IL-17在胆道闭锁患儿肝组织中的表达及意义

目的：研究胆道闭锁（Biliary Atresia BA）患者肝组织中白介素-17（IL-17）的表达水平及其与肝纤维化程度的关系。探讨其在BA发病和肝纤维变中的意义和可能的免疫作用机制。方法选用40例BA患者肝组织标本和10例非肝脏疾病死亡婴儿尸检肝组织标本,采用免疫组织化学法对肝组织进行IL-17染色,观察IL-17在两组肝组织中的表达。结果 BA组肝组织中IL-17表达水平平均秩次为30．28,对照组肝组织中IL-17表达水平平均秩次为6．4,经秩和检验,Uc＝4．773,P＜0.01,差异有显著统计学意义;BA组肝组织中IL-17表达水平与肝纤维化程度呈正相关（r＝0．8714,P＜0.01）。结论 IL-17在胆道闭锁发病过程及肝纤维化的发生发展中发挥了一定的作用。     

EpCAM阳性的不成熟胆道上皮细胞促进胆道闭锁肝纤维化的研究

目的 分析在胆道闭锁患儿肝组织中增生的EpCAM阳性的不成熟胆道上皮细胞与肝纤维化形成的关系,从而探讨不成熟的胆道上皮细胞在胆道闭锁病程进展中的作用.方法 选取2012年1月至2015年6月于我院就治的患儿肝石蜡标本进行连续切片,其中胆道闭锁(biliary atresia,BA) 65例,胆总管囊肿(choledochal cyst,CC) 27例,门静脉海绵样病变(cavernous transformation of portal vein,CTPV) 15例,应用CK19、EpCAM免疫组织化学染色及饱和苦味酸天狼星红染色方法,分析肝组织中增生的胆道上皮细胞的成熟性、EpCAM阳性的不成熟胆道上皮细胞与肝纤维化在位置上的关系,并应用Image-pro对400倍视野下肝组织切片中EpCAM阳性细胞进行计数,观察其增生数量与肝纤维化分级间的关系.结果 在CTPV组可见由CK19阳性的胆道上皮细胞组成的正常成熟的胆管,未见明显增生的胆道上皮细胞.在BA组及有纤维化的CC组可见胆道上皮细胞增生明显,邻近肝纤维化部位的胆道上皮细胞主要为EpCAM阳性的不成熟胆道上皮细胞,与肝纤维化存在位置上的关联.在胆道闭锁肝组织中随着肝纤维化的程度越高,EpCAM阳性的不成熟胆道上皮细胞的数量越多,且不同肝纤维化分期中EpCAM阳性细胞增生的数量存在显著差异(r=0.56,P＜0.001).结论 在小儿胆道闭锁疾病中,EpCAM阳性的不成熟胆道上皮细胞在短时间内迅速增生,可能是胆道闭锁患儿肝纤维化进展迅速的主要原因.     

胆道闭锁患儿肝组织细胞因子及相关基因表达的研究

目的 研究胆道闭锁(BA)患儿肝脏组织中上皮-间充质转化过程中特异性细胞因子的表达及胚胎期发育相关基因的表达激活.旨在探讨BA肝脏纤维化等系列改变的分子机制.方法 取经手术证实为BA患儿的8例肝活检组织(年龄2～3个月),以无消化道疾病尸检婴儿8例肝脏组织做正常对照(年龄0～2个月),RT-PCR半定量法检测胆管上皮细胞特异性因子CK19,纤维化早期指标Ⅰ型胶原蛋白COL1A1,发育相关基因Notch基因受体HES1,TGF-β及其阳性信号分子Smad3的表达水平变化.结果 BA患儿肝脏组织中CK-19,COL1A1、HES1、TGF-β及Smad3等基因表达水平均明显提高,实验组及对照组差异显著.结论 BA患儿肝脏组织胶原组织增生,同时有胆管上皮细胞增生,胚胎期肝脏组织发育相关基因Notch信号通路系统重新激活,TGF-β及其阳性信号分子Smad3在这一过程中发挥作用.     

