毛囊干细胞定向分化过程中Wnt信号通路介导的基因调控

皮肤的毛囊干细胞具有自我复制以及多向分化潜能,在毛囊形态发育和定向分化过程中,Wnt信号通路起决定性作用。参与这条信号通路的重要蛋白质,如Wnt蛋白、Frizzled、B—catenin、GSK313、APC、Axin等研究相对较早,且颇为深入。但对于这条通路下游的调节因子,尤其是细胞核内关键性转录因子Tcf3、Lef1,以及它们所调控的一些重要基因c—myc、eyelinDl等的研究仍处于起步阶段。本文就Wnt信号通路介导的基因调节毛囊干细胞定向分化的研究现状进行综述．为构建组织工程皮肤提供理论参考。     

肝炎病毒感染与肝门部胆管癌发病关系的探讨

目的　探讨乙型肝炎 (乙肝 )和丙型肝炎 (丙肝 )病毒感染在肝门部胆管癌发病中的作用。　方法　采用免疫组织化学方法对 6 8例石蜡包埋肝门部胆管癌患者手术标本中乙肝病毒X蛋白和丙肝病毒C蛋白水平进行检测 ,并结合临床资料进行分析。　结果　 6 8例肝门部胆管癌中乙肝病毒X蛋白阳性率为 8 8% (6 / 6 8) ,丙肝病毒C蛋白阳性率为 35 3% (2 4 / 6 8) ,两者均阳性 1例(1 5 % ) ;乙肝、丙肝病毒感染的肝门部胆管癌在分化程度 (χ2 =8 7,P <0 0 1)、浸润程度 (χ2 =6 7 8,P<0 0 1)、淋巴结转移 (χ2 =4 3,P <0 0 5 )、根治程度 (χ2 =5 1,P <0 0 5 )与非病毒感染的肝门部胆管癌比较差异有显著意义。　结论　乙肝病毒X蛋白、丙肝病毒C蛋白可能在乙肝、丙肝病毒感染肝门部胆管癌的发病原因中起重要作用。乙肝、丙肝病毒感染的肝门部胆管癌恶性程度高 ,可能预后较差     

胆管癌组织螺杆菌16S-rRNA基因的检测及意义

目的探讨人胆管癌中是否存在螺杆菌的感染。方法取 11份手术切除的胆管癌标本 ,分别用两对螺杆菌属特异的 16S RNA引物进行PCR检测。对照组取自 4个肝囊肿和 2个血管瘤旁的正常肝胆管组织标本。结果 11份胆管癌标本用两对引物分别行二次PCR后 ,结果均为阳性的 6份 ,阳性率为 5 6 %。 4个肝囊肿和 2个肝血管瘤旁的正常肝胆管组织的PCR结果阴性 ,胆管癌组与对照组的阳性率有显著性差异 (P <0 0 5 )。对用C97和C98行PCR扩增的 2份产物测序 ,结果与幽门螺杆菌有 99%以上的同源性。结论胆管癌组织内存在螺杆菌的核酸物质 ,胆管癌患者肝脏中可能存在螺杆菌的感染。螺杆菌在胆管癌发生中的作用有待进一步研究。     

FXYD6在肝门胆管癌中的表达及临床意义

目的检测FXYD6蛋白在肝门胆管癌及相应癌旁远端正常胆管组织中的表达并分析其临床意义。方法应用免疫组织化学链酶亲和素-生物素-过氧化物酶复合物(SABC)法检测58例肝门胆管癌和30例远端正常胆管组织中FXYD6蛋白的表达,分析FXYD6蛋白表达与肝门胆管癌患者临床病理特征的关系。结果 FXYD6蛋白在肝门胆管癌组织中的表达阳性率明显高于其在癌旁远端正常肝门胆管组织中的表达阳性率,差异有统计学意义〔75.9%(44/58)比33.3%(10/30),χ~2=15.084,P=0.000〕。FXYD6蛋白在高、中分化肝门胆管癌组织中的表达阳性率明显高于其在低分化肝门胆管癌组织中的表达阳性率〔85.4%(35/41)比52.9%(9/17),χ~2=5.243,P=0.022〕,而FXYD6蛋白表达与肝门胆管癌患者的性别(χ~2=0.000,P=1.000)、年龄(χ~2=1.248,P=0.264)、T分期(χ~2=0.466,P=0.495)、淋巴结转移(χ~2=0.357,P=0.550)、病理分期(χ~2=0.005,P=0.944)及神经浸润(χ~2=3.016,P=0.082)均无关。结论 FXYD6在肝门胆管癌组织中的蛋白表达与其分化程度有关,其可能作为肝门胆管癌的一种新型肿瘤生物标志物。     

