An Electroconductive Hydrogel Scaffold with Injectability and Biodegradability to Manipulate Neural Stem Cells for Enhancing Spinal Cord Injury Repair.

Spinal cord injury (SCI) generally leads to long-term functional deficits and is difficult to repair spontaneously. Many biological scaffold materials and stem cell treatment strategies have been explored, but very little research focused on the method of combining exogenous neural stem cells (NSCs) with a biodegradable conductive hydrogel scaffold. Here, a NSC loaded conductive hydrogel scaffold (named ICH/NSCs) was assembled by amino-modified gelatin (NH2-Gelatin) and aniline tetramer grafted oxidized hyaluronic acid (AT-OHA). Desirably, the well-conducting ICH/NSCs can be simply injected into the target site of SCI for establishing a good electrical signal pathway of cells, and the proper degradation cycle facilitates new nerve growth. In vitro experiments indicated that the inherent electroactive microenvironment of the hydrogel could better manipulate the differentiation of NSCs into neurons and inhibit the formation of glial cells and scars. Collectively, the ICH/NSC scaffold has successfully stimulated the recovery of SCI and may provide a promising treatment strategy for SCI repair.

AUTOMATIC DETECTION AND GRADING OF DIABETIC MACULAR EDEMA BASED ON A DEEP NEURAL NETWORK.

PURPOSE: To solve the problem of automatic grading of macular edema in retinal images in a more stable and reliable way and reduce the workload of ophthalmologists, an automatic detection and grading method of diabetic macular edema based on a deep neural network is proposed. METHODS: The enhanced green channels of fundus images are input into the YOLO network for training and testing. Diabetic macular edema is graded according to the distance of the macula and hard exudate. We used multiscale feature fusion to form more comprehensive features on different grain images to improve the effect of hard exudate detection. We adopted K-means++ algorithm to cluster anchor box size and use loss of the original network to guide the regression of hard exudate bounding box and improve the regression accuracy of anchor boxes. We increased the diversity of samples for sample training by data augmentation, including cropping, flipping, and rotating of fundus images, so that each batch of training data can better represent the distribution of samples. RESULTS: The detection accuracy of the proposed method can reach 96% on the MESSIDOR data set. The detection rates of hard exudate with high, median, and low probability are 100%, 79.12%, and 60.40%, respectively. CONCLUSION: The proposed method exhibits a very good detection stability on healthy and diseased fundus images.

Hydrogel Composites with Different Dimensional Nanoparticles for Bone Regeneration.

The treatment of large segmental bone defects and complex types of fractures caused by trauma, inflammation, or tumor resection is still a challenge in the field of orthopedics. Various natural or synthetic biological materials used in clinical applications cannot fully replicate the structure and performance of raw bone. This highlights how to endow materials with multiple functions and biological properties, which is a problem that needs to be solved in practical applications. Hydrogels with outstanding biocompatibility, for their casting into any shape, size, or form, are suitable for different forms of bone defects. Therefore, they have been used in regenerative medicine more widely. In this review, versatile hydrogels are compounded with nanoparticles of different dimensions, and many desirable features of these materials in bone regeneration are introduced, including drug delivery, cell factor vehicle, cell scaffolds, which have potential in bone regeneration applications. The combination of hydrogels and nanoparticles of different dimensions encourages better filling of bone defect areas and has higher adaptability. This is due to the minimally invasive properties of the material and ability to match irregular defects. These biological characteristics make composite hydrogels with different dimensional nanoparticles become one of the most attractive options for bone regeneration materials.

Ca(Mg0.8Al0.2)(Si1.8Al0.2)O6:Ce3+,Tb3+ Phosphors: Structure Control, Density-Functional Theory Calculation, and Luminescence Property for pc-wLED Application.

