현재의 연구는 미국 국립 보건 연구소 기금의 R01-HL092188, R01-DK083385 및 R01HL098294, 그리고 아나스타샤 교육 및 연구 재단에 의해 지원되었다하면 (FAER) HKE 및 DFG (독일 Forschungsgemeinschaft) 연구 원정대 (에게 학자금 GR2121/1- 1) AG로 미국 심장 협회 그랜트.

일반 발언. 모든 작업은 직립 해부 현미경 (Leica) 아래와 외과 coagulator를 사용하여 수행해야합니다. 실험 및 사기 집단의 쥐들이 결과의 comparability을 보장하기 위해 연령과 체중에 일치해야합니다. 온도, 혈압, 마취 및 유체 관리는 안정적이고 전체 실험 기간 동안 모니터링해야합니다. 1. 마취 및 수술 <ol><li> C57BL / 6 마​​우스 10~16주의 연령 범위 내에 있어야합니다 그리고 그들은 성별 - 검색된 각각의 그룹에 있어야합니다. </li><li> pentobarbital 나트륨 (70 밀리그램 / kg 체중의 IP)로 생쥐를 마취 약으로 마취를 유지합니다. 10 MG / ㎏ / H 나트륨 pentobarbital. 치사량에 도달 크게 혈압을 낮출 수 있으며 따라서 결과를 변경할 수 있습니다. </li><li> 직장 체온계 프로브와 온도 조절 온수 테이블 (RT, Effenberg, 뮌헨, 독일)의 장소 마우스 열 피드백에 부착된37시 체온을 유지하기 위해 컨트롤러 ° C. </li><li> 이동식 테이프와 함께 테이블에 부착된 위턱과 아래턱에 사지와 수술 테이블에 위로 향한 위치에 플레이스 마우스,. </li><li> 생존 실험의 경우 머리카락 클리핑, iodophors (예 : Betadine), autoclaved 악기 멸균 처리, 그리고 마스크에게 의사를위한 멸균 장갑을 사용으로 피부의 소독을 포함한 적절한 무균 기법이 필요합니다. </li></ol> 2. 포털 트라이 어드 폐색 <ol><li> 피부를 절단하기 때문에 마우스 헤어 알레르기의 위험을 방지하여 마우스 머리카락 흡입을 방지하기 위해 미네랄 오일과 함께 절개 영역을 커버. 당신이 쉽게 간을 노출될 수 있도록 최대 상부 끝까지 linea 알바의 중간선 개복술과 절개를 수행합니다. 가능하면은 출혈을 방지하기 위해 응고 전극 (Erbe, ICC50, Tuebingen, 독일)와 절개를 수행합니다. </li><li> 꼬리가 direc이라는 위치에 마우스로 가열 테이블을 놓으십시오외과 의사로 테드. 그 위치는 깡패를 집중하고 봉합을 배치하는 가장 좋은 설정을 제공합니다. </li><li> 포털 깡패를 폭로 젖은 솜 팁 면봉을 사용하여 오른쪽에 소장을 치환. </li><li> 약간 피임기구에 대한 평균과 왼쪽 엽과 소장에 대한 권리 엽을 바꾸다. 그게 당신에게 오른쪽, 왼쪽 / 중간 엽 (叶)의 분기점 위 포털 트라이 어드의 명확한 전망을 제공합니다. </li><li> 조심스럽게 포털 깡패를 검사하고 개성이 집게로 조심스럽게 내려 오른쪽 귓불을 누르고 있습니다. 그게 당신에게 왼쪽 포털 트라이 어드에서 멋진 전망을 제공합니다. </li><li> 왼쪽에서 오른쪽으로 간장 동맥, 간장 정맥, 그리고 담즙 덕트 포함하여 왼쪽 포탈 깡패 밑에 바늘 홀더, 경유, 일단 시각적으로 봉합사 (Ethicon, Norderstedt, 독일 7 / 0 나일론 봉합사)로 장소에게 바늘을 식별 마우스. </li><li> 스티치는 깡패 아래 긴밀하게 될 수 있습니다. 깊은 스티치는 아래에있는 베나 휴가 열등 다칠 수깡패. 따라서 바늘을 배치하기 전에 대정 맥을 식별하는 데 포셉으로 오른쪽 간 엽 (叶)을 누르십시오. </li><li> 집게와 함께 오른쪽에있는 바늘의 팁을 가지고 간 손상없이 간 영역의 밖으로 부드럽게 당기십시오. </li><li> 봉합 Eppendorf 튜브 (물이 담겨있는 약 3g)의 각 끝에 부착하고 두 봉합은 봉합 소지자 이상 종료 놓으십시오. 