血清烯醇化酶定量检测对视网膜母细胞瘤诊断和监测的应用价值

目的探讨血清烯醇化酶,尤其是神经元特异性烯醇化酶（ｎｅｕｒｏｎ－ｓｐｅｃｉｆｉｃｅｎｏｌａｓｅ,ＮＳＥ）含量测定对视网膜母细胞瘤（ｒｅｔｉｎｏｂｌａｓｔｏｍａ,Ｒｂ）诊断和病情监测的价值。方法应用醋酸纤维薄膜快速电泳＋荧光分光光度法,对３５例经病理检查确诊的Ｒｂ患者,分别于眼球摘除时,术后３、６、１２和≥１８个月后及３１例对照组进行血清烯醇化酶总量和ＮＳＥ含量定量检测,观察比较血清烯醇化酶总量和ＮＳＥ含量的变化。结果Ｒｂ患者血清烯醇化酶总量为２０７．０±６５．９Ｕ／Ｌ,ＮＳＥ含量为９８．４±４０．３Ｕ／Ｌ,与对照组血清测定值（分别为１４０．７±４６．０Ｕ／Ｌ和４１．０±１０．９Ｕ／Ｌ）比较明显升高,两组间差异有显著性（Ｐ＜０．００１）。８２．６％Ｒｂ患者血清ＮＳＥ含量明显高于对照组上限水平。Ｒｂ患儿术后６、１２和≥１８个月血清烯醇化酶总量和ＮＳＥ含量逐渐下降（ＮＳＥ含量下降较缓慢）,与治疗前的测定值相比差异有显著性;至≥１８个月后血清ＮＳＥ含量和烯醇化酶总量降至对照组测定值水平。结论血清烯醇化酶尤其ＮＳＥ含量测定对Ｒｂ患者有一定的辅助诊断价值。     

Clinical report and biochemical analysis of a patient with fumarate hydratase deficiency

Fumarate hydratase deficiency (FHD) is a rare metabolic disease caused by two defective copies of the FH gene, which encodes the Krebs cycle enzyme fumarase. FHD is associated with brain and developmental abnormalities, seizures, and high childhood mortality. We describe the symptoms and treatment of a patient with FHD. While infantile spasms are common in FHD, the patient presented with epileptic spasms later in childhood. Also unexpectedly, the patient responded excellently to lacosamide for her non‐convulsive status epilepticus and epileptic spasms after three first‐line medication trials failed. We biochemically analyzed the patient's two fumarase variants (E432Kfs*17 and D65G). While E432Kfs*17 was extremely enzymatically defective, D65G exhibited only a mild defect, possibly playing a role in the patient's longer survival.     

A novel mutation of the fumarase gene in a family with autosomal recessive fumarase deficiency

Fumarase hydratase (FH) deficiency is a rare familial disorder of the tricarboxylic acid cycle which is characterized by severe neurological impairment in early childhood. Several autosomal recessive mutations in the fumarate hydratase gene have been identified as a cause of the lack of fumarase activity in affected individuals. We describe a novel mutation in nucleotide 1127A>C of the fumarase cDNA which changes glutamine 376 to proline in the vicinity of the catalytic site and explains the loss of FH function. Two homozygous carriers of this mutation suffered from severe encephalopthy and died at a young age. Molecular modeling of FH structure shows that the mutation Gln376Pro in the second half of the fumarase sequence disrupts the structure of the active site. Analysis of the FH mutation and the mutant enzyme variant described here provides an explanation for the mechanism of FH deficiency at the molecular level and paves the way for the analysis of other dysfunctional FH variants.Abbreviations FH Fumarate hydratase - PCR Polymerase chain reaction     

青霉胺治疗引起肝豆状核变性神经症状加重

目的 探讨肝豆状核变性(WD)患者在使用青霉胺治疗过程中出现神经症状加重的原因和机制.方法 选取初诊WD患者40例(其中脑型32例,肝型8例),健康对照20名.对WD患者进行神经症状评分,测定脑脊液铜、血清铜、24 h尿铜、脑脊液和血清神经元特异性烯醇化酶(NSE)、血脑屏障指数(AR值).使用青霉胺治疗3个月后,再次进行神经症状评分,用相同方法测定以上指标.所有数据进行统计学分析.结果 15例(46.9%)脑型WD患者使用青霉胺治疗后出现神经症状加重.症状加重的WI)患者治疗后脑脊液铜[(0.083±0.030)mg/L]、脑脊液及血清中NSE[(3.00±1.17)、(6.79±2.20)μg/L]高于治疗前[分别为(0.072±0.027)mg/L,(1.32±0.85)、(3.45±2.09)μg/L,t=3.12、2.53、2.71,P<0.05],血清铜低于治疗前[治疗前后分别为(0.37±0.09)和(0.25±0.08)mg/L,t=3.17,P<0.05].症状加重的患者尿铜排出量显著低于症状改善者.治疗前后AR值[分别为(9.53±3.18)和(12.24±3.17)]差异无统计学意义(t=1.45,P>0.05).结论 初诊WD患者使用青霉胺后是否出现症状加重与其神经系统病变程度和使用青霉胺治疗的剂量有关.青霉胺治疗早期神经症状加重的原因可能是脑铜的增加、神经元新近的损伤和坏死以及铜从中枢神经系统排出困难.     

