这项研究得到了穆尔西亚大学动物实验科和穆尔西亚大学妇产科科的支持。我们感谢所有的技术人员在 CEIB (中心实验 en 门多萨市 Biomédicas), 在穆尔西亚大学的动物实验部分, 谁在这个项目上合作。

这项研究是严格按照《关于国家卫生研究院的实验室动物的护理和使用指南》中的建议进行的, 并按照到达 (动物研究: 报告活体实验) 准则进行的。该协议获得了动物实验许可证, 符合指令 2010/63/欧盟的授权编号 A13170404 (Anexo 1)。所有的实验都是在欧洲联盟认证的实验室中进行的, 遵循国家动物伦理准则 (RD 53/2013, 欧盟指令 63/2010)。该议定书得到了穆尔西亚大学动物实验伦理学委员会的批准。
1. 动物制剂
注: 所有实验均得到穆尔西亚大学动物实验科和穆尔西亚大学妇产科学系的支持。
<ol><li>使用8周老的大鼠 (28 只雌性大鼠) 在所有实验中重200-250 克。 注意: 在这里, 我们还使用了5只怀孕的老鼠。</li>
	<li>为获得孕鼠, 笼八周老雌性和可育雄性大鼠和交配 17:00-23:00 h. 第二天早上阴道堵塞的识别被解释为交配成功.考虑1天的妊娠, 第二天后, 他们交配。</li>
	<li>在怀孕9、15和18天进行实验。</li>
	<li>两组的大鼠, 可以免费获得食物和水, 并保持正常12小时的光照/暗循环。</li>
	<li>在适应到设施条件至少两周后, 使用每天早晨的阴道细胞学来评估发情频率和规律性。 注: 二十八只大鼠与正常4至5天发情阶段选择纳入该研究。</li>
</ol>2. 大鼠成像的制备
注意: 图像是通过高分辨率经腹超声检查得到的。经阴道涂片细胞学检查, 可确定动情周期阶段。
<ol><li>在成像研究之前, 麻醉了2-3% 异氟醚气体在感应室的大坝。</li>
	<li>在超声检查中, 取出动物并立即将鼻口置于与麻醉系统连接的鼻子锥内, 并在 1.5-2% 异氟醚上维持动物。</li>
	<li>将皮草从肋缘移到尾腹, 用快船队和脱毛膏。</li>
	<li>将麻醉大鼠放置在一张加热桌上的仰卧位上, 以确保大鼠的安全, 并在影像疗程期间保证生理参数的最佳舒适和保持。所有的生理参数应与图像和实时采集的数据相结合, 通过超声应用。</li>
	<li>轻轻插入直肠探头 (在润滑后) 以监视体温 (37.5 ° c ±0.5 ° c)。</li>
	<li>将传感器 (30 Hz) 放在固定架上, 并使用手动操纵杆或手将其沿垂直轴和水平轴 (向前-向后和侧面) 移动。</li>
	<li>用石油为基础的眼霜, 以避免干燥过程中的老鼠的眼睛。</li>
</ol>3. 考试程序
注意: rats´生殖器官的解剖: 阴道位于膀胱的背部, 分为两个子宫角, 延伸至肾脏。卵巢通过输卵管连接到子宫角 (图 1)。子宫位于肾脏后部的区域。
<ol><li>
子宫的鉴别
	<ol>
<li>通过用膀胱作为地标来找到宫颈, 跟随宫颈找到左、右子宫角的分支。</li>
		<li>通过选择 "B 模式" 切换到二维查看/视频。用矢状中线图像测量中峡区各子宫角的前后径。使用超声系统软件检测测量。</li>
		<li>在 "B 型" 的矢状中线图像上, 测量子宫内膜腔的子宫内膜厚度, 从回声边界到回声边界。</li>
		<li>通过选择 "彩色多普勒", 改变彩色多普勒模式。使用彩色多普勒识别子宫内膜的血供, 测量子宫内膜的血流量。在彩色多普勒模式下选择以下参数: 高通滤波器设置在4赫兹, 脉冲重复频率设置在4和48赫之间, 脉冲多普勒门设置在0.2 到 0.5 mm 之间。</li>
	</ol>
</li>
	<li>
子宫动脉血流的鉴别
	<ol>
<li>获得多普勒波形的子宫动脉附近的侧下缘的宫内-颈部交界靠近髂动脉两侧。</li>
		<li>在多普勒模式下使用以下参数: 高通滤波器设置在6赫兹, 脉冲重复频率设置在4和48赫之间, 脉冲多普勒门设置在0.2 到 0.5 mm 之间。</li>
		<li>注意对齐血流和多普勒光束以最小化多普勒角。记录多普勒光束和血管之间的夹角。超出60°角度的值是不准确的, 应避免6。</li>
		<li>从三连续周期测量峰值收缩速度和舒张末期速度 (舒张)。然后计算每个子宫角的收缩到舒张期 (舒张) 和阻力指数 (RI) (舒张)/埃因霍温) 值。</li>
		<li>测量5例妊娠大鼠子宫动脉血流量 9th、15th、18th日。</li>
	</ol>
</li>
	<li>
卵巢动脉血流的鉴定 注: 雌性大鼠卵巢位于动物两侧的肾脏侧面, 并居住在子宫角末端的脂肪垫 (图 1)。<ol>
<li>要想象卵巢, 从探针开始在一个横向的平面, 并把它放在动物的侧面, 略低于肋骨。肾脏和脂肪垫有一个回声的外观相比, 卵巢。</li>
		<li>测量卵巢和卵泡的外边界。每个图像的刻度上的数字以毫米为单位, 增量为 0.1 mm。 注: 彩色多普勒模式和功率多普勒模式成像有助于识别卵巢强度和定向流。</li>
	</ol>
</li>
</ol>4. 研究的设计
<ol><li>用阴道涂片细胞学检查发情周期。</li>
	<li>把所有老鼠分成两组。对于组1或前受精 (或 periovulatory), 包括在 proestrus 和发情周期阶段的大鼠。对于2组或后育, 包括早期发情和晚期发情周期阶段的大鼠。</li>
	<li>监测和比较1和2组中峡区各子宫角的前后径。</li>
	<li>监测和比较1和2组子宫内膜的厚度和特征。</li>
	<li>监测和比较的大小 (最大直径) 和特点的卵巢, 并本地化任何 periovulatory 卵泡的两个卵巢组1和2。</li>
	<li>监测和比较1和2组的子宫动脉血流。</li>
	<li>监测和比较怀孕大鼠的子宫动脉血流在不同孕期 (9、15和18的妊娠期)。</li>
	<li>使用 SPSS 执行统计分析. 目前的数据为平均±标准差 (SD) 或中值与分范围。使用不同组之间的学生 t 检验分析结果。小于0.05 的 P 值被认为是统计学上的显著差异。</li>
</ol>

