鼠神经生长因子联合神经节苷脂对早产儿脑神经功能的调节效果

目的 探讨鼠神经生长因子联合神经节苷脂对早产儿脑神经功能的调节作用。方法 选择2017年6月—2019年6月商洛市中心医院收治的早产儿86例,随机分为两组,每组各43例。对照组静脉滴注单唾液酸四己糖神经节苷脂钠注射液,将20 mg药物混入10%葡萄糖注射液20 mL中静脉滴注,对于病情严重的患儿可增加1～2个疗程。观察组在对照组治疗的基础上联合使用注射用鼠神经生长因子,每瓶用2 mL氯化钠注射液溶解后肌肉注射,1次/d。两组患儿连续治疗10 d为1个疗程,共治疗3个疗程。观察两组患者的临床疗效,比较两组患儿的临床症状恢复时间、神经损伤标志物水平和NBNA评分。结果 治疗后,观察组新生儿临床治疗总有效率为90.70%,明显高于对照组的69.77%（P<0.05）。治疗后,观察组新生儿意识恢复时间、吮吸能力恢复时间、原始反射恢复时间和肌张力恢复时间均明显少于对照组（P<0.05）。治疗后,两组新生儿血清髓鞘碱性蛋白（MBP）、S-100β蛋白水平均明显降低（P<0.05）,观察组降低程度更为显著（P<0.05）。治疗后,两组早产儿NBNA评分逐渐升高且高于治疗前（P<0.05）,观察组改善程度优于对照组（P<0.05）。结论 鼠神经生长因子联合神经节苷脂治疗早产儿后能够明显降低早产儿神经损伤标志物MBP、S-100β水平,缩短神经功能恢复时间,提高NBNA评分,提高临床治疗效果,可推广使用。     

Insights into the immunogenetic basis of two ganglioside-associated idiotypic networks

The heavy-chain variable regions (VH) from 14F7 MAb, an IgG1 antibody specific for GM3(NeuGc) ganglioside, and its anti-idiotype, the 4G9 MAb, were cloned and sequenced. Comparison with previously reported sequences showed that VH 14F7 belongs to the J558(VHI) gene family and that it is highly mutated. VH 4G9 belongs to the Q52(VHII) gene family. The HCDR3 14F7 sequence contains three basic residues that could be involved in the binding to 4G9 MAb, which bears acidic residues in its HCDR3. Studies performed in the syngeneic model showed that 14F7 MAb requires both coupling to KLH and the use of Freund's adjuvant to induce an effective anti-idiotypic IgG (Ab2) response. In contrast, P3 MAb, a germline gene-encoded Ab1 that also recognizes the GM3(NeuGc) ganglioside through a basic motif in its H-CDRs, has been reported to be immunogenic in syngeneic mice, even when injected in saline. In addition, when Leghorn chickens were immunized with 14F7 or P3 MAbs emulsified in Freund's adjuvant, only P3-immunized animals were able to develop antibodies that recognized NeuGc-containing gangliosides, antigens which are not present in the normal tissues of this animal species. This phenomenon could be due to the lack of idiotypic connectivity of 14F7MAb.     

