Notch-2在甲状腺乳头状癌中的表达

目的:探讨Notch-2在甲状腺乳头状癌中的表达特点及其生物学意义。方法:应用RT-PCR法、免疫荧光法和Western-blot法,检测Notch-2人甲状腺乳头状癌与正常甲状腺组织标本中mRNA和蛋白的表达情况。结果:甲状腺乳头状癌组织中Notch-2 mRNA的表达明显低于正常甲状腺组织;免疫荧光法测定,Notch-2在正常甲状腺组织中表达呈强阳性,甲状腺乳头状癌组织中呈弱阳性;Western blot分析发现,Notch-2 mRNA在甲状腺乳头状癌中的表达低于正常甲状腺组织。Notch-2基因及蛋白表达在甲状腺乳头状癌组织和正常甲状腺组织之间明显不同。结论:在甲状腺乳头状癌组织中,Notch-2 mRNA及蛋白表达均下调,提示Notch-2信号通路受到普遍抑制,该基因在甲状腺癌发生、发展中起抑癌作用。     

DMBT1基因在上消化道癌组织中的表达及其意义

目的　探讨DMBT1基因转录表达异常在人上消化道癌组织发生、发展中的作用。方法　应用逆转录 聚合酶链反应 (RT PCR)检测 38例食管癌、2 1例贲门癌、2 5例胃癌肿瘤组织及其相应癌旁正常组织中DMBT1mRNA的表达缺失情况。结果　食管癌、贲门癌及胃癌组织中DMBT1mRNA阳性表达缺失率分别为 63 .2 % (2 4 / 38)、52 .4 % (1 1 / 2 1 )及 72 .0 % (1 8/ 2 5) ;伴有淋巴结转移的癌组织中DMBT1表达缺失率均显著高于相应淋巴结无转移的癌组织 (P <0 .0 2 5) ;肿瘤外侵越严重 ,DMBT1mRNA表达缺失率越高 (P <0 .0 2 5)。结论　DMBT1基因表达缺失与上消化道癌侵袭及淋巴结转移密切相关 ,提示该基因在上消化道癌的发生、发展及转移中起一定作用。     

LRIG2基因在膀胱癌中的表达及临床意义

目的:了解LRIG2基因及其产物在膀胱癌中的表达情况及其与肿瘤分级分期的关系。方法:采用逆转录-聚合酶链反应和Western-blot技术检测了38例膀胱癌组织和16例正常膀胱黏膜组织中LRIG2mRNA和蛋白的表达情况。结果:膀胱癌组织中LRIG2mRNA和蛋白的表达水平显著低于正常膀胱黏膜组织;且分级和分期越高,LRIG2mRNA和蛋白的表达水平越低。结论:LRIG2基因有可能是一种新的抑癌基因,其缺失或表达水平下调会导致肿瘤细胞生长分化,提示其有可能成为肿瘤基因治疗的又一靶点。     

抑癌候选基因NDRG2在结肠癌组织中的表达

目的分析抑癌候选基因NDRG2在人类结肠肿瘤组织中的表达情况。方法收集30例患者的结肠癌组织及其正常组织，提取总RNA，应用半定量反转录-聚合酶链反应（RT-PCR）检测NDRG2 mRNA的表达水平。为进一步分析NDRG2蛋白质的表达水平，分别提取30例组织的总蛋白，应用蛋白质印迹和免疫组织化学（免疫组化）技术检测其NDRG2的蛋白表达水平。结果RT-PCR结果显示，30例结肠癌组织中，有12例的NDRG2 mRNA表达明显降低。蛋白印迹结果显示，30例结肠肿瘤组织中有12例NDRG2蛋白水平明显下降，与RT-PCR结果一致。免疫组化结果显示，正常组织及肿瘤组织中NDRG2阳性率分别为90．0％（27／30）和53．3％（16／30），组间阳性表达率差异有统计学意义（P〈0．05），NDRG2的表达与患者的年龄、性别、淋巴结转移、浸润深度、Dukes分期无明显相关关系（P〉0．05），而与肿瘤组织分化级别相关，高、中分化组NDRG2阳性表达率高于低分化组（P〈0．05）。结论NDRG2在某些结肠癌组织中呈低表达，而且其表达与结肠癌的分化程度有关，提示其可能对结肠癌的发生或发展有重要的作用。这为研究结肠癌的发病机制提供了进一步的线索。     

