MYCN基因监测在儿童恶性实体瘤中的临床研究进展

恶性实体瘤在儿童中发病率不高,却是儿童主要死亡原因.MYCN基因扩增是神经母细胞瘤的独立的不良预后因素,参与神经母细胞瘤肿瘤形成,是分层治疗的重要依据.同时MYCN基因扩增在其他儿童恶性实体瘤中亦可检测到,并与髓母细胞瘤的不良病理表现、肾母细胞瘤和肺泡型横纹肌肉瘤的不良预后有关.该文比较MYCN基因的各种检测方法,总结其与儿童实体瘤的临床关系,探讨分子特异性治疗提高儿科实体瘤治愈率的可能.     

CIE及IEV方案在恶性实体瘤儿童自体外周血干细胞动员和采集中的应用

目的：自体外周血干细胞移植（APBSCT）是治疗儿童恶性实体瘤的重要方法之一,干细胞动员与采集是决定造血重建的重要因素。本研究采用CIE及IEV动员方案对儿童神经母细胞瘤（NB）及横纹肌肉瘤进行干细胞动员和采集,并对临床效果进行评价。方法：8例IV期NB 患儿采用CIE化疗方案动员,3例III期横纹肌肉瘤患儿采用IEV方案。观察采集干细胞的效果。结果：11例患儿平均采集单个核细胞数（MNC）为(5.55 ±1.43)×108/kg,CD34+ 细胞数为（4.88±2.48）×106/kg,动员并发症少,患儿均能耐受。10例行APBSCT后均获快速造血功能重建,其中1例NB于APBSCT后32 d因合并左心衰竭死亡,余9例患儿APBSCT后60 d复查外周血象稳定。结论：CIE及IEV方案可以安全有效地完成NB及横纹肌肉瘤自体外周血干细胞动员和采集并达到移植要求。     

儿童噬血细胞综合征32例临床分析

目的 探讨儿童噬血细胞综合征(hemophagocytic syndrome,HPS)的临床表现、病因、诊断及治疗.方法 回顾性分析我科收治的32例HPS患儿的病因、临床症状、体征、实验室检查结果及治疗转归情况.结果 (1)病因:全部病例中病毒感染相关性HPS占75.0％ (24/32),其中以EB病毒感染最为常见,占68.8％ (22/32).(2)临床表现:主要为持续发热,肝、脾及淋巴结肿大.(3)实验室检查特点:外周血常规表现为三系或二系减少,血清铁蛋白明显增高,肝功能受损,三酰甘油升高和(或)纤维蛋白原降低,有凝血障碍.骨髓涂片可找到噬血细胞,可溶性白细胞介素-2受体增高,自然杀伤细胞活性降低.(4)治疗及转归:32例HPS患儿中,24例病毒感染相关性HPS患儿均予抗病毒药物、大剂量人血丙种球蛋白治疗.22例予糖皮质激素、环磷酰胺等治疗,好转11例,完全缓解5例,死亡4例,疗效不佳自动出院2例;另外10例采用HLH-2004方案治疗,好转4例,完全缓解6例.结论 HPS多由感染尤其是EB病毒感染所诱发,血清铁蛋白、可溶性白细胞介素-2受体及自然杀伤细胞活性可早期反映疾病转归,化疗过程中应定期监测.HLH -2004方案是HPS有效的治疗方案.     

Phase I trial of subcutaneous interleukin-1α in children with malignant solid tumors

