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。

Colorectal cancer (CRC) is a common gastrointestinal malignancy, the third most common malignancy, and the second most common cause of death in the world, accounting for 10% of the global cancer incidence and 9.4% of the total cancer-related death1,2. Genetic factors, chronic inflammation, high-fat diet, diabetes, and abnormal intestinal flora are risk factors for CRC3,4. Among them, inflammatory bowel disease, especially ulcerative coloproctitis (UC), is a clear risk factor for CRC5,6. UC-associated CRC (UC-CRC) is a transition process of inflammation, atypical hyperplasia, and cancer based on chronic inflammation of the colorectum, which is different from the typical adenoma-adenocarcinoma development model of CRC7,8. Compared with the general population, the risk of CRC is approximately 10-40 times higher in patients with inflammatory bowel disease9.
Currently, surgery is still the standard treatment for CRC, and depending on the location and stage of the tumor, radiation therapy, systemic drug therapy, or a combination of both are possible10. Although these traditional treatment modalities have made great progress, due to the high heterogeneity and recurrence rate of CRC, the prognosis is poor, and the treatment effect is not ideal11,12. Therefore, early detection, early diagnosis, and comprehensive treatment are key to improving the survival rate of CRC patients, and it is particularly important to pay attention to the transformation of UC to CRC. Over the years, traditional Chinese medicine (TCM) has attracted much attention in the treatment of UC-CRC or chronic gastritis due to its limited side effects and significant efficacy. Based on dialectical treatment, famous Chinese medicine practitioners of various generations have created a large number of classic prescriptions, such as Huangqi Jianzhong decoction13, Sijunzi decoction14, and Sishen pill15.
Liujunzi decoction (LJZD) originated from the works of Yi Xue Zheng Zhuan compiled in the Ming Dynasty and is a classic prescription in TCM16. As shown in Table 1, LJZD consists of six traditional herbs, including Codonopsis pilosula (Franch.) Nannf. (Dangshen), Poria cocos (Schw.) Wolf (Fuling), Atractylodes macrocephala Koidz. (Baizhu), Glycyrrhiza uralensis Fisch. (Gancao), Citrus reticulata Blanco (Chenpi) and Pinellia ternata (Thunb.) Breit (Banxia), which has the effect of replenishing qi and strengthening the spleen, drying dampness, and resolving phlegm. In modern clinical practice, it is often used to treat chronic gastritis, gastric ulcers, and duodenal ulcers. Modern pharmacological research has shown that LJZD and modified LJZD have high application value in the adjuvant treatment of UC and digestive tract cancer17,18,19.
At present, there are many ways to construct UC-CRC mouse models, but the azoxymethane (AOM)/dextran sulfate sodium (DSS) induced mouse model is the most widely used UC-CRC model; the clinical symptoms, morphological, and pathological observations have proved that the model is very similar to human UC-CRC20,21. The basic principle is to first induce carcinogenesis with chemical carcinogen AOM and then continuously expose mice to the inflammatory stimulation environment of DSS to simulate the continuous damage and repair of intestinal mucosal epithelium, thereby constructing a UC-CRC mouse model22. The aim of this study is to establish a mouse model of UC-CRC by intraperitoneal injection of AOM and cyclic stimulation of DSS in the short term and to evaluate the effect of the drug and the molecular mechanism of LJZD on UC-CRC in order to provide a scientific basis for the treatment of UC-CRC.

作者聚焦： 六君子汤作为结肠直肠炎癌症的中医治疗方法

小鼠结肠直肠炎癌症模型的建立及中药治疗效果评价

This study was on the prevention and treatment of coloproctitis cancer with traditional Chinese medicine. We aim to establish a mouse model for coloproctitis cancer and evaluate traditional Chinese medicine indicators. We confirmed that the Chinese medicine Liujunzi decoction is a effective treatment for coloproctitis cancer prevention.In the future, we will work to identify more sensitive biomarkers for colon and rectal cancer and screen Liujunzi decoction for therapeutic targets.

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

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。

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

596 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

Colorectal cancer (CRC) is a common malignancy of the digestive system and has become the third most common malignancy worldwide and the second leading cause of malignancy-related death. Ulcerative coloproctitis (UC) is a precancerous lesion, and UC-associated CRC (UC-CRC) is the most common subtype of CRC. Therefore, a reasonable UC-CRC model is the cornerstone and guarantee of new drug development. Traditional Chinese medicine (TCM) has been widely used in the treatment of UC-CRC due to its good efficacy. As a classic tonic prescription of TCM, Liujunzi decoction (LJZD) has been widely used in the treatment of UC-CRC. In this study, a UC-CRC model was established by combining azomethane and dextran sulfate sodium, and the LJZD was administered. The data confirmed that LJZD can effectively inhibit cancer transition in UC-CRC by using mouse body weight, colorectal length, pathological and inflammatory factors, colorectal barrier function, and cancer markers. This protocol provides a system for evaluating the efficacy of TCM in the prevention and treatment of UC-CRC.