봉합사는 바로 트라이 어드에 방해가되지 않도록 distally 바늘의 봉합 홀더를 놓습니다. </li><li> 가중치를 적용하여 트라이 어드는 간을의 왼쪽과 중간 엽 (叶)에 대한 혈액 공급의 중단을 초래 즉시 occluded 될 것입니다. 성공적인 폐색 쉽게 허혈성 엽 (叶)의 창백 창백 시각적으로 검사하여 확인할 수 있습니다. 매달려 무게가 기둥에서 발표되고 간장이 reperfused 때 대조적으로, 색상의 변화가 즉시 사라집니다. 이 부분 간장 국소 빈혈 모델 bloo을 보존하여 mesenteric 혼잡을 피할 수우측 간 엽 (叶)으로 D 흐름. </li><li> 복강에 소장을 교체하십시오. 37.0에서 물이 흥건히 젖은 면봉으로 간, 대장의 따뜻한 유지 ° C. 수술을 마치고 상용 식품 포장지에 쥐 커버 체온이 안정 유지하는 이외에. </li><li> 실험 프로토콜에 따라 필요에 따라 국소 빈혈을 수행합니다. 정의된 국소 빈혈 시간 릴리스 후 평균과 왼쪽 간 엽 (叶)의 무게와 reperfusion가 즉시 시작됩니다. 이 기술은 생쥐에서 가역 국소 빈혈을 제작하실 수 있습니다. 더 이상의 국소 빈혈 시간이 수행되지 않을 경우주의 깊게 봉합사를 제거합니다. </li><li> 15 분 전에 직접 reperfusion 후 복강으로 약 400 μl 나트륨 염화물을 추가합니다. reperfusion 시간 동안 유체 손실을 보상하기 위해 100 μl의 금액에서 30 분 간격으로 액체를 대체할 계속합니다. </li><li> 생존 실험의 경우 수술 상처는 지속적인 근육 벽 봉합과 피부 WI를 사용 닫힙니다4 / 0 봉합 일. </li></ol> 3. 간 관류의 변화 다음과 같은 결과 매개 변수 간 부상 다음 간장 국소 빈혈을 결정하기 위해 권장합니다 : 간 효소 (예 : ALT, 대서양 표준시)와 간 조직학 (H &amp; E 염색법) 다음과 같은 간 국소 빈혈과 reperfusion 3,9를 결정합니다. <img alt="그림 1" src="/files/ftp_upload/2550/2550fig1.jpg" /> 그림 1 간 국소 빈혈에 대한 교수형 - 무게 시스템의 모델 :. 매달려 체중 시스템이 왼쪽과 중간 엽 3 간 국소 빈혈을 유발하기 위해 설립되었습니다. () 따라서 꼬리가있는 오른쪽 엽은 부드럽게 왼쪽 엽에서 분리되었다. 바로 기억 상실증이 분명 오른쪽 중간 좌우 엽 (叶)의 분기점 위 포털 깡패를 보려면 다음 약간 이동했습니다. 8.0 나일론 봉합사는 왼쪽 간장 깡패 아래에 배치되었다. (B) 각각의 봉합사의 끝에 작은 무게 (시 3g Eppendorf 부엉물이 가득 bes)을 첨부와 적용되었습니다. 간장 국소 빈혈이 빨간색에서 연한에 왼쪽과 중간 엽의 색깔 변화로 볼 수 있습니다. 이 실험 설계는 체중 부하를 적용하고 방출하여 정확한 폐색과 왼쪽 간장 트라이 어드의 reperfusion 수 있습니다. <img alt="그림 2" src="/files/ftp_upload/2550/2550fig2.jpg" /> 그림 2. 알라닌 (ALT)와 aspartate (대서양 표준시) 생쥐의 아미노 전이 효소에 따라 달라질 간 국소 빈혈 시간의 영향. 일부 포털 트라이 어드의 국소 빈혈은 지시 (- 60 분 국소 빈혈 0)로 유도되었다. reperfusion의 2 시간 후, ALT ()와 대서양 표준시 (B)가 측정되었다. 7 생쥐 / 그룹 - 결과가 수단 ± SE 6으로 표현됩니다. * P 0 분 국소 빈혈 (가짜 제어)에 비해 &lt;0.05.

