生物安全三级实验室人员管理相关问题的探讨

在生物安全三级(BSL-3)实验室日常管理工作中,实验人员的管理是实验室安全运转的关键,可保证生物安全管理体系文件的准确执行,减少安全隐患.我们总结了数年来管理BSL-3实验室的经验和教训,对人员管理有关问题提出想法和建议,以供参考.     

上海市（复旦大学附属）公共卫生临床中心生物安全三级实验室

上海市(复旦大学附属)公共卫生临床中心生物安全三级实验室(简称中心BSL-3实验室)于2005年建造,同年通过国家建筑工程质量监督检验中心对实验室建筑结构的设计、实验室规范等各项指标的验收,获得<上海市公共卫生中心BSL-3实验室工程>检验报告.中心BSL-3实验室位于科研中心二楼北侧自成隔离区的建筑物内,周围最近的居民居住区在180m外.     

从新型冠状病毒疫情看生物安全三级实验室的应急能力及内涵建设

面对突发疫情,生物安全三级（biosafety level 3,BSL-3）实验室是否能及时启动并稳定、安全运行,为科研攻关提供安全、可靠的技术平台,是对实验室应急能力的考验。针对2019年12月爆发的2019冠状病毒病疫情,复旦大学BSL-3实验室在接到任务后,立即按照国家规定,启动实验室冠状病毒新实验活动申请流程。经国家合格评定认可委员会（China National Accreditation Service for Conformity Assessment,CNAS）和国家卫生委员会批准,实验室在开展实验活动一周内就从来自上海患者标本中分离到新型冠状病毒毒株,体现了平时实验室内涵建设的重要性。BSL-3实验室的内涵建设包括实验室风险评估及管理、不同人员针对性培训、设施和设备管理等生物安全管理体系的建设。本文将介绍复旦大学BSL-3实验室在应急能力及内涵建设方面的经验和体会,期望对BSL-3实验室的建设和管理提供参考。     

甘肃马鬃山盘羊春季卧息地的利用

2010年4～5月,在甘肃省马鬃山地区调查研究了盘羊的卧息地生境选择,共测得盘羊利用样方81个,对照样方81个。通过比较表明,马鬃山地区的盘羊夏季卧息地主要选择高海拔、阳坡、中坡位、坡度大、红砂植被、碎石地、植被密度低、远离道路和居民点、接近水源和隐蔽物、隐蔽程度好的区域。逻辑斯蒂回归分析表明,海拔(OR值1.614,95%CI1.248～4.419)、坡向(95%CI1.682～3.745)、坡位(95%CI0.839～3.156)、坡度(OR值1.705,95%CI1.227～2.369)、植被类型(95%CI0.682～2.732)、距水源距离(OR值0.180,95%CI0.041～0.787)、距道路距离(OR值1.628,95%CI2.598～3.941)、隐蔽级(OR值0.911,95%CI0.857～0.967)是盘羊春季卧息地选择的主导因子。     

医学病毒学生物安全三级动物实验室的建立

目前国内多个机构已建立了BSL-3实验室,这些实验室能够满足细胞水平和动物水平的病原微生物实验活动.医学病毒学生物安全三级动物实验室(ABSL-3)可分为小动物实验室和中(大)动物实验室.小动物实验室主要开展小鼠、大鼠、仓鼠等啮齿类动物的感染性实验活动;中(大)动物实验室主要开展小型猪、猴等实验.动物三级实验室特别是开展中(大)动物实验的高等级实验室,其实验室布局,设备设施,管理体系及流程会有很大的差别.本文从使用者的角度,通过所在机构医学病毒学生物安全三级动物实验室(ABSL-3)的建设与实践,对于筹备和建立以及使用过程中的所面对的问题以及应对,进行了提炼和归纳,分析和探讨,旨在为建设和使用医学病毒学高致病性生物安全三级动物实验室提供借鉴和参考.     

