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Background. Immunotherapy has been considered as a promising cancer treatment for hepatocellular carcinoma (HCC). However, due to the particular immune environment of the liver, identifying patients who could benefit from immunotherapy is critical in clinical practice. Methods. The pyroptosis gene expression database of 54 candidates from The Cancer Genome Atlas (TCGA) were collected to discover the critical prognostic-related pyroptosis genes. A novel pyroptosis gene model was established to calculate the risk score. Kaplan–Meier analysis and receiver operating characteristic curve (ROC) were used to verify its predictive ability. The International Cancer Genome Consortium (ICGC) data was collected as external validation data to verify the model’s accuracy. We employed multiple bioinformatics tools and algorithms to evaluate the tumor immune microenvironment (TIME) and the response to immunotherapy. Results. Our study found that most pyroptosis genes were expressed differently in normal and tumor tissues and that their expression was associated with the prognosis. Then, a precise four-pyroptosis gene model was generated. The one-year area under the curves (AUCs) among the training, internal, and external validation patients were 0.901, 0.727, and 0.671, respectively. An analysis of survival data revealed that individuals had a worse prognosis than patients with low risk. The analysis of TIME revealed that the low-risk group had more antitumor cells, fewer immunosuppressive cells, stronger immune function, less immune checkpoint gene expression, and better immunotherapy response than the high-risk group. Immunophenoscore (IPS) analysis also demonstrated that the low-risk score was related to superior immune checkpoint inhibitors therapy. Conclusion. A nomogram based on the four-pyroptosis gene signature was a novel tool to predict the effectiveness of immunotherapy for HCC. Therefore, individualized treatment targeting the pyroptosis genes may influence TIME and play an essential role in improving the prognosis in HCC patients.

Ovarian cancer (OC) is a malignant neoplasm that displays increased vascularization. Angiopoietin-like 4 (ANGPTL4) is a secreted glycoprotein that functions as a regulator of cell metabolism and angiogenesis and plays a critical role in tumorigenesis. However, the precise role of ANGPTL4 in the OC microenvironment, particularly its involvement in angiogenesis, has not been fully elucidated. Our study demonstrates a significant upregulation of ANGPTL4 in OC specimens and its strong association with unfavorable prognosis. RNA-seq analysis affirms that ANGPTL4 facilitates OC development by driving janus kinase 2- signal transducer and activator of transcription 3 (JAK2-STAT3) signaling pathway activation. The interaction between ANGPTL4 and endothelial cell-specific molecule 1 (ESM1) promotes ANGPTL4 binding to lipoprotein lipase (LPL), thereby resulting in reprogrammed lipid metabolism and the promotion of OC cell proliferation, migration, and invasion. In the OC microenvironment, ESM1 may interfere with the binding of ANGPTL4 to integrin and vascular-endothelial cadherin (VE-Cad), which leads to stabilization of vascular integrity and ultimately promotes angiogenesis. Collectively, our findings underscore that ANGPTL4 promotes OC development via JAK signaling and induces angiogenesis in the tumor microenvironment through its interaction with ESM1.

In the current study, via applying a hybrid-ligand method, a novel Cu(II)-based coordination polymer, formulated as [Cu2(OH)(bib)(bpt)]·H2O (1) (H3bpt = biphenyl-3,4′,5-tricarboxylic acid, bib = 1,4-bis(1-imidazolyl)benzene), has been successfully obtained by a isothermal method. Its treatment effect against postpartum hemorrhage was assessed in the bio-study. Firstly, through the model establishment and compound treatment, the prothrombin activity in the plasma was assessed by measuring the prothrombin time. Next, the levels of the factor XI and IX were also assessed by ELISA (enzyme linked immunosorbent assay detection kit). In addition to this, the prevention activity of the compound on the disseminated intravascular coagulation (DIC) during postpartum hemorrhage was also evaluated by measuring the inflammatory response of the inflammatory cytokines, such as IL-1β (interleukin-1β) and TNF-α (tumor necrosis factor-α). The inflammatory signaling relative expression was assessed via the real time RT-PCR (reverse transcription-polymerase chain reaction) as well. Above all, we can see that the compound has excellent treatment activity on DIC disease by inhibiting p53 and nf-κb relative expression in the endothelial cells.

