BackgroundSweet taste receptor is expressed not only in taste buds but also in nongustatory organs such as enteroendocrine cells and pancreatic beta-cells, and may play more extensive physiological roles in energy metabolism. Here we examined the expression and function of the sweet taste receptor in 3T3-L1 cells.Methodology/Principal FindingsIn undifferentiated preadipocytes, both T1R2 and T1R3 were expressed very weakly, whereas the expression of T1R3 but not T1R2 was markedly up-regulated upon induction of differentiation (by 83.0 and 3.8-fold, respectively at Day 6). The α subunits of Gs (Gαs) and G14 (Gα14) but not gustducin were expressed throughout the differentiation process. The addition of sucralose or saccharin during the first 48 hours of differentiation considerably reduced the expression of peroxisome proliferator activated receptor γ (PPARγ and CCAAT/enhancer-binding protein α (C/EBPα at Day 2, the expression of aP2 at Day 4 and triglyceride accumulation at Day 6. These anti-adipogenic effects were attenuated by short hairpin RNA-mediated gene-silencing of T1R3. In addition, overexpression of the dominant-negative mutant of Gαs but not YM-254890, an inhibitor of Gα14, impeded the effects of sweeteners, suggesting a possible coupling of Gs with the putative sweet taste-sensing receptor. In agreement, sucralose and saccharin increased the cyclic AMP concentration in differentiating 3T3-L1 cells and also in HEK293 cells heterologously expressing T1R3. Furthermore, the anti-adipogenic effects of sweeteners were mimicked by Gs activation with cholera toxin but not by adenylate cyclase activation with forskolin, whereas small interfering RNA-mediated knockdown of Gαs had the opposite effects.Conclusions3T3-L1 cells express a functional sweet taste-sensing receptor presumably as a T1R3 homomer, which mediates the anti-adipogenic signal by a Gs-dependent but cAMP-independent mechanism.

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Background: Insulin down-regulates GLUT4 by accelerating degradation in lysosomes. Results: Insulin through H 2 O 2 production dissociates retromer from LDM membrane in a protein kinase CK2-dependent manner. Conclusion: Insulin switches GLUT4 traffic route toward lysosomes via retromer inhibition. Significance: This revealed a unique oxidative stress-mediated insulin signal cascade that regulates the fate of GLUT4.

ACB may achieve faster mobilization ability recovery for patients after TKA without a reduction in analgesia when compared with FNB in the early postoperative period. However, due to the variations in the included studies, additional studies are needed to validate these conclusions.

The purpose of this study is to evaluate the survivorship and risk factors for radiographic progression and conversion to total hip arthroplasty (THA) after porous tantalum implant surgery in the treatment of osteonecrosis of the femoral head (ONFH). The study comprised 90 ONFH patients (104 consecutive hips) who were treated with a porous tantalum implant combined with bone grafting between June 2008 and December 2013. The patients were 19–61 years of age (mean age, 38 years). The mean follow-up was 42 months. The outcome measures included Harris hip score (HHS), radiographic outcome measures, and survivorship analysis with conversion to THA as the endpoint. The mean postoperative HHS was significantly lower than the mean preoperative HHS (P < 0.001). The Cox proportional hazards model showed that age and Association Research Circulation Osseous (ARCO) stage were independent risk factors for conversion to THA, while age, China-Japan Friendship Hospital (CJFH) type, and ARCO stage were independent risk factors for radiological progression. Ultimately, only 52.9% hips survived. Porous tantalum implant surgery combined with bone grafting is not a viable option for treating ONFH, especially in patients >35 years of age with preoperative ARCO stage III and CJFH type L3.