胆管癌组织中LRP/P170基因的表达及其临床意义

目的：探讨胆管癌组织中LRP和P170的基因表达情况及其临床意义。方法：收集手术切除的胆管癌标本21例作为实验组,另取8组胆管炎症标本作为对照组,采用RT-PCR方法检测标本中LRP及P170 mRNA的表达。结果：LRP及P170 mRNA在胆管癌组织中的表达阳性率分别76.2%和66.7%,显著高于对照组（P〈0.05）,上述指标与性别、年龄、病理分期、分化程度、淋巴结转移均无关（P〉0.05）。结论：LRP及P170在未经过化疗的胆管癌组织中均有不同程度的高表达;胆管癌的原发性多药耐药可能与LRP及P170有关。     

错配修复基因hMLH1,hMSH2在肝门部胆管癌中的表达和意义

目的 检测肝门部胆管癌中错配修复基因hMLH1,hMSH2失表达情况,研究其与肝门部胆管癌临床特征的关系,并探讨其与预后的相关性.方法 用免疫组织化学技术检测54例肝门部胆管癌组织、25例正常胆管中hMLH1,hMSH2表达,结合临床病理学资料及随访统计资料进行统计学处理分析.结果 (1)在54例肝门部胆管癌组织中,hMLH1蛋白阳性表达24例,其阳性率44.4%;25例正常胆管组织中,hMLH1蛋白阳性表达23例,其阳性率92.0%;hMSH2蛋白癌组织阳性表达21例,其阳性率38.9%;hMSH2蛋白在正常胆管组织阳性表达21例,其阳性率84.0%.hMLH1,hMSH2在肝门部胆管癌组织中的表达明显减少,二者比较差异具有统计学意义(P＜0.05).(2)根据统计学检验,hMLH1,hMSH2表达与肝门部胆管癌Bismuth分型(P＞0.05)、病人性别(P＞0.05)、年龄(P＞0.05)、肿瘤大小(P＞0.05)无关,但与病理分级(P＜0.05)和淋巴结转移(P＜0.05)具有显著相关性.(3)hMLH1表达阴性组术后2年生存率明显低于表达阳性组(15%VS 45.4%,P＜0.05).hMSH2表达阴性组术后2年生存率明显低于表达阳性组(23.5%VS44%,P＜0.05).结论 hMLH1,hMSH2在肝门胆管癌中的失表达在肿瘤的发生、发展及转移中起重要作用,是判断预后有价值的指标.     

肝门部胆管癌的临床诊断与手术治疗分析

目的探讨肝门部胆管癌的临床诊断方法及两种不同手术方法的预后。方法选取2002年12月至2008年3月就诊的肝门部胆管癌患者43例作为研究对象,回顾性分析所有患者的临床表现、影像学检查结果、手术方式、生存率等临床资料,所有对象按手术方法分为根治性切除术组（22例）和姑息性切除术组（21例）,对比分析两组间患者的术后并发症发生率,1、3、5年生存率有无统计学差异。结果临床表现以黄疸最为常见,占86．0％,其次是尿黄（81．4％）和皮肤瘙痒（72．1％）;经过常规超声、CT及MRI联合检查,根治性切除术组和姑息性切除术组对肝门部胆管癌检出率分别为90．9％和95．2％;MRI诊断阳性率明显高于超声诊断阳性率（χ^2=7．379,P〈0．01）。根治性切除术组患者并发症发生率明显高于姑息性切除术组（χ^2=14．321,P〈0．01）,1、3、5年生存率也明显高于姑息性切除术组（r=6．018、χ^2=2．842、r=17．483,P〈0．05）。结论对于肝门部胆管癌患者,MRI诊断阳性率明显高于超声,采取超声联合CT或MRI可提高早期诊断率;根治性切除术治疗肝门部胆管癌,能够提高此类患者的远期存活率。     