OBJECTIVE: To study the efficacy and safety of domestic generic levetiracetam in replacement of brand-name levetiracetam in the treatment of children with epilepsy. METHODS: A retrospective analysis was performed on the medical data of 154 children with epilepsy who received domestic generic levetiracetam in the inpatient or outpatient service of Guangdong Provincial People's Hospital from May 2019 to December 2020. Domestic generic levetiracetam and brand-name levetiracetam were compared in terms of efficacy and safety. RESULTS: For these 154 children, the epilepsy control rate was 77.3% (119/154) at baseline. At 6 months after switching to domestic generic levetiracetam, the epilepsy control rate reached 83.8% (129/154), which showed a significant increase (P0.05). The incidence of refractory epilepsy in children with no response after switching treatment was significantly higher than that in children with response (P

Neuronal nicotinic acetylcholine receptors containing α6 subunits (α6(*)-nAChRs) show highly restricted distribution in midbrain neurons associated with pleasure, reward, and mood control, suggesting an important impact of α6(*)-nAChRs in modulating mesolimbic functions. However, the function and pharmacology of α6(*)-nAChRs remain poorly understood because of the lack of selective agonists for α6(*)-nAChRs and the challenging heterologous expression of functional α6(*)-nAChRs in mammalian cell lines. In particular, the α6 subunit is commonly co-expressed with α4(*)-nAChRs in the midbrain, which masks α6(*)-nAChR (without α4) function and pharmacology. In this study, we systematically profiled the pharmacology and function of α6(*)-nAChRs and compared these properties with those of α4β2 nAChRs expressed in the same cell line. Heterologously expressed human α6/α3 chimeric subunits (α6 N-terminal domain joined with α3 trans-membrane domains and intracellular loops) with β2 and β3 subunits in the human SH-EP1 cell line (α6(*)-nAChRs) were used. Patch-clamp whole-cell recordings were performed to measure these receptor-mediated currents. Functionally, the heterologously expressed α6(*)-nAChRs exhibited excellent function and showed distinct nicotine-induced current responses, such as kinetics, inward rectification and recovery from desensitization, compared with α4β2-nAChRs. Pharmacologically, α6(*)-nAChR was highly sensitive to the α6 subunit-selective antagonist α-conotoxin MII but had lower sensitivity to mecamylamine and dihydro-β-erythroidine. Nicotine and acetylcholine were found to be full agonists for α6(*)-nAChRs, whereas epibatidine and cytisine were determined to be partial agonists. Heterologously expressed α6(*)-nAChRs exhibited pharmacology and function distinct from those of α4β2-nAChRs, suggesting that α6(*)-nAChRs may mediate different cholinergic signals. Our α6(*)-nAChR expression system can be used as an excellent cell model for future investigations of α6(*)-nAChR function and pharmacology.

Two-dimensional transition metal dichalcogenide are attracting increasing attention in electrochemical sensing due to unique nanoconstruction and electronic properties. The present work reports the preparation of flower-like vanadium disulfide (VS 2 ) by a simple one-step hydrothermal process and its properties are characterized by X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Data from XRD, SEM and TEM demonstrate that the as-prepared VS 2 product has ordered nanosheets stacking flower-structure which is constructed with many irregular nanosheets as a petal-like structure with thickness of several nanometers. A novel electrochemical biosensor was constructed for the determination of 17β-estradiol by immobilizing aptamer on the surface of VS 2 nanoflowers and gold nanoparticles modified glassy carbon electrode. Differential pulse voltammetry is applied for the determination of 17β-estradiol and a good linear relationship between the peak current and the logarithm of 17β-estradiol concentration from 1.0 × 10 −11 to 1.0 × 10 −8 M with a detection limit of 1.0 × 10 −12 M (3σ/S). The aptamer sensor exhibits a high sensitivity and remarkable reproducible analytical performance, and was successfully applied for the determination of 17β-estradiol in urine samples with recoveries of 92.0–105.2%. Our attempt will extend the application of VS 2 nanoflowers to electrochemical biosensing and catalytic areas.