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+HIF-2+regulates+erythropoietic+responses+to+hypoxia+in+renal+anemia.">Hepatic HIF-2 regulates erythropoietic responses to hypoxia in renal anemia.</a> Blood. , (2010).</li></ol>

관심의 어떠한 충돌 선언 없습니다.

현재 연구는 왼쪽 간장 포털 트라이 어드의 폐색에 대한 교수형 - 무게 시스템을 사용하여 murine 모델에서 국소 빈혈을 수행하는 방법을 설명합니다. 그것 배우는 쉬운 방법이지만 다음과 같은 함정은 고려되어야한다. 처음에 아주 흔한 실수는 포털 깡패와 대정 맥을 포함하고 깊은 스티치입니다. 호흡과 심장 박동이 증가 저항 및 생쥐의 초기 사망의 결과로 심장에 혈액을 반환의 감소로 인해 15 분 내에 진정됩니다. 대정 맥의를 포함을 방지하는 가장 좋은 방법은 집게로 바로 엽 (叶)을 누른 스티치 전에 대정 맥을 집중하는 것입니다. 또 다른 중요한 문제는 가중치도 오른쪽 엽 (叶)으로 혈액의 흐름을 중지 왼쪽 및 오른쪽 포털 시범 서비스에 너무 많은 스트레스를 초래할 수있는 무거운에없는 것입니다. 따라서 Eppendorf 튜브에있는 물의 양은 마우스 무게와 s에 조정되어야합니다uture 보유자는 스티치 위치에서 distally 배치해야합니다. 반대편에서 가중치가 낮은 혈압에있는 경우는 완전히 중지되지 않습니다. 두 실수는 쉽게 각각 중간과 왼쪽 엽 (叶)의 오른쪽 귓불이나 부족한 색상 변화의 창백한에 추가 색상의 변화가 감지된다. 따라서 가중치를 적용 따라 간장 색 변화는 10 분마다 제어되어야합니다. 또 다른 일반적인 수술 합병증은 바늘로 간 부상이다. 따라서 오른쪽 엽 (叶)은 바늘 끝이 쉽게 볼 수와 바늘 홀더로 찍은 수 있도록 개성이 집게로 잡고해야합니다. 처음에 이러한 함정을 고려하는 경우에는 간장 국소 빈혈에 대한 보내야-체중 시스템은 포털 트라이 어드의 클램핑과 관련된 간 손상의 변화를 최소화하여 국소 빈혈에 의한 높은 재현성 부상을 제공합니다. 교수형 - 무게 시스템을 사용하여 포털 깡패는 단지 전체에 걸쳐 한번에 조작할 수 있습니다수술은 간장 엽 (叶)에 훨씬 적은 피해를 입었습니다. 또한, 아무런 간장이나 장내 혼잡이 기술로 발생하지 않습니다. 더 많은 조직 외상, 특히 허혈성 preconditioning 동안 같은 여러 국소 빈혈 사이클 동안 클램프의 reapplication으로, 선박 클램프를 제거하고 교체에 의해 발생합니다. 클램프의 reapplication에 의한 간 엽 (叶)의 조작에 의한 조직 손상을 방지하면서, 우리의 관찰에 의하면, 간 조직에서 원격 위치에 가중치를 어슬렁의 사용은 안정적인 폐색의 장점을 제공합니다. 함께 찍은, 현재의 연구는 클램핑과 관련된 다양성과 한계를 최소화, 국소 빈혈 동안 포털 깡패 폐색에 대한 보내야-체중 시스템의 가능성을 제공합니다. 따라서이 기법은 간장 국소 빈혈을 공부 고려해 다른 수사에 대한 관심 있을겁니다.

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> Pentobarbital 나트륨 (치명​​ 플러스) </td><td> Vortech 제약 저게 공사 </td><td> VPL 9372 </td><td> 식염수 4 밀리그램 / ML </td></tr><tr><td> 봉합사, Prolene 8.0 </td><td> Ethicon, 미국 </td><td> M8739 </td><td> 재사용 </td></tr><tr><td> 현미경 해부 </td><td> Leica </td><td> N / A </td><td> 관대한 작동 거리를 고려 </td></tr><tr><td> 가열 표 </td><td> RT, Effenberger, 독일 </td><td> N / A </td><td> 전용 및 단일 공급 업체 </td></tr><tr><td> 외과 용구 </td><td> WPI, 뒤몽 </td><td></td><td> 포셉, 니들 홀더 </td></tr><tr><td> Coagulator </td><td> Erbe, ICC50, Tuebingen, 독일 </td><td> N / A </td><td> coagulators의 다른 업체 </td></tr></tbody></table> 표 1. 특정 시약 및 장비.

