Effects of Ca^(2+) channel blockers on store-operated Ca^(2+) channel currents of Kupffer cells after hepatic ischemia/reperfusion injury in rats

AIM： To study the effects of hepatic ischemia/ reperfusion （I/R） injury on store-operated calcium channel （SOC） currents （Isoc） in freshly isolated rat Kupffer cells, and the effects of Ca^2＋ channel blockers, 2-aminoethoxydiphenyl borate （2-APB）, SK＆F96365, econazole and miconazole, on Isoc in isolated rat Kupffer cells after hepatic I/R injury.METHODS： The model of rat hepatic I/R injury was established. Whole-cell patch-clamp techniques were performed to investigate the effects of 2-APB, SK＆F96365, econazole and miconazole on Isoc in isolated rat Kupffer cells after hepatic I/R injury.RESULTS： I/R injury significantly increased Isoc from -80.4±25.2pA to -159.5±34.5pA （^bp 〈 0.01, n = 30）. 2-APB （20, 40, 60, 80, 100 pmol/L）, SK＆F96365 （5, 10, 20, 40, 50 pmol/L）, econazole （0.1, 0.3, 1, 3, 10 μmol/L） and miconazole （0.1, 0.3, 1, 3, 10 μmol/L） inhibited Isoc in a concentration-dependent manner with IC50 of 37.41 μmol/L （n = 8）, 5.89 μmol/L （n = 11）, 0.21 μmol/L （n = 13）, and 0.28 μmol/L （n = 10）. The peak value of Isoc in the I-V relationship was decreased by the blockers in different concentrations, but the reverse potential of Isoc was not transformed. CONCLUSION： SOC is the main channel for the influx of Ca^2＋ during hepatic I/R injuries. Calcium channel blockers, 2-APB, SK＆F96365, econazole and miconazole,have obviously protective effects on I/R injury, probably by inhibiting Isoc in Kupffer cells and preventing the activation of Kupffer cells.

Macrophage migration inhibitory factor decreased T-type Ca^(2+) channel current through activating Src

Aims T-type Ca2+ current(ICaT)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 Ca2+ channel in atrium myocytes.Methods We used whole-cell voltage-clamp technique and biochemical assays to study the regulation and expression of ICa,T in mouse atrium myocytes（HL-1 cells）.Results Serum MIF concentrations was slightly increased in patients with AF compared to sinus rhythm（SR） controls.In cultured HL-1 cells, significant amounts of MIF were produced in response to hydrogen peroxide（H2O2）,but not AngiotensinⅡ（AngⅡ）. Mouse recombinant MIF（rMIF）（20 or 40 nM,24 h） suppressed peak ICa,T by-38%and-60%in a concentration-dependent manner,impaired the voltage-dependent activation of ICa,T,and down-regulated of TCC alG mRNA.Src inhibitors genistein and PPl significantly enhanced ICaT.The depression of ICa,T induced by rMIF could be reversed by genistein and PP1.Conclusions MIFis involved in the pathogenesis of AF,probably by decreasing ICa,T through impairment of the channel function and activation of c-Src kinases in atrium myocytes.

Effect of gingerol on colonic motility via inhibition of calcium channel currents in rats