A modified structure Ca(Mg0.8Al0.2)(Si1.8Al0.2)O6 (denoted as CMASO) from the evolution of CaMgSi2O6 (denoted as CMSO) codoped with Ce3+ and Tb3+ ions was designed successfully by solid reaction method for application in phosphor-converted white-light-emitting diode (pc-wLED). The Rietveld refinement of these two structures verified the changes derived from the replacement of some of the Mg2+ and Si4+ ions by Al3+ ions. The band gaps were calculated by density-functional theory (DFT) calculation method to verify the change of Al3+ ions replacing further, and the diffuse reflectance spectra (DRS) proved the veracity of the calculation result. The phosphors CMASO:Ce3+ showed blue emission excited by a wider excitation wavelength from 280 nm to 370 nm. The change of structure lead to the absorbable range broaden and the emission peak shifted to longer wavelength, compared with CMSO:Ce3+, although the amount of emitting center was the same. The reason for these phenomena was discussed in detail. The codoped phosphors CMASO:Ce3+,Tb3+ exhibited different emission colors from blue to green as the concentration of Tb3+ ions increased. Combined with commercial red phosphor CaAlSiN3:Eu2+ and ultraviolet LED (UV-LED) chips, the selected appropriate samples achieved white emission. The correlated color temperature (CCT) was 6137 K and the color rendering index (Ra) was 80.5, indicating that they could act as potential phosphors for possible applications in pc-wLED.

Topical Review: Causes of Refractive Error After Silicone-oil Removal Combined with Cataract Surgery.

SIGNIFICANCE: This review summarizes the main factors of refractive error after silicone oil removal combined with cataract surgery.The post-operative refractive results of silicone oil removal combined with cataract surgery are closely related to the patient's future vision quality. This report summarizes the factors that influence the difference between the actual post-operative refractive power and the pre-operatively predicted refractive power after silicone oil removal combined with cataract surgery, including axial length, anterior chamber depth, silicone oil, commonly used tools for measuring intraocular lens power, and intraocular lens power calculation formulas, among others. The aim of the report is to assist clinical and scientific research on the elimination of refractive error after silicone oil removal combined with cataract surgery.

Downregulated HDAC3 or up-regulated microRNA-296-5p alleviates diabetic retinopathy in a mouse model.

Objective: It has been demonstrated the efficacy of histone deacetylase 3 (HDAC3) in diabetes. Nevertheless, the function of HDAC3 in diabetic retinopathy (DR) remained largely obscure. Here, we investigated the HDAC3 effects in DR mice through the microRNA (miR)-296-5p/G protein subunit alpha i2 (GNAI2) axis. Methods: The mice diabetes model was established. HDAC3, GNAI2 and miR-296-5p levels in retina tissues of DR mice were evaluated. The weight, blood glucose, Evans blue leakage in DR mice, apoptosis of retinal ganglion cells, vascular endothelial growth factor (VEGF) and malondialdehyde (MDA) contents and superoxide dismutase (SOD) activity in DR mice were detected after miR-296-5p elevation or HDAC3 depletion. The relations among HDAC3, miR-296-5p and GNAI2 were validated. Results: HDAC3 and GNAI2 expressed at a high level while miR-296-5p expressed at a low level in retina tissues of DR mice. Restoring miR-296-5p or depleting HDAC3 reduced Evans blue leakage in DR mice, attenuated apoptosis of retinal ganglion cells, reduced VEGF and MDA, and enhanced SOD activity in serum and retinal tissues of DR mice. HDAC3 repressed miR-296-5p expression by binding to its promoter region, thereby enhancing GNAI2 expression. Conclusion: Depleting HDAC3 or restoring miR-296-5p suppresses apoptosis of retinal ganglion cells of DR mice via down-regulating GNAI2.

Lupeol induces autophagy and apoptosis with reduced cancer stem-like properties in retinoblastoma via phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin inhibition.

OBJECTIVES: To evaluate the anticancer effects of lupeol in retinoblastoma cells. METHODS: WERI-Rb-1 and Y-79 cell lines were used to evaluate the anticancer effect of lupeol. After lupeol treatment, the viability, proliferation, apoptosis, cancer stem-like properties, autophagy and in vivo tumour xenograft formation were detected. KEY FINDINGS: In this study, lupeol decreased cell viability in both WERI-Rb-1 and Y-79 cell lines. Lupeol could also inhibit proliferation and induce apoptosis of RB cells, with increased Bax level and decreased Ki67, survivin and Bcl-2 levels. Furthermore, lupeol could suppress the spheroid formation and stem-like properties of RB cells. Moreover, LC3 II/LC3 I ratio and the levels of Beclin1 and ATG7 were increased after lupeol treatment, indicating that lupeol could induce autophagy in RB cells. Next, the inhibitory effect of lupeol on the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway was observed. In tumour-bearing mice, lupeol suppressed tumour growth, and this might relate to its role in cell apoptosis, autophagy and stem-like properties. CONCLUSIONS: Lupeol suppressed proliferation and cancer stem-like properties, and promoted autophagy and apoptosis of RB cells by restraining the PI3K/AKT/mTOR pathway.

