BACKGROUND:Low-grade gliomas (LGG) are a variety of brain tumors that show different clinical outcomes. The methylation of the GSTM5 gene has been noted in the development of LGG, however, its prognostic importance remains uncertain. OBJECTIVE:The objective of this study was to examine the correlation between GSTM5 DNA methylation and clinical outcomes in individuals diagnosed with LGG. METHODS:Analysis of GSTM5 methylation levels in LGG samples was conducted using data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. The overall survival based on GSTM5 methylation status was evaluated using Kaplan-Meier curves. The DNA methylation heatmap for particular CpG sites in the GSTM5 gene was visualized using the "pheatmap" R package. RESULTS:The study analyzed that LGG tumors had higher levels of GSTM5 methylation than normal tissues. There was an inverse relationship discovered between GSTM5 expression and methylation. LGG patients with hypermethylation of GSTM5 promoter experienced a positive outcome. Age, grade, and GSTM5 methylation were determined as independent prognostic factors in LGG through both univariate and multivariate Cox regression analyses. CONCLUSION:Methylation of GSTM5 DNA, specifically at certain CpG sites, is linked to a positive outlook in patients with LGG. Utilizing the "pheatmap" R package to visualize GSTM5 methylation patterns offers important information for identifying prognostic markers and therapeutic targets in low-grade gliomas.

Phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP) is a new-found tumor suppressor in a variety of tumors. While, it is still unknown about its role in glioma. In this study, we found that LHPP is abnormally decreasing or absent in glioblastoma, and the low expression of LHPP is associated with poor median survival in glioma patients. Functional assay revealed that LHPP-overexpression significantly inhibited U87MG and U118MG growth in vitro and in vivo. As to the mechanism, mass-spectrometric analysis indicated that the LHPP interacting proteins were mainly enriched in regulation of energy metabolism, including Carbon metabolism, Oxidative phosphorylation, and Glycolysis. Seahorse assay and metabolites detection confirmed that LHPP-overexpression obviously impeded glycolysis and respiration in U87MG and U118MG cells. For the further study, western blot assay showed that the protein level of PKM2 at dimeric, tetrameric, and total protein, were all decreased significantly, and its enzymatic activity was decreased as well. ChIP and RNAseq integrated analysis indicated that the decreased protein level of PKM2 was independent of PKM2 transcription, and LHPP did not reprogram transcription level of metabolic genome. Co-IP and immunofluorescence assay manifested that LHPP interacted with PKM2, and this interaction interfered the protein stability, then induced ubiquitin-mediated degradation of PKM2. Rescue assay confirmed that restoring the expression of PKM2 effectively reversed the restrained energy metabolism and the inhibited cancer cell growth caused by LHPP-overexpression in U87MG and U118MG cells. Taking together, we demonstrated that LHPP impedes the glycolysis and respiration during energy metabolic process via inducing ubiquitin-mediated degradation of PKM2, thus inhibits the growth of glioblastoma.

Background P7C3 is a neurogenic compound that exhibits neuroprotective properties in neural cells. However, its target proteins and effects in glioma are unknown. Methods The candidate P7C3 target proteins were analyzed using a human protein microarray containing 23136 human proteins. A streptavidin agarose affinity assay was used to verify the direct interaction between P7C3 and phosphoglycerate kinase 1 (PGK1). Mass spectrometry was used to identify the binding sites of PGK1 for P7C3 binding. Seahorse XF96 extracellular flux analyzer was used to measure the cell oxygen consumption rate and extracellular acidification rate. Glycolytic metabolites were measured using the related kits. Protein level was detected by western blotting and immunohistochemical staining. Autophagy was analyzed using a transmission electron microscope and western blotting. The malignancy of tumor progression in vitro and in vivo was analyzed based on cell viability, apoptosis and proliferation, migration and invasion, and xenograft model. Glial cells were marked by antibodies via immunohistochemical staining. Results The human protein microarray identified 577 candidate P7C3 target proteins. The global profile of P7C3 target proteins indicated that P7C3 regulates glycolysis. Metabolic experiments confirmed that P7C3 regulates aerobic glycolysis in glioma cells. The underlying mechanism of P7C3 was found to be direct targeting PGK1 at lysine residues and asparagine residues, and the specific P7C3-PGK1 interaction led to decreased protein level and total intracellular kinase activity of PGK1. The Cancer Genome Atlas and Chinese Glioma Genome Atlas databases indicated that the mRNA level of PGK1 is significantly increased in high-grade glioma, and the abnormally high mRNA level of PGK1 is associated with a poor prognosis in patients with glioma, suggesting that PGK1 is a promising target for glioma therapy. The inhibition of PGK1 and the subsequent suppression of aerobic glycolysis caused by P7C3 inhibited the malignant growth of glioma in vitro and in vivo. Furthermore, P7C3 did not damage normal glial cells under concentration, which exhibit an inhibitory effect on gliomas. Conclusions This study revealed that P7C3 suppresses glioma by regulating aerobic glycolysis via directly targeting PGK1. Furthermore, we identified the P7C3 target proteins for the first time which is expected to provide scientific clues for future studies.

