저자는이 실험 모델의 개발을 지원하기 위해 각각 운영 교부금 및 설립 기금을위한 의학 연구에 대한 건강 연구 (MOP53116)와 앨버타 헤리티지 재단의 캐나다 연구소에 감사드립니다.

1. 마취 <ol><li> 2L/min의 마취 기계의 흐름 속도를 설정합니다. 진공 흡입에 배기를 연결합니다. </li><li> 5 % 마취 가스 (Isoflurane) (~ 3 분)로 요금 얼굴 마스크. </li><li> 신생아 piglets 100 % 산소 (~ 3 분)에 흡입 Isoflurane 5퍼센트으로 유도 될 것입니다. </li><li> Isoflurane로부터 2-3 %로 마취를 유지합니다. 적절한 0.5 % Isoflurane의 미세한 조절은 그러나 0.5 5 %에​​서 piglets의 상태에 따라 다양 할 수 있습니다. </li><li> 혈관 액세스가 설립되면 inhalational 마취는 펜타닐 (5-50 mcg / kg / H)와 midazolam (200-500 mcg / kg / H) 혼합제를 사용하여 정맥 주사 마취로 전환 할 수 있습니다. Pancuronium (50-100 mcg / kg / H)는 마취 약품의 조정에 보존되어 동물의 상태를 관찰 할 수있는 능력을하면서, 수술 중에 과도한 근육의 움직임을 제어해야 할 수 있습니다. </li><li> 돼지는 (percutaneous 산소의 펄스 oximetry에 의해 모니터링된다95-100%에서 aturation)와 ECG (130-170 비트 / 분의 심장 박동). </li><li> 돼지의 직장 온도는 38-40에서 관리하고 있습니다 난방 담요 찬란한 따뜻한 갖춘 ° C. </li><li> 돼지의 마취 상태를 정기적으로 행동 (동요), 유산소 운동 (빈맥과 고혈압)과 호흡 (tachypnoea) 매개 변수가 적절한, 신경 (학생 크기, 찢어 몸의 움직임)를 사용하여 실험 기간 동안 평가되고있다. 최소한의 마비가 제공됩니다. 마비로와없는 piglets의 마취의 이전 경험을 평가하는 데 유용 할 것입니다. </li><li> 프로토콜은 pentobarbital (100 밀리그램 / kg) intravenously의 과다 복용과 실험의 끝 부분에있는 동물의 euthanization과 비 생존 절차입니다. </li></ol> 2. 사타구니의 혈관 카테터의 수술 배치 (그림 1) <ol><li> 오른쪽 사타구니 긴 2~3센티미터 절개를합니다. </li><li> 오른쪽 넙 다리 정맥과 1cm 맞의 1cm를 해부t 대퇴 동맥. 각 선박 통해 두 3-0 문자열을 놔. </li><li> 오른쪽 넙 다리 정맥 catheterization : 정맥의 말초을 Ligate. 15cm에 아가일 카테터 (3.5 ~ 5, 프랑스어, 더블 루멘) (Covidien, 맨스필드, MA)를 삽입하고이 오른쪽 심방에 게재합니다. 카테터를 확보하려면 두 문자열을 묶어. 카테 테르는 유지 보수 유체 및 약물 주입 (보조 포트) 및 정맥 / 오른쪽 심방 압력 측정 (기본 포트) 중앙 사용할 수 있습니다. </li><li> 오른쪽 넙 다리 동맥 catheterization : 동맥의 말초을 Ligate. 