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Early diagnosis of hepatocellular carcinoma (HCC) lacks highly sensitive and specific protein biomarkers. Proteomics-driven discovery of tumor biomarkers is an important direction for omics study. Here, we described a staged mass spectrometry (MS)-based discovery-verification-validation proteomics workflow to explore serum proteomic biomarkers for HCC early diagnosis in 662 individuals (373 HCC patients and 289 non-HCC patients). Our workflow reproducibly quantified 451serum proteins using a data independent acquisition mass spectrometry (DIA-MS) strategy from discovery cohort, and proteins with significantly altered abundance in HCC were validated as candidates in an independent validation cohort using targeted proteomics based on parallel reaction monitoring (PRM). Machine learning models determined as P4 serum protein-panels (two serum proteomics biomarkers: HABP2, CD163 and two clinical used serum biomarkers: AFP, PIVKA-II) could clearly distinguish HCC patients from LC patients in an independent validation cohort (AUC 0.979, sensitivity 0.925, specificity 0.915), outperforming existing clinical prediction strategies (p

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Purpose:Intratumoral hepatitis B virus (HBV) integrations and mutations are related to hepatocellular carcinoma (HCC) progression. Circulating cell-free DNA (cfDNA) has shown itself as a powerful noninvasive biomarker for cancer. However, the HBV integration and mutation landscape on cfDNA remains unclear.Experimental Design:A cSMART (Circulating Single-Molecule Amplification and Resequencing Technology)-based method (SIM) was developed to simultaneously investigate HBV integration and mutation landscapes on cfDNA with HBV-specific primers covering the whole HBV genome. Patients with HCC (n = 481) and liver cirrhosis (LC; n = 517) were recruited in the study.Results:A total of 6,861 integration breakpoints including TERT and KMT2B were discovered in HCC cfDNA, more than in LC. The concentration of circulating tumor DNA (ctDNA) was positively correlated with the detection rate of these integration hotspots and total HBV integration events in cfDNA. To track the origin of HBV integrations in cfDNA, whole-genome sequencing (WGS) was performed on their paired tumor tissues. The paired comparison of WGS data from tumor tissues and SIM data from cfDNA confirmed most recurrent integration events in cfDNA originated from tumor tissue. The mutational landscape across the whole HBV genome was first generated for both HBV genotype C and B. A region from nt1100 to nt1500 containing multiple HCC risk mutation sites (OR > 1) was identified as a potential HCC-related mutational hot zone.Conclusions:Our study provides an in-depth delineation of HBV integration/mutation landscapes at cfDNA level and did a comparative analysis with their paired tissues. These findings shed light on the possibilities of noninvasive detection of virus insertion/mutation.

Heat from gas explosions affects the physical and mechanical properties of rock and reduces the stability of tunnels, such as in coal mines. This paper mainly studies the physical and mechanical properties of sandstone after heating and investigates the influences and mechanisms involved. Acoustic emission (AE) monitoring, X-ray diffraction (XRD), scanning electron microscopy (SEM) and polarized light microscopy are used to show that the uniaxial compressive strength (UCS) and peak strain of sandstone change significantly with temperature. In particular, the UCS decreases rapidly at 500 ~ 600 °C due to a transition from $${\alpha }$$ -quartz to $$\upbeta $$ -quartz. The XRD analysis shows that the physical and mechanical properties of sandstone are affected by its mineral composition, as physical and chemical reactions of quartz, muscovite and kaolinite at high temperatures weaken the sandstone. The AE signals increase with temperature and correspond well with observed loading stages, as they reflect the spatiotemporal evolution in internal micro-cracks. The degree of crack opening is low at low temperatures, then cracks become connected with increasing temperature, further explaining the changes in mechanical properties. Polarized light microscopy shows that grain size and pore characteristics are related to mechanical strength; generally, higher strength is associated with smaller grains with a less even size distribution. In addition, thermal and mechanical damage were quantitatively characterized by deriving thermal, comprehensive and dynamic damage factors. The results provide a theoretical reference for mining engineering in high-temperature environments.

To investigate the effects of Pien Tze Huang (PZH) on the migration and invasion of HCC cells and underlying molecular mechanism.Cell counting kit-8 (CCK-8) was applied to evaluate the cell viabilities of SMMC-7721, SK-Hep-1, C3A and HL-7702 (6 × 10PZH treatment significantly inhibited the migration, invasion, proliferation and promoted the apoptosis of HCC cells in vitro and in vivo (P0.01). Transcriptome analysis indicated that Hippo signaling pathway was associated with anti-metastasis function of PZH. Mechanical study showed PZH significantly inhibited the expressions of platelet derived growth factor receptor beta (PDGFRB), YAP, connective tissue growth factor (CCN2), N-cadherin, vimentin and matrix metallopeptidase 2 (MMP2, P0.01). Meanwhile, the phosphorylation of YAP was also enhanced by PZH treatment in vitro and in vivo. Furthermore, PZH played roles in inhibiting the transcriptional activity of YAP.PZH restrained migration, invasion and epithelial-mesenchymal transition of HCC cells through repressing PDGFRB/YAP/CCN2 axis.

