牛海绵状脑病的自身免疫学说

本文从三个方面叙述牛海绵状脑病（ＢＳＥ）的自身免疫学说;（１）Ｐｒｉｏｎ理论的诘难;（２）ＢＳＥ发病的自身免疫学说的提出;（３）ＢＳＥ是自身免疫性疾病学说的证据和可能机制。最后,对ＢＳＥ是自身免疫性学说作了评价。     

海绵状血管瘤的MRI诊断

本文总结了我院自89年以来的24例经手术和病理证实的海绵状血管瘤,就期MRI表现特征,诊断与鉴别诊断要点作了较详细的分析研究。认为MRI对此病的检出率及定位正确性有着传统方法及CT不可比拟的优点,但其定性诊断的正确率有待进一步提高。随着MRI仪的不断普及和放射科医生经验的不断积累,人们对海绵状血管瘤这一少见的血管性病变的认识一定会不断提高。     

免疫印迹法检测牛海绵状脑病和羊瘙痒病

用大肠杆菌表达的牛朊病毒正常成熟蛋白 (BoPrPC)免疫新西兰白兔 ,获得了与朊病毒蛋白 (PrP)反应的抗体T1。根据致病型朊病毒 (PrP^SC)能抵抗蛋白酶消化的特性 ,用蛋白酶K消化脑组织提取物 ,以抗体T1进行免疫印迹反应 ,结果表明从接种羊瘙痒病朊病毒 2 6 3K的金黄地鼠脑组织提取物内检测到抗蛋白酶K消化的致病型PrPSC ,而正常金黄地鼠脑组织中没有抗蛋白酶消化的蛋白。以我国正常牛羊为材料 ,制备其脑组织提取物 ,用上述方法和抗体T1进行检测 ,结果没有发现抗蛋白酶K的任何蛋白存在 ,说明没有牛海绵状脑病和羊瘙痒病存在。用 1A8抗体也获得了同样的结果。这些结果表明可以用自制的抗血清检疫牛海绵状脑病和羊瘙痒病 ,防止其传入我国     

传染性海绵样脑病病原的研究进展

传染性海绵样脑病因疯牛病的爆发和新型克雅氏病的出现而引起人们的关注。研究发现,疯牛病的流行可能是由羊痒病的病原引起的,这病原被Ｐｒｕｓｉｎｅｒ命名为“Ｐｒｉｏｎ”（１９８２）。从８０年代开始研究以来,越来越多的关于Ｐｒｉｏｎ的特性被人们所了解,比如Ｐｒｉｏｎ蛋白和ＰｒＰ基因。本文综述了近几年来研究者们用不同手段从各个角度对病原进行研究的新进展。     

患羊痒疫及牛海绵状脑病感染羊脑中PrPd的不同免疫反应性

本实验应用多种抗PrP^d的单克隆抗体来研究分别感染羊痒疫或牛海绵状脑病(BSE)病原的羊脑中PrP^d蛋白沉积类型的差异。     

胰性脑病的诊断治疗

胰性脑病是急性胰腺炎的罕见并发症,诊断治疗困难,死亡率极高。本文详细讨论该病的发病机制、临床表现、诊断、鉴别诊断、预防及治疗方面的进展。     

深部念珠菌感染的诊断进展

近年来,真菌感染的发病率和病死率不断上升。念珠菌作为常见的机会性致病真菌,其感染的早期诊断目前存在很多难点。随着人们对念珠菌认识的不断深入和诊断方法的不断进步,深部念珠菌感染的早期诊断在过去几年中取得了很大的发展。现就深部念珠菌感染的诊断进展做一综述。     

克雅氏病———医患双方的挑战

克雅氏病是由朊蛋白病毒感染人类引起的罕见的、可传播性的、致命性的神经系统疾病,目前已成为令全球恐慌的不可治愈性疾病,其患者在全球范围的增加,给医患双方带来新的挑战.本文就克雅氏病的临床分型、流行病学、病理、诊断及护理作一系统介绍,使大家对该病的认识更加深刻.     

