蓝氏贾第鞭毛虫α-19贾第素的单克隆抗体制备与亚细胞定位研究

目的 制备蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)α-19贾第素的特异性抗体,并通过免疫荧光对该贾第素的亚细胞定位进行鉴定。方法 经PCR获得α-19贾第素编码区片段,双酶切将目的 片段连入原核表达载体pET-28a(+)-α-19,重组载体转化E.coli Rosetta(DE3),IPTG诱导表达,SDS-PAGE及 Western blot验证表达情况;采用设计的合成抗原肽免疫小鼠经细胞融合和筛选制备特异性单克隆抗体,分别用贾第虫滋养体裂解物和α-19贾第素重组蛋白Western blot验证抗体特异性和结合力,选择特异性最好的单克隆抗体通过免疫荧光技术对α-19贾第素的亚细胞定位进行观察。结果 成功构建了原核表达载体pET-28a(+)-α-19,经SDS-PAGE及 Westernblot分析显示重组载体在大肠杆菌中表达出相对分子量约49 kD的重组蛋白,与理论值相符;Western blot显示单克隆杂交瘤细胞株α-19-3-7-3-5分泌的抗体结合力和特异性俱佳,可用于定位研究;免疫荧光定位结果表明α-19贾第素主要位于贾第虫滋养体的腹鞭毛。结论 制备了α-19贾第素的特异性单克隆抗体,免疫荧光显示α-19贾第素定位于贾第虫滋养体的腹鞭毛。     

电视胸腔镜手术前CT引导下亚甲蓝和/或Hookwire定位肺磨玻璃结节

目的 探讨肺磨玻璃结节（GGN）电视胸腔镜手术（VATS）前CT引导下亚甲蓝染色和/或Hookwire定位结节的临床价值。方法 对141例肺GGN患者（160枚结节）均在CT引导下亚甲蓝染色和/或Hookwire定位结节后行VATS,计算定位成功率、穿刺并发症发生率及手术成功率。结果 141例均成功定位并切除肺GGN。单独亚甲蓝染色定位18枚,Hookwire定位12枚,二者联合定位130枚,定位成功率均为100%。穿刺过程中少量肺出血25例,气胸38例,肺出血并发气胸13例。术后病理示恶性结节117枚,良性结节43枚。结论 对肺GGN行VATS术前CT引导下亚甲蓝染色和/或Hookwire均可准确定位结节,有助于减少穿刺并发症、提高切除成功率。     

Structural characterisation, stability and antibody recognition of chimeric NHBA-GNA1030: an investigational vaccine component against Neisseria meningitidis

A new generation multi-component vaccine, principally directed against serogroup B Neisseria meningitidis (4CMenB), has recently been developed. One of its components, identified through reverse vaccinology, is the neisserial heparin-binding antigen (NHBA) which is included in the formulation as a novel NHBA-GNA1030 fusion protein (NHBA-FP). We describe here the biophysical characteristics of this vaccine antigen to understand better its structural properties in solution and concurrent immunogenicity prior to formulation. By deliberately stressing the protein to lose its immune responses, we were able to study the protein's structural changes at the molecular level. The unmodified NHBA-FP was found to be mainly monomeric with mass of 67kDa and secondary structure dominated by β-sheets and turns (57% average). The antigen was very stable in storage buffer. It could be forced to unfold in a low-salt buffer resulting in the exposure of one of its two tryptophan residues at 50°C. Long-term stress studies (10-15 days at 37°C) showed modification in the chromatographic and electrophoretic profiles with progressive degradation and aggregation. Since there was little change in secondary structure (as monitored by circular dichroism and tryptophan fluorescence spectroscopy), the loss of functional immunogenicity of the thermal stressed protein could be mainly attributed to the observed fragmentation and aggregation. We therefore conclude that the maintenance of the intact, non-fragmented state of the NHBA-FP is important to preserve its functional immunogenicity. This may thus be utilised as an assay for the control testing of the protein.     

细针穿刺定位活检对诊断早期乳腺癌的意义

目的 探讨细针穿刺导丝定位活检(FNL)对诊断早期乳腺癌的意义。方法 28例临床触诊阴性、乳腺钼靶照相可疑病变的患者，根据轴位及侧位摄片显示的病灶位置，将穿刺细针穿入，并将导丝留置于病变内，随后外科医师沿留置的导丝将病灶切除，行病理检查。结果 5例病灶病理诊断为原位癌，1例浸润癌，阳性率为23．1％，患者行局部放射治疗，一年后未见转移及复发。1例轻度非典型增生伴导管内乳头状瘤，另6例为良性病变，14例为乳腺组织增生，1例穿刺失败。结论 细针穿刺定位活检不仅可以提高早期乳腺癌的诊断率，而且可以协助治疗。     