The physical and mechanical properties of surrounding rock of high level radioactive waste repository are affected by the heat released by nuclide decay. In this paper, the effects of temperature on P-wave velocity, porosity and tensile strength are investigated, the microstructure of the specimens with different temperature is observed utilizing a polarizing microscope. Brazilian splitting tests on the heated specimens combined with the acoustic emission technology to monitor the rock deformation and failure. It is observed that the P-wave velocity of granite decreases with the increase of temperature. The porosity of granite changes with temperature and has obvious threshold that is the porosity of granite will increase rapidly when the temperature exceeds 500 °C. Besides, the tensile strength of granite shows a linear decrease with the increasing temperature. The results of acoustic emission tests and thin section observations indicate that temperature leads to an increase in the closure and unstable extension of cracks. The findings of the present research can provide reference for construction safety of the rock mass subjected to high temperature.

Privacy protection and message authentication issues in VANETs have received great attention in academia. Many authentication schemes in VANETs have been proposed, but most of them are based on classical difficult problems such as factorization in RSA setting or Elliptic Curve setting and are therefore not quantum resistant. If a quantum computer becomes available in the next few decades, the security of these schemes will be at stake. This paper presents a vehicular lattice-based direct anonymous attestation (V-LDAA) scheme adopting an optimized signature scheme based on automorphism stability which achieves postquantum security. A distributed pseudonym update and vehicle revocation mechanism based on the lattice is introduced in this paper, which means vehicles can update their pseudonyms and revoke the identity certificate by themselves without the need for pseudonym resolutions or CRLs checking. Compared with the existing lattice-based attestation schemes in VANETs, computation costs during signing and verification operations in V-LDAA are no longer related to the number of users, which makes it suitable for large-scale VANETs. Security analysis shows that V-LDAA resists TPM theft attacks and provides users with user-controlled anonymity, user-controlled unlinkability, and unforgeability against quantum adversaries. Experimental results show that V-LDAA reduces the blind signature size by 18%. The speed of blind signing is increased by 30%, and blind verification operation is accelerated 3 times compared with the existing lattice-based direct anonymous attestation (LDAA) scheme.

Myocardial ischemia-reperfusion injury (IRI) is a severe trauma which is characterized by inflammatory reaction, oxidative stress and cardiomyocyte apoptosis. Anesthetics such as sevoflurane have been proved to exhibit cardioprotective effect on IRI and the present study aimed to explore the underlying mechanism. In this study, H9C2 cells were randomly divided into the following groups: Control group; hypoxia/reoxygenation (H/R) group; 2.5% sevoflurane (Sev) 1 h group (H9C2 cells were exposed to 1 h of 2.5% sevoflurane 24 h before H/R); 2.5% Sev 2 h group (H9C2 cells were exposed to 2 h of 2.5% sevoflurane 24 h before H/R); 2.5% sevoflurane (Sev) 2 h + LY294002 group (H9C2 cells were pretreated with 10 μL LY294002 for 24 h before sevoflurane treatment). Cell proliferation and apoptosis were examined by CCK8 assay and Flow cytometry. Then, the expression levels of key proteins, including Bcl-2, Mcl-1, iNOS, p-AKT, t-AKT, PIM1, P-Bad, p-GSK3β, t-GSK3β and cyclinD1, were examined by western blot. Furthermore, nitric oxide (NO) concentration was detected with an ELISA kit, and TNF-α, IL-1β, IL-6 and IL-10 levels were examined by western blot. The CCK8 assay and flow cytometry results indicated that sevoflurane pretreatment reduced the apoptosis of H9C2 cells with H/R injury. In addition, sevoflurane pretreatment significantly inhibited the inflammatory injury induced by H/R. Furthermore, sevoflurane activated AKT/Pim-1 and AKT/GSK3β signaling pathway. These beneficial effects of sevoflurane were canceled by phosphoinositide-3-kinase inhibitor LY294002. In conclusion, these results verified that sevoflurane attenuates H/R-induced cardiomyocyte injury via AKT/Pim-1 and AKT/GSK3β signaling pathways.

The Wulonggou Pluton is located in Wulonggou area, eastern segment of the Eastern Kunlun Orogenic Belt, NW China, and consists of mainly alkali-feldspar granites covering an area of about 150 km2. Petrogenesis of these granitoids has been investigated through an integrated study of petrography, zircon U Pb ages, whole-rock geochemistry, and Hf Nd isotopic compositions. U Pb dating of magmatic zircons indicated these granites crystallized during 426–424 Ma in the middle Silurian. The granites display high SiO2 (75.26–77.55 wt%), K2O + Na2O (7.98–9.03 wt%), extremely low MgO (0.04–0.19 wt%), CaO (0.28–0.61 wt%), and TiO2 (0.05–0.09 wt%) contents showing metaluminous, calcic-alkali and ferroan features; enrichment in Rb and some HFSEs (Zr, U, Nb, Ta, and Y), depletion in Sr, Ba, P, and Ti, mostly right-inclined REE curve, flat HREE patterns, high 10,000 ∗ Ga/Al and intensively negative Eu anomalies, exhibiting an A2-type granite affinity with Y/Nb > 1.2 mostly. The primitive magma of these large quantities of granites was generated under a high temperature, low pressure, reduced and anhydrous environment indicating intense upwelling of asthenosphere. Combining with the positive uniform zircon eHf(t) values of −0.2 to +3.8 and decoupled eNd(t) values of −4.9 to −2.1 at t = 424 Ma, it can be concluded that subduction-related juvenile materials, probably calc-alkaline granitoids, are the source of these A-type granites. Geochemical studies of Wulonggou granites, spatial and temporal distributions of regional magmatism, metamorphism, and sedimentary records throughout the Eastern Kunlun Orogen Belt jointly indicate that the whole orogenic belt was in a typical post-collision extension setting and experienced an isostatic uplift during the middle Silurian triggered by delamination after the convergence of the northeastern margin of Gondwana.

