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This paper focuses on coupling property and dynamic stability of spinning missiles equipped with a two-loop autopilot. The closed-loop characteristic equation is established in the complex form, on which we investigated the dominant factors of the cross coupling, i.e., actuator dynamics, airframe configuration and autopilot, to provide physical understandings of coupling property. By coupling analysis, it is found that coupling can be mitigated by the autopilot. However, the proposed static decoupling approach is necessary to eliminate steady coupling completely. Furthermore, by solving the forth-order characteristic equations with Cardan formula, the dynamic stability boundary is derived numerically. As actuator dynamics is extremely significant to stability, the proposed stability boundary is derived with consideration of it, and thus, it is more practical than results in previous studies. The effectiveness of decoupling method and the correctness of dynamic stability boundary is verified by mathematical simulations. Theoretical and numerical results also reveal that high actuator bandwidth and less time delay is beneficial to dynamic stability.

Spatio-temporal dynamics and stakeholder engagement are critical components in the assessment of ecosystem services to support well-targeted and localised strategies for sustainable ecosystem management. Here, we investigated the recent changing pattern of ecosystem services’ value (ESV) in the Yangtze River Middle Reaches Megalopolis (YRMRM) - a region of vital economic and ecological importance in central China. Local experts’ opinions on the regional importance of different ecosystem services were acquired from an extensive survey and integrated into the localisation of ESV. We explored the shifting extents of the hot and cold spots of ESV and whether the ESV of a place could be related to its geographical location and distance. Our results showed a downtrend of ESV from 2000 to 2015, resulting from a loss in cultivated land and thereby a reduced ESV of the food supply. Cold spots intensified while hot spots diminished across the YRMRM. The ESVs of a place were distance-decay functions of the distances (d) between the place to the nearest provincial centre, lake and the Yangtze River. The maximum ESV was usually located at 42 km away from nearest provincial centre, 10 km away from the Yangtze River, and near the lake. This study adds important first-hand empirical data to the field of the stakeholder-driven valuation of localised ESV. Our results based on a comprehensive spatiotemporal assessment of ESV is conducive to promoting the coordination of sustainable development and ecological protection in YRMRM.

Products (such as agricultural products and handicrafts) that are assisted by enterprises in their production or sales and whose sales revenues are dedicated to anti-poverty campaigns in a particular area are regarded as poverty alleviation products (PAPs). This study focuses on PAP advertisements and analyses the interaction between the expression of different emotions (positive expression/negative expression) by models in PAP advertisements and the types of products (general products/PAPs). This study also examines the expression framework of these advertisements (gain-framed/loss-framed) and consumer purchase intentions. The results of Study 1 indicate that consumers demonstrate a preference for PAPs and that negative expressions significantly improve consumers' purchase preference for PAPs. Study 2 demonstrates that this effect results from the completely mediating effect of consumer guilt. However, in Study 3, when an advertisement employs a combination of a “loss-framed message and negative facial expression”, it fails to further improve consumer purchase intention compared to other groups. The conclusions of this examination not only support the enterprises that positively participate in industrial poverty alleviation but also provide theoretical guidance to enterprises in the design of PAP advertisements.

This paper presents a novel guidance law considering the seeker dynamics for manoeuvring targets to achieve short homing time guidance using the Lyapunov method. Based on linear and nonlinear kinematics, a Lyapunov-based guidance law is synthesised to compensate for the seeker’s first-order lag. The closed-form solution of the proposed guidance system is also derived analytically. To implement the proposed guidance law, a Kalman filter algorithm is presented to extract the line-of-sight rate and its higher order derivative. Numerical simulations are carried out to demonstrate the effectiveness of the proposed guidance law under various conditions. Monte Carlo simulations are also performed to test the robustness against measurement noise.

// Rongfei Han 1, * , Guanqun Huang 1, * , Yejun Wang 1 , Yafei Xu 1 , Yueming Hu 1 , Wenqi Jiang 1 , Tianfu Wang 2 , Tian Xiao 1 , Duo Zheng 1 1 Shenzhen Key Laboratory of Translational Medicine of Tumor, Department of Cell Biology and Genetics, Shenzhen University Health Sciences Center, Shenzhen, Guangdong, 518060, P.R.China 2 Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, School of Biomedical Engineering, Shenzhen University Health Sciences Center, Shenzhen, Guangdong, 518060, P.R.China * These authors contributed equally to this work Correspondence to: Tianfu Wang, email: tfwang@szu.edu.cn Tian Xiao, email: txiao@szu.edu.cn Duo Zheng, email: dzheng@szu.edu.cn Keywords: gene expression noise, p53, immune activity, cancer prognosis Received: July 26, 2016 Accepted: September 29, 2016 Published: October 04, 2016 ABSTRACT Gene expression in metazoans is delicately organized. As genetic information transmits from DNA to RNA and protein, expression noise is inevitably generated. Recent studies begin to unveil the mechanisms of gene expression noise control, but the changes of gene expression precision in pathologic conditions like cancers are unknown. Here we analyzed the transcriptomic data of human breast, liver, lung and colon cancers, and found that the expression noise of more than 74.9% genes was increased in cancer tissues as compared to adjacent normal tissues. This suggested that gene expression precision controlling collapsed during cancer development. A set of 269 genes with noise increased more than 2-fold were identified across different cancer types. These genes were involved in cell adhesion, catalytic and metabolic functions, implying the vulnerability of deregulation of these processes in cancers. We also observed a tendency of increased expression noise in patients with low p53 and immune activity in breast, liver and lung caners but not in colon cancers, which indicated the contributions of p53 signaling and host immune surveillance to gene expression noise in cancers. Moreover, more than 53.7% genes had increased noise in patients with late stage than early stage cancers, suggesting that gene expression precision was associated with cancer outcome. Together, these results provided genomic scale explorations of gene expression noise control in human cancers.

