Association between magnesium, copper, and potassium intakes with risk of rheumatoid arthritis: a cross-sectional study from National Health and Nutrition Examination Survey (NHANES).

BACKGROUND: The relationship between Mg (magnesium), Cu (copper), and K (potassium) intakes and the risk of rheumatoid arthritis (RA) remains limited. The aim of present study was to examine the associations between Mg, Cu and K intakes with RA. METHODS: Using data from the National Health and Nutrition Examination Survey (NHANES) 2003-2018, we examined the association between Mg, Cu and K intakes and the risk of RA among US adults. After adjustment for age, sex, race, BMI, educational level, smoking history, alcohol consumption, family Poverty Income Ratio (PIR), diabetes and total daily energy intake, logistic regression models and smooth curve fitting were applied to examine the associations of Mg, Cu and K intakes with RA. RESULTS: A total of 18,338 participants were included (1,008 participants with RA). The multivariate adjusted ORs (95% CI) of RA were [0.66 (0.51, 0.84)], [0.76 (0.60, 0.97)], and [0.75 (0.58, 0.97)] in the highest versus lowest quartile of magnesium intakes, respectively. A nonlinear association between Cu intakes and RA was found. When Cu intake (ln) was between 0.6-2.2 mg, the risk of RA reduced by 26% for every 1 mg increase of intake in Cu [0.74 (0.58, 0.96)]. CONCLUSIONS: Higher Mg, Cu and K intakes may be inversely associated with the risk of RA among US adults, and an inverse L-shaped association between dietary Cu and RA was found.

A novel molecular classification method for osteosarcoma based on tumor cell differentiation trajectories.

Subclassification of tumors based on molecular features may facilitate therapeutic choice and increase the response rate of cancer patients. However, the highly complex cell origin involved in osteosarcoma (OS) limits the utility of traditional bulk RNA sequencing for OS subclassification. Single-cell RNA sequencing (scRNA-seq) holds great promise for identifying cell heterogeneity. However, this technique has rarely been used in the study of tumor subclassification. By analyzing scRNA-seq data for six conventional OS and nine cancellous bone (CB) samples, we identified 29 clusters in OS and CB samples and discovered three differentiation trajectories from the cancer stem cell (CSC)-like subset, which allowed us to classify OS samples into three groups. The classification model was further examined using the TARGET dataset. Each subgroup of OS had different prognoses and possible drug sensitivities, and OS cells in the three differentiation branches showed distinct interactions with other clusters in the OS microenvironment. In addition, we verified the classification model through IHC staining in 138 OS samples, revealing a worse prognosis for Group B patients. Furthermore, we describe the novel transcriptional program of CSCs and highlight the activation of EZH2 in CSCs of OS. These findings provide a novel subclassification method based on scRNA-seq and shed new light on the molecular features of CSCs in OS and may serve as valuable references for precision treatment for and therapeutic development in OS.

MiR-30b-5p attenuates the inflammatory response and facilitates the functional recovery of spinal cord injury by targeting the NEFL/mTOR pathway.

BACKGROUND: Neurofilament light chain (NEFL) has been identified as a biomarker for spinal cord injury (SCI), but its effect and underlying mechanism in SCI remain unclear. METHODS: SCI rat models were established for in vivo studies. Lipopolysaccharide (LPS)-induced cell models were used for in vitro studies. The protein and mRNA expression levels of genes were evaluated by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The pathological changes in rats after SCI were subjected to histological examinations. The interaction of NEFL and upstream miRNAs was explored using dual-luciferase reporter gene assays. RESULTS: NEFL was highly expressed in SCI rat spinal cord tissues and LPS-stimulated PC12 cells. NEFL silencing showed an inhibitory effect on the morphological changes of SCI rats and the secretion of inflammatory factors and facilitated functional recovery of SCI rats. MiR-30b-5p was demonstrated to target NEFL and negatively regulate NEFL mRNA and protein levels. Downregulation of miR-30b-5p in SCI cell and rat models was demonstrated. MiR-30b-5p alleviated the inflammatory response in SCI rat models and LPS-stimulated PC12 cells and promoted functional recovery in rats by targeting NEFL. NEFL activated mTOR signaling. MiR-30b-5p inactivated mTOR signaling by negatively regulating NEFL. CONCLUSION: MiR-30b-5p alleviated the inflammatory response and facilitated the functional recovery of SCI rats by targeting NEFL to inactivate the mTOR pathway.