Does the agglomeration effect occur in the hospital sector? The impact of agglomeration economies on the financial performance of hospitals—An evidence from Poland

The main aim of the article is to analyze the occurrence of agglomeration effect in the hospital sector on the basis of financial performance. The considerations are made on the example of hospitals in Poland—the country that survived the latest economic crisis relatively well, usually generating positive values of GDP, but where still there is an ongoing discussion on the final shape of healthcare financing model. The article is based on the assumption that there occur significant differences in financial performance between hospitals according to their location. The research hypothesis is as follows: Hospitals operating in big cities are featured by better financial condition than their counterparts operating in smaller towns. To verify the hypothesis, the methods of financial analysis and statistical hypothesis testing are used. As it is emphasized in the article, the assumption is true and the hypothesis can be verified positively.     

Early unilateral cochlear implantation promotes mature cortical asymmetries in adolescents who are deaf

Unilateral cochlear implant (CI) stimulation establishes hearing to children who are deaf but compromises bilateral auditory development if a second implant is not provided within ～1.5 years. In this study we asked: 1) What are the cortical consequences of missing this early sensitive period once children reach adolescence? 2) What are the effects of unilateral deprivation on the pathways from the opposite ear? Cortical responses were recorded from 64‐cephalic electrodes within the first week of bilateral CI activation in 34 adolescents who had over 10 years of unilateral right CI experience and in 16 normal hearing peers. Cortical activation underlying the evoked peaks was localized to areas of the brain using beamformer imaging. The first CI evoked activity which was more strongly lateralized to the contralateral left hemisphere than normal, with abnormal recruitment of the left prefrontal cortex (involved in cognition/attention), left temporo‐parietal‐occipital junction (multi‐modal integration), and right precuneus (visual processing) region. CI stimulation in the opposite deprived ear evoked atypical cortical responses with abnormally large and widespread dipole activity across the cortex. Thus, using a unilateral CI to hear beyond the period of cortical maturation causes lasting asymmetries in the auditory system, requires recruitment of additional cortical areas to support hearing, and does little to protect the unstimulated pathways from effects of auditory deprivation. The persistence of this reorganization into maturity could signal a closing of a sensitive period for promoting auditory development on the deprived side. Hum Brain Mapp 37:135–152, 2016. © 2015 Wiley Periodicals, Inc.     

Intracranial EEG potentials estimated from MEG sources: A new approach to correlate MEG and iEEG data in epilepsy

Detection of epileptic spikes in MagnetoEncephaloGraphy (MEG) requires synchronized neuronal activity over a minimum of 4cm2. We previously validated the Maximum Entropy on the Mean (MEM) as a source localization able to recover the spatial extent of the epileptic spike generators. The purpose of this study was to evaluate quantitatively, using intracranial EEG (iEEG), the spatial extent recovered from MEG sources by estimating iEEG potentials generated by these MEG sources. We evaluated five patients with focal epilepsy who had a pre‐operative MEG acquisition and iEEG with MRI‐compatible electrodes. Individual MEG epileptic spikes were localized along the cortical surface segmented from a pre‐operative MRI, which was co‐registered with the MRI obtained with iEEG electrodes in place for identification of iEEG contacts. An iEEG forward model estimated the influence of every dipolar source of the cortical surface on each iEEG contact. This iEEG forward model was applied to MEG sources to estimate iEEG potentials that would have been generated by these sources. MEG‐estimated iEEG potentials were compared with measured iEEG potentials using four source localization methods: two variants of MEM and two standard methods equivalent to minimum norm and LORETA estimates. Our results demonstrated an excellent MEG/iEEG correspondence in the presumed focus for four out of five patients. In one patient, the deep generator identified in iEEG could not be localized in MEG. MEG‐estimated iEEG potentials is a promising method to evaluate which MEG sources could be retrieved and validated with iEEG data, providing accurate results especially when applied to MEM localizations. Hum Brain Mapp 37:1661–1683, 2016. © 2016 Wiley Periodicals, Inc .     

7 tesla T2*‐weighted MRI as a tool to improve detection of focal cortical dysplasia

Focal cortical dysplasia is one of the most common underlying pathologies in patients who undergo surgery for refractory epilepsy. Absence of a MRI‐visible lesion necessitates additional diagnostic tests and is a predictor of poor surgical outcome. We describe a series of six patients with refractory epilepsy due to histopathologically‐confirmed focal cortical dysplasia, for whom pre‐surgical 7 tesla T₂*‐weighted MRI was acquired. In four of six patients, T₂* sequences showed areas of marked superficial hypointensity, co‐localizing with the epileptogenic lesion. 7 tesla T₂* hypointensities overlying focal cortical dysplasia may represent leptomeningeal venous vascular abnormalities associated with the underlying dysplastic cortex. Adding T₂* sequences to the MRI protocol may aid in the detection of focal cortical dysplasias.