We thank the patients for participating in this study. We are also grateful to the Biomedicine Electron Microscopy Laboratory of Basic Medical Science School of Central South University, particularly to Xiaoying Wu and Jin Li for technical support. This work was supported by the National Key Research and Development Program (No. 2018YFC1311204).

The Institutional Review Board approved the study protocol, and samples of the dilated left atrium (LA) tissues in the dextrocardia cases were collected for electron microscopy after written informed consent had been obtained.
1. Surgical procedure and ablation
<ol>
	<li>Perform a median sternotomy and suspend the left lateral pericardium after opening.
	<ol>
		<li>Cannulate the ascending aorta and the superior and inferior vena cava (SVC, IVC) to establish mild hypothermic ...

<ol>
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A., Guler, A., Kaya, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitral+valve+replacement+in+a+patient+with+situs+inversus+and+dextrocardia.">Mitral valve replacement in a patient with situs inversus and dextrocardia.</a> The Thoracic and Cardiovascular Surgeon. 59 (5), 305-306 (2011).</li><li>Uchimuro, T., Fukui, T., Matsuyama, S., Tabata, M., Takanashi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitral+valve+replacement+in+dextrocardia+and+situs+inversus.">Mitral valve replacement in dextrocardia and situs inversus.</a> Kyobu Geka. 65 (10), 858-861 (2012).</li><li>Atsumi, Y., Tokunaga, S., Yasuda, S., Fushimi, K., Masuda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitral+valve+surgery+in+a+patient+with+dextrocardia+and+180+degrees+counter-clockwise+rotated+heart+due+to+congenital+agenesis+of+the+right+lung.">Mitral valve surgery in a patient with dextrocardia and 180 degrees counter-clockwise rotated heart due to congenital agenesis of the right lung.</a> Journal of Cardiac Surgery. 28 (6), 635-637 (2013).</li><li>Kikon, M., Kazmi, A., Gupta, A., Grover, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left-sided+approach+for+mitral+valve+replacement+in+a+case+of+dextrocardia+with+situs+solitus.">Left-sided approach for mitral valve replacement in a case of dextrocardia with situs solitus.</a> Interactive Cardiovascular and Thoracic Surgery. 17 (5), 900-902 (2013).</li><li>Khan, J. 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M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cox+maze+III+procedure+for+atrial+fibrillation:+long-term+efficacy+in+patients+undergoing+lone+versus+concomitant+procedures.">The Cox maze III procedure for atrial fibrillation: long-term efficacy in patients undergoing lone versus concomitant procedures.</a> The Journal of Thoracic and Cardiovascular Surgery. 126 (6), 1822-1827 (2003).</li><li>Gaynor, S. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prediction+of+sinus+rhythm+in+patients+undergoing+concomitant+Cox+maze+procedure+through+a+median+sternotomy.">Prediction of sinus rhythm in patients undergoing concomitant Cox maze procedure through a median sternotomy.</a> The Journal of Thoracic and Cardiovascular Surgery. 148 (3), 881-886 (2014).</li><li>Damiano, R. J., Voeller, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biatrial+lesion+sets.">Biatrial lesion sets.</a> Journal of Interventional Cardiac Electrophysiology. 20 (3), 95-99 (2007).</li><li>Lawrance, C. P., Henn, M. C., Damiano, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+ablation+for+atrial+fibrillation:+techniques,+indications,+and+results.">Surgical ablation for atrial fibrillation: techniques, indications, and results.</a> Current Opinion in Cardiology. 30 (1), 58-64 (2015).</li><li>Gammie, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multi-institutional+experience+with+the+CryoMaze+procedure.">A multi-institutional experience with the CryoMaze procedure.</a> The Annals of Thoracic Surgery. 80 (3), 876-880 (2005).</li><li>Gaita, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linear+cryoablation+of+the+left+atrium+versus+pulmonary+vein+cryoisolation+in+patients+with+permanent+atrial+fibrillation+and+valvular+heart+disease:+correlation+of+electroanatomic+mapping+and+long-term+clinical+results.">Linear cryoablation of the left atrium versus pulmonary vein cryoisolation in patients with permanent atrial fibrillation and valvular heart disease: correlation of electroanatomic mapping and long-term clinical results.</a> Circulation. 111 (2), 136-142 (2005).</li><li>Ad, N., Henry, L., Hunt, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+concomitant+cryosurgical+Cox-Maze+procedure+using+Argon+based+cryoprobes:+12+month+results.">The concomitant cryosurgical Cox-Maze procedure using Argon based cryoprobes: 12 month results.</a> The Journal of Cardiovascular Surgery. 52 (4), 593 (2011).</li><li>Ad, N., Holmes, S. 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Dextrocardia is a group of rare heart defects in which the heart is situated on the right side of the thoracic cavity instead of on the left side. One-third of all dextrocardia cases are mirror-image cases, which means the orientation of the cardiac chambers is a mirror image to levocardia (normally situated heart)12. It is estimated that situs inversus dextrocardia associated with situs inversus totalis occurs with an incidence of much less than 1 in 10,000-50,000 births1<...

