This study was supported by Clinical Research Training Program, the East Division of the First Affiliated Hospital of Sun Yat-Sen University (No.2019002, No.2019008), and the Foundation of National Health Commission of the People&#39;s Republic of China key laboratory of Male Reproduction and Genetics (No.KF202001).

The study was approved by the First Affiliated Hospital of Sun Yat-Sen University. Diagnostic criteria, surgical indications, and contraindications were in accordance with the Guidelines for Diagnosis and Treatment of Andrology and Expert Consensus of the Chinese Society of Andrology and the European Association of Urology Guidelines for Sexual and Reproductive Health. A patient would be excluded from this study if the female partner had medical conditions that affect fertility.
1. Instruments for operation
<ol>
	<li>Ensure the availability of sterilized instruments and other equipment mentioned in the Table of Materials.</li>
</ol>
2. Preparation for operation
<ol>
	<li>Prepare the bowel prior to surgery (by defecation).</li>
	<li>Prepare the perineal skin before surgery (by shaving off the hair).</li>
	<li>Administer intramuscular scopolamine (20 mg) or atropine (0.5 mg) and an intravenous infusion of cefazolin sodium (1 g) with 100 mL of 0.9% sodium chloride solution 30 min before surgery.</li>
	<li>Place the patient in the supine position on the operating table after combined spinal-epidural anesthesia.</li>
	<li>Disinfect the surgical area with 1% iodophor and cover it with surgical towels.</li>
</ol>
3. Vessel-sparing modified single-armed LIVE
<ol>
	<li>Insert a 16 French or 16Fr Foley catheter, and mark the incision sites with a skin marker.</li>
	<li>Deliver the testis through a 3-4 cm vertical scrotal incision.</li>
	<li>Expose the vas deferens with a vas-fixation clamp next to the spermatic cord near the testis. Pass a vascular sling through the space between the vas deferens and the vas deferens vasculature. Apply traction to the vascular sling, separate the connective tissue with microhemostatic forceps under the operating microscope, and cut with an electric knife. Carefully dissociate deferential vessels with the microhemostatic forceps by 1 cm from the vas deferens under the operating microscope (Figure 1).</li>
	<li>After hemisecting the vas deferens with a knife (Figure 2A), confirm the patency of the vas deferens by injection of diluted methylene blue with a 24 G irrigating needle connected to a 1 mL syringe and observation of dye in the urine, or by injection of 0.9% sodium chloride solution with no resistance or reflux (Figure 2B).</li>
	<li>Separate the adhesion between the tunica vaginalis and the testis with microhemostatic forceps, and cut with an electric knife after opening the tunica vaginalis. Examine the epididymis under the operating microscope, and select the site of dilated epididymal tubule for anastomosis. Cut a 5 mm diameter piece of the epididymal tunic using ophthalmic scissors at this position.</li>
	<li>Completely transect the vas deferens with a knife, and ligate the broken end of the vas deferens near the epididymis with silk, braided, nonabsorbable suture. Perforate the tunica vaginalis using hemostatic forceps and pass the isolated part of the vas deferens through the tunnel to reach the anastomosis site (Figure 3A). Fix the vas deferens and epididymis tunic using two interrupted 8-0 polypropylene sutures, and ensure the vas deferens is not twisted at the same time. Use microscopic bipolar coagulation to stop the vas deferens from bleeding to keep the operative field clear (Figure 3B).</li>
	<li>After marking four suture sites on the vas deferens (equidistant distribution on the section of the vas deferens, Figure 4A), perform this modified single-armed suture technique for LIVE using two single-armed 10-0 polypropylene sutures7. Pass two needles through the inferior points of the vasal mucosal layer separately in an outside-in fashion (Figure 4A: a1, b1), using a microneedle holder to slightly dilate the vasal lumen and accurately control the needle under the microscope. Avoid hooking the needle into the back wall of the lumen (Figure 4B). Move the two needles in parallel and longitudinally through the same epididymal tubule.</li>
	<li>Place two needles in the epididymal tubule, opened longitudinally between the two needles using a 15&#176; ophthalmic knife (Figure 5A). Aspirate the epididymal fluid flowing from the incision in the tubule with a 24 G irrigating needle connected to a 1 mL syringe, and hand it to an examiner to check for sperm (Figure 5B).</li>
	<li>Gently pull out two needles in the epididymal tubule separately and pass them through the superior points (Figure 6A: a2, b2) of the vasal mucosal layer in an inside-out fashion. Suture the adventitia of the vas deferens and the epididymal tunic with an 8-0 polypropylene suture to reduce tension before intussusception of the epididymal tubule into the vas deferens can be performed (Figure 6B).</li>
	<li>Suture the muscularis edge of the vas deferens and the epididymal tunic using 10-12 interrupted sutures of 9-0 polypropylene (Figure 7).</li>
</ol>
4. Postoperative care
<ol>
	<li>Ensure that the patient waits for 6 h after the operation before eating, takes bed rest for 3 days, and avoids standing or walking.</li>
	<li>Continue intravenous antibiotics (cefazolin sodium, 1 g with 100 mL of 0.9% sodium chloride solution) after the surgery to avoid genitourinary tract infection or epididymitis.</li>
</ol>