The Eastern Kunlun Orogenic Belt lay on a significant tectonic boundary between Gondwana and Eurasia, produced widespread granitoids, and recorded at least two orogenic episodes since the Phanerozoic eon. In this paper, we provided new petrography, geochemistry, zircon U–Pb dating, and Lu–Hf isotopic research on the Middle–Late Silurian granitoids from the eastern segment of The Eastern Kunlun Orogenic Belt. These granitoids comprised of Zhongzhigou granodiorites and Xintuo alkali-feldspar granites, formed in the Middle–Late Silurian (420–425 Ma). Geochemically, the granodiorites exhibit high Sr/Y (31.2–35.2) as adakitic affinities, and alkali-feldspar granites belong to highly fractionated I-type granites. Additionally, granodiorites have zircon eHf(t) values show limited positive variations from − 0.5 to + 3.2, with two-stage Hf model ages (TDM2) of 1105–1306 Ma, while alkali-feldspar granites show negative zircon eHf(t) values ranged from − 4.6 to − 0.1 with two-stage Hf model ages (TDM2) of 1289–1541 Ma, implying that their primary magma originated from different sources. Taken together, we conclude that granodiorites had been derived from partial melting of the juvenile newly underplated mafic lower crust, while alkali-feldspar granites were derived from partial melting of the shallower ancient crustal source. Combined with temporal and spatial distribution of magmatic activities, sedimentation, and metamorphism, it can be concluded that the whole EKOB underwent a transition from syn-collision compression to post-collision extension and extensive crustal reworking occurred during the Middle–Late Silurian.

Angiotensin (Ang) A is formed by the decarboxylation of the N terminal residue of AngII. The present study determined whether this one amino acid change impacted effects of AngII on abdominal aortic aneurysm (AAA) formation in mice. Computational analyses implicated that AngA had comparable binding affinity to both AngII type 1 and 2 receptors as AngII. To compare effects of these two octapeptides in vivo, male low-density lipoprotein receptor (Ldlr) or apolipoprotein E (Apoe) deficient mice were infused with either AngII or AngA (1 μg/kg/min) for 4 weeks. While AngII infusion induced AAA consistently in both mouse strains, the equivalent infusion rate of AngA did not lead to AAA formation. We also determined whether co-infusion of AngA would influence AngII-induced aortic aneurysm formation in male Apoe−/− mice. Co-infusion of the same infusion rate of AngII and AngA did not change AngII-induced AAA formation. Since it was reported that a 10-fold higher concentration of AngA elicited comparable vasoconstrictive responses as AngII, we compared a 10-fold higher rate (10 μg/kg/min) of AngA infusion into male Apoe−/− mice with AngII (1 μg/kg/min). This rate of AngA led to abdominal aortic dilation in three of ten mice, but no aortic rupture, whereas the 10-fold lower rate of AngII infusion led to abdominal aortic dilation or rupture in eight of ten mice. In conclusion, AngA, despite only being one amino acid different from AngII, has diminished effects on aortic aneurysmal formation, implicating that the first amino acid of AngII has important pathophysiological functions.

The aim of the present study is to evaluate the vasodilation effects of Tongmai Yangxin Pills (TMYX) on rat mesenteric artery as well as its mechanism of action. The relaxation effects of TMYX extracts with different concentrations were determined on isolated rat mesenteric artery in normal condition as well as pretreating by phenylephrine and KCl. Vascular relaxation effects of TMTX were also determined in mesenteric artery preincubated with L-ANME and indomethacin or in endothelium denuded mesenteric artery. Moreover, effects of TMYX by 50 mg·L⁻¹ on NO secretion and the phosphorylation of eNOS in a cellular model of human umbilical vein endothelial cell (HUVEC) pretreated with or without L-NAME were also observed. The experimental results showed that TMYX has no obvious effect on vasodilation of arteries in normal or KCl pretreated condition, while it can dose-dependently relax the rat mesenteric artery with intact endothelium stimulated with phenylephrine at a maximal diastolic rate of (64.71±10.03)%. After preincubating with L-NAME for 15 min or removal of mesenteric artery endothelium, the maximal diastolic rate was decreased to (35.77±8.93)% and (25.85±10.84)% respectively. However, preincubating with indomethacin had no inhibitory effect on TMYX induced vascular relaxation. Meanwhile, TMYX at 50 mg·L⁻¹ could increase the expression of P-eNOS and the secretion of NO in HUVEC. L-NAME significantly inhibited NO release and phosphorylation of eNOS induced by TMYX. The results suggested TMYX exerted endothelium-dependent relaxation effects against PE-induced contractions of isolated rat mesenteric artery through NO-cGMP signaling pathway.

