Unraveling T cell exhaustion in the immune microenvironment of osteosarcoma via single-cell RNA transcriptome.

Osteosarcoma (OS) represents a profoundly invasive malignancy of the skeletal system. T cell exhaustion (Tex) is known to facilitate immunosuppression and tumor progression, but its role in OS remains unclear. In this study, single-cell RNA sequencing data was employed to identify exhausted T cells within the tumor immune microenvironment (TIME) of OS. We found that exhausted T cells exhibited substantial infiltration in OS samples. Pseudotime trajectory analysis revealed a progressive increase in the expression of various Tex marker genes, including PDCD1, CTLA4, LAG3, ENTPD1, and HAVCR2 in OS. GSVA showed that apoptosis, fatty acid metabolism, xenobiotic metabolism, and the interferon pathway were significantly activated in exhausted T cells in OS. Subsequently, a prognostic model was constructed using two Tex-specific genes, MYC and FCGR2B, which exhibited exceptional prognostic accuracy in two independent cohorts. Drug sensitivity analysis revealed that OS patients with a low Tex risk were responsive to Dasatinib and Pazopanib. Finally, immunohistochemistry verified that MYC and FCGR2B were significantly upregulated in OS tissues compared with adjacent tissues. This study investigates the role of Tex within the TIME of OS, and offers novel insights into the mechanisms underlying disease progression as well as the potential treatment strategies for OS.

Calcium phosphate coating enhances osteointegration of melt electrowritten scaffold by regulating macrophage polarization.

The osteoimmune microenvironment induced by implants plays a significant role in bone regeneration. It is essential to efficiently and timely switch the macrophage phenotype from M1 to M2 for optimal bone healing. This study examined the impact of a calcium phosphate (CaP) coating on the physiochemical properties of highly ordered polycaprolactone (PCL) scaffolds fabricated using melt electrowritten (MEW). Additionally, it investigated the influence of these scaffolds on macrophage polarization and their immunomodulation on osteogenesis. The results revealed that the CaP coated PCL scaffold exhibited a rougher surface topography and higher hydrophilicity in comparison to the PCL scaffold without coating. Besides, the surface morphology of the coating and the release of Ca2+ from the CaP coating were crucial in regulating the transition of macrophages from M1 to M2 phenotypes. They might activate the PI3K/AKT and cAMP-PKA pathways, respectively, to facilitate M2 polarization. In addition, the osteoimmune microenvironment induced by CaP coated PCL could not only enhance the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in vitro but also promote the bone regeneration in vivo. Taken together, the CaP coating can be employed to control the phenotypic switching of macrophages, thereby creating a beneficial immunomodulatory microenvironment that promotes bone regeneration.

Gut microbiota modification by diosgenin mediates antiepileptic effects in a mouse model of epilepsy.

Diosgenin, a natural steroid saponin, holds promise as a multitarget therapeutic for various diseases, including neurodegenerative conditions. Its efficacy in slowing Alzheimer's disease, Parkinson's disease, multiple sclerosis, and stroke progression has been demonstrated. However, the role of diosgenin in anti-epilepsy and its potential connection to the modulation of the intestinal microbiota remain poorly understood. In this study, exogenous diosgenin significantly mitigated pentylenetetrazole (PTZ)-induced seizures, learning and memory deficits, and hippocampal neuronal injury. 16S ribosomal RNA (16S rRNA) sequencing revealed a reversal in the decrease of Bacteroides and Parabacteroides genera in the PTZ-induced mouse epileptic model following diosgenin treatment. Fecal microbiota transplantation (FMT) experiments illustrated the involvement of diosgenin in modulating gut microbiota and providing neuroprotection against epilepsy. Our results further indicated the repression of enteric glial cells (EGCs) activation and the TLR4-MyD88 pathway, coupled with reduced production of inflammatory cytokines in the colonic lumen, and improved intestinal barrier function in epilepsy mice treated with diosgenin or FMT. This study suggests that diosgenin plays a role in modifying gut microbiota, contributing to the alleviation of intestinal inflammation and neuroinflammation, ultimately inhibiting epilepsy progression in a PTZ-induced mouse model. Diosgenin emerges as a potential therapeutic option for managing epilepsy and its associated comorbidities.

Characterization of myeloid signature genes for predicting prognosis and immune landscape in Ewing sarcoma.