The bactericidal/permeability-increasing protein (BPI) with bactericidal and endotoxin-neutralizing activity is of considerable interest in clinical applications. However, the crucial residues responsible for the bactericidal activity of BPI remain elusive. In previous study, we identified the mutation of mBPI5 associated with the male infertility of mice. Here, the effects of Asp190Ala mutation on the antibacterial activity of mBPI5 have been determined. Substitution of Asp190 by alanine caused significant improvement in cytotoxic effect toward both E.coli J5 and P.aeruginosa. Liposome co-sedimentation assay showed that the ratio of Asp190Ala mutant binding to lipids increased by 8 folds. These results were well consistent with known fact that antibacterial activity of BPI is attributed to its high affinity for lipid moiety of lipopolysaccharides (LPS). The constructed structure of mBPI5 revealed that Asp190 was located close to 6 positively charged residues on the surface of N-terminal domain. When replacing Asp190 with alanine, salt linkages with Arg188 were broken, making the side chain of Arg188 be free to move and form tighter contacts with negatively charged LPS. These findings suggest that residue 190 combined with surrounding positively charged residues largely contribute to bactericidal and endotoxin-neutralizing activities of mBPI5.

In this paper, a new optimal linear-quadratic guidance law with terminal acceleration constraints is proposed for intercepting maneuvering target. Because the angle of attack is approximately proportional to the normal acceleration for aerodynamic control missile, not only miss distance, but also terminal acceleration is required to converge to zero for increasing effectiveness of the warhead for destroying armored target in the proposed guidance law. The gain characteristic of proposed guidance law is studied by comparing with traditional optimal guidance law with considering autopilot dynamics lags. Numerical simulations are performed to examine performance of the proposed optimal guidance law, and its robustness with respect to the time constant of autopilot dynamic lags is also studied by considering time constant typical errors.

While planning and management of urban green space (UGS) remain primarily driven by policymakers and expert knowledge, increasing attention has been paid to the general public's perception of and demand for UGS. This study was conducted to provide price-related outcomes to support UGS-related decision making and achieve an improved understanding of the causal relationships underpinning the residents' willingness-to-pay (WTP) for UGS conservation. An extensive survey with 3000 urban residents from three provincial capital cities in central China (i.e., Wuhan, Changsha and Nanchang) was conducted. We applied structural equation modelling to examine a range of hypothetical causal relationships among residents' perceptions about ecosystem services/disservices provided by UGS, frequency of UGS visits, socioeconomic status (SES) and WTP for the conservation of five different types of UGS. We found that residents had an average WTP of 202.4 CNY or 30.6 USD per year cross UGS types, with the attached green space having the greatest WTP (223.0 CNY or 33.7 USD per year), followed by park (215.4 CNY or 32.6 USD year), square (201.7 CNY or 30.5 USD year), suburban ecological (190.1 CNY or 28.7 USD year) and protective (182.0 CNY or 27.5 USD year) green spaces. Perceptions of UGS's ecosystem services had a positive causal impact on WTP, while perceptions of ecosystem disservices would have a negative impact. The frequency of UGS visits was found to amplify WTP at a limited level. It could further heighten and diminish the perceptions of ecosystem services and disservices. Participants with higher SES typically had greater perceptions of UGS's ecosystem services and WTP. Finally, we found that the structure of the causal relationships on WTP varied between cities, highlighting the importance of considering the fine-level impacts of biophysical environment jointly. Our results could help urban planners to understand better the ways urban green space was perceived and anticipate the likely effects of changing spatial patterns of UGS on the benefits and nuisances experienced by the general public.

China has experienced rapid residential land expansion in both urban and rural areas over the past three decades, causing complex ecological and environmental challenges. Much research attention has been paid on urbanisation, yet little is known about rural development. In this study, we analysed and compared the changes in a selected number of landscape indices describing the spatial patterns of both urban and rural area in the Middle Reaches of the Yangtze River in central China from 2005 to 2015 and explored how these changes could be associated with the development of high-speed rail (HSR) using spatial error models. We found a partial synchronised spatial development pattern between urban and rural areas in central China, with an increasingly fragmented pattern for both urban and rural areas, albeit rural areas were expanded in a less contiguous but more complex and dispersed fashion. The impacts of the provision of HSR services on the region's spatial development were found to be multi-level. It was associated with greater urban expansion and dispersion at the county/district level and amplified rural patch size and complexity at the patch level. The departure frequency of HSR trains and proximity to HSR station were found to have affected the magnitude of the impact of HSR service provision on regional spatial development. Our results shed lights on the spatio-temporal evolution of an ecologically important region, add new evidence into the expanding fields of urban and rural morphological studies in China, and provide valuable decision support information for integrated spatial planning of transportation and land use.

Deubiquitinase BRISC complex plays important role in the maintenance of spindle structure and function; however, the underlying mechanism remains largely undefined. Here we demonstrated that MERIT40, a core component of BRISC complex, directly interacts with the RXXPEG motif in the ARC-V domain of Tankyrase1(TNKS1). Mutation of the RXXPEG motif in the MERIT40 (R28A) disrupted its interaction with TNKS1. Consistent with these data, R28A mutant cells displayed multiple mitotic defects including aberrant spindle assembly and chromosome misalignment. These results support a critical role of RXXPEG motif of MERIT40 in BRISC-mediated regulation of TNKS1 function during spindle assembly.

Gastric cancer (GC) is one of the leading causes of cancer-related deaths throughout China and worldwide. The discovery of microRNAs (miRNAs) has provided a new opportunity for developing diagnostic biomarkers and effective therapeutic targets in GC. By performing microarray analyses of benign and malignant gastric epithelial cell lines (HFE145, NCI-N87, MKN28, RF1, KATO III and RF48), 16 significantly dysregulated miRNAs were found. 11 of these were validated by real-time qRT-PCR. Based on miRWalk online database scans, 703 potential mRNA targets of the 16 miRNAs were identified. Bioinformatic analyses suggested that these dysregulated miRNAs and their predicted targets were principally involved in tumor pathogenesis, MAPK signaling, and apoptosis. Finally, miRNA-gene network analyses identified miRNA-125b as a crucial miRNA in GC development. Taken together, these results develop a comprehensive expression and functional profile of differentially expressed miRNAs related to gastric oncogenesis. This profile may serve as a potential tool for biomarker and therapeutic target identification in GC patients.

