We are supported by research grants from the National Natural Science Foundation of China (NO. 81400590), National Institutes of Health (U54 CA156735) and Takeda Pharmaceutical Company Ltd. (MA-NC-D-156).

모든 동물 실험 기관 동물 케어 및 사용위원회에 의해 승인되었습니다. 1. 마우스 준비 <ol><li> 을 통해 마우스를 사용합니다 ~ 체중 20g 또는 수술 연령 6 ~ 위의 주. </li><li> 수술 전, 쥐에게 실험 차우와 물 광고 무제한을 제공하고 12시 12분 시간의 명암주기를 유지한다. </li><li> 머리 깎기와 수술 영역을 면도. 80 밀리그램 / kg 케타민, 12 밀리그램 / kg 자일 라진의 복강 내 주사하여 마우스를 마취. 이 용량은 몇 분 안에 잠에 마우스를두고 다음과 같은 외과 적 치료에 대한 충분한 마취를 제공합니다. </li><li> 발바닥이 끼 때 각막 반사와 사지 수축의 부재에 따라 적절한 마취를 확인합니다. 건조를 방지하기 위해 눈에 안과 연고를 윤활 적용합니다. </li><li> betadine 용액으로 피부를 소독. </li></ol> 2. 위 역류 모델 (그림 1B) <ol><li> 위 확인항문을 가리키는 검상 돌기에서 시작 ~ 2cm의 중간 선 절개. </li><li> 중간 선을 통해 복강을 엽니 다. 가위와 노출을 향상시키기 위해 칼 모양을 제거합니다. </li><li> 별도의 간 및 위장 사이의 결합 조직을 잘라. 결찰 및 주문 안저까지 완전 무료로 비장과 안저의 혈관 번들을 잘라. </li><li> 왼쪽으로 약간 안저를 켜고 분야에서 위식도 접합부의 왼쪽에 노출됩니다. 상피를 노출 예리한 가위를 사용하여 식도 따라 근육에 길이 5mm 절개를. </li><li> 같은 방향 (절개 1, 그림 1B)를 따라 상피를 오픈. (절개 2, 3, 4, 그림 1B) 날카로운 운영 가위 forestomach의 대부분을 차단하기 위해 forestomach에 세 개의 절개를합니다. 위가 비어 있지 않은 경우,주의 깊게 위 내용물을 제거합니다. </li><li> 8-0 prolene의의 수 투어를 배치재 정확한 점막에 지점을 통해 (forestomach에) (식도에) 및 포인트 &#39;(테이퍼 포인트 바늘) 반대 점막합니다. 각각 점 B와 점 B &#39;, 그리고 포인트의 다른 쌍을 통해 마찬가지로 장소 봉합. 균등 공간이 봉합 서로 3-4mm. </li><li> 혈액과 위의 내용을 정리하는 생리 식염수로 복강을 세척 할 것. 실크 봉합사와 복부 벽과 금속 클립으로 피부를 닫습니다. </li></ol> 3. 혼합 환류 모델 (그림 1C) <ol><li> 팔로우는 위식도 접합부를 노출 2.4 2.1 단계를 반복합니다. </li><li> 부드럽게 식도 뒤에 혈관에서 식도의 등 쪽을 분리합니다. 식도과 혈관 사이의 작은면 팁을 전달합니다. 주 :면이 부분적으로 그 크기를 줄이기 위해 선단으로부터 제거 될 수있다. 이면 팁 식도를 해제하고 혈관을 보호 두 가지 목적. </li><li> 두 5mm 길이 INC 확인위식도 접합부와 날카로운 운영 가위로 유문에 인접한 십이지장의 기단부에 isions 각. 십이지장에 절개를 들어, 혈관을 피하고 항 장간막 테두리에 배치했다. </li><li> 중단 8-0 prolene의 봉합과 반대 점막 정확한 점막에 절개를 문합. 일반적으로 등쪽에 3-4 봉합과 전면에 2-3 봉합을 배치합니다. </li><li> 면 팁을 제거합니다. 생리 식염수와 복강을 세척하고 복벽과 피부를 닫습니다. </li></ol> 4. 십이지장 역류 모델 (그림 1D) <ol><li> 혼합 역류를 생성하기 위해 3.4에 3.1 단계를 따르십시오. </li><li> 조심스럽게 뒷면을 노출 위장을 들어 올립니다. 간 로브는 간, 위의 배면 사이의 결합 조직에 의해 잡힐 수있다. 결합 조직을 조심스럽게 잘라 적절히 간을 보호하고, 식도의 등쪽에 혈관을 노출. </li><li> 결찰 및혈관을 절단. 결찰 및 위식도 접합부에서 식도를 잘라. </li><li> 유문에 십이지장을 결찰. 결찰 및 mesenterium를 잘라. 전체 위장을 제거합니다. </li><li> 생리 식염수로 복강을 세척 할 것. 복벽과 피부를 닫습니다. </li></ol> 5. 후 수술 적 치료 <ol><li> 수술 후 가열 패드와 체온을 유지한다. 만 흉골 드러 누움을 회복 한 후, 다시 다른 동물과 마우스를 놓습니다. </li><li> 감염을 예방하고 통증을 완화하기 위해 항생제와 진통제를 준다. Baytril를 주입 (10 ㎎ / ㎏, IP, QD, 3 일간) 감염을 방지하고, 부 프레 노르 핀 염산염 (0.05 ㎎ / ㎏, SC, 입찰. 이틀 동안) 진통제로. 금식은 수술 후 필요는 없습니다. 액체 또는 부드러운식이 요법을들 수있다. </li><li> 마우스는 다음과 같은 징후를 보여 매일 basis.If에 일반 건강 상태 (극적인 체중 감소&gt; 15 %, 응용 프로그램의 부족을 평가etite, 발성, 쥐를 안락사, 입, 코, 눈, 구부리고 제스처, 활동, 비정상적인 정리 활동)에서 배출. </li></ol>

