Endothelial cell-specific molecule-1 as an invasiveness marker for pituitary null cell adenoma.

BACKGROUND: Endothelial cell-specific molecule-1 (ESM-1) is a biomarker associated with tumor progression in pituitary adenoma. We specifically focused on one type of pituitary adenoma, namely null cell adenoma (NCA) and evaluated the relationship between invasion and ESM-1 expression in both vascular endothelial and adenoma tissues. METHODS: Tissue samples from 94 patients with pituitary NCA were obtained through microscopic transsphenoidal resection. Tumor size and invasion were determined through preoperative magnetic resonance imaging. Immunohistochemical staining was performed to detect ESM-1 expression. ESM-1 index of ≥3 was defined as high expression. RESULTS: Signs of invasion were observed in 46 (47.9%) of the 94 patients. Significant differences were observed in the invasion state and maximum tumor diameter between high and low expression of ESM-1 in vascular endothelial tissues (both P < 0.05). Significant positive associations were noted between ESM-1 expression in vascular endothelial tissues and tumor invasion (P = 0.002) and tumor size (P = 0.020). However, only tumor size was associated with ESM-1 expression in adenoma tissues (P = 0.016). CONCLUSION: In NCA, a significant positive association between tumor invasion and ESM-1 expression was observed only in vascular endothelial tissues, suggesting that tumor progression occurs mainly through ESM-1-associated mechanism.

The Influence of Supervisor-Postgraduate Relationship on Master's Students' Research Learning Engagement-The Mediating Effect of Academic Aspiration.

Research learning engagement is the basic element of master's students' innovation output, and the supervisor is the first responsible body for master's students' cultivation. Exploring the influence of the supervisor-postgraduate relationship on master's students' research learning engagement, with a focus on the mediating role of academic aspiration, is of great significance for the improvement of master's students' cultivation quality. We surveyed 569 master's students at a university in Wuhan, China, using 3 measurement tools: the Supervisor-Postgraduate Relationship Scale, the Research Learning Engagement Scale, and the Academic Aspirations Scale. The results showed that: (1) The supervisor-postgraduate relationship positively and significantly predicted master's students' research learning engagement, and academic aspiration played a fully mediating role in the process. (2) There were differences in the effects of the three dimensions of the supervisor-postgraduate relationship on master's students' research learning engagement, with research collaboration having the greatest total effect on the impact of master's students' research learning engagement. This study emphasizes the importance of the supervisor-postgraduate relationship and academic aspirations and provides some implications for improving the research learning engagement of master's students.

MRI 3D SPACE T2WI for Pituitary Adenoma Cavernous Sinus Invasion Diagnosis.

OBJECTIVE: This study aims to assess the utility of magnetic resonance imaging (MRI) 3D SPACE T2-weighted imaging (T2WI) sequences in evaluating cavernous sinus invasion by pituitary adenomas. METHODS: Data were collected from patients who underwent continuous pituitary MRIexaminations at the Medical Imaging Center of our hospital from October 2019 to February 2021. Eligible cases were evaluated for sagittal and axial T1WI sequences, coronal 3D SPACE T2WI sequences, and sagittal and coronal enhanced T1-weighted imaging (T1WI) sequences using the INFINITT PACS workstation. The Wilcoxon signed-rank test for paired samples and the Mann-Whitney U test for 2 independent samples were used to statistically analyze differences in image quality scores among various groups. In addition, the sensitivity, specificity, positive predictive value, and negative predictive value of each observation index were compared with intraoperative results. RESULTS: 3D SPACE T2WI showed superior cavernous sinus imaging quality compared with contrast enhanced T1WI and T2WI plain scans (P < 0.05). The sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 90.0%, 55.60%, and 100.0%, respectively. The accuracy for pituitary adenoma invasiveness diagnosis based on cavernous sinus medial wall integrity was 94.40%. CONCLUSIONS: The imaging quality of the medial wall of the cavernous sinus on the 3D SPACE T2WI plain scan sequence surpassed that of contrast enhanced T1WI TSE-enhanced scans and T2WI TSE plain scans. The continuous observation of the medial wall of the cavernous sinus using this sequence holds great diagnostic value for assessing cavernous sinus invasion by pituitary adenomas. This strategy is more reliable than traditional MRI observation indicators.