Pin1和周期素D1在胰腺癌中的表达及其意义

目的:检测胰腺癌及癌旁组织中Pin1和周期素D1基因的表达,探讨Pin1在胰腺癌发病中所起的作用。方法:收集27例胰腺肿瘤组织及其相应的肿瘤旁组织标本,采用实时荧光定量逆转录聚合酶链反应法（RQ RT-PCR）检测胰腺良恶性肿瘤及肿瘤旁组织中Pin1和周期素D1 mRNA 的表达,运用Fisher精确概率分析两者之间的相关性及其与肿瘤临床分期和病理特征的关系。结果:7例胰腺囊腺瘤中周期素D1和Pin1的表达与肿瘤旁组织之间无显著性差异;而20例胰腺癌中周期素D1和Pin1的表达明显高于肿瘤旁组织［（2.78±1.02）vs.（4.36±1.27）和(5.48±1.69) vs. (9.97±1.86),P<0.05）］。Pin1和周期素D1与肿瘤的临床分期和病理分化程度无明显的相关（组间差异均为P>0.05）,但Pin1与周期素D1的表达有关（P<0.01）。 结论:胰腺癌中Pin1的过表达可促进周期素D1表达,由此诱导了肿瘤的发生;Pin1可能在胰腺癌中起着重要作用。     

二氢二醇脱氢酶2在前列腺癌中的表达及意义

目的探讨二氢二醇脱氢酶(DD)在前列腺癌中的表达及意义. 方法以β-actin为内参基因,应用逆转录聚合酶链反应方法检测DD亚型之一DD2在前列腺癌组织(11例)及正常前列腺组织(10例)中的表达水平.应用密度扫描技术定量分析RT-PCR产物电泳条带的密度. 结果 DD2 mRNA在前列腺癌组织中表达明显增高,吸光度比值0.550～1.018,中位数0.726;在正常前列腺组织中的表达水平较低,吸光度比值0.248～0.420,中位数0.333.癌与正常前列腺组织间DD2 mRNA表达差异有显著性意义(P<0.001). 结论 DD2 mRNA在前列腺癌中的高表达提示DD可能在前列腺癌的发生发展中发挥重要作用.     

神经生长相关蛋白-43基因及其蛋白在先天性巨结肠肠组织中的表达

目的探索神经生长相关蛋白-43(GAP-43)在先天性巨结肠痉挛段和扩张段肠组织中的表达情况,进一步探索先天性巨结肠的发病机理。方法收集2012年1月至2013年6月期间因先天性巨结肠于深圳市儿童医院行巨结肠根治术的30例患儿的活体肠组织标本,行免疫组化染色及荧光定量聚合酶链反应(RT-PCR)以检测扩张段和狭窄段肠组织中GAP-43 m RNA及其蛋白的表达,并比较扩张段和狭窄段肠组织中两者表达的差异。结果RT-PCR结果显示,痉挛组肠组织中GAP-43 m RNA的表达水平的中位数为0.052 8,低于扩张组的0.119 0(P<0.05)。免疫组化染色结果显示:30例患儿的狭窄段肠组织和扩张段肠组织的肌间神经丛及黏膜下神经丛中GAP-43蛋白均呈阳性表达,但痉挛段黏膜下神经丛和肌间神经丛的染色均较扩张段相应部位浅;与扩张组的黏膜下神经丛和肌间神经丛比较,痉挛组相应部位的GAP-43蛋白的平均光密度值均较低(P<0.05)。结论先天性巨结肠狭窄段肠组织中GAP-43蛋白的表达较扩张段肠组织下调,提示GAP-43蛋白可能是先天性巨结肠发病的危险因素之一。     

KiSS-1基因及其受体在乳腺癌中的表达及临床意义

目的 探讨人类转移抑制基因KiSS-1及其受体hOT7T175在乳腺癌中的表达情况及临床意义。方法 以逆转录-聚合酶链反应（RT-PCR）方法检测40例癌组织和癌旁正常组织中KiSS-1 mRNA和hOT7T175 mRNA的表达。结果 40例乳腺癌组织中，17例（42．5％）KiSS-1基因表达阳性，10例（25％）hOT7T175基因表达阳性；正常组织中29例（72．5％）KiSS-1基因表达阳性,20例（50％）hOT7T175基因表达阳性。KiSS-1基因和hOT7T175基因失表达与临床分期、肿瘤大小等均无显著相关性，而KiSS-1基因在淋巴结阴性者表达阳性率为63．6％，阳性者仅为16．7％。两者差异有统计学意义（P＜0．01）。结论 KiSS-1基因失表达与乳腺癌的淋巴结转移密切相关，提示该基因的表达可能是乳腺癌中一个重要的监控淋巴结转移的生物学标志。     