혈액의 흐름을 중지 근위 문자열을 올려요. 5cm에 아가일 카테터를 (3.5 ~ 5, 프랑스어, 단일 루멘)를 넣습니다. 이 연속 평균 동맥 압력 측정 및 혈액 샘플링에 대한 인프라 신장 대동맥에 동맥 카테터를 삽입합니다. 카테터를 확보하려면 두 문자열을 묶어. </li><li> 피부를 닫습니다. </li></ol> 3. 기계 환기 (그림 2) 수립 <ol><li> 긴 2~3cm 수평을목에 절개. </li><li> 해부 및 기관의 1cm 쉽게받을 수 있습니다. 기관 통해 두 1-0 문자열을 놔. </li><li> 기관에 1cm에서 endotracheal 튜브를 (3.0 또는 3.5)를 넣습니다. 환풍기에 연결, 기계적 환기를 시작. endotracheal 튜브를 고정합니다. </li><li> 해부 및 온목 동맥을 쉽게받을 수 있습니다. 지속적으로 혈액의 흐름을 측정하는 교통 시간 초음파 흐름 프로브 (2SB 또는 2RB, 음속에 가까운 시스템 주식회사 Ithica, NY)로 배를 에워 싸다. </li></ol> 4. 장간막의 흐름 프로브의 위치 (그림 3)과 왼쪽 신장 (그림 4) 동맥 <ol><li> 펜타닐 (5-10 mcg / kg)와 acepromazine이 (0.01-0.02 밀리그램 / kg) 이전에 피부 절개를 필요의 추가 복용. </li><li> 긴 subcostal - 측면 절개를 만들어 조심스럽게 근육 레이어를 해부. </li><li> 복부 대동맥을 쉽게받을 수 있습니다. </li><li> 혈관 취급 (vasospasm)과 림프 부상을 최소화합니다. </li><li> 0.5-1cm 장간막 동맥과를 해부주위 천음속 흐름 프로브 (3SB)를 넣어. </li><li> 0.5-1cm 왼쪽 신장 동맥을 해부하고 주위에 음속에 가까운 흐름 프로브 (2SB)를 넣어. </li><li> 피부를 닫고 흐름 프로브를 보호합니다. </li></ol> 5. 폐 동맥 카테터의 위치 (그림 5)와 흐름 프로브 (그림 6) <ol><li> 펜타닐 (5-10 mcg / kg)와 acepromazine이 (0.01-0.02 밀리그램 / kg) 이전에 피부 절개를 필요의 추가 복용. </li><li> 오른쪽 측면 위치에서 동물을 누워. </li><li> 왼쪽 네번째 갈비 공간에서 Thoracotomy. </li><li> 필요한 경우 내유동 맥과 정맥은 조심, ligate. </li><li> 왼쪽 폐를 눌러 필요에 따라 산소를 증가 치과 면봉을 사용합니다. </li><li> 심낭을 엽니 다. </li><li> 동맥관 폐동맥에서 대동맥으로 실행되는을 식별합니다. </li><li> 동맥관은 클립을 배치하거나 그 기원에 두꺼운 &quot;3-O 실크&quot;타이의 출혈도 잡았 될 수 있습니다. </li><li> 무료의 주요 폐 동맥과 패스 AV두꺼운 &quot;0&quot;넥타이를 사용 ascular 슬링. </li><li> 폐 동맥 카테터의 위치에 대한 기본에서 지갑 문자열 (5-0 prolene) 봉합사를 수행합니다. </li><li> 1cm의 최대 지갑 문자열을 통해 