AIM: To investigate the effect of gingerol on colonic motility and the action of L-type calcium channel currents in this process.METHODS: The distal colon was cut along the mesenteric border and cleaned with Ca^(2+)-free physiological saline solution. Muscle strips were removed and placed in Ca^(2+)-free physiological saline solution, which was oxygenated continuously. Longitudinal smooth muscle samples were prepared by cutting along the muscle strips and were then placed in a chamber. Mechanical contractile activities of isolated colonic segments in rats were recorded by a 4-channel physiograph. Colon smooth muscle cells were dissociated by enzymatic digestion. L-type calcium currents were recorded using the conventional whole-cell patch-clamp technique.RESULTS: Gingerol inhibited the spontaneous contraction of colonic longitudinal smooth muscle in a dose-dependent manner with inhibition percentages of 13.3% ± 4.1%, 43.4% ± 3.9%, 78.2% ± 3.6% and 80.5% ± 4.5% at 25 μmol/L, 50 μmol/L, 75 μmol/L and 100 μmol/L, respectively(P < 0.01). Nifedipine, an L-type calcium channel blocker, diminished the inhibition of colonic motility by gingerol. Gingerol inhibited L-type calcium channel currents in colonic longitudinal myocytes of rats. At a 75 μmol/L concentration of gingerol, the percentage of gingerolinduced inhibition was diminished by nifedipine from 77.1% ± 4.2% to 42.6% ± 3.6%(P < 0.01). Gingerol suppressed IBa in a dose-dependent manner, and the inhibition rates were 22.7% ± 2.38%, 35.77% ± 3.14%, 49.78% ± 3.48% and 53.78% ± 4.16% of control at 0 m V, respectively, at concentrations of 25 μmol/L, 50 μmol/L, 75 μmol/L and 100 μmol/L(P < 0.01). The steady-state activation curve was shifted to the right by treatment with gingerol. The value of half activation was-14.23 ± 1.12 m V in the control group and-10.56 ± 1.04 m V in the 75 μmol/L group(P < 0.05) with slope factors, Ks, of 7.16 ± 0.84 and 7.02 ± 0.93(P < 0.05) in the control and 75 μmol/L groups, respectively. However, the steady-state inactivation curve was not changed, with a half-inactivation voltage, 0.5 V, of-27.43 ± 1.26 m V in the control group and-26.56 ± 1.53 m V in the 75 μmol/L gingerol group(P > 0.05), and a slope factor, K, of 13.24 ± 1.62 in the control group and 13.45 ± 1.68(P > 0.05) in the 75 μmol/L gingerol group.CONCLUSION: Gingerol inhibits colonic motility by preventing Ca^(2+) influx through L-type calcium channels.

Effects of 2-APB on Store-operated Ca^(2+) Channel Currents of Hepatocytes after Hepatic Ischemia/Reperfusion Injury in Rats

The effects of hepatic ischemia/reperfusion (I/R) injuries on hepatocellular viability and store-operated calcium current (Isoc) in isolated rat hepatocytes and the effects of 2-APB on store-operated calcium current (Isoc) in isolated rat hepatocytes after hepatic ischemia/reperfusion injuries were studied. Hepatic ischemia and reperfusion injury model was established and whole cell patch-clamp techniques were used to investigate the effects of 2-APB on Isoc. The results showed that ischemia/reperfusion injuries could significantly reduce hepatocellular viability and further increase Isoc in hepatocytes and 2-APB (20, 40, 60, 80, 100 μmol/L) produced a concentration-dependent decrease of Isoc with IC 50 value of 64.63±10.56 μmol/L (n=8). It was concluded that ischemia/reperfusion injuries could reduce hepatocellular viability, probably through increased Isoc in hepatocytes and 2-APB had a protective effect on ischemia/reperfusion-induced liver injury, probably though inhibiting Isoc.

Connexin 43 co-locolizes and regulates the L type calcium channel current in atrial myocytes

Background Atrial fibrillation （AF） is the most common sustained cardiac arrhythmia without effective treatment. AF is associated with atrial conduction disturbances caused by electrical and / or structural remodeling. But the role of connexin （Cx） 43 in the regulation of L type calcium channel （LCC） remains unclear. We hypothesized that Cx 43 might co-localize and regulate the L type calcium channel current （ICa, D. Methods Real-time PCR and whole-cell patch clamp were used to detect the expression of LCC lc subunit and the cur rent density of Ica, L, before and after Cx 43 knocking down respectively. The co-localization of Cx 43 with LCC was investigated by co-immunoprecipitation and confocal microscopy. Results Knocking down of Cx43 significantly inhibited the current density of ICa, L through decreasing the gene expression of LCC alc in cul tured atrium-derived myocytes （HL-1 cells）. Cx43 co-localized with LCC eric subunit in atrial myocytes. Conclusions Cx 43 regulates the ICa, L in atrial myoctyes through LCC, representing a potential pathogenic mechanism in atrial arrhythmias.