Analysis of synthetic cannabinoids in wastewater of major cities in China.

Synthetic cannabinoids (SCs) could pose serious health risks to its users. It is necessary to monitor its community consumption. Wastewater-based epidemiology is a potentially useful approach in this regard. However, limited research has been conducted to investigate the occurrence of SCs in wastewater. In this study, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was optimized to analyze 8 SCs and metabolites (in total 16 analytes) in wastewater. The limit of quantification for this method for certain analytes in wastewater was as low as 0.03 ng L-1. The validated method was used to examine the stability of the analytes under different conditions and to examine their occurrence in wastewater collected from 31 major cities across China. The overwhelming majority of the analytes were stable within 24 h, even at room temperature. However, 5-fluoro MDMB-PICA and MDMB-4en-PINACA butanoic acid metabolite showed significant degradation within 120 days even when stored at -20 °C or -80 °C. At least one cannabinoid or their metabolite was detected in 21 cities. In the city with the highest detection rate, at least one synthetic cannabinoid or metabolite was detected in 95% of samples of the city. MDMB-4en-PINACA butanoic acid metabolite had the highest detection frequency (in 13.4% of the samples). These results indicated that SCs were used in a significant number of Chinese cities. A few parent drugs (MDMB-4en-PINACA, ADB-BUTINACA, 5-fluoro MDMB-PICA, 4-fluoro MDMB-BUTINACA) were detected in a small fraction of wastewater samples, possibly due to release from manufacturing of these cannabinoids or illegal addition of electronic cigarettes.

The "Double-Peak" Pattern of Pituitary Adenoma Intrasellar Pressure and Its Effects on the Microvascular Structure.

OBJECTIVE: The purpose of the present study was to investigate the relationship between the intrasellar pressure (ISP) and the microvascular structure of pituitary adenomas. METHODS: We retrospectively analyzed the ISP in 66 patients with pituitary adenomas. The corresponding microvascular structure was obtained using immunohistochemistry and analyzed for its correlation with the ISP. RESULTS: The average ISP was 25.89 ± 8.27 mm Hg, and the ISP was not related to the size of the adenoma (Pearson correlation coefficient, 0.103; P = 0.415). The ISPs of adenomas with different Knosp grades were significantly different (P < 0.05). From grade 0 to grade 4, at first, the ISP increased with the Knosp grade and reached the first peak at grade 2. It then decreased at grade 3 and increased again at grade 4, showing a "double-peak" pattern. The minimal diameter and perimeter of the microvessels and the vessel-covered area percentage were positively related to the ISP. When these parameters were compared among the adenomas of different Knosp grades, they also exhibited a "double-peak" pattern. CONCLUSIONS: In the present study, we found that with the increase in pituitary adenoma size and invasion of the surrounding tissues, the ISP of pituitary adenomas showed a "double-peak" pattern. The ISP and certain parameters of the microvascular structure are related, because the microvasculature adaptively changes its structure in response to the changing ISP to ensure a sufficient blood supply to the adenoma. The specific mechanism of this phenomenon requires further study.

MRI and pathological features of Rathke cleft cysts in the sellar region.

The aim of the present study was to investigate the MRI and pathological features of Rathke cleft cysts (RCC) in the sellar region. A total of 45 RCC cases were retrospectively analyzed. RCC size, location, intracyst nodules and general signals, as well as the posterior pituitary bright spot (PPBS) were analyzed using MRI-T1 weighted images (T1WI) and T2WI. The relationship between the presence of PPBS and histopathological features was additionally evaluated. On T1WI, there were 18 cases of isointense signal, 16 cases of hyperintense signal, 9 cases of hypointense signal, 1 case of heterogeneous signal and 1 case with a stratification effect, with isointense signal in the upper part and hyperintense signal in the lower part. On T2WI, there were 5 cases of isointense signal, 27 cases of hyperintense signal, 11 cases of hypointense signal and 1 case of the stratification effect. There were 10 cases of PPBS+ and 35 cases of PPBS-. There were no significant differences in the age, sex, cyst location and size between PPBS+ and PPBS- cases. However, PPBS+ cases had significantly lower inflammation than PPBS- cases. A total of 20 cases of intracystic nodules were identified on MRI scans, most of which exhibited T2 -hypointense signals. The shape of RCC nodules varied and there were 17 cases where the nodules were non-adherent to the cyst wall. The MRI signals of RCCs varied and most nodules were floating within cysts. Intracystic nodules are characteristic features of RCCs when observed by MRI and thus are of high diagnostic value. Most patients with RCC were also PPBS-, which may be associated with an increased inflammatory response.