<ol>
	<li>Hunter, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose-Derived+Stromal+Vascular+Fraction+Cell+Effects+on+a+Rodent+Model+of+Thin+Endometrium.">Adipose-Derived Stromal Vascular Fraction Cell Effects on a Rodent Model of Thin Endometrium.</a> PLoS ONE. 10 (12), e0144823 (2015).</li><li>Wang, H., Dey, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roadmap+to+embryo+implantation:+clues+from+mouse+models.">Roadmap to embryo implantation: clues from mouse models.</a> Nat Rev Genet. 7 (3), 185-199 (2006).</li><li>Pistner, A., Belmonte, S., Coulthard, T., Blaxall, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+Echocardiography+and+Ultrasound+Imaging.">Murine Echocardiography and Ultrasound Imaging.</a> J Vis Exp. (42), (2010).</li><li>Lohmiller, J. J., Swing, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproduction+and+Breeding.">Reproduction and Breeding.</a> The Laboratory Rat. , 147-164 (2006).</li><li>Jaiswal, R. S., Singh, J., Adams, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+ultrasound+biomicroscopy+for+monitoring+ovarian+structures+in+mice.">High-resolution ultrasound biomicroscopy for monitoring ovarian structures in mice.</a> Reprod Biol Endocrinol. 7 (69), (2009).</li><li>Kim, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+Fetal+Echocardiography.">Murine Fetal Echocardiography.</a> J Vis Exp. (72), (2013).</li><li>Jing, Z., Qiong, Z., Yonggang, W., Yanping, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+bone+marrow+mesenchymal+stem+cells+improve+regeneration+of+thin+endometrium+in+rat.">Rat bone marrow mesenchymal stem cells improve regeneration of thin endometrium in rat.</a> Fertil Steril. 101 (2), 587-594 (2014).</li><li>Mu, J., Adamson, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+in+hemodynamics+of+uterine+artery,+utero-+and+umbilicoplacental,+and+vitelline+circulations+in+mouse+throughout+gestation.">Developmental changes in hemodynamics of uterine artery, utero- and umbilicoplacental, and vitelline circulations in mouse throughout gestation.</a> Am J Physiol Heart Circ Physiol. 291 (3), H1421-H1428 (2006).</li><li>Anderson, C. M., Lopez, F., Zhang, H. Y., Pavlish, K., Benoit, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+uteroplacental+perfusion+alters+uterine+arcuate+artery+function+in+the+pregnant+Sprague-Dawley+rat.">Reduced uteroplacental perfusion alters uterine arcuate artery function in the pregnant Sprague-Dawley rat.</a> Biol Reprod. 72 (3), 762-766 (2005).</li><li>Hongmei, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+Molecular+Imaging+of+Vascular+Endothelial+Growth+Factor+Receptor+2+Expression+for+Endometrial+Receptivity+Evaluation.">Ultrasound Molecular Imaging of Vascular Endothelial Growth Factor Receptor 2 Expression for Endometrial Receptivity Evaluation.</a> Theranostics. 5 (2), 206-217 (2015).</li></ol>

