胆道闭锁患者肝内胆管γδT细胞和调节性T细胞浸润及意义

目的研究胆道闭锁(Biliary atresia,BA)患者肝组织中γδT细胞和调节性T细胞(Foxp3+Treg)的比例变化。方法采用免疫组织化学方法和流式细胞术观察和检测胆道闭锁患儿组(BA组)23例和对照组(CG组)12例肝组织中γδT细胞分布情况以及γδT细胞和Foxp3~+Treg细胞比例关系。结果免疫组织化学染色显示BA组肝脏汇管区胆管周围有大量γδT细胞和一定程度的Foxp3~+Treg细胞浸润。流式细胞术显示胆道闭锁肝组织中γδT细胞与Foxp3~+Treg细胞比例明显高于对照组(P0.05),且γδTT细胞与Foxp3~+Treg细胞比例呈显著负相关(P0.05)。结论胆道闭锁患儿肝组织中γδT细胞增多,或抑制Foxp3~+Treg细胞增值,促进了胆管的进行性炎症损伤。     

胆道闭锁肝脏CX3CR1^(+)T淋巴细胞的比例与肝纤维化及预后的相关性研究北大核心CSCD

目的探讨表达CX3C趋化因子受体1(CX3C chemokine receptor 1,CX3CR1)的T淋巴细胞(即CX3CR1^(+)T淋巴细胞)在不同儿童肝脏疾病中的表达情况,以及胆道闭锁肝组织浸润的表达CX3CR1的T淋巴细胞亚群与胆道闭锁患儿临床指标以及预后之间的关系。方法回顾性分析2019年1月至2020年6月在广州市妇女儿童医疗中心接受手术的24例胆道闭锁患儿临床资料,同时选取同期进行手术的胆总管囊肿20例(接受胆总管囊肿根治手术)、胆汁淤积症4例(术中胆道造影排除BA)、门静脉高压9例(行门静脉高压治疗手术)作为对照组。对24例胆道闭锁患儿肝脏标本进行肝纤维化评分。利用流式细胞技术对所有患儿肝脏中浸润的T淋巴细胞亚群进行分析。比较T淋巴细胞亚群在不同疾病患儿肝脏中的表达差异。采用Spearman检验对BA患儿肝脏中CX3CR1^(+)CD8^(+)T淋巴细胞比例与肝功能指标、BA肝纤维化评分分别进行相关性分析。对BA患儿随访2年,比较预后良好及预后不良患儿肝脏表达CX3CR1的T淋巴细胞比例。结果胆道闭锁组肝脏CD69-CX3CR1^(+)CD8^(+)T淋巴细胞比例为(7.91±8.81)%,低于胆总管囊肿患儿(21.53±13.85)%以及门静脉高压患儿(13.82±7.75)%,差异具有统计学意义(P<0.001,P=0.016);胆道闭锁组肝脏CD69-CX3CR1^(+)CD4^(+)T淋巴细胞比例为(3.96±5.39)%,低于胆总管囊肿患儿(8.76±9.16)%以及门静脉高压患儿(7.68±5.98)%,差异具有统计学意义(P=0.020,P=0.003)。胆道闭锁肝脏CX3CR1^(+)CD8^(+)T淋巴细胞比例与患儿γ-谷氨酰转肽酶水平及直接胆红素水平呈负相关,差异具有统计学意义(P=0.002和P=0.048)。胆道闭锁肝纤维化评分与肝脏中CX3CR1^(+)CD8^(+)T淋巴细胞比例及CX3CR1^(+)Ki67^(+)CD8^(+)T淋巴细胞比例呈负相关(P=0.025,P=0.030)。预后良好患儿肝脏中CX3CR1^(+)T淋巴细胞的比例较预后不良患儿高(P<0.05)。结论CX3CR1^(+)CD4^(+)T淋巴细胞和CX3CR1^(+)CD8^(+)T淋巴细胞可以反映肝脏的免疫状态,参与肝纤维化,可能是预测胆道闭锁患儿预后的重要指标。     

