Search

With the aim to improve the hygroscopic stability of palmatine chloride (PMTCl), an unreported single crystal of PMT—SSA (C21H20NO4·C7H5O6S·H2O) is prepared by the solvent evaporation method and characterized. A comprehensive analysis of the structure and Hirshfeld surface reveals that π—π stacking plays a crucial role in accumulating PMT—SSA. As expected, the hygroscopic stability of PMT—SSA is significantly improved compared with that of PMTCl. This may be due to the presence of π—π stacking in the structure of PMT—SSA which improves the hygroscopic stability. The present work inaugurates an avenue for improving the hygroscopic stability of active pharmaceutical ingredients and provides a novel crystalline product with a bright commercial prospect. Unfortunately, the maximum solubility of PMT—SSA is lower than that of PMTCl. How to improve its solubility will be explored in subsequent studies. However, it is surprising that PMT—SSA shows advantages with regard to antibacterial properties in comparison to PMTCl.

The FGF/FGFR signaling axis deregulation of the fibroblast growth factor receptor (FGFR) family is closely related to tumorigenesis, tumor progression and drug resistance to anticancer therapy. And fibroblast growth factor receptor 3 (FGFR3) is one member of this family. In this study, we aimed to investigate the effect of siRNA-induced knockdown of FGFR3 on the biological behaviors of intrahepatic cholangiocarcinoma (ICC). The expression levels of FGFR3 were determined in three intrahepatic cholangiocarcinoma cell lines RBE, HUCCT1 and HCCC9810 cell lines by Western blot. FGFR3 expression in RBE cell line was knocked down by siRNA. Our study found that knockdown of FGFR3 inhibited the migration, invasion and proliferation of ICC cells using Wound healing assay, Transwell migration and invasion assays and Cell proliferation assay. And significantly down-regulated the protein expression levels of MMP2, cyclinD1, and NCadherin, but had no significant effect on MMP9, cyclinD3, vimentin, E-cadherin protein. In addition, we found that ERK/c-Myc presumably is its signaling pathway by bioinformatics analysis and Western blot verification. To sum up, knockdown of FGFR3 inhibited the migration, invasion and proliferation of ICC cells. It demonstrated that FGFR3 probably becomes a therapeutic target for ICC and increases the proportion of potentially curable intrahepatic cholangiocarcinoma patients treated with FGFR inhibitors.

Background: Our previous studies showed that tetraspanin CD151 was implicated in the progression of hepatocellular carcinoma (HCC), mainly depending on the formation of functional complexes with molecular partners, including Mortalin. In this study, we investigate the role of mortalin in CD151-depedent progression of HCCs. Methods: Immunofluorescent staining, western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to investigate the expression and location of CD151 and Mortalin in four HCC cell lines with different metastatic ability. The relationship between Mortalin and CD151 was investigated in HCCLM3 cells using co-immunoprecipitation. CD151 or Mortalin expression in HCC cells were modified by transfection technology. Wound-healing assay and Transwell assay were used to assay the role of CD151 and Mortalin in cell migration and invasion. The expression and prognostic implication of CD151 and Mortalin in 187 cases of HCCs were analyzed. Results: Expression of Mortalin in HCC cells was positive related to their metastatic ability and its tendency was in line with the expression of CD151. Immunofluorescent staining showed that Mortalin was located in cytoplasm, while positive staining for CD151 was observed in cytoplasm and membrane of HCC cells. co-IP revealed that Mortalin formed a complex with CD151. Down-regulation of Mortalin induced a moderate decreased CD151 protein, but not CD151 mRNA, while inhibition of CD151 did not influence the expression of Mortalin at the level of both protein and mRNA. Interference of Mortalin significantly inhibited the invasion and migration of HCC cells with high CD151 expression and partially restored the invasion and migration of HCC cells induced by CD151 over-expression. Clinically, high Mortalin expression correlated with malignant phenotype of HCC, such as microvascular invasion (p=0.017) and tumor diameter (p=0.001). HCC patients expressing high Mortalin were tend to have higher expression of CD151. HCC patients expressing high level of CD151 showed the poorer prognosis in a Mortalin-dependent manner. Conclusions: Mortalin maybe stabilize of the structure of CD151-dependent tetraspanin-enriched microdomains and implicate in the progression of HCC.

Social network data publishing is dynamic, and attackers can perform association attacks based on social network directed graph data at different times. The existing social network privacy protection technology has low performance in dealing with large-scale dynamic social network directed graph data, and anonymous data publishing does not meet the needs of community structure analysis. A Dynamic Social Network Directed Graph K-In&Out-Degree Anonymity (DSNDG-KIODA) method to protect community structure is proposed. The method is based on the dynamic grouping anonymity rule to anonymize the dynamic K-in&out-degree sequence, and the virtual node distribution is added in parallel to construct an anonymous graph. The node information is transmitted based on the GraphX, and the virtual node pairs are selected and deleted according to the change of the directed graph modularity to reduce information loss. The experimental results show that the DSNDG-KIODA method improves the efficiency of processing large-scale dynamic social network directed graph data, and ensures the availability of community structure analysis when data is released.

