Name: a modified technique for arteriovenous fistula construction in rabbits
Uploaded: 2023-02-10T13:33:00
Description: Watch this Scientific Journal Video about A Modified Technique for Arteriovenous Fistula Construction in Rabbits at JoVE.com

This study was supported by grants from the Suzhou Science and Technology Plan Project (SYS2020077), Suzhou High-tech Zone Medical and Health Science and Technology Plan Project (2020z001), Suzhou Science and Technology Development Plan Project-Medical and Health Science and Technology Innovation (SYK2021030), Nanjing Medical University Science and Technology Development Fund-General Project (NMUB20210253), Suzhou Science and Technology Bureau of the application of the basic research project (No.SYSD2019205, No.SYS2020119), Jiangsu Province Traditional Chinese Medicine Science and Technology Development Plan Project (No.MS2021098), the Ministry of Education Industry-University Cooperation Collaborative Education Project (No. 202102242003),&#160;the Sixth &#34;333 High-level Talent Cultivation&#34; Project in Jiangsu Province, Suzhou Science and Technology City Hospital 2022 Hospital-level Pre-research Fund Project (SZKJCYY2022014), and Suzhou &#34;KeJiaoXingWei&#34; Youth Science and Technology Project (KJXW2022086).

The experimental procedures using laboratory animals were approved by the Experimental Animal Welfare Ethics Committee of Nanjing Medical University. New Zealand rabbits aged 10 months (of both sexes; body weight, 3.18 &#177; 0.24 kg) were used for this study. The animals were obtained from a commercial source (see Table of Materials).
1. Animal preparation
<ol>
	<li>Anesthetize the rabbits using a mixed intravenous injection of tiletamine hydrochloride and ...

