喀什地区两起流行性出血热实验室检测和溯源调查

目的 查明新疆喀什地区两起（莎车县、叶城县各一起）不明原因疑似出血热是否为流行性出血热汉坦病毒感染及其感染方式。方法 现场调查患者居住场所环境及饮食卫生,在患者居住场所及周边环境捕鼠并采集标本,采用酶联免疫吸附试验和实时荧光定量PCR试验检测标本。结果 莎车县和叶城县各一例疑似流行性出血热患者血清汉坦病毒IgM抗体均呈阳性,其中莎车县病例抗体滴度1∶6 400,叶城县抗体滴度1∶12 800;疑似流行性出血热汉坦病毒特异核酸检测,其中莎车县阴性,叶城县阳性（FAM通道Ct值阴性,HEX通道Ct值36.809）;患者居住场所及周边环境共捕获啮齿动物小家鼠4只、褐家鼠24只,其中小家鼠流行性出血热汉坦病毒特异核酸检测均阴性,褐家鼠阳性6份、阳性率25.00%。结论 喀什地区两起疑似出血热均为流行性出血热汉坦病毒（汉城型）感染,感染方式为通过食用被褐家鼠粪便、尿液污染的食物后经消化道感染流行性出血热汉坦病毒（汉城型）。     

白城市洮北区首例肾综合征出血热流行病学调查

目的：核实诊断肾综合征出血热(HFRS)临床病例、疫情，确认疫源地，查找疫源动物。方法：运用了流行病学横断面调查方法。结果：在疫点内调查30人，其中5人HFRS IgG抗体阳性，隐性感染率为16．67％。捕到褐家鼠19只，其中2只HFRS病毒抗原阳性，带毒率为10．53％。结论证实白城市HRFS 临床病例及疫情存在，可以确认家鼠型HFRS疫源地。     

三种ELISA法检测大鼠血清肾综合征出血热病毒特异性抗体的研究

肾综合征出血热(HFRS)是由布尼亚病毒科汉坦病毒属引起的自然疫源性疾病。本病的传染源除黑线姬鼠、褐家鼠、欧洲棕背外,能作为家鼠型或实验动物型HFRS传染源的动物,还有实验大白鼠。近十年来世界各地相继暴发了由实验大鼠引起的HFRS流行,日益引起了人们的高度重视,迫切需要一些快速、简便、敏感、特异的检测手段以用于野生及实验动物的HFRS病毒流行病学监测及检疫,以预防野鼠型、家鼠型及实验室型HFRS的发生。虽然间接免疫荧光试验(IFA)一直是主要可靠的血清学诊断手段,但由于需用昂贵的仪器,并且判定结果有一定的主观性,降低了它的应用价值。我们建立了三种检测实验大鼠体内HFRS病毒抗体的ELISA方法,并对这些方法的敏感性。 (一)取代试验:1.用相应包被液或稀释液代替反应系统中一种或二种成分,结果均为阴性(见表1)。2.以JEV抗原和抗体,HSV抗原和抗体分别取代HFRS病毒抗原和A35McAb、Mark-1 McAb及免抗HFRS病毒免疫血清r-G。以酶标免抗人IgG取代酶标Mark-l,酶标H7 McAb和PAP复合物,羊抗人IgG取代桥抗免抗大鼠血清,结果均为阴性(见表2)。     

福建省鼠间肾综合征出血热流行病学调查

目的掌握福建省鼠间肾综合征出血热(HFRS)流行情况和疫源地分布,为控制HFRS的流行提供参考。方法根据福建省啮齿动物地理区划以及肾综合征出血热病例分布状况,选择18个县、市作为调查地区,用笼夜法捕鼠采集血清标本,用双抗原夹心ELISA法检测血清中抗汉坦病毒(Hantan virus,HV)抗体,分析鼠间HFRS流行情况。结果各个地区HV宿主动物种类分布不均衡,但以褐家鼠为主要宿主动物,其次为黄胸鼠,野外鼠密度较低。褐家鼠抗体阳性率最高,达到9.89%。各个地理区划中II区即闽东山地抗体阳性率最高,为9.32%,而III区即闽中、闽西南山地抗体阳性率最低,为2.74%。调查点中泰宁、周宁、福鼎、厦门、福清、莆田、平潭等地鼠血清HV抗体阳性率均超过10%。而上杭、漳平、漳浦、安溪等地鼠血清均未检出HV抗体。结论福建省HFRS的疫源地以家鼠型疫区为主,降低家鼠尤其是褐家鼠密度,可控制本病在福建的扩大流行。     

云南省2001～2002年肾综合征出血热监测研究

目的为掌握云南省肾综合征出血热流行病学特点,提供防治参考,对人间和鼠间疫情进行了监测.方法收集全省本病疫情资料,并在监测县采集人血清以及鼠肺脏和鼠血清作汉坦病毒抗原和抗体检查.结果 2001～2002年全省共报告本病102例,死亡3例,年发病率为0.12/10万,病死率为2.94%.主要发病地区为红河州、昆明市、楚雄州.疫区人群隐性感染率为4.19%.2002年在泸西、寻甸和永胜监测点捕获鼠类9种891只,居民区以褐家鼠和黄胸鼠为优势鼠种,野外以高山姬鼠为优势种;鼠间汉坦病毒带毒率为3.65%,带病毒鼠种为褐家鼠、黄胸鼠、小家鼠和高山姬鼠.2001年在大理市野外捕鼠12种140只,大绒鼠为优势种,带毒鼠为大绒鼠、大足鼠、黄胸鼠、社鼠和短尾鼠句.结论监测区内存在有以褐家鼠和黄胸鼠为主要宿主动物的家鼠型疫源地,也存在着以高山姬鼠和大绒鼠为主的野鼠型疫源地.发病率上升与较高的鼠密度和鼠间感染率有关.应采取以灭家鼠和接种家鼠型或两型混合疫苗为主的防治措施.     

