Dual mRNA co-delivery for in situ generation of phagocytosis-enhanced CAR macrophages augments hepatocellular carcinoma immunotherapy.

Hepatocellular carcinoma (HCC) is a prevalent and lethal disease, and tumor regression rarely occurs in advanced HCC patients due to limited effective therapies. Given the enrichment of macrophages in HCC and their role in tumor immunity, transforming them into chimeric antigen receptor macrophages (CAR-Ms) is thought to increase HCC cell-directed phagocytosis and tumoricidal immunity. To test this hypothesis, mRNA encoding CAR is encapsulated in a lipid nanoparticle (LNP) that targets liver macrophages. Notably, the LNPs adsorb specific plasma proteins that enable them to target HCC-associated macrophages. Moreover, mRNA encoding Siglec-G lacking ITIMs (Siglec-GΔITIMs) is codelivered to liver macrophages by LNP to relieve CD24-mediated CAR-Ms immune suppression. Mice treated with LNPs generating CAR-Ms as well as CD24-Siglec-G blockade significantly elevate the phagocytic function of liver macrophages, reduce tumor burden and increase survival time in an HCC mouse model. Arguably, our work suggests an efficacious and flexible strategy for the treatment of HCC and warrants further rigorous evaluation in clinical trials.

Health-related quality of life in patients with Kashin-Beck disease is lower than in those with osteoarthritis: a cross-sectional study.

BACKGROUND: Kashin-Beck disease (KBD) is an endemic deformable bone and joint disease, which affects the quality of life (QOL) of patients. We conducted a cross-sectional study of the QOL of KBD patients by a new KBD quality of life (KBDQOL) questionnaire. METHODS: A total of 252 KBD patients and 248 OA patients came from Northwest China, and 260 healthy people living in the same area as KBD and osteoarthritis (OA) patients served as the controls. KBDQOL questionnaire was used to evaluate the QOL of all objects. RESULTS: The average scores for physical functions, activity limitations, support of society, mental health and general health were significantly lower in KBD patients than that in OA patients and healthy people except for economics. Monofactor analysis showed that age, height, weight status, education level and grade of KBD had a significant effect on KBDQOL score. Multivariate analysis showed that grade of KBD was the influencing factor of physical function score; gender, age, height, grade of KBD and duration of symptoms were the influencing factors of activity restriction score; age and grade of KBD were factors affecting the general health score. CONCLUSION: The QOL of KBD patients was significantly lower than that of OA patients and healthy people. The KBDQOL questionnaire may be a promising tool for assessing the QOL of KBD patients.

Application of a Novel Au@ZIF-8 Composite in the Detection of Bisphenol A by Surface-Enhanced Raman Spectroscopy.

Bisphenol A (BPA) is an endocrine disruptor which is widely present in fish under the influence of environmental pollution. It is essential to establish a rapid detection method for BPA. Zeolitic imidazolate framework (ZIF-8) is a typical metal-organic framework material (MOFs) with a strong adsorption capacity, which can effectively adsorb harmful substances in food. Combining MOFs and surface-enhanced Raman spectroscopy (SERS) can achieve rapid and accurate screening of toxic substances. In this study, a rapid detection method for BPA was established by preparing a new reinforced substrate Au@ZIF-8. The SERS detection method was optimized by combining SERS technology with ZIF-8. The Raman peak at 1172 cm-1 was used as the characteristic quantitative peak, and the lowest detection concentration of BPA was as low as 0.1 mg/L. In the concentration range of 0.1~10 mg/L, the linear relationship between SERS peak intensity and the concentration of BPA was good, and R2 was 0.9954. This novel SERS substrate was proven to have great potential in rapidly detecting BPA in food.

Impact of Whole-Layer Barbed Suture Closure on the Postoperative Effect and Aesthetic Satisfaction with Incision After Knee Arthroplasty.