This protocol provides a mouse model of ulcerative coloproctitis-associated colorectal cancer induced by azomethane combined with dextran sulfate sodium. The model was used to evaluate the efficacy of traditional Chinese medicine compounds in the prevention and treatment of colorectal cancer.

科研

医学

Preparation of Liujunzi Decoction (LJZD) for the Treatment of Ulcerative Coloproctitis-Colorectal Cancer  (UC-CRC)

To begin place Dang Shen, Bai Zhu, into a specialized ceramic pot. Add 1000 milliliters of distilled water and let it soak at room temperature for one hour. Using a grinder, grind 12 grams of Fu ling into a fine powder.Then, soak the powdered Fu ling in 300 milliliters of distilled water in a separate container for one hour at room temperature. Mix the prepared Chinese herbal mixture in the specialized ceramic pot. Boil the mixture and maintain it over medium heat until the decoction volume reduces to 300 milliliters.Combine the filtrate obtained from the decoction and continue boiling until the volume reduces to 150 milliliters. Then centrifuge the concentrated liquid at 10, 000 G for five minutes. Concentrate the obtained supernatant to 30 milliliters over medium heat.Transfer the final concentrate to a dish and dry it using an electric drying oven until only the solute remains as a powder. Now, weigh the obtained powder, and dissolve it in sterile distilled water to create the daily dosage for mice. To prepare five amino salicylic acid, weigh 64 milligrams of five amino salicylic acid powder, and dissolve it in 200 milliliters of sterile distilled water.

六君子汤（LJZD）治疗溃疡性结肠炎-结直肠癌（UC-CRC）的制备

Establishment of an Ulcerative Coloproctitis-Colorectal Cancer (UC-CRC) Model to Evaluate the Therapeutic Effect of Chinese Medicine

To begin, gently position the mouse with its belly up and the head slightly down. Grasp the back skin to secure the abdominal area and insert a one milliliter syringe needle, one centimeter to the right of the midline of the abdomen at the intersection of the line, connecting both thighs. Gradually insert the one milliliter syringe needle to a depth of three to five millimeters under the skin.Insert the needle approximately 0.3 to 0.5 millimeters into the abdominal cavity at a 45 degree angle. Once the needle tip passes through the abdominal muscle, notice a sudden decrease in resistance. Slowly inject 0.1 milliliters of the azoxymethane injection solution per 10 grams of body weight into the mice.Subsequently, retract the syringe gently. Then add 500 milliliters of sterile distilled water to 10 grams of DSS. To prepare a 2%DSS solution.Mix the solution with a vortex mixer and store at four degrees Celsius until use, administer the prepared LJZD and five amino salicylic acid solution daily at a consistent time via gastric gavage to the respective groups at week seven and week 15.

建立溃疡性结肠直肠炎-结直肠癌（UC-CRC）模型评估中药的治疗效果

Evaluation of an Ulcerative Coloproctitis-Colorectal Cancer (UC-CRC) Model and Efficacy of Liujunzi Decoction (LJZD)

To begin, establish the UC-CRC mouse model and administer the prepared LJZD and 5-ASA solution to the respective groups. After the drug treatment, monitor the weight of mice daily from the beginning of adaptive feeding until the end of drug treatment. Diligently observe and document the fecal consistency of each experimental mouse, categorizing it as normal, loose stools or watery diarrhea.Carefully document any fecal bleeding observed in the experimental animals, categorizing it into no bleeding, minimal bleeding, or visible blood in the stool. To separate the colorectal tissue, maintain the euthanized mice within a cryogenic anatomical environment. Secure the mice in a supine position.Using scissors, trim the lower abdominal hair and sterilize the area with ethanol. Using eyelid forceps, gently grasp the intersection point between the two thigh roots and the abdominal midline. Then use scissors to create a one to 1.5 centimeters long transverse incision.Perform a longitudinal incision along the median line of the abdomen, starting from the midpoint of the transverse incision to the xiphoid process. Carefully detach the peri-colorectal tissues in the direction towards the anus, separating the colorectum rom the surrounding tissue. Gently push aside the abdominal skin to fully expose the colorectum.Using eyelid forceps, extract the colorectum from the abdominal cavity and section it from the anus to the cecum. Preserve the collected colorectal tissues in saline at four degrees Celsius. The UC-CRC model group had a significant weight loss compared to the control group which was alleviated by LJZD treatment.LJZD treatment improved the disease activity index compared with the UC-CRC model group.

作者聚焦： 六君子汤作为结肠直肠炎癌症的中医治疗方法

作者聚焦：六君子汤作为结肠直肠炎癌症的中医治疗方法