多形性胶质母细胞瘤预后因素分析

目的 探讨多形性胶质母细胞瘤(GBM)预后相关的临床因素.方法 回顾性分析160例随访资料完整的GBM,采用Kaplan-Meier法计算生存率及进行单因素分析,Log-rank法进行生存率显著性检验,Cox比例风险回归模型作多因素分析.结果 本组病人术后1年、2年、5年生存率分别是65.3％、34.1％、9.1％.单因素分析显示年龄、肿瘤切除程度、治疗方式是影响GBM生存率的因素.多因素分析显示年龄、肿瘤部位、肿瘤切除程度及治疗方式是与GBM预后相关的独立因素.结论 年龄、肿瘤部位、肿瘤切除程度和治疗方式是影响GBM预后的主要因素,积极的手术治疗和规范化、个体化的综合治疗可有效提高GBM病人的生存率.     

High expression of RFX4 is associated with tumor progression and poor prognosis in patients with glioblastoma

Abstract

Aim: Glioma stem cells (GSCs) have been shown to contribute to tumor development and recurrence, therapeutic resistance, and cellular heterogeneity of glioblastoma multiforme (GBM). Recently, it has been reported that GSCs lose their self-renewal ability and tumorigenic potential upon differentiation. In this study, we identified Regulatory Factor X4 (RFX4) gene to regulate GSCs’ survival and self-renewal activity in the GBM patients samples.

Materials and methods: We utilized public datasets from the Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), Ivy Glioblastoma Atlas Project, and The Human Protein Atlas to screen candidate genes which are associated with the development of GBM and poor patients survival. Small hairpin RNA (shRNA) lentivirus was applied to knockdown RFX4 gene in GSCs.

Results: We found that RFX4 mRNA expression among the RFX family was particularly reduced during GSC differentiation. RT-qPCR analysis revealed significant downregulation of RFX4 and stem cell markers (CD15 and CD133) mRNA expressions in primary human GBM-derived GSCs cultured under serum condition. Consistently, GSCs showed significantly elevated RFX4 mRNA expression levels compared to normal astrocytes, NHA, whereas glioma cells did not. Furthermore, analysis of the TCGA data set revealed that RFX4 is highly expressed in GBM, and contributes to the lowering of patient survival. Depletion of RFX4 using shRNA lentivirus in patient GBM-derived GSCs decreased neurosphere formation and cell viability.

Conclusion: These results suggest that RFX4 is a potential risk factor for maintaining the stemness of GSCs and making glioma more malignant, and thus, could be a promising target of GBM treatment.     

Time-dependent risk factors for epileptic seizures in glioblastoma patients: A retrospective analysis of 520 cases

Objective

Epilepsy is a common comorbidity of glioblastoma. Seizures may occur in various phases of the disease. We aimed to assess potential risk factors for seizures in accordance with the point in time at which they occurred.

Methods

We retrospectively analyzed medical files of adult patients with de novo glioblastoma treated at our institution between January 2006 and January 2020. We categorized seizures as preoperative seizures (POS), early postoperative seizures (EPS; before initiation of radio[chemo]therapy [RCT]), seizures during radiotherapy (SDR; during or <30 days after RCT), and posttherapeutic seizures (PTS; ≥30 days after completion of RCT). We addressed associations between patients' characteristics and their seizures.

Results

In the final cohort (N = 520), 292 patients experienced seizures. POS, EPS, SDR, and/or PTS occurred in 29.6% (154/520), 6.0% (31/520), 13.8% (70/509), and 36.1% (152/421) of patients, respectively. POS occurred more frequently in patients with higher Karnofsky Performance Scale scores (odds ratio [OR] = 3.27, p = .001) and tumor location in the temporal lobe (OR = 1.51, p = .034). None of the parameters we analyzed was related to the occurrence of EPS. SDR were independently associated with tumor location (parietal lobe, OR = 1.86, p = .027) and POS, but not EPS, and were independent of RCT. PTS were independently associated with tumor progression (OR = 2.32, p < .001) and with occurrence of SDR (OR = 3.36, p < .001), and negatively correlated with temporal lobe location (OR = .58, p < .014). In patients with tumors exclusively located in the temporal lobe, complete tumor resection was associated with a decreased risk of postoperative seizures.

