PGP9．5及S—100蛋白在先天性巨结肠中的表达

目的：观察神经元和神经胶质细胞标志物PGP9．5和S－100蛋白在先天性巨结肠（hirschsprung disease,HD)中的表达。方法：采用PAP免疫组织化学方法。结果（1）在对照组结肠壁神经丛中可见染色深浅不一的PGP9．5免疫反应性神经节细胞,神经纤维均匀分布在肠壁各层,并与肌纤维相平行。神经节细胞胞体对S－100蛋白则表现为细胞状“空白区”。（2）HD结肠壁分化异常,PGP9．5和S－100蛋白免疫反应性神经纤维明显增生,分布紊乱,未见有PGP9．5阳性神经节细胞。在增生的S－100蛋白阳性神经纤维中偶见有细胞状的“空白区”。结论：神经丛中PGP9．5阳性反应的细胞团性和S－100蛋白染色的神经丛中神经丛 “空白区”,可特征性地提示神经节细胞的存在,实验证实,结肠壁神经发育异常是HD的主要病理生理变化。     

神经元特异性烯醇化酶及S-100蛋白对Creutzfeldt-Jakob病诊断价值的研究

目的　探讨早期诊断Creutzfeldt Jakob病 (CJD)的一种简便易行的检测方法。方法　采用ELISA及双抗体夹心方法检测 10例CJD、10例非CJD痴呆患者及 10名健康对照者的血清及CSF中神经元特异性烯醇化酶 (NSE)、S 10 0蛋白水平。同时对CJD患者血清中朊蛋白 (PrP)基因进行检测。结果　CJD组血清及CSF中NSE、S 10 0蛋白均较非CJD痴呆组升高 (均P <0 .0 5 ) ,血清中S 10 0蛋白明显升高 (P <0 .0 1)。CJD组血清中NSE及CSF中NSE、S 10 0蛋白较健康对照组明显升高 (均P <0 .0 1) ,血清中S 10 0蛋白较健康对照组升高(P <0 .0 5 ) ;非CJD痴呆组血清及CSF中NSE、S 10 0蛋白较健康对照组升高 (P <0 .0 5 )。 10例CJD中 8例为12 9密码子甲硫氨酸纯合子型 ,2例 12 9密码子甲硫氨酸和缬氨酸杂合型 ,无 12 9密码子缬氨酸纯合子型。结论　早期CSF中NSE及血清、CSF中S 10 0蛋白检测对CJD可以进行病情的判断及预后的评估。血清中S 10 0蛋白检测可用于CJD与非CJD痴呆的鉴别诊断。散发型CJD大部分为 12 9密码子甲硫氨酸纯合型 ,12 9密码子甲硫氨酸及缬氨酸杂合型CJD其临床表现与纯合型不同。     

Immunohistochemical differential distribution of S-100 alpha and S-100 beta in the peripheral nervous system of the rat

The localization of the alpha subunit of the S-100 protein (S-100 alpha) and beta subunit (S-100 beta) was studied in the peripheral nervous system of the rat. In peripheral nerves, S-100 alpha and S-100 beta were found in the cytoplasm of Schwann cells. Axons were positively stained in part by S-100 alpha and almost totally by S-100 beta. In the dorsal root ganglia, S-100 alpha was found in satellite cells and their processes and in some neurons. S-100 beta was found in more of the large neurons, but almost all of the small neurons were negative for S-100 beta. In the anterior horn cells, S-100 beta staining was stronger than that of S-100 alpha. In Schwann cells, both S-100 alpha and S-100 beta were present on the rough endoplasmic reticulum, free ribosomes, and nucleus, as seen by electron microscopy. The S-100 alpha and S-100 beta in axons were associated with microtubules and neurofilaments.     

S100 protein in folliculostellate cells of the rat pituitary anterior lobe

The anterior lobe of the rat pituitary was investigated immunohistochemicallly as to nervous tissue-specific S100 protein. Granulated cells, both acidophils and basophils, were not stained with peroxidase-labelled anti-S100 rabbit serum (Fab) by the direct staining method. Only stellate-shaped cells with long slender cytoplasmic processes contained nervous tissue-specific S100 protein in the nucleus and cytoplasm, the presence of which was verified by both light and electron microscopic immunohistochemistry. Folliculostellate cells were devoid of specific secretory granules and in areas formed follicles with numerous microvilli and junctional complexes. They often extended long cytoplasmic processes between granulated cells. Ouchterlony double diffusion tests confirmed the presence of S100 protein in the pars distalis of rat pituitary glands.These findings lead to the hypothesis that folliculostellate cells may belong to the neuroectodermal cells and may be specialized cells playing some unknown role in rat adenohypophysis.     

