Cell proliferation in explant cultures of malignant gliomas

Summary Primary explant cultures were grown from human malignant gliomas. Ten-day cultures contained pleomorphic tumour cells with overlapping processes and contaminating elements, consisting chiefly of fibroblasts which grew in sheets of contact-inhibited cells. Cultures pulse-labelled with thymidine-H³ for one hour showed labelling in many fibroblast nuclei. Virtually no labelling was seen in the cells of apparent tumour origin. Most labelled cells were concentrated near the explant. Results are discussed in relation to the mode of outgrowth of the different cell types.     

E1B缺陷性腺病毒对人恶性脑胶质瘤的溶瘤作用

目的 研究E1B缺陷性腺病毒d11520对人恶性脑胶质瘤细胞株U251的体外溶瘤作用. 方法 按感染复数(MOI)为50、5、0.5、0.05、0.005、0 pfu将E1B缺陷性腺病毒dl1520感染常规培养的U251、Hep3B、T24细胞(Hep3B为阳性对照,T24为阴性对照),结晶紫染色观察细胞病变(CPE)出现的时间:MOI为5pfu时病毒空斑试验检测dl1520在3种细胞中的复制.将携带报告基因βgal的腺病毒载体Ad-βgal感染细胞,计算dl1520对3种细胞的感染率. 结果 结晶紫染色结果显示Hep3B细胞对dl1520最敏感、发生CPE效应最快,U251次之,而T24则不产生CPE效应；dl1520在U251细胞中复制数和感染率均低于Hep3B细胞,但明显高于T24细胞,差异均有统计学意义(P＜0.05). 结论 E1B缺陷性腺病毒dl1520对人恶性脑胶质瘤细胞具有显著的溶瘤作用.     

Specific chromosomal abnormalities characterize four established cell lines derived from malignant human gliomas

Summary The four permanent human glioma-derived cell lines reported here are the first such lines for which the karyotypes have been followed from the original biopsies through the establishment of the lines in culture. Although ploidy changes were seen, each line retained either distinctive marker chromosomes or the overall original chromosomal distribution allowing the origin of each line to be established with certainty. D-263 MG expresses glial fibrillary acidic protein, all lines except D-245 MG are tumorigenic in athymic mice, and each line displays a unique pattern with respect to in vitro growth parameters and expression of biochemically defined markers, oncofetal antigens and lymphoid-associated markers. D-245 MG and D-259 MG are able to grow in the absence of supplemental glutamine; glutamine synthetase was detected in these cell lines both by immunocytochemistry and by direct assay. Thus, the four permanent human glioma-derived cell lines described here are representative of glioma lines in their general characteristics. D-259 MG retains numerous double minute chromosomes (DMs), D-263 MG contains two marker chromosomes with breaks in 9p, and D-247 MG and D-245 MG with stemlines containing 96 and 89 chromosomes contain eight and six normal copies (respectively) of chromosome No. 7. The retention in these four cell lines of the most common chromosomal abnormalities seen in biopsies of malignant human gliomas provides the opportunity to investigate the meaning of these specific chromosomal changes.Dedicated to Prof. F. Seitelberger on the occasion of his seventieth birthdaySupported on part by P01 NS0023 from the National Institute of Neurological and Communicative Disease and Stroke, R01 CA11898 from the National Cancer Institute and The Swedish Cancer Society     

伽玛刀治疗胶质瘤的近期效果分析

目的 探讨不同级别的脑胶质瘤伽玛刀治疗后的近期效果。方法自2002年6月至2004年8月对病理诊断明确的97例脑胶质瘤患者进行了伽玛刀治疗,随访66例,其中位于颞叶10例,额叶16例,顶叶11例,枕叶6例,丘脑9例,小脑6例,脑干5例,视交叉3例,体积在3～32cm^3之间,平均10.38cm^3。29例病人伽玛刀治疗前经开颅手术,大多数肿瘤的边缘剂量为12～16Gy(6～25Gy),相应的中心剂量为25～30Gy(14～55Gy)。结果平均随访时间为8．1个月(3～24个月),经MRI或CT复查．治疗后3～6个月,28．8％肿瘤体积减小,42.4％肿瘤停止生长,21．2％肿瘤继续增大,死亡5例,占7．7％。25.8％患者肿瘤周围出现不同程度的水肿,19．7％的患者原来的瘤周水肿减轻或消失。结论伽玛刀是治疗脑胶质瘤的一种有效的方法。     

