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A series of organogelators based on L-phenylalanine has been synthesized and their gelation properties in various organic solvents were investigated. The results showed that these organogelators were capable of forming stable thermal and reversible organogels in various organic solvents at low concentrations, and the critical gel concentration (CGC) of certain solvents was less than 1.0 wt%. Afterward, the corresponding enthalpies (&Delta, Hg) were extracted by using the van &rsquo, t Hoff equation, as the gel&ndash, sol temperature (TGS) was the function of the gelator concentration. The study of gelling behaviors suggested that L-phenylalanine dihydrazide derivatives were excellent gelators in solvents, especially BOC&ndash, Phe&ndash, OdHz (compound 4). The effects of the solvent on the self-assembly of gelators were analyzed by the Kamlet&ndash, Taft model, and the gelation ability of compound 4 in a certain organic solvent was described by Hansen solubility parameters and a Teas plot. Morphological investigation proved that the L-phenylalanine dihydrazide derivatives could assemble themselves into an ordered structure such as a fiber or sheet. Fourier-transform infrared spectroscopy (FTIR) and hydrogen nuclear magnetic resonance (1H NMR) studies indicated that hydrogen bonding, &pi, &ndash, &pi, stacking, and van der Waals forces played important roles in the formation of a gel.

This paper studies the problem of spatial linkage synthesis for motion generation from the perspective of extracting geometric constraints from a set of specified spatial displacements. In previous work, we have developed a computational geometric framework for integrated type and dimensional synthesis of planar and spherical linkages, the main feature of which is to extract the mechanically realizable geometric constraints from task positions, and thus reduce the motion synthesis problem to that of identifying kinematic dyads and triads associated with the resulting geometric constraints. The proposed approach herein extends this data-driven paradigm to spatial cases, with the focus on acquiring the point-on-a-sphere and point-on-a-plane geometric constraints which are associated with those spatial kinematic chains commonly encountered in spatial mechanism design. Using the theory of kinematic mapping and dual quaternions, we develop a unified version of design equations that represents both types of geometric constraints, and present a simple and efficient algorithm for uncovering them from the given motion.

As one of the most aggressive malignancies, cervical cancer (CC) which mainly affects females has high risks of relapse and death. Long non-coding RNA (lncRNA) RP11-552M11.4 is verified to promote the progression of ovarian cancer; nevertheless, its role and the probable molecular mechanisms in CC remain unclear until the present study. Herein, we unveiled that the expression of lncRNA RP11-552M11.4 tested by qRT-PCR was enhanced in tumor tissues compared with the para-carcinoma tissues and related to FIGO Stage, lymph node metastasis, vascular invasion and distant metastasis in CC. Additionally, CC patients with high lncRNA RP11-552M11.4 level suffered from poor clinical outcomes. Moreover, silenced lncRNA RP11-552M11.4 restrained cell proliferation, migration and invasion in CC cells. Subsequently, the mechanistic studies revealed that lncRNA RP11-552M11.4 functioned as a ceRNA of ATF1 in CC by acting as the endogenous sponge for miR-3941, which was identified as a tumor suppressor in CC. Moreover, both miR-3941 inhibition and ATF1 overexpression restored the impacts of inhibited lncRNA RP11-552M11.4 on cellular processes in CC cells. Our observations elucidated the carcinogenic role of lncRNA RP11-552M11.4 in CC was mediated through miR-3941/ATF1 axis, giving a new insight into the effective target for the treatment and prognosis of cervical cancer.

Contents The aim of this study was to investigate the effects of donor cells’ sex on nuclear transfer efficiency and telomere length of cloned goats from adult skin fibroblast cells. The telomere length of somatic cell cloned goats and their offspring was determined by measuring their mean terminal restriction fragment (TRF) length. The result showed that (i) reconstructed embryos with fibroblast cells from males Boer goats obtained significantly higher kids rate and rate of live kids than those of female embryos and (ii) the telomere lengths of four female cloned goats were shorter compared to their donor cells, but five male cloned goats had the same telomere length with their donor cells, mainly due to great variation existed among them. The offspring from female cloned goats had the same telomere length with their age-matched counterparts. In conclusion, the donor cells’ sex had significant effects on nuclear transfer efficiency and telomere lengths of cloned goats.

