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Purpose To determine the ability of prostatic artery embolization (PAE) to achieve freedom from catheterization in patients with acute urinary retention (AUR) caused by benign prostatic hyperplasia (BPH). Materials and Methods This retrospective single-center study was performed between June 2014 and March 2019 in patients with lower urinary tract symptoms (LUTS) caused by BPH. PAE was performed in 154 eligible patients, of which 76 suffered from spontaneous AUR and had indwelling catheters placed and kept until the procedure, owing to clinical failure in the removal of the previous intermittent catheter. Each patient was followed for at least 12 months. The first trial without catheter was performed 3 days after PAE. Successful catheter removal within the first 30 days after PAE was considered a clinical success. The rate of patients free from catheterization, LUTS relief, prostate volume, and adverse events was recorded. Results Clinical success was achieved in 70 (92.1%) patients. The rates of freedom from catheterization were 90.3% (65/72), 83.3% (60/72), and 80.6% (58/72) at 3-, 6-, and 12-months follow-up, respectively. The median elapsed time from PAE to catheter removal was 10 days. However, 18 patients needed further interventions. Symptom scores revealed a continuous improvement in urinary symptoms. The mean prostate volume showed a statistically significant decrease at 3 and 12 months compared with its baseline value. No severe adverse events occurred. Conclusions PAE can achieve freedom from catheterization in patients with AUR caused by BPH.

To evaluate the safety and efficacy of prostatic artery embolization (PAE) in patients with recurrent lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia (BPH) who underwent a previous transurethral resection of the prostate (TURP).This retrospective study analyzed 15 of 19 patients who underwent PAE for recurrent LUTS after TURP between February 2014 and April 2019. The technical and clinical success rates and complications related to the procedure were recorded. International Prostate Symptom Score (IPSS), quality of life (QoL), and prostatic volume (PV) were evaluated at baseline and 3- and 12-mo follow-up.The intervals from TURP to recurrent symptoms and from TURP to PAE were 4.3 y ± 3.2 and 5.6 y ± 3.8, respectively. Technical success was achieved in all patients. The clinical success rate for LUTS relief at 12 mo was 93.3% (14 of 15). IPSS significantly reduced from 22.5 ± 4.1 at baseline to 9.9 ± 4.9 at 12-mo follow-up, and QoL score improved from 4.7 ± 1.0 to 2.1 ± 1.1 (P.05 for both). There was a significant mean reduction of 26.6% in PV at 12 mo, improving from 100.7 cmPAE may be a safe and effective treatment option for the management of recurrent LUTS secondary to BPH in patients who have previously undergone TURP.

To explore the efficacy of prostatic artery embolization (PAE) for lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia (BPH) during long-term follow-up and analyze predictors related to LUTS recurrence.This was a single-center retrospective study involving 125 BPH patients with LUTS who underwent PAE from February 2014 to February 2020. The median follow-up was 36 months. Clinical success was defined as reductions in the International Prostate Symptom Score (IPSS) and quality of life (QoL) score and no need for any other treatment for LUTS; otherwise, it was regarded as a clinical failure. Recurrence was defined as a clinical failure that occurred after an initial success. Cumulative clinical success rates, recurrence rates and re-intervention rates were evaluated. Friedman test was performed to compare differences in IPSS, QoL and prostatic volume (PV) among baseline and follow-up times. Predictors for LUTS recurrence were analyzed with the univariate and multivariate Cox regression model.Technical success (bilateral PAE) rate was 92.8% (116/125). Significant differences in IPSS, QoL and PV were observed between baseline and follow-up time points (P 0.001). The cumulative clinical success rates at 2, 3, 4 and 5 years were 82.4%, 65.5%, 52.4% and 37.4%. The cumulative recurrence rates and re-intervention rates at 1, 2 and 5 years were 6.8%, 12.7%, 60.4% and 5.9%, 10.2%, 50.8%, respectively. Unilateral PAE was an significant predictor of recurrence (P 0.05).PAE is an effective treatment option for LUTS. Unilateral PAE is a significant independent predictor of LUTS recurrence.

Accurately predicting the spatial distribution of soil organic matter (SOM) content is of great significance for improving soil quality and improving the level of regional soil management. In order to explore the optimal model for predicting the SOM content of farmland in the Weibei Dryland of Shaanxi Province, the influence factors closely related to SOM content were selected as the modeling covariables, and a geographic detector, the ordinary kriging method (OK), geographic weighted regression model (GWR), partial least squares regression model (PLS), geographically weighted regression extended model (GWR

