Objective:To analyze the clinical characteristics and prognostic factors of high-risk neuroblastoma (HR-NB) patients with skeletal metastasis.Methods:The clinical features of 336 newly treated HR-NB patients with skeletal metastases admitted to the Department of Medical Oncology of Beijing Children′s Hospital, Capital Medical University from January 2007 to December 2018 were analyzed retrospectively.Kaplan-Meier method was used for the survival analysis, and Log- Rank test was used for univariate prognosis analysis.The Cox model was used to analyze the multifactorial prognostic analysis. Results:A total of 336 HR-NB patients were recruited, involving 188 males and 148 females with the median age of onset of at 43 (4-148) months.Skeletal metastases affected the viscerocranium (89 cases, 26.5%), neurocranium (193 cases, 57.4%), vertebrae (298 cases, 88.7%), sternum and ribs (183 cases, 54.5%), pelvis (270 cases, 80.4%), upper limbs (182 cases, 54.2%) and lower limbs (240 cases, 71.4%). The 5-year event-free survival (EFS) rate and overall survival (OS) rate were (30.4±2.7)% and (41.3±2.9)%, respectively.Univariate analysis showed a significantly lower 5-year OS rate in skeletal metastatic HR-NB patients with poor prognostic classification, the morphology of neuroblastoma (stroma-poor) and ganglioneuroblastoma (intermixed), high index of mitosis-karyorrhexis index, lactate dehydrogenase≥587 U/L, serum ferritin≥92 μg/L, MYCN amplification and 1p loss of heterozygosity, and metastases in the viscerocranium, neurocranium, vertebrae, sternum and ribs, pelvis, upper limbs and lower limbs (all P<0.05). The 5-year OS rate of HR-NB patients with all 7 regions of skeletal metastases was only (14.2±5.9)%, which was significantly lower than that in patients with a single region metastasis or multi-region metastases[(66.0±10.2)% vs.(43.6±3.4)%, χ2=45.722, P<0.05]. Cox multifactorial analysis showed that MYCN amplification ( HR=4.165, 95% CI: 2.356-7.363) and the viscerocranium metastasis ( HR=2.560, 95% CI: 1.519-4.315) were the independent risk factors affecting the prognosis of HR-NB patients with skeletal metastases (all P<0.05). Conclusions:The prognosis is extremely poor in HR-NB patients with multiple skeletal metastases at the initial diagnosis.The amplification of MYCN and the viscerocranium metastasis are the poor prognostic factors for HR-NB patients with skeletal metastases.

背景:前期研究已证实神经营养因子3-壳聚糖载体可支持神经干细胞的存活和增殖,同时可高效诱导神经干细胞向神经元方向分化.目的:观察神经营养因子3-壳聚糖载体对神经元发育进程、发育各阶段电生理特性及发育成熟神经元亚型的影响.方法:取第3代新生大鼠脊髓神经干细胞,分4组培养:空白对照组加入神经干细胞培养基,壳聚糖组加入含壳聚糖的神经干细胞培养基,NT3组加入含神经营养因子3的神经干细胞培养基,NT3-壳聚糖组加入含神经营养因子3-壳聚糖载体的神经干细胞培养基.利用免疫荧光染色观察神经干细胞发育各阶段标志物表达情况,借助全细胞膜片钳技术评价神经干细胞发育过程中电生理特性的变化情况,利用免疫荧光染色观察神经干细胞分化21 d后中间神经元的亚型.结果 与结论:①Nestin、DCX、Tuj1及MAP2免疫荧光染色显示,神经营养因子3-壳聚糖载体维持了神经干细胞池的稳态,并且通过加速神经母细胞的发育进程来促进神经元发育成熟;②全细胞膜片钳记录发育过程中的细胞发现,营养因子神经营养因子3和神经营养因子3-壳聚糖在发育早期对神经干细胞膜功能以及细胞膜上离子通道的发育成熟具有一定的促进作用,但是仅有神经营养因子3-壳聚糖可将这一优势维持到发育中后期,即分化后7-14 d;③免疫荧光染色显示,神经干细胞分化21 d后,NT3-壳聚糖组成熟神经元可表达运动神经元特异性标记物HB9、V1类型中间神经元FOXP1、V2类型中间神经元特异性标记物LHX3,以及调控机械性痛觉感觉中间神经元的特异性标记物VGLUT3;④结果显示,神经营养因子3-壳聚糖载体促进了神经干细胞向神经母细胞的发育,在发育早期对细胞膜功能及细胞膜上的离子通道发育成熟具有一定的促进作用,可诱导发育成熟的神经元亚型多样化.

