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This file contains 9 supplemental figures and 3 supplemental tables. Supplemental Figure 1. There are four potential binding sites (red) for NF-Y in the 1kb upstream region of the ZDHHC5 promoter. Supplemental Figure 2. The mutant p53 and NF-Y co-recruit onto the ZDHHC5 promoter. Supplemental Figure 3. Genetic, differentiated and proliferative characteristics of CA-KRAS/hTERT/DN-p53/ZDHHC5 hNSCs. Supplemental Figure 4. There is co-expression of ZDHHC5 and SOX2 or CD133 in p53 mutant glioma samples and glioma stem cells. Supplemental Figure 5. ZDHHC5 regulates self-renewal of p53 mutant GSCs. Supplemental Figure 6. ZDHHC5 regulates tumor angiogenesis in xenografted brain tumors from p53 mutant GSCs. Supplemental Figure 7. ZDHHC5 specifically regulates EZH2 palmitoyaltion level and EZH2-mediated colony formation and cell migartion. Supplemental Figure 8. The EZH2 potential palmitoyaltion sites which are conserved in different species. Supplemental Figure 9. ZDHHC5 regulates EZH2-mediated glioma development. Supplemental Table1. p53 mutational status in glioma. Supplemental Table2. The Relationship between ZDHHC5 expression and p53 mutantional status in glioma. Supplemental Table 3. Limiting dilution tumor formation of p53 mutant GSCs transfected with or without ZDHHC5 shRNA.

Gliomas with mutant p53 occurring in 30% of glioma patients exhibit therapeutic resistance and poor outcomes. In this study, we identify a novel mechanism through which mutant p53 drives cancer cell survival and malignant growth. We documented overexpression of the zinc finger protein ZDHHC5 in glioma compared with normal brain tissue and that this event tightly correlated with p53 mutations. Mechanistic investigations revealed that mutant p53 transcriptionally upregulated ZDHHC5 along with the nuclear transcription factor NF-Y. These events contributed to the development of glioma by promoting the self-renewal capacity and tumorigenicity of glioma stem-like cells, by altering the palmitoylation and phosphorylation status of the tumor suppressor EZH2. Taken together, our work highlighted ZDHHC5 as a candidate therapeutic target for management of p53-mutated gliomas. Cancer Res; 77(18); 4998–5010. ©2017 AACR.

Based on Timoshenko beam theory and considering the layer bending deformation and shear deformation of the structure, a semi analytical solution method (SAM) is proposed for the interlayer isolation structure system. The equivalent mechanical model of cantilever beam is established by using the distributed parameter system, on which the dynamic partial differential equation is derived; the boundary condition and continuity condition are used to solve the equation to obtain the natural vibration characteristics; the orthogonal conditions are deduced by Betti’s law and the seismic response of the structure can be solved by the mode superposition method. A 10 story isolated structure is designed as well as analysed by the semi analytical method (SAM) and the ETABS finite element simulation (FESE) separately. The results show that: the existence of the isolation layer may reduce the response of the superstructure and amplify the response of the substructure, and the amplification can be solved by increasing the mass, bending stiffness of the superstructure and shear stiffness of the isolation layer. In regular interlayer isolation structure, the optimal frequency ratio of the superstructure and substructure increases with the rising of isolation layer position. The results also prove the correctness and accuracy of the SAM, which enriches the analysis theory of interlayer isolation structure and provides reference for the accurate analysis of structural response in engineering.

A cantilever beam with multiple rotation spring-dampers was utilized to simulate the damped outrigger system. Two new methods, namely, the “Maxwell dampers type method” and the “virtual small mass method,” were proposed to present the simplified seismic response analysis of the damped outrigger system. Corresponding dynamic characteristic equations based on the proposed methods were also given. Subsequently, [Formula: see text] performance index was employed to obtain the optimal damper parameters. The numerical analysis shows that the proposed methods agree well with the finite-element method and have relatively accurate results. Stiffness ratios of the perimeter columns have a significant effect on the pseudo frequencies and system damping ratios. Lower stiffness ratios of the perimeter columns commonly lead to higher pseudo frequencies and system damping ratios. Furthermore, the damped outrigger system has excellent seismic mitigation performance when applied with appropriate damping parameters based on [Formula: see text].