OBJECTIVE To study the effects of electroacupuncture (EA) in chronic constrictive injury (CCI) rat model and the expression of N-methyl-D-aspartate receptor type 2B (NR2B) in ipsilateral spinal dorsal horn in rats to explore the analgesic mechanisms of EA. METHODS According to the random number table, totally 180 rats were evenly divided into a sham group, a CCI group, and an EA group. CCI model was conducted with four 4-0 chromic gut ligatures loosely ligated around the left sciatic nerve 1 cm above the trifurcation. Rats in the EA group received 2 Hz EA therapy bilaterally at acupoints of Zusanli (ST 36) and Sanyinjiao (SP 6) once daily (30 min/d) for 30 days after surgery. Paw withdrawal thresholds (PWTs) were measured on 0 (baseline), 1, 3, 7, 15, 30 days after surgery. Rats were sacrificed on 0, 1, 3, 7, 15 and 30 days after surgery, and the L4-5 segments of spinal cord were removed to detect the expression of NR2B by immunohistochemistry and quantitative polymerase chain reaction. RESULTS PWTs in the CCI group were significantly lower than the sham group at Day 1-30 after surgery, and reached its lowest at Day 1 (P

The abnormal expression of microRNAs (miRNAs) in colorectal cancer (CRC) progression has been widely investigated. It was reported that the same hairpin RNA structure could generate mature products from each strand, termed 5p and 3p, which binds different target mRNAs. Here, we explored the expression, functions, and mechanisms of miR-514b-3p and miR-514b-5p in CRC cells and tissues. We found that miR-514b-3p was significantly down-regulated in CRC samples, and the ratio of miR-514b-3p/miR-514b-5p increased from advanced CRC, early CRC to matched normal colorectal tissues. Follow-up functional experiments illustrated that miR-514b-3p and miR-514b-5p had distinct effects through interacting with different target genes: MiR-514b-3p reduced CRC cell migration, invasion and drug resistance through increasing epithelial marker and decreasing mesenchymal marker expressions, conversely, miR-514b-5p exerted its pro-metastatic properties in CRC by promoting EMT progression. MiR-514b-3p overexpressing CRC cells developed tumors more slowly in mice compared with control cells, however, miR-514b-5p accelerated tumor metastasis. Overall, our data indicated that though miR-514b-3p and miR-514b-5p were transcribed from the same RNA hairpin, each microRNA has distinct effect on CRC metastasis.

Zinc finger protein 278 is a zinc finger transcription factor encoded on the 22q12.2 chromosome. Previous studies revealed that ZNF278 expression was significantly upregulated in colorectal cancer (CRC) tissue compared to adjacent non-tumor tissue. However, the expression and specific roles of ZNF278 in CRC remain unknown. ZNF278 expression was knocked down using specific siRNAs, which was confirmed by western blotting, and the effects of ZNF278 siRNAs on CRC cell proliferation were investigated. In addition, the effects of ZNF278 overexpression were confirmed by western blotting and cell proliferation assay. Correlations between ZNF278 and the ERK/MAPK pathway were also detected by western blotting. We found that ZNF278 knockdown significantly induced cell cycle arrest, resulting in cyclin D1/E1 downregulation and p21 upregulation. Moreover, we demonstrated that downregulation of ZNF278 decreased the proliferation of CRC cells via inhibition of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway for the first time. In conclusion, ZNF278 played a prominent role in the pathogenesis of CRC, and promoted CRC cell proliferation via the ERK/MAPK pathway, suggesting that it may act as a potential target in the diagnosis or treatment of CRC.

It is increasingly evident that long noncoding RNAs (lncRNA) have causative roles in carcinogenesis. In this study, we report findings implicating a novel lncRNA in gastric cancer, termed GAPLINC (gastric adenocarcinoma predictive long intergenic noncoding RNA), based on the use of global microarray and in situ hybridization (ISH) analyses to identify aberrantly expressed lncRNA in human gastric cancer specimens. GAPLINC is a 924-bp-long lncRNA that is highly expressed in gastric cancer tissues. GAPLINC suppression and with gene expression profiling in gastric cancer cells revealed alterations in cell migration pathways, with CD44 expression the most highly correlated. Manipulating GAPLINC expression altered CD44 mRNA abundance and the effects of GAPLINC on cell migration and proliferation were neutralized by suppressing CD44 expression. Mechanistic investigations revealed that GAPLINC regulates CD44 as a molecular decoy for miR211-3p, a microRNA that targets both CD44 and GAPLINC. Tissue ISH analysis suggested that GAPLINC overexpression defines a subgroup of patients with gastric cancer with very poor survival. Taken together, our results identify a noncoding regulatory pathway for the CD44 oncogene, shedding new light on the basis for gastric cancer cell invasiveness. Cancer Res; 74(23); 6890–902. ©2014 AACR.

