这项工作得到了吉林省科学技术厅（YDZJ202201ZYTS181）的支持。

该动物程序已获得长春中医药大学伦理委员会批准（备案号:2021214）。将无特定病原体的C57BL / 6J小鼠（8-10周，体重18-22g），雄性和雌性，饲养在22°C和65%相对湿度的独立通风笼中。小鼠在适应性喂养7天后开始实验，在此期间它们可以自由获得水和饮食。
1.药物制备
<ol><li>LJZD的制备 注:所用中药购自长春中医药大学附属医院，经鉴定为正品中药（见 表1）。<ol><li>将当神（12g）、白术（12g）、赣草（6g）、陈皮（12g）、姜板峡（9g）放入专用陶瓷锅中（见 材料表）。加入1000mL蒸馏水，在室温下浸泡1小时（见 图1A）。</li>
		<li>粉末12g涪凌用研磨机研磨成细粉，并在室温下在另一个容器中的300mL蒸馏水中浸泡1小时。</li>
		<li>将上述中药混合在专用陶瓷锅中。将混合物煮沸，然后保持中火，直到只剩下 300 mL 汤剂。使用医用纱布进行过滤，并将滤液保存在室温下。</li>
		<li>加入 1000 mL 蒸馏水，再次重复上述煎煮操作。再次用医用纱布过滤。将滤液混合并煮沸至仅剩 150 mL。</li>
		<li>将浓缩液体以10,000× g 离心5分钟，并将获得的上清液进一步浓缩至30mL。将最终浓缩物转移到培养皿中，并使用电动烘箱干燥，直到只有溶质保留为粉末。</li>
		<li>称取上述固体，溶于无菌蒸馏水中，得到每0.2mL（114.16mg/mL）含22.85mg药物的溶液，即为小鼠日用量。</li>
	</ol></li>
	<li>5-氨基水杨酸的制备 注:5-氨基水杨酸（5-ASA;见 材料表）对UC-CRC有良好的预防作用，在本研究中被用作阳性药物23。<ol><li>将 64 mg 5-ASA 粉末溶解在 200 mL 无菌蒸馏水中，得到 1.82 mg/mL 5-ASA 溶液。单只小鼠的日剂量为0.2mL。</li>
	</ol></li>
	<li>AOM注射液的制备<ol><li>将2.5mL无菌蒸馏水加入25mg AOM粉末中（见 材料表），并通过涡旋混合器（见 材料表）混合制成10mg / mL AOM储备溶液，储存在-20°C直至使用。</li>
		<li>通过用无菌蒸馏水以 10:1 （1 mg/mL） 稀释 AOM 储备溶液来制备 AOM 注射液。</li>
	</ol></li>
</ol>2. UC-CRC模型的建立
注:实验分为4组:对照组、模型组、LJZD组和5-ASA组，每组10只小鼠。除对照组外，其他组均接受AOM和DSS治疗。
<ol><li>腹腔注射AOM注射液 注意:适应性喂养7天后，通过腹膜内注射给予小鼠AOM注射液（1mg / mL）（见 图1B）。<ol><li>保持鼠标的腹部朝上，头部略微向下。握住背部皮肤以收紧腹部皮肤，并用 1 mL 注射器刺穿中腹部线上两条大腿根线交叉处约 1 厘米处的皮肤（见 材料表）。</li>