A Novel Scoring System Based on Preoperative Routine Blood Test in Predicting Prognosis of Atypical Meningioma.

PURPOSE: The aim of this study was to explore the correlation and clinical significance of preoperative fibrinogen and neutrophil-lymphocyte ratio (F-NLR) scoring system with 3-year progression-free survival (PFS) of patients with atypical meningioma. MATERIALS AND METHODS: Clinical, pathological, radiological, and laboratory variables were collected to analyze their correlation with 3-year PFS in the training set with 163 patients. Patients were classified by different F-NLR scores (0, 1, or 2). External validation for the predictive value of F-NLR scoring system was performed in the validation set with 105 patients. RESULTS: Overall, 37.3% (100 of 268) of the enrolled patients were male. The scoring system showed good performance in predicting 3-year PFS (AUC = 0.872, 95%CI = 0.811-0.919, sensitivity = 66.1%, specificity = 93.3%, and Youden index = 0.594). DeLong's test indicated that the AUC of F-NLR scoring system was significantly greater than that of fibrinogen level and NLR (Z = 2.929, P = 0.003; Z = 3.376, P < 0.001). Multivariate Cox analysis revealed that tumor size (HR = 1.39, 95%CI = 1.10-1.76, P = 0.007), tumor location (HR = 3.11, 95%CI = 1.60-6.95, P = 0.001), and F-NLR score (score of 1: HR = 12.78, 95%CI = 3.78-43.08, P < 0.001; score of 2: HR = 44.58, 95%CI = 13.02-152.65, P < 0.001) remained significantly associated with 3-year PFS. The good predictive performance of F-NLR scoring system was also demonstrated in the validation set (AUC = 0.824, 95%CI = 0.738-0.891, sensitivity = 62.5%, specificity = 87.9%, and Youden index = 0.504). CONCLUSION: Our study confirmed the correlation and clinical significance of preoperative F-NLR scoring system with 3-year PFS of patients with atypical meningioma. A prospective and large-scale study is required to validate our findings.

Application of the Three Points and Three Lines Method to Accurately Open the Sellar Floor in Microscopic Transsphenoidal Surgery of Pituitary Adenomas.

OBJECTIVE: In this study, the sellar floor morphology of patients with pituitary adenoma is analyzed and a simple yet reliable method is identified to determine the location of bone window opening. METHODS: Clinical information of 144 consecutively admitted patients was retrospectively analyzed. Enhanced magnetic resonance imaging of the midsagittal plane was selected as the reference for classifying the sellar floor. Intraoperative tumor location, extent of tumor resection, and follow-up results were analyzed for different types of sellar floor. The tuberculum sellae, lowest point of the sphenoid sinus, and the lowest point of the sellar floor and 3 lines related to them were used to classify the sellar floor. This is referred to as the "three points and three lines" method. RESULTS: Based on its location in the sphenoid sinus, the sellar floor can be classified into 4 types: 12 patients (8.3%) with high sellar, 70 (48.6%) with medium sellar, 30 (20.8%) with low sellar, and 32 (22.8%) with steep sellar. The maximum tumor diameter, maximum sellar floor diameter, and the intercarotid distance were all significantly different among patients with different types of sellar floor (P < 0.001). For all patients, quick intraoperative location of the sellar floor opening was achieved. A total of 104 patients (72.2%) had total tumor resection, 28 (19.40%) had subtotal tumor resection, and 4 (2.8%) had partial tumor resection. Twenty patients (13.9%) experienced cerebrospinal fluid leak, and there was no significant difference in cerebrospinal fluid leak rate among groups. CONCLUSIONS: Presurgical classification and location of the sellar floor are critical for understanding and assessing the transsphenoidal approach. Different types of sellar floor appeared in the surgery with different morphologic features. The three points and three lines method helps the surgeon to predetermine the location of the sellar floor opening and to shorten surgical time.

