甘露糖结合凝集素基因54号密码子多态性与川崎病易感性的研究

目的　探讨甘露糖结合凝集素 (MBL)基因第 5 4号密码子多态性与中国汉族儿童川崎病的易感性和临床表型的关系。方法　应用聚合酶链反应 限制性内切酶片段长度多态性分析 ,对16 0例中国汉族健康儿童及 95例川崎病患儿的MBL基因多态性进行检测。结果　 ( 1)中国汉族健康儿童MBL基因多态性分布与文献报道的香港华人及高加索人种接近 ,无统计学差异。( 2 )川崎病患儿GAC等位基因频率高于健康对照组 ( 0 2 5 8vs 0 138,P <0 0 1) ,而GGC等位基因频率低于健康对照组 ( 0 74 2vs 0 86 2 ,P <0 0 1)。GAC型等位基因与中国汉族儿童川崎病密切相关 [OR =2 18,95 %CI(OR值的 95 %置信区间 )为 1 38～ 3 4 4 ,P <0 0 5 ]。 ( 3)在川崎病患儿中 ,GAC型等位基因携带者其病史有前驱感染史者多于GGC纯合子 (P <0 0 1)。结论　川崎病发病受遗传背景的影响 ,与MBL第 5 4号密码子基因多态性密切相关     

甘露糖结合蛋白基因多态性与川崎病相关性的研究

目的　探讨甘露糖结合蛋白 (MBP)基因第 5 4号密码子GGC→GAC变异与川崎病 (KD)的关系。方法　采用PCR RFLP法对中国汉族 16 0例健康儿童及 95例川崎病患儿的MBP基因多态性进行检测。结果　中国汉族健康儿童MBP基因多态性分布与文献报道的中国香港地区华人及丹麦高加索人群无统计学差异。川崎病患儿MBP基因GGC/GAC基因型频率明显高于健康对照组 (45 2 %vs 2 5 0 % ,P <0 0 5 ) ,而GGC/GGC基因型频率显著低于健康对照组 (5 1 6 %vs 73 8% ,P <0 0 5 )。川崎病患儿GAC等位基因频率明显高于健康对照组(2 5 8%vs 13 8% ,P <0 0 5 ) ,而GGC等位基因频率明显低于健康对照组 (74 2 %vs 86 2 % ,P <0 0 5 )。结论　川崎病发病受遗传背景影响 ,与MBP第 5 4号密码子基因多态性密切相关。     

过敏性紫癜P-选择素基因 -2123位点多态性检测与临床分析

目的：研究P-选择素基因 -2123位点多态性与儿童过敏性紫癜（HSP）发病的相关性。方法应用聚合酶链反应 限制性片段长度多态性(PCR-RFLP)分析方法对86例HSP患儿（其中40例合并肾炎）和70例健康对照进行基因多态性检测。结果与对照组比较,HSP患儿P-选择素基因 -2123位点GG基因型频率和G等位基因频率均明显增高（P＜0.05）。合并肾炎患儿与未合并肾炎患儿比较,该位点各基因型频率及等位基因频率差异均无统计学意义（P＞0.05）。结论 P-选择素基因 -2123位点多态性可能与儿童HSP发病有关。     

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目的研究T细胞免疫球蛋白域黏蛋白域蛋白-1(TIM-1)启动子-1454G/A基因多态性与儿童过敏性紫癜(HSP)的相关性,探讨HSP的遗传易感因素。方法2008年3月至2009年2月安徽医科大学第一附属医院、安徽省立儿童医院住院HSP患儿143例,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析、基因测序等技术测定HSP患儿和178名正常儿童(对照组)的TIM-1基因多态性,计算基因型和等位基因频率,比较各组间关系。结果HSP组基因型及等位基因频率与对照组比较差异无统计学意义(P>0.05),但63例伴肾脏损害者即紫癜性肾炎(HSPN)组与80例不伴肾脏受累组-1454G/A3个基因型间比较差异有统计学意义(P<0.05),且HSPN患儿携带-1454G等位基因频率增高(OR2.375,95%CI1.168～4.830,P<0.05)。结论TIM-1基因启动子-1454G/A基因多态性可能与HSP易感性无关联,但携带-1454G等位基因的患儿出现HSPN风险增高,提示TIM-1基因多态性可能在决定HSP患儿肾脏受累遗传易感性方面有重要作用。     