Myeloid cells as a highly heterogeneous subpopulation of the tumor microenvironment (TME) are intimately associated with tumor development. Ewing sarcoma (EWS) is characterized by abundant myeloid cell infiltration in the TME. However, the correlation between myeloid signature genes (MSGs) and the prognosis of EWS patients was unclear. In this research, we synthetically characterized the expression of MSGs in a training cohort and classified EWS patients into two subtypes. Immune cell infiltration analysis revealed that MSGs subtypes correlated closely with different immune statuses. Furthermore, a three-gene prognostic model (CTSD, SIRPA, and FN1) was constructed by univariate, LASSO, and multivariate Cox analysis, and it showed excellent prognostic accuracy in EWS patients. We also developed a nomogram for better predicting the long-term survival of EWS. Functional enrichment analysis showed immune-related pathways were distinctly different in the high- and low-risk groups. Further analysis revealed that patients in the high-risk group were tightly associated with an immunosuppressive microenvironment. Finally, we validated the expression of these candidate genes by Western blot (WB), qPCR, and immunohistochemistry (IHC) analysis. To sum up, our study identified that the MSGs model was strongly linked to prognostic prediction and immune infiltration in EWS patients, providing novel insights into the clinical treatment and management of EWS patients.

Blood transcriptome and machine learning identified the crosstalk between COVID-19 and fibromyalgia: a preliminary study.

OBJECTIVES: The COVID-19 pandemic caused by SARS-CoV-2 has seriously threatened the human health. Growing evidence shows that COVID-19 patients who recovery will persist with symptoms of fibromyalgia (FM). However, the common molecular mechanism between COVID-19 and FM remains unclear. METHODS: We obtained blood transcriptome data of COVID-19 (GSE177477) and FM (GSE67311) patients from GEO database, respectively. Subsequently, we applied Limma, GSEA, Wikipathway, KEGG, GO, and machine learning analysis to confirm the common pathogenesis between COVID-19 and FM, and screened key genes for the diagnosis of COVID-19 related FM. RESULTS: A total of 2505 differentially expressed genes (DEGs) were identified in the FM dataset. Functional enrichment analysis revealed that the occurrence of FM was intimately associated with viral infection. Moreover, WGCNA analysis identified 243 genes firmly associated with the pathological process of COVID-19. Subsequently, 50 common genes were screened between COVID-19 and FM, and functional enrichment analysis of these common genes primarily involved in immunerelated pathways. Among these common genes, 3 key genes were recognised by machine learning for the diagnosis of COVID-19 related FM. We also developed a diagnostic nomogram to predict the risk of FM occurrence which showed excellent predictive performance. Finally, we found that these 3 key genes were closely relevant to immune cells and screened potential drugs that interacted with the key genes. CONCLUSIONS: Our study revealed the bridge role of immune dysregulation between COVID-19 and fibromyalgia, and screened underlying biomarkers to provide new clues for further clinical research.

Identification of key biomarkers related to epithelial-mesenchymal transition and immune infiltration in ameloblastoma using integrated bioinformatics analysis.

OBJECTIVE: This study aimed to elucidate the underlying mechanisms of ameloblastoma (AM) through integrated bioinformatics analysis. METHODS: We downloaded two microarrays of AMs from the GEO database and identified differentially expressed genes (DEGs) by integrated bioinformatics analysis. The enrichment analysis of DEGs was conducted to characterize GO and KEGG pathways. Protein-protein interaction (PPI) network and hub genes were screened via STRING and Cytoscape. CIBERSORT algorithm was utilized to analyze immune infiltration in AMs. We also verified the diagnostic and therapeutic value of hub genes. RESULTS: Overall, 776 DEGs were identified in AMs through bioinformatics analysis. The function enrichment analysis shed light on pathways involved in AMs. Subsequently, we screened six hub genes via PPI network. Furthermore, we evaluated immune infiltration in AMs and found that macrophages may be participating in the progression of AMs. The upregulated expression of FN1 was related to the macrophages M2 polarization. Finally, ROC analysis indicated that six hub genes had high diagnostic value for AMs and 11 drugs interacted with upregulated hub genes were identified by screening the DGIdb database. CONCLUSION: This study revealed the underlying mechanisms of pathogenesis and biological behavior of AMs and provided candidate targets for the diagnosis and treatment of AMs.