Joint optic disc and cup segmentation using feature fusion and attention.

Currently, glaucoma is one of the leading causes of irreversible vision loss. So far, glaucoma is incurable, but early treatment can stop the progression of the condition and slow down the speed and extent of vision loss. Early detection and treatment are crucial to prevent glaucoma from developing into blindness. It is an effective method for glaucoma diagnosis to measure Cup to Disc Ratio (CDR) by the segmentation of Optic Disc (OD) and Optic Cup (OC). Compared with OD segmentation, OC segmentation still faces difficulties in segmentation accuracy. In this paper, a deep learning architecture named FAU-Net (feature fusion and attention U-Net) is proposed for the joint segmentation of OD and OC. It is an improved architecture based on U-Net. By adding a feature fusion module in U-Net, information loss in feature extraction can be reduced. The channel and spatial attention mechanisms are combined to highlight the important features related to the segmentation task and suppress the expression of irrelevant regional features. Finally, a multi-label loss is used to generate the final joint segmentation of OD and OC. Experimental results show that the proposed FAU-Net outperforms the state-of-the-art segmentation of OD and OC on Drishti-GS1, REFUGE, RIM-ONE and ODIR datasets.

AuNCs-LHRHa nano-system for FL/CT dual-mode imaging and photothermal therapy of targeted prostate cancer.

As the most common cancer in men worldwide, prostate cancer has a serious impact on people's health. Until now, the development of a platform for integrating tumor targeting, imaging and an effective treatment for prostate cancer has remained challenging. Herein, a nano-system is designed to improve both diagnosis and treatment for prostate cancer. We successfully synthesized an AuNCs-LHRHa nano-system by combining PEI-modified gold nanoclusters (AuNCs) with LHRH analogues (LHRHa). Due to the good tunable optical properties and photothermal properties of AuNCs, the nano-system can not only achieve efficient fluorescence/computed tomography dual-mode imaging, but can also be used for photothermal therapy (PTT). After modifying the LHRHa antibody of a prostate tumor, AuNCs-LHRHa can be more effectively recognized by the gonadotropin-releasing hormone receptors (GnRH-R) on the membrane of RM-1 cells, enhancing the tumor cell uptake of the nano-system, improving the targeting accuracy and PTT therapy efficacy for prostate cancer. It is hoped that the nano-system, which combines dual-mode imaging and targeted therapy, will provide a promising strategy for the integration of FL/CT diagnosis and PTT therapy for GnRH-R positive prostate cancer.

Hub Gene Screening Associated with Early Glaucoma: An Integrated Bioinformatics Analysis.

Background: Primary open-angle glaucoma (POAG) is the most common type of glaucoma. The potential influence of some DEGs on the progression of POAG was still incomplete. In this study, we integrated transcriptome data with clinical data to investigate the relationship between them in POAG patients. Methods: The gene expression profile (GSE27276) from Gene Expression Omnibus (GEO) was used to identify DEGs. The LIMMA package of R was used to identify the DEGs (Diboun et al., 2006). The adjusted P values (adj P value) were calculated instead to avoid the appearance of false-positive results. Genes with |log2 fold change (FC)| larger than 1 and adj P value < 0.01 were taken as DEGs between PH and PC samples. GO (Gene Ontology) function and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses of the DEGs were performed. Protein-protein interactions (PPIs) of the DEGs were constructed. Results: A total of 182 DEGs were identified through our analysis, of which 119 genes were upregulated and 63 genes were downregulated. GO enrichment analysis illustrated that these DEGs were mostly enriched into haptoglobin binding, antioxidant activity, and organic acid binding. KEGG enrichment analysis illustrated that these DEGs were mostly enriched into Staphylococcus aureus infection. The most significant module was identified by MCODE consists of 8 DEGs, and BCL11A is the seeded gene. The second most significant module consists of 5 DEGs, and IL1RN is the seeded gene. Conclusion: Our results demonstrate the potential influence of some DEGs on the progression of POAG, providing a comprehensive bioinformatics analysis of the pathogenesis, which may contribute to future investigation into the molecular mechanisms and biomarkers.