CD4+T细胞因子在胆道闭锁患儿肝脏组织中的表达及其意义

目的 检测T淋巴细胞中的11种CD4+T细胞因子在胆道闭锁(biliary atresia,BA)患儿肝脏组织中的表达,以探讨其在胆道闭锁发病机制中的意义.方法 在病理的基础上,采用流式微球技术对29例BA及9例对照组患儿肝脏组织CD4+T细胞表达的11种细胞因子(IL-12p70、IFN-γ、IL-2、IL-10、IL-8、IL-6、IL-4、IL-5、IL-1β、TNF-α和TNF-β)同时进行定量检测,并对其代表因子IFN-γ进行免疫组化定位分析.结果 CD4+T细胞表达的11种细胞因子中,BA组患儿肝脏组织中IL-1β、IL-2、IL-6、IL-8、IFN-γ、TNF-α以及Th1细胞因子总量(IL-1β、IL-2、IL-8、IL-12p70、IFN-γ、TNF-α、TNF-β)与促炎因子总量(IL-1β、IL-2、IL-6、IL-8、IL-12p70、IFN-γ、TNF-α、TNF-β)分别为2920.69、1 106.01、152.22、12 614.22、834.18、161.29、19 504.55、19653.06,数值明显高于对照组的1 096.00、243.68、5.98、965.17、147.28、30.56、2 617.93、2 623.91,差异具有统计学意义(P＜0.05).结论 由CD4+Th1细胞及其细胞因子所介导的针对胆道上皮细胞的免疫炎症性疾病可能是造成胆道闭锁的主要原因之一.     

胆道闭锁患儿肝脏组织肥大细胞的表达及意义

目的 探讨胆道闭锁患儿肝脏组织肥大细胞表达及其与肝脏病变程度及预后的关系.方法 30例胆道闭锁患儿肝脏组织标本,肥大细胞采用甲苯胺兰染色和免疫组织化学染色.根据临床肝功能将患儿分为预后良好组与预后不良组,并取非消化系统疾病死亡的新生儿尸检肝脏标本作为对照组,比较各组肝组织肥大细胞表达情况.结果 胆道闭锁患儿肥大细胞数显著多于对照组(P<0.01),预后不良组肥大细胞数显著多于预后良好组(P<0.05).结论 肥大细胞参与胆道闭锁患儿肝脏损害、纤维化等病理过程,肥大细胞过度表达与胆道闭锁患儿肝脏的纤维化程度及预后密切相关.     

CD4+T细胞因子在胆道闭锁患儿肝脏组织中的表达及其意义

Objective To investigate the cytokines produced by CD4 + T cells in liver tissues of the patients with biliary atresia (BA). Methods The liver tissues of the 29 patients with BA were grounded and centrifuged. Then, the supernatants were collected. Nine patients not affected BA served as controls. The 11 cytokines produced by CD4 + T cells including IL- 12p70, IFN-γ, IL-2, IL- 10, IL-8,IL-6, IL-4, IL- 5, IL-1 β, TNF-α and TNF-β were detected using microsphere-based flow cytometry. Results Of the BA patients, the amount of IL-1β, IL-2, IL-6, IL-8, IFN-γ and TNF-α were 2 920. 69,1 106. 01,152. 22,12 614. 22,834. 18 and 161.29, respectively. The total amount of the Th1 cytokines including IL-1β, IL-2, IL-8, IL-12p70, IFN-γ, TNF-α and TNF-β was 19 504. The total amount of the proinflammatory cytokines including IL-1β, IL-2, IL-6, IL-8, IL-12p70, IFN-γ, TNFα and TNF-β was 19 653. 06. The amounts of these cytokines were significantly higher than those of the control group,which were 1 096.00,243.68,5.98,965.17,147. 28,30.56,2617.93, and 2 623.91, respectively. Conclusions The increased expressions of the cytokines expressed by the CD4 + Th1 cells may be one of the main causes of BA.     