Effect of HMGB1 and RAGE on brain injury and the protective mechanism of glycyrrhizin in intracranial­sinus occlusion followed by mechanical thrombectomy recanalization.

The key to successful treatment of cerebral venous­sinus occlusion (CVO) is the rapid recanalization of the sinus following venous­sinus occlusion; however, rapid recanalization of the sinus may also cause secondary cerebral injury. The present study examined mechanical thrombectomy­related brain injury and the possible molecular mechanisms following CVO recanalization, and investigated the protective effect of glycyrrhizin (GL) in CVO recanalization. The cerebral venous sinus thrombosis (CVST) model was induced in rats using 40% FeCl3. Mechanical thrombectomy was performed at 6 h post­thrombosis. GL was administered to rats following thromboembolism. Neurological function and brain water content were measured prior to sacrifice of the rats. Serum malondialdehyde, superoxide dismutase and nitric­oxide synthase concentrations were measured. The expression levels of high­mobility group box 1 (HMGB1) and receptor of advanced glycation end products (RAGE) and its downstream inflammatory mediators were measured in serum and brain tissues. Rapid CVO recanalization caused brain injury, and the brain parenchymal damage and neurological deficits caused by CVO were not completely restored following recanalization. Similarly, following rapid recanalization in the venous sinus, the expression levels of HMGB1 and RAGE were lower than those in the CVST group, but remained significantly higher than those of the sham group. The combination of mechanical thrombectomy and GL improved cerebral infarction and cerebral edema in rats, and inhibited the extracellular transport of HMGB1, and the expression of downstream inflammatory factors and oxidative­stress products. The administration of exogenous recombinant HMGB1 reversed the neural protective effects of GL. In conclusion, mechanical thrombectomy subsequent to CVO in rats can cause brain injury following recanalization. HMGB1 and RAGE promote inflammation in the process of brain injury following recanalization. GL has a relatively reliable neuroprotective effect on brain injury by inhibiting HMGB1 and its downstream inflammatory factors, and decreasing oxidative stress.

Poor Brain-Tumor Interface-Related Edema Generation and Cerebral Venous Decompensation in Parasagittal Meningiomas.

BACKGROUND: Parasagittal meningioma (PSM) has a high incidence of peritumoral edema and unclear pathogenesis. The venous compression theory has been proposed as a pathomechanism; however, this is controversial, and the various edema patterns have not been recognized. OBJECTIVE: We sought to establish the relationship between venous circulation status with different edema patterns in PSM and the neurologic outcomes of these different patterns. METHODS: We performed a retrospective study of 60 consecutive patients who underwent surgical treatment for PSM. Patients were divided into 3 groups: no edema, poor brain-tumor interface-related edema (PIRE), and strong brain-tumor interface-related edema (SIRE). Single-blinded observers scored venous circulation for each patient based on the degree of superior sagittal sinus (SSS) occlusion, the number of involved cortical veins, and venous collateral grade. PIRE and SIRE were analyzed using multivariate analysis. Finally, we evaluated the functional independence and mobility score for every patient. RESULTS: The PIRE group showed the highest rate of cerebral venous decompensation at 75% (n = 15) compared with 38.5% (n = 5) in the SIRE group and 22.2% (n = 6) in the no-edema group. We observed a significant correlation between venous decompensation and PIRE generation on multivariate analysis (P = 0.029). The PIRE group showed the worst immediate functional status, and the SIRE group had the best improvement in complete dependence rate (23%) at late evaluation. CONCLUSIONS: The generation of PIRE, but not SIRE, may depend on venous decompensation in PSM. PIRE generation is predictive of worse neurologic outcome. Future studies into the pathogenesis of peritumoral edema should distinguish the different edema patterns.

Suppression of microRNA-130b inhibits glioma cell proliferation and invasion, and induces apoptosis by PTEN/AKT signaling.