急性脑梗死患者S-100蛋白的动态变化及其临床意义

目的　观察脑梗死患者血浆 S- 10 0蛋白的浓度变化 ,并评价其临床意义。方法　应用 EL ISA法对35例急性脑梗死患者的血浆 S- 10 0蛋白水平进行动态检测 ,同时应用 NIHSS进行神经功能缺损评分及 CT扫描 ,并与 30例对照组患者进行比较。结果　病例组患者 S- 10 0浓度明显升高 ,2～ 3d达到峰值 ,且有严重神经功能缺失的患者 ,S- 10 0升高更明显 ,S- 10 0 >1.0μg/ L、NIHSS 12分提示患者预后不良。结论　缺血性脑梗死后血浆中 S- 10 0蛋白的出现与坏死的神经胶质细胞漏出有关 ,并通过受损的血脑屏障进入血液 ,S- 10 0蛋白可作为缺血性脑损伤尤其是大面积脑梗死早期的外周标志物 ,是比 CT更为敏感的指标 ,对指导治疗有帮助。     

S100B and schizophrenia

The research for peripheral biological markers of schizophrenia, although abundant, has been unfruitful. In the last 2 decades, the S100B protein has made its own room in this area of research. S100B is a calcium‐binding protein that has been proposed as a marker of astrocyte activation and brain dysfunction. Research results on S100B concentrations and schizophrenia clinical diagnosis are very consistent; patients with schizophrenia have higher S100B concentrations than healthy controls. The results regarding schizophrenia subtypes and clinical characteristics are not as conclusive. Age of patients, body mass index, illness duration and age at onset have been found to show no correlation, a positive correlation or a negative correlation with S100B levels. With respect to psychopathology, S100B data are inconclusive. Positive, negative and absence of correlation between S100B concentrations and positive and negative psychopathology have been reported. Methodological biases, such as day/night and seasonal variations, the use of anticoagulants to treat biological samples, the type of analytical technique to measure S100B and the different psychopathological scales to measure schizophrenia symptoms, are some of the factors that should be taken into account when researching into this area in order to reduce the variability of the reported results. The clinical implications of S100B changes in schizophrenia remain to be elucidated.     

Time dependent alterations of co-localization of S100β and GFAP in the MPTP-treated mice

Summary. S100β is a calcium-binding peptide produced by astrocytes. This protein is expressed at high levels in brain and is known as a marker of brain damage. However, little is known about the role of S100β protein during neuronal damage caused by MPTP. To determine exactly changes of expression of S100β protein in relation to changes of glial cells, we investigated immunohistochemically the expression of S100β protein using MPTP-treated mice. The present study showed that tyrosine hydroxylase (TH) immunoreactivity was decreased in the striatum and substantia nigra from 5 h and 1 day after MPTP treatment, respectively. Thereafter, a severe reduction in TH immunoreactivity was observed in the striatum and substantia nigra 1, 3 and 7 days after MPTP treatment. In our double-labeled immunostaining, the number of S100-positive/GFAP-negative cells decreased from 1 day up to 7 days after MPTP treatment. In contrast, the number of double-labeled S100/GFAP-immnoreactive cells increased from 1 day up to 7 days after MPTP treatment. The number of S100β-positive/GFAP-negative cells also decreased 3 and 7 days after MPTP treatment. In contrast, the number of double-labeled S100β/GFAP-immunoreactive cells increased from 1 day up to 7 days after MPTP treatment. The present study demonstrates that S100β/GFAP-positive cells may play some role in the pathogenesis of MPTP-induced dopaminergic neurodegeneration in the striatum. The present results also suggest the presence of the S100β protein in a subpopulation of GFAP-negative astrocytes in the striatum after MPTP treatment. These results suggest that the modulation of astrocytic activation may offer a novel therapeutic strategy of Parkinson’s disease.     

Involvement of S-100 protein in anoxic long-term potentiation

In in vitro rat hippocampal slices a short period (2 min) of hypoxia resulted in lasting potentiation of the population spike transynaptically evoked in CA1 by stimulation of Schaffer collaterals (“anoxic LTP”). Pretreatment of slices with antiserum against S-100 protein fully prevented this anoxic LTP. Since also “classical” (i.e., induced by high-frequency electrical stimulation) long-term potentiation is prevented by anti S-100 serum, this represents one more important similarity between these events.     