Objective: To evaluate the effect of whole-layer barbed suture for incision closure on the clinical outcome and aesthetic satisfaction of patients with the incision following total knee arthroplasty (TKA). Materials and Methods: A retrospective analysis was conducted on 94 patients (37 men and 57 women, 50-84 years old) who underwent a first TKA between May 2018 and April 2021. The enrolled patients were divided into two groups according to the suture mode, with 45 cases (group A) receiving closure of the deep tissue with a barbed suture and intradermal suture of the incision with another barbed suture and 49 cases (group B) receiving closure of the deep tissue with absorbable suture and interrupted suture of the incision with nonabsorbable suture. Further comparisons were performed regarding incision length, suture time, postoperative incision complications, Hollander Wound Evaluation Score (HWES), Hospital for Special Surgery (HSS) knee score, American Knee Society (AKS) score, and Patient and Observer Scar Assessment Scale (POSAS). The clinical effects of the two suture modes in TKA and the patients' aesthetic satisfaction with the incision were also evaluated at 2 weeks, 3 months and 6 months after the operation. Results: Compared with group B, group A had higher HWES at 2 weeks after the operation (P < 0.05), shorter suture times (P < 0.05) and lower POSAS scores at 3 and 6 months after the operation (P < 0.05). However, no significant difference was observed between the two groups in incision length, complication rate, HSS or AKS scores (P > 0.05). Conclusion: Compared with the interrupted suture, the use of whole-layer barbed suture for incision closure after TKA has the advantages of a safe and effective outcome, short operation time, high cosmesis degree of the wound scar and high patient satisfaction.

Cross-sectional analysis of risk factors for surgical site infection secondary to spinal internal fixation via the posterior approach.

OBJECTIVE: This study was performed to investigate the risk factors related to surgical site infection (SSI) secondary to spinal internal fixation via the posterior approach. METHODS: Patients who had undergone spinal internal fixation via the posterior approach were selected for inclusion in this cross-sectional study. Factors related to SSI were analysed using univariate and multivariate analyses. RESULTS: Among 4,350 patients, 66 had SSI (infection prevalence of 1.5%). Multivariate logistic regression analysis demonstrated that age of >60 years, surgical duration of >3 hours, haemoglobin concentration of <80 g/L, serum albumin concentration of <30 g/L, diabetes history, and blood loss of >1,000 mL were significantly correlated with SSI secondary to spinal internal fixation via the posterior approach. CONCLUSIONS: This study provides information on SSI secondary to spinal internal fixation via the posterior approach. We found that age of >60 years, surgical duration of >3 hours, haemoglobin concentration of <80 g/L, serum albumin concentration of <30 g/L, diabetes history, and blood loss of >1,000 mL are directly correlated with SSI secondary to spinal internal fixation via the posterior approach.

Three-dimensional morphometric differences of resected distal femurs and proximal tibias in osteoarthritic and normal knees.

BACKGROUND: There is a paucity of data concerning the morphological differences of resected distal femurs and proximal tibias in osteoarthritic (OA) and normal knees. The objective of this study was to determine whether morphometric differences in the surfaces of resected distal femurs and proximal tibias exist between OA and normal knees in a Chinese population. METHODS: Ninety-four OA knees and ninety-five normal knees were evaluated in Chinese individuals. Computed tomography was used to measure the femoral mediolateral (fML), medial anteroposterior (fMAP), lateral anteroposterior (fLAP), medial condylar width (fMCW), lateral condylar width (fLCW), medial posterior condylar curvature radii (fMCR), lateral posterior condyle curvature radii (fLCR), fML/fMAP aspect ratio, tibial mediolateral (tML), middle anteroposterior (tAP), medial anteroposterior (tMAP), and lateral anteroposterior (tLAP) tML/tMAP aspect ratio to determine the morphologic differences between OA and normal knees. RESULTS: The average fMCW and tMAP dimensions of OA knees were larger than those of normal knees in both male and female (p <0.05). The fMAP/fML aspect ratio and tMAP/tML aspect ratio were also significantly different in both sexs (p <0.05). OA knees have an oval-shaped distal femur with a wider ML length and more spherical-shaped proximal tibiae with relatively narrow ML dimensions. CONCLUSIONS: The study revealed the morphological differences in fMCW, tMAP, fMAP/fML and tMAP/tML between OA and normal knees in both males and females. These findings may provide guidelines that can be used to design better knee implants that are more size-matched for OA knees.

Long-term outcomes of arthroscopic debridement of the knee in adults with Kashin-Beck disease: an 18-year follow-up.