20G Angiocath을 (카테터의 끝에서 1cm 이하 3 쪽 구멍이있는)를 넣습니다. </li><li> 정맥 혈액의 흐름을 무료로 있는지 확인합니다. </li><li> 압력 변환기에 연결 하 동맥과 폐동맥 압력 및 파형을 확인합니다. </li><li> 지갑 문자열을 조여와 폐 카테터를 확보. </li><li> 주요 폐 동맥 주위에 음속에 가까운 흐름 프로브 (6SB)를 배치합니다. </li><li> 흐름 프로브 및 최적의 신호 전달을 허용하는 동맥 사이에 초음파 젤을 삽입합니다. </li><li> 습한 생리 거즈로 상처를 다룹니다. </li></ol> 6. Hypoxia 및 reoxygenation 프로토콜 <ol><li> hypoxemia를 유도하기 위해 흡입 질소 가스의 농도를 증가하여 10 %에서 영감을 산소 농도를 감소. </li><li> 영감 산소를 조정10 % 15% 사이의 농도 2 시간에 30-40%의 20-40 mmHg 또는 SAO 2 파오 2 얻을 수 있습니다. </li><li> 파코 2 평가하고 그에 따라 환풍기 속도를 조정하는 동맥혈 분석을 수행합니다. </li><li> hypoxemia의 유도로, 첫 번째 시간은 지속적으로 tachycardic (및 심장 출력) 응답을 유도하기 위해 최선을 다하고 있습니다. </li><li> 일반적인 경동맥, 장간막과 왼쪽 신장 동맥에서 혈액의 흐름의 변화를 모니터링하고 있습니다. </li><li> hypoxia의 두 번째 시간에, hypoxic 스트레스가 기준선의 30~40%에 지속적으로 낮은 심장 출력 증가, 30-35 mmHg 및 동맥 산도 6.95-7.05에 동맥 압력을 의미합니다. </li><li> Hypoxic 스트레스가 중간에 중단하거나 적절한 15 분으로 연장 할 수 있습니다. </li><li> 순수 산소를 계속하는 동안 갑자기 100 %가 질소 가스를 중단하여 갑자기 영감을 산소 농도를 높입니다. </li><li> 동맥 압력 및 기타 hemodynamic 패러을 의미, 심장 출력을 모니터빠른 복구를 위해 거리에 있습니다. </li><li> 100 % 산소로 인공 호흡은 0.5h 동안 계속 될 수 있습니다. 이 기간 후, 21 %로 빠르게 영감을 산소 농도를 줄일 수 있습니다. </li><li> 실험의 남은 기간 동안 21 % 산소와 reoxygenation을 계속합니다. 필요한 경우 영향을받은 산소 농도는 25 % titrated 할 수 있습니다. </li><li> 10 ML / kg 벨소리의 락트산 솔루션의 유체 boluses은 실험 기간 동안 적절한 필요할 수 있습니다. 의 사용은 protocolized해야합니다. </li></ol> 7. 대표 결과 : hypoxia의 첫 번째 시간을 통해 신생아 돼지의 hypoxemia의 유도 120 % -130 기준의 % (그림 7A)과 심장 박동 (그림 7B)로 심장 출력 (폐 동맥 흐름)을 증가해야합니다. 일반적으로, 심장 출력은 첫 번째 0.5h 및 hypoxia의 1 시간 사이에 최고 보상에 도달해야합니다. 