<ol>
	<li>Salonia, , 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=European+Association+of+urology+guidelines+on+sexual+and+reproductive+health-2021+update:+male+sexual+dysfunction.">European Association of urology guidelines on sexual and reproductive health-2021 update: male sexual dysfunction.</a> European Urology. 80 (3), 333-357 (2021).</li><li>Chen, X. 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=Microsurgical+vasoepididymostomy+for+patients+with+infectious+obstructive+azoospermia:+cause,+outcome,+and+associated+factors.">Microsurgical vasoepididymostomy for patients with infectious obstructive azoospermia: cause, outcome, and associated factors.</a> Asian Jorunal of Andrology. 18 (5), 759-762 (2016).</li><li>Han, H., Liu, S., Zhou, X. G., Tian, L., Zhang, X. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aetiology+of+obstructive+azoospermia+in+Chinese+infertility+patients.">Aetiology of obstructive azoospermia in Chinese infertility patients.</a> Andrologia. 48 (7), 761-764 (2016).</li><li>Chan, P. T. K., Goldstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+reconstruction+of+pre-epididymal+obstructive+azoospermia.">Microsurgical reconstruction of pre-epididymal obstructive azoospermia.</a> Journal of Urology. 163, 258 (2000).</li><li>Practice Committee of the American Society for Reproductive Medicine in collaboration with the Society for Male Reproduction and Urology. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+management+of+obstructive+azoospermia:+a+committee+opinion.">The management of obstructive azoospermia: a committee opinion.</a> Fertility and Sterility. 111 (5), 873-880 (2019).</li><li>Chan, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+evolution+and+refinement+of+vasoepididymostomy+techniques.">The evolution and refinement of vasoepididymostomy techniques.</a> Asian Jorunal of Andrology. 15 (1), 49-55 (2013).</li><li>Lyu, K. L., 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+novel+experience+of+deferential+vessel-sparing+microsurgical+vasoepididymostomy.">A novel experience of deferential vessel-sparing microsurgical vasoepididymostomy.</a> Asian Jorunal of Andrology. 20 (6), 576-580 (2018).</li><li>Fantus, R. J., Halpern, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vasovasostomy+and+vasoepididymostomy:+indications,+operative+technique,+and+outcomes.">Vasovasostomy and vasoepididymostomy: indications, operative technique, and outcomes.</a> Fertility and Sterility. 115 (6), 1384-1392 (2021).</li><li>Baker, K., Jr, S. a. b. a. n. e. g. h. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Obstructive+azoospermia:+reconstructive+techniques+and+results.">Obstructive azoospermia: reconstructive techniques and results.</a> Clinics. 68, 61-73 (2013).