The Influence of Deep Cryogenic Treatment (DCT) on the Microstructure Evolution and Mechanical Properties of TC4 Titanium Alloy.

Deep cryogenic treatment (-196 °C, DCT) is an emerging application that can make significant changes to many materials. In this study, DCT was applied to Ti6Al4V (TC4) titanium alloy, and we delved into an examination of the impact on its microstructural morphologies and mechanical properties. It was observed that DCT has a significant effect on the grain refinement of the TC4 titanium alloy base material. Obvious grain refinement behavior can be observed with 6 h of DCT, and the phenomenon of grain refinement becomes more pronounced with extension of the DCT time. In addition, DCT promotes the transformation of the ß phase into the α' phase in the TC4 titanium alloy base material. XRD analysis further confirmed that DCT leads to the transformation of the ß phase into the α' phase. The element vanadium was detected by scanning electron microscopy, and it was found that the ß phase inside the base material had transformed into the α' phase. It was observed that DCT has a positive influence on the hardness of the TC4 titanium alloy base material. The hardness of the sample treated with 18 h of DCT increased from 331.2 HV0.5 to 362.5 HV0.5, presenting a 9.5% increase compared to the sample without DCT. Furthermore, it was proven that DCT had little effect on the tensile strength but a significant impact on the plasticity and toughness of the base material. In particular, the elongation and impact toughness of the sample subject to 18 h of DCT represented enhancements of 27.33% and 8.09%, respectively, compared to the raw material without DCT.

Optimal planar leg geometry in robots and crabs for idealized rocky terrain.

Natural terrain is uneven so it may be beneficial to grasp onto the depressions or 'valleys' between obstacles when walking over such a surface. To examine how leg geometry influences walking across obstacles with valleys, we (1) modeled the performance of a two-linkage leg with parallel axis 'hip' and 'knee' joints to determine how relative segment lengths influence stepping across rocks of varying diameter, and (2) measured the walking limbs in two species of intertidal crabs,Hemigrapsus nudusandPachygrapsus crassipes, which live on rocky shores and granular terrains. We idealized uneven terrains as adjacent rigid hemispherical 'rocks' with valleys between them and calculated kinematic factors such as workspace, limb angles with respect to the ground, and body configurations needed to step over rocks. We first find that the simulated foot tip radius relative to the rock radius is limited by friction and material failure. To enable force closure for grasping, and assuming that friction coefficients above 0.5 are unrealistic, the foot tip radius must be at least 10 times smaller than that of the rocks. However, ratios above 15 are at risk of fracture. Second, we find the theoretical optimal leg geometry for robots is, with the distal segment 0.63 of the total length, which enables the traversal of rocks with a diameter that is 37% of the total leg length. Surprisingly, the intertidal crabs' walking limbs cluster around the same limb ratio of 0.63, showing deviations for limbs less specialized for walking. Our results can be applied broadly when designing segment lengths and foot shapes for legged robots on uneven terrain, as demonstrated here using a hexapod crab-inspired robot. Furthermore, these findings can inform our understanding of the evolutionary patterns in leg anatomy associated with adapting to rocky terrain.

Up-regulation of miR-27a promotes monocyte-mediated inflammatory responses in Kawasaki disease by inhibiting function of B10 cells.

Kawasaki disease (KD) is an acute systemic vasculitis and activation of monocytes plays a central role in the pathogenesis of it. B10 cells, a B cell subset with negative regulatory properties, are functionally identified by their ability to express cytoplasmic IL-10 after ex vivo stimulation. Here, we aimed to explore the functional role of B10 cells during monocyte-mediated inflammatory responses in KD, as well as elucidate the underlying microRNA (miRNA)-mediated regulatory mechanisms. Expression of IL-10 by each group of B cells (total B cells, transitional B cells, naïve B cells, and memory B cells) and inhibition of monocyte-derived TNF-α by activated B cells were measured by flow cytometry. Expression of miRNAs (miR-21-3p, miR-98-5p/3p, miR-27a-3p, let7b-5p, and miR-1423p/5p) that affect IL-10 levels in B cells was quantitated by real-time PCR. The relationship between IL-10 and these miRNAs was examined by multivariate analysis. MiR-mediated RNA interference in B cells was performed to investigate the role of miR-27a on expression of IL-10. The results showed expression of cytoplasmic IL-10 in B cell subsets from patients with KD was down-regulated. The inhibitory effect of B10 cells on production of TNF-α by monocytes from patients with KD was also compromised. The miR-27a-3p expression was markedly up-regulated during the acute phrase of KD, and it promoted monocyte-mediated TNF-α release by negatively regulating expression of cytoplasmic IL-10 within B cells in vitro. The data suggest up-regulated miR-27a in B cells from patients with KD may promote monocyte-mediated inflammatory responses by inhibiting the regulatory function of B10 cells.

[Role of miR-144-3p and its target gene in regulating osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro].