Intrahepatic cholangiocarcinoma (ICC) can invade and metastasize. EIF5A2 is involved in the invasive metastatic process of several digestive malignancies. However, its role in ICC is yet to be elucidated.Immunohistochemistry (IHC) and Western blot (WB) were used to detect the level of EIF5A2 in the tumor specimens of ICC patients and evaluate the correlation between its expression and clinicopathological characteristics. The significance of EIF5A2 in the prognosis of ICC patients was further evaluated by Kaplan-Meier and Cox regression analysis. In addition, CCK-8, EdU, Transwell invasion, and scratch assays were utilized to detect tumor cell proliferation, invasion, and metastasis. Furthermore, the role of EIF5A2 in ICC cells was evaluated after modification of EIF5A2 expression.The level of EIF5A2 protein was significantly higher in ICC than in adjacent tissues. This high expression in the tumor samples was significantly associated with malignant phenotypes, such as lymph node metastasis (LNM), microvascular or bile duct invasion, and poor differentiation. ICC patients with high expression of EIF5A2 had short overall survival and a high cumulative recurrence rate. The multifactorial analysis showed that EIF5A2 is an independent prognostic marker. Furthermore, high levels of EIF5A2 may activate the PI3K/AKT/mTOR signaling pathway and upregulate Cyclin D1, Cyclin D3, MMP2, and MMP9 to promote ICC cell proliferation, migration, and invasion.The current study found that EIF5A2 promotes ICC progression and is a prognostic biomarker and candidate therapeutic target for ICC patients.

Background: The present study aimed to investigate the prognostic value of the neutrophil-to-lymphocyte ratio (NLR) in distal cholangiocarcinoma (DCC) following radical surgery. Methods: The clinicopathological data of 59 patients with DCC were retrospectively reviewed. Patients were treated by radical surgery and diagnosed by postoperative pathology at the Second Affiliated Hospital of Kunming Medical University (Yunnan, China), between July 2015 and December 2017. The optimal cut-off value for the NLR was determined by generating receiver operating characteristic (ROC) curves. Kaplan–Meier survival analysis and Cox proportional hazards models were used to determine the risk factors and independent risk factors influencing the prognosis of patients with DCC. Results: According to the ROC curve, the optimal cut-off value for the NLR was 2.933. The results of Kaplan–Meier survival analysis and the Cox proportional hazards model showed that carbohydrate antigen 125, NLR, perineural, vascular and fat invasion, regional lymph node metastasis, and the American Joint Committee on Cancer stage were risk factors for DCC; the only independent risk factor to affect the prognosis of DCC patients was the NLR. Conclusions: The preoperative NLR plays an important guiding role in evaluating the prognosis of patients with DCC, and an increase in the NLR is associated with poor patient prognosis.

Background/Aims: Cardiac remodeling is a critical pathogenetic process leading to heart failure. Suppressor of cytokine signaling-3 (SOCS3) is demonstrated as a key negative regulator of the gp130 receptor to inhibit cardiac hypertrophy. However, the role of SOCS3 in deoxycorticosterone-acetate (DOCA)-salt-induced cardiac remodeling remains unclear. Methods: Cardiac-specific SOCS3 knockout (SOCS3cKO) and wild-type (WT) C57BL/6J mice were subjected to uninephrectomy and DOCA-salt for 3 weeks. The effect of SOCS3 on cardiac remodeling and inflammation was evaluated by histological analysis. Gene and protein levels were measured by real-time PCR and immunoblotting analysis. Results: After DOCA-salt treatment, the expression of SOCS3, activation of gp130/JAK/STAT3, cardiac dysfunction and fibrosis in DOCA-salt mice were significantly elevated, which were markedly attenuated by eplerenone, a specific mineralocorticoid receptor (MR) blocker. Moreover, DOCA-salt-induced cardiac dysfunction, hypertrophy, fibrosis and inflammation were aggravated in SOCS3cKO mice, but were significantly reduced in AAV9-SOCS3-injected mice. These effects were mostly associated with activation of gp130/STAT3/AKT/ERK1/2, TGF-β/Smad2/3 and NF-κB signaling pathways. Conclusions: Our data demonstrate that loss of SOCS3 in cardiomyocytes promotes DOCA-salt-induced cardiac remodeling and inflammation, and it may be a novel potential therapeutic target for hypertensive heart disease.

Recent reports show that B7-H4 is highly expressed in a variety of tumor cells, functions as a negative regulator of T cells and then promotes tumor progression. However, its expression and role in intrahepatic cholangiocarcinoma (ICC) remain unclear. In present study, B7-H4 expression in ICC and peritumoral tissues was determined at the level of mRNA and protein, and its bioactivity in ICC cells was studied after modification of B7-H4 expression. Then, the mechanism related to tumor progression induced by B7-H4 expression in ICC cells was explored. Finally, clinical significance of B7-H4 expression in ICC patients was further analyzed. The results showed that B7-H4 expression in ICC was much higher than that in peritumoral tissues at the level of both mRNA and protein. The high level of B7-H4 in ICC cells induced epithelial-to-mesenchymal transitions and promoted invasion and metastasis of tumor cells through activation of ERK1/2 signaling. The elevated B7-H4 expression was associated with the downregulated Bax, upregulated Bcl-2 expression, and activation of caspase-3. Clinically, high B7-H4 expression in tumor samples was significantly related to malignant phenotype, such as lymph node metastasis, high tumor stage, and poor differentiation. ICC patients with high expression of B7-H4 had shorter overall survival (OS) and disease-free survival. Moreover, the B7-H4 expression was an independent prognostic factor for predicting OS and tumor recurrence of ICC patients after operation. In conclusion, high expression of B7-H4 promotes tumor progression of ICC and may be a novel therapeutic target for ICC patients.