The microtubule-associated protein BRISC regulates the interaction of NuMA with dynein and importin-β by removing K63-linked polyubiquitin chains from NuMA, thereby promoting proper bipolar spindle assembly., Deubiquitinating enzymes (DUBs) negatively regulate protein ubiquitination and play an important role in diverse physiological processes, including mitotic division. The BRCC36 isopeptidase complex (BRISC) is a DUB that is specific for lysine 63–linked ubiquitin hydrolysis; however, its biological function remains largely undefined. Here, we identify a critical role for BRISC in the control of mitotic spindle assembly in cultured mammalian cells. BRISC is a microtubule (MT)-associated protein complex that predominantly localizes to the minus ends of K-fibers and spindle poles and directly binds to MTs; importantly, BRISC promotes the assembly of functional bipolar spindle by deubiquitinating the essential spindle assembly factor nuclear mitotic apparatus (NuMA). The deubiquitination of NuMA regulates its interaction with dynein and importin-β, which are required for its function in spindle assembly. Collectively, these results uncover BRISC as an important regulator of the mitotic spindle assembly and cell division, and have important implications for the development of anticancer drugs targeting BRISC.

Invasion and metastasis are critical pathological and mortal processes in esophageal squamous cell carcinoma (ESCC). Novel drugs, targeting the two cancer migration stages, will augment the treatment options for ESCC therapy and improve overall survival. A novel natural macrolide F806 specifically promotes apoptosis of various ESCC cells. However, whether F806 can inhibit metastasis of ESCC cells needs further evaluation. Here, our data showed that F806 inhibits dynamic F-actin assembly and then suppresses the migration of ESCC cells in vitro and their invasion and metastasis in vivo. The correlation between cancer migration and actin cytoskeleton assembly was consistent with the ability of F806 to prevent the aggregation of Paxillin, an essential protein for focal adhesion formation through binding to the ends of actin filaments. Furthermore, F806 downregulated the expression and activity of the Rho family proteins cell division cycle 42 (CDC42), RAC family small GTPase 1 (RAC1), and RAS homolog family member A (RHOA). Taken together, these results suggest that F806 can suppress cancer invasion and metastasis via interrupting the assembly of migration components involving F-actin.

The current study is to develop a gentle and efficient method for purification of fibroblast-activation protein positive (FAP+) cancer-associated fibroblasts (CAFs) from tumor tissues. Fresh tissues were isolated from BALB/c-Nude mice bearing human liver cancer cell line (HepG2), fully minced and separated into three parts, and digested with trypsin digestion and then treated with collagenase type IV once, twice, or thrice, respectively. Finally, the cells were purified by using FAP magnetic beads. The isolated CAFs were grown in culture medium and detected for the surface expression of fibroblast-activation protein (FAP). The number of adherent cells which were obtained by digestion process with twice collagenase type IV digestion was (5.99±0.18) × 104, much more than that with the only once collagenase type IV digestion (2.58±0.41) × 104 (P<0.0001) and similar to thrice collagenase type IV digestion. The percentage of FAP+ CAFs with twice collagenase type IV digestion (38.5%) was higher than that with the only once collagenase type IV digestion (20.0%) and little higher than thrice collagenase type IV digestion (37.5%). The FAP expression of CAFs was quite different from normal fibroblasts (NFs). The fibroblasts isolated by the innovation are with high purity and being in wonderful condition and display the features of CAFs.

Circular RNAs (circRNAs) are a large class of endogenous RNAs, formed by exon skipping or back-splicing events as covalently closed loops, which are expressed abundantly in mammalian cells. Although their biological functions remain largely unknown, recent studies show that circRNAs have three main functions in mammalian cells. First, circRNAs can regulate transcription and RNA splicing. Second, circRNAs function as microRNA (miRNA) sponges. Third, they can be translated into protein driven by N6-methyladenosine modification. Taking advantage of RNA sequencing (RNA-seq) technology, the expressions of circRNAs were found to be dysregulated in all types of cancer cell lines, tumor tissues, and even plasma samples from patients, which correlated with certain clinical characteristics, suggesting the potential roles of circRNAs in tumor progression. Considering their conserved sequences and stable structures, circRNAs were deemed to be promising biomarkers for the early diagnosis and prognosis prediction of cancer. In this review, we describe briefly the formation and properties of circRNAs, and focus mainly on recent progress in research into their function, regulation, and clinical relevance in different cancers.

DNA interstrand crosslinks (ICLs) are extremely deleterious lesions that are repaired by homologous recombination (HR) through coordination of Fanconi anemia (FA) proteins and breast cancer susceptibility gene 1 (BRCA1) product, but the exact role these proteins have remains unclear. Here we report that FANCG was modified by the addition of lysine63-linked polyubiquitin chains (K63Ub) in response to DNA damage. We show that FANCG K63Ub was dispensable for monoubiquitination of FANCD2, but was required for FANCG to interact with the Rap80-BRCA1 (receptor-associated protein 80-BRCA1) complex for subsequent modulation of HR repair of ICLs induced by mitomycin C. Mutation of three lysine residues within FANCG to arginine (K182, K258 and K347, 3KR) reduced FANCG K63Ub modification, as well as its interaction with the Rap80-BRCA1 complex, and therefore impeded HR repair. In addition, we demonstrated that K63Ub-modified FANCG was deubiquitinated by BRCC36 complex in vitro and in vivo. Inhibition of BRCC36 resulted in increased K63Ub modification of FANCG. Taken together, our results identify a new role of FANCG in HR repair of ICL through K63Ub-mediated interaction with the Rap80-BRCA1 complex.

The aim of the current study was to investigate the expression of cyclin-dependent kinase 3 (CDK3) in human nasopharyngeal carcinoma (NPC) and to evaluate its association with the clinicopathological characteristics of patients with NPC. CDK3 expression was examined in three NPC cell lines and one nasopharyngeal epithelial cell line by western blot analysis and in 94 specimens of NPC and 40 specimens of inflamed nasopharyngeal tissue by immunohistochemistry staining. CDK3 was overexpressed in the three NPC cell lines, 5-8F, CNE1 and CNE2, compared with the NP-69 nasopharyngeal epithelial cell line, and was primarily expressed in the cytoplasm. The frequency of CDK3 expression was significantly higher in NPC specimens (67%) compared with inflamed nasopharyngeal tissue specimens (12.5%; P

In order to increase freeze-salt scaling resistance of concrete, effect of water-binder ratio, fly ash, slag and silica fume on freeze-salt scaling resistance are researched according to the CDF method. The results show that the scaled mass and the dynamic modulus of elasticity loss rate of the concrete are reduced with decreasement of water-binder ratio. When mineral admixture compound is added into concrete the scaled mass and the dynamic modulus of elasticity loss rate are also reduced. Compared with fly ash and slag the trend is more obvious as result of fume and slag added. The scaled mass and the loss rate of dynamic elasticity modulus are slightly reduced with decreasement of fly ash and slag. But the scaled mass and the loss rate of dynamic elasticity modulus are obviously reduced with increasement of silica fume and slag.