中等剂量（20Gy）电离辐照后早期大鼠脑内S—100免疫反应胶质细胞的动态变化

应用大鼠半脑 2 0Gy照射模型和免疫组织化学方法 ,研究电离辐射对脑内S 10 0蛋白表达和变化的影响。于照射后不同时间点 (2 4h ,1,2 ,3,4周 ) ,观察S 10 0蛋白免疫反应胶质细胞在大鼠脑内的分布和数量变化。结果发现 :在上述时间点 ,脑内各部位S 10 0蛋白免疫反应细胞数量进行性增加 ,伴有S 10 0免疫反应胶质细胞的胞体逐渐增大和突起增多 ,与正常对照组具有明显的差异。S 10 0蛋白作为脑内星形胶质细胞功能活化状态的重要指标 ,其表达的显著增加 ,提示脑内星形胶质细胞可能参与了大鼠脑辐射后的早期病理过程。     

Accumulation of ubiquitinated proteins is related to West Nile virus‐induced neuronal apoptosis

West Nile virus (WNV) belongs to the Flaviviridae family of viruses and has emerged as a significant cause of viral encephalitis in humans, animals and birds. It has been reported that WNV replication directly induces neuronal injury, followed by neuronal cell death proven as apoptosis. Therefore, it is important to understand the mechanism of neuronal apoptosis caused by this virus to develop strategies to control its pathogenicity. Accumulation of ubiquitinated abnormal proteins has been reported to be associated with neuronal apoptosis in some pathological conditions. A lot of cellular stresses prevent cellular protein quality control mechanisms, resulting in the accumulation of ubiquitinated abnormal proteins. To obtain a better understanding of the mechanisms of WNV‐induced neuronal apoptosis, we evaluated the accumulation of ubiquitinated proteins in the WNV‐infected neuronal cells. We have observed that WNV infection caused massive neuronal injury in the brains of mice. Viral antigen was detected in the neuronal cytoplasm of the cells exhibiting neuronal apoptosis. Notably, ubiquitinated proteins were detected in WNV‐infected neuronal cells. In addition, accumulation of ubiquitinated proteins was markedly enhanced in mouse neuroblastoma, Neuro‐2a cells after WNV infection. Our histopathological and in vitro studies suggest that accumulation of ubiquitinated proteins in neuronal cells might be associated with neuronal apoptosis caused by WNV infection.     

Distribution of S-100 protein outside the central nervous system

The distribution of S-100 outside the central nervous system in humans and rats was explored using antiserum to S-100 and the peroxidase anti-peroxidase method of Sternberger. In peripheral nerves the Schwann cells and the outermost part of the myelin sheaths were stained; axons were not. In dorsal root ganglia and ganglia of the autonomic nervous system only satellite cells were stained. In the adrenal medulla a considerable number of cells were stained. In all other organs studied Schwann cells and satellite cells of ganglia were the only elements that were stained. We conclude that S-100 could serve as a marker for Schwann cells in situ.     

Reverse cellular distribution of calmodulin to S-100 protein in primate brain

Both calmodulin and S-100 protein are Ca2+-binding proteins of the EF-hand family. Immunocytochemical study revealed that calmodulin existed mainly in the neurons, whereas S-100 protein was localized primarily in the glial cells of the cerebral and cerebellar cortices of man and monkey. The observed reverse cellular distribution of calmodulin to S-100 protein in primate brain suggests that calmodulin might be replaced in its role as a Ca2+-binding protein by S-100 protein in the glial cells.     

Ultrastructural localization of S-100 protein in neurofibroma

Summary The nature of the cells in neurofibromas was studied by electron microscopy and immunoelectron-microscopic examination of S-100 protein. Ultrastructurally, all five neurofibromas studied were found to be composed of Schwann cells, perineurial cells, and intermediate cells, which had features of both perineurial cells and fibroblasts. The Schwann cells had complex, branched cytoplasmic processes and a continuous basal lamina. The perineurial cells were distinguishable from Schwann cells by the presence of numerous pinocytotic vesicles, unbranched slender cytoplasmic processes and a discontinuous basal lamina. The intermediate cells had no basal lamina, but were topographically related to Schwann cells and had a similar fine structure to that of perineurial cells. Thus, they seemed to be modified neoplastic perineurial cells. Immunoelectron-microscopic studies showed the presence of cells with and without S-100 protein in the neurofibromas: cells with S-100 protein resembled Schwann cells ultrastructurally, and those without S-100 protein were perineurial and intermediate cells. Some Schwann cells with S-100 protein in one neurofibroma had numerous pinocytotic vesicles characteristic of perineurial cells, suggesting that Schwann cells and perineurial cells, are functional variants of the same cell type. Thus this study showed that neurofibromas were composed of Schwann cells with S-100 protein and perineurial and intermediate cells, including socalled endoneurial fibroblasts, without S-100 protein. Morphological and functional transition seems to occur between Schwann cells and perineurial cells, and between perineurial cells and intermediate cells.     