丹参注射液联合神经节苷脂辅助治疗康复期脊髓损伤患者的临床疗效及机制

目的:探讨丹参注射液联合神经节苷脂辅助治疗康复期脊髓损伤(SCI)患者的临床疗效,挖掘其作用机制。方法:将新疆石河子大学第一附属医院收治的113例脊髓损伤患者随机分为对照组(n=25)、丹参注射液组(n=28)、神经节苷脂组(n=30)和联合组(n=30),对照组患者给予正常康复训练,神经节苷脂组在对照组基础上,给予静脉滴注神经节苷脂治疗60mg,每日1次,丹参注射液组在对照组基础上给予静脉滴注丹参注射液治疗10 m L,每日1次,联合组在对照组治疗基础上给予神经节苷脂联合丹参注射液治疗,连续治疗20 d,停药10 d。比较两组患者的治疗总有效率和不良反应发生率,同时于治疗前后,比较各炎症因子水平和运动、痛觉及触觉等神经功能康复指标评分;采用ELISA法检测各组患者治疗前后血清中炎症相关因子TNF-α、IL-8、IL-1β及氧化应激相关因子SOD、MDA、VE的表达。结果:联合组疗效最佳,总有效率为83.33%,高于对照组(60%)、丹参注射液组(75%)及神经节苷脂(66.67%);联合组患者运动、痛觉及触觉的ASIA评分均明显高于对照组、丹参注射液组及神经节苷脂组(P<0.05),且联合组肌力恢复正常时间、住院时间及可下地行走时间均明显低于其他组(P<0.05);所有入组患者治疗后血清炎症相关因子TNF-α、IL-8、IL-1β均低于治疗前,但联合组表达最低(P<0.05);丹参注射液组及联合组患者治疗后血清SOD、VE表达升高,MDA降低(P<0.05)。结论:丹参注射液与神经节苷脂联合应用有助于康复期SCI患者恢复,预防脊髓继发性损害,其作用机制可能与抗炎、抗氧化有关。     

神经节苷脂联合鼠神经生长因子对早产儿脑损伤后血清神经损伤标志物的影响

目的观察神经节苷脂联合鼠神经生长因子对早产儿脑损伤后血清神经损伤标志物的影响。方法选取我院胎龄均<37周的脑损伤患儿78例,随机分为对照组和试验组,每组39例。对照组给予胞二磷胆碱注射液0.125 g,每天1次,静脉滴注;试验组在对照组的基础上给予单唾液酸四己糖神经节苷脂钠注射液20 mg+注射用鼠神经生长因子18μg,每天1次,静脉滴注。2组患儿均治疗14 d。出生24 h内和出生14 d内收集血液标本,测定血清髓鞘碱性蛋白(MBP)、S^-100β蛋白、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和Toll样受体-4(TLR-4)水平,同时比较2组患者的临床疗效和药物不良反应发生情况。结果治疗后,试验组和对照组的总改善率分别为94.87%(37例/39例)和76.92%(30例/39例),差异有统计学意义(P<0.05)。治疗后,试验组和对照组的血清MBP分别为(2.01±0.58)和(2.46±0.67)μg·L^-1,S^-100β分别为(0.69±0.28)和(0.91±0.35)μg·L^-1,血清TNF-α分别为(2.04±1.03)和(3.15±1.26)μg·L^-1,IL-6分别为(161.78±44.18)和(257.63±51.36)ng·L^-1,TLR-4分别为(8.45±4.37)和(11.68±4.94)pg·m L^-1,意识恢复时间分别为(2.30±0.57)和(3.21±0.64)d,吸吮能力恢复时间分别为(4.17±0.42)和(5.01±0.45)d,原始反射恢复时间分别为(4.23±0.48)和(6.85±0.59)d,肌张力恢复时间分别为(4.68±0.49)和(7.04±0.53)d,差异均有统计学意义(均P<0.05)。试验组的药物不良反应主要有胃肠道反应及发热,药物不良反应发生率为10.26%(4例/39例);对照组的药物不良反应主要有胃肠道反应、躁动及发热,药物不良反应发生率为12.82%(5例/39例),差异无统计学意义(P>0.05)。结论神经节苷脂联合鼠神经生长因子对脑损伤早产儿的临床疗效确切,有利于缩短恢复时间,改善神经功能,且药物安全性高。     