作者没有什么可透露的。

由于这项研究所需的程序性修改和故障排除, 尽管我们的目的是确定的所有阶段的发情周期的大鼠使用超声波, 我们无法找到任何显著的差异。我们推测, 这些困难可能是因为发情周期只持续几天的老鼠, 不同于妇女的周期。我们确信所有的测量都是在正确的时间进行, 以确定任何差异。因此, 我们将发情周期阶段重新组合为两个阶段, 以评估任何差异, 但没有观察到。正如所料, 我们发现每个子宫角?...

大鼠动物模型已广泛应用于生殖医学, 包括胚胎和卵巢移植的1,2。然而, 在过去, 对啮齿动物生殖系统发育的研究是基于对切除组织的一次性组织学检查, 而在大鼠3中, 对日间生殖器官变化的纵向研究是不可能的。超声在人类辅助生殖技术中的应用已有30年的历史, 但这一宝贵的技术在最近才被广泛应用于大鼠。
我们的目标是建立一个评价大大鼠生殖器官的超声方法, 设计生殖和妇科调查的实验模型, 并证明这一程序, 因为据我们所知,目前没有关于此过程的可视化出版物。我们描述了对雌性大鼠生殖系统进行超声检查的程序, 以及用高清晰度超声对解剖和子宫动脉血流的超声表现。我们监测不同发情周期的非妊娠期动物子宫内膜、卵巢和子宫动脉血流的特点, 以评价其在子宫膜厚度、卵巢形态和子宫血流中的显著差异。不同阶段的发情周期, 类似于妇女。我们使用了 70 MHz 频率和分辨率级别为30µm 的高品质超声设备.我们的另一个目的是评估怀孕大鼠子宫血流阻力的变化。这种技术允许研究生殖器官的日常变化而不牺牲动物。
超声对大鼠的应用有若干技术难点。这些困难包括: 大鼠子宫内膜比人类女性更薄4。大鼠卵巢成像困难的原因是大鼠的皮肤和腹壁肌组织较厚, 导致超声5近乎完全衰减, 而子宫动脉在非妊娠期更难发现。鼠.我们已经解决了程序中的许多技术问题, 对于那些仍然存在的问题, 我们将展示如何将它们最小化。
在不需要牺牲动物的情况下, 成功地监测大鼠生殖器官的超声变化, 将为将来建立生殖医学和其他外科手术的动物模型开辟可能性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Vevo3100 high-resolution in vivo micro-imaging system*</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>Vevo integrated rail system including physiological monitoring unit.</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>MX400 Transducter</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>Vevo Lab Software</td>
			<td>Visual Sonics, inc.</td>
			<td></td>
			<td>www.visualsonics.com/products</td>
		</tr>
		<tr>
			<td>HSD: Sprague Dawley SD</td>
			<td>Envigo, inc.</td>
			<td></td>
			<td>Rat strain</td>
		</tr>
		<tr>
			<td>Lubricating Gel</td>
			<td>General Supply</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CIBERTEC CA-EAC20 Anesthesia Trolley System</td>
			<td>Cibertec S.A&#160;</td>
			<td></td>
			<td>Anesthesia Machine</td>
		</tr>
		<tr>
			<td>Ecogel 100 ultrasound gel</td>
			<td>Eco-Med Pharmaceuticals Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removal lotion (Nair)&#160;</td>
			<td>General Supply</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Esteve Veterinaria</td>
			<td></td>
			<td>Inhalatory anesthesia</td>
		</tr>
		<tr>
			<td>* Required software is Vevo software including B-Mode application, pulse wave Doppler application, and vascular strain analysis tools package.</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