伴神经浸润的肝门部胆管癌的病理和临床分析

目的总结伴神经浸润的肝门部胆管癌的临床病理类型、手术方式,探讨伴神经浸润的肝门部胆管癌治疗方法及远期存活率。方法对1993年1月至2004年12月收治的311例肝门部胆管癌(神经浸润组89例、无神经浸润组222例)的临床病理及随访资料进行回顾性分析。结果神经浸润是肝门部胆管癌的一种重要转移方式。伴神经浸润组的89例肝门部胆管癌的1、3、5年存活率分别为65.59%、22.71%、14.83%,无神经浸润组的222例肝门部胆管癌的1、3、5年存活率分别为80.62%、35.14%、24.98%,无神经浸润组存活率明显高于神经浸润组(P=0.037)。结论肿瘤的分化程度、手术方式是影响预后的重要因素。根治性切除是目前较理想的治疗方式。伴神经浸润的肝门部胆管癌患者的预后较差,远期疗效仍无显著改善,根治性切除术可望改善其预后。     

趋化因子受体CCR7诱导血管生成拟态形成在肝门部胆管癌中的作用

目的:探讨趋化因子受体CCR7和血管生成拟态(VM)在肝门部胆管癌组织的表达及其与临床病理因素的关系。方法:免疫组织化学染色检测63例肝门部胆管癌标本和20例正常胆管组织CCR7的表达,PAS双重染色检测血管生成拟态的表达,将CCR7表达和VM形成与临床病理因素进行相关性分析。结果:63例肝门部胆管癌组织中,CCR7阳性率79.4%。正常胆管表达率10%;胆管癌组织25例存在血管生成拟态形成,正常组织中无Vm形成;与临床病例因素相关性分析,CCR7和VM在肝门部胆管癌中的表达与肿瘤的分化程度和淋巴结转移呈显著相关;单变量和多变量相关性分析CCR7、VM、分化程度和淋巴结转移与生存率有关,而CCR7是影响肝门部胆管癌生存率的独立性因素。结论:肝门部胆管癌存在CCR7和血管生成拟态的表达,两者呈正相关;CCR7和VM的表达与肝门部胆管癌的淋巴结转移和肿瘤分化程度有关;CCR7是影响肝门部胆管癌生存率的独立性因素。     

肝门部胆管癌中乙肝、丙肝病毒感染与p53异常表达的关系及意义

目的 探讨乙肝病毒，丙肝病毒感染在肝门部胆管癌发病中的作用及与p53异常表达的关系。方法 采用免疫组织化学方法对68例石蜡包埋肝门部胆管癌标本中的HBV－X，HCV－C和p53蛋白进行检测，并结合临床资料进行分析。结果 68例肝门部胆管癌中乙肝病毒X蛋白阳性率为8．8％（6／68），丙肝病毒C蛋白阳性率为35％（24／68），两者均阳性1例（1．5％）；p53蛋白阳性率为45．6％（31／68）。乙肝，丙肝病毒感染的肝门部胆管癌在分化和浸润程度，淋巴结转移，根治程度与非病毒感染的肝门部胆管癌有明显差异，p53与乙肝病毒X，丙肝病毒C蛋白表达呈显著的正相关（P＜0．001）。结论 乙肝，丙肝病毒感染与肝门部胆管癌发生有关。乙肝，丙肝病毒感染的肝门部胆管癌恶性程度高，可能有较差的预后。HBV－X，HCV－C蛋白可能在HBV，HCV感染肝门部胆管癌的病因中起重要作用，p53蛋白在肝门部胆管癌中的异常表达与HBV，HCV感染有关。     

凋亡相关基因bcl-2和bax在胆管癌组织中表达的意义

目的　探讨细胞凋亡抑制基因ｂｃｌ－ ２ 和凋亡促进基因ｂａｘ 在胆管癌组织中的表达。方法　应用免疫组化方法对２０ 例胆管癌（ＣＨＣ）和７例先天性胆总管囊肿（ＣＣＣ）组织中ｂｃｌ－ ２和ｂａｘ 蛋白进行检测。结果　２０例ＣＨＣ中有１例ｂｃｌ－ ２ 蛋白表达阳性（５％ ）,７例ＣＣＣ中ｂｃｌ－ ２均表达阴性。２０例ＣＨＣ中有１１ 例ｂａｘ 蛋白表达阳性（５５％ ）,７例ＣＣＣ中仅１ 例表达阳性（１４．２９％ ）,两组间差异有显著意义（Ｐ＜ ０．０１）。结论　ｂｃｌ－ ２ 和ｂａｘ 蛋白的改变在胆管癌发生发展过程中不起重要作用。     