use of a hanging-weight system for liver ischemia in mice

국소 빈혈로 인한 급성 간 손상은 여러 임상 절차 예 : 간 이식, 간장 종양의 절제술 또는 외상 수리하는 동안 발생할 수 있으며, 높은 사망률 1-2가 간경화가 발생할 수 있습니다. 간장 국소 빈혈에 따라서 murine 연구 pharmacological 및 유전자 연구에게 3-9를 활용할 수있는 기회를 제공함으로써 연구의 중요한 분야가되었습니다. 특히, 유전자의 조직이 구체적인 삭제와 조건부 생쥐 (cre, flox 시스템)은 특정 조직 10-13에서 단백질의 역할에 통찰력을 제공합니다. 때문에 수동으로 생쥐에서 포털 깡패를 클램핑과 관련된 기술적인 어려움으로 인해 우리는 이전에 3을 설명했습니다 포털 트라이 어드의 폐색에 대한 교수형 - 무게 시스템을 사용하여 체계적인 평가를 수행했습니다. 교수형 - 체중 시스템을 사용함으로써 우리는 또한 이후 간장 엽 (叶) 손상을 유발하지 않고도 포털 트라이 어드의 왼쪽 지점 주위 봉합 배치가능한 최고의 클램프로 인해 혈관에 간 조직의 가까운 위치에 간장 조직의 손상을 일으킬 수 있습니다. 더욱이, 간성 트라이 어드의 오른쪽 지점은 여전히​​ 따라서 perfused 더 창자 정체 보존되며 오른쪽 간장 엽 (叶)으로 혈액의 흐름으로이 기법으로 발생하지 않습니다. 또한, 포털 깡패는 전체 수술에 걸쳐 한번에 조작할 수 있습니다. 따라서 절차를 쉽게 수행할 수 있으며, 국​​소 빈혈과 reperfusion의 짧은 시간으로 미리 에어컨을 좋아해요. 알라닌 (ALT)와 aspartate (대서양 표준시) 아미노 전이 효소 혈청 수준 3,9으로 측정으로 다양한 국소 빈혈과 reperfusion 시간을 수행하여이 모델의 체계적인 평가는 간 손상과 간장 국소 빈혈 시간 가까운 상관 관계를 밝혔다. 간장 국소 빈혈과 간헐 reperfusion 위해 보내야-체중 시스템을 사용할 때 함께 찍은,이 연구는 높은 재현성 간 부상을 확인합니다. 따라서이 기법은 다른 수사관에 관심을 가져 주셔서 유용할 수 있습니다생쥐의 간 국소 빈혈 연구의 에드. 따라서 비디오 클립이 기술에 대한 자세한 단계별 설명을 제공합니다.

Watch this Scientific Journal Video about Use of a Hanging-weight System for Liver Ischemia in Mice at JoVE.com

Use of a Hanging-weight System for Liver Ischemia in Mice

우리는 포털 깡패 폐색에 대한 교수형 체중 시스템의 소설 murine 모델을 설립했습니다. 이 기술은 murine 간장 모델에서 국소 빈혈의 향후 수사 유용할 수 있습니다.

마우스의 간 국소 빈혈에 대한 교수형 - 체중 시스템의 사용

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and ...

use-of-a-hanging-weight-system-for-liver-ischemia-in-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

15.9K 조회수.  University of Colorado, Denver. 우리는 포털 깡패 폐색에 대한 교수형 체중 시스템의 소설 murine 모델을 설립했습니다. 이 기술은 murine 간장 모델에서 국소 빈혈의 향후 수사 유용할 수 있습니다.