</li><li>Monoski, M. A., Schiff, J., Li, P. S., Chan, P. T., Goldstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innovative+single-armed+suture+technique+for+microsurgical+vasoepididymostomy.">Innovative single-armed suture technique for microsurgical vasoepididymostomy.</a> Urology. 69 (4), 800-804 (2007).</li><li>Zhao, L., 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+modified+single-armed+technique+for+microsurgical+vasoepididymostomy.">A modified single-armed technique for microsurgical vasoepididymostomy.</a> Asian Journal of Andrology. 15 (1), 79-82 (2013).</li><li>Zhao, L., 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=Retrospective+analysis+of+early+outcomes+after+a+single-armed+suture+technique+for+microsurgical+intussusception+vasoepididymostomy.">Retrospective analysis of early outcomes after a single-armed suture technique for microsurgical intussusception vasoepididymostomy.</a> Andrology. 3 (6), 1150-1153 (2015).</li><li>Harrison, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+distribution+of+the+vasal+and+cremasteric+arteries+to+the+testis+and+their+functional+importance.">The distribution of the vasal and cremasteric arteries to the testis and their functional importance.</a> Journal of Anatomy. 83, 267-282 (1949).</li><li>Raman, J. D., Goldstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+characterization+of+arterial+vasculature+in+spermatic+cord.">Intraoperative characterization of arterial vasculature in spermatic cord.</a> Urology. 64 (3), 561-564 (2004).</li><li>Mostafa, T., Labib, I., El-Khayat, Y., El-Rahman El-Shahat , A., Gadallah, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+testicular+arterial+supply:+gross+anatomy,+corrosion+cast,+and+radiologic+study.">Human testicular arterial supply: gross anatomy, corrosion cast, and radiologic study.</a> Fertility and Sterility. 90 (6), 2226-2230 (2008).</li><li>Student, V., Z&#225;tura, F., Scheinar, J., Vrtal, R., Vr&#225;na, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testicle+hemodynamics+in+patients+after+laparoscopic+varicocelectomy+evaluated+using+color+Doppler+sonography.">Testicle hemodynamics in patients after laparoscopic varicocelectomy evaluated using color Doppler sonography.</a> European Urology. 33 (1), 91-93 (1998).</li><li>Matsuda, T., Horii, Y., Yoshida, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Should+the+testicular+artery+be+preserved+at+varicocelectomy.">Should the testicular artery be preserved at varicocelectomy.</a> The Journal of Urology. 149 (5), 1357-1360 (1993).</li><li>Li, P., 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=Vasal+vessel-sparing+microsurgical+single-armed+vasoepididymostomy+to+epididymal+obstructive+azoospermia:+A+retrospective+control+study.">Vasal vessel-sparing microsurgical single-armed vasoepididymostomy to epididymal obstructive azoospermia: A retrospective control study.</a> Andrologia. 53 (8), 14133 (2021).</li></ol>

