HIF-1α和VEGF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)患儿肝组织中缺氧诱导因子(HIF-1α)和血管内皮细胞生长因子(VEGF)的表达及与血管生成和纤维化的关系,探讨其在BA发病中可能的作用机制。方法选取胆道闭锁患儿(BA组)肝组织15例、胆管扩张症患儿(CBD组)肝组织10例、BA发展为肝硬化接受肝移植患儿(LT组)肝组织10例。HE、Vimentin染色观察各标本的组织学改变并判定纤维化程度,CD34染色标记血管计数微血管密度(MVD),免疫组化法检测HIF-1α和VEGF在各组肝组织中的表达。结果 CBD组、BA组、LT组微血管密度分别为16.8±1.9、23.2±3.8、25.9±2.1。HIF-1α在肝细胞胞浆和部分胞核中表达,BA组HIF1-α的光密度(0.160 7±0.029 7)高于CBD组(0.150 4±0.146 3)和LT组(0.151 3±0.005 9),P0.05。VEGF在肝细胞、胆管、动脉壁的表达,BA组VEGF的光密度(0.162 9±0.012 9)高于CBD组(0.132 2±0.025 7)和LT组(0.146 2±0.015 2),P0.05。结论 HIF-1α、VEGF可能通过参与BA中的血管生成过程,促进肝纤维化。     

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.     

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。     

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...     

Hedgehog信号通路在胆道闭锁肝纤维化中的表达及机制研究

目的:探讨Hedgehog信号通路和上皮-间充质转化(epithelial-mesenchymal transition, EMT)在胆道闭锁(biliary atresia, BA)肝纤维化进展中的作用,为BA肝纤维化发生机制提供实验依据。方法:选取2019年1月至2020年1月天津市儿童医院普外科12例BA患儿肝组...     

PDCD5在胆道闭锁患儿胆管组织中的表达及意义

目的研究促凋亡基因PDCD5在胆道闭锁及胆总管囊肿患儿胆管组织中的表达规律,探讨促凋亡基因PDCD5在胆道闭锁发病机制中的作用。方法分别取18例胆道闭锁（Biliary Atresia,BA）和11例胆总管囊肿（Choledochal cyst,CC）患儿的胆管组织,应用蛋白质印迹（westong blot）与荧光实时定量（quantitative real—time PCR,qRT-PCR）的方法分别检测PDCD5在胆道闭锁与胆总管囊肿患儿胆管组织中的表达,并进行定位和定量分析与比较。结果PDCD5在CC组胆管组织中的CT值为（6．7292±1．169）,高于BA组的相应CT值（4．0125±1．0735）,P〈0．05。应用2-△△CT计算出CC与BA中PDCD5基因表达量的比值为1：6．57。PDCD5在BA组胆管组织中mRNA转录水平明显高于对照组;Western blot结果显示,PDCD5在BA组胆管组织中蛋白表达量明显高于CC组。结论PDCD5在胆道闭锁患儿胆管组织中的蛋白和基因表达水平均增强。PDCD5可能参与胆道闭锁胆管上皮细胞凋亡,从而参与胆道闭锁的炎性反应,在胆道闭锁的发病机制中发挥重要作用。     

MicroRNA在胆道闭锁中的研究进展

小分子核糖核酸(microRNA,miRNA,miR)是一类约22个核苷酸的单链非编码RNA,在转录后水平调节基因的表达.miRNA广泛存在于人体各个组织器官中,在细胞增殖、分化、凋亡、胚胎发育、信号传导、肿瘤发生及代谢等过程中发挥着重要作用.近年来研究表明,某些miR-NA表达的改变在胆道闭锁的发生发展中起着重要作用,在该过程中体现出极其复杂的调控机制,同时miRNA的研究也为胆道闭锁的早期诊断和治疗带来了新的希望和思路.本文就miRNA在胆道闭锁中的研究进展做一综述.     

