• The Korean Journal of Physiology
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• v.25 no.1
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• pp.17-26
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• 1991

Single smooth muscle cells of the rabbit pulmonary artery were isolated by treatment with collagenase and elastase. Using the patch clamp technique, potassium channel activity was recorded from the inside-out membrane patch. The channel had a sin히e channel conductance of about 360 pS in symmetrical concentration of K on both sides of the patch, 150 mM, and had a linear current-voltage relationship. During the application of 10 mM tetraethylammonium (TEA) to the intracellular membrane surface, the amplitude of single channel current was reduced and very rapid flickering appeared. The open probability $(P_0)$ of this channel was increased by increasing positivity of the potential across the patch membrane, with e-fold increase by 20 mV depolarization, and by increasing the internal $Ca^{2+}$ concentration. These findings are consistent with those of large conductance Ca-activated K channels reported in other tissues. But the shortening of the mean open time by increasing $[Ca^{2+}]_i$, was an unexpected result and one additional closed state which might be arisen from a block of the open channel by Ca binding was suggested. The $P_0-membrane$ potential relationship was modulated by internal pH. Decreasing pH reduced $P_0$. Increasing pH not only increased $P_0$ but also weakened the voltage dependency of the channel opening. The modulation of Ca-activated K channel by pH was thought to be related to the mechanism of regulation of vascular tone by the pH change.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.5
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• pp.397-404
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• 2022

Fesoterodine, an antimuscarinic drug, is widely used to treat overactive bladder syndrome. However, there is little information about its effects on vascular K⁺ channels. In this study, voltage-dependent K⁺ (Kv) channel inhibition by fesoterodine was investigated using the patch-clamp technique in rabbit coronary artery. In whole-cell patches, the addition of fesoterodine to the bath inhibited the Kv currents in a concentration-dependent manner, with an IC₅₀ value of 3.19 ± 0.91 μM and a Hill coefficient of 0.56 ± 0.03. Although the drug did not alter the voltage-dependence of steady-state activation, it shifted the steady-state inactivation curve to a more negative potential, suggesting that fesoterodine affects the voltage-sensor of the Kv channel. Inhibition by fesoterodine was significantly enhanced by repetitive train pulses (1 or 2 Hz). Furthermore, it significantly increased the recovery time constant from inactivation, suggesting that the Kv channel inhibition by fesoterodine is use (state)-dependent. Its inhibitory effect disappeared by pretreatment with a Kv 1.5 inhibitor. However, pretreatment with Kv2.1 or Kv7 inhibitors did not affect the inhibitory effects on Kv channels. Based on these results, we conclude that fesoterodine inhibits vascular Kv channels (mainly the Kv1.5 subtype) in a concentration- and use (state)-dependent manner, independent of muscarinic receptor antagonism.

• BMB Reports
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• v.32 no.3
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• pp.273-278
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• 1999

Cytolysin produced by Vibrio vulnificus has been known to be lethal to mice by increasing vascular permeability and neutrophil sequestration in the lung. In the present study, a cytotoxic mechanism of V. vulnificus cytolysin on the neutrophil was investigated. Cytolysin rapidly bound to neutrophils and induced cell death, as determined by the trypan blue exclusion test. V. vulnificus cytolysin caused the depletion of cellular ATP without the release of ATP or lactate dehydrogenase. Formation of transmembrane pores was evidenced by the rapid efflux of potassium and 2-deoxy-D-[$^3H$]glucose from cytolysin-treated neutrophils. It was further confirmed by the rapid flow of monovalent ions in the patch clamp of cytolysin-treated neutrophil membrane. The pore formation was accompanied by the oligomerization of cytolysin monomers on the neutrophil membrane as demonstrated by immunoblot, which exhibited a 210 kDa band corresponding to a tetramer of the native cytolysin of $M_r$ 51,000. These findings indicate that V. vulnificus cytolysin rapidly binds to the neutrophil membrane and oligomerizes to form small transmembrane pores, which induce the efflux of potassium and the depletion of cellular ATP leading to cell death without cytolysis.

