禽流感H9亚型血凝素单克隆抗体的研制和初步应用

目的：获得特异针对禽流感H9亚型血凝素单克隆抗体。方法：以H9N2亚型病毒制成疫苗免疫BALB／c小鼠，取脾细胞与SP2／0细胞融合，取细胞上清用HI筛选。结果：共获得9株稳定分泌针对血凝素蛋白抗体的杂交瘤细胞；经广谱性和特异性鉴定，所有单抗株均能与所有检测的82株不同年代不同地方不同禽源H9亚型流感病毒大部分分离株反应，且所有单抗株均不与非AIV-H9病毒反应，而其中一株（8E6）能与所有检测的H9亚型病毒株反应。进行Western blot分析显示，其中8株单抗能与AIV的Mr为75000处反应，表明是针对AIVH9HA的单抗。结论：本试验获得9株广谱性特异性很好的单抗，特别是8E6株；这些单抗株为以后作病毒抗原性位点分析、临床检测和流行病学调查提供了良好试剂。     

Bronchiolar expression of aquaporin-3 (AQP3) in rat lung and its dynamics in pulmonary oedema

Aquaporins (AQPs) are water channel proteins that permit osmotically driven water movement. To determine their dynamics in pulmonary oedema, we examined the expression of mRNA and protein for AQP1, AQP3, AQP4, and AQP5 in the lungs of normal and thiourea-treated rats. In the thiourea group, lung water content increased significantly (vs. controls) with the peak at around 4 h. Semi-quantitative RT-PCR showed that AQP3 mRNA in the thiourea group rose significantly, peaking at around 4–8 h. The expression of AQP1, AQP4, AQP5, ENaC and CFTR mRNA each decreased significantly some time after the peak in lung water content. Immunoblot analysis showed that glycosylated AQP3 protein was increased 4–10 h after treatment. Expression of the other AQP proteins was not significantly altered, except for that of AQP4. Immunohistochemical examination revealed that AQP1 was expressed in endothelia, AQP3 in the basal cells of the large airways and in cuboidal cells in the bronchioles, AQP4 in the basolateral membrane of airway cells and AQP5 in type-I pneumocytes. Our results suggest that AQP3 is expressed not only in large airways, but also in bronchioles, and is related to water movement in pulmonary oedema.     

人肝癌组织中p53与HSP70相互作用的初步研究

目的 :探讨人肝癌中热休克蛋白 70 (HSP70 )与 p5 3的相互作用。方法 :用免疫组织化学染色法 ,从 12例肝癌组织中筛选HSP70与 p5 3均呈阳性表达的标本 ,并以免疫共沉淀法提取之。然后用SDS PAGE及Westernblot分析双阳性标本中两种蛋白的存在形式。结果 :用免疫组织化学法检测到 12例肝癌组织中有 3例为双阳性 ,用抗HSP70mAb免疫共沉淀的样品 ,可检测到 p5 3蛋白。用抗p5 3mAb免疫共沉淀的样品也可检测到HSP70蛋白。结论 :人肝癌中p5 3与HSP70以复合物的形式而存在 ,此可为肝癌的发病机制及免疫治疗的研究提供新的思路     

Three cases of childhood nocturnal asthma due to buckwheat allergy

BACKGROUND: Buckwheat flour (BF) is known as a potent food allergen. Sensitization to it usually occurs by ingestion but also by inhalation in occupational or domestic exposure. Buckwheat chaff-stuffed pillows (BCP) can be contaminated with BF during the process of pilling, and many Korean children and adults use BCP for health reasons. METHODS AND RESULTS: We here present three cases of BF allergy in children using BCP, who had been treated as nonatopic asthmatics after undergoing the routine allergy skin tests and serologic tests. We took careful clinical histories, and performed skin prick tests (SPT), the radioimmunoassay (RIA) for specific IgE, the BCP-elimination test, the BF bronchial provocation test, and IgE Western blot. All three children showed positive skin reactions to BF, but none of them had positive reactions to house-dust mites. Nocturnal asthmatic symptoms were improved during 7 days of BCP elimination, and asthmatic responses were observed by bronchial provocation tests with homemade BF extract. Serum BF-specific IgE antibodies and several IgE-binding proteins were detected by RIA and Western blot analysis, respectively. CONCLUSIONS: Thus, a small amount of BF attached to BCP can induce BF sensitization, and BCP should be considered a main cause of childhood nocturnal asthma in those asthmatics exposed to these pillows.     

人血管抑素A K(1-3)基因的克隆表达、纯化及活性鉴定

目的 克隆人Angiostatin K(1-3)基因,获得有活性的重组人血管抑素蛋白,为进一步开发应用奠定基础.方法 以人新鲜肝脏组织为材料,通过RT-PCR得到人Angiustatin基因的AK(1-3)片段.构建重组质粒pET30a-Angiostatin,转化表达菌Rosetta(DE3),对转化子进行诱导表达,利用Ni-NTA亲和层析纯化目的 产物,并验证其活性.结果 获得了人Angiostatin基因AK(1-3)片段的正确序列,表达和纯化了人血管抑素蛋白,表达量占菌体总蛋白的30%,纯化后证明表达产物具有较高纯度(达到90%).结论 Anginstatin K(1-3)可在原核融合蛋白表达载体中表达,且得到有活性的目的 蛋白.     

