Takeda Masahiro, Miyahara Katsumi, Akazawa Chihiro, Lane Geoffrey J, Yamataka Atsuyuki
Department of Pediatric General and Urogenital Surgery, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan.
Department of Biochemistry and Biophysics, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Pediatr Surg Int. 2017 Aug;33(8):883-886. doi: 10.1007/s00383-017-4112-5. Epub 2017 Jun 10.
We used non-Hirschsprung's disease (HD) Sox10-Venus Transgenic mice (non-HDSV-mice), an endothelin receptor-B knockout mouse model of HD (HD-mice), and C57B6C3 wild controls (C-mice) to identify the correlation between the anorectal line (ARL) and successful transanal pull-through (TAPT).
In non-HDSV-mice, intestinal neural crest-derived cells can be visualized with Venus,-a green fluorescent protein-without histochemical staining. We exposed the anal canal in each non-HDSV-mouse and marked the ARL directly with red ink. Specimens of anus and rectum from HD- and C-mice were immunostained with sensory nerve markers substance P and calcitonin gene related peptide (CGRP) and Hematoxylin and Eosin.
Stereoscopic microscopy confirmed a squamous-columnar epithelial junction corresponding to the red ink in non-HDSV-mice. Fluorescence microscopy showed intense Venus expression proximal to the ARL and little enteric nerve expression distally. Substance P and CGRP expression were strong in the basal layer of the anal transitional zone (ATZ) in both HD- and C-mice; i.e., distal sensory innervation was normal in HD-mice.
The ARL delineated a distinct demarcation in sensory innervation that is normal even in HD-mice. Thus, the initial incision during TAPT should be based on the ARL because it is readily identifiable and intimately involved with bowel function.
我们使用非先天性巨结肠病（HD）的Sox10-维纳斯转基因小鼠（非HD-SV小鼠）、HD的内皮素受体B基因敲除小鼠模型（HD小鼠）以及C57B6C3野生对照小鼠（C小鼠）来确定肛门直肠线（ARL）与经肛门拖出术（TAPT）成功之间的相关性。
在非HD-SV小鼠中，肠道神经嵴衍生细胞可通过绿色荧光蛋白维纳斯进行可视化，无需组织化学染色。我们暴露每只非HD-SV小鼠的肛管，并用红色墨水直接标记ARL。对HD小鼠和C小鼠的肛门和直肠标本进行感觉神经标志物P物质和降钙素基因相关肽（CGRP）免疫染色以及苏木精和伊红染色。
立体显微镜证实非HD-SV小鼠中与红色墨水相对应的鳞状-柱状上皮交界处。荧光显微镜显示ARL近端维纳斯表达强烈，远端肠神经表达较少。HD小鼠和C小鼠肛门过渡区（ATZ）基底层P物质和CGRP表达均较强；即HD小鼠远端感觉神经支配正常。
ARL划定了感觉神经支配的明显界限，即使在HD小鼠中也是正常的。因此，TAPT期间的初始切口应基于ARL，因为它易于识别且与肠道功能密切相关。