背景:成年哺乳动物脊髓室管膜细胞损伤后表现出干/祖细胞特性.目的:利用Nestin和Foxj1转基因小鼠标记室管膜细胞,以便追踪室管膜细胞及其子代在成年小鼠脊髓损伤后的增殖分化命运.方法:对转基因小鼠T8脊髓节段完全切除1 mm脊髓组织,术后1-7 d连续腹腔注射BrdU动态观察室管膜细胞,在脊髓损伤后不同时间点(3,7,14,28,56 d)借助BrdU,GFAP,Tuj1,NeuN免疫荧光染色,观察损伤区边缘室管膜细胞的遗传命运谱.结果 与结论:①未损伤脊髓中,Nestin阳性的室管膜细胞处于静止状态;脊髓损伤后室管膜细胞被激活,第3天时在损伤区周围大量增殖;②在损伤后第28天,约3.3％Nestin阳性的室管膜细胞表达神经元标记物Tuj1;③在损伤后第56天,约25.7％Nestin阳性的室管膜细胞分化为星形胶质细胞并构成胶质瘢痕的核心,参与胶质瘢痕的形成;④通过检测室管膜细胞在脊髓损伤后的时空动态变化、增殖和分化特征,为理解脊髓损伤的病理过程提供理论依据,为脊髓损伤修复提供新的思路.

目的 总结单中心胸部软组织肉瘤患儿的临床特征,治疗及预后.方法 回顾性分析2008年6月—2020年5月31日在首都医科大学附属北京儿童医院诊治的原发于胸部的软组织肉瘤,包括尤文肉瘤、横纹肌肉瘤和非横纹肌肉瘤类软组织肿瘤患儿临床资料,分析其临床病理特征,治疗过程和预后相关因素.结果 53例患儿中,男27例,女26例,中位诊断年龄是92(13-193)个月.胸壁肿瘤24例(45.3％),胸腔内肿瘤29例(54.7％).最常见的病理类型是尤文肉瘤,占54.7％.53例患者中1例诊断后回当地治疗,1例患者化疗1疗程后放弃治疗.52例患儿接受化疗,平均化疗11(1-26)个疗程,46例患儿接受原发瘤灶手术切除,47例患儿接受放疗,平均放疗剂量34.9Gy.系统治疗并随访51例患儿的生存分析显示3年总生存率(OS)是(60.0±7.7)％,其中尤文肉瘤3年OS是(64.3±9.9)％,横纹肌肉瘤3年OS是(27.8±14.8)％,其他肉瘤三年OS是(90.0±9.5)％.接受综合治疗(手术+放疗+化疗)的患儿的预后明显优于单纯手术和或化疗的患儿[3 年 OS:(65.4±8.1)％vs(28.6％±17.1)％,x2=12.33,P＜0.001;3 年 EFS:(55.4±0.81)％vs0.0％,x2=28.99,P＜0.001].单因素分析显示肿瘤部位在胸腔内,病理类型为横纹肌肉瘤,发生远处转移的患儿3年OS明显降低(P＜0.05).原发部位位于胸腔的横纹肌肉瘤患儿病死率100％,尤文肉瘤病死率46.6％,其他肉瘤患儿病死率14.2％,三组有明显统计学差异(2x=11.56,P＜0.01).结论 儿童及青少年胸腔内横纹肌肉瘤预后极差.诊断时伴有远处转移及肿瘤位置发生在胸腔内是影响胸部软组织肉瘤患儿预后的不良因素.

Objective:The safety and efficacy of GD2 monoclonal antibody in the treatment of chinese children with high-risk or relapsed/refractory neuroblastoma (NB) in China were evaluated, to provide some basis for the clinical study design and dose determination of dinutuximab β after its marketing.Methods:Among the 37 children with NB there were 26 male and 11 female; 26 high-risk patients, 5 relapsed patients, and 6 refractory patients. A total of 153 cycles of dinutuximab β immunotherapy were completed. The incidence of adverse reactions and initial efficacy of dinutuximab β were evaluated objectively.Results:The pain scores of CRIES in this group were ≤3, and the major manifestations were limb or abdominal pain. All 37 children had fever, with an average peak of 39.4℃. The rate of grade ≥3 infection was 16%. Three children under 3 years old were diagnosed as severe capillary leakage syndrome. Two cases abandoned subsequent immunotherapy after 2 cycles of dinutuximab β treatment due to progressive disease or repeated intestinal obstruction. Up to the last follow-up time, the prognosis of 6 patients was improved ,but 2 cases experienced progressive disease and 2 patients relapsed.Conclusions:Dinutuximab β is well tolerated in high-risk and relapsing/refractory neuroblastoma patients in China. The overall incidence and severity of adverse effect are lower than those reported in foreign literature, but more attention should be paid to the safety of young children under 3 years old.