The target of EGR1 protein 1 (TOE1) is evolutionarily conserved from Caenorhabditis elegans to mammals, which plays a critical role in the maturation of a variety of small nuclear RNAs. Mutation in human TOE1 has been reported to cause pontocerebellar hypoplasia type 7, a severe neurodegenerative syndrome. However, the role of TOE1 in early embryonic development remains unclear. Herein, we found that Toe1 mRNA and protein were expressed in mouse preimplantation embryos. Silencing Toe1 by siRNA led to morula-to-blastocyst transition failure. This developmental arrest can be rescued by Toe1 mRNA microinjection. EdU incorporation assay showed a defect in blastomere proliferation within developmentally arrested embryos. Further studies revealed that Toe1 knockdown caused increased signals for γH2AX and micronuclei, indicative of sustained DNA damage. Moreover, mRNA levels of cell cycle inhibitor p21 were significantly upregulated in Toe1 knockdown embryos before developmental arrest. Together, these results suggest that TOE1 is indispensable for mouse early embryo development potentially through maintaining genomic integrity. Our findings provide further insight into the role of TOE1 in mouse preimplantation embryonic development.

Considering the uncertainties of seismic excitation and stochastic parameters of the Inter-story Isolation Structure, the implicit structural response function is transformed into an explicit one by the Stochastic Response Surface Method (SRSM), combined with a probabilistic collocation method based on the linear independence principle. By using the reliability analysis method based on the first-passage failure, the reliability of isolation structure is analyzed and the sensitivity of the reliability of each sub-structure is further analyzed. The results show that conducting sensitivity research based on reliability analysis can provide an important theoretical basis for an optimized and robust design of a random vibration system. The changes of the stiffness and damping ratio of the Isolation Layer have a great influence on the reliability of the system and have different effects on the reliability of different parts of the system. The analysis results of reliability sensitivity could provide system reliability indexes to rank stochastic parameters by their importance, thus improving the efficiency of the structure reliability analysis and optimization design.

The Compositive Passive Control method (CPC method) of “interlayer seismic isolation” + “shock absorption between adjacent towers” is applied to Multi-Tower Building (MTB) with a large podium. Taking the total vibration energy of the structure as the optimal control objective, the response expressions of a multi-DOF layer shear model and an equivalent single-DOF layer shear model are derived. The rationality of the equivalent model is demonstrated from the perspective of mode characteristics and time history response. Based on the Kanai–Tajimi spectral seismic motion model, the control effect and the optimal control parameters of the CPC method are studied. Compared with the conventional seismic scheme and the interlayer seismic isolation scheme, the influence of the CPC method on the structure natural vibration characteristics and dynamic response under different parameters of the connecting control device is discussed and analyzed. The numerical analysis proves that the CPC method provides a significant damping effect compared with the interlayer seismic isolation scheme. It maintains the advantages of the interlayer seismic isolation scheme and reduces the defect of the response amplification caused by the interlayer isolation. This analysis constitutes a good reference for the follow-up study of the CPC method of MTB.