EFFECT OF COXSACKIEVIRUS B3 ON ION CHANNEL CURRENTS IN RAT VENTRICULAR MYOCYTES

To investigate the effects of coxsackievirus B 3(CVB 3) on ion channel currents in rat ventricular myocytes. Methods.Rat hearts were isolated with collagenase to acquire single ventricular myocytes, L type voltage dependent calcium channel(VDCC)current (I Ca ),Na + current (I Na ), outward potassium current (I out ), inwardly rectifying potassium current(I KI ) were recorded using whole cell patch clamp techniques. ResultsCVB 3 infection increased I Ca and I out , while decreased I KI ; but it had no obvious effect on I Na . Conclusion.The effects of CVB 3 on I Ca 、 I out 、 I KI may be one of the mechanisms of myocytes damage and the occurrence of abnormal electroactivities induced by CVB 3 infection.

High-Performance In0.23Ga0.77As Channel MOSFETs with High Current Ratio Ion/Ioff Grown on Semi-insulating GaAs Substrates by MOCVD

We demonstrate high-performance In0.23 Ga0.77 As channel metal-oxide-semiconductor field-effect transistors （ MOS- FETs） with high on-current to off-current （Ion/Ioff） ratio grown on semi-insulating GaAs wafers by metal-organic chemical vapor deposition （MOCVD）. The 2μm channel-length devices exhibit a peak extrinsic transeonductance of 150 mS/mm and a drain current up to 500 mA/mm. The maximum effective mobility is 1680 cm2/Vs extracted by the split C-V method. Furthermore, the Ion/Ioff ratio is significantly improved from approximately 4.5 × 10^3 up to approximately 4.32 × 10^4 by controlling the etch thickness of In0.49Ga0.51P, The high drain current and high Ion/Ioff ratio of the In0.23Ga0.77As channel MOSFETs are achieved due to the high effective mobility and the low gate leakage current density.

Reversal current observed in micro-and submicro-channel flow under non-continuous DC electric field

In practical applications of biochips and bio-sensors, electrokinetic mechanisms are commonly employed to manipulate and analyze the characteristics of single bio-molecules. To accurately and flexibly control the movement of single molecule within micro-/submicro-fluidic channels, the characteristics of current signals at the initial stage of the flow are systematically studied based on a three-electrode system. The current response of micro-/submicro-fluidic channels filled with different electrolyte solutions in non-continuous external electric field are investigated. It is found, there always exists a current reversal phenomenon, which is an inherent property of the current signals in micro/submicro-fluidics Each solution has an individual critical voltage under which the steady current value is equal to zero The interaction between the steady current and external applied voltage follows an exponential function. All these results can be attributed to the overpotentials of the electric double layer on the electrodes. These results are helpful for the design and fabrication of functional micro/nano-scale fluidic sensors and biochips.

Anomalous Channel Length Dependence of Hot-Carrier-Induced Saturation Drain Current Degradation in n-Type MOSFETs

The dependencies of hot-carrier-induced degradations on the effective channel length Lch,eff are investigated for n-type metal-oxide-semiconductor field effect transistor （MOSFETs）. Our experiments find that, with decreasing Lch,eff, the saturation drain current （Iasat ） degradation is unexpectedly alleviated. The further study demonstrates that the anomalous Lch,eff dependence of Idsat degradation is induced by the increasing influence of the substrate current degradation on the lazar degradation with Lch,eff reducing.

Effects of Intrathecally Administerd NaV1.8 Antisense Oligonucleotide on the Expression of Sodium Channel mRNA in Dorsal Root Ganglion

