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Find video protocols related to scientific articles indexed in Pubmed.
Effect of the disruption of three cytoskeleton components on chondrocyte metabolism in rabbit knee cartilage.
Chin. Med. J.
PUBLISHED: 11-11-2014
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Chondrocytes' phenotype and biosynthesis of matrix are dependent on having an intact cytoskeletal structure. Microfilaments, microtubules, and intermediate filaments are three important components of the cytoskeletal structure of chondrocytes. The aims of this study were to determine and compare the effects of the disruption of these three cytoskeletal elements on the apoptosis and matrix synthesis by rabbit knee chondrocytes in vitro.
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Protective effects of resveratrol on postmenopausal osteoporosis: regulation of SIRT1-NF-?B signaling pathway.
Acta Biochim. Biophys. Sin. (Shanghai)
PUBLISHED: 11-06-2014
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Postmenopausal osteoporosis severely jeopardizes human health. Seeking for therapeutic drugs without side effects is of great necessity. Our study was designed to investigate whether resveratrol, an agonist of SIRT1, could have favorable effect on osteoporosis and to explore the underlying mechanisms. Rat osteoporosis model (ovariectomy group, OVX) was established by bilateral ovariectomy. Three different doses of resveratrol were used: 5 mg/kg/d (low-dosed, RES(LD)), 25 mg/kg/d (medium-dosed, RES(MD)), and 45 mg/kg/d (high-dosed, RES(HD)). Results showed that RES(LD) did not show any significant effect on OVX alterations, while RES(MD) and RES(HD) significantly elevated the decreased bone mineral density induced by osteoporosis (RES(MD) 0.205±0.023, RES(HD) 0.214 ± 0.053 vs. OVX 0.165 ± 0.050, g/cm(2) respectively; P < 0.05). Serum markers alkaline phosphatase (ALP) and osteocalcin were moderately restored by resveratrol. Moreover, resveratrol improved bone structure in OVX rats, demonstrated by hematoxylin-eosin staining and micro-computed tomographic results. In vitro results revealed that resveratrol promoted osteoblast differentiation of bone marrow mesenchymal stromal cells, evidenced by the increase of ALP generation and mRNA expression of collagen 1 (P < 0.05; RES(MD), RES(HD) vs. control group). SIRT1 gene silencing by siRNA transfection blocked these beneficial effects of resveratrol (P < 0.05; RES+ SIRT1(KD) vs. RES(HD)). Western blot results showed that resveratrol activated SIRT1 and subsequently suppressed the activity of NF-?B with decreased expression level of p-I?B? and NF-?B p65 (P < 0.05). Our findings verified the effects of specific dosed resveratrol on postmenopausal osteoporosis through osteoblast differentiation via SIRT1-NF-?B signaling pathway. This study suggested the therapeutic potential of resveratrol against osteoporosis and stressed the importance of effective doses.
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A Novel Total Cervical Prosthesis for Single-level Cervical Subtotal Corpectomy: Radiological and Histomorphometric Analysis in a Caprine Model.
J Spinal Disord Tech
PUBLISHED: 10-30-2014
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A novel total cervical prosthesis for single level cervical subtotal corpectomy was assessed in a caprine animal model.
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Roles of microRNA in vascular diseases in cardiac and pulmonary systems.
Pharmazie
PUBLISHED: 10-03-2014
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Coronary artery disease (CAD) and pulmonary arterial hypertension (PAH) are two of the most dangerous vascular diseases. Their etiology and pathogenesis are not yet fully understood, thus it remains difficult to achive great advance in the diagnose, therapy and prognosis techniques. microRNAs (miRNAs), a class of highly conserved, small, noncoding RNAs, critically mediate the post-transcriptional gene modulation, which regulates an array of important physiopathological processes including those occurring in cardiac and pulmonary systems. Thereby manipulation of miRNA expression could potentially be applied therapeutically. In this review, we summarize the current knowledge on the roles of miRNAs in the development of vascular diseases, especially in CAD and PAH, providing a theoretical basis for potential uses of miRNA in diagnosis, prognosis, and therapy in these cardiovascular diseases.
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Cross-species transcriptomic approach reveals genes in hamster implantation sites.
Reproduction
PUBLISHED: 09-24-2014
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The mouse model has greatly contributed to understanding molecular mechanisms involved in the regulation of progesterone (P4) plus estrogen (E)-dependent blastocyst implantation process. However, little is known about contributory molecular mechanisms of the P4-only-dependent blastocyst implantation process that occurs in species such as hamsters, guineapigs, rabbits, pigs, rhesus monkeys, and perhaps humans. We used the hamster as a model of P4-only-dependent blastocyst implantation and carried out cross-species microarray (CSM) analyses to reveal differentially expressed genes at the blastocyst implantation site (BIS), in order to advance the understanding of molecular mechanisms of implantation. Upregulation of 112 genes and downregulation of 77 genes at the BIS were identified using a mouse microarray platform, while use of the human microarray revealed 62 up- and 38 down-regulated genes at the BIS. Excitingly, a sizable number of genes (30 up- and 11 down-regulated genes) were identified as a shared pool by both CSMs. Real-time RT-PCR and in situ hybridization validated the expression patterns of several up- and down-regulated genes identified by both CSMs at the hamster and mouse BIS to demonstrate the merit of CSM findings across species, in addition to revealing genes specific to hamsters. Functional annotation analysis found that genes involved in the spliceosome, proteasome, and ubiquination pathways are enriched at the hamster BIS, while genes associated with tight junction, SAPK/JNK signaling, and PPAR?/RXR? signalings are repressed at the BIS. Overall, this study provides a pool of genes and evidence of their participation in up- and down-regulated cellular functions/pathways at the hamster BIS.
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Proteomic and metabolomic responses of Pacific oyster Crassostrea gigas to elevated pCO2 exposure.
J Proteomics
PUBLISHED: 08-28-2014
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The gradually increased atmospheric CO2 partial pressure (pCO2) has thrown the carbonate chemistry off balance and resulted in decreased seawater pH in marine ecosystem, termed ocean acidification (OA). Anthropogenic OA is postulated to affect the physiology of many marine calcifying organisms. However, the susceptibility and metabolic pathways of change in most calcifying animals are still far from being well understood. In this work, the effects of exposure to elevated pCO2 were characterized in gills and hepatopancreas of Crassostrea gigas using integrated proteomic and metabolomic approaches. Metabolic responses indicated that high CO2 exposure mainly caused disturbances in energy metabolism and osmotic regulation marked by differentially altered ATP, glucose, glycogen, amino acids and organic osmolytes in oysters, and the depletions of ATP in gills and the accumulations of ATP, glucose and glycogen in hepatopancreas accounted for the difference in energy distribution between these two tissues. Proteomic responses suggested that OA could not only affect energy and primary metabolisms, stress responses and calcium homeostasis in both tissues, but also influence the nucleotide metabolism in gills and cytoskeleton structure in hepatopancreas. This study demonstrated that the combination of proteomics and metabolomics could provide an insightful view into the effects of OA on oyster C. gigas.
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MicroRNA Regulates Vascular Endothelial Growth Factor Expression in Chondrosarcoma Cells.
Clin. Orthop. Relat. Res.
PUBLISHED: 08-09-2014
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Systemic treatments to prevent or treat chondrosarcoma metastasis are lacking and targeted therapy has yet to be developed. Hypoxia develops in tumors as they grow and hypoxia-related alterations in gene expression underlie some of the traits of cancer. One critical trait is the ability to induce sustained angiogenesis, which is usually related to expression of vascular endothelial growth factor (VEGF). A potential hypoxia-related mechanism resulting in altered gene expression involves microRNA. Little is known about microRNA expression in chondrosarcoma and its potential role in regulation of VEGF expression.
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Influence of urate-lowering therapies on renal handling of uric acid.
Clin. Rheumatol.
PUBLISHED: 08-07-2014
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The purpose of this study is to investigate the effect of urate-lowering therapies (ULTs) on renal uric acid excretion in gout patients. This prospective observational study involved 106 primary gout patients and 51 healthy controls. Gout patients received ULT with either xanthine oxidase inhibitors or the uricosuric agent benzbromarone. Parameters such as 24-h urinary uric acid, creatinine clearance, uric acid clearance, glomerular filtration load of uric acid, fractional excretion of uric acid, excretion of uric acid per volume of glomerular filtration, and urinary uric acid to urinary creatinine ratio were used to evaluate the pre- and post-treatment renal capacity for uric acid clearance in gout patients and were compared with the values in the healthy controls. Compared to healthy controls, gout patients had higher glomerular filtration load of uric acid and lower uric acid clearance, creatinine clearance, and fractional uric acid excretion. After ULT, both the xanthine oxidase inhibitor group and benzbromarone group patients showed reduction in glomerular filtration load of uric acid. Creatinine clearance was significantly improved in the xanthine oxidase inhibitor group. Excretion function was remarkably enhanced in patients who reached the treatment target (serum uric acid <6 mg/dl). Changes in glomerular uric acid filtration load were significantly correlated with changes in serum urate levels. Gout patients have impaired renal uric acid excretion. ULTs reduce renal urate load and enhance the renal capacity of uric acid clearance. Xanthine oxidase inhibitors showed superiority over benzbromarone in improving renal function.
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Scaled diffraction calculation between tilted planes using nonuniform fast Fourier transform.
Opt Express
PUBLISHED: 08-05-2014
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A method of diffraction calculation between tilted planes with variable sampling rates is proposed. The proposed method is based on the Fourier spectrum rotation from a tilted plane to a parallel plane. The nonuniform fast Fourier transformation (NUFFT) is used to calculate the nonuniform sampled Fourier spectrum on the tilted plane with variable sampling rates, which overcomes the sampling restriction of FFT in the conventional method. Both of the computer simulation and the optical experiment shows the feasibility of our method in calculating the hologram of polygon-based object with scalable size, which can be considered as an important application in the holographic three-dimensional display.