儿童过敏性紫癜性肾炎血管内皮生长因子基因多态性的研究

目的：探讨血管内皮生长因子(VEGF)-634G/C基因多态性与汉族儿童紫癜性肾炎(HSPN)的关系。方法：应用聚合酶链反应－限制性内切酶片段长度多态性（PCR－RFLP）技术对100例过敏性紫癜（HSP）汉族儿童进行VEGF -634G/C基因型分析,其中包括合并紫癜性肾炎50例（HSPN 组）,无合并肾炎者 50例（单纯HSP组）。50例年龄、性别匹配的健康汉族儿童作为对照组。并采用酶联免疫吸附试验(ELISA)检测各组血浆VEGF水平。结果：HSPN组VEGF-634CC基因型（32%）和C等位基因频率（56%）均高于对照组（分别为10%和33%）及单纯HSP组（分别为10%和35%,P<0.01）。HSP患儿中,CC基因型者肾炎的发生率比GG基因型者明显增加, 差异有显著性意义（76% vs 31%,P<0.01）。CC基因型者血浆VEGF水平(180.5±40.7 pg/mL)较CG(145.2±48.3 pg/mL)及GG (101.5±26.5 pg/mL)基因型者显著上升, 差异有显著性意义（P<0.05）。结论：VEGF-634G/C基因多态性与儿童HSPN的发生有关。C等位基因可能是儿童HSPN的易感基因。［中国当代儿科杂志,2009,11（6）：417-421］     

儿童紫癜性肾炎肿瘤坏死因子α基因多态性分析

目的：探讨肿瘤坏死因子α（TNF-α）-308G/A基因多态性与儿童紫癜性肾炎（HSPN）的关系。方法应用聚合酶链反应（PCR）扩增产物直接测序方法对110例过敏性紫癜（HSP组）汉族儿童进行TNF-α-308G/A基因型分析,包括紫癜性肾炎52例（HSPN组）,无合并肾炎58例（单纯HSP组）。酶联免疫吸附法检测血浆TNF-α水平。90例健康汉族儿童作为对照组。结果HSP组TNF-α-308基因型分布频率与对照组比较差异无统计学意义（P>0.05）。HSPN组GA基因型（29%）、A等位基因频率（18%）均明显高于单纯HSP组（分别为10%和7%,P<0.05）,其中GA+AA 基因型患儿血浆TNF-α水平（7.1±2.3 pg/mL）较GG基因型患儿（5.7±1.5 pg/mL）明显升高（P<0.05）。结论TNF-α-308G/A 基因多态性与HSP儿童肾脏损害的发生有关,A等位基因可能是儿童HSPN的易感基因。     

巨噬细胞移动抑制因子-173G/C基因多态性与紫癜性肾炎的相关性

目的探讨江西地区汉族儿童巨噬细胞移动抑制因子(MIF)-173G/C基因多态性与过敏性紫癜(HSP)、紫癜性肾炎(HSPN)的相关性。方法HSP患儿131例[并肾炎80例(HSPN组),未并肾炎51例(单纯HSP组)]与健康对照儿童105例,均于晨起空腹采集外周静脉血2 mL,应用Promega人全血基因组DNA试剂盒提取其外周血基因组DNA;PCR法扩增目的DNA片段,内切酶(AluⅠ)酶切消化产物,采用限制性片段长度多态性(RFLP)方法分析MIF-173位点多态性,凝胶电泳图像系统判断基因型。采用频率计数法计算单纯HSP组、HSPN组和健康对照组各基因型及等位基因频率。采用SPSS11.5软件对各基因型及等位基因频率差异进行χ2检验,计算比值比(OR)及95%可信区间(95%CI),以估计各基因型对疾病发生的相对危险度。结果MIF-173G/C位点检测出3种基因型,分别为GG型、GC型、CC型。单纯HSP组和健康对照组仅检出GG和GC型,而HSPN组3种基因型均检出。HSPN组MIF-173基因突变型(37.5%)和C等位基因频率(20.0%)均高于健康对照组(分别为20.0%、10.0%)(χ2=6....     