Objective To explore whether suberoylanilide hydroxamic acid (SAHA, br. Vorinostat, a kind of histone deacetylase inhibitor) can improve the diastolic function of restrictive cardiomyopathy (RCM) mice by up-regulating the expression of wild type cardiac troponin I (WT-cTnI). Methods Sixteen male cTnI R193H mice were employed and divided into SAHA group (n=6), dimethyl sulfoxide (DMSO) group (n=5) and control groups (n=5). Mice were subcutaneously injected with 50mg/kg SAHA in SAHA group, with 1ml/kg DMSO in DMSO group and with 1ml/kg normal saline in control group, respectively. The total treatment duration lasted for 56 days. Western blotting was performed to determine the levels of acetylated histone H3 (acH3) and the expression of cTnI. The transcription levels of Tnni3 were detected with RT-qPCR. The levels of acH3 in Tnni3 promoter key area were detected with chromatin immunoprecipitation (ChIP). The data of diastolic function measurements were collected using high frequency echocardiography. Results Compared to control group, the acH3 levels in nucleus and in Tnni3 promoter key area increased significantly in SAHA group (P0.05), but the specific acH3 level in Tnni3 promoter key area increased significantly in SAHA group compared to that in DMSO group (P

Cardiac diastolic dysfunction (CDD) is the most common form of cardiovascular disorders, especially in elderly people. Cardiac troponin I (cTnI) plays a critical role in the regulation of cardiac function, especially diastolic function. Our previous studies showed that cTnI‐low expression induced by histone acetylation modification might be one of the causes that result in diastolic dysfunction in ageing hearts. This study was designed to investigate whether epigallocatechin‐3‐gallate (EGCG) would modify histone acetylation events to regulate cTnI expression and then improve cardiac functions in ageing mice. Our study shows that EGCG improved cardiac diastolic function of aged mice after 8‐week treatment. Low expression of cTnI in the ageing hearts was reversed through EGCG treatment. EGCG inhibited the expression of histone deacetylase 1 (HDAC1) and HDAC3, and the binding levels of HDAC1 in the proximal promoter of cTnI. Acetylated lysine 9 on histone H3 (AcH3K9) levels of cTnI's promoter were increased through EGCG treatment. Additionally, EGCG resulted in an ascent of the binding levels of transcription factors GATA4 and Mef2c with cTnI's promoter. Together, our data indicate that EGCG may improve cardiac diastolic function of ageing mice through up‐regulating cTnI by histone acetylation modification. These findings provide new insights into histone acetylation mechanisms of EGCG treatment that may contribute to the prevention of CDD in ageing populations.

To detect the development of monocytes and proliferative macrophages in atherosclerosis of ApoE

The 2014–2018 angiograms of 58 patients with prostate cancer were retrospectively analyzed to illustrate angiographic findings during prostatic artery embolization. Arteriovenous fistulae were observed in 6 patients (6/58, 10.3%), with no difference between patients with or without prior iodine-125 seeds implantation (5/48, 10.4% vs 1/10, 10.0%; P > .05); tumor staining was not detected. The origins of the prostatic arteries included the internal pudendal artery (n = 45, 32.4%), the superior vesical artery (n = 38, 27.3%), the obturator artery (n = 28, 20.1%), the gluteal-pudendal trunk (n = 21, 15.1%), the inferior gluteal artery (n = 3, 2.2%), the accessory pudendal artery (n = 3, 2.2%), and the superior gluteal artery (n = 1, 0.7%).

Glioma is one of the most malignant forms of brain tumor, and has been of persistent concern due to its high recurrence and mortality rates, and limited therapeutic options. As a cardiac glycoside, ouabain has widespread applications in congestive heart diseases due to its positive cardiac inotropic effect by inhibiting Na+/K+-ATPase. Previous studies have demonstrated that ouabain has antitumor activity in several types of human tumor, including glioma. However, the exact underlying mechanism remains to be elucidated. The purpose of present study was to elucidate the effect of ouabain on human glioma cell apoptosis and investigate the exact mechanism. U-87MG cells were treated with various concentrations of ouabain for 24 h, following which cell viability and survival rate were assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The dynamic changes and cell motility were observed using digital holographic microscopy. Additionally, western blot analysis and high-content screening assays were used to detect the protein expression levels of phosphorylated (p-)Akt, mammalian target of rapamycin (mTOR), p-mTOR and hypoxia-inducible factor (HIF)-1α, respectively. Compared with the control group, ouabain suppressed U-87MG cell survival, and attenuated cell motility in a dose-dependent manner (P

With the continuous increase in the size and complexity of a real-time computer system, the use of formal verification methods in software development is also on the rise. The traditional formal verification method is not fully applicable to the development of actual system life cycle. Therefore, this paper presents a new real-time system verification method, It takes the deadlock timed Buchi automata as the medium, and translates the timed temporal logic into timed communicating sequential process language. The tock event is also joined, which can be directly used for the detection of refinement tool FDR. The method verifies the situation of deadlock. To establish the link between the conventional model checking and refinement model checking can well combine the advantages of both and improves system security and reliability.