Disialoganglioside GD2 in human neuroectodermal tumor cell lines and gliomas

Summary Monoclonal antibodies (mAbs) recognizing the disialoganglioside II³(NeuAc)₂GgOse₃Cer (GD2) were produced by immunizing mice with the GD2-expressing neuroblastoma cell line LAN-1 and a prefusion boost with purified GD2 coupled to Salmonella minnesota. Two IgM mAbs were isolated which demonstrated high levels of reactivity (binding ratios in excess of 100) with GD2 by solid-phase radioimmunoassay and positivity in high-performance thin-layer chromatography (HPTLC) immunostain; only one (DMAb-20) was subsequently shown by analysis with a panel of defined ganglioside species to be specific for the minimum epitope of GD2, GalNAc1-4(NeuAc2-8NeuAc2-3)Gal-. DMAb-20 was used to evaluate the expression of GD2 by malignant glioma and medulloblastoma cell lines using cell surface radioimmunoassay, indirect membrane immunofluorescence, HPTLC immunostain, and densitometric analysis of extracted gangliosides from selected cell lines. Sixteen of 20 (80%) malignant glioma and 5 of 5 medulloblastoma cell lines reacted with DMAb-20; in agreement with previous studies, 5 of 5 neuroblastoma and 2 of 3 melanoma cell lines also reacted with DMAb-20. GD2 was proportionally increased in the glioma and medulloblastoma cell lines relative to levels in normal brain, as determined by densitometric analysis. In a phenotypic survey of malignant glioma biopsies, tumor cells in 24 of 30 (80%) cases stained positively with DMAb-20. Reactive astrocytes, both within and adjacent to tumors, were frequently intensely stained. Among the morphological variants of glioblastoma examined, the most intense staining with DMAb-20 was observed in neoplastic gemistocytes, with the weakest or absent staining in small cell glioblastomas. As GD2 is a commonly expressed surface antigen of gliomas and medulloblastomas, expression of which is retained in tissue culture, DMAb-20 will be useful in determining the functional role of GD2 in cell-cell interaction, adhesion, and invasion, and in defining altered growth control mechanisms of central nervous system neoplasms in in vitro models.Abbreviations xxGangliosides have been designated according to CBN recommendations [22] and to the coding system of Svennerholm [35] GD2 II³(NeuAc)₂GgOse₃Cer - GM3 II³NeuAc-LacCer - GD3 II³(NeuAc)₂-LacCer - GM2 II³NeuAcGgOse₃Cer - GM1 II³NeuAcGgOse₄Cer - GD1a II³NeuAcIV³NeuAcGgOse₄Cer - GD1b II³(NeuAc)₂GgOse₄Cer - GT1b IV³NeuAcII³(NeuAc)₂GgOse₄Cer - GQ1b IV³(NeuAc)₂II³(NeuAc)₂GgOse₄Cer - 3,8-LD1 IV³(NeuAc)₂nLeOse₄Cer Supported in part by NIH Grants R37 CA 11898, NS 20023, CA 32672, and T32-NS 07304, and by a grant from the Swedish Medical Research Council (Project 03X-627). Dr. Longee is an Association of Medical School Pediatric Department Chairmen, Inc., Pediatric Scientist Training Program Fellow supported by St. Jude Children's Research Hospital     

The chromosomal aberraation of double-minutes in three gliomas

Summary Thkek chromosomes of about 40 human malignant gliomas in adults have been studied. Three tumours had double-minutes (dms) in all or most of their cells whereas 5 additional gliomas showed the same aberration in sporadic cells. The humber of dms varied between 1 and more than a hundred. In most case the majority of the double-minutes were extremely smasll and they were morphologically similar to those seen in some murine sarcomas. Only a few of the dms seemed to be centric. The occurrence of dms could not be related to any particular deviations as regards ordinary chromosomes. Some observations suggested that the dms could be derived drom one or a few pulverized chromosomes. The period the dms could remain in the tumour cell population is probably limited. This problem is discussed together with a possible mechanism for their initial increase in number in the tumour cells.The present work was supported by grants form the Sweedish Cancer Socitety, the University of Lund, and John and Augusta Persson's foundation for medical reseaches     