We report a simple strategy to prepare 2-dimensional cobalt sulfide by the one-step hydrothermal process. The cobalt sulfide sample has a microstructure of interconnected sheets. As the electrode material for supercapacitors, the layered cobalt sulfide film exhibits superior performances with high specific capacitances (1314 F g −1 at 3 A g −1 ) as well as excellent cycle life (less than 8.3% decrease in specific capacitance after 500 cycles at 3 A g −1 ), making it suitable for high-performance supercapacitor application.

Two–dimensional (2D) transition metal dichalcogenide nanosheet is attracting increasing attention in energy storage due to unique nanoconstruction and electronic properties. In this work, molybdenum disulfide (MoS 2 ) nanosheet was prepared by a simple hydrothermal method, and its properties were characterized by X–ray powder diffraction, X–ray photoelectron spectroscopy, scanning electron microscopy and transmission electron microscopy. The electrochemical performances of the product were evaluated by cyclic voltammogram, galvanostatic charge–discharge and electrochemical impedance spectroscopy. The MoS 2 nanosheet showed a specific capacitance of 129.2 F g −1 at a current density of 1 A g −1 . In addition, the MoS 2 nanosheet electrode showed good cycle property as an excellent electrode material for electrochemical capacitors and the specific capacitance had only a slight decrease after 500 cycles (retention of 85.1%). The excellent electrochemical performance was attributed to the unique morphology of MoS 2 nanosheets, which possessed large specific surface area, unique 2D nanostructures and low equivalent series resistance.

Hyperspectral images classification is one of the important methods to identify image information, which has great significance for feature identification, dynamic monitoring and thematic information extraction, etc. Unsupervised classification without prior knowledge is widely used in hyperspectral image classification. This article proposes a new hyperspectral images unsupervised classification algorithm based on harmonic analysis(HA), which is called the harmonic analysis classifer (HAC). First, the HAC algorithm counts the first harmonic component and draws the histogram, so it can determine the initial feature categories and the pixel of cluster centers according to the number and location of the peak. Then, the algorithm is to map the waveform information of pixels to be classified spectrum into the feature space made up of harmonic decomposition times, amplitude and phase, and the similar features can be gotten together in the feature space, these pixels will be classified according to the principle of minimum distance. Finally, the algorithm computes the Euclidean distance of these pixels between cluster center, and merges the initial classification by setting the distance threshold. so the HAC can achieve the purpose of hyperspectral images classification. The paper collects spectral curves of two feature categories, and obtains harmonic decomposition times, amplitude and phase after harmonic analysis, the distribution of HA components in the feature space verified the correctness of the HAC. While the HAC algorithm is applied to EO-1 satellite Hyperion hyperspectral image and obtains the results of classification. Comparing with the hyperspectral image classifying results of K-MEANS, ISODATA and HAC classifiers, the HAC, as a unsupervised classification method, is confirmed to have better application on hyperspectral image classification.

Currently, the World confronts with several major problems, including environment pollution and energy shortage. To utilize metal-contaminated soils safely and to solve the problem of shortage of farmland for bioenergy development, we have postulated a new strategy of cultivating energy plants in Cd-contaminated soils for bioenergy production, and this can also be combined with phytoremediation. Here, we focus on the advantage and feasibility of this approach by a review of recent developments in basic and applied research relevant. It is concluded that cultivation of energy plants in metal-contaminated land for bioenergy production is a high beneficial, environment-friendly technique that is also technically feasible. It might not only cover the shortages of phytoremediation and bioenergy production, but also makes the metal-contaminated land fully utilized and productive, and this is benefited for both agriculture and farmers.