Dextrocardia is a rare, congenital cardiac malformation in which the axis of the heart is indexed to the right side of the thoracic cavity. Dextrocardia with situs inversus totalis refers to all visceral organs, including the heart, being mirrored and is extremely rare1,2. Atrial fibrillation (AF) is the most common arrhythmia that affects millions of people and causes substantial morbidity and mortality, especially with respect to the increased risk of stroke3.
A literature survey was unable to demonstrate a definitive approach for the concomitant Cox-Maze procedure (CMP) and valve surgery in dextrocardia with situs inversus. There are only a few reports of mitral valve replacement in cases of dextrocardia with situs solitus and even less in situs inversus4,5,6,7,8,9. However, those strategies are not applicable to tricuspid valve surgery, not to mention the complex CMP-IV in dextrocardia.
This paper reports the surgical techniques and experience in three cases concerning the biatrial lesion set of the CMP-IV operation using cryoablation or radiofrequency ablation concomitantly with valve surgery in situs inversus dextrocardia. All surgeries succeeded in the maintenance of sinus rhythm (SR) at 12 months follow-up to each patient and at 48 months follow-up to the previous two. Electron microscopy was used to investigate the transmurality of the atrial cryoablation.
CASE PRESENTATION:
Patients and preoperative examination 
From February 2016 to September 2020, a 48-year-old female patient, a 55-year-old male patient, and a 39-year-old male patient were successively admitted to the cardiac center with similar complaints of palpitation, dyspnea, and easy fatigability on exertion lasting for years (Table 1). They all denied any familiar history of situs inversus dextrocardia or other cardiac health comorbidities. All patients were referred to electrocardiography (ECG), chest roentgenography (Figure 1), computed tomography (CT), and Doppler transthoracic echocardiography (TTE) routinely for preoperative examination.
Case 1: A 48-year-old woman presented with complaints of palpitation, dyspnea, and easy fatigability on exertion lasting for years. The anamnesis was unremarkable. During the physical examination, a grade 3 systolic blowing murmur was heard in the fifth intercostal space outside the midline of the right clavicle. X-ray plain film and CT scan showed that the contour of the dextrocardia was enlarged and situs inversus totalis. Transesophageal echocardiography revealed moderate to severe mitral regurgitation and mild tricuspid regurgitation after moderate mitral valve prolapse and tricuspid annular dilation. There was no thrombosis, and the diameter of the left atrium was 5.3 cm. AF with a rapid ventricular rate was detected by ECG.
Case 2: A 55-year-old man presented with similar symptoms for years. There was no history of cardiac health comorbidities; however, he had had a stroke about six months ago. On physical examination, a diastolic blowing murmur was heard at the left second intercostal space, radiating to the neck. Plain X-ray and CT scan revealed dextrocardia with an enlarged contour and situs inversus totalis. Transesophageal echocardiography showed moderate aortic valve regurgitation and mild to moderate tricuspid regurgitation following mild aortic prolapse and tricuspid annular dilation. The diameter of the left atrium was 4.5 cm without atrial thrombosis. AF with a rapid ventricular rate was detected by ECG.
Case 3: A 39-year-old male presented with progressive exertional dyspnea and intermittent palpitation without any history of situs inversus dextrocardia or other cardiac health complications. During the physical examination, a grade 3 systolic blowing murmur was heard in the fifth intercostal space outside the midline of the right clavicle. X-ray plain film and CT scan revealed that the contour of dextrocardia was enlarged and situs inversus totalis. Transesophageal echocardiography showed severe mitral valve regurgitation and tricuspid regurgitation after annular enlargement. There was no atrial thrombosis, and the left atrium diameter was 5.8 cm. A 24 h ambulatory ECG demonstrated paroxysmal AF with a total load of 165 min.
Diagnosis, assessment, and plan 
Case 1: A biatrial cryo-Cox-Maze IV procedure (cryo-CMP-IV) with mechanic mitral valve replacement and tricuspid annuloplasty were simultaneously performed.
Case 2: A biatrial cryo-CMP-IV procedure with concomitant mechanic aortic valve (AV) replacement and tricuspid annuloplasty was performed.
Case 3: A biatrial CMP-IV procedure, mitral valvuloplasty, and tricuspid annuloplasty were performed simultaneously.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CryoICE</td>
			<td>AtriCure, Cincinnati, Ohio</td>
			<td>CRYO2</td>
			<td>Cryoablation in case 1 and case 2 was performed exclusively using it with a flexible 10cm in length metal cryoprobe.</td>
		</tr>
		<tr>
			<td>Medtronic Open Pivot Standard mechanical MV</td>
			<td>Medtronic, Minneapolis, Minn</td>
			<td>709291 / MHV 500DM27 STD MITRAL</td>
			<td>A 27-mm MV was adopted in case 1.</td>
		</tr>
		<tr>
			<td>Medtronic Open Pivot Standard mechanical AV</td>
			<td>Medtronic, Minneapolis, Minn</td>
			<td>646871 / MHV 500FA23 STD AORTIC</td>
			<td>A 23-mm AV was adopted in case 2.</td>
		</tr>
		<tr>
			<td>bipolar radiofrequency pen</td>
			<td>AtriCure Inc., Cincinnati, OH</td>
			<td></td>
			<td>Maze-IV in case 3</td>
		</tr>
		<tr>
			<td>bipolar radiofrequency clamps</td>
			<td>AtriCure Inc., Cincinnati, OH</td>
			<td></td>
			<td>Maze-IV in case 3</td>
		</tr>
		<tr>
			<td>Goretex</td>
			<td>W.L. Gore &#38; Associates, Inc., Elkton, Maryland</td>
			<td></td>
			<td>A surgical suture made of polytetrafluoroethylene.</td>
		</tr>
		<tr>
			<td>rigid mitral ring</td>
			<td>Kingstron Bio, Suzhou, China</td>
			<td>Element Force ARM32</td>
			<td>A 32 mm ring was adopted in case 3.</td>
		</tr>
		<tr>
			<td>Tricuspid Sovering Band</td>
			<td>Sorin Group Italia S.r.l., VC, Italy</td>
			<td>SBG0730 / SB30T</td>
			<td>A 30 mm ring was adopted in case 1.</td>
		</tr>
		<tr>
			<td>Tricuspid Sovering Band</td>
			<td>Sorin Group Italia S.r.l., VC, Italy</td>
			<td>SQB0240 / SB30T</td>
			<td>A 30 mm ring was adopted in case 2.</td>
		</tr>
		<tr>
			<td>Tricuspid Sovering Band</td>
			<td>Sorin Group Italia S.r.l., VC, Italy</td>
			<td>SBF0930 / SB28T</td>
			<td>A 28 mm band was adopted in case 3.</td>
		</tr>
	</tbody>
</table>