脑海绵状血管瘤的MRI与临床特征及其治疗（附二例报告）

目的：探讨脑海绵状血管瘤的临床特征。方法：回顾分析29例脑海绵状血管瘤的临床资料,对其临床表现、诊断和治疗进行总结。结果：主要临床表现为头痛和癫痫发作,删有助于明确诊断。结论：MRI是诊断脑海绵状血管瘤最敏感的方法,对有症状施行积极的显微外科治疗能收到良好效果。     

Prion diseases: pathogenesis and public health concerns

Transmissible spongiform encephalopathy (TSE) agents or prions induce neurodegenerative fatal diseases in humans and in some mammalian species. Human TSEs include Creutzfeldt-Jakob disease (CJD), Gerstmann-Str?ussler-Scheinker syndrome, kuru and fatal familial insomnia. In animals, scrapie in sheep and goats, feline spongiform encephalopathy, transmissible mink encephalopathy, chronic wasting disease in wild ruminants, and bovine spongiform encephalopathy (BSE), which appeared in the UK in the mid-1980s [Wells, G.A.H. et al. (1987) Vet. Rec. 121, 419-420], belong to the TSE group. Prions have biological and physicochemical characteristics that differ significantly from those of other microorganisms; for example, they are resistant to inactivation processes that are effective against conventional viruses, including those that alter nucleic acid structure or function. Alternatively, infectivity is highly susceptible to procedures that modify protein conformation. Today, the exact nature of prions remains unknown even though it is likely that they consist of protein only. At the biochemical level, TSEs are characterised by the accumulation, within the central nervous system of the infected individual, of an abnormal isoform of a particular protein from the host, the prion protein [Prusiner, S.B. (1982) Science 216, 136-144]. TSEs are transmissible among their species of origin, but they can also cross the species barrier and induce chronic infection and/or disease in other species. Transmissibility has been proven in natural situations such as the outbreak of CJD among patients treated with pituitary-derived hormones and the appearance of BSE that affected UK cattle in the mid-1980s.     

Concentration of Disease-Associated Prion Protein with Silicon Dioxide

Reagents that can precipitate the disease-associated prion protein (PrP^Sc) are vital for the development of high sensitivity tests to detect low levels of this disease marker in biological material. Here, a range of minerals are shown to precipitate both ovine cellular prion protein (PrP^C) and ovine scrapie PrP^Sc. The precipitation of prion protein with silicon dioxide is unaffected by PrP^Sc strain or host species and the method can be used to precipitate bovine BSE. This method can reliably concentrate protease-resistant ovine PrP^Sc (PrP^res) derived from 1.69 μg of brain protein from a clinically infected animal diluted into either 50 ml of buffer or 15 ml of plasma. The introduction of a SiO₂ precipitation step into the immunological detection of PrP^res increased detection sensitivity by over 1,500-fold. Minerals such as SiO₂ are readily available, low cost reagents with generic application to the concentration of diseases-associated prion proteins.     

Transmissible spongiform encephalopathy strain-associated diversity of N-terminal proteinase K cleavage sites of PrPSc from scrapie-infected and bovine spongiform encephalopathy-infected mice