OBJECTIVE: Kashin-Beck disease (KBD) is an endemic degenerative joint disease with a high disability rate. We retrospectively evaluated the 18-year clinical follow-up outcomes of adult patients with KBD who underwent arthroscopic debridement for knee osteoarthritis. METHODS: Thirty-one patients with KBD (31 knees) underwent arthroscopy for knee osteoarthritis. The visual analog scale (VAS) score, walking distance, knee mobility, and patients' self-evaluated improvement in clinical symptoms were retrospectively evaluated before and 18 years after the operation. RESULTS: The patients' self-evaluated clinical symptoms showed considerable improvement at 2, 6, and 8 years after surgery but deteriorated at 10 and 18 years after surgery. Knee mobility was greater after than before arthroscopy but decreased from 6 to 18 years postoperatively. The VAS score for knee pain was high before the operation, decreased at 2 years postoperatively, increased at 6 years postoperatively, and was significantly lower at 18 years postoperatively than before surgery. The walking distance was significantly longer at 2, 6, and 8 years postoperatively than preoperatively. CONCLUSIONS: Arthroscopic treatment may be an effective therapy for adult patients with KBD who develop knee osteoarthritis. In this study, arthroscopy had a long-term effect on patients with KBD who had Kellgren-Lawrence grade

Pan-Cancer Analysis of Prognostic and Immune Infiltrates for CXCs.

BACKGROUND: CXCs are important genes that regulate inflammation and tumor metastasis. However, the expression level, prognosis value, and immune infiltration of CXCs in cancers are not clear. METHODS: Multiple online datasets were used to analyze the expression, prognosis, and immune regulation of CXCs in this study. Network analysis of the Amadis database and GEO dataset was used to analyze the regulation of intestinal flora on the expression of CXCs. A mouse model was used to verify the fact that intestinal bacterial dysregulation can affect the expression of CXCs. RESULTS: In the three cancers, multiple datasets verified the fact that the mRNA expression of this family was significantly different; the mRNA levels of CXCL3, 8, 9, 10, 14, and 17 were significantly correlated with the prognosis of three cancers. CXCs were correlated with six types of immuno-infiltrating cells in three cancers. Immunohistochemistry of clinical samples confirmed that the expression of CXCL8 and 10 was higher in three cancer tissues. Animal experiments have shown that intestinal flora dysregulation can affect CXCL8 and 10 expressions. CONCLUSION: Our results further elucidate the function of CXCs in cancers and provide new insights into the prognosis and immune infiltration of breast, colon, and pancreatic cancers, and they suggest that intestinal flora may influence disease progression through CXCs.

Ten-Eleven Translocation 1 Promotes Malignant Progression of Cholangiocarcinoma With Wild-Type Isocitrate Dehydrogenase 1.

BACKGROUND AND AIMS: Cholangiocarcinoma (CCA) is a highly lethal disease without effective therapeutic approaches. The whole-genome sequencing data indicate that about 20% of patients with CCA have isocitrate dehydrogenase 1 (IDH1) mutations, which have been suggested to target 2-oxoglutarate (OG)-dependent dioxygenases in promoting CCA carcinogenesis. However, the clinical study indicates that patients with CCA and mutant IDH1 have better prognosis than those with wild-type IDH1, further complicating the roles of 2-OG-dependent enzymes. APPROACH AND RESULTS: This study aimed to clarify if ten-eleven translocation 1 (TET1), which is one of the 2-OG-dependent enzymes functioning in regulating 5-hydroxymethylcytosine (5hmC) formation, is involved in CCA progression. By analyzing The Cancer Genome Atlas (TCGA) data set, TET1 mRNA was found to be substantially up-regulated in patients with CCA when compared with noncancerous bile ducts. Additionally, TET1 protein expression was significantly elevated in human CCA tumors. CCA cells were challenged with α-ketoglutarate (α-KG) and dimethyl-α-KG (DM-α-KG), which are cosubstrates for TET1 dioxygenase. The treatments with α-KG and DM-α-KG promoted 5hmC formation and malignancy of CCA cells. Molecular and pharmacological approaches were used to inhibit TET1 activity, and these treatments substantially suppressed 5hmC and CCA carcinogenesis. Mechanistically, it was found that knockdown of TET1 may suppress CCA progression by targeting cell growth and apoptosis through epigenetic regulation. Consistently, targeting TET1 significantly inhibited CCA malignant progression in a liver orthotopic xenograft model by targeting cell growth and apoptosis. CONCLUSIONS: Our data suggest that expression of TET1 is highly associated with CCA carcinogenesis. It will be important to evaluate TET1 expression in CCA tumors before application of the IDH1 mutation inhibitor because the inhibitor suppresses 2-hydroxyglutarate expression, which may result in activation of TET, potentially leading to CCA malignancy.

