Manganese inhalation by rhesus monkeys is associated with brain regional changes in biomarkers of neurotoxicity.

The purpose of this study was to evaluate biochemical markers of neurotoxicity following subchronic manganese sulfate (MnSO(4)) inhalation. Juvenile rhesus monkeys were exposed to MnSO(4) at 0, 0.06, 0.3, or 1.5 mg Mn/m(3) for 65 days. Glutamine synthetase (GS), glutamate transporters (glutamate transporter-1 [GLT-1] and glutamate/aspartate transporter [GLAST]) and tyrosine hydroxylase (TH) protein levels, metallothionein (MT), GLT-1, GLAST, TH and GS mRNA levels, and total glutathione (GSH) levels were assessed in known targets (caudate, globus pallidus, putamen) as well as the cerebellum, frontal cortex, and olfactory cortex. All MnSO(4)-exposed monkeys had decreased pallidal GS protein, decreased caudate GLT-1 mRNA, decreased pallidal GLAST protein, and increased olfactory cortical TH mRNA levels. Monkeys exposed to MnSO(4) at 0.06 or 0.3 mg Mn/m(3) had significantly increased pallidal mRNA levels for GLT-1, GLAST, and TH. Monkeys exposed to MnSO(4) at > or = 0.3 mg Mn/m(3) had several alterations including decreased frontal cortical MT mRNA, decreased caudate, globus pallidus, olfactory cortex, and cerebellum GLT-1 protein, decreased olfactory cortex and cerebellum GLAST protein, increased cerebellar GLAST mRNA, and decreased pallidal TH protein levels. Lastly, GSH levels were significantly increased in the frontal cortex and decreased in the caudate of monkeys exposed to the 1.5-mg Mn/m(3) compared to the controls. Overall, as in our previous studies, we observed that increased Mn concentrations due to airborne Mn exposure differentially affects biomarkers in each brain region (e.g., GSH was increased in the frontal cortex and decreased in the caudate despite two- to threefold increases in Mn concentrations in these regions).     

视神经挫伤对大鼠视网膜谷氨酰胺合酶和兴奋性氨基酸转运蛋白-2表达的影响

目的通过谷氨酰胺合酶(glutamine synthetase,GS)、兴奋性氨基酸转运蛋白-2(GLT-1)表达的改变,研究视神经挫伤后视网膜谷氨酸代谢变化的机制。方法建立大鼠右眼视神经挫伤模型48只。术后1d、7d、14d以高效液相色谱分析检测大鼠玻璃体谷氨酸浓度;通过免疫组织化学检测大鼠视网膜GS、GLT-1的表达。结果视神经挫伤后1d、7d、14d,大鼠玻璃体谷氨酸浓度升高,与对侧眼比较差异具有统计学意义(P=0.0011、P=0.0000、P=0.0000)。视神经挫伤后1d,GS高表达(P=0.0054);挫伤后7d,GS表达与对侧眼比较差异无统计学意义(P=0.1379);挫伤后14d,GS低表达(P=0.0333)。视神经挫伤后1d、7d,GLT-1的表达与对照组的差异无统计学意义(P=0.1985、0.6537);但挫伤后14d,GLT-1低表达(P=0.0403)。结论视神经挫伤后玻璃体谷氨酸浓度升高,与视网膜GS、GLT-1的表达降低有关。     

复方谷氨酰胺治疗腹泻型肠易激综合征的疗效观察

目的观察复方谷氨酰胺治疗腹泻型肠易激综合征的疗效和安全性。方法采用随机对照临床研究，将符合诊断标准的150例腹泻型肠易激综合征患者随机分为复方谷氨酰胺组和奥替溴胺组，复方谷氨酰胺组给予复方谷氨酰胺3粒，每日3次，餐前口服，疗程4周；奥替溴胺组给予奥替溴胺40mg，每日3次口服，疗程4周。观察用药前后腹痛或腹部不适及腹泻评分的变化。结果复方谷氨酰胺可使腹泻型肠易激综合征患者症状评分明显下降，无不良反应。结论复方谷氨酰胺可明昆缓解腹泻型肠易激综合征患者的腹痛和腹部不适及腹泻症状，安全性好。     

谷氨酰胺转运载体的种类与分布及功能特点

谷氨酰胺只有通过谷氨酰胺载体进入效应细胞内才能被利用。因此，氨酰胺载体的作用十分重要。目前，通过基因分离研究证实谷氨酰胺载体蛋白系统分属于4个基因家族。作者就其种类、组织分布及功能特点进行了描述。     

Cellular energetics and glutathione status in NRK-52E cells: toxicological implications

Cellular energetics and redox status were evaluated in NRK-52E cells, a stable cell line derived from rat proximal tubules. To assess toxicological implications of these properties, susceptibility to apoptosis induced by S-(1,2-dichlorovinyl)-L-cysteine (DCVC), a well-known mitochondrial and renal cytotoxicant, was studied. Cells exhibited high activities of several glutathione (GSH)-dependent enzymes, including gamma-glutamylcysteine synthetase, GSH peroxidase, glutathione disulfide reductase, and GSH S-transferase, but very low activities of gamma-glutamyltransferase and alkaline phosphatase, consistent with a low content of brush-border microvilli. Uptake and total cellular accumulation of [14C]alpha-methylglucose was significantly higher when cells were exposed at the basolateral as compared to the brush-border membrane. Similarly, uptake of GSH was nearly 2-fold higher across the basolateral than the brush-border membrane. High activities of (Na(+)+K(+))-ATPase and malic dehydrogenase, but low activities of other mitochondrial enzymes, respiration, and transport of GSH and dicarboxylates into mitochondria were observed. Examination of mitochondrial density by confocal microscopy, using a fluorescent marker (MitoTracker Orange), indicated that NRK-52E cells contain a much lower content of mitochondria than rat renal proximal tubules in vivo. Incubation of cells with DCVC caused time- and concentration-dependent ATP depletion that was largely dependent on transport and bioactivation, as observed in the rat, on induction of apoptosis, and on morphological damage. Comparison with primary cultures of rat and human proximal tubular cells suggests that the NRK-52E cells are modestly less sensitive to DCVC. In most respects, however, NRK-52E cells exhibited functions similar to those of the rat renal proximal tubule in vivo.     

