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Grain boundary management is critical to the performance and stability of polycrystalline perovskite solar cells (PSCs), especially large-area devices. However, typical passivators are insulating in nature and limit carrier transport. Here, we design a supramolecular binder for grain boundaries to simultaneously passivate defects and promote hole transport across perovskite grain boundaries. By doping the monoamine porphyrins (MPs, M = Co, Ni, Cu, Zn, or H) into perovskite films, MPs self-assemble into supramolecules at grain boundaries. Organic cations in perovskites protonate MPs in supramolecules to form ammonium porphyrins bound on the perovskite grain surface, to passivate defects and extract holes from the perovskite lattice. Periodic polarons in supramolecules (especially NiP-supramolecule) promote the transport of extracted holes across boundaries, reducing nonradiative carrier recombination. The NiP-doped PSCs reveal a certified efficiency of 22.1% for an active area of 1.0 cm2 with the remarkably improved open-circuit voltage and fill factor. The unencapsulated device retained over 80% initial performance under AM 1.5G solar light continuous illumination or heating at 85 °C over 3000 h.

The role of hepatitis E virus (HEV) in developing hepatocellular carcinoma (HCC) is unclear. Our study aimed to investigate the role of HE infection in HCC development and the effect of hepatitis B virus (HBV) and HEV coinfection on HCC risk. A hospital-based case-control study was conducted. A total of 474 eligible HCC cases and 586 control patients were successfully recruited. The fasting venous blood was collected from the patients at the first visited to hospital and HBV infection and HEV infection were examined within 5 days. Crude and adjusted odd ratios (ORs) with 95% confidence interval (95% CI) were estimated by using logistic regression model. HBV infection (OR: 63.10, 95% CI: 42.02-97.26) rather than HEV infection (OR: 1.08, 95% CI: 0.721-1.65) was associated with an increased risk of HCC after adjustment for confounders. The association between HBV infection and HCC risk was more remarkable in male (OR: 72.61, 95% CI: 45.10-121.38) than in female (OR: 61.89, 95% CI: 25.74-169.26). In comparison with patients who infected with neither HEV nor HBV, those who infected with only HBV (OR: 69.62, 95% CI: 40.90-123.52) and who coinfected with HEV and HBV (OR: 67.48, 95% CI:37.23-128.19) were significantly associated with an increased risk after adjustment for potential confounders. The results showed that HBV infection rather than HEV infection was associated with an increased risk of HCC, and the HEV infection may alleviate the promoting impact of HBV on HCC development.

It took only 11 years for the power conversion efficiency (PCE) of perovskite solar cells (PSCs) to increase from 3.8% to 25.2%. It is worth noting that, as a new thin-film solar cell technique, defect passivation at the interface is crucial for the PSCs. Decorating and passivating the interface between the perovskite and electron transport layer (ETL) is an effective way to suppress the recombination of carriers at the interface and improve the PCE of the device. In this work, several acceptor–donor–acceptor (A–D–A) type fused-ring organic semiconductors (FROS) with indacenodithiophene (IDT) or indacenodithienothiophene (IDDT) as the bridging donor moiety and 1,3-diethyl-2-thiobarbituric or 1,1-dicyromethylene-3-indanone as the strong electron-withdrawing units, were deposited on the SnO2 ETL to prepare efficient planar junction PSCs. The PCEs of the PSCs increased from 18.63% for the control device to 19.37%, 19.75%, and 19.32% after modification at the interface by three FROSs. Furthermore, impedance spectroscopy, steady-state and time-resolved photoluminescence spectra elucidated that the interface decorated by FROSs enhance not only the extraction of electrons but also the charge transportation at the interface between the perovskite and ETL. These results can provide significant insights in improving the perovskite/ETL interface and the photovoltaic performance of PSCs.

BACKGROUND Hepatitis E virus (HEV) superinfection is a suspected promoting factor for hepatocellular carcinoma (HCC) in patients with chronic hepatitis and cirrhosis. However, to date, very few cases of HEV-related HCC have been reported. Nevertheless, the role of HEV re-infection in cirrhotic liver without other chronic hepatitis infections has rarely been explored. CASE SUMMARY A 53-year-old male farmer was diagnosed with liver cirrhosis and splenomegaly in August 2016, accompanied with negative HEV-IgM and positive HEV-IgG. No evidence of hepatitis B virus or hepatitis C virus infection was found. Since then the patient was evaluated for liver function and viral parameters every 3 mo. In June 2017, the patient presented severe fatigue with whole body itching and was diagnosed with HCC. Afterwards this patient experienced quick HCC development, progression, relapse, and metastasis in the following 8 mo, and presented persistent dual positivity of HEV-IgM and HEV-IgG. This patient had a long history of smoking and alcohol consumption. CONCLUSION This unique case invokes the importance of HEV surveillance and treatment among cirrhotic patients, HCC cases, and blood donors.

The defects on the surface of low-temperature-processed electronic transport layers hindered the development of efficient flexible perovskite solar cells. Herein, we develop a universal NdCl3 dosin...

Owing to excellent performance and dopability, spiro-OMeTAD remains an irreplaceable hole transporting material (HTM) in perovskite solar cells (PSCs). In order to further improve the performance of spiro-OMeTAD based PSCs, a Lewis base can be introduced into the structure of spiro-OMeTAD wisely, which can keep the advantages of spiro-OMeTAD while incorporating the functionality of a Lewis base in passivating the surface of the perovskite. Therefore, spiro-type HTMs (spiro-CN-OMeTAD with a dicyano group and spiro-PS-OMeTAD with a thiocarbonyl group) were synthesized and confirmed by density functional theory (DFT) calculations and X-ray single-crystallographic diffraction. Spiro-CN-OMeTAD as an HTM is certified to have a suitable interfacial band alignment with the perovskite, good film quality and effective defect passivation, which facilitate the resulting device to achieve an efficiency of 19.90% with a high open-circuit voltage, low hysteresis, and improved stability. This study provides an alternative strategy for the molecular design of better HTMs in high-performance PSCs.

Antibiotics, as veterinary drugs, have made extremely important contributions to disease prevention and treatment in the animal breeding industry. However, the accumulation of antibiotics in animal food due to their overuse during animal feeding is a frequent occurrence, which in turn would cause serious harm to public health when they are consumed by humans. Antibiotic residues in food have become one of the central issues in global food safety. As a safety measure, rapid and effective analytical approaches for detecting these residues must be implemented to prevent contaminated products from reaching the consumers. Traditional analytical methods, such as liquid chromatography, liquid chromatography mass spectrometry, and capillary electrophoresis, involve time-consuming sample preparation and complicated operation and require expensive instrumentation. By comparison, surface-enhanced Raman spectroscopy (SERS) has excellent sensitivity and remarkably enhanced target recognition. Thus, SERS has become a promising alternative analytical method for detecting antibiotic residues, as it can provide an ultrasensitive fingerprint spectrum for the rapid and noninvasive detection of trace analytes. In this study, we comprehensively review the recent progress and advances that have been achieved in the use of SERS in antibiotic residue detection. We introduce and discuss the basic principles of SERS. We then present the prospects and challenges in the use of SERS in the detection of antibiotics in food. Finally, we summarize and discuss the current problems and future trends in the detection of antibiotics in food.

