Nitric oxide synthase gene polymorphisms in children with minimal change nephrotic syndrome

Aims:  Nitric oxide (NO) attenuates many functions within the kidney, and all NO synthase (NOS) isoforms are constitutively expressed in the kidney. But the exact role of NO in renal diseases is still debatable. The aim of the present study was to investigate endothelial ( eNOS ), and neuronal ( nNOS ) NOS gene polymorphisms in children with minimal change nephrotic syndrome (MCNS).
Materials and methods:  Eighty-six Turkish children with clinical MCNS, ranging in age from 2 to 10 years, were compared with 114 healthy age- and sex-matched controls. The glu 298 Asp (G/T) polymorphism of the eNOS, and C276T (C/T) polymorphism of nNOS genes were genotyped using polymerase chain reaction.
Results:  The distribution of GG, TG, and TT genotypes for eNOS was 52%, 33% and 15% in MCNS compared with 61%, 26% and 13% in the controls ( P  > 0.05). The distribution of CC, TC, and TT genotypes for nNOS was 16%, 66% and 18% in MCNS compared with 10%, 43% and 47% in the controls. TT genotype distribution of nNOS was found to be lower in patients ( P  = 0.003). The eNOS and nNOS gene polymorphisms were not associated with gender, positive family history, frequency of relapses, or response to steroid.
Conclusions:  The present study is the first to investigate eNOS and nNOS gene polymorphisms in children with MCNS. The nNOS gene polymorphism may be associated with MCNS in children, but further studies in a larger population with different glomerular diseases are needed to confirm the results.     

儿童阴茎海绵体神经显微解剖和临床意义

目的了解儿童阴茎海绵体神经的解剖走向和分布。方法利用手术显微镜，HE染色和神经性一氧化氮合成酶(nNOS)免疫组织化学染色，观察6具完整儿童男尸阴茎海绵体神经的走向和分布。结果阴茎海绵体神经穿过尿生殖膈下筋膜后，走行于阴茎背神经、海绵体动静脉之下，发出1支分支融合入背神经，其余2～3支海绵体神经分支分布于阴茎海绵体脚部。阴茎海绵体神经组织nNOS免疫组织化学染色存在棕黄色nNOS颗粒。结论阴茎海绵体神经于阴茎背神经、海绵体动静脉之下走行，分成分支融合入背神经和分布于阴茎海绵体脚部。     

AGPS在侵袭性纤维瘤侵袭机制中的研究

目的 探索烷基甘油磷酸合成酶(alkyldihydroxyacetonephosphate synthase,AGPS)在侵袭性纤维瘤(aggressive fibromatosis,AF)侵袭能力中发挥的作用;观察异硫氰酸苄酯(Benzyl isothiocyanate,BITC)对AF细胞AGPS的表达及侵袭的影响.方法 收集30例我院病理确诊为AF的标本组织,其中男18例,女12例,患儿平均年龄6岁,原发13例,复发17例,臀部21例,大腿7例,前臂2例,瘤体平均大小336cm3,收集6例来源于内固定取出术的瘢痕组织,提取培养两例原代肿瘤细胞(AF1及AF2),在组织学水平以瘢痕组织为对照通过免疫组化方法,细胞水平以人正常成纤维细胞(HFF)为对照,通过Western blot方法分别观察AGPS在组织及细胞水平的表达;运用SiR-NA干扰技术抑制AF细胞AGPS的表达后采用Wound healing及Transwell观察AF细胞迁移及侵袭能力的改变;5 μm/L BITC干预AF细胞,等量二甲基亚砜(DMSO)作为对照,Western blot测定AF细胞AGPS的表达、Wound healing及Transwell观察AF细胞迁移及侵袭能力的改变.结果 AF组织及细胞中AGPS的表达明显高于对照组(P组织=0.001、PAF1=0.006和PAF2=0.003);SiR-NA沉默AGPS表达后AF细胞的迁移(PWound healing＜0.01,PMigration=0.000)以及侵袭(P=0.000)能力明显减弱;BITC可以降低AGPS的表达(P=0.029),并抑制AF细胞的迁移(Wound healing P值分别为0.039(12 h)及0.004(23 h),migration P=0.000)以及侵袭能力(P=0.000).结论 AGPS在AF的侵袭力中发挥一定的作用,为治疗AF提供新的药物靶点.     