Automatic deep learning-based colorectal adenoma detection system and its similarities with pathologists.

OBJECTIVES: The microscopic evaluation of slides has been gradually moving towards all digital in recent years, leading to the possibility for computer-aided diagnosis. It is worthwhile to know the similarities between deep learning models and pathologists before we put them into practical scenarios. The simple criteria of colorectal adenoma diagnosis make it to be a perfect testbed for this study. DESIGN: The deep learning model was trained by 177 accurately labelled training slides (156 with adenoma). The detailed labelling was performed on a self-developed annotation system based on iPad. We built the model based on DeepLab v2 with ResNet-34. The model performance was tested on 194 test slides and compared with five pathologists. Furthermore, the generalisation ability of the learning model was tested by extra 168 slides (111 with adenoma) collected from two other hospitals. RESULTS: The deep learning model achieved an area under the curve of 0.92 and obtained a slide-level accuracy of over 90% on slides from two other hospitals. The performance was on par with the performance of experienced pathologists, exceeding the average pathologist. By investigating the feature maps and cases misdiagnosed by the model, we found the concordance of thinking process in diagnosis between the deep learning model and pathologists. CONCLUSIONS: The deep learning model for colorectal adenoma diagnosis is quite similar to pathologists. It is on-par with pathologists' performance, makes similar mistakes and learns rational reasoning logics. Meanwhile, it obtains high accuracy on slides collected from different hospitals with significant staining configuration variations.

[Caspase-9 (-Ex5+32 G/A) gene polymorphism is associated with caspase-9 expression in degenerative nucleus pulposus of lumbar intervertebral disc].

Objective To investigate the relationship between caspase-9 (-Ex5+32 G/A) gene polymorphisms, Schneiderman score of magnetic resonance imaging (MRI) in lumbar disc degeneration and the expression of caspase-9 in degenerative nucleus pulposus. Methods The peripheral venous blood and prominent nucleus pulposus were obtained from 105 patients with lumbar disc herniation. Genomic DNA was extracted and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The caspase-9(-Ex5+32 G/A) gene polymorphisms were analyzed in all DNA samples. The expression of caspase-9 in the tissues was detected by immunohistochemical SP staining. The t test was used to analyze the difference between the genotypes and the expression of caspase-9 in intervertebral disc nucleus. Pearson correlation analysis was performed to assess the association between caspase-9 expression in nucleus pulposus and the MRI score of lumbar disc. Results MRI analysis showed that the patients with AA genotype had the highest MRI scores, but there were no significant differences in MRI scores among patients with AA, GA, and GG genotypes. Compared with GG genotype carriers, AA genotype carriers had a statistically significant difference in the expression of caspase-9 in nucleus pulposus. There was no correlation between the MRI score of lumbar disc degeneration and the expression of caspase-9 in the degenerated nucleus pulposus. Conclusion The caspase-9(-Ex5+32 G/A) gene polymorphism is associated with the expression of caspase-9 in the degenerative nucleus pulposus. However, the MRI Schneiderman score is not significantly correlated with the expression of caspase-9 in degenerative nucleus pulposus.

A radial calibration window for analytical ultracentrifugation.

Analytical ultracentrifugation (AUC) is a first-principles based method for studying macromolecules and particles in solution by monitoring the evolution of their radial concentration distribution as a function of time in the presence of a high centrifugal field. In sedimentation velocity experiments, hydrodynamic properties relating to size, shape, density, and solvation of particles can be measured, at a high hydrodynamic resolution, on polydisperse samples. In a recent multilaboratory benchmark study including data from commercial analytical ultracentrifuges in 67 laboratories, the calibration accuracy of the radial dimension was found to be one of the dominant factors limiting the accuracy of AUC. In the present work, we develop an artifact consisting of an accurately calibrated reflective pattern lithographically deposited onto an AUC window. It serves as a reticle when scanned in AUC control experiments for absolute calibration of radial magnification. After analysis of the pitch between landmarks in scans using different optical systems, we estimate that the residual uncertainty in radial magnification after external calibration with the radial scale artifact is ≈0.2 %, of similar magnitude to other important contributions after external calibration such as the uncertainty in temperature and time. The previous multilaboratory study had found many instruments with errors in radial measurements of 1 % to 2 %, and a few instruments with errors in excess of 15 %, meaning that the use of the artifact developed here could reduce errors by 5-to 10-fold or more. Adoption of external radial calibration is thus an important factor for assuring accuracy in studies related to molecular hydrodynamics and particle size measurements by AUC.