血管紧张素转换酶基因多态性对过敏性紫癜的影响

目的 探讨过敏性紫癜(HSP)与血管紧张素转换酶(ACE)基因多态性间的关系。方法 选择70例HSP患儿,其中无肾脏损害32例,过敏性紫癜性肾炎(HSPN)38例。HSPN中单纯性血尿15例,蛋白尿23例。健康对照儿童100例。采用聚合酶链反应(PCR)检测ACE基因型。结果 1.HSP患儿与健康对照组ACE基因型分布比较无差异(P>0.05);2.非肾脏损害患儿32例与伴单纯性血尿HSPN患儿15例ACE基因型分布比较亦无差异(P>0.05);但与23例伴蛋白尿HSPN患儿ACE基因型分布比较,缺失型(DD),型者发生蛋白尿频率明显升高(P<0.05)。结论 HSPN蛋白尿发生与ACE基因多态性相关,DD型者发生蛋白尿机会明显增多。     

T细胞免疫球蛋白域黏蛋白域蛋白-1-1454位点基因多态性与儿童过敏性紫癜的关系研究

目的研究T细胞免疫球蛋白域黏蛋白域蛋白-1(TIM-1)启动子-1454G/A基因多态性与儿童过敏性紫癜(HSP)的相关性,探讨HSP的遗传易感因素。方法2008年3月至2009年2月安徽医科大学第一附属医院、安徽省立儿童医院住院HSP患儿143例,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析、基因测序等技术测定HSP患儿和178名正常儿童(对照组)的TIM-1基因多态性,计算基因型和等位基因频率,比较各组间关系。结果HSP组基因型及等位基因频率与对照组比较差异无统计学意义(P0.05),但63例伴肾脏损害者即紫癜性肾炎(HSPN)组与80例不伴肾脏受累组-1454G/A3个基因型间比较差异有统计学意义(P0.05),且HSPN患儿携带-1454G等位基因频率增高(OR2.375,95%CI1.168～4.830,P0.05)。结论TIM-1基因启动子-1454G/A基因多态性可能与HSP易感性无关联,但携带-1454G等位基因的患儿出现HSPN风险增高,提示TIM-1基因多态性可能在决定HSP患儿肾脏受累遗传易感性方面有重要作用。     

CTLA-4基因多态性与过敏性紫癜的相关性研究

目的探讨CTLA-4基因多态性与儿童过敏性紫癜(HSP)的相关性。方法选取60例HSP患儿为病例组,其中男33例,女27例;另选取30例健康儿童为对照组。按有无肾脏损害将HSP患儿分为紫癜性肾炎(HSPN)组(n=30)和Non-HSPN组(n=30)。采用PCR-RFLP法,对CTLA-4基因+49及-1722位点各基因型及等位基因频率进行分析。结果 +49位点AA、AG、GG基因型及等位基因频率在病例组和对照组之间、HSPN组与Non-HSPN组之间、不同性别HSP患儿间比较差异均无统计学意义(P0.05)。-1722位点TT、TC、CC基因型及等位基因频率在病例组和对照组之间、不同性别HSP患儿间比较差异均无统计学意义(P0.05);CC基因型及T、C等位基因频率在HSPN组与Non-HSPN组间比较差异有统计学意义(P0.05)。将+49位点与-1722位点组合:各组合基因型频率在病例组和对照组之间、不同性别HSP患儿间比较差异无统计学意义(P0.05);GG+CC基因型组合在HSPN组与Non-HSPN组间比较差异有统计学意义(P0.05)。结论 CTLA-4基因+49位点A/G基因多态性与HSP发病无关;-1722位点CC基因型及C等位基因,以及+49位点GG与-1722位点CC组合基因型可能为HSPN发病的危险因素。     

