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).

Tüm hayvan deneyleri Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanmıştır. 1. Fare Hazırlık <ol><li> Üzerinde fareler kullanın ~ vücut ağırlığı 20 g veya ameliyat yaşı 6 ~ yukarıdaki hafta. </li><li> Ameliyattan önce, fareler laboratuvar yemi ve su ad libitum beslendi ve 00:12 saat ışık-karanlık çevrimi ile korumak. </li><li> Saç kesme makinesi ile cerrahi alan tıraş. 80 mg / kg ketamin ve 12 mg / kg ksilazin intraperitonal enjeksiyonu ile fareler anestezi. Bu doz birkaç dakika içinde uykuya fareler koyar ve aşağıdaki cerrahi işlemler için yeterli anestezi sağlar. </li><li> Ayak tabanı sıkışmak zaman kornea refleksi ve uzuv retraksiyon yokluğunda dayalı doğru anesthetization onaylayın. Kuruluğunu önlemek için gözleri oftalmik merhem yağlama uygulayın. </li><li> Betadine solüsyonu ile cilt sterilize edin. </li></ol> 2. Mide cezir modeli (Şekil 1B) <ol><li> Bir üst yapanüs işaret kılıç şeklinde başlayarak ~ 2 cm orta hat insizyonu. </li><li> Orta hat boyunca karın boşluğu açın. Bir makas ile poz geliştirmek için kılıç şeklinde çıkarın. </li><li> Ayrı ve karaciğer ve mide arasında bağ dokuları kesmek. Bağlanır ve sipariş fundus kadar tamamen ücretsiz olarak dalak ve fundus arasındaki damar demeti kesti. </li><li> Sola hafifçe fundus çevirin ve bu alandaki gastroözofageal bileşke sol tarafını ortaya çıkarmak. Epitel maruz keskin bir makas kullanarak uzak yemek borusunda boyunca kas üzerinde 5 mm boyuna kesi yapmak. </li><li> Aynı yönde (kesi 1, Şekil 1B) boyunca epitel açın kesin. (Kesi 2, 3 ve 4, Şekil 1B) keskin bir işletim makas ile mide çoğu kesmek için mide üç kesiler olun. Mide boş değilse, dikkatli gastrik içeriğini kaldırmak. </li><li> Bir 8-0 prolen sutu koyunYeniden doğru mukozal ile Noktası üzerinden (mide üzerine) (yemek borusu üzerine) ve Point &#39;(konik noktası iğne ile) muhalefet mukozal. Sırasıyla Nokta b ve b Nokta &#39;, ve Points diğer çiftleri aracılığıyla Aynı şekilde yer dikişler. Eşit olarak, bu dikişler alanı birbirinden 3-4 mm. </li><li> Kan ve mide içeriğini temizlemek için serum fizyolojik ile karın boşluğuna yıkayın. Ipek dikişlerle karın duvarı ve metal klipleri ile cilt kapatın. </li></ol> 3. Karışık Reflü Modeli (Şekil 1C) <ol><li> Takip gastroözofageal kavşak açığa 2.4 2.1 adımları tekrarlayın. </li><li> Yavaşça yemek borusu arkasındaki kan damarlarından özofagus dorsal yüzü ayırın. Özofagus ve kan damarları arasında küçük bir pamuk ucu geçirin. Not: Pamuk kısmen boyutunu azaltmak için ucundan çıkarılabilir. Bu pamuk uçlu yemek borusu yukarı kaldırarak ve kan damarlarını korur iki amaca hizmet eder. </li><li> İki 5 mm boyuna inc olungastroözofageal bileşke ve keskin işletim makas ile pilor komşu duodenum yakın ucunda isions her. Duodenum üzerinde kesi için, kan damarları önlemek ve anti-mezenterik sınırında yer. </li><li> Kesintiye 8-0 Prolen sütür ile muhalefeti mukozal doğru mukozal ile kesiler anastomoz. Genellikle sırt tarafında 3-4 sütür ve ön yüzünde 2-3 sütür koyun. </li><li> Pamuk ucunu sökün. Serum fizyolojik ile karın boşluğuna yıkayın ve karın duvarı ve cilt kapatın. </li></ol> 4. Duodenal Reflü Modeli (Şekil 1D) <ol><li> Karışık reflüyü üretmek için 3.4 3.1 adım izleyin. </li><li> Dikkatle arka tarafını ortaya çıkarmak için mide yukarı kaldırın. Karaciğerin bir lob karaciğer ve mide arkası arasında bağ dokuları tarafından yakalandı olabilir. Dikkatle bağ dokuları kesmek düzgün karaciğeri korumak ve yemek borusu sırt tarafında kan damarları açığa. </li><li> Bağlanır vekan damarlarının kesti. Bağlanır ve gastroözofageal bileşkede özofagus kesti. </li><li> Pilor de duodenum Arter. Bağlanır ve Mezenterium kesti. Bütün mide çıkarın. </li><li> Normal tuzlu su ile karın boşluğuna yıkayın. Karın duvarı ve cilt kapatın. </li></ol> 5. Post-cerrahi tedavi <ol><li> Ameliyat sonrasında, bir ısıtma yastığı ile vücut ısısını korumak. Sadece sternal yatma kavuştuktan sonra, geri diğer hayvanlar ile fareler yerleştirin. </li><li> Enfeksiyonu önlemek ve ağrıyı hafifletmek için antibiyotik ve analjezik verin. Baytril enjekte (10 mg / kg, ip, qd, 3 gün boyunca) enfeksiyonu önlemek ve Buprenorfin hidroklorür için (0.05 mg / kg, sc, teklif. Iki gündür) ağrı kesici olarak. Açlık ameliyat sonrası gerekli değildir. Sıvı veya yumuşak bir diyet verilebilir. </li><li> Fareler aşağıdaki belirtileri göstermeye günlük BAZDA genel sağlık durumunu (dramatik kilo kaybı&gt;% 15, app eksikliği değerlendirinetite, seslendirme, fareler euthanize, ağız, burun veya gözler, kambur jest, hareketsizlik, anormal tımar aktivitesi) dan boşaltın. </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.