비디오: Use of a Hanging-weight System for Liver Ischemia in Mice

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. Cardioprotection by ischemic preconditioning (IP) remains an area of intense investigation. To further elucidate its molecular basis, the use of knockout mouse studies is particularly important 7, 14, 30, 39. Despite the fact that previous studies have already successfully performed cardiac ischemia and reperfusion in mice, this model is technically very challenging. Particularly, visual identification of the coronary artery, placement of the suture around the vessel and coronary occlusion by tying off the vessel with a supported knot is technically difficult. In addition, re-opening the knot for intermittent reperfusion of the coronary artery during IP without causing surgical trauma adds additional challenge. Moreover, if the knot is not tied down strong enough, inadvertent reperfusion due to imperfect occlusion of the coronary may affect the results. In fact, this can easily occur due to the movement of the beating heart.
Based on potential problems associated with using a knotted coronary occlusion system, we adopted a previously published model of chronic cardiomyopathy based on a hanging weight system for intermittent coronary artery occlusion during IP 39. In fact, coronary artery occlusion can thus be achieved without having to occlude the coronary by a knot. Moreover, reperfusion of the vessel can be easily achieved by supporting the hanging weights which are in a remote localization from cardiac tissues.
We tested this system systematically, including variation of ischemia and reperfusion times, preconditioning regiments, body temperature and genetic backgrounds39. In addition to infarct staining, we tested cardiac troponin I (cTnI)
as a marker of myocardial infarction in this model. In fact, plasma levels of cTnI correlated with infarct sizes (R2=0.8). Finally, we could show in several studies that this technique yields highly reproducible infarct sizes during murine IP and myocardial infarction6, 8, 30, 40, 41. Therefore, this technique may be helpful for researchers who pursue molecular mechanisms involved in cardioprotection by IP using a genetic approach in mice with targeted gene deletion. Further studies on cardiac IP using transgenic mice may consider this technique.

A murine model for myocardial ischemia and ischemic preconditioning is an important tool study cardioprotective mechanisms in vivo. Here, we report an easy applicable in situ model for cardiac IP using a hanging-weight system for coronary artery occlusion.

마우스의 관상 동맥 폐색에 대한 한깅 무게 시스템의 사용

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. ...

연구

의학

Use of a Hanging-weight System for Isolated Renal Artery Occlusion

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients revealed that only a mild increase in serum creatinine levels (0.3 to 0.4 mg/dl) is associated with a 70% greater risk of death than in persons without any increase 1. Along these lines, surgical procedures requiring cross-clamping of the aorta and renal vessels are associated with a renal failure rates of up to 30% 4. Similarly, AKI after cardiac surgery occurs in over 10% of patients under normal circumstances and is associated with dramatic increases in mortality. AKI are also common complications after liver transplantation. At least 8-17% of patients end up requiring renal replacement therapy 5. Moreover, delayed graft function due to tubule cell injury during kidney transplantation is frequently related to ischemia-associated AKI 6. Moreover, AKI occurs in approximately 20% of patients suffering from sepsis 6.The occurrence of AKI is associated with dramatic increases of morbidity and mortality 1. Therapeutic approaches are very limited and the majority of interventional trials in AKI have failed in humans. Therefore, additional therapeutic modalities to prevent renal injury from ischemia are urgently needed 3, 7-9. 
To elucidate mechanisms of renal injury due to ischemia and possible therapeutic strategies murine models are intensively required 7-13. Mouse models provide the possibility of utilizing different genetic models including gene-targeted mice and tissue specific gene-targeted mice (cre-flox system). However, murine renal ischemia is technically challenging and experimental details significantly influence results. We performed a systematic evaluation of a novel model for isolated renal artery occlusion in mice, which specifically avoids the use of clamping or suturing the renal pedicle 14. This model requires a nephrectomy of the right kidney since ischemia can be only performed in one kidney due to the experimental setting. In fact, by using a hanging-weight system, the renal artery is only instrumented once throughout the surgical procedure. In addition, no venous or urethral obstruction occurs with this technique. We could demonstrate time-dose-dependent and highly reproducible renal injury with ischemia by measuring serum creatinine. Moreover, when comparing this new model with conventional clamping of the whole pedicle, renal protection by ischemic preconditioning is more profound and more reliable. Therefore his new technique might be useful for other researchers who are working in the field of acute kidney injury.

A precise murine model for acute kidney injury (AKI) due to ischemia is an important tool to investigate acute kidney injury and possibly find therapeutic tools to treat renal injury. The hanging weight system offers a tool for immediate and reliable renal artery occlusion and reperfusion without causing renal congestion.