A simple and unique strategy for preparation of large-compound vesicle (LCV)-encapsulated multiwalled carbon nanotubes (MWCNTs) has been developed, and this involves dispersion of MWCNTs in H-shaped copolymers solution in DMF and encapsulation of MWCNTs with LCVs formed from hydrolysis and polycondensation of Si(OCH3)3 groups in the amphiphilic H-shaped copolymers, (PTMSPMA)2PEG(PTMSPMA)2. This unique noncovalent approach is nondestructive, and the original structure of MWCNTs remains in the resultant MWCNTs/LCVs nanocomposites. The morphologies of nanocomposites LCVs/MWCNTs are controlled by the chain length ratio (NPTMSPMA/NPEG) of PTMSPMA to PEG. For the H-shaped copolymers with NPMSPMA/NPEG ≤ 1.7, they self-assembled to form LCVs with dense cavities in the presence of MWCNTs in a mixture of DMF/H2O. When this ratio was more than 2.0, the large-compound micelle-wrapped MWCNTs were produced. This approach is potentially useful for preparation of MWCNTs encapsulated with various morphologies of polymers. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 3669–3679, 2009

A series of well-defined θ-shaped copolymers composed of polystyrene (PS) and poly(e-caprolactone) (PCL) with controlled molecular weight and narrow molecular weight distribution have been successfully synthesized without any purification procedure by the combination of atom transfer radical polymerization (ATRP), ring-opening polymerization (ROP), and the “click” chemistry. The synthetic process involves two steps: (1) synthesis of AB2 miktoarm star copolymers, which contain one PCL chain terminated with two acetylene groups and two PS chains with two azido groups at their one end, (α,α′-diacetylene-PCL) (ω-azido-PS)2, by ROP, ATRP, and the terminal group transformation; (2) intramolecular cyclization of AB2 miktoarm star copolymers to produce well-defined pure θ-shaped copolymers using “click” chemistry under high dilution. The 1H NMR, FTIR, and gel permeation chromatography techniques were applied to characterize the chemical structures of the resultant intermediates and the target polymers. Their thermal behavior was investigated by DSC. The mobility decrease of PCL chain across PS ring in the theta-shaped copolymers restricts the crystallization ability of PCL segment. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 2620–2630, 2009

Well-defined hetero eight-shaped copolymers composed of polystyrene (PS) and poly(e-caprolactone) (PCL) with controlled molecular weight and narrow molecular weight distribution were successfully synthesized by the combination of ring-opening polymerization, ATRP, and “click” reaction. The synthetic procedure involves three steps: (1) preparation of a tetrafunctional PS and PCL star copolymer with two PS and two PCL arms using the tetrafunctional initiator bearing two hydroxyl groups and two bromo groups; (2) synthesis of tetrafunctional star copolymer, (α-acetylene-PCL)2(ω-azido-PS)2, by the transition of terminal hydroxyl and bromo groups to acetylene and azido groups through the reaction with 4-propargyloxybutanedioyl chloride and NaN3 respectively; (3) intramolecular cyclization reaction to produce the hetero eight-shaped copolymers using “click” chemistry under high dilution. The 1H NMR, FTIR, and gel permeation chromatography techniques were applied to characterize the chemical structures of the resulted intermediates and the target polymers. Their thermal behavior was investigated by DSC, and their crystallization behaviors of PCL were studied by polarized optical microscopy. The decrease in chain mobility of the eight-shaped copolymers restricts the crystallization of PCL and the crystallization rate of PCL is slower in comparison with their corresponding star precursors. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 6496–6508, 2008

Well-defined figure-of-eight-shaped (8-shaped) polystyrene (PS) with controlled molecular weight and narrow polydispersities has been prepared by the combination of atom transfer radical polymerization (ATRP) and click chemistry. The synthesis involves two steps: 1) Preparation of a linear tetrafunctional PS with two azido groups, one at each end of the polymer chain, and two acetylene groups at the middle of the chain. 2) Intramolecular cyclization of the linear tetrafunctional PS at a very low concentration by a click reaction to produce the 8-shaped polystyrenes. The resulting intermediates and the target polymers were characterized by 1 H NMR and FT-IR spectroscopy, and gel permeation chromatography. The glass transition temperatures (T g s) were determined by differential scanning calorimetry and it was found that the decrease in chain mobility by cyclization resulted in higher T g s for 8-shaped polystyrenes as compared to their corresponding precursors.

