ER-ß accelerates the process of primary osteoporosis by promoting VEGFA-mediated apoptosis of osteoblasts.

Primary osteoporosis (POP) is a widespread and severe disorder of bone metabolism characterized by reduced bone mass and destruction of bone structure, frequently inducing fracture risk and imposing a heavy economic burden on public life. The development of POP partially revolves around the estrogen receptor ß (ER-ß), one of the major mediator receptors of estrogen that influences apoptosis in a range of cells. We performed KEGG and GO analysis by mining the transcriptomic dataset of POP samples showing significant enrichment of differentially expressed genes (DEGs) in multiple apoptosis-related pathways. The results of the Spearman correlation analysis and Protein-Protein Interaction (PPI) Networks screening of hub genes indicated that vascular endothelial growth factor A (VEGFA) may be a key target of ER-ß in controlling osteoblast apoptosis. Further, we carried out high-throughput sequencing of ESR2-silenced MC3T3-E1 cells and noticed a substantial suppression in VEGFA expression and all apoptosis-related pathways. In addition, we determined the cell cycle and apoptosis by constructing a VEGFA-silenced cell model utilizing flow cytometry (FCM), and the results showed that ER-ß could regulate the osteoblast cycle and thus promote osteoblast apoptosis by promoting VEGFA expression. And Western blot results showed that apoptosis was most likely realized through the regulation of downstream apoptosis markers c-JUN (c-Jun N-terminal kinase, JNK) and GADD45G (Growth Arrest and DNA Damage-Inducible Protein 45 gamma). The effects of ESR2 and VEGFA on the proliferation of osteoblasts were lastly assessed using the cell counting kit- 8 (CCK-8) assay. In conclusion, this study identifies that the roles of ER-ß in the regulation of osteoblast apoptosis are closely related to VEGFA and provides a new target for POP treatment.

The Association of nutritional status and physical activity on osteoporotic refractures among older adults.

BACKGROUND: This paper focuses on revealing the relationship between the Geriatric Nutritional Risk Index (GNRI) and Activity of Daily Living (ADL) with osteoporotic refracture. METHODS: Data from 1068 inpatients with osteoporotic fractures were analyzed. Binary logistic regression, Cox proportional hazard regression and Kaplan-Meier curves were performed for osteoporosis characteristics and its risk factors. Receiver operating characteristic (ROC) curve was developed to predict the cut-off value. RESULTS: The study showed that older age, lower ADL and lower GNRI were independent risk factors for osteoporotic fracture with OR of 1.039, 0.946, 0.892 and HR of 1.033, 0.967, 0.947 respectively. According to the results of ROC, the predictive accuracy of GNRI was high with an area under ROC (AUC) of 0.715, sensitivity of 76.6%, specificity of 53.5% and a threshold value of 99.65. CONCLUSION: Older age, lower ADL and lower GNRI were independent risk factors for osteoporotic refracture.

Smoking, alcohol consumption and risk of Dupuytren's disease: a Mendelian randomization study.

BACKGROUND: The correlation between smoking and alcohol consumption and the development of Dupuytren's disease (DD) has been acknowledged. However, the definitive causal relationship between these two factors and DD remains elusive. In order to establish a causal connection, we employed the two-sample Mendelian randomization method to evaluate the relationship between smoking and alcohol consumption and DD. METHODS: Based on publicly available genome-wide association studies (GWAS), two-sample univariate MR analyses were performed to assess the causal effects of drinks per week, cigarettes per day, smoking initiation, age of initiation, and smoking cessation on DD. We used inverse variance weighted (IVW) to generate the primary results for the MR analysis. Furthermore, we performed sensitivity MR analyses based on various methods to assess the robustness of estimations. Bidirectional MR analyses were used to study the interaction between smoking and alcohol consumption. Multivariate MR analyses were used to obtain independent causal effects of smoking or drinking on DD. RESULTS: Our two-sample MR, which was predominately based on IVW, revealed a causal relationship between drinks per week and DD (OR = 2.948, 95%CI: 1.746-4.975, P = 5.16E-05). In addition, there is no causal association between cigarettes per day, smoking initiation, age of initiation, smoking cessation and DD. Similar conclusions were reached by other MR methods. The results of the bidirectional MR analyses showed that the causal relationships between age of initiation and drinks per week were robust and significant. Multivariate MR results indicated that the causal effect of alcohol consumption on DD was independent of smoking. CONCLUSION: Our Mendelian Randomization study indicated that there is a causality between drinking alcohol and DD, but no such causality was found between smoking and DD. This is the first study to prove that drinking alcohol could cause DD. This could help people who are trying to prevent DD from happening in the first place.

