重组人脑髓鞘碱性蛋白及其抗体的研究

将EcoR1和SalⅠ酶切的人脑髓鞘碱性蛋白（MBP）基因cDNA克隆片段与表达载体pGEX-5T重组后转化大肠杆菌，经筛选，增殖和IPTG诱导，阳性克隆SDS－PAGE结果证实表达一条特异42KDa区带，Western印记杂交证实该区带具MBP抗原特异性，免疫斑点杂交和ELISA检测表达产量占菌体可溶性蛋白含量6％，达到414.6mg/L菌液，将含可溶性MBP蛋白的菌液后SDS－PAGE分离纯化得重组MBP抗原，对新西兰兔进行背部皮下多点注射，5次免疫后以琼脂板免疫双扩法检测抗效价达1：16，并通过免疫斑点杂交和Western印亦杂交证实证抗体具抗MBP特异性。     

In vivo and in vitro effects of imidacloprid on sheep keds (Melophagus ovinus): a light and electron microscopic study

The effects of imidacloprid (Advantage) on sheep keds (Melophagus ovinus Linné 1758) were studied in vivo and in vitro by means of direct observation (monitored on video tape) and by light and electron microscopy. It was found that: 1. Imidacloprid acted rapidly on all motile stages of the sheep keds. Within 3–4 min after exposure they became immobile and their legs and the abdomen started tetanic trembling movements for 15–30 min, leading to death. 2. The compound is apparently taken up by the body, since it also acted on those sheep keds that had been exclusively exposed to imidacloprid-contaminated filter papers. 3. The compound is available and active for more than 1 month in the wool of sheep; even rainfall does not reduce its efficacy. Body contact between treated mother sheep and their lambs protects them from infestation with these ectoparasites. 4. The compound initiates an ultimately lethal destruction of the ganglia, nerve chords and related muscle fibers, as can be seen in electron micrographs. Received: 7 October 2000 / Accepted: 18 October 2000     

Plexiform neurilemoma: A clinicopathological and immunohistochemical analysis of 23 tumours from 20 patients

Summary A clinicopathological and immunohistochemical study was done on 23 plexiform (multinodular) neurilemomas excised from 20 Japanese. Ages of the subjects ranged from 2 years to 69 years with a mean age of 30 years. The tumours occurred most often on the trunk (14), and were located commonly in the dermis and subcutis (19). Three lesions apparently originated from the peripheral nerve trunks. Multiple tumours were observed in six instances, and two were associated with von Recklinghausen's disease. Microscopically, they appeared as multinodular growths, most nodules were moderately cellular, and both Antoni A and B patterns were distinct in 10 tumours. Obvious Verocay bodies were noted in seven tumours and abortive ones in five. Immunohistochemical reactivity to S-100 protein was demonstrated in both nuclei and cytoplasm of almost all tumour cells of all lesions examined. Recurrences are nil among the 4 patients who could be followed. Correlations with trauma and with von Recklinghausen's disease are briefly discussed.     

经颅磁刺激安全性的实验研究

目的 :探讨重复经颅磁刺激的安全性。方法 :选用 3 6只健康Wistar大鼠 ,随机分为对照组 ,低频刺激组和高频刺激组 ,对低频组和高频组分别给予不同频率和强度的经颅磁刺激 ,然后对各组大鼠的行为、组织病理形态学、血清髓鞘碱性蛋白 (MBP)及神经元特异性烯醇化酶 (NSE)含量进行观察。结果 :低频刺激组 (5Hz)和高频刺激组 (2 0Hz)在刺激过程中均未出现异常活动 ,无肢体强直、阵挛等 ,脑组织形态学包括大体观察、普通光镜及电镜改变不明显 ,其血清MBP和NSE含量与正常对照组比较 ,差异无显著性意义 (P >0 0 5 )。结论 :在一定强度和频率内经颅磁刺激是一种比较安全的方法     

Apolipoprotein D is down-regulated during malignant transformation of neurofibromas