胆道闭锁与病毒感染和免疫系统反应的相关研究

胆道闭锁的病因目前仍不清楚,了解其发病机制对于临床诊断及治疗具有至关重要的作用。近10年来,胆道闭锁可能是由病毒直接侵袭,或由病毒感染激发免疫异常所致的观点渐为小儿外科界所重视。病毒学研究发现,胆道闭锁可能与巨细胞病毒、轮状病毒或呼肠病毒等的感染有关。病理学研究提示,T细胞免疫反应很可能在胆道闭锁的发病中扮演了重要角色。该文总结了病毒学研究的现状,并从免疫调节基因、免疫细胞、炎症因子及自身免疫几方面阐述免疫系统反应研究的进展。     

Hedgehog通路在胆道闭锁肝纤维化中的研究进展

胆道闭锁(biliary atresia,BA)是一种罕见且严重的新生儿胆汁淤积症,其特征为进行性的毛细胆管纤维梗阻性疾病。近期研究发现,Hedgehog信号通路不仅在胚胎的形态形成、生长控制、肝胆发育中起着关键作用,还参与BA肌成纤维细胞的积累和肝纤维化的发生过程。本文通过检索文献,对Hedgehog信号通路的组成以...     

巨细胞病毒感染对胆道闭锁患儿预后影响的meta分析

目的 评价巨细胞病毒(Cytomegalovirus,CMV)感染对胆道闭锁患儿预后的影响.方法 全面检索Pubmed、Web of Science、中国知网和万方数据库,搜集有关巨细胞病毒感染对胆道闭锁患儿预后影响的相关文献,在全面文献回顾的基础上对文献进行筛选、评价和数据提取,并采用Stata12.0软件对纳入的文献进行Meta分析.结果 本研究共纳入7篇文献,Meta分析结果显示胆道闭锁患儿CMV感染组和CMV未感染组术后黄疸消退率差异有统计学意义(OR=0.23,95％CI=0.11～0.48,P＜0.01),其他2项指标的差异均无统计学意义.结论 现有证据表明,CMV未感染组术后黄疸消退情况较CMV感染组更好,巨细胞病毒感染能够对胆道闭锁患儿的预后产生一定的影响,但仍然需要更多的样本量,更加严密的分析,来进一步证实研究结论的可靠性.     

胆道闭锁患儿早期胆红素变化

目的：比较胆道闭锁及婴儿肝炎综合征（以下简称为婴肝）患儿出生后2个月内胆红素水平,分析胆道闭锁和婴肝患儿胆红素变化的规律,探讨胆道闭锁患儿胆红素水平与年龄的内在联系。方法选择2000年1月至2009年4月因黄疸于出生后60 d 内行血清胆红素检查的胆道闭锁患儿68例,为胆道闭锁组;2005年1月至2009年4月因婴儿肝炎综合征于出生后60 d 内行血清胆红素检查的患儿72例,为婴肝组;正常对照组为近1年内非肝脏疾病于出生60 d 内行胆红素检查的54例患儿。分析各组血清胆红素水平随年龄变化的特点,并计算检查时年龄、直接胆红素与总胆红素的比例。将上述结果分组进行对比分析,研究各组血清胆红素与年龄之间的关系。结果胆道闭锁患儿血清总胆红素、直接胆红素水平明显高于婴肝组患儿（P ＜0.05）。胆道闭锁组患儿出生后10 d 内血清总胆红素、直接胆红素水平明显高于婴肝组和正常对照组。胆道闭锁组患儿直接胆红素水平、直接胆红素与总胆红素比值随年龄增长缓慢升高,而婴肝组患儿直接胆红素水平、直接胆红素与总胆红素比值在出生20 d后变化甚微。结论胆道闭锁患儿总胆红素水平在新生儿期逐渐下降,1个月后再缓慢升高;且并不是一开始就表现为以直接胆红素升高为主的高胆红素血症,出生后20 d 内其与婴肝相似,均表现为以间接胆红素升高为主的黄疸。年龄小于2个月的婴儿,直接胆红素占总胆红素的水平超过0．7可以作为诊断胆道闭锁的线索。     

新生儿巨细胞病毒感染与胆道闭锁肝脏纤维化的相关研究

目的探讨新生儿巨细胞病毒感染与胆道闭锁肝脏损伤的关系。方法回顾我院2004年1月-2005年1月收治21例胆道闭锁患儿临床资料。对肝组织纤维化和肝细胞变性坏死程度分级，同时进行肝脏和肝门纤维块巨细胞病毒-pp65免疫荧光染色。根据血清及病毒学检查结果将病人分组，比较两组患儿肝功能，肝脏纤维化，肝细胞破坏程度。结果巨细胞病毒感染组13例，非感染组8例。两组肝功能各指标除总胆红素，γ-谷氨酰转肽酶外无明显差异。巨细胞病毒感染组肝脏纤维化程度重于非感染组（P〈0．05）。结论巨细胞病毒活动性感染加重胆道闭锁患儿胆汁淤积和肝脏的纤维化。     