Association between Serum Cotinine Levels and Bone Mineral Density: An Analysis of the National Health and Nutrition Examination Survey (NHANES).

Purpose: To investigate the relationship between serum cotinine and lumbar bone mineral density (BMD) among 7905 participants aged 30 years and over. Method: A total of 3945 men and 3960 women from the National Health and Nutrition Examination Survey 2011-2018 were included in this cross-sectional analysis. Independent variable was serum cotinine, which is a biomarker of cigarette exposure. The outcome variable was lumbar BMD. We investigated the associations of serum cotinine levels and lumbar BMD using multivariable linear regression models. Results: Serum cotinine concentration was negatively associated with lumbar BMD after adjustment of relevant covariables (ß = -0.039, 95% CI: -0.078 to -0.014, P = 0.005). However, in the subgroup analysis stratified by gender, this negative association remained only in women (ß = -0.072, 95% CI: -0.132 to -0.012, P = 0.019). Conclusion: Our study suggested that elevated serum cotinine level correlated with decreased lumbar BMD, especially in women. This finding indicated that reducing cigarette exposure and maintaining serum cotinine at a low level may be beneficial to bone health for adults.

REGγ drives Lgr5+ stem cells to potentiate radiation induced intestinal regeneration.

Leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), a marker of intestinal stem cells (ISCs), is considered to play key roles in tissue homoeostasis and regeneration after acute radiation injury. However, the activation of Lgr5 by integrated signaling pathways upon radiation remains poorly understood. Here, we show that irradiation of mice with whole-body depletion or conditional ablation of REGγ in Lgr5+ stem cell impairs proliferation of intestinal crypts, delaying regeneration of intestine epithelial cells. Mechanistically, REGγ enhances transcriptional activation of Lgr5 via the potentiation of both Wnt and Hippo signal pathways. TEAD4 alone or cooperates with TCF4, a transcription factor mediating Wnt signaling, to enhance the expression of Lgr5. Silencing TEAD4 drastically attenuated ß-catenin/TCF4 dependent expression of Lgr5. Together, our study reveals how REGγ controls Lgr5 expression and expansion of Lgr5+ stem cells in the regeneration of intestinal epithelial cells. Thus, REGγ proteasome appears to be a potential therapeutic target for radiation-induced gastrointestinal disorders.

Systematic identification of the key candidate genes in breast cancer stroma.

BACKGROUND: Tumor microenvironment, in particular the stroma, plays an important role in breast cancer cell invasion and metastasis. Investigation of the molecular characteristics of breast cancer stroma may reveal targets for future study. METHODS: The transcriptome profiles of breast cancer stroma and normal breast stroma were compared to identify differentially expressed genes (DEGs). The method was analysis of GSE26910 and GSE10797 datasets. Common DEGs were identified and then analyses of enriched pathways and hub genes were performed. RESULTS: A total of 146 DEGs were common to GSE26910 and GSE10797. The enriched pathways were associated with "extracellular matrix (ECM) organization", "ECM-receptor interaction" and "focal adhesion". Network analysis identified six key genes, including JUN, FOS, ATF3, STAT1, COL1A1 and FN1. Notably, COL1A1 and FN1 were identified for the first time as cancer stromal key genes associated with breast cancer invasion and metastasis. Oncome analysis showed that the high expression levels of COL1A1 and FN1 correlated to an advanced stage of breast cancer and poor clinical outcomes. CONCLUSIONS: We found that several conserved tumor stromal genes might regulate breast cancer invasion through ECM remodeling. The clinical outcome analyses of COL1A1 and FN1 suggest these two genes are promising targets for future studies.

AMPH-1 is critical for breast cancer progression.

Amphiphysin 1 (AMPH-1) is a nerve terminals-enriched protein involved in endocytosis, and we observe that its expression is increased in breast cancer tumor in compared with normal breast. However, its function in breast cancer is unknown. Here we aim to explore the role of AMPH-1 in breast cancer cells. Knockdown of AMPH-1 in breast cancer cells promotes cell proliferation, cell cycle progression and cell migration, and attenuates cell apoptosis. Of note, knockdown of AMPH-1 promotes breast cancer progression in xenograft mouse model. These oncogenic phenotypes may be partially due to the activated EMT and ERK pathways after inhibition of AMPH-1. Oncomine analyses of multiple breast cancer patient datasets show that reduced AMPH-1 mRNA level is significantly associated with breast cancer patients having metastatic events, advanced stage, poor clinical outcomes, and Paclitaxel+FEC treatment resistance. In summary, our results identified the anti-oncogenic function of AMPH-1 in breast cancer in vitro and in vivo. Activation of AMPH-1 may be a promising approach to treat breast cancer patients.

miR-215 suppresses proliferation and migration of non-small cell lung cancer cells.

