先天性巨结肠患儿切除肠管肠黏膜ENS干细胞的初步鉴定

目的 我们已经证实p75NTR阳性细胞在HD(Hirschsprung's disease)患儿切除肠管的移形段和扩张段肌间丛及黏膜下丛存在,本研究拟对肠黏膜层进行研究.另外,为了明确Metzger等在肠黏膜活检标本中分离体外培养出的肠黏膜ENS干细胞是胚胎残留还是其他肠黏膜干细胞在诱导分化过程中转化而来,本研究选用了不同时期肠神经干细胞标记物(早期:SOX10和Nestin中晚期:TUJ1;晚期:GFAP),对病理已证实的婴儿组(年龄≤1岁)和幼儿组(年龄＞1岁)HD患儿的切除肠管标本连续切片后进行研究.同时对黏膜层、黏膜下丛及肌间丛的SOX10、Nestin、TUJ1及GFAP强阳性表达率进行综合分析,明确GNCSCs在三层间的存在差异及其与年龄间的关系,以指导临床取材.方法 收集临床诊断及病理确诊的HD患儿手术切除标本15例,分为婴儿组(8例)及幼儿组(7例),长段型(2例)及常见型(13例).沿肠管纵轴各取材一处并将其分为3组:第一组为狭窄段肠管,第二组为移行段肠管,第三组为扩张段肠管.对照组选用切除扩张段肠管的近端肠管,阴性对照应用TBS缓冲液作为一抗.应用S-P法分析四个指标在各标本、各部位的表达情况.结果 SOX10、Nestin和TUJ1在移行段和扩张段黏膜层均有强阳性表达,且在黏膜固有层呈散在分布,但主要位于近黏膜肌层,GFAP在移行段和扩张段黏膜层未见阳性表达.肌间丛SOX10、Nestin、TUJ1和GFAP免疫阳性细胞的强阳性表达率高于黏膜下丛,黏膜下丛强阳性表达率高于黏膜层.婴儿组患儿在扩张段肌间丛SOX10和Nestin强阳性表达率较幼儿组高.婴儿组扩张段黏膜层SOX10和Nestin强阳性表达率较幼儿组高.结论 在HD患儿有神经节段肠管肠黏膜可能存在ENS干细胞或前体细胞,且在肠黏膜固有层呈散在分布,但主要定位于近黏膜肌层.SOX10、Nestin、TUJ1和GFAP在HD患儿肌间丛表达水平较黏膜下丛高,而黏膜下丛表达水平较黏膜层高.婴儿组扩张段黏膜层SOX10和Nestin表达水平可能较幼儿组高,随年龄增长可能逐渐降低.初步判定存在于肠黏膜的ENS干细胞或前体细胞可能来源于肠神经嵴细胞.