背景 伴有MYCN基因扩增的神经母细胞瘤(NB)患儿的长期生存率不容乐观,目前国内相关大宗病例报道不多.目的 总结伴有MYCN基因扩增的NB患儿的临床特征、治疗效果及预后相关因素.设计病例系列报告.方法 纳入2007年2月1日至2020年1月30日首都医科大学附属北京儿童医院血液肿瘤中心确诊NB且经荧光原位杂交法确定伴有MYCN扩增的患儿,分析患儿瘤灶部位、大小、转移部位、肿瘤标记物、病理亚型、治疗情况及影响预后的相关因素.主要结局指标3年生存影响因素.结果 纳入133例MYCN扩增的NB患儿,占同期收治总NB患儿的12.0％.男82例,女51例,中位发病年龄(35.7±9.8)个月;原发瘤灶位于腹膜后及肾上腺区129例(97.0％),位于后纵隔区域4例(3.0％);骨髓转移81例(60.9％),骨骼转移80例(60.2％),中枢转移24例(18.1％);99例(74.4％)血清LDH≥1500 U·L-1,126例(94.7％)神经元特异性烯醇化酶≥100 ng·mL-1;原发瘤灶最大直径≥10 cm者89例(66.9％).3年OS和EFS分别为(19.7±3.5)％和(19.0±3.6)％.78例进展复发,在诱导、巩固、维持及停药后进展复发分别为8、20、46和4例,进展复发的部位以原发瘤灶、骨髓、中枢神经系统及骨骼最常见,中位首次进展时间为11.3月.伴有骨髓、骨转移、合并1p36缺失、年龄<18月及未行自体外周血造血干细胞移植患儿预后不良.结论 伴有MYCN扩增的NB患儿原发瘤灶以腹膜后肾上腺区为主,早期远处转移率高,50％以上患儿在维持治疗期间肿瘤进展,3年OS仅为19.7％.伴有MYCN扩增的NB患儿迫切需要靶向治疗等新的治疗手段,以提高疗效,改善预后.

Objective:To explore the clinical significance of the MYCN gene, PHOX2B gene and plasma cell-free DNA (cfDNA) in risk stratification and predicting the prognosis of high-risk neuroblastoma (NB). Methods:This was a prospective study involving 94 high-risk NB children admitted to Beijing Children′s Hospital, Capital Medical University from August 2017 to December 2018.Relative levels of MYCN and PHOX2B and cfDNA at diagnosis, and 4 and 6 cycles of chemotherapy were detected, and their differences were compared by the Chi- square test.Kaplan-Meier survival analysis was performed to explore their prognostic potential in high-risk NB. Results:Among the 94 high-risk NB children, 14 cases (14.9%) had MYCN amplification, 76 cases (80.8%) had positive expression of PHOX2B and 56 cases (59.6%) had cfDNA level higher than 100 μg/L.The proportion of high lactate dehydrogenase (LDH, ≥1 500 U/L) level in the MYCN gene amplification group (6/14 cases) was higher than that in the normal group (9/80 cases) ( P=0.009). The proportion of multi-site metastasis (54/76 cases) and high neuron specific enolase (NSE) level (NSE≥370 μg/L, 37/76 cases) in PHOX2B positive group were significantly higher than those in the negative group (5/14 cases, 2/14 cases) ( P=0.015, 0.020). The proportion of high LDH and high NSE in high cfDNA concentration (≥229.6 μg/L)group (13/37 cases, 28/37 cases) were significantly higher than those in low cfDNA concentration group (2/48 cases, 10/48 cases) (all P<0.001). With the decreased tumor burden during the treatment, the copy number of PHOX2B gene and cfDNA level were significantly lower than those at the initial diagnosis [0 (0-719.6) copies vs.1 723.5 (0-186 000.0) copies; 19.0 (1.1-225.5) μg/L vs.200.6 (8.0-5 247.4) μg/L, all P<0.001]. The 2-year event-free survival (EFS) rate of the MYCN gene amplification group was significantly lower than that of the normal group[(33.3±13.1)% vs.(58.5±7.1)%, P=0.020]. The 2-year EFS rate of PHOX2B positive group was significantly lower than that of the negative group[(47.9±7.1)% vs.(79.1±11.1)%, P=0.043]. EFS rate in high cfDNA concentration group was significantly lower than that in cfDNA low concentration group[(38.6±9.8)% vs.( 71.7±8.2)%, P=0.001]. After 6 cycles of chemotherapy, EFS rate in the PHOX2B positive group was significantly lower than that in the negative group [(16.7±14.4)% vs.( 60.6±6.6)%, P=0.014]; which was significantly lower in the Metaiodobenzylguanidine (MIBG) positive group than that of the negative group[(35.2±11.7)% vs.(65.8±7.1)%, P=0.037]. The MYCN gene and cfDNA concentration were not correlated with the prognosis of high-risk NB.Survival analysis of the combination of PHOX2B and MYCN gene ( PHOX2B+ /MIBG + , PHOX2B+ or MIBG + , PHOX2B-/MIBG -) showed a significant difference in the survival among three groups[0 vs.(53.6±1.2)% vs.(65.5±7.4)%, P=0.003]. Conclusions:The MYCN and PHOX2B gene and cfDNA concentration are of significance in risk stratification and predicting the prognosis of high-risk NB.Compared with the MYCN gene and cfDNA concentration, the PHOX2B gene is more suitable for monitoring the curative effect of chemotherapy on high-risk NB.A combined analysis of PHOX2B gene and MIBG before treatment can be more accurate in evaluating the treatment effect and residual lesions.

