Table1_Revealing phosphorylation regulatory networks during embryogenesis of honey bee worker and drone (Apis mellifera).XLSX

Protein phosphorylation is known to regulate a comprehensive scenario of critical cellular processes. However, phosphorylation-mediated regulatory networks in honey bee embryogenesis are mainly unknown. We identified 6342 phosphosites from 2438 phosphoproteins and predicted 168 kinases in the honey bee embryo. Generally, the worker and drone develop similar phosphoproteome architectures and major phosphorylation events during embryogenesis. In 24 h embryos, protein kinases A play vital roles in regulating cell proliferation and blastoderm formation. At 48–72 h, kinase subfamily dual-specificity tyrosine-regulated kinase, cyclin-dependent kinase (CDK), and induced pathways related to protein synthesis and morphogenesis suggest the centrality to enhance the germ layer development, organogenesis, and dorsal closure. Notably, workers and drones formulated distinct phosphoproteome signatures. For 24 h embryos, the highly phosphorylated serine/threonine-protein kinase minibrain, microtubule-associated serine/threonine-protein kinase 2 (MAST2), and phosphorylation of mitogen-activated protein kinase 3 (MAPK3) at Thr564 in workers, are likely to regulate the late onset of cell proliferation; in contrast, drone embryos enhanced the expression of CDK12, MAPK3, and MAST2 to promote the massive synthesis of proteins and cytoskeleton. In 48 h, the induced serine/threonine-protein kinase and CDK12 in worker embryos signify their roles in the construction of embryonic tissues and organs; however, the highly activated kinases CDK1, raf homolog serine/threonine-protein kinase, and MAST2 in drone embryos may drive the large-scale establishment of tissues and organs. In 72 h, the activated pathways and kinases associated with cell growth and tissue differentiation in worker embryos may promote the configuration of rudimentary organs. However, kinases implicated in cytoskeleton organization in drone embryos may drive the blastokinesis and dorsal closure. Our hitherto most comprehensive phosphoproteome offers a valuable resource for signaling research on phosphorylation dynamics in honey bee embryos.</p

The modified multiple platform method.

<p>Sleep deprivation was achieved by depriving the rats of paradoxical sleep. (A) Photos of circular narrow platforms with rats inside. (B) Photos of a grid floor with rats inside. The tank is filled with water up to approximately 1 cm from the surface of the platform or grid.</p

Comparison of remaining nutrient contents (fraction of initial nutrient content) among nutrient addition treatments in the late stage of decomposition.

<p>(a) and (b): S.S. leaf litter; (c) and (d): C.C. leaf litter. Significant differences (<i>P</i><0.05) among treatments are indicated by different letters. Error bars show SE (n = 5).</p

Mean decomposition rates (<i>k</i>) for S.S. and C.C. decomposed in the different nutrient addition treatments.

<p>Decomposition rate (<i>k</i>) and coefficients of determination (<i>R²</i>) are based on a single negative exponential model.</p><p>Significant differences (p<0.05) among treatments are indicated by different letters. Error bars show SE (n = 5).</p

Expression levels of p-ERK, ERK, MMP-1, MMP-3, and MMP-13 in the condyles.

<p>Western blot technique was used to examine the possible mechanism by which pathological alterations occur in the TMJ. (A) Comparison of the p-ERK, ERK, MMP-1, MMP-3, and MMP-13 protein levels in the different groups as determined by Western blot (WB). (B) Mean relative protein levels of p-ERK, ERK, MMP-1, MMP-3, and MMP-13 in different groups (n = 10 per group). Bars represent the mean and SD of each group. CON, control; CSD, chronic sleep deprivation; d, day. **<i>P</i><0.01, *<i>P</i><0.05.</p

Matrix Metalloproteinase Responsive Nanoparticles for Synergistic Treatment of Colorectal Cancer via Simultaneous Anti-Angiogenesis and Chemotherapy