Glioblastoma is the most common malignant brain tumor in adults and is characterized by extensive proliferation and the diffused invasion of tumor cells. Due to the intricate signaling pathways involved in glioma progression, more effective targeted therapies and prognostic biomarkers in clinical practice are required. The suppression of proto-oncogene function or recovery of tumor suppressor gene function remains one of the primary approaches in gene therapy. The close association between the abnormal expression or mutation of microRNA (miRNA) and the tumorigenesis, progression and staging in glioma have been demonstrated previously. However, the expression pattern and specific role of microRNA­130b (miR­130b) in the tumor occurrence and progression of glioma are unclear. In the present study, quantitative polymerase chain reaction was performed to determine the expression level of miR-130b in 30 brain glioma patients and 3 glioma cell lines. An miR­130b inhibitor was transfected into U87 cells to downregulate the expression of miR-130b, and assessments of cell proliferation, cell cycle, apoptosis, cell invasion and migration in vitro and nude mouse tumorigenicity in vivo were conducted. Western blotting and luciferase reporter gene technology were used to verify the downstream target gene of miR-130b, namely phosphatase and tensin homolog (PTEN). The results demonstrated that miR-130b expression was increased in glioma tissues and cell lines in comparison with non-glioma tissues or cells. The downregulated expression of miR-130b inhibited the proliferation and invasion of glioma cells, induced apoptosis of the cells in vitro and inhibited their tumorigenicity in vivo. Western blotting and luciferase reporter assays demonstrated that the PTEN gene is a direct target of miR­130b. Western blotting revealed that the miR-130b inhibitor upregulated the expression of PTEN, inhibited AKT pathway activation, upregulated the tumor suppressor gene p27, and suppressed cyclin D1, matrix metalloproteinase 2 and 9 expression. These results suggest that the miR-130b inhibitor suppressed glioma cell proliferation and invasion via the PTEN/AKT pathway. Therefore, miR­130b is suggested to be an effective therapeutic target for glioma.

GLUT3 expression in cystic change induced by hypoxia in pituitary adenomas.

Tumor cells require large amounts of energy to sustain growth. Through the mediated transport of glucose transporters, the uptake and utilization of glucose by tumor cells are significantly enhanced in the hypoxic microenvironment. Pituitary adenomas are benign tumors with high-energy metabolisms. We aimed to investigate the role of expression of glucose transporter 3 (GLUT3) and glucose transporter 1 (GLUT1) in pituitary adenomas, including effects on size, cystic change and hormone type. Pituitary adenomas from 203 patients were collected from January 2013 to April 2017, and immunohistochemical analysis was used to detect the expression of GLUT3 and GLUT1 in tumor specimens. GLUT3-positive expression in the cystic change group was higher than that in the non-cystic change group (P = 0.018). Proportions of GLUT3-positive staining of microadenomas, macroadenomas, and giant adenomas were 22.7 (5/22), 50.4 (66/131) and 54.0% (27/50), respectively (P = 0.022). In cases of prolactin adenoma, GLUT3-positive staining was predominant in cell membranes (P = 0.000006), while in cases of follicle-stimulating hormone or luteotropic hormone adenoma, we found mainly paranuclear dot-like GLUT3 staining (P = 0.025). In other hormonal adenomas, GLUT3 was only partially expressed, and the intensity of cell membrane or paranuclear punctate staining was weak. In contrast to GLUT3, GLUT1 expression was not associated with pituitary adenomas. Thus, our results indicate that the expression of GLUT3 in pituitary adenomas is closely related to cystic change and hormonal type. This study is the first to report a unique paranuclear dot-like GLUT3 staining pattern in pituitary adenomas.

Uneven Distribution of Regional Blood Supply Prompts the Cystic Change of Pituitary Adenoma.