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The protective effect of curcumin on hepatotoxicity and ultrastructural damage induced by cisplatin.
Ultrastruct Pathol
PUBLISHED: 07-31-2014
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We investigate the protective effect of curcumin (CU) on the hepatic ultrastructural damage induced by cisplatin in mice. 18 adult Kunming mice were randomly divided into normal saline (NS) group, cisplatin treatment group (CP) and CU?+?CP group (n?=?6 for each group). Mice in control group and CP group were administered with NS (20?mL/kg/day) and CU?+?CP group were i.p injected with CU (200?mg/kg/day) for 10 days. Then cisplatin (50?mg/kg/day) was injected in mice of CP group and CU?+?CP group, while those in control group were given the same volume of NS. Five days after injection all mice were killed and liver dissected. The hepatic morphological structures were observed under light microscope and transmission electron microscope. The results indicated that CU alleviated the hepatic histopathological damages induced by cisplatin, which included declined body weight, vacuolated cytoplasm and blurred liver trabecular structure. Moreover, no hepatic ultrastructural damages were observed in the CU protective group with condensed and marginated nuclear chromatin, bile canaliculi outstreched and bile deposited.
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Ritonavir-induced hepatotoxicity and ultrastructural changes of hepatocytes.
Ultrastruct Pathol
PUBLISHED: 07-31-2014
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To investigate the effect of ritonavir on hepatocyte proliferation, we detected the change of cleaved caspase-3 expression level in the hepatocytes. Furthermore, the morphological and ultrastructural changes of hepatocytes derived from RTV-treated mice have been observed. The results showed that ritonavir can evidently inhibit hepatocyte proliferation and increase cleaved caspase-3 expression level. Under the electron microscope, chromatin margination, mitochondrial cristae disappearance, karyopyknosis and cytoplasmic vacuolization can be observed in the hepatocytes of mice treated with ritonavir. In conclusion, the mechanism of ritonavir's hepatotoxicity is that it induces apoptosis of hepatocytes via the caspase-cascade system.
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A one-pot synthetic strategy for construction of the dibenzodiazepine skeleton via a transition metal-free process.
Org. Biomol. Chem.
PUBLISHED: 07-25-2014
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A one-pot transition metal-free methodology for constructing pharmacologically active dibenzodiazepine derivatives was developed. Fluoro-, bromo- and nitro-substituted aryl aldehydes were applied to this reaction efficiently.
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Osseointegration of chitosan coated porous titanium alloy implant by reactive oxygen species-mediated activation of the PI3K/AKT pathway under diabetic conditions.
Biomaterials
PUBLISHED: 07-03-2014
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Chitosan coated porous titanium alloy implant (CTI) is demonstrated a promising approach to improve osseointegration capacity of pure porous titanium alloy implant (TI). Since chitosan has been demonstrated to exhibit antioxidant activity, we propose CTI may ameliorate the ROS overproduction, thus reverse the poor osseointegration under diabetic conditions, and investigate the underlying mechanisms. Primary rat osteoblasts incubated on the TI and the CTI were subjected to normal serum (NS), diabetic serum (DS), DS + NAC (a potent ROS inhibitor) and DS + LY294002 (a PI3K/AKT-specific inhibitor). In vivo study was performed on diabetic sheep implanted with TI or CTI into the bone defects on crista iliaca. Results showed that diabetes-induced ROS overproduction led to osteoblast dysfunction and apoptosis, concomitant with the inhibition of AKT in osteoblasts on the TI substrate. While CTI stimulated AKT phosphorylation through ROS attenuation, thus reversed osteoblast dysfunction evidenced by improved osteoblast adhesion, increased proliferation and ALP activity, and decreased cytotoxicity and apoptotic rate, which exerted same effect to NAC treatment on the TI. These effects were further confirmed by the improved osseointegration within the CTI in vivo evidenced by Micro-CT and histological examinations. In addition, the aforementioned promotive effects afforded by CTI were abolished by blocking PI3K/AKT pathway with addition of LY294002. These results demonstrate that the chitosan coating markedly ameliorates diabetes-induced impaired bio-performance of TI via ROS-mediated reactivation of PI3K/AKT pathway, which elicits a new surface functionalization strategy for better clinical performance of titanium implant in diabetic patients.
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FDG PET-CT combined with TBNA for the diagnosis of atypical relapsing polychondritis: report of 2 cases and a literature review.
J Thorac Dis
PUBLISHED: 06-27-2014
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To explore the value of (18)F-fluorodeoxyglucose positron emission tomography-computed tomography (FDG PET-CT) combined with transbronchial needle aspiration (TBNA) in diagnosing atypical relapsing polychondritis (RP).
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Representing images using curvilinear feature driven subdivision surfaces.
IEEE Trans Image Process
PUBLISHED: 06-25-2014
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This paper presents a subdivision-based vector graphics for image representation and creation. The graphics representation is a subdivision surface defined by a triangular mesh augmented with color attribute at vertices and feature attribute at edges. Special cubic B-splines are proposed to describe curvilinear features of an image. New subdivision rules are then designed accordingly, which are applied to the mesh and the color attribute to define the spatial distribution and piecewise-smoothly varying colors of the image. A sharpness factor is introduced to control the color transition across the curvilinear edges. In addition, an automatic algorithm is developed to convert a raster image into such a vector graphics representation. The algorithm first detects the curvilinear features of the image, then constructs a triangulation based on the curvilinear edges and feature attributes, and finally iteratively optimizes the vertex color attributes and updates the triangulation. Compared with existing vector-based image representations, the proposed representation and algorithm have the following advantages in addition to the common merits (such as editability and scalability): 1) they allow flexible mesh topology and handle images or objects with complicated boundaries or features effectively; 2) they are able to faithfully reconstruct curvilinear features, especially in modeling subtle shading effects around feature curves; and 3) they offer a simple way for the user to create images in a freehand style. The effectiveness of the proposed method has been demonstrated in experiments.
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Specific inhibition of HDAC4 in cardiac progenitor cells enhances myocardial repairs.
Am. J. Physiol., Cell Physiol.
PUBLISHED: 06-18-2014
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We have recently shown that in vivo inhibition of histone deacetylase (HDAC) stimulates endogenous myocardial regeneration in infarcted hearts (Zhang L et al. J Pharmacol Exp Ther 341: 285-293, 2012). Furthermore, our observation demonstrates that HDAC inhibition promotes cardiogenesis, which is associated with HDAC4 reduction. However, it remains unknown as to whether specific inhibition of HDAC4 modulates cardiac stem cells (CSCs) to facilitate myocardial repair and to preserve cardiac performance. c-kit(+) CSCs were isolated from adult mouse hearts and were transfected with HDAC4 siRNA to knockdown HDAC4 of c-kit(+) CSCs. The transfection of HDAC4 siRNA caused a marked reduction of HDAC4 mRNA and proteins in c-kit(+) CSCs. Mouse myocardial infarction (MI) was created to assess the effect of HDAC4 inhibition in c-kit(+) CSCs on myocardial regeneration in vivo when cells were introduced into MI hearts. Transplantation of HDAC4 siRNA-treated c-kit(+) CSCs into MI hearts improved ventricular function, attenuated ventricular remodeling, and promoted CSC-derived regeneration and neovascularization. Furthermore, Ki67 and BrdU positively proliferative myocytes increased in MI hearts receiving HDAC4 siRNA-treated c-kit(+) CSCs compared with MI hearts engrafted with control siRNA-treated c-kit(+) CSCs. In addition, compared with MI hearts engrafted with control adenoviral GFP-infected c-kit(+) CSCs, MI hearts receiving adenoviral HDAC4-infected c-kit(+) CSCs exhibited attenuated cardiac functional recovery, CSC-derived regeneration, and neovascularization, which was accompanied with adverse ventricular remodeling and decrease in Ki67 and BrdU positively proliferative myocytes. HDAC4 inhibition facilitated c-kit(+) CSCs into the differentiation into cardiac lineage commitments in vitro, while HDAC4 overexpression attenuated c-kit(+) CSC-derived cardiogenesis. Our results indicate that HDAC4 inhibition promotes CSC-derived cardiac regeneration and improves the restoration of cardiac function.
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Circulating microRNA-19a as a Potential Novel Biomarker for Diagnosis of Acute Myocardial Infarction.
Int J Mol Sci
PUBLISHED: 06-14-2014
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Acute myocardial infarction (AMI) is a serious cardiovascular disease. Investigating new susceptibility genes for effective methods of early diagnosis of AMI is important. In the current study, peripheral blood miR-19a levels were detected by real-time polymerase chain reaction. Significant differences and logistic correlation analyses were carried out by grouping of disease types and stratification of risk factors. Receiver-operator characteristic curve analysis was used to compare the current common clinical biochemical markers and evaluate the sensitivity and specificity of miR-19a for diagnosing AMI. Circulating miR-19a expression in the AMI group was higher than that in controls. The diagnostic effect of circulating miR-19a levels was superior to current clinical biochemical indices, such as CK, CK-MB, MYO, hs-TnI, and BNP. Our results show that there is a close association of circulating miR-19a levels with susceptibility to AMI. Circulating miR-19a levels could be a candidate diagnostic biomarker for AMI.
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Indian Hedgehog, a critical modulator in osteoarthritis, could be a potential therapeutic target for attenuating cartilage degeneration disease.
Connect. Tissue Res.