紫癜性肾炎患儿CD4+CD25+调节性T细胞和淋巴细胞亚群变化的意义

目的 探讨紫癜性肾炎(HSPN)患儿外周血CD4+CD25+调节性T细胞百分比和淋巴细胞亚群的变化和意义.方法 初诊初治的HSPN患儿35例于治疗前抽取清晨空腹静脉血2 mL,加入乙二胺四乙酸(EDTA)钾盐抗凝,采用流式细胞仪(FCM)检测其外周血CD4+CD25+调节性T细胞百分比及淋巴细胞亚群数量,并分析CD4+CD25+调节性T细胞与24 h尿微量清蛋白(24 h UMA)的关系.同时随机选取同期本院35例健康体检儿童外周血标本作为对照组.结果 HSPN患儿外周血CD4+CD25+T细胞及CD4+CD25+/CD4+均低于健康对照组(Pa＜0.01);CD3+T细胞、CD4+T细胞、CD4+/CD8+、CD3-CD(16+56)+均低于健康对照组(Pa＜0.05);而CD19+CD23+则高于健康对照组(P＜0.05).CD4+CD25+调节性T细胞百分比与24 h UMA呈显著负相关(r=-0.61 P＜0.01). 结论 外周血淋巴细胞亚群的变化可能参与HSPN的发病过程,而CD4+CD25+调节性T细胞百分比可作为监测HSPN患儿病情及预后治疗的指标之一.     

102例胆道闭锁Kasai术后胆管炎分析

目的分析102例胆道闭锁Kasai术后胆管炎的发生情况,探讨其与预后的关系,以加强对胆道闭锁Kasai术后胆管炎的诊治。方法对2009年1月至2016年6月山西省儿童医院行Kasai手术并统一术后治疗方案的102例胆道闭锁患儿进行随访,随访内容包括Kasai术后胆管炎发作的时间、次数及频率,术后是否遵嘱服药、治疗情况及疗效等,分析胆管炎与自体肝存活率、黄疸消退率及肝功能恢复等预后情况的关系。结果术后未发生胆管炎组及胆管炎发作组2年累计自体肝存活率分别为65.5%和41.1%(P=0.030)。早期胆管炎发作组和晚期胆管炎发作组其2年累计自体肝存活率分别为22.2%和52.2%(P=0.013)。早期胆管炎发作组和晚期胆管炎发作组的黄疸消退率分别为33.3%和67.4%(P=0.007)。早期胆管炎发作组和晚期胆管炎发作组的肝功恢复良好率为44.4%和73.9%(P=0.023)。频发胆管炎组与偶发胆管炎组的2年自体肝生存率分别为19.4%和57.1%(P=0.002),频发胆管炎组和偶发胆管炎组的黄疸消退率分别为38.7%和66.7%(P=0.001),频发胆管炎组和偶发胆管炎组的肝功能恢复良好率分别为45.2%和76.2%(P=0.008)。结论胆管炎尤其早期、频发胆管炎影响自体肝存活率、黄疸消退率及肝功能恢复,最终影响胆道闭锁预后。提高医患双方对胆道闭锁术后胆管炎的认识,加强对胆管炎的防治,对提高胆道闭锁生存率有重要意义。     

CD4+CD25+Foxp3+调节性T细胞与IL-33在儿童哮喘发病机制中的作用

目的 探讨CD4+CD25+Foxp3+调节性T细胞(Treg)与IL-33在儿童哮喘发病中的作用.方法 采用流式细胞仪检测45例哮喘患儿(哮喘组)、50例呼吸道合胞病毒感染喘息患儿(喘息组)及40例健康儿童(对照组)外周血CD4+CD25+Foxp3+Treg细胞百分比,采用ELISA法检测各组外周血血清IFN-γ、IL-4、IL-5及IL-33浓度,进行比较分析.结果 哮喘组患儿体内CD4+CD25+Foxp3+Treg 水平较喘息组及对照组均降低(P<0.05);哮喘组患儿体内IL-33水平较喘息组及对照组均升高(P<0.05),哮喘组患儿体内CD4+CD25+Foxp3+Treg与IL-33呈负相关(r=-0.156,P<0.01).结论 在哮喘患儿发病机制中,CD4+CD25+Foxp3+Treg与IL-33可能存在相互作用.     