Reduced Axin Protein Expression Is Associated with a Poor Prognosis in Patients with Squamous Cell Carcinoma of Esophagus

Aims  Our study investigates the significance of the expression of Wnt pathway proteins including β-catenin, Axin, β-transducin-repeat-containing protein (β-TrCP), and adenomatous polyposis coli (APC) in squamous cell carcinoma of the esophagus (ESCC). Methods  Immunohistochemical analysis was performed on paraffin-embedded tissue specimens from 128 resected ESCC tumors to detect the expression of β-catenin, Axin, β-TrCP, and APC. Correlation between immunoexpression, clinicopathological parameters, and patient survival was analyzed. Results  Increased β-catenin expression was noted in 22 (18.2%) of 121 tumor specimens. Reduced expression of Axin, β-TrCP, and APC was observed in 57 (46.0%) of 124, 29 (24.4%) of 119, and 54 (48.2%) of 119 specimens, respectively. No correlation was found among these protein expressions. Axin protein expression was inversely correlated with tumor invasion depth (P = 0.033). Reduced Axin protein expression, lymph node involvement, and distant metastasis were significant negative predictors for overall survival and disease-free survival on univariate analysis. In multivariate analysis, reduced Axin expression remained a significant prognostic factor for patients with ESCC (P = 0.005). Conclusions  Reduced Axin expression was observed in 46% of ESCC tumor specimens and was associated with poor prognosis in patients with ESCC. Further study is mandatory to elucidate the underlying mechanism responsible for loss of Axin expression and the role of Axin in ESCC tumorigenesis. Anna Fen-Yau Li, Po-Kuei Hsu, and Ching Tzao contributed equally to this article.     

Concomitant hilar cholangiocarcinoma with choledochal cyst and cholelithiasis in an asymptomatic patient: A case report

Introduction and importanceCholedochal cyst is an important risk factor for cholangiocarcinoma. Concomitant hilar cholangiocarcinoma with choledochal cyst with cholelithiasis is a rare finding.Case presentationA 52-year-old male presented with non-specific complaints of generalized weakness for 3 months. Transabdominal ultrasound showed dilated common bile duct with hyperechoic mass at hilar region; further evaluation with magnetic resonance cholangiopancreatography and contrast-enhanced computer tomography of abdomen and pelvis revealed concomitant intraductal hilar cholangiocarcinoma (Bismuth Corlette type 1) with Choledochal cyst (type IVa) with cholelithiasis. After optimization patient underwent left hepatectomy with common bile duct excision with cholecystectomy with Roux-en-Y hepaticojejunostomy. Histopathological examination confirmed it to be well-differentiated adenocarcinoma, intestinal type at the hilar confluence.Clinical discussionAn asymptomatic male patient with the concomitant finding of perihilar cholangiocarcinoma (Bismuth Corlette type I) with choledochal cyst type IVa with cholelithiasis is a rare finding. The patient was managed with surgical excision of the common bile duct with Roux-en-Y hepaticojejunostomy and cholecystectomy. Diagnostic evaluation should be proper not to miss or overlook such a synchronous lesion.ConclusionIncidental finding of concomitant perihilar cholangiocarcinoma with the choledochal cyst is rare. Proper evaluation of the patient with CECT abdomen and pelvis and MRCP is necessary for the diagnosis. Proper surgical resection with adequate lymph node removal is important for surgical clearance.     

LRP5 mutations in osteoporosis-pseudoglioma syndrome and high-bone-mass disorders

The LDL receptor-related protein 5 (LRP5) is a member of the LDL receptor family, which also includes the VLDL receptor and the apolipoprotein E receptor 2. The LRP5 is a co-receptor of Wnt located on the osteoblast membrane between two other receptors, Frizzled and Kremen. Frizzled and LRP5 bind to Wnt, thereby stabilizing beta-catenin and activating bone formation. When the dickkopf protein (Dkk) binds to Kremen and LRP5, this last undergoes internalization and therefore becomes unable to bind Wnt; this leads to degradation of beta-catenin and to inhibition of bone formation. In humans, loss of LRP5 function causes osteoporosis-pseudoglioma syndrome, which is characterized by congenital blindness and extremely severe childhood-onset osteoporosis (lumbar spine Z-score often < -4) with fractures. The G171V mutation prevents Dkk from binding to LRP5, thereby increasing LRP5 function; the result is high bone mass due to uncoupling of bone formation and resorption. The Z-scores in this condition can exceed +6 at the hip and spine. The LRP5 and Wnt/beta-catenin reflect the level of bone formation and play a central role in bone mass accrual and normal distribution. Furthermore, LRP5 may contribute to mediate mechanical loads within bone tissue. Identification of the Wnt/beta-catenin pathway is a breakthrough in the elucidation of pathophysiological mechanisms affecting bone tissue and suggests new treatment targets for patients with osteoporosis or specific malignant conditions such as myeloma and sclerotic bone metastases.     