Gastric cancer is the second after lung cause of cancer-related mortality in the world. Early detection and treatment can lead to a long survival time. Recently microarrays and next generation sequencing (NGS) have become very useful tools of comprehensive research into gastric cancer, facilitating the identification of treatment targets and personalized treatments. However, there are numerous challenges from cancer target discovery to practical clinical benefits. Although there are many biomarkers and target agents, only a minority of patients are tested and treated accordingly. Microarray technology with maturity was established more than 10 years ago, and has been widely used in the study of functional genomics, systems biology, and genomes in medicine. Second generation sequencing technology is more recent, but development is very fast, and it has been applied to the genome, including sequencing and epigenetics and many aspects of functional genomics. Here we review insights gained from these studies regarding the technology of microarray and NGS, how to elucidate the molecular basis of gastric cancer and identify potential therapeutic targets, and how to analyse candidate genes. We also discuss the challenges and future directions of such efforts.

Mafic–intermediate igneous rocks can provide important insights into the geodynamic evolution of orogenic belts. Here, we investigate the petrogenesis of mafic–intermediate intrusive igneous rocks in the East Kunlun Orogenic Belt (EKOB) to constrain regional geodynamic and magmatic evolution. These intrusive rocks are all located in the Permian-Triassic magmatic arc and parallel to the Paleo-Tethys ophiolite belt. Zircon U Pb dating results indicate that these various igneous rocks crystallized during the Triassic. The 250–248 Ma appinites display low SiO2 contents, high MgO contents, and marked enrichments in large-ion lithophile elements and depletions in high-field-strength elements; combined with their enriched eHf(t) values, these appinites were considered to be derived from lithospheric mantle modified by subducted slab-derived fluids. The Harizha host granodiorites and MMEs have indistinguishable crystallization ages of 235 Ma but distinct SiO2 contents and δ18OSMOW values, which, together with enriched Sr–Nd–Hf–Pb isotopes, suggest that their primary magmas were derived from the Mesoproterozoic lower crust (MLC) and subduction-modified lithospheric mantle (SMLM), respectively. The 241 Ma Baidungou quartz diorite and the 218 Ma Huanglonggou diorite porphyry show characteristics of medium SiO2 contents, moderate Mg# and eHf(t) values relative to contemporaneous mafic and granitic rocks; therefore they probably represent the mixing products of MLC-derived felsic and SMLM-derived mafic magmas. Different from the Baidungou diorite and Harizha granodiorite, the Huanglonggou diorite porphyries have adakitic characteristics, which suggests crustal thickening during the Late Triassic. A synthesis of these new data with previous researches shows that the EKOB undergone three magmatic peaks corresponding to three extension events (slab-rollback, slab-breakoff, and delamination) during the Triassic evolution.

In this paper, the dynamic stiffness method is used to research the dynamic stability characteristics of multi-span aluminum-based functionally graded material (FGM) fluid-conveying pipe enhanced by carbon nanotube. The influence of the different volume fraction on natural frequency, critical pressure and critical velocity of the FGM pipe is discussed. The influence of different radiuses on the natural frequency of the pipe and the influence of different span-ratios on the stability characteristics of the pipe are also discussed. It can be concluded that change of material component has significant influence on natural frequencies, critical velocity and critical pressure of the FGM pipe. The instability caused by flow velocity variation is different from that caused by pressure change. If the total length of the pipe remains unchanged, different span-ratios will have a great impact on the stability of the pipe. Increasing outside diameter of the pipe will cause the increase of the natural frequency.

Recent studies have increasingly linked microRNAs to colorectal cancer (CRC). MiR-194 has been reported deregulated in different tumor types, whereas the function of miR-194 in CRC largely remains unexplored. Here we investigated the biological effects, mechanisms and clinical significance of miR-194. Functional assay revealed that overexpression of miR-194 inhibited CRC cell viability and invasion in vitro and suppressed CRC xenograft tumor growth in vivo. Conversely, block of miR-194 in APCMin/+ mice promoted tumor growth. Furthermore, miR-194 reduced the expression of AKT2 both in vitro and in vivo. Clinically, the expression of miR-194 gradually decreased from 20 normal colorectal mucosa (N-N) cases through 40 colorectal adenomas (CRA) cases and then to 40 CRC cases, and was negatively correlated with AKT2 and pAKT2 expression. Furthermore, expression of miR-194 in stool samples was gradually decreased from 20 healthy cases, 20 CRA cases, then to 28 CRC cases. Low expression of miR-194 in CRC tissues was associated with large tumor size (P=0.006), lymph node metastasis (P=0.012) and shorter survival (HR =2.349, 95% CI = 1.242 to 4.442; P=0.009). In conclusion, our data indicated that miR-194 acted as a tumor suppressor in the colorectal carcinogenesis via targeting PDK1/AKT2/XIAP pathway, and could be a significant diagnostic and prognostic biomarker for CRC.