Objective Although several statistical methods for adjusting for missing data have been developed and are widely applied in research, few studies have investigated these methods in adjusting for missingness in datasets that aim to estimate the prevalence of dementia. We attempted to develop a more feasible approach for handling missingness in a cross-sectional study among elderly. Methods Five methods of estimating prevalence, including stratified weighting (SW), inverse-probability weighting (IPW), hot deck imputation (HDI), ordinal logistic regression (OLR) and multiple imputation (MI), were applied to handle the missing data yielded by a dataset that include 2231 non-responders. Results Compared with the results of the complete case analysis, the differences in the prevalence rates of dementia and mild cognitive impairment (MCI) calculated by the prevalence-estimating methods after adjusting for non-responders were less than 7% and 6%, respectively. In contrast to the results of other methods, the estimated prevalence of dementia and MCI calculated by MI increased when more predictive factors were included, and the lowest rate of missing data was achieved using MI. Using the participants’ ages, the cognitive screening sores and activity of daily life sores as predictive variables when correcting for missingness induced relatively larger effects on the estimated dementia prevalence. Conclusions When adjusting for missingness while estimating the prevalence of dementia in cross-sectional studies, a simple method, such as SW, is recommended when limited information is available, whereas MI is the preferred method when additional information is available. Further simulation studies are needed to determine the optimal approach.

The immunoproteasome is a multicatalytic protease complex in all eukaryotic cells, which plays a key role in regulating essential cellular processes. However, the role of immunoproteasome subunit β2i in regulation of cardiac fibrosis and inflammation in deoxycorticosterone-acetate (DOCA)/salt mice remains unknown. Wild-type (WT) and β2i knockout (KO) mice were subjected to uninephrectomy and DOCA/salt treatment for 21 days. Blood pressure was measured by the tail-cuff system. Cardiac function and remodeling were examined by echocardiography, hematoxylin-eosin (H&E) and Masson's trichrome staining. The gene and protein expressions were detected by quantitative real-time PCR, and Western blot analysis. After 21 days, DOCA/salt treatment significantly up-regulated the expression of β2i mRNA and protein in the hearts. Moreover, systolic blood pressure and heart weight/body weight (HW/BW) ratio were significantly higher in DOCA/salt mice than in sham groups, and these effects were markedly reversed in β2i knockout mice. Importantly, DOCA/salt-induced cardiac fibrosis, inflammation and the expression of collagen I, collagen III, α-SMA, IL-1β, IL-6 and TNF-α in the wild-type hearts, which were markedly attenuated by β2i knockout. These beneficial effects were due, at least in part, to the inhibition of IκBα/NF-κB and TGF-β1/Smad2/3 signaling pathways. Collectively, these findings indicate that knockout of β2i ameliorates DOCA/salt-induced cardiac fibrosis and inflammation, and may be a novel potential therapeutic target for hypertensive heart diseases.

The present study aimed to investigate the effects of intestinal endotoxemia (IETM) in a rat model of aluminum neurotoxicity established by D-galactose and aluminum trichloride (AlCl3). Adult Wistar rats were administered D‑galactose and AlCl3 to create the aluminum neurotoxicity model. The learning and memory abilities of the rats were subsequently observed using a Morris water maze test and the serum levels of lipopolysaccharide (LPS), tumor necrosis factor (TNF)‑α, interleukin (IL)‑1, diamine oxidase (DAO), glutamine (Gln) and glutaminase were measured. The expression of S‑100β in the serum was detected using an enzyme‑linked immunosorbent assay. The expression levels of the amyloid β‑protein (Aβ) precursor (APP), presenilin 1 (PS1), β‑site APP‑cleaving enzyme (BACE), zona occludens protein (ZO)‑1 and Aβ 1‑40 in the brain of rats were detected via reverse‑transcription polymerase chain reaction, western blotting and immunohistochemistry. The levels of LPS, TNF‑α, IL‑1, DAO, Gln and S‑100β in serum and the mRNA and protein expression levels of APP, PS1, BACE and Aβ1‑40 in the brain were markedly increased in the model rats compared with controls. The level of glutaminase in the serum and the expression of ZO‑1 in the brain were decreased in the model rats compared with controls. IETM was present in the rat model of aluminum neurotoxicity established by D‑galactose and AlCl3 and may be important in the development of this neurotoxicity.

Emerging evidence has shown that leptin, an adipocyte-derived cytokine that is closely associated with obesity, play a significant role in carcinogenesis and tumorigenesis. However, its impact on gallbladder cancer (GBC) remains unclear. In this study, we firstly found that leptin and its functional receptor OB-Rb were significantly co-expressed in human GBC tissues and cell lines, the content of which were higher than those in normal human gallbladder tissues. Treatment with leptin promoted the proliferation, migration and invasion of GBC cells, which were attenuated by OB-Rb shRNA. Blocking in the G2/M period of cell cycle, increasing of MMP3 and MMP9, increasing of VEGF-C/D, activation of SOCS3/JAK2/p-STAT3 pathway was demonstrated after treatment with leptin. All of these positive responses were attenuated by OB-Rb receptor shRNA. Taken together, our findings suggest that leptin promoted the proliferation, migration and invasion of GBC cells by increasing OB-Rb expression through the SOCS3/JAK2/p-STAT3 signal pathway. Targeting the leptin/OB-Rb axis could be an attractive therapeutic strategy for treatment of GBC.