The authors have no conflicts of interest to disclose.

Genitourinary tract infections and epididymitis are common causes of epididymal OA. VE has become an important method to treat epididymal OA and has been applied in clinics since 20004. Anastomosis of the vas deferens and epididymis is performed with double-needle sutures without preserving vessels of the vas deferens in the classic MVE6,8,9. Because double-needle sutures are expensive and not readily ava...

The number of infertile couples has been increasing annually; OA occurs in 20%-40% of azoospermia cases in men of reproductive age1. Epididymal obstruction accounts for approximately 30% of OA cases and is one of the most common obstruction sites. However, this proportion may be higher in China2,3. The treatment for OA varies depending on the site of the obstruction. The common causes of OA include vasectomy, genitourinary tract infection, genitourinary tuberculosis, iatrogenic injury, and idiopathic obstruction. The etiology of OA in China is mostly epididymal obstruction caused by genitourinary tract infection or epididymitis, while vasectomy is the most common etiology in Western countries2,3. The two types of obstructions require slightly different surgical approaches.
Microsurgical vasoepididymostomy (MVE) has become an important method for treating epididymal OA since 20004. MVE is the most challenging operation in male microsurgery, including microsurgical end-to-end single-tubule anastomosis, end-to-side anastomosis, triangulation, tubular invagination, and tubular intussusception techniques5. Longitudinal intussusception vasoepididymostomy (LIVE) is more advantageous because of the wider opening of the epididymal tubule6,7,8. Based on the characteristics of this case (presented here) in China, an improved, vessel-sparing, modified, single-armed suture LIVE technique was proposed based on a modified single-armed suture MVE technique. This technique not only enables vasoepididymostomy (VE) to be performed in areas where double-needle sutures are not readily available, but also preserves the vasculature of the vas deferens and maintains the normal physiological structure.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% sodium chloride solution</td>
			<td>Guangdong Otsuka Pharmaceutical Co. LTD</td>
			<td>21M1204</td>
			<td>Dilute antibiotics, irrigate.</td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>Kindly Medical, Shanghai</td>
			<td>K20210826</td>
			<td>inject diluted methylene blue or 0.9% sodium chloride solution</td>
		</tr>
		<tr>
			<td>1% iodophor</td>
			<td>Guangzhou Qingfeng Disinfection Products Co., LTD</td>
			<td>Q/QFXD2</td>
			<td>Disinfect the surgical area.</td>
		</tr>
		<tr>
			<td>10-0 polypropylene sutures</td>
			<td>Ethicon, LLC</td>
			<td>REBBES</td>
			<td>Used when anastomosing.</td>
		</tr>
		<tr>
			<td>3-0 polyglactin 910 sutures</td>
			<td>Ethicon, LLC</td>
			<td>RGMCLH</td>
			<td>Suture skin incisions at the end of surgery.</td>
		</tr>
		<tr>
			<td>5-0 polyglactin 910 sutures</td>
			<td>Ethicon, LLC</td>
			<td>RBMMPQ</td>
			<td>Suture skin incisions at the end of surgery.</td>
		</tr>
		<tr>
			<td>8-0 polypropylene sutures</td>
			<td>Ethicon, LLC</td>
			<td>RDBBLS</td>
			<td>Used when anastomosing.</td>
		</tr>
		<tr>
			<td>9-0 polypropylene sutures</td>
			<td>Ethicon, LLC</td>
			<td>RABDTE</td>
			<td>Used when anastomosing.</td>
		</tr>
		<tr>
			<td>F16 urinary catheter</td>
			<td>Well Lead Medical, Guangzhou</td>
			<td>20190612</td>
			<td>Drainage of urine due to long operation time.</td>
		</tr>
		<tr>
			<td>micro haemostatic forceps</td>
			<td>Shanghai Surgical Instrument Factory</td>
			<td>W40350</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>micro scissors</td>
			<td>Cheng-He,NingBo</td>
			<td>HC-A008</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>micro tweezers</td>
			<td>Cheng-He,NingBo</td>
			<td>HC-A002</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>microneedle holder</td>
			<td>Cheng-He,NingBo</td>
			<td>HC-GN006</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>ophthalmic scissors</td>
			<td>Shanghai Surgical Instrument Factory</td>
			<td>Y00040</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>polyglactin 910 sutures</td>
			<td>Ethicon, LLC</td>
			<td>RBMMPQ</td>
			<td>Suture skin incisions at the end of surgery.</td>
		</tr>
		<tr>
			<td>silk braided non-absorbable suture</td>
			<td>Ethicon, LLC</td>
			<td>SB84G</td>
			<td>ligate the broken end of the vas deferens</td>
		</tr>
		<tr>
			<td>skin marker</td>
			<td>Medplus Inc.</td>
			<td>21120206</td>
			<td>Mark surgical incisions and suture sites.</td>
		</tr>
		<tr>
			<td>surgical microscope Carl Zeiss S88</td>
			<td>Carl Zeiss</td>
			<td>Carl Zeiss S88</td>
			<td>Enlarge your field of vision during surgery.</td>
		</tr>
		<tr>
			<td>vas-fixation clamp</td>
			<td>Shanghai Surgical Instrument Factory</td>
			<td>JCZ220</td>
			<td>Used in surgical procedures</td>
		</tr>
	</tbody>
</table>

vessel-sparing microsurgical longitudinal intussusception vasoepididymostomy to treat epididymal obstructive azoospermia

