Analysis on EZH2: mechanism identification of related CeRNA and its immunoassay in hepatocellular carcinoma.

OBJECTIVE: To screen the possible potential signaling pathways related to enhancer of zeste homolog 2 (EZH2) based on ceRNA mechanism, and to analyze the correlation between E2H2 and depths of various immune cell infiltration depths. The relationship between different immune checkpoints were also analyzed. METHODS: First, the expression of EZH2 in pan-cancer (18 malignancies) was analyzed with the TCGA database. Hepatocellular carcinoma (HCC) tissues of 374 cases and normal tissues of 50 cases were analyzed in terms of the differential expression, overall survival (OS) and progression-free-survival (PFS). Then, we conducted GO and KEGG enrichment analysis on target gene. We also analyzed mRNA-miRNA and MicroRNA (miRNA)- long non-coding RNA (lncRNA) correlation with starbase databse, so as to determine the potential ceRNA mechanism associated with EZH2. Finally, immunoassay and drug-sensitivity analysis of EZH2 was performed. RESULTS: Seven potential EZH2-related ceRNA pathways were screened out, namely lncRNA: Small Nucleolar RNA Host Gene 1 (SNHG1), SNHG 3, and SNHG 6-miR-101-3p-EZH2; and lncRNA: Long Intergenic Non-Protein Coding RNA 1978 (LINC01978), SNHG12, Ring Finger Protein 216 Pseudogene 1 (RNF216P1), and Coiled-coil Domain Containing 18 Antisense RNA 1 (CCDC18-AS1)-let-7c-5p-EZH2. Finally, 4 potential EZH2-related ceRNA pathways were identified through qPCR.According to immune correlation analysis, EZH2 may be positively correlated with T cells follicular helper, T cells Cluster of differentiation (CD)4 memory activated, Macrophages M0, and B cells memory (P < 0.05, cof > 0.2); while be negatively correlated with T cells CD4 + memory resting (P < 0.05, cof < -0.2). And EZH2 is positively correlated with Programmed Cell Death 1 (PDCD1) (R = 0.22), CD274 (R = 0.3) and Cytotoxic T-Lymphocyte Associated Protein 4 (CTLA4) (R = 0.23). According to drug sensitivity analysis, patients in the high expression group were more susceptible to the effects of various drugs including Sorafenib, 5-Fluorouracil, Doxorubicin, Etoposide, Paclitaxel, and Vinorelbine than those with low expression. CONCLUSION: This study revealed seven potential pathways of Enhancer of Zeste Homolog 2 (EZH2)-related ceRNA mechanisms: lncRNA (SNHG3, 6) -Mir-101-3P-ezh2; lncRNA (SNHG12, RNF216P1)-let-7c-5p-EZH2. We also analyzed the immunity and drug sensitivity of EZH2. Our study proves that EZH2 still has great research prospects in HCC.

Pan-cancer analysis of SYNGR2 with a focus on clinical implications and immune landscape in liver hepatocellular carcinoma.

BACKGROUND: Synaptogyrin-2 (SYNGR2), as a member of synaptogyrin gene family, is overexpressed in several types of cancer. However, the role of SYNGR2 in pan-cancer is largely unexplored. METHODS: From the TCGA and GEO databases, we obtained bulk transcriptomes, and clinical information. We examined the expression patterns, prognostic values, and diagnostic value of SYNGR2 in pan-cancer, and investigated the relationship of SYNGR2 expression with tumor mutation burden (TMB), microsatellite instability (MSI), immune infiltration, and immune checkpoint (ICP) genes. The gene set enrichment analysis (GSEA) software was used to perform pathway analysis. Besides, we built a nomogram of liver hepatocellular carcinoma patients (LIHC) and validated its prediction accuracy. RESULTS: SYNGR2 was highly expressed in most cancers. The high expression of SYNGR2 significantly reduced the overall survival (OS), disease-specific survival (DSS), disease-free interval (DFI), and progression-free interval (PFI) in multiple types of cancer. Also, receiver operating characteristic (ROC) curve analysis demonstrated that SYNGR2 showed high accuracy in distinguishing cancerous tissues from normal ones. Moreover, SYNGR2 expression was correlated with TMB, MSI, immune scores, and immune cell infiltrations. We also analyzed the association of SYNGR2 with immunotherapy response in LIHC. Finally, a nomogram including SYNGR2 and pathologic T, N, M stage was built and exhibited good predictive power for the OS, DSS, and PFI of LIHC patients. CONCLUSION: Overall, SYNGR2 is a critical oncogene in various tumors. SYNGR2 participates in the carcinogenic progression, and may contribute to the immune infiltration in tumor microenvironment. Our study suggests that SYNGR2 can serve as a predictor related to prognosis in pan-cancer, especially LIHC.