<ol>
	<li>Kandulski, A., Malfertheiner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroesophageal+reflux+disease--from+reflux+episodes+to+mucosal+inflammation.">Gastroesophageal reflux disease--from reflux episodes to mucosal inflammation.</a> Nat Rev Gastroenterol Hepatol. 9 (1), 15-22 (2012).</li><li>Orlando, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+integrity+of+the+esophageal+mucosa.+Balance+between+offensive+and+defensive+mechanisms.">The integrity of the esophageal mucosa. Balance between offensive and defensive mechanisms.</a> Best Pract Res Clin Gastroenterol. 24 (6), 873-882 (2010).</li><li>Stairs, D. B., Kong, J., Lynch, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cdx+genes,+inflammation,+and+the+pathogenesis+of+intestinal+metaplasia.">Cdx genes, inflammation, and the pathogenesis of intestinal metaplasia.</a> Prog Mol Biol Transl Sci. 96, 231-270 (2010).</li><li>Spechler, S. J., Fitzgerald, R. C., Prasad, G. A., History Wang, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=molecular+mechanisms,+and+endoscopic+treatment+of+Barrett's+esophagus.">molecular mechanisms, and endoscopic treatment of Barrett's esophagus.</a> Gastroenterology. 138 (3), 854-869 (2010).</li><li>Fang, 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=Cellular+origins+and+molecular+mechanisms+of+Barrett's+esophagus+and+esophageal+adenocarcinoma.">Cellular origins and molecular mechanisms of Barrett's esophagus and esophageal adenocarcinoma.</a> Ann N Y Acad Sci. 1300, 187-199 (2013).</li><li>Garman, K. S., Orlando, R. C., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review:+Experimental+models+for+Barrett's+esophagus+and+esophageal+adenocarcinoma.">Review: Experimental models for Barrett's esophagus and esophageal adenocarcinoma.</a> Am J Physiol Gastrointest Liver Physiol. 302 (11), G1231-G1243 (2012).</li><li>Baruah, 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=Translational+research+on+Barrett's+esophagus.">Translational research on Barrett's esophagus.</a> Ann N Y Acad Sci. 1325, 170-186 (2014).</li><li>Lechpammer, 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=Flavopiridol+reduces+malignant+transformation+of+the+esophageal+mucosa+in+p27+knockout+mice.">Flavopiridol reduces malignant transformation of the esophageal mucosa in p27 knockout mice.</a> Oncogene. 24 (10), 1683-1688 (2005).</li><li>Hao, J., Liu, B., Yang, C. S., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroesophageal+reflux+leads+to+esophageal+cancer+in+a+surgical+model+with+mice.">Gastroesophageal reflux leads to esophageal cancer in a surgical model with mice.</a> BMC Gastroenterol. 9, 59 (2009).</li><li>Mari, 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+pSMAD/CDX2+complex+is+essential+for+the+intestinalization+of+epithelial+metaplasia.">A pSMAD/CDX2 complex is essential for the intestinalization of epithelial metaplasia.</a> Cell Rep. 7 (4), 1197-1210 (2014).</li><li>Fang, 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=Gastroesophageal+reflux+activates+the+NF-kappaB+pathway+and+impairs+esophageal+barrier+function+in+mice.">Gastroesophageal reflux activates the NF-kappaB pathway and impairs esophageal barrier function in mice.</a> Am J Physiol Gastrointest Liver Physiol. 305 (1), G58-G65 (2013).</li><li>Chen, 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=Nrf2+deficiency+impairs+the+barrier+function+of+mouse+oesophageal+epithelium.">Nrf2 deficiency impairs the barrier function of mouse oesophageal epithelium.</a> Gut. 63 (5), 711-719 (2014).</li></ol>