PUBLISHED: 06-13-2014
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The Hedgehog (Hh) family of proteins consists of Indian hedgehog (Ihh), sonic hedgehog (Shh), and desert hedgehog (Dhh). These proteins serve as essential regulators in a variety of developmental events. Ihh is mainly produced and secreted by prehypertrophic chondrocytes and regulates chondrocyte hypertrophy and endochondral bone formation during growth plate development. Tissue-specific deletion of the Ihh gene (targeted by Col2a1-Cre) causes early lethality in mice. Transgenic mice with induced Ihh expression exhibit increased chondrocyte hypertrophy and cartilage damage resembling human osteoarthritis (OA). During OA development, chondrocytes recapitulate the differentiation process that happens during the fetal status and which does not occur to an appreciable degree in adult articular cartilage. Ihh expression is up-regulated in human OA cartilage, and this upregulation correlates with OA progression and changes in chondrocyte morphology. A genetic study in mice further showed that conditional deletion of Ihh in chondrocytes attenuates OA progression, suggesting the possibility that blocking Ihh signaling can be used as a therapeutic approach to prevent or delay cartilage degeneration. However, Ihh gene deletion is currently not a therapeutic option as it is lethal in animals. RNA interference (RNAi) provides a means to knockdown Ihh without the severe side effects caused by chemical inhibitors. The currently available delivery methods for RNAi are nanoparticles and liposomes. Both have problems that need to be addressed. In the future, it will be necessary to develop a safe and effective RNAi delivery system to target Ihh signaling for preventing and treating OA.
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Targeting FoxM1 by thiostrepton inhibits growth and induces apoptosis of laryngeal squamous cell carcinoma.
J. Cancer Res. Clin. Oncol.
PUBLISHED: 06-08-2014
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We have previously reported that forkhead box M1 (FoxM1) transcription factor was overexpressed in laryngeal squamous cell carcinoma (LSCC) and was associated with development of LSCC. However, there are limited studies regarding the functional significance of FoxM1 and FoxM1 inhibitor thiostrepton in LSCC. Therefore, the aim of this study was to examine both in vitro and in vivo activity of FoxM1 inhibitor thiostrepton against LSCC cell line and nude mice.
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The upregulated expression of OX40/OX40L and their promotion of T cells proliferation in the murine model of asthma.
J Thorac Dis
PUBLISHED: 06-03-2014
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To investigate whether the expression of OX40/OX40 ligand (OX40L) was upregulated in a murine model of asthma and their significance in the pathogenesis of asthma.
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MicroRNA-1 regulates chondrocyte phenotype by repressing histone deacetylase 4 during growth plate development.
FASEB J.
PUBLISHED: 05-27-2014
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MicroRNAs (miRs) are noncoding RNAs (17-25 nt) that control translation and/or mRNA degradation. Using Northern blot analysis, we identified that miR-1 is specifically expressed in growth plate cartilage in addition to muscle tissue, but not in brain, intestine, liver, or lung. We obtained the first evidence that miR-1 is highly expressed in the hypertrophic zone of the growth plate, with an 8-fold increase compared with the proliferation zone; this location coincides with the Ihh and Col X expression regions in vivo. MiR-1 significantly induces chondrocyte proliferation and differentiation. We further identified histone deacetylase 4 (HDAC4) as a target of miR-1. HDAC4 negatively regulates chondrocyte hypertrophy by inhibiting Runx2, a critical transcription factor for chondrocyte hypertrophy. MiR-1 inhibits both endogenous HDAC4 protein by 2.2-fold and the activity of a reporter gene bearing the 3'-untranslated region (UTR) of HDAC4 by 3.3-fold. Conversely, knockdown of endogenous miR-1 relieves the repression of HDAC4. Deletion of the miR-1 binding site in HDAC4 3'-UTR or mutated miR-1 abolishes miR-1-mediated inhibition of the reporter gene activity. Overexpression of HDAC4 reverses miR-1 induction of chondrocyte differentiation markers Col X and Ihh. HDAC4 inhibits Runx2 promoter activity in a dosage-dependent manner. Thus, miR-1 plays an important role in the regulation of the chondrocyte phenotype during the growth plate development via direct targeting of HDAC4. - Li, P., Wei, X., Guan, Y., Chen, Q., Zhao, T., Sun, C., Wei, L. MicroRNA-1 regulates chondrocyte phenotype by repressing histone deacetylase 4 during growth plate development.
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Effects of emodin and irbesartan on ventricular fibrosis in Goldblatt hypertensive rats.
Pharmazie
PUBLISHED: 05-27-2014
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Left ventricular (LV) fibrosis is one of the most prominent pathophysiological results of hypertension. We initiated this study to investigate the effects and mechanisms of emodin and its combination with irbesartan on LV fibrosis in Goldblatt (2K1C) hypertensive rats. Goldblatt hypertension rats were prepared by two kidney one clip (2K1C) operations and then treated with either emodin, irbesartan or their combination. As a result, the systolic blood pressure (SBP) and the left ventricular mass index (LVMI) increased significantly (P < or = 0.05) in all 2K1C rats. After drugs treatment, irbesartan and the drug combination remarkably decreased SBP, LVMI, contents of angiotensinII (AngII), hydroxyproline and collagen, the mRNA and protein expression levels of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) (P < or = 0.05). As for the emodin, LVMI, contents of hydroxyproline and collagen, and MMP-2 and TIMP-2 expression were found to decrease significantly; however, the SBP and AngII contents stayed stable within certain extent. Therefore, emodin, irbesartan or two drugs together can potentially inhibit the ventricular fibrosis in Goldblatt hypertensive rats by reducing MMP-2 and TIMP-2 expression. Furthermore, the combination of these two drugs may provide a better anti-fibrosis effect than the single application.
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Preventive effect of crocin on osteoporosis in an ovariectomized rat model.
Evid Based Complement Alternat Med
PUBLISHED: 05-20-2014
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The purpose of this study was to investigate the therapeutic effects of crocin on ovariectomy-induced osteoporosis in rats. Female Sprague-Dawley rats were randomly assigned to a sham-operated group (sham) and five ovariectomy (OVX) subgroups, that is, OVX with vehicle (OVX), OVX with 17?-estradiol (E 2, 25??g/kg/day), and OVX with graded crocin doses (5, 10, or 20?mg/kg/day). Daily oral administration of E 2 or crocin started 4 weeks after OVX and lasted for 16 weeks. Our results showed that crocin dose-dependently inhibited the BMD reduction of L4 vertebrae and femurs caused by OVX and prevented the deterioration of trabecular microarchitecture, which were accompanied by a significant decrease in skeletal remodeling as evidenced by the lower levels of bone turnover markers. Furthermore, crocin reversed the oxidative stress status in both serum and bone tissue. The present study indicates that the administration of crocin at higher doses over a 16-week period can prevent OVX-induced osteoporosis in rats without hyperplastic effects on the uterus, which may, at least partially, be attributed to crocin's antioxidative property. In brief, crocin is a natural alternative for postmenopausal osteoporosis treatment in elderly women.
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Insulin improves osteogenesis of titanium implants under diabetic conditions by inhibiting reactive oxygen species overproduction via the PI3K-Akt pathway.
Biochimie
PUBLISHED: 05-13-2014
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Clinical evidence indicates that insulin therapy improves implant survival rates in diabetic patients; however, the mechanisms responsible for this effect are unknown. Here, we test if insulin exerts anti-oxidative effects, thereby improving diabetes-associated impaired osteoblast behavior on titanium implants. To test this hypothesis, we cultured primary rabbit osteoblasts in the presence of titanium implants and studied the impact of treatment with normal serum (NS), diabetic serum (DS), DS + insulin, DS + tempol (a superoxide dismutase mimetic), DS + insulin + tempol, and DS + insulin + wortmannin. We analyzed cell function, apoptosis, and reactive oxygen species (ROS) production in osteoblasts following the various treatments. Treatment with DS induced osteoblast dysfunction, evidenced by impaired cell attachment and morphology, decreased cell proliferation and ALP activity, and decreased expression of osteogenesis-related genes. We also observed a significant increase in apoptosis. Importantly, treatment with DS resulted in increased production of ROS in osteoblasts. In contrast, treatment with insulin inhibited ROS production, alleviated cell dysfunction, and decreased apoptosis of osteoblasts on the implants. Scavenging ROS with tempol also attenuated cell dysfunction. Compared to insulin treatment alone, the combination of insulin and tempol failed to further improve osteoblast functional recovery. Moreover, the anti-oxidative and pro-osteogenic effects afforded by insulin were almost completely abolished by the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin. These results demonstrate, for the first time, that insulin treatment alleviates the impaired osteogenesis of titanium implants under diabetic conditions by inhibiting ROS overproduction via a PI3K/Akt-dependent mechanism. Both the anti-oxidative and metabolic properties of insulin should make it a viable therapeutic option to combat diabetic implant failure.
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Caspase-8 mediates caspase-1 processing and innate immune defense in response to bacterial blockade of NF-?B and MAPK signaling.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 05-05-2014
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Toll-like receptor signaling and subsequent activation of NF-?B- and MAPK-dependent genes during infection play an important role in antimicrobial host defense. The YopJ protein of pathogenic Yersinia species inhibits NF-?B and MAPK signaling, resulting in blockade of NF-?B-dependent cytokine production and target cell death. Nevertheless, Yersinia infection induces inflammatory responses in vivo. Moreover, increasing the extent of YopJ-dependent cytotoxicity induced by Yersinia pestis and Yersinia pseudotuberculosis paradoxically leads to decreased virulence in vivo, suggesting that cell death promotes anti-Yersinia host defense. However, the specific pathways responsible for YopJ-induced cell death and how this cell death mediates immune defense against Yersinia remain poorly defined. YopJ activity induces processing of multiple caspases, including caspase-1, independently of inflammasome components or the adaptor protein ASC. Unexpectedly, caspase-1 activation in response to the activity of YopJ required caspase-8, receptor-interacting serine/threonine kinase 1 (RIPK1), and Fas-associated death domain (FADD), but not RIPK3. Furthermore, whereas RIPK3 deficiency did not affect YopJ-induced cell death or caspase-1 activation, deficiency of both RIPK3 and caspase-8 or FADD completely abrogated Yersinia-induced cell death and caspase-1 activation. Mice lacking RIPK3 and caspase-8 in their hematopoietic compartment showed extreme susceptibility to Yersinia and were deficient in monocyte and neutrophil-derived production of proinflammatory cytokines. Our data demonstrate for the first time to our knowledge that RIPK1, FADD, and caspase-8 are required for YopJ-induced cell death and caspase-1 activation and suggest that caspase-8-mediated cell death overrides blockade of immune signaling by YopJ to promote anti-Yersinia immune defense.