mTOR activation is commonly caused by oncogenic mutations in RAS/RAF/MAPK and PI3K/AKT pathways, and promotes cancer progression and therapeutic resistance. However, mTOR inhibitors show limited single agent efficacy in patients. mTOR inhibitors suppress tumor cell growth and angiogenesis, and have recently been shown to induce death receptor/FADD-dependent apoptosis in colon cancers. Using a panel of BRAF V600E and WT colorectal cancer cell lines and in vitro selected resistant culture, and xenograft models, we demonstrate here that BRAFV600E confers resistance to mTOR inhibitors. Everolimus treatment disrupts the S6K1-IRS-2/PI3K negative feedback loop, leading to BRAF V600E-dependent activation of ERK and Mcl-1 stabilization in colon cancer cells, which in turn blocks the crosstalk from the death receptor to mitochondria. Co-treatment with inhibitors to Mcl-1, PI3K, RAF or MEK restores mTOR inhibitor-induced apoptosis by antagonizing Mcl-1 or abrogating ERK activation in BRAFV600E cells. Our findings provide a rationale for genotype-guided patient stratification and potential drug combinations to prevent or mitigate undesired activation of survival pathways induced by mTOR inhibitors.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive leukemia that is primarily caused by aberrant activation of the NOTCH1 signaling pathway. Recent studies have revealed that posttranslational modifications, such as ubiquitination, regulate NOTCH1 stability, activity, and localization. However, the specific deubiquitinase that affects NOTCH1 protein stability remains unestablished. Here, we report that ubiquitin-specific protease 7 (USP7) can stabilize NOTCH1. USP7 deubiquitinated NOTCH1 in vivo and in vitro, whereas knockdown of USP7 increased the ubiquitination of NOTCH1. USP7 interacted with NOTCH1 protein in T-ALL cells, and the MATH and UBL domains of USP7 were responsible for this interaction. Depletion of USP7 significantly suppressed the proliferation of T-ALL cells in vitro and in vivo, accompanied by downregulation of the NOTCH1 protein level. Similarly, pharmacologic inhibition of USP7 led to apoptosis of T-ALL cells. More importantly, we found that USP7 was significantly upregulated in human T-ALL cell lines and patient samples, and a USP7 inhibitor exhibited cell cytotoxicity toward primary T-ALL cells, indicating the clinical relevance of these findings. Overall, our results demonstrate that USP7 is a novel deubiquitinase that stabilizes NOTCH1. Therefore, USP7 may be a promising therapeutic target in the currently incurable T-ALL., Cancer pathology: Cutting a lifeline of lymphoblastic leukemia Inhibiting the protein ubiquitin-specific protease 7 (USP-7) may offer a treatment option for patients with T-cell acute lymphoblastic leukemia (T-ALL), an incurable and aggressive cancer. T-ALL is primarily caused by aberrant activation of the protein NOTCH1, which regulates gene expression relating to cell survival, proliferation, and metastasis. Ying-Li Wu, from China’s Shanghai Jiao Tong University School of Medicine, and a team of researchers report that, in their experiments, USP-7 prevented the breakdown of NOTCH1 by removing the protein tag that would normally signal it for degradation. In addition, USP-7 expression was upregulated in all human T-ALL cells and samples tested. Inhibition of USP-7 suppressed the proliferation of T-ALL cells in vitro and in vivo. These results reveal the role of USP-7 in the pathophysiology of T-ALL and offer USP-7 as a promising potential therapeutic target.

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) are prone to anoikis after single cell dissociation. The small molecule, Y-27632 is known to increase survival of hESCs and hiPSCs by inhibiting the Rho-associated protein kinase (ROCK). However, the underlying mechanisms are still unclear. Here, we thoroughly screened small molecules to investigate the adhesion and survival of hESCs in adherent culture. Y-27632 provided higher adhesion and survival of hESCs by increased cell migration and preventing cell blebbing in single dissociated cells. The combination of Matrigel with poly-d-lysine increased the attachment and survival of dissociated cells via actin filament and microtubule spreading in Y-27632-treated cells. Although Y-27632 prevented apoptosis by suppressing actin filament contraction, microtubule bundling, and blebbing, prolonged Y-27632 treatment presented a different morphology in the attached growing hESC colony. It induced apoptosis of cells by promoting cytoplasmic spread, E-cadherin structural change, and increased detachment. It also induced actin cytoskeleton disruption, combined with microtubule and intermediate filament elongation. For optimal hPSC culture, our research suggests that Y-27632 should be removed shortly after passaging. Keywords: Cytoskeletal changes, ROCK inhibitor, Apoptosis, Human embryonic stem cells, Adhesion, Extracellular matrices

When designing linkage mechanisms for motion synthesis, many examples have shown that the optimal kinematic constraint on the task motion contains too large deviation to be approximately viewed as a single rotational or translational pair. In this paper, we seek to adopt our previously established motion synthesis framework for the design of cam-linkages for a more accurate realization, while still maintaining a 1-degree-of-freedom (DOF) mechanism. To determine a feasible cam to lead through the task motion, first a kinematic constraint is identified such that a moving point on the given motion traces a curve that is algebraically closest to a circle or a line. This leads to a cam with low-harmonic contour curve that is simple and smooth to avoid the drawbacks of cam mechanisms. Additional constraints could also be imposed to specify the location and/or size of the cam linkages. An example of the design of a lower-limb rehabilitation device has been presented at the end of this paper to illustrate the feasibility of our approach. It is shown that our design could lead the user through a normal walking motion.