柏氏禽刺螨传播家鼠型汉坦病毒的研究

目的：研究柏氏禽刺螨在传播家鼠型肾综合征出血热病毒中的作用。方法：每罐放柏氏禽刺螨100只，于室温为23±1℃环境下饥饿4d，分别叮刺感染汉坦病毒（Hantaanvirus）或浙45株（Z45）大鼠乳小鼠2只或感染汉城病毒（Seoulvirus）UR株大鼠乳小鼠2只各12h，为感染螨，用感染后d15的螨叮刺正常大鼠乳小鼠9只，d30取感染鼠的肺和血清，用间接免疫荧光测试。于d15和d23取感染螨用聚合酶链反应测试其体内病毒RNA。结果：柏氏禽刺螨分别叮刺感染汉坦病毒Z45株和感染汉城病毒UR株大鼠乳小鼠后, 再分别叮刺正常大鼠乳小鼠5 只和4 只。IFAT 检测结果汉坦病毒Z45 株组的鼠全部阳性, 汉城病毒UR 株组的4 只鼠中3 只阳性, 其抗体滴度, 前者为10- 40, 后者为20- 40, 病毒在螨体内可保存22 d 以上。阻断试验显示, 病毒免疫血清1∶20 作用肺切片标本后, 部分阻断荧光抗体反应, 与未经稀释的血清作用后全部阻断特异性荧光反应。对照组正常鼠肺片和血清经IFA T 检测均阴性。结论: 柏氏禽刺螨可作为家鼠型肾综合征出血热的传播媒介和贮存宿主。     

猪乙脑病毒IgG抗体间接免疫荧光检测方法的建立与初步应用

目的建立检测猪血清中流行性乙型脑炎病毒(JEV)IgG抗体的间接免疫荧光试验(IFA)。方法用乙脑病毒感染C6/36白蚊伊蚊细胞制备抗原片,建立检测猪血清中JEV-IgG抗体的IFA方法,并用于猪乙脑血清流行病学调查。结果成功建立特异的猪JEV-IgG的IFA检测方法,并对50份母猪、55份仔猪和65份屠宰猪的血清进行检测。乙脑血清抗体效价≥1:200的分别为88.00%,10.91%和24.62%。结论建立的IFA方法能较好的用于猪血清乙脑流行病学调查。     

汉坦病毒重组核蛋白抗体用于鼠肺组织病毒抗原的检测

目的纯化汉坦病毒(HV)SEO型L99株重组核蛋白(rNP),制备抗体用于鼠肺组织HV抗原的检测。方法重组菌株E.coliBL21(DE3)/pET32a-L99S经IPTG诱导表达rNP,采用Ni亲和层析方法纯化,纯化蛋白免疫日本大耳白兔,获取抗血清用于间接免疫荧光法(IFA)检测鼠肺组织的HV抗原;并与病人混合血清作为一抗的IFA结果进行比较,部分标本进一步用RT-PCR法进行验证。结果rNP表达量占菌体总蛋白的52.2%,纯化蛋白的总回收率为22.9%;纯化蛋白Western blot分析为单带,纯度达95%以上,用其制备多克隆抗体滴度达1∶512000。IFA检测59份鼠肺标本,用兔rNP抗体和病人混合血清作为一抗其阳性率分别为23.7%和16.9%;两者结果不一致的6份鼠肺经RT-PCR检测HV核酸,均与兔rNP抗体的IFA结果相一致。结论采用rNP抗体进行IFA检测,可显著地提高鼠类HV感染监测的准确性,值得推广使用。     

应用间接免疫荧光试验诊断丝虫病

作者等在新加坡应用马来丝虫微丝蚴超声粉碎抗原间接免疫荧光试验(IFA)检测微丝蚴血症阴性的疑似丝虫病患者血清中丝虫抗体。抗原制备及IFA分别参照Singh(1977)和Kane(1971)方法进行。以微丝蚴碎片边缘及表膜上出现黄绿色荧光为阳性反应。在324份有各种丝虫病症状体征而无微丝蚴血症患者的血清中仅28%(90/324)为IFA阳性。抗体滴度自1:4至1:4096。绝大部分血清滴度在1:4至1:256之间,其中1:4、     

云南省恙虫病与肾综合征出血热混合感染的诊断及分析

目的 调查云南省不明原因发热患者中是否存在恙虫病与肾综合征出血热混合感染病例.方法 采用间接免疫荧光法和胶体金法检测不明原因发热患者血清中的恙虫病东方体(OT)抗体及肾综合征出血热(HFRS)病毒抗体,用巢式PCR扩增患者血液中的OT 基因sta56片断与汉坦病毒(HV)基因L片断.结果 79例患者中,实验室确诊HFRS 5例,恙虫病3例,HFRS与恙虫病混合感染4例;巢式PCR检测到HV L片断核酸2例(DLR2和DLR6), 序列分析两者间的同源性100%,DLR2与老挝汉城病毒L0199株核苷酸序列同源性最高为94.6%, 与中国汉城病毒L99株的同源性80.5%, 与云南汉坦病毒YN06-256的同源性69.8%.结论 首次证实云南省存在恙虫病与肾综合征出血热混合感染病例,其HFRS病原体遗传进化关系与老挝汉城病毒L0199株密切相关.     

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.     

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.     

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.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.