Assessment of the different conformational states of the abnormal prion protein (PrP^Sc) in the CNS provides an established basis for distinguishing transmissible spongiform encephalopathy (TSE) strains. PrP^Sc conformers are variably resistant to N-terminal proteinase K (PK) digestion, and analysis of the consensus products (PrP^res) by immunoassay enables effective, but relatively low-resolution differentiation. Determination of the precise N-terminal amino acid profile (N-TAAP) of PrP^res presents a potential high-resolution means of TSE-strain typing, and thus of differential disease diagnosis. This approach was evaluated using individual mice affected by model scrapie (22A, ME7, 87V and 79A) and bovine spongiform encephalopathy (BSE) (301V) strains. Nano liquid chromatography–mass spectrometry (LC-MS) was used to determine PrP^res N-terminal tryptic digestion products. Four major N-terminal tryptic peptides were generated from all mouse TSE strains investigated, corresponding with predominant N-termination of PrP^res at G⁸¹, G⁸⁵, G⁸⁹ and G⁹¹. Both the mass spectrometric abundance of the individual peptides and the ratios of pairs of these peptides were evaluated as markers of conformation in relation to their potential for strain discrimination. The yield of peptides was significantly greater for BSE than scrapie strains and the relative quantities of particular peptide pairs differed between strains. Thus, whereas peptide G⁹¹–K¹⁰⁵ was a dominant peptide from 301V, this was not the case for other strains and, significantly, the ratio of peptides G⁹¹–K¹⁰⁵:G⁸⁹–K¹⁰⁵ was substantially higher for BSE-infected compared with scrapie-infected mice. These data support the potential of the N-TAAP approach for high-resolution TSE strain typing and differential diagnosis.     

Inhibition of PrPSc formation by synthetic O-sulfated glycopyranosides and their polymers

Sulfated glycosaminoglycans (GAGs) and sulfated glycans inhibit formation of the abnormal isoform of prion protein (PrPSc) in prion-infected cells and prolong the incubation time of scrapie-infected animals. Sulfation of GAGs is not tightly regulated and possible sites of sulfation are randomly modified, which complicates elucidation of the fundamental structures of GAGs that mediate the inhibition of PrPSc formation. To address the structure-activity relationship of GAGs in the inhibition of PrPSc formation, we screened the ability of various regioselectively O-sulfated glycopyranosides to inhibit PrPSc formation in prion-infected cells. Among the glycopyranosides and their polymers examined, monomeric 4-sulfo-N-acetyl-glucosamine (4SGN), and two glycopolymers, poly-4SGN and poly-6-sulfo-N-acetyl-glucosamine (poly-6SGN), inhibited PrPSc formation with 50% effective doses below 20 microg/ml, and their inhibitory effect became more evident with consecutive treatments. Structural comparisons suggested that a combination of an N-acetyl group at C-2 and an O-sulfate group at either O-4 or O-6 on glucopyranoside might be involved in the inhibition of PrPSc formation. Furthermore, polymeric but not monomeric 6SGN inhibited PrPSc formation, suggesting the importance of a polyvalent configuration in its effect. These results indicate that the synthetic sulfated glycosides are useful not only for the analysis of structure-activity relationship of GAGs but also for the development of therapeutics for prion diseases.     

A study of neurological diseases in farmed deer in Switzerland,with emphasis on chronic wasting disease

A study of neurological diseases in farmed deer, with emphasis on chronic wasting disease, was conducted during 2 years in Switzerland. Deer breeders were asked to submit the heads of all deer at least 2 years of age, found dead or slaughtered, for examination. A complete histological examination of the brain and immunohistochemical detection of the prion protein on selected regions of the brain and lymphoid tissues were performed on 120 apparently healthy and 40 diseased animals. In a number of cases, a full necropsy was performed. Significant inflammatory and/or degenerative changes were seen in 25% of the brains. No evidence for a transmissible spongiform encephalopathy was established.     

PrPC interacts with tetraspanin-7 through bovine PrP154-182 containing alpha-helix 1

The cellular prion protein (PrP^C) is highly conserved in the evolution of mammals, and therefore, thought to have important cellular functions. Despite decades of intensive research, the physiological function of PrP^C remains enigmatic. We carried out a yeast two-hybrid screen on a bovine brain cDNA expression library and identified the transmembrane protein tetraspanin-7 (CD231), as a PrP^C interacting protein. We confirmed the interaction between PrP^C and tetraspanin-7 by yeast two-hybrid assay, immunofluorescent co-localization, and immunocoprecipitation. Our mutational studies further demonstrated that PrP^C specifically binds tetraspanin-7 through the region corresponding to bovine PrP_154-182 containing alpha-helix 1.     