		<li>将 1 mL 注射器针头推到皮肤下 3-5 mm 的距离，使其与腹部中线平行，然后将针头以 45° 插入腹腔 0.3-0.5 mm。</li>
		<li>尖端穿过腹部肌肉后，操作者会感觉到阻力突然丧失。随后，向外和向后拉动注射器，观察是否发生液体渗漏。如果没有，将AOM注射液以0.1mL / 10g缓慢推入小鼠体内。</li>
	</ol></li>
	<li>2%DSS溶液的循环刺激 注意:每只AOM诱导的小鼠在第3、6和9周给予500mL DSS溶液，在此期间小鼠自由饮水。<ol><li>通过向 10 g DSS 中加入 500 mL 无菌蒸馏水来制备 2% DSS 溶液（参见 材料表）。与涡旋混合器混合，并在4°C下储存直至使用。</li>
		<li>在AOM诱导后的第3周，第6周和第9周，每个AOM诱导的小鼠自由饮用500mL 2%DSS溶液7天。</li>
	</ol></li>
</ol>3.药物治疗
注意:成年人每天需要 63 克 LJZD。根据实验小鼠与人药物剂量的换算公式，小鼠的当量实验剂量（mg/kg）=人剂量（mg/kg）/体重（60kg）×9.1，小鼠的日剂量约为9.6g/kg。
<ol><li>分别在第 7 周和第 15 周通过胃管饲法用 0.1 mL/10 g 制备的 LJZD 和 5-ASA 溶液治疗 LJZD 和 5-ASA 组。<ol><li>对于小鼠的胃内给药，请执行以下程序。左手握住鼠标，右手握住胃灌注装置。将注射器针头插入口腔，然后将其滑下小鼠咽部的后壁。当小鼠吞咽时，沿着咽部滑动并继续前进。当有抵抗感，注射器可以推入咽部时，拔出针头并完成注射。</li>
	</ol></li>
	<li>用相同量的生理盐水处理对照组和模型组（参见 材料表）。</li>
	<li>在给药期间每天一次用适当的药物治疗每组小鼠。</li>
</ol>4. UC-CRC模型的评价及LJZD的疗效
<ol><li>疾病活动指数评分 注:根据 表2，通过结合小鼠的体重减轻，粪便粘度和粪便出血来评估疾病活动指数（DAI）评分。<ol><li>从适应性喂养开始到药物治疗结束，每天记录小鼠的体重。</li>
		<li>仔细观察每只实验小鼠的粪便稠度，并将排便记录为三种情况之一:正常、稀便和水样腹泻。</li>
		<li>将实验动物的粪便出血记录为三种情况之一:无出血、少量出血、便血可见。</li>
	</ol></li>
	<li>检测血清中IL-6水平<ol><li>用LJZD或5-ASA治疗小鼠9周。用左手抓住小鼠的颈部皮肤，轻轻按压实验台，采取侧卧位，固定小鼠。用剪刀剪掉老鼠的胡须。用剪刀小心地剪掉小鼠的胡须（见 材料表），以防止血液污染。 注意:不能自由摆动的老鼠被认为是正确固定的。如果没有发生这种情况，则应固定鼠标。</li>
		<li>通过吸入2%异氟烷麻醉小鼠。用乙醇对眼球周围的皮肤进行消毒（见 材料表）。轻轻按压侧面的眼部皮肤，使眼球充血突出。</li>
		<li>用肘部镊子夹住眼球，准确快速地取出眼球。让血液滴入离心管中（见 材料表）。在此过程中，点击鼠标的心脏以加快采血速度。</li>