The expression of microRNA-215 (miR-215) in non-small cell lung cancer (NSCLC) tissues and the effects of miR-215 on the proliferation and migration of NSCLC cells were investigated. qRT-PCR was used to detect the expression of miR-215 in NSCLC tissues and paired normal tumor-adjacent lung tissues; MTT assay, transwell assay and soft-agar assay were used in vitro to evaluate the role of miR-215 on proliferation, migration and cell clonality on NSCLC cells, after transfecting miR-215 mimics to NSCLC cell line A549 or miR-215 to H1299. miR-215 was significantly decreased in NSCLC tissues compared to the paired normal tissues; Overexpression of miR-215 in A549 cells resulted in reduction of the cell proliferation, migration and cell clonality, while downregulation of miR-215 in H1299 cells could promote cell proliferation, migration and clonality. In conclusion, miR-215 was downregulated in NSCLC tissues and may play a key role in the development of NSCLC.

Preferentially Expressed Antigen of Melanoma Prevents Lung Cancer Metastasis.

Lung cancer is the most common cause of cancer death worldwide. The poor survival rate is largely due to the extensive local invasion and metastasis. However, the mechanisms underlying the invasion and metastasis of lung cancer cells remain largely elusive. In this study, we examined the role of preferentially expressed antigen of melanoma (PRAME) in lung cancer metastasis. Our results show that PRAME is downregulated in lung adenocarcinoma and lung bone metastasis compared with normal human lung. Knockdown of PRAME decreases the expression of E-Cadherin and promotes the proliferation, invasion, and metastasis of lung cancer cells by regulating multiple critical genes, most of which are related to cell migration, including MMP1, CCL2, CTGF, and PLAU. Clinical data analysis reveals that the expression of MMP1 correlates with the clinical features and outcome of lung adenocarcinoma. Taken together, our data demonstrate that PRAME plays a role in preventing the invasion and metastasis of lung adenocarcinoma and novel diagnostic or therapeutic strategies can be developed by targeting PRAME.

Identification of Preferentially Expressed Antigen of Melanoma as a Potential Tumor Suppressor in Lung Adenocarcinoma.

BACKGROUND Preferentially expressed antigen of melanoma (PRAME) is known as a tumor-associated antigen that is altered in a variety of malignancies, including lung cancer. However, the role of PRAME in lung cancer remains unclear. MATERIAL AND METHODS We analyzed the expression of PRAME in human lung adenocarcinomas and studied the function of PRAME using small interfering RNA (siRNA)-induced gene knockdown in lung cancer cell lines PC9 and A549. RESULTS We found that PRAME expression is down-regulated in lung adenocarcinomas. Knockdown of PRAME promoted proliferation and suppressed apoptosis of PC9 and A549 cells. CONCLUSIONS In line with its roles in controlling cell growth, RPAME regulates multiple critical cell-growth related genes, including IGF1R oncogene. IGF1R up-regulation contributes to increase of cell growth upon the knockdown of PRAME. Taken together, our results suggest that PRAME has inhibitory roles in lung cancer.

Clinical Features and Prognostic Factors of Pediatric Spine Tumors: A Single-Center Experience With 190 Cases.

STUDY DESIGN: A retrospective study was performed. OBJECTIVE: The aim of the study was to illustrate the characteristics of pediatric spine tumors (PSTs) and obtain better insight into therapeutic strategies of such kinds of tumors. SUMMARY OF BACKGROUND DATA: PSTs, a heterogeneous group of tumors occurred among pediatric group, are relatively uncommon and rarely reported in the literature. METHODS: A retrospective study was performed to analyze clinical features and prognostic factors of patients with PSTs who were treated in our center between 2000 and 2013. Local relapse-free survival (LRFS) and overall survival (OS) rate were estimated using the Kaplan-Meier method to identify potential prognostic factors. Factors with P values of 0.1 or lower were subjected to multivariate analysis by Cox regression analysis. P values of 0.05 or lower were considered statistically significant. RESULTS: A total of 190 patients with PSTs were included in the study. The mean follow-up period was 53.5 months. Of the 190 patients, 127 cases (66.8%) were diagnosed as benign lesions or tumor-like lesions, with 19 cases as aggressive lesions and 44 cases as malignant lesions. Recurrence was detected in 30 patients after initial surgery in our center, whereas death occurred in 24 cases. Implant failure and spinal deformity occurred in 3 and 5 patients, respectively. The statistical analysis suggested that multiple segments involvement (n ≥ 3), aggressive and malignant lesion were independent prognostic factors for LRFS, whereas malignant lesion was the only unfavorable factors for OS. In the subgroup analysis, multiple segments involvement (n ≥ 3) and Enneking stages III were independent factors for LRFS of patients with benign or aggressive lesions. CONCLUSION: Malignant lesion was significantly associated with decreased OS and LRFS of PSTs, whereas multiple segments involvement (n ≥ 3) and aggressive lesions were significantly associated with LRFS. LEVEL OF EVIDENCE: 4.