Regulation of protein kinases and coregulatory interplay of S-100beta and serotonin transporter on serotonin levels in diabetic rat brain

Protein kinases are critical component in the regulation of signal transduction pathways, including neurotransmitters. Our previous studies have shown that serotonin (5-HT) altered under diabetic condition was accompanied by alterations of protein kinase C-alpha (PKC-alpha) and CaMKII, and those alterations were reversed after insulin administration. The current study showed that alloxan-induced diabetic animals revealed hyperglycemia and was associated with an increase in the content of 5-HT, PKC-alpha expression and PKC activity (P < 0.05) simultaneously in striatum (ST), midbrain (MB), pons medulla (PM), cerebellum (CB), and cerebral cortex (CCX) from 7 days to 60 days. Although the 5-HT levels in hippocampus (HC) and hypothalamus (HT) were not altered, the PKC-alpha expression and PKC activity showed increases (P < 0.05) in level in HC. Insulin administration reversed all these changes to a normal level. In contrast, the in vitro study has shown that the 5-HT levels correlated with PKC-alpha expressions as well as PKC activity (P < 0.05) only in ST, MB, and CB either after induction with phorbol 12-myristate 13-acetate (PMA) or blocking with chelerythrine, whereas PM and CCX remained elevated (P < 0.05), implying a regulatory role for PKC-alpha only in ST, MB, and CB. However, our consecutive studies have shown that the 5-HT level in PM was regulated by p38-mitogen-activated protein kinase (p38-MAPK) both in vivo and in vitro, whereas the 5-HT level in CCX was coregulated by S-100beta by protein-protein interaction with serotonin transporter (SERT) via 8-bromoadenosine 3',5'-cyclic monophosphate sodium salt (8-Br-cAMP)-induced cAMP/PKAII pathway(s).     

S100B蛋白生物学研究与临床应用进展

S100蛋白是一种分子量较小（10～12ku）的EF-手型钙结合蛋白，通过对钙离子的调节及与靶蛋白的相互作用，在体内发挥多种生物学作用．在细胞增殖、分化，肌肉收缩、基因表达、分泌及细胞凋亡中发挥重要作用。1965年Moore等首先在牛脑组织中发现S100蛋白，因其在中性饱和硫酸铵中100％溶解而得名。现已发现S100蛋白家族成员20个，S100B蛋白为其中一员，     

S100B-immunopositive glia is elevated in paranoid as compared to residual schizophrenia: a morphometric study

OBJECTIVE: Several studies have revealed increased S100B levels in peripheral blood and cerebrospinal fluid (CSF) of patients with schizophrenia. In this context, it was postulated that elevated levels of S100B may indicate changes of pathophysiological significance to brain tissue in general and astrocytes in particular. However, no histological study has been published on the cellular distribution of S100B in the brain of individuals with schizophrenia to clarify this hypothesis. METHODS: The cell-density of S100B-immunopositive glia was analyzed in the anterior cingulate, dorsolateral prefrontal (DLPF), orbitofrontal, and superior temporal cortices/adjacent white matter, pyramidal layer/alveus of the hippocampus, and the mediodorsal thalamic nucleus of 18 patients with schizophrenia and 16 matched control subjects. RESULTS: Cortical brain regions contained more S100B-immunopositive glia in the schizophrenia group relative to controls (P=0.046). This effect was caused by the paranoid schizophrenia subgroup (P=0.018). Separate analysis of white matter revealed no diagnostic main group effect (P=0.846). However, the white matter of patients with paranoid schizophrenia contained more (mainly oligodendrocytic) S100B-positive glia as compared to residual schizophrenia (P=0.021). These effects were particularly pronounced in the DLPF brain area. CONCLUSION: Our study reveals distinct histological patterns of S100B immunoeactive glia in two schizophrenia subtypes. This may be indicative of a heterogenic pathophysiology or distinct compensatory abilities: Astro-/oligodendroglial activation may result in increased cellular S100B in paranoid schizophrenia. On the contrary, residual schizophrenia may be caused by white matter oligodendroglial damage or dysfunction, associated with a release of S100B into body fluids.     

