この原稿の作業は、血管造影研究所パイロットプロジェクトプログラムである止血脈管生物学（P3HVB）とAASTリサーチフェローシップによるニール博士への資金提供によって支援されました。この研究は、米国国立衛生研究所の補助金1 R35 GM119526-01およびUM1HL120877-01によって支持されている。

マウスは、12時間の明暗サイクルを有する特異的な病原体のない状態で、ピッツバーグ大学（米国ペンシルバニア州ピッツバーグ）および米国国立衛生研究所（NIH; Bethesda、MD、USA）動物ケアガイドラインに従って収容され、標準の飼料と水。すべての動物実験は、ピッツバーグ大学の動物研究およびケア委員会によって定められたガイドラインに従って承認され、実施された。 1.手術野および機器の設定<ol><li> 手順の前に、すべての外科用器具、縫合糸、ガーゼ、綿棒アプリケータ、チューブ、およびチューブコネクタを滅菌する。 <ol><li>オートクレーブ内で外科器具、縫合糸、ガーゼ、およびコットンチップアプリケータを滅菌する。エチレンオキシドでチューブとコネクターを滅菌します。 </li></ol></li><li> 手術野 <ol><li>水循環式ヒートパッドをオンにして37℃に設定します。外科医を置くその上に青色の青色のパッドを、次いで外科用青色のパッドの上に無菌のドレープを付ける。 </li><li>すべての滅菌器具を滅菌ドレープ上に開きます。滅菌手袋を使用して、この工程中の滅菌を防ぐ。 </li><li> 70％エタノールでステンレス製のボールを満たし、脇に置いてください。これは動物間の器具の清掃に使用されます。 </li><li>マイクロビーズ滅菌器をオンにし、150℃に加熱します。これは動物間の器具の清掃にも使用されます。 5匹以上のマウスで手術を行う場合は、器具を新しい無菌セットに交換してください。 </li></ol></li><li> トランスデューサのセットアップ <ol><li>滅菌トランスデューサ、PE-50チューブ、2本の23Gニードル、オス - オスのルアー、3方向ストップコックを接続します。 6 </li><li>トランスデューサを製造元の指示に従って較正してゼロにします。 </li></ol></li></ol> 2.肝切開手術<ol><li> 麻酔の誘導と位置決め </stロング&gt; <ol><li>ペントバルビタールナトリウムを70mg / kgの用量で腹腔内に注射する。麻酔は5-10分の間に有効でなければならない。つま先で麻酔深度を評価する。マウスがつま先のつまみに反応する場合、追加の時間または麻酔が必要です。処置中に追加の麻酔が必要な場合は、ペントバルビタールナトリウムを補充してください。過量を避けるために、0.05 mLよりも多い量のサプリメントを与えないでください。 </li><li>マウスが完全に麻酔下になった後、マウスを手術用ボード上に仰向けに置く。マウスの4肢すべてをテープでボードに固定します。 </li><li>剃刀で腹部と両脇の下を剃る。 </li><li>ベタディンで滅菌ガーゼを浸し、手術のために腹部および両側鼠径部に適用する。生存実験のために、腹部および鼠径部にベタジンを、次いでエタノールで合計3回の準備サイクルを準備する。 </li><li>直腸温度プローブを挿入して、手順全体を通してコア温度を監視する。 Cを保つ鉱石の温度は35〜37℃である。 </li></ol></li><li> 大腿動脈および静脈カニューレ挿入 <ol><li>静脈カテーテルの設定には、PE-10チュービング、30G針、およびIVバッグからの乳酸加リンゲル液を含む三方活栓を充填する。 </li><li>動脈カテーテルの設定：PE-10チュービングと30G針にヘパリン処理生理食塩水（1,000Uヘパリンの1:10希釈液）を充填する。凝固を防ぐにはヘパリン食塩水が必要です。 </li><li>解剖顕微鏡の下にマウスを置く。 </li><li>外科用アイリスはさみを使用して鼠径部の筋肉を4〜5mm縦切開する。デュモン鉗子を使用して、内転筋に接続された脂肪組織をつかみ、横に引っ張って大腿骨バンドルのきれいな暴露を行う。脂肪組織を切開しないでください。これにより血管損傷が起こります。 </li><li>慎重に、デュモン鉗子で動脈および静脈から神経を摘出する。神経に隣接して脂肪パッドがあります。 1つのDumont forcepでこれを取得するd横方向に引っ張る。これは動脈から神経を引き離し、解剖のための平面を作り出す。他のデュモンのforcepは、神経と動脈との間の結合組織を鈍的に切開する。 <ol><li>解剖のこの部分の間に神経をつかまえないでください。 </li></ol></li><li>大動脈周囲の3つの6-0の絹の縫合糸をループし、大腿部に近位の静脈を離す。 <ol><li>最も近位に縫合糸1を置き、緩いままにする。 </li><li>最も遠位に縫合糸2を置き、すぐに結ぶ。 </li><li>縫合糸1と2の間に縫合糸3を置き、緩くしておきます。 </li></ol></li><li>血管の腹面に動脈切開を行う。血管の切断を避けるために動脈切開術を行うには、微小血管はさみの使用を推奨します。 </li><li> 3方向カテーテルを動脈に挿入します。カテーテルを所定の位置に固定するために、直ちに縫合糸1および2を締めてください。 </li><li>三方向カテーテルをトランスデューサに接続し、ベースラインの血圧データを収集します。 </li><li>反対側の鼠径部にステップ2.2.4〜2.2.6を繰り返します。動脈と同様の方法で大腿静脈にカニューレを挿入する。血管の腹面に静脈切開を行い、続いてカテーテルを挿入する。このカテーテルは、流体投与または薬物投与に利用することができる。 </li></ol></li><li> 肝臓裂傷 <ol><li> 0.5mLのPBS、3つの吸収三角、およびマウス1匹の計量ボートを含むチューブを予め計量する。 </li><li>腹側の中線開腹を切開部から開始し、尾側に広げて肝臓を完全に露出させる。 </li><li>右の腹壁に対して腹部に吸収三角形を挿入します。左側を繰り返します。 <ol><li>肝臓が裂傷された後、詰め止めの止血効果を避けるために、吸収三角形が肝臓から離れていることを確認してください。 </li></ol></li><li>慎重に肝臓の左中葉をつかみ、外科用アイリスはさみで葉の75％を裂く。ライカレートを置くdセグメントをPBSを含むチューブに入れた。 </li><li>ステープルアプリケータでステープルで腹壁を閉めます。皮膚と筋肉を一緒につかみ、ステープルを発射する。肝臓の裂傷後、可能な限り迅速に腹部外の失血を避けるようにしてください。生存実験では、腹部は2つの層で閉じられている。筋肉のための実行可能な吸収性縫合糸に続いて皮膚のための非吸収性縫合糸の走行層が続くことにより、適切な閉鎖が提供される。 </li><li> 30分より長い生存時間のマウスでは、大腿カテーテルを除去し、動脈および静脈をステップ2.2.6の縫合糸3で結ぶ。次に、前のステップで説明したように、両側の鼠径部を2つの層で閉じる。 </li><li>出血のための特定の時間（30分〜72時間）の後、ステープルを取り除く。吸収三角形を取り除き、対応する事前に計量した計量ボートに入れます。追加の吸収三角形を使用して、吸収されていない血液を吸収してください。 </li><li&gt;吸収フィルターを秤量して総失血量を算出する。 </li></ol></li><li> 術後ケア <ol><li> 30分で外科用ボード上で屠殺し、絶え間ない監視下で、完全麻酔下で犠牲になるまでマウスを放置する。マウスは、心臓穿刺と吸入イソフルランの過剰摂取の組み合わせで安楽死させる。 </li><li>より長い生存時間ポイントのマウスを、水循環式加熱パッドの上にある回収ケージに置く。回復中にマウスを絶えず監視し、胸骨臥位を維持する意識を回復するまで無人で放置しないでください。麻酔から回復した後にのみ、他のマウスと一緒にケージ空間にマウスを戻します。 </li><li>麻酔から覚醒し、犠牲の時まで12時間ごとに皮下注射により術後鎮痛を0.1mg / kgブプレネックスで投与する。 </li><li>マウスが食物と水に自由にアクセスできるようにする。術後の正常なケージ。 </li><li>生存マウスの屠殺時に、吸入イソフルランで麻酔を行う。麻酔下になったら、右心臓の心臓穿刺により血液が採取され、上記のように失血が記録され、最後に安楽死がイソフルランの過剰投与により保証される。 </li></ol></li></ol>

<ol>
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著者には宣言する財政的競争上の利益はない。

ここに記載されているマウス肝臓裂傷モデルは、制御されない出血の信頼できる一貫したモデルを提供する。このモデルは簡単に実行できますが、細心の注意を払う必要のある重要なステップがあります。このモデルの最も技術的に困難な部分は、血行動態モニタリングおよび流体/薬物投与のための大腿血管のカニューレ挿入である。神経の切開および動脈切開/静脈切開の際には注意が必?...