Customized three dimensional printed prosthesis as a novel intercalary reconstruction for resection of extremity bone tumours: a retrospective cohort study.

AIMS: The 3D-printed prosthesis (3DP) is a novel treatment for massive bone defect reconstruction after tumor resection. This study was aiming to explore the clinical efficacy of customized 3DP for intercalary reconstruction by comparing the clinical outcomes after implanting customized 3DP or conventional allograft in limb salvage surgery. METHODS: A total of 28 patients with extremity bone tumors who underwent customized 3DP or conventional allograft reconstruction between 2011 and 2018 at our institution were analyzed retrospectively. Among them, 14 cases received customized 3DP reconstruction (3DP group), and 14 cases received conventional allograft reconstruction (control group). Demographics, surgical outcomes, radiographical assessments, limb functions, and post-operative complications between these two groups were collected to evaluate clinical outcomes. RESULTS: No significant difference was observed in the demographics, mean intra-operative blood loss, MOSI scores, and MSTS scores between the two groups. Patients in 3DP group had a shorter operative time (157.9 vs 199.6 min, p = 0.03) and lesser number of fluoroscopy (4.1 vs 8.1, p < 0.001) compared to control group. The mean time to osseointegration at bone-implant interfaces in 3DP group was significantly earlier than that in control group (6.1 vs 12.2 months, p < 0.001). Moreover, the 3DP group had a significantly lower post-operative complication rate than the control group (7% vs 50%, p = 0.03). CONCLUSIONS: The customized 3DP might provide a promising strategy for intercalary reconstruction in limb salvage surgery with more precise reconstruction, higher surgical efficiency, and comparable satisfactory clinical outcomes.

Prognostic value of tumoral and peritumoral magnetic resonance parameters in osteosarcoma patients for monitoring chemotherapy response.

OBJECTIVES: To evaluate parameters of diffusion-weighted imaging (DWI) and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) as early imaging indicators of tumor histologic response to pre-operative neoadjuvant chemotherapy and as probable prognostic factors for event-free survival (EFS) and overall survival in osteosarcoma (OS) in both tumoral and peritumoral areas. METHODS: Thirty-four OS patients who received three courses of neoadjuvant chemotherapy followed by surgery during 2014-2018 were enrolled in this study. All patients underwent baseline and post-chemotherapy DWI and DCE-MRI. Lesion region was defined as the tumoral area and peritumoral area. Parameters of apparent diffusion coefficient, capacity transfer constant (Ktrans), elimination rate constant, extravascular extracellular space volume ratio (Ve), and initial area under the curve as well as corresponding differences between pre- and post-chemotherapy in lesion regions were evaluated. Receiver operating characteristic analysis was used to evaluate the diagnostic performance of these parameters. The associations of all parameters with tumor histologic response, EFS, and overall survival were also calculated. RESULTS: In the tumor area, moderate evidence was found that post-Ktrans was lower in responders as compared with that in poor responders (p = 0.04, false discovery rate [FDR] corrected), and ΔKtrans exhibited significant between-groups differences (p = 0.04, Bonferroni corrected; or p = 0.006, FDR corrected). Weak evidence for the between-groups difference was found in the Ve in the peritumoral area (p = 0.025 before treatment and p = 0.021 after treatment, uncorrected). Furthermore, lower post-Ktrans in the tumoral area and lower pre-Ve in the peritumoral area were significant prognostic indicators for longer EFS (p = 0.002, p = 0.026) and overall survival (p = 0.003, p = 0.023). CONCLUSIONS: In OS, DWI and DCE-MRI parameters in both tumoral and peritumoral areas can reflect the chemotherapy response and prognosticate EFS and overall survival. KEY POINTS: • Peritumoral MRI parameters can reflect the chemotherapy response in OS patients. • Peritumoral MRI parameters can predict EFS and overall survival in OS patients. • MRI parameters may be predictive factors for evaluating chemotherapy efficacy and EFS.

Decentralized Privacy-Preserving Data Aggregation Scheme for Smart Grid Based on Blockchain.