To better understand the function of oar-miR-150 in the ovine granulosa cells (GCs) during the estrus cycle, the five turpan sheep were selected for detection of the expression of oar-miR-150 in follicular and luteal ovaries, respectively. Then the granulosa cells treated with oar-miR-150 mimics or negative control (NC) were analyzed by qPCR to assess the expression of genes involved in steroidogenic and apoptosis. Expression of oar-miR-150 was increased in follicular phase compared with that in luteal ovaries. STAR was a target gene of miR-150 with oar-miR-150 mimic or inhibitor and luciferase reporter assay. Overexpression of oar-miR-150 promoted expression of bax, bcl2 and Casp3 expression and declined the expression of STAR, Cyp11a1 and HSD3B1 in vitro. Taken together, these results demonstrate that the oar-miR-150 promote GCs apoptosis through modifying the expression of genes involved in progesterone synthesis.

Mitochondrial function and morphology are dynamically regulated by fusion and fission. Heme oxygenase-1 (HO-1), which may be upregulated by protein kinase C-α (PKC-α), improves mitochondrial dynamics by controlling the balance between fusion and fission in vivo and in vitro. However, whether the PKC-α/HO-1 signaling pathway is one of the underlying mechanisms in adjusting mitochondrial dynamics in lipopolysaccharide (LPS)-activated macrophages has remained elusive. To explore this, NR8383 cells were pre-treated with PKC-α inhibitor Go6976 or PKC-α activator phorbol-12-myristate-13-acetate for 30 min and then stimulated with LPS for 24 h. Next, the expression of PKC-α, HO-1, mitofusin 1 (Mfn1) and mitofusin 2 (Mfn2), optic atrophy 1 (OPA1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1) was detected to evaluate the possible implication of the PKC-α/HO-1 signaling pathway in the LPS-induced NR8383 cells. The results indicated that activation of the PKC-α/HO-1 signaling pathway increased superoxide dismutase activities and the respiratory control ratio (RCR), decreased the levels of malondialdehyde, reactive oxygen species (ROS), Drp1 and Fis1, and simultaneously enhanced the levels of Mfn1, Mfn2 and OPA1. In contrast, the PKC-α inhibitor decreased the expression of RCR, Mfn1, Mfn2 and OPA1, and increased the expression of MDA and ROS in NR8383 cells. The results suggest that activation of the PKC-α/HO-1 signaling pathway is necessary for the balance of mitochondrial dynamics and oxidative stress in macrophages, which provides clues for probing novel strategies against the detrimental effects of sepsis and other disease states.

2-Methyl-2-butanol (MBT) is a chemical compound from the group of alcohols more specifically pentanols, which has shown an excellent anti-cancer activity in our previous study. However, its mechanism of action remains unclear. The present study was designed to investigate the anti-cancer effect of MBT on human retinoblastoma cells. The results showed that the use of MBT leads to HXO-RB44 cell death but is cytotoxic to normal cells at higher concentrations. It showed a dose- as well as a time-dependent inhibition of HXO-RB44 cells. P27 is a cell cycle inhibitory protein, which plays an important role in cell cycle regulation whereas cyclin-B1 is a regulatory protein involved in mitosis. MBT increased the cell cycle arrest in a dose-dependent manner by augmenting p27 and reducing cyclin B1 expression. Moreover, it also accelerated apoptosis, increased light chain-3 (LC-3) conversion in a dose-dependent manner, and helped to debulk cancerous cells. LC3 is a soluble protein, which helps to engulf cytoplasmic components, including cytosolic proteins and organelles during autophagy from autophagosomes. In order to verify the effect of MBT, bafilomycin A1, an autophagy inhibitor, was used to block the MTB-induced apoptosis and necrosis. Additionally, a specific Akt agonist, SC-79, reversed the MBT-induced cell cycle arrest and autophagy. Thus, from the present study, it was concluded that MBT induced cell cycle arrest, apoptosis and autophagy through the PI3K/Akt pathway in HXO-RB44 cells.

This paper studies the problem of spherical four-bar motion synthesis from the viewpoint of acquiring circular geometric constraints from a set of prescribed spherical poses. The proposed approach extends our planar four-bar linkage synthesis work to spherical case. Using the image space representation of spherical poses, a quadratic equation with ten linear homogeneous coefficients, which corresponds to a constraint manifold in the image space, can be obtained to represent a spherical RR dyad. Therefore, our approach to synthesizing a spherical four-bar linkage decomposes into two steps. First, find a pencil of general manifolds that best fit the task image points in the least-squares sense, which can be solved using singular value decomposition (SVD), and the singular vectors associated with the smallest singular values are used to form the null-space solution of the pencil of general manifolds; second, additional constraint equations on the resulting solution space are imposed to identify the general manifolds that are qualified to become the constraint manifolds, which can represent the spherical circular constraints and thus their corresponding spherical dyads. After the inverse computation that converts the coefficients of the constraint manifolds to the design parameters of spherical RR dyad, spherical four-bar linkages that best navigate through the set of task poses can be constructed by the obtained dyads. The result is a fast and efficient algorithm that extracts the geometric constraints associated with a spherical motion task, and leads naturally to a unified treatment for both exact and approximate spherical motion synthesis.