To reveal the effects of repetitive transcranial magnetic stimulation (rTMS) on the improvement of cognitive function in patients with stress-related depression, and to enrich the neural mechanism(s) underlying rTMS so as to improve cognitive function in patients with stress-related depression.We conducted a randomized, double-blind, placebo-controlled study of rTMS in patients with stress-related depression who were 18-40 years of age. Patients were randomly allocated to either a sham or experimental group in a 1:1 ratio. A 10-session rTMS protocol was used with 10-Hz stimulation over the left dorsolateral prefrontal cortex (DLPFC). Clinical assessments (HAMD, HAMA, DASS, MoCA), neuropsychologic (Stroop, WCST), and resting state fMRI andrTMS relieved the mental symptoms of patients in both groups. The MoCA score of patients in the experimental group increased; the number of correct answers increased significantly in Stroop testing, and the number of errors and omissions decreased significantly; the number of persistent errors decreased significantly; and the time used to complete the test decreased to an even greater extent in the WCST experimental group. The ReHo value in the lingual gyrus of the right hemisphere and the cuneus of the left and right hemispheres in the experimental group decreased after treatment. The DC value in the left and right hemispheric cuneus and postcentral gyrus of the left hemisphere in the experimental group diminished after treatment. The functional connections of these brain regions also changed as the Cho and NAA/Cr of the left DLPFC changed, with alterations related to the improvement in cognitive function. The level of choline (Cho) in the left DLPFC of the experimental group was significantly lower than that of the control group, and the level of N-acetylaspartate/creatine (NAA/Cr) in the left DLPFC of the control group was significantly higher than that of the experimental group. These changes were related to the overall improvement in cognitive function.Ten-Hz rTMS over the left DLPFC improved the cognitive function of patients with stress-related depression. The governing mechanism for this phenomenon may be via rTMS effects on multiple visual-related brain regions and their functional connections, and on the somatosensory cortex and its functional connection with visual and auditory cortex, reducing the level of Cho and stabilizing the level of NAA/Cr in the left DLPFC.

In this study, we sought to investigate the association between N6-methyladenosine (mFirst, the clinical data and the transcriptomes of 530 patients with ccRCC were downloaded from The Cancer Genome Atlas (TCGA). The expression patterns of mAmong the 13 mFollowing TCGA data-mining, different molecular subtypes of ccRCC based on m

Gliomas with mutant p53 occurring in 30% of glioma patients exhibit therapeutic resistance and poor outcomes. In this study, we identify a novel mechanism through which mutant p53 drives cancer cell survival and malignant growth. We documented overexpression of the zinc finger protein ZDHHC5 in glioma compared with normal brain tissue and that this event tightly correlated with p53 mutations. Mechanistic investigations revealed that mutant p53 transcriptionally upregulated ZDHHC5 along with the nuclear transcription factor NF-Y. These events contributed to the development of glioma by promoting the self-renewal capacity and tumorigenicity of glioma stem-like cells, by altering the palmitoylation and phosphorylation status of the tumor suppressor EZH2. Taken together, our work highlighted ZDHHC5 as a candidate therapeutic target for management of p53-mutated gliomas. Cancer Res; 77(18); 4998–5010. ©2017 AACR.

Supervillin is an actin-associated protein that regulates actin dynamics by interacting with Myosin II, F-actin, and Cortactin to promote cell contractility and cell motility. Two splicing variants of human Supervillin (SV1 and SV4) have been reported in non-muscle cells; SV1 lacks 3 exons present in the larger isoform SV4. SV2, also called archvillin, is present in striated muscle; SV3, also called smooth muscle archvillin or SmAV, was cloned from smooth muscle. In the present study, we identify a novel splicing variant of Supervillin (SV5). SV5 contains a new splicing pattern. In the mouse tissues and cell lines examined, SV5 was predominantly expressed in skeletal and cardiac muscles and in proliferating cells, but was virtually undetectable in most normal tissues. Using RNAi and rescue experiments, we show here that SV5 displays altered functional properties in cancer cells, and regulates cell proliferation and cell migration.

Uneven foundation settlement is one of the common engineering problems in a collapsible loess area. In order to study the influence of uneven foundation settlement on the seismic performance of a structure, the incremental dynamic analysis (IDA) method is used to analyze the seismic vulnerability of the steel structure frame. The differences in the seismic response of the structure in relation to uneven foundation settlement are analyzed. The influences of uneven foundation settlement quantities and various areas of uneven settlement on the seismic response of the structure are discussed. On this basis, the relationship between ground motion intensity and structural failure probability is studied, by changing the magnitude of seismic acceleration peaks. Compared with the unsettled structure, the internal force redistribution of the structure caused by uneven foundation settlement is one of the causes of earthquake damage for some components. The uneven foundation settlement located at the corner of the plane of the structure is likely to cause more serious earthquake damage to the structure. Uneven settlement will cause an increase in storey drift. With the increase in settlement, the seismic damage of the superstructure will be aggravated.