또한, 혈액 흐름의 결과로 중앙있게됩니다 저를 감소senteric 및 신장 관류하지만 보존 또는 증가 온목 동맥 흐름 (그림 8). hypoxia의 두 번째 시간 동안 발생이나 부정맥이없는 심장 박동의 저하 심장 출력의 꾸준한 감소, 저혈압의 개발 (그림 9A)이 있습니다. Hypoxia 가끔 심장 출력 감소로 hypoxia의 마지막 30 분에 낮출 수 증가 폐동맥 압력 (그림 9B)로 폐 고혈압을 유발한다. 인공 호흡하면 모든 hemodynamic 매개 변수는 즉시 점차 reoxygenation의 첫 번째 시간으로 복구 신장 혈액의 흐름을 제외하고 normoxic 기준으로 회복됩니다. 그러나, 특히 심장 출력 및 동맥 압력을 의미 hemodynamic 매개 변수가 점차 각각 normoxic 기준과 35-​​45 mmHg의 70-75 % 정도에 reoxygenation의 첫 2 시간이 넘는 저하됩니다. 이 심장 혈관 dysfuncti에는 적어도 같은 vasoactive과 inotropic 에이전트와 같은 심근 아름다운 및 보증 심장 혈관 지원 치료에 대한 부분입니다. <img alt="그림 1" src="/files/ftp_upload/3166/3166fig1.jpg" /> 그림 1 : 넙 다리 동맥과 정맥 카테터의 위치와 사타구니 절개 <img alt="그림 2" src="/files/ftp_upload/3166/3166fig2.jpg" /> 그림 2 : 온목 동맥 주위에 endotracheal 튜브의 위치와 흐름 프로브와 목을 절개 <img alt="그림 3" src="/files/ftp_upload/3166/3166fig3.jpg" /> 그림 3 : 장간막 동맥의 절연 측면 절개 <img alt="그림 4" src="/files/ftp_upload/3166/3166fig4.jpg" /> 그림 4 : 왼쪽 신장 동맥의 절연 측면 절개 <img alt="그림 5" sRC는 = &quot;/ files/ftp_upload/3166/3166fig5.jpg&quot;/&gt; 그림 5 : 폐동맥 카테터의 위치와 Thoracotomy <img alt="그림 6" src="/files/ftp_upload/3166/3166fig6.jpg" /> 그림 6 : 주 폐동맥 주위에 음속에 가까운 흐름 프로브의 위치와 Thoracotomy <img alt="그림 7" src="/files/ftp_upload/3166/3166fig7.jpg" /> 그림 7 : (A) 심장 출력 (폐 동맥 흐름)와 (B) 심장 박동 hypoxia와 reoxygenation 동안의 시간적 변화 <img alt="그림 8" src="/files/ftp_upload/3166/3166fig8.jpg" /> 그림 8 : (A) 일반적으로의 경동맥에서 혈액의 흐름에 시간적 변화, (B) 장간막 및 hypoxia와 reoxygenation 중 (C) 왼쪽 신장 동맥 <img alt="그림 9" src="/files/ftp_upload/3166/3166fig9.jpg" /> 그림 9 : 측두엽의 변화는 (A) 동맥 압력을 의미하고 hypoxia와 reoxygenation 중 (B) 폐동맥 압력