The epididymis is a common site of obstruction in obstructive azoospermia (OA). Vasoepididymostomy has become an important method for the treatment of epididymal OA since 2000. There are two challenges in classic microscopic vasoepididymostomy. First, anastomosis of the vas deferens and epididymis is performed with double-needle sutures. However, there is a lack of good-quality and cost-effective double-needle sutures in China, which leads to increased difficulty and poor success rates of anastomosis. Second, the separation of the vas deferens does not retain vasculature, although the vas deferens vasculature plays an important role in the blood supply to the vas deferens, epididymis, and testis. This affects the blood supply to the anastomotic area and epididymis.
Therefore, this team has made innovative improvements to address these problems. Good-quality, cost-effective, single-needle sutures, which are easy to purchase in China and other countries, were used in microsurgical longitudinal intussusception vasoepididymostomy. This can optimize the operation procedure and shorten the operation time while ensuring the success rate of the anastomosis. The surgical method of preserving the vas deferens vessels was innovatively proposed because the etiology of epididymal OA is mostly inflammatory in China. The protection of the blood supply to the vas deferens and epididymis is maximized using microsurgical forceps to separate and protect the vasculature. Patency reached 81.7% in the postoperative follow-up, indicating a better surgical treatment effect.

A study included 92 men who were diagnosed with azoospermia secondary to epididymal obstruction in the First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China, and who underwent operation between January 2017 and December 2018. The average age of the 92 males was 30.77 &#177; 5.38 years (range: 20-47 years) (Table 1). All men underwent the bilateral vessel-sparing, modified, single-armed technique for LIVE, and the mean operation time was 223.59 &#177; 31.73 min. No postoperative complicati...

Watch this Scientific Journal Video about Vessel-Sparing Microsurgical Longitudinal Intussusception Vasoepididymostomy to Treat Epididymal Obstructive Azoospermia at JoVE.com

Vessel-Sparing Microsurgical Longitudinal Intussusception Vasoepididymostomy to Treat Epididymal Obstructive Azoospermia

The present protocol describes a vessel-sparing, longitudinal intussusception vasoepididymostomy using readily available single-needle sutures in China, a safe and effective procedure, which may improve patients' patency and natural pregnancy rates.

The epididymis is a common site of obstruction in obstructive azoospermia (OA). Vasoepididymostomy has become an important method for the treatment of ...

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3.6K Views.  The East Division of the First Affiliated Hospital of Sun Yat-Sen University. The present protocol describes a vessel-sparing, longitudinal intussusception vasoepididymostomy using readily available single-needle sutures in China, a safe and effective procedure, which may improve patients' patency and natural pregnancy rates.

Video: Vessel-Sparing Microsurgical Longitudinal Intussusception Vasoepididymostomy to Treat Epididymal Obstructive Azoospermia

Dual-phase Cone-beam Computed Tomography to See, Reach, and Treat Hepatocellular Carcinoma during Drug-eluting Beads Transarterial Chemo-embolization

The advent of cone-beam computed tomography (CBCT) in the angiography suite has been revolutionary in interventional radiology. CBCT offers 3 dimensional&#160;(3D) diagnostic imaging in the interventional suite and can enhance minimally-invasive therapy beyond the limitations of 2D angiography alone. The role of CBCT has been recognized in transarterial chemo-embolization (TACE) treatment of hepatocellular carcinoma (HCC). The recent introduction of a CBCT technique: dual-phase CBCT (DP-CBCT) improves intra-arterial HCC treatment with drug-eluting beads (DEB-TACE). DP-CBCT can be used to localize liver tumors with the diagnostic accuracy of multi-phasic multidetector computed tomography (M-MDCT) and contrast enhanced magnetic resonance imaging (CE-MRI) (See the tumor), to guide intra-arterially guidewire and microcatheter to the desired location for selective therapy (Reach the tumor), and to evaluate treatment success during the procedure (Treat the tumor). The purpose of this manuscript is to illustrate how DP-CBCT is used in DEB-TACE to see, reach, and treat HCC.