		<li>将收集的血液在室温下保持30分钟，并以3,500× g 离心10分钟。收集上清液并根据IL-6含量检测试剂盒说明检测IL-6水平（参见 材料表）。</li>
	</ol></li>
	<li>分离结直肠组织<ol><li>根据动物伦理，通过吸入5%过量的异氟醚和宫颈脱位（见 材料表）对小鼠进行血液采样后实施安乐死。</li>
		<li>将小鼠保持在低温解剖环境中。将小鼠固定在仰卧位。用剪刀剪下腹部毛发，并用乙醇消毒。</li>
		<li>用眼睑镊夹住两个大腿根部和腹部中线之间的交点（见 材料表）。用剪刀剪一个约1-1.5厘米的横切口。</li>
		<li>从横切口的中点沿腹部正中线向剑突夹一个纵向切口。</li>
		<li>沿肛门方向取出结直肠周围组织，将结直肠与周围组织分开。注意不要损坏结直肠。</li>
		<li>将腹部皮肤向两侧推，使结直肠完全暴露。用眼睑镊子从腹腔中取出结直肠，并切断从肛门到盲肠的部分（不包括）;总长度约为10厘米。将获得的结直肠组织储存在4°C的盐水中。</li>
	</ol></li>
	<li>直肠长度和重量的评估<ol><li>用5mL针头在4°C下提取生理盐水（参见 材料表）以冲洗结直肠内部。然后，将结直肠放在吸水纸上以吸收组织的水分。</li>
		<li>称量结直肠组织，然后将它们放在 A4 纸上测量其长度。</li>
	</ol></li>
	<li>测量结直肠肿瘤的数量<ol><li>纵向切开结直肠以使其完全展开，并观察结直肠中肿瘤的数量和大小。</li>
	</ol></li>
	<li>结直肠的病理分析<ol><li>将结直肠固定在4%多聚甲醛中24小时。将固定的直肠组织嵌入熔化的石蜡中，并通过组织冷冻切片机以5μm的厚度连续切片。</li>
		<li>根据Hou等人24的程序，在二甲苯中对切片进行脱蜡，然后用连续浓度的乙醇脱水。用苏木精溶液染色5分钟后，用纯净水冲洗切片。之后，用0.5%曙红溶液染色1分钟（见 材料表）。</li>
		<li>再次进行梯度脱水和二甲苯透明处理。密封切片并在光学显微镜下观察它们（见 材料表）和照片，如Xie等人25所述。</li>
	</ol></li>
	<li>结直肠的免疫组织化学分析<ol><li>按上述方法对切片进行脱蜡和脱水。用高压热修复技术修复切片中的抗原，如Gok等人26所述。</li>
		<li>将切片在室温下浸泡在内源性过氧化物酶阻滞剂中15分钟。用山羊血清密封切片（见 材料表）。</li>
		<li>将一抗ZO-1（1:1000），Occludin（1:1000）和KI67（1:500;参见 材料表）加入切片中，并在4°C下孵育过夜。 用PBS缓冲液冲洗切片（见 材料表），加入通用二抗（1:5000;见 材料表），并在37°C孵育30分钟。</li>
		<li>将 DAB 解决方案（请参阅 材料表）添加到显色部分。用苏木精溶液对切片进行复染。</li>
		<li>再次对切片进行脱水、透明和密封。通过光学显微镜观察蛋白质的表达。</li>
	</ol></li>
</ol>