应用神经导航系统对成人幕上胶质瘤手术切除的临床研究

目的 应用神经导航系统对成人幕上胶质瘤实施手术，研究肿瘤全切除与神经功能保护。方法 85例病人，病变位于大脑半球各部位，为不同病理类型的胶质瘤。肿瘤直径从1．8cm到7．0cm，平均5．5cm。手术全过程在导航系统引导与监视下进行。结果 肿瘤全切76例，近全切5例，大部切除4例。所有肿瘤全切病人术后增强颅MRI或CT检查未发现肿瘤残留病灶。手术后有49例病人出现明显的神经功能障碍，表现为不同程度的肢体瘫痪和(或)语言障碍，不全偏盲，但全部病例在2个月内恢复，无一例病人留有永久性神经功能障碍。所有8例术前偏瘫的病人在术后短期内症状得到明显改善，均下地行走，KPS从术前的60—70分增加到80—90分。结论 应用神经导航系统可对大多数大脑半球不同部位胶质瘤病人做到影像学意义肿瘤全切除，并且不引起病人术后永久性神经功能障碍。     

MGMT表达指导恶性胶质瘤的替莫唑胺化疗(附40例报告)

目的 根据肿瘤组织O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)表达的差异选择不同的替莫唑胺(TMZ)方案对恶性胶质瘤进行化疗,评价其客观疗效、生存率和不良反应.方法 经手术后病理确诊的初治或复发恶性胶质瘤患者40例,均接受过放疗并有残留或复发可评价病灶.采用免疫组织化学方法检测肿瘤组织MGMT表达,将患者分为阳性组和阴性组.阴性组选择TMZ常规5天方案化疗,阳性组选择TMZ小剂量持续或TMZ联合顺铂方案化疗.结果 阴性组22例,阳性组18例.阳性组患者的一般状况比阴性组差,多数为复发患者且既往接受过化疗(P＜0.05).阴性组和阳性组患者的客观有效率分别为31.8%和33.3%,中位无进展生存时间分别为7个月(95%CI:5.7-8.3)和7个月(95%CI:2.3-11.7),中位生存时间分别为24个月(95%CI:12.7-35.3)和11.5个月(95%CI:9.9-11.3),差异均无统计学意义(P＞0.05).主要不良反应为Ⅰ～Ⅱ度的中性粒细胞下降(20%)、食欲下降(30%)、恶心呕吐(22%)和转氨酶升高(25%),联合顺铂方案的中性粒细胞下降发生率高于TMZ单药方案(P＜0.05).结论 TMZ小剂量持续或联合顺铂方案化疗可能改善MGMT阳性恶性胶质瘤患者的临床疗效,耐受性好.

Abstract：

Objective This study was to evaluate the efficacy and toxicity of temozolomide(TMZ)chemotherapy based on O6-methylguanine-DNA methyltransferase (MGMT) protein expression in patients with malignant gliomas.Methods A total of 40 patients with pathologically confirmed malignant gliomas were enrolled.All patients had pretreated with radiotherapy and had assessable lesions.MGMT protein expression were detected by immunohistochemical technique.Alternative schedules of TMZ were administered based on MGMT protein expression.Patients with MGMT negtive expression received 150～200 mg/m2 TMZ on days 1-5.Patients with MGMT postive expression received 75 mg/m2 TMZ on days 1-21 or 150～200 mg/m2 TMZ on days 2-6 combined with Cisplatin (DDP) 40 mg/m2 on days 1-2.Results Among 22 patients with MGMT negative expression and 18 patients with MGMT positive expression,the response rate was 31.8％ and 33.3％ (P ＞0.05 ),the median progression-free survival time was 7 months (95％ CI:5.7-8.3) and 7 months(95％ CI:2.3-11.7 ),the median survival time was 24 months(95％ CI:12.7-35.3) and 11.5 months (95％ CI:9.9-11.3 ),respectively.There were no significant differences (P>0.05 ).The most common toxicities were grade Ⅰ～Ⅱ neutropenia (20％ ),decreased appetite (30％ ),nausea and vomitting (22％ )and elevated liver enzymes (25％ ).Conclusion Alternative schedules of TMZ may improve efficacy in patients with MGMT positive expression tumors,toxicities were tolerable.     