As a next-generation power system, the smart grid can implement fine-grained smart metering data collection to optimize energy utilization. Smart meters face serious security challenges, such as a trusted third party or a trusted authority being attacked, which leads to the disclosure of user privacy. Blockchain provides a viable solution that can use its key technologies to solve this problem. Blockchain is a new type of decentralized protocol that does not require a trusted third party or a central authority. Therefore, this paper proposes a decentralized privacy-preserving data aggregation (DPPDA) scheme for smart grid based on blockchain. In this scheme, the leader election algorithm is used to select a smart meter in the residential area as a mining node to build a block. The node adopts Paillier cryptosystem algorithm to aggregate the user's power consumption data. Boneh-Lynn-Shacham short signature and SHA-256 function are applied to ensure the confidentiality and integrity of user data, which is convenient for billing and power regulation. The scheme protects user privacy data while achieving decentralization, without relying on TTP or CA. Security analysis shows that our scheme meets the security and privacy requirements of smart grid data aggregation. The experimental results show that this scheme is more efficient than existing competing schemes in terms of computation and communication overhead.

TLR4-RelA-miR-30a signal pathway regulates Th17 differentiation during experimental autoimmune encephalomyelitis development.

BACKGROUND: Toll-like receptor 4 (TLR4) is well known for activating the innate immune system; however, it is also highly expressed in adaptive immune cells, such as CD4+ T-helper 17 (Th17) cells, which play a key role in multiple sclerosis (MS) pathology. However, the function and governing mechanism of TLR4 in Th17 remain unclear. METHODS: The changes of TLR4 in CD4+ T cells from MS patients and experimental autoimmune encephalomyelitis (EAE) mice were tested. TLR4-deficient (TLR4-/-) naïve T cells were induced in vitro and transferred into Rag1-/- mice to measure Th17 differentiation and EAE pathology. DNA sequence analyses combining with deletion fragments and mutation analyses, chromatin immunoprecipitation (ChIP), and electrophoretic mobility shift assay (EMSA) were used to explore the mechanism of TLR4 signaling pathway in regulating Th17 differentiation. RESULTS: The levels of TLR4 were increased in CD4+ Th17 cells both from MS patients and EAE mice, as well as during Th17 differentiation in vitro. TLR4-/- CD4+ naïve T cells inhibited their differentiation into Th17, and transfer of TLR4-/- CD4+ naïve T cells into Rag1-/- mice was defective in promoting EAE, characterized by less demyelination and Th17 infiltration in the spinal cord. TLR4 signal enhanced Th17 differentiation by activating RelA, downregulating the expression of miR-30a, a negative regulator of Th17 differentiation. Inhibition of RelA activity increased miR-30a level, but decreased Th17 differentiation rate. Furthermore, RelA directly regulated the expression of miR-30a via specific binding to a conserved element of miR-30a gene. CONCLUSIONS: TLR4-/- CD4+ naïve T cells are inadequate in differentiating to Th17 cells both in vitro and in vivo. TLR4-RelA-miR-30a signal pathway regulates Th17 differentiation via direct binding of RelA to the regulatory element of miR-30a gene. Our results indicate modulating TLR4-RelA-miR-30a signal in Th17 may be a therapeutic target for Th17-mediated neurodegeneration in neuroinflammatory diseases.

Quantitative profiling of neurotransmitter abnormalities in brain, cerebrospinal fluid, and serum of experimental diabetic encephalopathy male rat.

Diabetic encephalopathy (DE), one of the most prevalent chronic complications of diabetes mellitus, is short of effective prevention and formidable therapeutic strategies. The aim of the present study is to reveal the imbalance of tryptophan (Trp) and its metabolites in streptozotocin (STZ)-induced experimental DE rats to underscore their critical values in clinical diagnosis of the disease. For this purpose, we first developed an accurate and appropriate simultaneous method for measuring Trp and its metabolites using liquid chromatography-tandem mass spectrometry, which was in accordance with the requirements of biological sample analysis. Secondly, a single STZ intraperitoneal injection was administered to male Sprague-Dawley rats, and their cognitive function was detected by Morris water maze tests. Cerebrospinal fluid (CSF), serum, and brain tissue were then collected for the determination of Trp and its metabolites. Compared with age-matched control rats, the levels of neuroprotective serotonin decreased significantly in the samples of cortices, hippocampi, striatum, CSF, and serums in the STZ-induced DE rats, while the levels of neurotoxic 3-hydroxykynurenine increased significantly. Moreover, analogous changes of both compounds were found in the central nervous system and peripheral blood of the STZ-induced DE rats. In conclusion, we established a quantitative method for the simultaneous detection of Trp and its metabolites, and we also present a critical elucidation of the nervous system dysfunction in DE.

miR-219 attenuates demyelination in cuprizone-induced demyelinated mice by regulating monocarboxylate transporter 1.