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.     

胆道闭锁肝脏CX3CR1+T淋巴细胞的比例与肝纤维化及预后的相关性研究

目的探讨表达CX3C趋化因子受体1(CX3C chemokine receptor 1, CX3CR1)的T淋巴细胞(即CX3CR1+T淋巴细胞)在不同儿童肝脏疾病中的表达情况, 以及胆道闭锁肝组织浸润的表达CX3CR1的T淋巴细胞亚群与胆道闭锁患儿临床指标以及预后之间的关系。方法回顾性分析2019年1月至2020年6月在广州市妇女儿童医疗中心接受手术的24例胆道闭锁患儿临床资料, 同时选取同期进行手术的胆总管囊肿20例(接受胆总管囊肿根治手术)、胆汁淤积症4例(术中胆道造影排除BA)、门静脉高压9例(行门静脉高压治疗手术)作为对照组。对24例胆道闭锁患儿肝脏标本进行肝纤维化评分。利用流式细胞技术对所有患儿肝脏中浸润的T淋巴细胞亚群进行分析。比较T淋巴细胞亚群在不同疾病患儿肝脏中的表达差异。采用Spearman检验对BA患儿肝脏中CX3CR1+CD8+T淋巴细胞比例与肝功能指标、BA肝纤维化评分分别进行相关性分析。对BA患儿随访2年, 比较预后良好及预后不良患儿肝脏表达CX3CR1的T淋巴细胞比例。结果胆道闭锁组肝脏CD69-CX3CR1+CD8+T淋巴细胞比例为(7.91±8.81)...     

肾母细胞瘤患儿外周血调节性T细胞和自然杀伤T细胞的表达研究

目的 研究肾母细胞瘤患儿外周血CD4+CD25+CD127low 调节性T细胞（Treg）和CD3+CD16+CD56+自然杀伤T细胞（NKT）的表达变化,初步探讨发生改变的原因及其临床意义。方法 选取2015年11月至2016年7月就诊的21例肾母细胞瘤患儿作为病例组,21例于本院体检的同年龄段健康儿童为健康对照组,采用流式细胞术检测两组儿童外周血中 CD4+CD25+CD127low T细胞占CD4+T细胞百分比和CD3+CD16+CD56+T细胞占CD3+T细胞百分比,分别代表Treg水平和NKT水平。结果 病例组肾母细胞瘤患儿外周血Treg水平低于健康对照组（P < 0.05）;病例组肾母细胞瘤患儿外周血NKT水平高于健康对照组（P < 0.05）。结论 Treg和NKT水平变化与肾母细胞瘤的发生与发展有关,Treg和NKT水平可能是反映肾母细胞瘤患儿免疫功能状态的较好指标。     

哮喘小鼠气道重塑过程中CD4~+CD25~+调节性T细胞和Th17细胞表达的动态变化

目的探讨哮喘小鼠Th17细胞和CD4+CD25+调节性T细胞(Treg)在脾组织中表达水平的变化规律及与气道重塑的关系。方法将SPF级雌性Balb/c小鼠48只随机分为对照组和哮喘组,利用腹腔注射卵清蛋白(OVA)和氢氧化铝混悬液致敏及雾化吸入OVA激发制备哮喘气道重塑模型,对照组应用生理盐水替代。两组分别在雾化2周、4周及8周后的24 h内随机处死8只小鼠,左肺组织病理切片观察肺气道重塑程度;流式细胞仪测定脾组织中Th17、CD4+CD25+Treg细胞占CD4+T细胞的百分比。结果随着激发时间的延长,哮喘组支气管总管壁面积(Wat/Pbm)及支气管平滑肌面积(Wam/Pbm)逐渐增厚(P0.01);脾组织细胞悬液中Th17细胞水平逐渐升高,与气道重塑的程度呈正相关(P0.01),而CD4+CD25+Treg细胞水平逐渐降低,与气道重塑程度均呈负相关(P0.01)。结论哮喘小鼠气道重塑是动态变化的过程,激发的时间越长,气道重塑越严重,Th17细胞表达越高,CD4+CD25+Treg细胞表达越低,两者间免疫失衡可能是哮喘气道重塑发生的重要因素之一。     