Biological role of heparan sulfate in osteogenesis: A review.

Heparan sulfate (HS) is extensively expressed in cells, for example, cell membrane and extracellular matrix of most mammalian cells and tissues, playing a key role in the growth and development of life by maintaining homeostasis and implicating in the etiology and diseases. Recent studies have revealed that HS is involved in osteogenesis via coordinating multiple signaling pathways. The potential effect of HS on osteogenesis is a complicated and delicate biological process, which involves the participation of osteocytes, chondrocytes, osteoblasts, osteoclasts and a variety of cytokines. In this review, we summarized the structural and functional characteristics of HS and highlighted the molecular mechanism of HS in bone metabolism to provide novel research perspectives for the further medical research.

TRIM58 Interacts with Pyruvate Kinase M2 to Inhibit Tumorigenicity in Human Osteosarcoma Cells.

BACKGROUND: Tripartite motif containing 58 (TRIM58), an E3 ubiquitin ligase, is reported as a suppressor gene in certain human tumors. However, the biological function of TRIM58 in osteosarcoma (OS) is still less identified. METHODS: In the present study, TRIM58 induced silencing and overexpression in OS cells using RNA interference (RNAi) and lentiviral-mediated vector, respectively. Cell proliferation profiles were analyzed using cell counting kit-8 (CCK-8) assay. Cell apoptosis profiles were determined using a flow cytometer. qRT-PCR and western blot were used to determine gene expression. Coimmunoprecipitation (Co-IP) assay was used to examine protein interaction. RESULTS: Our results demonstrated TRIM58 was downregulated in human OS tissues. Overexpression of TRIM58 remarkably suppressed the growth of OS cells and decreased glucose transportation and lactate secretion. These results indicated that TRIM58 involved in the regulation of energy metabolism in OS cells. Importantly, TRIM58 interacted with pyruvate kinase M2 (PKM2) in OS cells. Moreover, TRIM58 might inhibit the activity of PKM2 through enhancing its polyubiquitination in OS cells. CONCLUSIONS: This analysis not only explored a deep understanding of the biological function of TRIM58 but also indicated its signaling pathway in OS cells.

Heparan sulfate loaded polycaprolactone-hydroxyapatite scaffolds with 3D printing for bone defect repair.

With the increasing applications of 3D printing technology in biomedical field, the composition or additives of the related materials has become critical for the next development. In the current study, we have prepared 3D printed polycaprolactone-hydroxyapatite (PCL-HA) porous scaffolds with loaded heparan sulfate (HS), in order to reveal the reparative effect of different concentrations of HS on the healing of bone defects. As a result, the scaffold itself showed sound compression resistance, air porosity and good biocompatibility. From both in vitro and in vivo experiments, the scaffold with low concentration of HS led to positive effects in promoting osteoblast maturation and accelerating bone defect repair. Moreover, scaffold with high concentration of HS showed notable inhibitive effort on the proliferation of osteoblasts, yet it still brought about positive effects in repairing bone defects in organisms. Thus, with good structural properties and biocompatibility, 3D printed PCL-HA-HS composite scaffold can facilitate and accelerate the repairing of biological bone defects, demonstrating as an effective biomaterial for bone defect repair.