Degradation of soybean meal proteins by wheat malt endopeptidase and the antioxidant capacity of the enzymolytic products.

This study investigated the hydrolysis effect of the endopeptidase from wheat malt on the soybean meal proteins. The results indicated that the endopeptidase broke the peptide bonds of soybean meal proteins and converted the alcohol- and alkali-soluble proteins into water-soluble and salt-soluble proteins. In addition, wheat malt endopeptidase did not break the disulfide bonds between proteins but affected the conformation of disulfide bonds between substrate protein molecules, which were changed from the gauche-gauche-trans (g-g-t) vibrational mode to the trans-gauche-trans (t-g-t) vibrational mode. Wheat malt endopeptidase exhibited the highest enzymatic activity at 2 h of enzymatic digestion, demonstrating the fastest hydrolytic rate of soybean meal proteins. Compared with the samples before enzymatic hydrolysis, the total alcohol- and alkali-soluble proteins were decreased by 11.89% but the water- and salt-soluble proteins were increased by 11.99%, indicating the hydrolytic effect of endopeptidase. The corresponding water-soluble proteins had molecular weights of 66.4-97.2, 29-44.3, and 20.1 kDa, while the salt-soluble proteins had molecular weights of 44.3-66.4, 29-44.3, and 20.1 kDa, respectively. The degree of enzymatic hydrolysis of soybean meal reached the maximum at 8 h. The newly created proteins exhibited significantly antioxidant properties, which were inversely related to the molecular weight. Proteins with molecular weight <3 kDa had the highest antioxidant performance with an antioxidant capacity of 1.72 ± 0.03 mM, hydroxyl radical scavenging rate of 98.04%, and ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] radical scavenging capacity of 0.44 ± 0.04 mM.

[Grain oil quality formation and metabolism-related genes difference expression of Paeonia suffruticosa cv. 'Fengdan' grown at different altitudes.] / 不同海拔'凤丹'ç‰¡ä¸¹ç±½ç²’æ²¹è„‚å“è´¨å½¢æˆåŠä»£è°¢ç›¸å ³åŸºå› å·®å¼‚è¡¨è¾¾.

We measured the morphological index, nutritional composition and the expression analysis of key genes during grain development of Paeonia suffruticosa cv. 'Fengdan' grown at altitudes of 100, 650 and 1010 m in Luo-yang. The aim of this study was to examine differences in grain yield traits and the transformation of soluble sugar, starch, soluble protein and fatty acid contents, as well as the related enzyme activity and differential expression of key genes in oil metabolism. The results showed that grain yield traits increased with altitudes and that the growth period of grain at the higher altitudes was longer than that at low and mid altitudes. The soluble sugar and starch in mature grains increased with altitudes, while soluble protein and crude fat did not change. During grain development, the activities of sucrose synthase (SS) and sucrose phosphate synthase (SPS) first decreased and then increased, with the lowest occurred at 90 d after flowering. The activities of pyruvate dehydrogenase (PDH), glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminease (GOT) increased rapidly during 50-90 d after flowering and peaked at 90 d. The relative expression of acetyl-CoA carboxylase (ACCase) and stearoyl-ACP desaturase (SAD) peaked at 50 d after flowering, and ω-6 fatty acid desaturase 2 (FAD2) peaked at 90 d, in oil tree peony grain at different altitudes. There was a negative correlation of soluble sugar and starch with the accumulation of soluble protein and crude fat. SPS activity was positively correlated with the contents of soluble sugar and starch, and negatively correlated with the contents of soluble protein and crude fat during grain development. Activities of GPT and GOT were negatively associated with the content of soluble sugar and the content of starch, and had a highly significant positive correlation with the contents of soluble protein and crude fat. Activity of PDH was positively correlated with the content of soluble proteins and activities of GPT and GOT, and negatively correlated with the contents of soluble sugar and starch. It suggested that nutrient accumulation in the process of grain development of tree peony was transformed from sugar to crude fat and protein, and that metabolic enzymes, such as SPS, PDH, GPT and GOT, played an important role in this process. Palmitate acid, stearic acid and linoleic acid were negatively correlated with the relative increment of α-linolenic acid, indicating that fatty acid desaturation process in the grain development of tree peony was towards the direction of α-linolenic acid synthesis. The relative expression of ACCase, SAD, and FAD2 was positively correlated with the relative increment of α-linolenic acid accumulation, which played an important role in α-linolenic acid synthesis. The oil quality of tree peony grain was relatively stable at different altitudes, but grain production increased with altitude. Planting oil tree peony at mid to high altitudes could be an important strategy for the efficient use of marginal land in Luoyang.