胆道闭锁汇管区炎性细胞浸润及其细胞因子的表达研究

目的 了解胆道闭锁(biliary atresia,BA)肝内汇管区炎症反应类型及细胞因子表达和具体作用机制.方法 应用免疫组织化学、免疫荧光和免疫印迹的方法对18例胆道闭锁患儿与15例先天性胆管扩张症(congenital biliary dilatation ,CBD)患儿肝脏标本进行对比研究.结果 BA患儿肝内汇管区大量CD4+、CD8+、CD68+细胞浸润(与CBD组相比均P＜0.01).肝内Th1通路相关细胞因子IFN-γ(干扰素-γ)、TNF-α(肿瘤坏死因子-α)表达,而CBD组为阴性.Th2相关细胞因子IL-4(白介素-4)、IL-5(白介素-5)及CD8+相关细胞因子粒酶、穿孔素表达在BA组与CBD组均为阴性.BA组肝内iNOS(诱导型一氧化氮合酶)检测阳性,胆管上皮Fas(自杀相关因子)表达(0.60±0.13)与CBD组(0.29±0.16)相比明显上调(P＜0.01),上皮细胞凋亡指数(49±13)%远大于CBD组(22±16)%(P＜0.01).结论 BA肝内汇管区炎症反应强烈,是由CD4+细胞Th1亚型及其相关细胞因子所介导的原发特异性炎症.巨噬细胞参与的Th1炎症通路可能在BA胆管上皮进行性损伤中起着关键作用.     

原发性肾病综合征患儿外周血CD4+CD25+调节性T细胞及CD19+CD23+细胞水平的变化及其意义

目的通过观察原发性肾病综合征(PNS)患儿外周血淋巴细胞亚群,尤其是CD4+CD25+调节性T细胞及CD19+CD23+细胞水平的变化,探讨其免疫发病机制。方法采用双标法用流式细胞仪检测25例初发PNS患儿(PNS组)外周血T淋巴细胞亚群(CD3+、CD3+CD4+、CD3+CD8+、CD4+CD25+)、B淋巴细胞亚群(CD3-CD19+、CD19+CD23+)及自然杀伤细胞(CD3-CD16+56+)水平,同时选取同期19例健康儿童作为健康对照组。数据采用SPSS 15.0软件进行统计学分析。结果 PNS组患儿外周血中CD3+、CD3+CD8+、CD4+CD25+、CD19+CD23+淋巴细胞均显著高于健康对照组(Pa<0.05),而自然杀伤细胞则较健康对照组显著降低(P<0.05);PNS组CD3+CD4+、CD4+/CD8+、CD3-CD19+淋巴细胞与健康对照组比较差异无统计学意义。结论体内淋巴细胞亚群的紊乱参与了PNS的发病过程,其中CD4+CD25+调节性T细胞及CD19+CD23+细胞的变化为PNS的免疫治疗目标提供了理论依据。     