Relationship between prognosis of biliary atresia and infection of cytomegalovirus

Background  The etiology of biliary atresia is still unknown. It is generally accepted that virus infection may be one of the important causes that lead to biliary atresia. This study aimed to illustrate the relationship between infection of cytomegalovirus and prognosis of biliary atresia. Methods  From January 2002 to March 2004, 27 patients who had undergone Kasai’s procedure because of biliary atresia were investigated for cytomegalovirus-IgG, IgM and pp65, and their mothers were also examined for confirmation of cytomegalovirus infection. The patients were divided into three groups: infection free group, cytomegalovirus positive group and cytomegalovirus infection group. The rate of jaundice disappearance and the incidence of reflux cholangitis were analyzed statistically. The histopathological changes of the liver were also analyzed. Results  The positive expression of cytomegalovirus-IgM and cytomegalovirus-pp65 in the patients was higher than that in their mothers (48% versus 14.81% and 37% versus 3.78%, respectively). Compared with the other two groups (80% in the infection free group, and 82% in the cytomegalovirus positive group), the rate of jaundice disappearance after operation in the cytomegalovirus infection group (36%) was significantly lower (P<0.05), and the incidence of reflux cholangitis was higher (P<0.05). Histopathological examination also showed that the degree of liver fibrosis and inflammation was more serious (P<0.05). Conclusions  There is a strong correlation between cytomegalovirus infection and a lower rate of jaundice disappearance, also a higher post-operational reflux cholangitis. Liver fibrosis seems to be more severe in biliary atresia patients with cytomegalovirus infection.     

Early cytomegalovirus infection and the long-term outcome of biliary atresia

Aim:  To study the impact of ongoing cytomegalovirus (CMV) infection at presentation of biliary atresia (BA) on the long-term outcome after Kasai procedure.
Methods:  Twenty-eight patients with BA born 1988–1997 were included and followed-up until 2007. Eleven patients (group A) had ongoing CMV infection at presentation and were compared to the remaining 17 patients (group B). Median age at Kasai procedure was 75 days in group A and 70 days in group B (p = 0.12).
Results:  Including all patients, survival with native liver was 50% and 36% at 4 and 10 years of follow-up respectively. At the end of follow-up, it was 25% and overall survival was 68%. When comparing groups A and B, neither difference in survival with native liver (p = 0.67, log-rank test) nor in survival after liver transplantation was detected.
Conclusion:  Survival with native liver after Kasai procedure is comparable to that of other centres. CMV positive patients may present with a later onset, alternatively the detection of CMV infection could delay the referral of BA patients. No significant differences in long-term outcome were detected with regard to early CMV infection.     

人巨细胞病毒感染与胆道闭锁CD4+T细胞因子的相关性研究

目的 检测胆道闭锁患儿肝脏组织中人巨细胞病毒滴度与11种CD4+T细胞因子表达水平之间的相关性,并探讨其意义.方法 分别采用荧光定量PCR和流式微球技术对29例胆道闭锁患儿肝脏组织中的巨细胞病毒滴度和CD4+T细胞表达的11种细胞因子(IL-12p70、IFN-γ、IL-2、IL-10、IL-8、IL-6、IL4、IL5、IL-1β、TNF-α和TNF-β)进行定量检测,并统计两者之间的相关系数.结果 人巨细胞病毒阳性者15例(51.7%),15例胆道闭锁患儿肝脏组织中人巨细胞病毒滴度与11种细胞因子具有不同程度的相关性,其中与部分细胞因子(IFN-γ、IL-2、IL-10、IL-4、TNF-α及TNF-β)呈强正相关,以IFN-γ的相关性最强(r=0.796),且该病毒的滴度与Th1类细胞因子总量(r=0.914)、促炎因子总量(r=0.913)及11种细胞因子总量(r=0.893)呈强正相关.结论 胆道闭锁可能是由人巨细胞病毒感染所促发,主要由CD4+Th1细胞及其细胞因子所介导的免疫炎症性疾病.