<ol>
	<li>Lok, C. E., 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=KDOQI+Clinical+Practice+Guideline+for+Vascular+Access:+2019+update.">KDOQI Clinical Practice Guideline for Vascular Access: 2019 update.</a> American Journal of Kidney Diseases. 75, 1 (2020).</li><li>Schmidli, J., 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=Editor's+choice+-+Vascular+access:+2018+Clinical+Practice+Guidelines+of+the+European+Society+for+Vascular+Surgery+(ESVS).">Editor's choice - Vascular access: 2018 Clinical Practice Guidelines of the European Society for Vascular Surgery (ESVS).</a> European Journal of Vascular and Endovascular Surgery. 55 (6), 757-818 (2018).</li><li>Grogan, J., 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=Frequency+of+critical+stenosis+in+primary+arteriovenous+fistulae+before+hemodialysis+access:+Should+duplex+ultrasound+surveillance+be+the+standard+of+care.">Frequency of critical stenosis in primary arteriovenous fistulae before hemodialysis access: Should duplex ultrasound surveillance be the standard of care.</a> Journal of Vascular Surgery. 41 (6), 1000-1006 (2005).</li><li>Swinnen, J., Lean, T. K., Allen, R., Burgess, D., Mohan, I. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Juxta-anastomotic+stenting+with+aggressive+angioplasty+will+salvage+the+native+radiocephalic+fistula+for+dialysis.">Juxta-anastomotic stenting with aggressive angioplasty will salvage the native radiocephalic fistula for dialysis.</a> Journal of Vascular Surgery. 61 (2), 436-442 (2015).</li><li>Bharat, A., Jaenicke, M., Shenoy, S. <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+technique+of+vascular+anastomosis+to+prevent+juxta-anastomotic+stenosis+following+arteriovenous+fistula+creation.">A novel technique of vascular anastomosis to prevent juxta-anastomotic stenosis following arteriovenous fistula creation.</a> Journal of Vascular Surgery. 55 (1), 274-280 (2012).</li><li>H&#246;rer, T. 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=No-touch+technique+for+radiocephalic+arteriovenous+fistula--Surgical+technique+and+preliminary+results.">No-touch technique for radiocephalic arteriovenous fistula--Surgical technique and preliminary results.</a> The Journal of Vascular Access. 17 (1), 6-12 (2016).</li><li>Sadaghianloo, N., 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=Salvage+of+early-failing+radiocephalic+fistulae+with+techniques+that+minimize+venous+dissection.">Salvage of early-failing radiocephalic fistulae with techniques that minimize venous dissection.</a> Annals of Vascular Surgery. 29 (7), 1475-1479 (2015).</li><li>Sadaghianloo, N., 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=Radial+artery+deviation+and+reimplantation+inhibits+venous+juxta-anastomotic+stenosis+and+increases+primary+patency+of+radial-cephalic+fistulas+for+hemodialysis.">Radial artery deviation and reimplantation inhibits venous juxta-anastomotic stenosis and increases primary patency of radial-cephalic fistulas for hemodialysis.</a> Journal of Vascular Surgery. 64 (3), 698-706 (2016).</li><li>Bai, H., 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=Artery+to+vein+configuration+of+arteriovenous+fistula+improves+hemodynamics+to+increase+maturation+and+patency.">Artery to vein configuration of arteriovenous fistula improves hemodynamics to increase maturation and patency.</a> Science Translational Medicine. 12 (557), (2020).</li><li>Zhang, Y. Y., Wang, X. H., Liu, Z., Hou, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Creating+radio-cephalic+arteriovenous+fistula+in+the+forearm+with+a+modified+no-touch+technique.">Creating radio-cephalic arteriovenous fistula in the forearm with a modified no-touch technique.</a> Journal of Visualized Experiments. (182), e62784 (2022).</li><li>Hou, G. C., 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=Modified+no-touch+technique+for+radio-cephalic+arteriovenous+fistula+increases+primary+patency+and+decreases+juxta-anastomotic+stenosis.">Modified no-touch technique for radio-cephalic arteriovenous fistula increases primary patency and decreases juxta-anastomotic stenosis.</a> The Journal of Vascular Access. , (2022).