六君子汤:预防 UC-CRC 的有效治疗方法

作者没有什么可透露的。

结直肠癌是全球最常见的癌症之一，每年约有 1,148,000 例新发病例和超过 576,000 例死亡。结直肠癌根据病因不同可分为3种类型，包括遗传性、散发性和UC-CRC31。UC等炎症性肠病患者的CRC发病率明显高于一般人群。UC 通过炎症-癌症通路刺激 CRC 的发展，这与典型的腺瘤-腺癌通路不同 6。目前UC-CRC的病因不明，主要由长期复发性慢性炎症引起，死亡率高达60%<sup class="...

结直肠癌（CRC）是一种常见的胃肠道恶性肿瘤，是全球第三大常见恶性肿瘤，也是全球第二大常见死因，占全球癌症发病率的10%，占癌症相关死亡总数的9.4%1,2。遗传因素、慢性炎症、高脂肪饮食、糖尿病和肠道菌群异常是 CRC 3,4 的危险因素。其中，炎症性肠病，尤其是溃疡性结肠炎（UC），是CRC 5,6的明显危险因素。UC相关结直肠癌（UC-CRC）是基于结直肠慢性炎症的炎症、非典型增生和癌症的过渡过程，与CRC 7,8的典型腺瘤-腺癌发展模型不同。与一般人群相比，炎症性肠病患者患CRC的风险大约高出10-40倍9。
目前，手术仍然是 CRC 的标准治疗方法，并且根据肿瘤的位置和阶段，放射治疗、全身药物治疗或两者的组合是可能的10。虽然这些传统的治疗方式已经取得了很大的进展，但由于结直肠癌的异质性和复发率高，预后较差，治疗效果不理想11,12。因此，早期发现、早期诊断、综合治疗是提高结直肠癌患者生存率的关键，关注结直肠癌向结直肠癌的转化尤为重要。多年来，中医药因其副作用有限、疗效显著而在治疗UC-CRC或慢性胃炎方面备受关注。历代著名中医在辩证法的基础上，创作了大量的经典药方，如黄旗建中汤13、四君子汤14、思神丸15等。
流君子汤（LJZD）起源于明代编纂的易雪正传著作，是中医16中的经典处方。如表1所示，LJZD由六种传统草本植物组成，包括党参（Codonopsis pilosula，Franch.）南夫。（当神）、茯苓（Schw.）、狼（涪陵）、苍术（Atractylodes macrocephala Koidz）。（白术）， Glycyrrhiza uralensis Fisch.（Gancao）、柑橘（Citrus reticulata Blanco）和Pinellia ternata（Thunb.）蒲公英（半夏），具有补气健脾、干湿化痰的作用。在现代临床实践中，常用于治疗慢性胃炎、胃溃疡和十二指肠溃疡。现代药理学研究表明，LJZD和改良的LJZD在UC和消化道癌的辅助治疗中具有较高的应用价值17,18,19。
目前UC-CRC小鼠模型的构建方法很多，但偶氮甲烷（AOM）/葡聚糖硫酸钠（DSS）诱导的小鼠模型是应用最广泛的UC-CRC模型;临床症状、形态学和病理学观察证明，该模型与人类UC-CRC20,21非常相似。其基本原理是先用化学致癌物AOM诱导致癌作用，然后连续将小鼠暴露在DSS的炎症刺激环境中，模拟肠粘膜上皮的连续损伤和修复，从而构建UC-CRC小鼠模型22。本研究的目的是在短期内通过腹腔注射AOM和DSS的循环刺激建立UC-CRC的小鼠模型，并评价该药物和LJZD对UC-CRC的分子机制，以期为UC-CRC的治疗提供科学依据。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Azoxymethane</td>
			<td>Sigma</td>
			<td>A5486</td>
			<td></td>
		</tr>
		<tr>
			<td>5-amino salicylic acid</td>
			<td>Kuihua Pharmaceuticals Group Jiamusi Luling Pharmaceutical Co., Ltd</td>
			<td>3819413</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6J mice</td>
			<td>Liaoning Changsheng Biotechnology Co., Ltd</td>
			<td>NO 210726210100853716</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover slip</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>10212432C</td>
			<td></td>
		</tr>
		<tr>
			<td>DAB color development kit</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>2005289</td>
			<td></td>
		</tr>
		<tr>
			<td>Dewatering machine&#160;</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JJ-12J</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextran sulfate sodium</td>
			<td>Dalian Meilun Biotechnology Co., Ltd</td>
			<td>MB5535</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding machine</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-P5</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin-eosin dye</td>
			<td>Wuhan Hundred Degree Biotechnology Co., Ltd</td>
			<td>B1000</td>
			<td></td>
		</tr>
		<tr>
			<td>IL-6</td>
			<td>Jiangsu Meimian Industrial Co., Ltd</td>
			<td>MM-0163M2</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science Co., Ltd</td>
			<td>R510-22-10</td>
			<td></td>
		</tr>
		<tr>
			<td>KI67 primary antibody</td>
			<td>Google Biotechnology Inc</td>
			<td>GB121141</td>
			<td></td>
		</tr>
		<tr>
			<td>Neutral gum</td>
			<td>Wuhan Hundred Degree Biotechnology Co., Ltd</td>
			<td>10004160</td>
			<td></td>
		</tr>
		<tr>
			<td>Object slide</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>10212432A</td>
			<td></td>
		</tr>
		<tr>
			<td>Occludin primary antibody</td>
			<td>Affnity</td>
			<td>DF7504</td>
			<td></td>
		</tr>
		<tr>
			<td>Orthostatic optical microscope</td>
			<td>Nikon</td>
			<td>Nikon Eclipse CI</td>
			<td></td>
		</tr>
		<tr>
			<td>Pathological microtome</td>
			<td>Shanghai Leica Instrument Co., Ltd</td>
			<td>RM2016</td>
			<td></td>
		</tr>
		<tr>
			<td>ZO-1 primary antibody</td>
			<td>Abcam</td>
			<td>ab221547</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishment of coloproctitis cancer model in mice and evaluation of therapeutic effect of chinese medicine

