小儿肝脏恶性肿瘤血管再生性特点的研究

目的：研究小儿原发性肝脏恶性肿瘤中血管内皮生长因子（VEGF）和微血管密度（MVD）的特点。方法：采用SPTM免疫组化方法检查14例小儿原发性肝脏恶性肿瘤VEGF阳性表达以及MVD计数。病理分类：肝细胞癌7例，肝母细胞瘤5例，恶性间叶瘤和横纹肌肉瘤各1例。另选12例成年人肝癌手术切除标本作为对照。结果：小儿肝肿瘤组VEGF阳性表达面积显著高于成人肝癌组（P＜0．05）；尤其小儿肝癌组与成人组对比差异有极显著性意义（P＜0．01）。小儿肝肿瘤组的MVD也显著高于成人肝癌（P＜0．05）。小儿肝癌组患儿均在2年内死亡；非肝癌肝脏肿瘤预后较好，2例存活超过5年。结论：小儿肝脏恶性肿瘤存在有高度血管再生性的特点，肿瘤的生长快，预后差，以小儿肝癌最为突出。长期存活患儿的VEGF表达均为阴性。     

小儿肝脏血管内皮细胞瘤组织中VEGF表达的研究

目的：研究血管内皮生长因子（VEGF）、CD34在小儿肝脏血管内皮细胞瘤（IHHE）组织中的表达，探讨VEGF在血管内皮细胞瘤诊断和治疗中的作用。方法14例肝脏血管内皮细胞瘤石蜡标本来自2002年7月至2009年7月北京儿童医院手术切除并经病理诊断的临床病例，术前未行其他治疗，并选取小儿肝脏恶性肿瘤组织（肝母细胞瘤20例），其他肝脏良性肿瘤组织（结节样增生10例，错构瘤10例）及IHHE瘤旁组织作为对照组；空白对照选取良性肿瘤术中切取部分肝脏组织。组织切片SP法进行免疫组织化学染色，兔抗VEGF和CD34相关抗原标记；计算出VEGF表达阳性率，CD34相关抗原抗体检测出肿瘤间质血管，分析其表达的不同及相互关系。应用统计软件进行数据分析。结果①VEGF在肝母细胞瘤组织中表达率为90.0％（18/20），在良性肿瘤组中表达阳性率为10.0％（2/20）；VEGF在正常肝组织、瘤旁组织中呈低水平、稳定表达，染色浅，而在IHHE肿瘤组织中则呈阴性表达。在肝母细胞瘤组阳性表达率明显高于其他组，差异有显著性（P＜0.05），其余两两比较，差异均无显著性（P＞0.05）。②CD34在已知正常肝组织肝窦处未见阳性表达；在IHHE组织中的阳性表达率为100.0％（14/14），肝母细胞瘤为90.0％（18/20），肝脏良性肿瘤为15.0％（3/20），瘤旁组织未见表达。MVD在IHE，肝母细胞瘤组，良性肿瘤组及瘤旁组织分别为31.55±4.86，29.75±5.56，11.23±3.97。结论 IHHE中，高表达CD34，低表达VEGF，表明其不是单纯的血管、内皮细胞病变，很有可能是血管瘤与血管畸形的混合体。IHE增殖期抗血管生成治疗可能促进肿瘤退化。     

化疗对小儿神经母细胞瘤的VEGF表达的影响

目的研究化疗对小儿神经母细胞瘤的VEGF表达以及血管形成状态的影响。方法收集自1993年12月~2000年12月住院治疗的神经母细胞瘤患儿中保存完好的60例肿瘤组织的石蜡标本(包括手术切除和肿瘤活检),按术前有无化疗情况分为术前化疗组(34例)和未化疗组(26例),进行VEGF和CD31免疫组化分析。结果60例标本中VEGF阳性表达36例,其中术前未化疗组VEGF表达阳性率为76.9%(20/26);术前化疗组VEGF表达阳性率为47.1%(16/34)。①两组VEGF表达阳性率比较χ2=5.84,P<0.05。②有远处转移组(Ⅳ期)与无远处转移组(Ⅰ、Ⅱ、Ⅲ期)VEGF表达阳性率比较2χ=4.051,P<0.05。③微血管计数未化疗组为25.73±5.85,术前化疗组为15.64±5.17,t=7.08,P<0.001。结论神经母细胞瘤转移与肿瘤组织中VEGF过度表达有密切关系。术前化疗能抑制神经母细胞瘤组织中的VEGF表达,从而抑制肿瘤新生血管形成,防止肿瘤转移和复发。     