Background Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae has become a public health concern. This study aimed to compare the clinical outcomes of patients with nonurinary source bacteraemia caused by ESBL-producing Escherichia coli (E. coli) or Klebsiella pneumoniae (ESBL-producing EK) receiving β-lactam/β-lactamase inhibitor combinations (BLICs) versus carbapenem treatment and assess the risk factors of mortality with these two drugs. Methods We conducted a retrospective single-centre study of adult hospitalised patients with ESBL-producing EK bloodstream infection (BSI) from nonurinary source at our centre over a 4-year period. One hundred and eighty patients who received BLICs or carbapenems were included in the analysis. The outcome variables were 14-day treatment failure and 30-day mortality. For more reliable results, propensity score analysis was performed to compare the efficacy of the two drugs and analyse their risk factors for 30-day mortality. Results Out of 180 patients, 114 received BLICs, and 66 received carbapenem therapy. Compared to carbapenem-treated patients, those treated with BLICs were older and had higher age-adjusted Charlson comorbidity index, but they had shorter stay in the hospital. Additionally, their Pitt bacteraemia score, SOFA score, rate of leukaemia, and immune compromise were lower. After propensity score matching (PSM), the baseline characteristics of patients in the two treatment groups were balanced. BLICs were associated with a higher 14-day treatment failure rate (20.6%, 13/63) than carbapenems (16.3%, 7/43), although the difference was not significant in either univariate analysis (P = 0.429) or multivariate analysis (P = 0.122). And the 30-day mortality rate in BTG (11.1%, 7/63) and CTG (11.6%, 5/43) did not significantly differ (univariate analysis, P = 0.926; multivariate analysis, P = 0.420). In the multivariate analysis, after PSM, leukaemia was the only independent predictor of mortality in both BTG and CTG. Conclusions Our study showed that BLICs had higher 14-day treatment failure rate compared with carbapenems, although there were no statistically significant differences because of the small number of patients, therefore, further evaluation of the efficacy of BLICs is needed.

We have demonstrated a two-step wet-chemical route to prepare dense a SnO2 blocking layer (BL) for boosting efficaciously the optoelectronic performance of mesoporous-TiO2 based perovskite solar cells (PSCs). Through SnCl4 water-bath for moderate time, the surface features of SnO2 BL are availably optimized and recombination loss of the devices is perceptibly suppressed. The influence of bath-time upon the microstructures and photoelectronic properties of SnO2 BL and photovoltaic characteristics of the corresponding PSCs, has been discussed detailedly. Deep-going explorations on charge dynamics confirm that SnCl4 bath would facilitate charge transport, hinder charge accumulation and inhibit nonradiative recombination. Upon the basis of surface decoration of SnO2, we improve the open-circuit photovoltage from 1.02 V of initial cell to 1.06 V, and achieve a maximum photovoltaic efficiency of 17.14% and null hysteresis. Accordingly, the device with large area (1.0 cm2) based on modified SnO2 have higher PCE of 14.35% and weaker hysteresis index of 0.019 than its unmodified SnO2 courtpart.

Thromboxane A2 (TXA2) has been implicated in the pathogenesis of diabetic vascular complications, although the underlying mechanism remains unclear. The present study investigated the alterations in TXA2 receptor signal transduction in type 2 diabetic renal arteries. The contraction of renal arterial rings in control (db/m+) mice and type 2 diabetic (db/db) mice was measured by a Multi Myograph System. Intracellular calcium concentration ([Ca2+]i) in vascular smooth muscle cells was measured by Fluo-4/AM dye and confocal laser scanning microscopy. Quantitative real-time PCR and Western blot analysis were used to determine gene and protein expression levels, respectively. A stable TXA2 mimic U46619 caused markedly stronger dose-dependent contractions in the renal arteries of db/db mice than in those of db/m+ mice. This response was completely blocked by a TXA2 receptor antagonist GR32191 and significantly inhibited by U73122. U46619-induced vasoconstriction was increased in the presence of nifedipine in db/db mice compared with that in db/m+ mice, whereas the response to U46619 did not differ between the two groups in the presence of SKF96365. Sarcoplasmic reticulum Ca2+ release-mediated and CaCl2-induced contractions did not differ between the two groups. In db/db mice, store-operated Ca2+(SOC) entry-mediated contraction in the renal arteries and SOC entry-mediated Ca2+ influx in smooth muscle cells were significantly increased. And the gene and protein expressions of TXA2 receptors, Orai1 and Stim1 were upregulated in the diabetic renal arteries. Therefore the enhancement of U46619-induced contraction was mediated by the upregulation of TXA2 receptors and downstream signaling in the diabetic renal arteries.

Summary Effect of ZnO nanorod surface on fabricating the perovskite solar cell and its performance were studied. Varied thickness of ZnO nanorod arrays with rough surface condition were achieved through the control of hydrothermal growth time and acid treatment. Samples based on modified ZnO nanorod arrays exhibit an impressive increase on the open-circuit voltage (Voc) and fill factor (FF) compared with the untreated ones. Further research onto the surface topography and electrochemical impedance spectroscopy test indicates that the improvement should be attributed to the suppression of the charge recombination rate at the ZnO nanorod/CH3NH3PbI3 interface. In order to enhance the short-circuit current density (Jsc) performance one step further, Cl-doped perovskite crystal was introduced into the cell. Because of its longer electron diffusion length, an impressive Jsc is received. The final combination of the two methods with the optimized thickness of the ZnO nanorod brought a total power conversion efficiency of 13.3% together with Voc~0.92 V, Jsc~23.1 mA/cm2, and FF~63%. This work highlights the importance of the surface morphology of the electron transport layer and its interface contact with the light absorbing layer in a solar cell structure. Copyright © 2017 John Wiley & Sons, Ltd.

The role of statins in reducing the incidence of contrast-induced acute kidney injury (CI-AKI) remains controversial. We sought to evaluate the association between CI-AKI and high plasma exposure of statins in coronary artery disease (CAD) patients undergoing coronary angiography (CAG). This association was first evaluated in 1,219 patients with CAD receiving atorvastatin (AT) therapy and validated in 635 patients receiving rosuvastatin (RST) therapy. The plasma concentrations of statins were quantified using validated UPLC-MS/MS methods and CI-AKI incidence was assessed during the first 48 h postoperatively. Among all participants (n = 1,854), AKI occurred in 57 of 1219 (4.7%) in the AT cohort and 30 of 635 (4.7%) in the RST cohort. High plasma AT-all exposure was associated with increased risk of CI-AKI (odds ratio [OR]: 2.265; 95% confidence interval [CI]: 1.609–3.187; p < 0.0001). Plasma AT-all concentration in the CI-AKI group (22.40 ± 24.63 ng/mL) was 2.6-fold higher than that in the control group (8.60 ± 9.65 ng/mL). High plasma RST exposure also significantly increased the risk of CI-AKI (OR: 2.281; 95% CI: 1.441–3.612; p = 0.0004). We further divided patients into two subgroups for each statin according to baseline renal function, and association between high plasma statin exposure and CI-AKI still remained highly significant in both subgroups. This study suggests for the first time that high plasma exposure of statins may significantly increase the risk of CI-AKI. Statins should be used with greater caution in CAD patients undergoing CAG to reduce the occurrence of CI-AKI.

Power loss is a serious problem for all power companies. To find effective means of abnormal electricity iidentification is a popular research field in recent years. This paper puts forward an anomalous electricity use model based on multi-dimensional compound features of electricity users. The support vector machine, local outlier factor, correlation measurement based on the similar user power load, and correlation change rate measurement based on the most relevant users—these four algorithms are adopted to extract four-dimensional compound features of anomalous electricity use from the perspective of global anomaly, local anomaly, regional space, and time sequence. Next, the logistic regression (LR) model is trained based on the compound features. After training, the LR model is adopted as the final anomalous electricity use identification model. Analysis of the practical power load data of users suggests that the LR model combine respective advantages of the four-dimensional compound features. Detection of anomalies using the LR model is an effective approach, which can reliably and accurately identify residents’ anomalous electricity use. From the accuracy rate, recall rate, precision rate and scores of F1, it can be seen that the LR model is significantly superior to SVM.