目的 定位C57BL/6小鼠胫骨前肌(TA)和趾长伸肌(EDL)肌梭分布,分析肌梭在骨骼肌中的固定方式,并统计肌梭各区域长度和赤道横截面积(CAS)等参数分析肌梭形态学共性,为肌梭的形态和功能研究提供解剖学基础.方法 5只正常成年C57BL/6小鼠取TA和EDL,利用改良的骨骼肌冷冻技术得到无冰晶样本,样本连续冷冻切片,HE染色,显微成像,分析TA与EDL肌梭分布及肌梭在骨骼肌中与其他组织的连接方式.测量肌梭各区域长度和CAS,统计学分析肌梭形态特征.结果 C57BL/6小鼠TA和EDL分布情况为:从尾端至头端方向,肌梭主要分布在肌腹中间偏上位置.从背侧至腹侧,肌梭靠近腓深神经入肌点分布.在肌梭末梢可以看到肌梭锚定连接梭外纤维并固定于骨骼肌中.单个肌梭形态观察分析显示,连接感觉神经纤维末梢的区域A和与运动神经纤维末梢连接区域B的长度有较明显相关性(相关系数为0.75).结论 C57BL/6小鼠TA和EDL肌梭分布特点可以为后续肌梭相关的形态学和电生理研究提供解剖信息,发现肌梭区域A和区域B的相关性可能有助于解释肌梭信号传递能力差异.

目的 探索病理免疫组化方法检测MYCN蛋白的表达与荧光原位杂交法方法检测MYCN基因的一致性,初步建立价格低廉、操作简单且易于推广的MYCN检测手段,用于神经母细胞瘤患儿早期的分层诊疗.方法 应用FISH及IHC共同检测NB患儿的肿瘤组织及转移骨髓的MYCN表达.结果 共54例NB患儿纳入本研究,全部患儿的瘤组织均经FISH方法检测,其中MYCN基因扩增31例,非扩增23例;入组患儿中有25例患儿病初伴有骨髓转移,对转移骨髓应用FISH及IHC方法检测,17例患儿MYCN蛋白表达阳性,8例为阴性,与患儿瘤组织FISH方法检测结果完全一致;用IHC方法检测肿瘤组织及转移骨髓的MYCN蛋白,阳性一致率均为86％～94％,阴性一致率为100％.结论 IHC方法对NB患儿的肿瘤组织及受累转移的骨髓进行MYCN蛋白的检测,其结果与FISH方法的检测结果具有较高的一致性,价格低廉、操作简单,适用于NB患儿早期的危险度评估及推广.

背景:大鼠光化学栓塞模型可以良好地模拟缺血性卒中,具有损伤脑区特异性、高重复性及低死亡率等优势,但光源的选择及玫瑰红注射浓度等多个条件都会影响造模的结果,需要后期进行细致的评估来确定造模的成功.目的:建立稳定、不易自发恢复的大鼠缺血性脑卒中模型,并探究卒中区域的病理变化及大鼠行为学变化.方法:雄性Wistar大鼠52只随机被分为假手术组6只、光化学栓塞组46只.光化学栓塞组取18只分别于大鼠股静脉注射20,40及80 mg/kg玫瑰红制作光化学栓塞模型,筛选出造模最佳的玫瑰红浓度;再对剩余28只大鼠进行光化学栓塞造模手术;假手术组大鼠不进行激光器定点照射及不注射玫瑰红染料.术后1d利用TTC染色揭示在不同玫瑰红浓度注射下梗死范围的变化;对大鼠光化学栓塞后1,3,7,14 d运用NeuN染色观察梗死区域内神经元的死亡情况;Iba-1,GFAP,GLUT-1染色观察光化学栓塞后卒中区域炎症反应、胶质瘢痕及血管的变化;应用圆柱体实验及网格错误实验评价大鼠光化学栓塞后行为学的变化.结果 与结论:①80 mg/kg的玫瑰红浓度引起的卒中腔体积最大,光化学栓塞模型重复性高,动物死亡率低,7d可以形成较稳定的卒中腔;②光化学栓塞后在7d可以形成相对稳定的胶质瘢痕带,但炎症反应在1-14 d逐渐加重;③光化学栓塞减少卒中腔附近的血管面积,并在14 d趋于稳定;④光化学栓塞后大鼠出现长期的感觉及运动功能下降;⑤结果表明大鼠光化学栓塞模型稳定良好,不易自发恢复,适合缺血性脑卒中病理变化的研究.