Selective regulation of RANKL/RANK/OPG pathway by heparan sulfate through the binding with estrogen receptor ß in MC3T3-E1 cells.

Heparan sulfate (HS) is a linear anionic polysaccharide with repeating sulfated disaccharide units, which has been proven with various regulatory osteogenesis effects through multi-pathway signaling, but its impacts on receptor-activator of nuclear factor kappa beta ligand/receptor-activator of nuclear factor kappa beta/osteoprotegerin (RANKL/RANK/OPG) pathway is still poorly understood. In this study, the binding affinity between HS and estrogen receptor beta (ER-ß) was virtually analyzed using computer simulative docking method and experimentally studied by surface plasmon resonance (SPR). Thereafter, short interfering RNAs (siRNAs) were constructed to deliberately down-regulate the level of ER-ß in MC3T3-E1 cell line, and the transfected and non-transfected osteoblasts displaying good growth conditions were subsequently treated with HS. The results indicated that HS significantly reduced the expression level of RANKL without markedly affecting the expression of decoy receptor OPG during osteoblast differentiation, which can be partially owing to the interaction between HS and ER-ß. Meanwhile, the expression of RANKL in MC3T3-E1 cells was obviously increased after the transfection, demonstrating ER-ß as the key biomarker that regulates RANKL expression. The current work provided important supplementary information on the regulation mechanism of RANKL/RANK/OPG axis by HS.

Elevated expression of p53 gain-of-function mutation R175H in endometrial cancer cells can increase the invasive phenotypes by activation of the EGFR/PI3K/AKT pathway.

BACKGROUND: p53 is the most commonly mutated tumor suppressor gene in human cancers. In addition to the loss of tumor suppression function and exertion of dominant-negative effects over the remaining wild-type protein, several p53 mutants can gain novel oncogenic functions (gain-of-function, GOF) that actively regulate cancer development and progression. In human endometrial cancer, p53 mutation is more often associated with aggressive nonendometrioid cancer. However, it was unknown if p53 mutants contributed to endometrial cancer progression through the GOF properties. METHODS: To clarify the relationship between expression of p53 GOF mutation (p53-R175H) and invasive potential of human endometrial cancer KLE cells, we tested the consequences of up-regulation and down-regulation of p53-R175H in KLE cells by inducing p53-R175H expression vector or suppressing the p53 gene with short hairpin RNA. RESULTS: We found that forced over-expression of p53-R175H significantly promoted cell migration and invasion, and induced activation of the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/AKT pathway. Conversely, suppression of p53-R175H with short hairpin RNA significantly inhibited cell migration and invasion, and resulted in attenuation of EGFR/PI3K/AKT pathway. CONCLUSION: These findings show for the first time that elevated expression of p53-R175H mutant may exert gain-of-function activity to activate the EGFR/PI3K/AKT pathway and thus may contribute to the invasive phenotype in endometrial cancer.

Postoperative redislocation of the hip in a patient with congenital insensitivity to pain with anhidrosis: A case report and review of literature.

Pelvic osteotomy is commonly used to adjust acetabula dysplasia for congenital dislocation of the hip, whereas congenital insensitivity to pain with anhidrosis (CIPA) is a rare hereditary disease that often has the characteristics of joint development deformity and easy fracture. This article reports the case involving a CIPA patient who was surgically treated by Chiari pelvic osteotomy and proximal femoral rotation osteotomy for congenital dislocation of the left hip joint and was provided long-term follow-up for redislocation and bilateral femoral head absorption.

[Research progress in osteogenesis and osteogenic mechanism of heparan sulfate].