Objective This study was designed to evaluate the pharmacokinetics (PK) and safety of eptifibatide in healthy Chinese volunteers and provide information for the further study in the Chinese population. Methods 30 healthy volunteers (15 male) were enrolled in the study and divided into three dose groups (45 µg x kg⁻¹, 90 µg x kg⁻¹, and 180 µg x kg⁻¹). Plasma and urine samples were drawn after one single-bolus administration and measured by LC-MS/MS. The plasma and urine data were analyzed simultaneously by the population approach using the NONMEM software and evaluated by the visual predicted check (VPC) and bootstraping. The PK profiles of dose regimens approved for a Western population in the Chinese population were simulated. Results A two-compartment model adequately described the PK profiles of eptifibatide. The clearance (CL) and the distribution volume (V₁) of the central compartment were 0.128 L x h⁻¹ x kg⁻¹ and 0.175 L x kg⁻¹, respectively. The clearance (Q) and V₂of the peripheral compartment were 0.0988 L x h⁻¹ x kg⁻¹ and 0.147 L x kg⁻¹, respectively. The elimination fraction from plasma to urine (F₀) was 17.2%. No covariates were found to have a significant effect. Inter-individual variabilites were all within 33.9%. The VPC plots and bootstrap results indicated good precision and prediction of the model. The simulations of the approved regimens in the Chinese population showed much lower steady-state concentrations than the target concentration obtained from the Western clinical trials. No severe safety events were found in this study. Conclusions The PK model of eptifibatide was established and could provide PK information for further studies in the Chinese population.

Cyclins/retinoblastoma protein (pRb) pathway participates in cardiomyocyte hypertrophy. MicroRNAs (miRNAs), the endogenous small non-coding RNAs, were recognized to play significant roles in cardiac hypertrophy. But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy. This study investigates the potential role of microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy. An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC), and in a mouse with transverse aortic constriction (TAC) and in a mouse with subcutaneous injection of phenylephrine (PE) respectively. In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte and based on Ang-II-induced neonatal mouse ventricular cardiomyocyte respectively. We demonstrated that miR-16 expression was markedly decreased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats and mice. Overexpression of miR-16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes, and inhibition of miR-16 induced a hypertrophic phenotype in cardiomyocytes. Expressions of cyclins D1, D2 and E1, and the phosphorylated pRb were increased in hypertrophic myocardium and hypertrophic cardiomyocytes, but could be reversed by enforced expression of miR-16. Cyclins D1, D2 and E1, not pRb, were further validated to be modulated post-transcriptionally by miR-16. In addition, the signal transducer and activator of transcription-3 and c-Myc were activated during myocardial hypertrophy, and inhibitions of them prevented miR-16 attenuation. Therefore, attenuation of miR-16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2 and E1, and activating cyclin/Rb pathway, revealing that miR-16 might be a target to manage cardiac hypertrophy.

A photoanode with Ga-doped ZnO nanorods has been prepared on F-doped SnO2 (FTO) coated glass substrate and its application in dye-sensitized solar cells (DSSCs) has been investigated. Ga-doped ZnO nanorods have been synthesized by an electric-field-assisted wet chemical approach at 80?C. Under a direct current electric field, the nanorods predominantly grow on cathodes. The results of the X-ray photoelectron spectroscopy and photoluminescence verify that Ga dopant is successfully incorporated into the ZnO wurtzite lattice structure. Finally, employing Ga-doped ZnO nanorods with the length of ~5 μm as the photoanode of DSSCs, an overall energy conversion efficiency of 2.56% is achieved. The dramatically improved performance of Ga-doped ZnO based DSSCs compared with that of pure ZnO is due to the higher electron conductivity.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a desmosomal disease. Desmosomes and gap junctions are important structural components of cardiac intercalated discs. The proteins plakophilin-2 (PKP-2) and connexin43 (Cx43) are components of desmosomes and gap junctions, respectively. This study was conducted to determine whether Cx43 expression is affected by the mutation of the PKP-2 gene in patients with ARVC. A novel mutation was detected in a typical patient with ARVC. The mutated gene was transfected into rat mesenchymal stem cells expressing Cx43 through a pReversied-M-29 plasmid. Cx43 expression was detected using quantitative polymerase chain reaction analysis. Cx43 expression was significantly decreased in the mutant PKP-2 group compared with that in the wild-type PKP-2 group. In conclusion, PKP-2 affected Cx43 expression at the gene transcription level in the patient with ARVC.

The molecular mechanisms underlying anthracyclines-induced cardiotoxicity have not been well elucidated. MiRNAs were revealed dysregulated in the myocardium and plasma of rats received Dox treatment. MicroRNA-34a-5p (miR-34a-5p) was verified increased in the myocardium and plasma of Dox-treated rats, but was reversed in rats received Dox plus DEX treatments. Human miR-34a-5p was also observed increased in the plasma of patients with diffuse large B-cell lymphoma after 9- and 16-week epirubicin therapy. Up-regulation of miR-34a-5p was observed in Dox-induced rat cardiomyocyte H9c2 cells. MiR-34a-5p could augment Bax expression, but inhibited Bcl-2 expression, along with the increases of the activated caspase-3 and mitochondrial potentials in H9C2 cells. MiR-34a-5p was verified to modulate Sirt1 expression post-transcriptionally. In parallel to Sirt1 siRNA, miR-34a-5p could enhance p66shc expression, accompanied by increases of Bax and the activated caspase-3 and a decrease of Bcl-2 in H9c2 cells. Moreover, enforced expression of Sirt1 alleviated Dox-induced apoptosis of H9c2 cells, with suppressing levels of p66shc, Bax, the activated caspase-3 and miR-34a-5p, and enhancing Bcl-2 expression. Therefore, miR-34a-5p enhances cardiomyocyte apoptosis by targeting Sirt1, activation of miR-34a-5p/Sirt1/p66shc pathway contributes to Dox-induced cardiotoxicity, and blockage of this pathway represents a potential cardioprotective effect against anthracyclines.

Storage and transportation of natural gas with NGH method is a kind of novel technology developing rapidly at domestic and overseas currently. The stability and dissociation characteristic of hydrate at different temperature and pressure is a significant basis on its application and development. The existence of anomalous self-preservation effect indicates that gas hydrates have favorable stability in special low temperature range, and it provides theoretical and experimental basis for storage and transportation of natural gas with NGH method. The paper reviews study history and research development of hydrate self-preservation effect, and points out challenges for developing storage and transportation technology of natural gas with NGH.

Macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, plays an important role in the pathogenesis of atrial fibrillation; however, the upstream regulation of MIF in atrial myocytes remains unclear. In the present study, we investigated whether and how MIF is regulated in response to the renin-angiotensin system and oxidative stress in atrium myocytes (HL-1 cells). MIF protein and mRNA levels in HL-1 cells were assayed using immunofluorescence, real-time PCR, and Western blot. The result indicated that MIF was expressed in the cytoplasm of HL-1 cells. Hydrogen peroxide (H2O2), but not angiotensin II, stimulated MIF expression in HL-1 cells. H2O2-induced MIF protein and gene levels increased in a dose-dependent manner and were completely abolished in the presence of catalase. H2O2-induced MIF production was completely inhibited by tyrosine kinase inhibitors genistein and PP1, as well as by protein kinase C (PKC) inhibitor GF109203X, suggesting that redox-sensitive MIF production is mediated through tyrosine kinase and PKC-dependent mechanisms in HL-1 cells. These results suggest that MIF is upregulated by HL-1 cells in response to redox stress, probably by the activation of Src and PKC.