癫癎发作过程中一氧化氮合酶与细胞凋亡的关系

目的探讨癫癎发作过程中一氧化氮合酶(NOS)的变化与细胞凋亡的关系。方法采用戊四氮(PTZ)致癎大鼠模型,利用流式细胞仪(FCM)检测癫癎发作后不同时间点海马神经细胞凋亡情况,比色法检测NOS水平。结果NOS在1、24h有2个分泌高峰,与对照组比较有显著性差异(Pa<0.05);细胞凋亡出现时间稍晚,在6h左右开始升高,至24h达到高峰,48h后有所下降,与对照组比较有显著性差异(Pa<0.05)。结论PTZ致癎大鼠模型NOS早期可能参与癫癎发生和脑保护,后期可能参与神经细胞凋亡。     

地塞米松对小鼠星形胶质细胞诱导型一氧化氮合酶表达的影响

目的 观察内毒素导对体外培养的大鼠星形胶质细胞一氧化氮（ＮＯ）及其诱导一氧化氮合酶（ｉＮＯＳ）ｍＲＮＡ表达的变化,以及地塞米松（ＤＥＸ）对其的影响。方法 采用Ｒｒｉｅｓｓ法和半定量反录－聚合酶链反应（ＲＴ－ＰＣＲ）法,对正常、内毒素诱导后及加入ＤＥＸ的内毒素诱导后大鼠星形交质细胞ＮＯ及其ｉＮＯＳｍＲＮＡ表达进行检测。结果（１）内毒素诱生形胶质细胞ＮＯ产生增另,且呈内毒素剂量和时间依赖性;（２）ＤＥ     

脓毒症与线粒体呼吸链酶复合物Ⅴ研究进展

脓毒症是一种复杂的病理生理功能紊乱性疾病,是由病原微生物感染机体导致的全身性炎性反应,是ICU重症患者的主要死因之一.脓毒症患者发病过程中存在线粒体功能失调以及氧化磷酸化功能受损.线粒体呼吸链酶复合物Ⅴ是线粒体氧化磷酸化的关键酶,在脓毒症前期主要促进ATP合成且酶活性降低;在脓毒症后期即局部缺血的条件下,主要发挥水解功能且活性上调.该文就脓毒症患者复合物Ⅴ活性变化、变化机制以及治疗的研究进展进行阐述.     

Inducible and endothelial constitutive nitric oxide synthase gene polymorphisms in Kawasaki disease

BACKGROUND: Nitric oxide (NO) is secreted by immune and vascular endothelial cells, and appears to play important roles in the pathophysiology of Kawasaki disease (KD). Thus, genetic variations in NO synthase (NOS) genes may be involved in the development of coronary artery lesions (CAL) in KD. METHODS: The present study investigated the association of endothelial constitutive NOS (ecNOS) and inducible NOS (iNOS) gene polymorphisms with the development of CAL in KD in a Japanese population. RESULTS: The genotype distributions of 27-bp tandem repeat polymorphism within intron 4 of ecNOS gene did not show any significant difference between controls and KD patients with or without CAL. In addition, there was no significant association between whole-allele distribution of iNOS gene promoter (penta-repeat CCTTT) polymorphism and KD with or without CAL. CONCLUSION: These results did not support any association of ecNOS and iNOS gene polymorphisms to the development of CAL in KD patients in a Japanese population.     

Urinary homocystine levels in a newborn infant with cystathionine synthase deficiency

A boy with homocystinuria due to cystathionine synthase deficiency was found to have hypermethioninaemia by neonatal blood screening, but was not diagnosed as homocystinuric until 3 months of age because urinary homocystine was not detected by the cyanidenitroprusside test or on two examinations with a sensitive amino acid autoanalyser. These findings indicate that tests for urinary homocystine should be made repeatedly with an amino acid auto-analyser in newborn infants with hypermethioninaemia until the enzyme defect is identified.     