背景 视网膜母细胞瘤(Rb)是儿童期最常见的恶性肿瘤,在90％Rb患儿所在的发展中国家,减少死亡仍是一个挑战,且患儿的生存质量较少受到关注.目的 总结单眼眼内期Rb患儿疗效、安全性及生存质量,为优化治疗方案和提高治疗安全性及有效性提供证据.设计回顾性非随机对照研究.方法 回顾性分析2009年11月6日至2019年9月5日首都医科大学附属北京儿童医院收治的单眼眼内期Rb患儿的临床资料,末次随访日期为2020年4月20日.化疗方案为根据加拿大多中心RB 2003方案改良的北京儿童医院RB-2009方案,并应用810 nm及532 nm激光、冷凝器等技术进行局部治疗.分析保眼率、生存率、死亡原因、不良反应等,并对随访时≥5岁儿童采用儿童生存质量普适性核心量表和儿童生存质量癌症模块量表进行生存质量调查.主要结局指标5年总生存率(OS)和生存质量.结果 符合纳入标准的6262例Rb,完成随访168例,中位随访时间51(8～125)月,平均化疗疗程4.19±2.16(1～12)疗程.B期1例,C期3例,D期95例,E期69例.共眼球摘除89只(53.0％),其中D期42例,E期47例;直接眼球摘除30例(33.7％),化疗后眼球摘除59例(66.3％).成功保眼79例(47.0％),其中D期53例(55.79％),E期22例(31.9％).预计全部患儿5年总生存率(OS)为95.2％.眼球摘除和保眼治疗患儿5年OS分别为95.5％和94.9％,差异无统计学意义(P=0.78).5例(3.0％)发生可逆性Ⅰ级听力损害,无Ⅱ～Ⅳ级听力损害发生;所有患儿均发生骨髓抑制,6例次(3.6％)输注PLT,无感染相关死亡发生.106例≥5岁患儿完成生存质量调查,眼球摘除患儿对外貌的自我感觉维度、社会维度和角色维度平均得分均低于保眼患儿,差异均有统计学意义(P<0.05).死亡8例,D期5例,E期3例,2例因经济原因放弃治疗后死亡,6例复发后颅内转移死亡.结论 VEC(长春新碱、依托泊苷和卡铂)方案全身化疗结合局部治疗对单眼眼内期Rb患儿是安全、有效的.颅内转移仍然是Rb相关死亡最常见的原因.≥5岁眼球摘除患儿在外貌自我感觉、社会、角色维度生存质量下降.

背景:前期研究显示,神经营养因子3（neurotrophin 3,NT3）-壳聚糖可诱发脊髓损伤大鼠内源性神经发生和轴突再生,促进大鼠运动和感觉功能恢复。目的:观察康复训练结合NT3-壳聚糖活性生物材料支架对完全性脊髓损伤大鼠骨骼肌形态变化和功能恢复的影响。方法:将50只成年雌性Wistar大鼠随机分为5组,每组10只:假手术组不造模,其余4组制备T7-T8全切5 mm脊髓损伤模型,单损组造模后不进行任何干预,另3组分别给予康复训练、NT3-壳聚糖活性生物材料支架、NT3-壳聚糖活性生物材料支架结合康复训练干预,康复训练于造模后2周开始。造模前及造模后2,4,6,8,10,12周对各组大鼠进行开放场地的BBB评分;造模后12周,取后肢骨骼肌（胫骨前肌、腓肠肌、比目鱼肌）进行苏木精-伊红和乙酰胆碱酯酶染色,评定各组大鼠肌肉萎缩和运动终板的变化情况。实验方案经首都医科大学动物实验委员会批准（批准号为AEEI-2018-105）。结果与结论:①假手术组术后各时间点的BBB评分高于其他4组（P ＜0.05）,NT3-壳聚糖结合康复训练组造模后8,10,12周的评分高于单损组、单损结合康复训练组、NT3-壳聚糖组（P ＜0.05）;②造模后12周苏木精-伊红染色显示,造模4组的各骨骼肌肌纤维横截面积和直径小于假手术组（P ＜0.05）,其中NT3-壳聚糖结合康复训练组各骨骼肌肌纤维横截面积和直径大于单损组、单损结合康复训练组、NT3-壳聚糖组（P ＜0.05）;③造模后12周乙酰胆碱酯酶染色显示,造模4组各骨骼肌的运动终板乙酰胆碱酯酶平均吸光度值均低于假手术组（P ＜0.05）,其中NT3-壳聚糖结合康复训练组高于单损组、单损结合康复训练组、NT3-壳聚糖组（P ＜0.05）;④结果表明,康复训练结合NT3-壳聚糖活性生物材料支架植入能有效防止完全性脊髓损伤大鼠后肢骨骼肌肌肉萎缩,提高运动终板乙酰胆碱酯酶活性,减轻神经肌肉接头退变,改善大鼠后肢运动功能。