Interleukin-1α (IL-1α) is myeloprotective in a variety of animal models of cancer chemotherapy and is similarly beneficial in adults treated with carboplatin, 5-fluorouracil, and after autologous bone marrow transplantation. There are no trials of this agent in children. Our purpose was to determine the toxicity and maximum tolerated dose (MTD) of recombinant human interleukin-1α (rhuIL-1α) in children with solid tumors receiving intensive cancer chemotherapy and to evaluate its myelo-protective effects. Cohorts of patients received rhuIL-1α in doses of 0.1–10 μg/m² for 4 days by subcutaneous injection prior to ICE chemotherapy (ifosfamide, 2 g/m²/day × 3, carboplatin targeted to an area under the curve of 8 mg/ml × min on day 1, and etoposide, 100 mg/m² daily for 3 days). Patients were randomized to receive rhuIL-1α before either the first or second course of therapy. After the MTD of rhuIL-1α was determined, an additional group of patients received rhuIL-1α at that dose immediately following ICE chemotherapy. The dose-limiting toxicities of rhuIL-1agr; in the 27 children tested comprised systemic symptoms of fever, chills, headache, and hypotension. The MTD was 3 μg/m²/day. There were no differences in chemotherapy-induced hematologic toxicity with increasing doses of rhuIL-1α or in comparisons before or after ICE chemotherapy. Although rhuIL-1α can be given safely to children receiving myelosuppressive chemotherapy, clinical usefulness would mandate a significant hematopoietic benefit in view of the troublesome side effects identified. We saw no evidence of a hematoprotective effect. Med. Pediatr. Oncol. 28:444–450, 1997. © 1997 Wiley-Liss, Inc.     

The role of neoadjuvant chemotherapy in children with malignant solid tumors

Pediatric surgeons play a critical role in diagnosing, staging, and treating malignant solid tumors in children. Over the years, the surgical management of the primary tumor site has evolved from an aggressive en-bloc resection at diagnosis to a more tailored surgical approach, often affecting definitive local control after the delivery of neoadjuvant therapy, as currently directed by many solid tumor protocols. In fact, inappropriate upfront resection can lead to unnecessary short- and long-term morbidity, an incomplete resection, and may be associated with a delay in the initiation of the systemic chemotherapy that is critical to the treatment of gross or occult metastatic disease. Therefore, it is important for the pediatric surgeon, as a member of the multidisciplinary team involved in the care of these children, to understand the indications for and implications of neoadjuvant therapy in the treatment of pediatric solid tumors. Here we review the current management of childhood solid tumors focusing on the role of neoadjuvant therapy.     

Defective natural killer cell activity and deficient production of interferon-γ in children with acute lymphoblastic leukemia

Natural killer (NK) cell activity, OK-432-augmented-NK cell activity, concentrations of interferon-γ (IFN-γ) in the culture supernatants of lymphocytes stimulated with OK-432, and subsets of NK cells and memory T cells were analyzed in 42 children with acute lymphoblastic leukemia (ALL) receiving maintenance chemotherapy. Natural killer and augmented-NK cell activities, and concentrations of IFN-γ in the supernatants of cultured lymphocytes, were significantly lower in the patients with ALL than in age-matched control children. Among the NK cell subsets, proportions of CD57⁺ cells in the patients with ALL were significantly higher than in the controls, and proportions of a memory T cell subset (CD4⁺ CD29⁺ T cells) in the patients were also significantly higher than in the controls. These results suggest that the function of NK cells and memory T cells that are considered as IFN-γ producing cells, may be defective in ALL, and that CD57⁺ cells and CD4⁺ CD29⁺ cells may be resistant to or recover rapidly from suppression by cytotoxic chemotherapy.     

自然杀伤细胞与丙型肝炎病毒感染的Huh7.5细胞体外共培养后的功能变化

目的探讨体外丙型肝炎病毒(HCV)感染对NK细胞功能产生的影响及其可能的机制。方法利用质粒JC1-Flag2体外转录得到的HCV感染性颗粒(HCVcc)以MOI4.8感染Huh7.5细胞(Huh7.5-HCVcc),与健康人外周血分离得到的NK细胞进行共培养。与Huh7.5-HCVcc细胞共培养前后,采用ELISA法和MTT比色法分别检测NK分泌细胞因子IFN-γ、TNF-α和IL-10的水平和细胞杀伤活性,以评估HCV感染对NK细胞功能的影响。结果与MOI4.8的Huh7.5-HCVcc细胞共培养,NK细胞分泌IFN-γ、TNF-α和IL-10水平受到抑制,其中IFN-γ在共培养6 h(P0.001)、9 h(P0.001)、12 h(P0.001)受到显著抑制,TNF-α在共培养6 h(P0.001)、9 h(P0.001)、12 h(P=0.001)受到显著抑制,IL-10在共培养6 h(P0.001)、9 h(P=0.006)受到显著抑制;且NK细胞分泌细胞因子IFN-γ、TNF-α和IL-10的功能均在共培养6 h受到的抑制效应最大,平均抑制率分别为24.1%、20.7%和24.3%。NK细胞杀伤活性在共培养6 h(P=0.023)受到抑制,平均抑制率为16.6%。结论在体外与MOI4.8的Huh7.5-HCVcc细胞共培养,NK细胞分泌细胞因子(IFN-γ、TNF-α和IL-10)和细胞杀伤功能均受到抑制,且在不同时间点受到的抑制程度不同,提示NK细胞在HCV感染的不同阶段可能起到不同的作用。     