BACKGROUND: Previous studies have suggested that the cystic change of pituitary adenoma might be related to the blood supply and metabolism of the tumor; however, the exact pathologic mechanism underlying the cystic change remains unknown. OBJECTIVES: We aimed to assess the features of regional blood supply of pituitary adenoma and examine its relationship with the cystic change of pituitary adenoma. METHODS: Patients (N = 79) with pituitary adenoma admitted to our hospital were divided into the parenchyma group (n = 40) or the cystic change group (n = 39). Dynamic contrast-enhanced magnetic resonance imaging of the pituitary adenoma was conducted for the parenchyma group and the steepest slopes (SSmax, reflecting regional blood supply) at different areas were calculated. The location of cystic change of the pituitary adenoma was recorded and analyzed for the cystic change group. RESULTS: The parenchyma group showed an upper SSmax of 2.52 ± 1.18, a lower SSmax of 2.89 ± 1.46, a left SSmax of 2.71 ± 1.31, and a right SSmax of 2.66 ± 1.29. The difference between the upper and lower SSmax was statistically significant (P < 0.001), with no difference between the left and right regions (P = 0.668). The location of cystic change of the pituitary adenoma was mainly in the upper region, accounting for 48.7% of cases. CONCLUSIONS: Regional blood supply is unevenly distributed in the parenchymal pituitary adenoma, with reduced blood supply in the upper than the lower region. Cystic change mainly occurs in the upper region of pituitary adenoma.

miR-130b regulates the proliferation, invasion and apoptosis of glioma cells via targeting of CYLD.

MicroRNAs are short non-coding RNAs that play important roles in gliomas. However, the role of miR-130b in glioma remains unclear. In the present study, miR-130b expression was upregulated in glioma tissues and cell lines. Kaplan-Meier analysis indicated that the upregulation of miR-130b expression correlated with poor prognoses in glioma patients. Multivariate analysis demonstrated that this upregulation and a high-grade classification were independent factors that both predicted poor outcomes for glioma patients. Dual-luciferase assays identified that the cylindromatosis (CYLD) gene is a direct target of miR-130b. Functional studies demonstrated that a miR-130b mimic significantly promoted the growth and invasion of glioma cells, while also inhibiting apoptosis via selective targeting of CYLD, which was enhanced by CYLD-targeted siRNA. In contrast, a miR­130b inhibitor suppressed these biological behaviors, and this inhibition was reversed by CYLD-targeted siRNA. These data revealed that miR-130b could act as a novel potential diagnostic biomarker for glioma, while also demonstrating the importance of miR­130b in the cell proliferation and progression of glioma, indicating that it may serve as a useful therapeutic target for glioma.

Analysis of Factors Related to Hypopituitarism in Patients with Nonsellar Intracranial Tumor.

OBJECTIVE: Previous studies have suggested that postoperative hypopituitarism in patients with nonsellar intracranial tumors is caused by traumatic surgery. However, with development of minimally invasive and precise neurosurgical techniques, the degree of injury to brain tissue has been reduced significantly, especially for parenchymal tumors. Therefore, understanding preexisting hypopituitarism and related risk factors can improve perioperative management for patients with nonsellar intracranial tumors. METHODS: Chart data were collected retrospectively from 83 patients with nonsellar intracranial tumors admitted to our hospital from May 2014 to April 2015. Pituitary function of each subject was determined based on results of preoperative serum pituitary hormone analysis. Univariate and multivariate logistic regression methods were used to analyze relationships between preoperative hypopituitarism and factors including age, sex, history of hypertension and secondary epilepsy, course of disease, tumor mass effect, site of tumor, intracranial pressure (ICP), cerebrospinal fluid content, and pituitary morphology. RESULTS: A total of 30 patients (36.14%) presented with preoperative hypopituitarism in either 1 axis or multiple axes; 23 (27.71%) were affected in 1 axis, and 7 (8.43%) were affected in multiple axes. Univariate analysis showed that risk factors for preoperative hypopituitarism in patients with a nonsellar intracranial tumor include an acute or subacute course (≤3 months), intracranial hypertension (ICP >200 mm H2O), and mass effect (P < 0.05). Multivariate logistic regression analysis showed that mass effect is an independent risk factor for preoperative hypopituitarism in patients with nonsellar intracranial tumors (P < 0.05; odds ratio, 3.197). CONCLUSIONS: Prevalence of hypopituitarism is high in patients with nonsellar intracranial tumors. The occurrence of hypopituitarism is correlated with factors including an acute or subacute course (≤3 months), intracranial hypertension (ICP >200 mm H2O), and mass effect (P < 0.05). Mass effect is an independent risk factor for hypopituitarism.

SDF-1 Expression is Associated with Poor Prognosis in Osteosarcoma.