外傷は、世界中の若者の死と障害の主要な原因です。 1制御されない出血は、重傷を負った外傷患者の間でも死亡原因の主要な原因のままである。出血性外傷患者の管理は、外科的出血の制御、および蘇生および失われた血液の置換の2つである。 出血性ショックの動物モデルは、外傷研究の基礎であり、病態生理学の評価および外傷性/出血性ショックの治療に使用することができる。動物モデルにおけるショックは、制御された出血および制御されない出血の2つの方法によって広く達成することができる。 5、6制御された出血は、一定量の血液を除去することによって、または一定の血圧（一定圧力）を達成するために血液を除去することによって行われる。一方、seモデルは、出血性ショックのメカニズムおよび免疫変化における評価に有用であり、止血剤の試験には適用できず、外傷後の出血の臨床シナリオを模倣しない。この程度まで、我々は、マウスモデルにおける止血変化および凝固促進剤を試験することができる制御されない出血のモデルを開発しようとした。肝臓は、肝臓への二重血液供給のために部分的に制御されない出血のための魅力的な選択肢であり、鈍的および侵襲的外傷の両方において最も傷ついた腹内側臓器の1つである。高い臨床的関連性を考慮すると、肝臓は、制御されない出血のモデルとして、最も一般的にはラットおよびブタモデルで利用されているが、最近霊長類においても同様に利用されている。 7,8,9,10,11,12 </sup&gt;マウスモデルはまた、挫傷モデルまたは鈍的外傷のような肝臓損傷を取り込んだ;しかし、これらのモデルは、肝障害に続発する出血性ショックをもたらさない。 13,14 コントロールされていない肝出血のラットおよびブタモデルは、蘇生術および血行力学的モニタリングを検討する上で貴重であるが、コスト、利用動物数、および分析に利用できる重要なトランスジェニック系統の欠如など、様々な理由によりネズミモデルよりもあまり有利ではない特定の細胞および分子シグナル伝達現在のマウスモデルは、標準化された肝臓裂傷、失血定量化、血行力学的モニタリング、および生存分析を行う能力を含む既存の肝出血モデルとの重要な類似点を共有する。多くの既存のモデルは、これらの側面のいくつかを取り入れているだけであるが、本発明者らのモデルは生理学的バリエーションの多くを測定するために開発された同時に複数のマウスに繁殖する。同様に、マウスモデルの開発は、高度な分子技術を用いたコスト効率のよいハイスループットモデルの潜在的可能性を伴って、蘇生を超えた研究および制御されない出血のより大きな病態生理学的機構への扉を開く。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>SS/45 dumonts</td>
			<td>Fine Science Tools</td>
			<td>11203-25</td>
		</tr>
		<tr>
			<td>surgical scissors</td>
			<td>Fine Science Tools</td>
			<td>14068-12</td>
		</tr>
		<tr>
			<td>hemostats</td>
			<td>Fine Science Tools</td>
			<td>13009-12</td>
		</tr>
		<tr>
			<td>microscissors</td>
			<td>Fine Science Tools</td>
			<td>15000-08</td>
		</tr>
		<tr>
			<td>0.8mm curved forceps</td>
			<td>Fine Science Tools</td>
			<td>11009-13</td>
		</tr>
		<tr>
			<td>suture reel 6-0</td>
			<td>Fine Science Tools</td>
			<td>18020-60</td>
		</tr>
		<tr>
			<td>suture 4-0 silk w/ needle</td>
			<td>Owens Minor</td>
			<td>K188H</td>
		</tr>
		<tr>
			<td>gauze 4x4</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>cotton-tip applicator</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>30G needle</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>23G needle</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>10cc syringe</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>50cc conical tube</td>
			<td colspan="2">can be purchased through any global vendor</td>
		</tr>
		<tr>
			<td>1cc syringe w/ 25G needle</td>
			<td>Fisher Scientific</td>
			<td>14-826-88</td>
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			<td>Polyethylene 10 tubing 100`(PE-10)</td>
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			<td>3-way stopcock</td>
			<td>Fisher Scientific</td>
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			<td>surgical blue pad</td>
			<td>Fisher Scientific</td>
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			<td>Sterile Field dressings</td>
			<td>Fisher Scientific</td>
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			<td>tape rolls 1&#34;</td>
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			<td>straight side wide mouth jars</td>
			<td>VWR</td>
			<td>159000-058</td>
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			<td>stainless steel tray 8&#34; x 11&#34;</td>
			<td>VWR</td>
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			<td>male-male leur lock 3-way</td>
			<td>VWR</td>
			<td>20068-909</td>
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			<td>sterilization pouch 3&#34;x8&#34;</td>
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			<td>sterilization pouch 5&#34;x10&#34;</td>
			<td>VWR</td>
			<td>24010</td>
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			<td>absorption triangles</td>
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			<td>7mm wound clip applier</td>
			<td>Fisher Scientific</td>
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			<td>1000 7mm wound clips</td>
			<td>Fisher Scientific</td>
			<td>E0522687</td>
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			<td>betadine (4oz)</td>
			<td colspan="2">can be purchased through any global vendor</td>
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			<td>sterile gloves</td>
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			<td>eppendorfs&#160;</td>
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			<td>1/2cc Lo-Dose insulin syringe</td>
			<td>Fisher Scientific</td>
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			<td>small weigh boat</td>
			<td colspan="2">can be purchased through any global vendor</td>
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			<td>lactated ringers</td>
			<td colspan="2">can be purchased through any global vendor</td>
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			<td>hepranized saline solution (.1&#181; hep + 9.9&#181;NaCl)</td>
			<td colspan="2">can be purchased through any global vendor</td>
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			<td>phosphate buffered saline&#160;</td>
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			<td>pentobarbital&#160;</td>
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			<td>Wild M650 microscope w/ boom stand</td>
			<td>Leica</td>
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			<td>Digi-Med BPA-400 analyzer &#38; systems integrator</td>
			<td>Micro-Med</td>
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			<td>TXD-310 (Digi-Med Transducer)&#160;</td>
			<td>Micro-Med</td>
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			<td>Computer</td>
			<td>Dell</td>
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			<td>microbead instrument sterilizer</td>
			<td>VWR</td>
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			<td>rectal thermometer</td>
			<td>Kent Scientific</td>
			<td>RET-3</td>
		</tr>
	</tbody>
</table>