A smart hydrogel patch with high transparency, adhesiveness and hemostasis for all-round treatment and glucose monitoring of diabetic foot ulcers.

The treatment and management of diabetic foot ulcers (DFUs) is a pretty intractable problem for clinical nursing. Urgently, the "Black Box" status of the healing process prevents surgeons from providing timely analysis for more effective diagnosis and therapy of the wound. Herein, we designed a transparent monitoring system to treat and manage the DFUs with blood oozing and hard-healing, which resolved the problem of blind management for the other conductive patches. This system was prepared from a conductive hydrogel patch with ultra-high transparence (up to 93.6%), adhesiveness and hemostasis, which is engineered by assembling in situ formed poly(tannic acid) (PTA)-doped polypyrrole (PPy) nanofibrils in the poly(acrylamide-acrylated adenine) (P(AM-Aa)) polymer networks. Significantly, the high transparent conductive hydrogel patch can monitor the wound-healing status visually and effectively promote the healing of DFUs by accelerating hemostasis, improving communication between cells, preventing wound infection, facilitating collagen deposition, and promoting angiogenesis. In addition, the versatile hydrogel patch could realize indirect blood glucose monitoring by detecting the glucose levels on wounds, and further sense the movements with different magnitudes of human body timely. This research may provide a novel strategy in the design of chronic wound dressings for monitoring and treating the wounds synergistically.

Berberine: A Promising Treatment for Neurodegenerative Diseases.

Berberine, as a natural alkaloid compound, is characterized by a diversity of pharmacological effects. In recent years, many researches focused on the role of berberine in central nervous system diseases. Among them, the effect of berberine on neurodegenerative diseases has received widespread attention, for example Alzheimer's disease, Parkinson's disease, Huntington's disease, and so on. Recent evidence suggests that berberine inhibits the production of neuroinflammation, oxidative, and endoplasmic reticulum stress. These effects can further reduce neuron damage and apoptosis. Although the current research has made some progress, its specific mechanism still needs to be further explored. This review provides an overview of berberine in neurodegenerative diseases and its related mechanisms, and also provides new ideas for future research on berberine.

Case report of the first molecular diagnosis of Stickler syndrome with a pathogenic COL2A1 variant in a Mongolia family.

BACKGROUND: Stickler syndrome is a group of connective tissue disorders that can affect eye (myopia, cataract, and retinal detachment), skeleton (spondyloepiphyseal dysplasia and precocious arthritis), craniofacies (midfacial under development and cleft palate), and inner ear (conductive and sensorineural); with the degree of symptoms varying among patients. Mutations in the COL2A1, COL11A1, COL11A2, COL9A1, COL9A2, and COL9A3 procollagen genes cause Stickler syndrome. CASE PRESENTATION: A 16-year-old Mongolian girl approached our clinics with retinal detachment. The proband had vitreous degeneration in both eyes, rhegmatogenous retinal detachment in her right eye, a large area of retina degeneration in her left eye, and coupled with severe myopia. No obvious hearing disorder was found, no abnormalities in bones and joints, and her communication and learning capability were also normal. Further clinical examination showed that the patient's other five family members across three generations had vitreous and retina degenerations. Exome sequencing showed a heterozygous splicing variant in COL2A1 in all patients. CONCLUSIONS: In this case report, a pathogenic splicing variant in the COL2A1 gene was identified in a Mongolian family affected with Stickler syndrome type I by exome sequencing. This heterozygous splicing variant in COL2A1 (NM_001844.4:C.2518-1G>A) that may impair splicing, which was suggested by in silico prediction. Next-generation sequencing is helpful for the differential diagnosis of this clinically variable and genetically heterogeneous disorder.

Inhibitory role of miR-203 in the angiogenesis of mice with pathological retinal neovascularization disease through downregulation of SNAI2.