Perfusion decellularization with detergents is effective to maintain the architecture and proteins of extracellular matrix (ECM) for use in the field of lung tissue engineering (LTE). However, it is unclear which detergent is ideal to produce an acellular lung scaffold. In this study, we obtained two decellularized rat lung scaffolds using a novel detergent sodium lauryl ether sulfate (SLES) and a conventional detergent sodium dodecyl sulfate (SDS). Both decellularized lung scaffolds were assessed by histology, immunohistochemistry, scanning electron microscopy, DNA quantification, sulfated glycosaminoglycans (GAGs) quantification and western blot. Subsequently, the scaffolds were implanted subcutaneously in rats for 6 weeks and were evaluated via hematoxylin and eosin staining and Masson staining. Results indicated that SLES was effective to remove cells; moreover, lungs decellularized with SLES showed better preservation of sulfated GAGs, lung architecture, and ECM proteins than SDS. After 6 weeks, SLES scaffolds demonstrated a significantly greater potential for cell infiltration and blood vessel formation compared with SDS scaffolds. Taken together, we conclude that SLES is a promising detergent to produce an acellular scaffold using LTE for eventual transplantation.

The increasing aging of the world population is accompanied by a rise in the incidence of knee osteoarthritis (KOA). There has been a growing interest in shockwave treatment for orthopedic diseases, including KOA. In previous trials, extracorporeal shockwave therapy (ESWT) was compared to physical therapy or placebo in the treatment of KOA. However, the efficacy and safety of ESWT for KOA remains disputed. The present meta-analysis assessed the effects of ESWT in KOA. The PubMed, Medline, Embase, Web of Science, Research Gate and the Cochrane Library were searched to identify comparative studies involving ESWT for patients with KOA. The outcome indicators included the visual analog scale (VAS) score, range of motion (ROM), the Lequesne index (LI) and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC). In the comparison of the ESWT vs. placebo groups, the primary outcomes included the VAS score and ROM, while the LI was the secondary outcome. In the comparison of the ESWT vs. physical therapy groups, the primary outcomes included the pain score and ROM, while the secondary outcome was the WOMAC index. Relevant data were analyzed using RevMan v5.3. The ESWT group had a lower VAS core, larger ROM and a better LI than the placebo group after 1 month of therapy (P<0.05). Furthermore, at 1 month post-therapy, the ESWT group had a lower VAS score, larger ROM and a better WOMAC than the physical therapy group (P<0.05). The outcomes regarding pain, ROM, LI and WOMAC were significantly different between the two different groups (P<0.05). The present meta-analysis suggested that ESWT may achieve a better therapeutic effect for patients with KOA as compared to physical therapy. However, high-quality trials with large sample sizes are essential to substantiate these results.

The pro-inflammatory activation of pulmonary microvascular endothelial cells resulting in continuous expression of cellular adhesion molecules, and subsequently recruiting primed neutrophils to form a firm neutrophils-endothelium (PMN-EC) adhesion, has been examined and found to play a vital role in acute lung injury (ALI). RNA interference (RNAi) is a cellular process through harnessing a natural pathway silencing target gene based on recognition and subsequent degradation of specific mRNA sequences. It opens a promising approach for precision medicine. However, this application was hampered by many obstacles, such as immunogenicity, instability, toxicity problems, and difficulty in across the biological membrane. In this study, we reprogrammed urine exfoliated renal epithelial cells into human induced pluripotent stem cells (huiPSCs) and purified the exosomes (Exo) from huiPSCs as RNAi delivery system. Through choosing the episomal system to deliver transcription factors, we obtained a non-integrating huiPSCs. Experiments in both vitro and vivo demonstrated that these huiPSCs possess the pluripotent properties. The exosomes of huiPSCs isolated by differential centrifugation were visualized by transmission electron microscopy (TEM) showing a typical exosomal appearance with an average diameter of 122 nm. Immunoblotting confirmed the presence of the typical exosomal markers, including CD63, TSG 101, and Alix. Co-cultured PKH26-labeled exosomes with human primary pulmonary microvascular endothelial cells (HMVECs) confirmed that they could be internalized by recipient cells at a time-dependent manner. Then, electroporation was used to introduce siRNA against intercellular adhesion molecule-1 (ICAM-1) into exosomes to form an Exo/siRNA compound. The Exo/siRNA compound efficiently delivered the target siRNA into HMVECs causing selective gene silencing, inhibiting the ICAM-1 protein expression, and PMN-EC adhesion induced by lipopolysaccharide (LPS). These data suggest that huiPSCs exosomes could be used as a natural gene delivery vector to transport therapeutic siRNAs for alleviating inflammatory responses in recipient cells.