Background: The aim of this study was to assess the serum zinc levels in patients with common oral mucosal diseases by comparing these to healthy controls. Material and Methods: A total of 368 patients, which consisted of 156 recurrent aphthous stomatitis (RAS) patients, 57 oral lichen planus (OLP) patients, 55 burning mouth syndrome (BMS) patients, 54 atrophic glossitis (AG) patients, 46 xerostomia patients, and 115 sex-and age-matched healthy control subjects were enrolled in this study. Serum zinc levels were measured in all participants. Statistical analysis was performed using a one-way ANOVA, t-test, and Chi-square test. Results: The mean serum zinc level in the healthy control group was significantly higher than the levels of all other groups ( p < 0.001). No individual in the healthy control group had a serum zinc level less than the minimum normal value. However, up to 24.7% (13/54) of patients with AG presented with zinc deficiency, while 21.2% (33/156) of patients with RAS, 16.4% (9/55) of patients with BMS, 15.2% (7/46) of patients with xerostomia, and 14.0% (8/57) of patients with OLP were zinc deficient. Altogether, the zinc deficiency rate was 19.02% (70/368) in the oral mucosal diseases (OMD) group (all patients with OMD). The difference between the OMD and healthy control group was significant ( p < 0.001). Gender differences in serum zinc levels were also present, although not statistically significant. Conclusions: Zinc deficiency may be involved in the pathogenesis of common oral mucosal diseases. Zinc supplementation may be a useful treatment for oral mucosal diseases, but this requires further investigation; the optimal serum level of zinc, for the prevention and treatment of oral mucosal diseases, remains to be determined.

Aims Nucleotide-binding oligomerization domain-containing protein 2 (NOD2), an intracellular pattern recognition receptor, which plays an important role in the innate immunity and inflammation. However, its role in myocardial ischemia/reperfusion (I/R) injury remains unknown. In this study, we sought to determine the role of NOD2 on cardiac I/R injury. Main methods Mice were induced 30 min ischemia followed by 24 h of reperfusion. Histological examinations were performed on heart sections with Evans blue and triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin (H&E) staining, immunohistochemistry and immunofluorescence staining. The messenger RNA (mRNA) expression and protein levels were detected by real-time polymerase chain reaction (RT-PCR) and western blot analysis respectively. Key findings I/R injury markedly upregulated NOD2 expression in heart tissue. Treatment of WT mice with NOD2 ligand (MDP) significantly increased infarct size, the number of apoptotic cells and inflammatory cells, as compared with wild-type mice after I/R injury. Furthermore, MDP enhanced I/R-induced cardiomyocyte apoptosis and inflammation in vitro, and these effects were attenuated by NOD2-siRNA. The mechanism of NOD2 on cardiac I/R injury is partly associated with JNK, p38MAPK and NF-κB signaling pathways. Significance NOD2 aggravates myocardial I/R injury by inducing cardiomyocyte apoptosis and inflammation through JNK, p38MAPK and NF-κB signaling pathways. This study provides insight into better understanding the molecular mechanism of NOD2, which may be served as a potential target for the treatment of myocardial I/R injury.

Our previous studies revealed that tetraspanin CD151 plays multiple roles in the progression of hepatocellular carcinoma (HCC) by forming a functional complex with integrin α6β1. Herein, we generated a monoclonal antibody (mAb) that dissociates the CD151/integrin α6β1 complex, and we evaluated its bioactivity in HCCs. A murine mAb, tetraspanin CD151 (IgG1, called CD151 mAb 9B), was successfully generated against the CD151-integrin α6β1 binding site of CD151 extracellular domains. Co-immunoprecipitation using CD151 mAb 9B followed by Western blotting detected a 28 kDa protein. Both immunofluorescent and immunohistochemical staining showed a good reactivity of CD151 mAb 9B in the plasma membrane and cytoplasm of HCC cells, as well as in liver cells. In vitro assays demonstrated that CD151 mAb 9B could inhibit neoangiogenesis and both the mobility and the invasiveness of HCC cells. An in vivo assay showed that CD151 mAb 9B inhibited tumor growth potential and HCC cells metastasis. We successfully produced a CD151 mAb 9B targeting the CD151/integrin α6β1-binding domain, which not only can displayed good reactivity to the CD151 antigen but also prevented tumor progression in HCC.

Patent ductus arteriosus is the third most common congenital heart disease and resulted from the persistence of ductal patency after birth. Ductus arteriosus closure involves functional and structural remodeling, controlled by many factors. The changes in plasma protein levels associated with PDA closure are not known. Here we for the first time demonstrate six key differential plasma proteins in human patent ductus arteriosus patients using proteomic technology and present a model to illustrate the constriction and closure of ductus arteriosus. Differentially expressed proteins were analyzed by using isobaric tags for relative and absolute quantification and validated by enzyme-linked immunosorbent assay in new samples. The proteomic data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD008568. We found 74 upregulated and 98 downregulated proteins in the plasma of patients with PDA. Five decreased proteins (platelet factor 4, fibrinogen, von Willebrand factor, collagen, and mannose binding lectin-associated serine protease-2) and one increased protein (fibronectin) may increase the risk of patent ductus arteriosus. Those proteins are closely related to platelet activation and coagulation cascades, complement mannan-binding-lectin, and other systemic signaling pathways. Our findings for the first time indicate that the differential proteins involved in different pathways may play key roles in the nonclosure of the ductus arteriosus in humans and may be developed as biomarkers for diagnosis. All those findings may be served as the basis of understanding the etiology and pathogenesis of patent ductus arteriosus.