Significance

Seizures in glioblastoma patients have various, time-dependent risk factors. Temporal lobe localization was a risk factor for preoperative seizures; surgery may have had a protective effect in these patients. RCT did not have dose-dependent pro- or anticonvulsive effects. PTS were associated with tumor progression.     

PKA and PKC activation induces opposite glial fibrillary acidic protein (GFAP) expression and morphology changes in a glioblastoma multiform cell line of clonal origin

Possible differentiation mechanisms were investigated in a glioblastoma multiform cell line (GL15) presenting an undifferentiated phenotype with weak glial fibrillary acidic protein (GFAP) and strong vimentin (VIM) expression. Serum-free conditions induced time-dependent increases of GFAP-mRNA and GFAP protein levels, associated with a process-bearing astrocytic morphology. Activation of protein kinase C (PKC) by tumor promoter phorbol 12-myrystate 13-acetate (PMA) induced a rapid morphological differentiation and a decrease in GFAP mRNA, whereas the GFAP level remained unchanged. Such parameters were shown to characterize a physiological differentiation stage in astroglial cultures. Treatment of process-bearing GL15 cells with dibutyryl cyclic AMP (dbcAMP), a protein kinase A (PKA) activator, induced a timedependent decrease in the GFAP mRNA and GFAP protein levels and reverted morphological changes induced by serum-free conditions. Neither PMA nor dbcAMP influenced the VIM mRNA expression. In GL15 cells, PKC and PKA activation have opposite effects. Understanding the role of these kinases in malignant transformation and in the in vitro differentiation process is of both basic and clinical interest. © 1995 Wiley-Liss, Inc.     

rCBV变化与VEGF表达对胶质母细胞瘤术后放疗病人生存期的预测研究

目的 探讨相对脑血容量(relative cerebral blood volume,rCBV)变化和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达对胶质母细胞瘤全切手术后辅助放射治疗病人的无进展生存期(PFS)和总生存期(OS)的预测作用.方法 回顾性分析16例外科全切术后辅助放射治疗的胶质母细胞瘤病人的临床资料.在放射治疗前和放射治疗中(累计放射剂量为30 Gy)各进行1次灌注成像检查,计算rCBV值.免疫组化检测VEGF表达.采用Kaplan-Meier生存曲线分析PFS和OS.结果 不同年龄、VEGF表达和rCBV变化的病人,其在PFS的差异具有统计学意义(均P＜0.05).仅有VEGF表达不同的病人在OS的差异具有统计学意义(P＜0.05).结论 VEGF表达和rCBV变化可以作为判定胶质母细胞瘤PFS的预测物.同时,VEGF表达可以作为判定胶质母细胞瘤OS的预测物.     

Impact of molecular and clinical variables on survival outcome with immunotherapy for glioblastoma patients: A systematic review and meta‐analysis

BackgroundGiven that only a subset of patients with glioblastoma multiforme (GBM) responds to immuno‐oncology, this study aimed to assess the impact of multiple factors on GBM immunotherapy prognosis and investigate the potential predictors.MethodsA quantitative meta‐analysis was conducted using the random‐effects model. Several potential factors were also reviewed qualitatively.ResultsA total of 39 clinical trials were included after screening 1317 papers. Patients with O6‐methylguanine‐DNA methyltransferase (MGMT) promoter methylation [hazard ratio (HR) for overall survival (OS) = 2.30, p < 0.0001; HR for progression‐free survival (PFS) = 2.10, p < 0.0001], gross total resection (HR for OS = 0.70, p = 0.02; HR for PFS = 0.56, p = 0.004), and no baseline steroid use (HR for OS = 0.52, p = 0.0002; HR for PFS = 0.61, p = 0.02) had a relatively significant favorable OS and PFS following immunotherapy. Patients with a Karnofsky Performance Status score < 80 (HR = 1.73, p = 0.0007) and undergoing two prior relapses (HR = 2.08, p = 0.003) were associated with worse OS. Age, gender, tumor programmed death‐ligand 1 expression, and history of chemotherapy were not associated with survival outcomes. Notably, immunotherapy significantly improved the OS among patients undergoing two prior recurrences (HR = 0.40, p = 0.008) but not among patients in any other subgroups, as opposed to non‐immunotherapy.ConclusionSeveral factors were associated with prognostic outcomes of GBM patients receiving immunotherapy; multiple recurrences might be a candidate predictor. More marker‐driven prospective studies are warranted.     