Blocking-ELISA for detection of specific antibodies to the glycoproteins of the human immunodeficiency virus type 1 (HIV-1)

A blocking ELISA was developed to confirm the specificity of screening tests for anti-HIV-1 antibodies. A murine monoclonal antibody (McAb) raised against recombinant gp160 was used in combination with a commercial technique (ELA-VIA-1). After determining the optimal experimental conditions, the assay was applied to 92 samples presenting different reactivities by Western blot (WB) analysis. All the sera containing antibodies to gp160/gp120 (53) were positive in our assay. The six patients who sero converted showed a low positivity by ELAVIA-1 (optical density near the cutoff value) reacted by blocking-ELAVIA-1 with an McAb binding inhibition greater than 85%. By contrast, negative samples (29) and specimens that exhibited reactivity only against gag-proteins (10) were not detected (McAb binding inhibition smaller than 15%). This sensitive and specific blocking-ELAVIA-1 represents a convenient alternative to WB as a confirmatory test. The technique is time-saving and inexpensive and can easily be integrated with a screening test for diagnostic or epidemiologic studies on HIV-1 infection.     

A Cross-Sectional,Correlational Study Comparing Individual Characteristics of Younger and Older Nursing Home Residents using Western Canadian Resident Assessment Instrument–Minimum Data Set (RAI-MDS) 2.0

ObjectiveTo compare characteristics of nursing home (NH) residents by age categories in Western Canada.DesignA cross-sectional, correlational analysis of secondary data.Setting and Participants89,231 residents living in Western Canada NHs in the provinces of Alberta, Manitoba, and British Columbia in 2016 and 2017.MethodsResident characteristics (age, sex, marital status, body mass index, medical diagnoses, cognitive function, physical function, depressive symptoms) came from the Resident Assessment Instrument–Minimum Data Set 2.0 and were analyzed using chi-square, analysis of variance, and post hoc pairwise tests. Human developmental stage age categories were used to create 5 age groups: 18-34, 35-50, 51-64, 65-80, and 81 years and older.ResultsThe demographics, medical diagnoses, cognitive function, and physical function characteristics of NH residents among 5 age groups differed considerably (all P < .001). Residents aged 18-34 years were predominately male, never married, with a higher incidence of paralysis and traumatic brain injury. Residents aged 35-50 years had a higher incidence of stroke and multiple sclerosis, and residents aged 51-64 years mainly were morbidly obese and more prone to depression. Residents aged 65-80 years were predominately married and more prone to diabetes, and residents aged 81 years and older were predominately widowed, with a higher incidence of dementia compared with others.Conclusions and ImplicationsFindings describe the uniqueness of younger NH age groups and indicate that the youngest NH residents often have the severe disability and a modest support system (as defined by partnered status) compared to older residents in NHs. Future studies must analyze longitudinal data that track the growth of, and changes in, residents’ health and functional status.     

中西医结合治疗眼底病的实验和临床研究

眼底病是一种复杂的致盲率很高的眼部疾病 ,包括视网膜脱离、视网膜玻璃体出血和增殖性玻璃体视网膜病变 (PVR)等疾病。本文采用中西医结合的治疗方法对动物模型和 1998～ 2 0 0 0年此类眼病患者进行研究 ,疗效显著 ,使眼底病的治愈率有了很大幅度的提高。中医药对眼底病的治疗常用活血化瘀复方治疗 ,对于从动物实验及临床两方面同时进行中西医结合的系统治疗研究 ,尚未见有报道。     

中西医结合治疗老年慢性肾功能衰竭

目的观察中西医结合治疗老年慢性肾功能衰竭的临床疗效.方法老年慢性肾功能衰竭病人57例,随机分为治疗组和对照组.对照组用常规西药治疗,治疗组在上述治疗基础上加用降压排毒饮和中药灌肠,观察治疗前后血压、尿素氮、血肌酐、内生肌酐清除率及血脂变化.结果治疗组经治疗后血压、尿素氮、血肌酐、胆固醇、甘油三酯显著下降,而内生肌酐清除率、高密度脂蛋白上升;对照组除血压略下降,其他无显著变化,两组比较,有显著差异,P＜0.01.结论中西医结合疗法在降压、降脂、改善肾功能方面,均显著优于对照组.     

缺氧预适应小鼠脑组织中缺氧诱导因子-1的表达

为探讨缺氧诱导因子 1 (HIF 1 )在缺氧预适应小鼠脑组织中的表达 ,用Western印迹法检测慢性缺氧 (H)和不缺氧 (C)HeLa细胞、急性重复性缺氧 0次 (H0 )、1次 (H1)、4次 (H4 )小鼠脑组织中的HIF 1α。结果 :慢性缺氧HeLa细胞中可检测到较多的HIF 1α,H0 组脑组织中检测不到HIF 1α ,H1组检测到微量HIF 1α,H4 组检测到较多HIF 1α。结果显示 :脑组织中HIF 1α的含量随缺氧预适应的产生而逐步增加 ,提示HIF 1可能参与缺氧预适应的形成。