uncontrolled hemorrhagic shock modeled via liver laceration in mice with real time hemodynamic monitoring

制御不能な出血は、外傷患者の予防可能な死亡の重要な原因である。我々は、一貫した失血、血行動態の変化、および生存をもたらす肝臓の裂傷を介した制御されない出血のマウスモデルを開発した。 マウスは肝臓の左中葉の標準化切除を受ける。彼らは機械的介入なしに出血することができます。止血剤は、治験責任医師の興味に応じて、治療前または救済療法として投与することができる。出血時に、左大腿動脈ラインを介したリアルタイムの血行動態モニタリングが行われる。次いで、マウスを屠殺し、失血を定量し、さらなる分析のために血液を採取し、臓器を傷害の分析のために採取する。実験的な設計は、複数の動物の同時試験を可能にすることが記載されている。 制御不能な出血のモデルとしての肝臓出血が存在する主にラットおよびブタのモデルにおける文献。これらのモデルの中には、血行力学的モニタリングを利用するものや失血量を定量化するものの、一貫性がないモデルもあります。現在のモデルは、コントロールされていない出血の病態生理学的機序をさらに調べるために、トランスジェニック系統およびハイスループット機構を用いる利点を提供する、ネズミモデルにおける失血のリアルタイム血行動態モニタリングの定量化を組み込んでいる。

肝臓の裂傷モデルは、マウスにおいて再現性のある一貫した失血をもたらす。 図1Aは、0.02gの標準偏差で得ることができる裂傷肝臓の一貫した重量を示す。このような刻み目のある肝臓重量の一貫性は、マウスと異なる蘇生プロトコールのような異なる実験設定においてモデルを再現する能力を可能にする。同様に、0.01gの標準誤差を有する刻んだ肝臓の?...