双酚A对神经母细胞瘤SK-N-SH细胞株移植瘤生长的影响

目的环境内分泌干扰物对人类健康的影响已引起全球的关注。本研究旨在观察双酚A（BPA）对神经母细胞瘤生长的影响及其可能机制。方法常规培养SK-N—SH细胞，接种于双侧卵巢切除（OVX）的裸小鼠皮下。可触摸到肿瘤结节时，将裸鼠随机分为3组：①对照组（OVX＋溶剂，OVX，n=9）；②E2组（OVX＋E2，500ug·kg^-1·d^-1，n=11）；③BPA组（OVX＋BPA，200mg·kg^-1·d^-1，BPAgroup，n=10）。每周2次检测皮下移植瘤体积（TV）和裸鼠体重。第18d，解剖处死所有裸鼠，称移植瘤重量。免疫组化方法检测肿瘤组织微血管内皮细胞的表达情况，在光学显微镜高倍视野下分别计数肿瘤组织的微血管密度（MVD））。用Western Blot方法检测肿瘤组织血管内皮细胞生长因子（VEGF）蛋白的表达情况。结果实验终点时，OVX组、E2组和BPA组的TV分别为（2．62±0．54）cm^3、（5．07±0．93）cm^3和（4．19±0．68）cm^3，BPA组较OVX组增高59％（P〈0．05）。3组裸鼠体重没有明显差异（P〉0．05）。3组瘤重分别为（1．64±0．18）g、（2．58±0．19）g、（2．96±0．27）g，BPA组较OVX组增高82％（P〈0．05）；3组中Ⅷ因子相关抗原均染色，E2组（22．73±1．46）及BPA组（27．76±2．24）MVD较对照组（15．14±1．12）明显增高（P〈0．01），VEGF的蛋白表达亦较对照组增高（P〈0．05）。结论BPA可促进人神经母细胞瘤裸鼠皮下移植瘤生长，微血管形成可能参与了促生长作用。     

化疗及非化疗组神经母细胞瘤干细胞标志物的表达及意义

目的探讨干细胞标志物CD133、ABCG2、CD117及nestin在神经母细胞瘤化疗组及非化疗组患儿中的表达及意义。方法收集上海交通大学附属新华医院小儿外科保存完好的20例神经母细胞瘤肿瘤组织的石蜡标本,按术前是否经历化疗分为术前化疗组和未化疗组,每组各10例,采用免疫组化SupperVision法检测4种标记物在肿瘤组织中的表达。结果CD133、ABCG2、CD117及nestin干细胞标志物在未化疗神经母细胞瘤组织中的平均阳性细胞率低于化疗后残留神经母细胞瘤组织中阳性细胞率的平均值,差异无统计学意义（P〉0．05）。两组患儿年龄、性别、分期及组织分型比较,差异无统计学意义（P〉0．05）。结论儿童神经母细胞瘤中可能存在肿瘤干细胞,肿瘤干细胞对常规化疗药物具有耐药性。     

采用交替或多药联合化疗治疗晚期神经母细胞瘤

目的：观察交替化疗与多药联合化疗治疗晚期儿童神经母细胞瘤的疗效,毒性反应及对生存期的影响。方法：对29例晚期神经母细胞瘤的患儿给予术前或术后化疗,14例采用COAVIP或VP方案交替化疗共155疗程,15例采用COPE等多药联合化疗共84疗程。结果：交替化疗组14例可评价疗效13例,CR0,PR12例,NC1例,PD0。有效率12／13（92．3％）。化疗后手术完整切除率10／13（76．9％）。多药联合化疗组15例,可评价疗效11例,CR1例,PR5例,NC3例,PD2例,有效率6／11（54．5％）。化疗后手术切除率为5／11（45．5％）。两组比较P＜0．05（X^2检验）,化疗的毒性反应主要表现为骨髓抑制,恶心呕吐等,均为可逆尾。两组生存期比较经秩和检验P＞0．05无统计学差异。结论：交替化疗较之多药联合化疗治疗晚期儿童神经母细胞瘤疗效好,手术切除率高。毒性反应大致相同,两组生存期也大致相同。     