Aims Antiplatelet treatment can cause a change in plasma levels of platelet microRNAs (miRNAs). However, it is not clear whether the plasma level of platelet miRNAs can predict clinical outcomes of antiplatelet treatment. The present study aimed to evaluate the association of plasma miR-16, miR%E2%80%9121, miR-126, miR-26b, and miR-223 with the risk of clinical outcomes in dual antiplatelet-treated patients after percutaneous coronary intervention (PCI). Methods and results A total of 491 Han Chinese patients who had received PCI and dual antiplatelet therapy were sequentially recruited to the study and followed for up to one year. Plasma concentrations of five candidate miRNAs early the next morning after PCI were determined by quantitative reverse transcription PCR. The effect of the plasma miRNA level on major adverse cardiovascular events (MACE) within one year and bleeding within six months were assessed. We found that a higher plasma miR-126 level was significantly associated with a higher risk in terms of time-to-MACE. When compared with the plasma miR-126 level in the first three quartiles, the hazard ratio (HR) for the plasma miR-126 level in the fourth quartile was 2.61 (95% CI: 1.32-5.18, p=0.006). Multivariable Cox regression analysis showed that diabetes mellitus, ejection fraction, hypertension and a higher plasma miR-126 level were independent risk factors for MACE. Plasma miR-223 level was not an independent predictive marker for MACE. There was no significant association between the level of five plasma miRNAs and bleeding events during six-month follow-up. Conclusions Based on these results, we suggest that plasma miR-126 could be a potential marker for predicting major adverse cardiac events in patients after PCI.

// Wen-Si Zhu 1,2,* , Chun-Mei Tang 2,3,* , Zhen Xiao 1,2,* , Jie-Ning Zhu 1,2 , Qiu-Xiong Lin 1,2 , Yong-Heng Fu 1,2 , Zhi-Qin Hu 2,3 , Zhuo Zhang 2,4 , Min Yang 1,2 , Xi-Long Zheng 5 , Shu-Lin Wu 1,2 and Zhi-Xin Shan 1,2 1 Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangzhou, China 2 Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China 3 Southern Medical University, Guangzhou, China 4 School of Medicine, South China University of Technology, Guangzhou, China 5 The Libin Cardiovascular Institute of Alberta, Department of Biochemistry & Molecular Biology, The University of Calgary, Calgary, Canada * These authors have contributed equally to this work Correspondence to: Zhi-Xin Shan, email: // Keywords : microRNA-214-3p, cardiac fibrosis, cardiac myofibroblast, EZH1, EZH2, Pathology Section Received : August 31, 2016 Accepted : October 28, 2016 Published : November 03, 2016 Abstract The role of microRNA-214-3p (miR-214-3p) in cardiac fibrosis was not well illustrated. The present study aimed to investigate the expression and potential target of miR-214-3p in angiotensin II (Ang-II)-induced cardiac fibrosis. MiR-214-3p was markedly decreased in the fibrotic myocardium of a mouse Ang-II infusion model, but was upregulated in Ang-II-treated mouse myofibroblasts. Cardiac fibrosis was shown attenuated in Ang-II-infused mice received tail vein injection of miR-214-3p agomir. Consistently, miR-214-3p inhibited the expression of Col1a1 and Col3a1 in mouse myofibroblasts in vitro . MiR-214-3p could bind the 3’-UTRs of enhancer of zeste homolog 1 (EZH1) and -2, and suppressed EZH1 and -2 expressions at the transcriptional level. Functionally, miR-214-3p mimic, in parallel to EZH1 siRNA and EZH2 siRNA, could enhance peroxisome proliferator-activated receptor-γ (PPAR-γ) expression and inhibited the expression of Col1a1 and Col3a1 in myofibroblasts. In addition, enforced expression of EZH1 and -2, and knockdown of PPAR-γ resulted in the increase of Col1a1 and Col3a1 in myofibroblasts. Moreover, the NF-κB signal pathway was verified to mediate Ang-II-induced miR-214-3p expression in myofibroblasts. Taken together, our results revealed that EZH1 and -2 were novel targets of miR-214-3p, and miR-214-3p might be one potential miRNA for the prevention of cardiac fibrosis.

The role of microRNA-1 (miR-1) in ischemia/reperfusion (I/R)-induced injury is not well illustrated. The present study aimed to investigate the expression and potential target of miR-1 in the myocardium of a rat model of I/R. The apoptosis of cardiomyocytes in the ischemic rat myocardium increased on day 1, then attenuated on day 3 and day 7 post-I/R. Heat shot protein 90 (Hsp90) aa1 mRNA expression was decreased post-I/R and Hsp90aa1 protein level was decreased on day1 post-I/R, but was reversed on day 3 and day 7 post-I/R. MiR-1 was downregulated post-I/R and repression of miR-1 in cultured neonatal rat ventricular cells (NRVCs) led to an increase of Bcl-2 and decreases of Bax and active caspase-3. Dual luciferase reporter assays revealed that miR-1 interacted with the 310–315 nt site at the 3′UTR of Hsp90aa1 and miR-1 was verified to inhibit Hsp90aa1 expression at the posttranscriptional level. Over-expression of Hsp90aa1 could attenuate oxygen-glucose deprivation (OGD)-induced apoptosis of NRVCs. Additionally, miR-1 mimic, in parallel to Hsp90aa1 siRNA, could enhance OGD-induced apoptosis of NRVCs. Taken together, our results reveal that Hsp90aa1 is a novel target of miR-1 and repression of miR-1 may contribute to the recovery of Hsp90aa1 during myocardial I/R.

Background In the early stage of diabetes, the cardiac ejection fraction is preserved, despite the existence of the subclinical cardiac dysfunction to some extent. However, the detailed phenotype of this dysfunction and the underlying mechanism remain unclear. To improve our understanding of this issue, we used low-dose STZ and high-fat diet to induce type 2 diabetic models in rats. The effects and the mechanism associated with the early stages of the disease were analyzed. Methods The type 2 diabetic mellitus (T2DM) in SD rats were induced through 30 mg/kg STZ and high-fat diet. Two-dimensional spackle-tracking echocardiography (STE) and the dobutamine test were performed to examine the cardiac function. Calcium transients of left ventricular myocytes were detected and the related intracellular signalling factors were analyzed by western blotting. Results After 6-weeks, T2DM rats in left ventricular (LV) diastole showed decreased global and segment strain(S) levels (P

The T-type Ca(2+) current (I(Ca,T)) plays an important role in the pathogenesis of atrial fibrillation (AF). The present study sought to investigate the role of macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, in the regulation of T-type Ca(2+) channels (TCCs) in atrial myocytes. We used the whole-cell voltage-clamp technique and biochemical assays to study the regulation and expression of I(Ca,T) in atrial myocytes. Gene levels of the α1G and α1H subunit of TCCs were decreased in human atrial tissue of patients with AF. In cultured atrium-derived myocytes (HL-1 cells), mouse recombinant MIF (20 or 40 nm, 24 h) suppressed peak I(Ca,T) in a concentration-dependent manner, impaired the voltage-dependent activation of I(Ca,T) and downregulated TCC α1G and α1H mRNA. The Src inhibitors genistein and PP1 significantly enhanced I(Ca,T). The reduction of I(Ca,T) and TCC subunit mRNA induced by recombinant MIF could be reversed by genistein and PP1. The TCC α1G associated with Src in HL-1 cells and mouse cardiomycytes. Macrophage migration inhibitory factor is involved in the pathogenesis of AF, probably by decreasing the T-type calcium current in atrium-derived myocytes through impairment of channel function and activation of c-Src kinases, representing a potential pathogenic mechanism in atrial fibrillation.