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Pressure-induced ferroelastic phase transition in SnO2 from density functional theory.
J Chem Phys
PUBLISHED: 05-03-2014
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High-pressure ferroelastic transition of rutile- to CaCl2-type SnO2 is investigated within density functional theory and Landau free energy theory. The calculated Landau energy map around the ground state is successfully used to clarify the softening mechanism of B1g mode (order parameter Q) and the coupling mechanism between the soft B1g mode and the soft transverse acoustic (TA) mode (strain ?). It is found that the Sn-O-Sn bending induced soft B1g mode effectively slows the excess energy increase caused by bond stretching, while the coupling between the soft B1g mode and the soft TA mode further decreases the energy since the lattice distortion strain ? minimizes the SnO6 octahedral distortion. Q induced Landau Gibbs free energy is interpreted as the sum of the bond stretching energy, bending energy, and octahedral distortion energy, while that induced by ? is interpreted as the lattice distortion energy.
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Antioxidant and hepatoprotective activity of Veronica ciliata Fisch. extracts against carbon tetrachloride-induced liver injury in mice.
Molecules
PUBLISHED: 04-10-2014
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Veronica ciliata Fisch. has been traditionally used in Traditional Chinese Medicine prescriptions due to its curative effects for hepatitis, cholecystitis, rheumatism, and urticaria. The present study was focused on investigating the role of ethyl acetate and aqueous extracts of Veronica ciliata Fisch. Furthermore, in vitro antioxidant activity (scavenging of DPPH, ABTS, superoxide, and nitrite radicals; reducing power; ?-carotene bleaching) and the hepatoprotective effect of the ethyl acetate extract by means of CCl4-induced oxidative stress in mice were investigated. The ethyl acetate extract of Veronica ciliata Fisch. displayed more noteworthy in vitro antioxidant activities than the aqueous extract. Moreover, it signi?cantly prevented the increase in serum T-AOC, ALT, AST and ALP level in acute liver damage induced by CCl4, decreased the extent of MDA formation in liver and elevated the activities of SOD and GSH in liver. This activity was found to be comparable to that of bifendate. Histopathological observation of the liver was also performed to further support the evidence from the biochemical analysis. The results indicated that strong antioxidant activities and a signi?cant protective effect against acute hepatotoxicity induced by CCl4 of Veronica ciliata Fisch. were concentrated in the ethyl acetate extract. The results suggested that this activity may be due to free radical-scavenging and antioxidant properties.
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The promotion of osteointegration under diabetic conditions using chitosan/hydroxyapatite composite coating on porous titanium surfaces.
Biomaterials
PUBLISHED: 04-01-2014
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Composited Chitosan/Hydroxyapatite (CS/HA) material coated on titanium surface (cTi) is a promising approach to produce biomaterials with better osseointegration capacity, but its bio-performance under diabetic conditions and the mechanisms involved remain elusive. We propose that the alterations in the Wnt/?-catenin pathway may play a role in mediating the improvement effect of cTi on diabetes-induced impaired implant osteointegration. To confirm the hypothesis, primary rat osteoblasts incubated on Ti and cTi were subjected to normal serum (NS), diabetic serum (DS), DS + Wnt3a (a specific Wnt agonist) and DS + Dkk1 (a specific Wnt antagonist) treatment. In vivo study was performed on diabetic sheep implanted with Ti or cTi into the bone defect on crista iliaca. Results showed that diabetes depressed osteoblast function evidenced by impaired cell adhesion and morphology, decreased cell proliferation and ALP activity, and higher apoptotic rate on Ti. Importantly, both cTi and Wnt3a treatment ameliorated osteoblastic dysfunction and apoptosis under diabetic condition. Implantation with cTi significantly improved osteointegration evidenced by Micro-CT and histological examinations compared with Ti. Moreover, the aforementioned promotive effects afforded by cTi were abolished by blocking Wnt pathway with Dkk1. Our study explicitly demonstrates that CS/HA composite material improves diabetes-induced impaired osteointegration of Ti via the reactivation of Wnt/?-catenin pathway and provides a target point for biomaterial modification to attain better clinical performance in diabetic patients.
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Crystallin ?B acts as a molecular guard in mouse decidualization: regulation and function during early pregnancy.
FEBS Lett.
PUBLISHED: 03-23-2014
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Although decidualization is crucial for the establishment of successful pregnancy, the molecular mechanism underlying decidualization remains poorly understood. Crystallin ?B (CryAB), a small heat shock protein (sHSP), is up-regulated and phosphorylated in mouse decidua. In mouse primary endometrial stromal cells, CryAB is induced upon progesterone treatment via HIF1?. In addition, CryAB is strongly phosphorylated through the p38-MAPK pathway under stress or during in vitro decidualization. Knockdown of CryAB results in the increase of apoptosis of stromal cells and inhibits decidualization under oxidative or inflammatory stress. Our data indicate that CryAB protects decidualization against stress conditions.
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Ritonavir binds to and downregulates estrogen receptors: molecular mechanism of promoting early atherosclerosis.
Exp. Cell Res.
PUBLISHED: 03-19-2014
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Estrogenic actions are closely related to cardiovascular disease. Ritonavir (RTV), a human immunodeficiency virus (HIV) protease inhibitor, induces atherosclerosis in an estrogen-related manner. However, how RTV induce pathological phenotypes through estrogen pathway remains unclear. In this study, we found that RTV increases thickness of coronary artery walls of Sprague Dawley rats and plasma free fatty acids (FFA) levels. In addition, RTV could induce foam cell formation, downregulate both estrogen receptor ? (ER?) and ER? expression, upregulate G protein-coupled estrogen receptor (GPER) expression, and all of them could be partially blocked by 17?-estradiol (E2), suggesting RTV acts as an antagonist for E2. Computational modeling shows a similar interaction with ER? between RTV and 2-aryl indoles, which are highly subtype-selective ligands for ER?. We also found that RTV directly bound to ER? and selectively inhibited the nuclear localization of ER?, and residue Leu536 in the hydrophobic core of ligand binding domain (LBD) was essential for the interaction with RTV. In addition, RTV did not change the secondary structure of ER?-LBD like E2, which explained how ER? lost the capacity of nuclear translocation under the treatment of RTV. All of the evidences suggest that ritonavir acts as an antagonist for 17?-estradiol in regulating ? subtype estrogen receptor function and early events of atherosclerosis.
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The effects of X-ray irradiation on the proliferation and apoptosis of MCF-7 breast cancer cells.
Ultrastruct Pathol
PUBLISHED: 03-13-2014
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To investigate the effects of X-ray irradiation on the proliferation and apoptosis of MCF-7 breast cancer cells; MCF-7 breast cancer cells were irradiated with X-ray. After irradiation, morphological changes and growth inhibition rate of the irradiated cells were observed under an inverted microscope. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to assess the proliferation of the irradiated MCF-7 cells. Transmission electron microscope was used to observe the morphology and ultrastructure of the irradiated MCF-7 cells. Western blotting was used to analyze the expression level of apoptosis-related protein caspase-3. Our results showed, at 48?h after the irradiation (0?Gy and 8?Gy), cells oval in shape, cell shrinkage or swelling and partial formation of debris under inverted microscope; as well as cytoplasmic vacuolization or inspissation, increased electron density of cytoplasm, structural damage of organelles, blurred mitochondrial cristae and chromatin margination under transmission electron microscopy; the survival rate of MCF-7 cells in X-ray group was 17.3% lower than that in control group (0?Gy) (p?
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The landscape of somatic mutations in epigenetic regulators across 1,000 paediatric cancer genomes.
Nat Commun
PUBLISHED: 03-12-2014
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Studies of paediatric cancers have shown a high frequency of mutation across epigenetic regulators. Here we sequence 633 genes, encoding the majority of known epigenetic regulatory proteins, in over 1,000 paediatric tumours to define the landscape of somatic mutations in epigenetic regulators in paediatric cancer. Our results demonstrate a marked variation in the frequency of gene mutations across 21 different paediatric cancer subtypes, with the highest frequency of mutations detected in high-grade gliomas, T-lineage acute lymphoblastic leukaemia and medulloblastoma, and a paucity of mutations in low-grade glioma and retinoblastoma. The most frequently mutated genes are H3F3A, PHF6, ATRX, KDM6A, SMARCA4, ASXL2, CREBBP, EZH2, MLL2, USP7, ASXL1, NSD2, SETD2, SMC1A and ZMYM3. We identify novel loss-of-function mutations in the ubiquitin-specific processing protease 7 (USP7) in paediatric leukaemia, which result in decreased deubiquitination activity. Collectively, our results help to define the landscape of mutations in epigenetic regulatory genes in paediatric cancer and yield a valuable new database for investigating the role of epigenetic dysregulations in cancer.
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The genomic landscape of diffuse intrinsic pontine glioma and pediatric non-brainstem high-grade glioma.
Nat. Genet.
PUBLISHED: 03-06-2014
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Pediatric high-grade glioma (HGG) is a devastating disease with a less than 20% survival rate 2 years after diagnosis. We analyzed 127 pediatric HGGs, including diffuse intrinsic pontine gliomas (DIPGs) and non-brainstem HGGs (NBS-HGGs), by whole-genome, whole-exome and/or transcriptome sequencing. We identified recurrent somatic mutations in ACVR1 exclusively in DIPGs (32%), in addition to previously reported frequent somatic mutations in histone H3 genes, TP53 and ATRX, in both DIPGs and NBS-HGGs. Structural variants generating fusion genes were found in 47% of DIPGs and NBS-HGGs, with recurrent fusions involving the neurotrophin receptor genes NTRK1, NTRK2 and NTRK3 in 40% of NBS-HGGs in infants. Mutations targeting receptor tyrosine kinase-RAS-PI3K signaling, histone modification or chromatin remodeling, and cell cycle regulation were found in 68%, 73% and 59% of pediatric HGGs, respectively, including in DIPGs and NBS-HGGs. This comprehensive analysis provides insights into the unique and shared pathways driving pediatric HGG within and outside the brainstem.
