Glycation Improved the Interfacial Adsorption and Emulsifying Performance of ß-Conglycinin to Stabilize the High Internal Phase Emulsions.

This study investigated the interfacial adsorption and emulsifying performance of glycated ß-conglycinin (7S) with D-galactose (Gal) at various times. Results indicated that glycation increased the particle sizes and zeta potentials of glycated 7S by inducing subunit dissociation. Glycation destroyed the tertiary structures and transformed secondary structures from an ordered one to a disordered one, leading to the more flexible structures of glycated 7S compared with untreated 7S. All these results affected the structural unfolding and rearrangement of glycated 7S at the oil/water interface. Therefore, glycated 7S improved interfacial adsorption and formed an interfacial viscoelasticity layer, increasing emulsifying performance to stabilize high internal phase emulsions (HIPE) with self-supportive structures. Furthermore, the solid gel-like network of HIPE stabilized by glycated 7S led to emulsification stability. This result provided new ideas to improve the functional properties of plant proteins by changing the interfacial structure.

MiR-362 suppresses cervical cancer progression via directly targeting BAP31 and activating TGFß/Smad pathway.

BAP31 (B-cell receptor-associated protein 31) is an important regulator of intracellular signal transduction and highly expressed in several cancer tissues or testicular tissues. Our previous study had revealed that elevated BAP31 plays a crucial role in the progress and metastasis of cervical cancer. Even so, the precise mechanism of abnormal BAP31 elevation in cervical cancer has not been fully elucidated. We revealed that the expression of BAP31 was mainly regulated by microRNA-362 (miR-362), which was markedly downregulated in cervical cancer tissues and negatively correlated with clinical tumor staging. Overexpression of miR-362 inhibited cervical cancer cell proliferation and increased the proportion of apoptotic cells. Furthermore, miR-362 reduced the tumor sizes and prolonged mice survival time in xenograft nude mice model. Finally, we demonstrated that the BAP31/SPTBN1 complex regulated tumor progression through the Smad 2/3 pathway under the control of miR-362. Collectively, our findings demonstrated that miR-362 could work as an anti-oncomiR that inhibits proliferation and promotes apoptosis in cervical cancer cells via BAP31 and TGFß/Smad pathway. Overexpression of miR-362 might be a potential therapeutic strategy for cervical cancer.

MicroRNA-362-3p Inhibits Migration and Invasion via Targeting BCAP31 in Cervical Cancer.

Cervical cancer (CC) is the most common malignant tumor in gynecology, and metastasis is an important cause of patient death. MiRNAs (microRNAs) have been found to play key roles in cervical cancer metastasis, but the effect of miR-362-3p in CC is unclear. This study aimed to investigate the role of miR-362-3p in cervical cancer migration and invasion. We compared the expression levels of miR-362-3p in cervical cancer tissues and adjacent normal cervical tissues. In CC tissues, miR-362-3p expression was significantly down-regulated, which is related to the cancer stage and patient survival. MiR-362-3p can effectively inhibit the migration and invasion of cervical cancer cells. The dual-luciferase reporter assay results showed that BCAP31 (B cell receptor associated protein 31) is a direct target protein of miR-362-3p. The results of the immunohistochemical examination of clinical tissue samples showed that BCAP31 was abnormally highly expressed in cervical cancer, which was positively correlated with the clinical stage. BCAP31 knockdown exerted similar effects as miR-362-3p overexpression. Further GSEA analysis showed that BCAP31 may participate in multiple biological processes, such as protein transport, metabolism, and organelle organization. Our results suggest that miR-362-3p inhibits migration and invasion via directly targeting BCAP31 in cervical cancer, and restoring miR-362-3p levels may be a new treatment strategy for cervical cancer in the future.

BCAP31, a cancer/testis antigen-like protein, can act as a probe for non-small-cell lung cancer metastasis.