To prepare the asiaticoside nanoemulsions (ASI-NEs) and asiaticoside nanoemulsions-based gels (ASI-NBGs), compare them with the commercial cream of asiaticoside (ASI-C) in terms of transdermal characteristics, and investigate the transdermal mechanism of ASI-NEs and ASI-NBGs. Their transdermal characteristics were studied by using Franz diffusion cells. The effect of topical ASI-NEs and ASI-NBGs on ultrastructure of rabbit skin was evaluated by using HE staining method. The localization and the permeation pathway of asiaticoside were visually investigated by using laser scanning confocal microscope (CLSM). The transdermal studies in vitro showed that the cumulative amount of ASI permeated from ASI-NEs and ASI-NBGs at 12 h after application were (3 504.30±180.93), (1 187.40±128.88) μg·cm⁻² respectively, 6.57, 2.23 times of that in the control group of ASI-C; the drug deposition of ASI-NEs and ASI-NBGs in skin was (159.48±7.47), (120.53±5.71) μg·cm⁻² respectively, 5.93, 4.48 times of that of ASI-C. HE staining of the rabbit skin after application of ASI-NEs and ASI-NBGs showed that the epidermis structure was basically intact; stratum corneum was loosed and the keratin fragment was increased; at the same time, the gap of prickle cell was increased and the basal cells were arranged loosely. The study of CLSM showed that significant percutaneous enhancer effect was observed for ASI-NEs after the topical application of 6 h, as the fluorescent compound was penetrated in the dermis and diffused uniformly. The fluorescence area and the integral optical density (IOD) were 28.81, 32.51 times of that in the FITC aqueous solution group, respectively. The fluorescent preparations showed strong fluorescence in the epidermis, but weak in deeper layers; with the increase of treatment time, the fluorescence in deeper layer was increased and stronger in skin appendages. The prepared ASI-NEs and ASI-NBGs have good transdermal characteristics and the transdermal mechanism is related to breaking the ultrastructure of stratum corneum and penetrating by the path of skin adnexa.

Zing finger protein 580 (ZFP580) is a novel Cys2-His2 zinc-finger transcription factor that has an anti-apoptotic role in myocardial cells. It is involved in the endothelial transforming growth factor‑β1 (TGF‑β1) signal transduction pathway as a mothers against decapentaplegic homolog (Smad)2 binding partner. The aim of the present study was to determine the involvement of ZFP580 in TGF‑β1‑mediated cytoprotection against chemical hypoxia‑induced apoptosis, using H9c2 cardiac myocytes. Hypoxia was chemically induced in H9c2 myocardial cells by exposure to cobalt chloride (CoCl2). In response to hypoxia, cell viability was decreased, whereas the expression levels of hypoxia inducible factor-1α and ZFP580 were increased. Pretreatment with TGF‑β1 attenuated CoCl2‑induced cell apoptosis and upregulated ZFP580 protein expression; however, these effects could be suppressed by SB431542, an inhibitor of TGF‑β type I receptor and Smad2/3 phosphorylation. Furthermore, suppression of ZFP580 expression by RNA interference reduced the anti‑apoptotic effects of TGF‑β1 and thus increased CoCl2‑induced apoptosis. B‑cell lymphoma (Bcl)‑2‑associated X protein/Bcl‑2 ratio, reactive oxygen species generation and caspase‑3 activation were also increased following ZFP580 inactivation. In conclusion, these results indicate that ZFP580 is a component of the TGF-β1/Smad signaling pathway, and is involved in the protective effects of TGF‑β1 against chemical hypoxia‑induced cell apoptosis, through inhibition of the mitochondrial apoptotic pathway.

Train control system is a safety-critical system, which requires a thorough safety testing before they are deployed. But the traditional technology of test automation is not sound for safety test of safety-critical software system because of its just aiming at the system functional requirements. Focusing on the safety requirements, this paper addresses a new method to perform safety test of safety-critical system based on safety requirement by using model-based test. The method presents safety automata to express safety properties in term of safety requirements and extract safety model from the safety requirements. It also provides a new technique for automatic generation of safety test case suite. By using the method, we have designed an automatic testing framework and developed an automatic testing platform, which has been applied to standard testing the product of train control system in term of the requirements of Ministry of railways in china, and implement effectively the functional and safety test of the product of train control system in china.