自体外周血造血干细胞移植在小儿恶性晚期实体肿瘤的临床应用(英文)

目的　小儿晚期实体肿瘤对常规化疗效果欠佳 ,该文探讨大剂量化疗并自体外周血干细胞移植(APBSCT)治疗小儿高危晚期实体瘤的可行性及疗效。方法　 1 3例恶性实体肿瘤患儿 (恶性淋巴瘤 7例、神经母细胞瘤 6例 ) ,在其完全缓解 (1 2例 ) ,部分缓解 (1例 )后进行了APBSCT治疗。移植时病程中位时间 1 0月。 1 1例用化疗加重组人粒 单细胞集落刺激因子 (rhGM CSF)或重组人粒细胞集落刺激因子 (rhG CSF)动员 ,2例采用常规化疗方案作为动员剂。所采集单个核细胞 (MNC)为 (6 .85± 2 .6 5 )× 1 0 8/kg。CD34+ 细胞为 (1 5 .82± 1 2 .93)×1 0 6/kg。CFU GM集落为 (1 7.87± 1 7.94 )个 / 1 0 4细胞。预处理方案中 6例基本方案为全身放疗加环磷酰胺。 7例未用TBI ,仅以马法兰为主做为预处理方案 (马法兰 +卡铂 +足叶乙甙 5例 ,白消胺 +马法兰 2例 )。结果　移植后白细胞 >0 .5× 1 0 9/L、>1 .0× 1 0 9/L、血小板 >2 0× 1 0 9/L的中位时间分别为 1 2天、1 5天、1 9天。中位随访时间4 8月 (1月～ 1 4 4月 )。至今总生存率 77% (1 0 / 1 3) ,死亡率 2 3% (3/ 1 3) ,无移植相关死亡。结论　APBSCT是治疗小儿晚期实体肿瘤 ,明显改善其预后的重要治疗方法。     

神经母细胞瘤并骨转移患儿临床特征及预后相关因素的单中心分析CSCD

目的分析伴骨转移神经母细胞瘤(neuroblastoma,NB)患儿的临床特征及预后相关因素,总结临床诊疗经验,以提高伴骨转移NB患儿的生存率。方法以2013年12月至2020年12月重庆医科大学附属儿童医院肿瘤外科收治的伴骨转移NB患儿为研究对象,收集并总结患儿临床资料及预后情况。随访时间截至2021年3月31日。结果共收集97例NB患儿,男68例,女29例,男女比例为2.4∶1;中位年龄为49.4个月。首发症状:发热47例(48.5%),骨痛38例(39.2%),腹痛或腹胀29例(29.9%),咳嗽15例(15.5%);伴骨转移的NB患儿存在多类骨转移(62.9%),且合并骨髓转移(73.2%),整体预后差,1年生存率为93.6%,5年生存率仅20.2%。经生存分析发现,女性、伴多类骨转移、肿瘤位于腹部、首诊时LDH测定值大于660 U/L以及术中肿瘤残留是预后不佳的影响因素,其中手术切除范围是独立预后影响因素。结论伴骨转移的NB患儿临床表现多样,其预后受诸多因素影响,肿瘤复发或进展是主要的致死原因。对于此类患儿,建议术中尽可能完全切除肿瘤,以改善预后。     

神经母细胞瘤并骨转移患儿临床特征及预后相关因素的单中心分析

目的 分析伴骨转移神经母细胞瘤(neuroblastoma,NB)患儿的临床特征及预后相关因素,总结临床诊疗经验,以提高伴骨转移NB患儿的生存率.方法 以2013年12月至2020年12月重庆医科大学附属儿童医院肿瘤外科收治的伴骨转移NB患儿为研究对象,收集并总结患儿临床资料及预后情况.随访时间截至2021年3月31日...     