脑脊液14-3-3蛋白分析在评价多发性硬化轴索损害中的临床价值

目的 评价脑脊液14-3-3蛋白与多发性硬化（MS）患者轴索损害之间的关系。方法 从47例MS患者及36例对照中获取脑脊液标本，用免疫印迹法分析脑脊液14-3-3蛋白条带。结果 MS患者脑脊液14-3-3蛋白阳性率为24．5％，急性期起病的患者12例中7例阳性；阳性的患者其扩展的残疾功能状况量表评分均≥2．0分，且〉4．5分的患者阳性率更高。结论 免疫印迹法分析脑脊液14-3-3蛋白对判断MS患者急性期是否出现轴索损害及遗留神经功能缺损有一定的参考价值。     

Monitoring and tentative diagnosis of herpetic encephalitis by protein analysis of cerebrospinal fluid. Particular relevance of the assays of ferritin and S-100

The cerebrospinal fluid (CSF) of 9 patients with herpetic encephalitis was analyzed by particle counting immunoassay of ferritin, S-100, immunoglobulins, anti-herpes antibodies and immune complexes and by electrophoresis for the detection of oligoclonal bands. The main conclusions are: first, the simultaneous increase of both ferritin and S-100 in the presence of symptoms of encephalitis suggests strongly the infection is herpetic; second, high and increasing levels of S-100, probably related to the extent of the necrotic process, indicate a poor prognosis. In addition, 8-14 days after onset, locally produced anti-herpes antibodies were detectable, the IgG index increased and oligoclonal bands became visible.     

Apparent diffusion coefficient histogram analysis for prediction of prognosis in glioblastoma

Background

To investigate the potential to predict prognosis of glioblastoma (GBM) patients by analysis of the broader and lower values in the lower distribution of apparent diffusion coefficient (ADC_L) (B&L-ADC_L) values in the ADC histogram.

Kinetics and glial fibrillary acidic (GFA) protein production in a transplantable human giant cell glioblastoma (D-212 MG) of near haploid karyotype maintained in an organ culture system. An immunohistochemistry study

A transplantable subcutaneous tumour (designated D-212 MG), sequentially passaged in athymic nude mice and originally derived from a human giant cell glioblastoma, was maintained in an organ culture (matrix) system and studied immunohistochemically after in vitro pulse-labelling with bromodeoxyuridine (BrdU) and for the presence of glial fibrillary acidic (GFA) protein, after 1, 2 and 3 weeks in culture. The histological characteristics of the tumour, showing two cell populations of giant multinucleated cells and small cells, were preserved in the explants. An increased percentage of multinucleated giant cells was found after 3 weeks in vitro. A small but constant fraction (4-6%) of these cells continued to synthesize DNA. The labelling index of the small cells was somewhat higher, but decreased slightly although significantly over the 3-week period in vitro (from approximately 10.5 to 8%). The percentage of small cells that were positive for GFA protein was in the region of 75% and that of the giant multinucleated cells was in the region of 45%; it did not change significantly during the 3 weeks in vitro. The in vitro results confirm the astrocytic nature of both the small cells and the giant multinucleated cells in this tumour, the capacity of both cell populations to synthesize DNA in culture and to demonstrate invasiveness, and suggest the possibility that some of the giant multinucleated cells may have originated from the conversion of a number of small tumour cells.     

CSF tau protein is a useful marker for effective treatment of superficial siderosis of the central nervous system: Two case reports

We report two cases of superficial siderosis (SS) of the central nervous system (CNS), which is caused by chronic haemorrhaging into the subarachnoid space with haemosiderin deposition in the superficial portion of the CNS. Patient 1 had fluid collection in the spinal canal, which was reported as the source of the chronic bleeding. Patient 2 was bleeding from thickened dura at the level of the sacral vertebrae. Both of the patients had xanthochromic cerebrospinal fluid. We surgically repaired the sources of bleeding. Subsequently the cerebrospinal fluid (CSF) cleared and their symptoms were not aggravated for about 1 year. We measured several CSF markers of SS before and after surgery. Total tau protein (CSF-t-tau), phosphorylated tau protein (CSF-p-tau), iron (CSF-iron) and ferritin (CSF-ferritin) in the CSF were highly elevated at diagnosis. After surgery, the levels of CSF-t-tau and CSF-p-tau were markedly reduced while CSF-iron and CSF-ferritin had not decreased. It is suggested that CSF-t-tau and CSF-p-tau reflected the neural damage in SS and were useful to evaluate the effectiveness of SS therapies.     