Non-small-cell lung cancer (NSCLC) represents most of lung cancers, is often diagnosed at an advanced metastatic stage. Therefore, exploring the mechanisms underlying metastasis is key to understanding the development of NSCLC. The expression of B cell receptor-associated protein 31 (BCAP31), calreticulin, glucose-regulated protein 78, and glucose-regulated protein 94 were analyzed using immunohistochemical staining of 360 NSCLC patients. It resulted that the high-level expression of the four proteins, but particularly BCAP31, predicted inferior overall survival. What's more, BCAP31 was closely associated with histological grade and p53 status, which was verified by seven cohorts of NSCLC transcript microarray datasets. Then, three NSCLC cell lines were transfected to observe behavior changes BCAP31 caused, we found the fluctuation of BCAP31 significantly influenced the migration, invasion of NSCLC cells. To identify the pathway utilized by BCAP31, Gene Set Enrichment Analysis was firstly performed, showing Akt/m-TOR/p70S6K pathway was the significant one, which was verified by immunofluorescence, kinase phosphorylation and cellular behavioral observations. Finally, the data of label-free mass spectroscopy implied that BCAP31 plays a role in a fundamental biological process. This study provides the first demonstration of BCAP31 as a novel prognostic factor related to metastasis and suggests a new therapeutic strategy for NSCLC.

Investigating the biomechanical function of the plate-type external fixator in the treatment of tibial fractures: a biomechanical study.

BACKGROUND: The design of an external fixator with the optimal biomechanical function and the lowest profile has been highly pursued, as fracture healing is dependent on the stability and durability of fixation, and a low profile is more desired by patients. The plate-type external fixator, a novel prototype of an external tibial fixation device, is a low profile construct. However, its biomechanical properties remain unclear. The objective of this study was to investigate the stiffness and strength of the plate-type external fixator and the unilateral external fixator. We hypothesized that the plate-type external fixator could provide higher stiffness while retaining sufficient strength. METHODS: Fifty-four cadaver tibias underwent a standardized midshaft osteotomy to create a fracture gap model to simulate a comminuted diaphyseal fracture. All specimens were randomly divided into three groups of eighteen specimens each and stabilized with either a unilateral external fixator or two configurations of the plate-type external fixator. Six specimens of each configuration were tested to determine fixation stiffness in axial compression, four-point bending, and torsion, respectively. Afterwards, dynamic loading until failure was performed in each loading mode to determine the construct strength and failure mode. RESULTS: The plate-type external fixator provided higher stiffness and strength than the traditional unilateral external fixator. The highest biomechanics were observed for the classical plate-type external fixator, closely followed by the extended plate-type external fixator. CONCLUSIONS: The plate-type external fixator is stiffer and stronger than the traditional unilateral external fixator under axial compression, four-point bending and torsion loading conditions.

Transcriptome Based System Biology Exploration Reveals Homogeneous Tumorigenicity of Alimentary Tract Malignancy.

Malignancies of alimentary tract include esophageal carcinoma (ESCA), stomach adenocarcinoma (STAD), colon adenocarcinoma (COAD), and rectum adenocarcinoma (READ). Despite of their similarities in cancer development and progression, there are numerous researches concentrating on single tumor but relatively little on their common mechanisms. Our study explored the transcriptomic data of digestive tract cancers from The Cancer Genome Atlas database, yielding their common differentially expressed genes including 1,700 mRNAs, 29 miRNAs, and 362 long non-coding RNAs (lncRNAs). There were 12 mRNAs, 5 miRNAs, and 16 lncRNAs in the core competitive endogenous RNAs network by RNA-RNA interactions, highlighting the prognostic nodes of SERPINE1, hsa-mir-145, and SNHG1. In addition, the weighted gene co-expression network analysis (WGCNA) illustrated 20 gene modules associated with clinical traits. By taking intersections of modules related to the same trait, we got 67 common genes shared by ESCA and READ and screened 5 hub genes, including ADCY6, CXCL3, NPBWR1, TAS2R38, and PTGDR2. In conclusion, the present study found that SERPINE1/has-mir-145/SNHG1 axis acted as promising targets and the hub genes reasoned the similarity between ESCA and READ, which revealed the homogeneous tumorigenicity of digestive tract cancers at the transcriptome level and led to further comprehension and therapeutics for digestive tract cancers.

Hantavirus Gc induces long-term immune protection via LAMP-targeting DNA vaccine strategy.

