In Operando Identification of In Situ Formed Metalloid Zincδ+ Active Sites for Highly Efficient Electrocatalyzed Carbon Dioxide Reduction.

Electrochemical CO2 -to-CO conversion provides a possible way to address problems associated with the greenhouse effect; however, developing low-cost electrocatalysts to mediate high-efficiency CO2 reduction remains a challenge on account of the limited understanding of the nature of the real active sites. Herein, we reveal the Znδ+ metalloid sites as the real active sites of stable nonstoichiometric ZnOx structure derived from Zn2 P2 O7 through operando X-ray absorption fine structure analysis in conjunction with evolutionary-algorithm-based global optimization. Furthermore, theoretical and experimental results demonstrated that Znδ+ metalloid active sites could facilitate the activation of CO2 and the hydrogenation of *CO2 , thus accelerating the CO2 -to-CO conversion. Our work establishes a critical fundamental understanding of the origin of the real active center in the zinc-based electrocatalysts for CO2 reduction reaction.

Bioactive diterpenes from Callicarpa longissima.

Investigation of the leaves and twigs of Callicarpa longissima resulted in the isolation of four new compounds (1-4), callilongisins A-D, and five known compounds, ursolic acid, 3-oxoanticopalic acid, (E)-6ß-hydroxylabda-8(17),13-dien-15-oic acid, 5-hydroxy-3,6,7,4'-tetramethoxyflavone, and artemetin. Compounds 1-3 are 3,4-seco-abietane-type diterpenoids, and compound 4 is an analogue of a labdenoic-type diterpene. The structure of compound 1 was confirmed by X-ray crystallographic analysis. Cytotoxicity against a human prostate cancer cell line (PC3) and anti-inflammatory activities of the isolated compounds were evaluated.

[Electrophysiology of two human hyperpolarization activated cyclic nucleotide-gated potassium channels expressed in HEK293 cells].

OBJECTIVE: To express the human HCN2 and HCN4 genes in HEK293 cells and investigate the electrophysiology of the expressed channel protein. METHODS: cDNA encoding human HCN2 or HCN4 gene was ligated into a shuttle vector pAdTrack-CMV. Homologous recombination was performed in Escherichia coli of the line BJ5183. Human embryonic kidney cells of the line 293 (HEK293 cells) were cultured and transfected with the positive recombinant adenovirus plasmid. Then the HEK293 cells were infected by AdhHCN2 or AdhHCN4 and the whole cell hyperpolarization-activated currents were recorded in HEK293 cells transfected with hHCN2 and hHCN4. RESULTS: If-like currents could be found in the HEK293 cells transfected with hHCN2 and hHCN4. The channels were activated by hyperpolarized potentials. Boltzmann equation showed that the half-activation voltage of the hHCN2 and hHCN4 channels were -114.8 mV +/- 3.3 mV and -125.9 mV +/- 2.9 mV respectively (P = 0.024). The reversal slope factors of the hHCN2 and hHCN4 channels were 11.1 mV +/- 1.2 mV and 13.7 mV +/- 1.3 mV respectively (P = 0.22). The activation kinetics was faster in hHCN2 than in hHCN4, with the activation constants at -110 mV being 0.99 s +/- 0.21 s and 8.47 s +/- 2.85 s respectively. The relative permeation ratio for sodium and potassium were 0.40 and 0.34 respectively in these two channels. Caesium chloride of the concentration of 2 mmol/L prominently inhibited both currents. CONCLUSION: The target genes hHCN2 and hHCN4 are successfully expressed in HEK293 cells, and the expressed functional channels have profoundly different activation kinetics.

[Electrophysiological effects of hyperpolarization activated cyclic nucleotide gated channel 4 overexpression in neonatal cardiomyocytes mediated by recombinant adenovirus].

OBJECTIVE: To explore the electrophysiological effects of hyperpolarization activated cyclic nucleotide gated channel 4 (HCN4) overexpression in rat neonatal cardiomyocytes mediated by recombinant adenovirus. METHODS: Ventricular cardiomyocytes were obtained from 20 neonatal rats. Recombinant adenovirus carrying HCN4 gene, AdHCN4, a dominant isoform of hyperpolarization activated cyclic nucleotide gated cation channel gene in cardiac transduction system, was constructed and used to transfect the neonatal rat ventricular cardiomyocytes. Untransfected cardiomyocytes were used as controls. RT-PCR and immunofluorescence cytochemistry were used to detect the HCN4 mRNA and protein expression. The electrophysiological characteristics of infected cardiomyocytes were studied by patch clamp. RESULTS: The mRNA and protein expression levels of HCN4 in AdHCN4 infected cardiomyocytes were both markedly higher than those of the control cardiomyocytes. AdHCN4 caused a significant increase in The spontaneous rate in the transfected cardiomyocytes was 92.5 + 7.4 bpm, significantly higher than that of the control cells (68.9 + 6.2 bpm, P < 0.05). Patch clamp experiments showed that the pacemaker current density in the AdHCN4 infected cardiomyocytes was 115.7 + 7.8 pA/pF, significantly higher than that of the untransfected cells (7.2 + 0.6 pA/pF, P < 0.05). CONCLUSION: Overexpression of HCN4 can enhance the autorhythmicity of neonatal cardiomyocytes and significantly increase the spontaneous beat rate. HCN4 channel gene may represent a candidate gene in gene therapy for bradycardia diseases.

[Transfecting rat heart with human pacemaker gene in vivo to create a biological pacemaker].

OBJECTIVE: To test the ability and safety of injecting the hHCN gene into the ventricle of intact rats to create a novel biological pacemaker and to explore the duration of the hHCN over-expression in vivo. METHODS: Adenoviral constructs incorporating hHCN4 and green fluorescent protein (GFP) were subepicardially injected into the rats' left ventricular wall in situ (n = 10). Control group was injected with adenoviral constructs of GFP alone (n = 10). ECGs were recorded every day within the initial three days after operation. Up to seven days after injection, all rats were anesthetized and subjected to cervical vagal trunks stimulation to permit the emergence of escape rhythm. Then pace-mapping was performed by the hand-hold electrode. Finally, hearts were removed for histology and immunofluorescence study. RESULTS: Ventricular arrhythmic events were not found during the first three days. During vagal stimulation, the rate of spontaneous rhythm in the hHCN4-injected group was significantly more rapid than that in the control group (69 bpm +/- 6 bpm vs 41 bpm +/- 10 bpm, P < 0.001). Pace-mapping confirmed the escape rhythm in the hHCN4-injected group was originated near the injection site. Immunofluorescence study showed the overexpression of hHCN4 on the site of injection. However, the duration of hHCN overexpression was no more than one month after injection. CONCLUSION: The overexpression of hHCN4 provides ventricular escape rhythm with the physiologically acceptable rate. Long-term feasibility of this approach needs to be tested.

Control design for signal transduction networks.

Signal transduction networks of biological systems are highly complex. How to mathematically describe a signal transduction network by systematic approaches to further develop an appropriate and effective control strategy is attractive to control engineers. In this paper, the synergism and saturation system (S-systems) representations are used to describe signal transduction networks and a control design idea is presented. For constructing mathematical models, a cascaded analysis model is first proposed. Dynamic analysis and controller design are simulated and verified.