神经母细胞瘤脂类提取物对NKT细胞激活作用的实验研究

目的 研究神经母细胞瘤脂类提取物对健康儿童自然杀伤T（NKT）细胞增殖程度和对NKT细胞介导的对自身肿瘤细胞的细胞毒作用的影响。方法 甲醇／氯仿法提取神经母细胞瘤脂类提取物作为刺激物，采集健康儿童外周血单核细胞，培养出成熟的树突细胞，采集并分离同一个体外周血T细胞与树突细胞共同培养，分别加入脂类提取物或（和）白细胞介素15（IL－15）和粒－单系细胞集落刺激因子（GM－CSF）。测定各组T细胞的增殖程度的变化，流式细胞仪测定NKT细胞占T细胞的比例（NKT／T）并分选出NKT细胞对神经母细胞瘤细胞毒作用。结果 肿瘤脂类提取物刺激组T细胞增殖程度、NKT／T比例、NKT细胞毒作用高于对照组（P＝0．043；P＝0．020；P＝0．032）。共刺激组（肿瘤脂类提取物＋IL－15＋GM－CSF）T细胞增殖程度、NKT／T比例、NKT细胞毒作用高于对照组（P＝0．001；P＝0．001；P＝0．020），肿瘤脂类提取物刺激组（P＝0．004；P＝0．030；P＝0．010）和细胞因子刺激组（P＝0．044；P＝0．049；P＝0．048）。结论 神经母细胞瘤脂类提取物可以有效激活NKT细胞，发挥对自身肿瘤细胞的细胞毒作用。细胞因子IL－15和GM－CSF可以增强脂类提取物的激活作用。     

术前化疗对神经母细胞瘤细胞凋亡和增殖的影响作用

目的：探讨术前化疗对神经母细胞瘤细胞凋亡和肿瘤增殖的作用。方法：应用脱氧核糖核酸末端转移酶标记（ＴＵＮＥＬ）和增殖细胞核抗原（ＰＣＮＡ）免疫组化，观察术前化疗组６例和一期手术组１１例神经母细胞瘤标本的凋亡指数、增殖指数和分裂指数的差异。结果：术前化疗组凋亡指数（９．１７±４．８８）与一期手术组（１０．１１±５．１６）无显著差异，Ｐ＞０．０５；术前化疗组增殖指数（１８．００±９．５５）明显低于一期手术组（２９．００±１３．８０），术前化疗组的分裂指数（１．６７±１．３４）亦较一期手术组（３．６７±２．２４）有明显下降，Ｐ值均＜０．０５。结论：术前化疗对神经母细胞瘤的肿瘤细胞增殖有明显抑制作用。ＴＵＮＥＬ和ＰＣＮＡ检测为观察肿瘤细胞凋亡和细胞增殖的有效方法     

反义寡核苷酸抑制神经母细胞瘤LA-N-5细胞血管内皮生长因子mRNA表达及其生物效应的研究

摘要 目的 研究血管内皮生长因子（VEGF）反义寡核苷酸（ASODN）转染对神经母细胞瘤LA-N-5细胞VEGF mRNA表达的影响以及对肿瘤细胞增殖、分化的影响。方法 用LipofectamineTM2000介导的VEGF ASODN和错义寡核苷酸（MSODN）转染LA-N-5细胞，半定量RT-PCR检测各组细胞VEGF165和VEGF121 mRNA转染前后不同时间表达的变化；MTT法测定转染后各组细胞的生长曲线及抑制率。结果 半定量RT-PCR检测结果显示，在转染后72 h VEGF165和VEGF121 mRNA的表达：ASODN组为0.346±0.029和0.227±0.036，ASODN+LipofectamineTM2000组为0.275±0.035和0.165±0.017。ASODN组和ASODN+LipofectamineTM2000组均显著抑制VEGF mRNA的表达，ASODN+LipofectamineTM2000组抑制作用较ASODN组更强（P<0.05）；转染后ASODN组和ASODN+LipofectamineTM2000组细胞增殖显著受抑，在48 h时抑制率最高，分别为（39.92±2.74）%及（55.80±2.52）%,ASODN+LipofectamineTM2000组对细胞增殖的抑制作用较ASODN组更为明显（P<0.05）。MSODN组、MSODN+LipofectamineTM2000组对LA-N-5细胞增殖抑制作用与对照组差异无统计学意义（P>0.05）。结论 VEGF ASODN可抑制神经母细胞瘤LA-N-5细胞VEGF在mRNA水平的表达，并能抑制神经母细胞瘤LA N 5细胞的增殖和分化，LipofectamineTM2000能明显增强VEGF ASODN的抑制效果。     