cox-maze iv procedure concomitant with valvular surgery in situs inversus dextrocardia: a single-center experience in china

Atrial fibrillation (AF) is the most common cardiac arrhythmia. The use of ablation technologies made the Cox-Maze IV procedure (CMP-IV) technically easier, faster, becoming the gold standard for the surgical treatment of AF. However, the efficacy and safety of CMP-IV in situs inversus dextrocardia are largely unknown. This paper summarizes the CMP-IV procedure performed concomitantly with valvular surgery in patients with situs inversus dextrocardia at this institution.
From February 2016 to September 2020, three dextrocardia patients with persistent AF and valvular diseases were referred to this institution for valvular and CMP-IV surgery. CMP-IV was performed using either cryoablation with a nitrous oxide (N2O)-based cryoprobe or a bipolar radiofrequency clamp and bipolar radiofrequency pen. Mechanical valve replacement or mitral vavuloplasty was performed in another patient in addition to tricuspid annuloplasty. Transmurality of the ablated atrial tissues was evaluated by electron microscopy. Heart function was assessed by transthoracic echocardiography. Cardiac rhythm was monitored by 24 h Holter at 3, 6, 12, 18, 24, and 48 months follow-up. 
All the AF was successfully eliminated in the ablation procedure without recurrence or other complications during hospitalization. The mean bypass and crossclamp times were similar in all the patients. The postoperative ventilator support time, the duration of stay in the ICU, and postoperative residence time were also not significantly different among the patients. Transmural atrial necrosis was detected in the ablated atrial tissues. Sinus rhythm maintenance was achieved at 3, 6, 12, 18, 24, and 48 months follow-up in all the patients. All valve protheses switched freely; no tricuspid regurgitation was observed. The results of the present study demonstrate that the CMP-IV is safe and effective in eliminating AF in dextrocardia patients concomitant with valvular surgery.

Surgical procedure and early postoperative period 
A mechanic MV replacement, tricuspid annuloplasty, and concomitant cryo-CMP-IV were performed simultaneously for the first patient. The second had undergone surgery of mechanic AV replacement, tricuspid annuloplasty, and concomitant cryo-CMP-IV. In the third patient, a biatrial CMP-IV procedure was performed simultaneously with mitral valvuloplasty and tricuspid annuloplasty. All operations were smooth, and each beating heart restored SR after resus...

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Cox-Maze IV Procedure Concomitant with Valvular Surgery In Situs Inversus Dextrocardia: A Single-Center Experience in China

We summarize the Cox-Maze IV procedure concomitant with valvular surgery performed in patients with situs inversus dextrocardia at this institution.

Atrial fibrillation (AF) is the most common cardiac arrhythmia. The use of ablation technologies made the Cox-Maze IV procedure (CMP-IV) technically ...