The voltage-gated proton channel Hv1 is a potent acid extruder that participates in the extrusion of the intracellular acid. Here, we showed for the first time, Hv1 is highly expressed in mouse and human pancreatic islet β-cells, as well as β-cell lines. Imaging studies demonstrated that Hv1 resides in insulin-containing granules in β-cells. Knockdown of Hv1 with RNA interference significantly reduces glucose- and K(+)-induced insulin secretion in isolated islets and INS-1 (832/13) β-cells and has an impairment on glucose- and K(+)-induced intracellular Ca(2+) homeostasis. Our data demonstrated that the expression of Hv1 in pancreatic islet β-cells regulates insulin secretion through regulating Ca(2+) homeostasis.

A multifactor coupling test protocol was designed to determine the freeze–thaw resistance of concrete with fly ash as fine aggregate (CFA). Rapid freeze–thaw tests were conducted to measure the mass loss and relative dynamic modulus of elasticity of concrete specimens to evaluate their freeze–thaw resistance. The test results show that the freeze–thaw resistance of the concrete decreases with the increase in the initial stress damage. The coupling effect of stress damage and high temperature accelerates the attenuation of mass and relative dynamic modulus of elasticity of concrete. CFA has a higher freeze–thaw resistance than concrete.

Background Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world and metastasis is the leading cause of death associated with HCC. Hypoxia triggers the epithelial-mesenchymal transition (EMT) of cancer cells, which enhances their malignant character and elevates metastatic risk. Supervillin associates tightly with the membrane and cytoskeleton, promoting cell motility, invasiveness, and cell survival. However, the roles of supervillin in HCC metastasis remain unclear. Methods Tissue microarray technology was used to immunohistochemically stain for supervillin antibody in 173 HCC tissue specimens and expression levels correlated with the clinicopathological variables. Tumor cell motility and invasiveness, as well as changes in the mRNA expression levels of genes associated with cancer cell EMT, were investigated. The relationship between supervillin and Rho GTPases was examined using Co-IP and GST pull-down. Results Hypoxia-induced upregulation of supervillin promoted cancer cell migration and invasion via the activation of the ERK/p38 pathway downstream of RhoA/ROCK signaling. Furthermore, supervillin regulated the expression of EMT genes during hypoxia and accelerated the metastasis of HCC in vivo. Conclusions Hypoxia-induced increase in supervillin expression is a significant and independent predictor of cancer metastasis, which leads to poor survival in HCC patients. Our results suggest that supervillin may be a candidate prognostic factor for HCC and a valuable target for therapy. Electronic supplementary material The online version of this article (10.1186/s13046-018-0787-2) contains supplementary material, which is available to authorized users.

More than 40% of glioma patients have tumors that harbor PTEN (phosphatase and tensin homologue deleted on chromosome ten) mutations; this disease is associated with poor therapeutic resistance and outcome. Such mutations are linked to increased cell survival and growth, decreased apoptosis, and drug resistance; thus, new therapeutic strategies focusing on inhibiting glioma tumorigenesis and progression are urgently needed. Melatonin, an indolamine produced and secreted predominantly by the pineal gland, mediates a variety of physiological functions and possesses antioxidant and antitumor properties. Here, we analyzed the relationship between PTEN and the inhibitory effect of melatonin in primary human glioma cells and cultured glioma cell lines. The results showed that melatonin can inhibit glioma cell growth both in culture and in vivo. This inhibition was associated with PTEN levels, which significantly correlated with the expression level of MT1 in patients. In fact, c-fos-mediated MT1 was shown to be a key modulator of the effect of melatonin on gliomas that harbor wild type PTEN. Taken together, these data suggest that melatonin-MT1 receptor complexes represent a potential target for the treatment of glioma.