MicroRNA-1 (miR-1) is approved involved in cardiac hypertrophy, but the underlying molecular mechanisms of miR-1 in cardiac hypertrophy are not well elucidated. The present study aimed to investigate the potential role of miR-1 in modulating CDKs-Rb pathway during cardiomyocyte hypertrophy. A rat model of hypertrophy was established with abdominal aortic constriction, and a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocytes (NRVCs). We demonstrated that miR-1 expression was markedly decreased in hypertrophic myocardium and hypertrophic cardiomyocytes. Dual luciferase reporter assays revealed that miR-1 interacted with the 3'UTR of CDK6, and miR-1 was verified to inhibit CDK6 expression at the posttranscriptional level. CDK6 protein expression was observed increased in hypertrophic myocardium and hypertrophic cardiomyocytes. Morover, miR-1 mimic, in parallel to CDK6 siRNA, could inhibit PE-induced hypertrophy of NRVCs, with decreases in cell size, newly transcribed RNA, expressions of ANF and β-MHC, and the phosphorylated pRb. Taken together, our results reveal that derepression of CDK6 and activation of Rb pathway contributes to the effect of attenuation of miR-1 on provoking cardiomyocyte hypertrophy.

1. It has been reported that allitridi, an active compound extracted from garlic, has many cardiovascular effects. However, it remains unknown whether allitridi affects major repolarization currents, such as the transient outward K(+) current (I(to) ), ultrarapid delayed rectifier K(+) current (I(Kur)) and the L-type Ca(2+) current (I(Ca)), in human atrial myocytes. 2. In the present study, we investigated the effects of allitridi on I(to), I(Kur), I(Ca) and the action potential in human isolated atrial myocytes using the whole-cell patch recording technique. 3. Allitridi reversibly inhibited I(to), but not I(Kur) and I(Ca), in human atrial myocytes. These effects of allitridi on I(to) were concentration dependent (IC(50) = 44.9 μmol/L). Inactivation of I(to) was accelerated and the voltage-dependent inactivation potential was shifted towards the negative direction. Allitridi (30 μmol/L) significantly prolonged action potential duration in human atrial myocytes. 4. The results of the present study indicate that allitridi inhibits I(to), but not I(Kur) and I(Ca), and prolongs the action potential duration in human atrial myocytes.

We sought to find blood-based biomarkers that can be used to predict efficacy in advanced non-small cell lung cancer patients treated with bevacizumab plus chemotherapy. Blood was collected before treatment and after 6 weeks of therapy from patients who were participating in a phase 4 trial. Plasma vascular endothelial growth factor (VEGF) levels were evaluated by ELISA. A total of eight single nucleotide polymorphisms in four candidate genes were analyzed by PCR and sequencing. A total of 45 patients enrolled in a clinical trial at Guangdong General Hospital between August 2007 and March 2008 were used as subjects. The median survival times of OS was 25.6 and 13.4 months in the low and high groups, respectively, when the median posttreatment plasma VEGF level (46.63 pg/ml) was used as the cut-off point (P = 0.0284). Patients carrying the AA genotype at the −6C > A polymorphism in laminin 5 (LN5) were more likely to exhibit reduced hemoglobin compared with patients carrying the CA/CC genotype (OR = 8.364, χ2 = 5.34, P = 0.021). Similar associations were found at the −89A > G and −260C > A polymorphisms in LN5. Patients with the CC genotype at the −6C > A polymorphism in LN5 had an increased risk of neutropenia than those with the CA/AA genotype (OR = 4.444, χ2 = 5.116, P = 0.030). Our results show improved survival in patients with lower posttreatment plasma VEGF levels treated with bevacizumab plus chemotherapy; thus, the posttreatment plasma VEGF level may be a promising biomarker to predict clinical benefit early in the course of therapy. Polymorphisms in LN5 were associated with a reduced level of hemoglobin and neutropenia.

Background and Purpose— MIF has been implicated to function in many inflammatory processes. This study examined whether MIF expression was affected in stroke and its underlying molecular mechanism. Methods— ELISA and qRT-PCR were used to detect MIF protein and mRNA in PBMCs from stroke patients, the ischemic rat brains, and controls. A MIF promoter assay under hypoxia was performed. Results— MIF protein and mRNA were significantly increased in stroke patients. Increasing levels of MIF were correlated to the severity of stroke and peaked 24 hours after stroke. MIF was significantly upregulated in focal ischemic rat brains. The activity of the human MIF promoter was significantly increased under hypoxia compared to normoxia. Conclusions— MIF gene expression is upregulated after stroke, and hypoxia signaling plays an important role in upregulation of MIF expression under stroke.

Peripheral circulating lymphocytes are easily accessible cells for investigating changes in beta-adrenergic receptors (ADRBs) in humans, but previous reports indicate that these cells only express ADRB2. This study aimed to investigate whether ADRB1 and ADRB3 were expressed in peripheral lymphocytes and the changes of ADRBs in congestive heart failure. Our study demonstrates that ADRB1, ADRB2, and ADRB3 coexist in human peripheral lymphocytes, with differential binding property and expression level. Patients with congestive heart failure had significantly decreased total ADRB density and mRNA levels of ADRB1 and ADRB2 genes, but not ADRB3, compared with healthy subjects. The levels of mRNA of ADRB1 and ADRB2 in peripheral lymphocytes from patients with congestive heart disease were significantly increased after drug treatment. Our study, for the first time, indicates that human peripheral lymphocytes coexpress ADRB1, ADRB2, and ADRB3, which has important implications for precisely predicting clinical response to drug therapy in congestive heart failure.

Aim: To study the multiple dose clinical pharmacokinetics of risperidone and its main active metabolite, 9-hydroxyrisperidone, in Chinese female patients with schizophrenia. Methods: The subjects were 23 Chinese female inpatients aged 18−65 years who met the CCMD-III (third revision of the Chinese Criteria of Mental Disorders) criteria for schizophrenia. Subjects were tested after 17 d of treatment with 2 mg risperidone twice daily. Plasma concentrations of risperidone and 9-hydroxy-risperidone were assayed by using validated high performance liquid chromatography-mass spectrometry (HPLC-MS) methods. Results: Risperidone was rapidly absorbed ( T max was 1.6 h) and its T1/2 in plasma was short (3.2 h). 9-hydroxy-risperidone was quickly metabolized from the parent drug with a mean T max of 2.5 h. It had a long half-life of 24.7 h. The C ss av of risperidone and 9-hydroxyrisperidone were 36.9±33.1 and 110.6±30.5 μg·h·L -1 , respectively, and the AUC ss 0–12 were 443.2±397.4 and 1327.2±402.3 μg·h·L-1, respectively. CL/F and V/F of risperidone were 8.7±6.2 L/h and 34.1±24.3 L, respectively. Interindividual variations for pharmacokinetic parameters were quite large for risperidone. All 23 subjects experienced high prolactin levels when treated with risperidone. However there was no correlation between prolactin level and the concentration of risperidone, 9-hydroxy-risperidone, or the active moiety. Conclusion: Risperidone showed large interindividual variations in pharmacokinetics. Administration of risperidone resulted in high serum prolactin levels. The results indicate that systemic exposure to risperidone and 9-hydroxy-risperidone in female Chinese schizophrenic patients is higher relative to published data for white Caucasian patients. Larger studies regarding the PK/PD relationship may be required to develop a reasonable clinical dosage regimen for Chinese female patients.