One of the authors (XC) received funding that was provided by Takeda Pharmaceutical Company Ltd which supports basic research associated with gastroesophageal reflux disease. None of the reagents or instruments used in this article is produced by this company.

다양한 수술 모델은 설치류에서 위, 십이지장 및 혼합 역류을 모방하기 위해 설립되었다. 여기에 설명이 세 가지 절차는 수술 후 생존의 합리적인 가격과 장기적인 실험에 적합하다. 수술 경험이있는 연구원은 연습의 짧은 기간 후에 기술을 파악 할 수 있어야한다. 출혈은 수술, 간, 위, 및 혈관의 부주의 한 손상 사이의 결합 조직의 분리시 간 열상 전 마취제의 복강 내 주사에 기인 할 수있다. 이것은 가능한 한 피해야한다. 배터리 구동 미니 소작 적절한 경우 지혈하는데 사용될 수있다. 기관 및 조직이 조작 될 때 과도한 극력 피해야 연신. 핀셋의 끝은면으로 포장 될 수있다, 또는면 팁 핀셋을 대체하기 위해 젓가락처럼 사용할 수있다. 마우스 식도는 두 개의 층 또는 두 개의 &amp; #이8220; 관 &quot;, 외부 붉은&quot;근육 관 &quot;과 내부 희끄무레&quot;상피 관 &quot;. 절개는 식도에 이루어질 때, 양 층은 절개 할 필요가있다. 봉합은 균등하게 반대 점막 정확한 점막과 간격을해야합니다. 협착이 봉합 사이의 좁은 거리로 인해 생길 수있는 반면 누설, 봉합 사이의 넓은 거리에서 발생할 수 있습니다. 그러나, 협착은 음식의 흐름을 차단할 수 있습니다 약간의 누설이 지역의 접착과 섬유화 반응에 의해 고정 될 수있는 반면, 수술 후 사망에이를 수 있으므로 일반적으로 생명을 위협하지 않습니다. 프로토콜 내에서 따라야 할 몇 가지 비판적 일반 원칙이있다. (실험실에서 약 20-30 분)이 가능하면 수술 시간을 단축해야한다. 수술 전후 금식하는 것은 불필요하다. 사실, 장기간 금식 (&gt; 일일)은 유해 할 수 있습니다. 수술은 가장 한 외과 의사에 의해 수행된다. 조수는 쥐를 마취하고 surg를 제공 도움이 될 수 있습니다iCal을 재료. 외과 의사가 제대로 미세 훈련되지 않는 한 현미경이 필요하지 않습니다. 달리 수술 기간 이상 필요하므로 수술 생존 가능성 악화 될 수보다 쉽다. 이러한 절차는 이전의 공보에서 사용되어왔다과 만족스러운 결과를 생성 하였다. 야생 형 및 유전자 변형 쥐를 사용하여, 우리는 NFκB와 Nrf2 경로는 위식도 역류 (11, 12) 중 식도 상피의 장벽 기능을 조절 것을 보여 주었다. 그러나, 대신에, 이들 마우스에서 본격적인 바렛 식도 선암종 또는 생산할 수 없었다 점액 세포와 편평 상피 세포 암 (9)을 유도 하였다 흩어져. 마우스 식도 및 인간의 식도 사이의 조직 학적 차이는 바렛 식도 및 식도 선암에 대한 연구에서 이러한 수술 모델의 고유 제한 될 수 있습니다. 우리는이 SU를 생각rgical 모델은 위 식도 역류 질환에 대한 연구에 유용하다. 유전자 및 분자 경로의 기능 GERD의 분자 메커니즘을 이해하고기구 기반 치료법​​을 개발하기 위해 설명 될 수있다. 바렛 식도 및 식도 선암종의 제조에 사용하는 경우, 유전자 변형은 수술 환류와 함께 필요하다.