胆道闭锁肝纤维化病变中平滑肌肌动蛋白表达的研究

目的 本研究通过检测平滑肌肌动蛋白(α-SMA)在胆道闭锁(BA)肝组织和肝外胆系的表达,探讨肝纤维化过程与临床预后的关系.方法 采用免疫组化染色方法对2005年7月至2006年5月本院21例BA肝组织、肝门纤维块进行CD68和α-SMA染色;对照组为5例胆汁淤积和10例胆总管囊肿.选用LEICA-DM研究级生物显微镜,QWIN软件环境下测量抗体阳性细胞面积百分比和平均光密度,随访19例BA术后3个月直接胆红素下降比率.同时对21例BA肝组织纤维化分级,并与α-SMA表达量进行相关分析.结果 α-SMA在BA肝纤维块高度表达于胆管上皮、胆管周围的胶原纤维;α-SMA阳性表达量和表达强度明显高于对照组,其表达量与表达强度呈线性正相关(r=0.549,P=0.022);CD68在BA肝组织表达较对照组明显增强,但肝门纤维块鲜有表达.α-SMA表达量与肝脏纤维化分级呈正相关(P=0.02);α-SMA阳性表达面积百分比与术后3个月直接胆红素下降率呈负相关(r=-0.653,P=0.029),但肝组织CD68表达与α-SMA阳性表达相关性不明显(r=0.444,P=0.057).结论 α-SMA的表达可能是肝脏纤维化的早期标志,SMA的阳性表达量与术后3个月胆红素消退呈显著负相关,预示BA肝内外胆管系统纤维化,乃至肝硬化,提示可能临床预后不佳.     

血清半乳糖凝集素-3水平判断胆道闭锁患儿术后预后的意义

目的 探讨半乳糖凝集素-3(Gal-3)、透明质酸(HA)与胆道闭锁(BA)患儿术后肝损害、肝纤维化的关系.方法 选择行Kasai术术后BA患儿为BA组(n=35),同期体检健康儿童为健康对照组(n=10).BA组根据胆红素水平分为黄疸组(n=11)和无黄疸组(n=24),根据胃镜下食管静脉曲张情况分为门静脉高压(PH)组(n=27)和非PH组(n=8).应用自动生化仪检测2组肝功能,采用ELISA检测2组血清Gal-3水平,采用放射免疫法检测2组血清HA水平.结果 BA组肝损害、肝纤维化程度较健康对照组重,血清Gal-3、HA水平均较健康对照组显著升高(Pa＜0.05);黄疸组肝损害、肝纤维化程度较无黄疸组重,血清Gal-3、HA水平均较无黄疸组显著升高(Pa＜0.05);无黄疸组血清Gal-3水平较健康对照组显著升高(P ＜0.05);PH组血清Gal-3水平较非PH组显著升高(P＜0.05);血清Gal-3水平与总胆红素、ALT、γ-谷氨酸转肽酶水平呈正相关(Pa＜0.05),与HA水平亦星正相关(P＜0.05).结论 血清Gal-3水平与BA术后患儿肝损害程度和肝纤维化程度密切相关,可作为BA术后判断患儿预后的重要生化指标.     