ultrasonography in experimental reproductive investigations on rats

随着辅助生殖技术的发展和人类研究的伦理局限性, 大鼠动物模型在生殖医学中得到了广泛的应用。过去, 对啮齿动物生殖系统发育的研究一直是基于对切除组织的一次性组织学检查。近年来, 随着高分辨率经腹超声的发展, 可以对大鼠的生殖器官进行高质量的超声检测, 从而为研究生殖系统提供了一种新的方法。利用高分辨率的超声系统获得图像。对28八周大的非孕鼠和5例妊娠大大鼠进行妇科超声检查。我们描述了在发情周期的不同阶段, 如何在典型的视图中识别生殖系统和相关结构的器官。彩色血流多普勒用于测量子宫动脉血流, 评价不同孕期子宫血流模式的变化。我们已经证明, 超声探测是一种有用的方法来评价变化的内部生殖器官。它的使用提高了进行额外实验的可能性, 包括医疗或外科手术, 并提供了监测超声改变内脏器官而不牺牲动物的能力。

前后子宫角直径或子宫角的两侧子宫内膜厚度 (表 1) 无显著差异。与2组比较, 1 组的平均子宫内膜厚度较厚, 但在两组 (图 3) 之间未发现显著差异 (P > 0.05)。然而, 我们发现子宫内的液体 (在8出28大鼠) 附近的发情周期的变化相关的内膜形态学 (图 2)。
多普勒研究还显示, 非妊娠大鼠子宫角两侧或不同发情周期的血流速度?...

Watch this Scientific Journal Video about Ultrasonography in Experimental Reproductive Investigations on Rats at JoVE.com

Ultrasonography in Experimental Reproductive Investigations on Rats

该手稿描述了超声对雌性大鼠进行生殖和妇科研究的实验模型的效用。一步一步的解释如何进行超声评价显示。

大鼠实验性生殖研究中的超声检查

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in ...

ultrasonography-in-experimental-reproductive-investigations-on-rats

Research

JoVE Journal

Medicine

14.2K 次观看.  The First Affiliated Hospital of Anhui Medical University. 该手稿描述了超声对雌性大鼠进行生殖和妇科研究的实验模型的效用。一步一步的解释如何进行超声评价显示。

视频: Ultrasonography in Experimental Reproductive Investigations on Rats

Acute Myocardial Infarction in Rats

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular diseases. Animal models that mimic human cardiac disease, such as myocardial infarction (MI) and ischemia-reperfusion (IR) that induces heart failure as well as pressure-overload (transverse aortic constriction) that induces cardiac hypertrophy and heart failure (Goldman and Tarnavski), are useful models to study cardiovascular disease. In particular, myocardial ischemia (MI) is a leading cause for cardiovascular morbidity and mortality despite controlling certain risk factors such as arteriosclerosis and treatments via surgical intervention (Thygesen). Furthermore, an acute loss of the myocardium following myocardial ischemia (MI) results in increased loading conditions that induces ventricular remodeling of the infarcted border zone and the remote non-infarcted myocardium. Myocyte apoptosis, necrosis and the resultant increased hemodynamic load activate multiple biochemical intracellular signaling that initiates LV dilatation, hypertrophy, ventricular shape distortion, and collagen scar formation. This pathological remodeling and failure to normalize the increased wall stresses results in progressive dilatation, recruitment of the border zone myocardium into the scar, and eventually deterioration in myocardial contractile function (i.e. heart failure). The progression of LV dysfunction and heart failure in rats is similar to that observed in patients who sustain a large myocardial infarction, survive and subsequently develops heart failure (Goldman). The acute myocardial infarction (AMI) model in rats has been used to mimic human cardiovascular disease; specifically used to study cardiac signaling mechanisms associated with heart failure as well as to assess the contribution of therapeutic strategies for the treatment of heart failure. The method described in this report is the rat model of acute myocardial infarction (AMI). This model is also referred to as an acute ischemic cardiomyopathy or ischemia followed by reperfusion (IR); which is induced by an acute 30-minute period of ischemia by ligation of the left anterior descending artery (LAD) followed by reperfusion of the tissue by releasing the LAD ligation (Vasilyev and McConnell). This protocol will focus on assessment of the infarct size and the area-at-risk (AAR) by Evan's blue dye and triphenyl tetrazolium chloride (TTC) following 4-hours of reperfusion; additional comments toward the evaluation of cardiac function and remodeling by modifying the duration of reperfusion, is also presented. Overall, this AMI rat animal model is useful for studying the consequence of a myocardial infarction on cardiac pathophysiological and physiological function.