</li><li>Kunlin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+vein+transplantation+in+treatment+of+ischemia+caused+by+arteritis.">Long vein transplantation in treatment of ischemia caused by arteritis.</a> Revue de Chirurgie. 70 (7-8), 206-235 (1951).</li><li>Wang, Y., 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=Venous+stenosis+in+a+pig+arteriovenous+fistula+model--Anatomy,+mechanisms+and+cellular+phenotypes.">Venous stenosis in a pig arteriovenous fistula model--Anatomy, mechanisms and cellular phenotypes.</a> Nephrology, Dialysis, Transplantation. 23 (2), 525-533 (2008).</li><li>Marius, C. 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=Sheep+model+of+hemodialysis+arteriovenous+fistula+using+superficial+veins.">Sheep model of hemodialysis arteriovenous fistula using superficial veins.</a> Seminars in Dialysis. 28 (6), 687-691 (2015).</li><li>Ramacciotti, E., 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=Fistula+size+and+hemodynamics:+An+experimental+model+in+canine+femoral+arteriovenous+fistulas.">Fistula size and hemodynamics: An experimental model in canine femoral arteriovenous fistulas.</a> The Journal of Vascular Access. 8 (1), 33-43 (2008).</li><li>Eiketsu, 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=Arterial+enlargement,+tortuosity,+and+intimal+thickening+in+response+to+sequential+exposure+to+high+and+low+wall+shear+stress.">Arterial enlargement, tortuosity, and intimal thickening in response to sequential exposure to high and low wall shear stress.</a> Journal of Vascular Surgery. 39 (3), 601-612 (2004).</li><li>Eddie, 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=A+new+arteriovenous+fistula+model+to+study+the+development+of+neointimal+hyperplasia.">A new arteriovenous fistula model to study the development of neointimal hyperplasia.</a> Journal of Vascular Research. 49 (2), 123-131 (2012).</li><li>Karl, A. N., 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+murine+dialysis+fistula+model+exhibits+a+senescence+phenotype:+pathobiological+mechanisms+and+therapeutic+potential.">The murine dialysis fistula model exhibits a senescence phenotype: pathobiological mechanisms and therapeutic potential.</a> American Journal of Physiology. Renal Physiology. 315 (5), 1493-1499 (2018).</li><li>Hong, S. Y., 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=Clinical+analysis+of+radiocephalic+fistula+using+side-to-side+anastomosis+with+distal+cephalic+vein+ligation.">Clinical analysis of radiocephalic fistula using side-to-side anastomosis with distal cephalic vein ligation.</a> The Korean Journal of Thoracic and Cardiovascular Surgery. 46 (6), 439-443 (2013).</li><li>Tang, W. G., 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+meta-analysis+of+traditional+and+functional+end-to-side+anastomosis+in+radiocephalic+fistula+for+dialysis+access.">A meta-analysis of traditional and functional end-to-side anastomosis in radiocephalic fistula for dialysis access.</a> International Urology and Nephrology. 53 (7), 1373-1382 (2021).</li><li>Marie, Y., 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=Patterns+of+blood+flow+as+a+predictor+of+maturation+of+arteriovenous+fistula+for+haemodialysis.">Patterns of blood flow as a predictor of maturation of arteriovenous fistula for haemodialysis.</a> The Journal of Vascular Access. 15 (3), 169-174 (2014).</li><li>Srivastava, A., 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=Spiral+laminar+flow,+the+earliest+predictor+for+maturation+of+arteriovenous+fistula+for+hemodialysis+access.">Spiral laminar flow, the earliest predictor for maturation of arteriovenous fistula for hemodialysis access.</a> Indian Journal of Urology. 31 (3), 240-244 (2015).</li><li>Loveland-Jones, C. E., 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+new+model+of+arteriovenous+fistula+to+study+hemodialysis+access+complications.">A new model of arteriovenous fistula to study hemodialysis access complications.</a> The Journal of Vascular Access. 15 (5), 351-357 (2014).</li><li>Wong, C. Y., 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=Vascular+remodeling+and+intimal+hyperplasia+in+a+novel+murine+model+of+arteriovenous+fistula+failure.">Vascular remodeling and intimal hyperplasia in a novel murine model of arteriovenous fistula failure.</a> Journal of Vascular Surgery. 59 (1), 192-201 (2014).</li></ol>