油田作业人员职业紧张状况调查及心理障碍的影响因素分析

目的:调查油田作业人员职业紧张状况,并分析心理障碍的影响因素。方法:通过分层整群抽样法纳入600例油田作业人员作为研究对象,采用职业紧张量表(OSI-R)以及症状自评量表(SCL-90)对油田作业人员的职业紧张程度以及心理障碍状况进行评估,并按照SCL-90评分将其分为研究组(SCL-90评分160分)214例与对照组(SCL-90评分≤160分)386例,采用多因素Logistic回归分析油田作业人员心理障碍的影响因素。结果:研究组年龄45岁、受教育程度为本科及以上、工龄20年人数占比均高于对照组(均P0.05)。研究组职业任务问卷(ORQ)、个体紧张反应(PSQ)评分均高于对照组(均P0.05)。研究组重度职业紧张程度人数占比高于对照组(P0.05)。经多因素Logistic回归分析发现:年龄45岁、受教育程度本科及以上、工龄20年、重度职业紧张均是油田作业人员心理障碍的独立危险因素(均P0.05)。结论:油田作业人员职业紧张状况不容乐观,重度职业紧张、年龄较小、工龄较短及受教育程度较高的油田工作人员发生心理障碍的风险往往较大。临床工作中可通过降低油田作业人员的职业紧张程度,继而达到降低心理障碍发生几率的目的。     

自噬通路基因多态性与晚期非小细胞肺癌含铂化疗疗效的相关性分析

对晚期非小细胞肺癌患者采用含铂化疗是肺癌临床治疗中非常重要的方法,然而不同患者对含铂化疗的敏感性却存在着明显的个体差异,这提示发现潜在的分子标志物对预测临床中含铂化疗疗效具有关键作用。本研究旨在探索自噬通路基因多态性与晚期非小细胞肺癌含铂化疗疗效之间的相关性,以期寻找可能影响含铂化疗药物敏感性的分子标记。本研究纳入了1004例接受含铂化疗的晚期非小细胞肺癌患者,分析了自噬通路中13个基因上的99个SNP位点与含铂化疗临床获益、无疾病进展时间及总生存时间之间的相关性。研究发现,位于ULK1基因的位点rs7953348(G>A) (P=0.017, OR:0.67, 95%CI:0.49~0.93)和rs12303764(A>C) (P=0.009, OR:0.63, 95%CI:0.45-0.89)及位于ATG14基因上的位点rs17742719(C>A) (P=0.002, OR:1.83, 95%CI:1.26~2.66)、rs8003279(A>G) (P=0.006, OR:1.65, 95%CI:1.16~2.35)和rs1009647(G>A) (P=0.002, OR:1.70, 95%CI:1.22~2.37)与临床获益存在显著关联,位于DRAM基因上的位点rs7955890(G>A) (P=0.004, HR:0.63; 95%CI:0.46~0.86)和rs17032060(G>A) (P=0.006, HR:0.65, 95%CI:0.48~0.88)及位于ATG3基因上的位点rs13082005(G>A) (P=0.012, HR:1.27,95%CI:1.05~1.53)与含铂化疗的无疾病进展时间显著相关,位于ULK1基因的位点rs7953348(G>A) (P=0.011, HR:0.74, 95%CI:0.58~0.93)和位于ATG10基因上的位点rs1864183(G>A) (P=0.016, HR:0.42, 95%CI:0.21~0.85)对含铂化疗的总生存时间有着显著影响。研究结果提示自噬通路在含铂化疗敏感性中发挥着重要作用,自噬通路基因多态性可能是预测含铂化疗疗效的潜在分子标志物,这可能为临床上肺癌的个体化医疗提供一定的理论基础。     