Zinc finger protein X-linked (ZFX) is a zinc finger transcription factor encoded on the X chromosome. Here, we found that ZFX expression was significantly upregulated in gastric cancer (GC) cell lines and tissues. Knockdown of ZFX induced significant apoptosis and cell cycle arrest in SGC7901 and MGC803 cells. Moreover, we demonstrated for the first time that knockdown of ZFX inhibited gastric cancer cell growth in vitro and in vivo via downregulating the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK-MAPK) pathway. Therefore, ZFX play a prominent role in GC tumorigenicity and may have potential applications in the diagnosis or treatment of GC.

Colorectal adenoma (CRA) is the precursor lesion of colorectal cancer (CRC). Several agents have been shown to be effective in the chemoprevention of CRA recurrence, but there has been little research on its primary prevention. Participants older than 50 years with no adenomas were recruited for our study and randomized to receive either 1 mg/day folic acid supplement or treatment without folic acid. After 3 years of follow-up, plasma folate and colonoscopy were evaluated. Seven hundred ninety-one participants (91.98%) completed the study. CRA occurred in 64 (14.88%) participants in the folic acid group and 132 (30.70%) in the control group [unadjusted risk ratio (RR), 0.49; 95% confidence interval (CI), 0.37–0.63; P < 0.01]; left-sided adenoma (unadjusted RR, 0.54; 95% CI, 0.38–0.76; P = 0.001) and advanced CRA (unadjusted RR, 0.36; 95% CI, 0.16–0.81; P = 0.01) were most common. There was no significance difference in the occurrence of three or more adenomas (unadjusted RR, 0.70; 95% CI, 0.36–1.77; P = 0.38) or right-sided adenoma (unadjusted RR, 0.55; 95% CI, 0.30–1.00; P = 0.07) between the two groups. Participants with low plasma folate may have a high risk of CRA. In conclusion, primary prevention with 1 mg/day folic acid supplementation could reduce the incidence of CRA, especially left-sided and advanced disease in those with no previous adenomas. People with differing baseline plasma folate levels should be given individualized treatment. Those with low plasma folate should be encouraged to take adequate supplements; plasma folate should be elevated to an effective therapeutic level, which may reduce the incidence of CRA. Cancer Prev Res; 6(7); 744–52. ©2013 AACR.

The polycomb group protein enhancer of zeste homologue 2 (EZH2), which has histone methyltransferase (HMT) activity, is overexpressed in malignant tumours. However, the role of EZH2 in colorectal cancer (CRC) invasion is little known. Here we investigated the clinical significance, biological effects, and mechanisms of EZH2 signalling. Knockdown of EZH2 significantly reduced cell invasion and secretion of matrix metalloproteinases 2/9 (MMP2/9) in in vitro studies. Knockdown of EZH2 dramatically increased overall survival and decreased metastasis of lung in in vivo studies. Conversely, overexpression of EZH2 significantly increased lung metastasis and shortened overall survival when compared with control tumours. EZH2-induced CRC cell invasion may depend on down-regulation of vitamin D receptor (VDR), which is considered to be a marker of CRC invasion. EZH2 regulates the histone trimethylation of lysine 27 (H3K27me3) in the VDR promoter. Moreover, we found that STAT3 directly binds to the EZH2 promoter and regulates VDR down-regulation in CRC cells. Significant inverse correlations were observed between the expression of EZH2 and pSTAT3 and that of VDR in CRC tissues compared with normal tissue in patients. We show the role of EZH2 in CRC metastasis and identify VDR as a target gene of EZH2. EZH2 expression may be directly regulated by STAT3, and STAT3 may play an important role in EZH2-mediated VDR down-regulation in CRC. This pathway may provide potential targets in aggressive CRC.

The extracellular signal-regulated kinase (ERK), mitogen-activated protein kinase (MAPK) pathway is an important cell proliferation pathway. We previously reported that the transport protein particle complex 4 (TRAPPC4), ERK2 interaction may activate ERK1/2, modulate pERK2 nuclear localization and regulate proliferation and apoptosis in colorectal cancer (CRC) cells. The present study further investigated the function of the TRAPPC4–ERK2 interaction in CRC in vitro and in vivo. Silencing of TRAPPC4 induced G0/G1 phase cell cycle arrest, upregulated p21 and downregulated cyclin B1 in CRC cells. Overexpression of TRAPPC4 after ERK2 silencing decreased the percentage of G0/G1 phase cells, increased the percentage of G2/M and S phase cells, downregulated p21, upregulated cyclin B1, and enhanced CRC cell viability. Immunohistochemical staining revealed that knockdown of TRAPPC4 downregulated pERK2, whereas overexpression of TRAPPC4 upregulated pERK2. Epidermal growth factor (EGF) stimulated upregulation of TRAPPC4 and pERK2 in SW1116 cells; EGF stimulation or overexpression of TRAPPC4 induced pERK2 nuclear translocation. Silencing of TRAPPC4 reduced SW1116 xenograft tumor growth in vivo, whereas overexpression of TRAPPC4 increased tumor growth, compared to control tumors. Moreover, modulation of TRAPPC4 expression in vivo affected the levels of pERK2 in the cytoplasm and nucleus and expression of p21. These results conclusively demonstrate that TRAPPC4 regulates ERK2 activation and also affects the distribution of activated pERK2 in CRC cells. The ability of ERK2 to play a role in colorectal carcinogenesis depends, at least in part, on TRAPPC4. © 2013 Wiley Periodicals, Inc.