A series of well-defined centipede-like copolymers with poly(glycidyl methacrylate) (PGMA) as main chain and poly(L-lactide) (PLLA) and polystyrene (PSt) as side chains have been synthesized successfully by combination of ring-opening polymerization and atom transfer radical polymerization (ATRP). The synthetic process includes three steps. (1) Synthesis of PGMA via ATRP; (2) preparation of macroinitiator with one bromine group and a hydroxyl group at every GMA unit of PGMA; (3) ring-opening polymerization of LLA and ATRP of St to obtain the asymmetric centipede-like copolymers. The number–average degrees of polymerization of PGMA backbone, PLLA and PSt side chains were determined by 1H-NMR spectra, and the molecular weights of the resultant intermediates and centipede-like copolymers were measured by gel permeation chromatography. The molecular weight distributions were narrow and the molecular weights of both the backbone and the side chains were controllable. The thermal behavior of the centipede-like copolymers was investigated by differential scanning calorimeter. With the increase of PSt side chain length, the glass transition temperature of PLLA side chains shifted to high temperature, and crystallization ability of PLLA side chains became poor. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 5580–5591, 2008

Heavy oil (distillation temperature: 320-340 ºC) derived from the direct coal liquefaction process using Shengli coal were hydrotreated using sulfided Ni-Mo/Al2O3, Ni-W/Al2O3, and Ni-W/SiO2 catalysts respectively. The sulfided catalysts were characterized by BET, XRD, H2-TPR and NH3-TPD respectively. The evaluations of the hydrodenitrogenation (HDN) and hydrodearomatization (HDA) properties of heavy oil on the three catalysts were carried out at 400 ºC and 5.0 MPa initial H2 pressure. The W-based catalysts displayed better performances than Mo-based catalysts for the HDN and HDA reactions. Al2O3 supported catalysts were found to have higher catalytic activities than on SiO2 supported ones. The activities of sulfided catalysts were associated mainly with the nature of active sites, acidity, metal sulfide crystallite size and the amount of the reducible sulfur species of metal sulfide. Derivados de óleo pesado (temperatura de destilação: 320-340 ºC), obtidos a partir do processo de liquefação direta do carvão, usando carvão Shengli foram tratados com hidrogênio usando catalisadores sulfetados Ni-Mo/Al2O3, Ni-W/Al2O3 e Ni-W/SiO2 respectivamente. Os catalisadores sulfetados foram caracterizados por BET, XRD, H2-TPR e NH3-TPD, respectivamente. As avaliações das propriedades de hidrodenitrogenação (HDN) e hidrodearomatização (HDA) do óleo pesado nos três catalisadores foram realizadas a 400 ºC e 5,0 MPa de pressão inicial de H2. Os catalisadores contendo W mostraram melhores desempenhos do que os catalisadores de Mo para as reações de HDN e HDA. Os catalisadores suportados em Al2O3 apresentaram maior atividade catalítica do que aqueles suportados em SiO2. As atividades dos catalisadores sulfetados foram associadas principalmente à natureza dos sítios ativos, acidez, tamanho do cristalito e da quantidade de espécies redutíveis de enxofre do sulfeto metálico

N-n-Butyl haloperidol iodide (F(2)) is a novel compound derived from haloperidol. In our previous work, F(2) was found to be an L-type calcium channel blocker which played a protective role in rat heart ischemic-reperfusion injury in a dose-dependent manner. In the current study, we aimed to investigate the effects and some possible mechanisms of F(2) on calcium transients in hypoxic/ischemic rat cardiac myocytes.Calcium transients' images of rat cardiac myocytes were recorded during simulated hypoxia, using a confocal calcium imaging system. The amplitude, rising time from 25% to 75% (RT25-75), decay time from 75% to 25% (DT75-25) of calcium transients, and resting [Ca(2+)](i) were extracted from the images by self-coding programs. In this study, hypoxia produced a substantial increase in diastolic [Ca(2+)](i) and reduced the amplitude of calcium transients. Both RT25-75 and DT75-25 of Ca(2+) transients were significantly prolonged. And F(2) could reduce the increase in resting [Ca(2+)](i)and the prolongation of RT25-75 and DT75-25 of Ca(2+) transients during hypoxia. F(2) also inhibited the reduction in amplitude of calcium transients which was caused by 30-min hypoxia. The activity of SERCA2a (sarcoplasmic reticulum Ca(2+)-ATPase, determined by test kits) decreased after 30-min ischemia, and intravenous F(2) in rats could ameliorate the decreased activity of SERCA2a. The inward and outward currents of NCX (recorded by whole-cell patch-clamp analysis) were reduced during 10-min hypoxia, and F(2) further inhibited the outward currents of NCX during 10-min hypoxia. All these data of SERCA2a and NCX might be responsible for the changes in calcium transients during hypoxia.Our data suggest that F(2) reduced changes in calcium transients that caused by hypoxia/ischemia, which was regarded to be a protective role in calcium homeostasis of ventricular myocytes, probably via changing the function of SERCA2a.