The efficacy and safety of the Cox-Maze IV procedure in dextrocardia are largely unknown. This protocol summarizes the procedure concomitantly with the vulvular surgery performed in three patients with dextrocardia. CMP4 is safe and effective in eliminating atrial fibrillation in dextrocardia.Moreover, the 3D-printed heart model helps preoperative simulating and planning for the surgical ablation of AF.Begin the surgical procedure for a median sternotomy by cannulating the superior and inferior vena cava to establish a mild hypothermic cardiopulmonary bypass, or CPB. After establishing the bypass, have an operator switch position from the right side to the left of the operating table. To achieve cardioplegic arrest, give an antegrade cold-blood cardioplegia intermittently from the aortic root.Then, perform the right atrial incision on the left side of the heart parallel to the atrioventricular sulcus. Ensure the left-sided left atriotomy is located parallelly under the interatrial groove. Place a retractor on the wall of the left atrium, or LA.For cryoablation, design a cryolesion set to replicate the mirror image of the CMP4 lesion set and maintain the duration of the LA cryoablation for each lesion at minus 60 degrees Celsius for two minutes.After confirming that the posterior LA box lesion is composed of an LA incision and cryolesion encircling the left and right pulmonary veins, apply a cryolesion line connecting the left superior pulmonary vein with the left atrial appendage, or LAA. Then, form an ice ball to mark the coronary sinus using cryoablation from the epicardium. When performing the mitral isthmus line, place the cryoprobe at the inferior aspect of the left atriotomy and direct the tube to the mitral annulus at the 8:00 position across the posterior LA and coronary sinus, as marked with the ice ball.Then, apply right-sided LAA amputation. After the LA cryoablation, sample a 4x8-millimeter tissue of the cryoablated LA for the electron microscopic examination and sample a large non-ablated tissue from the margin of the LA incision for the control test. Perform the prosthetic valve replacement surgery with a 27-millimeter mechanical mitral valve or a 23-millimeter mechanical atrium valve using a 2-0 polypropylene running suture.With the heart warm and beating, perform the right atrium cryoablation during the cardiopulmonary bypass for two minutes at minus 60 degrees Celsius for each ablation lesion. Create the linear cryoablation lines from the inferior aspect of the left-sided right atriotomy up onto the superior vena cava and down to the inferior vena cava. Make a tricuspid isthmus linear cryolesion from the mid-portion of the right atriotomy directed endocardially toward the tricuspid annulus at the 10:00 position, followed by a lateral cryolesion from the mid-portion of the right atriotomy up to the tip of the right atrial appendage, or RAA.Use the derived cardiac CT data to perform a 3D printing of the heart. During the operation, access the left atrium through the interatrial groove and extend the mitral isthmus lesion to the posterior mitral annulus. Then, ablate the coronary sinus in the endocardium and epicardium with a bipolar radiofrequency pen.Make the other lesions using bipolar radiofrequency clamps as described in the manuscript. When done, dissect the Marshall ligament and separate the left atrial appendage using an epicardial atrial clamp closure device. Then, use bipolar radiofrequency forceps to ablate the entire right atrial lesion sets.After resecting the A1-ruptured cordi, implant a single, flexible, artificial cord with 4-0 expanded polytetrafluoroethylene in situ and close the residual leak of the anterior commissure and the A2 leaflet cleft. To stabilize the annulus, implant a 32-millimeter rigid mitral ring, ensuring the coaptation height is nine millimeter. After deairing and closure of the interatrial sulcus incision, remove the aortic clamp and then make a longitudinal incision on the surface of the right atrium.Perform the tricuspid annuloplasty by implanting a 30-millimeter tricuspid ring in an upside-down and mirror image inversion manner using 2-0 polyester interrupted sutures. Remove the holder before fixation of the annuloplasty ring. Ensure that the patient's sinus rhythm is restored without any atrial ventricular block before cardiopulmonary bypass weaning.After the cardiac surgery, fix temporary epicardial pacing wires. In the postoperative course during the hospital stay, atrial fibrillation or no other complications were observed in any patient. After discharge, all the patients were followed up for one to three years, the 3, 6, 12, 18, 24 and 36 months follow-up in two patients, and at 3, 6, and 12 months follow-up in a patient was carried out.At three months after the surgery, all the heart functional capacities were improved to New York Heart Association Class I.The electron microscope of the samples collected from the cryoablated left atrium displayed tissue necrosis in the endocardium and muscularis. However, only edema and degeneration were observed in the epicardium and neighboring muscularis. The lesion set in dextrocardia should be designed to replicate the mirror image of the CMP4 lesion set.The tricuspid band must be implanted in an upside-down and mirror image inversion manner. Following this procedure, other methods, including chemical and physical ablation, can be performed using the biatrial lesion set of the CMP4 operation. However, cryoablation is the most recommended for valve surgery concomitant surgical ablation.The 3D-printed heart model displaying the spatial relationship between the specific ablation lines and the key anatomical references may help simulate and modify the CMP4 procedure, especially in patients with rare malformations.

cox-maze-iv-procedure-concomitant-with-valvular-surgery-situs

Research

JoVE Journal

Medicine

3.1K vues.  Central South University. L’efficacité et l’innocuité de la procédure Cox-Maze IV dans la dextrocardie sont largement inconnues. Ce protocole résume la procédure en concomitance avec la chirurgie vulvulaire réalisée chez trois patients atteints de dextrocardie. CMP4 est sûr et efficace pour éliminer la fibrillation auriculaire dans la dextrocardie.De plus, le modèle cardiaque imprimé en 3D aide à simuler et à planifier la simulation et la planification préopératoires de l’ablation chirurgic...