Atypical extraventricular neurocytoma (EVN) is a rare condition characterized by diffuse tumor cell hyperplasia, increased neovascularization, increased necrosis, and aggressive characteristics. A case of a 25-year old man who presented with atypical EVN in his left parietal - occipital flaps is reported. Magnetic resonance imaging (MRI) revealed a well-defined globular mass with heterogeneous signals in the left parietal lobe, and mild perilesional edema. After left parietal craniotomy and tumor excision, pathologic examination of the resected tissue revealed that the lesion was localized mainly in the white matter and imbued with tumor cells possessing round hyperchromatic nuclei with perinuclear halos and increased microvascular proliferation. The patient underwent radiotherapy at 21st postoperative day. Over the past 26 months, the patient has been regularly followed up, and so far no neurologic deficits have been observed. The latest MRI showed that the tumor bed was stable with slight peritumoral edema. The results of clinical, histopathological and immunohistochemical examinations indicate that atypical EVN is a rare neoplasm with unique radiographic and pathologic characteristics. It possesses more aggressive properties than typical EVN.

The aim of this study was to investigate the effect of three-dimensional (3D) bio-printed constructs consisting of human umbilical-derived mesenchymal stem cells (HUMSCs) on cell viability, proliferation and differentiation in vitro. Functional 3D bio-printed microspheres consisting of HUMSCs were constructed using electrostatic inkjet technique. The parameters used for the synthesis of 3D bio-printed tissue constructs were first optimized. The viability, proliferation and differentiation of 3D cultured HUMSCs were assessed. The results of scanning electron microscopy (SEM) showed that isolated HUMSCs exhibited fibroblast-like spindle adherent growth. The optimized printing parameters were 6 kV voltage, 10 mL/h flow, 15 cm receiving height, and alginate: water ratio of 1:1 mixed at 37 °C. Compared with 2D cultured HUMSCs, the 3D cultured HUMSCs have better viability, proliferation and differentiation ability. The results obtained in this study indicate that 3D bio-printed tissue constructs promote HUMSC viability, proliferation, and neural differentiation in vitro.

NSI-566 is a stable, primary adherent neural stem cell line derived from a single human fetal spinal cord and expanded epigenetically with no genetic modification. This cell line is being tested in clinical trials in the U.S. for treatment of amyotrophic lateral sclerosis and spinal cord injury. In a single-site, phase I study, we evaluated the feasibility and safety of NSI-566 transplantation for the treatment of hemiparesis due to chronic motor stroke and determined the maximum tolerated dose for future trials. Three cohorts (n = 3 per cohort) were transplanted with one-time intracerebral injections of 1.2 x 10(7), 2.4 x 10(7), or 7.2 x 10(7) cells. Immunosuppression therapy with tacrolimus was maintained for 28 days. All subjects had sustained chronic motor strokes, verified by magnetic resonance imaging (MRI), initiated between 5 and 24 months prior to surgery with modified Rankin Scores [MRSs] of 2, 3, or 4 and Fugl-Meyer Motor Scores of 55 or less. At the 12-month visit, the mean Fugl-Meyer Motor Score (FMMS, total score of 100) for the nine participants showed 16 points of improvement (p = .0078), the mean MRS showed 0.8 points of improvement (p = .031), and the mean National Institutes of Health Stroke Scale showed 3.1 points of improvement (p = .020). For six participants who were followed up for 24 months, these mean changes remained stable. The treatment was well tolerated at all doses. Longitudinal MRI studies showed evidence indicating cavity-filling by new neural tissue formation in all nine patients. Although this was a small, one-arm study of feasibility, the results are encouraging to warrant further studies. Stem Cells Translational Medicine 2019;8:999-1007