<ol>
	<li>Borke, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+myocardial+matrix+metalloproteinases+in+hypoxic+newborn+pigs+during+resuscitation:+effects+of+oxygen+and+carbon+dioxide.">Increased myocardial matrix metalloproteinases in hypoxic newborn pigs during resuscitation: effects of oxygen and carbon dioxide.</a> Eur. J. Clin. Invest. 34, 459-466 (2004).</li><li>Munkeby, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resuscitation+of+hypoxic+piglets+with+100%+O2+increases+pulmonary+metalloproteinases+and+IL-8.">Resuscitation of hypoxic piglets with 100% O2 increases pulmonary metalloproteinases and IL-8.</a> Pediatr. Res. 58, 542-548 (2005).</li><li>Haaland, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Posthypoxic+hypothermia+in+newborn+piglets.">Posthypoxic hypothermia in newborn piglets.</a> Pediatr. Res. 41, 505-512 (1997).</li><li>Haase, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resuscitation+with+100%+oxygen+causes+intestinal+glutathione+oxidation+and+reoxygenation+injury+in+asphyxiated+newborn+piglets.">Resuscitation with 100% oxygen causes intestinal glutathione oxidation and reoxygenation injury in asphyxiated newborn piglets.</a> Ann. Surg. 240, 364-373 (2004).</li><li>Stevens, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resuscitation+with+21%+or+100%+oxygen+is+equally+effective+in+restoring+perfusion+and+oxygen+metabolism+in+hypoxic+newborn+piglet+liver.">Resuscitation with 21% or 100% oxygen is equally effective in restoring perfusion and oxygen metabolism in hypoxic newborn piglet liver.</a> Shock. 27, 657-662 (2007).</li><li>Johnson, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=N-acetylcysteine+improves+the+hemodynamics+and+oxidative+stress+in+hypoxic+newborn+pigs+reoxygenated+with+100%+oxygen.">N-acetylcysteine improves the hemodynamics and oxidative stress in hypoxic newborn pigs reoxygenated with 100% oxygen.</a> Shock. 28, 484-490 (2007).</li><li>Chapados, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+cortisol+response+to+ACTH+challenge+in+hypoxic+newborn+piglets+resuscitated+with+21%+and+100%+oxygen.">Plasma cortisol response to ACTH challenge in hypoxic newborn piglets resuscitated with 21% and 100% oxygen.</a> Shock. 33, 519-525 (2010).</li><li>Cheung, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet+dysfunction+in+asphyxiated+newborn+piglets+resuscitated+with+21%+and+100%+oxygen.">Platelet dysfunction in asphyxiated newborn piglets resuscitated with 21% and 100% oxygen.</a> Pediatr. Res. 59, 636-640 (2006).</li><li>Martin-Ancel, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+organ+involvement+in+perinatal+asphyxia.">Multiple organ involvement in perinatal asphyxia.</a> J. Pediatr. 127, 786-793 (1995).</li><li>Swindle, M. M., Smith, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+anatomy+and+physiology+of+the+pig.">Comparative anatomy and physiology of the pig.</a> Scan. J. Lab. Anim. Sci. Suppl. 25, 11-22 (1998).</li><li>Chapados, I., Cheung, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Not+all+models+are+created+equal:+Animal+models+to+study+hypoxic-ischemic+encephalopathy+of+the+newborn.">Not all models are created equal: Animal models to study hypoxic-ischemic encephalopathy of the newborn.</a> Neonatology. 94, 300-303 (2008).</li><li>Liu, J. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+post-resuscitation+treatment+with+N-acetylcysteine+on+cardiac+recovery+in+hypoxia-injured+newborn+pigs.">Effects of post-resuscitation treatment with N-acetylcysteine on cardiac recovery in hypoxia-injured newborn pigs.</a> PLoS ONE. 5, e15322-e15322 (2010).</li><li>Cheung, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardio-renal+recovery+of+hypoxic+newborn+pigs+after+18%,+21%+and+100%+reoxygenation.">Cardio-renal recovery of hypoxic newborn pigs after 18%, 21% and 100% reoxygenation.</a> Intensive Care Med. 34, 1114-1121 (2008).</li><li>Temesvari, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+the+blood-brain+barrier+permeability+in+neonatal+cytotoxic+brain+edema:+laboratory+and+morphological+findings+obtained+on+newborn+piglets+with+experimental+pneumothorax.">Modulation of the blood-brain barrier permeability in neonatal cytotoxic brain edema: laboratory and morphological findings obtained on newborn piglets with experimental pneumothorax.</a> Biol. Neonate. 46, 198-208 (1984).</li><li>Domoki, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reventilation+with+room+air+or+100%+oxygen+after+asphyxia+differentially+affects+cerebral+neuropathology+in+newborn+pigs.">Reventilation with room air or 100% oxygen after asphyxia differentially affects cerebral neuropathology in newborn pigs.</a> Acta. Paediatr. 95, 1109-1115 (2006).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Part+15:+Neonatal+resuscitation:+2010+American+Heart+Association+guidelines+for+cardiopulmonary+resuscitation+and+emergency+cardiovascular+care.">Part 15: Neonatal resuscitation: 2010 American Heart Association guidelines for cardiopulmonary resuscitation and emergency cardiovascular care.</a> Circulation. 122, S909-S919 (2010).</li></ol>