The addition of bevacizumab (BEV) to cytotoxic chemotherapy regimens (CTX) was believed to be effective; however, its magnitude of benefits is still controversial. So a meta-analysis and systematic review seems to be necessary.PubMed and the Cochrane library were systematically searched. All relevant citations comparing CTX with/without BEV were considered for inclusion. Sensitivity and meta-regression analysis were performed to identify potential confounders. All pooled estimates were performed using a random-effects model. All statistical analyses were performed by StataSE 12.0.The search strategy identified 10 eligible random control trials (RCTs) (n=1366). In our pooled estimates, the additional benefits of BEV to CTX were identified in overall survival (OS) hazard ratio (HR, 0.76; 95% CI, 0.69 to 0.82) and progression-free survival (PFS) (HR, 0.56; 95% CI, 0.51 to 0.60), and prolonged survival duration were also identified for OS (18.2 vs. 16.3, p=0.0003) and PFS (8.9 vs. 6.5, p0.001). Subgroup analyses stratified by CTX was also performed, evident benefits of additional BEV in OS and PFS can be identified in all subgroups, except for the CTX containing capecitabine in OS. Moreover, the increased rate of incidence was also identified in hypertension, thrombosis, proteinuria, gastrointestinal perforation, and fatigue.BEV, acting as a targeted agent to CTX, its additional benefit to CTX is at the cost of increased toxicity.

Background: Tumor vessels can potentially serve as diagnostic, prognostic and therapeutic targets for solid tumors. Fluorescent dyes are commonly used as biological indicators, while photobleaching seriously hinders their application. In this study, we aim to generate a fluorescent silica nanoparticles (FSiNPs) theranostic system marked by the mouse endgolin (mEND) aptamer, YQ26. Methods: A highly specific YQ26 was selected by using gene-modified cell line-based SELEX technique. FSiNPs were prepared via the reverse microemulsion method. The YQ26-FSiNPs theranostic system was developed by combining YQ26 with the FSiNPs for in vivo tumor imaging, treatment and monitoring. Results: Both in vitro experiments (i.e. cellular and tumor tissue targeting assays) and in vivo animal studies (i.e. in vivo imaging and antitumor efficacy of YQ26-FSiNPs) clearly demonstrated that YQ26-FSiNPs could achieve prominently high targeting efficiency and therapeutic effects via aptamer YQ26-mediated binding to endoglin (END) molecule. Conclusion: This simple, sensitive, and specific YQ26-FSiNPs theranostic system has a great potential for clinical tumor targeting imaging and treatment.

The active screen ion sulphurized layer was prepared on the surface of CrMoCu alloy cast iron by using active screen ion sulphurizing technology. Its key composition is FeS. Under epinoc grease lubrication condition, the friction factor and the wear volume of the active screen ion sulphurized layer is 24% and 40% lower than that of the plain surface and close to that of the sulphurized layer, respectively. The active screen ion sulphurized layer has excellent self-lubrication property, and its loose and porous structure tending to store the grease, which make it possess excellent friction reduction and wear resistance performances.

Fusion proteins containing the amino terminus of mixed lineage leukemia (MLL) are common in acute lymphoblastic leukemia (ALL) due to translocations. The MLL-AF4 fusion protein is generated by the translocation t(4;11)(q21;q23), and t(4;11)-positive ALL patients (MLL-AF4 ALL), have a notoriously poorer prognosis compared with patients with other MLL-associated leukemias. The detailed role of this fusion protein in leukemogenesis is not well understood. MicroRNAs (miRNAs) targeting the AF4 3' untranslated regions may modulate MLL-AF4 fusion protein levels, raising the question of whether regulation of these miRNAs are involved in the progression of MLL-AF4 ALL. In this study, we show that miR-143 was identified as a regulator of MLL-AF4 expression in MLL-AF4 ALL samples. Restoration of miR-143 in MLL-AF4-positive RS4;11 and MV4-11 cells induced apoptosis, negatively contributing to leukemia cell growth by reducing MLL-AF4 fusion protein levels. Furthermore, miR-143 was epigenetically repressed by promoter hypermethylation in MLL-AF4-positive primary blasts and cell lines, but not in normal bone marrow cells and MLL-AF4-negative primary blasts, which was directly associated with expression of the MLL-AF4 oncogene. This is the first study to show that miR-143 functions as a tumor suppressor in MLL-AF4 B-cell ALL. These data reveal the therapeutic promise of upregulating miR-143 expression for MLL-AF4 B-cell ALL.

Histone H2AX is a histone H2A variant that is ubiquitously expressed throughout the genome. It plays a key role in the cellular response to DNA damage and has been designated as the histone guardian of the genome. Histone H2AX deficiency decreases genomic stability and increases tumor susceptibility of normal cells and tissues. However, the role of histone H2AX phosphorylation in malignant transformation and cancer development is not totally clear. Herein, we found that ribosomal S6 kinase 2 (RSK2) directly phosphorylates histone H2AX at Ser139 and also at a newly discovered site, Ser16. Epidermal growth factor (EGF)–induced phosphorylation of histone H2AX at both sites was decreased in RSK2 knockout cells. Phosphorylated RSK2 and histone H2AX colocalized in the nucleus following EGF treatment, and the phosphorylation of histone H2AX by RSK2 enhanced the stability of histone H2AX and prevented cell transformation induced by EGF. RSK2 and DNA-PK, but not ATM or ATR, are required for EGF-induced phosphorylation of H2AX at Ser139; however, only RSK2 is required for phosphorylation of H2AX at Ser16. Phosphorylation of histone H3 was suppressed in cells expressing wild-type H2AX compared with H2AX knockout (H2AX−/−) cells. EGF-associated AP-1 transactivation activity was dramatically lower in H2AX−/− cells overexpressing wild-type H2AX than H2AX−/− cells expressing mutant H2AX-AA. Thus, the RSK2/H2AX signaling pathway negatively regulates the RSK2/histone H3 pathway and therefore maintains normal cell proliferation. Cancer Res; 71(2); 393–403. ©2011 AACR.