Prox-1在结直肠癌组织中的表达及意义

目的探讨结肠癌中Prox-1的表达与临床意义。方法选择2012年3月至2012年5月兰州军区兰州总医院肛肠科手术切除的结肠癌新鲜癌组织标本及癌旁正常组织标本41例,术前均未行放化疗,术后经病理证实,且有完整的临床、病理资料。采用RT-PCR法检测结肠癌患者肿瘤组织、正常组织中Prox-1的表达。结果结直肠癌组织中Prox-1的表达水平显著高于正常组织(P<0.05)。癌组织中Prox-1mRNA的表达水平与有无淋巴结转移组间比较差异有统计学意义(P<0.05)。Prox-1在癌组织的表达与临床分期之间存在正相关,且有统计学意义(P<0.05)。根据该结果可以认为临床分期越高,Prox-1表达越高。结论结直肠癌组织中Prox-1的高表达与结肠癌的淋巴结转移有一定关系,检测该指标对于了解结直肠癌的淋巴管生成与淋巴结转移的关系及判定预后具有一定价值。     

Survivin 基因在原发性胆囊癌中的表达及其意义

为探讨Survivin基因与原发性胆囊癌（PGC）发生、发展的关系。我们应用逆转录．聚合酶链反应（RT—PCR）和免疫组织化学法检测PGC组织中Sunrivin基因的表达情况，并将结果进行了相关分析。为原发性胆囊癌的综合治疗及基因治疗提供依据。     

肝部分切除后肝再生中Notch/Jagged信号及TNF和IL-1的表达

目的 研究Notch/Jagged信号传导通路及TNF和IL-1在大鼠肝部分切除术后肝再生中所起的作用.方法 取Wister大鼠行肝部分切除术,术后0,5,15,30 min和1,3,6,12 h及1,2,4,7 d留取再生肝组织,检测Notch-1、Jagged-1和增殖细胞核抗原(PCNA)蛋白的表达,RT-PCR检测Notch-1和Jagged-1的mRNA的表达,ELASA法测血清中TNF-α和IL-6的浓度.结果 肝部分切除后Notch-1蛋白在肝血窦内皮及门脉周围细胞表达增强,Jagged-1蛋白在胆管及门脉周围肝细胞上较强地表达.Noteh-1的mRNA表达量在6～48 h下调,Jagged-1的mRNA表达星在3～6 h上调,12～24 h下调.PCNA表达在术后12 h明显增加,1～2 d达到高峰.IL-6血清浓度在术后24 h达到高峰.TNF-α血清浓度高峰在3 h左右出现.结论 在肝脏被部分切除后,Notch/Jagged信号通路可以促进胆管的形成和结构维持,有助于新生血管的形成及肝细胞的增殖.TNF-α可以激活IL-6的表达,而IL-6参与肝细胞增殖过程中的启动、调控、终止等多个环节.     

先天性巨结肠症Wnt10b基因表达及其临床意义

目的 分析Wnt10b基因在先天性巨结肠症(HD)中的表达,探讨其在HD发生中的意义.方法 收集2003年至2010年中国医科大学附属盛京医院经病理检查证实的散发性HD狭窄段和正常段组织标本各60例.采用HE染色和免疫组织化学染色方法检测标本中Wnt10b的表达情况;再用荧光实时定量PCR(qRT-PCR)、Western blot从其基因和蛋白水平检测HD中Wnt10b的表达情况,并对其表达进行定量与比较分析.数据采用独立样本t检验.结果 经HE染色检测,60例患儿HD诊断明确,标本符合本研究要求.Wnt10b在HD狭窄段肠壁的肌间和黏膜下细胞胞质内呈强阳性反应,而在HD正常段肠壁的肌间和黏膜下细胞胞质内表达呈弱阳性或阴性;经免疫组织化学染色检测Wnt10b在HD狭窄段蛋白阳性面积百分率为0.061％±0.014％,正常段为0.006％±0.005％,两者比较,差异有统计学意义(t=2.955,P＜0.05).经qRT-PCR检测,Wnt10b在HD狭窄段肠管中mRNA相对含量为23.5±1.6,高于正常段肠管中的13.1±1.7(t=1.687,P＜0.05);60例患儿的HD狭窄段肠管组织标本中Wnt10b mRNA高表达者45例,低表达者15例.经Western blot检测,Wnt10b在HD狭窄段肠管中蛋白相对表达量为35.2±2.3,高于正常段肠管中的19.1±1.3(t =2.046,P＜0.05);60例患儿的HD狭窄段肠管组织标本中Wnt10b蛋白高表达者43例,低表达者17例.结论 Wnt10b mRNA与蛋白在HD肠管组织中的异常表达,与HD的发生有密切关系,可能在先天性消化道畸形的肠道发育中起一定作用.     

