无动物细胞条件下麻疹病毒与细菌共生培养的研究

麻疹病毒在无动物细胞的１９９培养基内与甲型链球菌共生的条件下,能自主复制,并可无限地传代。用猴及人的“Ｏ”型红血球凝集试验方法进行实验的结果显示,其血凝滴度可超过１：１０００,抗麻疹病毒免疫血清对所培养的病毒血凝抑制滴度为１：６４。用此病毒免疫家兔所产生的抗体,对麻疹病毒的血凝反应可引起抑制作用。因此,可以说明在培养基内经共生培养所繁殖的病毒的确是麻疹病毒,证实麻疹病毒在无动物细胞的甲型链球菌共生     

甲型流感病毒核糖核蛋白的提纯及其部分理化性质的研究

用Triton x-100和DOC裂解、Sephadex G200柱层析和密度梯度超离心方法,首次从流感病毒感染的鸡胚尿囊膜中提纯了甲型流感病毒RNP。经蛋白质和核酸含量测定、补体结台试验、免疫双扩散及SDS—PAGF鉴定,所提纯的RNP与从病毒中提取的RNP相同。并测出粤防77—38毒株RNP的等电点为4.6,沉降系数为56．1 S。     

昆虫病原线虫和共生细菌培养系统中噬菌体的检测

本文利用紫外照射、温度、pH、盐度诱导五株溶源性共生菌株Xenorhabdus和Photorhabdus,同时测定斯氏和异小杆属线虫Steinernema carpocapsae A24,S.carpocapsae AIL,S.feltiae English,S.feltiae SN,Heterorbabditis bacterophora H06在体外培养过程中是否存在感染共生细菌的噬菌体.结果未发现噬菌斑,说明实验菌株在实验诱导条件下以及昆虫病原线虫固体培养系统中不可能诱发噬菌体的危害.     

甲型流感病毒免疫治疗S_（37）腹水瘤的实验研究

用甲型流感病毒７５－３９株鼠肺适应型,免疫治疗Ｓ３７腹水瘤小鼠,存活率达９３．３％。体外流感病毒感染Ｓ３７肿瘤细胞,经不同时间观测,到３天时Ｓ３７细胞经胎盘蓝染色发现细胞１００％死亡。而对照组Ｓ３７细胞死亡率为１０％左右（ｐ＜０．０１）。进一步研究病毒免疫治疗Ｓ３７腹水瘤小鼠的机理,发现经病毒感染后小鼠ＮＫ细胞杀伤活性升高达５８％,正常鼠ＮＫ活性为２２％,两者有显著性差异（ｐ＜０．０１）。另外,检测病毒注射后小鼠腹腔巨噬细胞的吞噬活性也随之升高。     

石蕊属某些地衣共生菌的培养研究

本文对石蕊属15个种子囊孢子进行了培养特性的研究.所有种子囊孢子均在12小时内萌发,并且前7-15天生长迅速,以后逐渐缓慢.随着培养时间增长.分离物可形成有特征性的菌落形态和色素,利用生长型和菌落及培养基中色素产生情况可将不同分离物区分开.对石蕊属的常规培养很有希望成为识别该属成员的有用特征.     

动物细胞无血清培养基的研究与设计方法

随着哺乳动物细胞培养规模的扩大和生物药物需求的增长,基于细胞及产品特性的无血清培养基的研制已成为细胞工程领域的重要课题。运用统计学实验方法,可科学有效地考察细胞培养基中多因素、多水平间的交互作用。应用新型蛋白质组分析技术及生物芯片技术定位胞浆信号通路相关蛋白、膜表面的生长因子受体、激素受体、细胞因子受体、粘附分子等,用于确定细胞培养基中调控分子的添加组合。本文系统概述了目前无血清培养基几种新型及常用的研究和设计方法,并对其应用特点做了分析,希望为动物细胞无血清培养基的研制提供可借鉴的思路。     

新分离乙型流感病毒的培养温度

乙型流行性感冒病毒在鸡胚培养的温度,各国学者曾作过研究,认为35℃结果最好。我们实验室的分离及传代的常规方法也是培养于35℃。但在1961及1964午乙型流感病毒分离过程中发现,阳性标本的鸡胚羊水或尿液的血凝很不规律,有时甚至全部阴性,特别是冷冻保存(-40℃)的病毒材料,有时盲传二代仍然未见阳性血凝现象。     

链球菌-韦荣菌共生对系统性疾病的诊断价值研究

口腔是人体生理功能的窗口,也是种类和数量繁多的微生物库.口腔微生态变化能够反映宿主与环境因素的相互作用,进而影响机体健康和疾病的进展.其中,链球菌属(Streptococcus)和韦荣菌属(Veillonella)是口腔最早的定殖菌和典型共生菌,共同参与口腔早期生物膜形成.大量研究显示,链球菌和韦荣菌共生失调不仅与龋病...     