人脐带间充质干细胞联用神经节苷酯GM1治疗新生大鼠缺氧缺血性脑损伤

目的 观察人脐带间充质干细胞(human umbilical cord mesenchymal stem cells, hUCMSCs)移植及联用神经节苷酯GM1治疗新生大鼠缺氧缺血性脑损伤(hypoxic-ischemic brain damage, HIBD)的效果。方法 RICE法制备七日龄新生大鼠HIBD模型, 随机分为hUCMSCs移植组(n=15)、hUCMSCs+GM1组(n=15), 72 h后经颈静脉移植等量的hUCMSCs, hUCMSCs+GM1组再腹腔注射GM1, 两组均在移植后第31、32、33、34、35天用Morris水迷宫对两组大鼠行为学测试, 比较两组大鼠脑功能恢复情况。免疫荧光染色观察DiI标记的hUCMSCs移植后在两组大鼠脑中的分布情况;神经元前体细胞标记DCX特异性染色观察hUCMSCs移植后分化成神经元前体细胞的情况。结果 Morris水迷宫测试:hUCMSCs+GM1组逃避潜伏期明显短于hUCMSCs移植组, 差异具有统计学意义(P＜0.05)。移植后hUCMSCs能迁徙到脑组织中, 并分布于缺血区的小血管旁, 能分化成神经元前体细胞。hUCMSCs+GM1组大鼠脑片干细胞分布明显多于hUCMSCs移植组(P＜0.05)。结论 hUCMSCs移植对HIBD新生大鼠脑功能有修复作用, 联用GM1, 有协同作用。     

Ganglioside promotes the bridging of sciatic nerve defects in cryopreserved peripheral nerve allografts

Previous studies have shown that exogenous gangliosides promote nervous system regeneration and synapse formation.In this study,10 mm sciatic nerve segments from New Zealand rabbits were thawed from cryopreservation and were used for the repair of left sciatic nerve defects through allograft bridging.Three days later,1 m L ganglioside solution(1 g/L) was subcutaneously injected into the right hind leg of rabbits.Compared with non-injected rats,muscle wet weight ratio was increased at 2–12 weeks after modeling.The quantity of myelinated fibers in regenerated sciatic nerve,myelin thickness and fiber diameter were elevated at 4–12 weeks after modeling.Sciatic nerve potential amplitude and conduction velocity were raised at 8 and 12 weeks,while conduction latencies were decreased at 12 weeks.Experimental findings indicate that ganglioside can promote the regeneration of sciatic nerve defects after repair with cryopreserved peripheral nerve allografts.     

单唾液酸四己糖神经节苷脂治疗新生儿缺氧缺血性脑病的Meta分析

摘 要 目的：系统评价单唾液酸四己糖神经节苷脂（GM1）治疗新生儿缺氧缺血性脑病（HIE）的疗效及安全性。方法：计算机检索PubMed、Clinical Trials、Cochrane Library、Embase、WanFang Data、CNKI、VIP数据库,搜集GM1治疗HIE的随机对照试验（RCTs）,检索时限均为建库至2018年8月31日。由两位研究者独立进行文献筛选、数据提取和偏倚风险评价后,采用RevMan 5.3软件进行Meta分析。 结果：共纳入14个RCTs,包括1 052名患儿。Meta分析结果显示：与常规治疗比较,加用GM1（单用）治疗HIE可提高有效率[RR=1.35,95%CI（1.21,1.51）,P＜0.01];与常规治疗+其他神经保护药比较,加用GM1（联用）治疗HIE可提高有效率[RR=1.20,95%CI（1.07,1.35）,P＜0.01];无论单用或联用GM1均可提高治疗7 d、10~14 d、28 d的新生儿行为神经评分（NBNA）;与常规治疗或者常规治疗+其他神经保护药物的对照组相比,加用GM1可降低头颅CT检查异常发生率[RR=0.34,95%CI（0.18,0.61）,P＜0.01]和远期后遗症发生率[RR=0.25,95%CI（0.12,0.52）,P＜0.01]。结论：基于现有临床证据,GM1可能是HIE有希望的治疗选择,相对来说安全性良好。受纳入研究数量和质量的影响,研究结果有待进一步完善和充实以及更多高质量RCTs予以证实。     