절연 신장 동맥 폐색에 대한 한깅 - 무게 시스템의 사용

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

소설 허벌 요법을 평가하는 패혈증의 동물 모델의 사용

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Intraoperative Detection of Subtle Endometriosis: A Novel Paradigm for Detection and Treatment of Pelvic Pain Associated with the Loss of Peritoneal Integrity

Endometriosis is a common disease affecting 40 to 70% of reproductive-aged women with chronic pelvic pain (CPP) and/or infertility. The purpose of this study was to demonstrate the use of a blue dye (methylene blue) to stain peritoneal surfaces during laparoscopy (L/S) to detect the loss of peritoneal integrity in patients with pelvic pain and suspected endometriosis. Forty women with CPP and 5 women without pain were evaluated in this pilot study. During L/S, concentrated dye was sprayed onto peritoneal surfaces, then aspirated and rinsed with Lactated Ringers solution. Areas of localized dye uptake were evaluated for the presence of visible endometriotic lesions. Areas of intense peritoneal staining were resected and some fixed in 2.5% buffered gluteraldehyde and examined by scanning (SEM) electron microscopy. Blue dye uptake was more common in women with endometriosis and chronic pelvic pain than controls (85% vs. 40%). Resection of the blue stained areas revealed endometriosis by SEM and loss of peritoneal cell-cell contact compared to normal, non-staining peritoneum. Affected peritoneum was associated with visible endometriotic implants in most but not all patients. Subjective pain relief was reported in 80% of subjects. Based on scanning electron microscopy, we conclude that endometrial cells extend well beyond visible implants of endometriosis and appear to disrupt the underlying mesothelium. Subtle lesions of endometriosis could therefore cause pelvic pain by disruption of peritoneal integrity, allowing menstrual or ovulatory blood and associated pain factors access to underlying sensory nerves. Complete resection of affected peritoneum may provide a better long-term treatment for endometriosis and CPP. This simple technique appears to improve detection of subtle or near invisible endometriosis in women with CPP and minimal visual findings at L/S and may serve to elevate diagnostic accuracy for endometriosis at laparoscopy.

Loss of peritoneal integrity provides a new paradigm to understand and treat chronic pelvic pain in women with mild forms of endometriosis and can be easily detected using intraoperative instillation of dye at the time of laparoscopy.

미묘한 Endometriosis의 Intraoperative 감지 : 복막 무결성의 손실과 관련된 골반 통증의 감지 및 치료를위한 소설 패러다임

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

쥐의 척수 손상의 모델로 초점 국소 빈혈을 Photothrombosis는 유도

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

마우스 및 쥐에 대한 낮은 흐름 디지털 마취 시스템의 사용

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality worldwide. In humans, a pathologic occlusion forms in a coronary artery and reperfusion of that occluded artery is considered essential to maintain viability of the myocardium at risk. Although essential for myocardial recovery, reperfusion of the ischemic myocardium creates its own tissue injury. The physiologic response and healing of an ischemia/reperfusion injury is different from a chronic occlusion injury. Myocardial ischemia/reperfusion injury is gaining recognition as a clinically relevant model for myocardial infarction studies. For this reason, parallel animal models of ischemia/reperfusion are vital in advancing the knowledge base regarding myocardial injury. Typically, ischemia of the mouse heart after left anterior descending (LAD) coronary artery occlusion is confirmed by visible pallor of the myocardium below the occlusion (ligature). However, this offers only a subjective way of confirming correct or consistent ligature placement, as there are multiple major arteries that could cause pallor in different myocardial regions. A method of recording electrocardiographic changes to assess correct ligature placement and resultant ischemia as well as reperfusion, to supplement observed myocardial pallor, would help yield consistent infarct sizes in mouse models. In turn, this would help decrease the number of mice used. Additionally, electrocardiographic changes can continue to be recorded non-invasively in a time-dependent fashion after the surgery. This article will demonstrate a method of electrocardiographically confirming myocardial ischemia and reperfusion in real time.

During murine myocardial ischemia/reperfusion surgery, correct placement of the occluding ligature is typically confirmed by visible observation of myocardial pallor. Herein, a method of electrocardiographically confirming ischemia and reperfusion, to supplement observed myocardial pallor, is demonstrated in male C57Bl/6 mice.

표면 패드 심전도를 사용하여 마우스의 심근 허혈 및 재관류 손상의 확인

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality ...

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness 1,3,4,5. Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn&#39;s disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNF&#916;RE6. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.

Murine studies in models of colonic inflammation have demonstrated that a small percentage (1 - 5%) of mesenchymal stem cells (MSC) injected intravenously or intraperitoneally home to the inflamed colon1,2. This study shows that ultrasound-guided intracardiac injections of MSCs result in increased localization to the intestine.

초음파 유도 Intracardiac 주입 실험 선 동적인 장 질병의 Murine 모델에 사용 하기 위해 내장을 추적 증가 인간 중간 엽 줄기 세포의

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

뮤린 정밀 절단 간 슬라이스 간 생물학의 Ex Vivo 모델로

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...