酸序列予以确定后，将此编码５８５个氨基酸、全长１７５８ｂｐ的ｃＤＮＡ重组入ｐＧＥＸ表达型载体中，通过原位杂交筛选阳性重组质粒，并通过序列分析证实重组质粒接口处读码框架无误，为谷氨酸脱羧酶重组基因的表达及表达产物在临床诊断中的应用奠定了基础。     

基因重组人粒细胞集落刺激因子防治肺癌患者化疗反应

采用随机分组、自身交叉对比的方法，观察了基因重组人粒细胞集落刺激因子（Ｒｅｃｏｍｂｉｎａｎｔｈｕｍａｎｇｒａｎｕｌｏｃｙｔｅｃｏｌｏｎｙ－ｓｔｉｍｕｌａｔｉｎｇｆａｃｔｏｒ，ｒｈＧ－ＣＳＦ）对肺癌患者化疗（卡铂＋鬼臼乙叉甙，即ＣＥ化疗方案）所致白细胞和中性粒细胞减少的防治作用及毒副反应。全组共２２例，随机分成Ａ、Ｂ两组（各１１例）。Ａ组第一周期化疗后加用ｒｈＧ－ＣＳＦ，第二周期单用化疗；Ｂ组第一周期单用化疗，第二周期化疗后加用ｒｈＧ－ＣＳＦ。结果表明，ｒｈＧ－ＣＳＦ可以明显减轻化疗过程中白细胞和中性粒细胞下降的程度及其发生率，缩短白细胞和中性粒细胞降至正常值以下的持续时间，促进其早日恢复，使化疗可以如期进行。ｒｈＧ－ＣＳＦ副反应轻微，安全可靠。     

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.