脊髓胶质瘤治疗的进展

脊髓髓内肿瘤中脊髓胶质瘤占最重要的位置,其发病率可占脊髓髓内肿瘤的80%以上,其他类型的脊髓髓内肿瘤主要包括皮样囊肿、表皮样囊肿、肠源性囊肿、畸胎瘤、蛛网膜囊肿和血管母细胞瘤即血管网状细胞瘤等。它们的临床特点和起病形式有许多相似之处,但治疗策略和方式不尽相同,除     

儿童丘脑胶质瘤的显微外科治疗

目的 探讨儿童丘脑胶质瘤的临床特征和显微外科治疗.方法 总结49例儿童丘脑胶质瘤的临床特征,分别采取经胼胝体-穹窿间入路、胼胝体-侧脑室入路、经顶或顶枕-侧脑室(经三角区)入路、经额-侧脑室入路、经颞皮层入路、经颞下小脑幕入路进行显微外科治疗.结果 初次手术中,近全切除23例,大部切除17例,部分切除9例,围手术期死亡2例.39例获得随访,16例存活,其中15例生活可以自理.结论 手术是儿童丘脑胶质瘤的首选治疗,根据肿瘤的位置、生长方向及术者经验采取合理的手术入路,尽可能减少重要结构的损伤,减少术后并发症.     

胶质瘤中MMP-2和TIMP-2的表达及意义

目的探讨MMP-2和TIMP-2与胶质瘤侵袭性及恶性表型之间的关系及其意义。方法采用Elivision二步免疫组织化学法染色观察MMP-2和TIMP-2在46例不同恶性度胶质瘤及10例正常脑组织中的表达并用德国LeicaQ550cw图像分析系统测其灰度值作为表达强度的量化指标。结果在对照组、低度及高度恶性胶质瘤中,MMP-2的阳性表达率分别为10%、63.6%和95.8%;在对照组、低度及高度恶性胶质瘤中,TIMP-2的阳性表达率分别为10%、36.3%和37.5%。MMP-2在Ⅰ、Ⅱ级和Ⅲ、Ⅳ级胶质瘤中平均灰度值分别为173.27±13.26和98.63±18.20;TIMP-2在Ⅰ、Ⅱ级和Ⅲ、Ⅳ级胶质瘤中平均灰度值分别为210.44±12.95和205.65±9.75。结论MMP-2表达随胶质瘤恶性程度增加而增强,可作为胶质瘤恶性表型及侵袭性指标之一。TIMP-2表达在正常脑组织及不同级别胶质瘤中无明显差异。MMP-2/TIMP-2的比值与胶质瘤侵袭性密切相关。     

人脑胶质瘤的表皮生长因子基因表达

采用Ｎｏｒｔｈｅｒｎ杂交和斑点杂交的方法检测了５０例胶质瘤、４个恶性胶质瘤体外细胞系和７例正常脑组织表皮生长因子（ＥＧＦ）ｍＲＮＡ表达水平。结果显示：上述组织或细胞都可表达５．０ｋｂ的ＥＧＦｍＲＮＡ片断，高恶度胶质瘤较低恶度胶质瘤和正常脑组织ＥＧＦｍＲＮＡ表达水平增高，低恶度胶质瘤与正常脑组织的表达无显著差异。在５０例胶质瘤中２９例（５８％）ＥＧＦｍＲＮＡ过表达，其中多见于恶性胶质瘤，ＥＧＦｍＲＮＡ表达与肿瘤的分级呈正相关。提示胶质瘤可以合成ＥＧＦ，在胶质瘤的发生发展过程中，ＥＧＦ参与或促进了其恶性进展。     