Recent research has shown that the Na(+)/K(+)-ATPase alpha1 subunit is a novel anti-cancer target, which plays pivotal roles in malignant cell ion transport, metabolism, migration and signal transduction. The purpose of the present study was to investigate the anti-cancer effects of ouabain and Na(+)/K(+)-ATPase alpha1 small interfering ribonucleic acid (siRNA) on HepG2 cell proliferation, apoptosis and cell cycle, and to explore the molecular mechanisms. The expression of Na(+)/K(+)-ATPase alpha1 subunit in human hepatocellular carcinoma (HCC), normal liver tissues and human HCC line (HepG2, SMMC-7721 and Bel-7402) has been investigated. Using the ouabain and Na(+)/K(+)-ATPase alpha1 subunit siRNA, which target the Na(+)/K(+)-ATPase, we have evaluated the effects of inhibiting Na(+)/K(+)-ATPase alpha1 in human HepG2 cells with respect to cell proliferation, morphology, cell cycle, impact on intracellular Ca2++, reactive oxygen species (ROS) concentration, and correlated gene expression level on messenger ribonucleic acid (mRNA) and protein. Our data showed that the expression Na(+)/K(+)-ATPase alpha1 subunit in HCC tissues is higher than that in normal liver tissues. Ouabain and Na(+)/K(+)-ATPase alpha1 siRNA could inhibit HepG2 cell proliferation. Ouabain could induce HepG2 cell apoptosis and generate S phase arrest, and siRNA could enhance the anti-cancer effect of ouabain that induced HepG2 cells apoptosis via an intracellular Ca(2+) and ROS increase-mediated, and generated cell cycle S phase arresting by decreasing the CyclinA1/cyclin-dependent kinase 2 (CDK2)/proliferating cell nuclear antigen (PCNA) complex product and increasing the expression of cyclin-dependent kinase inhibitor 1A (P21(CIP1)). We believe that targeting of the Na(+)/K(+)-ATPase alpha1 subunit in human HCC cells could provide new sight into the treatment of HCC.

Many studies have shown the Na(+)/K(+)-ATPase (NKA) might be a potential target for anticancer therapy. Cardiac glycosides (CGs), as a family of naturally compounds, inhibited the NKA activity. The present study investigates the antitumor effect of ouabain and elucidates the pharmacological mechanisms of CG activity in liver cancer HepG2 cell using SILAC coupled to LC-MS/MS method. Bioinformatics analysis of 330 proteins that were changed in cells under treatment with 0.5 μmol/L ouabain showed that the biological processes are associated with an acute inflammatory response, cell cycle, oxidation reduction, chromosome segregation, and DNA metabolism. We confirmed that ouabain induced chromosome segregation disorder and S-cell cycle block by decreasing the expression of AURKA, SMC2, Cyclin D, and p-CDK1 as well as increasing the expression of p53. We found that the overexpression or inhibition of AURKA significantly reduced or enhanced the ouabain-mediated the anticancer effects. Our findings suggest that AURKA is involved in the anticancer mechanisms of ouabain in HepG2 cells.

Debate have two features. One is competitive, another is critical thinking. Competitiveness of debate can urge students to review and learn from past debate experience. And the necessary of refuting in debate will urge students to think critically on the opponent's opinion. Both of the two features can help students to learn how to react on some condition, which is the ability of response. Also by making statement and refuting them can also learn how to organize their words, which is the ability of externalization. In this paper, we propose a debate assistant system as a web application. By making use of the features of debate, the system can let students benefit from the online practice.

Pancreatic cancer is one of the leading causes of cancer-related death in Western countries. Bcl-x(L) is an anti-apoptotic factor of the Bcl-2 family, which is overexpressed in pancreatic cancer and its presence correlates with shorter patient survival. In this study, sequence-specific antisense oligonucleotides targeting the coding region of Bcl-x(L) were designed to examine whether apoptosis could be induced and chemosensitivity could be increased in pancreatic cancer cells. Five pancreatic cancer cell lines, Panc-1, MIA-PaCa-2, Capan-1, ASPC-1 and T3M4, were treated with Bcl-x(L) sense or antisense oligonucleotides and gemcitabine and the cell viability was examined by the SRB method. Apoptosis was determined using DAPI staining. In all examined pancreatic cancer cells, Bcl-x(L) expression was reduced after transfection of the antisense oligonucleotides. Cell death analysis using DAPI staining revealed that antisense, but not sense oligonucleotides caused apoptotic cell death. Furthermore, Bcl-x(L) antisense oligonucleotides enhanced the cytotoxic effects of gemcitabine in pancreatic cancer cells. Our results indicate that Bcl-x(L) antisense oligonucleotides effectively inhibited pancreatic cancer cell growth and caused apoptosis by reducing Bcl-x(L) protein levels. Bcl-x(L) antisense oligonucleotides also increased the chemosensitivity of pancreatic cancer cells, suggesting that Bcl-x(L) antisense therapy might be a potential future approach in this disease.

The recursive algorithms for depth-first traversal of a binary tree are widely expatiated upon in data structure textbooks. There are three depth-first traversal sequences for a binary tree, preorder, inorder, and postorder traversal sequences. My literature survey indicates the most references present the depth-first traversals algorithms as recursive ones. Some literatures have introduced the non-recursive algorithms only for one of these three traversal sequences of a binary tree. In this paper a general non-recursive algorithm for the depth-first traversal of a binary tree is proposed. This general non-recursive algorithm can visit every nodes of a binary tree in preorder, inorder, or postorder traversal sequence. The algorithm analysis shows that this new algorithm is efficient and easy to understand. The implementation of this new algorithm was done in C and the complete algorithm was tested.