Gastroesophageal reflux disease (GERD) is a chronic disorder caused by the prolonged exposure of distal esophagus to gastric or gastroduodenal contents1. Prolonged exposure to these noxious refluxates impairs the intrinsic defenses within the esophageal epithelium and thus results in esophagitis2. Barrett&#8217;s esophagus arises in the setting of chronic reflux, and is a premalignant lesion with increased risk of esophageal adenocarcinoma3,4. Despite the clinical importance, the mechanisms of GERD, Barrett&#8217;s esophagus and adenocarcinoma have not been well understood. 
Animal models are essential for research on etiology, pathology, molecular mechanisms, prevention and treatment of human diseases. Up to date, various animal models of GERD, Barrett&#8217;s esophagus and adenocarcinoma have been developed using model animals5,6. Mouse esophagus is lined with stratified squamous epithelium which is histologically similar to that in human esophagus. Although a mouse esophagus is different from human esophagus in terms of keratinization and the absence of submucosal glands, the mouse is still an appealing model animal because of its relatively low cost of maintenance and its potential of sophisticated genetic modifications. Two approaches are commonly used to model GERD, Barrett&#8217;s esophagus and adenocarcinoma in mice: reflux surgery and genetic modification. Reflux surgery is the best way to induce reflux and genetic modifications mimics molecular alterations5,7. Reflux surgery can be combined with genetic modifications to further understand disease mechanisms8. 
Many surgical procedures have been reported by us and others6,9: (1) gastric reflux: pyloric ligation, pyloric constriction with forestomach ligation, Wendel cardioplasty, and esophagogastric anastomosis; (2) mixed reflux: esophagogastroduodenal anastomosis, esophagoduodenostomy (or esophagojejunostomy); (3) duodenal reflux: esophagogastroduodenal anastomosis plus gastrectomy; (4) reflux of chemical components: bilious reflux, pancreatic reflux, esophageal perfusion; and (5) esophageal transplantation5. Recently a microsurgical mouse model was reported to produce jejunal reflux via an esophagojejunostomy with magnets10. These surgical models have advantages over in vitro cell culture or organotypic culture models. In vitro, esophageal cells cannot tolerate a medium with high acidity or high concentrations of bile acids. Unconjugated bile acids which are commonly used to produce changes in esophageal epithelial cells in vitro are usually not present in the duodenal refluxate in vivo. Thus conclusions drawn from such in vitro studies should be taken with caution.
Surgery on the mouse esophagus remains a technical challenge because of its small size. A low rate of postoperative survival does not allow experiments which require certain sample size to reach statistically sound conclusions. In the past we have successfully developed and characterized surgical models of gastric reflux, mixed reflux, duodenal reflux with mice in long-term experiments9,11,12. We have also provided consultation to several other groups in their mouse surgery. Herein, we describe three surgical procedures in mice in order to help the community to establish these models in their labs.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Instruments</td>
			<td>Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #1 Forceps Dumostar Tip&#160;</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Clip Applying Forceps 5.5&#34;</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bonn Scissors 3.5&#34; Straight 15mm Sharp/Sharp Tungsten Carbide Blades</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Operating Scissors 5.5&#34; Straight Sharp/Sharp SureCut</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 Silk Black Braid 100 Yard Spool</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgeon&#39;s Needle 1/2 Circle Cutting Edge Size 12 (25 mm Chord Length) Pack 12</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Halsey Needle Holder 5&#34; Smooth</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Needle Holder 5.125&#34; Curved Lock .6mm</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex 9mm Wound Clip Applier</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex 9mm Wound Clips Box Of 100</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PRONOVA Poly (hexafluoropropylene-VDF) Suture 8-0</td>
			<td>Ethicon US, LLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reagents</td>
			<td>Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer&#39;s solution</td>
			<td>Henry Schein, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ketamine</td>
			<td>Henry Schein, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>xylazine</td>
			<td>Henry Schein, Inc.</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