Snyder-Robinson综合征1例报告及文献复习

<正>Snyder-Robinson综合征（SRS）是一种X连锁智力障碍性疾病,由精胺合酶（SMS）基因突变引起,主要表现为智力障碍、肌张力低下、骨质疏松等。目前国内尚未见报道。本文报道1例新生儿期SRS的临床表现及基因测序结果,以加强临床医生对本病的认识。1病历资料患儿男,出生后3 h。因“出生后3 h发现面部不对称、不完全腭裂”于2020-07-27入山东第一医科大学第一附属医院新生儿科,     

Hypoglykämien aufgrund eines Glykogensynthasemangels

Summary We describe a patient who was admitted to hospital because of hypoglycemic convulsions at the age of 6 months. Diagnostics revealed that the hypoglycemic episodes were caused by glycogen synthase deficiency. Glycogen synthase deficiency its a rare cause of fasting hypoglycemia in childhood associated with normal production of ketone bodies. It is characterized by supranormal increase of glucose and lactate in blood after a meal or an oral glucose tolerance test. Diagnosis is confirmed by demonstration of a reduced activity of glycogen synthase in liver tissue, and lately also by identification of pathogenic mutations in the glycogensynthase gene (GYS2-gene). Hypoglycemic episodes can be prevented by offering frequent protein-rich meals during the day and a late evening meal with uncooked corn starch. Discussion: Glycogen synthase deficiency is nearly indistinguable from the relatively common ’ketotic hypoglycemia’ and is probably more common than is believed today. Therefore, one should include an oral glucose tolerance test in the diagnostic workup of ’ketotic hypoglycemia’ and if supranormal levels of glucose and lactate occur, molecular examination of the GYS2-gene is recommended.      

Megaloblastic Anemia and Immune Abnormalities in a Patient with Methionine Synthase Deficiency

ABSTRACT. We report a case of methionine synthase deficiency associated with cellular immune deficiency discovered in a 14-year-old boy. Principal findings were: developmental delay, recurrent upper and lower respiratory tract infections, megaloblastic anemia, discovered at 3 months of age, unresponsive to cyanocobalamin and poorly responsive to folinic acid. Biochemical studies showed: an abnormal deoxyuridine suppression test despite normal serum folate, cobalamin and transcobalamin levels; a normal intracellular uptake of these two coenzymes; and an absolute requirement of methionine for fibroblast growth, suggestive of defective methionine synthesis. An absence of methionine synthase activity in the patient's bone marrow and a profound depression of this activity in lymphocytes and liver were found. Hypergammaglobulinemia with variable lymphopenia, depressed lymphocyte transformation after lectin or recall-antigen stimulation, defective delayed-type hypersensitivity and decreased natural killer activity were noted as well. The patient died at the age of 14.     

先天性心脏病合并肺动脉高压时肺组织病理与免疫组化研究

为探究先天性心脏病（先心病）伴肺动脉高压（肺高压，PH）患儿肺组织一氧化氮合酶（eNOS)的表达有否改变，随机选择法洛四联症（tetralogy of Fallot,TOF)、单纯室间隔缺损(ventricular septal defect,VSD)/房间隔缺损（atrial septal defect,ASD)、先心病合并PH患儿48例，分成3组，取少许右肺中叶组织，利用免疫组织化学法对eNOS进行半定量分析。结果显示先心病伴PH组患儿肺组织内皮细胞内的eNOS染色明显弱于TOF组和单纯VSD／ASD组患儿（P＜0．01）。提示先心病PH组肺组织的eNOS含量水平低下，造成内源性NO生成减少，为临床吸入NO治疗PH提供了理论依据。     