To study the effect of pseudolaric acid B (PLAB) on cell proliferation and cycle of human prostate carcinoma DU-145 cells. method: Its inhibitory effect on the cell growth was measured by MTT method. Characteristics of cell death were determined by Hoechest 33342 staining. The cell cycle was detected by flow cytometry. The expressions of cyclin B1, cyclin D1 and CDK1 were detected by Real time-PCR and Western blot, respectively.PLAB notably inhibited DU-145 cell growth in a dose- and time dependent manner (P0.05). Its IC50 values of PLAB for DU-145 cells for 24, 48 and 72 h were 4.53, 2.39 and 2.08 micromol x L(-1), respectively. Having been treated with 5 micromol x L(-1) PLAB for 24 h, the cells showed such apoptosis characteristics as nuclei chromatin condensation and apoptotic body. With the increase in PLAB concentration, the proportion of G2/M phase cells strikingly increased in a dose- and time dependent manner (P0.05), meanwhile cyclin B1 and CDK1 showed over-expressions (P0.05), and the cyclin D1 showed under-expression (P0.05).PLAB can inhibit the growth of DU-145 cells and induce the cell cycle G2/M arrest, accompanied with the over-expression of cyclin B1 and CDK1, which may be related with its regulation cycle-associated protein degradation.

Supplemental Digital Content is available in the text, In experimental myocardial infarction (MI), a rise in cell counts of circulating monocyte subsets contributes to impaired myocardial healing and to atherosclerotic plaque destabilization. In humans, the prognostic role of monocyte subsets in patients suffering ST-elevation MI (STEMI) is still unclear. In the present study, we aimed to determine the kinetics of the 3 monocyte subsets (classical CD14++CD16–, intermediate CD14++CD16+, and nonclassical CD14+CD16++ monocytes), as well as the subset-specific monocyte–platelet aggregates (MPA), in acute STEMI followed by primary percutaneous coronary intervention (PCI), and their relationships with cardiovascular outcomes during a 2-year follow-up. Monocyte subsets and MPA were measured in 100 STEMI patients receiving primary PCI on days 1, 2, 3, 5, and 7 of symptom onset, which were compared with 60 stable coronary heart disease patients and 35 healthy volunteers. From day 1 to day 7, significant increases in the counts of CD14++CD16+ monocytes and CD14++CD16+ MPA were observed, with peak levels on day 2. During a median follow-up of 2.0 years, 28 first cardiovascular events (defined as cardiovascular death, nonfatal ischemic stroke, recurrent MI, need for emergency or repeat revascularization, and rehospitalization for heart failure) were recorded. After adjustment for confounders, CD14++CD16+ monocytosis (day 1 [HR: 3.428; 95% CI: 1.597–7.358; P = 0.002], day 2 [HR: 4.835; 95% CI: 1.106–21.13; P = 0.04], day 3 [HR: 2.734; 95% CI: 1.138–6.564; P = 0.02], and day 7 [HR: 2.647; 95% CI: 1.196–5.861; P = 0.02]), as well as increased levels of CD14++CD16+ MPA measured on all time points (days 1, 2, 3, 5, and 7), had predictive values for adverse cardiovascular events. In conclusion, our data show the expansion of the CD14++CD16+ monocyte subset during acute phase of STEMI has predictive values for 2-year adverse cardiovascular outcomes in patients treated with primary PCI. Future studies will be warranted to elucidate whether CD14++CD16+ monocytes may become a target cell population for new therapeutic strategies after STEMI.

Conventional methods for safety analysis have problems in terms of correctness and efficiency. Using formal methods and fault injection techniques can improve consistency and correctness for safety process throughout the system's safety lifecycle. Yet existed methods for fault injection and safety analysis mainly think about single faults injection for system, seldom do with fault injection for multiple faults. In the paper, we propose a novel fault injection algorithm for safety analysis, it can inject single faults and multiple faults to system model, automatically form extended system models, by safety verification for these models, verification results for fault injection are obtained, besides, we analyze the results of multiple faults, we can get the minimal combination of multiple faults which violate the safety. Compared with existed methods for safety analysis and fault injection, our new method provides an excellent way to form the minimal cut sets of fault trees, it has improved the completeness of fault injection, and reduced the time for safety verification and analysis.

Model-based test is the very useful method to verify software safety. But the traditional technology of model-based test is not suitable for safety test of safety-critical software system. This paper addresses a new approach to perform safety test of safety-critical system based on safety automaton model, which can express safety properties. By using the method, we have developed an automatic testing platform for train control center system safety test.

Safety is an important character of safety-critical software system. In order to validate the safety of safety-critical software system, people will use the method of testing. The key factor of the method is to model the system safety requirements and interaction constraints among components. This paper addresses a new approach to model the safety requirements by safety behavior model and perform safety test of safety-critical system based on it. By using the method, we have developed an automatic testing platform for train control system safety test.