凋亡抑制因子及血管内皮生长因子在儿童肾母细胞瘤中的表达及意义

目的研究凋亡抑制因子（survivin）及血管内皮生长因子（VEGF）在肾母细胞瘤中的表达及其临床意义。方法应用免疫组织化学SABC法及RT-PCR方法研究Survivin及VEGF在52例肾母细胞瘤组织，47例瘤旁组织及8例肾组织在蛋白水平及mRNA水平的表达情况。结果52例肾母细胞瘤组织中Survivin免疫组织化学显示阳性表达率为61．5％（32／52），RT-PCR阳性率为67．3％（35／52）；47例瘤旁组织中免疫组织化学显示阳性表达率为4．3％（2／47），RT-PCR阳性率为6．4％（3／47）；肾组织中未见表达。52例肾母细胞瘤组织中VEGF免疫组织化学显示阳性表达率为75．0％（39／52），RT-PCR阳性率为71．2％（37／52）；47例瘤旁组织中免疫组织化学显示阳性表达率为19．1％（9／47），RT-PCR阳性率为23．4％（11／47）；肾组织中免疫组织化学显示阳性表达率为12．5％（1／8），RT-PCR阳性率为25．0％（2／8）。另外，Survivin及VEGF在不良组织型肾母细胞瘤组织及瘤旁组织中表达均明显增高。Survivin及VEGF阳性者2年存活率均低于其表达阴性者。结论Survivin，VEGF表达与肾母细胞瘤的临床病理学特点有关，其与肿瘤血管的生成，血管内皮细胞的凋亡，稳定性密切相关，进而影响肿瘤的侵袭力。其检测有利于肾母细胞瘤的诊断及其预后情况的评价。     

Allergic factors associated with the development of asthma and the influence of cetirizine in a double-blind, randomised, placebo-controlled trial: First results of ETA®

There is a common progression known as the allergic march from atopic dermatitis to allergic asthma. Cetirizine has several antiallergic properties that suggest a potential effect on the development of airway inflammation and asthma in infants with atopic dermatitis. Methods. Over a two year period, 817 infants aged one to two years who suffered from atopic dermatitis and with a history of atopic disease in a parent or sibling were included in the ETAC^® (Early Treatment of the Atopic Child) trial, a multi-country, double-blind, randomised, placebo-controlled trial. The infants were treated for 18 months with either cetirizine (0.25mg/ kg b.i.d.) or placebo. The number of infants who developed asthma was compared between the two groups. Clinical and biological assessments including analysis of total and specific IgE antibodies were performed. Results. In the placebo group, the relative risk (RR) for developing asthma was elevated in patients with a raised level of total IgE (≥ 30 kU/I) or specific IgE (≥ 0.35 kUA/I) for grass pollen, house dust mite or cat dander (RR between 1.4 and 1.7). Compared to placebo, cetirizine significantly reduced the incidence of asthma for patients sensitised to grass pollen (RR = 0.5) or to house dust mite (RR = 0.6). However, in the population that included all infants with normal and elevated total or specific IgE (intention-to-treat - ITT), there was no difference between the numbers of infants developing asthma while receiving cetirizine or placebo. The adverse events profile was similar in the two treatment groups. Discussion. Raised total IgE level and raised specific IgE levels to grass pollen, house dust mite or cat dander were predictive of subsequent asthma. Cetirizine halved the number of patients developing asthma in the subgroups sensitised to grass pollen or house dust mite (i.e. 20% of the study population). In view of the proven safety of the drug, we propose this treatment as a primary pharmacological intervention strategy to prevent the development of asthma in specifically sensitised infants with atopic dermatitis.     