Vidéo: Cox-Maze IV Procedure Concomitant with Valvular Surgery In Situs Inversus Dextrocardia: A Single-Center Experience in China

We summarize the Cox-Maze IV procedure concomitant with valvular surgery performed in patients with situs inversus dextrocardia at this...

laparoscopic anterior right hepatectomy: a single-center experience

Laparoscopic Anterior Right Hepatectomy: A Single-Center Experience

Here we present a step-by-step protocol for conducting laparoscopic anterior right hepatectomy and compare its clinical effects and postoperative outcomes with those of conventional hepatectomies. Analysis of the data of 82 patients with hepatocellular carcinoma revealed that laparoscopic anterior right hepatectomy had better clinical outcomes and survival rates than the conventional...

visualizing the effects of a positive early experience, tactile stimulation, on dendritic morphology and synaptic connectivity with golgi-cox staining

Visualizing the Effects of a Positive Early Experience, Tactile Stimulation, on Dendritic Morphology and Synaptic Connectivity with Golgi-Cox Staining

This paper describes the procedures for tactile stimulation of rat pups and subsequent Golgi-Cox staining of neuronal morphology. Tactile stimulation is a positive experience that is administered in the perinatal period by stroking pups with a household duster. Golgi-cox staining is a reliable procedure permitting the visualization of entire...

procedure for electrode implantation surgery in mice

Procedure for Electrode Implantation Surgery in Mice

sudan iv staining: a procedure for staining lipid-laden atherosclerotic plaques in harvested mouse aortic arch

This video demonstrates a procedure using Sudan IV stain for staining the fatty deposits in the harvested mouse aortic arch to visualize the atherosclerotic plaques and determine the extent of...

Sudan IV Staining: A Procedure for Staining Lipid-Laden Atherosclerotic Plaques in Harvested Mouse Aortic Arch

a craniotomy surgery procedure for chronic brain imaging

A Craniotomy Surgery Procedure for Chronic Brain Imaging

This video and protocol demonstrate how to implant a glass-covered cranial window in rodents. These preparations can be used for chronic in vivo two-photon imaging of the neocortex over time scales of months. It may also be used for other types of imaging, including optical intrinsic signal...

improving iv insulin administration in a community hospital

Improving IV Insulin Administration in a Community Hospital

When compared to the previous paper protocol, implementation of a computerized glucose management system results in a substantial increase in blood glucose concentration measurements within the target range. Using a computerized glucose management system to monitor blood glucose levels, decreases in severe hypoglycemia (&lt;40 mg/dL), clinical hypoglycemia (&lt;70 mg/dL), and hyperglycemia (&gt;180 mg/dL) also can be...

Place a label on the back side of the pinning bed, and use a lead pencil to write down the mouse identification number. Transfer the aortic arch to the pinning bed, and add a drop of PBS to the tissue. Using a stereomicroscope, clean the aorta from the remaining periadventitial adipose tissue, and use Vannas scissors and Dumont forceps to gently peel away all of the surrounding adipose tissue without manipulating or damaging the...

laparoscopic common bile duct exploration in patients with a previous history of biliary tract surgery

Laparoscopic Common Bile Duct Exploration in Patients with a Previous History of Biliary Tract Surgery

Programmed surgery based on surgical methods and anatomical markers helps shorten the operation time, reduce complications, and improve the safety of the surgery. This study investigated and summarized the surgical methods and anatomic markers of laparoscopic common bile duct exploration in patients with a history of biliary tract...

testing drosophila olfaction with a y-maze assay

Testing Drosophila Olfaction with a Y-maze Assay

We present a choice test to reveal the influence of odorants on Drosophila behavior using a Y-maze...