Recent studies revealed the potential of Na(+)/K(+)-ATPase as a target for anticancer therapy and showed additional modes of action of cardiotonic steroids (CSs), a diverse family of naturally derived compounds, as inhibitors of Na(+)/K(+)-ATPase. The results from epidemiological studies showed significantly lower mortality rates in cancer patients receiving CSs, which sparked interest in the anticancer properties of these drugs. The present study was designed to investigate the anticancer effect of CSs (ouabain or cinobufagin) and to elucidate the molecular mechanisms of CS activity in hepatoma cell lines (HepG2 and SMMC-7721). Ouabain and cinobufagin significantly inhibited cell proliferation by attenuating the phosphorylation of extracellular regulated kinase (ERK) and down-regulating the expression of C-myc. These CSs also induced cell apoptosis by increasing the concentration of intracellular free calcium ([Ca(2+)](i)) and induced S phase cell cycle arrest by down-regulating the expression of Cyclin A, cyclin dependent kinase 2 (CDK2) and proliferating cell nuclear antigen (PCNA) as well as up-regulating the expression of cyclin dependent kinase inhibitor 1A (p21(CIP1)). Overexpression of ERK reversed the antiproliferation effect of ouabain or cinobufagin in HepG2 and SMMC-7721 cells. Currently, the first generation of CS-based anticancer drugs (UNBS1450 and Anvirzel) are in Phase I clinical trials. These data clearly support their potential use as cancer therapies.

Recent research has shown that the Na(+)/K(+)-ATPase alpha1 subunit is a novel anti-cancer target, which plays pivotal roles in malignant cell ion transport, metabolism, migration and signal transduction. The purpose of the present study was to investigate the anti-cancer effects of ouabain and Na(+)/K(+)-ATPase alpha1 small interfering ribonucleic acid (siRNA) on HepG2 cell proliferation, apoptosis and cell cycle, and to explore the molecular mechanisms. The expression of Na(+)/K(+)-ATPase alpha1 subunit in human hepatocellular carcinoma (HCC), normal liver tissues and human HCC line (HepG2, SMMC-7721 and Bel-7402) has been investigated. Using the ouabain and Na(+)/K(+)-ATPase alpha1 subunit siRNA, which target the Na(+)/K(+)-ATPase, we have evaluated the effects of inhibiting Na(+)/K(+)-ATPase alpha1 in human HepG2 cells with respect to cell proliferation, morphology, cell cycle, impact on intracellular Ca2++, reactive oxygen species (ROS) concentration, and correlated gene expression level on messenger ribonucleic acid (mRNA) and protein. Our data showed that the expression Na(+)/K(+)-ATPase alpha1 subunit in HCC tissues is higher than that in normal liver tissues. Ouabain and Na(+)/K(+)-ATPase alpha1 siRNA could inhibit HepG2 cell proliferation. Ouabain could induce HepG2 cell apoptosis and generate S phase arrest, and siRNA could enhance the anti-cancer effect of ouabain that induced HepG2 cells apoptosis via an intracellular Ca(2+) and ROS increase-mediated, and generated cell cycle S phase arresting by decreasing the CyclinA1/cyclin-dependent kinase 2 (CDK2)/proliferating cell nuclear antigen (PCNA) complex product and increasing the expression of cyclin-dependent kinase inhibitor 1A (P21(CIP1)). We believe that targeting of the Na(+)/K(+)-ATPase alpha1 subunit in human HCC cells could provide new sight into the treatment of HCC.

To investigate the in vitro anti-cancer effects of ouabain combined Na(+)/K(+)-ATPase alpha1 siRNA upon HepG2 cells and its molecular mechanism.The Na(+)/K(+)-ATPase alpha1 subunit expressions of human hepatoma tissue, normal liver tissue and human hepatoma cell lines (HepG2, SMC7721 and Bel7402) were determined. The cells received different treatments (0.03 micromol/L siRNA, 0.1 micromol/L ouabain and combination). The proliferation of HepG2 was observed by CCK-8 assay. The activity of Na(+)/K(+)-ATPase was measured. The HepG2 cell cycle distribution was detected by flow cytometry. The mRNA and protein levels of Na(+)/K(+)-ATPase alpha1 subunit, MAPK1, Cyclin A, CDK2, PCNA and P21WAF1 were detected by quantitative RT-PCR and Western blot.The gray value Na(+)/K(+)-ATPase alpha1 subunit in hepatoma tissue was 174.74 +/- 16.77 and 65.31 +/- 7.88 respectively in normal liver tissue. The Na(+)/K(+)-ATPase alpha1 subunit expression of hepatoma tissue was significantly higher than normal liver tissue(P0.05).The Na(+)/K(+)-ATPase alpha1 subunit expression level of HepG2 was higher than that in SMMC-7721 and Bel-7402 cells. The CCK-8 experiments demonstrated that siRNA, ouabain and combination could inhibit the HepG2 proliferation, and the combination group was different from the ouabain or siRNA group (P0.05). The 24, 48 and 72 h inhibitory rates of 0.03 micromol/L siRNA, 0.1 micromol/L ouabain and combination group were (17.4%, 20.3%, 24.3%), (37.5%, 44.3%, 51.2%) and (52.3%, 70.2%, 88.2%) respectively. The activity of Na(+)/K(+)-ATPase decreased in siRNA, ouabain and combination group. The S phase proportion of ouabain and combination group increased from 24.2% to 66.5% and 75.2% respectively. The 0.1 micromol/L ouabain and combination group could down-regulate the expression of Na(+)/K(+)-ATPsae alpha1, MAPK1, Cyclin A, CDK2, PCNA and up-regulate the expression of P21WAF1 in HepG2 cell.The Na(+)/K(+)-ATPase alpha1 siRNA combined ouabain inhibits the proliferation of HepG2 cells by decreasing the expression of MAPK1 and induces the cell cycle S arrest by decreasing the production of Cyclin A/CDK2/PCNA complex and increasing the expression of P21(WAF1).