Neuropathic pain has been hypothesized to be the result of aberrant expression and function of sodium channels at the site of injury. To investigate the effects of NaV1.8 antisense oligonucleotide on the expression of sodium channel mRNA in dorsal root ganglion （DRG） neurons in chronic neuropathic pain. 24 Sprague-Dawley rats weighing 200--260 g were anesthetized with the intraperitoneal injection of 300 mg· kg^-1 choral hydrate. The CCI model was made by loose ligation of sciatic nerve trunk by 4--0 chromic gut. The mechanical and thermal pain threshold were measured before operation and 1, 3, 5, 7, 9, 11, 13 days after operation. A PE-10 catheter was implanted in subarachnoid space at lumbar region. On the 7th postoperative day the animals were randomly divided into 4 groups. The drugs were injected intrathecally twice a day for 5 consecutive days in group 2--4. The animals were decapitated 14 days after the surgery. The L4--L6 DRG of the operated side was removed and crushed, and total RNA was extracted with Trizol reagent. The contralateral side was used as control. The change of NaV1.8 sodium channel transcripts was determined by RT-PCR. Pain threshold was significantly lowered after CCI as compared with that in control group and was elevated 3 days after antisense oligonucleotide injection. Sensory neuron specific TTX-R sodium channel NaV1.8 transcript was down-regulated after antisense oligonucleotide injection at the dosage of 45 μg as compared with that in CCI group （P〈0.01）, and it was even greater at the dosage of 90 μg. The intrathecally injected NaV1.8 antisense oligonucleotide can reduce the mechanical allodynia and thermal hyperalgesia partially by downregulating the SNS transcript expression.

Voltage- and Use-dependent Effect of 7-chlor-benzylte-trahydropalmatine on Sodium Currents in Guinea Pig Ventricular Myocytes

The whole-cell patch-clamp technique was employed to obtain information about the voltage-dependence and kinetics of interaction of 7-chlor-benzylte-trahydropalmatine (7-Cl-BTHP) with cardiac sodium channels. 7-Cl-BTHP (30 mol/L) significantly decreased the peak sodium current (from 7. 8±1. 8 nA to 5. 3±1. 4 nA, P<0. 01, n=5), without producing a shift of the current-voltage curve. It shifted the inactivation curves of sodium current to hyperpolarized potentials, and the V(0.5) was shifted from - (82. 5±2. 5) mV to - (95±2.4) mV (P <0. 05, n=4). 7-Cl-BTHP produced a significant use-dependent effect that was proportional to the duration of the voltage step. In addition, 7-Cl-BTHP slowed the recovery of sodium channel from inactivation, which could explain its use-dependent effects on sodium current. The characteristics of 7-Cl-BTHP blockage suggest that this agent binds preferentially to inactivated sodium channels.

Characteristics of Transient Outward Potassium Channel Exposed to 3 mT Static Magnetic Field

Acutely isolated mouse hippocampal CA3 pyramidal neurons were exposed to 3 mT static magnetic field,and the characteristics of transient outward K+ channel were studied using the whole-cell patch-clamp technique.The experiment revealed that the amplitude of transient outward potassium channel current was reduced.The maximum activated current densities of control group and exposure group were 163.62±20.68 pA/pF and 98.74±16.57 pA/pF(n=12,P<0.01) respectively.The static magnetic field exposure affected the activation and inactivation process of transient outward potassium channel current.Due to the magnetic field exposure,the half-activation voltage of the activation curves changed from 5.59±1.96 mV to 27.87±7.24 mV(n=12,P<0.05) ,and the slope factor changed from 19.43±2.11 mV to 25.87±4.22 mV(n=12,P<0.05) .The half-inactivation voltage of the inactivation curves also changed from-56.09±0.89 mV to-57.16±1.10 mV(n=12,P>0.05) and the slope factor of the inactivation curves from 8.69±0.80 mV to 10.87±1.02 mV(n=12,P<0.05) .The results show that the static magnetic field can change the characteristics of transient outward K+ channel,and affect the physiological functions of neurons.

The Theory of Field-Effect Transistors:XI. The Bipolar Electrochemical Currents(1-2-MOS-Gates on Thin-Thick Pure-Impure Base)