N-n-butyl haloperidol (F(2)), a novel compound of quaternary ammonium salt derivatives of haloperidol, was reported to antagonize myocardial ischemia/reperfusion injuries. The antiarrhythmic potential and electrophysiological effects of F(2) on rat cardiac tissues were investigated.In Langendorff-perfused rat hearts, the ventricular arrhythmias were induced by left anterior descending coronary artery of rat heart ligated for 20 min before the release of the ligature. F(2) provided some inhibitive effects against ischemia- and reperfusion-induced ventricular arrhythmias. In His bundle electrogram and epicardial ECG recordings, the drug produced bradycardia, delayed the conduction through the atrioventricular node and prolonged the Wenckebach cycle length and atrioventricular nodal effective refractory period. In whole-cell patch-clamp study, F(2) primarily inhibited the L-type Ca2+ current (I(Ca,L)) (IC(50) = 0.17 microM) with tonic blocking properties and little use-dependence. And the drug also decreased the Na+ current (IC(50) = 77.5 microM), the transient outward K+ current (IC(50) = 20.4 microM), the steady-state outward K+ current (IC(50) = 56.2 microM) and the inward rectifier K+ current (IC(50) = 127.3 microM).F(2) may be a promising drug for the treatment of ischemic heart disease with cardiac arrhythmia.

His bundle electrogram (HBE) recording is an important method for the study of the atrioventricular conduction system. However, the current HBE recording methods in isolated animal hearts have some disadvantages, such as unstable recording due to the difficulty in fixing electrodes as a result of intense heart beat, the small amplitude of the His signal or the possibility to destroy the integrity of heart structure. To overcome these disadvantages, we designed and manufactured reliable, inexpensive and easy-made bipolar cannula electrodes, which combine the functions of Langendorff-perfusion aortic cannula and recording electrodes. With the cannula electrodes, the operation of HBE recording becomes easier and clearer; hence, more stable recordings can be obtained in isolated rat hearts.

Puerarin, a major isoflavonoid compound from the Chinese herb, Ge-gen (Pueraria lobata), has effective treatment on myocardial and cerebral ischemia, glaucoma and sudden deafness in clinical setting in China. Our present work showed that puerarin (50, 150, 450 microM) concentration-dependently inhibited phenylephrine or KCl-induced contraction only in endothelium-intact rat aortic rings. In Ca(2+)-free solution, the antivasoconstriction of puerarin on phenylephrine was totally deprived. N (G)-nitro-L-arginine methyl ester, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, indomethacin and the three K(+) channel blockers, glibenclamide, tetraethylammonium and Ba(2+) displayed significant inhibitory effects on the antivasoconstriction of puerarin. 8-bromo-cGMP significantly strengthened the action of puerarin. Puerarin (10-160 microM) concentration-dependently induced the NO production in the rat aortic cells. These findings suggested that the antivasoconstriction elicited by puerarin is endothelium-dependent. NO/NO-cGMP pathway, PGI(2) and the opening of K(+) channels sensitive to glibenclamide, tetraethylammonium, and Ba(2+), which might be triggered by the extracellular Ca(2+) influx in the endothelium, appear to contribute to the antivasoconstriction of puerarin.