B19 parvovirus infection in children with malignant solid tumors receiving chemotherapy

Two children with rhabdomyosarcoma developed severe anemia following chemotherapy; anemia was more severe compared to that observed following earlier chemotherapy cycles. While one patient had a brisk reticulocytosis, the other had no demonstrable reticulocytes. Both patients had evidence of acute B19 parvovirus infection and subsequently developed appropriate antibody response. A diagnosis of B19 parvovirus infection should be considered in any patient who develops persistent or severe anemia while on chemotherapy. © 1994 Wiley-Liss, Inc.     

IL-12对支气管哮喘小鼠Th2免疫应答的影响

目的　哮喘是Th2细胞介导的气道慢性非特异性炎症。IL 12在抗原致敏阶段可有效抑制Th2免 疫应答形成,而对已产生的Th2免疫应答是否同样具有抑制作用尚有争议。本研究探讨IL 12对哮喘时已产生的 Th2免疫应答的影响,为临床应用提供可靠的理论依据。方法　实验一:取25只6～8周雌性BALB/c小鼠,腹腔 注射鸡卵清蛋白(OVA)和氢氧化铝致敏。于致敏前、致敏后第7,14,21,28天,各取处死5只检测脾脏单个核细胞 培养上清液中的Th2类细胞因子IL 4和IL 5水平,观察Th2免疫反应形成情况。实验二:另取40只相同小鼠随机 分为哮喘组(n=20)和IL 12治疗组(n=20),均腹腔注射鸡卵清蛋白(OVA)和氢氧化铝致敏后雾化吸入OVA诱 发哮喘。IL 12治疗组在致敏后25d开始连续5d腹腔内注射0.5μg(1mL)重组IL 12,哮喘组仅注射PBS。两组 均在最后1次诱发后(致敏后30d)处死,取支气管肺泡灌洗液(BALF)和脾脏单个核细胞培养上清液检测Th2类 细胞因子IL 4、IL 5和Th1类细胞因子IFN γ水平(ELISA法)。结果　实验一:小鼠在致敏后14d脾脏单个核细 胞分泌IL 4和IL 5明显增高,说明小鼠致敏后第14天已形成Th2免疫应答。实验二:与哮喘组比较,IL 12治疗组 BALF中IL 4(192±19pg/mLvs19±5pg/mL)和IL 5浓度(328±71pg/mLvs141±15pg/mL     

Analysis of Killer Cell Activities in Epstein-Barr Virus Infections

Using spontaneously established autologous lymphoblastoid B cell lines (LCL), killer cell activities were studied in children with severe infectious mononucleosis (IM), chronic IM, and acute IM, and compared with those in EBV-seropositive normal controls. Natural killer (NK) cell activity of fresh peripheral blood mononuclear cells (PBMC) was normal in acute IM patients, but it was low in four of six patients with severe or chronic IM. Recombinant inter-leukin 2 (rIL-2)-activated PBMC from normal controls showed lymphokine-activated killer (LAK) cell activity against the respective autologous LCL. The levels of LCL lysis by LAK cells were significantly higher in acute IM patients, lower in chronic IM patients, and much lower in severe IM patients. In contrast to the fact that PBMC stimulated in vitro with autologous LCL (IVS cells) from normal controls and acute IM patients showed potent killing of autologous LCL, IVS cells from severe or chronic IM patients showed lower levels of LCL lysis, which were markedly augmented in three patients by rIL-2 addition to the cultures. These killer cell dysfunctions appear to be responsible for the severe or chronic course of EBV infection.     