The field-effect transistor is inherently bipolar, having simultaneously electron and hole surface and volume channels and currents. The channels and currents are controlled by one or more externally applied transverse electric fields. It has been known as the unipolar field-effect transistor for 55-years since Shockley＇s 1952 invention,because the electron-current theory inevitably neglected the hole current from over-specified internal and boundary conditions, such as the electrical neutrality and the constant hole-electrochemical-potential, resulting in erroneous solutions of the internal and terminal electrical characteristics from the electron channel current alone, which are in gross error when the neglected hole current becomes comparable to the electron current, both in subthreshold and strong inversion. This report presents the general theory, that includes both electron and hole channels and currents. The rectangular （ x, y, z） parallelepiped transistors,uniform in the width direction （z-axis）,with one or two MOS gates on thin and thick,and pure and impure base, are used to illustrate the two-dimensional effects and the correct internal and boundary conditions for the electric and the electron and hole electrochemical potentials. Complete analytical equations of the DC current-voltage characteristics of four common MOS transistor structures are derived without over-specification： the 1-gate on semi-infinite-thick impure-base （the traditional bulk transistor）, the 1-gate on thin impure-silicon layer over oxide-insulated silicon bulk （SOI） ,the 1-gate on thin impure-silicon layer deposited on insulating glass （SOI TFT）, and the 2-gates on thin pure-base （FinFETs）.

The Effect of Levobunolol Hydrochloride on the Calcium andPotassium Channels in Isolated Ventricular Myocytes of Guinea Pig

The effects of levobunolol hydrochlorid (Bun) on the type L calciumchannel currents (Ica) and delayed rectifier potassium channel currents (Ik) in isolated ventricular myocytes of guinea pig were studied by using patch clamp wholecell recording techniques. The results were showed that: 1) Bun caused a dosedependent decrease in Ica and a dose-dependent increase in Ik of the ventricular myocytes.The threshold concentrations of Bun for Ica and Ik were 10-8 mol/L and10-7 mol/L respectively. The maximum effective concentration of Bun for both Ica and Ik was 3 × 10-5 mol/L, and half-maximal concentration was 3 × 10-6 mol/L;2 ) Ik was blocked by 2× 10-6mol/L tetraethylammonium (TEA). A concentration of 3 × 10-6 mol/L Bun showed a decreasing effect on the Ica as revealed by the current-voltage relationship curve, i. e., Bun caused an elevation of the curve; 3)When Ica was blocked by 2 × 10-6 mol/L Isoptin (Verapamil), at a concentrationof 3 × 106- mol/L Bun showed an increasing effect on Ik and the effect could be blocked by TEA. The above-mentioned results indicated that Bun had an inhibito-ry effect on Ica and a fascilitatory effect on Ik The results suggested that themolecular mechanisms of antihypertensive, heart rate slowing and β-receptorblocking effects of Bun might be due to decrease of Ica and increase of Ik.

Variations of temperature, salinity and current in the southern tidal passage of the Hangzhou Bay, China

Field surveys covering a spring-neap tidal period were conducted to investigate the characteristics of tidal dynamics within a curved channel in the southern Hangzhou Bay, China. The channel has a maximum depth of more than 100 m with an average tidal range of 2.5 m, serving as the main tidal passage in the southern part of the Hangzhou Bay. Water salinity, temperature and velocity data were collected from the ship-based transects and mooring measurements. During flood tide, the tidal current intrudes into the Hangzhou Bay through the northern side of the channel with a maximum velocity of about 2 m/s, while retreats through the southern side during ebb tide with a maximum velocity of 1.8 m/s. Due to the pressure, density gradients, the Coriolis force and centrifugal effect, a lateral exchange flow is generated as the tidal current relaxes from flood to ebb. Salinity and temperature data show that the water in the channel is weakly stratified during both spring and neap tides in summer time.However, mixing in the middle region will be enhanced by the lateral circulation. Mooring data indicate that the temperature and salinity are varying at a frequency similar to tidal current but higher than sea level oscillation.Our results support the hypothesis that the high frequency salinity and temperature variations could be generated by combination of the tidal current and the lateral exchanging flow.

Two-dimensional analytical solution for compound channel flows with vegetated floodplains

This paper presents a two-dimensional analytical solution for compound channel flows with vegetated floodplains. The depth-integrated N-S equation is used for analyzing the steady uniform flow. The effects of the vegetation are considered as the drag force item. The secondary currents are also taken into account in the governing equations, and the preliminary estimation of the secondary current intensity coefficient K is discussed. The predicted results for the straight channels and the apex cross-section of meandering channels agree well with experimental data, which shows that the analytical model presented here can be applied to predict the flow in compound channels with vegetated floodplains.