单胎妊娠早产胎膜早破呼吸窘迫综合征的危险因素分析

目的:探讨单胎妊娠早产胎膜早破发生新生儿呼吸窘迫综合征(respiratory distress syndrome,RDS)的危险因素。方法:选择2017年5月至2019年5月在我院产科分娩的2810例产妇为研究对象,其中97例(3.45%)符合未足月胎膜早破(Preterm premature rupture of membranes,pPROM)标准,包括53例RDS。收集以下信息:PROM潜伏期、出生时胎龄、脐动脉搏动指数(Umbilical artery pulsatility index,UAPI)、大脑中动脉搏动指数(Middle cerebral artery pulsation index,MCAPI)、胎儿窘迫、产前使用类固醇、新生儿实验室参数、性别、体重、Apgar评分、分娩类型、妊娠高血压疾病、妊娠期糖耐量异常或糖尿病等信息,通过Logistic回归分析研究变量对RDS的影响。结果:Logistic回归分析结果显示,以下变量与RDS密切相关:新生儿性别女性(OR=0.517;95%CI:0.312-0.107;P=0.042),产前使用类固醇(OR=0.467;95%CI:0.355-0.698;P0.001),异常UAPI(OR=2.830;95%CI:1.783-6.234;P=0.002),异常MCA PI(OR=2.136;95%CI:1.120-4.017;P=0.032),胎儿窘迫(OR=2.420;95%CI:1.287-4.824;P=0.017),母体HGB(OR=0.689;95%CI:0.511-1.013;P=0.221),新生儿HGB(OR=0.752;95%CI:0.645-0.891;P0.001),新生儿RBC(OR=0.311;95%CI:0.201-0.565;P0.001)。结论:单胎妊娠早产胎膜早破发生RDS危险因素主要是性别、胎儿胎盘循环异常和胎儿窘迫。     

血站实验室生物安全与防护

血站实验室每天要检测大量无偿献血者的血液样本,而血液样本都具有潜在的生物危害,如乙型肝炎病毒、丙型肝炎病毒、梅毒和艾滋病病毒等。最新研究表明,血站检验人员存在职业暴露的危险因素,极易通过采血针刺伤、试管划伤、血液或粘膜污染等途径感染病毒[1]。随着社会的发展,医疗卫生与健康问题已成为社会关注焦点。所谓实验室生物安全是指在从事病原微生物实验活动的实验室中避免病原微生物对工作人员和相关人员的伤害。为保障实验室工作人员及献血者的安全,血站必须建立符合生物安全要求的实验室,对实验室进行生物安全管理的同时还要重视实验室生物安全防护[2]。本文就血站实验室生物安全及防护措施在人员培训、关键物料、环境和制度等方面提出自己的一些思考和建议。     

油田野外作业人员职业紧张与生理健康及心理障碍的关系研究

目的:探讨油田野外作业人员职业紧张与生理健康及心理障碍的关系。方法:采用分层整群抽样抽取1862例油田野外作业人员,对所有油田野外作业人员发放问卷进行调查,有效回收1546份调查问卷,回收率83.03%。采用职业紧张量表修订表统计油田野外作业人员职业紧张发生状况,其中重度紧张者571例、中度紧张者590例、正常者385例,采用症状自评量表-90(SCL-90)对不同程度职业紧张人员进行心理健康状况评估,采用慢性非传染性疾病调查表问卷统计不同程度职业紧张人员的慢性疾病患病率,采用Logistic回归分析油田野外作业人员心理障碍影响因素。结果:重度紧张者、中度紧张者SCL-90各项评分均高于正常者,且重度紧张者SCL-90各项评分高于中度紧张者(P0.05)。油田野外作业人员心理障碍与其年龄、工龄无关(P0.05),男性、有倒班、吸烟、饮酒情况的油田野外作业人员心理障碍发生率高于女性、无倒班、吸烟、饮酒情况的油田野外作业人员(P0.05)。Logistic回归分析显示,男性、有倒班、吸烟、饮酒情况是影响油田野外作业人员发生心理障碍的危险因素。重度紧张者、中度紧张者的慢性疾病患病率均高于正常者(P0.05)。结论:男性、有倒班、吸烟、饮酒情况的油田野外作业人员心理障碍发生率较高,且油田野外作业人员职业紧张程度越高,越容易增加心理障碍以及慢性疾病的发生风险。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2017年中国植物科学若干领域重要研究进展