관심 없음 충돌이 선언 없습니다.

현재 실험 프로토콜은 hypoxia와 reoxygenation 과정에서 신생아 과목 체계 및 지역 hemodynamic 변경 사항을 검사 할 수있는 장점이 있습니다. 우리는 또한 회복하는 동안 심장 혈관 기능을 개선하는 데 사용 개입의 각 효과를 확인할 수 있습니다. 우리와 다른 심장 혈관 1, 폐 2, neurologic 3, 위장병 4, 간장 5, 신장 6, 부신 7 hematologic 8 시스템에...

a swine model of neonatal asphyxia

질식사 관련하여 매년 백만 이상의 neonates는 전세계 죽어. 질식 neonates은 일반적으로 저혈압, 재관류 적자, hypoxic - 허혈성 뇌 병변, 폐 고혈압, vasculopathic enterocolitis, 신장 장애 및 thrombo-embolic 합병증 등의 여러 기관이 실패합니다. 동물 모델은 우리가 patho - 생리학 및 신생아 질식의 약리학을 이해하는 데 도움이하기 위해 개발하고 있습니다. 설치류 및 신생아 양에 비해, 신생아 돼지는 귀중한 모델이 될 입증되었습니다. 신생아 새끼 돼지는 36-38주의와 유사한 개발 등 여러 가지 장점을 가지고 동물의 계측 및 모니터링을 할 수 있습니다 및 혼란 변수를 제어 비교 신체 시스템, 대형 바디 크기 (출생시 ~ 1.5-2 kg)와 인간의 태아 hypoxia와 hemodynamic derangements. 여기 신생아 질식을 시뮬레이션하고 우리가 syste을 검토 할 수 있도록 실험 프로토콜을 설명마이크 및 질식 및 reoxygenation 과정뿐만 아니라 개입의 각 효과 중 지역 hemodynamic 변경됩니다. 또한, 모델은 동시에 여러 장기 장애 또는 장애를 연구의 장점과 다양한 신체 시스템과 상호 작용이 있습니다. 실험 모델은 기계 환기, 혈관 (동맥 및 중앙 정맥) 액세스의 설립을 허용하도록 신생아 piglets의 수술 기기 (1-3, 하루 과거와 1.5-2.5 kg 무게 혼합 품종)를 포함하지 않는 생존 절차입니다 와 주요 폐, 온목, 장간막과 왼쪽 신장 동맥 등 다른 동맥에서 간 혈관 압력과 혈액 흐름의 지속적 모니터링을위한 카테터와 흐름 프로브 (음속에 가까운 주식회사)의 위치. hemodynamic 매개 변수 및 일반 혈액 가스의 Z &lt;10 %의 변동에 의해 정의 된 30~60분에 대한 안정화 후,이 수술 계측의 piglets를 사용하여, 우리는 실험 프로토콜을 시작심한 hypoxemia 중은 normocapnic 폐포 hypoxia를 통해 유도된다. 돼지는 30~40%의 동맥 산소 saturations를 목표로, 2 시간에 질소 가스의 흡입 농도를 증가시켜 10-15% 산소와 통풍이 있습니다. hypoxemia의 정도는 심각한 신진 대사 산증, 전신 저혈압 및 중요한 장기 hypoperfusion있는 cardiogenic 충격으로 임상 질식을 생산합니다. hypoxia는 0.5h에 대한 100 % 산소와 다음 3.5h에 대한 21 % 산소로 reoxygenation을합니다. Pharmacologic 개입 인해 과정에서 도입하고 효과 눈이 멀어 블록 무작위 방식으로 조사 할 수 있습니다.

Watch this Scientific Journal Video about A Swine Model of Neonatal Asphyxia at JoVE.com

A Swine Model of Neonatal Asphyxia

대형 동물 모델은 신생아 질식의 생리학과 약리학의 시험에서 좋은 병진 값을 가진다. 신생아 piglets를 사용하여, 우리는 조직과 지역 혈류 역학, 생화학 및 pathologic 경로와 상관 관계가있는 산소 수송을 공부의 장점을 가지고 신생아 질식을 시뮬레이션 할 수있는 실험 프로토콜을 개발합니다.

신생아 질식의 돼지 모델

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, ...

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Medicine

14.2K 조회수.  University of Alberta. 대형 동물 모델은 신생아 질식의 생리학과 약리학의 시험에서 좋은 병진 값을 가진다. 신생아 piglets를 사용하여, 우리는 조직과 지역 혈류 역학, 생화학 및 pathologic 경로와 상관 관계가있는 산소 수송을 공부의 장점을 가지고 신생아 질식을 시뮬레이션 할 수있는 실험 프로토콜을 개발합니다.