Table S1. Stealth siRNA used in the study. Figure S1. siRNA that targeted supervillin splice isoforms (SV1, SV4, and SV5) reduced the level of each isoform in HCC cells, and the up-regulation of supervillin was not dependent on HIF1Îą-induced transcriptional activation. Figure S2. Supervillin promotes HCC migration and invasion during hypoxia. Figure S3. Quantification of EMT markersâ expression levels regulated by supervillin during hypoxia. Figure S4. Supervillin regulated HCC cell migration and invasion through the RhoA/ROCK pathway during hypoxia. Figure S5. Supervillin regulates RhoA activation and ERK/p38 transduction during hypoxia. Figure S6. Quantification of phosphorylated p38 and ERK levels regulated by supervillin-RhoA/ROCK signaling during hypoxia. Figure S7. Supervillin-RhoA/ROCK-ERK/p38 signaling pathway in HCC metastasis under hypoxia in vivo. Figure S8. Supervillin promotes Rac1 activity and Rac-mediated cell metastasis in normoxia and hypoxia. Figure S9. RhoA activation and MAPK signaling vary in cell types and environments. Figure S10. There was not obvious effect on microtubule dynamics upon supervillin knockdown. Figure S11. The inhibitors used in this study did not affect cell apoptosis and viability. (PDF 2231 kb)

// Xueran Chen 1, 2 , Zhen Wang 1, 2 , Huihui Ma 3, 4 , Shangrong Zhang 1, 2 , Haoran Yang 1, 2 , Hongzhi Wang 1, 2 and Zhiyou Fang 1, 2 1 Cancer Hospital, Chinese Academy of Sciences, Hefei, Anhui, 230031, China 2 Anhui Province Key Laboratory of Medical Physics and Technology, Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, China 3 Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui, 230031, China 4 Department of Radiation Oncology, First Affiliated Hospital, Anhui Medical University, Hefei, Anhui, 230022, China Correspondence to: Xueran Chen, email: xueranchen@cmpt.ac.cn Zhiyou Fang, email: z.fang@cmpt.ac.cn Keywords: melatonin, CCL20, epithelial–mesenchymal transition (EMT), glioma Received: July 17, 2017 Accepted: August 01, 2017 Published: August 24, 2017 ABSTRACT Tumor recurrence in gliomas is partly attributed to increased epithelial–mesenchymal transition (EMT) and enhanced tumor cell dissemination in the adjacent brain parenchyma. Thus, exploring effective strategies for against EMT-like changes in glioma invasion and recurrence will be important for glioma treatment. In this study, we investigated the roles of melatonin in hypoxia-induced EMT suppression, and found that melatonin could significantly suppress the release of the cytokine, CCL20, from cancer cells and antagonize glioma cell metastasis and invasion under hypoxic stress in glioma cells. Furthermore, our findings show that melatonin deregulates Smad7 expression to suppress TGFβ/Smad-mediated increase in CCL20 transcript levels and CCL20-induced EMT occurrence, suggesting a potential anti-EMT therapeutic role for melatonin in malignant transformation in gliomas.

Here, we demonstrate that the voltage-gated proton channel Hv1 represents a regulatory mechanism for insulin secretion of pancreatic isletβcell.In vivo, Hv1-deficient mice display hyperglycemia and glucose intolerance due to reduced insulin secretion, but normal peripheral insulin sensitivity.In vitro, islets of Hv1-deficient and heterozygous mice, INS-1 (832/13) cells with siRNA-mediated knockdown of Hv1 exhibit a marked defect in glucose- and K+-induced insulin secretion. Hv1 deficiency decreases both insulin and proinsulin contents, and limits glucose-induced Ca2+entry and membrane depolarization. Furthermore, loss of Hv1 increases insulin-containing granular pH and decreases cytosolic pH. In addition, histologic studies show a decrease inβcell mass in islets of Hv1-deficient mice. Collectively, our results indicate that Hv1 supports insulin secretion in theβcell by calcium entry, membrane depolarization and intracellular pH regulation.SIGNIFICANCE STATEMENTThe voltage-gated proton channel Hv1 is highly expressed in insulin-containing granules in pancreatic β cells. Hv1 supports insulin secretion in theβcell by calcium entry, membrane depolarization and regulation of intragranular and cytosolic pH, which represents a regulatory mechanism for insulin secretion of pancreatic isletβcell. Our research demonstrates that Hv1 expressed inβcell is required for insulin secretion and maintains glucose homeostasis, and reveals a significant role for the proton channel in the modulation of pancreaticβcell function.