Clinical analysis for an unusual large cystic meningioma: case report and review of the literature

The authors report the case of a 17-year-old boy with an unusual large cystic meningioma (Nauta type II) in the right hemisphere. The imaging appearances of this patient were very unusual. The shape of the huge cyst was crescentic and similar to subdural hematoma. It lay between the dura and the solid tumor parts. In addition there was a small intracystic nodule attached to the cyst wall. The patient underwent a right hemisphere craniotomy. At surgery it was found that the cyst contained a large amount of xanthochromic fluid and some semitransparent serumlike sediment. The intracystic nodule was proved to be necrotic substance without tissue and cell structure. Histological examination displayed an anaplastic meningioma, of which the cyst wall also consisted of meningioma tissue. To the best of the authors' knowledge, such an unusual case of cystic meningioma has not been reported. The authors review the literature with reference to intratumoral cyst associated with meningiomas, analyze the unusual imaging appearances of this patient, and explore the mechanism of cyst formation. The mechanism of cyst formation associated with meningiomas is not perfectly understood. Intratumoral cyst formation may be attributed to microcystic degeneration, ischemic necrosis, intratumoral hemorrhage, transudation and secretory changes within the tumor.     

A novel approach for assessing hypoperfusion in stroke using spatial independent component analysis of resting‐state fMRI

Individualized treatment of acute stroke depends on the timely detection of ischemia and potentially salvageable tissue in the brain. Using functional MRI (fMRI), it is possible to characterize cerebral blood flow from blood‐oxygen‐level‐dependent (BOLD) signals without the administration of exogenous contrast agents. In this study, we applied spatial independent component analysis to resting‐state fMRI data of 37 stroke patients scanned within 24 hr of symptom onset, 17 of whom received follow‐up scans the next day. Our analysis revealed “Hypoperfusion spatially‐Independent Components” (HICs) whose spatial patterns of BOLD signal resembled regions of delayed perfusion depicted by dynamic susceptibility contrast MRI. These HICs were detected even in the presence of excessive patient motion, and disappeared following successful tissue reperfusion. The unique spatial and temporal features of HICs allowed them to be distinguished with high accuracy from other components in a user‐independent manner (area under the curve = 0.93, balanced accuracy = 0.90, sensitivity = 1.00, and specificity = 0.85). Our study therefore presents a new, noninvasive method for assessing blood flow in acute stroke that minimizes interpretative subjectivity and is robust to severe patient motion.     

改良额底入路切除鞍结节脑膜瘤31例

目的探讨改良额底入路切除鞍结节脑膜瘤的临床效果。方法回顾性分析本人自2009年12月至2013年12月来采取不损伤颞肌,不选传统额底入路的"关键孔"点,而选取"美容孔"点,骨瓣大小和位置选择灵活的改良额底入路切除鞍结节脑膜瘤31例的临床资料。结果手术后4例病人癫痫未再发作,无垂体功能低下病例,无尿崩病例,无额纹消失病例,无颞肌萎缩病例。有5例术侧嗅神经断裂。术前有48只眼视力下降,术后有46只眼视力明显改善,有1只眼视力无明显变化,1只眼视力下降。simpsonⅠ级切除25例中,有1例于术后2年复发。simpsonⅡ级切除5例中,有1例于术后1.5年时复发。simpsonⅢ级切除1例,随访2.5年残余肿瘤无明显变化。结论改良额底入路切除鞍结节脑膜瘤具有灵活、安全、暴露充分、切除彻底、损伤轻等优点。     

Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Isolated brain capillaries are essential for analyzing the changes of protein expressions at the blood–brain barrier (BBB) under pathological conditions. The standard brain capillary isolation methods require the use of at least five mouse brains in order to obtain a sufficient amount and purity of brain capillaries. The purpose of this study was to establish a brain capillary isolation method from a single mouse brain for protein expression analysis. We successfully isolated brain capillaries from a single frozen mouse brain by using a bead homogenizer in the brain homogenization step and combination of cell strainers and glass beads in the purification step. Western blot and proteomic analysis showed that proteins expressed at the BBB in mouse brain capillaries isolated by the developed method were more enriched than those isolated from a pool of five mouse brains by the standard method. By using the developed method, we further verified the changes in expression of BBB proteins in Glut1-deficient mouse. The developed method is useful for the analysis of various mice models with low numbers and enables us to understand, in more detail, the physiology and pathology of BBB.     