目的 观察大型听神经瘤显微手术治疗的效果.方法 选取我院2010年4月~2017年1月72例大型听神经瘤患者,所有患者均经枕下乙状窦后入路行听神经瘤显微手术切除,观察患者的临床治疗效果与术后不良事件的发生情况.结果 肿瘤全切除患者共有62例,全切率为86.11%,次全切7例,占9.72%;大部切除3例,占4.16%.术中有64例患者面神经解剖保留,占88.89%;有50例患者术后1周面神经功能保留,占69.44%.术后肿瘤部位血肿1例,占1.38%;无术后死亡患者.结论 利用枕下乙状窦后入路显微手术方法能够有效提升患者的治愈率,并有效降低术后并发症与不良反应,值得临床推广.

The limited degree of functional recovery is closely associated with the condition of the periinfarct cortex after ischemic stroke. The model of oxygen-glucose deprivation in cultured neurons could partly simulate this condition. Proper dendritic morphology is crucial for the correct wiring of neuronal function. Hence, the question of how to facilitate the plasticity of neural dendrites is of great significance. DL-3-n-butylphthalide is an efficient medication for ischemic stroke. In this study, in addition to having neuroprotective effects, DL-3-n-butylphthalide could increase the number of primary and secondary dendrites and of dendritic tips as confirmed by Sholl analysis. This study further demonstrated that DL-NBP inactivates PI3K/AKT signaling to positively regulate dendritic branching.

目的研究总结骨髓间充质干细胞条件培养基对大鼠慢性脊髓损伤后胶质纤维酸性蛋白表达的影响.方法随机选取脊髓神经损伤研究实验室84只雄性Wistar大鼠,两组大鼠均为慢性脊髓损伤,依据大鼠的受伤情况并运用随机数字表法将其分为对照组(n=42)与观察组(n=42),对照组予以等渗盐水治疗,观察组予以BMSC-CM治疗.观察两组大鼠受伤后1、4、11、21、35、60、90d后的免疫组织化学检测结果,同时观察大鼠生物行为学变化与组织病理学变化.结果观察组大鼠的伤后恢复时间为(21.01±0.28)d,明显短于对照组(30.05±1.25)d,并且在11、21d的BBB评分与对照组相比差异较大;同时观察组大鼠在受伤第4,11,21天时的GFAP灰度值与对照组相比差异较大,具有统计学意义(P<0.05),在第21天至痊愈期间观察组大鼠的GFAP平均灰度值下降水平快于对照组.结论BMSC-CM能够有效的修复大鼠慢性脊髓损伤,并且能够降低胶质纤维酸性蛋白表达,对于损伤大鼠脊髓功能的恢复具有重要意义.

Aberrant activation of Wnt/β-catenin signaling leads to increased cell proliferation and survival and promotes the development of various human tumors, including glioma, one of the most common primary brain tumors. The treatment efficacy of many anticancer drugs remains limited or unsatisfactory and it is urgently necessary to develop effective and low-toxicity anticancer drugs or strategies, especially for glioma. Here, we report that diallyl trisulfide suppresses survival, migration, invasion and angiogenesis in glioma cells. These effects were associated with inhibition of the Wnt/β-catenin signaling cascade, which was accompanied by decreased expression of LRP6, TRIM29 and Pygo2. A dual-luciferase reporter assay confirmed that DATS treatment decreased TCF/LEF-mediated transcription. Finally, a nude mouse tumorigenicity model was used to examine the biological effect of diallyl trisulfide in vivo. Consistent with the previous results, diallyl trisulfide inhibited proliferation, invasion and angiogenesis in glioma cells by suppressing Wnt/β-catenin signaling.

