Electrochemical tuning of a Cu3P/Ni2P hybrid for a promoted hydrogen evolution reaction.

Developing novel and high performance electrocatalysts for use in hydrogen evolution reactions (HER) as substitutes for noble metal based electrocatalysts is imperative and, so far, has been a challenge. Herein, a self-supported Cu3P/Ni2P hybrid on nickel foam (Cu3P/Ni2P@NF) is prepared by a simple galvanic replacement reaction coupled with phosphorization. Subsequently, Cu3P/Ni2P@NF is modified by conducting cyclic voltammetry scans in 0.5 M H2SO4 solution. Interestingly, after electrochemical tuning, the as-prepared Cu3P/Ni2P@NF exhibits significantly enhanced HER activity. Particularly, the resultant Cu3P/Ni2P@NF catalyst after 4000 cycles exhibits superior catalytic activity and long-term stability for HER with an overpotential of only 67 mV at the current density of 10 mA cm-2, and a low Tafel slope of 43.9 mV dec-1. The improved HER performance is attributed to the increased intrinsic activity of the Cu3P/Ni2P@NF with its optimized crystal and electronic structure, as well as an increased number of accessible active sites due to surface dissolution and recrystallization induced by electrochemical modification.

Bacterial Lipopolysaccharide Augmented Malignant Transformation and Promoted the Stemness in Prostate Cancer Epithelial Cells.

PURPOSE: To study bacterial lipopolysaccharide (LPS)-induced cancer stem-like transformation and to investigate the inhibitory effect of Trichostatin A (TSA) on the malignant transformation through targeting p-Stat3 signaling. METHODS: 2D, 3D, and serum-free suspension culture system were used to study LPS-induced malignant transformation in series malignant grade of prostate cancer (PCa) epithelial cells. Flow cytometry assay and RT-PCR were utilized to evaluate the CD44+CD133+ stem cell population, the expression of inflammatory cytokines and series tumor stemness biomarkers. Meanwhile, Western blot was used to analyze the alteration of cell signaling associated-molecules by treatment with TSA, an original antifungal antibiotic and a panel inhibitor of histone deacetylase. RESULTS: Our study found that LPS promoted the migration, invasion and stem-like tumoroshpere forming in multiple PCa cell lines including DU145, PC3, 22RV1, LNCaP. LPS also enriched CD44+CD133+ stem cell population and increased the expression of series tumor stemness biomarkers (e.g., CD44, CD133, SOX-2, α-intergrin, Nestin, etc.). TSA was found to prevent tumor cell migration, invasion and tumorosphere forming in DU145 and PC3 cells with increasing tumor suppressive Maspin and reducing both phosphorylation of Stat3 (p-Stat3) and pro-oncogene c-Myc expression in LPS-treated DU145 cells. Furthermore, blocking Stat3 signaling pathway by treatment with TSA and/or small molecule compound Stattic of an p-Stat3 inhibitor effectively abrogated LPS-induced tumorosphere forming with decrease of IL-6, IL-8 and stemness biomarkers CD44, SOX-2 expression. CONCLUSION: Our data demonstrated that the inflammatory agent of bacterial LPS augmented malignant transformation and promoted the cancerous stemness in PCa epithelial cells. TSA could prevent, at least in part, the LPS-induced malignant transformation by targeting p-Stat3/c-Myc signaling pathway and reducing inflammatory IL-6, IL-8. In addition, the assay of LPS-induced tumorosphere forming could serve as a simple and an easy handling method for targeting cancer stem cells drug screening in vitro in clinical practice.

Tumor Suppressive Maspin-Sensitized Prostate Cancer to Drug Treatment Through Negative Regulating Androgen Receptor Expression.

Overactivation of androgen receptor (AR)-mediated signal has been extensively implicated in prostate cancer (CaP) development, progression, and recurrence, which makes it an attractive therapeutic target. Meanwhile, as an endogenous inhibitor of histone deacetylase 1 (HDAC 1), tumor-suppressive mammary serine protease inhibitor (maspin) was reported to sensitize drug-induced apoptosis with a better therapeutic outcome in CaP, but the relationship between AR and maspin remains unclear. In the current study, treatment of 5'-Aza or MS-275/enzalutamide induced poly (ADP-ribose) polymerase (PARP) cleavage and p-H2A.X in CaP cells with an increase of maspin expression but a decrease of AR. Then, treatment with protease inhibitor MG132 did not rescue the above drug-induced loss of AR. In addition, modulation of maspin expression by gene recombinant or siRNA technology showed an inverse correlation between expression of maspin and AR, consequently affecting the AR-regulated downstream gene transcription (e.g., NKX3.1 and TMPRSS2). Bioinformatics analysis of the data extracted from the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) database also revealed an inverse correlation between low maspin expression and high AR level in advanced CaP. Furthermore, chromatin immunoprecipitation (ChIP) assay using anti-maspin antibody identified that a portion of AR promoter sequence was co-precipitated and presented in the immunoprecipitated complex. Finally, maspin-mediated repression of AR was induced by treatment of MS-275, which promoted enzalutamide treatment efficacy with decrease of prostate-specific antigen (PSA) expression in LNCaP and 22RV1 cells. Taken together, the data not only demonstrated maspin-mediated repression of AR to augment drug anti-tumor activity but also provided in-depth support for combination of HDAC inhibitors with AR antagonist in CaP therapy.