Bioinformatic-based mechanism identification of E2F1-related ceRNA and E2F1 immunoassays in hepatocellular carcinoma.

Background: E2F1 is an important transcription factor. Previous studies have shown that the overexpression of E2F1 is closely related to the occurrence and development of hepatocellular carcinoma (HCC). However, the current research on the regulatory mechanism of E2F1 is still insufficient. This study sought to identify valuable therapeutic E2F1-related targets for HCC. Methods: HCC-related transcriptome data and patient clinical information downloaded from The Cancer Genome Atlas (TCGA) database. The expression of the E2F1 gene in pan-cancer was analyzed using the Tumor IMmune Estimation Resource (TIMER) 2.0 database, and the expression level of E2F1 in HCC was verified using the Gene Expression Profiling Interactive Analysis database. The overall survival (OS) and progression-free survival (PFS) in HCC patients were also analyzed. Subsequently, based on the Encyclopedia of RNA Interactomes (ENCORI) database, we adopted E2F1 as the research objective and identified the target long non-coding RNAs (lncRNAs) and microRNAs that suggested the competing endogenous RNA (ceRNA) mechanisms related to E2F1. We also performed a correlation analysis of E2F1 using the R language package that contained immune cell and immune checkpoint information. Finally, the drug sensitivity of E2F1 was detected using the R language package, "pRRophetic." Results: Ultimately, the following 6 potential ceRNA-based pathways targeting E2F1 were identified-lncRNA: LINC01224, PCBP1-AS1, and ITGA9-AS1-miR-29b-3p-E2F1; lncRNA: SNHG7 and THUMPD3-AS1, and LINC02323-miR-29c-3p-E2F1. Cluster of differentiation (CD)4 memory activated T cells, memory B cells, eosinophils, and T follicular helper cells were positively correlated with E2F1 (P<0.05), and monocytes, naïve B cells, and CD4 memory resting T cells were negatively correlated with E2F1 (P<0.05). The immune checkpoint analysis showed that E2F1 was positively correlated with PDCD1, CTLA4, and LAG3 (P>0.2). According to the drug sensitivity analysis, E2F1 may be sensitive to 39 drugs (P<0.05). Conclusions: This study provides a valuable direction for researching transcription factor E2F1, which may be conducive in identifying research targets for HCC-related molecular biological therapy and immunotherapy in future.

Dynamic Contrast-Enhanced Ultrasonography with Sonazoid for Diagnosis of Microvascular Invasion in Hepatocellular Carcinoma.