c-Jun is a component of the activator protein-1 (AP-1) complex, which plays a crucial role in the regulation of gene expression, cell proliferation, and cell transformation, as well as cancer development. Herein, we found that cyclin-dependent kinase (Cdk)-3, but not Cdk2 or c-Jun NH2-terminal kinase, is a novel kinase of c-Jun induced by stimulation with growth factors such as epidermal growth factor (EGF). Cdk3 was shown to phosphorylate c-Jun at Ser63 and Ser73 in vitro and ex vivo. EGF-induced Cdk3 activation caused c-Jun phosphorylation at Ser63 and Ser73, resulting in increased AP-1 transactivation. Ectopic expression of Cdk3 resulted in anchorage-independent cell transformation of JB6 Cl41 cells induced by EGF and foci formation stimulated by constitutively active Ras (RasG12V), which was mediated by AP-1 in NIH3T3 cells. These results showed that the Cdk3/c-Jun signaling axis plays an important role in EGF-stimulated cell proliferation and cell transformation. [Cancer Res 2009;69(1):272–81]

Cyclin-dependent kinase (cdk)-3, a member of the cdk family of kinases, plays a critical role in cell cycle regulation and is involved in G0-G1 and G1-S cell cycle transitions. However, the role of cdk3 in cell proliferation, as well as cell transformation, is not yet clearly understood. Here, we report that the protein expression level of cdk3 is higher in human cancer cell lines and human glioblastoma tissue compared with normal brain tissue. Furthermore, we found that cdk3 phosphorylates activating transcription factor 1 (ATF1) at serine 63 and enhances the transactivation and transcriptional activities of ATF1. Results also indicated that siRNA directed against cdk3 (si-cdk3) suppresses ATF1 activity, resulting in inhibition of proliferation and growth of human glioblastoma T98G cells in soft agar. Importantly, we showed that cdk3 enhances epidermal growth factor–induced transformation of JB6 Cl41 cells and si-cdk3 suppresses RasG12V/cdk3/ATF1–induced foci formation in NIH3T3 cells. These results clearly showed that the cdk3-ATF1 signaling axis is critical for cell proliferation and transformation. [Cancer Res 2008;68(18):7650–60]

Solar UV irradiation is an important carcinogen that leads to the development of skin cancer, which is the most common human cancer. However, the receptors that mediate UV-induced skin carcinogenesis have not yet been unequivocally identified. Here we showed that UV irradiation directly activates cannabinoid receptors 1 and 2 (CB1/2). Notably, our data indicated that the absence of the CB1/2 receptors in mice results in a dramatic resistance to UVB-induced inflammation and a marked decrease in UVB-induced skin carcinogenesis. A marked attenuation of UVB-induced activation of mitogen-activated protein kinases and nuclear factor- κB was associated with CB1/2 deficiency. These data provide direct evidence indicating that the CB1/2 receptors play a key role in UV-induced inflammation and skin cancer development. [Cancer Res 2008;68(10):3992–7]

Insulin-like growth factor-I receptor (IGF-IR) has been implicated in cancer pathophysiology. Furthermore, impairment of IGF-IR signaling in various cancer cell lines caused inhibition of the transformed phenotype as determined by the inhibition of colony formation in soft agar and the inhibition of tumor formation in athymic nude mice. Thus, the IGF-IR might be an attractive target for cancer prevention. We showed that the tea polyphenol, (−)−epigallocatechin-3-gallate (EGCG), is a small-molecule inhibitor of IGF-IR activity (IC50 of 14 μmol/L). EGCG abrogated anchorage-independent growth induced by IGF-IR overexpression and also prevented human breast and cervical cancer cell phenotype expression through inhibition of IGF-IR downstream signaling. Our findings are the first to show that the IGF-IR is a novel binding protein of EGCG and thus may help explain the chemopreventive effect of EGCG on cancer development. (Cancer Epidemiol Biomarkers Prev 2007;16(3):598–605)

Senescence, a terminal cell proliferation arrest that is caused by a variety of cellular stresses such as telomere erosion, DNA damage and oncogenic signaling, is classically considered a tumor defense barrier. However, the mechanism by which cancer cells overcome senescence is undetermined. In this study, the gene expression array data of esophageal squamous cell carcinoma (ESCC) was compared with paired normal tissues and showed that a cohort of genes, including proteinases, chemokines and inflammation factors, are upregulated in ESCC, which exhibits the senescence-associated secretory phenotype. In addition, reverse transcription-quantitative polymerase chain reaction was used to demonstrate that gender determining region Y-box 4 (SOX4) is upregulated in ESCC, and that its expression is inversely correlated with senescence markers. In addition, the knockdown of SOX4 expression by short hairpin RNA decreases ESCC cell proliferation and enhances doxorubicin-induced cell senescence. These results reveal the presence of a senescent microenvironment in ESCC, and suggest an important antisenescence role of SOX4 in ESCC progression.

Development of reliable vectors is a major challenge in gene therapy. Previous gene transfer methods using non-viral vectors, such as liposomes or nanoparticles, have resulted in relatively low levels (35 to approximately 50%) of gene expression. We have developed a silicon nanoparticle (SNAP) system, a novel non-viral vector, for DNA transfer into cells. SNAP was synthesized chemically and modified with sodium chloride or sodium iodide. Electronmicroscopy of SNAP and fluorescence microscopy of fluorescence-labeled SNAP revealed that they were generated uniformly, had diameters of 10-100 nm, and showed a better efficiency (about 70%) of DNA transfection into cells as well as protection of DNA against degradation. The microscopy also demonstrated the adhesion of SNAP with HT1080 cell surface and entry of SNAP into the cells without cytotoxicity. Intravenous and/or intra-abdominal administration of the SNAP to mice revealed the accumulation of SNAP in the cells of the brain, liver, spleen, lung, kidney, intestine, prostate and the testis without any pathological cell changes or mortality, suggesting that they passed through the blood-brain, blood-prostate, and blood-testis barriers. These findings indicate that the SNAP generated has good biological characteristics as a potential promising vector for gene transfer, gene therapy and drug delivery.