Dual-phase cone-beam computed tomography (DP-CBCT) is a useful intraprocedural imaging technique for transarterial chemo-embolization treatment with drug-eluting beads of hepatocellular carcinoma. DP-CBCT has been used to perform three major steps in oncologic interventional radiology: tumor localization (see), navigation and intraprocedural catheter guidance (reach), and intraprocedural evaluation of treatment success (treat).

The advent of cone-beam computed tomography (CBCT) in the angiography suite has been revolutionary in interventional radiology. CBCT offers 3 ...

The Helsinki Rat Microsurgical Sidewall Aneurysm Model

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced into clinical practice. Furthermore, experimental models are needed to elucidate the complex aneurysm biology leading to rupture of saccular aneurysms.
Several different kinds of experimental models for saccular aneurysms have been established in different species. Many of them, however, require special skills, expensive equipment, or special environments, which limits their widespread use. A simple, robust, and inexpensive experimental model is needed as a standardized tool that can be used in a standardized manner in various institutions.
The microsurgical rat abdominal aortic sidewall aneurysm model combines the possibility to study both novel endovascular treatment strategies and the molecular basis of aneurysm biology in a standardized and inexpensive manner. Standardized grafts by means of shape, size, and geometry are harvested from a donor rat&#39;s descending thoracic aorta and then transplanted to a syngenic recipient rat. The aneurysms are sutured end-to-side with continuous or interrupted 9-0 nylon sutures to the infrarenal abdominal aorta.
We present step-by-step procedural instructions, information on necessary equipment, and discuss important anatomical and surgical details for successful microsurgical creation of an abdominal aortic sidewall aneurysm in the rat.

Microsurgical sidewall aneurysms in rats are created by end-to-side anastomosis of an aortic graft to the abdominal aorta. We present step-by-step instructions and discuss anatomical and surgical details for successful experimental saccular aneurysm creation.

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced ...

Step By Step: Microsurgical training method combining two nonliving animal models

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly laboratory rats. This method although extremely valuable can be economically demanding both for the surgeon and the sponsoring institution; it also requires special training facilities that may not always be available or accessible. Furthermore ethical concerns can limit the use of living animals for training purposes. Alternative training methods, such as inert tubes and gloves have not gained popularity among surgeons since they do not offer an experience similar to that of a clinical situation. Non-living animal models include the use of chicken thighs and wings; they offer a practice experience that resembles a clinical situation to a considerable extent. This type of training is relatively cheap and easily available. The microscope and instruments required can be acquired over the internet, and the chicken pieces can be bought at the local supermarket.
This approach allows a motivated trainee to rehearse different types of surgical techniques several times at a reasonable expense, helping to develop or maintain his surgical expertise if more complex facilities are not available. On the current manuscript we describe how to setup a small practice station, how to dissect the specimens, and how to practice both with the chicken thighs and with the chicken wings in a progressive fashion. This approach takes advantage on the versatility of the chicken thigh model and the small size of the chicken wing Brachial artery.

Here we describe how to set up a small microsurgical practice station and a simple and inexpensive method for the training of microsurgery with non-living animal models.

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular plasticity or damage occurring overtime and in relation to neuronal activity or blood flow. In vivo two-photon microscopy allows the study of the structural and functional plasticity of large cellular units in the living brain. In particular, the thinned-skull window preparation allows the visualization of cortical regions of interest (ROI) without inducing significant brain inflammation. Repetitive imaging sessions of cortical ROI are feasible, providing the characterization of disease hallmarks over time during the progression of numerous CNS diseases. This technique accessing the pial structures within 250 &#956;m of the brain relies on the detection of fluorescent probes encoded by genetic cellular markers and/or vital dyes. The latter (e.g., fluorescent dextrans) are used to map the luminal compartment of cerebrovascular structures. Germane to the protocol described herein is the use of an in vivo marker of amyloid deposits, Methoxy-O4, to assess Alzheimer&#39;s disease (AD) progression. We also describe the post-acquisition image processing used to track vascular changes and amyloid depositions. While focusing presently on a model of AD, the described protocol is relevant to other CNS disorders where pathological cerebrovascular changes occur.