血管内皮生长因子与水孔蛋白4在胶质瘤及脑转移瘤中的表达及意义

目的 探讨血管内皮生长因子(VEGF)与水孔蛋白4(AQP4)在胶质瘤及脑转移瘤中的表达,并探讨两者与胶质瘤及脑转移瘤的组织病理学关系及在瘤周水肿形成过程中的作用.方法 选择福建医科大学附属第一医院神经外科自1999年至2001年手术切除并经病理检查证实的胶质瘤石蜡组织标本73例和脑转移瘤组织标本15例,并另取正常脑组织标本8例作为对照,应用免疫组织化学方法检测组织标本中VEGF与AOP4的表达.结果 正常脑组织中未见VEGF表达:高级别胶质瘤与低级别胶质瘤之间、低级别胶质瘤与正常脑组织之间、脑转移瘤与正常脑组织及低级别胶质瘤之间VEGF阳性表达比较差异有统计学意义(P<0.05),而脑转移瘤与高级别胶质瘤之间VEGF阳性表达比较差异无统计学意义(P>0.05).AQP4在所有组织标本中均有表达,正常脑组织与高级别胶质瘤、脑转移瘤之间,低级别胶质瘤与高级别胶质瘤、脑转移瘤之间AQP4阳性表达比较差异有统计学意义(P<0.05),而正常脑组织与低级别胶质瘤之间、脑转移瘤与高级别胶质瘤之间AQP4阳性表达比较差异无统计学意义(P>0.05).Spearman相关分析显示两者在胶质瘤及脑转移瘤组织中表达呈正相关关系(r=0.516,P<0.05).结论 VEGF与AQP4是参与形成肿瘤周围水肿的重要分子生物学因素,且两者可能存在某种协同作用.     

不同病理级别脑胶质瘤的肿瘤干细胞体外分离培养和增殖分化

目的探讨不同病理级别的人脑胶质瘤的肿瘤干细胞在体外分离培养和增殖分化的情况。方法通过对不同病理级别手术切除的胶质瘤新鲜标本在体外无血清培养基中分离培养和体外增殖分化进行观察研究和分组比较它们的增殖能力。结果在无血清培养基中,Ⅰ级胶质瘤的肿瘤干细胞不能形成克隆球;Ⅱ级胶质瘤的肿瘤干细胞可形成中等克隆球;只有Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞才能形成大型克隆球。在血清培养基中,Ⅰ级胶质瘤干细胞增殖缓慢、分化较晚;Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞增殖迅速、快速进入分化期;Ⅱ级胶质瘤肿瘤干细胞增殖分化介于两者之间。分组比较,发现在不同培养基中和不同病理级别胶质瘤干细胞各组之间在增殖和分化能力上均存在统计学差异(P<0.01)。结论胶质瘤的肿瘤干细胞在生物特性上存在多样性。Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞易于分离培养,增殖分化能力较强;Ⅰ级胶质瘤的肿瘤干细胞增殖分化缓慢,有助于动态观察脑肿瘤干细胞自我更新和增殖分化的过程,是一个较好的观察研究对象。     

Scanning electron microscopy of malignant gliomas. A comparative study of glioma cells in smear preparations and in tissue culture

Summary Smear preparations from 15 malignant gliomas, 2 metastatic carcinomas and from normal brain were examined by scanning electron microscopy. Tissue culture preparations from malignant gliomas were also studied. In the better differentiated areas of gliomas, the cells in smears were stellate with multiple long interweaving processes 0.25–1.3 m in diameter which could be distinguished from myelinated nerve fibres (1.3–5 m) and from fibrin (0.08–0.3 m) by their thickness and arrangement in the tissue. The relationship of glial processes to blood vessels within the tumour was well demonstrated in smears. Metastatic carcinoma cells lacked the processes seen in glioma cell smears and did not show the same relationship to blood vessels. The more anaplastic glioma cells had fewer processes and ovoid cell bodies covered with surface ruffles and microvilli similar to the cell membrane projections in the nuclear regions of glioma cells in culture. The relationship of the surface morphology of glioma cells in smears to the known invasive nature of these tumours is discussed.     