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Recurrent somatic structural variations contribute to tumorigenesis in pediatric osteosarcoma.
Cell Rep
PUBLISHED: 03-03-2014
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Pediatric osteosarcoma is characterized by multiple somatic chromosomal lesions, including structural variations (SVs) and copy number alterations (CNAs). To define the landscape of somatic mutations in pediatric osteosarcoma, we performed whole-genome sequencing of DNA from 20 osteosarcoma tumor samples and matched normal tissue in a discovery cohort, as well as 14 samples in a validation cohort. Single-nucleotide variations (SNVs) exhibited a pattern of localized hypermutation called kataegis in 50% of the tumors. We identified p53 pathway lesions in all tumors in the discovery cohort, nine of which were translocations in the first intron of the TP53 gene. Beyond TP53, the RB1, ATRX, and DLG2 genes showed recurrent somatic alterations in 29%-53% of the tumors. These data highlight the power of whole-genome sequencing for identifying recurrent somatic alterations in cancer genomes that may be missed using other methods.
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Regulation of Ku-DNA association by Yku70 C-terminal tail and SUMO modification.
J. Biol. Chem.
PUBLISHED: 02-24-2014
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The Ku70-Ku80 ring complex encloses DNA ends to facilitate telomere maintenance and DNA break repair. Many studies focus on the ring-forming regions of subunits Ku70 and Ku80. Less is known about the Ku70 C-terminal tail, which lies outside the ring. Our results suggest that this region is responsible for dynamic sumoylation of Yku70 upon DNA association in budding yeast. Mutating a cluster of five lysines in this region largely eliminates Yku70 sumoylation. Chromatin immunoprecipitation analyses show that yku70 mutants with these lysines replaced by arginines exhibit reduced Ku-DNA association at both telomeres and internal DNA breaks. Consistent with this physical evidence, Yku70 sumoylation deficiency is associated with impaired ability to block DNA end resection and suppression of multiple defects caused by inefficient resection. Correlating with these, yku70 mutants with reduced sumoylation levels exhibit shorter telomeres, increased G overhang levels, and altered levels of non-homologous end joining. We also show that diminution of sumoylation does not affect Yku70 protein levels or its interactions with protein and RNA partners. These results suggest a model whereby Yku70 sumoylation upon DNA association strengthens Ku-DNA interaction to promote multiple functions of Ku.
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Identification of ?2-macroglobulin as a master inhibitor of cartilage-degrading factors that attenuates the progression of posttraumatic osteoarthritis.
PUBLISHED: 02-20-2014
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To determine if supplemental intraarticular ?2-macroglobulin (?2 M) has a chondroprotective effect in a rat model of osteoarthritis (OA).
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Metabolomic analysis revealed that female mussel Mytilus galloprovincialis was sensitive to bisphenol A exposures.
Environ. Toxicol. Pharmacol.
PUBLISHED: 02-17-2014
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Bisphenol A (BPA) is a synthetic compound used in numerous chemicals, such as polycarbonate plastics and epoxy resins, and it can be released into aquatic environment and poses risk on aquatic organisms. In this work, metabolomics was applied to characterize the metabolic responses in mussel Mytilus galloprovincialis exposed to BPA. Our results indicated that the gonad of female mussel was sensitive to BPA exposures (1 and 10 ?g/L) for one month. However, no significant metabolic responses were observed in male mussel gonads exposed to these two concentrations of BPA. Overall, this limited study suggested that the gender differences should be considered in marine ecotoxicology.
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Indian hedgehog in synovial fluid is a novel marker for early cartilage lesions in human knee joint.
Int J Mol Sci
PUBLISHED: 02-10-2014
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To determine whether there is a correlation between the concentration of Indian hedgehog (Ihh) in synovial fluid (SF) and the severity of cartilage damage in the human knee joints, the knee cartilages from patients were classified using the Outer-bridge scoring system and graded using the Modified Mankin score. Expression of Ihh in cartilage and SF samples were analyzed with immunohistochemistry (IHC), western blot, and enzyme-linked immunosorbent assay (ELISA). Furthermore, we detected and compared Ihh protein levels in rat and mice cartilages between normal control and surgery-induced osteoarthritis (OA) group by IHC and fluorescence molecular tomography in vivo respectively. Ihh expression was increased 5.2-fold in OA cartilage, 3.1-fold in relative normal OA cartilage, and 1.71-fold in OA SF compared to normal control samples. The concentrations of Ihh in cartilage and SF samples was significantly increased in early-stage OA samples when compared to normal samples (r = 0.556; p < 0.001); however, there were no significant differences between normal samples and late-stage OA samples. Up-regulation of Ihh protein was also an early event in the surgery-induced OA models. Increased Ihh is associated with the severity of OA cartilage damage. Elevated Ihh content in human knee joint synovial fluid correlates with early cartilage lesions.
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Effect of bone material properties on effective region in screw-bone model: an experimental and finite element study.
Biomed Eng Online
PUBLISHED: 02-09-2014
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There have been numerous studies conducted to investigate the pullout force of pedicle screws in bone with different material properties. However, fewer studies have investigated the region of effect (RoE), stress distribution and contour pattern of the cancellous bone surrounding the pedicle screw.
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Direct renin inhibition with aliskiren protects against myocardial ischemia/reperfusion injury by activating nitric oxide synthase signaling in spontaneously hypertensive rats.
J Am Heart Assoc
PUBLISHED: 01-30-2014
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We tested the hypothesis that direct renin inhibition with aliskiren protects against myocardial ischemia/reperfusion (I/R) injury in spontaneously hypertensive rats (SHR), and examined the mechanism by which this occurs.
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Baicalin attenuates transforming growth factor-?1-induced human pulmonary artery smooth muscle cell proliferation and phenotypic switch by inhibiting hypoxia inducible factor-1? and aryl hydrocarbon receptor expression.
J. Pharm. Pharmacol.
PUBLISHED: 01-23-2014
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Baicalin, a natural flavone, has antithrombotic, antihyperlipidemic and antiinflammortory activity. It can also inhibit cancer cell proliferation and reduce brain cell apoptosis. This study aimed to elucidate the effect of baicalin on the excessive proliferation of human pulmonary arterial smooth muscle cells (HPASMCs) induced by transforming growth factor-?1 (TGF-?1) and to investigate the roles of hypoxia inducible factor-1? (HIF-1?) and aryl hydrocarbon receptor (AhR) in mediating this TGF-?1-induced excessive proliferation of HPASMCs.
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Finite element analysis of a bone healing model: 1-year follow-up after internal fixation surgery for femoral fracture.
Pak J Med Sci
PUBLISHED: 01-18-2014
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Finite element analysis was used to compare preoperative and postoperative stress distribution of a bone healing model of femur fracture, to identify whether broken ends of fractured bone would break or not after fixation dislodgement one year after intramedullary nailing. Method s: Using fast, personalized imaging, bone healing models of femur fracture were constructed based on data from multi-slice spiral computed tomography using Mimics, Geomagic Studio, and Abaqus software packages. The intramedullary pin was removed by Boolean operations before fixation was dislodged. Loads were applied on each model to simulate a person standing on one leg. The von Mises stress distribution, maximum stress, and its location was observed. Results : According to 10 kinds of display groups based on material assignment, the nodes of maximum and minimum von Mises stress were the same before and after dislodgement, and all nodes of maximum von Mises stress were outside the fracture line. The maximum von Mises stress node was situated at the bottom quarter of the femur. The von Mises stress distribution was identical before and after surgery. Conclusion : Fast, personalized model establishment can simulate fixation dislodgement before operation, and personalized finite element analysis was performed to successfully predict whether nail dislodgement would disrupt femur fracture or not.
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Expansion on a matrix deposited by nonchondrogenic urine stem cells strengthens the chondrogenic capacity of repeated-passage bone marrow stromal cells.
Cell Tissue Res.
PUBLISHED: 01-09-2014
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Human urine-derived stem cells (hUSCs) are a newly found type of stem cell with a potential for therapeutic application in urology. The aim of this study is to investigate whether hUSCs contribute to cartilage regeneration. Despite their characterization with multi-lineage differentiation capacities, in terms of osteogenesis, adipogenesis and myogenesis, hUSCs do not show the ability to differentiate into chondrocytes. Human bone marrow stromal cells (hBMSCs) are a tissue-specific stem cell for endochondral bone formation; however, repeated-passage hBMSCs have a lower capacity for chondrogenic differentiation. We found that the extracellular matrix (ECM) deposited by hUSCs (UECM) can greatly recharge repeated-passage hBMSCs toward chondrogenic differentiation, a result that might be explained by trophic factors released from hUSCs being immobilized in UECM. We also found that ECM from repeated-passage hBMSCs (BECM) have a limited rejuvenation effect. The Wnt11-mediated noncanonical signaling pathway might be responsible for UECM-mediated hBMSC rejuvenation and subsequent chondrogenic differentiation. Our data indicate that commercially available UECM from young healthy donors might represent a simple and promising approach for autologous hBMSC rejuvenation. This study also provides an excellent model for investigating the effect of trophic factors released by stem cells on tissue regeneration without interference by stem cell differentiation.
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Disrupting the Indian hedgehog signaling pathway in vivo attenuates surgically induced osteoarthritis progression in Col2a1-CreERT2; Ihhfl/fl mice.
Arthritis Res. Ther.
PUBLISHED: 01-08-2014
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Previous observations implicate Indian hedgehog (Ihh) signaling in osteoarthritis (OA) development because it regulates chondrocyte hypertrophy and matrix metallopeptidase 13 (MMP-13) expression. However, there is no direct genetic evidence for the role of Ihh in OA, because mice with cartilage or other tissue-specific deletion of the Ihh gene die shortly after birth. We evaluated the role of Ihh in vivo via a Cre-loxP-mediated approach to circumvent the early death caused by Ihh deficiency.