The incidence of intrahepatic cholangiocarcinoma (ICC) is progressively increasing worldwide, and its prognosis remains poor. Accumulating evidence has demonstrated that tumor necrosis factor receptor-associated factor 4 (TRAF4), an adaptor protein, is involved in the carcinogenesis and progression of several tumor types. However, the function of TRAF4 in predicting prognosis, and mediating migration and invasion of ICC remains to be elucidated. In the present study, immunohistochemistry, western blotting and reverse transcription-quantitative polymerase chain reaction assays were used to determine that the expression of TRAF4 at the mRNA and protein levels in ICC tissues was significantly higher compared with that in non‑tumor tissues. The overexpression of TRAF4 was positively correlated with poor differentiation, regional lymphatic metastasis, and high tumor‑node-metastasis staging. Inhibiting the expression of TRAF4 using small interfering RNA decreased the migration and invasion of ICC cells in vitro. In addition, the AKT inhibitor perifosine eliminated the effect of TRAF4 on the invasion and migration of ICC cells in vitro. Clinically, the overexpression of TRAF4 was correlated with shorter overall survival rate and elevated recurrence rate in patients with ICC. Furthermore, patients with ICC with a high expression of TRAF4 and lymphatic metastasis were closely associated with a poorer prognosis compared with the other groups. Multivariate analysis indicated that the overexpression of TRAF4 was an independent prognostic indicator for patients with ICC. It was identified that a high level of TRAF4 facilitated the invasiveness of ICC cells via the activation of AKT signaling. The overexpression of TRAF4 may be a prognostic biomarker and candidate therapeutic target for patients with ICC.

Rheumatoid arthritis (RA) severely affects the quality life of patients due to its high association with disability. Traditional Chinese medicines have been reported to exert notable therapeutic effects on RA. The Chinese medicinal prescription Wang‑Bi Tablet (WB) has been successfully used to clinically treat RA for many years; however, its pharmacological mechanism of action is largely unclear. In the present study, adjuvant‑induced arthritis (AIA) rats were used to evaluate the anti‑inflammatory effects of WB and western blotting was used to explore the molecular mechanisms. The experimental results demonstrated that WB treatment significantly reduced arthritis score and hind‑paw volume. Furthermore, synovial hyperplasia, inflammatory cell infiltration and joint destruction were ameliorated by WB. The expression levels of the proinflammatory cytokines interleukin (IL)‑1β, tumor necrosis factor‑α and IL‑6, were reduced in the joints of WB‑treated rats. Western blotting revealed that WB could also inhibit excessive activation of nuclear factor (NF)‑κB and Janus kinase (JAK)‑signal transducer and activator of transcription 3 (STAT3) signaling pathways. These results indicated that the therapeutic effects of WB on AIA may be accomplished through inhibition of the NF‑κB and JAK‑STAT3 signaling pathways. These findings provide experimental evidence to support WB as a therapeutic agent for the treatment of patients with RA.

Feeding muscles of the buccal mass of Aplysia are innervated by cholinergic and serotonergic neurons. Buccal muscle 15 contracts in response to acetylcholine (ACh). During feeding arousal, ACh-elicited contraction of muscle 15 is potentiated by serotonin (5-HT). This paper demonstrates a dissociated cell preparation of muscle 15 in which cellular mechanisms regulating contraction can be investigated. Dissociated muscle fibers contracted in response to both KCl and ACh. Serotonin (10-6 M) significantly potentiated the shortening caused by both KCl and ACh. Potentiation lasted at least 4 min, similar to potentiation in intact muscles. Four types of currents recorded by patch clamp methods are illustrated. With 540 mM KCl in the patch electrode, stretch-activated channels having a chord conductance of 150 pS are observed in on-cell patches. In whole cell configuration, ACh elicits inward current at a holding potential of -60 mV. With high potassium in the electrode, depolarization elicits an outward current. The voltage-dependent outward current is blocked with cesium in the electrode and 4-aminopyridine and tetraethylammonium outside the cells. The remaining voltage-dependent inward current is calcium dependent. The voltage-dependent inward and outward currents are activated within the range of depolarization produced by ACh and may therefore play roles in regulating contractile responses elicited by ACh.

Background The aim of this study was to evaluate the association between hematinic deficiencies and recurrent aphthous stomatitis (RAS). Material and Methods 517 RAS patients and 187 healthy controls were enrolled in the present study. Hematinic deficiencies, including serum ferritin, folic acid, and vitamin B12 deficiencies were assessed for each participant. Gender and age were taken into account and the collected data were statistically analysed. Results Compared with the healthy controls, a significantly higher overall frequency of hematinic deficiencies was found in RAS patients (p

The prognosis for hepatocellular carcinoma (HCC) remains dismal in terms of overall survival (OS), and its molecular pathogenesis has not been completely defined. Here, we report that expression of deubiquitylase ubiquitin-specific protease 7 (USP7) is higher in human HCC tissues than in matched peritumoral tissues. Ectopic USP7 expression promotes growth of HCC cells in vivo and in vitro. Mechanistically, USP7 overexpression fosters HCC cell growth by forming a complex with and stabilizing thyroid hormone receptor-interacting protein 12 (TRIP12), which induces constitutive p14ARF ubiquitination. Clinically, USP7 overexpression is significantly correlated with a malignant phenotype, including larger tumor size, multiple tumor, poor differentiation, elevated alpha-fetoprotein, and microvascular invasion. Moreover, overexpression of USP7 and/or TRIP12 correlates with shorter OS and higher cumulative recurrence rates of HCC. Conclusion: USP7 stabilizes TRIP12 by deubiquitination, thus constitutively inactivating p14ARF and promoting HCC progression. This represents a novel marker for predicting prognosis and a potential therapeutic target for HCC. (Hepatology 2015;61:1603-1614)