TGF-ß1-induced miR-202 mediates drug resistance by inhibiting apoptosis in human osteosarcoma.

PURPOSE: A number of studies have demonstrated that microRNAs play a critical role in osteosarcoma progression, therapy and drug resistance. MicroRNA-202 has proven to be dysregulated in many human cancer studies. This study aimed to explore miR-202 contributions to drug resistance in osteosarcoma. METHODS: miR-202 expression was measured by real-time PCR in patient tissues, cell lines or cells treated with TGF-ß1, using siRNA to knock down the expression of Smad2, Smad3 and Smad4. miR-202 mimics, inhibitor and scramble siRNA were transfected into osteosarcoma cells to observe effects on cell apoptosis and drug resistance. Moreover, relationships of miR-202 level with PDCD4 were investigated by luciferase reporter assay, real-time PCR and Western blotting. RESULTS: We found that miR-202 is overexpressed in osteosarcoma tissues when compared with normal human osteoblasts and TGF-ß1 can induce the expression of miR-202. Transfection of miR-202 mimics into osteosarcoma cell lines significantly promotes chemotherapy resistance by targeting PDCD4, a tumor suppressor which is involved in apoptosis. In contrast, transfection of miR-202 inhibitor enhances the drug sensitivity and apoptosis. CONCLUSIONS: This study provides new insights into miRNA-202 in osteosarcoma as a potential molecular target for chemotherapy.

5-Fluoruracil blocked giant cell tumor progression by suppressing osteoclastogenesis through NF-kappaB signals and blocking angiogenesis.

Giant cell tumor of bone (GCTB) is a bone destroying tumor comprised of spindle-like stromal cells and monocytes of myeloid lineage that are differentiated into osteoclast-like multinucleated giant cells. Nuclear factor-Kappa B (NF-κB) has been identified to be essential for GCT progression. Herein, we found that 5-Fluorouracil (5-FU), a widely used chemotherapeutics, is a promising anticancer agent for GCT both targeting spindle-like stromal cells and osteoclast giant cells through NF-κB pathway. In this study, in vitro 5-FU not only directly blocked both stromal cell- and RANKL-induced osteoclastogenesis through NF-κB pathway, but also indirectly inhibited osteoclast formation and angiogenesis by suppressing the expression of osteoclast-activating factors including IL-1ß, MCP-1 and tumor angiogenesis factor VEGF in stromal cells. In vivo, we found that 5-FU blocked GCT progression through NF-κB pathway by utilizing our chick embryo chorioallantoic membrane (CAM) model. Taken together, our results suggest that 5-FU can inhibit GCT development by suppressing osteoclast formation through NF-κB pathway and blocking angiogenesis, and may serve as a novel agent in the treatment of GCT.

Primary chordomas of the cervical spine: a consecutive series of 14 surgically managed cases.