结直肠癌 （CRC） 是一种常见的消化系统恶性肿瘤，已成为全球第三大常见恶性肿瘤和恶性肿瘤相关死亡的第二大原因。溃疡性结肠直肠炎 （UC） 是一种癌前病变，UC 相关 CRC （UC-CRC） 是 CRC 最常见的亚型。因此，合理的UC-CRC模型是新药研发的基石和保障。中医药（TCM）因其良好的疗效而被广泛用于治疗UC-CRC。柳君子汤（LJZD）作为中医的经典补药方，已被广泛用于治疗UC-CRC。本研究结合偶氮甲烷和硫酸葡聚糖钠建立UC-CRC模型，并给予LJZD。数据证实，LJZD可利用小鼠体重、结直肠长度、病理炎症因子、结直肠屏障功能、癌症标志物等手段有效抑制UC-CRC的癌症转化。该协议提供了一个评估中医预防和治疗UC-CRC疗效的系统。

LJZD的汤剂按表1中药物的组成比例和图1A中的中药的汤剂方法制备。根据图1B所示的时间点，小鼠在第7天腹膜内注射1 mg/mL AOM，在第3、6、9周自由饮用含有2%DSS的饮用水。UC-CRC小鼠模型在第15周成功建立。同时，从第7周到第15周，通过强饲法对小鼠进行LJZD处理。数据显示，与对照组相比，UC-CRC模型组?...

Watch this Scientific Journal Video about Author Spotlight: Liujunzi Decoction as a Traditional Chinese Treatment for Coloproctitis Cancer at JoVE.com

作者聚焦: 六君子汤作为结肠直肠炎癌症的中医治疗方法

该方案提供了由偶氮甲烷与硫酸葡聚糖钠联合诱导的溃疡性结肠直肠炎相关结直肠癌的小鼠模型。该模型用于评价中药化合物在预防和治疗结直肠癌中的疗效。

小鼠结肠直肠炎癌症模型的建立及中药治疗效果评价

Colorectal cancer (CRC) is a common malignancy of the digestive system and has become the third most common malignancy worldwide and the second leading ...

这项研究是关于用中医预防和治疗结肠直肠炎癌症的。我们旨在建立结肠直肠炎癌症的小鼠模型并评估中医指标。我们证实了中药流君子汤是预防结肠直肠炎癌症的有效方法。未来，我们将致力于识别结肠癌和直肠癌更敏感的生物标志物，并筛选流君子汤的治疗靶点。

establishment-coloproctitis-cancer-model-mice-evaluation-therapeutic

Research

JoVE Journal

Medicine

Establishment of Coloproctitis Cancer Model in Mice and Evaluation of Therapeutic Effect of Chinese Medicine

734 次观看.  Changchun University of Chinese Medicine. 这项研究是关于用中医预防和治疗结肠直肠炎癌症的。我们旨在建立结肠直肠炎癌症的小鼠模型并评估中医指标。我们证实了中药流君子汤是预防结肠直肠炎癌症的有效方法。未来，我们将致力于识别结肠癌和直肠癌更敏感的生物标志物，并筛选流君子汤的治疗靶点。

视频: 作者聚焦: 六君子汤作为结肠直肠炎癌症的中医治疗方法

Establishment and Propagation of Human Retinoblastoma Tumors in Immune Deficient Mice