Remyelination is limited in patients with multiple sclerosis (MS) due to the difficulties in recruiting proliferating oligodendrocyte precursors (OPCs), the inhibition of OPC differentiation and/or maturation, and/or failure in the generation of the myelin sheath. In vitro studies have revealed that miR-219 is necessary for OPC differentiation and monocarboxylate transporter 1 (MCT1) plays a vital role in oligodendrocyte maturation and myelin synthesis. Herein, we hypothesized that miR-219 might promote oligodendrocyte differentiation and attenuate demyelination in a cuprizone (CPZ)-induced demyelinated model by regulating the expression of MCT1. We found that CPZ-treated mice exhibited significantly increased anxiety in the open field test. However, miR-219 reduced anxiety as shown by an increase in the total distance, the central distance and the mean amount of time spent in the central area. miR-219 decreased the quantity of OPCs and increased the number of oligodendrocytes and the level of myelin basic protein (MBP) and cyclic nucleotide 3' phosphodiesterase (CNP) protein. Ultrastructural studies further confirmed that the extent of demyelination was attenuated by miR-219 overexpression. Meanwhile, miR-219 also greatly enhanced MCT1 expression via suppression of oligodendrocyte differentiation inhibitors, Sox6 and Hes5, treatment with the MCT1 inhibitor α-cyano-4-hydroxycinnamate (4-CIN) reduced the number of oligodendrocytes and the protein levels of MBP and CNP. Taken together, these results suggest a novel mode of action of miR-219 via MCT1 in vivo and may provide a new potential remyelination therapeutic target.

Quercetin ameliorates learning and memory via the Nrf2-ARE signaling pathway in d-galactose-induced neurotoxicity in mice.

Aging is accompanied by deficits in cognitive function and neuronal degeneration or loss. Quercetin is a flavonoid that exhibits powerful antioxidant activity. This study evaluated the protective effects and mechanisms of quercetin in d-galactose-induced neurotoxicity in mice. Quercetin was administered daily at doses of 20 or 50 mg/kg in d-galactose-injected (50 mg/kg/subcutaneous (s.c.)) mice for eight weeks. Morris water maze tests demonstrated that quercetin significantly improved learning and memory compared to d-galactose-treated control mice. Quercetin also prevented changes in the neuronal cell morphology and apoptosis in the hippocampus as well as increased the expression of Nrf2, HO-1 and SOD in d-galactose-treated mice. Treatment with the Nrf2 inhibitor Brusatol reversed the effects of quercetin on HO-1 and SOD expression as well as neuronal cell protection. In conclusion, quercetin protected mice from d-galactose-induced cognitive functional impairment and neuronal cell apoptosis via activation of the Nrf2-ARE signaling pathway.

MiR-30a inhibits Th17 differentiation and demyelination of EAE mice by targeting the IL-21R.

T helper cells 17 (Th17) are recognized as key participants in the pathogenesis of chronic autoimmune diseases such as multiple sclerosis (MS). Regulation of Th17 differentiation is a valuable strategy for diagnosis and treatment of these complicated immune disorders. Here, by genome-wide expression profiling of microRNAs (miRs), we screened miR-30a, whose level was greatly decreased during Th17 differentiation and the process of demyelination disease, both in MS patients and experimental autoimmune encephalomyelitis (EAE) mice. Enforced constitutive expression of miR-30a in naïve T cells inhibited their differentiation into Th17, and in vivo overexpression of miR-30a resulted in fewer Th17 and alleviative EAE. Moreover, target prediction analysis and dual luciferase report assay revealed that interleukin-21 receptor (IL-21R) was a direct target of miR-30a, a finding consistent with the results that miR-30a downregulated the expression of IL-21R, while overexpression of IL-21R alleviated the inhibitory effect of miR-30a on Th17 differentiation. Taken together, our findings imply that miR-30a inhibits Th17 differentiation and the pathogenesis of MS by targeting IL-21R.

[Anti-oxidantive effect of methylene blue on acute lung injury induced by paraquat in rat].