Objective To evaluate the influence of SOCS3 conditional ablation in macrophage polarization and functional recovery after spinal cord injury in mice.Methods The Socs3fVfl Nes cre mice were used to obtain nervous-system-restricted SOCS3 deletion models as experimental group (n=40),and littermate Socs3fl as control group (n=40).Mice (n=35) in each group were underwent T10 spinal cord compressing injury (the left 5 were as sham-operated controls);on the third,7th and 14th d of injury,injured tissues from 15 mice of each group were harvested;immunofluorescence was performed to detect the changes of M2a and M2c phenotypic marker Arginase1 and M1 and M2b phenotypic marker CD86;on the first,third,7th and 14th d of injury,injured tissues from the left 20 mice from each group were harvested;fluorescent quantitative real time-PCR was used to detect the macrophage phenotypic markers (Arginasel,CD206,iNOS and CD32),SOCS3 and STAT3 gene expressions;the above data from the sham-operated controls were used as base data.BMS scale was used to evaluate the motor functions of posterior limbs and bodies on the first,third,7th and 14th d of injury.Results The behavioral assessments of the experimental group were superior on the third,7th and 14th d of injury as compared with those of control group (P＜0.05).Immunofluorescence showed that as compared with the controls,the Socs3fVfl Nes cre mice had a predominance in distribution and population of Arginasel-positive macrophages in spinal cords on the third,7th and 14th d of injury;in contrast,the control group showed superiority of CD86 expression on the 7th and 14th d,with significant differences (P＜0.05).Similarly,fluorescent quantitative real time-PCR analysis demonstrated that gene expression of A rginase I on the 7th d of injury,that of CD206 on the 7th and 14th d of injury,and that of STAT3 on the third,7th,and 14th d of injury were significantly increased,whereas that of CD32 on the 14th d of injury and SOCS3 on the first and third d of injury were significantly reduced in the experimental group,as compared with those in the control group (P＜0.05).Conclusion SOCS3 conditional knockout can promote the formation of M2 macrophage,as well as good functional recovery after spinal cord injury in mice.

Objective To discuss the effects of Protocadherin 11 X-linked protein (Pcdh11x) on formation of neuronal dendritic branching. Methods In this study, using gain-of-function and loss-of-function studies, we count the number of neural dendritic branching, and analyzed the data with SPSS 13.0 software by one way ANOVA. Results Compared with the control GFP vector, Pcdh11x-shRNA increased, whereas pCAG-Pcdh11x-GFP decreased the number of primary and secondary dendrites. The number of dendritic tips was reduced in neurons overexpressing pCAG-Pcdh11x-GFP, whereas it was increased when Pcdh11x activity was inhibited by overexpression of Pcdh11x-shRNA. The effects of Pcdh11x-shRNA and pCAG-Pcdh11x-GFP on dendritic branching were confirmed by Sholl analysis. Therefore pcdh11x acts as a negtive regulator of dendritic branching in cultured cortical neurons from embryonic day 16 mouse embryos. Conclusions Pcdh11x Negatively Regulates Dendritic Branching, including primary dendrites, secondary dendrites, dendritic tips and crossing number.

Allicin as a volatile oil content extracted from freshly garlic which is a complx includes a variety of allyl organosulfur components. One of the most biologically active ingredient of garlic. Allicin contents abundant of biological functions, lipid-lowering, anti-bacterial, alleviates inflammation, immune-enhancing, to prevent heart vascular disease and other physiological function. Compared with traditional chemical theropy, allicin is low residues in human and animals, non environmental pollution and has a broad resource. Recent study verify that allicin is able to obviously inhibite cancer cell proliferation, such as liver cancer, gastric cancer, colon carcinoma, lung cancer. Author briefly summarized recent research progress in the anti-tumor effect and mechanism of allicin. Key words: Allicin; Anticancer; Pharmacological action

Objective To compare the influence of three different puncture sites of in utero electroporation on the death rate of mice embryos, the thickness and the area of cortex, cell differentiation, cell proliferation, cell migration and cell apoptosis. Methods It groups on the bases of common puncture sites as follows: 1. electric shock only (control); 2. injection near the posterior fontanel, at about one millimeter (here, group A);3. injection along the antero-posterior axis into the cephalic ventricle (group B); and 4. injection near the midpoint between the anterior fontanel and posterior fontanel, at about three millimeters (group C). We compared the four groups using littermate mice, and repeated the experiment with 5 pregnant mice. To detect whether different injection positions affect cell differentiation, cell proliferation, cell migration and cell apoptosis, we stained cells with anti-Tbr1, anti-Tbr2, anti-Pax6 anti-Ctip2, anti- Caspase-3 and anti-Ki67. In differentiation, proliferation, and apoptosis assays, the cell number was analyzed with SPSS 13.0 software, using one way ANOVA. Results We found no statistical significant differences between the three puncture methods in the death rate of embryos, the thickness and the area of cortex, cell differentiation, cell proliferation, cell migration and cell apoptosis (P>0.05). Conclusion Three puncture sites used for in utero electroporation show no significantly different negative impacts during gene transfer into the embryonic mouse brain.