Testing Drosophila Olfaction with a Y-maze Assay

learning modern laryngeal surgery in a dissection laboratory

Learning Modern Laryngeal Surgery in a Dissection Laboratory

The purpose of this paper is to illustrate how to organize a reproducible laboratory for laryngeal surgery on affordable and closely similar animal laryngeal models in order to improve anatomical and surgical knowledge and...

emergency undocking in robotic surgery: a simulation curriculum

Emergency Undocking in Robotic Surgery: A Simulation Curriculum

This training platform is designed to allow robotic surgeons to develop the skills necessary to lead an interprofessional team in emergency undocking of the robotic system. Training includes the utilization of the technology and equipment to perform an emergency undocking, as well as delineation of roles for such a...

sequencing of bacterial microflora in peripheral blood: our experience with hiv-infected patients

Sequencing of Bacterial Microflora in Peripheral Blood: our Experience with HIV-infected Patients

Our experiment will show how to perform a sequencing analysis of bacterial species translocating in peripheral blood of HIV positive patients.

op-ivm: combining in vitro maturation after oocyte retrieval with gynecological surgery

OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery

In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility...

technical considerations and approach to redo foregut surgery

Author Spotlight: Recent Advancements in Reoperative Foregut Surgery

Redo foregut surgery is associated with increased patient morbidity and presents a technical challenge for the surgeon. We describe our approach and considerations when performing a redo hiatal hernia repair to provide a guide for other surgeons and improve patient...

Reoperative Hiatal Hernia Repair

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isolation of valvular endothelial cells

Isolation of Valvular Endothelial Cells

We provide a method for isolating and culturing pure populations of heart valve endothelial cells (VEC).  VEC can be isolated from either side of the cusp or leaflet and immediately following, underlying interstitial cell (VIC) isolation is straightforward.

one-step metabolomics: carbohydrates, organic and amino acids quantified in a single procedure

One-step Metabolomics: Carbohydrates, Organic and Amino Acids Quantified in a Single Procedure

The urease method of sample preparation for GC/MS analysis of intermediary metabolites is presented by its inventor.  The method allows  one-step  follow-up of newborn screening for inborn errors by tandem mass spectrometry by quantifying carbohydrates, organic and amino acids all in a single...

chronic stress shifts effort-related choice behavior in a y-maze barrier task in mice

Chronic Stress Shifts Effort-Related Choice Behavior in a Y-Maze Barrier Task in Mice

The Y-maze barrier task is a behavior test that examines motivation to expend effort for reward. Here, we discuss testing multiple well-validated chronic stressors including chronic corticosterone and social defeat stress with this behavior, as well as the novel chronic non-discriminatory social defeat stress (CNSDS), which is effective in...

a personalized 3d-printed model for preoperative evaluation in thyroid surgery

A Personalized 3D-Printed Model for Preoperative Evaluation in Thyroid Surgery

Here, a new method of establishing a personalized 3D-printed model for preoperative evaluation of thyroid surgery is proposed. It is conducive to preoperative discussion, reducing the difficulty of thyroid...

a procedure for implanting organized arrays of microwires for single-unit recordings in awake, behaving animals

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

Implanting organized arrays of microwires for use in single-unit electrophysiological recordings presents a number of technical challenges.&#160;Methods for performing this technique and the equipment necessary are described.&#160;Also, the beneficial use of organized microwire arrays to record from distinct neural subregions with high spatial selectivity is...

The healthy gastrointestinal tract is physiologically colonized by a large variety of commensal microbes that influence the development 
	of the humoral and cellular mucosal immune system1,2. 
 Microbiota is shielded from the immune system via a strong mucosal barrier. Infections and antibiotics are known to alter both the normal 
	gastrointestinal tract barrier and the composition of resident bacteria, which may result in possible immune abnormalities3.
HIV causes a breach in the gastrointestinal barrier with progressive failure of mucosal immunity and leakage into the systemic circulation of bacterial bioproducts, such as lipopolysaccharide and 
	bacterial DNA fragments, which contribute to systemic immune activation4-7. Microbial translocation is implicated in HIV/AIDS immunopathogenesis and response to therapy 4,8.
We aimed to characterise the composition of bacteria translocating in peripheral blood of HIV-infected patients. To pursue our aim we set up a PCR reaction for the panbacteric 16S ribosomial gene 
	followed by a sequencing analysis.
Briefly, whole blood from both HIV-infected and healthy subjects is used. Given that healthy individuals present normal intestinal homeostasis no translocation of microflora is expected in 
	these patients. Following whole blood collection by venipuncture and plasma separation, DNA is extracted from plasma and used to perform a broad range PCR reaction for the panbacteric 
	16S ribosomial gene9. Following PCR product purification, cloning and sequencing analyses are performed.

Our experiment will show how to perform a sequencing analysis of bacterial species translocating in peripheral blood of HIV positive patients.

The healthy gastrointestinal tract is physiologically colonized by a large variety of commensal microbes that influence the development 
	of the humoral ...