细胞黏附分子在先天性巨结肠症中的表达

目的　观察先天性巨结肠症 (HD)患者细胞黏附分子 (CAMs)在神经节正常肠段和神经节缺如肠段的表达情况。探讨CAM 成纤维细胞生长因子 (FGFR)信号传导在HD致病机制中的作用。方法　应用链酶抗生素蛋白 生物素 过氧化酶复合体法 (SABC) ,检测 16例HD患者中神经节正常和缺如肠段中CAMs的表达。结果　神经节细胞黏附分子 (NCAM )和神经元细胞钙黏素(N cadherin)在神经组织和平滑肌组织中都有表达 ,但在神经节正常肠段 (NG )肠组织中NCAM和N cadherin染色的神经纤维的数量明显多于神经节缺如肠段 (AG)。结论　CAMs在AG肠段的明显减少说明HD患者中CAM FGFR信号传导发生了异常 ,可能是导致肠神经母细胞移行障碍的原因之一。     

Reoperations in Hirschsprung disease

Background

We sought to identify causes of preventable complications related to operations for Hirschsprung disease.

Methods

We reviewed the cases of 51 patients with Hirschsprung disease who underwent a primary procedure elsewhere, had a complication, and were referred for reoperation.

Results

Thirty-five patients had 1 failed operation, 10 had 2, and 6 had 3. Initial operations were Soave (20), Duhamel (15), Swenson (5), transanal endorectal (4), myectomy (3), unknown (3), and laparoscopic Swenson (1). Thirty-one patients presented with a stoma. Patients without a stoma (20) had fecal impaction (8), recurrent enterocolitis (6), and fecal incontinence (6). None had both enterocolitis and incontinence. Reoperation was performed posterior sagittally (40) or transanally (5). Indications included stricture (21), megarectal Duhamel pouches (12), fistulae (11 [8 rectocutaneous, 2 rectourethral, and 1 rectovaginal]), pouchitis (2), and retained aganglionic bowel (8). After reoperation, 14 were continent, 11 had a stoma (8 permanent), 6 had voluntary bowel movements but soiled occasionally, 6 received rectal irrigations to avoid enterocolitis, 6 were incontinent but clean with bowel management, and 2 were lost to follow-up.

Conclusion

Stricture, megarectal pouch, fistula, and retained aganglionic bowel are preventable complications. Enterocolitis is partially preventable but can occur after a technically correct procedure. Fecal incontinence is a preventable complication likely because of anal canal damage.     

Advantages of intraoperative semiquantitative evaluation of myenteric nervous plexuses in patients with Hirschsprung disease

Background

The reappearance of an occlusive syndrome after surgical treatment of patients with Hirschsprung disease is often caused by incomplete resection of the affected segment. Intraoperative examination of frozen biopsies assists surgery, but interpretation of biopsies in the transitional zone is difficult.

Methods

We performed retrospective semiquantitative evaluations of myenteric nervous plexuses at the proximal limits of resection specimens from 41 children who were treated for rectocolic Hirschsprung disease using intraoperative examination of transparietal biopsies. The results of the evaluations were correlated with the postoperative courses of the children.

Results

After formalin fixation, the mean length of the aganglionic segments was 8.9 cm (range, 1-37.5 cm). At the proximal limit, the percentage of the circumference of the muscularis propria that was composed of plexuses with ganglion cells was more than 95% in 7 children, 90% to 95% in 21 children, 40% to 80% in 6 children, and 30% or less in 7 children (17%), 4 of whom subsequently developed occlusive syndromes.

Conclusions

Semiquantitative intraoperative examination of the entire circumference of resected segments facilitates precise evaluation of intrinsic innervation at the level of the anastomosis and may reduce the frequency of recurrence of occlusive symptoms after surgery.     

成人先天性巨结肠症的诊断和外科治疗

目的 探讨成人先天性巨结肠症的诊断和外科治疗。方法 回顾性分析1985年5月至2005年5月收治的成人先天性巨结肠症10例患者的临床资料。结果 男性7例，女性3例。年龄14．40岁。10例均有便秘史，均经钡灌肠检查确诊，5例肛门直肠测压均未引出直肠肛门抑制反射。无神经节细胞段在乙状结肠远端和直肠2例，直肠7例，未明1例。手术方法：乙状结肠造口1例，Ikeda法6例，Rehbein法2例，改良Swenson法1例。根治手术后排便功能：优7例，良1例，差1例。结论 成人先天性巨结肠症的诊断主要依据便秘史、钡灌肠检查和／或肛门直肠测压。拖出型手术是有效的手术治疗方法，手术方式应个性化。     