Low fusion efficiency and nominal activity of fusion cells (FCs) restrict the clinical application of dendritic cell (DC)/tumor fusion cells. Collagen I (Col I) is an interstitial collagen with a closely-knit structure used to repair damaged cell membranes. This study evaluated whether Col I could improve the fusion efficiency of polyethylene glycol (PEG)-induction and enhance the immunogenicity of fusion vaccine. DC/B16 melanoma and controlled DC/H22 hepatoma cell fusions were induced by PEG with or without Col I. Col I/PEG treatment increased the levels of DC surface molecules and the secretion of lactate, pro- and anti-inflammatory cytokines in fusion cells. Col I/PEG-treated FCs enhanced T-cell proliferation and cytotoxic T lymphocyte activity. The Col I-prepared fusion vaccine obviously suppressed tumor growth and prolonged mice survival time. Thus Col I treatment could significantly improve the efficiency of PEG-induced DC/tumor fusion and enhance the anticancer activity of the fusion vaccine. This novel fusion strategy might promote the clinical application of DC/tumor fusion immunotherapy.

Aberrant PDGF-PDGFR signaling and its effects on downstream effectors have been implicated in glioma development. A crucial AKT regulator, ACK1 (TNK2) has been shown to be a downstream mediator of PDGF signaling; however, the exact underlying mechanisms in gliomas remain elusive. Here, we report that in glioma cells, PDGFR-β activation enhanced the interaction between ACK1 and AKT, resulting in AKT activation. PDGF treatment consistently promoted the formation of complexes containing PDGFR-β and ACK1. Mutational analysis suggested that Y635 of ACK1 is a PDGFR-β phosphorylation site and that the ACK1 Y635F mutant abrogated the sequential activation of AKT. Moreover, PDK1 interacted with ACK1 during PDGF stimulation, which is required for the binding of ACK1 to PDGFR-β. Further mutational analysis showed that T325 of ACK1 was crucial for the ACK1 and PDK1 interaction. ACK1 Y635F or T325A mutants abolished PDGFR-β-induced AKT activation, the subsequent nuclear translocation of β-catenin and the expression of cyclin D1. Glioma cell cycle progression, proliferation and tumorigenesis were accordingly blocked by ACK1 Y635F or T325A. In glioblastoma multiforme samples from 51 patients, increased ACK1 tyrosine phosphorylation correlated with upregulated PDGFR-β activity and AKT activation. Taken together, our data demonstrate that ACK1 plays a pivotal role in PDGF-PDGFR-induced AKT signaling in glioma tumorigenesis. This knowledge contributes to our understanding of glioma progression and may facilitate the identification of novel therapeutic targets for future glioma treatment.

To investigate the expression profile and the clinical significance of CDK3 in the tissues of nasopharyngeal carcinoma.Immunohistochemistry was utilized to detect the expression of CDK3 in 94 cases of NPC and 40 cases of nasopharyngeal inflammation.CDK3 was highly expressed in NPC cell lines. Immunohistochemistry showed that CDK3 was mostly expressed in the cytoplasm. The positive expression rate of NPC was 67% and that of nasopharyngeal inflammation was only 12. 5%. The difference between these two groups was highly statistically significant. The CDK3 expression in NPC was related to the TNM clinical staging of NPC (P0.01).The expression levels of CDK3 were obviously higher in NPC tissues. The CDK3 expression in NPC was related to the TNM clinical staging of NPC. It suggests that CDK3 expression may play an important role in the occurrence and development of nasopharyngeal carcinoma.

The corrosion currents of steel bar in concrete with three W/B and four chloride contents after surface-painting two migrating corrosion inhibitors (PCI-2015 and MCI-A) 14d to 150d in atmospheric condition were measured. The results showed that the corrosion current density (I corr) of steel bar reduced to 0.1 μA.cm-2 from the initial highest 3.833 μA.cm-2 (W/B=0.65, NaCl-1%) after surface-painting PCI-2015 14 d, and the I corr was still lower than 0.1 μA.cm-2 until 150d. The compressive strength and chloride migration coefficient of concrete specimens were tested. The possible reasons of the mechanisms of durability improvement for reinforced concrete by applying PCI-2015 inhibitor were PCI-2015 may be reacted with calcium hydroxide in cement concrete and lots of inhibitor particles may be adsorbed on the active sites first and then a stable protective layer may be formed. The I corr of steel bars in a hydraulic aqueduct concrete structure after painting PCI-2015, MCI-A (the United States) and MCI-B (Europe) during 6 months was monitored by Gecor 8 tester. The results showed that the average values of I corr of steel bars after painting the PCI-2015 150d fulfilled the specification requirements in "Design code for concrete structure strengthening (E.3) "(GB 50367-2013).

Various types of post-translational modifications of the histone tails have been revealed, but a few modifications have been found within the histone core sequences. Histone core post-translational modifications have the potential to modulate nucleosome structure and DNA accessibility. Here, we studied the histone H2B core domain and found that phosphorylation of H2B serine 32 occurs in normal cycling and mitogen-stimulated cells. Notably, this phosphorylation is elevated in skin cancer cell lines and tissues compared with normal counterparts. The JB6 Cl41 mouse skin epidermal cell line is a well established model for tumor promoter-induced cell transformation and was used to study the function of H2B during EGF-induced carcinogenesis. Remarkably, cells overexpressing a nonphosphorylatable H2BS32A mutant exhibited suppressed growth and EGF-induced cell transformation, possibly because of decreased activation of activator protein-1, compared with control cells overexpressing wild type H2B. We identified ribosomal S6 kinase 2 (RSK2) as the kinase responsible for H2BS32 phosphorylation. Serum-starved JB6 cells contain very little endogenous H2BS32 phosphorylation, and EGF treatment induced this phosphorylation. The phosphorylation was attenuated in RSK2 knock-out MEFs and RSK2 knockdown JB6 cells. Taken together, our results demonstrate a novel role for H2B phosphorylation in cell transformation and show that H2BS32 phosphorylation is critical for controlling activator protein-1 activity, which is a major driver in cell transformation.