孕鼠血清高胆酸水平对胎鼠大脑组织形态的影响

目的 探讨孕鼠血清高胆酸水平对胎鼠大脑组织形态的影响.方法将30只清洁级成年SD孕鼠随机分为A、B、C 3组,每组10只.在妊娠第13～20天,A组孕鼠每天1次腹腔注射纯胆酸5.5 mg·kg-1·d-1(高胆酸);B组孕鼠每天1次腹腔注射纯胆酸1.4 mg·kg-1·d-1(低胆酸);C组同步注射等量生理盐水.妊娠第21天剖腹取胎,测定胎鼠体重及记录死亡率.测定孕鼠和胎鼠血清总胆酸水平,采用酶联免疫吸附试验测定血神经元特异烯醇化酶(NSE)水平.同时,胎鼠断头取脑,采用光镜和电镜观察胎鼠大脑组织的病理改变.结果 (1)3组孕鼠及胎鼠血清胆酸水平分别为:A组(22.3±8.1)μmol/L,(28.8±8.1)μmol/L;B组(9.8±3.6)μmol/L,(9.3±3.5)μmol/L;C组(3.6±1.8)μmol/L,(4.0±1.2)μmol/L.3组间相互比较,差异有统计学意义(P＜0.01).孕鼠血清总胆酸水平与胎鼠血清总胆酸水平呈正相关关系(r=0.875,P＜0.01).(2)胎鼠死亡率在A、B、C组中分别是30.1%,16.9%和7.1%,3组间相互比较,差异有统计学意义(P＜0.05).(3)A、B、C组胎鼠血清NSE水平分别为(31.9±13.1),(13.9±5.9)和(9.3±3.9)ng/L,A组明显高于B组及C组(P＜0.01);B组与C组比较,差异无统计学意义(P＞0.05).胎鼠血清NSE水平与总胆酸水平呈正相关(r=0.758,P＜0.01).(4)胎鼠大脑皮层组织光镜下病理改变:A、B组胎鼠均表现为不同程度的大脑皮层组织层次紊乱,神经元变性坏死,密度降低,胞核固缩深染.胎鼠神经元变性面积积分在A、B、C组中分别是(1.4±0.6),(1.5±0.7)和(0.7±0.3),A组和B组明显高于C组(P＜0.01);A组与B组比较,差异无统计学意义(P＞0.05).胎鼠神经元变性面积积分与胎鼠血清NSE水平的r为0.282(P＞0.05).胎鼠神经元坏死面积积分在A、B、C组中分别是(1.8±0.7),(0.9±0.4)和(0.6±0.3),A组明显高于B组和C组(P＜0.05);B组与C组比较,差异无统计学意义(P＞0.05).胎鼠神经元坏死面积积分与胎鼠血清NSE水平呈正相关关系(r=0.798,P＜0.01).(5)胎鼠神经细胞透射电镜下病理改变:A、B组胎鼠均表现为不同程度的神经细胞核膜双层结构模糊,核固缩及核仁消失,染色质浓缩;内质网和线粒体数目减少,残留线粒体肿胀破坏,嵴空化消失.胎鼠神经细胞内线粒体数密度(Nv)在A、B、C组中分别是(21.9±9.0),(45.5±13.1)和(36.1±12.1)μm-3.A组明显低于B组和C组(P＜0.01);B组与C组比较,差异无统计学意义(P＞0.05).胎鼠神经细胞内线粒体平均体积((V))在A、B、C组中分别是(7.0±1.8),(5.7±1.6)和(3.2±1.2)×10-4μm3.A组和B组明显高于C组(P＜0.01);A组与B组比较,差异无统计学意义(P＞0.05).结论 孕鼠血清中胆酸可使胎鼠脑组织出现明显的病理改变,低水平胆酸主要导致神经细胞变性和线粒体体积增大,而高水平胆酸主要导致神经细胞坏死和线粒体数目减少.胎鼠血清NSE水平与总胆酸水平呈正相关关系.     