甘露糖结合凝集素基因多态性与幼年特发性关节炎的相关性

目的 探讨甘露糖结合凝集素（MBL）基因外显子1多态性与幼年特发性关节炎（JIA）易感性的关系。方法 采用聚合酶链反应．限制性片段长度多态性（PCR，RFLP）法分析93例JIA和48名正常健康儿童MBL基因外显子1第54和57位密码子的多态性。结果 1．JIA及健康儿童均未发现有57位密码子突变。2．JIA患儿第54位密码子野生型基因型频率为71．0％（66／93），杂合子型为25.8％（24／93），突变纯合子型为3．2％（3／93）；正常对照儿童野生型75．0％（36／48），杂合子型为25．0％（12／48），未发现突变纯合子基因型；各基因型与JIA组比较无差异（P〉0．05）。结论 MBL基因外显子1多态性与JIA无相关。     

Association between mannose binding lectin polymorphisms and predisposition to bacterial meningitis

The aim of this study was to examine the presence of any association between mannose binding lectin (MBL) gene variants and bacterial meningitis. Codon 54 (B allele) and codon 57 (C allele) polymorphisms in exon 1 of the MBL gene were investigated in 50 healthy controls and 31 patients diagnosed as purulent meningitis. Codon 57 polymorphism was not found in our patient and control groups. B allele frequency was significantly higher in the patient group (22%) compared to the control group (3%). AB genotype was determined in 39% and 6% of patient and healthy control groups, respectively, and the difference was statistically significant. AA genotype was determined in 61% of the patient group and in 94% of the control group, and it was statistically low in the patient group. These results suggest that codon 54 polymorphism in the MBL gene may play a role in susceptibility to bacterial meningitis in children.     

The effect of mannose-binding protein gene polymorphisms in recurrent respiratory system infections in children and lung tuberculosis

Mannose-binding lectin (MBL) is able to bind pathogens as an opsonin and plays an important role in the innate immunity. The aim of the present study was to determine the frequencies of the MBL gene variants in the Turkish population and to examine the presence of any association between MBL variants and development of tuberculosis (TB) in adults and recurrent respiratory tract infections in children. Two structural gene mutations in exon 1 of MBL gene (codon 54 and codon 57) were studied. The overall distribution of genotypes did not significantly differ between controls and TB patients/children with recurrent respiratory system infections. The frequency of allele B was calculated as 0.14, 0.09 and 0.06 for control, TB patients and children with recurrent respiratory system infections, respectively. It was found to be significantly lower in children with recurrent respiratory system infections than in controls (chi2: 4.68, d.f: 1, p: 0.030).     

Mannose-binding lectin polymorphisms and recurrent respiratory tract infection in Chinese children

In order to establish the reference value of mannose-binding lectin (MBL) serum level in children and to investigate the correlation between the polymorphisms of MBL2 gene and serum MBL level in healthy Chinese of Han ethnic group and in children of Chinese Han ethnic group with recurrent respiratory tract infections (RRTI), the concentration of oligomerized MBL was measured by enzyme-linked immunosorbent assay, and MBL2 gene polymorphisms were analyzed by restriction fragment length polymorphism of polymerase chain reaction and polymerase chain reaction-sequence specific primer. The median MBL levels in the 470 normal children were 2536 ng/ml, and the P_2.5–P_97.5 was 161–5,070 ng/ml. Our research showed that two promoter polymorphisms at −550, −221 of start codon and coding variants at codon 54 of MBL2 gene affected the protein level significantly and the most frequent genotype in Hans is HYPA/HYPA. Our results also showed that serum MBL level was significantly lower in recurrent respiratory tract infections patients compared with healthy controls (Z, −3.04, P = 0.002). The frequency of the promoter LXP haplotype and the B allele was significantly higher in RRTI patients than in controls (χ ² 4.05, P < 0.05; OR 1.63, 95%CI 1.01∼2.62; χ ² 4.27, P < 0.05; OR 1.94, 95%CI 1.02∼3.68). Conclusion: We have established that the reference value of serum MBL level in Chinese aged between 0 and 6 years (161–5,070 ng/ml), and we found that LXP and the B are risk factors for RRTI.     