Currently, several animal models are available for AVF. Among them, pigs, sheep, and dogs are mostly used as large animal models13,14,15. The small animal models used include rabbits, rats, and mice16,17,18. New Zealand rabbits were used in this study. New Zealand rabbits have abundant perivenous tissues around the EJV, which makes them...

The construction of an arteriovenous fistula (AVF) is widely used in clinical practice for patients undergoing maintenance hemodialysis (MHD), and it has higher patency and fewer complications than an arteriovenous graft (AVG) or tunneled cuffed catheter (TCC)1,2. Although AVF is the preferred mode of vascular access, it is not perfect and has inherent limitations. The 1 year primary AVF patency rates are only 60%-65%, with many failures occurring in the near anastomotic region3,4,5.
Vessels undergo different degrees of damage during the traditional surgical approach, which ultimately affects the maturation of the AVF. New surgical modalities, such as the no-touch technique (NTT)&#160;(Supplementary Figure 1) proposed by H&#246;rer et al.6 and radial artery excursion and reimplantation (RADAR) proposed by Sadaghianloo et al.7,8 and Bai et al.9, were designed to decrease the rate of juxta-anastomotic stenosis and to improve the fistula patency by modifying the surgical technique. Although the effect of RADAR was better than that of NTT, inflow arterial stenosis was observed to be more prominent with RADAR. To further reduce injury to the veins and arteries during the operation, in 2021, a new modified no-touch technique (MNTT) was proposed to create a radio-cephalic AVF by preserving the perivenous tissue around the cephalic vein without cutting the radial artery (Supplementary Figure 1 and Supplementary Figure 2). The preliminary results showed increased primary patency, decreased juxta-anastomotic stenosis, and no arterial stenosis10,11.
Considering the current lack of animal models of AVF using MNTT, and to further explore the mechanism of MNTT in AVF surgery, this study introduces a common carotid artery (CCA)-external jugular vein (EJV) AVF procedure using MNTT.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Animal Depilatory</td>
			<td>Fuzhou Feijing Biotechnology Co., Ltd.</td>
			<td>PH1877</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved hemostatic forceps</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZH131R/RN</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting forceps</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZDO25R/RN</td>
			<td></td>
		</tr>
		<tr>
			<td>electrical razor</td>
			<td>Shenbao Technology Co., Ltd</td>
			<td>PGC-660</td>
			<td></td>
		</tr>
		<tr>
			<td>Fixed Table</td>
			<td>Zhenhua Biomedical Instrument Co., Ltd</td>
			<td>ZH-DSB019</td>
			<td></td>
		</tr>
		<tr>
			<td>Halsey needleholder</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZM208R/RN</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin Dodium Injection</td>
			<td>Jiangsu Wanbang Biochemical Pharmaceutical Group Co., Ltd.</td>
			<td>H32020612</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical gauze dressing</td>
			<td>Nanchang Kangjie medical hygiene products Co., Ltd</td>
			<td>20172640135</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceops</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZD275RN/T</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro needle holder forceps</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZF2618RB/T</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZF022T</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-silk sutures 4-0</td>
			<td>Kollsut Medical Instrument Co., Ltd.</td>
			<td>NMB020RRCN26C075-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-absorbable sutures 8-0 (double needle)</td>
			<td>Yangzhou Yuankang Medical Instrument Co., Ltd.</td>
			<td>10299023602</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone iodine solution</td>
			<td>Shanghai Likang Disinfection High-tech Co., Ltd.</td>
			<td>310512</td>
			<td></td>
		</tr>
		<tr>
			<td>Rinse needle</td>
			<td>Jiangsu Tonghui Medical Instrument Co., Ltd</td>
			<td>20180039</td>
			<td></td>
		</tr>
		<tr>
			<td>scalpel handle</td>
			<td>Shanghai Medical Instrument (Group) Co., Ltd. Surgical Instruments Factory</td>
			<td>J11030</td>
			<td></td>
		</tr>
		<tr>
			<td>Sharp blade</td>
			<td>Suzhou Medical Products Factory Co., Ltd.</td>
			<td>TY21232001</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride Injection&#160; (100 mL)</td>
			<td>Guangdong Otsuka Pharmaceutical Co., Ltd.</td>
			<td>B21K0904</td>
			<td></td>
		</tr>
		<tr>
			<td>Sugical Scissors</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZC120R/RN</td>
			<td></td>
		</tr>
		<tr>
			<td>Sumianxin II</td>
			<td>Jilin Dunhua Shengda Animal Pharmaceutical Co., Ltd.</td>
			<td>20180801</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe with needle&#65288;5 mL)</td>
			<td>BD medical devices (Shanghai) Co., Ltd</td>
			<td>2006116</td>
			<td></td>
		</tr>
		<tr>
			<td>Tiletamine Hydrochloride and Zolazepam Hydrochloride for Injection</td>
			<td>Virbac Pet Health, France</td>
			<td>83888204</td>
			<td></td>
		</tr>
		<tr>
			<td>Triangle needle</td>
			<td>Hangzhou Huawei medical supplies Co., Ltd</td>
			<td>7X17</td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular clamp</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZF220RN</td>
			<td></td>
		</tr>
		<tr>
			<td>New Zealand rabbits</td>
			<td>Suzhou Huqiao Biological Co., Ltd.</td>
			<td>SCXK2020-0001</td>
			<td></td>
		</tr>
	</tbody>
</table>

a modified technique for arteriovenous fistula construction in rabbits

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury to the veins and arteries during the operation and hemodynamic changes can lead to intimal hyperplasia, leading to juxta-anastomotic stenosis. To reduce injury to the veins and arteries during the operation, this study proposes a new modified no-touch technique (MNTT) for AVF construction that can decrease the rate of juxta-anastomotic stenosis and improve the AVF patency. To unravel the hemodynamic changes and mechanisms of the MNTT, this study presented an AVF procedure using this technique. Although this procedure is technically challenging, 94.4% procedural success was achieved after adequate training. Ultimately, 13 out of 34 rabbits had a functional AVF 4 weeks after the surgery, leading to a 38.2% AVF patency rate. However, at 4 weeks, the survival rate was 86.1%. Ultrasonography showed active blood flow through AVF anastomosis. Furthermore, the spiral laminar flow was observed in the vein and artery near the anastomosis, suggesting that this technique may improve the hemodynamics of the AVF. On histological observation, significant venous intimal hyperplasia was observed at the AVF anastomosis, whereas no significant intimal hyperplasia was observed at the proximal external jugular vein (EJV) of the anastomosis. This technique will improve the understanding of the mechanisms underlying the use of MNTT for AVF construction and provide technical support for the further optimization of the surgical approach in AVF construction.