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TNF-? mediated increase of HIF-1? inhibits VASP expression, which reduces alveolar-capillary barrier function during acute lung injury (ALI).
PLoS ONE
PUBLISHED: 01-01-2014
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Acute lung injury (ALI) is an inflammatory disorder associated with reduced alveolar-capillary barrier function and increased pulmonary vascular permeability. Vasodilator-stimulated phosphoprotein (VASP) is widely associated with all types of modulations of cytoskeleton rearrangement-dependent cellular morphology and function, such as adhesion, shrinkage, and permeability. The present studies were conducted to investigate the effects and mechanisms by which tumor necrosis factor-alpha (TNF-?) increases the tight junction permeability in lung tissue associated with acute lung inflammation. After incubating A549 cells for 24 hours with different concentrations (0-100 ng/mL) of TNF-?, 0.1 to 8 ng/mL TNF-? exhibited no significant effect on cell viability compared with the 0 ng/mL TNF-? group (control group). However, 10 ng/mL and 100 ng/mL TNF-? dramatically inhibited the viability of A549 cells compared with the control group (*p<0.05). Monolayer cell permeability assay results indicated that A549 cells incubated with 10 ng/mL TNF-? for 24 hours displayed significantly increased cell permeability (*p<0.05). Moreover, the inhibition of VASP expression increased the cell permeability (*p<0.05). Pretreating A549 cells with cobalt chloride (to mimic a hypoxia environment) increased protein expression level of hypoxia inducible factor-1? (HIF-1?) (*p<0.05), whereas protein expression level of VASP decreased significantly (*p<0.05). In LPS-induced ALI mice, the concentrations of TNF-? in lung tissues and serum significantly increased at one hour, and the value reached a peak at four hours. Moreover, the Evans Blue absorption value of the mouse lung tissues reached a peak at four hours. The HIF-1? protein expression level in mouse lung tissues increased significantly at four hours and eight hours (**p<0.001), whereas the VASP protein expression level decreased significantly (**p<0.01). Taken together, our data demonstrate that HIF-1? acts downstream of TNF-? to inhibit VASP expression and to modulate the acute pulmonary inflammation process, and these molecules play an important role in the impairment of the alveolar-capillary barrier.
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ROCK1 deficiency enhances protective effects of antioxidants against apoptosis and cell detachment.
PLoS ONE
PUBLISHED: 01-01-2014
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We have recently reported that the homologous Rho kinases, ROCK1 and ROCK2, play different roles in regulating stress-induced stress fiber disassembly and cell detachment, and the ROCK1 deficiency in mouse embryonic fibroblasts (MEF) has remarkable anti-apoptotic, anti-detachment and pro-survival effects against doxorubicin, a chemotherapeutic drug. This study investigated the roles of ROCK isoforms in doxorubicin-induced reactive oxygen species (ROS) generation which is believed to be the major mechanism underlying its cytotoxicity to normal cells, and especially to cardiomyocytes. Different antioxidants have been shown to provide a protective role reported in numerous experimental studies, but clinical trials of antioxidant therapy showed insufficient benefit against the cardiac side effect. We found that both ROCK1-/- and ROCK2-/- MEFs exhibited reduced ROS production in response to doxorubicin treatment. Interestingly, only ROCK1 deficiency, but not ROCK2 deficiency, significantly enhanced the protective effects of antioxidants against doxorubicin-induced cytotoxicity. First, ROCK1 deficiency and N-acetylcysteine (an anti-oxidant) treatment synergistically reduced ROS levels, caspase activation and cell detachment. In addition, the reduction of ROS generation in ROCK1-/- MEFs in response to doxorubicin treatment was in part through inhibiting NADPH oxidase activity. Furthermore, ROCK1 deficiency enhanced the inhibitory effects of diphenyleneiodonium (an inhibitor of NADPH oxidase) on ROS generation and caspase 3 activation induced by doxorubicin. Finally, ROCK1 deficiency had greater protective effects than antioxidant treatment, especially on reducing actin cytoskeleton remodeling. ROCK1 deficiency not only reduced actomyosin contraction but also preserved central stress fiber stability, whereas antioxidant treatment only reduced actomyosin contraction without preserving central stress fibers. These results reveal a novel strategy to enhance the protective effect of antioxidant therapy by targeting the ROCK1 pathway to stabilize the actin cytoskeleton and boost the inhibitory effects on ROS production, apoptosis and cell detachment.
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Azithromycin inhibits double-stranded RNA-induced thymic stromal lymphopoietin release from human airway epithelial cells.
Pharmazie
PUBLISHED: 12-31-2013
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Thymic stromal lymphopoietin (TSLP) is elevated in asthma and triggers dendritic cell-mediated activation of TH2 inflammatory responses. Viral stimuli, a major cause of asthma exacerbations, have been shown to induce overexpression of TSLP in asthmatic epithelium. Azithromycin has various anti-microbial and antiinflammatory effects. However, the effect of azithromycin on the production of TSLP has not been studied. Here we explored the effects of azithromycin on viral surrogate (dsRNA)-induced TSLP in normal human bronchial epithelial (NHBE) cells. NHBE were stimulated with poly (I:C) in the presence azithromycin. The effects of azithromycin on dsRNA-induced inflammatory responses in NHBE cells were analyzed. We demonstrated that azithromycin inhibited the production and mRNA expression of TSLP in NHBE cells. Azithromycin also inhibited the nuclear factor-KB luciferase activity induced by poly (I:C), and it prevented dsRNA-induced loss of the NF-kappaB repressor protein IkappaBalpha. These results suggest that azithromycin can be useful to treat asthma exacerbations due to the inhibition of TSLP.
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ROCK1 isoform-specific deletion reveals a role for diet-induced insulin resistance.
Am. J. Physiol. Endocrinol. Metab.
PUBLISHED: 12-10-2013
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Rho-kinase (ROCK) isoforms regulate insulin signaling and glucose metabolism negatively or positively in cultured cell lines and skeletal muscle. However, the in vivo function of the ROCK1 isoform in adipose tissue has not been addressed. To determine the specific role of the adipose ROCK1 isoform in the development of insulin resistance and obesity, mice lacking ROCK1 in adipose tissue globally or selectively were studied. Here we show that insulins ability to activate IRS-1/PI3K/Akt signaling was greatly enhanced in adipose tissue of ROCK1-/- mice compared with wild type mice. These effects result from the inhibitory effect of ROCK1 on IR action, as evidenced by the fact that IR tyrosine phosphorylation was abolished in ROCK1-/- MEF cells when ROCK1 was reexpressed. Consistently, adipose-specific disruption of ROCK1 increased IR tyrosine phosphorylation in adipose tissue and modestly improved sensitivity to insulin in obese mice induced by high-fat feeding. This effect is independent of any changes in adiposity, number or size of adipocytes, and metabolic parameters, including glucose, insulin, leptin, and TG levels, demonstrating a minimal effect of adipose ROCK1 on whole-body metabolism. Enzymatic activity of ROCK1 in adipose tissue remained ~50%, which likely originated from the fraction of stromal vascular cells, suggesting involvement of these cells for adipose metabolic regulation. Moreover, ROCK isoform activities were increased in adipose tissue of diet-induced or genetically obese mice. These data suggest that adipose ROCK1 isoform plays an inhibtory role for the regulation of insulin sensitivity in diet-induced obesity in vivo.
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Features of urate deposition in patients with gouty arthritis of the foot using dual-energy computed tomography.
Int J Rheum Dis
PUBLISHED: 11-19-2013
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To investigate features of urate deposition in gout and the association between these features and attacks of gouty arthritis using dual-energy computed tomography (CT).
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[Biomechanical study on effects of bone mineral density on fixation strength of expansive pedicle screw].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
PUBLISHED: 11-01-2013
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To evaluate the fixation strength of expansive pedicle screw (EPS) at different bone mineral density (BMD) levels, further to provide theoretical evidence for the clinical application of the EPS in patients with osteoporosis.
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[Three-dimensional flow perfusion culture enhances proliferation of human fetal osteoblasts in large scaffold with controlled architecture].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 10-31-2013
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To demonstrate the feasibility and benefits of custom designed perfusion bioreactor in conjunction with well-defined three-dimensional (3D) environment for enhanced proliferation and homogeneous distribution of human fetal osteoblasts in large scaffold in vitro.
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Effect of Preoperative Tracheal Stretch Exercise on Anterior Cervical Spine Surgery: A Retrospective Study.
J Spinal Disord Tech
PUBLISHED: 10-19-2013
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We designed a retrospective study of preoperative tracheal stretch exercise (TSE) before anterior cervical spine surgery. The changes of vital signs before and during the surgery and the postoperative clinical outcome were recorded and compared with none treated patients.
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ZnO electron field emitters on three-dimensional patterned carbon nanotube framework.
ACS Appl Mater Interfaces
PUBLISHED: 09-04-2013
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A three-dimensional patterned CNT framework was prepared in a thermal chemical vapor deposition system. ZnO nanoneedles with different areal density were subsequently grown on it via hydrothermal method. By combining the advantages of high aspect ratio, more effective emission sites with minimized screen effect and good Ohmic contact between ZnO nanoneedles and CNT framework, the ZnO/CNT hierarchical nanostructures with medium areal density exhibit a favorable FE performance which makes it promising candidate for cold cathode nanomaterials.
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Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.
Proteomics
PUBLISHED: 08-25-2013
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In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.
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The Protective Effect of Cordymin, a Peptide Purified from the Medicinal Mushroom Cordyceps sinensis, on Diabetic Osteopenia in Alloxan-Induced Diabetic Rats.