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers worldwide, especially in developed countries. Although patients’ overall survival has been improved by either conventional chemotherapy or newly developed anti-angiogenesis treatment based on its highly vascularized feature, the relatively low therapeutic efficacy and severe side effects remain big problems in clinical practice. In this study, we describe an easy method to construct a novel matrix metalloproteinase-2 (MMP-2) responsive nanocarrier, which can load hydrophobic agents (camptothecin and sorafenib) with high efficiency to exert synergistic efficacy for CRC treatment. The drug-containing nanoparticles can particularly respond to the MMP-2 and realize the controlled release of payloads at the tumor site. Moreover, both in vitro and in vivo studies have demonstrated that this responsive nanoparticle exhibits much higher therapeutic efficacy than that of single antitumor agents or combined drugs coadministrated in traditional ways

Ultrastructure of the condyle visualized by scanning electron microscopy (SEM).

<p>SEM was used to observe the subtle ultrastructural alterations of the TMJ. Upper panel: Condylar fibrous articular surfaces of the U0126 rats at 7, 14, and 21 days of sleep deprivation. Middle panel: Condylar fibrous articular surfaces of the control group at 7, 14, and 21 days. Lower panel: Condylar fibrous articular surfaces of the CSD group at 7, 14, and 21 days of sleep deprivation (original magnification: ×2,500).</p

Table_1_Circadian Rhythm Protein Bmal1 Modulates Cartilage Gene Expression in Temporomandibular Joint Osteoarthritis via the MAPK/ERK Pathway.docx

The purpose of this study was to elucidate the role of the circadian gene Bmal1 in human cartilage and its crosstalk with the MAPK/ERK signaling pathway in temporomandibular joint osteoarthritis (TMJ-OA). We verified the periodical variation of the circadian gene Bmal1 and then established a modified multiple platform method (MMPM) to induce circadian rhythm disturbance leading to TMJ-OA. IL-6, p-ERK, and Bmal1 mRNA and protein expression levels were assessed by real-time RT-PCR and immunohistochemistry. Chondrocytes were treated with an ERK inhibitor (U0126), siRNA and plasmid targeting Bmal1 under IL-6 simulation; then, the cells were subjected to Western blotting to analyze the relationship between Bmal1 and the MAPK/ERK pathway. We found that sleep rhythm disturbance can downregulate the circadian gene BMAL-1 and improve phosphorylated ERK (p-ERK) and IL-6 levels. Furthermore, Bmal1 siRNA transfection was sufficient to improve the p-ERK level and aggravate OA-like gene expression changes under IL-6 stimulation. Bmal1 overexpression relieved the alterations induced by IL-6, which was consistent with the effect of U0126 (an ERK inhibitor). However, we also found that BMAL1 upregulation can decrease ERK phosphorylation, whereas ERK downregulation did not change BMAL1 expression. Collectively, this study provides new insight into the regulatory mechanism that links chondrocyte BMAL1 to cartilage maintenance and repair in TMJ-OA via the MAPK/ERK pathway and suggests that circadian rhythm disruption is a risk factor for TMJ-OA.</p

CORT and ACTH levels in serum.

<p>CORT, cortisol; ACTH, adrenocorticotropic hormone; CON, control; CSD, chronic sleep deprivation; d, days.</p><p>*<i>P</i><0.05, significantly different from the control group. Data are represented as the M ± SD of n = 10. M, mean; SD, standard deviation; n, sample size.</p><p>CORT and ACTH levels in serum.</p

MMP-1, 3, and 13 mRNA relative levels in the condylar cartilage.

<p>The 2^−ΔΔCt method was adopted with GAPDH as the reference gene.</p><p>CON, control; CSD, chronic sleep deprivation; chronic sleep deprivation with U0126 injection; d, days.</p><p>*<i>P</i><0.05, significantly different from the control group;</p>#<p><i>P</i><0.05, significantly different from the CSD group. Data are represented as the M ± s of n = 10. M, mean; SD, standard deviation; n, sample size.</p><p>MMP-1, 3, and 13 mRNA relative levels in the condylar cartilage.</p