Molecular detection of epithelial-mesenchymal transition markers in circulating tumor cells from pancreatic cancer patients:Potential role in clinical practice

AIM To evaluate the clinical properties of three subpopulations of circulating tumor cells(CTCs) undergoing epithelial-mesenchymal transition(EMT) in pancreatic ductal adenocarcinoma(PDAC) patients.METHODS We identified CTCs for expression of the epithelial cell marker cytokeratin or epithelial cell adhesion molecule(EpCAM)(E-CTC), the mesenchymal cell markers vimentin and twist(M-CTC), or both(E/M-CTC) using the CanPatrol system. Between July 2014 and July 2016, 107 patients with PDAC were enrolled for CTC evaluation. CTC enumeration and classification were correlated with patient clinicopathological features and outcomes.RESULTS CTCs were detected in 78.5% of PDAC patients. The number of total CTCs ranged from 0 to 26 across all 107 patients, with a median value of six. CTC status correlated with lymph node metastasis, TNM stage, distant metastasis, blood lymphocyte counts, and neutrophil-to-lymphocyte ratio(NLR). Kaplan-Meier survival analysis showed that patients with ≥ 6 total CTCs had significantly decreased overall survival and progression-free survival compared with patients with < 6 total CTCs. The presence of M-CTCs was positively correlated with TNM stage(P < 0.01) and distant metastasis(P < 0.01). Additionally, lymphocyte counts and NLR in patients without CTCs were significantly different from those in patients testing positive for each CTC subpopulation(P < 0.01).CONCLUSION Classifying CTCs by EMT markers helps to identify the more aggressive CTC subpopulations and provides useful evidence for determining a suitable clinical approach.

Linc00675 is a novel marker of short survival and recurrence in patients with pancreatic ductal adenocarcinoma

AIM: To detect linc00675 expression in pancreatic ductal adenocarcinoma(PDAC),to analyze the relationship between the expression level of linc00675 and the clinical pathological characteristics,to explore the biological functions of linc00675,and to determine whether linc00675 has independent prognostic value in PDAC.METHODS: We studied linc00675 expression among eight histologically confirmed PDAC tissue samples and four chronic pancreatitis tissue samples through microarray screening. RT-q PCR was conducted to further investigate linc00675 expression in PDAC cell lines as well as archived tissues from a large cohort of PDAC patients. The correlations between the level of lnc00675 and clinicopathological characteristics and survival in patients with pancreatic cancer were evaluated using Correlation analysis. Univariate andmultivariate analyses were conducted to predict whether lnc00675 expression is an independent prognostic and recurrence factor in patients with pancreatic cancer. After downregulating the expression of linc00675 through si RNA,MTT assay,flow cytometry,transwell assay and Western blot were used to explore the biological function of linc00675 in proliferation,invasion,and cell cycle progression of pancreatic cancer cells. The relative molecular expression levels of epithelial-mesenchymal transition were determined by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.RESULTS: The expression of Linc00675 in PDAC tissue samples was shown to be 672 times that in chronic pancreatitis tissue samples by microarray screening(P = 3.69 × 10-5). This finding was confirmed in tumor tissues from 90 patients with PDAC compared with adjacent normal tissue samples by quantitative RTPCR. We found that linc00675 overexpression positively correlated with lymph node metastasis(P = 0.005),perineural invasion(P = 0.006),and poor survival(P < 0.001). Univariate and multivariate analyses showed that linc00675 expression served as an independent predictor of overall survival(P = 0.009). Additionally,receiver operating characteristic curve analysis showed that high linc00675 might serve as a predictor of tumor progression within 6 mo to a year after surgery. In vitro functional analysis demonstrated that knockdown of linc00675 attenuated pancreatic cancer cell proliferation and invasion as well as induced S phase arrest. Suppression of linc00675 in pancreatic cancer cells resulted can reverse the progress of epithelialmesenchymal transition.CONCLUSION: Linc00675 may function as an oncogene during PDAC development,and its expression is an independent predictor of unfavorable prognosis in patients with PDAC.