先天性巨结肠症和肛门直肠畸形中甲基化CpG结合蛋白2基因第3外显子突变分析

目的探讨甲基化CpG结合蛋白2（MeCP2）基因第3外显子突变与先天性巨结肠症（HSCR）和先天性肛门直肠畸形（ARM）的关系。方法采用PCR和DNA直接测序的方法．检测120例HSCR、50例ARM患儿和120名健康儿童外周血MeCP2基因第3外显子（MeCP2．E3）的突变情况。结果MeCP2．E3测序结果显示,120例HSCR患儿的碱基置换突变有45例（37．5％）．其中12例（10．0％）为突变型纯合子;50例ARM患儿的碱基置换突变有14例（28．0％）,其中4例（8％）为突变型纯合子：而健康对照儿童均未发现突变（P〈0．05）。结论HSCR和ARM患儿外周血MeCP2．E3存在突变．可能与疾病的发生有关．     

Clinical outcome in children after transanal 1-stage endorectal pull-through operation for Hirschsprung disease

Background/Purpose

Recently, the transanal 1-stage pull-through operation has been widely used in Hirschsprung disease (HD), and it is obviously superior to traditional approach in early term for its noninversion. However, the procedure is relatively so new that it makes assessment of the functional outcome and stooling patterns difficult. The aim of this study was to evaluate the clinical outcomes of the transanal 1-stage endorectal pull-through operation in the management of rectosigmoid HD.

Methods

Fifty-eight children (39 boys and 19 girls) aged 12 months to 13 years (mean, 2 years) who underwent transanal 1-stage endorectal pull-through operation for HD were followed up from 6 to 24 months. Clinical outcome was assessed by interviews and questionnaires. All patients had an aganglionic segment confined to the rectosigmoid area which was confirmed by the preoperative barium enema and postoperative pathological examination.

Results

Forty-six patients had satisfactory results without complications. In all the children, the mean stool times were 1 to 2 per day; only 4 had mean stool times of 8 to 10 per day. Postoperative soiling was present in 9, constipation in 5, and HD-associated enterocolitis in 3. There were no incontinence, cuff infection, anastomotic leak, and mortality in any of the patients. In the 12 symptomatic patients, there were 4 children with length of aganglionic segment less than 30 cm, and 8 had 30 cm or more. In the 46 asymptomatic patients, 42 had length of aganglionic segment less than 30 cm, and 4 had 30 cm or more. There was a significant difference between the group with less than 30 cm and the group with 30 cm or more of aganglionic segment. For statistical analysis, the Fisher exact test showed P < .05.

Conclusions

The transanal 1-stage endorectal pull-through is a feasible and safe procedure in children with rectosigmoid HD. The clinical outcome is satisfactory. A gradual recovery could be noted in the stooling patterns along with the time after surgery. The younger the patient operated on and the shorter the aganglionic segment, the lower do the stooling disorders occur and the faster does the stooling function recover.     

Stabilization period after 1-stage transanal endorectal pull-through operation for Hirschsprung disease

Purpose

The aim of this study was to evaluate postoperative outcomes and to find the period required for normal stooling pattern after the 1-stage transanal endorectal pull-through operation (TERPT).

Method

The authors retrospectively reviewed the clinical data and postoperative courses of 61 patients who had the aganglionic bowel confined to rectosigmoid and underwent TERPT between 2001 and 2007.

Results

Thirty-three patients (54.1%) were neonates, and 56 patients (91.8%) were less than 6 months old at operation. The mean age at TERPT was 90 ± 216 days, and the mean body weight at TERPT was 4.5 ± 2.8 kg. The average operating time was 189 ± 49 minutes, and mean length of bowel resection was 11.1 ± 3.2 cm. The mean postoperative hospital stay was 8.0 ± 3.6 days. Postoperatively, 5 (8.2%) patients were considered as failure of TERPT because of persistent problems in defecation. Fifty-six (91.8%) patients finally had normal stooling patterns and normal findings in abdominal radiography after 9.4 ± 6.2 weeks of the mean postoperative stabilization period. Neonatal cases had significantly longer postoperative stabilization periods than nonneonatal cases (11.3 ± 6.9 weeks vs 7.3 ± 4.6 weeks, P = .016). The postoperative stabilization period significantly decreased by age at operation as the patient's age increased (P = .018).

Conclusion

Clinical outcomes after TERPT are satisfactory, but a postoperative stabilization period is required for a normal stooling pattern to develop. The outcome of TERPT should consider a postoperative stabilization period.