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Objective: To optimize the method for embedding multiple undecalcified mouse tibias in plastic blocks, improve the efficiency and stability of plastic embedding and reduce the detachment rate of plastic slides., Methods: Thirty undecalcified tibias from 15 B6 mice were used for plastic embedding after calcein labeling, fixation, dehydration and infiltration. The tibias were embedded in cylindrical plastic blocks with a diameter of 4 mm. For each bone, the 1/4 proximal tibia was cut off, and the remaining 3/4 was used for re-embedding. Five bones were embedded in a single block with each bone standing closely on the surface of a flat plate. The samples were randomized into control and experimental groups in all the processes of embedding, sectioning and staining. In the 3 groups with modified embedment, flowing CO 2 was added into the embedding solution, embedding solution was applied to the section surface, and the slides were heated at 95 ℃ for 15 min. The polymerization time, slide detachment rate, bone formation and osteoblast parameters were analyzed., Results: We prepared 6 plastic blocks, each containing 5 tibias, whose cross sections were on the same plane. The blocks were completely polymerized and suitable for sectioning. Flowing CO 2 into the embedding solution reduced the polymerization time and increased the rate of complete polymerization. Application of the embedding solution on the section surface significantly reduced the detachment rate of the sections ( P < 0.05) without affecting bone formation analysis ( P > 0.05). Heating the slides significantly lowered the detachment rate of the sections ( P < 0.05) without affecting osteoblast analysis ( P > 0.05)., Conclusions: The optimized method allows effective embedding of multiple undecalcified mice tibias in the same block and can be an ideal method for histological analysis of undecalcified bones.

Objective: To summarize the complications after endorectal pull-through radical operation, Soave procedure, for Hirschsprung disease., Methods: Clinical data of 286 cases with Hirschsprung disease who received Soave procedure and were proved by postoperative pathology in our hospital from February 2003 to February 2010 were analyzed retrospectively. All the patients were diagnosed with barium enema and anorectal manometry. Among them, 233 cases(81.5%) were identified during neonatal period. All the patients underwent cleansing enema and anus dilation immediately after definite diagnosis. Radical operation with Soave endorectal pull-through procedure, including simple transanal endorectal pull-through in 251 cases(87.8%), transabdominal approach in 17 cases (5.9%), laparoscopy-assisted endorectal pull-through in 18 cases (6.3%). The operations were performed in 54 cases (18.9%) within 3 months of age, in 183 cases (64.1%) between 3 and 6 months, in 38 cases(13.3%) between 6 months and one year, in 10 cases (3.5%) older than 1 year. The rectosigmoid was resected in 259 cases. Subtotal colectomy was performed in 25 cases, and total colectomy in 2 cases. Postoperative regular anal dilation lasted for 6 months. A total of 286 cases (male:250, female:36) were followed up for 2 to 5 years., Results: There was wound infection in 1 case, ileus in 1 cases, anastomotic stricture in 1 case, which was cured by continuous anal dilation. Two cases had constipation and received re-operation because of refractory to conservative therapy for 6 months. Perianal erosion was found in 63 cases(22.0%) and was healed within 3 months, except 2 children undergoing total colectomy. During follow-up, enterocolitis occurred in 11 cases(3.8%), including healing in 8 cases with conservative therapy, death in 1 case, and recurrent attacks in 2 cases. Soiling occurred in 45 cases(15.7%), among them, 5 cases presented in kindergarten and primary school. Morbidities of perianal erosion, enterocolitis and soiling were higher in infants undergoing operation within 3 months as compared to those more than 3 months [90.7%(49/63) vs. 6.0%(14/63), P=0.000; 9.3%(5/54) vs. 2.6%(6/232), P=0.022; 25.9%(14/54) vs. 13.4%(31/232), P=0.022]. Morbidity of perianal erosion enterocolitis was higher in infants undergoing subtotal or total colectomy as compared to those partial colon resection[51.9%(14/27) vs. 18.9%(49/259), P=0.000; 18.5%(5/27) vs. 2.3%(6/259), P=0.000]., Conclusions: Primary transanal endorental pull-through procedure can be performed in most children with Hirschsprung disease. Postoperative short-term complications are mainly perianal erosion and long-term ones are enterocolitis and soiling. Early diagnosis, colon irrigation, avoiding premature operation and anal dilation can decrease the morbidities of enterocolitis and soiling.