The rat model of acute myocardial infarction (AMI) is useful to study the consequence of a MI on cardiac pathophysiological and physiological function.

大鼠急性心肌梗死

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular ...

科研

医学

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Experimental Human Pneumococcal Carriage

Experimental human pneumococcal carriage (EHPC) is scientifically important because nasopharyngeal carriage of Streptococcus pneumoniae is both the major source of transmission and the prerequisite of invasive disease. A model of carriage will allow accurate determination of the immunological correlates of protection, the immunizing effect of carriage and the effect of host pressure on the pathogen in the nasopharyngeal niche. Further, methods of carriage detection useful in epidemiologic studies, including vaccine studies, can be compared.
Aim
We aim to develop an EHPC platform that is a safe and useful reproducible method that could be used to down-select candidate novel pneumococcal vaccines with prevention of carriage as a surrogate of vaccine induced immunity. It will work towards testing of candidate vaccines and descriptions of the mechanisms underlying EHPC and vaccine protection from carriage1. Current conjugate vaccines against pneumococcus protect children from invasive disease although new vaccines are urgently needed as the current vaccine does not confer optimal protection against non-bacteraemic pneumonia and there has been evidence of serotype replacement with non-vaccine serotypes2-4.
Method
We inoculate with S. pneumoniae suspended in 100 &mu;l of saline. Safety is a major factor in the development of the EHPC model and is achieved through intensive volunteer screening and monitoring. A safety committee consisting of clinicians and scientists that are independent from the study provides objective feedback on a weekly basis.
The bacterial inoculum is standardized and requires that no animal products are inoculated into volunteers (vegetable-based media and saline). The doses required for colonization (104-105) are much lower than those used in animal models (107)5. Detecting pneumococcal carriage is enhanced by a high volume (ideally &gt;10 ml) nasal wash that is relatively mucus free. This protocol will deal with the most important parts of the protocol in turn. These are (a) volunteer selection, (b) pneumococcal inoculum preparation, (c) inoculation, (d) follow-up and (e) carriage detection.
Results
Our current protocol has been safe in over 100 volunteers at a range of doses using two different bacterial serotypes6. A dose ranging study using S. pneumoniae 6B and 23F is currently being conducted to determine the optimal inoculation dose for 50% carriage. A predicted 50% rate of carriage will allow the EHPC model to have high sensitivity for vaccine efficacy with small study numbers.

Experimental human pneumococcal carriage offers a natural model of carriage and a potential model for use in vaccine development. This technique is valuable yet complex and involves clinical risk by introducing a pathogen into a human. We have developed a detailed protocol.

人体实验肺炎球菌运输

Experimental human pneumococcal carriage (EHPC) is scientifically important because nasopharyngeal carriage of Streptococcus pneumoniae is both the major ...

Analyzing Beneficial Effects of Nutritional Supplements on Intestinal Epithelial Barrier Functions During Experimental Colitis

Inflammatory bowel diseases (IBD), including Crohn&#39;s disease and ulcerative colitis, are chronic relapsing disorders of the intestines. They cause severe problems, such as abdominal cramping, bloody diarrhea, and weight loss, in affected individuals. Unfortunately, there is no cure yet, and treatments only aim to alleviate symptoms. Current treatments include anti-inflammatory and immunosuppressive drugs that may cause severe side effects. This warrants the search for alternative treatment options, such as nutritional supplements, that do not cause side effects. Before their application in clinical studies, such compounds must be rigorously tested for effectiveness and security in animal models. A reliable experimental model is the dextran sulfate sodium (DSS) colitis model in mice, which reproduces many of the clinical signs of ulcerative colitis in humans. We recently applied this model to test the beneficial effects of a nutritional supplement containing vitamins C and E, L-arginine, and &#969;3-polyunsaturated fatty acids (PUFA). We analyzed various disease parameters and found that this supplement was able to ameliorate edema formation, tissue damage, leukocyte infiltration, oxidative stress, and the production of pro-inflammatory cytokines, leading to an overall improvement in the disease activity index. In this article, we explain in detail the correct application of nutritional supplements using the DSS colitis model in C57Bl/6 mice, as well as how disease parameters such as histology, oxidative stress, and inflammation are assessed. Analyzing the beneficial effects of different diet supplements may then eventually open new avenues for the development of alternative treatment strategies that alleviate IBD symptoms and/or that prolong the phases of remission without causing severe side effects.