BACKGROUND: Pathological retinal neovascularization is a disease characterized by abnormal angiogenesis in retina that is a major cause of blindness in humans. Previous reports have highlighted the involvement of microRNAs (miRNAs) in retinal angiogenesis. Therefore, we aimed at exploring the mechanism underlying miR-203 regulating the progression of pathological retinal neovascularization. METHODS: Initially, the mouse model of pathological retinal neovascularization disease was established and the hypoxia-induced human retinal microvascular endothelial cells (HRMECs) were generated. Then, miR-203 and SNAI2 expression in HRMECs and retinal tissues was examined. Subsequently, the effects of miR-203 and SNAI2 on viability, migration, apoptosis and angiogenesis of HRMECs were investigated, with the expression of Bax, Ki-67, MMP-2, MMP-9, VEGF and CD34 measured. Finally, the regulation of miR-203 or SNAI2 on GSK-3ß/ß-catenin pathway was determined through examining the levels of phosphorylated p-GSK-3ß and ß-catenin. RESULTS: Poorly expressed miR-203 and highly expressed SNAI2 were found in HRMECs and retinal tissues of pathological retinal neovascularization. Importantly, overexpressed miR-203 or silencing SNAI2 inhibited viability, migration and angiogenesis but promoted apoptosis of HRMECs, evidenced by elevated Bax expression but reduced expression of Ki-67, MMP-2, MMP-9, VEGF and CD34. Moreover, overexpression of miR-203 was found to repress the GSK-3ß/ß-catenin pathway by downregulating SNAI2. CONCLUSION: Collectively, this study demonstrated that overexpression of miR-203 suppressed the angiogenesis in mice with pathological retinal neovascularization disease via the inactivation of GSK-3ß/ß-catenin pathway by inhibiting SNAI2, which provided a novel therapeutic insight for pathological retinal neovascularization disease.

Robust Fovea Localization Based on Symmetry Measure.

Automatic fovea localization is a challenging issue. In this article, we focus on the study of fovea localization and propose a robust fovea localization method. We propose concentric circular sectional symmetry measure (CCSSM) for symmetry axis detection, and region of interest (ROI) determination, which is a global feature descriptor robust against local feature changes, to solve the lesion interference issue, i.e., fovea visibility interference from lesions, using both structure features and morphological features. We propose the index of convexity and concavity (ICC) as the convexity-concavity measure of the surface and provide a quantitative evaluation tool for ophthalmologists to learn whether the occurrence of lesion within the ROI. We propose the weighted gradient accumulation map, which is insensitive to local intensity changes and can overcome the influence of noise and contamination, to perform refined localization. The advantages of the proposed method lies in two aspects. First, the accuracy and robustness can be achieved without typical sophisticated manner, i.e., blood vessel segmentation and parabola fitting. Second, the lesion interference is considered in our plan of fovea localization. Our proposed symmetry-based method is innovative in the solution of fovea detection, and it is simple, practical, and controllable. Experiment results show that the proposed method can resist the interference of unbalanced illumination and lesions, and achieve high accuracy rate in five datasets. Compared to the state-of-the-art methods, high robustness and accuracy of the proposed method guarantees its reliability.

Parthenolide inhibits the proliferation and induces the apoptosis of human uveal melanoma cells.

AIM: To explore the effect of parthenolide (PTL) on human uveal melanoma (UM) cells (C918 and SP6.5 cells) and its molecular mechanism. METHODS: Carboxyfluorescein succinimidyl amino ester (CFSE) assays and cell counting kit-8 (CCK-8) were performed to detect the cell viability. Flow cytometry was used to analyze cell cycle and apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assays were performed to measure proliferation-related and apoptosis-related factors. RESULTS: Firstly, PTL decreased the viability of C918 and SP6.5 cells in a dose-dependent manner, and the effect of PTL on C918 cells was stronger than on SP6.5; however, it did not affect normal cells. Secondly, PTL increased the proportion of cell number at cell cycle G1 phase in C918 cells, and decreased the proportion of cell number at S phase, but the proportion did not change at G2 phase. In addition, PTL induced the apoptosis of C918 cells, and decreased the expressions of Cyclin D1, B-cell lymphoma-2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-XL). Also, PTL increased Cyclin inhibition protein 1 (P21), Bcl-2-associated X protein (Bax), Cysteinyl aspartate specific proteinas-3 (Caspase-3) and Caspase-9 expression. However, the expression of Caspase-8 was not changed. CONCLUSION: PTL inhibites proliferation and induces apoptosis in UM cells by arresting G1 phase and regulating mitochondrial pathway, however, it does not affect normal cells.