To find one kind of peptide that will conjugate with ouabain and inhibit its biological function, and provide a new treatment strategy for primary hypertension.In this study, ouabain was used as a target to screen ouabain conjugated peptide (OCP) from Ph.D.-7 phage display peptide library. After 3 rounds of bio-panning, the products were identified by ELISA and DNA electrophoresis analysis and sequencing. Peptide was synthesized and analyzed the activity by radioligand binding assay. The inhibitory ratio of cell proliferation was measured by MTT and the cell morphology changing was measured by Hoechst 33342/PI staining. The expression of Na(+)-K(+)-ATPase alpha1-subunit and beta1-subunit were detected by RT-PCR and immunocytochemistry. The levels of the free intracellular Na(+) in EAhy926 cells were measured by laser confocal microscope.The ouabain conjugated peptide was found out, and it was occupied in 0.64 (9/14). The analysis of protein showed that the obtained peptides had no homology with Na(+)-K(+)-ATPase. The amino acid sequence of synthesized peptide was Arg-Cys-Met-Thr-Ser-Arg-Ser. There was binding activity between OCP and (3)H-ouabain. The MTT assay showed that OCP could reverse the inhibition action of ouabain on vascular endothelial EAhy926 cells in a dose and time-dependent manner. The number of apoptotic cells had significantly decreased detected by Hoechst 33342/PI staining. The results of RT-PCR and immunocytochemistry showed that OCP could inhibit the up-regulated expression of Na(+)-K(+)-ATPase alpha1-subunit and down-regulated expression of Na(+)-K(+)-ATPase beta1-subunit induced by ouabain in EAhy926 cells.The OCP could reverse the growth inhibition and death induction of ouabain in EAhy926 cells, which would provide the basis for studying the interaction between ouabain and Na(+)-K(+)-ATPase and explore novel anti-ouabain agents.

It is very important in HIS (Hospital Information System) to maintain the consistency and integrity of information. A transaction is a group of actions that must all be carried out successfully on one or more tables in a database before they are committed (made permanent). If any of the actions in the group fails, then all actions are rolled back (undone). In this paper, how to use transactions in Delphi in latent and apparent way is discussed in detail. Furthermore, a demonstration is given to show a method of using transactions in Delphi to guarantee the prescriptions being stored to the database or transmitted to another departments in intactness. This method could also be used to maintain database consistency and integrity when there is need for modifications to the data stored in the database.

Safety is a very important property for safetycritical software systems. We introduce an approach to test safety-critical software systems based on safety requirement by using model-based testing. The model is extracted from the safety requirement and system under test. In our approach, we use model checking techniques to generate test cases. To find appropriate test cases in less time, we divide the states spaces of model into different subsets. By using the approach, we have designed an automatic testing framework and developed an automatic testing platform, which has been used for testing the safety of high speed train control system in china