The aim of the present study was to investigate the imaging features observed in pre-operative Sonazoid contrast-enhanced ultrasound (SZ-CEUS) and the correlations with the presence of microvascular invasion (MVI) in hepatocellular carcinoma (HCC) patients. In this single-center retrospective study, 31 patients with surgically and histopathologically confirmed HCC lesions were included. Patients were classified according to the presence of MVI into the MVI-positive group (n = 15) and MVI-negative group (n = 16). The CEUS examinations were performed within 2 or 3 d before surgery. Features, including tumor necrosis and ultrasound contrast agent (UCA) distribution characteristics in the arterial phase (AP), tumor types (single nodular [SN] or non-single nodular [non-SN]) in the post-vascular phase (PVP), wash-in time, wash-in slope, time to peak (TTP) and peak intensity (PI), were assessed. Univariate analysis revealed statistically significant differences between the two groups with respect to tumor necrosis (p = 0.002), inhomogeneous distribution of contrast agent in the AP (p = 0.001) and non-SN type in the PVP (p < 0.001). There was no significant difference in the quantitative parameters. Multivariate analysis revealed that non-SN type in the PVP was a significant independent risk factor for MVI of HCC (odds ratio = 30.51, 95% confidence interval [CI]: 2.335-398.731, p = 0.009). The area under the receiver operating characteristic (ROC) curve (AUC), sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 0.873, 93.3%, 81.3%, 82.4% and 92.9%, respectively. Thus, SZ-CEUS can provide useful information for the diagnosis of MVI in HCC.

NEU4 inhibits motility of HCC cells by cleaving sialic acids on CD44.

Hepatocellular carcinoma (HCC) is an extremely metastatic tumor. Sialic acids (SAs) are associated with cancer development and metastasis. NEU4 is a sialidase that removes SAs from glycoconjugates, while the function of the NEU4 in HCC has not been clearly explored. In our research, we found the NEU4 expression was significantly down-regulated in HCC tissues, which was correlated with high grades and poor outcomes of HCC. The NEU4 expression could be regulated by histone acetylation. In the functional analysis of NEU4, the cell motility was inhibited when NEU4 was overexpressed, and restored when NEU4 expression was down-regulated. Similarly, NEU4 over-expressed HCC cells showed less metastasis in athymic nude mice. Further study revealed that NEU4 could inhibit cell migration by enzymatic decomposition of SAs. Our results verified a NEU4 active site (NEU4E235) and overexpressing inactivates NEU4E235A that weakens the inhibition ability to cell migration. Further, 70 kinds of specific interacting proteins of NEU4 including CD44 were identified through mass spectrum. Moreover, the α2,3-linked SAs on CD44 were decreased and the hyaluronic acid (HA) binding ability was increased when NEU4 over-expressed or activated. Additionally, the mutation of CD44 with six N-glycosylation sites showed less sensibility to NEU4 on cell migration compared with wild-type CD44. In summary, our results revealed the mechanism of low expression of NEU4 in HCC and its inhibitory effect on cell migration by removal of SAs on CD44, which may provide new treatment strategies to control the motility and metastasis of HCC.

CPEB3-mediated MTDH mRNA translational suppression restrains hepatocellular carcinoma progression.

Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is a sequence-specific RNA-binding protein. We had reported that CPEB3 is involved in hepatocellular carcinoma (HCC) progression. However, the underlying mechanisms of CPEB3 in HCC remain unclear. In this study, we firstly performed RNA immunoprecipitation to uncover the transcriptome-wide CPEB3-bound mRNAs (CPEB3 binder) in HCC. Bioinformatic analysis indicates that CPEB3 binders are closely related to cancer progression, especially HCC metastasis. Further studies confirmed that metadherin (MTDH) is a direct target of CPEB3. CPEB3 can suppress the translation of MTDH mRNA in vivo and in vitro. Besides, luciferase assay demonstrated that CPEB3 interacted with 3'-untranslated region of MTDH mRNA and inhibited its translation. Subsequently, CPEB3 inhibited the epithelial-mesenchymal transition and metastasis of HCC cells through post-transcriptional regulation of MTDH. In addition, cpeb3 knockout mice are more susceptible to carcinogen-induced hepatocarcinogenesis and subsequent lung metastasis. Our results also indicated that CPEB3 was a good prognosis marker, which is downregulated in HCC tissue. In conclusion, our results demonstrated that CPEB3 played an important role in HCC progression and targeting CPEB3-mediated mRNA translation might be a favorable therapeutic approach.

Novel Metabolomics Serum Biomarkers for Pancreatic Ductal Adenocarcinoma by the Comparison of Pre-, Postoperative and Normal Samples.

