抗新型冠状病毒卵黄抗体对病毒及变异株的体外中和作用

目的 研究抗新型冠状病毒（severe acute respiratory syndrome coronavirus 2，SARS-CoV-2）卵黄抗体（egg yolk immunoglobulin，IgY）在体外对SARS-CoV-2及其变异株的中和抑制作用，探讨其作为鼻腔/口腔喷雾剂用于预防和阻断SARS-CoV-2感染的可能性。方法 采用间接ELISA检测抗SARS-CoV-2 IgY原液及成品对刺突（spike，S）蛋白的抗体效价；用微量细胞病变法检测其对SARS-CoV-2的中和活性；用萤光素酶发光法检测其对SARS-CoV-2假病毒的中和作用。结果 抗SARS-CoV-2 IgY原液的抗S抗原效价为1:32 000~1:64 000，成品的效价为1:16 000~1:32 000；两批原液对活病毒的中和效价分别为1:128和1:256，相应的抑制中浓度（median inhibitory concentration，IC₅₀）分别为36.22和36.50 μg/ml；成品对SARS-CoV-2假病毒的IC₅₀均值分别为4.571 μg/ml（WT）和6.07 μg/ml（D614G）；对变异株假病毒的IC₅₀分别为15.09~29.94 μg/ml（B.1.1.7）、41.71~55.56 μg/ml（B.1.351）和16.66~32.33 μg/ml（B.1.617.2）。结论 抗SARS-CoV-2 IgY与SARS-CoV-2重组S蛋白具有良好的结合活性，在体外能显著地中和SARS-CoV-2活病毒、假病毒及变异株假病毒，有望用于SARS-CoV-2感染的预防和阻断。     

中和抗体检测应用于新型冠状病毒mRNA疫苗效力分析

目的 通过对新型冠状病毒（2019 novel coronavirus,2019-nCoV）mRNA疫苗（简称新冠mRNA疫苗）免疫小鼠后产生的中和抗体进行检测，探索中和抗体检测法应用于mRNA疫苗体内效力评价的可行性。方法 采用6～8周BALB/c小鼠进行后肢肌肉免疫，检测不同免疫剂量和不同免疫程序的中和抗体滴度。并对8家企业生产的新冠mRNA疫苗免疫的小鼠血清进行中和抗体和IgG抗体检测。结果 2 µg、5 µg、10 µg不同剂量mRNA疫苗按照不同免疫程序免疫小鼠后中和抗体检测结果显示，抗体阳转率均为100%，中和抗体反应有明显的剂量-效应关系。2针间隔14 d加强免疫组抗体滴度显著高于间隔7d加强免疫组及一针组（F=57.13, P<0.001）。2 µg剂量间隔7 d加强免疫和间隔14 d加强免疫产生的中和抗体几何平均滴度（geometric mean titer,GMT）分别为218和468，差异有统计学意义（t=3.40， P=0.003）；5 µg剂量间隔7 d加强免疫和间隔14 d加强免疫产生的中和抗体GMT分别为499和1 436，差异有统计学意义（t=3.62，P=0.002）；10 µg剂量间隔7 d加强免疫和间隔14 d加强免疫产生的中和抗体GMT分别为608和1 909，差异有统计学意义（t=3.23，P=0.005）。国内8家企业生产的新冠mRNA疫苗免疫小鼠后均可产生高滴度的IgG抗体(104.5～107.5)和中和抗体(102.6～104.8)，效力测定结果均符合企业质量标准。各企业生产的mRNA疫苗的中和抗体滴度结果差异有统计学意义（F=70.03，P＜0.001）。结论 小鼠免疫后中和抗体检测可用于mRNA疫苗的体内效力评价。     

基于数据库挖掘以血管紧张素转化酶2为靶点治疗新型冠状病毒肺炎的潜在中药成分

目的 利用中药数据库寻找抑制血管紧张素转化酶2（ACE2）治疗新型冠状病毒肺炎（COVID-19）的潜在中药成分。方法 借助SYMMAP、TCMID、ETCM数据库,以ACE2为搜索词,筛选中药成分及中药材,利用GCBI平台来筛选与靶点相关的炎症因子,得到相关疾病及核心网络图。将ACE2输入到STITCH数据库中,查询阳性对照药物。运用Sybyl（2.0）软件处理受体,中药成分与阳性对照药均和ACE2（1R42）及新型冠状病毒（SARS-CoV-2）3CL pro（6LU7）进行对接分析。应用Swiss ADME平台为对接结果较好的中药成分和阳性对照药构建雷达图。结果 从数据库中找到7种具有抑制SARS-CoV-2潜在活性的中药成分和相关中药材。网络分析图中连接基因数最多的是CXCR4基因,它涉及到细胞因子-细胞因子受体相互作用、趋化因子信号通路可能是发挥治疗COVID-19的潜在机制。分子对接的结果表明,2'',4'',3,4-四羟基查尔酮、葛根素与SARS-CoV-2相关蛋白结合最好。结论 2'',4'',3,4-四羟基查尔酮等中药成分可能是治疗COVID-19的潜在成分。     