To investigate the effect of L-arginine on expression of human FVIII gene.Plasmid pcDNA6/V5-HisA-BDDhF VIII containing B domain deleted human coagulant factor VIII cDNA (BDDhF VIII cDNA) was constructed and transfected into human umbilical vein endothelial cells (HUVEC). After 72 h incubation with L-arginine (final concentration was 10 mmol/L) , the supernatant was collected for determining the antigen and clotting activity of human FVIII (FVIII: Ag and FVIII: C ) with ELISA and one stage clotting assay respectively. HUVECs were harvested for detecting human FVIII mRNA by Northern blot analysis. The five functional domains of BDDhFVIII cDNA including A1, A2, A3, C1 and C2 were amplified with PCR and inserted into pcDNA6/V5-HisA to construct the expression plasmids pcDNA6/V5-Hi-sA-BDDhFVIII-A1, pcDNA6/V5-HisA-BDDhFVIII-A2, pcDNA6/V5-HisA-BDDhFVIII-A3, pcDNA6/V5-HisA-BDDhFVIII-C1 and pcDNA6/V5-HisA-BDDhFVIII-C2, respectively. HUVEC were transfected with the five plasmids respectively and incubated with L-arginine (at the final concentration of 10 mmol/L) for 72 h. Nucleoli were then isolated and underwent run-on assay.After 24 h incubation with L-arginine, FVIII: Ag and FVIII: C were increased markedly in the supernatant of HUVEC [FVIII: Ag was (146.08 +/- 4.78) ng/ ml, and FVIII: C (0.752 +/- 0.009) U/ml/10(6) cells x 24 h, while in control supernatant without L-arginine, FVIII: Ag was (34.66 +/- 3.98) ng/ml, and FVIII: C (0.171 +/- 0.006) U/ml/10(6) cell x 24 h, P0.01]. Northern blot analysis indicated that, after adding L-arginine, the transcription of human FVIII mRNA was intensified remarkably in HUVEC transfected with pcDNA6/V5-HisA-BDDhFVIII, but no any transcription in those transfected with pcDNA6/V5-HisA. Run-on assay demonstrated that with L-arginine induction, A1 and A2 domains transcription was increased obviously, while no change in A3, C1 and C2 domains transcription.L-arginine increases expression of human FVIII gene in HUVEC through enhancing its transcription, particularly, domain A1 and A2 within FVIII gene.

We have previously shown that N-n-butyl haloperidol iodide (F2) derived from haloperidol reduces ischemia/reperfusion-induced myocardial injury by blocking intracellular Ca2+ overload. This study tested the hypothesis that cardio-protection with F2 is associated with an attenuation in the expression of early growth response gene 1 (Egr-1). In an in vivo rat model of 60 min coronary occlusion followed by 180 min of reperfusion, treatment with F2 significantly reduced myocardial injury evidenced by the reduction in release of plasma creatine kinase, myocardial creatine kinase isoenzyme and lactate dehydrogenase. In cultured neonatal rat cardiomyocytes of hypoxia for 3 h and reoxygenation for 1 h, F2 treatment attenuated necrotic and apoptotic cell death, as demonstrated by electron microscopy. Concomitant with cardio-protection by F2, the increased expression levels of Egr-1 mRNA and protein were significantly reduced in myocardial tissue and cultured cardiomyocytes as detected by reverse transcription-polymerase chain reaction, immunohistochemistry and immunocytochemistry. In conclusion, these results suggest that the protective effect of F2 on ischemia/reperfusion- or hypoxia/reoxygenation-induced myocardial injury might be partly mediated by downregulating Egr-1 expression.

N-n-butyl haloperidol iodide (F2), a novel compound derived from haloperidol, was synthesized by our drugs research lab. The present study aims to evaluate the protective effects of F2 on myocardial ischemia-reperfusion injury in vivo, and to try to find the protective mechanism of F2. The animal model of myocardial ischemia-reperfusion injury was established by ligaturing rabbit's left ventricular branch of coronary artery for 40 min and removing the ligation later to reperfuse for 40 min. Different doses of F2 were intravenously injected before the onset of ischemia. The changes of hemodynamics were recorded during the experiment, and the activities of superoxide dismutase (SOD), creatine kinase (CK), Ca2+-ATPase, Na+,K+-ATPase and the level of malondialdehyde (MDA) of myocardial tissue were detected after reperfusion. Administration of F2 could dose-dependently ameliorate the hemodynamics of ischemia-reperfusion injured myocardium. During the course of reperfusion, MAP, LVSP, +/-dP/dt(max) in all F2 groups were obviously higher than those in the ischemia-reperfusion control group, and LVEDP were lower. F2 could also reduce the production of MDA, and maintain the activities of SOD, Ca2+-ATPase, Na+,K+-ATPase, and minimize the leakage of CK out of myocardial cells in a dose-dependent manner. These results suggested that F2 had apparent protective effects against myocardial ischemia-reperfusion injury.