Objective: To discuss the role of heparan sulfate (HS) in bone formation and bone remodeling and summarize the research progress in the osteogenic mechanism of HS. Methods: The domestic and abroad related literature about HS acting on osteoblast cell line in vitro, HS and HS composite scaffold materials acting on the ani-mal bone defect models, and the effect of HS proteoglycans on bone development were summarized and analyzed. Results: Many growth factors involved in fracture healing especially heparin-binding growth factors, such as fibroblast growth factors, bone morphogenetic protein, and transforming growth factor ß, are connected noncovalently with long HS chains. HS proteoglycans protect these proteins from protease degradation and are directly involved in the regulation of growth factors signaling and bone cell function. HS can promote the differentiation of stem cells into osteoblasts and enhance the differentiation of osteoblasts. In bone matrix, HS plays a significant role in promoting the formation, maintaining the stability, and accelerating the mineralization. Conclusion: The osteogenesis of HS is pronounced. HS is likely to become the clinical treatment measures of fracture nonunion or delayed union, and is expected to provide more choices for bone tissue engineering with identification of its long-term safety.

Pravastatin attenuates left ventricular remodeling and diastolic dysfunction in angiotensin II-induced hypertensive mice.

BACKGROUND: A substantial proportion of patients with heart failure have a normal ejection fraction and diastolic dysfunction. However, there are few data available to guide the therapy of these patients. The effects of statins on cardiac remodeling are well documented in animal models and it is reported that statin therapy revealed a survival benefit in patients with diastolic heart failure (DHF). However, the exact mechanisms of statins possibly explaining the decreased cardiovascular morbidity and mortality in patients with DHF have not been elucidated. METHODS: We used 8-week-old male C57BL/6J mice, in which angiotensin II was subcutaneously infused for 4 weeks to mimic cardiac remodeling and fibrosis. They were treated with either normal saline or pravastatin in daily doses, which did not lower the serum cholesterol levels and blood pressure. RESULTS: Pravastatin improved diastolic dysfunction in angiotensin II-induced hypertensive mice, which was associated with the amelioration of left ventricular hypertrophy and remodeling. However, statin treatment showed no effect on the increased systolic blood pressure or cholesterol levels by angiotensin II infusion. The cardioprotective effects of pravastatin were closely associated with the downregulation of collagen I, transforming growth factor-beta, matrix metalloproteinases-2 and -3, atrial natriuretic factor, interleukin-6, tumor necrosis factor-alpha, ROCK1 gene expression, and the upregulation of endothelial nitric oxide synthase gene expression. CONCLUSIONS: The beneficial effects of pravastatin on DHF and structural remodeling are through cholesterol- independent mechanism of statins or "pleiotropic" effects of statins involving improving or restoring endothelial function and decreasing vascular inflammation. These findings suggest the potential involvement of ROCK1. Thus, treatment with pravastatin might be beneficial in patients with DHF.

Silencing of IQGAP1 by shRNA inhibits the invasion of ovarian carcinoma HO-8910PM cells in vitro.

BACKGROUND: IQGAP1 is a scaffolding protein and overexpressed in many human tumors, including ovarian cancer. However, the contribution of IQGAP1 to invasive properties of ovarian cancer cells remains unknown. Here, we investigated the effect of IQGAP1-specific short hairpin RNA (shRNA) expressing plasmids on metastatic potential of ovarian cancer HO-8910PM cells. METHODS: We used RT-PCR and Western blot analysis to characterize expression of IQGAP1 in three human ovarian cancer-derived cell lines SK-OV-3, HO-8910 and HO-8910PM. We then determined whether expression of endogenous IQGAP1 correlated with invasive and migratory ability by using an in vitro Matrigel assay and cell migration assay. We further knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter in HO-8910PM cells and examined the proliferation activity, invasive and migration potential of IQGAP1 shRNA transfectants using MTT assay, in vitro Matrigel-coated invasion assay and migration assay. RESULTS: IQGAP1 expression level seemed to be closely associated with the enhanced invasion and migration in ovarian cancer cell lines. Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs. RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration. CONCLUSION: Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.