MicroRNAs play important roles in myocardial fibrosis during diabetic cardiomyopathy. The present study aimed to investigate the role of microRNA-208b (miR-208b) in diabetic myocardial fibrosis. Compared to the db/m control mice, Smad3 was activated , fibrosis-related genes expression were significantly up-regulated, and miRNAs were dysregulated in the myocardium of the 16-week-old diabetic db/db mice. miR-208b was confirmed up-regulated in the db/db diabetic myocardium in vivo and in mouse cardiomyocytes and cardiac fibroblasts after treatment with Ang-II, TGF-β and high glucose/glucose oxidase (HG/Go) in vitro , respectively. By using recombinant adenovirus expressing CD63-GFP, rAd-CD63-GFP, we infected neonatal mouse cardiomyocytes with rAd-CD63-GFP and tracked the secreted exosomes with GFP. We found that miR-208b was increased in the secreted exosomes from HG/Go-treated cardiomyocytes, and the exosomes with increased miR-208b could enhance fibrosis associated Col1a1, α-SMA and CTGF expressions in cardiac fibroblasts. Transfection of miR-208b mimic could increase Col1a1, α-SMA and CTGF expression in a dose-dependent manner in cardiac fibroblasts. However, blockage of miR-208b could inhibit fibrosis related genes expression. Mtf2 and Pgrmc1 were verified modulated by miR-208b at post transcriptional level in vitro . Consistently, Mtf2 and Pgrmc1 expressions were decreased in the diabetic myocardium, and knockdown of Mtf2 or Pgrmc1could increase fibrosis related genes expression in cardiac fibroblasts. Smad3 inhibitor, Naringenin, could dramatically inhibit miR-208b expression in cardiac fibroblasts. Taken together, we demonstrated that miR-208b was up-regulated in diabetic fibrotic myocardium, Mtf2 and Pgrmc1 mediated the effect of miR-208b on enhancing Col1a1, α-SMA and CTGF expression in diabetic myocardial fibrosis.

OBJECTIVE: To test the hypothesis that locally directed gene transfer of human vascular endothelial growth factor 165 (hVEGF165) is able to passivate hepatic venous metallic stents by accelerating endo­thelialization and augmenting the biocompatibility of endovascular stents. METHODS: Complexes of pAdtrackCMV-hVEGF165 and lipofectamine were smeared homogeneously on the surface of the stents coated with poly-L-lysine. Bare stents were used as controls. All stents were implanted into the right hepatic vein by transjugular intrahepatic venous stent deployment. RESULTS: At the end of the first week after implantation, green fluorescence and the expression of the hVEGF165 gene were detected in the transferred stent vessels of the treated group, but were not detected in the control group. Scanning electron microscopy revealed that the endothelialization of stents was more pronounced in the treated group than that in the control group. At the end of the eighth week after implantation, quantitative angiography analysis showed the internal diameter of the stent was significantly greater in the treated group than in the control group. The de novo intima, their mean areas and percentage cross-sectional area narrowing in the treated group were significantly less than those in the control group. Immunohistochemical analysis indicated that the proliferation of vascular smooth muscle cells was more active in the control group than that in the treated group. CONCLUSION: Local gene transfer of hVEGF165 can passivate endovascular stents by accelerating stent endothelialization and enhancing their biocompatibility in the hepatic vein, resulting in a reduction of thrombus formation and attenuation of intimal hyperplasia.

Background: The feasibility of hands-free transthoracic continuous determination of pulmonary artery (PA) diastolic pressure (PAD) and cardiac output (CO) by Doppler ultrasound has not been previously demonstrated. We developed a 2.5-MHz spherical transducer mounted in an external housing to permit steering in 360 degrees (Contison). The external housing was attached to the chest wall using an adhesive patch. Methods and Results: Fifty patients in the coronary care department who had PA catheters had Doppler ultrasound studies. The 2.5-MHz spherical transducer was placed at the left sternal border to permit imaging of the pulmonic valve and was attached to a commercial ultrasound machine. The PA was imaged and its diameter measured. The pulmonary flow velocity signal was recorded and the time velocity integral obtained. The CO was calculated as: CO=time velocity integral of the PA systolic flow velocity signal × π diameter 2 divided by 4 × heart rate. The pulmonary regurgitation signal was then recorded and the end-diastolic velocity of the regurgitant signal was measured. Right atrial pressure was assessed from the jugular venous pressure or from the size and pulsatility of the inferior vena cava. The PADP was calculated as: PADP=4 end-diastolic velocity of the regurgitant signal 2 + right atrial pressure. The CO, PADP, and pulmonary wedge pressure were recorded from the PA catheter immediately after the ultrasound studies. Serial data were obtained every half hour or 1 hour up to a maximum of 5 hours. Adequate Doppler signals were obtained in 43 patients. Results: There was a good correlation between the PADP by Doppler versus PA catheter ( r = 0.90, standard error of the estimate=3.3 mm Hg); PADP by Doppler versus PA wedge pressure ( r = 0.88, standard error of the estimate=3.7 mm Hg); and CO by Doppler versus PA catheter ( r = 0.92, standard error of the estimate=0.7 L/min). Conclusion: The 2.5-MHz spherical transducer permitted accurate assessment of CO and PAD. This transducer could be of potential value in monitoring patients in the intensive care setting. (J Am Soc Echocardiogr 2002;15:1381-6.)

Percutaneous coronary intervention (PCI) is widely used in clinical treatment of coronary artery disease. However, the effects of PCI on preventing restenosis after revascularization and improving the quality of life were not satisfying. Huxin Formula is formulated by modifying an experienced Chinese medicine formula and has been widely used in clinical practice due to its marked effects on coronary heart disease. A multicentre double-blind randomized controlled clinical trial was designed to evaluate the effects and safety of Huxin Formula in patients undergoing PCI. Our results showed that there was no significant difference between the two groups in main outcomes. For patients with ejection fraction (EF)>50%, score of the quality of life scale was higher in treatment group compared with control group. For patients with unstable angina, score of the quality of life scale in 360 days was significantly higher in treatment group compared with control group (P<0.05). No obvious adverse reaction was found in the use of Huxin Formula. In conclusion, Huxin Formula, believed to be a safe treatment for patients after PCI, has benefits in improving the quality of life in patients with unstable angina though it failed to show superiority in primary and secondary outcomes.