Objective:To summarize the causes of death and severe complication in the early diagnosis of children with neuroblastoma (NB), and to analyze the relative factors of early death of children with NB, so as to raise awareness and reduce early mortality by early detection and early intervention.Methods:Patients with newly diagnosed NB in the Hematology Oncology Center of Beijing Children′s Hospital from April 2007 to December 2017 were included consecutively, and those died within 1 month after diagnosis were retrospectively analyzed.The general data of patients, immediate causes of death, complications, time elapsed between death and diagnosis, whether to receive chemotherapy and other information were collected.Results:A total of 654 cases were included for diagnosis, treatment and follow-up, 31 cases of which died in early stage, accounting for 4.7% of the total.The major complication were pulmonary infection in 18 cases (58.1%) and bone marrow suppression after chemotherapy in 17 cases (54.8%), tumor rupture hemorrhage in 16 cases (51.6%), multiple organ failure in 8 cases (25.8%). Risk factor analysis of the 31 early death cases with NB was conducted.Single factor analysis: there were statistical differences between early death group and non-early death group in risk grouping ( P=0.006 6), bone marrow invasion ( P=0.020 7), site of primary tumor ( P=0.016 7), age ( P=0.003 3), lactate dehydrogenase (LDH) level ( P<0.000 1), neuron-specific enolase (NSE) level ( P<0.000 1), serum ferritin level ( P=0.016 0), D dimer level ( P<0.000 1), fibrinogen level ( P=0.002 7), diameter of tumor ( P<0.000 1), hemoglobin ( P<0.000 1), platelet level ( P<0.000 1), serum albumin level ( P<0.000 1). Multiple-factor analysis: age younger than 30 months, OR=2.824 (95% CI: 1.084-7.359), LDH level greater than 1 004 IU/L, OR=6.991 (95% CI: 2.135-22.887), albumin level less than 36 g/L, OR= 65.237 (95% CI: 2.024-13.545), hemoglobin level less than 92 g/L, OR=5.358 (95% CI: 2.024-13.545), platelet level less than 192×10 9/L, OR=3.554 (95% CI: 1.267-9.965). Conclusions:Strengthening vital signs detection after admission, identifying severe life-threatening complications such as rupture of tumors as early as possible, implementing symptomatic interventions such as appropriate sedation and active transfusion of blood products as early as possible after invasive operation, and transferring to intensive care unit for respiratory support when necessary are important means to avoid early death.

目的:分析顺行与逆行交叉克氏针髓内固定治疗不稳定的第五掌骨颈骨折的临床效果.方法:采用回顾性分析方法,按照不同的手术方式将本院2017年1月至2019年9月收治的30例不稳定性第五掌骨颈骨折患者分为A组(n=13)和B组(n=17),A组接受顺行克氏针髓内固定治疗,B组接受逆行交叉克氏针髓内固定治疗.比较两组手术复位效果、术后并发症、术后的头干角、健侧头干角、患侧及健侧掌指关节主动屈曲及背伸活动范围、上肢功能评分(DASH).结果:两组病例术后均恢复良好,未出现明显并发症,在末次随访时所有病例骨折愈合均达到临床愈合.A组解剖复位率7.69％,B组解剖复位率17.65％,差异无统计学意义(P=0.892).术前、术后,两组头干角比较,差异均无统计学意义(P>0.05).术后4个月、术后6个月,两组屈曲活动范围、背伸活动范围均大于本组术前;DASH评分均低于本组术前(P<0.05).术后4个月,A组屈曲活动范围、背伸活动范围大于B组;DASH评分低于对照组(P<0.05).结论:两种术式均可达临床愈合,复位率良好,顺行克氏针髓内固定术后4个月活动范围优于逆行交叉克氏针固定治疗.

背景:大鼠大脑中动脉末端闭塞模型可以引起稳定的局灶性皮质梗死,在很好模拟人类卒中病理状态的同时死亡率较低,但对手术人员的技术以及设备有较高要求,并且需要后期评估才能确定造模成功与否.目的:建立有效稳定简便的缺血性小鼠脑卒中模型;揭示小鼠脑卒中后梗死区及周围区域的病理变化;探索小鼠脑卒中后的行为学改变.方法:对小鼠末端大脑中动脉进行永久电凝结扎,24 h后用TTC染色明确该模型梗死范围并统计该模型的成功概率;对小鼠大脑中动脉末端闭塞模型后不同时间点(1,3,7,10,14 d)的脑组织切片进行苏木精-伊红染色,观察缺血坏死区的体积在不同时间点的改变;进行胶质纤维酸性蛋白、Iba-1免疫组织化学染色,检测模型小鼠脑损伤后胶质反应及炎症反应变化;最后应用网格足部错误试验和圆柱体试验评价小鼠卒中后感觉运动功能的缺失情况.结果与结论:①大脑中动脉末端闭塞模型导致局灶性皮质梗死且该新模型死亡率仅9％,成功率达87％;②梗死区域主要位于M1/S1/S2区,梗死范围在闭塞10 d时趋于稳定;③脑卒中后3 d时炎症反应达到高峰,14 d时损伤区周围形成稳定星形胶质瘢痕;④局灶性皮质梗死后小鼠对侧肢体出现明显的感觉及运动的缺失;⑤结果表明,卒中后实验小鼠立即出现感觉及运动功能缺失,且持续至卒中后12周;实验建立的大脑中动脉末端闭塞模型稳定、可靠,梗死范围明确,适合缺血性脑卒中的研究.