Tolerance of neonatal rat brain to acute hyperammonemia

The aim of the present work was to study the effects of hyperammonemia on brain energy metabolism in neonatal rats. Rats were rendered hyperammonemic by ammonium acetate administration. This decreased brain ATP concentrations but enhanced brain ammonia and lactate levels in both adult and neonatal rats. In adult rats, the decrease in brain ATP concentrations was accompanied by a plunge in the respiratory control rate (RCR) of brain mitochondria. However, the ammonia-induced effect on RCR was not observed in neonatal rats, suggesting that the fall in ATP levels observed in neonatal rats would not be due to an impairment of mitochondrial respiratory efficiency. However, in neonatal rats the increase in blood and brain ammonia concentrations did not change brain glutamate concentrations but decreased glutamine contents. These results may be of relevance for the understanding of the resistance of neonatal rats observed in this work to acute ammonia toxicity     

Involvement of 5-HT7 receptors in serotonergic effects on spike afterpotentials in presumed jaw-closing motoneurons of rats

Supporting glial cells of the peripheral nervous system include satellite cells of dorsal root ganglia and Schwann cells of peripheral nerves. In the central nervous system, glial cells contain enzymes related to the tricarboxylic acid and glutamine cycles: pyruvate carboxylase, glutamate dehydrogenase, and glutamine synthetase. The present study used immunohistochemistry in the rat peripheral nervous system to determine the cellular distribution of these enzymes along with glutamine. In dorsal root ganglia and peripheral nerves, glutamine and glutamine related enzymes were enriched in satellite and Schwann cells. In the dorsal root ganglia, immunoreactive satellite cells surrounded neurons of all sizes. In peripheral nerve, immunoreactive Schwann cells were most easily observed surrounding large diameter, myelinated axons. These Schwann cells contained immunoreactivity in their cell bodies, nodes of Ranvier, and the rim of cytoplasm outside the myelin sheath. Myelin sheaths were non-immunoreactive. The peripheral glial tricarboxylic and glutamine cycles may be used to produce glutamine for neuronal cell uptake and conversion to glutamate for synaptic transmission. Alternatively, these cycles may function in peripheral glia similar to central nervous system astrocytes for supporting the energy demands of neurons.     

谷氨酰胺对新生鼠坏死性小肠结肠炎的影响

目的 探讨谷氨酰胺(glutamine,Gln)对新生鼠坏死性小肠结肠炎(necrotizing enterocolitis,NEC)肠黏膜修复的影响.方法 新生1日龄Wistar大鼠30只随机分为三组,A组为正常对照组;B组为NEC模型组;C组为NEC模型后灌胃给Gln组(2.0 g/kg·d).建立NEC模型,连续3天给予新生鼠100%二氧化碳5 min,然后再给予100%氧气5 min,放回母鼠身边喂养,第4天断头处死新生鼠,取肠道组织待检.分别取近回盲段2～3 cm肠道组织固定、包埋、切片.HE染色光镜下作病理学检查,应用免疫组化技术检测肠黏膜增殖细胞核抗原(PCNA)表达情况,原位末端标记(TUNEL)法检测肠黏膜细胞凋亡的变化.结果 B组HE染色切片见肠壁有不同程度的损伤,病理评分的中位积分为3分;C组有的肠黏膜正常,有的轻度肠上皮细胞脱落,有的绒毛轻度坏死,病理评分的中位积分为1分.B组PCNA数量均低于A组及C组(P＜0.01).B组的肠黏膜细胞凋亡的数量高于A组及C组(P＜0.01).结论 NEC时,新生鼠肠黏膜受损,增殖减慢,细胞凋亡的数量增加;补充Gln可促进NEC新生鼠肠黏膜隐窝细胞增殖,减少肠黏膜细胞凋亡数量,使肠黏膜修复加快.     

ALL患儿肿瘤细胞的门冬酰胺合成酶活性研究

目的：不同类型急性淋巴细胞性白血病(ALL)对左旋门冬酰胺酶(LAsp)敏感度不完全相同,由于LAsp活性水平与细胞内门冬酰胺合成酶活性负相关,因此研究不同类型ALL患儿白血病细胞内门冬酰胺合成酶活性水平分布情况,有助于临床治疗中合理使用LAsp。方法：通过HPLCFLD及蛋白定量等技术,检测28例ALL患儿(免疫酶型TALL7例,B细胞系列ALL21例)白血病细胞内门冬酰胺合成酶活性水平。结果：TALL患儿肿瘤细胞内AS活性水平较B细胞系列ALL患儿的显著增高(P<0.05),其中TALL患儿肿瘤细胞内AS活性中位水平为每小时9.3nMAsn/mgprotein,B细胞系列ALL患儿为每小时5.2nMAsn/mgprotein。但无论是在B细胞系列ALL患儿还是在TALL患儿中,肿瘤细胞内AS活性存在多态性分布。结论：ALL患儿白血病细胞中门冬酰胺合成酶活性存在多态性分布,总体上TALL比B系ALL白血病细胞中门冬酰胺合成酶活性高。