One-step fabrication of boronic-acid-functionalized carbon dots for the detection of sialic acid.

Typically, sialic acids (SA) with a nine-carbon backbone are found at the glycan chain termini on the cell membranes, which play crucial roles in various physiological and pathological processes. The expression level of SA in the blood serum has been reported to correlate with various disease states among cancer. In this study, a novel approach for preparing fluorescent boronic-acid-modified carbon dots (C-dots) for the detection of SA was developed. The functionalized C-dots were synthesized by a facile, one-step hydrothermal method using 3-pyridineboronic acid as the sole carbon source. The added SA selectively recognized the C-dots, leading to the fluorescence quenching of the C-dots in a linear range of 80-4000 µM with a detection limit of 54 µM. The as-developed boronic-acid nanoprobe was successfully applied for the detection of SA in human serum samples with satisfactory results. In addition, this method afforded results within 4 min. Compared to other methods, this new proposed approach was simpler and exhibited excellent sensitivity and selectivity, demonstrating immense potential as an alternative for SA detection.

Knockdown of long noncoding RNA 00152 (LINC00152) inhibits human retinoblastoma progression.

BACKGROUND: A growing body of evidence supports the involvement of long noncoding RNA 00152 (LINC00152) in the progression and metastasis of multiple cancers. However, the exact roles of LINC00152 in the progression of human retinoblastoma (RB) remain unknown. We explored the expression and biological function of human RB. MATERIALS AND METHODS: The expression level of LINC00152 in RB tissues and cells was analyzed using quantitative real-time PCR. The function of LINC00152 was determined using a series of in vitro assays. In vivo, a nude mouse model was established to analyze the function of LINC00152. Gene and protein expressions were detected using quantitative real-time PCR and Western blot assays, respectively. RESULTS: The expression of LINC00152 mRNA was upregulated in RB tissues and cell lines. Knockdown of LINC00152 significantly inhibited cell proliferation, colony formation, migration, and invasion and promoted cell apoptosis and caspase-3 and caspase-8 activities in vitro, as well as suppressing tumorigenesis in vivo. We identified several genes related to proliferation, apoptosis, and invasion including Ki-67, Bcl-2, and MMP-9 that were transcriptionally inactivated by LINC00152. CONCLUSION: Taken together, these data implicate LINC00152 as a therapeutic target in RB.

FBLN4 as candidate gene associated with long-term and short-term survival with primary glioblastoma.

BACKGROUND: Glioblastoma multiforme (GBM) is the most common malignant and lethal type of primary central nervous system tumor in humans. In spite of its high lethality, a small percentage of patients have a relatively good prognosis, with median survival times of 36 months or longer. The identification of clinical subsets of GBM associated with distinct molecular genetic profiles has made it possible to design therapies tailored to treat individual patients. METHODS: We compared microarray data sets from long-term survivors (LTSs) and short-term survivors (STSs) to screen for prognostic biomarkers in GBM patients using the WebArrayDB platform. We focused on FBLN4, IGFBP-2, and CHI3L1, all members of a group of 10 of the most promising, differentially regulated gene candidates. Using formalin-fixed paraffin-embedded GBM samples, we corroborated the relationship between these genes and patient outcomes using methylation-specific polymerase chain reaction (PCR) for MGMT methylation status and quantitative reverse transcription PCR for expression of these genes. RESULTS: Expression levels of the mRNAs of these 3 genes were higher in the GBM samples than in normal brain samples and these 3 genes were significantly upregulated in STSs compared to the levels in LTS samples (P<0.01). Furthermore, Kaplan-Meier analysis showed that the expression patterns of FBLN4 and IGFBP-2 serve as independent prognostic indicators for overall survival (P<0.01 and P<0.05, respectively). CONCLUSION: To our knowledge, this is the first report describing FBLN4 as a prognostic factor for GBM patient survival, demonstrating that increased GBM survival time correlates with decreased FBLN4 expression. Understanding FBLN4 expression patterns could aid in the creation of powerful tools to predict clinical prognoses of GBM patients.