CD8＋ T细胞在肠道病毒71型感染重症手足口病中的作用研究

目的：检测分析肠道病毒71型（EV71）感染手足口病患儿外周血CD8＋T细胞数量变化与患儿年龄以及病情严重程度之间的关系,进而分析探讨CD8＋T细胞在EV71感染神经系统并发症发生中的潜在作用。方法收集2014年3月至9月昆明市儿童医院感染科确诊的手足口病患儿138例,其中普通型33例,重型45例,危重型60例。患儿年龄9个月~5岁。采用流式细胞术对外周血中CD8＋T细胞亚群进行检测。结果与各年龄段健康儿童CD8＋T细胞参考值相比,除~2岁年龄段普通型手足口病患儿CD8＋T细胞百分比增高明显,~5岁年龄段危重型手足口病患儿CD8＋T细胞减低外,其他各年龄段各病情患儿中CD8＋T细胞均增高或略高。其中,9~15个月普通型、重型患儿外周血CD8＋T细胞百分比增高均较明显,而危重型患儿略增高;~2岁患儿随病情加重CD8＋T细胞增高幅度逐渐减低;~5岁年龄段患儿,普通型增高不明显,重型略高,危重型减低。~2岁年龄段普通型与重型、危重型手足口病患儿CD8＋T细胞百分比均存在显著差异（ P均﹤0.05）;而其他年龄段手足口病患儿,不同病情严重程度者CD8＋T细胞的表达无显著差异（ P均﹥0.05）。结论 CD8＋T细胞的表达变化与患儿年龄及病情严重程度间存在一定的关系;尤其是~2岁年龄段手足口病患儿体内CD8＋T细胞的表达减低与患儿病情发展至重症相关,推测CD8＋T细胞在~2岁年龄段手足口病患儿很可能发挥重要的抗病毒免疫反应。     

胆道闭锁患儿外周血T细胞ITGAL基因启动子区甲基化状态及其mRNA表达

目的 研究胆道闭锁（BA）患儿外周血T细胞ITGAL基因启动子区DNA甲基化状态及其对mRNA表达的影响。方法 选取2010年4~8月于复旦大学附属儿科医院（我院）初诊、并经外科手术病理学检查证实为BA的患儿为研究对象,分为BA组和甲基化结果验证组;选取我院同期行斜疝手术、日龄≤120 d和肝功能、肾功能正常的患儿为对照组。分离BA组和对照组CD4+和CD8+T细胞,提取DNA和RNA,行ITGAL基因启动子区DNA甲基化水平和mRNA表达水平检测。甲基化结果验证组分离细胞后,予5-氮杂胞苷干预和培养后,行甲基化水平和mRNA表达水平检测,验证研究结果。结果 BA组和对照组各20例进入研究,两组年龄和性别均匹配。①BA组和对照组CD4+和CD8+T细胞ITGAL基因启动子序列-250~250 bp均未发生甲基化。BA组CD4+T细胞-1450~-950 bp的CG二核苷酸平均甲基化水平显著高于CD8+T细胞（0.94 vs 0.75,P=0.02）,也显著高于对照组CD4+T细胞（0.94 vs 0.66,P<0.001）。②BA组外周血CD4+T细胞ITGAL mRNA表达显著低于CD8+T细胞（0.021±0.002 vs 0.032±0.004,P=0.013）,也显著低于对照组（0.021±0.002 vs 0.031±0.003,P=0.007）。BA组CD8+T细胞ITGAL mRNA表达与对照组差异无统计学意义（0.032±0.004 vs 0.034±0.006,P=0.266）。③甲基化验证组纳入5例BA患儿。验证结果显示,5-氮杂胞苷干预后CD4+和CD8+ T细胞ITGAL启动子区平均甲基化水平均显著低于未予5-氮杂胞苷干预的水平;ITGAL mRNA的表达均显著高于未予5-氮杂胞苷干预的水平。结论 BA患儿外周血CD4+ T细胞ITGAL启动子区发生高甲基化,并对mRNA表达产生影响。     

Analysis of changes in the percentage of B (CD19) and T (CD3) lymphocytes,subsets CD4, CD8 and their memory (CD45RO), and naive (CD45RA) T cells in children with immune and non-immune thyroid diseases