OBJECTIVE: To study the protection effects of methylene blue on acute lung injury that induced by paraquat in rats. METHODS: 24 male SD rats were randomly distributed into four groups of 6 rats each group including control group, methylene blue group, paraquat group and paraquat plus methylene blue group. The rats were challenged acutely with paraquat( 25 mg/kg, i. p. ). After two hours treatment, the rats in paraquat plus methylene group were treated by methylene blue( 2 mg/kg, i. p. ) or vehicle treatment for 24 hours. Then, lung specimens were collected for pathological examinations and biochemical analysis including content of malondialdehyde( MDA) and activity of lactate dehydrogenase( LDH) and superoxide dismutase( SOD). The bronchoalveolar lavage fluids were also collected for the analysis of myeloperoxidase( MPO) activity. RESULTS: Compared to control group, the rats treated with paraquat induced acute lunginjury which was reflected by the increase of pathological score( P < 0. 05) and wet/dry ratio of lung tissue( P < 0. 05). Moreover, the content of MDA and LDH were increased, but the activity of SOD was decreased in lung tissues, and the activity of MPO were increased in bronchoalveolar lavage fluid in rats treated with paraquat. Compared to the rats with paraquat exposure, the pathological score( P < 0. 05) and wet/dry ratio( P <0. 05) in the rats treated with paraquat plus methylene blue were decreased. The content of MDA( P < 0. 05) and LDH( P < 0. 05) were also decreased, but the activity of SOD was increased( P < 0. 05). Moreover, the activity of MPO in bronchoalveolar lavage fluid was decreased in paraquat exposure rats with methylene blue treatment. CONCLUSION: Acute lung injury that induced by paraquat in rats is protected by methylene blue. The result may provide a new way to treat patients with acute paraquat poisoning.

Individualized chemotherapy for osteosarcoma and identification of gene mutations in osteosarcoma.

The study aims to identify novel gene mutations in osteosarcoma and to guide individualized preoperative chemotherapy for osteosarcoma based on the analysis of expression and mutations of the drug-metabolism-related genes. Twenty-eight osteosarcoma patients received individualized preoperative chemotherapy regimens. Expression levels and mutations of chemotherapy-related genes in samples collected from the patients were determined using real-time PCR and DNA sequencing, respectively. Patient sensitivity to chemotherapeutic agents was evaluated by systematic analysis of the PCR and sequencing results. Novel mutations were identified via high-throughput sequencing of 339 genes in 10 osteosarcoma samples. Individualized preoperative chemotherapy outcomes were valid for nine patients (n = 9/28, 32.1%). Chemosensitivity assays showed that all 28 patients were sensitive to ifosfamide, whereas 46.4 and 39.2% were sensitive to docetaxel and platinum, respectively. More importantly, patients receiving highly chemosensitive chemotherapy agents had better prognosis and treatment outcomes than those receiving less chemosensitive agents (P < 0.05). In addition, 39 gene mutations were detected in at least five osteosarcoma tumor samples. Analysis of the expression and mutation of drug-metabolism-related genes will aid in the design of effective individualized preoperative chemotherapy regimens for osteosarcoma. Determining the chemosensitivity of individual tumors to chemotherapeutic agents will facilitate the development of better therapeutic approaches. Individualized treatment of osteosarcoma may improve chemotherapy efficacy and the survival rate of osteosarcoma patients. High-throughput genotyping allows mapping of osteosarcoma mutations, and novel gene mutations offered new candidates for diagnosis and therapeutic targeting.

Quantitative-profiling of neurotransmitter abnormalities in the disease progression of experimental diabetic encephalopathy rat.

Diabetic encephalopathy (DE) is one of the most prevalent chronic complications of diabetes mellitus (DM), with neither effective prevention nor proven therapeutic regimen. This study aims to uncover the potential dysregulation pattern of the neurotransmitters in a rat model of streptozotocin (STZ)-induced experimental DE. For that purpose, male Sprague-Dawley (SD) rats were treated with a single intraperitoneal injection of STZ. Cognitive performance was detected with the Morris water maze (MWM) test. Serum, cerebrospinal fluid (CSF), and brain tissues were collected to measure the levels of neurotransmitters. Compared with the control rats, the acetylcholine (ACh) levels in serum, CSF, hippocampus, and cortex were all significantly down-regulated as early as 6 weeks in the STZ treatment group. In contrast, the glutamate (Glu) levels were decreased in CSF and the hippocampus, but unaffected in the serum and cortex of STZ-treated rats. As for γ-aminobutyric acid (GABA), it was down-regulated in serum, but up-regulated in CSF, hippocampus, and the cortex in the STZ-treated group. The mRNA expressions of neurotransmitter-related rate limiting enzymes (including AChE, GAD1, and GAD2) and pro-inflammatory cytokines (including IL-1ß and TNF-α) were all increased in the DE rats. Our data suggest that DM induces isoform-dependent and tissue-specific neurotransmitter abnormalities, and that neuroinflammation may underlay the nervous system dysfunction observed in the progression of DE.