CCL25/CCR9在胆道闭锁肝纤维化中的表达及临床意义北大核心CSCD

目的探讨CC趋化因子受体9(CC chemokine receptor 9,CCR9)及其趋化因子配体25(CC chemokine ligand 25,CCL25)在胆道闭锁(biliary atresia,BA)患儿肝纤维化中的表达并评价其临床价值。方法回顾性分析2019年10月至2022年8月在天津市儿童医院普外科进行肝脏手术治疗的35例患儿临床资料,将诊断为胆总管囊肿(choledochal cyst,CC)的8例患儿作为对照组,其余27例BA患儿为BA组。所有患儿手术均留取肝组织样本,同时收集患儿血清样本,采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清CCL25水平,并绘制受试者操作特征(receiver operator characteristic,ROC)曲线,结合临床生化指标进一步分析血清CCL25对BA患儿的辅助诊断价值。采用免疫组织化学染色和实时荧光定量聚合酶链反应(real time fluorescent quantitative polymerase chain reaction,qRT-PCR)检测肝组织中CCR9的表达水平,并采用Spearman相关分析法分析其与BA患儿肝组织纤维化分级的相关性;分析CCR9对BA患儿肝脏纤维化程度的评估价值。计数资料组间比较用χ^(2)检验或Fisher精确检验;不符合正态分布的计量资料组间比较采用Mann-Whitney U检验和Wilcoxon检验。结果ELISA结果显示,BA组血清CCL25水平310.03(262.03,338.87)pg/ml明显高于CC组244.46(222.70,260.45)pg/ml,差异有统计学意义(P=0.004)。ROC曲线显示,血清CCL25水平诊断BA患儿的曲线下面积为0.829(95%CI:0.683~0.975),临界值为267.12 pg/ml。血清CCL25水平、GGT、TBA联合诊断BA的曲线下面积为0.958。免疫组织化学和qRT-PCR结果显示,BA患儿肝组织中CCR9蛋白表达量[40.29(29.40,65.12)比15.26(8.04,20.15),P<0.001]、CCR9 mRNA表达量[3.88(1.38,7.95)比0.83(0.07,0.95),P=0.0005]均明显高于CC组。与Ⅰ~Ⅱ级肝纤维化比较,Ⅲ~Ⅳ级肝纤维化的CCR9蛋白表达量[51.08(39.36,73.27)比25.93(19.23,38.00),P=0.0022]、CCR9 mRNA表达量[6.63(2.75,9.45)比1.21(1.04,3.57),P=0.0075]明显提高。BA组患儿的CCR9蛋白表达量、mRNA表达量与肝纤维化分级均呈正相关(rs=0.820、rs=0.804,P<0.001)。结论血清CCL25在辅助诊断BA患儿方面具有潜在的临床价值;CCR9在肝组织中的表达与BA患儿肝纤维化程度密切相关,且随着肝纤维化程度加重而增加。     

Evaluation of catch-up growth after liver transplantation in children with biliary atresia

Orthotopic liver transplantation (Tx) has improved survival in infants with extrahepatic biliary atresia (BA) when portoenteroanastomosis fails. Symptoms leading to Tx include liver failure, poor quality of life and growth failure. The objective of the study was to determine catch-up growth in children with BA. Medical records and growth data of 36 patients (24 girls) who received a Tx due to BA were analyzed. Thirty-two patients completed 3 yr and 15 patients 7 yr of follow-up after Tx. At Tx, the median age was 2.7 yr (range 0.7-12.6) and mean height Z score (+/-s.d.) was -1.56 (+/-1.3). Patients were divided in two groups according to age at Tx: group I (n = 10), younger than 1.0 yr, and group II (n = 26) older than 1.0 yr. Median age (range) at Tx in group I was 0.8 yr (0.7-1.0) and in group II it was 3.35 yr (1.25-12.6). Thirteen patients (nine in group I) were receptors of living related donors. We evaluated linear growth, liver and renal function, immunosuppressive regimen and allograft rejection episodes. We did not find any significant differences in allograft or renal function, immunosuppressive therapy and number of acute rejection episodes or height Z score at Tx, second and third year post-Tx between both groups. The mean height Z score at Tx in group I was -1.61 and in group II -1.54; at the second year, group I -0.66 and group II -1.08; at the third year, group I -0.17 and group II -0.85; and at the seventh year (total group) -0.3. However, the height gain at the third year was better in group I than in group II (p < 0.01, t-test). Height Z score at the third year improved more than 1 SDS in seven out of eight patients in group I and in only nine out of 24 in group II (odds ratio 11.6). We also found a correlation between height gain at the third year and age at Tx (r-0.65) and between height gain at the third year and height Z score at Tx (r-0.54) (Pearson, p < 0.05). Children with BA who are transplanted before 12 months of age presented better catch-up growth without change survival and morbidity. Orthotopic liver Tx improves survival and also enables height gain in these children.     