Background Acemannan is a bioactive polysaccharides promoting tissue repair. However, the roles of acemannan in skin wound healing and the underlying molecular mechanisms are largely unclear. Objective The goal of this study is to investigate the positive role of acemannan in cutaneous wound healing and its mechanism. Methods Mouse skin wound model and skin primary fibroblasts were used to demonstrate the positive effect of acemannan on cutaneous wound healing. The expressions of cell proliferation nuclear antigen ki-67, cyclin D1 and activity of AKT/mTOR signaling were analyzed in acemannan-treated fibroblasts and mice. Rapamycin and AKT inhibitor VIII were used to determine the key role of AKT/mTOR signaling in acemannan-promoting cutaneous wound healing. Results We found that acemannan significantly accelerated skin wound closure and cell proliferation. Acemannan promoted the expression of cyclin D1 in cultured fibroblasts, which was mediated by AKT/mTOR signal pathway leading to enhanced activity of the eukaryotic translation initiation factor-4F (eIF4F) and increased translation of cyclin D1. In contrast, pharmaceutical blockade of AKT/mTOR signaling by mTOR inhibitor rapamycin or AKT inhibitor VIII abolished acemannan-induced cyclin D1 translation and cell proliferation. In vivo studies confirmed that the activation of AKT/mTOR by acemannan played a key role in wound healing, which could be reversed by rapamycin. Conclusion Acemannan promoted skin wound healing partly through activating AKT/mTOR-mediated protein translation mechanism, which may represent an alternative therapy approach for cutaneous wound.

Ligustrazine (TMP) has recently been used for the treatment of various cancers. However, its exact mechanisms of action, particularly the functions and the mechanisms of Ligustrazine in human hormone-refractory prostate cancer (HRPC), have not yet been extensively studied. Recently, our findings suggest that Ligustrazine dose- and time-dependently inhibits the growth of HRPC cells by reducing their proliferation and promoting apoptosis. Interestingly, the treatment of hormone-refractory prostate cancer (PC-3) cells with Ligustrazine results in a significant inhibition of the activation of mTOR and related downstream targets, which are critical for cell growth. Furthermore, pull-down assays with 7-methyl- GTP Sepharose 4B beads indicate that Ligustrazine reduces the available eIF4E for translation initiation. Accordingly, the results from the translation assay using a luciferase reporter system further demonstrate that Ligustrazine indeed inhibits cap-dependent translation. In addition, the transient overexpression of eIF4E or MNK1 prevents the Ligustrazine-induced inhibition of proliferation and confers significant protection against Ligustrazine-induced apoptosis. Therefore, the present study provides evidences that Ligustrazine may be a candidate for therapeutic reagent for the treatment of HRPC and certifies that Ligustrazine modulates the availability of eIF4E mainly through the mTOR and MEK/ERK signaling pathways to inhibit cap-dependent translation. Taken together, our results indicate that the inhibition of cap-dependent translation is likely an essential mechanism in Ligustrazine-induced apoptosis.

We report the synthesis of rod-like hydroxyapatite nanopowder from waste eggshells (decomposed to CaO) and phosphoric acid at room temperature without pH control. The procedure involves the use of a syringe pump to control the addition rate of the phosphoric acid in order to maintain a stable pH. Results revealed that pure, rod-like, and nano-size hydroxyapatite particles were produced at various addition rates. Moreover, the sample obtained from 200 ml/h addition rate exhibited large surface area (160.1 m2/g) and pore volume (1.0 cm3/g). Consequently, the prepared hydroxyapatite and that previously produced as adsorbents showed fast adsorption rates (10 min) as well as very high maximum adsorption (qm = 994.5 mg/g and 1000.5 mg/g, respectively) for Pb2+ in aqueous solution. Furthermore, the adsorption process revealed a pseudo-second-order reaction model and the equilibrium adsorption could be described by Langmuir model.