Comparison of a non-competitive two-site RIA and an inhibition RIA for the detection of myelin basic protein in cerebrospinal fluid

Two radioimmunoassay (RIA) produces were used to measure human myelin basic protein (HBP) in cerebrospinal fluid (CSF): (1) an inhibition RIA, with the use of TNP-conjugated anti-BP IgG, ¹²⁵I-labelled HBP, and anti-TNP-coated polystyene beads, and (2) a non-competitive two-site RIA, with the use of Sepharose-coupled anti-BP antibodies and ¹²⁵I-labeled anti-BP IgG.The two-site RIA detects less HBP in CSF than the inhibition RIA, partly due to the presence of HBP fragments in CSF that are detected by the inhibition asasy, but less by the two-site RIA. The correlation was improved when in the two-site RIA Sepharose-coupled anti-BP antibodies were changed. Because certain substrates (such as autoantibodies to HBP) may give false-positive results in the competitive RIA but not in the two-site RIA, we conclude that a combination of the (more sensitive) inhibition RIA with the (more specific) two-site assay provides a more reliable HBP assay than either assay alone.     

Detailed knowledge of cellular expression of G protein‐coupled receptors in the human enteric nervous system is essential for understanding their diverse actions

G protein‐coupled receptors (GPCRs) comprise a large and diverse superfamily of transmembrane receptors that mediate the functions of an extraordinarily large number of extracellular ligands. They control many major physiological processes and are involved in diverse pathological processes, including gastrointestinal diseases. G protein‐coupled receptors are one of the most targeted classes in pharmaceutical drug research. At present, much of our knowledge concerning the expression, distribution and function of GPCRs in the gut has been gleaned from studies performed in experimental models. Data obtained in the human digestive tract, especially in the enteric nervous system, are sparse and incomplete, although enteric neurons have a key position in almost all physiological and pathophysiological processes in the gut. Knowledge of cellular distribution of GPCRs, of regional differences in GPCR expression, and of altered GPCR expression during pathophysiological conditions in the human gut, will lead to a better understanding of GPCR activity, but will also contribute to the development of new drugs. In the current issue of the Journal, Harrington et al. describe the presence and cellular localization of muscarinic receptors in the human colon. Morphologically, orientated studies on the cellular expression of GPCRs in the human gut have to be encouraged, because these studies will yield data that are of therapeutic relevance.     

The G79A polymorphism of protein Z gene is an independent risk factor for cerebral venous thrombosis

Background and purpose   Protein Z (PZ), a vitamin Kdependent protein, plays a role in inhibiting coagulation. Its plasma level or PZ gene polymorphisms have been discussed as risk factors for stroke with conflicting results reported between various studies. Only one of these polymorphisms was studied in a cohort of patients suffering from cerebral venous thrombosis (CVT). Methods   We performed a retrospective genetic study comparing 100 healthy controls to 54 patients referred to our hemostasis unit after CVT occurrence. We compared the distribution of three PZ gene polymorphisms that may influence PZ plasma levels: A-13G in the promoter and G79A in intron F were tested using previously described techniques, and we developed a technique to evaluate the G-103A in intron A. Results   The G79A polymorphism was significantly more frequent in patients than in controls (p = 0.012): the presence of at least one A allele led to an odds ratio of 2.57 with a 95 % confidence interval of 1.23–5.34. The A-13G polymorphism also showed a nonsignificant trend towards a higher prevalence in patients. Conclusion   The G79A polymorphism of the PZ gene was shown to be a new independent risk factor for cerebral venous thrombosis. Nevertheless, these results have to be confirmed by a prospective study including plasma PZ evaluation.