Abstract：

Objective To identify the enteric nervous system (ENS) stem cells in intestinal mucosa of the patients with Hirschsprung's disease (HD). Methods Fifteen patients were pathologically diagnosed with Hirschsprung's disease. They underwent surgery to remove the aganglionic intestines.The removed intestines were collected for this study. Of the 15 patients, 8 were infants (age≤1 year)and 7 were toddlers (age＞ 1year). Two were long-segment HD, and 13 were short-segment HD.Three pieces of tissues were collected from the stenotic segment, transitional zone and dilated segment of the removed intestines. The intestines proximate to the dilated segment were selected as controls.Immunohistochemical staining of stem cell markers including SOX10, Nestin, TuJ1 and GFAP was performed to indentify the ENS stem cells in intestinal mucosa. The TBS buffer was used as the negative control of the primary antibody. The S-P method was applied to analyze the expressions of the 4stem cell markers through the intestine. Results The expressions of SOX10, Nestin, and TUJ1 in the mucosa of the transitional zone and dilated segment were strong. And scattered SOX10, Nestin, and TUJ1 positive cells were observed in the lamina propria. But most of the SOX10, Nestin, and TUJ1 positive cells located in mucosa closed to the muscularis mucosa. The expression of GFAP in the mucosa of the transitional zone and dilated segment was negative. The expressions of SOX10, Nestin,TUJ1 and GFAP in myenteric plexus were stronger than those of submucosal plexus (SOX10 expression, infant 50% vs 31.3%, toddler 35. 7% vs 21.4%; Nestin expression, infant 43. 8% vs 37. 5%,toddler 28. 6% vs 14. 2%; TUJ1 expression, infant 37. 5% vs 18. 5%; toddler 28. 6% vs 28. 6%;GFAP expression 25.0 % vs 18. 5 % ; toddler 57. 1% vs 35. 7 % ; P ＜ 0. 05 ). The expressions of SOX10, Nestin and TUJ1 were stronger in submucosal plexus than those in mucosa (SOX10 expression, infant 31.3% vs 25. 0%, toddler 21.4% vs 7. 1% ; Nestin expression, infant 25. 0% vs 18. 8%,toddler 14. 3% vs 14. 3%; TUJ1 expression, infant 18. 5% vs 12. 5 %, toddler 28. 6% vs 7. 1%; P＜0. 05). The expressions of SOX10 and Nestin in myenteric plexus of the dilated segment in infants were stronger than thoset in toddler (SOX10, 75. 0% vs 42. 8% ; Nestin, 62. 5% vs 42. 8%, P＜0. 05). The expressions SOX10 and Nestin in mucosa of the the dilated segment in infants were stronger than those in toddler (SOX10, 37. 5% vs 14. 3%; Nestin, 25. 0% vs 14. 3%, P＜0. 05). Conclusions The ENS stem cells may exist in the mucosa of the ganglionic intestines of HD patients, whichare scattered and mostly located in the mucosa near muscularis mucosa. In HD patients, the expressions of SOX10, Nestin, TUJ1 and GFAP in myenteric plexus are higher than those in submucosal plexus. However, the expressions of SOX10, Nestin, TUJ1 and GFAP in the submucosal plexus are higher than those in the mucous layer. The expressions of SOX10 and Nestin in mucosa of the dilated segment in infants are higher than those in toddlers, and it decreases with age. These observations suggest the ENS stem cells or precursor cells in the gut mucosa may be derived from the gut neural crest cells.     

微等离子体射频技术在急诊面部美容缝合术后的应用

目的:探讨急诊面部外伤患者在美容缝合术后应用微等离子体射频治疗的效果。方法:选择2016年10月-2017年9月收治的40例面部外伤患者随机分为2组,每组20例。观察组患者于急诊美容缝合术后行微等离子射频治疗,对照组仅接受美容缝合治疗。结果:所有患者经美容缝合术后均达到一期愈合,经微等离子射频治疗后,面部瘢痕明显改善,观察组患者2例完全恢复,10例明显改善,8例中度改善,无效0例,无严重并发症发生。结论:急诊面部外伤患者在美容缝合术后应用微等离子体射频治疗可进一步改善面部外观,治疗效果较好,不良反应少,值得临床推广应用。     

胃复安加速腹腔镜阑尾切除术后胃肠功能恢复的临床观察

目的:观察胃复安对腹腔镜阑尾切术后胃肠道功能恢复的临床效果。方法:选取2021年1－7月四川省色达县人民医院行腹腔镜阑尾切除术的患者60例,随机分为对照组和观察组,每组各30例。对照组术后仅给予常规抗感染、抑酸、补液等治疗;观察组在上述治疗的基础上加用胃复安。以胃肠功能如首次闻及肠鸣音、肛门首次排气排便等作为观察指标,记录数据进行统计学比较,评价胃复安对腹腔镜阑尾切除术后胃肠功能恢复的临床疗效。结果:观察组患者术后首次闻及肠鸣音时间、首次排气时间、首次排便时间及住院时间明显短于对照组,差异有统计学意义(P<0.05);两组术后并发症总发生率比较,差异无统计学意义。但观察组术后出现恶心、呕吐、腹胀例数少于对照组。结论:胃复安可加速腹腔镜下阑尾切除术后患者胃肠功能的恢复。     

香青兰中总黄酮和单体的含量测定

目的:考察香青兰中单体(刺槐素-7-o-β-D-葡萄糖苷)、总黄酮的含量。方法:采用紫外分光光度法(UV)测定总黄酮含量;采用高效液相色谱法(HPLC)测定单体含量,RP-C18柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%磷酸水(26∶74),流速为1.1 mL·min-1,柱温室温,检测波长为326 nm。结果:单体(刺槐素-7-o-β-D-葡萄糖苷)含量为0.33%,回收率为98.62%;总黄酮含量为1.90%,回收率为97.59%。结论:所建方法准确、简便,为科学地评价香青兰药材的质量提供依据。     