Watch this Scientific Journal Video about Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring at JoVE.com

Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

外傷患者の死亡原因の重要な原因である制御不能な出血は、マウスモデルにおける標準的な肝臓裂傷を用いてモデル化することができる。このモデルは一貫した失血、生存率をもたらし、止血剤の試験を可能にする。この記事では、この貴重なモデルを実行するためのステップバイステップのプロセスを提供します。

リアルタイム血行動態モニタリングを用いたマウスの肝梗塞モデルによる制御不能出血性ショック

Uncontrolled hemorrhage is an important cause of preventable deaths among trauma patients. We have developed a murine model of uncontrolled hemorrhage via ...

uncontrolled-hemorrhagic-shock-modeled-via-liver-laceration-mice-with

Research

JoVE Journal

Medicine

8.5K ビュー.  University of Pittsburgh. 外傷患者の死亡原因の重要な原因である制御不能な出血は、マウスモデルにおける標準的な肝臓裂傷を用いてモデル化することができる。このモデルは一貫した失血、生存率をもたらし、止血剤の試験を可能にする。この記事では、この貴重なモデルを実行するためのステップバイステップのプロセスを提供します。 

ビデオ: Uncontrolled Hemorrhagic Shock Modeled via Liver Laceration in Mice with Real Time Hemodynamic Monitoring

Fixed Volume or Fixed Pressure: A Murine Model of Hemorrhagic Shock

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, 2, 3, 4, 5 These signaling events can also lead to sepsis and/or multiple organ dysfunction (MOD). 6, 7, 8, 9 It is critical then to investigate the causes of suppressed immune function and detrimental signaling cascades in order to develop more effective ways to help patients who suffer from traumatic injuries. 10 This fixed pressure Hemorrhagic Shock (HS) procedure, although technically challenging, is an excellent resource for investigation of these pathophysiologic conditions. 11, 12, 13 Advances in the assessment of biological systems, i.e. Systems Biology have enabled the scientific community to further understand complex physiologic networks and cellular communication patterns. 14 Hemorrhagic Shock has proven to be a vital tool for unveiling these cellular communication patterns as they relate to immune function. 15, 16, 17, 18 This procedure can be mastered! This procedure can also be used as either a fixed volume or fixed pressure approach. We adapted this technique in the murine model to enhance research in innate and adaptive immune function. 19, 20, 21 Due to their small size HS in mice presents unique challenges. However due to the many available mouse strains, this species represents an unparalleled resource for the study of the biologic responses. The HS model is an important model for studying cellular communication patterns and the responses of systems such as hormonal and inflammatory mediator systems, and danger signals, i.e. DAMP and PAMP upregulation as it elicits distinct responses that differ from other forms of shock. 22, 23, 24, 25 The development of transgenic murine strains and the induction of biologic agents to inhibit specific signaling have presented valuable opportunities to further elucidate our understanding of the up and down regulation of signal transduction after severe blood loss, i.e. HS and trauma 26, 27, 28, 29, 30. 
There are numerous resuscitation methods (R) in association with HS and trauma. 31, 32, 33, 34 A fixed volume resuscitation method of solely lactated ringer solution (LR), equal to three times the shed blood volume, is used in this model to study endogenous mechanisms such as remote organ injury and systemic inflammation. 35, 36, 38 This method of resuscitation is proven to be effective in evaluating the effects of HS and trauma 38, 39.

The Hemorrhagic Shock model has been a reliable and reproducible resource facilitating the identification and understanding of signaling cascades associated with inflammation and end-organ damage after trauma. This article provides a step-by-step description of surgical and mechanical aspects associated with the Hemorrhagic Shock experimental procedure in mice.

固定ボリュームまたは固定圧力：出血性ショックのマウスモデル

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, ...

医学

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

ヒト全血でのリアルタイム細胞毒性アッセイ

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory&#160;rate&#160;and heart&#160;rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.
This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41&#177;2.94 cmH2O which was significantly higher (P&#60;0.05) than before resection (6.87&#177;2.39 cmH2O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.

This film demonstrates how to acquire systemic and hepatic hemodynamics in mice. The whole monitoring includes acquisition of vital parameters, systemic blood pressure, central venous pressure, common hepatic artery flow rate, and portal vein pressure as well as the portal flow rate in mice.