风疹病毒IgG抗体检测试剂盒国家监督抽验工作

目的：按照国家体外诊断试剂抽验工作方案,评价风疹病毒IgG抗体检测试剂盒（酶联免疫法和化学发光法）的质量。方法：分别使用风疹病毒IgG抗体国家参考品和企业参考品对抽验的试剂盒进行准确性、特异性、检测限和重复性项目检测。结果：使用企业参考品按照产品注册标准进行检测,17批次试剂盒的抽验合格为16批。使用国家参考品检测,检验合格为10批,其中6批次试剂盒将国家阴性参考品检测为阳性,1批次试剂盒将国家检测限参考品检测为可疑。结论：使用国家参考品和企业参考品产生了抽验结果差异,建议在今后国家体外诊断试剂监督抽验工作中,继续探讨使用国家参考品作为统一的评判标准,以加强相关产品的监管工作。     

抗H7N9流感病毒中和抗体快速检测方法的建立及应用

目的：对建立的抗H7N9流感病毒中和抗体快速检测方法进行方法学验证及初步应用。方法：分别采用不同代次细胞对高、中、低不同滴度的阳性血清进行多次平行检测,考察细胞代次对检验结果的影响;采用NIBSC提供的参考品对方法学的特异性进行验证;同时应用抗H7N9的阳性血清检测进一步评估方法的准确性和精密性;采用ELISA-MNT法和血凝抑制（hemagglutination inhibition,HI）试验分别接种H7N9灭活流感疫苗免疫的小鼠血清样本20份,分析两种方法检测结果的相关性。结果：采用ELISA-MNT中和法,使用不同代次MDCK细胞（25、30和35代）检测相同的血清样本的中和抗体滴度结果相同;该方法只对羊抗H7N9的血清具有较高保护力,对其他血清基本没有交叉反应;该方法准确性良好;该方法组间、组内精密性的平均变异系数分别为4%和11%。该方法测定H7N9型流感疫苗免疫后的小鼠血清抗体效价,其结果与HI抗体的相关系数为0.61,表明两种方法的检测结果之间呈良好的正相关性。结论：建立的微量病毒中和法能够满足H7N9流感病毒中和抗体效价检测的要求,可用于H7N9新型大流行流感疫苗的免疫评价。     

抗体阳性与抗体阴性的自身免疫性脑炎患者临床特征比较

目的 比较自身免疫性脑炎（AE）待查患者中,抗体阳性（确诊）的AE患者和抗体阴性（可能）的AE患者之间的临床特征及辅助检查差异。方法 回顾性分析2017年3月至2018年12月于中国科学技术大学附属第一医院诊断为AE待查的患者56例,所有患者的脑脊液和血清进行AE相关抗体[抗N-甲基-D-天冬氨酸受体抗体（NMDAR）、抗α-氨基-3-羟基-5-甲基-4-异噁唑丙酸受体抗体（AMPAR）、抗γ-氨基丁酸B受体抗体（GABA_BR）、抗富亮氨酸胶质瘤失活蛋白1抗体（LGI1）和抗接触素相关蛋白2抗体（CASPR2）]检测。根据抗体检测结果分为抗体阳性的AE患者和抗体阴性的AE患者,其中抗体阳性的AE患者31例,抗体阴性的AE患者25例。比较分析两组患者的临床特征和辅助检查结果的差异,以及疾病严重程度和治疗的差异。结果 31例抗体阳性的患者中,抗NMDA脑炎20例（65%）,抗GABA_BR脑炎9例（29%）,抗LGI1脑炎2例（6%）。抗体阴性的AE患者静脉滴注免疫球蛋白（IVIG）的使用率低于抗体阳性的AE组。两组患者的性别、年龄、前驱症状、神经系统症状、头颅磁共振表现、脑电图结果、脑脊液压力、脑脊液细胞数及脑脊液蛋白水平的差异均无统计学意义（P均>0.05）,两者的入住ICU率、最高mRS评分、出院日mRS评分差异无统计学意义（P均>0.05）。结论 AE患者中抗体阴性的并不少见,抗体阴性的AE患者临床表现与辅助检查跟抗体阳性的AE无显著性差异,对该类患者进行临床表现和辅助检查分析是必要的。     