Objective To investigate the role of Wnt5a/Frizzled-2 signal in the process of nerve cell calcium overload after traumatic brain injury (TBI). Methods In vivo experience:adult Sprague-Dawley rats (n=96) were randomly divided into Sham group A (n=32), Pure injury group B (n=32) and RNAi inhibition group C (n=32). Moderate TBI modal was made in group B and C by Fenny method. RNAi was stereotactic hippocampal injected 48 hours before modal were made in group C to inhibit the expression of Frizzled-2. Rats were killed 24 h after injury, the levels of Frizzled-2 and Wnt5a in the injured side of hippocampal tissue were tested by western blotting method, the levels of calcium concentration were tested by immunofluorescent staining and laser confocal microscope. The data among groups was compared by single factor analysis of variance. Results In group A, the expression of Wnt5a/Frizzled-2 signal in the hippocampal tissue cells were stable. Compared with group A, the expression of Wnt5a and Frizzled-2 in the hippocampal tissue cells after TBI were increased by 2 times and 5 times (P<0.01) respectively in group B, and the level of calcium ion increased significantly (P<0.01). Compared with group B, with the inhibition of Frizzled-2, the expression of Wnt5a and Frizzled-2 in the hippocampal cells were decreased by 1 times and 3.55 times (P<0.01) respectively in group C, meanwhile the level of calcium ion were significantly decreased 1.5 times, close to the group A (P<0.01). Conclusion Under the physiological and pathological conditions, Wnt5a/Frizzled-2 signal plays an important role in the change of calcium concertration in the regulation of nerve cells. Our research suggests that the important factor related to the signal of Wnt5a, Frizzled-2, P-CamKII and RNAi which specific designed for Frizzled-2 RNAi could become potential therapeutic targets for further study in TBI.

Proper formation of neuronal dendritic branching is crucial for correct brain function. The number and distribution of receptive synaptic contacts are defined by the size and shape of dendritic arbors. Our previous research found that protocadherin 11 X-linked protein (Pcdh11x) is predominantly expressed in neurons and has an influence on dendritic branching. In this study, gain-of-function and loss-of-function experiments revealed that Pcdh11x acts as a negative regulator of dendritic branching in cultured cortical neurons derived from embryonic day 16 mice. Overexpression of wild-type Pcdh11x (Pcdh11x-GFP) reduced dendritic complexity, whereas knockdown of Pcdh11x increased dendritic branching. It was further demonstrated that Pcdh11x activates PI3K/AKT signaling to negatively regulate dendritic branching.

Although various ways to manipulate genes in vivo exist, in utero electroporation is a widely used technique, especially in the field of neural development due to its many advantages. In this study, we focused on direct comparison between three puncture sites during in utero electroporation on the death rate of embryos, the thickness and the area of cortex, cell differentiation, cell proliferation, cell migration and cell apoptosis. We found no statistical significant differences between the three puncture methods in the death rate of embryos, the thickness and the area of cortex, cell differentiation, cell proliferation, cell migration and cell apoptosis.

Protocadherin 11 X-linked (Pcdh11x) protein is a member of the cadherin superfamily with established roles in cell adhesion. Previous studies have shown the molecular biology and possible relevance of Pcdh11x with neurological disease in humans. However, little is known about the neurophysiological function of Pcdh11x in neural development. Here, we verified that Pcdh11x is primarily expressed in various brain areas including the cortex, hippocampus, and ventricular/subventricular zone (VZ/SVZ) at different embryonic stages. Furthermore, both in vitro and in vivo experiments showed that Pcdh11x decreased neural differentiation but increased the neural proliferation. These observations demonstrate a crucial function for Pcdh11x during the development of central nervous system.