Graves' disease (GD) is an autoimmune thyroid disease caused by immunological abnormality. The immune cells (lymphocytes T and B) which infiltrate the thyroid gland play a key role in the development of autoimmune thyroid disease (AITD). The aim of this study was to evaluate the differences between distribution of T (CD3) lymphocytes, subsets CD4, CD8, and their memory (CD45RO), and naive (CD45RA) T cells and B (CD19) lymphocytes in the peripheral blood of patients with Graves' disease (GD) (n = 33, mean age 15.9 +/- 5.9 years) and non-toxic nodular goiter (NTNG) (n = 25, mean age 15.2 years), in comparison to age- and sexmatched healthy control subjects (n = 25, mean age 15.9 years). The percentages of peripheral blood lymphocyte subsets were analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. In the untreated Graves' patients we observed an increase in the percentage of CD19+ (p<0.007, p<0.003), CD4+ (p<0.004, p<0.017), CD4+CD45RO+ (p<0.04, NS), CD4/CD8 ratio (p<0.002, p<0.001) and a decrease in the percentage of CD8+ (p<0.02, p<0.02), CD4+CD45RA+ (p<0.04, p<0.03) cells in comparison to the healthy control subjects and euthyroid Graves' patients. These abnormalities were absent in children with non-toxic nodular goiter. In addition, the levels of CD3+, CD4+CD8+, CD8+CD45RO+ T cells and CD8 lymphocytes co-expressing CD45RA and CD45RO antigens were similar in all groups and no statistically significant differences were found in comparison to the healthy controls. In the untreated Graves' patients we found a positive correlation between serum levels of fT4 and fT3 and the percentage of CD19+ lymphocytes (r = 0.45, p<0.01, r = 0.37, p<0.04), between serum level of fT4 and the percentage of CD4CD45RO (r = 0.4, p<0.02) lymphocytes and between concentration of TRAb and CD4+ (r = 0.38, p <0.04) and CD19+ (r = 0.39, p<0.016) cells. Statistically significant negative correlations existed between TRAb, TPO-Ab or TG-Ab concentration in blood serum and the percentage of CD8+ lymphocytes (r = -0.55, p<0.002; r = -0.41, p<0.02; r = -0.51, p<0.004), and between fT4 concentration and the percentage of CD8+ (r = -0.39, p<0.02) lymphocytes. No such correlation was detected in patients with non-toxic nodular goiter. We conclude that the abnormal distribution of B lymphocytes, memory and naive T cell subsets in the peripheral blood in children and adolescents with untreated Graves' disease suggests their role in the development of autoimmunity. The normalization in the percentage of these immune cells after thyrostatic treatment in comparison to newly diagnosed patients confirms the immunomodulatory effect of methimazole therapy.     

儿童重症化脓性脑膜炎CD3＋ CD8＋ T细胞与炎症指标、体液免疫的变化

目的：通过观察儿童重症化脓性脑膜炎早期血CD3＋CD8＋T细胞的变化,以及与炎症指标、体液免疫指标之间的关系,探讨其在儿童重症化脓性脑膜炎发生发展中的临床意义。方法回顾性分析中国医科大学附属盛京医院PICU 2014年8月1日至2015年12月31日收治的39例1个月~14岁的重症化脓性脑膜炎患儿,血CD3＋CD8＋T细胞计数正常或升高(≥190个/mm3)为A组( n＝22),降低(<190个/mm3)为B组(n＝17),分析患儿的一般资料、血液炎症指标、体液免疫、脑脊液改变在两组患儿中的分布和差异。结果17例(43.6％)患儿CD3＋CD8＋T细胞明显下降;所有4例死亡均为B组患儿;虽然没有统计学差异,但 B 组 Glasgow 昏迷评分<8分者比例(58.8％)高于 A 组(31.8％)。B组C-反应蛋白、降钙素原中位数(最小值-最大值)分别为251.0(26.2-417.0)mg/L、32.7(0.9-100.0)ng/L,远远高于A组的106.5(12.0-458.0)mg/L、4.5(0.1-200.0)ng/L,差异有统计学意义(P<0.05);B组中6例(35.3％)外周血WBC<4×109/L,而 A组为1例(4.6％),中性粒细胞>80％的比例A组为7例(31.8％),而B组为12例(70.6％),两组比较差异有统计学意义(P<0.05)。 B组14例(82.3％)患儿脑脊液中糖含量<2.0 mmol/L,高于A组[11例(50.0％)],两组比较差异有统计学意义(P<0.05)。结论儿童重症化脓性脑膜炎CD3＋CD8＋T细胞可能受到抑制,其与患儿脑功能损伤程度、炎症反应以及预后相关。可能对指导临床免疫制剂的应用有一定帮助。     