Evid Based Complement Alternat Med
PUBLISHED: 08-17-2013
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The aim of this study was to investigate the protective effect of cordymin on diabetic osteopenia in alloxan-induced diabetic rats and the possible mechanisms involved. The diabetic rats received daily intraperitoneal injection with cordymin (20, 50, and 100?mg/kg/day) for 5 weeks. Cordymin could restore the circulating blood glucose, glycosylated hemoglobin (HbA1c), serum alkaline phosphatase (ALP), tartrate resistant acid phosphatase (TRAP), and insulin levels in a dose-dependent manner. Also, the treatment of diabetic rats with cordymin could partially reverse the ? cells death and decrease the total antioxidant status (TAOS) in the diabetic rats. The results may directly and indirectly account for the possible mechanism of the beneficial effect of cordymin on diabetic osteopenia, which was confirmed with the increased bone mineral content (BMC) and bone mineral density (BMD) in diabetic rats (P < 0.05). All those findings indicate that cordymin may play a protective role in diabetic osteoporosis.
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Rho kinases in cardiovascular physiology and pathophysiology: the effect of fasudil.
J. Cardiovasc. Pharmacol.
PUBLISHED: 08-08-2013
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Rho kinase (ROCK) is a major downstream effector of the small GTPase RhoA. ROCK family, consisting of ROCK1 and ROCK2, plays central roles in the organization of actin cytoskeleton and is involved in a wide range of fundamental cellular functions, such as contraction, adhesion, migration, proliferation, and apoptosis. Due to the discovery of effective inhibitors, such as fasudil and Y27632, the biological roles of ROCK have been extensively explored with particular attention on the cardiovascular system. In many preclinical models of cardiovascular diseases, including vasospasm, arteriosclerosis, hypertension, pulmonary hypertension, stroke, ischemia-reperfusion injury, and heart failure, ROCK inhibitors have shown a remarkable efficacy in reducing vascular smooth muscle cell hypercontraction, endothelial dysfunction, inflammatory cell recruitment, vascular remodeling, and cardiac remodeling. Moreover, fasudil has been used in the clinical trials of several cardiovascular diseases. The continuing utilization of available pharmacological inhibitors and the development of more potent or isoform-selective inhibitors in ROCK signaling research and in treating human diseases are escalating. In this review, we discuss the recent molecular, cellular, animal, and clinical studies with a focus on the current understanding of ROCK signaling in cardiovascular physiology and diseases. We particularly note that emerging evidence suggests that selective targeting ROCK isoform based on the disease pathophysiology may represent a novel therapeutic approach for the disease treatment including cardiovascular diseases.
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Advanced understanding of stickiness on superhydrophobic surfaces.
Sci Rep
PUBLISHED: 07-11-2013
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This study explores how contact angle hysteresis and titling angle relate with stickiness on superhydrophobic surfaces. The result indicates that contact angle hysteresis could not be mentioned as a proper factor to evaluate the surface stickiness. By analyzing the system pinning force of droplet placed on a titled surface, we concluded that both solid fraction and surface geometric factor are the critical factors determining the surface stickiness.
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Proteomic and metabolomic responses in hepatopancreas of Mytilus galloprovincialis challenged by Micrococcus luteus and Vibrio anguillarum.
J Proteomics
PUBLISHED: 07-09-2013
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The outbreak of pathogens can induce diseases and lead to massive mortalities of aquaculture animals including fish, mollusk and shrimp. In this work, the responses induced by Micrococcus luteus and Vibrio anguillarum were investigated in hepatopancreas of mussel Mytilus galloprovincialis using proteomics and metabolomics. Metabolic biomarkers demonstrated that M. luteus and V. anguillarum injections could induce osmotic stress and disturbance in energy metabolism. And the uniquely and more markedly altered metabolic biomarkers (glutamine, succinate, aspartate, glucose, ATP, homarine and tyrosine) indicated that V. anguillarum could cause more severe disturbances in osmotic regulation and energy metabolism. The differentially altered proteins meant that M. luteus and V. anguillarum induced different effects in mussels. However, the common proteomic biomarkers, arginine kinase and small heat shock protein, demonstrated that these two bacteria induced similar effects including oxidative stress and disturbance in energy metabolism in M. galloprovincialis. In addition, some metabolic biomarkers, ATP and glutamine, were confirmed by related proteins including arginine kinase, ATP synthase, nucleoside diphosphate kinase and glutamine synthetase in bacteria-challenged mussels. This study demonstrated that proteomics and metabolomics could provide an insightful view into the effects of environmental pathogens to the marine mussel M. galloprovincialis.
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Ouabain-induced apoptosis in cochlear hair cells and spiral ganglion neurons in vitro.
Biomed Res Int
PUBLISHED: 07-04-2013
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Ouabain is a common tool to explore the pathophysiological changes in adult mammalian cochlea in vivo. In prior studies, locally administering ouabain via round window membrane demonstrated that the ototoxic effects of ouabain in vivo varied among mammalian species. Little is known about the ototoxic effects in vitro. Thus, we prepared cochlear organotypic cultures from postnatal day-3 rats and treated these cultures with ouabain at 50, 500, and 1000 ?M for different time to elucidate the ototoxic effects of ouabain in vitro and to provide insights that could explain the comparative ototoxic effects of ouabain in vivo. Degeneration of cochlear hair cells and spiral ganglion neurons was evaluated by hair-cell staining and neurofilament labeling, respectively. Annexin V staining was used to detect apoptotic cells. A quantitative RT-PCR apoptosis-focused gene array determined changes in apoptosis-related genes. The results showed that ouabain-induced damage in vitro was dose and time dependent. 500 ?M ouabain and 1000 ?M ouabain were destructively traumatic to both spiral ganglion neurons and cochlear hair cells in an apoptotic signal-dependent pathway. The major apoptotic pathways in ouabain-induced spiral ganglion neuron apoptosis culminated in the stimulation of the p53 pathway and triggering of apoptosis by a network of proapoptotic signaling pathways.
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Cross-species genomic and epigenomic landscape of retinoblastoma.
Oncotarget
PUBLISHED: 06-15-2013
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Genetically engineered mouse models (GEMMs) of human cancer are important for advancing our understanding of tumor initiation and progression as well as for testing novel therapeutics. Retinoblastoma is a childhood cancer of the developing retina that initiates with biallelic inactivation of the RB1 gene. GEMMs faithfully recapitulate the histopathology, molecular, cellular, morphometric, neuroanatomical and neurochemical features of human retinoblastoma. In this study, we analyzed the genomic and epigenomic landscape of murine retinoblastoma and compared them to human retinoblastomas to gain insight into shared mechanisms of tumor progression across species. Similar to human retinoblastoma, mouse tumors have low rates of single nucleotide variations. However, mouse retinoblastomas have higher rates of aneuploidy and regional and focal copy number changes that vary depending on the genetic lesions that initiate tumorigenesis in the developing murine retina. Furthermore, the epigenetic landscape in mouse retinoblastoma was significantly different from human tumors and some pathways that are candidates for molecular targeted therapy for human retinoblastoma such as SYK or MCL1 are not deregulated in GEMMs. Taken together, these data suggest there are important differences between mouse and human retinoblastomas with respect to the mechanism of tumor progression and those differences can have significant implications for translational research to test the efficacy of novel therapies for this devastating childhood cancer.
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CXCR4-targeted therapy inhibits VEGF expression and chondrosarcoma angiogenesis and metastasis.
Mol. Cancer Ther.
PUBLISHED: 05-17-2013
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Chondrosarcoma is notable for its lack of response to conventional cytotoxic chemotherapy, propensity for developing lung metastases, and poor survival. Therefore, a better understanding of angiogenic and metastatic pathways is needed. Multiple pathways regulate angiogenesis and metastasis, including chemokines and their receptors. In this study, we investigated chemokine (C-X-C motif) receptor 4 (CXCR4) signaling in chondrosarcoma and tested the hypotheses that CXCR4 inhibition suppresses tumor angiogenesis and metastasis. CXCR4 expression, analyzed by real-time PCR and Western blot, was increased in human chondrosarcoma cell line JJ compared with normal chondrocytes and was further increased in JJ by hypoxia (2% O2), vascular endothelial growth factor A (VEGFA; 10 ng/mL), and in xenograft tumors in nude mice. The CXCR4 ligand CXCL12 (10 ng/mL) doubled secreted VEGFA, measured with ELISA, under hypoxic conditions and this conditioned media increased human umbilical vein endothelial cell tube formation. These effects were inhibited by CXCR4 siRNA or AMD3100 (5 ?g/mL). In a xenograft mouse model, four weeks of AMD3100 treatment (1.25 mg/kg, intraperitoneally twice daily) inhibited tumor angiogenesis, tumor growth, and metastasis. VEGFA content in tumor extracts was decreased (7.19 ± 0.52 ng/mL control vs. 3.96 ± 0.66 treatment) and bioimaging of angiogenesis was decreased by 56%. Tumor volumes averaged 4.44 ± 0.68 cm(3) in control compared with 2.48 ± 0.61 cm(3) in the treatment group. The number of lung metastatic nodules was 23 ± 9 in control compared with 10 ± 6 in the treatment group (N = 8/group). Therefore, CXCR4-targeted therapy may be a treatment strategy for chondrosarcoma.
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Proteomic and metabolomic analysis of earthworm Eisenia fetida exposed to different concentrations of 2,2,4,4-tetrabromodiphenyl ether.
J Proteomics
PUBLISHED: 05-10-2013
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As a class of brominated flame-retardants (BFRs), polybrominated diphenyl ethers (PBDEs) are widely used in industrial products. PBDEs have been detected in local terrestrial biota from the Laizhou Bay in China. They can induce various toxicities, such as hepatotoxicity, neurotoxicity, cytotoxicity, genotoxicity and endocrine disrupting effects in animals. In this work, we characterized the dose-responsive effects of 2,2,4,4-tetrabromodiphenyl ether (BDE 47) in earthworm Eisenia fetida using an integrated proteomic and metabolomic approach. Metabolic responses indicated that BDE 47 mainly caused disturbance in osmotic regulation and energy metabolism marked by differentially altered betaine, amino acids, ATP, glucose, maltose and succinate in E. fetida. Proteomic responses revealed that BDE 47 induced cell apoptosis (or injury), oxidative stress, disturbance in protein biosynthesis and energy metabolism in E. fetida in terms of differential proteomic biomarkers. Especially, the increased ATP was confirmed by up-regulated nucleoside diphosphate kinase A and ATP synthase in 1 and 100?g/L of BDE 47-treated groups, respectively. In addition, several metabolic biomarkers including betaine, glycine and 2-hexyl-5-ethyl-3-furansulfonate were relatively stable in all BDE 47-exposed groups. This work demonstrated that proteomics and metabolomics could partially validate one another and their combination could better understand toxicological effects of environmental pollutants.