OBJECT: Cervical chordomas are rare lesions and usually bring about challenges in treatment planning because of their wide extension and complicated adjacent anatomy. There are few large published series at present focusing on cervical chordomas. The authors studied a consecutive series of 14 patients with primary cervical chordomas who underwent surgical treatment and were observed between 1989 and 2008. By reviewing the clinical patterns and follow-up data, they sought to investigate the clinical characters, tailor the appropriate surgical techniques, and establish prognosis factors for cervical chordomas. METHODS: Hospitalization and follow-up data in the 14 patients were collected. All patients underwent piecemeal tumor excision and reconstruction for stability; total spondylectomy was achieved in 5 cases. Postoperative radiotherapy was administered in all patients. Kaplan-Meier plots were used to represent tumor recurrence and patient survival, and log-rank testing was used to determine the risk factors of local recurrence. RESULTS: Follow-up ranged from 8 to 120 months (mean 58.6 months). Symptom and neural status in most patients improved after surgery. The 1- and 5-year disease-free survival rates were 78.6% and 50%, respectively, and the 1- and 5-year survival rates were 92.9% and 85.7%, respectively. Log-rank tests revealed that the following variables were significantly associated with a high rate of tumor recurrence: age less than 40 years or greater than 70 years (p = 0.006) and an upper cervical tumor location (p = 0.019). CONCLUSIONS: Chordomas in the cervical spine are usually neoplasms that exhibit insidious growth and a wide extension by the time of diagnosis. Radical intralesional debulking surgery and postoperative radiotherapy have been effective treatment. A limited application of en bloc tumor resection and the highly likely intraoperative intralesional tumor seeding may partially explain the high local recurrence rate, whereas the chance of distant metastases, fortunately, is very low. Most recurrence were documented within 3 years. Some specific surgical techniques should be emphasized to minimize tumor seeding. Patients with upper cervical chordomas, younger adults, and elderly adults have worse prognosis. For patients with chordoma extending to both the anterior and posterior spinal columns, total spondylectomy combined with piecemeal excision is recommended for a better prognosis.

[The effect of HA mixed with adenovirus mediated rhBMP-2 transferred BMSCs of goats on distraction osteogenesis].

OBJECTIVE: To evaluate the effectiveness of HA mixed with adenovirus mediated rhBMP-2 gene (Adv-rhBMP-2) transferred BMSCs of goats on distraction osteogenesis. METHODS: Nineteen adult goats were used for the experiment, no matter they were male or female, and the weight of the goats were 15-20 kg. The 10 mL marrow was obtained from the iliac crest of each goat. The BMSCs was expanded and passaged conventionally. The 3th BMSCs was infected by Adv-rhBMP-2 at 200 multiplicity of infection (MOI). The 1 x 10(8) infected BMSCs were digested by 0.25% trypsin and collected, then mixed with HA. The right tibia lengthening models were developed, and mixture with BMSCs was injected in the osteotomy position. The goats were divided randomly into 4 groups according to the material injected in operation, group A: Adv-rhBMP-2/BMSCs/HA (n = 6); group B: Adv-rhBMP-2/BMSCs (n = 5); group C: Adv-beta-gal/BMSCs/HA (n = 4); group D: sham without any injections (n = 4). After a seven-day latency period following ostectomy, distraction was carried out at a rate of 1 mm/day for 4 weeks. Roentgenography was practiced in 5, 8 and 12 weeks. After 12 weeks, the goats were sacrificed and dual-energy X-ray absorptiometry (DXA), biomechanical test and histology results were studied. RESULTS: After five and eight weeks surgery, X-ray test showed the distraction callus was more in group A and B than group C and D, and the radiographic score was significantly higher in group A and B than in the other two groups (P < 0.05); after 12 weeks surgery, the continued callus was formed in the distraction defects in all groups. DXA showed the mean bone mineral content of distraction callus in group A, B, C, D was (4.175 +/- 1.921), (2.600 +/- 0.638), (2.425 +/- 0.826) and (1.175 +/- 0.574) g, and the mean bone mineral density was (0.612 +/- 0.196), (0.630 +/- 0.159), (0.450 +/- 0.166) and (0.266 +/- 0.113) g/cm2. The group A and B was significantly higher than group C and D (P < 0.05). Biomechanical test showed the maximum loading of group A, B, C, D was (490.20 +/- 155.08), (350.59 +/- 80.48), (221.95 +/- 68.79) and (150.65 +/- 92.29) N, and elastic modulus was (178.24 +/- 105.80), (105.88 +/- 27.09), (81.18 +/- 48.67) and (50.35 +/- 47.64) MPa. The group A was significantly higher than in group C and D (P < 0.05). Histology observation revealed abundant bone formation in the distraction defects in group A, and the bone trabecula was arranged longitudinal and netlike. Histomorphology analysis revealed the bone volume in group A, B, C, D was 72.35% +/- 5.68%, 67.58% +/- 7.42%, 49.63% +/- 4.87% and 38.87% +/- 2.35%, and the bone formation was significantly greater in group A compared with group D (P < 0.05). CONCLUSION: HA mixed with rhBMP-2 modified BMSCs can accelerate distraction osteogenesis in goats.