• Annals of Hepato-Biliary-Pancreatic Surgery
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• v.27 no.2
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• pp.131-140
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• 2023

During minimally invasive liver resection (MILR), the Pringle maneuver aims to minimize blood loss and provide a clear operative field, thereby identifying intrahepatic structures and facilitating safe parenchymal transection. Several techniques for using the Pringle maneuver in MILR have been described. This review presents various methods which have been reported in the literature. A systematic literature search used the MEDLINE/PubMed database from its earliest records to August 2022 using appropriate search headings and keywords. The primary outcome was identifying techniques for performing hepatic inflow occlusion during laparoscopic/robotic hepatectomy. Inclusion criteria consisted of publications describing technical steps to obtain hepatic inflow occlusion during minimally invasive hepatectomy. A literature search identified 23 relevant publications, and the full texts were examined. The techniques described in the reports can be broadly categorized into three groups: (1) the Rummel-tourniquet technique, (2) vascular clamp use, and (3) the Huang Loop technique. Various techniques have been used in MILR to achieve inflow confinement successfully. The authors prefer the modified Huang Loop technique because it is inexpensive, reliable, and quick to apply or release. Hepatobiliary surgeons are advised to familiarize themselves with these MILR techniques, which have proven effective and safe inflow occlusion.

• Archives of Plastic Surgery
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• v.37 no.5
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• pp.547-554
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• 2010

Purpose: The purpose of this study is to delineate the optimal time of venous revascularization for preventing the flap necrosis due to venous occlusion, and to clarify the usefulness of tissue oxygen pressure ($TcpO_2$) in the determination of the point of time for venous revascularization. Methods: Thirty-six, $3{\times}3\;cm$ sized epigastric island flap was elevated in left abdomen of male Sprague-Dawley rat weighing 250 gram. Flaps were randomly assigned to six groups of six flaps according to the duration of venous occlusion with microvascular clamp; 10 minutes in the group I as the control, 60 minutes in the group II, 2 hours in the group III, 3 hours in the group IV, 4 hours in the group V, and 6 hours in the group VI, respectively. Just before removal of clamp after flap was reposed in situ, the ratio of $TcpO_2$ (tissue oxygen pressure) of the island flap to that of right abdomen was calculated in each group, and tissue specimen was harvested from the distal area of the flap for histological evaluation of vascular change. Five days later, survival area of the flap was estimated, and evaluated the correlation between the tissue oxygen pressure and the rate of flap survival. Results: The $TcpO_2$ and the survival rate of flap were decreased proportionally with the duration of venous occlusion. The ratio of the $TcpO_2$ of the flap is decreased abruptly to below sixty percentile compared to the $TcpO_2$ of normal tissue, and the survived area of the flap is decreased to nine-tenth of the designed size after three hours of total venous occlusion. Histologically, the number of congested vessels was increased according to venous occluded time, and proportionally increased after 3-hours of occlusion significantly. Conclusion: There is a close correlation between the $TcpO_2$ and the survival rate of flaps according to the duration of venous occlusion. Therefore, the $TcpO_2$ represents the hemodynamic changes within the flap, and thought to be an alternative effective tool in the flap monitoring for venous revascularization.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.5
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• pp.284-293
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• 2013