Background: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive human malignancies. The metabolomic approaches are developed to discover the novel biomarkers of PDAC. Methods: 550 preoperative, postoperative PDAC and normal controls (NCs) serums were employed to characterize metabolic alterations in training and validation sets by LC-MS. Results: The results of PLS-DA analysis indicated that three groups could be distinguished clearly and the post-PDAC group is adjacent to a normal group as compared with pre-PDAC group. Further results showed that histidinyl-lysine significantly increased whereas docosahexaenoic acid and LysoPC (14:0) decreased in pre-PDAC patients as compared with NCs. And these three markers had a significant tendency to recover after tumor resection. The validation set results revealed that for CA19-9 negative patients, 92.3% (12/13) of them can be screened using these three metabolites. The combination of these markers could significantly improve the diagnostic performance for PDAC, with higher sensitivity (0.93), specificity (0.92) and AUC (0.97). Moreover, network and pathways analyses explored the latent relationship among differential metabolites. The glycerolipid metabolism and primary bile acid synthesis showed variation in network and pathway analysis. Conclusions: These three markers combined with CA199 displayed high sensitivity and specificity for detecting PDAC patients from NCs. The results indicated that these three metabolites could be regarded as potential biomarkers to distinguish PDAC from NCs.

Extracellular vesicles from human urine-derived stem cells inhibit glucocorticoid-induced osteonecrosis of the femoral head by transporting and releasing pro-angiogenic DMBT1 and anti-apoptotic TIMP1.

Osteonecrosis of the femoral head (ONFH) frequently occurs after glucocorticoid (GC) treatment. Extracellular vesicles (EVs) are important nano-sized paracrine mediators of intercellular crosstalk. This study aimed to determine whether EVs from human urine-derived stem cells (USC-EVs) could protect against GC-induced ONFH and focused on the impacts of USC-EVs on angiogenesis and apoptosis to explore the mechanism by which USC-EVs attenuated GC-induced ONFH. The results in vivo showed that the intravenous administration of USC-EVs at the early stage of GC exposure could rescue angiogenesis impairment, reduce apoptosis of trabecular bone and marrow cells, prevent trabecular bone destruction and improve bone microarchitecture in the femoral heads of rats. In vitro, USC-EVs reversed the GC-induced suppression of endothelial angiogenesis and activation of apoptosis. Deleted in malignant brain tumors 1 (DMBT1) and tissue inhibitor of metalloproteinases 1 (TIMP1) proteins were enriched in USC-EVs and essential for the USC-EVs-induced pro-angiogenic and anti-apoptotic effects in GC-treated cells, respectively. Knockdown of TIMP1 attenuated the protective effects of USC-EVs against GC-induced ONFH. Our study suggests that USC-EVs are a promising nano-sized agent for the prevention of GC-induced ONFH by delivering pro-angiogenic DMBT1 and anti-apoptotic TIMP1. STATEMENT OF SIGNIFICANCE: This study demonstrates that the intravenous injection of extracellular vesicles from human urine-derived stem cells (USC-EVs) at the early stage of glucocorticoid (GC) exposure efficiently protects the rats from the GC-induced osteonecrosis of the femoral head (ONFH). Moreover, this study identifies that the promotion of angiogenesis and inhibition of apoptosis by transferring pro-angiogenic DMBT1 and anti-apoptotic TIMP1 proteins contribute importantly to the USC-EVs-induced protective effects against GC-induced ONFH. This study suggests the promising prospect of USC-EVs as a new nano-sized agent for protecting against GC-induced ONFH, and the potential of DMBT1 and TIMP1 as the molecular targets for further augmenting the protective function of USC-EVs.

An Often Easily Missed Injury in the Presence of Orthopaedic Trauma: A Case Series of Derived Injury.

Derived disaster is a common concept in emergencies such as earthquakes. With the progress of society, the incidence of fractures caused by high-energy trauma has increased year by year. After the first injury, the possibility of derived injury caused by the original injury also increases rapidly. Orthopaedic surgeons, especially trauma orthopaedic surgeons, lack sufficient understanding and recognition of this kind of injury. The purpose of this article is to present a case series of an often missed injury pattern that is associated with an original injury. The diagnosis of derived injury may go unrecognized in a considerable number of cases and delayed treatment decreases the success rate of soft tissue repairing; therefore, a high index of suspicion and a proper early diagnosis is of paramount importance. We also describe the current surgical management used by the authors, and propose the concept of "derived injury", studying its clinical significance in traumatic orthopaedics.