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

This manuscript describes a procedure to track the remodeling of the cerebrovasculature during amyloid plaque accumulation in vivo using longitudinal two-photon microscopy. A thinned-skull preparation enables the visualization of fluorescent dyes to assess the progression of cerebrovascular damage in a mouse model of Alzheimer&#39;s disease.

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

Development of Recombinant Proteins to Treat Chronic Pain

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates for treating debilitating pain conditions due to their capacity to regulate aberrant neuro-immune interactions. However, physiologically they work in a network of various cytokines, and therefore their therapeutic effect may not be optimal when used as stand-alone drugs. To overcome this limitation, we developed a fusion protein of the anti-inflammatory cytokines IL4 and IL10. Here, we describe the methods for production and quality control of IL4-10 recombinant fusion protein and we test the effectiveness of the IL4-10 fusion protein to resolve pain in a mouse model of persistent inflammatory pain.

In this paper we provide details for the methods of production and quality control of an IL4-10 recombinant fusion protein. We also show how to test the effectiveness of this protein to resolve pain in a mouse model of inflammatory pain.

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates ...

Vessel-sparing Excision and Primary Anastomosis

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short bulbar or posterior urethral strictures, urethroplasty by excision and primary anastomosis (EPA) is recommended. As EPA only requires the excision of the narrowed segment and the surrounding spongiofibrosis, a full-thickness transection of the corpus spongiosum, as performed in the traditional transecting EPA (tEPA), is usually unnecessary. Jordan et al. introduced the idea of a vessel-sparing approach in 2007, aiming to reduce surgical trauma, especially to the dual arterial blood supply of the urethra, and, thus, potentially reducing the risk of postoperative erectile dysfunction or glans ischemia. This approach could also be beneficial for subsequent urethral interventions such as redo urethroplasty using a free graft, in which a well-vascularized graft bed is imperative. Nevertheless, these potential benefits are only assumptions as prospective studies comparing the functional outcome of both techniques with validated questionnaires are currently lacking. Moreover, vessel-sparing EPA (vsEPA) should at least be able to provide similar surgical outcomes as tEPA. The aim of this paper is to give an elaborate, step-by-step overview of how to manage patients with isolated short bulbar or posterior urethral strictures with vsEPA. The main objective of this manuscript is to outline the surgical technique and to report the representative surgical outcome. A total of 117 patients were managed according to the described protocol. The analysis was performed on the entire patient cohort and on the bulbar (n = 91) and posterior (n = 26) vsEPA group separately. Success rates were 93.4% and 88.5% for the bulbar and posterior vsEPA, respectively. To conclude, vsEPA, as outlined in the protocol, provides excellent success rates with low complication rates for isolated short bulbar and posterior urethral strictures.

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

A Minimally Invasive Method for Intratracheal Instillation of Drugs in Neonatal Rodents to Treat Lung Disease

Treatment of neonatal rodent with drugs instilled directly into the trachea could serve as a valuable tool to study the impact of a locally administered drug. This has direct translational impact because surfactant and drugs are administered locally into the lungs. Though the literature has many publications describing minimally invasive transoral intubation of adult mice and rats in therapeutic experiments, this approach in neonatal rat pups is lacking. The small size of orotracheal region/pharynx in the pups makes visualization of laryngeal lumen (vocal cords) difficult, contributing to the variable success rate of intratracheal drug delivery. We hereby demonstrate effective oral intubation of neonatal rat pup - a technique that is non-traumatic and minimally-invasive, so that it can be used for serial administration of drugs. We used an operating otoscope with an illumination system and a magnifying lens to visualize the tracheal opening of the rat neonates. The drug is then instilled using a 1 mL syringe connected to a pipette tip. The accuracy of the delivery method was demonstrated using Evans blue dye administration. This method is easy to get trained in and could serve as an effective way to instill drugs into trachea. This method could also be used for administration of inoculum or agents to simulate disease conditions in animals and, also, for cell-based treatment strategies for various lung diseases.