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孤独症谱系障碍(autistic-spectrum disorders,ASDs)近年来患病率逐年攀升至1%左右,其症状往往伴随终生,成为严重威胁儿童健康和发展的神经发育性疾患;注意缺陷多动障碍(attention deficit hyperactivity disorder,ADHD)是儿童期最常见的精神障碍,国内报道患病率为4.13%～5.83%,其症状可延续至青少年期,甚至到成年期[1]。这两类精神障碍在成年期的临床表现、共患病、治疗策略和预后与儿童期有哪些不同呢?本文通过回顾相     

Infiltrations of Epstein-Barr virus-carrying cells in granular lymphocyte-proliferative disorders corresponding to chronic active Epstein-Barr virus infection

A 21-year-old man with granular lymphocyte-proliferative disorders (GLPD) associated with chronic active Epstein-Barr virus (EBV) infection is described. Chromosomal analyses revealed several clonal abnormalities and two of them were mainly repetitious. High copy numbers of monoclonal EBV genome were also detected in the proliferative large granular lymphocytes (LGLs), indicating the monoclonal expansion of EBV-infected LGLs. The patient had an indolent course for several years, and there was no evidence of infiltrations of his bone marrow until the end stage. At autopsy, microscopic studies revealed marked infiltrations of LGL in the liver and spleen, and the infiltrating cells were NK-cell immunophenotype. The infiltrated LGLs showed latency I.     

LUTEINIZING HORMONE RECEPTOR MUTATIONS IN DISORDERS OF SEXUAL DEVELOPMENT AND CANCER

Human male sexual development is regulated by chorionic gonadotropin (CG) and luteinizing hormone (LH). Aberrant sexual development caused by both activating and inactivating mutations of the human luteinizing hormone receptor (LHR) have been described. All known activating mutations of the LHR are missense mutations caused by single base substitution. The most common activating mutation is the replacement of Asp-578 by Gly due to the substitution of A by G at nucleotide position 1733. All activating mutations are present in exon 11 which encodes the transmembrane domain of the receptor. Constitutive activity of the LHR causes LH releasing hormone-independent precocious puberty in boys and the autosomal dominant disorder familial male-limited precocious puberty (FMPP). Both germline and somatic activating mutations of the LHR have been found in patients with testicular tumors. Activating mutations have no effect on females. The molecular genetics of the inactivating mutations of the LHR are more variable and include single base substitution, partial gene deletion, and insertion. These mutations are not localized and are present in both the extracellular and transmembrane domain of the receptor. Inactivation of the LHR gives rise to the autosomal recessive disorder Leydig cell hypoplasia (LCH) and male hypogonadism or male pseudohermaphroditism. Severity of the clinical phenotype in LCH patients correlates with the amount of residual activity of the mutated receptor. Females are less affected by inactivating mutation of the LHR. Symptoms caused by homozygous inactivating mutation of the LHR include polycystic ovaries and primary amenorrhea.     

EXCITING NEW TREATMENT APPROACHES FOR PATHYPHYSIOLOGIC MECHANISMS OF SICKLE CELL DISEASE

During the past several decades, our understanding of the complex pathophysiology of vasoocclusion associated with sickle cell disease has improved greatly. Interaction of genes, hemoglobin molecules, red cell membrane and metabolic changes, cell-cell interactions and cell-plasma interactions, red cell adhesion to vascular endothelium, activation of coagulation, and vascular reactivity play a role in vaso occlusion. Penicillin prophylaxis of pneumococcal infections and appropriate use of blood transfusions and other supportive measures improved survival of sickle cell patients. Hydroxyurea made a major impact on sickle cell therapy when it was shown to decrease acute painful episodes, acute chest syndrome, and the need for blood transfusion in adults. Significant experience in the use of hydroxyurea has been accumulated in older children. The benefits and risks of hydroxyurea for younger children and long-term risks in all patients will be evaluated in future investigations. Other promising therapies include butyrate compounds, clotrimazole, magnesium supplementation, poloxamer 188, antiadhesion agents, anticoagulant approaches, and nitric oxide. Hemopoietic transplantation remains the only curative therapy. However, several transgenic mouse models are available for studies of gene therapy or other treatment approaches on biochemical, cellular, and pathologic effects of mutant genes.     