surgical models of gastroesophageal reflux with mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

대부분의 마우스는 (&gt; 95 %)이 수술을 견딜 수. 수술 전후 기간 동안 사망의 주요 원인은 마취제의 과다, 출혈 및 알 수없는 이유를들 수있다. 수술 후 4 주&gt; 십이지장 역류과 위 역류 또는 혼합 역류&gt; 80 % 마우스와 90 %의 마우스는 살아남을 수 있습니다. 이 기간 동안, 마우스는 주로 식도 협착과 먹을 수없는 사망. 이 마우스는 심한 스트레스 (구부리고 자세, 활동, 구토, 침몰 한 눈, 발성 등)의 흔적을 보여 안락사 될 필요가있다. 일반적으로 항생제 투여와 의료 죽어에서이 마우스를 저장할 수 없습니다. 마우스는 일반적으로 수술 후 그램의 부부 십이지장 역류와 특히 자신의 체중을 잃게됩니다. 그럼에도 불구하고 체중은 회복 이후에 증가한다. 마흔 주 수술 후, 십이지장 역류와 위 및 혼합 역류와&gt; 70 % 마우스와&gt; 80 % 마우스는 살아남을 수 있습니다. 지속이 기간을 보내고, 일부 마우스는 영양 실조 또는 알 수없는 이유로 사망 할 수 있습니다. 우리는 수술 후 80주을 위해 마우스를 유지하고있다. 일반적으로, 그들은 여전히 매우 활동적이고 (9) 건강. 수술 후 사주에서 식도 상피의 구조는 역류에 의해 어느 정도 손상됩니다. 현미경 하에서 상피 두꺼워지고 돌기가 뻗어 형성. 상피 세포와 염증 세포 (예를 들면, 호중구, 비만 세포와 호산구)의 침투의 과다 증식 (그림 2) 식도염의 명백한 증거입니다. 식도 상피 (예를 들면, IL1β, IL6 및 IL8)의 사이토 카인, 특히 십이지장 환류 (11)의 존재하에 증가된다. 투과 전자 현미경, 팽창 상피 세포 간 공간은 장벽 (12)의 기능 장애를 반영한다. 기능 - 방식, 트랜스 - 상피 전기 저항은 특히 duoden의 존재하에, 극적으로 감소알 역류. <img alt="그림 1" src="/files/ftp_upload/53012/53012fig1.jpg" /> 그림 1 : (A) 마우스 (정면 및 측면보기)에서 상부 소화 기관의 해부학; (B) 위 역류 (측면도) (C)를 혼합 환류 (전면보기) (D) 십이지장 역류 (전면보기). <img alt="그림 2" src="/files/ftp_upload/53012/53012fig2.jpg" /> 도 2 : 마우스 식도 상피의 조직학 환류 (A)없이, 또는 위산 역류 (B), 혼합 된 환류 (C), 십이지장 환류 (D)와. 스케일 바는 50 μm의 =.

Watch this Scientific Journal Video about Surgical Models of Gastroesophageal Reflux with Mice at JoVE.com

Surgical Models of Gastroesophageal Reflux with Mice

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

마우스와 위식도 역류의 외과 모델

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

surgical-models-of-gastroesophageal-reflux-with-mice

Universidad San Sebastian

Research

JoVE Journal

Medicine

10.0K 조회수.  Ningxia Medical University General Hospital. This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett’s esophagus and esophageal adenocarcinoma.