Association of antibodies to ganglioside complexes and conduction blocks in axonal Guillain‐Barré syndrome presenting as acute motor conduction block neuropathy

A close relationship between acute motor conduction block neuropathy and antibodies against the complex of GM1 and GalNAc‐GD1a has been reported. This study investigates the hypothesis that conduction block at the early phase of axonal Guillain‐Barré syndrome (GBS) is also associated with such ganglioside complexes. Sera were obtained from seven French patients with initial evidence of isolated conduction blocks that resolved or progressed to acute motor axonal neuropathy. Serum IgG to asialo‐GM1 and gangliosides of LM1, GM1, GM1b, GD1a, GalNAc‐GD1a, GD1b, GT1a, GT1b, and GQ1b as well as their complexes were measured. Five of seven patients progressed within the first month of disease to AMAN. One patient had IgG antibodies against the complex of asialo‐GM1 and each of the other ganglioside antigens. Another patient carried IgG antibodies against GM1 complex with GM1b, GD1a, and GT1a as well as asialo‐GM1 complex with GD1a and GT1a. None had IgG antibodies against GM1/GalNAc‐GD1a complex. Six patients had IgG against single antigens GM1, GD1a, GalNAc‐GD1a, GD1b, and asialo‐GM1. In three patients, a reduced reaction against GM1/GalNAc‐GD1a complex was observed. The presence of conduction block in axonal GBS is not always associated with anti‐GM1/GalNAc‐GD1a complex antibodies.     

窖蛋白1在无血清状态下诱导FBJ细胞死亡

目的研究无血清状态下,FBJ-S1细胞较FBJ-LL细胞更易死亡的原因。方法用细胞活性检测法分析细胞的活性,分别用HPTLC法、逆转录PCR法和免疫印迹法检测细胞中GD1a和窖蛋白1(caveolin-1)的含量。结果研究表明,在无血清培养基培养条件下,富含神经节苷脂GM3和GD1a的低转移性的FBJ-S1小鼠骨肉瘤细胞趋向死亡,但对含有几乎相同量GM3而无GD1a表达的高转移性的FBJ-LL细胞的生长却未见明显影响。同时发现:a.FBJ-S1细胞富含窖蛋白1,而FBJ-LL细胞中窖蛋白1表达量则较低;b.利用小干扰基因沉默技术在FBJ-S1细胞中沉默窖蛋白1的表达,结果使细胞存活率升高;c.使用GD1a处理FBJ-LL细胞导致其趋向死亡,通过免疫印迹法检测窖蛋白1发现其蛋白表达量增加。结论在无血清培养条件下,窖蛋白1诱导FBJ细胞的死亡。     

GM-1对大鼠视网膜缺血再灌注损伤的保护作用

目的 探讨单唾液酸神经节苷脂（monosialoganglioside,GM-1）对大鼠视网膜缺血冉灌注损伤后视网膜组织及超微结构的保护作用.方法 健康成年Wistar大鼠75只,随机分为3组：正常组5只、NS组35只,GM-1组35只.正常对照组不加任何处理因素,NS组和GM-1组通过升高眼压造成视网膜缺血60 min,分别于缺血前12 h、1 h及缺血结束时3次腹腔注射NS 3mL/kg或GM-1 3mL/kg（10 mg/mL）,并于再灌注0 h（单纯缺血后）、1 h、6 h、12 h、24 h、3 d和7 d共7个时间点取双眼眼球,每时间点5只,HE染色观察视网膜组织结构并测量视网膜内层平均厚度（mean thickhess of the inner retinal layers,MTIRL）,透射电镜观察视网膜超微结构变化.结果 缺血再灌注后,内层视网膜依次表现出水肿、凋亡、萎缩的损伤过程.GM-1可明显减轻其水肿和萎缩的程度,并减少凋亡的发生.结论 GM-1对视网膜缺血再灌注损伤具有明确的保护作用,可能是其有效治疗药物.