Abstract：

Objective To examine the relationship between human cytomegalovirus and CD4+ T cell's cytokines in biliary atresia. Methods HCMV titres and 11 cytokines (IL-12p70,IFN-γ,IL-2,IL-10, IL-8, IL-6, IL-4, IL- 5, IL- 1β, TNF-α and TNF-β) were assayed with Fluorescent quantitative PCR(FQ-PCR) and flowing microsphere technology respectively. The correlation coefficients were then calculated. Results Fifteen liver specimens(51.7%)were positive for HCMV. The titers of human cytomegalovirus showed different correlations with the 11 cytokines and strong positive correlation with some cytokines( IFN-γ、 IL-2、IL-1 0 、 IL-4 、 TNF-α and TNF-β), IFN-γ is the strongest one( r = 0. 796).Moreover, it shows strong positive correlation with the total of Th1 cytokines(r = 0. 914), pro-inflammatory cytokines(r = 0. 913)and the total of 11 cytokines(r= 0. 893). Conclusions The immune reaction of biliary atreisa may be triggered by the human cytomegalovirus, promoted by the CD4+ Th1 cells and their cytokines.     

巨细胞病毒感染与胆道闭锁Kasai手术预后的关系

目的探讨巨细胞病毒感染与胆道闭锁预后的关系。方法回顾性分析2013年1月至2014年8月我们收治的87例胆道闭锁病例,根据巨细胞病毒IgM抗体的测定结果分为阴性组55例和阳性组32例,分析术中肝活检组织的病理结果,AST与血小板比值指数(APRI),术后黄疸清除率和生存时间,比较其差异。结果术前两组年龄、总胆红素、ALT、AST和GCT比较,差异无统计学意义(P值均0.05)。术中肝活检病理结果根据Ohkuma Y分级,阴性组肝纤维化程度低于阳性组(P=0.002);两组APRI比较,术前差异无统计学意义(P=0.615),术后阳性组活动性肝纤维化占40.6%(13例),高于阴性组(11例,占20.0%),P=0.048;术后6个月阳性组黄疸清除率62.5%,阴性组78.2%,差异无统计学意义(P=0.139);两组累计生存率经log-rank检验比较,阴性组高于阳性组(P=0.034)。结论巨细胞病毒感染可能加重肝纤维化程度,影响胆道闭锁患儿的预后。     

Oesophageal atresia and biliary atresia: a rare association

Oesophageal atresia is known to be associated with cardiovascular, gastrointestinal, genitourinary and skeletal anomalies. Its association with biliary atresia is very rare and only a few cases have been reported in the literature. Although biliary atresia could be part of a multiple anomaly syndrome, its isolated occurrence is even rare. We present a case of oesophageal atresia associated with biliary atresia.     

Computerized three-dimensional study of a rotavirus model of biliary atresia: comparison with human biliary atresia

Biliary atresia is a panbiliary disease causing obstructive jaundice in neonates and infants. The clinical spectrum can be broadly categorized into the fetal and perinatal types. A consistent animal model that accurately mimics the whole clinical spectrum of biliary atresia is not yet available. However, rotavirus infection of neonatal mice has been shown to produce atresia in the biliary system. This study investigates the three-dimensional computerized morphology of the murine neonatal model comparing with age-matched control mice. Newborn Balb/c mice were injected intraperitoneally with rhesus rotavirus within 24–48 h after birth. Control mice received 0.9% NaCl. Pups with symptoms of cholestasis were sacrificed from the 5th to the 15th postinjection day, as were age-matched controls. Their hepatobiliary tissues were prepared for three-dimensional computerized image reconstruction. Rotavirus infection caused obliteration of the intrahepatic bile ducts and single to multiple atresias in the extrahepatic bile duct. At 15 days postinjection, intrahepatic ductal proliferation appeared, and the three-dimensional appearances of the intrahepatic biliary structures were similar to the human disease. Cystic duct and gallbladder dilatation was frequently seen in this model, and this feature distinguishes it from the human disease in which the gallbladder is almost always atretic. This rotavirus murine model demonstrates many of the features of human perinatal biliary atresia, and can be used as an investigative tool to further study the pathogenesis of biliary atresia.     