Although efficient nonsurgical transfer of embryos in mice would provide many advantages over a surgical method, the low success rate of nonsurgical transfer has hampered its acceptance and use. Here, a plastic catheter was used to mimic embryo transfer process and then the transfer efficiency was evaluated by intrauterine trypan blue dye dispersion. Also 3.5-day blastocysts from natural pregnant mice were transferred through cervix into uterine horns. The results show that 70.9% of CD-1 mouse 3.5-day blastocysts transferred into unilateral uterine horns of pseudopregnant 2.5-day recipients can be developed to live newborns, and an efficient mouse nonsurgical embryo transfer technique was established. The technique was simple, rapid, inexpensive, unlikely to get contaminated, ethical and do not need specialized apparatus, and can completely replace surgical embryo transfer techniques. Moreover, the mouse nonsurgical embryo transfer technique provides a research model for human and other large animal embryo transfer.为了提高非手术性小鼠胚胎移植技术的成功率，利用塑料移植导管模拟非手术胚胎移植过程，通过观察染料在子宫角的分布而评估胚胎移植效果，并在此基础上将自然妊娠3.5 d 的小鼠囊胚经子宫颈移植受体小鼠。结果表明：将CD-1 小鼠囊胚移植假孕2.5 d 小鼠单侧子宫角，平均70.9%的胚胎能够发育至成活新生仔鼠，建立了高效非手术性小鼠胚胎移植技术。该方法简便快捷、不易污染、费用低，无需专业的手术器械，且符合实验动物伦理原则，完全可以取代手术法胚胎移植技术，更重要的是，它为人类和其他大动物的胚胎移植提供了研究模型。.

Ovarian function, the control of which is predominantly integrated by the hypothalamic-pituitary-ovarian axis, is pivotal for maintaining reproductive efficiency in sheep. To understand the regulatory mechanism of the follicular-luteal phase transition in prolific sheep, the genome-wide expression patterns of microRNAs and genes in ovarian tissue of Small Tail Han sheep were examined during the follicular and luteal phases of the oestrous cycle. Differentially expressed genes (n = 450) were identified and a total of 139 known and 72 novel miRNAs were identified in the two libraries. It is suggested that differential abundance of miR-200a, 200b and 200c may have an important role in the follicular-luteal transition. A miRNA-regulated gene expression network was created for exploring the regulation of the follicular-luteal transition and quantitative real-time PCR verified the reliability of the RNA-seq data.

We provide a cloud manufacturing based manufacturing planning framework for small and medium-sized enterprises. Manufacturing planning is conducted by separate units in the cloud instead of in corporations or manufacturing platforms. Disorders can be removed by the adoption of our newly-introduced units. To retain the workability of our new framework, three assumptions are imposed. A concrete case on process planning and scheduling is used for illustration of the necessity of our assumptions and operational mechanism of our design. Finally, a preliminary discuss on how intellect resources as well as small and medium-sized enterprises are involved to create a sustainable environment for small and medium-sized enterprises is placed.

Drug resistance to tyrosine kinase inhibitors (TKIs) is currently a clinical problem of chronic myelogenous leukemia (CML). Bcr-Abl protein depletion is considered as a way to overcome drug resistance to TKIs. In our study, Chk1 inhibitors, AZD7762 and MK-8776, had strong antitumor effects on CML cell line KBM5 and imatinib-resistant form KBM5T315I. Moreover, Chk1 inhibitors showed a strong cytotoxic effect on leukemia cells from primary CML and imatinib-resistance CML patients, but low cytotoxic effect on normal human mononuclear cells. Then, we found that Chk1 inhibitors induced apoptosis and increased DNA damage in CML cell lines with the degradation of the Bcr-Abl protein. Using the proteasome inhibitor and an immunoprecipitation assay, we found that Chk1 inhibitors triggered the degradation of Bcr-Abl through ubiquitination, which is depending on E3 ubiquitin ligase CHIP. At last, MK-8776 showed a significant tumor-suppressive effect of KBM5T315I cell in xenograft tumor models. Taking together, these findings suggest that targeting Chk1 may overcome TKIs resistance for the treatment of CML.

This paper deals with the classic problem of the synthesis of planar linkages for path generation. Based on the Fourier theory, the task curve and the synthesized four-bar coupler curve are regarded as the same curve if their Fourier descriptors match. Using Fourier analysis, a curve must be given as a function of time, termed a parametrization. In practical applications, different parametrizations can be associated with the same task and coupler curve, respectively; however, these parametrizations are Fourier analyzed to different Fourier descriptors, thus resulting in the mismatch of the task and coupler curve. In this paper, we present a parametrization-invariant method to eliminate the influence of parametrization on the values of Fourier descriptors by unifying given parametrizations to the arc length parametrization; meanwhile, a new design space decoupling scheme is introduced to separate the shape, size, orientation, and location matching of the task and four-bar curve, which leads naturally to an efficient synthesis approach.

Heat-labile enterotoxin (LT) can cause animal enteritis and diarrhea. However, the possible association of LT with embryo survival in pregnant animals and the mechanisms involved remain unknown. To investigate the effects of LT on embryo survival, we treated mouse early embryos in vitro and pregnant mice in vivo with recombinant LT. LT significantly decreased mouse embryo survival, and induced IFN-γ, IL-2 and IL-1β production in the serum and placental tissue. LT also triggered IL-1β release from LPS-primed microphages, suggesting LT can activate inflammasomes. To determine the pathway involved in LT-induced inflammasome activation, small interfering RNAs were used to knockdown NLRP3 and ASC, the key components of NLRP3 inflammasome pathway. Ablation of NLRP3 and ASC abolished LT-induced IL-1β release, confirming the involvement of NLRP3 inflammasome. By comparing two subunits of LT, only LTA but not LTB subunit was identified to activate the NLRP3 inflammasome.