Current treatments of inflammatory bowel diseases (IBD) aim at alleviating disease symptoms but may cause severe side effects. Thus, alternative strategies are being investigated in animal colitis models. Here, we explain how the beneficial effects of diet supplements on clinical IBD signs are analyzed in such a colitis model.

分析营养补充剂的有益作用于肠上皮屏障功能在实验性结肠炎

Inflammatory bowel diseases (IBD), including Crohn&#39;s disease and ulcerative colitis, are chronic relapsing disorders of the intestines. They cause ...

Effects of Allogeneic Platelet-Rich Plasma (PRP) on the Healing Process of Sectioned Achilles Tendons of Rats: A Methodological Description

This article describes the experimental procedures used to observe if PRP can positively affect tendon healing. There are 4 main steps to follow: induce a lesion in the Achilles tendon; prepare PRP and inject it (or the saline solution); remove the tendon; and perform biomechanical, molecular, and histological evaluations. At each step, all the procedures and methods are described in detail, so they can be reproduced easily.
Achilles tendons have been surgically sectioned (removal of a 5-mm long section). Afterwards, PRP or saline solution was injected to study whether PRP has a positive effect on the healing of the tendon. Three groups of 40 animals (a total of 120 rats were used in this study) were subdivided into 2 subgroups: PRP injection group and a saline injection control group. Rats were sacrificed at increasing time points (Group A: 5 days; Group B: 15 days; Group C: 30 days) and tendons were removed. 90 tendons underwent biomechanical testing before performing transcriptomic analysis and the 30 remaining tendons were submitted to histological analysis.

Effects of Allogeneic Platelet-Rich Plasma PRP on the Healing Process of Sectioned Achilles Tendons of Rats: A Methodological Description

This protocol describes the evaluation process of healing tendons in rats that have been injected with allogeneic platelet rich plasma (PRP) or saline solution after removing part of the Achilles tendon. The progress of tendon healing is evaluated at several time points using different types of analyses.

同种异体富血小板血浆 (PRP) 对大鼠分段腱愈合过程的影响: 方法学描述

This article describes the experimental procedures used to observe if PRP can positively affect tendon healing. There are 4 main steps to follow: induce a ...

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent hemorrhages at a later stage. The exact effects of plasma coagulation on liver tissue are only poorly examined. In our porcine model, the coagulation effects can be examined close to the clinical application. A combined laser Doppler flowmeter and spectrophotometer documents microcirculation changes during coagulation at 8 mm tissue depth noninvasively, providing quantifiable information about hemostasis beyond the subjective clinical impression. The temperature at coagulation site is assessed with an infrared thermometer prior and post coagulation and with a thermographic camera during coagulation, a measurement of the gas beam temperature is not possible due to the upper threshold of the devices. The depth of coagulation is measured microscopically on hematoxylin/eosin stained sections after calibration with an object micrometer and gives an exact information about the power setting-coagulation depth-relation. The sealing effect is examined on the bile ducts as it is not possible for a plasma coagulator to seal larger blood vessels. Burst pressure experiments are carried out on explanted organs to rule out blood pressure related effects.

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Here we present a protocol to experimentally assess plasma coagulation in liver tissue in vivo. In a porcine model, microcirculation is examined by laser Doppler, coagulation depth is measured histologically, temperature at coagulation site by infrared thermometer and thermographic camera, and duct sealing effect is documented by burst pressure experiments.