Differential Effects of d, l-Sotalol and d-Sotalol on Isoproterenol-Increased Delayed Rectifier Outward Potassium Current in Guinea Pig Single Ventricular Myocytes

The aim of this study was to compare the effects of d, l-Sotalol and dSotalol on the delayed rectifier K+ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier K+ outward currents were measured in isolated guinea pig single myocytes using the whole-cell configuration of the patch-clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of -40 mV in three experimental protocols [control, isoproterenol (10^(9)mol/L - 10^(-6) mol/L ), and isoproterenol (10^(-9)mol/L - 10^(-6)mol/L ) plus either d, l-Sotalol (10^(-4) mol/L) or d-Sotalol (10^(-4) mol/L)]. IK tail currents were measured upon repolarization to -40 mV. It was found that Ik was significantly amplified in the presence. of isoproterenol (10^(-9) mol/L- 10^(-6) mol/L) plus d-Sotalol. At 10-8 mol/L isoproterenol, Ik was increased by 92. 7%±17. 1 % (P<0. 05) and 54. 3 %±13. 4 % after d-Sotalol addition (P<0. 05). In contrast, d, l-Sotalol completely conteracted the increase of iK by isoproterenol (<10^(-8) mol/L), and compared to control, Ic was decreased by 35. 6 % ±8. 1% at 10^(-8) mol/L isoproterenol plus d, l-Sotalol (P<0. 05). It is concluded that the β-adrenergic blocking property of d, l-Sotalol but not that of dSotalol maintains the delayed rectifier K+ outward current blockade in the presence of isoproterenol in guinea pig myocytes. This might contribute to a superior antiarrhythmic efficacy as compared to d-Sotalol.

ISFET Based DNA Sensor: Current-Voltage Characteristic and Sensitivity to DNA Molecules

Dependency of both source-drain current and current sensitivity of nanosize ISFET biosensor vs. concentration of DNA molecules in aqueous solution theoretically is investigated. In calculations it is carried out effects concerning charge carriers distribution in current channel and concerning carriers’ mobility behavior in high electrical fields in the channel. The influence of DNA molecules on the work of ISFET biosensors is manifested by a change in the magnitude of the gate surface charge. Starting with fairly low concentrations of DNA, ISFET sensors respond to the presence of DNA molecules in an aqueous solution which is manifested by modulation of channel conductance and therefore the source-drain current changes of the field-effect transistor. It is shown that the current sensitivity with respect to concentration of DNA molecules linearly depends on the source-drain voltage and reaches high values.

Analytical solutions for transverse distributions of stream-wise velocity in turbulent flow in rectangular channel with partial vegetation

The theory of poroelasticity is introduced to study the hydraulic properties of the steady uniform turbulent flow in a partially vegetated rectangular channel. Plants are assumed as immovable media. The resistance caused by vegetation is expressed by the theory of poroelasticity. Considering the influence of a secondary flow, the momentum equation can be simplified. The momentum equation is nondimensionalized to obtain a smooth solution for the lateral distribution of the longitudinal velocity. To verify the model, an acoustic Doppler velocimeter （ADV） is used to measure the velocity field in a rectangular open channel partially with emergent artificial rigid vegetation. Comparisons between the measured data and the computed results show that the method can predict the transverse distributions of stream-wise velocities in turbulent flows in a rectangular channel with partial vegetation.

Behavior of near-field dilution of thermal buoyant jet discharged horizontally in compound open-channel

The RNG κ-ε model considering the buoyancy effect, which is solved by the hybrid finite analytic method, is used to simulate the mixture of the horizontal round thermal buoyant jet in compound open channel flow. The mixing features near the spout and flowing characteristic of the secondary currents are studied by numerical simulation. Meanwhile, （1） the distribution of the measured isovels for stream-wise velocity, （2） secondary currents, （3） the distribution of the measured isovels for temperature of typical cross-section near the spout, were obtained by the three-dimensional Micro ADV and the temperature measuring device. Compared with experimental data, the RNG κ-ε model based on buoyancy effect can preferably simulate the jet which performs the bifurcation phenomenon, jet reattachment （Conada effect） and beach secondary currents phenomenon with the effect of ambient flow, buoyancy, and secondary currents of compound section and so on.