The increased glycolysis and proton secretion in tumors is proposed to contribute to the proliferation and invasion of cancer cells during the process of tumorigenesis and metastasis. Here, treatment of human breast cancer cells with proton pump inhibitor (PPI) lansoprazole (LPZ) induces cell apoptosis in a dose-dependent manner. In the implantation of the MDA-MB-231 xenografts in nude mice, administration of LPZ significantly inhibits tumorigenesis and induces large-scale apopotosis of tumor cells. LPZ markedly inhibits intracellular proton extrusion, induces an increase in intracellular ATP level, lysosomal alkalinization and accumulation of reactive oxygen species (ROS) in breast cancer cells. The ROS scavenger N-acetyl-l-cysteine (NAC) and diphenyleneiodonium (DPI), a specific pharmacological inhibitor of NADPH oxidases (NOX), significantly abolish LPZ-induced ROS accumulation in breast cancer cells. Our results suggested that LPZ may be used as a new therapeutic drug for breast tumor.

Highlights: •Hv1 is expressed in highly metastatic glioma cell. •Zn{sup 2+} ions induces apoptosis in highly metastatic glioma cells. •Zn{sup 2+} ions markedly inhibit proton secretion. •Zn{sup 2+} ions reduce the gelatinase activity. •Inhibition of Hv1 activity via Zn{sup 2+} ions can effectively retard the cancer growth. -- Abstract: In contrast to the voltage-gated K{sup +} channels, the voltage-gated proton channel Hv1 contains a voltage-sensor domain but lacks a pore domain. Here, we showed that Hv1 is expressed in the highly metastatic glioma cell SHG-44, but lowly in the poorly metastatic glioma cell U-251. Inhibition of Hv1 activity by 140 μM zinc chloride induces apoptosis in the human highly metastatic glioma cells. Zn{sup 2+} ions markedly inhibit proton secretion, and reduce the gelatinase activity in the highly metastatic glioma cells. In vivo, the glioma tumor sizes of the implantation of the SHG-44 xenografts in nude mice that were injected zinc chloride solution, were dramatically smaller than that in the controlled groups. The results demonstrated that the inhibition of Hv1 activity via Zn{sup 2+} ions can effectively retard the cancer growth and suppress the cancer metastasis by the decrease of proton extrusion and the down-regulation of gelatinase activity. Our results suggestmore » that Zn{sup 2+} ions may be used as a potential anti-glioma drug for glioma therapy.« less

Solid tumors exist in a hypoxic microenvironment, and possess high-glycolytic metabolites. To avoid the acidosis, tumor cells must exhibit a dynamic cytosolic pH regulation mechanism(s). The voltage-gated proton channel Hv1 mediates NADPH oxidase function by compensating cellular loss of electrons with protons. Here, we showed for the first time, that Hv1 expression is increased in colorectal tumor tissues and cell lines, associated with poor prognosis. Immunohistochemistry showed that Hv1 is strongly expressed in adenocarcinomas but not or lowly expressed in normal colorectal or hyperplastic polyps. Hv1 expression in colorectal cancer is significantly associated with the tumor size, tumor classification, lymph node status, clinical stage and p53 status. High Hv1 expression is associated significantly with shorter overall and recurrence-free survival. Furthermore, real-time RT-PCR and immunocytochemistry showed that Hv1 is highly expressed in colorectal cancer cell lines, SW620, HT29, LS174T and Colo205, but not in SW480. Inhibitions of Hv1 expression and activity in the highly metastatic SW620 cells by small interfering RNA (siRNA) and Zn(2+) respectively, markedly decrease the cell invasion and migration, restraint proton extrusion and the intracellular pH recovery. Our results suggest that Hv1 may be used as a potential biomarker for diagnosis and prognosis of colorectal carcinoma, and a potential target for anticancer drugs in colorectal cancer therapy.