Intrahepatic cholangiocarcinoma (ICC) is an aggressive type of cancer, and its incidence and mortality rates are increasing worldwide. Mortalin is a highly conserved chaperone protein involved in multiple pathological and physiological processes, including anti-apoptosis, carcinogenesis and metastasis. The Bcl-2 family of proteins can be divided into pro-survival and pro-apoptotic members, including B-cell lymphoma 2 (Bcl-2) and Bcl-2-like protein 4 (Bax). The aim of the present study was to investigate the association between mortalin, Bcl-2 and Bax, as well as the prognostic significance of the combined expression of mortalin, Bcl-2 and Bax in ICC. Immunohistochemistry was used to determine the expression of mortalin, Bcl-2 and Bax in 116 ICC samples and to assess the association between expression of 3 markers and clinicopathological features of ICC patients. This revealed that ICC tumor tissues overexpressed mortalin and Bcl-2 and exhibited low expression of Bax in ICC tumor tissues compared with that in corresponding peritumoral samples. According to Pearson's correlation coefficient analysis, high expression of mortalin in ICC was positively correlated with Bcl-2 expression and negatively correlated with Bax expression. Furthermore, multiple linear regression analysis demonstrated that mortalin was positively associated with Bcl-2, but not with Bax, in patients with ICC. Patients with ICC exhibiting high expression of mortalin/Bcl-2 or low expression of Bax were closely associated with a malignant ICC phenotype, a relatively low overall survival rate and a high recurrence rate. Multivariate analysis indicated that mortalin and Bcl-2 were independent prognostic indicators for ICC patients. Meanwhile, the concomitant overexpression of mortalin and Bcl-2 and the low expression of Bax were independent markers for predicting a relatively poor prognosis of ICC. The overexpression of mortalin and Bcl-2 and/or the low expression of Bax are implicated in the anti-apoptotic effect and tumor progression of ICC. Mortalin or Bcl-2, or a combination of mortalin, Bcl-2 and Bax may be used to predict the prognosis of ICC, as well as potential therapeutic targets.

The 14-3-3 proteins are highly conserved molecules that are involved in many vital biologic processes and are associated with the progression of cancer. The role of 14-3-3ζ, a dimeric isoform of 14-3-3, in intrahepatic cholangiocarcinoma (ICC) was investigated in this study. The expression of 14-3-3ζ in tumour samples from patients with ICC was examined by Western blot and immunohistochemistry, and the correlation between its expression and various clinicopathological features was determined. Then, the capacity for invasion, migration and proliferation as well as the expression of epithelial-mesenchymal transition (EMT)-related markers in ICC cells were assessed after 14-3-3ζ depletion. Finally, the prognostic significance of 14-3-3ζ in patients with ICC was further evaluated by Kaplan-Meier and Cox regression analyses. The expression of 14-3-3ζ was significantly higher in ICC tissues compared to peritumoural tissues. High expression of 14-3-3ζ positively correlated with lymphatic metastasis and tumour-node-metastasis (TNM) stage. The inhibition of 14-3-3ζ expression was able to impair the invasion, migration and proliferation of ICC cells in vitro. The expression of 14-3-3ζ was significantly correlated with the expression of the EMT-related markers snail and E-cadherin in ICC samples. Moreover, the down-regulation of 14-3-3ζ also decreased the phosphorylation of extracellular signal-regulated kinase (ERK) in ICC cells. Clinically, patients with ICC with high 14-3-3ζ expression demonstrated a poor prognosis in terms of a short overall survival and a high recurrence rate of the disease. A multivariate analysis revealed that 14-3-3ζ overexpression was an independent prognostic indicator for patients with ICC. 14-3-3ζ may enhance the invasive and proliferative capacity of tumour cells and thus prompt the progression of ICC via the activation of ERK signalling and the induction of EMT. The overexpression of 14-3-3ζ may be used as a prognostic biomarker and therapeutic target in patients with ICC.

The aim of the present study was to investigate the expression levels of transforming growth factor-β (TGF-β) receptor type II (TβRII) and DPC4/Smad4 in the TGF-β signaling pathway and the importance of these expression levels in non-small cell lung cancer (NSCLC). The mRNA and protein expression levels of TβRII and DPC4/Smad4 were detected by reverse transcription-quantitative polymerase chain reaction and western blotting, respectively, in NSCLC and control nonlesional lung tissues of 60 patients. The protein expression levels of DPC4/Smad4 were detected by immunohistochemistry in paraffin-embedded samples of NSCLC. In addition, the correlations among the expression levels of TβRII and DPC4/Smad4 and their association with the clinical and pathological features of NSCLC were analyzed. The expression levels of TβRII and DPC4/Smad4 in NSCLC tissues were significantly lower when compared with the control nonlesional lung tissues (P

This paper presents development length comparison between Chinese and Australian codes study on HRB500 steel bars in concrete. After introducing the existing code provisions about the anchorage length and experimental investigation of tensile HRB500 steel bars, the calculation approaches in different codes are compared with the test data. Additionally, the development length of tensile HRB500 reinforcement bars in AS3600-2001, Proposed Revision of AS3600, GB50010-2010 and the calculation equation proposed from test results are compared with each other in beams and slabs. It is proved that AS3600-2001 is inconsistent with other standards and the tests results and needs to be revised, while the Proposed Revision of AS3600 provided a good agreement with the test data and brings the code into line with other international codes, particularly Eurocode 2. Moreover, the Proposed Revision of AS3600 is a significant improvement on the method in AS3600-2001. The anchorage length formula in GB50010-2010 can still be used for designing HRB500 steel bar.