Does Microwave Ablation of the Tumor Edge Allow for Joint-sparing Surgery in Patients With Osteosarcoma of the Proximal Tibia?

BACKGROUND: Joint-sparing surgery of a patient's native joint for osteosarcoma likely affords better function and comparable survival. However, it sometimes is challenging to resect a juxtaarticular osteosarcoma in a way that preserves the affected epiphysis because wide margins are necessary to minimize the risk of local recurrence. If there was a method to resect a tumor close to the joint and treat a potentially positive margin to prevent recurrence, it might allow salvage of a joint that otherwise might be lost. QUESTIONS/PURPOSES: We therefore asked (1) whether joint-preserving tumor resection could be performed for juxtaarticular osteosarcoma after microwave ablation of the tumor edge under navigation without leading to local recurrences, (2) what is the resulting function, and (3) what are the complications associated with this procedure. METHODS: Between 2009 and 2011, we treated 11 patients who had juxtaarticular osteosarcoma of the proximal tibia (mean age, 12 years; range, 9-16 years) with joint-preserving surgery by transepiphysis tumor resection after navigation-assisted microwave ablation of the tumor edge; they were followed a minimum of 37 months (mean, 48 months; range 37-62 months), and none was lost to followup. Patients were considered eligible for this procedure if they had a distance from the tumor edge to the articular surface between 10 to 15 mm, good chemotherapy responses, no pathologic fracture and no tumor involvement of major neurovascular structures. Allograft in combination with a vascularized fibula flap was used for segmental reconstruction. We recorded local tumor control, complications, and functional outcomes using the Musculoskeletal Tumor Society score, which ranges from 0 to 30, with higher scores indicating better function. RESULTS: There were no local recurrences. Major complications included osteonecrosis of part of the epiphysis in two patients and deep infection in one. The Musculoskeletal Tumor Society score ranged from 26 to 30 with a mean of 29. CONCLUSIONS: In selected patients with osteosarcoma invading the epiphysis, navigated resection facilitates performing joint-sparing surgery, and in our small series, the adjuvant microwave ablation seemed to provide adequate local tumor control. Although more experience and longer followup are needed, this approach may make it possible to salvage more native joints when performing limb salvage for osteosarcoma. LEVEL OF EVIDENCE: Level IV, therapeutic study.

Giant cell tumor of axial vertebra: surgical experience of five cases and a review of the literature.

BACKGROUND: Due to the complex anatomy of the upper cervical spinal column region and the variable aggressiveness of giant cell tumors (GCTs), there exists no standard treatment for GCTs of axial vertebra. To the best of our knowledge, there are only a few case reports in the literature and no large sum numbers of clinical trials about the treatment of, or research into, axial vertebra GCTs. METHODS: Between 2009 and 2013, five patients pathologically diagnosed with axial vertebra GCTs were treated at our hospital. We performed intralesional excision and odontoid process reconstructive surgery to preserve the odontoid process, followed with adjuvant radiation therapy after surgery. RESULTS: For those with an intact bone shell, part of the ß-TCP (beta tricalcium phosphate) artificial bone could be seen clearly after surgery and became blurred three months after surgery, as seen on a radiograph. One year later, the part of ß-TCP artificial bone was fused as a block. Subsequently, autogenous bone regenerated successfully and artificial bone degraded thoroughly. For those with a defective cortical bone, partial fusion of the odontoid process, autograft ilium and third vertebra body could be seen three months after surgery, and complete fusion was seen nine months later. The odontoid process was preserved successfully, and the upper cervical spine was reconstructed effectively, without implant failure or infection. CONCLUSIONS: In this study, the odontoid process and function of upper cervical vertebra was preserved successfully through lesion curettage, combined with reconstruction with bone grafting, and adjuvant radiation therapy after surgery. During the follow-up periods, no recurrence or complications was observed.