비디오: Surgical Models of Gastroesophageal Reflux with Mice

Surgical Management of Meatal Stenosis with Meatoplasty

Meatal stenosis is a common urologic complication after circumcision. Children present to their primary care physicians with complaints of deviated urinary stream, difficult-to-aim, painful urination, and urinary frequency. Clinical exam reveals a pinpoint meatus and if the child is asked to urinate, he will usually have an upward, thin, occasionally forceful urinary stream with incomplete bladder emptying. The mainstay of management is meatoplasty (reconstruction of the distal urethra /meatus). This educational video will demonstrate how this is performed.

Meatoplasty, surgical management of meatal stenosis.

Meatoplasty과 Meatal 협착증의 수술 관리

Meatal stenosis is a common urologic complication after circumcision.  Children present to their primary care physicians with complaints of deviated ...

연구

의학

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a manageable size and relatively easy to house. These factors, together with the availability of a wealth of spontaneous and experimentally induced mutants, make laboratory mice ideally suited to a wide variety of research areas.
In cardiovascular, pharmacological and toxicological research, accurate measurement of parameters relating to the circulatory system of laboratory animals is often required. Determination of heart rate, heart rate variability, and duration of PQ and QT intervals are based on electrocardiogram (ECG) recordings. However, obtaining reliable ECG curves as well as physiological data such as core body temperature in mice can be difficult using conventional measurement techniques, which require connecting sensors and lead wires to a restrained, tethered, or even anaesthetized animal. Data obtained in this fashion must be interpreted with caution, as it is well known that restraining and anesthesia can have a major artifactual influence on physiological parameters1, 2.
Radiotelemetry enables data to be collected from conscious and untethered animals. Measurements can be conducted even in freely moving animals, and without requiring the investigator to be in the proximity of the animal. Thus, known sources of artifacts are avoided, and accurate and reliable measurements are assured. This methodology also reduces interanimal variability, thus reducing the number of animals used, rendering this technology the most humane method of monitoring physiological parameters in laboratory animals3, 4. Constant advancements in data acquisition technology and implant miniaturization mean that it is now possible to record physiological parameters and locomotor activity continuously and in realtime over longer periods such as hours, days or even weeks3, 5.
Here, we describe a surgical technique for implantation of a commercially available telemetry transmitter used for continuous measurements of core body temperature, locomotor activity and biopotential (i.e. onelead ECG), from which heart rate, heart rate variability, and PQ and QT intervals can be established in freeroaming, untethered mice. We also present pre-operative procedures and protocols for post-operative intensive care and pain treatment that improve recovery, well-being and survival rates in implanted mice5, 6.

A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.

ECG, 심장 박동, 코어 바디 온도 및 자유 이동 실험실 마우스의 활동에 데이터를 항복 Radiotelemetry 송신기의 주입

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

Protocols for Assessing Radiofrequency Interactions with Gold Nanoparticles and Biological Systems for Non-invasive Hyperthermia Cancer Therapy

Cancer therapies which are less toxic and invasive than their existing counterparts are highly desirable. The use of RF electric-fields that penetrate deep into the body, causing minimal toxicity, are currently being studied as a viable means of non-invasive cancer therapy. It is envisioned that the interactions of RF energy with internalized nanoparticles (NPs) can liberate heat which can then cause overheating (hyperthermia) of the cell, ultimately ending in cell necrosis.
In the case of non-biological systems, we present detailed protocols relating to quantifying the heat liberated by highly-concentrated NP colloids. For biological systems, in the case of in vitro experiments, we describe the techniques and conditions which must be adhered to in order to effectively expose cancer cells to RF energy without bulk media heating artifacts significantly obscuring the data. Finally, we give a detailed methodology for in vivo mouse models with ectopic hepatic cancer tumors.

We describe the protocols used to investigate the interactions of 13.56 MHz radiofrequency (RF) electric-fields with gold nanoparticle colloids in both non-biological and biological systems (in vitro/vivo). These interactions are being investigated for applications in cancer therapy.

비 침습 온열 암 치료를위한 금 나노 입자 및 생물 시스템과 고주파의 상호 작용을 평가하기위한 프로토콜

Cancer therapies  which are less toxic and invasive than their existing counterparts are highly  desirable. The use of RF electric-fields that penetrate ...