To observe the effects of Kupffer cells on hepatic drug metabolic enzymes.Kunming mice were i.p. injected with GdCl310, 20, 40 mg/kg to decrease the number and block the function of kupffer cells selectively. The contents of drug metabolic enzymes, cytochrome P450, NADPH-cytochrom C redutase (NADPH-C), aniline hydroxylase (ANH), aminopyrine N-demethylase (AMD), erythromycin N-demethylase (EMD), and glutathione s-transferase (mGST) in hepatic microsome and S9-GSTpi, S9-GST in supernatant of 9 000 g were accessed 1 d after the injection. The time course of alteration of drug metabolic enzymes was observed on d 1, 3, and 6 treated with a single dose GdCl3. Mice were treated with Angelica sinensis polysaccharides (ASP) of 30, 60, 120 mg/kg, i.g., qd x 6 d, respectively and the same assays were performed.P450 content and NADPH-C, ANH, AMD, and EMD activities were obviously reduced 1 d after Kupffer cell blockade. However, mGST and S9-GST activities were significantly increased. But no relationship was observed between GdCl3 dosage and enzyme activities. With single dose GdCl3 treatment, P450 content, NADPH-C, and ANH activities were further decreased following Kupffer cell blockade lasted for 6 d, by 35.7%, 50.3%, 36.5% after 3 d, and 57.9%, 57.9%, 63.2% after 6 d, respectively. On the contrary, AMD, EMD, mGST, and S9-GST activities were raised by 36.5%, 71.9%, 23.1%, 35.7% after 3 d, and 155%, 182%, 21.5%, 33.7% after 6 d, respectively. Furthermore, the activities of drug metabolic enzymes were markedly increased after 30 mg/kg ASP treatment, and decreased significantly after 120 mg/kg ASP treatment. No change in activity of S9-GSTpi was observed in the present study.Kupffer cells play an important role in the modulation of drug metabolic enzymes. The changes of drug metabolic enzyme activities depend on the time of kupffer cell blockade and on the degree of Kupffer cells activated. A low concentration of ASP increases the activities of drug metabolic enzymes, but a high concentration of ASP decreases the activities of drug metabolic enzymes.

AIM: To study the modulating effect of GdCl3 and Angelica Sinensis polysaccharides (ASP) on differentially expressed genes in liver of hepatic immunological mice by cDNA microarray. METHODS: Hepatic immunological injury was induced by lipopolysaccharide (LPS ip, 0.2 mg·kg-1) in bacillus calmetteguerin (BCG ip, 1 mg·kg-1) primed mice; A single dose of 20 mg·kg-1 GdCl3 was simultaneously pretreated and 30 mg·kg-1 ASP (ig, qd × 7 d) was administrated when the BCG+LPS was primed. The mice were sacrificed at the end of the 7th day after ip LPS for 6 h and the liver was removed quickly. The PCR products of 512 genes were spotted onto a chemical material-coated glass plate in array. The DNAs were fixed to the glass plate after series of treatments. The total RNAs were isolated from the liver tissue, and were purified to mRNAs by Oligotex. Both mRNAs from the normal liver tissue and the liver tissue from the mice with hepatic immunological injury or that pretreated with GdCl3 or ASP were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for fluorescent signals and showed differences between the two tissues. RESULTS: Among the 512 target genes, 18 differed in liver tissue of hepatic immunological injury mice, and 6 differed in those pretreated by ASP, 7 differed in those pretreated by GdCl3. CONCLUSION: cDNA microarray technique is effective in screening the differentially expressed genes between two different kinds of tissue. Further analysis of those obtained genes will be helpful to understand the molecular mechanism of hepatic immunological injury and to study the intervention of drug. Both ASP and GdCl3 can decrease the number of the differentially expressed genes in liver tissue of mice with hepatic immunological injury.