[Association of FasL-844T/C gene polymorphism with FasL expression in the nucleus pulposus of degenerative lumbar intervertebral discs].

OBJECTIVE: To investigate the association of FasL-844T/C gene polymorphism with the magnetic resonance imaging (MRI) findings and FasL expression in the nucleus pulposus of degenerative lumbar intervertebral discs. METHODS: Lumbar MRI data, venous blood and nucleus pulposus were collected from 105 patients with lumbar disc herniation. The genotypes of FasL-844T/C gene of the patients were determined using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Immunohistochemistry was used to detect the expression of FasL in the nucleus pulposus of the degenerative lumbar intervertebral discs. RESULTS: Compared with CC genotype, TT genotype of FasL-844T/C gene was associated with a significantly increased score of lumbar disc degeneration (P=0.003) as observed in MRI scan. FasL expression in the nucleus pulposus differed significantly between patients of FASL-844CC genotype and those of FASL-844TT genotype (P=0.048), but not between those of FASL-844CC and FASL-844CT genotypes (P=0.264). No significant association was found between MRI findings and FasL expression in the nucleus pulposus of the lumbar intervertebral discs. CONCLUSION: FasL-844T/C gene polymorphism is correlated with the expression of FasL in the nucleus pulposus of the intervertebral disc in patients with lumbar disc herniation. MRI findings of the lumbar intervertebral discs do not correlate with the expression of FasL in the nucleus pulposus of the intervertebral discs.

Acceleration of tendon-bone healing of anterior cruciate ligament graft using intermittent negative pressure in rabbits.

BACKGROUND: The purpose of this study was to test effects of negative pressure on tendon-bone healing after reconstruction of anterior cruciate ligament (ACL) in rabbits. METHODS: Hind legs of 24 New Zealand White rabbits were randomly selected as negative pressure group and the contralateral hind legs as control. Reconstruction of the ACL was done. Joints of the negative pressure side were placed with drainage tubes connecting the micro-negative pressure aspirator. Control side was placed with ordinary drainage tubes. Drainage tubes on both sides were removed at the same time 5 days after operation. After 6 weeks, joint fluid was drawn to detect the expression levels of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α); at the same time, femur-ligament-tibia complex was obtained to determine tendon graft tension and to observe the histomorphology, blood vessels of the tendon-bone interface, and expression of vascular endothelial growth factor (VEGF). RESULTS: The maximum load breakage of tendon graft was significantly greater in the negative pressure group than in the control group (P < 0.05). Histological studies of the tendon-bone interface found that there was more new bone formation containing chondroid cells and aligned connective tissue in the negative pressure group than in the control group. Expression of VEGF was higher in the negative pressure group than in the control group (P < 0.01). Content of IL-1ß and TNF-α in synovial fluid is lower in the negative pressure group than in the control group (P < 0.01). CONCLUSIONS: Intermittent negative pressure plays an active role in tendon-bone healing and creeping substitution of ACL reconstruction in the rabbits.

Staged total knee arthroplasty for bilateral complex knee deformities from Kashin-Beck disease and skeletal dysplasia.

This study reported two cases of patients with Grade III Kashin-Beck disease (KBD) with skeletal dysplasia concomitant with complex knee deformity and functional limitation treated by staged total knee arthroplasty (TKA). Detailed pre-operative planning, bone resection, and soft tissue balancing in affected knees were performed in the surgeries in this report. The results demonstrated that TKA could correct lower limb alignment, alleviate knee pain, improve function, and provide good quality of life in people with KBD. Surgical efficacy is still lower compared with treatment for osteoarthritis; contributing factors include weak muscle strength, severe deformity and unequal length of the lower limb, weak extensor apparatus of the knee, and patient-specific factors.

Eosinophilic pancreatitis: Three case reports and literature review.