喇叭石蕊共生菌藻液体培养的研究

对喇叭石蕊共生菌、藻液体培养条件进行了研究。结果表明:共生菌生长在以40g/L肌醇为碳源、2g/LL-谷氨酰胺为氮源、起始pH值为7.0的LB液体培养基中,培养温度为20℃时表现最佳。其共生藻的生长在以160g/L葡萄糖为碳源、1.75g/LNaNO3为氮源、起始pH值为5.0的BBM液体培养基中,培养温度为20℃时表现最佳。     

住院患者感染甲型流感病毒早期病毒载量与疾病严重程度关系的研究

目的分析住院患者感染甲型流感病毒早期病毒载量与疾病严重程度的关系。方法回顾性分析2014年1月至2015年12月浙江大学医学院附属第一医院收治的甲型流感病毒确诊病例,收集首次痰液标本病毒检测的CT值(与病毒载量呈负相关),以及患者的人口统计学、临床症状和体征等指标信息,以CT值的中位数为界限将患者分成两组进行比较。结果共172例甲型流感病毒(排除确诊的H7N9亚型)感染患者,最常见的临床症状为发热(89.0%)、咳嗽(87.8%)和咳痰(75.6%)。有86.6%的患者肺部影像学检查异常,34.9%的患者接受辅助通气,12.2%的患者被收入ICU,住院时间M=12d(P25~P75:7~22d),9.4%的患者最终死亡。首次甲型流感病毒核酸检测的CT值M=30.1(P25~P75:25.1~34.0),高病毒载量组患者接受辅助通气的比例略高(41.2%vs 28.7%),其中接受侵入性辅助通气的比例显著高于低病毒载量组(20.0%vs 9.2%,P=0.044)。两组患者被收入ICU的比例和住院时间无显著差异,但两组间死亡率(21.2%vs 9.2%)的差异有统计学意义(P=0.034)。结论早期高病毒载量的甲型流感患者病情和预后相对较重较差。     

深圳市龙岗区流感监测及健康人群流感抗体水平分析

对深圳市龙岗区近几年流感病原学及人群流感抗体水平的分析,为流感防控提供科学依据。通过流感监测系统对流感进行病原学和血清学监测的结果显示,2007年流感病毒分离率为1.08%(3/277),2009年分离率为6.68%(56/838),两年流感病毒分离率差别具有统计学意义(χ2=13.03,P=0.000)。2007年健康人群血清样本中H1N1、H3N2、BY、BV流感抗体阳性率分别为60.5%、82.1%、59.3%、50.2%,2010年健康人群血清样本中H1N1、H3N2、BY、BV、甲型H1N1流感抗体阳性率分别为36.2%、37.6%、40.2%、21.9%、37.6%,2007年和2009年各亚型流感抗体阳性率具有统计学意义(P<0.05),两年中均表现H3N2亚型抗体阳性率最高,BV抗体阳性率最低。2007年和2010年H1N1、H3N2、BY、BV抗体阳性率均高于2010年,差别均具有统计学意义(P<0.01)。因此应加强流感病毒抗原变异株和人群流感水平的监测,对5岁以下及20岁以上的易感人群加强流感疫苗的接种,预防流感大流行。     

Binding of bacteria to HEp-2 cells infected with influenza A virus

Epidemiological studies indicate influenza virus infection increases susceptibility to bacterial respiratory pathogens and to meningococcal disease. Because density of colonisation is an important factor in the development of bacterial disease, the objectives of the study were to use flow cytometry methods for assessment of bacterial binding and detection of cell surface antigens to determine: (1) if HEp-2 cells infected with human influenza A virus bind greater numbers of bacteria than uninfected cells; (2) if influenza infection alters expression of cell surface antigens which act as receptors for bacterial binding; (3) if neuraminidase affects binding of bacteria to HEp-2 cells. There was significantly increased binding of all isolates tested regardless of surface antigen characteristics. There were no significant differences between virus-infected and -uninfected Hep-2 cells in binding of monoclonal antibodies to Lewisb, Lewisx or H type 2. There were significant increases in binding of monoclonal antibodies to CD14 (P < 0.05) and CD18 (P < 0.01). Treatment of cells with monoclonal antibodies significantly reduced binding of Neisseria meningitidis strain C:2b:P1.2, CD14 (P < 0.001) and CD18 (P < 0.001). No reduction in binding of a strain of Streptococcus pneumoniae (12F) was observed in these experiments. Neuraminidase treatment of HEp-2 cells increased binding of monoclonal antibodies to CD14 (P < 0.01) and CD18 (P < 0.01). In three experiments, the increase in binding of meningococcal strain C:2b:P1.2 to neuraminidase-treated cells was not significant, but binding of Staphylococcus aureus strain NCTC 10655 was significant (P < 0.05).     

Protection of inactivated influenza virus vaccine against lethal influenza virus infection in diabetic mice

Influenza virus infection frequently causes complications and some excess mortality in the patients with diabetes. Vaccination is an effective measure to prevent influenza virus infection. In this paper, antibody response and protection against influenza virus infection induced by vaccination were studied in mouse model of diabetes. Healthy and diabetic BALB/c mice were immunized once or twice with inactivated influenza virus vaccine at various dosages. Four weeks after the first immunization or 1 week after the second immunization, the mice were challenged with influenza virus at a lethal dose. The result showed that the antibody responses in diabetic mice were inhibited. Immunization once with high dose or twice with low dose of vaccine provided full protection against lethal influenza virus challenge in diabetic mice, however, in healthy mice, immunization only once with low dose provided a full protection.     