Circular RNAs (circRNAs) participate in regulating gene expression in diverse biological and pathological processes. The present study aimed to investigate the mechanism underlying the modulation of circRNA_000203 on expressions of fibrosis-associated genes in cardiac fibroblasts. Masson trichrome

Carvedilol, a nonselective β-adrenoreceptor antagonist, protects against myocardial injury induced by acute myocardium infarction (AMI). The mechanisms underlying the anti-fibrotic effects of carvedilol are unknown. Recent studies have revealed the critical role of microRNAs (miRNAs) in a variety of cardiovascular diseases. This study investigated whether miR-29b is involved in the cardioprotective effect of carvedilol against AMI-induced myocardial fibrosis. Male SD rats were randomized into several groups: the sham surgery control, left anterior descending (LAD) surgery-AMI model, AMI plus low-dose carvedilol treatment (1 mg/kg per day, CAR-L), AMI plus medium-dose carvedilol treatment (5 mg/kg per day, CAR-M) and AMI plus high-dose carvedilol treatment (10 mg/kg per day, CAR-H). Cardiac remodeling and impaired heart function were observed 4 weeks after LAD surgery treatment; the observed cardiac remodeling, decreased ejection fraction, and fractional shortening were rescued in the CAR-M and CAR-H groups. The upregulated expression of Col1a1, Col3a1, and α-SMA mRNA was significantly reduced in the CAR-M and CAR-H groups. Moreover, the downregulated miR-29b was elevated in the CAR-M and CAR-H groups. The in vitro study showed that Col1a1, Col3a1, and α-SMA were downregulated and miR-29b was upregulated by carvedilol in a dose-dependent manner in rat cardiac fibroblasts. Inhibition of ROS-induced Smad3 activation by carvedilol resulted in downregulation of Col1a1, Col3a1, and α-SMA and upregulation of miR-29b derived from the miR-29b-2 precursor. Enforced expression of miR-29b significantly suppressed Col1a1, Col3a1, and α-SMA expression. Taken together, we found that smad3 inactivation and miR-29b upregulation contributed to the cardioprotective activity of carvedilol against AMI-induced myocardial fibrosis.

KRAS mutation plays an important role in the pathogenesis of pancreatic cancer. However, the role of wild-type KRAS in the progression of pancreatic cancer remains unknown. The present study was to investigate the expression of the Ras GTPase activating protein (DAB2IP) in pancreatic cancer and its clinical significance.The expression of DAB2IP in pancreatic cancer cell lines and normal human pancreatic ductal epithelial cells was analyzed by Western blotting and real-time quantitative reverse transcription-PCR (qRT-PCR). The KRAS mutational types of pancreatic cancer tissues obtained from pancreatic cancer patients (n=20) were also analyzed. Subsequently, DAB2IP expression was detected in pancreatic cancer tissues, adjacent and normal pancreatic tissues (n=2) by immunohistochemistry, and the relationship between DAB2IP expression and the clinical characteristics of patients was evaluated.Western blotting and qRT-PCR results showed that DAB2IP expression in pancreatic cancer cells with wild-type KRAS was lower than that in those with mutation-type KRAS and normal human pancreatic ductal epithelial cells (P0.05). Immunohistochemistry showed that DAB2IP expression was lower in pancreatic cancer tissues than that in adjacent and normal pancreatic tissues (Z=-4.000, P=0.000). DAB2IP expression was lower in pancreatic cancer patients with the wild-type KRAS gene than that in those with KRAS mutations (WilcoxonW=35.000, P=0.042). Furthermore, DAB2IP expression in patients with perineurial invasion was lower than that in those without invasion (WilcoxonW=71.500, P=0.028). DAB2IP expression was lower in patients with more advanced stage than that in those with early clinical stage (WilcoxonW=54.000, P=0.002).DAB2IP expression was reduced in patients with pancreatic cancer compared with those with no cancer. DAB2IP expression was correlated with the KRAS gene, perineurial invasion and clinical stage of the disease. Our data indicated that DAP2IP expression can be used as a potential prognostic indicator and a promising molecular target for therapeutic intervention in patients with pancreatic cancer.

Background : The renin–angiotensin system (RAS) is regarded as one of the most important regulatory systems for cardiovascular homeostasis. In this study, we investigated associations of the five genetic polymorphisms in the RAS and presence of coronary artery disease (CAD) in type 2 diabetes. Methods : Genotyping was performed by polymerase chain reaction–restriction fragment length polymorphism analysis. We used the generalised multifactor dimensionality reduction (GMDR) method to identify gene–gene interactions. Results : The D allele in the ACE gene was significantly more frequent in type 2 diabetic patients with CAD ( p = 0.04). In multivariate logistic regression analysis, the DD genotype was associated with a significantly increased risk of CAD ( p = 0.044). 1675G/A variant in the AT2R gene was found to be associated with CAD in female subjects with type 2 diabetes ( p = 0.025). The three other polymorphisms of the RAS do not seem to influence the development of CAD in type 2 diabetes. No significant gene–gene interaction for any combinations of genotypes was found in the GMDR method. Conclusion : The DD variant of the ACE gene polymorphism is associated with increased risk of developing CAD in Chinese patients with type 2 diabetes. A slight impact of AT2R 1675G/A polymorphism on CAD was found only in female diabetic patients.

Laminin 5 (Ln5) is an extracellular matrix protein that plays an important role in cell migration and tumor invasion. This study explored the expression of Ln5 and the role of its relationships with PTEN, phospho-EGFR (p-EGFR) and phospho-Akt (p-Akt) in the prognosis of patients with non-small cell lung cancer (NSCLC). The protein expression of Ln5, PTEN, p-EGFR and p-Akt was assessed by immunohistochemical analysis, and their relationships to prognosis were analyzed. Protein expression of Ln5, p-EGFR and p-Akt was detected in 61.2 (60/98), 60.2 (59/98) and 45.3% (43/95) of patients with NSCLC, respectively. Loss of PTEN expression was found in 67.7% of tumors (65/96). Ln5 expression was related to patient gender, histology and p-Akt expression (χ(2)=3.901, 4.549 and 6.985, respectively; P=0.048, 0.033 and 0.008, respectively). Patients with positive Ln5 expression had marginally poorer survival than Ln5-negative patients (median survival time 56.4 months vs. not reached; χ(2)=3.346; P=0.067). Overall survival was significantly different in patients with positive Ln5 expression combined with loss of PTEN, positive p-EGFR expression or positive p-Akt expression. Cox regression analysis showed that stage, co-expression of Ln5 and p-Akt, and PTEN were the three most independent prognostic factors for patients with NSCLC (χ(2)=27.906; P0.0005). The results highlight the complex relationships between extracellular matrix proteins and key signaling pathway molecules in tumorigenesis. Changes in the expression of Ln5 plus PTEN, p-EGFR or p-Akt define a distinct subset of lung cancers. Patients with such cancers have poorer survival and require early treatment that impacts survival.

Compared with genetic factors, drug interactions are largely unexplored in pharmacogenetic studies. This study sought to systematically investigate the effects of VKORC1, STX4A, CYP2C9, CYP4F2, CYP3A4, and GGCX gene polymorphisms and interacting drugs on warfarin maintenance dose.A retrospective study of 845 Chinese patients after heart valve replacement receiving long-term warfarin maintenance therapy was conducted. Thirteen polymorphisms in the six genes were genotyped, and 36 drugs that may interact with warfarin were investigated.Single-nucleotide polymorphism association analysis showed that VKORC1, CYP2C9 and CYP4F2 variations were highly associated with the warfarin maintenance dose. Among 36 drugs that may interact with warfarin, fluconazole, amiodarone, and omeprazole were associated with the requirement for 45.8, 16.7, and 16.7% lower median warfarin dose (all P0.05 with a false discovery rate0.05). The final pharmacogenetic equation explained 43.65% of interindividual variation of warfarin maintenance dose with age, body surface area, VKORC1 g.3588GA, CYP2C9*3, CYP4F2 c.1297GA, amiodarone, fluconazole, and diltiazem accounting for 1.97, 2.74, 24.12, 3.94, 1.64, 5.92, 2.47, and 0.84% of variation.The present study indicated that VKORC1, CYP4F2, and CYP2C9 genotypes and interacting drugs had a significant impact on the warfarin maintenance dose in Chinese patients with heart valve replacement and demonstrated that integrating interacting drugs can largely improve the predictability of the dose algorithm.