Onset of double-diffusive convection in near-critical gas mixtures.

Near the thermodynamic critical point, the physical properties of binary fluids exhibit large variations in response to small temperature and concentration differences, whose effects on the onset of double-diffusive convection are reported here. The vertical symmetry is broken, irregular penetrative instability occurs, and cat's eye patterns are identified in the fingering regime and oscillatory regime, respectively. A new parameter Θ is defined which indicates how the variations of physical properties influence flow fields. It is seen through numerical simulations that the Boussinesq approximation with constant physical properties has limited applicability, and that the Boussinesq equations with variable properties and density will describe all features seen. This conclusion is based on comparisons with the fully compressible, variable-property system.

Metabolic profiling analysis upon acylcarnitines in tissues of hepatocellular carcinoma revealed the inhibited carnitine shuttle system caused by the downregulated carnitine palmitoyltransferase 2.

The carnitine shuttle system (CSS) plays a crucial role in the transportation of fatty acyls during fatty acid ß-oxidation for energy supplementation, especially in cases of high energy demand, such as in cancer. In this study, to systematically characterize alterations of the CSS in hepatocellular carcinoma (HCC), acylcarnitine metabolic profiling was carried out on 80 pairs of HCC tissues and adjacent noncancerous tissues (ANTs) by using ultra-performance liquid chromatography coupled to mass spectrometry. Twenty-four acylcarnitines classified into five categories were identified and characterized between HCCs and ANTs. Notably, increased saturated long-chain acylcarnitines (LCACs) and decreased short- and medium-chain acylcarnitines (S/MCACs) were simultaneously observed in HCC samples. Subsequent correlation network and heatmap analysis indicated low correlations between LCACs and S/MCACs. The mRNA and protein expressions of carnitine palmitoyltransferase 2 (CPT2) was significantly downregulated in HCC samples, whereas CPT1A expression was not significantly changed. Correspondingly, the relative levels of S/MCACs were reduced and those of LCACs were increased in BEL-7402/CPT2-knockdown cells compared to negative controls. Both results suggested that decreased shuttling efficiency in HCC might be associated with downregulation of CPT2. In addition, decreases in the mRNA expression of acetyl-CoA acyltransferase 2 were also observed in HCC tissues and BEL-7402/CPT2-knockdown cells, suggesting potential low ß-oxidation efficiency, which was consistent with the increased expression of stearoyl-CoA desaturase 1 in both samples. The systematic strategy applied in our study illustrated decreased shuttling efficiency of the carnitine shuttle system in HCC and can provide biologists with an in-depth understanding of ß-oxidation in HCC.

Climate change risk perception in global: Correlation with petroleum and liver disease: A meta-analysis.

BACKGROUND: Liver diseases have been bound to environmental factors, inclusive of air pollution. The exposure of workers to petrochemicals counts as a possible cause of Liver diseases, whereas results are inconsistent with the previous studies. In this study, a meta-analysis is conducted to assess the pooled risk. METHODS AND FINDING: A systematic search was performed by related researchers. Correlations are analyzed among petroleum and liver cirrhosis mortality, fatty liver, alanine amino transferase (abbreviated as ALT), aspartate amino transferase (abbreviated as AST). Pooled risk ratios (RR) with 95% confidence interval (CI) and effect size(ES) with 95% confidence interval are calculated. Sensitivity analysis and publication bias are also tested. Data are analyzed from 5 studies involving 296 participants. Results are incorporated through adopting a random effects meta-analysis. Working in a petrochemical plant shall not increase the death risk posed by cirrhosis (RR = 0.44, 95% CI [0.36; 0.54]). Yet the incidence of fatty liver increases (RR = 1.22, 95% CI [1.21; 1.23]). Abnormal incidence of ALT and AST also increases. CONCLUSIONS: Occupational exposure plays an important role in causing ALT abnormalities and fatty liver among oil workers, but not a risk factor of cirrhosis, AST abnormalities and liver cancer.

CADM2, as a new target of miR-10b, promotes tumor metastasis through FAK/AKT pathway in hepatocellular carcinoma.