The gene, collagen triple helix repeat containing 1 (CTHRC1), has been reported to increase in several kinds of human solid cancers and is associated with tumor invasion and metastasis. To date, the expression and function of CTHRC1 in gastric cancer (GC) have not been reported. The aim of this study was to investigate the expression levels and regulatory transcription mechanisms of CTHRC1 in GC. Immunohistochemical analysis revealed that CTHRC1 expression was markedly increased in carcinoma compared with normal gastric mucosa, chronic atrophic gastritis, and intestinal metaplasia (P

Although the two isoforms of signal transducer and activator of transcription 5 (STAT5) protein, STAT5a and STAT5b, have 94% sequence identity, they are encoded by different genes. Previous studies have been unable to define clearly the roles of the STAT5 genes in colorectal cancer (CRC). To investigate the role of STAT5 isoforms in CRC oncogenesis, immunohistochemical staining was performed. Colorectal adenocarcinomas showed higher expression of STAT5a/5b than normal colonic mucosa (P

Abnormalities in signal transducer and activator of transcription (STAT) signaling, especially STAT3 and STAT5, are involved in the oncogenesis of several human cancers, including gastric cancer (GC). However, the downstream targets of STAT3 and STAT5 are not fully identified, and the precise roles and the prognostic value of STAT3 and STAT5 in GC have not been fully characterized. In this study, we used ChIP-on-chip to identify STAT3 and STAT5 target genes on a whole genome scale in AGS cells, a human GC cell line. A total of 2,514 and 1,314 genes were identified as STAT3 and STAT5 target genes, which were mainly related to cell growth, metabolism, differentiation, adhesion, immune response, and stress response. Furthermore, we depleted STAT3 and STAT5 with a small interfering RNA, respectively. Our results demonstrate that STAT3, but not STAT5, is involved in GC cell growth and cell cycle progression through regulation of gene expression, such as Bcl-2, p16(ink4a) and p21(waf1/cip1). Moreover, expression of pSTAT3(Tyr705) correlates with TNM stage, differentiation and survival, and is a significant prognostic factor in GC. Therefore, our findings provide novel evidence that STAT3 may be a potential therapeutic target for GC treatment and pSTAT3(Tyr705) expression can predict prognosis in GC.

Aberrant janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling is involved in the oncogenesis of several cancers. Suppressors of cytokine signaling (SOCS) genes and SH2-containing protein tyrosine phosphatase 1 (SHP1) proteins, which are negative regulators of JAK/STAT signaling, have been reported to have tumor suppressor functions. However, in colorectal cancer (CRC) cells, the mechanisms that regulate SOCS and SHP1 genes, and the cause of abnormalities in the JAK/STAT signaling pathway, remain largely unknown. The present study shows that trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, leads to the hyperacetylation of histones associated with the SOCS1 and SOCS3 promoters, but not the SHP1 promoter in CRC cells. This indicates that histone modifications are involved in the regulation of SOCS1 and SOCS3. Moreover, upregulation of SOCS1 and SOCS3 expression was achieved using TSA, which also significantly downregulated JAK2/STAT3 signaling in CRC cells. We also demonstrate that TSA suppresses the growth of CRC cells, and induces G1 cell cycle arrest and apoptosis through the regulation of downstream targets of JAK2/STAT3 signaling, including Bcl-2, survivin and p16(ink4a) . Therefore, our data demonstrate that TSA may induce SOCS1 and SOCS3 expression by inducing histone modifications and consequently inhibits JAK2/STAT3 signaling in CRC cells. These results also establish a mechanistic link between the inhibition of JAK2/STAT3 signaling and the anticancer action of TSA in CRC cells.

AML1-ETO, a fusion protein generated by the chromosomal translocation t(8;21), is frequently associated with acute myeloid leukemia (AML). In addition to blocking differentiation, AML1-ETO is also shown to induce growth arrest in AML cells, which is unfavorable for leukemogenesis harboring the t(8;21) translocation. However, its precise mechanism is still unclear. Here we provide the first demonstration that the conditional expression of AML1-ETO by the ecdysone-inducible system dramatically increases the expression of connexin 43 (CX43), together with growth arrest at G1 phase in leukemic U937 cells. We also show that the CX43 induction inhibits the proliferation of U937 cells at G1 phase, while the suppression of CX43 expression by small interfering RNA (siRNA) effectively overcomes the growth-inhibitory effect of AML1-ETO in leukemic cells. Furthermore, either AML1-ETO or CX43 induction elevates cell-cycle negative regulator P27kip1 protein by inhibiting its degradation, which is antagonized by siRNA against CX43. Taken together, our data indicate that CX43 plays a role in AML1-ETO-induced growth arrest possibly through the accumulation of P27kip1 protein. The potential mutation or/and epigenetic alterations of CX43 and its related gene(s) deserve to be explored in AML1-ETO-positive AML patients. J. Cell. Physiol. 208: 594–601, 2006. © 2006 Wiley-Liss, Inc.

