Effects of miR-29c on proliferation and adipogenic differentiation of porcine bone marrow mesenchymal stromal cells.

microRNAs (miRNAs), a subclass of noncoding short RNAs, direct cells fate decisions that are important for cell proliferation and cell lineage decisions. Adipogenic differentiation contributes greatly to the development of white adipose tissue, involving of highly organized regulation by miRNAs. In the present study, we screened and identified 78 differently expressed miRNAs of porcine BMSCs during adipogenic differentiation. Of which, the role of miR-29c in regulating the proliferation and adipogenic differentiation was proved and detailed. Specifically, over-expression miR-29c inhibits the proliferation and adipogenic differentiation of BMSCs, which were reversed upon miR-29c inhibitor. Interference of IGF1 inhibits the proliferation and adipogenic differentiation of BMSCs. Mechanistically, miR-29c regulates the proliferation and adipogenic differentiation of BMSCs by targeting IGF1 and further regulating the MAPK pathway and the PI3K-AKT-mTOR pathway, respectively. In conclusion, we highlight the important role of miR-29c in regulating proliferation and adipogenic differentiation of BMSCs.

Kindlin-2 controls angiogenesis through modulating Notch1 signaling.

Kindlin-2 is critical for development and homeostasis of key organs, including skeleton, liver, islet, etc., yet its role in modulating angiogenesis is unknown. Here, we report that sufficient KINDLIN-2 is extremely important for NOTCH-mediated physiological angiogenesis. The expression of KINDLIN-2 in HUVECs is significantly modulated by angiogenic factors such as vascular endothelial growth factor A or tumor necrosis factor α. A strong co-localization of CD31 and Kindlin-2 in tissue sections is demonstrated by immunofluorescence staining. Endothelial-cell-specific Kindlin-2 deletion embryos die on E10.5 due to hemorrhage caused by the impaired physiological angiogenesis. Experiments in vitro show that vascular endothelial growth factor A-induced multiple functions of endothelial cells, including migration, matrix proteolysis, morphogenesis and sprouting, are all strengthened by KINDLIN-2 overexpression and severely impaired in the absence of KINDLIN-2. Mechanistically, we demonstrate that KINDLIN-2 inhibits the release of Notch intracellular domain through binding to and maintaining the integrity of NOTCH1. The impaired angiogenesis and avascular retinas caused by KINDLIN-2 deficiency can be rescued by DAPT, an inhibitor of γ-secretase which releases the intracellular domain from NOTCH1. Moreover, we demonstrate that high glucose stimulated hyperactive angiogenesis by increasing KINDLIN-2 expression could be prevented by KINDLIN-2 knockdown, indicating Kindlin-2 as a potential therapeutic target in treatment of diabetic retinopathy. Our study for the first time demonstrates the significance of Kindlin-2 in determining Notch-mediated angiogenesis during development and highlights Kindlin-2 as the potential therapeutic target in angiogenic diseases, such as diabetic retinopathy.

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC50 (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z' factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.

Anticancer fungal natural products: Mechanisms of action and biosynthesis.

Many fungal metabolites show promising anticancer properties both in vitro and in animal models, and some synthetic analogs of those metabolites have progressed into clinical trials. However, currently, there are still no fungi-derived agents approved as anticancer drugs. Two potential reasons could be envisioned: 1) lacking a clear understanding of their anticancer mechanism of action, 2) unable to supply enough materials to support the preclinical and clinic developments. In this review, we will summarize recent efforts on elucidating the anticancer mechanisms and biosynthetic pathways of several promising anticancer fungal natural products.

Management of early-stage gastro-esophageal cancers: expert perspectives from the Australasian Gastrointestinal Trials Group (AGITG) with invited international faculty.