MST1 (mammalian STE20-like kinase 1) is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. Overexpression of MST1 induces apoptotic morphological changes such as chromatin condensation, but the mechanism is not clear. Here we show that MST1 induces apoptotic chromatin condensation through its phosphorylation of histone H2AX at Ser-139. During etoposide-induced apoptosis in Jurkat cells, the cleavage of MST1 directly corresponded with strong H2AX phosphorylation. In vitro kinase assay results showed that MST1 strongly phosphory-lates histone H2AX. Western blot and kinase assay results with a mutant S139A H2AX confirmed that MST1 phos-phorylates H2AX at Ser-139. Direct binding of MST1 and H2AX can be detected when co-expressed in HEK293 cells and was also confirmed by an endogenous immunoprecipitation study. When overexpressed in HeLa cells, both the MST1 full-length protein and the MST1 kinase domain (MST1-NT), but not the kinase-negative mutant (MST1-NT-KN), could induce obvious endogenous histone H2AX phosphorylation. The caspase-3 inhibitor benzyloxycarbonyl-DEVD-fluoromethyl ketone (Z-DEVD-fmk) attenuates phosphorylation of H2AX by MST1 but cannot inhibit MST1-NT-induced histone H2AX phosphorylation, indicating that cleaved MST1 is responsible for H2AX phosphory-lation during apoptosis. Histone H2AX phosphorylation and DNA fragmentation were suppressed in MST1 knockdown Jurkat cells after etoposide treatment. Taken together, our data indicated that H2AX is a substrate of MST1, which functions to induce apoptotic chromatin condensation and DNA fragmentation.

Extracellular signal-regulated kinase 8 (ERK8), a recently discovered member of the mitogen-activated protein kinase protein family, has been less studied than other family members, leaving its physiologic functions mostly unknown. The biological consequences of overexpression of ERK8 in JB6 Cl41 epidermal skin cells or knockdown of ERK8 in HCT15 colorectal cancer cells was studied. Kinase assays and transient transfection experiments were performed to study the signaling pathway between ERK8 and c-Jun. We found that ERK8 is relatively highly expressed in HCT15 human colorectal cancer cells and plays an important role in the promotion and progression of colorectal cancer. ERK8 promoted neoplastic transformation, and knockdown of ERK8 in HCT15 colorectal cancer cells reduced the tumorigenic properties of these cell lines. Furthermore, a direct interaction between ERK8 and c-Jun was shown. With epidermal growth factor treatment, overexpression of ERK8 in JB6 Cl41 cells caused an increased phosphorylation of c-Jun at Ser63 and Ser73, resulting in increased activator protein-1 transactivation. In contrast, knockdown of ERK8 in HCT15 colorectal cancer cells blocked c-Jun phosphorylation. The interaction between ERK8 and c-Jun seems to increase the tumorigenic properties of HCT15 colorectal cancer cells. Thus, ERK8-regulated signaling might serve as a potential therapeutic target in colorectal cancer. Cancer Res; 70(8); 3218–27. ©2010 AACR.

The p90 ribosomal protein kinase 2 (RSK2) is a highly expressed Ser/Thr kinase activated by growth factors and is involved in cancer cell proliferation and tumor promoter-induced cell transformation. RSK2 possesses two non-identical kinase domains, and the structure of its N-terminal domain (NTD), which is responsible for phosphorylation of a variety of substrates, is unknown. The crystal structure of the NTD RSK2 was determined at 1.8 A resolution in complex with AMP-PNP. The N-terminal kinase domain adopted a unique active conformation showing a significant structural diversity of the kinase domain compared to other kinases. The NTD RSK2 possesses a three-stranded betaB-sheet inserted in the N-terminal lobe, resulting in displacement of the alphaC-helix and disruption of the Lys-Glu interaction, classifying the kinase conformation as inactive. The purified protein was phosphorylated at Ser227 in the T-activation loop and exhibited in vitro kinase activity. A key characteristic is the appearance of a new contact between Lys216 (betaB-sheet) and the beta-phosphate of AMP-PNP. Mutation of this lysine to alanine impaired both NTDs in vitro and full length RSK2 ex vivo activity, emphasizing the importance of this interaction. Even though the N-terminal lobe undergoes structural re-arrangement, it possesses an intact hydrophobic groove formed between the alphaC-helix, the beta4-strand, and the betaB-sheet junction, which is occupied by the N-terminal tail. The presence of a unique betaB-sheet insert in the N-lobe suggests a different type of activation mechanism for RSK2.

Blockade of the transient receptor potential channel vanilloid subfamily 1 (TRPV1) is suggested as a therapeutic approach to pain relief. However, TRPV1 is a widely expressed protein whose function might be critical in various nonneuronal physiologic conditions. The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that is overexpressed in many human epithelial cancers and is a potential target for anticancer drugs. Here, we show that TRPV1 interacts with EGFR, leading to EGFR degradation. Notably, the absence of TRPV1 in mice results in a striking increase in skin carcinogenesis. The TRPV1 is the first membrane receptor shown to have a tumor-suppressing effect associated with the down-regulation of another membrane receptor. The data suggest that, although a great deal of interest has focused on TRPV1 as a target for pain relief, the chronic blockade of this pain receptor might increase the risk for cancer development. [Cancer Res 2009;69(3):905–13]

The ribosomal S6 kinase 2 (RSK2), a member of the p90RSK (RSK) family of proteins, is a widely expressed serine/threonine kinase that is activated by extracellular signal-regulated kinase 1/2 and phosphoinositide-dependent kinase 1 in response to many growth factors and peptide hormones. Its activation signaling enhances cell survival. However, the roles of RSK2 in cell transformation have not yet been elucidated. Here, we found that RSK2 is a critical serine/threonine kinase for the regulation of cell transformation. When cells were stimulated with tumor promoters, such as epidermal growth factor (EGF) or 12-O-tetradecanoylphorbol-13-acetate (TPA), phosphorylation of RSK was increased within 5 min. Cell proliferation was suppressed in RSK2−/− mouse embryonic fibroblasts (MEFs) compared with RSK2+/+ MEFs. Moreover, RSK2−/− MEFs accumulated at the G1 phase of the cell cycle under normal cell culture conditions as well as after stimulation with EGF or TPA. In the anchorage-independent cell transformation assay (soft agar), stable expression of RSK2 in JB6 cells significantly enhanced colony formation in either the presence or absence of tumor promoters. Furthermore, knockdown of RSK2 with small interfering RNA-RSK2 suppressed constitutively active Ras (RasG12V)-induced foci formation in NIH3T3 cells. In addition, kaempferol, an inhibitor of RSK2, suppressed EGF-induced colony formation of JB6 Cl41 cells in soft agar, which was associated with inhibition of histone H3 phosphorylation (Ser10). These results showed that RSK2 is a key regulator for cell transformation induced by tumor promoters such as EGF and TPA. [Cancer Res 2007;67(17):8104–12]