Çeşitli cerrahi modeller kemiricilerde mide, duodenum ve karışık reflüyü taklit etmek kurulmuştur. Burada tarif edilen bu üç prosedür sonrası hayatta kalma makul fiyatlar ile uzun süreli deneyler için de uygundur. Cerrahi deneyime sahip bir araştırmacı uygulama kısa bir süre sonra teknik kavramak gerekir. Kanama cerrahi, karaciğer, mide, ve kan damarlarının yanlışlıkla zarar arasında bağ dokularının ayrılması sırasında karaciğer laserasyon önce anestezik intraperitoneal enjeksiyonu ile sonuçlanabilir. Bu mümkün olduğunca kaçınılmalıdır. Pilli mini koter uygunsa kanamayı durdurmak için kullanılabilir. Organları ve dokuları manipüle edildiğinde aşırı mümkün olduğunca kaçınılmalıdır germe. Cımbız uçları pamuk ile sarılabilir, ya da pamuk uçları cımbız yerine çubuklarını gibi kullanılabilmektedir. Fare özofagus iki kat ya da iki &amp; # sahiptir8220; tüpleri &quot;, dış kırmızımsı&quot; kas tüp &quot;ve iç beyazımsı&quot; epitel tüp &quot;. Bir kesik yemek borusu ile ilgili yapıldığında, her iki tabakanın, açık kesilmesi gerekmektedir. Dikişler eşit muhalefeti mukozal doğru mukozal ile aralıklı olmalıdır. Striktür dikiş arasındaki dar mesafe nedeniyle gelişebilir ise sızıntı, dikişler arasında geniş bir mesafe ile sonuçlanabilir. Ancak, darlık gıda geçişini engelleyebilir ve hafif sızıntı yerel yapışması ve fibrotik reaksiyonu ile sabit olabilir, oysa, postoperatif ölüme neden ve böylece genellikle hayatı tehdit değildir. Protokole içinde takip etmek birkaç kritik genel ilkeler vardır. (Laboratuarında yaklaşık 20-30 dk) mümkünse ameliyat süresi kısaltılmalıdır. Ameliyat öncesi ve sonrası açlık gereksizdir. Aslında, uzun süreli açlık (&gt; 1 gün) zararlı olabilir. Cerrahi en iyisini cerrah tarafından yapılır. Bir asistan fareler uyutmak ve Surg kaynağı yardımcı olabilirnik malzemeler. Cerrah düzgün mikrocerrahi eğitimi sürece bir mikroskop gerekli değildir. Aksi takdirde Cerrahi süresi daha uzun ve bu yüzden, gerekli sonrası hayatta kalma potansiyel olarak daha kötü olabilir daha olma eğilimindedir. Bu prosedürler, önceki yayınlarda kullanılmıştır ve tatmin edici sonuçlar yarattı. Vahşi tip ve genetik olarak tadil edilmiş fareler kullanılarak, NFKB ve Nrf2 yollar gastroözofageal reflü 11,12 boyunca yemek borusu epitelindeki bariyer fonksiyonunu regüle olduğunu göstermiştir. Ancak, biz bunun yerine, bu farelerde tam gelişmiş Barrett yemek borusu ya da adenokarsinomu üretmek mümkün değildi müsinöz hücreleri ve skuamöz hücreli karsinom 9 uyarılmış edildi dağınık. Fare yemek borusu ve insan yemek borusu arasındaki histolojik farklılıklar Barrett özofagus ve özofagus adenokarsinomu üzerine araştırma bu cerrahi modellerin doğal sınırlamalar olabilir. Biz bu SU inanıyorumrgical modeller GERD ilgili çalışmaları için de faydalıdır. Genlerin ve moleküler yolların fonksiyonları GÖRH moleküler mekanizmalarını anlamak ve mekanizma bazlı tedavi geliştirmek için açıklığa edilebilir. Barrett özofagus ve özofagus adenokarsinomu üretmek için kullanıldığında, genetik değişiklikler reflü cerrahisi ile birlikte ihtiyaç duyulacaktır.