胶体酒石酸铋胶囊联合美沙拉秦治疗溃疡性结肠炎的临床研究

目的 采用分子对接技术筛选新型冠状病毒S蛋白与血管紧张素转化酶2（ACE2）小分子抑制剂。方法 化合物库采用Selleck中国天然产物库（目录号L1400,2 054种天然产物）。新型冠状病毒S蛋白与ACE2蛋白共晶采用周强研究员课题组发表结果（PDB：6M17）。采用Discovery Studio软件进行分子对接。结果 通过虚拟氨基酸突变实验确定了关键氨基酸,并在此基础上确定了结合空腔,进而从天然化合物库中筛选出11个具有潜在抑制新型冠状病毒S蛋白与ACE2作用的化合物：毛地黄皂苷、灰毡毛忍冬皂苷甲、连翘酯苷B、灰毡毛忍冬皂苷乙、川续断皂苷乙、常春藤苷D、桔梗皂苷D、松果菊苷、人参皂苷Rb₂、人参皂苷Rc、异绿原酸C。结论 从天然产物库中筛选出潜在的新型冠状病毒S蛋白-ACE2小分子抑制剂,为抗新型冠状病毒（SARS-CoV-2）药物研究及处方筛选提供参考。     

人博卡病毒荧光定量PCR方法(TaqMan探针法)的建立及其在血液制品污染情况调查中的应用

目的 开发快捷方便的检测人博卡病毒（human bocavirus,HBoV）方法,并调查华南地区血液制品和原料血浆中的污染情况。方法 建立HBoV的TaqMan探针实时荧光定量PCR方法,并对相关血液制品企业提供的单人份血浆和原料血浆样品、人血白蛋白、静脉注射免疫球蛋白（intravenous immunoglobulin,IVIG）、人纤维蛋白原、人凝血因子Ⅷ及人纤维蛋白胶-人纤维蛋白原中HBoV的污染情况进行检测。结果 HBoV-DNA在原料血浆和人凝血因子Ⅷ样本中的检测率分别为5.2%和5.0%,但在其他血液制品中则没有检出。结论 本研究建立的基于TaqMan探针HBoV的实时荧光定量PCR方法可以很好地检测出原料血浆及血液制品中的HBoV,对于血液制品的病毒安全有重要意义。     

新型冠状病毒肺炎患者恢复期肛拭子中SARS-CoV-2核酸检测结果评价

目的 分析新型冠状病毒肺炎（COVID-19）患者在恢复期肛拭子标本中新型冠状病毒（SARS-CoV-2）核 酸的检出率及其临床价值。方法 对我院确诊的COVID-19患者痰液、咽拭子和肛拭子3种来源的标本同期进行 SARS-CoV-2核酸检测。分析3种标本类型的病毒核酸阳性率及肛拭子病毒核酸阳性时痰液标本和咽拭子标本中 病毒核酸检出情况。结果 共收集50例COVID-19患者的156份样本,其中痰液标本41份,检出SARS-CoV-2阳性 8份（19.5%）;咽拭子标本55份,检出SARS-CoV-2阳性6份（10.9%）;肛拭子标本60份,检出SARS-CoV-2阳性11份 （18.3%）。50例COVID-19患者在恢复期出现病毒核酸阳性者痰液、咽拭子和肛拭子样本核酸检测阳性分别有8例 （19.5%）、6例（12.0%）和10例（20.0%）。10例肛拭子核酸检测阳性患者的呼吸道标本核酸检测显示9例为阴性。结 论 COVID-19患者恢复期肛拭子SARS-CoV-2核酸阳性检出率较咽拭子和痰液标本检出率稍高。患者恢复期的 呼吸道标本SARS-CoV-2核酸连续检测阴性时,应考虑添加肛拭子检测作为出院或者隔离标准。     

清开灵注射液联合亚胺培南西司他丁钠治疗老年重症肺炎的临床研究

目的 基于网络药理学的方法探索银翘解毒软胶囊治疗新型冠状病毒肺炎（COVID-19）的作用机制。方法 将银翘解毒软胶囊所含1 418个化合物与48个COVID-19炎症相关靶蛋白进行分子对接,构建药物-靶蛋白网络,阐明银翘解毒软胶囊的关键活性成分和潜在作用靶点。结果 通过网络分析获得银翘解毒软胶囊的活性成分有50个,主要为黄酮类和三萜类化合物;潜在作用靶点37个,主要为MTOR、JAK3、ACE、ACE2、PIK3CA、TNF、AKT2和MAP2K1等炎症靶点。分子对接结果显示连翘酯苷及牡荆素-2″-O-鼠李糖苷与新型冠状病毒（SARS-CoV-2）3CL水解酶具有较好的亲和力,甘草酸与血管紧张素转化酶II（ACE2）具有较好的亲和力。结论 银翘解毒软胶囊通过干扰SARS-CoV-2病毒复制、调节炎症信号通路表达及炎症因子分泌发挥治疗COVID-19作用。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.