持续高氧对新生鼠肺组织一氧化氮、一氧化氮合酶及病理学的动态影响

目的探讨持续吸入高氧后新生大鼠肺组织一氧化氮(NO)含量、一氧化氮合酶(NOS)活力的动态变化及病理学改变。方法足月新生鼠生后12h内分别持续吸入90%±5%的高氧和空气,于1、3、7、14、21d,分别检测肺组织NO含量、NOS活性以及动态观察病理改变。结果(1)NO含量:在7、14和21d,高氧组水平高于空气组,数值分别为(99.38±7.80)vs(88.78±8.00),P<0.05;(128.18±33.78)vs(93.30±16.73),P<0.05;(170.66±34.00)vs(106.37±25.11),P<0.01。(2)NOS活力:在高氧暴露3d时高于空气组,(20.56±2.56)vs(18.25±0.71),P<0.05;并持续至21d,数值分别为(24.09±2.48)vs(21.10±2.38),P<0.05;(25.07±2.06)vs(20.27±4.15),P<0.05;(27.06±4.79)vs(20.45±2.53),P<0.01。(3)病理学:吸高氧3d时炎症反应为主,7d时炎症反应更明显,开始出现肺间隔增宽,肺泡发育受阻,14d和21d间质增生、肺泡化降低越来越明显。结论持续吸入高氧可致新生鼠发生与人类BPD有类似的病理改变;肺组织病理损伤逐渐加重时,肺组织NO含量和NOS活性增加,提示NO可能在BPD中有重要作用。     

结构型一氧化氮合酶基因表达在新生鼠缺氧性脑损伤中的作用

目的　探讨结构型一氧化氮合酶 (cNOS)mRNA表达在新生鼠缺氧性脑损伤发病中的作用。方法　取新生SD大鼠 2 5只随机分组制作模型并行病理鉴定 ;采用RT -PCR测定脑缺氧 1h ,4h和对照组脑组织细胞内cNOSmRNA表达的变化 ,辉度扫描获取数值。结果　鼠缺氧 1h后脑组织中cNOSmRNA表达显著上升 (P <0 .0 1 ) ,4h组较 1h组下降 (P <0 .0 1 ) ,但仍显著高于对照组 (P <0 .0 1 )。结论　脑缺氧早期cNOSmRNA表达增加 ,由其合成的NO对受损脑组织可能起保护作用。     

缺血再灌注大鼠肾脏诱导型一氧化氮合酶表达的意义

目的观察缺血及缺血再灌注(IR)大鼠肾脏诱导型一氧化氮合酶(iNOS)表达变化的规律。方法制作大鼠肾脏缺血及IR模型,不同时间摘取肾脏,免疫组织化学法检测iNOS表达变化。HE染色观察肾脏损伤程度。结果缺血时及IR后iNOS表达均明显增强(P<0.05),且随时间呈一定的变化规律,呈先升高后下降趋势,24 h达峰值,72 h时降至2 h水平。结论肾脏缺血早期iNOS即起损伤作用,并于再灌注后加重损伤。     

正常肌肉和Duchenne肌营养不良肌肉中脑型一氧化氮合酶mRNA及 …

目的 检测正常人与Ｄｕｃｈｅｒｎｎｅ肌营养不良症（ＤＭＤ）患儿肌肉中脑型一氧人 酶（ｎＮＯＳ）ｍＲＮＡ及其蛋白的表达水平。方法 建立高敏感性的ＲＮＡ酶保护实验,并通过Ｗｅｓｔｅｒｎｂｌｏｔ分析的方法,对１０例ＤＭＤ肌肉标本和５例正常儿童肌肉标本中ｎＮＯＲｍＲＮＡ及其蛋白表达情况进行检测。结果 ＤＭＤ患儿肌肉中ｎＮＯＳｍＲＮＡ的表达量只有正常肌肉的１０％、ｎＮＯＳ蛋白亦有相同的表达规律。结论ｎＮＯＲ     

NOS1 polymorphism is associated with atopy but not exhaled nitric oxide levels in healthy children

Exhaled nitric oxide (FE_NO) is raised in atopy. The mechanism for this is unclear. The aim of this study was to investigate whether the number of AAT repeats in intron 20 of the NOS1 gene, recently associated with variations in FE_NO in adults with asthma and cystic fibrosis, was associated with the raised FE_NO in healthy atopic children. Eighty-seven healthy children (44 girls, 42 atopic, age range 6–18 years) underwent measurements of FE_NO, spirometry, airway responsiveness and skin prick testing. Genotyping was carried out to determine the number of AAT repeats. There was no association between the number of AAT repeats and FE_NO in either the whole sample of healthy children (n = 87) or in the subsample of healthy atopics (n = 42). However, a greater number of atopic children had two high repeat alleles compared with non-atopic children (33.3% vs. 13.6%, respectively, p = 0.03). This suggests that variations in the NOS1 gene may contribute to atopy without this relationship being reflected by FE_NO.