Hereditary nonpolyposis colorectal cancer (HNPCC) is one of the most common hereditary colon cancer syndromes accounting for 1%-5% of all colorectal cancer cases. Germline mutations in at least five genes coding for DNA mismatch repair (MMR) proteins are associated with the clinical phenotype of HNPCC. More than 400 MMR mutations have been identified in HNPCC patients, and about 40% of mutations affect MSH2 gene including nucleotide substitutions, deletions, and insertions. Only a few mutations have been reported in Chinese families.A Chinese family with HNPCC was collected and peripheral blood of individuals from the family was obtained. Mutation analysis was performed on genomic DNA.The family fulfilled Amsterdam criteria I, and 17 people out of 31 were diagnosed as malignant tumor for 21 times. Twelve people (70.6%) had rectal cancer, and the onset age was young with an average of 42.9 years old. Right side colon cancer was common in the family. A novel duplication mutation of four nucleotides in exon 7 MSH2 (MSH2: c.1215_1218dupCCGA) was found, which result in a premature stop 10 codons downstream in MSH2 (p.L407fsX417) in the family. Site-specific PCR was applied to the pre-symptomatic diagnosis.This novel genomic mutation MSH2 was confirmed to be pathogenic, and polymerase chain reaction with modified primer was successfully applied to the pre-symptomatic diagnosis. These data expand the spectrum MSH2 mutations causing HNPCC.

To establish a genetic diagnosis method for a novel MSH2 mutation.A specific primer on the mutated site of MSH2 was synthesized and PCR was conducted using the specific primer and another downstream primer. PCR products were electrophoresed and then the carriers with the novel gene mutation of the carriers or non-carriers were identified.MSH2 in a hereditary nonpolyposis colorectal cancer family were successfully found.The method is effective and simple for genetic diagnosis of the novel mutation in MSH2.

To evaluate the disease-causing gene and phenotypic characters of a large family with autosomal dominant retinitis pigmentosa (adRP).Disease status and associated ocular abnormalities of eight patients and six unaffected members who represent different generations of this family were assessed by measurement of visual psychophysics, full-field and multifocal electrophysiology (ERG and mfERG) and funds fluorescent angiography (FFA). The DNA samples of nineteen patients and fifteen unaffected individuals in this family were examined by Genome scanning, linkage analysis and mutation detection to identify coding sequence changes.A case with variable, early onset night blindness before 10 years and visual field loss in their teens was found. Macular dystrophy, progressing to a retinitis pigmentosa phenotype was demonstrated in most adult cases. Both a-wave and b-wave amplitudes of photopic and scotopic full-field ERG were marked reduced and nearly non-detectable, demonstrating severe damage of photoreceptor systems. There were two obligate gene carriers in the family which remained asymptomatic in the clinical. But one of them was found with a minimal RP characteristic and the other was normal by examination of fundus and ERG. An unreported splicing site mutation (IVS5-1GA) was identified in intron 5- acceptor site of PRPF-31 gene on chromosome 19. ERG and molecular genetic findings were consistent with the reclassification of this disease as an autosomal dominant RP.It is a novel splicing site mutation that IVS5-1GA of D19S418 site in PRFP31, the relative phenotypes by which main displayed type I/diffuse has variable expressivity and complex phenotype.

In order to obtain a simple,fast,accurate and low-cost diagnosis method of fragile X syndrome, cytogenetic tests and molecular genetic tests were carried out with direct amplification of (CGG)(n) repeat sequence in 5' terminal of FMR1 gene by PCR and the cDNA sequence of FMR1 by RT-PCR from six mental retardation pedigrees. The proband of pedigree A with high expression of fragile X chromosome(35/273) was detected to be a full mutation patient of fragile X syndrome by the molecular genetic test. There is no expression of fragile X chromosome in the proband and his mother of pedigree B, which was further confirmed as a non-fragile X pedigree by the molecular genetic test. A male foetus of the pedigree C has fragile X chromosome(5/93), but the proband and his mother has no fragile X chromosome. By further detection using molecular genetic test, the male foetus is a full mutation patient of fragile X syndrome, his mother is a permutated carrier, and his brother is a mosaic patient. The proband of pedigree D has high expression of fragile X chromosome (17%), his sister also has expression of fragile X chromosome (5%). By further detection with molecular genetic test, the proband is a full mutation patient of fragile X syndrome,and his sister is a mosaic patient. The probands of pedigrees E and F of the mother were found with suspicions fragile X chromosome, being confirmed as the non-fragile X pedigrees by the molecular genetic test. The conclusion is that the analysis test with direct amplification of 5'j(CGG)n repeat sequence and cDNA sequence in FMR1 gene is simple,fast,low-cost and can be applied in screening, diagnosis and prenatal diagnosis of fragile X syndrome.

The autosomal dominant form of retinitis pigmentosa (ADRP) can be caused by mutations in 13 genes and a further locus for which the gene remains to be identified. This study was intended to identify mutations in a large Chinese pedigree with ADRP.A genome scan was conducted in the family. The whole coding sequences and the intron-exon boundaries of candidate genes were amplified and sequenced. The reverse transcriptase polymerase chain reaction (RT-PCR) was performed to amplify the mutated mRNA.The strongest evidence of linkage was detected with three adjacent microsatellite markers genotyped between D19S902 and D19S210 on chromosome 19q13.33-13.43. Within the region, a single nucleotide change (GA) at position -1 of Intron 5 of PRPF31 was found. The consensus AG doublet of the Intron 5 splice acceptor was changed to AA. The mutation co-segregated with the disease phenotype, suggesting that it was the disease-causing mutation in this family. This splicing site mutation is predicted to cause an erroneous splicing of Exon 6. By RT-PCR, we found the mutated nucleotide of Intron 5 (A) and the first nucleotide of Exon 6 (G) was regarded as a new splice acceptor, resulting in 1 bp deletion of the first codon of Exon 6 (GAG-to-AG) at the mRNA level. This change led to a frameshift and truncated protein of 196 amino acids with 56 novel amino acids prior to a premature stop.A novel splicing mutation (IVS5-1GA) in the pre-mRNA splicing-factor gene PRPF31 causes retinitis pigmentosa in a large Chinese family. The mutation results in a truncated protein of PRPF31.