目的 总结神经母细胞瘤(NB)患儿应用铂类药物化疗后毒性反应和药物基因多态性检测结果,探索二者之间的关联性,为指导临床个体化治疗提供依据.方法 连续纳入2016年2月1日-2017年5月31日期间,北京儿童医院血液肿瘤中心确诊并系统治疗的NB患儿.依据BCH-NB-2007危险度分组标准,分为低危、中危、高危组,接受含顺铂或卡铂方案化疗.化疗前留取外周血,采用荧光杂交方法对两种铂类化疗药物基因组DNA进行检测,化疗后记录患儿各系统毒副反应,并按国立癌症研究所常规毒性标准(NCI-CTCAE 5.0中文版)进行分级.进一步将3/4级毒性反应与药物基因检测结果进行关联性分析.结果 共纳入98例NB患儿,男50例,女48例.中位年龄为46(5 ～115)个月.中低危组单用卡铂者36例(36.7％).高危组62例(63.3％),其中单用顺铂者46例,顺铂+卡铂者16例.所有NB患儿在病初时留取血标本检测GSTP1基因多态性,AA型61例(62.2％),AG型33例(33.7％),GG型4例(4.1％);部分患儿(51例)还进行了XPC基因多态性的检测,其中GG型8例(15.7％),GT型24例(47.1％),TT型19例(37.3％).所有NB患儿中,出现3/4级毒性反应共61例(62.2％),其中以3/4级血液毒性最为常见(55例,56.1％);3/4级恶心呕吐、肝损害及电解质紊乱各2例,共占6％.分析3/4级毒副反应与所检测的两种药物基因多态性之间的相关性,GSTP1基因多态性与3/4级毒性反应相关,组间差异具有显著性,AA型毒副反应重于AG型和GG型(P＜0.05);XPC基因多态性与3/4级毒性反应相关性差异无显著性.结论 NB患儿应用铂类化疗毒性反应以血液系统毒性、消化道毒性为主,GSTP1基因多态性与3/4级毒性反应相关,AA型毒副反应最重;XPC基因多态性与3/4级毒性反应无相关性,有待扩大样本量加以验证.

背景与目的 骨髓细胞的双色间期荧光原位杂交(interphase fluorescence in situ hybridization,FISH)被证实是研究骨髓转移性神经母细胞瘤患者v-myc禽类骨髓细胞瘤病毒癌基因神经母细胞瘤衍生同源基因(v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog,MYCN)扩增的直接、有效的方法.然而,单凭MYCN扩增不足以进行预处理风险分层.最近,染色体11q23缺失被纳入神经母细胞瘤的风险分层中.在本研究中,我们旨在研究11q23缺失和MYCN扩增在骨髓转移性神经母细胞瘤患者的生物学特性及其对预后影响.方法 我们利用骨髓细胞的双色间期FISH方法分析了101例骨髓转移性神经母细胞瘤患者的MYCN和11q23状态,并比较了两种畸变的生物学特征和预后影响.结果 有12例(11.9％)和40例(39.6％)患者分别出现MYCN扩增和11q23缺失.这两个标志物几乎不同时出现.MYCN扩增主要发生在乳酸脱氢酶(lactate dehydrogenase,LDH)和神经元特异性烯醇化酶(neuron-specific enolase,NSE)水平升高的患者中(P<0.001);与MYCN-正常患者相比,MYCN扩增的患者多伴有事件发生(如肿瘤复发、进展或死亡)(P=0.004).11q23缺失仅与年龄相关(P=0.001).与MYCN正常患者相比,MYCN扩增患者的预后较差[3年无事件生存率(event-free survival,EFS):8.3±8.0％vs.43.8±8.5％,P<0.001;3年总生存率(overall survival,OS):10.4±9.7％vs.63.5％ ±5.7％,P<0.001).11q23缺失仅在MYCN正常患者预后不良(3年EFS率:34.3±9.5％v s.53.4±10.3％,P=0.037;3年OS率:42.9±10.4％vs.75.9±6.1％,P=0.048).同时具有MYCN扩增和11q23缺失的患者预后最差(P<0.001).结论 染色体11q23缺失仅在无MYCN基因扩增的骨髓转移性神经母细胞瘤预示预后不良.在识别高危患者方面,两种标志物的联合评估远优于单一标志物评估.