胆道闭锁胆汁成分与预后的关系研究

目的 探讨胆道闭锁术中胆汁成分变化与预后的关系。方法 本组对62例BA患儿在Kasai术中收集从肝门流出的胆汁,以25例胆总管囊肿患儿术中收集的胆汁作为对照组。采用全自动生化仪检测胆汁中总蛋白(TP)、胆汁酸(TBA)、总胆红素(TBIL)、直接胆红素(DBIL)、总胆固醇(CHO)、谷草转氨酶(AST)、谷丙转氨酶(ALT)、谷氨酰胺转肽酶(GGT)、碱性磷酸酶(ALP)的浓度。术后随访2～16个月,随访内容有黄疸消退时间,术后血生化,术后胆管炎发生情况及术后1年存活情况。结果 BA组胆汁中ALT(210.13±127.75) U/L、GGT(319.03±206.73) U/L、TP(45.86±18.16)g/L、CHO (0.08±(0.04) mmol/L浓度显著高于对照组,ALT:(69.39±44.55) U/L; GGT:(73.22±99.29)U/L; TP:(21.78±15.36)g/L; CHO:(0.02±0.03) mmol/L(P＜0.05); BA组胆汁中TBA( 189.81±107.81) μmol/L和DBIL (61.31±25.53 )μmol/L浓度显著低于对照组TBA:(735.06±199.75)μmol/L; DBIL:(90.43±27.18)μmol/L(P=0.000和P=0.035)。术前肝功能较好组胆汁中ALT(86.24±65.63) U/L、AST(801.64±516.50) U/L和GGT (205.82±112.25) U/L 浓度显著低于术前肝功能较差组,ALT:(244.67±175.69) U/L; AST:(1409.52±927.05) U/L;GGT:(362.05±235.57)U/L(P＜0.05);术前肝功能较好组胆汁中TBA (346.04±279.33)μmol/L 浓度显著高于术前肝功能较差组(192.47±93.55)μmbl/L(P=0.029)。术后3个月黄疸消退者胆汁TBA显著高于未消退者(251.18±131.83)μmol/L比(121.43±80.42) μmol/L,(P=0.007);术后3个月黄疸消退者胆汁DBIL(72.08±32.51 )μmol/L显著高于未消退者(40.22±23.47) μmol/L (P=0.022)。术后1年存活者胆汁TBA浓度较术后1年死亡者显著增高(258.63±105.63)μmol/L比(110.63±46.08) μmol/L,P= 0.015）;术后1年存活者胆汁ALT浓度较术后1年死亡者显著降低(8.72±8.83)U/L比(95.15±44.18)U/L,P=0.048。结论 BA术中胆汁成分与预后有明显的关系,胆汁中TBA较高、DBIL较高的BA患儿术后3个月黄疸消退较好;胆汁中TBA较高、ALT较低的BA患儿术后1年存活情况较好。胆汁TBA和ALT浓度可作为评价BA短期预后和中期预后的可靠指标。     

胆管闭锁与十二指肠乳头异位的关系探讨

目的 胆管闭锁的病因目前尚不清楚,本研究对胰胆的解剖进行造影研究.方法 对221 例胆管闭锁患儿中 47 例(21.3%)进行胆管造影,根据胆囊管和胆总管及十二指肠显影清晰影像进行研究,观察胆总管、胰管、胰胆合流共同管及其开口部位.以同期收治的 29 例非胆管闭锁性黄疸婴儿的胆管造影结果 为对照.结果 对照组29例中7例(24.1%)十二指肠乳头开口于十二指肠降段以远,而胆管闭锁组 47 例中 24 例(51.1%)十二指肠乳头开口于十二指肠降段以远,后者较前者明显增高(P<0.05).胆管闭锁组和对照组合并胰胆合流异常的发生率分别为95.7%(45/47)和82.8%(24/29),二者差异无显著性意义(P>0.122).胆管闭锁患儿胰胆合流共同管的平均长度指数为0.418±0.152,比对照组(0.236±0.122)明显延长,P<0.05.胆管闭锁组胆总管的平均直径指数为0.078±0.043,对照组为0.199±0.086,前者较后者明显细小,P<0.05,而共同管和胰管的直径差异不显著(P>0.05).结论 十二指肠乳头远端开口异位和胰胆合流共同管延长是胆管闭锁影像学改变特点,提示此畸形的病因与胚胎早期肝憩室发育异常有关.