2017年中国植物科学继续保持高速发展态势, 重大成果频出, 具体表现在中国植物学家在国际顶级学术期刊发表的文章数量平稳上升。中国植物科学领域的研究工作者成果精彩纷呈, 如新型广谱抗病机制的发现、水稻广谱抗病遗传基础及机制和疫霉菌诱发病害成灾机制研究等。2017年中国生命科学领域十大进展评选中, 有两项植物科学领域的研究成果入选。水稻生物学、进化与基因组学和激素生物学等领域学科发展突出。另外, 值得一提的是, 长期从事高等植物与代谢途径调控分子网络研究和水稻品种设计育种的李家洋院士的研究成果“水稻高产优质性状形成的分子机理及品种设计”荣获2017年国家自然科学一等奖。这一具有重大国际影响的开创性贡献标志着中国植物科学在该领域的国际科学前沿居于引领和卓越地位。该文对2017年中国本土科学家在植物科学若干领域取得的重要研究成果进行了系统梳理, 旨在全面追踪和报道当前中国植物科学领域发展的最新前沿动态, 与广大读者共同分享我国科学家所取得的辉煌成就。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义。综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义.综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望.     

Changes in aquatic vegetation in Rhine floodplain streams in Alsace in relation to disturbance

Abstract. Changes are described in aquatic vegetation in oligotrophic, groundwater-fed Rhine floodplain streams in Alsace (eastern France), resulting from disturbance. Disturbance factors include changes in nutrients, either permanent ones - effluent from a waste water treatment plant or trout hatcheries - or periodic ones: flooding. Regular inputs of high levels of phosphate and ammonia modified the macrophyte vegetation in these streams. The floristic composition, which was characteristic of oligotrophic waters upstream of the eutrophicated sector, changed to that of a eutrophic situation as originally found downstream. Periodic disturbance by floods which normally occur once a year, irregularly eutrophicates the small streams, causing the development of a mixture of eutrophic and oligotrophic species. Six macrophyte communities are distinguished, indicating different trophic levels. The aquatic vegetation is adapted to the variations of phosphate and ammonia levels. Hence, aquatic macrophytes can be used as bio-indicators of fluctuations in water nutrient levels in relation to the type of disturbance.     

Morphine-potentiated platelet aggregation in in vitro and platelet plug formation in in vivo experiments

The detailed mechanisms underlying morphine-signaling pathways in platelets remain obscure. Therefore, we systematically examined the influence of morphine on washed human platelets. In this study, washed human platelet suspensions were used for in vitro studies. Furthermore, platelet thrombus formation induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium was used for an in vivo thrombotic study. Morphine concentration dependently (0.6, 1, and 5 microM) potentiated platelet aggregation and the ATP release reaction stimulated by agonists (i.e., collagen and U46619) in washed human platelets. Yohimbine (0.1 microM), a specific alpha(2)-adrenoceptor antagonist, markedly abolished the potentiation of morphine in platelet aggregation stimulated by agonists. Morphine also potentiated phosphoinositide breakdown and intracellular Ca(2+) mobilization in human platelets stimulated by collagen (1 microg/ml). Moreover, morphine (0.6-5 microM) markedly inhibited prostaglandin E(1) (10 microM)-induced cyclic AMP formation in human platelets, while yohimbine (0.1 microM) significantly reversed the inhibition of cyclic AMP by morphine (0.6 and 1 microM) in this study. The thrombin-evoked increase in pH(i) was markedly potentiated in the presence of morphine (1 and 5 microM). Morphine (2 and 5 mg/g) significantly shortened the time require to induce platelet plug formation in mesenteric venules. We concluded that morphine may exert its potentiation in platelet aggregation by binding to alpha(2)-adrenoceptors in human platelets, with a resulting inhibition of adenylate cyclase, thereby reducing intracellular cyclic AMP formation followed by increased activation of phospholipase C and the Na(+)/H(+) exchanger. This leads to increased intracellular Ca(2+) mobilization, and finally potentiation of platelet aggregation and of the ATP release reaction.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.