Effect of Trimetazidine Dihydrochloride Tablets adjuvant therapy on inflammatory reaction, oxidative stress, vascular endothelial function and myocardial function in patients with coronary heart disease complicated with heart failure

Objective: To investigate the effects of Trimetazidine Dihydrochloride Tablets on inflammatory reaction, oxidative stress, vascular endothelial function and myocardial function in patients with coronary heart disease complicated with heart failure. Methods: A total of 98 patients with coronary heart disease and heart failure who met the criteria of the study were selected as the subjects, based on the random data table they were divided into the control group (n=49) and observation group (n=49), the patients in the control group were treated with Metoprolol Tartrate Sustained-release Tablets treatment, and the patients in the observation group were treated with Metoprolol Tartrate Sustained-release Tablets combined with Trimetazidine Dihydrochloride Tablets, the levels of inflammatory reaction, oxidative stress, vascular endothelial function and myocardial function indexes were compared between the two groups before and after treatment. Results: The difference of the CRP, TNF-α, MDA, SOD, NO, ET-1, LVEF, LVEDD and LVESD levels in the two groups before treatment were not statistically significant;Compared with the levels of the two groups before treatment, the two groups of CRP, TNF-α, MDA, ET-1, LVEDD and LVESD levels after treatment were significantly decreased, and the level of the observation group after treatment was significantly lower than those levels in the control group, the difference was statistically significant;The levels of SOD, NO and LVEF of the two groups after treatment were significantly higher than those in the same group before treatment, and the observation group levels [(88.09±7.51) U/ml, (72.58±14.64) mol/L, (48.34±5.09)% ] were significantly higher than the control group [(79.44±7.27) U/ml, (61.89±11.06) mol/L, (44.19±4.58)%], the difference was statistically significant. Conclusion: Trimetazidine Dihydrochloride Tablets in the treatment of coronary heart disease with heart failure can effectively inhibit the release of inflammatory factors, improve oxidative stress state, vascular endothelial function and myocardial function, has an important clinical value.

Development of a new 17-item Asthenopia Survey Questionnaire using Rasch analysis

AIM:To develop the 17-item Asthenopia Survey Questionnaire(ASQ)-17 by Rasch analysis,and to generate a predictiveness score.METHODS:Totally 739 participants were recruited and 680 were involved in the result analysis in this prospective,cross-sectional study.Three rounds of Rasch analysis were used to analyze the psychometric characteristics of items and options.RESULTS:Phase 1 assessed the original ASQ-19,adjusted the item scoring mode to a four-point Likert response rating scale and combined the 18th and 19th items into a new item.Phase 2 deleted the 11th item.Phases 3 and 4 assessed the new ASQ-17.All the evaluation indexes of ASQ-17 were acceptable.The Infit and Outfit MnSq values of items were 0.67-1.48,the variance explained by the principal component and the unexplained variance explained by the first contrast were 53.90%-59.40%and 1.50-1.80 in three dimensions.The curve peaks of scores in each dimension were separated and in the same order.The PSR and PSI values were 2.80 and 0.89,respectively.The mean scores of dimensions A(9.5±4.1 vs 3.5±3.2),B(7.3±3.3 vs 2.5±2.7),C(4.3±2.2 vs 1.4±2.0)and total(21.1±8.1 vs 7.4±7.0)in asthenopia participants were significantly higher than those without asthenopia(all P<0.001).The area under the curve in two groups was 0.899(P<0.001).Youden’s index was up to the maximum value of 0.784 when the cut-off value was 12.5.CONCLUSION:ASQ-17 has stronger option sorting and suitability than ASQ-19.It is an effective assessment tool for asthenopia with an optimal cut-off threshold value of 12.5,which is suitable for diagnosis and curative effect evaluation.