비디오: A Swine Model of Neonatal Asphyxia

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

신생아 저산소 - 허혈성 뇌증의 돼지 모델

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

연구

의학

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

신생아 마우스 척수 압축 부상 모델

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, standardized battery of neonatal mouse motor tests. These tests can validate injury or disease models, determine treatment efficacy and/or assess motor behaviors in new transgenic strains. This paper presents a series of neonatal motor tests to evaluate general motor function, including ambulation, hindlimb foot angle, surface righting, negative geotaxis, front- and hindlimb suspension, grasping reflex, four limb grip strength and cliff aversion. Mice between the ages of post-natal day 2 to 14 can be used. In addition, these tests can be used for a wide range of neurological and neuromuscular pathologies, including cerebral palsy, hypoxic-ischemic encephalopathy, traumatic brain injury, spinal cord injury, neurodegenerative diseases, and neuromuscular disorders. These tests can also be used to determine the effects of pharmacological agents, as well as other types of therapeutic interventions. In this paper, motor deficits were evaluated in a novel neonatal mouse model of cerebral palsy that combines hypoxia, ischemia and inflammation. Forty-eight hours after injury, five tests out of the nine showed significant motor deficits: ambulation, hindlimb angle, hindlimb suspension, four limb grip strength, and grasping reflex. These tests revealed weakness in the hindlimbs, as well as fine motor skills such as grasping, which are similar to the motor deficits seen in human cerebral palsy patients.

Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

뇌성 마비의 신생아 마우스 모델에서 모터 테스트의 배터리

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, ...

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and regeneration, and to define new targets for therapeutics. For decades, models of complete heart regeneration existed in amphibians and fish, but a mammalian counterpart was not available. The recent discovery of a postnatal window during which mice possess regenerative capabilities has led to the establishment of a mammalian model of cardiac regeneration. A surgical model of mammalian cardiac regeneration in the neonatal mouse is presented herein. Briefly, postnatal day 1 (P1) mice are anesthetized by isoflurane and placed on an ice pad to induce hypothermia. After the chest is opened, and the left anterior descending coronary artery (LAD) is visualized, a suture is placed around the LAD to inflict myocardial ischemia in the left ventricle. The surgical procedure takes 10-15 min. Visualizing the coronary artery is crucial for accurate suture placement and reproducibility. Myocardial infarction and cardiac dysfunction are confirmed by triphenyl-tetrazolium chloride (TTC) staining and echocardiography, respectively. Complete regeneration 21 days post myocardial infarction is verified by histology. This protocol can be used to as a tool to elucidate mechanisms of mammalian cardiac regeneration after myocardial infarction.

This protocol describes a highly reproducible model of cardiac regeneration by surgical induction of myocardial infarction in the left ventricle of postnatal day 1 mice. The method involves induction of hypothermic anesthesia and ligation of the left anterior descending coronary artery.

신생아 쥐의 심근 경색, 심장 재생의 모델

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but regardless of the approach, an in vivo model is needed to test each treatment. The pig is one of the most used large animal models for cardiovascular disease. Its heart is very similar in anatomy and function to that of a human. The ameroid placement technique creates an ischemic area of the heart, which has many useful applications in studying myocardial infarction. This model has been used for surgical research, pharmaceutical studies, imaging techniques, and cell therapies.
There are several ways of inducing an ischemic area in the heart. Each has its advantages and disadvantages, but the placement of an ameroid constrictor remains the most widely used technique. The main advantages to using the ameroid are its prevalence in existing research, its availability in various sizes to accommodate the anatomy and size of the vessel to be constricted, the surgery is a relatively simple procedure, and the post-operative monitoring is minimal, since there are no external devices to maintain. This paper provides a detailed overview of the proper technique for the placement of the ameroid constrictor.

The purpose of this protocol is to demonstrate the placement of a delayed constricting device (an ameroid constrictor) around a coronary artery in a swine model. This device creates an ischemic area of the heart that is useful for studying new diagnostic imaging techniques and new methods of treatment.