Objective Although most embryologists load air bubbles into the catheter along with embryos during embryo transfer, the effects of these air bubbles on embryo transfer success rate are not clear. Study design Air bubbles were nonsurgically injected into unilateral uterine horns of mice to demonstrate the negative effects of intrauterine air bubbles on embryonic development. Results Our data showed that when air bubbles are nonsurgically injected into unilateral uterine horns of pregnant 4 days mice the litter size is significantly decreased. Four days after the introduction of air, abnormal decidua and dead conceptuses were detected in the uterine horns receiving the air bubbles. In addition, intrauterine air also significantly impaired murine embryo transfer success rates, and induced an increase in endometrial capillary permeability and decidualization in mice on day 4 of pseudopregnancy. These results strongly indicated that the air bubbles loaded into embryo transfer catheters to bracket the embryo-containing medium may have negative effect on embryonic implantation and development. Conclusions Intrauterine air impaired murine embryonic postimplantation development, and this provided some clues for improving embryo transfer techniques in human.

We employed a high-pressure solid–solid preparation technique in CO2 to synthesize carbonate hydroxyapatite (CHAp) powders from solid calcium and phosphate precursors. The synthesis parameters were investigated and optimized to determine the best precursor and optimal synthesis conditions. The results showed that the technique is green and that the biogenic calcium from eggshells in supercritical CO2 yielded B-type CHAp with desirable and recommended carbonate contents (4.1–5.1 wt %). The biogenic calcium source also yielded product with excellent surface area (212.4m2/g) and pore volume (0.65 cm3/g) compared to the synthetic calcium source, which exhibited high ibuprofen loading (270.0 mg/g) and enhanced dissolution (100% in 6 h) in simulated gastric fluid. These results indicate that the CHAp has potential for medical and other related applications. Mechanism study revealed that the procedure is a self-catalytic reaction triggered by water in the reactants with short time, which can overcome the disadvantages of existing techniques commendably.

The transforming growth factor-β (TGF-β) signaling pathway promotes tissue fibrosis and scarring through SMAD (small mothers against decapentaplegic)-dependent and SMAD-independent mechanisms. However, inhibition of SMAD-mediated signal transduction alone induces an excessive inflammatory response that impairs the antifibrotic effects of TGF-β inhibitors. In this study, we designed and characterized a dual-functional transcription activator protein 1 (AP-1) and SMAD decoy oligodeoxynucleotide, antifibrosis oligodeoxynucleotide 4 (AFODN4) in vitro and in vivo. AFODN4 binds directly to recombinant AP-1 and SMAD with high affinity. AFODN4 significantly inhibited the DNA-binding and transcriptional activities of both AP-1 and SMAD, as well as the production of fibrotic mediators stimulated by TGF-β1 or TGF-β2 in L929 murine fibroblasts. Local administration of AFODN4 significantly inhibited fibrosis associated with acute dermal wounds in mice. Intriguingly, AFODN4 inhibited AP-1-mediated production of proinflammatory mediators, which can be caused by blockage of SMAD alone in vitro and in vivo. Collectively, these findings suggest that dual inhibition of SMAD and AP-1 signaling by AFODN4 is a useful strategy for the development of new antifibrotic agents.

In an earlier work, we have combined a curve fitting scheme with a type of shape descriptor, Fourier descriptor (FD), to develop a unified method to the synthesis of planar four-bar linkages for generation of both open and closed paths. In this paper, we aim to extend the approach to the synthesis of planar four-bar linkages for motion generation in an FD-based motion fitting scheme. Using FDs, a given motion is represented by two finite harmonic series, one for translational component of the motion and the other for rotational component. It is shown that there is a simple linear relationship between harmonic content of the rotational component and that of the translational component for a planar four-bar coupler motion. Furthermore, it is shown that the rotational component of the given motion identifies a subset of design parameters of a four-bar linkage including link ratios, while the translational component determines the rest of the design parameters such as locations of the fixed pivots. This leads naturally to a decomposed design space for four-bar mechanism synthesis for approximate motion generation.

The design of mechanisms that lead a rigid-body through a set of prescribed discrete poses is usually referred to as “motion synthesis”. In practical motion synthesis cases, aside of realizing a set of given poses, various types of geometric constraint conditions could also require to be satisfied, e.g. defining the coordinates of the center/circle points of dyad linkages, setting the ground line/coupler line for four-bar linkages, realization of additional task positions, etc. Some of these constraint conditions require to be realized exactly while others might allow approximation. To solve this mixed-exact-and-approximated problem, this paper proposed a kinematic-mapping-based approach, which builds on the previous work of the realization of an arbitrary number of approximated poses as well as up to four exact poses. We now have found that the aforementioned various types of constraint conditions could be converted to each other through a general linear constraint equation. Thus, those “approximated conditions” could be uniformly converted to several prescribed discrete poses so as to be formulated as a general approximated motion synthesis problem, which is actually a general quadratic surface fitting problem in kinematic-mapping space, while up to four “exact conditions” could be imposed as linear constraint equations to this surface fitting system such that they could be exactly realized. Through null-space analysis technique, both type and dimensions of the resulting optimal dyad linkages could be determined by the solution of this surface-fitting problem with constraints. These optimal dyads could then be implemented as different types of four-bar linkages or parallel manipulators.