自身造血干细胞移植治疗儿童晚期恶性实体肿瘤

目的　评价自身干细胞移植治疗儿童晚期恶性实体肿瘤的安全性及疗效。方法13例次患儿直接采集骨髓 ,另 15例患儿经粒细胞集落刺激因子 (G CSF)动员后从外周血获取单个核细胞 ,对 1例疑有肿瘤细胞浸润的采集物经CliniMACS进行了CD 3 4 细胞分选的净化处理。除 2例霍奇金淋巴瘤患儿经CBV方案 (CTX BCNU VP16 ) (环磷酰氨、卡氮芥及依托泊甙 )治疗外 ,其余患儿均采用VP16 卡铂 马法兰的预处理方案。结果　采集骨髓及外周血得到的单个核细胞分别为 ( 5 4± 2 1)× 10 8/kg和 ( 4 1± 1 9)× 10 8/kg。所有患儿移植后均获得造血重建 ,中性粒细胞恢复至 0 5×10 9/L的时间为 ( 11 8± 5 7)d ,血小板大于 2 0× 10 9/L的时间为 ( 2 1 0± 9 3)d。移植过程中 3例患儿分别合并表皮葡萄球菌、腐生葡萄球菌和枯草杆菌败血症 ,无一例因移植相关并发症而死亡。但 1例出现急性肾功能不全、肺水肿、心包积液并发展为呼吸窘迫综合征 ,经机械通气、应用肺表面活性物质等积极治疗后康复 ;另 1例出现BCNU相关的肺损伤 ,导致严重的肺动脉高压、嗜酸性细胞增多 ,经皮质激素等治疗后逐渐好转。本组平均随访 13个月 ,2 7例患儿中 5例移植后 5个月内因疾病复发而死亡 ;1例非霍奇金淋巴瘤患儿移植后 3个月中枢神经系统复     

Killing of human Herpes virus 6-infected cells by lymphocytes cultured with interleukin-2 or -12

BACKGROUND: Human Herpes virus 6 (HHV-6) is a causative agent of exanthema subitum and replicates mainly in lymphocytes. The aim of present study was to investigate cytotoxicity against HHV-6-infected cells by cord blood mononuclear cells (CBMC) and adult peripheral blood mononuclear cells (PBMC). METHODS: Human herpes virus 6-infected and -uninfected lymphocytes were used as target cells. Killing of target cells by CBMC and PBMC was investigated by the chromium release cytotoxicity assay. RESULTS: Freshly isolated CBMC and PBMC did not lyse HHV-6-infected and -uninfected cells. When CBMC and PBMC were cultured with interleukin (IL)-2, HHV-6-infected cells were significantly lysed compared with uninfected cells. Deletion of CD16+ cells by treatment of effector cells with anti-Leu-11b (CD16) antibody with complement reduced cytotoxicity against HHV-6-infected cells and T lymphocyte-rich cells did not lyse HHV-6-infected cells. Treatment of effector cells with anti-Fas ligand antibody and treatment of HHV-6-infected cells with anti-Fas antibody reduced cytotoxicity against HHV-6-infected cells. DNA fragmentation was detected in the supernatant from HHV-6-infected cells cultured with IL-2-activated lymphocytes. Culture of CBMC and PBMC with IL-12 also enhanced cytotoxicity against HHV-6-infected cells. CONCLUSIONS: These data suggest that lymphocytes cultured with IL-2 or IL-12 mediate killing against HHV-6-infected cells and killing of HHV-6-infected cells was through apoptosis. Fas-Fas ligand interaction is one pathway by which HHV-6-infected cells are killed. Killing of HHV-6-infected cells by NK cells activated by cytokines may play a role in the recovery from HHV-6 infection in vitro.     

儿童恶性实体瘤远期生存质量评估研究进展

近年来,我国部分地区报道儿童恶性肿瘤的发病率逐渐增高~([1-2]),其中很大部分是儿童恶性实体瘤患者.儿童常见的恶性实体肿瘤包括中枢神经系统肿瘤、神经母细胞瘤、肾母细胞瘤、横纹肌肉瘤、尤文肉瘤、生殖细胞瘤、骨肉瘤、视网膜母细胞瘤、肝母细胞瘤等.