Objective:To summarize and analyze the results of chromosome karyotype in children with neuroblastoma (NB) with bone marrow metastasis at first diagnosis, and to discuss the clinical significance.Methods:G-banding was applied to the analysis of chromosome karyotype of patients who were regularly treated in the Hematological and Oncology Center in Beijing Children′s Hospital from January 2015 to December 2017, and all the patients were followed up until December 31, 2018.Their clinical features and prognosis were analyzed.Results:(1) There were 120 cases with bone marrow metastasis, including 74 boys and 46 girls, and 98 cases (81.7%) were ≥ 18 months.Among 60 cases with normal chromosome, 56 cases (93.3%) were in International Neuroblastoma Staging System(INSS)-Ⅳ phase, and 4 cases in INSS-Ⅳs phase; there were 2 low-risk (LR) cases, 9 intermediate-risk (MR) cases, and 49 high-risk (HR) cases (81.7%); 7 cases had MYCN gene amplifications.All 60 patients with chromosome abnormalities were in INSS-Ⅳ phase; there was 1 case in MR and 59 cases (98.3%) in HR; 14 cases had MYCN gene amplifications.(2) Among 60 children (50%) with chromosome abnormalities, 4 children had number abnormalities, 14 children had structural abnormalities, and 42 children had both number and structural chromosome abnormalities.Chromosome 21, 10, 11 deletions were the most common in number abnormalities; structural abnormalities involving 11q, 1p, 3p segments had a high incidence.(3) Seventeen cases of children with normal chromosome had tumor progression or recurrence during the 4 to 44-month follow-up period, and 31 cases of children with chromosome abnormalities had tumor progression or recurrence during the 2 to 42-month follow-up period.The 3-year overall survival rate and event-free survival rate of all children were 60.0% and 48.4%, respectively; children in the normal chromosome group had a 3-year overall survival rate of 74.2% and an event-free survival rate of 65.7%; the 3-year overall survival rate and event-free survival rate of children with chromosome abnormalities were 47.5% and 24.9%, respectively.Most children suffering from tumor progression or recurrence had chromosome 10 deletion, and abnormal structure of 11q, 1p, 2p segments. Conclusion:The chromosomal abnormality rate of Nb children's tumor cells is high, but the repetition rate is low, and the individual difference is obvious.The deletion of chromosome 10, abnormal regional structure of 11q, 1p and 2p segments may be poor prognostic factors for NB.Chromosome karyotype analysis of bone marrow samples is feasible, which can provide a basis for more accurate risk stratification and treatment.

目的:通过伪狂犬病毒(PRV)逆行示踪技术观察Wistar大鼠两侧背根神经节(DRG)之间神经元的联系,寻找两侧DRG神经元交互支配的形态学基础.方法:选用18只正常成年雌性Wistar大鼠,分为单侧PRV组、双侧PRV组、霍乱毒素B亚单位(CTB)组(n=6).单侧PRV组将2μl滴度为1×1010的PRV-EGFP注射到左侧坐骨神经;双侧PRV组将2 μl滴度为1×1010的PRV-EGFP注射到左侧坐骨神经上,同时2μl滴度为1×1010的PRV-mRuby注射到右侧坐骨神经上;CTB组将2μl浓度为1μg/μl的CTB注射到左侧坐骨神经上.5d后观察病毒标记结果.结果:单侧PRV组在左侧DRG可见大量被PRV-EGFP标记的神经元,右侧DRG也观察到大量被PRV-EGFP标记的神经元,但数量明显少于左侧(P＜0.01).双侧PRV组在左侧DRG可见大量被PRV-EGFP和PRV-mRuby标记的神经元,且有部分神经元被PRV-EGFP和PRV-mRuby同时标记.CTB组大鼠L4脊髓前角神经元被CTB标记,DRG中枢突大量纤维终末被CTB标记.此外,L2脊髓中背根神经节中枢突的大量纤维终末被CTB标记.单侧PRV组大鼠L4脊髓前角神经元被PRV-EGFP标记,腰骶髓后连合核(DCN)有大量神经元被PRV-EGFP标记,L2腰骶髓后连合核也有大量神经元被PRV-EGFP标记.结论:Wistar大鼠的左右侧DRG神经元之间存在交互支配的现象,两侧DRG神经元之间并没有直接的突触联系,而是通过后连合核等中间神经元跨突触联系.

目的 观察评估脊髓损伤后恒河猴的下肢残留跨步能力.方法 成年雌性恒河猴4只,胸椎T7-9右半侧切除脊髓组织1 cm.分别在脊髓损伤前、脊髓损伤后6周和12周,采用VICON系统进行双下肢步态测试,获取动物在跑步机上连续跨步中的双下肢步态周期时长,步长、步高、膝/踝关节角度幅值以及联动参数比值,并量化分析.结果 脊髓损伤后,恒河猴双下肢的协调性破坏,右下肢明显拖拽;左下肢步态周期时长显著增加(P<0.001),膝/踝关节角度屈曲/伸展的幅值均显著增大(P<0.001).联动参数比值在脊髓损伤前后均无显著性差异(P>0.05).左下肢步态周期时长与步长(r=0.838,P=0.001)、步高(r=0.726,P=0.007)和踝关节角度变化幅值(r=0.766,P=0.004)均相关,踝关节角度变化幅值与步长呈正相关(r=0.627,P=0.029).结论 脊髓损伤后恒河猴健侧肢体步态模式发生改变.健侧下肢代偿性调整运动策略,以适应患侧下肢功能的缺失.