Apolipoprotein D (apoD) expression was studied in nonneoplastic peripheral nerve, neurofibromas (NFs), and malignant peripheral nerve sheath tumors (MPNSTs) by quantitative polymerase chain reaction, in situ hybridization, and immunohistochemistry. Multiplex quantitative polymerase chain reaction for messenger RNA was performed on a series of formalin-fixed and paraffin-embedded specimens that included 9 MPNSTs, 12 NFs, and 4 normal peripheral nerves. The average apoD expression was 108-fold decreased (DeltaCt = -7.3) in the MPNSTs compared with the NFs (P < .05). ApoD expression levels were 3.0-fold elevated (DeltaCt = 1.7) in the NFs compared with nonneoplastic peripheral nerve (P < .05). In situ hybridization for apoD RNA was performed on a separate series of 10 cases in which each microscopic section included both MPNST and the NF from which it arose. These studies confirmed elevated apoD expression in NFs compared with MPNSTs and demonstrated that this expression was variable among individual cells within the NFs. Differential expression by immunohistochemistry could only be demonstrated in selected areas, most likely because apoD protein is a small molecule that is secreted out of the cell into the extracellular space and plasma. ApoD expression initially increases a small amount with the formation of NFs from nonneoplastic peripheral nerve and subsequently decreases markedly as NFs transform into MPNSTs. This expression pattern may serve as a marker for cell cycle inhibition during peripheral nerve tumorigenesis.     

Creation of a model for multiple sclerosis in Callithrix jacchus marmosets

 Multiple sclerosis (MS) is an inflammatory disease of the CNS white matter characterized pathologically by the accumulation of perivascular and parenchymal T lymphocytes (T cells), and macrophage infiltration associated with myelin destruction. MS lesions are also characterized by the death of oligodendrocytes (the myelin-producing cells) and proliferation and hypertrophy of astrocytes with scar tissue (gliosis) replacing normal myelin. These changes result in the loss of axonal conduction for neurons of the CNS and in clinical disability. MS is thought to be an autoimmune disease, in particular because of its analogy with the disease model of experimental allergic encephalomyelitis (EAE). Despite extensive research and the availability of various EAE models in laboratory rodents the etiology of human MS has not been identified, and to date no effective treatment exists. Phylogenetic differences may limit the usefulness of existing EAE models, and indeed no single form of rodent EAE recapitulates all the clinical and pathological features of MS. Here we describe a novel form of EAE created in a nonhuman primate, the common marmoset Callithrix jacchus. Active immunization of these monkeys with whole myelin produces a primary demyelinating disease with a chronic relapsing-remitting course, characterized pathologically by moderate inflammation with prominent and early demyelination and gliosis reminiscent of human MS. Adoptive and passive transfer experiments have permitted definition of the mechanisms responsible for the MS-like pathology. Production of the fully demyelinated lesion requires synergism between encephalitogenic (e.g., disease-inducing) T cells and pathogenic antibody. The antigens of myelin that promote encephalitogenic T cell and antibody responses in this system have been identified. Because of the similarity between the two conditions and the high degree of conservation in immune and nervous system genes between nonhuman primates and humans, future studies of marmoset EAE will likely accelerate the development of therapies for human MS. Received: 6 June 1996 / Accepted: 17 December 1996     

T cell epitope spreading to myelin oligodendrocyte glycoprotein in HLA-DR4 transgenic mice during experimental autoimmune encephalomyelitis