Introduction: A multimodal approach in operable early-stage oesophago-gastric (OG) cancer has evolved in the last decade, leading to improvement in overall outcomes.Areas covered: A review of the published literature and conference abstracts was undertaken on the topic of optimal adjunctive chemotherapy or chemoradiotherapy in early-stage OG cancers. This review article focuses on the current evidence pertaining to neoadjuvant and perioperative strategies in curable OG cancers including the evolving landscape of immunotherapy and targeted drugs in this setting.Expert commentary: Adjunctive therapies in the form of preoperative chemo-radiotherapy (CRT) or chemotherapy and perioperative chemotherapy over surgery alone improve outcomes in patients with operable OG cancer. Although there are variations in practice around the world, a multi-disciplinary approach to patient care is of paramount importance. Immunotherapy and on treatment functional imaging are two examples of emerging strategies to improve the outcome for early-stage patients. A better understanding of the molecular biology of this disease may help overcome the problem of tumor heterogeneity and enable more rationally designed and targeted therapeutic interventions in the future.

Efficacy Evaluation of a non-contact automatic articulating paper dispenser in controlling articulating paper microbial contamination.

This study is to quantitatively evaluate the efficacy of using a non-contact automatic articulating paper dispenser for reducing microbial articulating paper contamination. Articulating papers in four-handed mode, non-four-handed mode, and via an automatic articulating paper dispenser were evaluated. An adenosine triphosphate bioluminescence assay was used to quantitatively measure the relative light unit (RLU) values of the rest unused articulating papers in the same package to detect contamination at 4 time points, and triplicate examinations were performed for all three methods. The RLUs were recorded, compared, and evaluated. For four-handed mode (n = 36), the RLUs at the four time points were 2.44, 32.89, 37.89, and 27.22, with a satisfactory rate of 94%. The RLUs for non-four-handed mode (n = 36) were 2.22, 286.44, 299.44, and 493.56, with a satisfactory rate of 36%. The RLUs for using the automatic dispenser (n = 36) were all 0 with a satisfactory rate of 100%. The satisfactory rates were significantly different among three methods. No significant differences were observed in the satisfactory rates for the four time points samples. Contact by gloved hands can cause severe biological contamination of articulating paper. However, by using standard four-handed mode or a non-contact automatic articulating paper dispenser, contamination can be controlled.

The mechanically activated p38/MMP-2 signaling pathway promotes bone marrow mesenchymal stem cell migration in rats.

OBJECTIVE: The aim of the present study was to investigate the effect of static strain on bone marrow mesenchymal stem cell (BMMSC) migration and whether the p38/matrix metalloproteinase-2 (MMP-2) axis plays a role in induction of BMMSC migration under mechanical strain. DESIGN: Both in vivo and in vitro investigations were performed. Twelve adult male Sprague-Dawley rats were randomly divided into 2 groups (n=6 per group). Rats in the experimental group underwent right mandibular distraction osteogenesis, whereas rats in the control group were subjected to osteotomy in the mandible without distraction. Immunohistochemistry and immunofluorescence were performed to evaluate phospho-p38 (p-p38) and Nestin expression. BMMSCs were isolated from rat mandibles. BMMSCs in the experimental group were subjected to static mechanical strain for 2h, whereas those in the control group underwent no strain. The biological roles of static strain and the p38/MMP-2 axis in BMMSC migration were evaluated by Transwell assays and western blotting by inhibiting p38 phosphorylation. RESULTS: There were significantly more Nestin+ cells in the bone calluses of the experimental group than in those of the control group. In addition, Nestin+/p-p38+ cell numbers were significantly higher in the experimental group than in the control group, indicating that static strain activated p38 signaling in BMMSCs in vivo. In accordance with in vivo results, static strain in vitro stimulated phosphorylation of p38 in BMMSCs. Furthermore, expression of MMP-2 was elevated in BMMSCs under static strain compared with the control, and strain-induced MMP-2 expression was abolished by inhibition of p38 phosphorylation in BMMSCs. Moreover, Transwell assay results showed that static strain promoted BMMSC migration, which was abolished by inhibition of p38 phosphorylation. CONCLUSIONS: The present study demonstrated that static strain can promote the migration ability of BMMSCs via p38/MMP-2 signaling. To the best of our knowledge, this study is the first report demonstrating that the p38/MMP-2 axis governs BMMSC migration under static mechanical strain.