APRI评价胆道闭锁和胆汁淤积综合征肝脏纤维化程度的临床意义

目的 探讨肝功能检查和门冬氨酸氨基转移酶/血小板指数(aspartate aminotransferase-to-platelet ratio index,APRI)与肝脏纤维化程度的关系,阐述其在BA肝纤维化评估中的临床价值.方法 收集2006年2月至2011年8月间在我院治疗的胆道闭锁患儿38例和胆汁淤积综合征患儿25例为研究对象.临床观察指标包括肝功能检查,肝脏活检切片,血小板指数;肝硬化程度采用Metavir分类,APRI的诊断性评估采用ROC曲线,应用SPSS 16.0统计学软件进行统计分析.并对本组患儿进行随访,随访时间是3～69个月(平均随访时间:20.7个月).结果 胆道闭锁组患儿ALP、γ-GT、DBIL(564.14±257.75、153.36±97.47、7.55±2.57)较胆汁淤积综合征组患儿存在明显升高(P＜0.05);胆道闭锁肝硬化组患儿Age、ALT、AST、γ-GT(84.50±24.72、225.07±109.68、331.64±130.93、951.07±667.24)较非肝硬化组明显升高,两组差异具有统计学意义(P＜0.05);胆汁淤积综合征肝纤维化组患儿Age、ALT、AST(84.76±14.28、159.92±61.76、238.15±62.60)较非肝纤维化组(54.17±11.17、98.92±58.08、151.17±41.44)明显升高,两组差异具有统计学意义(P＜0.05).患儿绘制APRI的ROC曲线,用于判定肝硬化程度,胆道闭锁组敏感性为79％,特异性为88％;胆汁淤积综合征组敏感性为91％,特异性为79％.胆道闭锁中肝硬化组病死率显著高于非肝硬化组,且自体肝生存情况低于非肝硬化组.结论 肝功能检查可以作为胆道闭锁的初步判断指标,绘制APRI的ROC曲线对于评价胆道闭锁及胆汁淤积综合征患儿的肝脏纤维化情况均有较高准确性和可靠性,可用于预测预后和提早做好肝移植准备,因其简单、无创性可以在临床上广泛应用.     

血清内毒素水平在胆道闭锁患儿术后的意义

目的探讨血清内毒素(ET)、透明质酸(HA)与胆道闭锁(BA)患儿术后肝损害、肝纤维化的关系。方法以行肝门空肠吻合(Kasai)术后的BA患儿为BA组(n=38),正常体检儿童(n=12)为对照组;BA患儿根据其胆红素水平再分成黄疸组(n=14)和无黄疸组(n=24)。由自动生化仪测肝功能,鲎三肽基质显色法测ET水平、放射免疫法测HA水平。结果 BA组患儿存在肝损害,血清ET、HA水平均较正常对照儿童高(P<0.05);BA组患儿中,黄疸患儿的肝损害较无黄疸者重,血清ET、HA水平也较无黄疸者高,差异均有统计学意义(P<0.05);血清ET水平与血清总胆红素、谷草转氨酶及HA水平呈正相关(P<0.05)。结论血清ET水平与术后BA患儿肝损害和肝纤维化密切相关,可作为术后判断BA患儿预后的有用指标。     

Time-course changes in the liver of biliary atresia patients on magnetic resonance imaging

Background:  Using magnetic resonance imaging (MRI), changes in the livers of postoperative biliary atresia (BA) patients were investigated.
Methods:  Periodic MRI was performed in 32 postoperative BA patients. The findings were evaluated by calculating the near-normal liver tissue area that corresponded with normal- or high-signal regions on T1-weighted imaging. The patients were divided into three groups based on the extent of near-normal liver tissue on the final MRI: group A, n  = 14; group B, n  = 13; and group C, n  = 5, included patients with >40%, 20–40%, and <20% area of near-normal liver tissue, respectively. The relationship among the macroscopic and histological findings in the liver at orthotopic living donor liver transplantation (OLDLT), patient outcomes, and MRI findings were investigated.
Results:  In group A, 11 patients had no evidence of liver dysfunction. In group B, six patients either had undergone or were awaiting OLDLT. In group C, all patients had undergone OLDLT. All patients had either adequate or impaired bile drainage in each liver segment. The segmental changes corresponded with the liver architecture at OLDLT. The changes could be evaluated on MRI at 1–2 years after surgery.
Conclusions:  Adequate and restricted areas of liver tissue with near-normal structure were indicative of good and poor prognoses, respectively. Shortly after portoenterostomy, these segmental changes occurred and/or developed in each liver segment and could be detected on MRI. It is emphasized that patients with >40% area of near-normal liver architecture at the initial stages did not require OLDLT, while those with <20% area did require OLDLT.     