This technique of instilling drugs directly into the trachea of neonatal rodents is important in studying the impact of locally administered drugs or biologicals on neonatal lung diseases. Additionally, this method can also be used for inducing lung injury in animal models.

Treatment of neonatal rodent with drugs instilled directly into the trachea could serve as a valuable tool to study the impact of a locally administered ...

Fat-Covered Islet Transplantation Using Epididymal White Adipose Tissue

Islet transplantation is a cellular replacement therapy for severe diabetes mellitus. The intraperitoneal cavity is typically the transplant site for this procedure. However, intraperitoneal islet transplantation has some limitations, including poor transplant efficacy, difficult graft detection ability, and a lack of graftectomy capability for post-transplant analysis. In this paper, "fat-covered islet transplantation", an intraperitoneal islet transplantation method that utilizes epididymal white adipose tissue, is used to assess the therapeutic effects of bioengineered islets. The simplicity of the method lies in the seeding of islets onto epididymal white adipose tissue and using the tissue to cover the islets. While this method can be categorized as an intraperitoneal islet transplantation technique, it shares characteristics with intra-adipose tissue islet transplantation. The fat-covered islet transplantation method demonstrates more robust therapeutic effects than intra-adipose tissue islet transplantation, however, including the improvement of blood glucose and plasma insulin levels and the potential for graft removal. We recommend the adoption of this method for assessing the mechanisms of islet engraftment into white adipose tissue and the therapeutic effects of bioengineered islets.

This fat-covered islet transplantation method is suitable for the detection of engrafted islets in the intraperitoneal cavity. Notably, it does not require the use of biobinding agents or suturing.

Islet transplantation is a cellular replacement therapy for severe diabetes mellitus. The intraperitoneal cavity is typically the transplant site for this ...

A Modified Vessel-Sparing Microsurgical Vasoepididymostomy

Microsurgical vasoepididymostomy (MVE) is the main surgical treatment for epididymal obstruction. The vasal vessels are ligated during MVE. However, preserving the vasal vessels during MVE might better simulate the normal physiological structure and be meaningful for patients who have undergone varicocelectomy. Nevertheless, preserving the vasal vessels might elevate the risk of increasing the tension of anastomosis, affecting the patency rate and leading to delayed postoperative bleeding. Therefore, we developed a novel vessel-sparing MVE to make it safer. Here is a summary of the improvements to the procedure. 1) The retrograde dissociation of the vasal vessels on the proximal testicular side was adopted as the main method, and the anterograde dissociation of the vasal vessels on the distal testicular side was adopted as a supplement to dissociate the vasal vessels to be preserved. This improvement ensures the blood supply to the vas deferens that will be used for anastomosis and also provides longer vasal vessels, which reduces the tension of anastomosis. 2) By fixing the vas deferens to be anastomosed and the broken end of the vas deferens, the free vasal vessels get fixed, which resolves the problem of transmission of vas tension to the vasal vessels and reduces the risk of vasal vessel hemorrhage. 3) Dissociation of the vas deferens after opening the tunica vaginalis increases the mobilization of the vas deferens, which also makes the new procedure easier to complete. The evaluation of the outcomes of this new procedure showed that no significant postoperative complications occurred in the patients, and the patency rate was no different from that of the conventional procedure. Therefore, this new, improved procedure can be considered safe, with satisfactory postoperative results.

Here, we present a protocol to preserve the vasal vessels in microsurgical vasoepididymostomy. The surgical security is enhanced by preserving the vasal vessels using a retrograde-anterograde dissociation and fixing the vasal vessels.

Microsurgical vasoepididymostomy (MVE) is the main surgical treatment for epididymal obstruction. The vasal vessels are ligated during MVE. However, ...