BACKGROUND: Cell adhesion molecules (CADMs) comprise of a protein family whose functions include maintenance of cell polarity and tumor suppression. Hypo-expression of CADM2 gene expression has been observed in several cancers including hepatocellular carcinoma (HCC). However, the role and mechanisms of CADM2 in HCC remain unclear. METHODS: The expression of CADM2 and miRNA-10b (miR-10b) in HCC tissues and cell lines were detected using real-time PCR and Western blotting. Immunofluorescence was used to detect Epithelial-mesenchymal transition (EMT) progression in HCC cell lines. Dual-luciferase reporter assay was used to determine miR-10b binding to CADM2 3'UTR. Wound healing assay and Transwell assay were performed to examine the migration and invasion of HCC cells. RESULTS: We report the effect of CADM2 as a tumor suppressor in HCC. Firstly, we confirmed that CADM2 expression was significantly down regulated in HCC tissues compared to normal tissues according to TCGA data analysis and fresh HCC sample detection. Secondly, overexpression of CADM2 could inhibit EMT process, migratory and invasion ability of HCC cells. Furthermore, the results indicated that CADM2 is a direct target of miR-10b in HCC cells and miR-10b/CADM2 modulates EMT process and migration ability via focal adhesion kinase (FAK) /AKT signaling pathway in HCC. CONCLUSIONS: Our study demonstrates that miR-10b-CADM2-FAK/AKT axis plays an important role in HCC metastasis, which might be a novel potential therapeutic option for HCC treatment.

Raltitrexed Inhibits HepG2 Cell Proliferation via G0/G1 Cell Cycle Arrest.

Raltitrexed (RTX) is an antimetabolite drug used as a chemotherapeutic agent for treating colorectal cancer, malignant mesothelioma, and gastric cancer. The antitumor capacity of RTX is attributed to its inhibitory activity on thymidylate synthase (TS), a key enzyme in the synthesis of DNA precursors. The current study is aimed at investigating the potential antitumor effects of RTX in liver cancer. Using the HepG2 cell line as an in vitro model of liver cancer, we evaluated the effects of RTX on cell proliferation employing both a WST-8 assay and a clone formation efficiency assay. In addition, we monitored the ultrastructure changes of HepG2 cells in response to RTX with transmission electric microscopy. To investigate the mechanism underlying the regulation of cell proliferation by RTX, we analyzed cell cycle using cell flow cytometry. Moreover, real-time PCR and Western blot analyses were conducted to examine expression levels of cell cycle regulatory proteins cyclin A and cyclin-dependent kinase 2 (CDK2), as well as their mediators tumor suppressor genes p53 and p16. Our results demonstrate that RTX inhibits HepG2 proliferation by arresting the cell cycle at G0/G1. This cell cycle arrest function was mediated via downregulation of cyclin A and CDK2. The observed elevated expression of p53 and p16 by RTX may contribute to the reduction of cyclin A/CDK2. Our study indicates that RTX could serve as a potential chemotherapeutic agent in the treatment of hepatocellular carcinoma.

Identification of drug-target interaction from interactome network with 'guilt-by-association' principle and topology features.

MOTIVATION: Identifying drug-target protein interaction is a crucial step in the process of drug research and development. Wet-lab experiment are laborious, time-consuming and expensive. Hence, there is a strong demand for the development of a novel theoretical method to identify potential interaction between drug and target protein. RESULTS: We use all known proteins and drugs to construct a nodes- and edges-weighted biological relevant interactome network. On the basis of the 'guilt-by-association' principle, novel network topology features are proposed to characterize interaction pairs and random forest algorithm is employed to identify potential drug-protein interaction. Accuracy of 92.53% derived from the 10-fold cross-validation is about 10% higher than that of the existing method. We identify 2272 potential drug-target interactions, some of which are associated with diseases, such as Torg-Winchester syndrome and rhabdomyosarcoma. The proposed method can not only accurately predict the interaction between drug molecule and target protein, but also help disease treatment and drug discovery. CONTACTS: zhanchao8052@gmail.com or ceszxy@mail.sysu.edu.cn SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Mid-term Outcomes of Anterior Cervical Fusion for Cervical Spondylosis With Sympathetic Symptoms.