BACKGROUND: Stromal cell-derived factor-1 (SDF-1) expression has been reported to be a predictor of poor clinical symptoms in certain types of cancer. Vascular endothelial growth factor (VEGF) is a well-known factor that mediates the micro-angiogenesis of solid tumors, and SDF-1 mediated expression of VEGF may promote tumor growth and metastasis, resulting in poor clinical outcome. Therefore, we explored the expression levels of SDF-1 and VEGF in patients with osteosarcoma in order to determine the association between their expression levels and unfavorable outcomes. METHODS: A total of 54 patients with osteosarcoma were included in the current study. The protein expression levels of SDF-1 and VEGF were evaluated on immunohistochemical and immunofluorescence staining. The correlation between the expression levels of SDF-1 and VEGF and their association with clinical parameters were analyzed using the Pearson chi-square test and the Spearman-rho test. Univariate and multivariate Cox regression analyses were used to identify potential prognostic factors. The Kaplan-Meier method was employed to analyze overall survival. RESULT: Low SDF-1 and VEGF expression levels were detected in 20.4% (11 of 54) and 22.2% (12 of 54) of the patients with osteosarcoma, respectively; moderate expression was detected in 35.2% (19 of 54) and 37.0% (20 of 54) of the patients, respectively; and high expression was detected in 44.4% (24 of 54) and 40.7% (22 of 54) of the patients, respectively. Protein levels of both SDF-1 and VEGF were significantly associated with the histologic grade (p=0.004 and p=0.042 respectively), the presence of metastasis (p=0.009 and p=0.028 respectively), and Enneking staging (p<0.001 and p=0.003 respectively). The association between expression levels of SDF-1and VEGF had a significantly positive correlation (p<0.001and r=0.618). The expression levels of both SDF-1 and VEGF were significantly associated with shorter overall survival on univariate analysis; however, the association was significant for SDF-1 expression alone in the multivariate analysis (p=0.26, hazard ratio =2.640 [1.124-6.200]). CONCLUSION: SDF-1 and VEGF expression levels were both significantly associated with osteosarcoma, and SDF-1 expression is a potential independent prognostic indicator in patients with osteosarcoma.

MicroRNA-218 modulates activities of glioma cells by targeting HMGB1.

To explore the effects of microRNA-218 (miR-218) on glioma cell lines and the related mechanism. U251 and U87 cells were transfected with negative control, miR-218 mimic or miR-218 inhibitor using lipofectamine 2000. The expressions of mRNA and proteins were detected with qRT-PCR and Western blotting. The cell proliferation, apoptosis, migration and invasion were studied using MTT, flow cytometry, Transwell assay and scratch-wound assay, respectively. The targeting effect of HMGB1 by miR-218 was measured with luciferase reporter assay. The results showed that miR-218 was significantly downregulated while HMGB1 was upregulated in both glioma cell lines. Transfection of miR-218 significantly reduced the cell viability and colony formation, increased cell apoptosis and arrested cell in G0/G1 phase. Transfection of miR-218 also decreased the invasion and migration of glioma cells. The expressions of HMGB1, RAGE, cyclin D1 and MMP-9 were downregulated while the expression of caspase-9 was upregulated by miR-218. Silencing HMGB1 increased the expression of RAGE, cyclin D1, MMP-9 but decreased the expression of caspase-9 in U251 and U87 cells. Co-transfection with pcHMGB1 and miR-218 significantly decreased the growth inhibition and increased the apoptosis of glioma cells while these effects were abolished in glioma cells co-transfected with HMGB1 siRNA and miR-218 inhibitor. In addition, co-transfection with pcHMGB1 and miR-218 inhibitor increased the invasiveness of U251 and U87 cells. These findings suggested that miR-218 may negatively regulate HMGB-mediated suppression of RAGE to regulate cell proliferation, apoptosis and invasion, and that intervention of miR-218-HMGB1-RAGE may be useful for developing potential clinical strategies.

MicroRNA-130b promotes cell proliferation and invasion by inhibiting peroxisome proliferator-activated receptor-γ in human glioma cells.