Hemorrhagic fever with renal syndrome (HFRS) occurs widely throughout Eurasia. Unfortunately, there is no effective treatment, and prophylaxis remains the best option against the major pathogenic agent, hantaan virus (HTNV), which is an Old World hantavirus. However, the absence of cellular immune responses and immunological memory hampers acceptance of the current inactivated HFRS vaccine. Previous studies revealed that a lysosome-associated membrane protein 1 (LAMP1)-targeting strategy involving a DNA vaccine based on the HTNV glycoprotein Gn successfully conferred long-term immunity, and indicated that further research on Gc, another HTNV antigen, was warranted. Plasmids encoding Gc and lysosome-targeted Gc, designated pVAX-Gc and pVAX-LAMP/Gc, respectively, were constructed. Proteins of interest were identified by fluorescence microscopy following cell line transfection. Five groups of 20 female BALB/c mice were subjected to the following inoculations: inactivated HTNV vaccine, pVAX-LAMP/Gc, pVAX-Gc, and, as the negative controls, pVAX-LAMP or the blank vector pVAX1. Humoral and cellular immunity were assessed by enzyme-linked immunosorbent assays (ELISAs) and 15-mer peptide enzyme-linked immunospot (ELISpot) epitope mapping assays. Repeated immunization with pVAX-LAMP/Gc enhanced adaptive immune responses, as demonstrated by the specific and neutralizing antibody titers and increased IFN-γ production. The inactivated vaccine induced a comparable humoral reaction, but the negative controls only elicited insignificant responses. Using a mouse model of HTNV challenge, the in vivo protection conferred by the inactivated vaccine and Gc-based constructs (with/without LAMP recombination) was confirmed. Evidence of pan-epitope reactions highlighted the long-term cellular response to the LAMP-targeting strategy, and histological observations indicated the safety of the LAMP-targeting vaccines. The long-term protective immune responses induced by pVAX-LAMP/Gc may be due to the advantage afforded by lysosomal targeting after exogenous antigen processing initiation and major histocompatibility complex (MHC) class II antigen presentation trafficking. MHC II-restricted antigen recognition effectively primes HTNV-specific CD4+ T-cells, leading to the promotion of significant immune responses and immunological memory. An epitope-spreading phenomenon was observed, which mirrors the previous result from the Gn study, in which the dominant IFN-γ-responsive hot-spot epitopes were shared between HLA-II and H2d. Importantly, the pan-epitope reaction to Gc indicated that Gc should be with potential for use in further hantavirus DNA vaccine investigations.

Closed reduction and percutaneous annulated screw fixation in the treatment of comminuted proximal humeral fractures.

BACKGROUND: Displaced proximal humeral fractures remain a challenge to orthopedic surgeons. OBJECTIVES: The purpose of this study was to evaluate the functional and radiological outcomes of patients with comminuted proximal humeral fractures treated with closed reduction and percutaneous screw fixation (CRPF). MATERIAL AND METHODS: The authors retrospectively reviewed 38 cases of displaced proximal humeral fractures (2-, 3- or 4-part fractures according to the Neer classification) that were treated using the CRPF technique from May 2009 to April 2013. From this group 26 patients were followed up for a period ranging from 9 to 24 months (averaging 12.9 months) and evaluated for the functional and radiological outcomes by a series of standard questionnaires and measurements. RESULTS: The fractures in all 26 patients were healed within an average time of 14.6 weeks (ranging from 11 to 27 weeks), and the mean interval between the operation and fully functional activity was 18.6 weeks (ranging from 15 to 32 weeks). At the final follow-up visit, no patient showed shoulder instability; the mean range of abduction motion was 146.5° (ranging from 72° to 180°). For all patients, no statistically significant difference in the functional outcomes was observed between their 6-month and final follow-up visits; or in the radiological findings between their immediate post-operative and final follow-up examinations. CONCLUSIONS: The CRPF technique is a safe and effective therapeutic option for comminuted proximal humeral fractures. Good stability is obtained and aggressive impairment of the soft tissue and periosteum around the fracture is avoided, which allows for an early painless range of motion. The technique promotes bone healing, prevents ischemic osteonecrosis of the head of the humerus and leads to few complications.

Orthopedic injury in electric bicycle-related collisions.

OBJECTIVE: Although electric bicycle-related injuries have become the most common reason for hospitalization due to a road crash in China, no study has comprehensively investigated electric bicycle collisions and their impact on orthopedic injuries; such a study may provide evidence to support a new road safety policy. METHODS: A retrospective review of orthopedic injuries from electric bicycle collisions was performed in an urban trauma center. We collected variables including age, gender, location of fracture, presence of open or closed fractures, concomitant vascular, and neurologic injuries. RESULTS: A total of 2,044 cases were involved in electric bicycle collisions. The orthopedic injury victims were predominantly male and middle aged. The most common orthopedic injury was a femur fracture. Open fractures frequently involved the forearm and tibia/fibula. Male patients were more likely to suffer from multiple fractures and associated injuries than female patients. Fewer patients age 60 years old or older wore helmets at the time of the accident compared to those in other age groups. CONCLUSIONS: Orthopedic injuries from electric bicycle-related accidents cause patients substantial suffering that could lead to serious social consequences. Helmet use and protective clothing or similar safety gear, especially for electric bicycle users, should be required to provide greater protection.

Tanshinone IIA attenuates hypoxic pulmonary hypertension via modulating KV currents.

The voltage-gated K(+) (KV) channels play an essential role in the etiology of chronic hypoxic pulmonary hypertension (CH-PH).Tanshinone IIA (TIIA), a major active component of Salvia miltiorrhiza Bunge (S. miltiorrhiza), has many biological protective effects. In the present study, we investigated whether KV channels were responsible for the protective effect of TIIA on CH-PH. In acute hypoxia experiments, the IKV currents of pulmonary artery smooth muscle cells (PASMCs) isolated from healthy rats were determined in the absence or presence of TIIA (5 µg/ml or 25 µg/ml) or 4-AP (1 mM). In chronic hypoxia experiments, rats were challenged by intermittent hypoxia or sustained hypoxia exposure for 4 weeks with or without TIIA (10 mg/kg) treatment. Subsequently, the hemodynamic data and the pathomorphological changes of pulmonary arteries were gathered. The expressions of KV2.1 and KV1.5 in pulmonary arteries were tested by Western blotting and RT-PCR, respectively. PASMCs were detached from intermittent hypoxia or sustained hypoxia exposure rats to evaluate the IKV currents. Results showed that TIIA markedly recovered acute hypoxia-induced the down-regulation of IKV currents in PASMCs. Moreover, TIIA significantly restrained chronic intermittent hypoxia or sustained hypoxia-induced pulmonary artery wall remodeling, accompanied with modulating the expressions of KV2.1 and KV1.5, and reversing the down-regulation of IKV currents. TIIA is thus an attractive potential therapy for CH-PH.

Neuroprotective effects of daphnetin against NMDA receptor-mediated excitotoxicity.

The accumulation of glutamate can excessively activate the N-methyl-d-aspartate (NMDA) receptors and cause excitotoxicity. Daphnetin (Dap), a coumarin derivative, is a protein kinase inhibitor that exhibits antioxidant and neuroprotective properties. However, little is known about the neuroprotective effects of Dap on glutamate-induced excitotoxicity. We evaluated the neuroprotective activities in the primary cultured cortical neurons against NMDA-induced excitotoxicity. Pretreatment with Dap significantly prevented NMDA-induced neuronal cell loss. Dap significantly inhibited the neuronal apoptosis by regulating balance of Bcl-2 and Bax expression. Furthermore, pretreatment of Dap reversed the up-regulation of NR2B-containing NMDA receptors and inhibited the intracellular Ca2+ overload induced by NMDA exposure. In addition, Dap prevented cerebral ischemic injury in mice induced via a 2 h middle cerebral artery occlusion and a 24 h reperfusion in vivo. The findings suggest that Dap prevents the excitotoxicity through inhibiting the NR2B-containing NMDA receptors and the subsequent calcium overload in cultured cortical neurons.

Fasudil reversed MCT-induced and chronic hypoxia-induced pulmonary hypertension by attenuating oxidative stress and inhibiting the expression of Trx1 and HIF-1α.

Antioxidant therapy attenuates pulmonary hypertension (PH). In the present study, we tested the antioxidant effects of fasudil against PH in rats. Monocrotaline (MCT)-induced and chronic hypoxia-induced PH models of rats were established, and the haemodynamic and pathomorphologic results of three different doses of fasudil (10 mg/kg, 30 mg/kg, and 75 mg/kg per day) were subsequently compared with those of bosentan (30 mg/kg per day). Additionally, the protein expressions of thioredoxin-1 (Trx1) and hypoxia inducible factor-1α (HIF-1α), the content of superoxide dismutase (SOD), and the levels of hydrogen peroxide (H2O2), malonyldialdehyde (MDA), and hydroxy radical (·OH) were investigated. Fasudil effectively reduced the right ventricular systolic pressure (RVSP) and alleviated right ventricle (RV) hypertrophy, as well as the histological changes in the pulmonary arterioles. Moreover, fasudil markedly lessened the expression of Trx1 and HIF-1α, up-regulated the concentration of SOD, and lowered the levels of H2O2, MDA, and ·OH. In conclusion, fasudil is a notably attractive potential therapy for PH.

mTOR signal transduction pathways contribute to TN-C FNIII A1 overexpression by mechanical stress in osteosarcoma cells.

Osteosarcoma is the most common primary malignant bone tumor with a very poor prognosis. Treating osteosarcoma remains a challenge due to its high transitivity. Tenascin-C, with large molecular weight variants including different combinations of its alternative spliced FNIII repeats, is specifically over expressed in tumor tissues. This study examined the expression of Tenascin-C FNIIIA1 in osteosarcoma tissues, and estimated the effect of mechanical stimulation on A1 expression in MG-63 cells. Through immunohistochemical analysis, we found that the A1 protein was expressed at a higher level in osteosarcoma tissues than in adjacent normal tissues. By cell migration assay, we observed that there was a significant correlation between A1 expression and MG-63 cell migra-tion. The relation is that Tenascin-C FNIIIA1 can promote MG-63 cell migration. According to our further study into the effect of mechanical stimulation on A1 expression in MG-63 cells, the mRNA and protein levels of A1 were significantly up-regulated under mechanical stress with the mTOR molecule proving indispensable. Meanwhile, 4E-BP1 and S6K1 (downstream molecule of mTOR) are necessary for A1 normal expression in MG-63 cells whether or not mechanical stress has been encountered. We found that Tenascin-C FNIIIA1 is over-expressed in osteosar-coma tissues and can promote MG-63 cell migration. Furthermore, mechanical stress can facilitate MG-63 cell migration though facilitating A1 overexpression with the necessary molecules (mTOR, 4E-BP1 and S6K1). In con-clusion, high expression of A1 may promote the meta-stasis of osteosarcoma by facilitating MG-63 cell migration. Tenascin-C FNIIIA1 could be used as an indicator in metastatic osteosarcoma patients.

Purified dendritic cell-tumor fusion hybrids supplemented with non-adherent dendritic cells fraction are superior activators of antitumor immunity.

BACKGROUND: Strong evidence supports the DC-tumor fusion hybrid vaccination strategy, but the best fusion product components to use remains controversial. Fusion products contain DC-tumor fusion hybrids, unfused DCs and unfused tumor cells. Various fractions have been used in previous studies, including purified hybrids, the adherent cell fraction or the whole fusion mixture. The extent to which the hybrids themselves or other components are responsible for antitumor immunity or which components should be used to maximize the antitumor immunity remains unknown. METHODS: Patient-derived breast tumor cells and DCs were electro-fused and purified. The antitumor immune responses induced by the purified hybrids and the other components were compared. RESULTS: Except for DC-tumor hybrids, the non-adherent cell fraction containing mainly unfused DCs also contributed a lot in antitumor immunity. Purified hybrids supplemented with the non-adherent cell population elicited the most powerful antitumor immune response. After irradiation and electro-fusion, tumor cells underwent necrosis, and the unfused DCs phagocytosed the necrotic tumor cells or tumor debris, which resulted in significant DC maturation. This may be the immunogenicity mechanism of the non-adherent unfused DCs fraction. CONCLUSIONS: The non-adherent cell fraction (containing mainly unfused DCs) from total DC/tumor fusion products had enhanced immunogenicity that resulted from apoptotic/necrotic tumor cell phagocytosis and increased DC maturation. Purified fusion hybrids supplemented with the non-adherent cell population enhanced the antitumor immune responses, avoiding unnecessary use of the tumor cell fraction, which has many drawbacks. Purified hybrids supplemented with the non-adherent cell fraction may represent a better approach to the DC-tumor fusion hybrid vaccination strategy.