Diabetes mellitus is a major independent risk factor for increased morbidity and mortality in the hospitalized patient, and elevated blood glucose concentrations, even in non-diabetic patients, predicts poor outcomes.1-4 The 2008 consensus statement by the American Association of Clinical Endocrinologists (AACE) and the American Diabetes Association (ADA) states that "hyperglycemia in hospitalized patients, irrespective of its cause, is unequivocally associated with adverse outcomes."5 It is important to recognize that hyperglycemia occurs in patients with known or undiagnosed diabetes as well as during acute illness in those with previously normal glucose tolerance.
The Normoglycemia in Intensive Care Evaluation-Survival Using Glucose Algorithm Regulation (NICE-SUGAR) study involved over six thousand adult intensive care unit (ICU) patients who were randomized to intensive glucose control or conventional glucose control.6 Surprisingly, this trial found that intensive glucose control increased the risk of mortality by 14% (odds ratio, 1.14; p=0.02). In addition, there was an increased prevalence of severe hypoglycemia in the intensive control group compared with the conventional control group (6.8% vs. 0.5%, respectively; p&lt;0.001). From this pivotal trial and two others,7,8 Wyoming Medical Center (WMC) realized the importance of controlling hyperglycemia in the hospitalized patient while avoiding the negative impact of resultant hypoglycemia. 
Despite multiple revisions of an IV insulin paper protocol, analysis of data from usage of the paper protocol at WMC shows that in terms of achieving normoglycemia while minimizing hypoglycemia, results were suboptimal. Therefore, through a systematical implementation plan, monitoring of patient blood glucose levels was switched from using a paper IV insulin protocol to a computerized glucose management system. By comparing blood glucose levels using the paper protocol to that of the computerized system, it was determined, that overall, the computerized glucose management system resulted in more rapid and tighter glucose control than the traditional paper protocol. Specifically, a substantial increase in the time spent within the target blood glucose concentration range, as well as a decrease in the prevalence of severe hypoglycemia (BG &lt; 40 mg/dL), clinical hypoglycemia (BG &lt; 70 mg/dL), and hyperglycemia (BG &gt; 180 mg/dL), was witnessed in the first five months after implementation of the computerized glucose management system. The computerized system achieved target concentrations in greater than 75% of all readings while minimizing the risk of hypoglycemia. The prevalence of hypoglycemia (BG &lt; 70 mg/dL) with the use of the computer glucose management system was well under 1%.

When compared to the previous paper protocol, implementation of a computerized glucose management system results in a substantial increase in blood glucose concentration measurements within the target range. Using a computerized glucose management system to monitor blood glucose levels, decreases in severe hypoglycemia (&lt;40 mg/dL), clinical hypoglycemia (&lt;70 mg/dL), and hyperglycemia (&gt;180 mg/dL) also can be observed.

Diabetes mellitus is a major independent risk factor for increased morbidity and mortality in the hospitalized patient, and elevated blood glucose ...

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in rodents, with detailed descriptions of the appropriate methods for anesthesia, the echocardiographic windows used, the imaging planes and probe orientations for image acquisition, the methods for data analysis, and the limitations of emerging technologies for the evaluation of cardiac and valvular function. Importantly, we also highlight several future areas of research in cardiac and heart valve imaging that may be leveraged to gain insights into the pathogenesis of valve disease in preclinical animal models. We propose that using a systematic approach to evaluating cardiac and heart valve function in mice can result in more robust and reproducible data, as well as facilitate the discovery of previously underappreciated phenotypes in genetically-altered and/or physiologically-stressed mice.

This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice.

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in ...

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model

Awake filling cystometry has been used for a long time to evaluate bladder function in freely moving mice, however, the specific methods used, vary among laboratories. The goal of this study was to describe the microsurgical procedure used to implant an intravesical tube and the experimental technique for recording urinary bladder pressure in an awake, freely moving mouse. In addition, experimental data is presented to show how surgery, as well as tubing type and size, affect lower urinary tract function and recording sensitivity. The effect of tube diameter on pressure recording was assessed in both polyethylene and polyurethane tubing with different internal diameters. Subsequently, the best performing tube from both materials was surgically implanted into the dome of the urinary bladder of male C57BL/6 mice. Twelve-hour, overnight micturition frequency was recorded in healthy, intact animals and animals 2, 3, 5, and 7 days post-surgery. At harvest, bladders were assessed for signs of swelling using gross observation and were subsequently processed for pathological analysis. The greatest extent of bladder swelling was observed on day 2 and 3, which correlated with behavioral voiding data showing significantly impaired bladder function. By day 5, bladder histology and voiding frequency had normalized. Based on the literature and evidence provided by our studies, we propose the following steps for in vivo recording of intravesical pressure and voided volume in an awake mouse: 1) Perform the surgery using an operating microscope and microsurgical tools, 2) Use polyethylene-10 tubing to minimize movement artifacts, and 3) Perform cystometry on post-operative day 5, when bladder swelling resolves.