Psoriatic keratinocytes express exaggerated levels of inflammatory cytokines, and show aberrant hyperproliferation and terminal differentiation in the pathogenesis of psoriasis. The antimicrobial protein hS100A7 (psoriasin) has been found highly expressed in psoriatic skin, but the mechanism and physiological function remain largely unknown. We observed that hS100A7 induces mature interleukin 1α (17kDa) expression in normal human epidermal keratinocytes, which is dependent on RAGE-p38 MAPK and calpain-1 as the inhibitors or knockdown of them completely decreased the expression of mature interleukin1α. Then, we proved mS100a7a15, mature IL-1α and calpain-1 were highly expressed in imquimod-induced psoriasis model and mouse IL-17a-neutralizing antibody treatment attenuated mS100a7a15 expression. At last, PD 151746 (calpain-1 inhibitor) treatment decreased epidermal thickness in imquimod-induced psoriasis model. Taken together, our results suggest that mature IL-1α induced by hS100A7 is via RAGE-p38 MAPK and calpain-1 pathway in keratinocyte and this mechanism may play an important role during psoriasis.

This paper deals with the problem of integrated joint type and dimensional synthesis of planar four-bar and six-bar linkages, which could contain both revolute (R) and prismatic (P) joints, for guiding through five specified task positions of the end-effector. In a recent work, we developed a simple algorithm for analyzing a set of given task positions to determine all feasible planar dyads with revolute and/or prismatic joints that can be used to guide through the given positions. This paper extends this algorithm to the integrated joint type and dimensional synthesis of Watt I and II and Stephenson I, II, and III six-bar linkages that contain both R- and P-joints. In the process, we developed a new classification for planar six-bar linkages according to whether the end-effector can be constrained by two dyads (type I), one dyad (type II), or no dyad (type III). In the end, we demonstrate this task-driven synthesis approach with three examples including a novel six-bar linkage for lifting an individual with age disability from seating position to standing position.

eIF4E, Eukaryotic Translation Initiation Factor 4E, regulates the rate limiting cap-binding step in mRNA translation. Oncogenic pathways converge to increase eIF4E expression, as well as phosphorylation of eIF4E (S209) and its inhibitor 4EBPs, which are believed to facilitate the translation of “weak'' mRNAs including cell type-specific and oncogenic targets. eIF4E phosphorylation, but not eIF4E, is dispensable for normal development. Pharmacologic inhibition of either p-eIF4E or p-4EBP has limited antitumor efficacy. Our current study aims to better define the role of p-eIF4E in colon cancer. We find that p-eIF4E, not eIF4E transcript or protein is elevated in human colon tumors and precursors. Knockin (KI) of phosphorylation-defective eIF4E mutant allele (4ES209A) in human colon cancer cells strongly suppresses Myc translation and cell growth under stress or in mice, while minimally impacting Cap binding or global translation. Surprisingly, p-eIF4E is found to be required to maintain p-4EBP1 levels, dissociate eIF4E from 4E-BP1, and stimulate glutamine metabolism via integrated stress response (ISR). Further, p-eIF4E and p-4EBP1 are highly elevated in the polyps in APC min/+ mice compared to adjacent normal intestinal crypts, and 4ES209A KI significantly inhibits intestinal polyposis and glutamine metabolism gene signatures. Finally, p-eIF4E renders CRC cells addicted to glutamine and glutaminolysis by preventing Myc- and ISR-driven metabolic crisis and cell death. In summary, we demonstrate that eIF4E phosphorylation is central to oncogenic translation, and is required to license Myc-driven metabolic adaptation and addiction via ISR-dependent transcriptional programs in colon cancer. These novel findings help reconcile the long-standing and context-dependent roles of ISR in cancer cell growth, death, plasticity and heterogeneity, and provide new ways for its therapeutic exploitation. Citation Format: Hang Ruan, Xiangyun Li, Brian Leibowitz, Jingshan Tong, Xinghua Lu, Nahum Sonenberg, Xiang Xu, Lin Zhang, Jian Yu. eIF4E phosphorylation licenses Myc-driven metabolic and stress reprogramming [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2417.