Bibliometric data: a disaster for many non-American biomedical journals

Bibliometric data published by the Institute of Scientific Information in Philadelphia (ISI), and which was previously discussed in Acta Paediatrica , has increasingly been used despite all the relevant and severe criticism that has been raised against this method of evaluating individual research results and grading scientific journals. It is obvious that the present trend regarding the use of bibliometric data as a basis for priorities and funding of research and for the promotion of individual scientists favours American-oriented research projects at the expense of those that are based on concepts of predominantly European relevance.

Conclusion: For the future of non-American research, it is important that no single super-power, i.e. the USA, should dominate scientific priorities. The condition for efficient European competition is that European Centres with high levels of competence for creative research and training of scientists from all over the world are established. In addition, it is important that the results of European research are published in prestigious European journals, as was the situation before World War II.     

Personality profiles and heart rate variability (vagal tone) in children with recurrent abdominal pain

The aim of the study was to explore psychological factors and autonomic activity in children with recurrent abdominal pain and to compare them with those in a control group of healthy children. The Personality Inventory for Children was used for assessment of developmental, emotional and psychosocial factors in 25 children with recurrent abdominal pain (age, 7-15 y). Parasympathetic and sympathetic functions in these children and in 23 healthy control subjects (age, 7-13 y) were also investigated, non-invasively using a computerized polygraph. Vagal tone (parasympathetic function) was indexed by calculation of respiratory sinus arrhythmia in beats/min. Skin conductance (sympathetic function) was recorded by the constant current method. On the Personality Inventory for Children, 16 patients had high scores on somatic concern. Several patients had scores in the clinical range for depression, withdrawal and anxiety, but the mean scores for these personality profile scales were well within the normal range of healthy children. Interestingly, there was a spike on the L (Lie)-scale for most of the patients and 15 patients had scores above or close to the clinical cut-off value. As compared with the scores in healthy children, vagal tone and sympathetic tone were normal. Conclusion: Many children with recurrent abdominal pain have scores in the clinical range for depression, withdrawal, anxiety and L-scale indicating coping problems, denial and a trend towards somatic concern that may contribute to the evolution of abdominal pain. Autonomic nerve activity was not disturbed in these children.     

Understanding infant formula

The World Health organisation recommends breast feeding infants for the first six months of life. When this breast feeding does not occur either through parental choice or medical need, infant formulas will be required. There is a bewildering array of formulas on the UK market for many different requirements. When faced with an unsettled infant many parents (and healthcare professionals) will experiment with the infant formula available and then attend the paediatric clinic looking for help and advice. It is therefore essential that paediatricians understand what milks are available and what the key differences between different products are. This review attempts to provide a simple guide through many of the formulations currently available in the UK; and offers advice for the dietary management of the child with extra calorie requirements, infants with cow's milk protein allergy, gastro oesophageal reflux disease, apparent unresolved hunger and infantile colic. Whatever the underlying condition, there is likely to be an infant formula that is suitable in this generation of ever expanding formulations.     

Overcoming surfactant inhibition with polymers

Inhibition of the function of pulmonary surfactant in the alveolar space is an important element of the pathophysiology of many lung diseases, including meconium aspiration syndrome, pneumonia and acute respiratory distress syndrome. The known mechanisms by which surfactant dysfunction occurs are (a) competitive inhibition of phospholipid entry into the surface monolayer (e.g. by plasma proteins), and (b) infiltration and destabilization of the surface film by extraneous lipids (e.g. meconium-derived free fatty acids). Recent data suggest that addition of non-ionic polymers such as dextran and polyethylene glycol to surfactant mixtures may significantly improve resistance to inhibition. Polymers have been found to neutralize the effects of several different inhibitors, and can produce near-complete restoration of surfactant function. The anti-inhibitory properties of polymers, and their possible role as an adjunct to surfactant therapy, deserve further exploration.