Molecular mechanisms of water table lowering and nitrogen deposition in affecting greenhouse gas emissions from a Tibetan alpine wetland.

Rapid climate change and intensified human activities have resulted in water table lowering (WTL) and enhanced nitrogen (N) deposition in Tibetan alpine wetlands. These changes may alter the magnitude and direction of greenhouse gas (GHG) emissions, affecting the climate impact of these fragile ecosystems. We conducted a mesocosm experiment combined with a metagenomics approach (GeoChip 5.0) to elucidate the effects of WTL (-20 cm relative to control) and N deposition (30 kg N ha-1  yr-1 ) on carbon dioxide (CO2 ), methane (CH4 ) and nitrous oxide (N2 O) fluxes as well as the underlying mechanisms. Our results showed that WTL reduced CH4 emissions by 57.4% averaged over three growing seasons compared with no-WTL plots, but had no significant effect on net CO2 uptake or N2 O flux. N deposition increased net CO2 uptake by 25.2% in comparison with no-N deposition plots and turned the mesocosms from N2 O sinks to N2 O sources, but had little influence on CH4 emissions. The interactions between WTL and N deposition were not detected in all GHG emissions. As a result, WTL and N deposition both reduced the global warming potential (GWP) of growing season GHG budgets on a 100-year time horizon, but via different mechanisms. WTL reduced GWP from 337.3 to -480.1 g CO2 -eq m-2 mostly because of decreased CH4 emissions, while N deposition reduced GWP from 21.0 to -163.8 g CO2 -eq m-2 , mainly owing to increased net CO2 uptake. GeoChip analysis revealed that decreased CH4 production potential, rather than increased CH4 oxidation potential, may lead to the reduction in net CH4 emissions, and decreased nitrification potential and increased denitrification potential affected N2 O fluxes under WTL conditions. Our study highlights the importance of microbial mechanisms in regulating ecosystem-scale GHG responses to environmental changes.

UV radiation is the primary factor driving the variation in leaf phenolics across Chinese grasslands.

Due to the role leaf phenolics in defending against ultraviolet B (UVB) under previously controlled conditions, we hypothesize that ultraviolet radiation (UVR) could be a primary factor driving the variation in leaf phenolics in plants over a large geographic scale. We measured leaf total phenolics, ultraviolet-absorbing compounds (UVAC), and corresponding leaf N, P, and specific leaf area (SLA) in 151 common species. These species were from 84 sites across the Tibetan Plateau and Inner Mongolian grasslands of China with contrasting UVR (354 vs. 161 mW/cm(2) on average). Overall, leaf phenolics and UVAC were all significantly higher on the Tibetan Plateau than in the Inner Mongolian grasslands, independent of phylogenetic relationships between species. Regression analyses showed that the variation in leaf phenolics was strongly affected by climatic factors, particularly UVR, and soil attributes across all sites. Structural equation modeling (SEM) identified the primary role of UVR in determining leaf phenolic concentrations, after accounting for colinearities with altitude, climatic, and edaphic factors. In addition, phenolics correlated positively with UVAC and SLA, and negatively with leaf N and N: P. These relationships were steeper in the lower-elevation Inner Mongolian than on the Tibetan Plateau grasslands. Our data support that the variation in leaf phenolics is controlled mainly by UV radiation, implying high leaf phenolics facilitates the adaptation of plants to strong irradiation via its UV-screening and/or antioxidation functions, particularly on the Tibetan Plateau. Importantly, our results also suggest that leaf phenolics may influence on vegetation attributes and indirectly affect ecosystem processes by covarying with leaf functional traits.