Preoperative evaluation with T-staging system for hilar cholangiocarcinoma

AIM： To investigate the clinical value of T-staging system in the preoperative assessment of hilar cholangiocarcinoma. METHODS： From March 1993 to January 2006, 85 patients who had cholangiocarcinoma diagnosed by operative tissue-biopsy were placed into one of three stages based on the new T-staging system, and it was evaluated the resectability and survival correlated with T-staging. RESULTS： The likelihood of resection and achieving tumor-free margin decreased progressively with increasing T stage （P 〈 0.05）. The cumulative 1-year survival rates of T1, T2 and T3 patients were 71.8%, 50.8% and 12.9% respectively, and the cumulative 3-year survival rate was 34.4%, 18.2% and 0% respectively; the survival of different stage patients differed markedly （P 〈 0.001）. Median survival in the hepatic resection group was greater than in the group that did not undergo hepatic resection （28 mo vs 18 mo; P 〈 0.05）. The overall accuracy for combined MRCP and color Doppler Ultrasonagraphy detecting disease was higher than that of combined using CT and color Doppler Ultrasonagraphy （91.4% vs 68%; P 〈 0.05 ）. And it was also higher in detecting port vein involvement （90% vs 54.5%; P 〈 0.05）.CONCLUSION： The proposed staging system for hilar cholangiocarcinoma can accurately predict resectability, the likelihood of metastatic disease, and survival. A concomitant partial hepatectomy would help to attain curative resection and the possibility of longterm survival. MRCP/MRA coupled with color Doppler UItrasonagraphy was necessary for preoperative evaluation of hilar cholangiocarcinoma.（OR = 2.46, 95% CI = 0.98-6.14）, and a significantly elevated risk of developing esophageal cancer among alcohol drinkers among alcohol drinkers （OR = 9.86, 95% CI = 3.10-31.38）. CONCLUSION： ADH2 and ALDH2 genotypes areassociated with esophageal cancer risk. ADH2＊1 allele and ALDH2＊2 allele carriers have a much higher risk of developing esophageal cancer, especially among alcohol drinkers.

Low RASSF6 expression in pancreatic ductal adenocarcinoma is associated with poor survival

AIM:To analyze RASSF6 expression in pancreatic ductal adenocarcinoma(PDAC) and to determine whether RASSF6 has an independent prognostic value in PDAC.METHODS:We studied RASSF6 expression in 96 histologically confirmed PDAC samples and 20 chronic pancreatitis specimens using immunohistochemistry and real-time quantitative reverse transcription-PCR.PDAC issues were then classified as RASSF6 strongly positive,weakly positive or negative.RASSF6 mR NA and protein expression in PDAC samples with strong positive staining was further evaluated using real-time PCR and Western blot analysis.Lastly,correlations between RASSF6 staining and patients' clinicopathological variables and outcomes were assessed.RESULTS:RASSF6 was negatively expressed in 51(53.1%) PDAC samples,weakly positively expressed in 29(30.2%) and strongly positively expressed in16(16.7%),while its expression was much higher in para-tumor tissues and chronic pancreatitis tissues.Positive relationships between RASSF6 expression and T-stage(P = 0.047) and perineural invasion(P = 0.026) were observed.The median survival time of strongly and weakly positive and negative RASSF6 staining groups was 33 mo,15 mo and 11 mo,respectively.Cox multivariate analysis indicated that RASSF6 was an independent prognostic indicator of overall survival in patients with PDAC.A survival curve analysis revealed that increased RASSF6 expression was correlated with better overall survival(P = 0.009).CONCLUSION:RASSF6 expression is an independent biomarker of an unfavorable prognosis in patients with PDAC.

Study of Test Flow Optimization Method in Radar Fault Isolation

In order to optimize test flow after the default flow is modified by a tester, a new software framework for the radar fault isolation is illustrated. This framework separates all mapping algorithms from test flows so as to modify flow and to insert mapping algorithm dynamically in testing process. Based on this framework, a kind of optimization method of test flow is proposed and studied. By defining an objective function, we can evaluate all candidate test flows so as to get an optimized flow. An example explains how to search the flow from candidate flows.

Combined effect of microplastic and triphenyltin:Insights from the gut-brain axis

Microplastics(MPs),an emerging group of pollutants,not only have direct toxic effects on aquatic organisms but also cause combined toxicity by absorbing other pollutants.Triphenyltin(TPT),one of the most widely used organotin compounds,has adverse effects on aquatic organisms.However,little is known about the combined toxicity of MPs and TPT to aquatic organisms.To investigate the individual and combined toxicity of MPs and TPT,we selected the common carp(Cyprinus carpio)for a 42-day exposure experiment.Based on the environmental concentrations in a heavily polluted area,the experimental concentrations of MPs and TPT were set at 0.5 mg L^(−1) and 1μg L^(−1),respectively.The effects of MPs combined with TPT on the carp gut-brain axis were evaluated by detecting gut physiology and biochemical parameters,gut microbial 16S rRNA,and brain transcriptome sequencing.Our results suggest that a single TPT caused lipid metabolism disorder and a single MP induced immunosuppression in carp.When MPs were combined with TPT,the involvement of TPT amplified the immunotoxic effect induced by MPs.In this study,we also explored the gut-brain axis relationship of carp immunosuppression,providing new insights for assessing the combined toxicity of MPs and TPT.At the same time,our study provides a theoretical basis for evaluating the coexistence risk of MPs and TPT in the aquatic environment.