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

Awake filling cystometry has been used for a long time to evaluate bladder function in freely moving mice, however, the specific methods used, vary among ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Establishing In Situ Closed Circuit Perfusion of Lower Abdominal Organs and Hind Limbs in Mice

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small size of mouse organs, ex vivo perfusion of bone, bladder, skin, prostate, and reproductive organs is challenging or not feasible. Here, we report for the first time an in situ lower body perfusion circuit in mice that includes the above tissues, but bypasses the main clearance organs (kidney, liver, and spleen). The circuit is established by cannulating the abdominal aorta and inferior vena cava above the iliac artery and vein and cauterizing peripheral blood vessels. Perfusion is performed via a peristaltic pump with perfusate flow maintained for up to 2 h. In situ staining with fluorescent lectin and Hoechst solution confirmed that the microvasculature was successfully perfused. This mouse model can be a very useful tool for studying pathological processes as well as mechanisms of drug delivery, migration/metastasis of circulating tumor cells into/from the tumor, and interactions of immune system with perfused organs and tissues.

A protocol is described for in situ perfusion of the mouse lower body, including the bladder, the prostate, sex organs, bone, muscle and foot skin.

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...

A Surgical Model of Heart Failure with Preserved Ejection Fraction in Tibetan Minipigs

More than half of heart failure (HF) cases are classified as heart failure with preserved ejection fraction (HFpEF) worldwide. Large animal models are limited for investigating the fundamental mechanisms of HFpEF and identifying potential therapeutic targets. This work provides a detailed description of the surgical procedure of descending aortic constriction (DAC) in Tibetan minipigs to establish a large animal model of HFpEF. This model used a precisely controlled constriction of the descending aorta to induce chronic pressure overload in the left ventricle. Echocardiography was used to evaluate the morphological and functional changes in the heart. After 12 weeks of DAC stress, the ventricular septum was hypertrophic, but the thickness of the posterior wall was significantly reduced, accompanied by dilation of the left ventricle. However, the LV ejection fraction of the model hearts was maintained at &gt;50% during the 12-week period. Furthermore, the DAC model displayed cardiac damage, including fibrosis, inflammation, and cardiomyocyte hypertrophy. Heart failure marker levels were significantly elevated in the DAC group. This DAC-induced HFpEF in minipigs is a powerful tool for investigating molecular mechanisms of this disease and for preclinical testing.

The present protocol describes a step-by-step procedure to establish a minipig model of heart failure with preserved ejection fraction using descending aortic constriction. The methods for evaluating cardiac morphology, histology, and function of this disease model are also presented.

More than half of heart failure (HF) cases are classified as heart failure with preserved ejection fraction (HFpEF) worldwide. Large animal models are ...

The anatomic structure of the surgical area of thyroid cancer is complex. It is very important to comprehensively and carefully evaluate the tumor location and its relation with the capsule, trachea, esophagus, nerves, and blood vessels before operation. This paper introduces an innovative 3D-printed model establishment method based on computerized tomography (CT) DICOM images. We established a personalized 3D-printed model of the cervical thyroid surgery field for each patient who needed thyroid surgery to help clinicians evaluate the key points and difficulties of the surgery and select the operation methods of key parts as a basis. The results showed that this model is conducive to preoperative discussion and the formulation of operation strategies. In particular, as a result of the clear display of the recurrent laryngeal nerve and parathyroid gland locations in the thyroid operation field, injury to them can be avoided during surgery, the difficulty of thyroid surgery reduced, and the incidence of postoperative hypoparathyroidism and complications related to recurrent laryngeal nerve injury reduced too. Moreover, this 3D-printed model is intuitive and aids communication for the signing of informed consent by patients before surgery.

Here, a new method of establishing a personalized 3D-printed model for preoperative evaluation of thyroid surgery is proposed. It is conducive to preoperative discussion, reducing the difficulty of thyroid surgery.

The anatomic structure of the surgical area of thyroid cancer is complex. It is very important to comprehensively and carefully evaluate the tumor ...

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs easily when performing another operation for laparoscopic common bile duct exploration (LCBDE), which was once considered a relative contraindication. In view of the limitations of the current surgical technique, this study summarized the surgical approaches and crucial anatomical landmarks for reoperation for LCBDE. Four general surgical approaches were proposed to expose the common bile duct, including the ligamentum teres hepatis approach, the anterior hepatic duodenal ligament approach, the right hepatic duodenal ligament approach, and the hybrid approach. Additionally, this study highlighted seven crucial anatomical landmarks: the parietal peritoneum, the gastrointestinal serosa, the ligamentum teres hepatis, the inferior margin of the liver, the gastric antrum, the duodenum, and the hepatic flexure of the colon, which were helpful to safely separate abdominal adhesions and expose the common bile duct. Moreover, to shorten the time of choledocholithotomy, a sequential method was innovatively applied for the removal of the stones in common bile duct. Mastering the above surgical approaches, including identifying crucial anatomical landmarks and adopting the sequential method will improve the safety of reoperation for LCBDE, shorten the operation time, promote the fast recovery of patients, reduce postoperative complications, and contribute to the popularization and application of this technique.

Programmed surgery based on surgical methods and anatomical markers helps shorten the operation time, reduce complications, and improve the safety of the surgery. This study investigated and summarized the surgical methods and anatomic markers of laparoscopic common bile duct exploration in patients with a history of biliary tract surgery.

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs ...