Quantifying Fluid Shear Stress in a Rocking Culture Dish

Fluid shear stress (FSS) is an important stimulus for cell functions. Compared with the well established parallel-plate and cone-and-plate systems, a rocking “see-saw” system offers some advantages such as easy operation, low cost, and high throughput. However, the FSS spatiotemporal pattern in the system has not been quantified. In the present study, we developed a lubrication-based model to analyze the FSS distributions in a rocking rectangular culture dish. We identified an important parameter (the critical flip angle) that dictates the overall FSS behaviors and suggested the right conditions to achieving temporally oscillating and spatially relatively uniform FSS. If the maximal rocking angle is kept smaller than the critical flip angle, which is defined as the angle when the fluid free surface intersects the outer edge of the dish bottom, the dish bottom remains covered with a thin layer of culture medium. The spatial variations of the peak FSS within the central 84% and 50% dish bottom are limited to 41% and 17%, respectively. The magnitude of FSS was found to be proportional to the fluid viscosity and the maximal rocking angle, and inversely proportional to the square of the fluid depth-to-length ratio and the rocking period. For a commercial rectangular dish (length of 37.6 mm) filled with ∼2 mL culture medium, the FSS at the center of the dish bottom is expected to be on the order of 0.9 dyn/cm2 when the dish is rocked +5° at 1 cycle/s. Our analysis suggests that a rocking “see-saw” system, if controlled well, can be used as an alternative method to provide low-magnitude, dynamic FSS to cultured cells

Financing Mode of American Small and Medium-sized Enterprises and the Enlightenment to Our Country

Because of the characteristic of small and medium-sized enterprises such as small scale of assets, low transparency message and heavy uncertainty of management, the financing mode of small and medium-sized enterprises of various countries is the same basically. This paper reflect the characteristic of the financing mode of small and medium-sized enterprises of our country indirectly through studying the financing mode and characteristics of American small and medium-sized enterprises, expecting to enlighten alleviating the difficulty of financing of small and medium-sized enterprises of our country . Key words: Financing mode, Characteristic, Enlightenment Résumé : La modèle de fusion des petites et moyennes entreprises dans tous les pays existe des points communs , en raison de ses quelques caractéristiques suivantes : les capitaux de petite envergure , la transparence insuffisante des informations et l’instabilité de gestion . A travers la recherche sur la modèle de fusion des petites et moyennes entreprises aux états-unis , ce texte reflète de façon indirecte les traits de modèle de fusion des petites et moyennes entreprises en Chine dans l’espoir de donner l’inspiration à l’atténuation des difficultés de la fusion des petites et moyennes entreprises de notre pays . Mots-clés: modèle de diffusion, traits , inspiration 摘要：由於中小企業具有資產規模小，資訊透明度低，經營不確定性大等特徵，各國中小企業融資模式存在一定的共同之處。本文通過研究美國中小企業融資模式及其特徵，間接反映了我國中小企業融資模式的特點，期望對緩解我國中小企業融資困難有所啟示。 關鍵詞：融資模式；特徵；啟

Effect of Low-magnitude, High-frequency Vibration on Osteocytes in the Regulation of Osteoclasts

Osteocytes are well evidenced to be the major mechanosensor in bone, responsible for sending signals to the effector cells (osteoblasts and osteoclasts) that carry out bone formation and resorption. Consistent with this hypothesis, it has been shown that osteocytes release various soluble factors (e.g. transforming growth factor-β, nitric oxide, and prostaglandins) that influence osteoblastic and osteoclastic activities when subjected to a variety of mechanical stimuli, including fluid flow, hydrostatic pressure, and mechanical stretching. Recently, low-magnitude, high-frequency (LMHF) vibration (e.g., acceleration less than \u3c 1 × g, where g = 9.81 m/s2, at 20–90 Hz) has gained much interest as studies have shown that such mechanical stimulation can positively influence skeletal homeostasis in animals and humans. Although the anabolic and anti-resorptive potential of LMHF vibration is becoming apparent, the signaling pathways that mediate bone adaptation to LMHF vibration are unknown. We hypothesize that osteocytes are the mechanosensor responsible for detecting the vibration stimulation and producing soluble factors that modulate the activity of effector cells. Hence, we applied low-magnitude (0.3 × g) vibrations to osteocyte-like MLO-Y4 cells at various frequencies (30, 60, 90 Hz) for 1 h. We found that osteocytes were sensitive to this vibration stimulus at the transcriptional level: COX-2 maximally increased by 344% at 90 Hz, while RANKL decreased most significantly (−55%, p \u3c 0.01) at 60 Hz. Conditioned medium collected from the vibrated MLO-Y4 cells attenuated the formation of large osteoclasts (≥ 10 nuclei) by 36% (p \u3c 0.05) and the amount of osteoclastic resorption by 20% (p = 0.07). The amount of soluble RANKL (sRANKL) in the conditioned medium was found to be 53% lower in the vibrated group (p \u3c 0.01), while PGE2 release was also significantly decreased (−61%, p \u3c 0.01). We conclude that osteocytes are able to sense LMHF vibration and respond by producing soluble factors that inhibit osteoclast formation

Local stimulation of osteocytes using a magnetically actuated oscillating beam.

Mechanical loading on bone tissue is an important physiological stimulus that plays a key role in bone growth, fracture repair, and treatment of bone diseases. Osteocytes (bone cells embedded in bone matrix) are well accepted as the sensor cells to mechanical loading and play a critical role in regulating the bone structure in response to mechanical loading. To understand the response of osteocytes to differential mechanical stimulation in physiologically relevant arrangements, there is a need for a platform which can locally stimulate bone cells with different levels of fluid shear stress. In this study, we developed a device aiming to achieve non-contact local mechanical stimulation of osteocytes with a magnetically actuated beam that generates the fluid shear stresses encountered in vivo. The stimulating beam was made from a composite of magnetic powder and polymer, where a magnetic field was used to precisely oscillate the beam in the horizontal plane. The beam is placed above a cell-seeded surface with an estimated gap height of 5 μm. Finite element simulations were performed to quantify the shear stress values and to generate a shear stress map in the region of interest. Osteocytes were seeded on the device and were stimulated while their intracellular calcium responses were quantified and correlated with their position and local shear stress value. We observed that cells closer to the oscillating beam respond earlier compared to cells further away from the local shear stress gradient generated by the oscillating beam. We have demonstrated the capability of our device to mimic the propagation of calcium signalling to osteocytes outside of the stimulatory region. This device will allow for future studies of osteocyte network signalling with a physiologically accurate localized shear stress gradient

Vibration Therapy for Cancer-Related Bone Diseases

Patients undergoing cancer treatments and/or suffering from metastatic bone lesions experience various skeletal-related events (SREs), substantially reducing functional independence and quality of life. Therefore, researchers are working towards developing new interventions by harnessing the bone’s innate anabolic response to mechanical stimulations. Whole body vibration (WBV) has recently gained interest due to its nature of being safe, effective, and easy to perform. In this review, we will summarize the most cutting-edge vibration studies of cancer models and bone-cancer cell interactions. We will also discuss various parameters, including age, vibration settings, and differences between bone sites, which may affect vibration efficacy. Studies have shown that WBV improves bone mineral density (BMD) and bone volume in patients and mice with cancer. WBV also reduces tumor burden and normalizes bone vasculature in mice. At the cellular level, vibration promotes interactions between bone cells and cancer cells, which reduce osteoclastogenesis and inhibit cancer metastatic potential. Hence, WBV could potentially serve as a new intervention or adjuvant treatment to attenuate cancer progression while preserving bone health

A systematic review and meta-analysis of the benefits of a gluten-free diet and/or casein-free diet for children with autism spectrum disorder

ContextIt has been suggested that a gluten-free and casein-free (GFCF) diet may alleviate the symptoms of autism spectrum disorder (ASD) and facilitate neurodevelopment of children with ASD. Studies to date have been inconclusive.ObjectiveThis study aimed to evaluate (through quantitative meta-analysis) the efficacy and safety of a GFCF diet for children with ASD. To our knowledge, this is the first time such an analysis has been carried out.Data sourcesEight electronic databases were searched, from the establishment of each database up to March 27, 2020: PubMed, Web of Science, Embase (Ovid), PsycINFO (Ovid), Cochrane Library, CNKI, Wanfang, and VIP databases.Data extractionTwo authors independently performed the data extraction and risk-of-bias assessment.Data analysisA quantitative meta-analysis was performed with standard procedures by using Stata SE 15 software. Within the total of 8 studies, with 297 participants, 5 studies reported significant reductions in stereotypical behaviors [standard mean difference (SMD) = -0.41, 95% confidence interval (CI): -0.68 to -0.15], and 3 studies reported improvements in cognition (SMD = -0.46, 95% CI: -0.91 to -0.01) following GFCF dietary intervention . No statistically significant changes were observed in other symptomatic categories (all P > 0.05).ConclusionThe current meta-analysis showed that a GFCF diet can reduce stereotypical behaviors and improve the cognition of children with ASD. Though most of the included studies were single-blind, the benefits of a GFCF diet that have been indicated are promising. Additional studies on a larger scale are warranted.Systematic review registrationPROSPERO registration no. CRD42020177619

Yoda1 Enhanced Low-Magnitude High-Frequency Vibration on Osteocytes in Regulation of MDA-MB-231 Breast Cancer Cell Migration

Low-magnitude (≤1 g) high-frequency (≥30 Hz) (LMHF) vibration has been shown to enhance bone mineral density. However, its regulation in breast cancer bone metastasis remains controversial for breast cancer patients and elder populations. Yoda1, an activator of the mechanosensitive Piezo1 channel, could potentially intensify the effect of LMHF vibration by enhancing the mechanoresponse of osteocytes, the major mechanosensory bone cells with high expression of Piezo1. In this study, we treated osteocytes with mono- (Yoda1 only or vibration only) or combined treatment (Yoda1 and LMHF vibration) and examined the further regulation of osteoclasts and breast cancer cells through the conditioned medium. Moreover, we studied the effects of combined treatment on breast cancer cells in regulation of osteocytes. Combined treatment on osteocytes showed beneficial effects, including increasing the nuclear translocation of Yes-associated protein (YAP) in osteocytes (488.0%, p < 0.0001), suppressing osteoclastogenesis (34.3%, p = 0.004), and further reducing migration of MDA-MB-231 (15.1%, p = 0.02) but not Py8119 breast cancer cells (4.2%, p = 0.66). Finally, MDA-MB-231 breast cancer cells subjected to the combined treatment decreased the percentage of apoptotic osteocytes (34.5%, p = 0.04) but did not affect the intracellular calcium influx. This study showed the potential of stimulating Piezo1 in enhancing the mechanoresponse of osteocytes to LMHF vibration and further suppressing breast cancer migration via osteoclasts