高效抗逆转录病毒治疗对艾滋病患儿CD8+T细胞活化分子CD38和人类白细胞抗原DR表达水平的影响及其与病毒载量的关系

目的 观察高效抗逆转录病毒治疗(HAART)对我国艾滋病患儿CD8+T细胞活化标志分子CD38和人类白细胞抗原DR(HLA-DR)表达水平的影响及其与病毒载量的关系.方法 对194例接受HAART的艾滋病患儿进行横断面研究,用流式细胞术检测CD4+、CD8+T细胞数,以及CD8+/CD38+和CD8+/HLA-DR+T细胞比例,RT-PCR检测血浆HIV RNA载量.并检测52名健康儿童CD8+/CD38+和CD8+/HLA-DR+T细胞比例作为正常对照.结果 本组中,135例病毒载量＜400 copies/ml,59例病毒载量≥400 copies/ml.病毒载量≥400 copies/ml组CD8+/CD38+T和CD8+/HLA-DR+T细胞水平显著高于病毒载量＜400 copies/ml组,差异有统计学意义(29.6±10.1 vs19.9±9.8;17.7±6.4 vs 9.6±6.1,P＜0.05);病毒载量＜400 copies/ml组,CD8+/CD38+T细胞接近正常水平,而CD8+/HLA-DR+T细胞仍高于正常水平,差异有统计学意义(19.9±9.8 vs 15.6±9.0;9.6±6.1 vs 5.8±3.3,P＜0.05).CD8+/CD38+和CD8+/HLA-DR+T细胞百分比均与病毒载量成正相关(前者相关系数R=0.403,P=0.03,后者相关系数R=0.569,P=0.09).结论 艾滋病患儿CD8+T细胞活化程度与病毒载量成正比,有效的HAART治疗能够显著地降低HIV感染者免疫活化程度,CD8+/CD38+和CD8+/HLA-DR+T细胞百分比可能是资源有限地区替代病毒载量检测的潜在指标.

Abstract：

Objective To study the expression of CD38 and HLA-DR on CD8 + T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART) and the relationship of immune activation and disease progression. Methods A cross-section study of 194 pediatric AIDS patients receiving HAART was carried out and 52 age-matched healthy children were recruited as control. The percentage of CD4+ , CD8+ , CD8+/CD38 + and CD8+/HLA-DR+ T cells was tested using flow cytometry, and HIVRNA in plasma was detected by quantitative RT-PCR. Results One hundred and ninety-four pediatric AIDS patients were divided into two groups according to the viral load: 59 patients with VL≥400 copies/ml and 135 patients with VL＜400 copies/ml. The percentage of CD8 +/CD38+ and CD8 +/HLA-DR+ T cells of patients with VL≥400 copies/ml was significantly higher than that of patients with VL ＜ 400 copies/ml (P ＜ 0. 05 ). Of patients with VL ＜ 400 copies/ml, the percentage of CD8 +/CD38 + T cells was nearly normal, and the percentage of CD8 +/HLA-DR+ T cells was higher than normal level ( P ＜ 0. 05 ). There was a positive correlation between percentage of CD8+/CD38+ and of CD8 +/HLA-DR+ T cells and viral load ( R = 0. 403, P = 0. 03 for the former and R = 0. 569, P = 0. 09 for the later). Conclusions Effective HAART could decrease immune activation of HIV-infected children significantly. And there was a positive correlation between percentage of CD8 +/CD38 + and of CD8 +/HLA-DR + T cells and viral load, suggesting that the two indicators might be used as the substitution of viral load in resource-limited areas.     