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque formation and stenosis, as observed for example in ischemic heart disease. Two highly reproducible mouse models&#160;&#8211;&#160;both resulting in artery stenosis but each underlying a different pathway of development&#160;&#8211; are introduced here. The models represent the two most common causes of artery stenosis; namely one mouse model for each myointimal hyperplasia, and atherosclerosis are shown. To induce myointimal hyperplasia, a balloon catheter injury of the abdominal aorta is performed. For the development of atherosclerotic plaque, the ApoE -/- mouse model in combination with western fatty diet is used. Different model-adapted options for the measurement and evaluation of the results are named and described in this manuscript. The introduction and comparison of these two models provides information for scientists to choose the appropriate artery stenosis model in accordance to the scientific question asked.

This video shows two models of intimal plaque development in murine arteries and emphasizes the differences in myointimal hyperplasia and atherosclerosis.

쥐에서 동맥 경화 수스 Myointimal 증식을 유도 : 두 개의 유효한 모델의 소개

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

쥐 심근 경색을 유도 최소 침습 수술

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

깊은 뇌 자극과 Saccadometry를 사용하여 정상 및 병적 뇌 기능을 연구하는

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Cutaneous Surgical Denervation:  A Method for Testing the Requirement for Nerves in Mouse Models of Skin Disease

Cutaneous somatosensory nerves function to detect diverse stimuli that act upon the skin. In addition to their established sensory roles, recent studies have suggested that nerves may also modulate skin disorders including atopic dermatitis, psoriasis and cancer. Here, we describe protocols for testing the requirement for nerves in maintaining a cutaneous mechanosensory organ, the touch dome (TD). Specifically, we discuss methods for genetically labeling, harvesting and visualizing TDs by whole-mount staining, and for performing unilateral surgical denervation on mouse dorsal back skin. Together, these approaches can be used to directly compare TD morphology and gene expression in denervated as well as sham-operated skin from the same animal. These methods can also be readily adapted to examine the requirement for nerves in mouse models of skin pathology. Finally, the ability to repeatedly sample the skin provides an opportunity to monitor disease progression at different stages and times after initiation.

This article includes detailed protocols for genetic labeling of mouse skin, surgical denervation, skin biopsy and visualizing labeled epithelia by whole-mount &#946;-galactosidase staining. These methods can be used to test the requirement for nerves in mouse models of normal and pathological skin.

탈 신경 외과 피부 : 피부 질환의 마우스 모델에서 신경에 대한 요구를 테스트하기위한 방법

Cutaneous somatosensory nerves function to detect diverse stimuli that act upon the skin. In addition to their established sensory roles, recent studies ...

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

마우스의 중간 대뇌 동맥 폐색 및 재관류 유도 스트로크에 대한 수술 적 접근

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

Intravital 현미경 종양 관련 맥 관 구조를 사용 하 여 고급 등 유전자 변형 형광 생쥐 피부 집계 창 실

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an otherwise dynamic system, and successive phases are easily overlooked or misinterpreted. Live-cell imaging and time-lapse microscopy, in which living cells can be followed for hours or even days in a more or less continuous fashion, are therefore very informative. The protocol described here allows for the investigation of the fate of chemotherapeutic nanoparticles after the delivery of doxorubicin (dox) in living cells. Dox is an intercalating agent that must be released from its nanocarrier to become biologically active. In spite of its clinical registration for more than two decades, its uptake, breakdown, and drug release are still not fully understood. This article explores the hypothesis that lipid-based nanoparticles are taken up by the tumor cells and are slowly degraded. Released dox is then translocated to the nucleus. To prevent fixation artifacts, live-cell imaging and time-lapse microscopy, described in this experimental procedure, can be applied.

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Live-cell imaging is a powerful tool to visualize dynamic processes. The examination of fixed cells provides only static pictures, which can lead to misinterpretation and confusion about the process. This work presents a method to study uptake, drug release, and intracellular localization of liposomal nanoparticles in living cells.

사용 하 여 체 외에서 전달 Chemotherapeutics 탐험 라이브 셀 이미징 지질 기반 나노 입자에 의해

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an ...