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.

Çoğu fareler (&gt;% 95) ameliyat yaşayabilir. Perioperatif dönemde, önde gelen ölüm nedenleri anestezik dozda, kanama ve bilinmeyen nedenler sayılabilir. Ameliyattan sonra dört hafta,&gt; duodenal reflü mide reflü ya da karışık reflü ve&gt;% 80 farelerin% 90 fareler yaşayabilir. Bu dönemde, fareler başta özofagus darlığı ve yemek için yetersizlik ölmektedir. Bu fareler ağır stres (kambur duruş, hareketsizlik, kusma, batık gözler, seslendirme, vs.) belirtileri göstermeye ve ötenazi gerekir. Genellikle antibiyotik ve infüzyon ile tıbbi tedavi ölüyor bu fareler kaydedemezsiniz. Fareler genellikle ameliyat sonrası gram bir çift tarafından duodenal reflü ile özellikle vücut kilo. Yine de vücut ağırlığı iyileşir ve daha sonra artar. Kırk hafta ameliyat sonrası, duodenal reflü mide ve karışık reflü ve&gt;% 70 fare ile&gt;% 80 fareler yaşayabilir. DurBu dönemi ing, bazı fareler yetersiz beslenme ya da bilinmeyen nedenlerle ölebilir. Biz ameliyattan sonra 80 hafta süreyle fareler tutmuş. Genel olarak, hala oldukça aktif ve 9 sağlıklıdır. Ameliyattan sonra dört haftada, özofagus epiteli mimarisi reflü daha az ya da zarar görür. Mikroskop altında, epitel kalın olur ve papilla uzatır. Epitel hücrelerde ve iltihaplı hücrelerde (örneğin, nötrofil, eozinofil, mast hücreleri ve) infiltrasyonunun hiperproliferasyon (Şekil 2) özofajit belirgin işaretler. Özofagus epiteli (örn IL1β, IL6 ve IL8) &#39;de Sitokinler, özellikle duodenum reflü 11 varlığında yükselir. Transmisyon elektron mikroskobu altında, dilate hücrelerarası boşluk epitel bariyer fonksiyonunun 12 bozukluğunu yansıtır. İşlev-bazlı, trans-epitelial elektrik direnci, özellikle duoden varlığında azalmış olan büyük ölçüdeal geri akış. <img alt="Figür 1" src="/files/ftp_upload/53012/53012fig1.jpg" /> Şekil 1: (A), farelerde (önden ve yandan görünüm) üst sindirim sistemi anatomisi; (B) Gastrik reflü (yandan görünüm); (C) Karışık reflü (önden görünüm); (D) Duodenal reflü (önden görünüm). <img alt="Şekil 2," src="/files/ftp_upload/53012/53012fig2.jpg" /> Şekil 2: Fare yemek borusu epitelindeki Histoloji geri akış (A) olmadan ya da gastrik geri akış (B), karışık geri akış (C), duodenal geri akış (D). Ölçek çubuğu 50 mikron =.

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.

Fare ile Gastroözofageal Reflü Cerrahi Modelleri

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

Nanyang Technological University

Research

JoVE Journal

Medicine

10.0K Görüntüleme.  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.

Video: 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.

Meatoplasty meatal stenoz Cerrahi Yönetim

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

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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.

EKG, Nabız, Core Vücut Sıcaklığı ve Serbest hareket eden Laboratuar Fare Etkinlik Veri Verimli Radiotelemetry Vericiler implantasyonu

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.

Non-invaziv Hipertermi Kanser Tedavisi için Altın Nanoparçacıklannm ve Biyolojik Sistemleri ile Radyofrekans Etkileşimleri değerlendirilmesi için Protokoller

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.

Farelerde Ateroskleroz Karşı miyointimal hiperplazide uyaran: İki Geçerli Modellerinin An Introduction

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).

Farelerde Miyokard İnfarktüsü Oluşturan Minimal İnvaziv Cerrahi Prosedür

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.

Derin Beyin Uyarım ile Saccadometry kullanarak normal ve patolojik beyin fonksiyonlarını incelemek için

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.

Deri Cerrahi denervasyon: Cilt Hastalıkları Fare modelleri Sinirler için Gereği Test Bir Yöntem

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.

Farelerde Orta Serebral Arter Tıkanıklığı ve Reperfüzyon Kaynaklı İnme Cerrahi Yaklaşım

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.

Tümör ilişkili damarlara kullanmanın intravital mikroskobu gelişmiş Dorsal deri kıvrım pencere Chambers transgenik floresan fareler üzerinde

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.

Vitro Lipid tabanlı nano tanecikleri tarafından canlı hücre Chemotherapeutics keşfetmek için Imaging teslim kullanma

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