茶氨酸对家兔脑缺血后IL-8和NSE及脑超微结构的影响

目的观察茶氨酸对脑缺血后IL-8、NSE和脑超微结构的影响。方法将30只健康家兔随机分为假手术组、脑缺血组和茶氨酸组。假手术组只手术不结扎动脉不使用药物、脑缺血组结扎动脉不使用药物、茶氨酸组结扎动脉并在再灌注时应用茶氨酸。全部动物于缺血前（I）、再灌注时（R0）、再灌注1h（R1）、4h（t12）、12h（R3）、24h（1／4）静脉采血进行白介素-8（IL-8）和神经特异性烯醇化酶（NSE）测定，全部采血完毕后每组取一只动物开颅取3～5块约1mm^3大脑皮层组织进行超微结构观察。结果茶氨酸组脑超微结构改变以及血清IL-8和NSE含量变化均明显轻于脑缺血组。结论提示茶氨酸具有神经保护作用，且无毒副作用，有进一步研究和开发价值。     

Free Radical Oxidation and Catalytic Activity of Aconitate Hydratase in Rat Liver under Normal Conditions and during Toxic Hepatitis

We observed intensification of free radical oxidation, decrease in activity, and changes in catalytic properties of aconitate hydratase in the liver of rats with toxic hepatitis. The total yield and maximum flash intensity of biochemiluminescence increased by 2.2 and 1.7 times, respectively. Differences were revealed in the regulation of aconitate hydratase activity with Fe(2+), Ca(2+), H(2)O(2), and oxidized and reduced glutathione in the liver of rats with toxic hepatitis and control animals.     

Mutations in the AUH gene cause 3-methylglutaconic aciduria type I

The conversion of 3-methylglutaconyl-CoA to 3-hydroxy-3-methylglutaryl-CoA is the only step in leucine catametabolism yet to be characterized at enzyme and DNA levels. The deficiency of the putative mitochondrial enzyme 3-methylglutaconyl-CoA hydratase associates with the rare organic aciduria 3-methylglutaconic aciduria type I (MGA1), but neither the enzyme nor its gene have been described in any organism. Here we report that human 3-methylglutaconyl-CoA hydratase is identical with a previously described RNA-binding protein (designated AUH) possessing enoyl-CoA hydratase activity. Molecular analyses in five patients from four independent families revealed homozygosity or compound heterozygosity for mutations in the AUH gene; most mutations are predicted to completely abolish protein function. Mutations identified include c.80delG, R197X, IVS8-1G>A, A240V, and c.613_614insA. Clinical severity of MGA1 in published patients has been quite variable. Included in the present study is an additional patient with MGA1 who was detected by neonatal screening and has remained asymptomatic up to his present age of 2 years. The boy is homozygous for an N-terminal frameshift mutation in the AUH gene. Complete absence of 3-methylglutaconyl-CoA hydratase/AUH appears to be compatible with normal development in some cases. Further work is required to identify external or genetic factors associated with development of clinical problems in patients with MGA1.     

血清NSE表达水平对肺癌的诊断及治疗价值

目的：探讨血清肿瘤标志物神经元特异性烯醇化酶（NSE）对肺癌的诊断、评估化疗敏感性及病情监测的临床价值。 方法：采用酶联免疫分析方法检测96例肺癌患者、60例肺部良性疾病患者和60例健康对照组血清NSE水平。 结果：肺癌患者血清NSE水平显著高于肺部良性疾病患者及健康人（P<0.001）,其中小细胞肺癌（SCLC）患者NSE水平显著高于肺腺癌及肺鳞癌（P<0.001）。SCLC中扩展性病变血清NSE水平明显高于局限性病变,而非小细胞肺癌（NSCLC）组血清NSE水平同NSCLC临床分期无关联性（P>0.05）。NSCLC组和SCLC组NSE阳性者肺癌近期化疗有效率(66.67% 和87.10%)高于NSE阴性者(33.33%和40.00%)(P<0.05)。 结论：血清NSE是一种有效的肺癌肿瘤标志物。     

浅低温体外循环心内直视手术对病人颅内血浆S-100蛋白和NSE水平的影响

目的 观察浅低温体外循环（CPB）心内直视手术中颅内静脉血中S－100蛋白和NSE的含量变化。方法 择期心内直视手术病人15例,于CPB开始前（A点）,鼻咽温（NPT）降温稳定期（B点）,复温至NPT 36℃（C点）,CPB结束后30min（D点）,CPB结束后4－6h(E点）,CPB结束后24h（F点）经左颈内静脉逆向置管于颈内静脉球部,取血样检测S－100蛋白和NSE的含量。结果 CPB开始后病人颅内静脉血中S－100蛋白和NSE含量显著增加（P＜0．01）,在复温期或升停机后30min达到峰值,以后逐渐下降,停机后24h S－100蛋白和NSE已接近正常水平。结论 CPB可引起S－100蛋白和NSE的释放,提示有脑损伤发生;CPB后24h血浆S－100蛋白和NSE浓度未有显著升高者则无明显神经系统并发症发生。