Tumor immunotherapy based on tumor-derived heat shock proteins (Review).

Heat shock proteins (HSPs), the most important type of molecular chaperone, are expressed in all eukaryotic cells and have multiple functions, including the folding and unfolding of other proteins and peptides, the transport of proteins and peptides and the support of antigen presentation processes. Due to these important properties, the use of HSPs has been explored as a promising tumor immunotherapy strategy. It has been previously demonstrated that HSP peptide complex (HSP.PC) derived from tumors is the immunogenic entity that elicits powerful antitumor immune responses. Previous animal studies and phase III clinical trials have demonstrated the efficacy, safety and feasibility of HSP-based tumor vaccines. However, the limitations are also apparent and specific alternatives have been developed. The present review focused on the history of HSP-based immunotherapy, the mechanism of its immunogenicity and the previous efforts to promote the efficacy. The current review may be useful for antitumor studies based on the tumor-derived HSPs.

Mechanical strain and growth factors regulate expression of tenascin-C by OS cells additively.

Osteosarcoma (OS) is a kind of malignancy wherein the tumor cells form malignant bone-like or bone tissue. Tenascin-C (TN-C), an important extracellular matrix (ECM) protein, plays an indispensable role in tumor development. However, its regulatory factors, expression, and function in OS pathological process have not been studied extensively. Expression of TN-C is induced by growth factors as well as mechanical strain in fibroblast. So we asked whether mechanical stain and growth factors could induce TN-C expression in OS as well as which pathways were involved in those processes. We found that when mechanical strain was applied to OS cells cultured on silicone membrane, TN-C mRNA and protein levels were increased 10-fold within 8 h compared to the resting control. Likewise, when epidermal growth factors (EGFs) and insulin-like growth factor (IGF-1) were added to cells, TN-C mRNA levels increased six fold and eightfold, respectively, within 24h compared to the control. Growth factors (EGF and IGF-1) and mechanical strain had additive effects on the induction of TN-C mRNA expression in OS. Both ROCK-I/II inhibitor and MEK-1 inhibitor inhibited TN-C induction by EGF or IGF-1, while only ROCK-I/II inhibitor had a strong subdued effect on TN-C induction by mechanical strain. Taken together, our findings suggest that growth factors and mechanical strain can induce TN-C in OS through different pathways additively.

Tumoricidal effects of a selenium (Se)-polysaccharide from Ziyang green tea on human osteosarcoma U-2 OS cells.

Selenium(Se)-enriched green tea consumption in human diets is well-known to reduce the risk of a variety of diseases. Here, we isolated a Se-polysaccharide (Se-ZYTP) from Se-enriched Ziyang green tea and investigated its antitumor activity on human osteosarcoma U-2 OS cells in vitro and in vivo. Se-ZYTP contained 94.5% of carbohydrate and 2.1% of uronic acid, as well as 2.14 µg/g Se, revealing that Se-ZYTP was an acidic Se-conjugated polysaccharide. Monosaccharide composition analysis indicated that Se-ZYTP consisted of mannose, rhamnose and fucose in molar ratios of 2.4:1.5:1.2:0.2:0.1:0.3:0.3. In vitro, both MTT and LTH assays proved that Se-ZYTP (25, 50, 100 and 200 µg/ml) could significantly inhibit the proliferation of human osteosarcoma U-2 OS cells in a concentration-dependent fashion (P<0.05 or P<0.01). In U-2 OS cancer xenograft model in BALB/c athymic mice, Se-ZYTP oral administration at three doses of 100, 200 and 400mg/kg body weight (B.W.) daily for 28 days resulted in obvious tumor regression as compared to model control (P<0.05 or P<0.01). In addition, body weights of mice in control or Se-ZYTP treated groups did not differ significantly and no mice died during experiment, suggesting the safety of Se-ZYTP. Therefore, we postulate that Se-ZYTP might have cancer-preventive and cancer-therapeutic benefit for human osteosarcoma.