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Targeting oxidative stress in embryonal rhabdomyosarcoma.
Cancer Cell
PUBLISHED: 05-03-2013
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Rhabdomyosarcoma is a soft-tissue sarcoma with molecular and cellular features of developing skeletal muscle. Rhabdomyosarcoma has two major histologic subtypes, embryonal and alveolar, each with distinct clinical, molecular, and genetic features. Genomic analysis shows that embryonal tumors have more structural and copy number variations than alveolar tumors. Mutations in the RAS/NF1 pathway are significantly associated with intermediate- and high-risk embryonal rhabdomyosarcomas (ERMS). In contrast, alveolar rhabdomyosarcomas (ARMS) have fewer genetic lesions overall and no known recurrently mutated cancer consensus genes. To identify therapeutics for ERMS, we developed and characterized orthotopic xenografts of tumors that were sequenced in our study. High-throughput screening of primary cultures derived from those xenografts identified oxidative stress as a pathway of therapeutic relevance for ERMS.
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Proteomic and metabolomic analysis reveal gender-specific responses of mussel Mytilus galloprovincialis to 2,2,4,4-tetrabromodiphenyl ether (BDE 47).
Aquat. Toxicol.
PUBLISHED: 04-27-2013
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Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame-retardants (BFRs) that are widely used in industrial products and have posed potential risk on the coastal environment of the Laizhou Bay in China. They are of great concern due to their toxicities, such as hepatotoxicity, carcinogenecity, neurotoxicity, immunotoxicity and endocrine disrupting effects in animals. In this work, we focused on the gender-specific responses of BDE 47 in mussel Mytilus galloprovincialis using a combined proteomic and metabolomic approach. Metabolic responses indicated that BDE 47 mainly caused disturbance in energy metabolism in male mussel gills. For female mussel samples, disruption in both osmotic regulation and energy metabolism was found in terms of differential metabolic profiles. Proteomic responses revealed that BDE 47 induced cell apoptosis and reduced reactive oxygen species (ROS) production in both male and female mussels, disturbance in protein homeostasis in male mussels as well as disturbance in female mussel proteolysis based on the differential proteomic biomarkers. Overall, these results confirmed the gender-specific responses in mussels to BDE 47 exposures. This work demonstrated that an integrated metabolomic and proteomic approach could provide an important insight into the toxicological effects of environmental pollutant to organisms.
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Anticataleptic effects of 5-HT1B receptors in the globus pallidus.
Neurosci. Res.
PUBLISHED: 04-21-2013
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The globus pallidus occupies an important position in the indirect pathway of the basal ganglia. Being a monoamine neurotransmitter, 5-HT is involved in mediating many physiological functions and pathophysiological processes in several movement disorders. Morphological studies have revealed that the globus pallidus receives serotonergic innervation arising from the raphe nuclei, mainly the dorsal raphe nucleus. A high level of 5-HT and 5-HT1B receptors were detected in the globus pallidus. In the present study, bilateral microinjection of 5-HT or 5-HT1B receptor agonist, CP-93129, into the globus pallidus significantly alleviated the symptoms of rigidity caused by haloperidol. To further elucidate 5-HT1B receptor-induced anticatalepsy, in vivo extracellular recordings were performed to examine the effects of 5-HT1B receptor activation on the firing activity of the globus pallidus neurons under the presence of haloperidol. Micro-pressure ejection of 5-HT or CP-93129 increased the spontaneous firing rate of the pallidal neurons. Furthermore, by using immunohistochemistry, positive staining of 5-HT1B receptor was observed in the globus pallidus neurons. Taken together, the present findings provide evidence that activation of 5-HT1B receptor may exert anticataleptic effects by increasing the activity of pallidal neurons.
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Dissecting the roles of ROCK isoforms in stress-induced cell detachment.
Cell Cycle
PUBLISHED: 04-18-2013
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The homologous Rho kinases, ROCK1 and ROCK2, are involved in stress fiber assembly and cell adhesion and are assumed to be functionally redundant. Using mouse embryonic fibroblasts (MEFs) derived from ROCK1(-/-) and ROCK2(-/-) mice, we have recently reported that they play different roles in regulating doxorubicin-induced stress fiber disassembly and cell detachment: ROCK1 is involved in destabilizing the actin cytoskeleton and cell detachment, whereas ROCK2 is required for stabilizing the actin cytoskeleton and cell adhesion. Here, we present additional insights into the roles of ROCK1 and ROCK2 in regulating stress-induced impairment of cell-matrix and cell-cell adhesion. In response to doxorubicin, ROCK1(-/-) MEFs showed significant preservation of both focal adhesions and adherens junctions, while ROCK2(-/-) MEFs exhibited impaired focal adhesions but preserved adherens junctions compared with the wild-type MEFs. Additionally, inhibition of focal adhesion or adherens junction formations by chemical inhibitors abolished the anti-detachment effects of ROCK1 deletion. Finally, ROCK1(-/-) MEFs, but not ROCK2(-/-) MEFs, also exhibited preserved central stress fibers and reduced cell detachment in response to serum starvation. These results add new insights into a novel mechanism underlying the anti-detachment effects of ROCK1 deletion mediated by reduced peripheral actomyosin contraction and increased actin stabilization to promote cell-cell and cell-matrix adhesion. Our studies further support the differential roles of ROCK isoforms in regulating stress-induced loss of central stress fibers and focal adhesions as well as cell detachment.
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The age-related changes in cartilage and osteoarthritis.
Biomed Res Int
PUBLISHED: 04-03-2013
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Osteoarthritis (OA) is closely associated with aging, but its underlying mechanism is unclear. Recent publications were reviewed to elucidate the connection between aging and OA. With increasing OA incidence, more senior people are facing heavy financial and social burdens. Age-related OA pathogenesis is not well understood. Recently, it has been realized that age-related changes in other tissues besides articular cartilage may also contribute to OA development. Many factors including senescence-related secretory phenotypes, chondrocytes low reactivity to growth factors, mitochondrial dysfunction and oxidative stress, and abnormal accumulation of advanced glycation end products (AGEs) may all play key roles in the pathogenesis of age-related OA. Lately, epigenetic regulation of gene expression was recognized for its impact on age-related OA pathogenesis. Up to now, few studies have been reported about the role of miRNA and long-noncoding RNA (lncRNA) in age-related OA. Research focusing on this area may provide valuable insights into OA pathogenesis. OA-induced financial and social burdens have become an increasingly severe threat to older population. Age-related changes in noncartilage tissue should be incorporated in the understanding of OA development. Growing attention on oxidative stress and epigenetics will provide more important clues for the better understanding of the age-related OA.
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Global chromatin profiling reveals NSD2 mutations in pediatric acute lymphoblastic leukemia.
Nat. Genet.
PUBLISHED: 03-29-2013
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Epigenetic dysregulation is an emerging hallmark of cancers. We developed a high-information-content mass spectrometry approach to profile global histone modifications in human cancers. When applied to 115 lines from the Cancer Cell Line Encyclopedia, this approach identified distinct molecular chromatin signatures. One signature was characterized by increased histone 3 lysine 36 (H3K36) dimethylation, exhibited by several lines harboring translocations in NSD2, which encodes a methyltransferase. A previously unknown NSD2 p.Glu1099Lys (p.E1099K) variant was identified in nontranslocated acute lymphoblastic leukemia (ALL) cell lines sharing this signature. Ectopic expression of the variant induced a chromatin signature characteristic of NSD2 hyperactivation and promoted transformation. NSD2 knockdown selectively inhibited the proliferation of NSD2-mutant lines and impaired the in vivo growth of an NSD2-mutant ALL xenograft. Sequencing analysis of >1,000 pediatric cancer genomes identified the NSD2 p.E1099K alteration in 14% of t(12;21) ETV6-RUNX1-containing ALLs. These findings identify NSD2 as a potential therapeutic target for pediatric ALL and provide a general framework for the functional annotation of cancer epigenomes.
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Responses of Mytilus galloprovincialis to bacterial challenges by metabolomics and proteomics.
Fish Shellfish Immunol.
PUBLISHED: 03-28-2013
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Pathogens can cause diseases and lead to massive mortalities of aquaculture animals and substantial economic loss. In this work, we studied the responses induced by Micrococcus luteus and Vibrio anguillarum in gill of mussel Mytilus galloprovincialis at protein and metabolite levels. Metabolic biomarkers (e.g., amino acids, betaine, ATP) suggested that both M. luteus and V. anguillarum induced disturbances in energy metabolism and osmotic regulation. The unique and some more remarkably altered metabolic biomarkers (threonine, alanine, aspartate, taurine, succinate) demonstrated that V. anguillarum could cause more severe disturbances in osmotic regulation and energy metabolism. Proteomic biomarkers (e.g., goose-type lysozyme 2, matrilin, ependymin-related protein, peptidyl-prolyl cis-trans isomerases) indicated that M. luteus caused immune stress, and disturbances in signaling pathways and protein synthesis. However, V. anguillarum mainly induced oxidative stress and disturbance in energy metabolism in mussel gills indicated by altered procollagen-proline dioxygenase, protein disulfide isomerase, nucleoside diphosphate kinases, electron transfer flavoprotein and glutathione S-transferase. This work confirmed that an integration of proteomics and metabolomics could provide an insightful view into the effects of pathogens to the marine mussel M. galloprovincialis.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.