流感病毒血凝素蛋白裂解机制的研究进展

血凝素(Hemagglutinin,HA)是流感病毒的主要表面抗原之一,诱导机体产生中和抗体,介导病毒囊膜与靶细胞膜融合,从而启动病毒对宿主细胞的感染过程。HA蛋白以前体形式合成,需经宿主蛋白酶水解为HA1、HA2两个亚单位,并以二硫键连接,病毒才获得感染性。研究表明宿主蛋白酶的分布与流感病毒感染后的致病力和组织嗜性有直接关系。潜在的裂解酶及其抑制因子的发现为流感的防治提供了新的思路,成为干预治疗的新潜在靶点。就当前国内外关于流感病毒血凝素的结构与功能、裂解机制及其应用的研究进展进行综述。     

Diagnosis of influenza viruses with special reference to novel H1N1 2009 influenza virus

On 15 April and 17 April 2009, novel swineorigin influenza A (H1N1) virus was identifi ed in specimens obtained from two epidemiologically unlinked patients in the United States. The ongoing outbreak of novel H1N1 2009 influenza (swine influenza) has caused more than 3,99,232 laboratory confi rmed cases of pandemic influenza H1N1 and over 4735 deaths globally. This novel 2009 influenza virus designated as H1N1 A/swine/California/04/2009 virus is not zoonotic swine flu and is transmitted from person to person and has higher transmissibility then that of seasonal influenza viruses. In India the novel H1N1 virus infection has been reported from all over the country. A total of 68,919 samples from clinically suspected persons have been tested for influenza A H1N1 across the country and 13,330 (18.9%) of them have been found positive with 427 deaths. At the All India Institute of Medical Sciences, New Delhi India, we tested 1096 clinical samples for the presence of novel H1N1 influenza virus and seasonal influenza viruses. Of these 1096 samples, 194 samples (17.7%) were positive for novel H1N1 influenza virus and 197 samples (18%) were positive for seasonal influenza viruses. During outbreaks of emerging infectious diseases accurate and rapid diagnosis is critical for minimizing further spread through timely implementation of appropriate vaccines and antiviral treatment. Since the symptoms of novel H1N1 influenza infection are not specifi c, laboratory confi rmation of suspected cases is of prime importance.     

流行性感冒裂解疫苗的研制

作为换代产品,流行性感冒裂解疫苗的研制已取得突破性进展,根据WHO有关规程的规定和大量的试验研究结果。完整建立了该疫苗的生产工艺和生产质量控制系统,完成了“流行性感冒裂解疫苗制造及检定规程”等规定性文件的起草和审核工作,以中试规模连续生产了三批疫苗并全部自检合格,通过疫苗稳定性试验,效力试验,异常毒性试验及过敏性试验等观察。进一步肯定了疫苗的质量。     

流感病毒对福尔马林敏感性的研究

采用三种流感病毒株：A1/京防/1/86、A3/济防/15/90、A3/武汉/359/95,进行敏感性试验。用不浓度的福尔马林灭活病毒,并在不同间隔时间检测血凝交价及灭活效果。结果显示,三种毒株经福尔马林灭活后,血凝滴度均有下降,病毒灭活时间也随着福尔马林浓度的降低而延长,说明相同亚型不同株别及不同亚型的流感病毒,对福尔马林的敏感程度均有差异。该试验对今后流感病毒的研究及其疫苗生产均有重要意义。     

Influence of the spectrin network on fusion of influenza virus with red blood cells

We examined the influence of the physical state of the membrane skeleton on low pH fusion of influenza virus A/PR 8/34 with intact human red blood cells. Spectrin, the major component of the skeleton, is known to become denaturated at 50°C. After heat treatment of erythrocytes at 50°C we observed an enhanced kinetics of fusion monitored spectrofluorometrically by the octadecylrhodamine fluorescence dequenching assay, while the extent of fusion was not affected. The accelerated fusion of influenza virus after preincubation of red blood cells at 50°C is not mediated by alterations of the lipid phase of the target. From ESR measurements using spin-labelled phospholipids we conclude that heat-induced alterations of the spectrin network did not affect either the phospholipid asymmetry or the fluidity of the exoplasmic and the cytoplasmic leaflets of the erythrocyte membrane. Moreover, as deduced from our previous investigations, the swelling behaviour of red blood cells could not be responsible for the observed effect. Possible mechanisms for the spectrin effect include a change in the ability of the target membrane to bend locally, and a change in the rate of formation and development of the fusion pore.     

细胞适应性流感病毒高产株的制备研究

与鸡胚培养制备的流感疫苗相比,细胞制备的疫苗具有免疫原性好、生产不受鸡胚限制等优点。但目前流感病毒株在细胞上产量较低,成为疫苗生产的主要限制因素。现就用于制备细胞适应性高产株主要的3种方法,即连续传代、随机突变构建病毒突变体和病毒重配的研究进展,以及突变位点对病毒增殖的影响作一概述。