��������˹�ؾ���Ѫ֢3��?

目的研究肝外胆道闭锁及巨细胞病毒性肝炎患儿肝组织中的多药耐药蛋白3(MDR3)的表达,探讨MDR3与胆汁淤积的关系。方法收集2000年1月至2009年10月广西医科大学第一附属医院儿童病例,分为肝外胆道闭锁(EHBA)组20例,CMV肝炎组12例,正常对照组10例,采用免疫组化法检测肝组织中MDR3的表达情况,通过图像分析技术进行定量分析。结果 MDR3在正常对照组、EHBA和CMV肝炎组患儿肝组织均有不同程度表达,以肝细胞膜表达最为明显,MDR3在肝组织的阳性表达随γ-GT的增高而增强,呈正相关(r=0.438,P=0.003),CMV肝炎组、EHBA组和正常对照组MDR3表达吸光度值分别为0.13±0.02、0.18±0.06和0.10±0.03,MDR3在EHBA肝组织中的表达强度明显高于CMV肝炎组和对照组(P<0.05)。结论 MDR3蛋白水平与胆汁淤积程度密切相关,EHBA肝组织MDR3的上调可能为机体的适应性反应,以利于增强磷脂的转运,减轻对胆管上皮的损伤。     

First liver transplantation for biliary atresia in children: The hidden effects of non‐centralization

The aim of our study was to determine the impact of initial orientation for medical and surgical care of children with BA on procedures and outcomes of the first LT. We retrospectively analyzed charts of children with BA who underwent first LT between 2006 and 2015. Patients were divided into two groups for comparison: a single‐center management group (from diagnosis to transplantation) and a secondarily referred group (children referred after failure of KP). We focused analysis on disease severity at transplantation, blood transfusion, and overall survival. One hundred and eighty‐five children were included. The median delay between pretransplant check‐up and transplantation was shorter in patients secondarily referred. A severe undernutrition was observed in 23.7% of children secondarily referred compared to 11.1% in children with a single‐center management (P = .024). At transplantation, INR and factor V level were higher in single‐center group patients (respectively, 67% vs 55%, P < .001 and 61% vs 49%, P = .002). The total of red blood cell and fresh frozen plasma administrated during procedure was two times higher in patients secondarily referred. Finally, patients with a single‐center management had a higher overall 12 months of survival rate (92.1% vs 83.1%, P = .033). In a country without low‐density population issues, the authors advocate an early referring to transplant center to further improving LT outcomes in children with BA.     

Early occurrence of hepatocellular carcinoma in biliary atresia treated by liver transplantation

A case of liver transplantation for HCC complicating BA in an eight-month old infant is reported. HCC in BA is extremely rare. Screening of AFP and ultrasonographic examination should be performed regularly in patients with secondary biliary cirrhosis for early detection of HCC.     

Hedgehog 通路在先天性胆道闭锁发病机制中研究进展

先天性胆道闭锁(congenital biliary atresia,CBA)是一种新生儿早期胆管疾病,导致肝内肝外胆管不同程度的闭锁并造成肝组织纤维化伴肝胆管不可逆性损伤。它是新生儿阻塞性黄疸和小儿肝移植最主要的原因。其发病机制目前尚未研究清楚,是遗传与环境因素共同作用导致的复杂疾病。Hedgehog信号通路在哺乳动物胚胎发育、器官形成及发育中有重要作用。近期研究发现,Hedgehog信号通路在CBA的发生过程中也发挥重要作用。Hedgehog通路的激活会影响胚胎肝脏发育、增强肝胆管纤维化的发生并且阻碍胆管形态的形成,从而导致CBA的发生。文章重点介绍Hedgehog通路在CBA形成过程中所起的重要作用。