In this paper, a generalized irreversible heat pump driven by steam turbine cycle model was established by taking account of the heat resistances, heat leak and irreversibility due to the internal dissipation of the working substance. The heat transfer between the heat reservoir and the working substance is assumed to obey the linear (Newtonian) heat transfer law, and the overall heat transfer surface area of the assumed to be constant. The fundamental optimal relations between the heat load, the thermal coefficient of the system and the temperature difference of the evaporator are obtained. Moreover, the effects of the cycle parameters on the characteristics of the cycle are studied by numerical example. The results obtained herein have realistic significance and may provide some theoretical guidance for the performance improvements and optimizations of heat pumps driven by steam turbine.

Considering the privacy issues on social network, a variety of anonymous techniques have been proposed, but these techniques neglect some differences among individuals in their demand for privacy protection. With the development of internet technology, the number of social network individuals increases yearly, and network data are poised for a massive change in trends. Motivated by this, we specify three levels of privacy information for victim individuals and propose a personalised k-degree-m-label (PKDML) anonymity model. Furthermore, we design and implement a distributed and personalised k-degree-m-lable (DPKDML) anonymisation algorithm, which takes advantage of the 'vertex-centric' GraphX programming model to complete the entire anonymous process by multiple message passing and node value updating. Finally, we conduct experiments on real social network datasets to evaluate the DPKDML, The experimental results show that our methods may overcome the shortcomings of traditional methods in processing massive data, and reduce anonymous costs and increase data utility.

In this paper, a moving object detection method based on image blocking and improved Particle Filter is proposed. Firstly, each frame in a video is partitioned into blocks and a low-dimensional descriptor is extracted for each block; Secondly, each block is sequentially passed through two classifiers, improved Particle Filter and temporal correlation check. Finally, foreground object is detected by integrating the classification results of all blocks. We use two-stage classifier to effectively minimize the number of false positive in the detected foreground object. Experiments show that the method proposed in this paper is feasible both in qualitative and quantitative analysis so it can be used in moving object detection effectively.

The electrochemical behavior of a new magnesium alloy (AZ61) containing rare earth elements-cerium (Mg-Al-Zn-Mn-Ce alloys) was investigated in 3% NaCl electrolyte using electrochemical methods such as linear sweep voltammetry, Tafel curves and electrochemical impedance spectroscopy. Scanning electron microscopy was used to characterize the surface morphologies of magnesium and its alloys. The results shows that compared with that of the most commonly used Mg alloy–AZ61, the cerium containing magnesium alloy exhibited higher electrochemical activity, and higher corrosion resistance. The electrochemical activity of Mg-Al-Zn-Mn-Ce was higher than that of Mg and Mg-Al-Zn-Mn-Ce alloys in 3% NaCl. The corrosion resistive order decreased in the following sequence: Mg-Al-Zn-Mn-Ce > Mg-Al-Zn-Mn > Mg. The electrolytes favored anodic magnesium oxidation, but the alloying element of Ce facilitated the formation of dense passive films on alloy surfaces.

We have modified the mitral repair technique in infants and small children by using autologous pericardial strips to treat mitral regurgitation resulting from a dilated mitral annulus. Our results demonstrate that this technique maintains stability and flexibility of the mitral annulus and decreases the risk of mitral stenosis.

Aims The prognosis of patients with intrahepatic cholangiocarcinoma (ICC) remains poor in terms of overall survival (OS) and recurrence rate. Mortalin, a stress chaperone, has been reported to be involved in carcinogenesis and metastasis. However, its role in ICC has not been defined. Methods Mortalin expression in tumour samples from patients with ICC was examined by Western blot and immunohistochemistry, and correlation between its expression and clinicopathological features was assessed. In addition, invasion, migration proliferation and apoptosis, and the expression of epithelial-mesenchymal transition (EMT)-related markers in ICC cells were assessed after mortalin depletion. Finally, the prognostic significance of mortalin in patients with ICC was further evaluated by Kaplan–Meier and Cox regression analysis. Results We provide evidence that expression of mortalin in human ICC tissues is higher than that in matched peritumoural tissues. The interference of mortalin expression inhibited the proliferation and invasion of ICC cells in vitro. Mechanistically, inhibition of mortalin expression in ICC cells upregulated E-cadherin expression and decreased vimentin and snail expression. Clinically, a high level of mortalin in ICC samples was associated with loss of E-cadherin, and increased expression of vimentin and snail. Patients with ICC and high mortalin expression had a shorter OS and a higher recurrence rate. Multivariate analysis revealed that mortalin overexpression was an independent prognostic indicator for patients with ICC. Conclusions Mortalin may promote cell proliferation and invasion via induction of EMT of ICC cells. A high level of mortalin may be used as a prognostic biomarker and therapeutic target for patients with ICC.

With the emergence of large-volume and high-speed streaming data, traditional techniques for mining closed frequent itemsets has become inefficient. Online mining of closed frequent itemsets over streaming data is one of the most important issues in mining data streams. In this paper, a combinative data structure is designed by using an effective bit-victor to represent items and an extended dictionary frequent item list to record the current closed frequent information in streams. For tremendous reduction of search space, some new search strategies are proposed to avoid a large number of intermediate itemsets generated. Meanwhile, some new pruning strategies are also proposed for the purpose of efficiently and dynamically maintaining of all the closure check operations. Experimental results show that the method proposed is efficient in time, with sound scalability as the number of transactions processed increases and adapts rapidly to the changes in data streams.