Ameroid 압축을 사용 하 여 돼지에서 만성 심장 허 혈 모델

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

돼지에서 만성 심장 마비 모델로 tachycardia 유발 심근

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Severe Burn Injury in a Swine Model for Clinical Dressing Assessment

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological pathways impair the inflammation response, resulting in delayed wound healing. Impaired wound healing frequently occurs in patients with diabetes leading to unfavorable outcomes such as amputation. Hence, dressings having beneficial effect in promoting burn wound repair are needed. However, studies on burn wound treatment are limited due to lack of proper animal models. Our previous study demonstrated wound-healing performance in rat and swine models using a minimally invasive surgical technique. This study aimed to demonstrate a swine model of severe burn injury that eliminates wound contraction and more closely approximates the human processes of re-epithelialization and new tissue formation. This protocol provides a detailed procedure for creating consistent burn wounds and examining the wound-healing performance under the treatment of an experimental dressing in a swine model. Six burn wounds were created symmetrically on the dorsum, which were covered with a clinical dressing composed of four layers: an inner contact layer of experimental materials, an inner intermediate layer of waterproof film, an outer intermediate layer of gauze, and an outer layer of adhesive plaster. Upon the completion of experiments, wound closure, wound area, and Vancouver Scar Scale score were examined. The samples of skin resected from each animal post-sacrifice were histologically prepared and stained using hematoxylin and eosin staining. Antibacterial activity of each dressing in the context of wound healing was also examined. The application of the clinical dressing to the wounds in swine model mimics the biological processes of human wound healing with respect to the processes of epithelialization, cellular proliferation, and angiogenesis. Therefore, this swine model provides an easy-to-learn, cost-effective, and robust method to assess the effect of clinical dressings in severe burn injury.

To closely mimic the mode of burn injuries requires the interplay between clinical observation and studies in animal models. In this study, a swine model of severe burn injury was established to assess an experimental dressing in physiological and pathophysiological settings.

임상 드레싱 평가 대 한 돼지 모델에서 가혹한 화상 상해

Wound healing is a dynamic repair process and is the most complex biological process in human life. In response to burn injury, alterations in biological ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

줄기 세포 치료를위한 심근 경색 심부전의 돼지 모델 설립

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...

Myocardial Infarction by Percutaneous Embolization Coil Deployment in a Swine Model

Myocardial infarction (MI) is the leading cause of mortality worldwide. Despite the use of evidence-based treatments, including coronary revascularization and cardiovascular drugs, a significant proportion of patients develop pathological left-ventricular remodeling and progressive heart failure following MI. Therefore, new therapeutic options, such as cellular and gene therapies, among others, have been developed to repair and regenerate injured myocardium. In this context, animal models of MI are crucial in exploring the safety and efficacy of these experimental therapies before clinical translation. Large animal models such as swine are preferred over smaller ones due to the high similarity of swine and human hearts in terms of coronary artery anatomy, cardiac kinetics, and the post-MI healing process. Here, we aimed to describe an MI model in pig by permanent coil deployment. Briefly, it comprises a percutaneous selective coronary artery cannulation through retrograde femoral access. Following coronary angiography, the coil is deployed at the target branch under fluoroscopic guidance. Finally, complete occlusion is confirmed by repeated coronary angiography. This approach is feasible, highly reproducible, and emulates the pathogenesis of human non-revascularized MI, avoiding the traditional open-chest surgery and the subsequent postoperative inflammation. Depending on the time of follow-up, the technique is suitable for acute, sub-acute, or chronic MI models.

Myocardial infarction (MI) animal models that emulate the natural process of the disease in humans are crucial to understanding pathophysiological mechanisms and testing the safety and efficacy of new emergent therapies. Here, we describe an MI swine model created by deploying a percutaneous embolization coil.

돼지 모델에서 경피 색전술 코일 배치에 의한 심근 경색

Myocardial infarction (MI) is the leading cause of mortality worldwide. Despite the use of evidence-based treatments, including coronary revascularization ...