S100B, a neurotropic protein that modulates neuronal protein phosphorylation, is upregulated during lesion-induced collateral sprouting and reactive synaptogenesis

Using light and electron microscopic immunocytochemistry, we examined the expression of the Ca²⁺-binding protein S100B in the dentate gyrus of adult rats during lesion-induced sprouting and reactive synaptogenesis. Nine days following unilateral lesioning of the entorhinal cortex, S100B was upregulated in cells primarily in the outer part of the molecular layer of the ipsilateral dentate gyrus. When examined with electron microscopy, numerous astrocytes and synapses containing S100B were identified. These data show that during lesion-induced sprouting and reactive synaptogenesis, S100B is upregulated in astrocytes and can be found in pre- and post-synaptic compartments where it might influence neuronal protein phosphorylation.     

Synergistic interactions between COMT-/MAO-inhibitors and L-Dopa in MPTP-treated mice

Summary Four experiments were performed to investigate the anti-akinesia effects of combining a sub-threshold dose (5mg/kg, s.c.) of L-Dopa with different doses and combinations of COMT and MAO inhibitors upon the hypokinesia observed in MPTP-treated mice. Ro 40-7592 (1 and 3 mg/kg, s.c.), a novel COMT inhibitor, 60 min before L-Dopa reinstated both locomotion and rearing during a 2-hr interval after L-Dopa in MPTP mice; control mice were unaffected. The combination of Ro 40-7592 (3 mg/kg, s.c.) and pargyline (5 mg/kg, s.c.), a MAO inhibitor, with L-Dopa produced increases in both the peak effect and duration of action indicating a distinct potentiation of the effects of Ro 40-7592 by pargyline. L-Deprenyl, a MAO_B inhibitor, together with L-Dopa, restored locomotion and rearing behaviour at all three doses applied (1, 3 and 10 mg/kg, s.c.); in control mice, motor activity was stimulated at the higher doses (3 and 10 mg/kg, s.c.), independent of L-Dopa administration. Combining L-Deprenyl (3 mg/kg, s.c.) with Ro 40-7592 (3 mg/kg, s.c.) one hr before L-Dopa to MPTP mice potentiated the restorative effects of each compound by itself, although no increase in peak effect was obtained. In the control mice, L-Deprenyl plus Ro 40-7592 or L-Deprenyl, by itself, stimulated motor activity following injection of L-Dopa. Marked dopamine (DA) depletions in the striatum of MPTP-treated mice were evident. The present results demonstrate that the effects of the COMT/MAO inhibitors in combination, and in conjunction with L-Dopa (at a dose that was without effect by itself), were well in excess of a summation of their individual effects. It was concluded therefore that a synergism of the restorative, anti-akinesic action of these compounds in MPTP-treated mice could offer a broader therapeutic spectrum in the treatment of Parkinson's disorder.     

高血压脑出血患者血清S-100蛋白的表达及手术对其影响

目的探讨高血压脑出血患者血清S-100蛋白的表达及手术治疗对其影响和临床意义。方法选择出血量30～50ml的高血压脑出血患者67例，分为手术组35例和保守组32例。用酶联免疫吸附测定法（ELISA）动态测定血清中S-100蛋白的含量。结果①保守组患者S-100蛋白浓度在发病21d内高于正常组（P〈0．01）；②手术组患者S-100蛋白浓度在发病7d内高于正常组（P〈0．01），15d和21d时与正常组比较无显著性差异（P〉0．05）；③手术组患者S-100蛋白浓度在发病3d内与保守组比较无显著性差异（P〉0．05），但下降较快，在7d时低于保守组，在15d和21d时明显低于保守组（P〈0．001）。结论手术清除血肿可以减轻高血压脑出血患者的脑损伤。     

Poly(ADP‐ribose)polymerase inhibitor can attenuate the neuronal death after 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine‐induced neurotoxicity in mice

An excessive expression of poly(ADP‐ribose)polymerase (PARP) has been demonstrated to play a key role in the pathogenesis of Parkinson's disease (PD). Here we investigated the therapeutic effect of the PARP inhibitor benzamide against 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) neurotoxicity in mice. In our HPLC and Western blot analysis, pretreatment with benzamide showed a neuroprotective effect against MPTP neurotoxicity in mice. Posttreatment with benzamide also attenuated MPTP neurotoxicity in mice. Furthermore, our immunohistochemical study showed that posttreatment with benzamide significantly prevented neuronal damage by suppressing overexpression of neuronal, microglial, and astroglial PARP after MPTP treatment. These findings have important implications for the therapeutic time window and choice of PARP inhibitors in PD patients. Our present findings provide further evidence that PARP inhibitor may offer a novel therapeutic strategy for PD. © 2009 Wiley‐Liss, Inc.