Precision Landing Test and Simulation of the Agricultural UAV on Apron.

Unmanned aerial vehicle (UAV) has been used to assist agricultural production. Precision landing control of UAV is critical for application of it in some specific areas such as greenhouses or livestock/poultry houses. For controlling UAV landing on a fixed or mobile apron/platform accurately, this study proposed an automatic method and tested it under three scenarios: (1) UAV landing at high operating altitude based on the GPS signal of the mobile apron; (2) UAV landing at low operating altitude based on the image recognition on the mobile apron; and (3) UAV landing progress control based on the fixed landing device and image detection to achieve a stable landing action. To verify the effectiveness of the proposed control method, apron at both stationary and mobile (e.g., 3 km/h moving speed) statuses were tested. Besides, a simulation was conducted for the UAV landing on a fixed apron by using a commercial poultry house as a model (135 L × 15 W × 3 H m). Results show that the average landing errors in high altitude and low altitude can be controlled within 6.78 cm and 13.29 cm, respectively. For the poultry house simulation, the landing errors were 6.22 ± 2.59 cm, 6.79 ± 3.26 cm, and 7.14 ± 2.41cm at the running speed of 2 km/h, 3 km/h, and 4 km/h, respectively. This study provides the basis for applying the UAV in agricultural facilities such as poultry or animal houses where requires a stricter landing control than open fields.

Meiotic gatekeeper STRA8 regulates cell cycle by interacting with SETD8 during spermatogenesis.

STRA8 (Stimulated By Retinoic Acid Gene 8) is a retinoic acid (RA) induced gene that plays vital roles in spermatogonial proliferation, differentiation and meiosis. The SETD8 and STRA8 protein interaction was discovered using the yeast two-hybrid technique using a mouse spermatogonial stem cell (SSC) cDNA library. The interaction of these two proteins was confirmed using co-immunoprecipitation and identification of key domains governing the protein: protein complex. STRA8 and SETD8 showed a mutual transcriptional regulation pattern that provided evidence that SETD8 negatively regulated transcriptional activity of the STRA8 promoter. The SETD8 protein directly bound to the proximal promoter of the STRA8 gene. STRA8 increased the transcriptional activity of SETD8 promoter in a dose-dependent manner. For the first time, we have discovered that STRA8 and SETD8 display a cell cycle-dependent expression pattern in germline cells. Expression levels of SETD8 and H4K20me1 in S phase of STRA8 overexpression GC1 cells were different from that previously observed in tumour cell lines. In wild-type mice testis, SETD8, H4K20me1 and PCNA co-localized with STRA8 in spermatogonia. Further, our studies quantitated abnormal expression levels of cell cycle and ubiquitination-related factors in STRA8 dynamic models. STRA8 and SETD8 may regulate spermatogenesis via Cdl4-Clu4A-Ddb1 ubiquitinated degradation axis in a PCNA-dependent manner.

Stra8 may inhibit apoptosis during mouse spermatogenesis via the AKT signaling pathway.