Culturing retinoblastoma tumor cells in defined stem cell media gives rise to primary tumorspheres that can be grown and maintained for only a limited time. These cultured tumorspheres may exhibit markedly different cellular phenotypes when compared to the original tumors. Demonstration that cultured cells have the capability of forming new tumors is important to ensure that cultured cells model the biology of the original tumor. 
Here we present a protocol for propagating human retinoblastoma tumors in vivo using Rag2-/- immune deficient mice. Cultured human retinoblastoma tumorspheres of low passage or cells obtained from freshly harvested human retinoblastoma tumors injected directly into the vitreous cavity of murine eyes form tumors within 2-4 weeks. These tumors can be harvested and either further passaged into murine eyes in vivo or grown as tumorspheres in vitro. Propagation has been successfully carried out for at least three passages thus establishing a continuing source of human retinoblastoma tissue for further experimentation. 
Wesley S. Bond and Lalita Wadhwa are co-first authors.

A method is described to propagate human retinoblastoma tumors in mice. Tumor cells are directly injected into the eyes of immune deficient mice. Secondary tumors have been successfully established using both cells directly harvested from human tumors and cultured tumorspheres.

在免疫缺陷小鼠的建立和传播人类视网膜母细胞瘤肿瘤

Culturing retinoblastoma tumor cells in defined stem cell media gives rise to primary tumorspheres that can be grown and maintained for only a limited ...

科研

医学

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold standard treatment for the high-grade non-muscle invasive bladder cancer (NMIBC). However, since the treatment-related side effects are relevant, newer biological response modifiers with a better benefit/side effects ratio are needed.
The tumour microenvironment can influence both tumour development and therapy efficacy. In order to obtain a good model, it is desirable to implant tumour cells in the organ from which the cancer originates.
In this protocol, we describe a method for establishing a tumour in the bladder cavity of female mice and subsequent delivery of therapeutic agents; the latter are exemplified by our use of Helicobacter pylori neutrophil activating protein (HP-NAP). A preliminary chemical burn of the mucosa, followed by the injection of mouse urothelial carcinoma cell line MB49 via urethral catheterization, enables the cells to attach to the bladder mucosa. After a period, required to allow an initial proliferation of the cells, mice are treated with HP-NAP, administrated again via catheterization. The anti-tumour activity of HP-NAP is evaluated comparing the tumour volume, the extent of necrosis and the degree of vascularization between vehicle- and HP-NAP-treated animals.

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Here the method to establish a syngeneic mouse model of orthotopic bladder tumour to evaluate the anti-tumour efficacy of the bacterial protein HP-NAP is described.

评价疗效<em&gt;小时。幽门螺杆菌</em&gt;蛋白HP-NAP为膀胱癌的治疗在原位小鼠模型的治疗工具

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...

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.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

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

Establishment and Evaluation of a Porcine Vein Graft Disease Model

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the investigation of disease mechanisms and the development of therapeutic strategies. 
To perform the surgery, we enter the cardiac chamber through the third intercostal space and carefully dissect the internal mammary vein and immerse it in normal saline. The right main coronary artery is then treated for ischemia. The target vessel is incised, a shunt plug is placed, and the distal end of the graft vein is anastomosed. The ascending aorta is partially blocked, and the proximal end of the graft vein is anastomosed after perforation. The graft vein is checked for patency, and the proximal right coronary artery is ligated. 
CABG surgery is performed in minipigs to harvest the left internal mammary vein for its use as a vascular graft. Serum biochemical tests are used to evaluate the physiological status of the animals after surgery. Ultrasound examination shows that the proximal, middle, and distal end of the graft vessel are unobstructed. In the surgical model, turbulent blood flow in the graft is observed upon histological examination after the CABG surgery, and venous graft stenosis associated with intimal hyperplasia is observed in the graft. The study here provides detailed surgical procedures for the establishment of a repeatable CABG-induced VGD model.

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.

猪静脉移植病模型的建立与评价

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the ...

Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is similar in color to the thyroid gland. The lack of effective animal models in preclinical research is a severe limitation for the development of PG identification techniques. This protocol allows for the establishment of a simple and effective rat model for PG identification. In this model, black iron oxide nanoparticles (IONPs) are injected locally in the thyroid gland and rapidly diffuse within the thyroid gland but not the PG. A negatively stained PG and a positively stained thyroid gland can be easily identified by the naked eye without requiring external microscopes. The position of the PG can be identified by increasing the color contrast between the thyroid gland and the PG, based on the color of the black IONPs. This rat model is low-cost and convenient for PG identification, and the IONPs are a novel PG contrast agent.

To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent.

建立简单有效的术中甲状旁腺成像大鼠模型

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is ...

Development and Evaluation of a Rat Model of Full-Thickness Cartilage Defects

Cartilage defects of the knee joint caused by trauma are a common sports joint injury in the clinic, and these defects result in joint pain, impaired movement, and eventually, knee osteoarthritis (kOA). However, there is little effective treatment for cartilage defects or even kOA. Animal models are important for developing therapeutic drugs, but the existing models for cartilage defects are unsatisfactory. This work established a full-thickness cartilage defects (FTCD) model by drilling holes in the femoral trochlear groove of rats, and the subsequent pain behavior and histopathological changes were used as readout experiments. After surgery, the mechanical withdrawal threshold was decreased, chondrocytes at the injured site were lost, matrix metalloproteinase MMP13 expression was increased, and type II collagen expression decreased, consistent with the pathological changes observed in human cartilage defects. This methodology is easy and simple to perform and enables gross observation immediately after the injury. Furthermore, this model can successfully mimic clinical cartilage defects, thus providing a platform for studying the pathological process of cartilage defects and developing corresponding therapeutic drugs.

This protocol establishes a full-thickness cartilage defects (FTCD) model by drilling holes in the femoral trochlear groove of rats and measuring the subsequent pain behavior and histopathological changes.

全层软骨缺损大鼠模型的开发与评价

Cartilage defects of the knee joint caused by trauma are a common sports joint injury in the clinic, and these defects result in joint pain, impaired ...

改进大鼠模型中的 FSN 治疗:最大限度地减少不适并最大限度地提高效率

Efficacy of Fu's Subcutaneous Needling on Sciatic Nerve Pain: Behavioral and Electrophysiological Changes in a Chronic Constriction Injury Rat Model

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the mechanisms of action are still not fully understood. During FSN treatment, the FSN needle is inserted and retained in the subcutaneous tissues for a long duration with a swaying movement. However, challenges arise from maintaining a posture while manipulating FSN in animal models (e.g., rats) for researchers. Uncomfortable treatment can lead to fear and resistance to FSN needles, increasing the risk of injury and may even affect research data. Anesthesia may also affect the study results too. Hence, there is a need for strategies in FSN therapy on animals that minimize injury during the intervention. This study employs a chronic constriction injury model in Sprague-Dawley rats to induce neuropathic pain. This model replicates the pain induced by nerve injury observed in humans through surgical constriction of a peripheral nerve, mimicking the compression or entrapment seen in conditions such as nerve compression syndromes and peripheral neuropathies. We introduce an appropriate manipulation for easily inserting an FSN needle into the subcutaneous layer of the animal&#39;s body, including needle insertion and direction, needle retention, and swaying movement. Minimizing the rat&#39;s discomfort prevents the rat from being tense, which causes the muscle to contract and hinder the entry of the needle and improves the study efficiency.

作者聚焦:揭示 FSN 治疗的治疗效果 – 桥接神经性疼痛的研究和临床应用

We present a protocol for using Fu's subcutaneous needling in a chronic constriction injury model to induce sciatic nerve pain in rats.

傅氏皮下针刺治疗坐骨神经疼痛的疗效:慢性缩窄损伤大鼠模型中的行为和电生理变化

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the ...

大鼠 DRG 压缩模型的标准化 Tuina 协议

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

作者聚焦： 推拿对压迫背根神经节疼痛的大鼠模型的镇痛作用  

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

推拿对大鼠模型的镇痛作用，压迫背根神经节疼痛

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...