OBJECTIVE: To investigate the role of miR-144-3p in regulating osteogenic differentiation of bone marrow mesenchymal stem cells and predict its target genes. METHODS: Rat bone marrow mesenchymal stem cells (BMSCs) with induced osteogenic differentiation were examined for the expressions of Runx2, OCN and miR-144-3p. The effects of transfection with a miR-144-3p mimic or a miR-144-3p inhibitor were tested on the osteogenic differentiation of the BMSCs. The changes in the expressions of the predicted target of miR-144-3p in the BMSCs during induced osteogenic differentiation were examined using Western blotting and qRT-PCR. RESULTS: Rat BMSCs with induced differentiation into osteoblasts exhibited a progressive increase in the expressions of Runx2 and OCN (two markers of osteogenic differentiation), while the expression of miR-144-3p gradually decreased during the differentiation till reaching the lowest level at 21 days of induction. In rat BMSCs, transfection with the miR-144-3p mimic significantly decreased ALP activity (P < 0.05) wile transfection with the miR-144-3p inhibitor significantly increased ALP activity (P < 0.05) in rat BMSCs. Analysis based on miRanda, microRNA.org database and TargetScan suggested that Smad4 was the most likely target gene of miR-144-3p. The results of qRT-PCR showed no significant differences in expression levels of Smad4 among the cells with different treatments (P > 0.05), while Western blotting revealed a significantly decreased expression of Smad4 in the cells transfected with miR-144-3p mimics and an increased Smad4 expression in the cells transfected with the miR-144-3p inhibitor as compared with the control cells (P < 0.05). CONCLUSIONS: miR-144-3p participates in the regulation of osteogenic differentiation of rat BMSCs, and its inhibitory effect on osteogenic differentiation is achieved probably by decreasing the expression level of Smad4.

Identification of a novel de novo gain-of-function mutation of PIK3CD in a patient with activated phosphoinositide 3-kinase δ syndrome.

Activated phosphoinositide 3-kinase δ (PI3Kδ) syndrome is a newly defined and relatively common primary immunodeficiency, which is caused by heterozygous gain-of-function (GOF) mutations in PIK3CD or PIK3R1. Here, we report a novel de novo GOF mutation (c.1570 T > A, p.Y524N) in PIK3CD in a 6-year-old Chinese girl. The patient suffered recurrent sinopulmonary infection, bronchiectasis, lymphoproliferation, herpesvirus infection, and distinctive nodular lymphoid hyperplasia of mucosal surfaces. Immunological analysis revealed increased CD4+ T cell senescence and B cell immaturity. Further analysis revealed an increase in almost all CD4+ T cell subsets to varying degrees, including effector T cells and Treg cells. Increased levels of plasma T cell-related cytokines corroborated these results. Hyperactivation of the PI3Kδ-Akt-mTOR signaling pathway was also confirmed. Treatment with rapamycin ameliorated the lymphoproliferative immunodeficiency caused by hyperactivation of mTOR. These results expand genetic spectrum of APDS and will facilitate further study of the genotype-phenotype correlation in those with PIK3CD mutations.

[Expression and characterization of Coprinus cinereus peroxidase].

OBJECTIVE: The aim of our study is to express Coprinus cinereus peroxidase (CIP) in Pichia Pastori efficiently. METHODS: We synthesized CIP gene with P. pastori codon bias by our Gene Synthesis and site-specific mutagenesis platform, using DNAWorks 3.1 program to design and optimize primers. Then, we sequenced the PCR products, inserted the correct gene into expression vector pPICZαA and transformed the linearized pPICZαA-Cip DNA into P. pastori GS115. We integrated CIP gene into the genome of P. pastori, using the α-mating factor from Sacchoramyces cerevisiae as signal peptide to direct the secretion of the recombinant protein. To obtain transformants with high CIP activity, we checked transformants by nested PCR and stained 82 positive ones on YPD agar plate with 1000 mg/L Zeocin. Then, we got 6 transforments with high resistance to Zeocin and expressed them in small scale; the one exhibiting the highest activity was chosen as engineered strain and named CIP/Gs115. RESULTS: We purified CIP from culture medium after induction with ethanol, the maximum activity reached 487.5 U/mL on the 4th day. The purified CIP exhibited maximal activity at pH 5.0 and 25 degrees C with ABTS as substrate. The enzyme had 61.5% of the maximal activity at 45 degrees C and was stable below 40 degrees C. However, the stability was drastically reduced above 45 degrees C. The recombinant CIP remained stable between pH 4.5 and 6.5. We studied the substrate specificity on different substrates with the purified enzyme, and the optimal substrates were in the order of ABTS > 2, 6-Dimethoxyphenol > guaiacol > 2, 4-Dichlorophenol > phenol. CONCLUSION: The highly secretory expression of CIP and high special activity lay the good foundation for it' s industrial applications in waste water treatment, decolouration of dyestuffs.