Age-dependent association of human mannose-binding lectin mutations with susceptibility to invasive meningococcal disease in childhood

BACKGROUND: Mannose-binding lectin (MBL) is an important factor of the innate immune system, and MBL-initiated complement activation is an important early defense mechanism against various bacterial infections, including invasive meningococcal disease. METHODS: In a pediatric cohort (ages 2-215 months) with invasive meningococcal disease, we investigated the overall and age-stratified frequency of 3 MBL exon 1 variations (C154T, G161A, G170A), previously shown to result in markedly decreased MBL plasma concentrations, by allele specific fluorescent hybridization probe real-time PCR assays and direct sequencing. Healthy age-matched volunteers with the same ethnic background and no history of meningococcal disease served as a control group. RESULTS: The overall frequency of a MBL exon 1 variant genotype was significantly higher in patients than in controls (31.8% vs. 8.2%, P < 0.001). In the patient group with disease onset less than 24 months of age, the prevalence of MBL structural variant genotype was further increased (39.3%; P < 0.001) and most pronounced in children with disease onset less than 12 months of age (57.1%; P < 0.001) when compared with healthy controls. Analysis of clinical severity and outcome revealed no significant difference between patients with wild-type and mutant alleles. CONCLUSIONS: Our data suggest that MBL exon 1 structural variants are significantly associated with susceptibility to childhood meningococcal disease in an age-dependent manner.     

调节性T细胞在儿童过敏性紫癜发病机制中的作用初探

目的系统观察过敏性紫癜(HSP)急性期调节性T细胞(Tr)亚群及辅助性T细胞亚群(Th1/Th2)的变化,探讨HSP急性期免疫失衡的发病机制。方法流式细胞术检测20例HSP急性期患儿各种调节性T细胞亚群(CD4+CD25+Tr、Tr1、Th3等)和辅助性T细胞亚群(Th1、Th2)的改变,并采用逆转录聚合酶链反应(RT-PCR)和荧光定量聚合酶链反应(Real-tim e PCR)检测其外周血单个核细胞(PBMC)Foxp3 mRNA的表达。同期20例同龄健康儿童作为对照。结果HSP急性期CD3+CD8-INF-γ-IL-4+(Th2)细胞显著增高(P<0.05),Th1/Th2比值显著降低(P<0.05)。各调节性T细胞亚群CD4+CD2+5Tr、CD4+IL-4-IL-10+(Tr1)、CD4+TGF-β+(Th3)细胞与正常对照组比较均显著降低(P均<0.05)。HSP组PBMC Foxp3 mRNA的表达与正常对照组比较亦显著降低(0.22±0.05vs.66.32±9.25,P<0.001)。结论HSP急性期存在明显的免疫失衡,Th2优势明显;调节性T细胞CD4+CD2+5Tr、Tr1、Th3数量减少导致的免疫抑制效应不足可能是导致HSP免疫失衡的重要原因,而HSP患儿调节性T细胞减少与Foxp3表达降低有关。     

The CBLB gene and Graves' disease in children

The CBLB gene functions as a negative regulator of autoimmunity. Impairment of the Cbl-b signaling pathway may contribute to human autoimmune disease. dbSNP rs2305035 is a C/T polymorphism located in exon 10 of the CBLB gene. We report an association study of this polymorphism in children with Graves' disease. The patients were 158 unrelated children (125 girls) with Graves' disease, aged 9.8 +/- 3.3 years. The controls consisted of 237 adults without a history of autoimmune disease. The C allele and phenotype frequencies of patients and controls were 247 (78.2%) vs 356 (75.1%) (OR = 1.19, p >0.05) and 151 (95.6%) vs 221 (93.2%) (OR = 1.56, p >0.05), respectively. The allelic polymorphism in patients and controls with and without DRB1*09012 were also not significantly different. This study demonstrates that the C/T polymorphism in exon 10 of the CBLB gene is not associated with Graves' disease in children.