STUDY DESIGN: Prospective study. OBJECTIVE: The purpose of this study is to elucidate mid-term outcomes of anterior cervical fusion for cervical spondylosis with sympathetic symptoms (CSSS). SUMMARY OF BACKGROUND DATE: The terminology, pathogenesis, diagnosis, and treatment of CSSS remain controversial. Surgical treatment of CSSS has been rarely reported. This is the first prospective study to evaluate the mid-term outcome of surgical treatment of CSSS. METHODS: Thirty-one patients who were diagnosed with CSSS in 2006 were evaluated prospectively. All patients were assigned to undergo anterior cervical fusion with posterior longitudinal ligament (PLL) resection and followed up for ≥5 years. Sympathetic symptoms such as vertigo, headache, and tinnitus, etc. were evaluated using the sympathetic symptom 20-point score. Neurological status was assessed using the Japanese Orthopedic Association (JOA) score. Clinical and radiologic data were prospectively collected before surgery, and at 1 week, 2 months, 6 months, 2 years, and 5 years after surgery. Surgical complications and morbidities of other diseases during the follow-up were also recorded. RESULTS: The mean 20-point score decreased significantly from 7.3±3.5 before surgery to 2.2±2.7 at the final follow-up (P<0.001), giving a mean recovery rate of 66.1%±50.3%. Good to excellent results were attained in 80.6% of these patients. The sympathetic symptoms were relieved in 23 of the 31 patients in the early postoperative period, and 5 patients in 2 months. No relief of sympathetic symptoms was found in 3 patients. The mean JOA score improved significantly from 12.0±1.9 before surgery to 14.8±1.5 by the end of the follow-up (P<0.001). No late neurological deterioration was found in this group. CONCLUSIONS: The mid-term outcomes of anterior cervical fusion with PLL resection for CSSS have been satisfactory. Differential diagnosis before surgery is of great importance. PLL may play a role in presenting sympathetic symptoms.

Specific N-glycans of Hepatocellular Carcinoma Cell Surface and the Abnormal Increase of Core-α-1, 6-fucosylated Triantennary Glycan via N-acetylglucosaminyltransferases-IVa Regulation.

Glycosylation alterations of cell surface proteins are often observed during the progression of malignancies. The specific cell surface N-glycans were profiled in hepatocellular carcinoma (HCC) with clinical tissues (88 tumor and adjacent normal tissues) and the corresponding serum samples of HCC patients. The level of core-α-1,6-fucosylated triantennary glycan (NA3Fb) increased both on the cell surface and in the serum samples of HCC patients (p < 0.01). Additionally, the change of NA3Fb was not influenced by Hepatitis B virus (HBV)and cirrhosis. Furthermore, the mRNA and protein expression of N-acetylglucosaminyltransferase IVa (GnT-IVa), which was related to the synthesis of the NA3Fb, was substantially increased in HCC tissues. Knockdown of GnT-IVa leads to a decreased level of NA3Fb and decreased ability of invasion and migration in HCC cells. NA3Fb can be regarded as a specific cell surface N-glycan of HCC. The high expression of GnT-IVa is the cause of the abnormal increase of NA3Fb on the HCC cell surface, which regulates cell migration. This study demonstrated the specific N-glycans of the cell surface and the mechanisms of altered glycoform related with HCC. These findings lead to better understanding of the function of glycan and glycosyltransferase in the tumorigenesis, progression and metastasis of HCC.

Downregulation of long noncoding RNA MALAT1 induces epithelial-to-mesenchymal transition via the PI3K-AKT pathway in breast cancer.

The metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) regulates cell motility via the transcriptional or post-transcriptional control of motility-related genes. Whether MALAT1 plays a critical role in cancer progression in breast cancer remains unclear. In this study, we found that MALAT1 was downregulated in breast tumor cell lines and cancer tissue, and showed that knockdown of MALAT1 in breast cancer cell lines induced an epithelial-to-mesenchymal transition (EMT) program via phosphatidylinositide-3 kinase-AKT pathways. Furthermore, lower expression of MALAT1 in breast cancer patients was associated with shorter relapse-free survival. Thus, our results indicate for the first time that MALAT1 is a novel regulator of EMT in breast cancer and may be a potential therapeutic target for breast cancer metastasis.