先天性胆总管囊肿再次手术的原因分析及治疗

目的 探讨先天性胆总管囊肿再次手术的原因及治疗方法.方法 对41例先天性胆总管囊肿再次手术患者的临床资料进行总结分析.结果 按先天性胆总管囊肿Todani分型,41例中Ⅰ型32例,Ⅱ型1例,Ⅳ型3例,Ⅴ型5例(其中1例呈弥漫性左右肝分布).既往有1次胆道手术史者30例,2次者8例,3次者3例.41例中合并有胆管结石40例,胆道感染33例,肝门部胆管狭窄7例,原胆肠吻合口狭窄6例,囊肿癌变5例.本次行切除肝外胆管囊肿、肝管空肠Boux-en-Y吻合术者32例,其中2例囊肿癌变者同时行肝十二指肠韧带骨骼化和区域淋巴结清扫术;行肝叶切除联合胆总管囊肿切除、肝管空肠Roux-en-Y吻合术者8例;行胆肠吻合口狭窄切开成形、胆肠内引流重建术1例.41例术后均获随访,随访时间3个月至10年,平均53.7±32.1个月,随访期内发生胆道感染4例,无吻合口狭窄或结石复发等并发症发生.结论 胆总管囊肿术式选择不当及诊断失误导致的多种并发症是导致再手术的主要原因,肝内外胆管囊肿切除、肝管空肠Roux-en-Y吻合术可取得较为满意的疗效.     

Intrahepatic cholangiocarcinoma arising 33 years after excision of a choledochal cyst: report of a case

We report a case of intrahepatic cholangiocarcinoma arising 33 years after excision of a choledochal cyst. A 61-year-old woman was admitted to our hospital complaining of fever. Thirty-three years ago she had undergone extrahepatic choledochal cystectomy and choledochojejunostomy for a choledochal cyst. Computed tomography showed a tumor in the anterior segment of the liver, extending to the posterior and medial segments and the right portal vein. Intrahepatic biliary stones were seen in the bile ducts. We performed extended right lobectomy. Microscopically, the tumor was cholangiocarcinoma. Most of the tumor area was composed of invasive adenocarcinoma but a carcinoma-in-situ component was also observed in some regions including the hilar bile duct, where an intrahepatic biliary stone was seen. This suggests that the cancer development could be related to intrahepatic cholestasis. Patients with choledochal cyst may have to be carefully followed up for more than 30 years even after diversion surgery.     

Wnt信号途径在不同性质乳腺组织和细胞中的表达与分布

目的 探讨乳腺癌发生发展中Wnt信号途径的改变及其关机制.方法 用基因芯片和荧光实时定量聚合酶链反应(PCR)技术检测5例乳腺癌患者不同性质乳腺组织和细胞中Wnt信号途径相关基因的表达.结果 与正常乳腺组织比较,癌组织中Frizzled 3、Lrp5、Lrp6、Wnt9a、Wnt10a表达分别上调了8.11、10.28、9.37、15.47、19.63倍(发生频率均≥60%),DDK1、DDK4、WIF1表达分别下调了10.87、9.62、12.05倍(发生频率均≥60%);其中Wnt9a、Wnt10a以及DDK1、DDK4、WIF1表达的改变主要在癌源性成纤维细胞,分别上调40.88、61.59倍和下调38.46、19.61、29.41倍(P<0.01);Frizzled 3、Lrp5、Lrp6表达改变主要在乳腺癌千细胞,分别上调了27.36、31.41、21.25倍(P<0.01).结论 在乳腺癌发生发展过程中,成纤维细胞源性因子Wntga、Wnt10a和DDK1、DDK4、WIF1的改变参与了Wnt信号途径的异常,并主要影响乳腺癌干细胞的自我更新调控.