Human embryonic stem (hES) cells hold great promise for application of human cell and tissue replacement therapy. However, the overwhelming majority of currently available hES cell lines have been directly or indirectly exposed to materials containing animal-derived components during their derivation, propagation, and cryopreservation. Unlike feeder-based cultures, which require the simultaneous growth of feeder and stem cells, resulting in mixed cell populations, stem cells grown on feeder-free systems are easily separated from the surface, presenting a pure population of cells for downstream applications. In this study, we have developed a novel method to expand hES cells in xeno-free, feeder-free conditions using 2 different matrices derived from xeno-free human foreskin fibroblasts (XF-HFFs). Using XF-HFF-derived extracellular matrix, together with 100 ng/mL recombinant bFGF-supplemented HEScGRO Basal Medium, long-term xeno-free expansion of hES cells is possible. Resulting hES cells were subjected to stringent tests and were found to maintain ES cell features, including morphology, pluripotency, stable karyotype, and expression of cell surface markers, for at least 20 passages. Xeno-free culturing practices are essential for the translation of basic hES cell research into the clinic. Therefore, the method presented in this study demonstrates that hES cells can be cultured in complete xeno-free conditions without the loss of pluripotency and furthermore, without the possibility of contamination from exogenous sources.

Colorectal cancers with mutant RAS/RAF are therapy refractory. Deregulated mRNA translation has become an emerging target in cancer treatment. We recently reported that mTOR inhibitors induce apoptosis via ER stress and the extrinsic pathway upon acute inhibition of the eIF4F complex in colon cancer cells and xenografts, while mutant BRAF600E leads to therapeutic resistance via ERK-mediated Mcl-1 stabilization. In this study, we demonstrated that several other translation inhibitors also activate ER stress and the extrinsic apoptotic pathway. Co-targeting translation and proteasome using the combination of Episilvestrol and Bortezomib promoted strong ER stress and rapid killing of colon cancer cells with mutant RAS/RAF in culture and mice. This combination led to marked induction of ER stress and ATF4/CHOP, followed by DR5- and BAX-dependent apoptosis, but unexpectedly with maintained or even increased levels of prosurvival factors such as p-AKT, p-4E-BP1, Mcl-1, and eiF4E targets c-Myc and Bcl-xL. Our study supports that targeting deregulated proteostasis is a promising approach for treating advanced colon cancer via induction of destructive ER stress that overcomes multiple resistance mechanisms associated with translation inhibition.

Since their derivation, human embryonic stem (hES) cells have been used for a variety of applications including developmental biology, pathology, chemical biology, genomics, and proteomics. However, their most important potential application is the generation of cells and tissues, which can be used for cell-based therapies. One of the main drawbacks of hES cell culture is that they are particularly sensitive to dissociation, which is required for passaging, expansion, cryopreservation, and other applications. Recently, it has been discovered that an inhibitor of Rho kinase (ROCKi; Y-27632) increases the survival rate of dissociated, single hES cells. This breakthrough has allowed new methods in hES cell culture to be developed, with the promise of increasing hES cell numbers into the realm of clinical relevance. In our studies demonstrating that ROCKi dramatically increases hES cell cryopreservation efficiency, we have observed that ROCKi treatment does not decrease hES cell's susceptibility to apoptosis. Rather, we hypothesize that ROCKi treatment desensitizes single hES cells to their environment reducing the odds that individual cells will undergo anoikis.

After slow freezing, the survival rate of human embryonic stem (hES) cells is poor and inconsistent. The aim of this study was to increase the freeze-thaw survival rate of hES cells by utilizing the ROCK inhibitor Y-27632. hES cell colonies were first treated with Y-27632, followed by collagenase IV and TrypLE Select dissociation whereupon small clumps were slow frozen using 90% Knockout serum replacement and 10% dimethyl sulfoxide. After thawing at 37 C, the clumps were cultured in medium supplemented with 10 microM Y-27632 for 1 day. Our results show that the use of Y-27632 significantly increases the survival of hES cells after thawing compared with that of the control group. Y-27632-treated freeze-thawed hES cells retain morphology, stable karyotype, expression of cell surface markers, and the potential to differentiate into derivatives of all three germ layers after long-term culture. We have concluded that conventional slow freezing with Y-27632 treatment is efficient and convenient for the cryopreservation of hES cells.

To investigate the relationship between the methylation status of imprinted gene H19 and the development of mice derived completely from Embryonic stem cells (ES) by tetraploid embryo complementation. The methylation status of two loci at 5'UTR region in imprinted gene H19 in normal adult control mice, 22 adult ES mice, 8 newborn dead ES mice and embryonic stem (ES) cells with different passage number were detected by using of methylation-sensitive restriction endonuclease-PCR technique. The results indicated that the methylation status of the imprinted gene H19 in adult ES mice were identical to that of adult control mice. However, some significant differences in the methylation status of the imprinted gene H19 were found among newborn dead ES mice, adult ES mice and normal adult control mice. Furthermore, the methylation status of the imprinted gene H19 in ES cells were probably different from that of adult ES mice and normal adult control mice.