Association between Polymorphism of Interleukin-23 Receptor and Hashimoto＇s Thyroiditis in Chinese Han Population of Shandong

Objective：The interleukin-23 receptor （IL-23R） has been shown to be associated with autoimmune diseases in many different populations.This study aimed to investigate the association between IL-23R gene polymorphism and susceptibility to Hashimoto＇s thyroiditis （HT） in Chinese Han population of Shandong.Methods：A case-control cohort study was performed in 145 HT patients from First People＇s Hospital of Jining between February 2010 to October 2013 and 150 healthy controls.Two single nucleotide polymorphisms located in the promoter region ofIL-23R gene （rs 17375018 and rs7517847） were examined by polymerase chain reaction-restriction fragment length polymorphism analysis.Hardy-Weinberg equilibrium was performed using the Chi-square test.Genotype frequencies were estimated by direct counting,and allele and genotype frequencies between patients and controls were analyzed by the Chi-square test.Results：The rs 17375018 GG genotype and the G allele were significantly increased in HT patients compared with healthy controls （P =0.034 and P =0.013,respectively）.No association was identified between HT patients and healthy controls in rs7517847.Conclusion：The study demonstrated that polymorphism of IL-23R gene rs17375018 is highly associated with HT in Chinese Han population of Shandong,suggesting that IL-23R gene polymorphism （rs 17375018 G） may play a critical role in susceptibility to HT.

Joint Design of Clustering and In-cluster Data Route for Heterogeneous Wireless Sensor Networks

A heterogeneous wireless sensor network comprises a number of inexpensive energy constrained wireless sensor nodes which collect data from the sensing environment and transmit them toward the improved cluster head in a coordinated way. Employing clustering techniques in such networks can achieve balanced energy consumption of member nodes and prolong the network lifetimes.In classical clustering techniques, clustering and in-cluster data routes are usually separated into independent operations. Although separate considerations of these two issues simplify the system design, it is often the non-optimal lifetime expectancy for wireless sensor networks. This paper proposes an integral framework that integrates these two correlated items in an interactive entirety. For that,we develop the clustering problems using nonlinear programming. Evolution process of clustering is provided in simulations. Results show that our joint-design proposal reaches the near optimal match between member nodes and cluster heads.

Wear-resistant coating spraying technique for wear-proofing cover of coke dry quenching furnace

During the operation of a coke dry quenching system of coking plant, wear-proofing cover of the boiler was seriously eroded, and even the ceiling tube of the boiler was worn and torn. Thus, attentions were paid to the research, development and application of the wear-resistant coating spraying technique for wear-proofing cover of the coke dry quenching furnace. The Cr3C2–NiCr coatings for wear-proofing cover were fabricated via supersonic arc spraying process, and the residual coating of wear-proofing cover would be sampled for being analyzed after two years. The coating presents a dense microstructure with few pores and micro-cracks (the porosity is 4.65%). After thermal spraying of the wear-proofing cover, the hardness and the wear resistance of the surface are improved. The remaining coating is continuous and compact, with an average thickness of about 147μm. Transverse micro-cracks parallel to the spreading direction of the coating surface are displayed on different areas of the coating, which demonstrates its serious erosion wear effect. The main chemical component of white zone is Fe–18Cr–Ni. The Cr level of light gray phase is about 77.57 wt.% and the O level is 22.43 wt.%. And the main chemical components of dark gray phase are C, O, Al, Si and Ca. X-ray diffraction patterns were adopted to implement phase analysis on the surface of coating sample, which indicated that the coating was composed of a large amount of Cr3O2 and a small amount of metal Cr.

Molecular insights of organochlorine biocide-induced toxicity in zebrafish:Whole-adult-organism toxicogenomics,targeted gene expression and histological analyses

Organochlorine biocides （OCBs）, such as lindane and dichlorodi- phenyltrichloroethane （DDT）, have been used for control of vector- borne diseases in public health and for insect pest control in agri- culture. These chemicals are extremely persistent in the environ- ment and they tend to bioaccumulate via food chains, thus posing risk to human and ecological health （Cooney et al., 2010; Wang et al., 2013）. Due to their persistence in the environment and continuous use in certain countries despite the banning of some of these chemicals, monitoring and studies of OCBs have continued intensely and remain relevant in foreseeable future （ Cohn et al.,