Eosinophilic pancreatitis (EP) is a rare form of chronic pancreatitis characterized by localized or diffuse eosinophilic infiltration of the pancreas and elevated serum immunoglobulin E levels. EP is difficult to distinguish from pancreatic cancer on the basis of clinical symptoms and the results of auxiliary examination alone. A retrospective analysis of the clinicopathological characteristics and laboratory, imaging, and pathology results of 3 patients with EP, who were initially diagnosed with pancreatic malignancy, was performed. EP is an allergic disease with non-specific clinical manifestations that is difficult to distinguish from pancreatic cancer based exclusively on clinical symptoms and auxiliary examination, resulting in the need for invasive procedures to confirm the diagnosis. An increase in the eosinophil count in the peripheral blood and pathological examination are essential for the diagnosis of EP.

Long-Term Efficacy of Arthroscopic Debridement on Knee Osteoarthritis in Patients with Kashin-Beck Disease.

Kashin-Beck Disease (KBD) is a chronic and degenerative joint disease with decreased health conditions causing major functional limitations and disability. In this study, we investigated the long-term clinical outcomes of arthroscopic debridement for knee osteoarthritis in KBD patients with Kellgren-Lawrence I-III classification. 31 KBD patients underwent arthroscopic debridement and 24 of them were followed up 8 years after the surgery. The Lysholm scores, femorotibial angle (FTA), and joint space angle (JSA) of the knees were assessed and compared preoperatively and postoperatively to evaluate the clinic outcome. Our results showed that the Lysholm scores, but not the FTA or JSA, of these patients were significantly improved 8 years after the therapy, suggesting that arthroscopic surgery has long-term efficacy in KBD patients with Kellgren-Lawrence I-III classification.

PGE2 receptor agonist misoprostol protects brain against intracerebral hemorrhage in mice.

Intracerebral hemorrhage (ICH) is a devastating form of stroke. Misoprostol, a synthetic prostaglandin E1 (PGE1) analog and PGE2 receptor agonist, has shown protection against cerebral ischemia. In this study, we tested the efficacy of misoprostol in the 12-month-old mice subjected to 1 of 2 complementary ICH models, the collagenase model (primary study) and blood model (secondary study, performed in an independent laboratory). We also investigated its potential mechanism of action. Misoprostol posttreatment decreased brain lesion volume, edema, and brain atrophy and improved long-term functional outcomes. In the collagenase-induced ICH model, misoprostol decreased cellular inflammatory response; attenuated oxidative brain damage and gelatinolytic activity; and decreased high-mobility group box 1 (HMGB1) expression, Src kinase activity, and interleukin-1ß expression without affecting cyclooxygenase-2 expression. Furthermore, HMGB1 inhibition with glycyrrhizin decreased Src kinase activity, gelatinolytic activity, neuronal death, and brain lesion volume. Src kinase inhibition with 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) decreased gelatinolytic activity and brain edema and improved neurologic function but did not decrease HMGB1 protein level. These results indicate that misoprostol protects brain against ICH injury through mechanisms that may involve the HMGB1, Src kinase, and matrix metalloproteinase-2/9 pathways.

Reduction of α-dystroglycan expression is correlated with poor prognosis in glioma.

Dystroglycan (DG), a multifunctional protein dimer of non-covalently linked α and ß subunits, is best known as an adhesion and transduction molecule linking the cytoskeleton and intracellular signaling pathways to extracellular matrix proteins. Loss of DG binding, possibly by degradation or disturbed glycosylation, has been reported in a variety of cancers. DG is abundant at astroglial endfeet forming the blood-brain barrier (BBB) and glia limitans; so, we examined if loss of expression is associated with glioma. Expression levels of α-DG and ß-DG were assessed by immunohistochemistry in a series of 78 glioma specimens to determine the relationship with tumor grade and possible prognostic significance. α-DG immunostaining was undetectable in 44 of 49 high-grade specimens (89.8%) compared to 15 of 29 low-grade specimens (51.72%) (P<0.05). Moreover, loss of α-DG expression was an independent predictor of shorter disease-free survival (DFS) (hazards ratio (HR) = 0.142, 95% confidence interval (CI) 0.033-0.611, P=0.0088). Reduced expression of both α-DG and ß-DG was also a powerful negative prognostic factor for DFS (HR=2.556, 95% CI 1.403-4.654, P=0.0022) and overall survival (OS) (HR=2.193, 95% CI 1.031-4.666, P=0.0414). Lack of α-DG immunoreactivity is more frequent in high-grade glioma and is an independent predictor of poor clinical outcome. Similarly, lack of both α-DG and ß-DG immunoreactivity is a strong independent predictor of clinical outcome.