Epitope spreading has been implicated in the pathogenesis of experimental autoimmune encephalomyelitis (EAE) and human multiple sclerosis (MS). T cell epitope spreading has been demonstrated in rodents for myelin basic protein (MBP) and proteolipid protein (PLP) determinants, but not for myelin oligodendrocyte glycoprotein (MOG), another important myelin antigen. Moreover, the role of human autoimmunity-associated MHC molecules in epitope spreading, including HLA-DR2 and DR4, has not been formally examined. To address these questions, we investigated epitope spreading to MOG determinants in HLA-DR4 (DRB1*0401) transgenic mice during EAE. The data show that upon induction of EAE in HLA-DR4 transgenic mice with the immunodominant HLA-DR4-restricted MOG peptide 97-108 (MOG(97-108); TCFFRDHSYQEE), the T cell response diversifies over time to MOG(181-200) (core: MOG(183-191); FVIVPVLGP) and MBP. The spreading epitope MOG(181-200) binds with high affinity to HLA-DRB1*0401 and is presented by human HLA-DRB1*0401+antigen presenting cells. Moreover, this epitope is encephalitogenic in HLA-DRB1*0401 transgenic mice. This study demonstrates intra- and intermolecular epitope spreading to MOG and MBP in "humanized" HLA-DR4 transgenic mice.     

Reticular and plexiform perineurioma: clinicopathological and immunohistochemical analysis of two cases and review of perineurial neoplasms of skin and soft tissues

Perineurioma represents a recently described and relatively rare neoplasm in the spectrum of benign peripheral nerve sheath tumours composed of perineurial cells staining immunohistochemically positive for epithelial membrane antigen (EMA). In addition to intraneural, extraneural and sclerosing perineurioma, rare variants of perineurioma may occur, and their knowledge is important in the differential diagnosis of mesenchymal tumours of different lines of differentiation and clinically more aggressive neoplasms. We present a case of deep-seated reticular perineurioma arising on the upper arm of a 34-year-old female and a case of a dermal plexiform perineurioma arising on the lower lip of a 60-year-old female. The diagnosis was confirmed in both cases immunohistochemically; neoplastic cells stained positively for EMA and for the newly described perineurial markers, claudin-1 and glut-1. The morphological spectrum and the differential diagnosis of perineurial neoplasms of skin and soft tissues are discussed.     

Ultrastructure of the neuroglial fatty metamorphosis (virchow) in the perinatal period

Summary The ultrastructure of neuroglial fatty metamorphosis (GFM) has been investigated in the telencephalic white matter of 12 premature and mature infants (gestational age 22–40 weeks; survival 0–96 days). GFM was found in all cases apart from a 22-week-old fetus, and involves predominantly astrocytic cells (68.8%), then glioblasts (43.5%), but only 7.4% of oligodendrocytes. GFM, therefore, seems to be independent of the myelination process and indicates the vulnerability of the immature neuroglial population in the metabolic and circulatory disorders of the perinatal period. Since GFM is found in almost all children dying within the early postnatal period, this subtle alteration reflects a special form of minimal brain damage. The relationship between GFM, astrocytic hypertrophy and periventricular leucomalacia and their role in the telencephalic leucoencephalopathy are discussed.

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Zusammenfassung Die Gliazellverfettung im unreifen Großhirn-Marklager wurde bei 12 Kindern ultrastrukturell untersucht (Gestationsalter 22–40 Wochen; Überlebenszeit 0–96 Tage). Die fettige Metamorphose der Neuroglia (Virchow) fand sich in allen Fällen, ausgenommen den 22 Wochen alten Feten, und betrifft vorwiegend junge Astrozyten (68,8%), ferner zu 43,5% unreife Vorstufen, jedoch nur zu 7.4% die (z.Z. der Geburt erst in Erscheinung tretende) Oligodendroglia. Die Fett-Metamorphose der unreifen Glia stellt einen sensiblen Indikator für metabolisch-zirkulatorische Störungen der Perinatalperiode dar und erfolgt unabhängig von dem Prozeß der Markscheidenbildung. Zusammen mit einer oft auffälligen Astroglia-Proliferation ist die intracytoplasmatische Akkumulation nicht membrangebundener Lipide Ausdruck einer temporären Differenzierungsstörung der unreifen Neuroglia. Die resultierende Reifungsdissoziation mit Unterdrückung der oligodendrozytären Zellinie führt zur retardierten Markscheidenbildung und dem Bild der telencephalen Leucoencephalopathie.