AIM: To screen and investigate the effective gRNAs against hepatitis B virus (HBV) of genotypes A-D. METHODS: A total of 15 gRNAs against HBV of genotypes A-D were designed. Eleven combinations of two above gRNAs (dual-gRNAs) covering the regulatory region of HBV were chosen. The efficiency of each gRNA and 11 dual-gRNAs on the suppression of HBV (genotypes A-D) replication was examined by the measurement of HBV surface antigen (HBsAg) or e antigen (HBeAg) in the culture supernatant. The destruction of HBV-expressing vector was examined in HuH7 cells co-transfected with dual-gRNAs and HBV-expressing vector using polymerase chain reaction (PCR) and sequencing method, and the destruction of cccDNA was examined in HepAD38 cells using KCl precipitation, plasmid-safe ATP-dependent DNase (PSAD) digestion, rolling circle amplification and quantitative PCR combined method. The cytotoxicity of these gRNAs was assessed by a mitochondrial tetrazolium assay. RESULTS: All of gRNAs could significantly reduce HBsAg or HBeAg production in the culture supernatant, which was dependent on the region in which gRNA against. All of dual gRNAs could efficiently suppress HBsAg and/or HBeAg production for HBV of genotypes A-D, and the efficacy of dual gRNAs in suppressing HBsAg and/or HBeAg production was significantly increased when compared to the single gRNA used alone. Furthermore, by PCR direct sequencing we confirmed that these dual gRNAs could specifically destroy HBV expressing template by removing the fragment between the cleavage sites of the two used gRNAs. Most importantly, gRNA-5 and gRNA-12 combination not only could efficiently suppressing HBsAg and/or HBeAg production, but also destroy the cccDNA reservoirs in HepAD38 cells. CONCLUSION: These results suggested that CRISPR/Cas9 system could efficiently destroy HBV expressing templates (genotypes A-D) without apparent cytotoxicity. It may be a potential approach for eradication of persistent HBV cccDNA in chronic HBV infection patients.

Heuristic hill-climbing search algorithm can do effectively pruning. In practice, it can be used to search a large hypothesis space to get an optimal or an approximate optimal solution. Beam search algorithm retains its advantage in efficiency while reducing the risk of converging to locally optimal hypotheses. Beam search algorithm is widely used in AI field. To k-size beam search, due to only k paths is maintained the key to optimize the accuracy of beam search is how to select the k paths. In most of search algorithms, the k candidates with the most high performance measure value are selected at each search step. In this paper, the author presented some methods of candidate selection of beam search approaches, and the thought of avoiding "full of blood brother nodes" is presented. The experiments were done on the UCI repository of machine learning databases

Tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a novel member of the TNF family and triggers apoptosis in a variety of malignant cells.We assessed the antitumour activity of TRAIL in six human pancreatic cancer cell lines by measuring apoptosis with sulforhodamine B cytotoxicity assays, DAPI staining and FACScans.All six cell lines were sensitive to TRAIL-induced apoptosis. The expression levels of TRAIL receptors R1, R2, R3 and R4 did not correlate with TRAIL sensitivity. Simultaneous treatment with TRAIL and the chemotherapeutic agent gemcitabine had a synergistic effect. Caspase-8 and -3 activities were increased by TRAIL treatment and apoptosis was largely blocked by caspase-8 and -3 inhibitors.Apoptotic cell death can be induced in pancreatic cancer cells by TRAIL treatment and this may be mediated through activation of caspases-8 and -3. Gemcitabine significantly increased the antitumour effect of TRAIL, suggesting novel therapeutic approaches.

Purpose: Bax plays an important role in the regulation of apoptosis induced by chemotherapy and other stimuli. We therefore investigated the role of Bax in drug-induced apoptosis in human pancreatic cancer cells. Method: A tetracycline-inducible retroviral expression vector bearing the human bax-α gene was constructed. ASPC-1 cells were stably infected with this vector. The sensitivity of the transformed cell line to gemcitabine and 5-Fu was assessed. Results: Western blots revealed that Bax expression was enhanced in these cells by the tetracycline analogue doxycycline. Enhanced expression of Bax itself did not inhibit the growth rate of infected cells and did not influence expression of Bcl-2 and Bcl-xL. However, it significantly increased the sensitivity of cells to the chemotherapeutic drugs gemcitabine and 5-Fu. These drugs also activated caspase-8 and caspase-3 by up to ninefold. Caspase activation and/or an imbalance in Bax and Bcl-2/Bcl-xL expression may be the reasons for the augmentation of cytotoxicity by these drugs. Conclusions: The findings suggest that enhanced Bax expression may have therapeutic application in enhancing the efficacy of chemotherapy in pancreatic cancers.

Wuniu early tea (Camellia sinensis) is an important beverage consumed in China. Up to date, a lot of methods for identifying and chemical analysing have been done. However, there is no report on the effects of Wuniu early tea on cytochrome P450 isozymes. Therefore, the present objective of our study was to evaluate the potential effects of Wuniu early tea on cytochrome P450 isozymes P2C9, P1A2, P2C19 and P2B6 in rats with a cocktail approach including, matching probe drugs of tolbutamide, phenacetin, omeprazole and bupropion. These four probe drugs were simultaneously administered to rats after repeated Wuniu early tea administration. The pharmacokinetics of the probes in the plasma was simultaneous determined by high-performance liquid chromatography-mass spectrometry. The t 1/2 and AUC (0-∞) of tolbutamide increased significantly and CL z decreased remarkably in test rats after repeated Wuniu early tea administration. However, the main pharmacokinetic parameters of the other three probe drugs were not significantly different between control and test rats. The findings in this study suggested that Wuniu early tea could inhibit cytochrome P2C9 while did not influence on cytochrome P1A2, cytochrome P2C19 and cytochrome P2B6.