Purpose: For reconstruction of craniomaxillofacial defects caused by tumor, trauma, infection etc, free flap transplantation with microvascular surgery is a very useful method. Thrombus formation at the anastomosis site is the major cause of graft failure. 4-Hexylresorcinol (4-HR) is generally known as an antiseptic and antiparasitic agent. This study was conducted in order to evaluate the effect of 4-HR on blood coagulation in vitro. In addition, we investigated thrombus formation and endothelial repair of an injured vessel in an animal model. Methods: In the in vitro experiment, we compared blood coagulation time between the 4-HR treated group and normal blood. Thirty rats were used for in vivo animal experiments. After exposure of the right femoral vein, a micro vessel clamp was placed and the femoral vein was intentionally cut. Microvascular anastomosis was performed on all rats using 10-0 nylon under microscopy. The animals were divided into two groups. In the experimental group (n=15), 4-HR (250 mg/kg) mixed with olive oil (10 mL/kg) was administered per os daily. Animals in the control group (n=15) were given olive oil only. The animals were sacrificed at three days, seven days, and fourteen days after surgery and rat femoral vein samples were taken. Vascular patency and thrombus formation were investigated just before sacrifice. Histologic analysis was performed under a microscope. Results: Results of an in vitro blood coagulation test showed that coagulation time was delayed in the 4-HR treated group. The results obtained from an in vivo 4-HR administered rat model showed that the patency of all experimental groups was better at thirty minutes, seven days, and fourteen days after microvascular anastomosis than that of the control group at seven and fourteen days after anastomosis, and the amount of thrombus in the experimental groups was much less than that of the control group. Endothelial repair was observed in the histologic analysis. Conclusion: Findings of this study demonstrated that blood coagulation was delayed in the vitro 4-HR treated group. In addition, good vascular patency, anti-thrombotic effect, and repair of venous endothelial cells were observed in the vivo 4-HR administered rat group.

• Korean Journal of Head & Neck Oncology
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• v.20 no.2
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• pp.177-180
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• 2004

Objectives: To completely excise a malignant tumor which invades carotid artery walls, the resection and reconstruction of the carotid artery is essentially required. In most of the cases, interposition graft using a saphenous vein has been performed, however the vessel caliber discrepancy between a common carotid artery and the saphenous vein can result in a problem on surgical technique. We have introduced and evaluated a new titanium hemoclip tailing method to overcome vessel caliber discrepancy in interposition saphenous vein graft for carotid artery resection in the treatment of head and neck cancers. Method: After carotid artery resection, the calibers of the proximal common carotid artery and the vein were compared, and an orifice of the common carotid artery was gradually reduced to a little larger than or the same size as the orifice of the vein by using the titanium hemostatic clip. Subsequently, the common carotid artery was connected to the vein through anastomosis. The same method was also applied to the distal anastomosis site. There after, the vessels were connected through the anastomosis, and a circulation was restored by releasing a vascular clamp. Then, a titanium hemostatic clip-applied redundant portion on the outside of carotid artery was sutured by the blanket edge suture method, using 6-0 Prolene. Results: We have experienced this method in two patients with recurrent squamous cell carcinoma and neuroblostoma, respectively. The interposition saphenous vein graft of these patients was found to maintain good patency on the follow up angiography after one year, and they had no specific vascular complication, such as atherosclerosis. Conclusion: This method made it possible to simply perform the interposition saphenous vein graft (ISVG) within a short time and, therefore, was very useful for shortening the duration to block circulation.

• Proceedings of the Korean Biophysical Society Conference
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• 1999.06a
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• pp.61-62
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• 1999