Polymorphisms of Two Indels at the PRNP Gene in Three Beef Cattle Herds

Bovine spongiform encephalopathy (BSE) is a transmissible fatal neurodegenerative disorder, presenting a characteristic spongiform degeneration of cattle brain due to the accumulation of a pathogenic and protease-resistant infectious protein (prion). Two deletion/insertion polymorphisms of the prion protein gene (23 bp at the promoter region and 12 bp at intron 1) were analyzed in three beef cattle herds (Aberdeen Angus, Charolais, and Franqueiro) to verify allele frequencies for possible use in selection of resistant animals. High frequencies of susceptibility alleles (23 and 12 bp deletion) and haplotype (23 del/12 del) were observed in the Aberdeen Angus and Charolais herds, but Franqueiro presented one of the highest frequencies of resistant alleles so far described. These data indicate the need for selection in Aberdeen Angus and Charolais breeds to increase the frequency of resistant animals in order to reduce the probabilities of BSE outbreaks in these populations.     

An unusual soluble beta-turn-rich conformation of prion is involved in fibril formation and toxic to neuronal cells

A key molecular event in prion diseases is the conversion of the prion protein (PrP) from its normal cellular form (PrPC) to the disease-specific form (PrPSc). The transition from PrPC to PrPSc involves a major conformational change, resulting in amorphous protein aggregates and fibrillar amyloid deposits with increased beta-sheet structure. Using recombinant PrP refolded into a beta-sheet-rich form (beta-PrP) we have studied the fibrillization of beta-PrP both in solution and in association with raft membranes. In low ionic strength thick dense fibrils form large networks, which coexist with amorphous aggregates. High ionic strength results in less compact fibrils, that assemble in large sheets packed with globular PrP particles, resembling diffuse aggregates found in ex vivo preparations of PrPSc. Here we report on the finding of a beta-turn-rich conformation involved in prion fibrillization that is toxic to neuronal cells in culture. This is the first account of an intermediate in prion fibril formation that is toxic to neuronal cells. We propose that this unusual beta-turn-rich form of PrP may be a precursor of PrPSc and a candidate for the neurotoxic molecule in prion pathogenesis.     

Alkaline serine protease produced by Streptomyces sp. degrades PrP(Sc)

A PrP(Sc)-degrading enzyme was isolated from the culture medium of Streptomyces sp. using perchloric acid-soluble protein (PSP) as a substrate. The media of 500 microbial species were screened to obtain the PSP-degrading enzyme. The medium containing the protease secreted from strain 99-GP-2D-5 showed the highest PSP-degrading activity. Strain 99-GP-2D-5 was assigned as the genus Streptomyces by its morphological and chemotaxonomic characteristics. When scrapie prion was used as the substrate, it was completely digested by the enzyme. The amino acid sequence of the enzyme was identical to that of the C-terminal region of alkaline serine protease (ASP) I. ASP I may be the precursor of the enzyme, and the enzyme seems to be the mature type of ASP I. The maximal activity of the enzyme was observed at 60 degrees C and pH 11, and the scrapie prion was degraded within 3 min under the optimum conditions.     

A role for His155 in binding of human prion peptide144-167 to immobilised prion protein

The interactions and conformational changes that lead to the conversion of the normal prion protein (PrP(c)) to its pathogenic form, PrP(sc), are still being elucidated. Using Surface Plasma Resonance (SPR), we provide evidence that a synthetic peptide (PrP(144-167)) corresponding to residues comprising the alpha helix 1-beta strand 2 domain of PrP(c) is able to interact and bind to immobilised recombinant human PrP (rHuPrP) in a dose-dependent manner. The interaction is pH dependent with an increase in binding observed as the pH is lowered, particularly between pH 6.5 and pH 5.5 suggesting a specific role for His(155) in the interaction, confirmed by covalent modification of this residue in the peptide with diethylpyrocarbonate (DEPC). Circular dichroism analysis of PrP(144-167) revealed no secondary structure motifs across the pH range investigated. Possible pH related structural changes of immobilised rHuPrP are also discussed with regard to the increased affinity for PrP(144-167).