Stimulated by retinoic acid 8 (Stra8), one of genes induced by retinoic acid (RA), is required for the meiotic initiation of male spermatogenesis. The present study found that Stra8 inhibited apoptosis in male Stra8­knockout mice, and in mice with vitamin A deficiency and vitamin A recovery in vivo. This phenotype was also verified in GC1 spermatogonia (spg) cells overexpressing Stra8. In addition, microarray analysis identified that there were nine differentially expressed genes (DEGs) in the Stra8­overexpressed GC1 spg cells compared with the control groups; the expression of these nine genes was verified via mRNA expression levels. The DEGs were as follows: Phosphatidylinositol­dependent kinase 1 (PDK1), a key gene upstream of protein kinase B (AKT); angiopoietin 2, a B­cell lymphoma 2 (Bcl­2)­inhibited gene; transcription factor 4, glutathione S­transferase P91 and ubiquitin­specific protease 33, mitogen­activated protein kinase (MAPK)­related genes; oxidative stress induced growth inhibitor 1, related to the P53 pathway; Bcl­2, P53, ERK (MAPK1/3), c­Jun N­terminal kinase (MAPK8/9), and P38 (MAPK14), all of which are key genes involved in the AKT signaling pathway. Therefore, the present study further verified these genes and found that the mRNA and protein expression levels of PDK1, AKT, Bcl­2 and ERK were increased. Although the mRNA expression level of P53 was decreased, there was no significant difference in the protein expression level in Stra8­overexpressing GC1 spg cells compared with controls. In addition, Caspase 3, one of the executioner caspases, was decreased in Stra8­overexpressing GC1 spg cells compared with the control groups. Therefore, it was suggested that Stra8 may directly or indirectly inhibit caspases through the AKT signaling pathway and ultimately exert an anti­apoptotic effect in the male reproductive system.

[Preparation and application of rabbit anti-mouse Setd8 polyclonal antibody].

Objective: To purify the recombinant Setd8 protein and prepare rabbit anti-mouse Setd8 polyclonal antibody. Methods: The recombinant plasmid p ET-30a-Setd8 was constructed by double enzyme digestion and linkage,and then transformed into E. coli BL21. The expression of the target protein was induced by IPTG and the expression product was purified by Ni-NTA affinity chromatograph. The purified protein was used to immunize New Zealand white rabbits to produce polyclonal antibody. The titer and specificity of the antibody were identified by ELISA,Western blotting and immunohistochemistry. Results: The prokaryotic expression vector p ET-30a-Setd8 was constructed successfully. After induced by IPTG,the recombinant Setd8 protein was expressed effectively in E. coli BL21. Polyclonal antibody against Setd8 was generated by immunizing rabbits with the routine method. ELISA showed that the titer of rabbit anti-Setd8 antiserum was 1 ∶ 1 000 000.Western blotting demonstrated that the polyclonal antibody could recognize the native mouse Setd8 protein. Immunohistochemistry revealed that Setd8 protein recognized by the polyclonal antibody was mainly distributed in the nucleus of spermatogonia in adult mouse testis. Conclusion: Using the prokaryotic expression vector p ET-30a-Setd8,we have prepared successfully the polyclonal antibody with high affinity and specificity.

[Sperm acrosome formation-associated genes in mice: Advances in studies].

Spermiogenesis is a complex process of differentiation and morphologic alteration, in which sperm acrosome formation is an important stage. Acrosome is an essential component of the sperm head, which develops in four distinct phases: Golgi, cap, acro- somal, and maturation, each supported by precise and orderly regulation of various genes. The regulatory genes which act on Golgi ap- paratus include GOPC, Hrb, SPATA16, PICK1, and CK2α', those involved in the cap phase are Fads2, syntaxin 2, Kdm3a, and UBR7, and participating in acrosomal and maturation phases are KIFC1, Rnf19a, and DPY19L2. The abnormalities of these genes may affect male fertility by influencing the connection of the nuclear dense lamina and acroplaxome with the nuclear membrane and then the fusion and transportation of vesicles. This review focuses on the genes involved in different phases of acrosome formation.

Ubiquitously expressed transcript is a novel interacting protein of protein inhibitor of activated signal transducer and activator of transcription 2.

Protein inhibitor of activated signal transducer and activator of transcription 2 (PIAS2) is a member of the PIAS protein family. This protein family modulates the activity of several transcription factors and acts as an E3 ubiquitin ligase in the sumoylation pathway. To improve understanding of the physiological roles of PIAS2, the current study used a yeast two­hybrid system to screen mouse stem cell cDNA libraries for proteins that interact with PIAS2. The screening identified an interaction between PIAS2 and ubiquitously expressed transcript (UXT). UXT, also termed androgen receptor trapped clone­27, is an α­class prefoldin­type chaperone that acts as a coregulator for various transcription factors, including nuclear factor­κB and androgen receptor (AR). A direct interaction between PIAS2 and UXT was confirmed by direct yeast two­hybrid analysis. In vitro evidence of the association of UXT with PIAS2 was obtained by co­immunoprecipitation. Colocalization between PIAS2 and UXT was identified in the nucleus and cytoplasm of HEK 293T and human cervical carcinoma HeLa cells. The results of the current study suggested that UXT is a binding protein of PIAS2, and interaction between PIAS2 and UXT may be important for the transcriptional activation of AR.