マウスにおける全身および肝血行動態パラメータの監視

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

マウスにおける急性心筋梗塞のリアルタイム圧解析

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the forefront of trauma care in recent years. Since complete aortic occlusion presents serious concerns, the concept of partial aortic occlusion has gained a growing attention. Here, we present a large animal model of hemorrhagic shock to investigate the effects of a novel partial aortic balloon occlusion catheter and compare it with a catheter that works on the principles of complete aortic occlusion. Swine are anesthetized and instrumented in order to conduct controlled fixed-volume hemorrhage, and hemodynamic and physiological parameters are monitored. Following hemorrhage, aortic balloon occlusion catheters are inserted and inflated in the supraceliac aorta for 60 min, during which the animals receive whole-blood resuscitation as 20% of the total blood volume (TBV). Following balloon deflation, the animals are monitored in a critical care setting for 4 h, during which they receive fluid resuscitation and vasopressors as needed. The partial aortic balloon occlusion demonstrated improved distal mean arterial pressures (MAPs) during the balloon inflation, decreased markers of ischemia, and decreased fluid resuscitation and vasopressor use. As swine physiology and homeostatic responses following hemorrhage have been well-documented and are like those in humans, a swine hemorrhagic shock model can be used to test various treatment strategies. In addition to treating hemorrhage, aortic balloon occlusion catheters have become popular for their role in cardiac arrest, cardiac and vascular surgery, and other high-risk elective surgical procedures.

Here, we present a protocol demonstrating a hemorrhagic shock model in swine that uses aortic occlusion as a bridge to definitive care in trauma. This model has application in testing a wide range of surgical and pharmacological therapeutic strategies.

豚の出血性ショックの治療のための完全なおよび部分的な大動脈閉塞症

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the ...

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

肝静脈圧勾配と経頸部肝生検の測定

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

Standardized Hemorrhagic Shock Induction Guided by Cerebral Oximetry and Extended Hemodynamic Monitoring in Pigs

Hemorrhagic shock ranks among the main reasons for severe injury-related death. The loss of circulatory volume and oxygen carriers can lead to an insufficient oxygen supply and irreversible organ failure. The brain exerts only limited compensation capacities and is particularly at high risk of severe hypoxic damage.This article demonstrates the reproducible induction of life-threatening hemorrhagic shock in a porcine model by means of calculated blood withdrawal. We titrate shock induction guided by near-infrared spectroscopy&#160;and extended hemodynamic monitoring to display systemic circulatory failure, as well as cerebral microcirculatory oxygen depletion. In comparison to similar models that primarily focus on predefined removal volumes for shock induction, this approach highlights a titration by means of the resulting failure of macro- and microcirculation.

Hemorrhagic shock is a severe complication in seriously injured patients, which leads to life-threatening oxygen undersupply. We present a standardized method to induce hemorrhagic shock via blood withdrawal in pigs that is guided by hemodynamics and microcirculatory cerebral oxygenation.

ブタの脳オキシメトリーと拡張血行力モニタリングによって導かれる標準化された出血性ショック誘導

Hemorrhagic shock ranks among the main reasons for severe injury-related death. The loss of circulatory volume and oxygen carriers can lead to an ...

Real-Time Monitoring of Neurocritical Patients with Diffuse Optical Spectroscopies

Neurophysiological monitoring is an important goal in the treatment of neurocritical patients, as it may prevent secondary damage and directly impact morbidity and mortality rates. However, there is currently a lack of suitable non-invasive, real-time technologies for continuous monitoring of cerebral physiology at the bedside. Diffuse optical techniques have been proposed as a potential tool for bedside measurements of cerebral blood flow and cerebral oxygenation in case of neurocritical patients. Diffuse optical spectroscopies have been previously explored to monitor patients in several clinical scenarios ranging from neonatal monitoring to cerebrovascular interventions in adults. However, the feasibility of the technique to aid clinicians by providing real-time information at the bedside remains largely unaddressed. Here, we report the translation of a diffuse optical system for continuous real-time monitoring of cerebral blood flow, cerebral oxygenation, and cerebral oxygen metabolism during intensive care. The real-time feature of the instrument could enable treatment strategies based on patient-specific cerebral physiology rather than relying on surrogate metrics, such as arterial blood pressure. By providing real-time information on the cerebral circulation at different time scales with relatively cheap and portable instrumentation, this approach may be especially useful in low-budget hospitals, in remote areas and for&#160;monitoring in open fields (e.g., defense and sports).

Presented here is a protocol for non-invasively monitoring cerebral hemodynamics of neurocritical patients in real-time and at the bedside using diffuse optics. Specifically, the proposed protocol uses a hybrid diffuse optical systems to detect and display real-time information on cerebral oxygenation, cerebral blood flow and cerebral metabolism.

びまん性光学分光法による神経クリティカル患者のリアルタイムモニタリング

Neurophysiological monitoring is an important goal in the treatment of neurocritical patients, as it may prevent secondary damage and directly impact ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

ラット肝移植モデルにおける血球力学的変化をシミュレートすることによる臓器虚血再灌流損傷

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...