The outcome of the successful application of this technique is a patent AVF in the rabbit neck. This study used the following criteria to evaluate the success: (1) when the vascular anastomosis is completed, the venous tremor of the AVF can be touched, and the vascular murmur can be heard; (2) 4 weeks after the AVF is established, the active blood flow through the internal fistula anastomosis can be measured by color Doppler ultrasound; (3) 4 weeks after the AVF is established, hematoxylin-eosin (H&#38;E) staining shows ...

Watch this Scientific Journal Video about A Modified Technique for Arteriovenous Fistula Construction in Rabbits at JoVE.com

A Modified Technique for Arteriovenous Fistula Construction in Rabbits

The present protocol proposes the creation of an arteriovenous fistula in rabbits using a modified no-touch technique. The technique involves the side-to-side anastomosis of the common carotid artery and external jugular vein without the dissection of the perivenous tissues or cutting off the artery.

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury ...

Clinically, radiocephalic AVF created with a modified no-touch technique can increase primary patency and decrease juxta-anastomotic stenosis. To explore the mechanism of MNTT, this animal model needs to be established. The main advantage of the modified no-touch technique is to reduce injury to the external jugular vein when establishing AVF.When dealing with the external jugular vein care should be taken to preserve the paravenous tissue as much as possible and reduce the injury to the vein. To begin, place an anesthetized rabbit in a supine position on a fixed table. Bind its limbs and incisors.Using an electric razor, shave the rabbit's neck and upper chest, and remove the hair with animal depilatory cream. Next, using a scissor, make a three centimeter long incision between the mandible and the sternoclavicular joint. Identify the right extra jugular vein, or EJV, and make a tunnel perpendicular to it.Place a vascular clamp along the tunnel and similarly make another tunnel at the distal EJV. Control the blood flow by applying a 4-O suture and a clamp along the tunnel. To dissect the common carotid artery, begin by exploring the CCA lateral to the trachea and medial to the sternocleidomastoid muscle.Now, dissect two centimeters of the CCA. Using a 4-0 suture thread, control the blood flow when needed. Place vascular clamps at the distal and proximal lens of the CCA.For phlebotomy and anastomosis, use micro scissors to dissect four millimeters of the internal part of the EJV. Then, make a four millimeter long incision in the middle of the vein. Rinse the vein with heparin solution to prevent thrombosis.Make a similar incision in the anterior wall of the artery and rinse as demonstrated. Pull the EJV and the CCA as close together as possible. Apply Connell's technique using 8-0 non-absorbable sutures to perform side-by-side anastomosis of the vessels.Suture the posterior wall of the vessels first, followed by the anterior vessel wall. To observe active blood flow through the anastomosis, remove all vascular clamps except for the one at the distal EJV. Using the 4-0 suture placed earlier, ligate the distal end of the EJV, then remove the distal clamp.Remove the suture thread around the CCA. Of the 34 rabbits that had immediately successful arterial venous fistula using the modified no-touch technique, only 31 rabbits survived, resulting in a survival rate of 86.1%13 out of 34 rabbits had a functional AVF four weeks after the surgery leading to a 38.2%AVF patency rate. The patent arterial venous fistula was confirmed after four weeks.Spiral laminar flow was observed in both vessels near the anastomosis. Upon HE staining, significant venous intimal hyperplasia was observed at the site of the arteriovenous fistula anastomosis, but not at the proximal EJV of the anastomosis. AVF animal model established using MNTT can provide technical support for further exploration of the molecular biological mechanism of juxta-anastomotic stenosis.

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