To investigate the association of childhood hemophagocytic syndrome (HPS) with human parvovirus B19 (HPVB19) infection, and to analyze the clinical features of this disease.ELISA and quantitative real-time PCR were used to detect HPVB19-IgM, HPVB19-IgG and HPVB19-DNA in 65 children with HPS (HPS group) and 65 healthy children (control group). The HPS group was divided into HPVB19-infected (n=14) and non-infected (n=51) groups according to the detection results of HPVB19-DNA. The clinical data of two groups were compared.The positive rate of HPVB19-IgM in the HPS group (26%, 17/65) was significantly higher than that in the control group (9%, 6/65) (P=0.011), and there was no significant difference in the positive rate of HPVB19-IgG between the HPS (38%, 25/65) and control groups (29%, 19/65) (P=0.266). The infection rate of HPVB19 in the HPS group (22%, 14/65) was significantly higher than that in the control group (3%, 2/65) (P=0.001). Compared with the non-infected group, the HPVB19-infected group had significantly lower platelet count and hemoglobin level on admission, significantly more severe liver function damage, a significantly earlier onset time, and a significantly longer course of disease (P0.05).The pathogenesis of HPS may be associated with HPVBl9 infection. HPVBl9-infected children with HPS have more acute onset, more severe clinical manifestations, and a longer disease duration.

Purpose: The role and clinical implication of the transmembrane protein with EGF and two follistatin motifs 2 (TMEFF2) in gastric cancer is poorly understood. Experimental Design: Gene expression profile analyses were performed and Gene Set Enrichment Analysis (GSEA) was used to explore its gene signatures. AGS and MKN45 cells were transfected with TMEFF2 or control plasmids and analyzed for gene expression patterns, proliferation, and apoptosis. TMEFF2 expression was knocked down with shRNAs, and the effects on genome stability were assessed. Interactions between TMEFF2 and SHP-1 were determined by mass spectrometry and immunoprecipitation assays. Results: Integrated analysis revealed that TMEFF2 expression was significantly decreased in gastric cancer cases and its expression was negatively correlated with the poor pathologic stage, large tumor size, and poor prognosis. GSEA in The Cancer Genome Atlas (TCGA) and Jilin datasets revealed that cell proliferation, apoptosis, and DNA damage–related genes were enriched in TMEFF2 lower expression patients. Gain of TMEFF2 function decreased cell proliferation by increasing of apoptosis and blocking of cell cycle in gastric cancer cells. The protein tyrosine phosphatase SHP-1 was identified as a binding partner of TMEEF2 and mediator of TMEFF2 function. TMEFF2 expression positively correlated with SHP-1, and a favorable prognosis was more likely in patients with gastric cancer with higher levels of both TMEFF2 and SHP-1. Conclusion: TMEFF2 acts as a tumor suppressor in gastric cancer through direct interaction with SHP-1 and can be a potential biomarker of carcinogenesis. Clin Cancer Res; 20(17); 4689–704. ©2014 AACR.

In this study we investigated the biological role and mechanism of miR-198 in colorectal carcinoma (CRC). MiR-198 expression was shown to exhibit a strongly negative correlation with lymph node invasion, distant metastasis and patient survival in examinations of colorectal cancer tissues and paired normal colorectal mucosa tissues. fucosyl transferase 8 (FUT8) was identified as a potential target of miR-198 in bioinformatics analysis and luciferase reporter assays. Overexpression of miR-198 in CRC cell lines decreased FUT8 levels as shown by immunofluorescence analysis, and inhibited cell proliferation, migration, and invasion. These anti-tumor phenotypes were rescued by reconstitution of FUT8 expression. Furthermore, miR-198 was shown to target the 3'UTR of FUT8 directly to downregulate FUT8 expression at both mRNA and protein levels in qRT-PCR and Western blot analyses, respectively. In vivo, restoration of miR-198 significantly inhibited xenograft growth and invasion of CRC tumors in nude mice. Therefore, it could be concluded that miR-198 suppresses the proliferation and invasion of CRC by directly targeting FUT8.

To compare the outcomes and potential morbidities of laparoscopic spleen-preserving distal pancreatectomy with or without splenic vessel preservation in patients with benign or low-grade malignant pancreatic lesions.Twenty patients who underwent spleen-preserving distal pancreatectomy were retrospectively analyzed. All the patients had benign or low-grade malignant pancreatic lesions that had not invaded the spleen. Twelve patients underwent Kimura's procedure and eight patients underwent Warshaw's. Perioperative data, and procedure-specific complications were compared between the two groups.Age, gender, and body mass index were comparable between the two groups. Operative time and intraoperative blood loss were significantly lower for patients who underwent Warshaw's procedure than for those who received Kimura's (p0.05 for both). There were no significant differences between the two groups with regard to perioperative blood transfusions, length of postoperative hospital stays, or complication rates. Splenic infarction and gastric varices developed only in patients who underwent Warshaw's procedure (one case each).Our results suggest that the Kimura technique should be the first choice for patients with benign or low-grade malignant pancreatic lesions. Warshaw's technique was associated with a higher incidence of several complications. However, Warshaw's can increase the success rate of splenic preservation in some cases.