颅脑损伤首次CT检查阴性的原因分析

目的分析颅脑损伤首次CT检查而复查CT、MRI阳性的表现及原因。方法对我科收治的78例颅脑损伤首次CT检查阴性，但复查CT、MRI发现迟发性颅内病变病人的临床资料进行回顾性分析。结果首次CT检查阴性但复查CT、MRI的阳性表现为迟发性血肿，脑干血肿，横窦沟硬膜外血肿，脑挫裂伤，蛛网膜下腔出血，硬膜下腔积液，弥漫性脑肿胀，弥漫性轴索损伤。首次CT检查阴性与检查时间，病变发展，受伤部位等因素有关。结论对首次头部CT检查阴性结果者,应仔细行神经系统的常规检查，密切观察患者的病情，及时复查CT、MRI以指导治疗，改善预后。     

千金藤碱对结肠癌细胞5-氟尿嘧啶耐药性的逆转作用和对HO-1和NQO1表达的影响

目的:探讨千金藤碱(Cepharanthine,CEP)逆转人结肠癌耐药细胞株Lovo/5-FU对5-氟尿嘧啶(5-fluorouracil,5-FU)的耐药性以及对HO-1和NQO1表达的影响.方法:采用药物浓度梯度递增法诱导建立结肠癌耐药细胞亚系LoVo/5-FU;用四甲基偶氮唑蓝(MTT)比色法检测千金藤碱和化疗药物对结肠癌细胞的细胞毒性;RT-PCR和Western blot技术检测NQO1和HO-1基因的mRNA和蛋白表达水平.结果:LoVo/5-FU细胞对包括5-FU在内的4种化疗药物产生耐药性,其中对5-FU耐药性最高,是LoVo细胞的18.82倍;千金藤碱对LoVo和LoVo/5-FU细胞增殖均有抑制作用,其浓度超过20-25 μg·mL-1时最为明显,但是对2种细胞的抑制效果无明显差别(P＞0.05);千金藤碱(2.5-10.0 μg·mL-1)可降低LoVo/5-FU细胞5-FU的IC50值(P＜0.05-0.01),在此浓度范围内,IC50值降低的程度与剂量成正相关;LoVo/5-FU细胞中HO-1和NQO1的mRNA和蛋白表达水平明显高于LoVo细胞(P＜0.05-0.01),10.0 μg·mL-1千金藤碱与5-FU联合处理后,LoVo/5-FU细胞中两个基因的表达水平明显降低(P ＜0.05-0.01).结论:千金藤碱可抑制结肠癌细胞增殖,增加结肠癌耐药细胞株Lovo/5-FU对5-FU的敏感性,原因可能与其抑制结肠癌细胞中HO-1和NQO1的表达水平有关.     

儿童急性淋巴细胞白血病化疗后合并结核性脑膜炎一例并文献复习

目的 探讨儿童急性淋巴细胞白血病(ALL)化疗后合并结核性脑膜炎(TBM)的临床特点及诊治.方法 回顾性分析福建医科大学附属协和医院2014年9月收治的1例ALL维持化疗中合并肺结核及TBM患儿的临床资料,并复习相关文献.结果 患儿,男性,11岁,确诊ALL 27个月,维持化疗中,出现中性粒细胞缺乏伴发热,予抗感染、抗真菌及甲泼尼龙治疗后,体温一度正常;1个月后再次发热,伴咳嗽、头痛,予抗感染、抗真菌治疗无好转,头痛加剧,并出现颈项强直,肺部CT示上肺及下肺背段炎症,脑脊液结核TB-DNA、结核分枝杆菌培养均阳性,确诊继发性肺结核、结核性脑膜炎,予规则抗结核治疗1.5年,病情治愈.结论 儿童ALL合并TBM的早期诊断是难点,及时、长疗程抗结核治疗,适当兼顾白血病化疗,仍可取得较好的抗结核病疗效,且不影响白血病预后.