Software reliability assessment plays an important role in the software reliability engineering, but the traditional software assessment method mostly focuses on evaluation by use of failure data which is gained only after testing or usage in the late phase of the software life cycle. Thus people hope to get every stage’s information about the software’s reliability which is taken as the reference or accordance to guide the software’s design, analysis and testing and so on. The correlative factors of various products, processes and resources in the whole software life cycle have a very important effect on software reliability and, uncertainty exists in the software reliability evaluation to some extent. Fuzzy evaluation has a strong applicability for solving the fuzzy problem and an approach for software reliability qualitative evaluation based on fuzzy evaluation was proposed in this paper. Firstly the rationale and process of this method were described in detail, then a case study was implemented. The experimental results showed that the approach was feasible and effective.

As the requirements of traffic functions and urban landscape, a reinforced concrete arch- deck bridge is designed with archaized connective corridors. The main dimensions and drawings are introduced and the static analytical results of the bridge are discussed in this paper. The numerical model was built by the integrated solution system for bridge and civil engineering- MIDAS/Civil. The results show that the temperature of bridge and the horizontal shift at arch toes due to sedimentation play important roles sensitively influencing the bending moments and displacements of control sections. Compared numerical results of the bridge only with dead loads, the maximum bending moments and displacements would reach 3.81 times and 6.52 times respectively. The axial force of arch is mainly resulted from dead loads, which gradually increases from arch crown to arch toes. The design and numerical results would give some references to similar bridges.

This paper presents the test results of 2 prestressed concrete beams with manufactured sand under cyclic loading, the concrete is made of manufactured sand to replace natural sand with different replacement rates of 50% and 100%. The stress of re-bars and prestressed strands and the strain of manufactured sand concrete are discussed, the rigidity and cracking of the prestressed beams under cyclic loading are analyzed, and the flexural behavior of the beams after 2.5 million times cyclic loading are also discussed and analyzed. The results show that the fatigue behavior of the prestressed concrete beams with manufactured sand to replace the natural sand under cyclic loading is satisfied, the flexural bearing capacity of the prestressed beams after 2.5 million times cyclic loading is still sufficient. The results also show that the crack width and the mid-span deflection of the prestressed beams are in the allowable range at the serviceability limit state. This may be regarded as a reference for the applications of manufactured sand in the prestressed concrete structure .

Cold-rolled ribbed steel wires with improved elongation is a new type of cold-rolled steel wires, this paper presents the test results of 6 concrete slabs reinforced by the cold-rolled ribbed steel wires with improved elongation under flexure, the flexural characteristics, bearing capacity, deflection and cracking of these slabs under loading are analyzed, the formulas for calculating the flexural bearing capacity, the deflection and crack width of the slabs are suggested. This may be as a reference for the applications of this new type of cold-rolled steel wires, and for the revision of Chinese Code Technical specification for concrete structures with cold-rolled ribbed steel wiresJGJ95-2011.

Now, the resource of high quality natural sand is lacking. Manufactured sand is broken by rocks or tailings, which could be used in the mixture of cement concrete as fine aggregate instead of natural sand. The use of manufactured sand in concrete structure has become more and more important. It can make construction industry develop with environmentally friendliness and sustainability. In this paper, the elastic modulus strength was compared between manufactured sand concrete and ordinary concrete by testing on concrete specimens using the manufactured sand, the replacement ratios of manufactured sand by mass to the natural sand are 0, 0.30, 0.5, 0.70 and 1.0 respectively. The experimental research showed that the elastic modulus strength of manufactured sand concrete was stronger than that of natural sand and it had an increasing trend with the increasing of replacement rate and age but the elastic modulus of replacement rate of 50% was largest. This showed that the optimum replacement ratio was 50%.The fitting relations between elastic modulus and compressive strength was different with that of ordinary concrete. The relationship equation between elastic modulus strength and compressive strength of manufactured sand concrete was recommended in this paper. This could be used as a reference for the applications of the concrete using the manufactured sand as a fine aggregate in engineering.

This paper presents the test results of 6 reinforced concrete beams with recycled aggregates of construction waste under flexure, the beams were made of two strength grade concrete of and with different recycled aggregate replacement rate. The results indicate: the ultimate bearing capacity of recycled concrete beams with natural aggregate concrete beams are almost the same, and can meet the requirements of chinese code; The cracking resistance of the reinforced recycled concrete beams is slightly less than that of the beams with natural aggregates, the influence of recycled aggregate replacement rate to cracking resistance is not obvious. Recycled concrete beam crack load the calculated value is greater than the measured value, should carry out the theoretical value adjustment. Reinforced concrete beams is one of concrete structures, its the most common and most important component, Study of flexural property of reinforced concrete for recycled concrete structure component in the popularization and application to have the important significance [. This paper presents the test results of 6 reinforced concrete beams with recycled aggregates of construction waste under flexure, the beams were made of two strength grade concrete of C30 and C40 and with different recycled aggregate replacement rate of 0%, 50% and 70%. These results may be as a reference for the application of the concrete with recycled aggregates of construction waste in engineering [2,3,.

The influence of the aperture of transducer on the photoacoustic imaging was analyzed in this paper. We used different lengths of the same absorber for experiments, and these results demonstrated that the signal intensity would become stronger with the increasing of length of the absorber. But it will remain the same when its length over a certain value.

In this paper, based on circular scanning configuration of photoacoustic tomography (PAT), we have tested the effects of the stepping angle of scanning and the relationship between the scanning range and the orientation of the absorber on the quality of the reconstructed image. The experiment results have shown that the reconstructed image would be influenced more by artifacts as the increase of the stepping angle of scanning, and for the limited-view scanning, while the relationship of the scanning path and the orientation of the absorber is parallel, compared with vertical, we can use the data got from less detecting position to reconstruct image clearly.