MicroRNA-130b (miR-130b) is a novel tumor-related miRNA that has been found to be involved in several biological processes. However, there is limited evidence regarding the role of miR-130b in the tumorigenesis of human gliomas. In the present study, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays were used to quantify miR-130b expression levels in human glioma tissues and glioma cell lines (U251, U87, SNB19 and LN229). The expression level of miR-130b was found to be markedly higher in human glioma tissues than in non­neoplastic brain specimens. Specifically, higher expression levels of miR­130b were observed in the glioma cell lines, compared with those in normal human astrocytes (NHA). We also confirmed that miR­130b interacted with the 3'-untranslated region of peroxisome proliferator­activated receptor-γ (PPAR­Î³), which negatively affected the protein levels of E-cadherin. Furthermore, its effects on cell proliferation and invasion were examined using CCK8, colony formation, cell cycle and Transwell assays. We found that the upregulation of miR-130b induced cell proliferation, decreased the percentage of cells in the G0/G1 phase and enhanced the invasiveness of U251 glioma cells whereas the downregulation of miR-130b exerted opposing effects. Moreover, it was demonstrated that the downregulation of miR­130b in U251 glioma cells restored the expression of PPAR-γ and E-cadherin, and inhibited the expression of ß-catenin. Notably, PPAR-γ knockdown abolished the inhibitory effect of miR-130b inhibitor on the proliferation and invasivness of U251 cells. Taken together, these findings suggest that miR­130b promotes the proliferation and invasion of U251 glioma cells by inhibiting PPAR-γ.

HIF-1α Silencing Inhibits the Growth of Osteosarcoma Cells by Inducing Apoptosis.

BACKGROUND: Osteosarcoma (OS) is a malignant tumor of mesenchymal origin, which is generally locally aggressive and tends to produce early systemic metastases. Therefore, an identification of a novel therapeutic target is required. METHODS: We investigated the effect of small interfering RNA (siRNA) targeting hypoxia-inducible factor 1α (HIF-1α) on the growth of Osteosarcoma MG-63 and U2-OS cells. Two cell-lines were transfected with various concentrations of HIF-1α or control siRNA, and the effect on HIF-1α expression was analyzed by using quantitative polymerase chain reaction and western blot analysis. The effects of HIF-1α siRNA on growth inhibition and apoptosis were then evaluated by standard methods. RESULT: HIF-1α siRNA treatment reduced HIF-1α mRNA and protein expression in MG-63 and U2-OS cells significantly. The downregulation of HIF-1α expression upregulated caspase-3 expression subsequently inhibited the growth of MG-63 and U2-OS cells and induced apoptosis of the two cells. Our in vitro data indicate that the downregulation of HIF-1α is capable of suppressing OS cells growth, through the induction of apoptosis. HIF-1α inhibition may be a new strategy for the treatment of Osteosarcoma.

TPX2 promotes glioma cell proliferation and invasion via activation of the AKT signaling pathway.

Glioblastoma multiforme (GBM) is the most common and most malignant type of primary adult brain cancer. The most common phenotype associated with GBM is cellular invasion; however, the molecular mechanisms governing this process are poorly understood. Targeting protein for Xenopus kinesin-like protein 2 (TPX2) is a nuclear protein with roles in cellular proliferation and mitotic spindle assembly. TPX2 is overexpressed in various malignancies, including human malignant astrocytoma. Despite this finding, the exact role of TPX2 in human glioma is not well defined. The present study reports the elevated expression of TPX2 in a number of glioma cell lines. TPX2 overexpression promoted cellular proliferation, decreased the percentage of cells in G0/G1 phase, and increased invasion of both U251 and U87 cells. Overexpression of TPX2 also significantly enhanced the phosphorylation of AKT, decreased the expression of p21, and increased the expression of cyclin D1 and matrix metallopeptidase (MMP)-9. In both U251 and U87 cells, knockdown of TPX2 resulted in phenotypes that are in direct contrast to those observed following TPX2 overexpression. Specifically, TPX2 knockdown inhibited cell proliferation, increased the percentage of cells in G0/G1 phase, inhibited invasion, decreased AKT phosphorylation, decreased the expression of MMP-9 and cyclin D1, and increased p21 expression. The AKT inhibitor IV in large part phenocopied the effect of TPX2 knockdown. The present data suggest that TPX2 promotes glioma cell proliferation and invasion via AKT signaling.