Conditioned medium of Wnt/ß-catenin signaling-activated olfactory ensheathing cells promotes synaptogenesis and neurite growth in vitro.

Olfactory ensheathing cells (OECs), the major glia cells in the olfactory system, have been extensively studied because of their ability to promote axonal growth and regeneration. Whether it could facilitate synaptogenesis is an important, but remains as yet an unanswered question. We have identified a subgroup of Wnt signaling-activated OECs, spatiotemporal distribution of which in the olfactory bulb suggests a role for these cells in both axonal growth and synaptogenesis. In the present study, we explored this possibility in vitro. OECs were primarily cultured, in which Wnt signaling was activated by overexpressing ß-catenin, and inhibited by dominant negative TCF4. Neurite growth and synaptogenesis were assessed by co-culturing neurons with conditioned medium from control OECs (cOECs CM), Wnt/ß-catenin signaling-activated OECs (wOECs CM), or Wnt signaling-inhibited OECs (wiOECs). The results showed that although cOECs CM enhances axonal growth, wOECs CM exhibited a stronger axonal growth-promoting effect, than cOECs CM. More importantly, wOECs CM stimulates synatpogenesis, demonstrated by the expression of Synaptophysin and whole-cell patch clamp recording. In contrast, both cOECs CM and wiOECs CM do not affect synaptogenesis. Our data, for the first time, demonstrated that, in comparison with regularly cultured OECs, wOECs CM are more effective in enhancing axonal growth, and can promote synaptogenesis, probably by secreting factors. These results suggest a potential application of wOECs for treating spinal cord injury.

miR-34a inhibits the metastasis of osteosarcoma cells by repressing the expression of CD44.

Osteosarcoma is the most common type of malignant bone tumor in children and adolescents and approximately 30% of patients develop lung metastasis, which is the leading cause of mortality. In this study, we investigated the role of miR-34a in the invasion and metastasis of osteosarcoma cells by examining its expression level and functional pattern in these cells. miR-34a mimics were transfected into the highly metastatic subline, F5M2, and into the F4 subline with low metastatic potential of the paired human osteosarcoma cell line, SOSP­9607. Cell viability patterns, cell migration and alterations in gene expression levels were assessed by real-time PCR, and changes in protein levels were assessed by immunocytochemistry and western blot analysis. The ectopic overexpression of miR-34a significantly inhibited the migration and invasive ability of osteosarcoma cells by repressing the expression of CD44. These data suggest that miR-34a plays a tumor suppressor role in the metastasis of osteosarcoma cells by repressing the expression of CD44. Of note, studies have also suggested that the CD44 protein correlates with the metastatic potential of several malignant tumors. Therefore, it can be concluded that through the inhibition of CD44 expression levels, miR-34a plays a significant role in the migration and invasion of osteosarcoma cells.

Neuroprotective effects of formononetin against NMDA-induced apoptosis in cortical neurons.

Formononetin (FMNT) is an isoflavone found in many herbs including Trifolium pratense L., Spatholobus suberectus Dunn., and Astragalus mongholicus Bunge. The purpose of this study is to investigate pharmacological properties of FMNT on neurotoxicity induced by N-methyl-D-asparate (NMDA) in primary-cultured cortical neurons. The cell viability was significantly decreased after exposure to NMDA (200 µM) for 40 min. Pretreatment of FMNT (10 µM) for 12 h significantly attenuated the cell loss induced by NMDA exposure. Flow cytometry analysis revealed that treatment of FMNT attenuated the number of apoptotic cells, especially the early phase apoptotic cells, induced by NMDA exposure. Western blot analysis showed that FMNT regulated the expression of apoptosis-related proteins by increasing the levels of Bcl-2 and pro-caspase-3 and decreasing the levels of Bax and caspase-3. These findings demonstrate that FMNT is capable of protecting neurons from NMDA-evoked excitotoxic injury and has a potential perspective to the clinical treatment for neurodegenerative disorders in central nervous system.