ObjectiveTo investigate the effectiveness of posterior sagittal anorectoplasty (PSARP) and total urogenital mobilization in treatment of cloacal malformation. MethodsThree girls with cloacal malformation were admitted between June 2010 and June 2013. The age was 2 years, 5 months, and 4 months, respectively. All patients received previous separated sigmoid colostomy. PSARP and total urogenital mobilization were performed to correct the malformation. Colostomy stoma was closed after confirming the patent of rectum and anus. Dilation of anus was sustained for 6 months after anorectoplasty. ResultsOperations were successfully completed in 3 cases. The durations of operation were 3 hours and 40 minutes, 4 hours and 20 minutes, and 3 hours and 50 minutes; the blood loss was 10, 20, and 10 mL; and the time of colostomy stoma closure was 3, 5, and 3 months, respectively. Three cases were followed up 13, 18, and 4 months, respectively; no anal stenosis or rectal prolapse occurred. They had normal micturition; the B ultrasonic examination showed that the residual urine volume was less than 10 mL. At 12 months after anorectoplasty, the Wexner scores were 1 and 5 in 2 cases followed up 13 and 18 months; according to Krickenbeck postoperative effect, 2 cases had defecation and no constipation; 1 case had soiling (grade 1). ConclusionPSARP and total urogenital mobilization should be the first choice in treatment of cloacal malformation with common channel less than 3 cm.
ObjectiveTo explore the effectiveness of thoracoscopic surgery for treating late-presenting congenital diaphragmatic hernias and summarize the experience. MethodsBetween October 2012 and February 2015, 21 children with late-presenting congenital diaphragmatic hernias underwent thoracoscopic surgery. Of the 21 cases, 12 were girls and 9 were boys with a median age of 1 year and 3 months (range, 2 months to 8 years). Eight patients had obvious symptom in the initial stage:shortness of breath and dyspnea; 13 cases were found occasionally through chest radiography. Of 21 cases, 17 had left diaphragmatic hernias and 4 had right diaphragmatic hernias. The emergency surgery was performed in 5 cases because oppressed obviously and selective operation in 16 cases. Hernial sac existed in 5 cases; there were 19 cases of Bochdalek's hernia and 2 cases of Morgagni's hernia. The size of diaphragmatic defect ranged from 3 cm×2 cm to 5 cm×5 cm. ResultsThe operation time was 35-80 minutes (mean, 50 minutes), and intraoperative blood loss was 3-5 mL (mean, 3.8 mL). Primary healing of incision was obtained. Postoperative abdominal distension and pneumothorax occurred in 12 and 2 cases respectively. The follow-up time was 1-3 years (mean, 20 months). All the cases had a good recovery and satisfactory appearance of the thoracic incision. The symptoms and signs of shortness of breath and dyspnea disappeared. There was no recurrence and chest infection. ConclusionUnder the conditions of mastering operative indications strictly, thoracoscopic repair for late-presenting congenital diaphragmatic hernia is safe and feasible. It can facilitate the procedure and decrease the recurrence rate relying on intraoperative application of hernia repair needle, knot pusher-assistant, and reasonable processing defect periphery.
ObjectiveTo study the growth of adipose-derived stem cells (ADSCs) planted in three-dimensional (3D) materials, a 3D cultured ADSCs system based on microbial transglutaminase (mTG) enzyme crosslinked gelatin hydrogel was constructed. MethodsADSCs were isolated from the subcutaneous adipose tissue of a Sprague Dawley rat by collagenase digestion and centrifugation, and were cultured for passage. The mTG enzyme crosslinked gelatin hydrogel was firstly synthesized by mixing gelatin and mTG, and then the ADSCs were encapsulated in situ (2D environment) and cultured in the 3D materials (3D environment). The morphology and adhesion of cells were observed by inverted phase contrast microscope. In addition, HE staining and Masson staining were carried out to observe the distribution of cells in the material. Living and death situation of ADSCs in the materials was observed by fluorescence microscope and laser scanning confocal microscopy. Scanning electron microscopy was used to observe the adhesion of ADSCs on hydrogel surface. Alamar-Blue method was used to detect the proliferation of ADSCs in the hydrogel. Moreover, the results were compared between the cells cultured in 2D environment and those in 3D environment. ResultsThe result of 2D culture showed that ADSCs grew well on the hydrogel surface with normal functioning and had good adhesion. The results of 3D culture showed that ADSCs grew well in 3D cultured mTG enzyme crosslinked gelatin hydrogel, and presented 3D shape. Cells obviously extended in all directions. The number of apoptotic cells was very small. The cells of 3D culture at each time point was significantly less than that of the conventional culture cells, difference was statistically significant (P < 0.05). But after 8 days culture, the proliferation of the cells cultured in the mTG enzyme crosslinked gelatin hydrogel increased more quickly. ConclusionADSCs can grow well with good adhesion and show high viability in 3D culture system constructed by mTG enzyme crosslinked gelatin hydrogel.
Bone marrow-derived mesenchymal stem cells (BMSCs) for repairing damaged heart tissue are a new kind of important treatment options because of their potential to differentiate into cardiomyocytes. We in this experiment investigated the effect of different electrical stimulation time on the expression of myocardial specificity gene and protein in rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. The rBMSCs of second or third generation were randomly divided into three groups, i.e. electrical stimulation (ES) group, 5-Azacytidine (5-Aza) group and the control group. The rBMSCs in the ES groups with complete medium were exposed to 2 V, 2 Hz, 5 ms electrical stimulation for 0.5 h, 2 h, 4 h, and 6 h respectively every day for 10 days. Those in the 5-Aza group were induced by 5-Aza (10 μmol/L) for 24 h, and then cultured with complete medium for 10 days. Those in the control group were only cultured with complete medium, without any treatment, for 10 days. The rBMSCs' morphological feature in each group was observed with inverted phase microscope. The mRNA expression of myocyte-specific enhancer factor 2C (MEF-2C) and connexin 43 (Cx43) were examined with Real-Time quantitative PCR and the protein expression of MEF-2C, Cx43 were detected with Western Blot method. The results showed that the mRNA expression level of the MEF-2C, Cx43 and the protein expression level of MEF-2C, Cx43 were significantly higher in the ES group and 5-Aza group than those in the relative control group (P < 0.05). It suggests that electrical stimulation could play a part of role in the induction of the rBMSCs to differentiate into the cariomyocyte-like cells in vitro and the effectiveness of the electrical stimulation with 2 h/d had the best in our experiement. But the mechanism how electrical stimulation promotes the differentiation of rBMSC into cardiomyocyte is still unclear.
ObjectiveTo investigate how to shorten the learning curve of the laparoscopic pancreaticoduodenectomy (LPD). MethodsClinical data of 5 patients who underwent the LPD in our hospital from May 2015 to November 2015 were retrospectively analyzed. ResultsThe mean age of 58.8 years old. There were four patients who were diagnosed with periampullary tumor, one patient was distal bile duct carcinoma. The median operative time was 588 min, the average blood loss was 290 mL, the time of feeding was 5 days, the mean hospital stay was 25 days. One case died of cardiovascular event on postoperative day 1. One patient had postoperative bleeding after LPD, who recovered smoothly after reoperation for hemostasis laparoscopiclly. Conciusions LPD needs basic learning curve. The key of this procedure are appropriate treatment of pancreatic head and digestive tract reconstruction. Rich operative experience of surgeon in pancreaticoduodenectomy, optimization of the operation process, skilled in laparoscopic procedures, appropriate cases, appropriate perioperative management, and steady surgical team are also important factor for the success of LPD and shorten learning curve.