;The mechanisms inducing hypertension are actively investigated and are still challenging topics. Basically hypertension must be caused by the disorder of $Ca^{2+}$ metabolism in vascular smooth muscle, such as the increase of $Ca^{2+}$ influx, the decrease of ci+ efflux, or the change of sensitivity of contractile protein etc. The one of cause of the increase of ci+ influx may be the change of ci+ channel activity. Even though the relationships of ci+ channel activity and hypertension were studied using various hypertension models, still it is not clear how much change of $Ca^{2+}$ channel activity in diabetes mellitus (DM) induced hypertension is occurred. We induced DM hypertension in SD rat and compared the $Ca^{2+}$ channel activity with age-matched normotensive SD rat. For inducing DM hypertension, left kidney was removed with 200 gm rat and, after 1 month, 60 mg/kg of streptozotocin was injected into peritoneal space to induce diabetes mellitus. Usually after 4-6 weeks, hypertension was fully induced. For isolating vascular smooth muscle cells (VSMC), we used mesenteric arteriole (3rd - 4th branch of mesenteric artery) of which diameter is below 150 urn. VSMCs were isolated enzymatically. $Ca^{2+}$ current was measured using whole cell patch clamp technique. All experiments were performed at $37^{\circ}C$. The cell membrane area of VSMC of DM hypertensive rat is larger than that of control VSMC($36.6{\pm}3.64{\;}pF{\;}vs{\;}22.4{\pm}1.29{\;}pF, {\;}mean{\pm}S.E.$) When we compared the current amplitude, the $Ca^{2+}$ current amplitude in VSMC of DM hypertensive rat is much larger than that in VSMC of normotensive age-matched rat. After $Ca^{2+}$ current amplitude was normalized by cell membrane area, the current amplitude in DM hypertension is increased to $249.1{\pm}15.9{\;}%{\;}(mean{\pm}S.E.M)$, which means the ;absolute current amplitude is about 4 times larger in DM hypertension. When we compared the steady state activation and inactivation. there were no noticeable differences. From these results. one of cause of the DM hypertension is due to the increase of $Ca^{2+}$ current amplitude. But it need further study why the $Ca^{2+}$ current is so large in VSMC of DM hypertension and how much $Ca^{2+}$ influx through $Ca^{2+}$ channel contribute to the increase of intracellular $Ca^{2+}$ and eventually contribute to development of hypertension.ypertension.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.2
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• pp.169-183
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• 1997

We have investigated the two types of voltage-dependent outward potassium (K) currents, i.e. delayed rectifier K current ($I_{K(V)}$) and 'A-like' transient outward K current ($I_{to}$) with patch-clamp technique in single smooth muscle cells (SMCs) isolated from rabbit basilar artery, and investigated the characteristics of them. The time-courses of activation were well fitted by exponential function raised to second power ($n^2$) in $I_{K(V)}$ and fourth power ($n^4$) in $I_{to}$. The activation, inactivation and recovery time courses of $I_{to}$ were much faster than that of $I_{K(V)}$. The steady-state activation and inactivation of $I_{K(V)}$ was at the more hyperpolarized range than that of $I_{to}$ contrary to the reports in other vascular SMCs. Tetraethylammonium chloride (TEA; 10 mM) markedly inhibited $I_{K(V)}$ but little affected $I_{to}$. 4-Aminopyridine (4-AP) had similar inhibitory potency on both currents. While a low concentration of $Cd^{2+}$ (0.5 mM) shifted the current- voltage relationship of $I_{to}$ to the positive direction without change of maximum conductance, $Cd^{2+}$ did not cause any appreciable change for $I_{K(V)}$.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.5
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• pp.611-616
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• 1998

Although the $Ca^{2+}-activated\;K^+\;(I_{K,Ca})$ channel is known to play an important role in the maintenance of resting membrane potential, the regulation of the channel in physiological condition is not completely understood in vascular myocytes. In this study, we investigated the role of cytoplasmic $Mg^{2+}$ on the regulation of $I_{K,Ca}$ channel in pulmonary arterial myocytes of the rabbit using the inside-out patch clamp technique. $Mg^{2+}$ increased open probability (Po), but decreased the magnitude of single channel current. $Mg^{2+}-induced$ block of unitary current showed strong voltage dependence but increase of Po by $Mg^{2+}$ was not dependent on the membrane potential. The apparent effect of $Mg^{2+}$ might, thus, depend on the proportion between opposite effects on the Po and on the conductance of $I_{K,Ca}$ channel. In low concentration of cytoplasmic $Ca^{2+},\;Mg^{2+}$ increased $I_{K,Ca}$ by mainly enhancement of Po. However, at very high concentration of cytoplasmic $Ca^{2+},$ such as pCa 5.5, $Mg^{2+}$ decreased $I_{K,Ca}$ through the inhibition of unitary current. Moreover, $Mg^{2+}$ could activate the channel even in the absence of $Ca^{2+}.\;Mg^{2+}$ might, therefore, partly contribute to the opening of $I_{K,Ca}$ channel in resting membrane potential. This phenomenon might explain why $I_{K,Ca}$ contributes to the resting membrane potential where membrane potential and concentration of free $Ca^{2+}$ are very low.