神经电生理监测技术在功能区胶质瘤术中的应用

目的 探讨神经电生理监测在功能区胶质瘤术中的应用价值。方法 对2012年1月至2014年12月术中行神经电生理监测的738例功能区胶质瘤的临床资料进行回顾性分析。结果 608例在全麻下行术中神经电生理监测,130例在唤醒麻醉下行运动和或语言功能定位。所有病例术后近期运动障碍发生率为11.7%,远期为3.8%;语言区胶质瘤术后近期的失语率为28.8%,远期为4.1%。术中MRI辅助下进行电生理监测347例,全切率为89.3%。全麻下3.6%的患者术中出现癫痫大发作;唤醒麻醉下2.3%的患者出现一侧肢体或嘴角抽搐,仅1例（0.8%）出现癫痫大发作。结论 根据胶质瘤所在的部位选择适当的术中神经电生理监测技术,有助于最大程度切除肿瘤的同时,保护患者的功能皮层和皮层下重要功能通路,降低致残率,提高远期生活质量。     

Cyclin D1 expression in gliomas

Cyclin D1 (cycD1) expression was defined immunohistochemically using monoclonal antibody DCS-6 and polyclonal antiserum H-295 in 50 glioma biopsies. The number of positive nuclei was higher for H-295 than for DCS-6, with a ratio of 3:1. The labelling index (LI) was compared to the grade of histological malignancy and to Ki-67 MIB-1 LI. The LI for cycD1 increased with histological malignancy, in parallel with the increase in MIB-1 LI. In most tumours, the maximum LI for cycD1 and MIB-1 were found in the same areas. The mean MIB-1 LI: mean cycD1 LI ratio does not vary in the three grades of astrocytic tumours. However, in this study the correlation between the two LIs was not statistically significant. Staining for cycD1 antigen does not necessarily imply that the gene is overexpressed since other molecular mechanisms can also be responsible for cell cycle deregulation. In invasive areas, the cycD1 LI is frequently higher than in solid tumour, either because more tumour cells are positive or because reactive astrocytes and activated microglia express cycD1. The relative contribution of neoplastic and reactive cells remains to be defined. Received: 23 May 1997 / Revised: 21 July 1997 / Accepted: 20 August 1997     

术前预康复在脑胶质瘤手术病人中的应用

目的 探讨术前预康复在脑胶质瘤手术病人中的应用价值。方法 回顾性分析2015年12月至2018年12月手术治疗的80例脑胶质瘤的临床资料。40例进行常规术前准备（对照组）,40例在对照组基础上在进行术前预康复（观察组）。干预前、术前1 d及术后4周,使用Fugl-Meyer运动功能量表（FMA）评分评价运动能力,改良Barthel指数量表（MBI）评分评价日常生活能力,医院焦虑抑郁量表（HADS）评分评价心理状态,中国头颈癌生命质量测定量表（QLQCP-HN）评分评价生命质量。结果 术前1 d、术后4周,观察组FMA、MBI、QLQCP-HN评分均明显高于对照组（P<0.05）,而HADS评分明显低于对照组（P<0.05）。结论 术前预康复可改善脑胶质瘤病人术后运动能力、心理状态,提高日常生活能力及生命质量     

丘脑胶质瘤的临床特征与手术治疗的分析

目的研究丘脑胶质瘤的临床特征、诊疗方法和手术技巧。方法回顾性分析我院经手术治疗的39例丘脑胶质瘤患者的临床表现、肿瘤影像学特点和手术治疗效果。结果丘脑不同类型胶质瘤影像学表现、全切率不尽相同。病程7d～3．5年，平均3．1月。肿瘤颅内压增高症状最为常见（90％），轻偏瘫56％，不自主运动10％，智力减退或精神症状30％；全组患者肿瘤手术全切率为53％，次全切率为29％，部分切除率为18％；出院时好转、稳定者占79％，恶化及出现新体征者占21％；手术死亡率为0。结论MRI是目前诊断丘脑肿瘤的最好检查方法。肿瘤的病理分类与临床症状、影像特征及手术效果均有关。