大动物模型肝组织血浆凝固的评价

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

大鼠胃旁路 (oagb) 的一次吻合

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

On-Site Sampling and Extraction of Brain Tumors for Metabolomics and Lipidomics Analysis

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still in need. In recent years, mass spectrometry has become a method of choice for untargeted profiling of discriminatory compound as potential biomarkers of a disease. Biofluids are generally considered as preferable matrices given their accessibility and easier sample processing while direct tissue profiling provides more selective information about a given cancer. Preparation of tissues for the analysis via traditional methods is much more complex and time-consuming, and, therefore, not suitable for fast on-site analysis. The current work presents a protocol combining sample preparation and extraction of small molecules on-site, immediately after tumor resection. The sampling device, which is of the size of an acupuncture needle, can be inserted directly into the tissue and then transported to the nearby laboratory for instrumental analysis. The results of metabolomics and lipidomics analyses demonstrate the capability of the approach for the establishment of phenotypes of tumors related to the histological origin of the tumor, malignancy, and genetic mutations, as well as for the selection of discriminating compounds or potential biomarkers. The non-destructive nature of the technique permits subsequent performance of routinely used tests e.g., histological tests, on the same samples used for SPME analysis, thus enabling attainment of more comprehensive information to support personalized diagnostics.

The manuscript presents on-site sampling of human brain tumors with solid phase microextraction followed by their biochemical profiling towards the discovery of biomarkers.

代谢组学和利皮多米分析脑肿瘤的现场采样与提取

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still ...

Preparation of Decellularized Kidney Scaffolds in Rats

Tissue engineering is a cutting-edge discipline in biomedicine. Cell culture techniques can be applied for regeneration of functional tissues and organs to replace diseased or damaged organs. Scaffolds are needed to facilitate the generation of three-dimensional organs or tissues using differentiated stem cells in vivo. In this report, we describe a novel method for developing vascularized scaffolds using decellularized rat kidneys. Eight-week-old Sprague-Dawley rats were used in this study, and heparin was injected into the heart to facilitate flow into the renal vessels, allowing heparin to perfuse into the renal vessels. The abdominal cavity was opened, and the left kidney was collected. The collected kidneys were perfused for 9 h using detergents, such as Triton X-100 and sodium dodecyl sulfate, to decellularize the tissue. Decellularized kidney scaffolds were then gently washed with 1% penicillin/streptomycin and heparin to remove cellular debris and chemical residues. Transplantation of stem cells with the decellularized vascular scaffolds is expected to facilitate the generation of new organs. Thus, the vascularized scaffolds may provide a foundation for tissue engineering of organ grafts in the future.

This protocol introduces a method to develop a scaffold using decellularized rat kidneys. The protocol includes decellularization and recellularization processes to confirm bioavailability. Decellularization is performed using Triton X-100 and sodium dodecyl sulfate.

在大鼠中准备去细胞化肾脏脚手架

Tissue engineering is a cutting-edge discipline in biomedicine. Cell culture techniques can be applied for regeneration of functional tissues and organs ...

A Swin Transformer-Based Model for Thyroid Nodule Detection in Ultrasound Images

In recent years, the incidence of thyroid cancer has been increasing. Thyroid nodule detection is critical for both the detection and treatment of thyroid cancer. Convolutional neural networks (CNNs) have achieved good results in thyroid ultrasound image analysis tasks. However, due to the limited valid receptive field of convolutional layers, CNNs fail to capture long-range contextual dependencies, which are important for identifying thyroid nodules in ultrasound images. Transformer networks are effective in capturing long-range contextual information. Inspired by this, we propose a novel thyroid nodule detection method that combines the Swin Transformer backbone and Faster R-CNN. Specifically, an ultrasound image is first projected into a 1D sequence of embeddings, which are then fed into a hierarchical Swin Transformer. 
The Swin Transformer backbone extracts features at five different scales by utilizing shifted windows for the computation of self-attention. Subsequently, a feature pyramid network (FPN) is used to fuse the features from different scales. Finally, a detection head is used to predict bounding boxes and the corresponding confidence scores. Data collected from 2,680 patients were used to conduct the experiments, and the results showed that this method achieved the best mAP score of 44.8%, outperforming CNN-based baselines. In addition, we gained better sensitivity (90.5%) than the competitors. This indicates that context modeling in this model is effective for thyroid nodule detection.

Here, a new model for thyroid nodule detection in ultrasound images is proposed, which uses Swin Transformer as the backbone to perform long-range context modeling. Experiments prove that it performs well in terms of sensitivity and accuracy.

基于Swin变压器的超声图像甲状腺结节检测模型

In recent years, the incidence of thyroid cancer has been increasing. Thyroid nodule detection is critical for both the detection and treatment of thyroid ...