胆道闭锁患儿Kasai术后早期胆管炎相关危险因素分析

目的 探讨影响胆道闭锁患儿(biliary atresia,BA)经典Kasai术后早期胆管炎发生的相关因素.方法 对本组中35例BA患儿的临床资料应用二项分类logistic同归分析方法作回顾性分析.结果 胆道闭锁患儿术后近期胆管炎发生与术后胆汁引流效果、术后辅以激素治疗及术中预留胆支长度有显著相关性,而与患儿性别、手术日龄、术前总胆红素及肝功能、手术前后辅以熊去氧胆酸和苯巴比妥利胆退黄治疗、术中设置防反流瓣及术后抗感染力度等因素无关.结论 胆道闭锁患儿Kasai术后胆汁引流效果好,辅以激素治疗,同时术中预留胆支长度充分的BA患儿,其术后不易发生早期胆管炎;反之,患儿发生早期胆管炎风险增加.

Abstract：

Objective To evaluate the factors affecting the early post-operative cholangeitis after classical Kasai operation for biliary atresia (BA) in children. Methods The clinical data of thirty-five BA children from pediatric surgery department of Nanjing Childrens Hospital was retrospectively analyzed using binary logistic regression analysis. Results The occurrence of early cholangitis after classical Kasai operation was found to correlate with post-op biliary drainage effect, hormone therapy and the length of Roux-loop in operation. No correlation could be established between early cholangeitis and sex,age, total bilirubin and pre-op liver function, ursodeoxycholic and luminal therapy, the installation of anti-refluxing valve in operation and the dosage of antibiotics. Conclusions Good biliary drainage,hormone therapy and sufficient length of Roux-loop were associated with decreased risk of early cholangitis.     

Surgical modifications,additions, and alternatives to Kasai hepato-portoenterostomy to improve the outcome in biliary atresia

Kasai hepato-portoenterostomy (HPE) is the most widely used surgical technique to restore bile flow in biliary atresia (BA). We aimed to review literature on HPE substitutes and additions to Kasai especially in advanced BA (ABA). A PubMed search was done for surgical procedures apart from or along with Kasai HPE for BA. Additional procedures to prevent cholangitis were also reviewed. Procedures and outcome were analysed. Alternative procedures done by the authors have also been described briefly. Results have been compiled in this review article. In ABA, with portal hypertension and liver cirrhosis, Kasai HPE is associated with poor outcome, increased morbidity, and even mortality. Most require liver transplant (LT). Some alternatives to HPE include exploration at porta hepatis to assess the bile flow yet avoid the major surgery (HPE) as a bridge to LT. Conduit diversion may help to combat cholangitis resistant to steroid therapy. Stoma formation is not preferred in cases listed for LT due to high risk of bleeding. Hepatocyte infusion, stem cell therapy, and synthetic liver are the future options to meet the challenges in BA. Various alternative procedures may become handy in the future especially in ABA.     

Changes in epigenetic regulation of CD4+ T lymphocytesin biliary atresia

Biliary atresia (BA) is a virus-induced autoimmune disease associated with abnormal DNA methylation patterns that contribute to disease presentation. This study examined DNA methylation patterns, changes to genes associated with methylation regulation, and changes to the autoimmune-related gene interferon gamma (IFN-γ) in CD4+ T cells from BA patients. We demonstrated that genomic DNA isolated from CD4+ T cells harvested from infants presenting with BA were hypomethylated relative to healthy controls. In addition, DNA methyltransferase (DNMT1) and DNMT3a mRNA levels were significantly lower in BA CD4+ T cells compared with controls and methyl-DNA-binding domain proteins (MBD1) mRNA expression (but not MBD4 detected at higher levels in BA patients), which was significantly lower in CD4+ T cells from BA infants than in controls. DNMT1 expression positively correlated with global DNA methylation in BA CD4+ T cells. IFN-γ mRNA expression levels in BA patients were also significantly increased, and the IFN-γ gene promoter region was hypomethylated in BA CD4+ T cells compared with controls and negatively correlated with DNA methylation. These data suggest that methylation changes in CD4+ cells may contribute to BA disease presentation and progression by affecting the expression of genes associated with autoimmunity.     

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.