Aim of this study The panax notoginseng saponins (PNS) have been clinically used for the treatment of cardiovascular diseases and stroke in China. Evidences demonstrated that PNS could protect cardiomyocytes from injury induced by ischemia, but the underlying molecular mechanisms of this protective effect are still unclear. This study was aimed to investigate the protective effect and potential molecular mechanisms of PNS on apoptosis in H9c2 cells in vitro and rat myocardial ischemia injury model in vivo. Materials and methods H9c2 cells subjected to serum, glucose and oxygen deprivation (SGOD) were used as in vitro models and SD rats subjected to left anterior descending (LAD) coronary artery ligation were used as in vivo models. The anti-apoptotic effect of PNS was evaluated by Annexin V/PI analysis or TUNEL assay. Mitochondrial membrane potential (Δψm) was detected by JC-1 analysis. The expression of Akt and phosphorylated Akt (p-Akt) were detected by western blot assay. Results PNS exhibited anti-apoptotic effect both in H9c2 cells and in ischemic myocardial tissues. However, the effect was blocked in vitro by LY294002, a specific PI3K inhibitor. The anti-apoptotic effect of PNS was mediated by stabilizing Δψm in H9c2 cells. Furthermore the indices of the left ventricular ejection fractions (EF), left ventricular fractional shortening (FS), left ventricular dimensions at end diastole (LVDd) and left ventricular dimensions at end systole (LVDs) suggested that PNS improved rats cardiac function. PNS significantly increased p-Akt both in H9c2 cells and in ischemic myocardial tissues and this effect was also blocked by LY294002 in H9c2 cells. Conclusion Results of this study suggested that PNS could protect myocardial cells from apoptosis induced by ischemia in both the in vitro and in vivo models through activating PI3K/Akt signaling pathway.

Background Diabetes has been regarded as an inflammatory condition which is associated with left ventricular diastolic dysfunction (LVDD). The purpose of this study was to examine the expression levels of macrophage migration inhibitory factor (MIF) and G protein-coupled receptor kinase 2 (GRK2) in patients with early diabetic cardiomyopathy, and to investigate the mechanisms involved in MIF expression and GRK2 activation. Methods 83 patients in the age range of 30-64 years with type 2 diabetes and 30 matched healthy men were recruited. Left ventricular diastolic function was evaluated by cardiac Doppler echocardiography. Plasma MIF levels were determined by ELISA. To confirm the clinical observation, we also studied MIF expression in prediabetic rats with impaired glucose tolerance (IGT) and relationship between MIF and GRK2 expression in H9C2 cardiomyoblasts exposed to high glucose. Results Compared with healthy subjects, patients with diabetes have significantly increased levels of plasma MIF which was further increased in diabetic patients with Left ventricular diastolic dysfunction (LVDD). The increased plasma MIF levels in diabetic patients correlated with plasma glucose, glycosylated hemoglobin and urine albumin levels. We observed a significant number of TUNEL-positive cells in the myocardium of IGT-rats but not in the control rats. Moreover, we found higher MIF expression in the heart of IGT with cardiac dysfunction compared to that of the controls. In H9C2 cardiomyoblast cells, MIF and GRK2 expression was significantly increased in a glucose concentration-dependant manner. Furthermore, GRK2 expression was abolished by siRNA knockdown of MIF and by the inhibition of CXCR4 in H9C2 cells. Conclusions Our findings indicate that hyperglycemia is a causal factor for increased levels of pro-inflammatory cytokine MIF which plays a role in the development of cardiomyopathy occurring in patients with type 2 diabetes. The elevated levels of MIF are associated with cardiac dysfunction in diabetic patients, and the MIF effects are mediated by GRK2.

We investigated whether transplantation of bone marrow mesenchymal stem cells (BMSC) with induced BMSC (iBMSC) or uninduced BMSC (uBMSC) into the myocardium could improve the performance of post-infarcted rat hearts. BMSCs were specified by flowcytometry. IBMSCs were cocultured with rat cardiomyocyte before transplantation. Cells were injected into borders of cardiac scar tissue 1 week after experimental infarction. Cardiac performance was evaluated by echocardiography at 1, 2, and 4 weeks after cellular or PBS injection. Langendorff working-heart and histological studies were performed 4 weeks after treatment. Myogenesis was detected by quantitative PCR and immunofluorescence. Echocardiography showed a nearly normal ejection fraction (EF) in iBMSC-treated rats and all sham control rats but a lower EF in all PBS-treated animals. The iBMSC-treated heart, assessed by echocardiography, improved fractional shortening compared with PBS-treated hearts. The coronary flow (CF) was decreased obviously in PBS and uBMSC-treated groups, but recovered in iBMSC-treated heart at 4 weeks (P < 0.01). Immunofluorescent microscopy revealed co-localization of Superparamagnetic iron oxide (SPIO)-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. These data provide strong evidence that iBMSC implantation is of more potential to improve infarcted cardiac performance than uBMSC treatment. It will open new promising therapeutic opportunities for patients with post-infarction heart failure.

In clinical trials, the epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) administered concomitantly with first-line cytotoxicity chemotherapy failed to confer a survival benefit to patients with non-small-cell lung cancer (NSCLC). The aim of this study was to test whether paclitaxel followed by gefitinib is superior to other treatment schedules of NSCLC cell lines and to clarify the underlying mechanisms. Human lung cancer cell lines with wild-type and mutant-type EGFR genes were used as in vitro models to define the differential effects of various schedules of paclitaxel with gefitinib treatment on cell growth, signaling pathway, and cell cycle distribution. Sequence-dependent antiproliferative effects differed between EGFR-TKI-resistant and EGFR-TKI-sensitive lung cancer cell lines. Exposure to paclitaxel resulted in an increased pEGFR level. This increase in phosphorylation was inhibited by subsequent exposure to gefitinib, whereas during the reverse sequence, the inhibition effect was reduced. After paclitaxel exposure, a higher level of pEGFR was observed in mitotic than in interphase cells. The sequence of paclitaxel followed by gefitinib resulted in greater anti-VEGF activity than did the reverse sequence. We confirmed that gefitinib arrested cells in G1, and paclitaxel arrested them in S phase. The sequence of paclitaxel followed by gefitinib arrested cells in G1, whereas the reverse sequence arrested cells in S and G2 phases. These findings suggest that the sequence of paclitaxel followed by gefitinib may be superior to other sequences in treating NSCLC cell lines. Our results also provide molecular evidence to support clinical treatment strategies for patients with lung cancer.

To depict the interaction between atrial fibrillation (AF) and inflammatory reaction, studies were taken to measure the activity of NF-kappaB in myocardium, the concentration of regional inflammatory factors and the pathological process of the right atrium in patients with AF.Patients with valvular disease with AF or sinus rhythm (SR) were recruited and compared. Before the extracorporal circulation, about 250 mg tissue of right atrium was collected for pathological examination. The activity of NF-kappaB in myocardium was measured by electrophoretic mobility shift assay (EMSA), and the concentration of cardiac tissue interleukin-6 (IL-6) and tumor necrosis factor (TNF-alpha) was determined by radioimmuoassay.Patients with valvular disease with AF exhibited higher NF-kappaB activity, higher concentration of TNF-alpha and IL-6, severe lymphomonocyte infiltration, and more fibrosis than those patients with valvular disease with SR. There were significant positive correlations among NF-kappaB activity and levels of TNF-alpha and IL-6 and collagen volume fraction.This study proved the presence of inflammation in atrial myocardium by triggering inflammatory reaction.