CD40基因多态性与川崎病及其冠状动脉损伤的相关性研究

目的 探讨CD40 基因两个SNP 位点rs4810485 和rs1535045 的多态性与我国中部地区儿童川崎病（KD）以及动脉损伤遗传易感性的关系。方法 采取病例对照研究方法,分别选取184 例KD 患儿和206例正常体检儿童作为研究对象。利用PCR-RFLP 的方法测定两个SNP 位点多态性分布。结果 KD 患儿SNP位点rs4810485 的基因型（GG、GC、CC）频率及等位基因频率与正常对照组相比差异均无统计学意义（均P>0.05）;SNP 位点rs1535045 基因型（TT、TC、CC）与对照相比差异有统计学意义（P=0.011）,T 等位基因为风险因子（OR=1.592,95%CI：1.182~2.144,P=0.004）。患者中两个SNP 位点多态性均不与冠状动脉损伤相关（均P>0.05）。结论 CD40 基因SNP 位点rs4810485 与KD 无相关性, rs1535045 与KD 的易感性相关。     

GDF5基因rs224331多态性与汉族青少年体格发育指标关联分析

目的 探讨中国汉族青少年GDF5基因rs224331位点多态性与身高、体重和BMI的相关性。方法 采集江苏省某技校1 790名17~22岁青少年的身高、体重指标并计算BMI。采用Taqman探针real-time PCR方法检测GDF5基因rs224331位点的单核苷酸多态性,并分析该基因位点与男女生身高、体重和BMI的相关性。结果 97.9%（1 754/1 790名）基因分型成功,男生859名,女生895名。rs224331位点频率最高的基因型为AA（51.7%）,其次为AC（39.6%）,CC最少（8.7%）。rs224331的基因型分布与男生、女生的身高无显著关联（P分别为0.728和0.723）;rs224331不同基因型间的男生体重和BMI差异均有统计学意义（P均<0.01）,男生AC基因型体重和BMI均显著高于AA基因型（P=0.002）,女生中未发现类似的相关性（P分别为0.713和0.921）。结论 GDF5基因rs224331位点多态性在本研究的汉族人群中与身高无明显关联,提示该多态性位点可能存在种族特异性。rs224331位点多态性与男生体重和BMI存在相关性。     

雌激素受体α、β基因单核苷酸多态性与尿道下裂相关性研究CSCD

目的 研究雌激素受体α基因（ESR1）、β基因（ESR2）单核苷酸多态性（SNP）与尿道下裂易感性之间的关系。方法 选择2013年11月至2014年5月我院收治的384例尿道下裂患儿作为研究组,同期非尿道下裂患儿411例作为对照组。选取目前报道较多的热点SNP位点结合SNP公共数据库,对2个基因共13个SNP位点应用多重单碱基延伸SNP分型技术作基因分型检测。对每个位点的两种等位基因进行卡方检验,对共显性、频率低的等位基因为显性,频率低的等位基因为隐性及叠加作用4种遗传模型进行Logistic回归分析,同时进行单倍体分析。了解ESR1、ESR2单核苷酸多态性与尿道下裂发病的相关性。结果 检验结果显示,研究组与对照组比较,在ESR2基因的SNP位点rs1256040差异有统计学意义（P＝0.019）,其余位点检测结果差异均无统计学意义（P〉0.05）。使用SNPStats软件对每个位点进行回归分析,发现ESR2基因SNP rs1256040在共显性、频率低的等位基因为显性及叠加作用的3种遗传模型中差异有统计学意义（P值分别为0.041、0.014和0.016）。单倍体分析结果显示,研究组和对照组在ESR2-Block2（rs10483774和rs2987983）差异有统计学意义（P＝0.043）;其余各单倍体型中,两组频率分布差异均无统计学意义（P〉0.05）。结论 ESR2基因单核苷酸多态性可能和中国北方人群尿道下裂易感性相关,ESR2基因rs1256040位点的基因型G/A、G/G可能是决定尿道下裂易感性的一个重因素。     

ZNF365基因多态性与儿童支气管哮喘的相关性

目的 探讨我国中部湖北地区儿童ZNF365基因的两个SNP位点(rs2393903和rs10995251)的多态性与支气管哮喘及其临床特点的关系。方法 采用病例-对照的研究方法,选取湖北地区221例支气管哮喘患儿和243例同时期体检正常的儿童作为研究对象,利用限制性片段长度多态性分析的方法检测两个SNP位点的多态性分布。结果 哮喘患儿中SNP rs2393903位点的3种基因型(GG、GA、AA)分布及等位基因频率与正常对照组相比差异均无统计学意义;而SNP rs10995251位点的基因型(CC、CT、TT)以及等位基因频率与正常对照组相比差异有统计学意义(P< 0.05),C等位基因为风险因子(OR=1.380)。在哮喘患儿中SNP rs10995251位点CC基因型患儿血清免疫球蛋白E(IgE)的水平高于TT基因型患儿(P< 0.05)。结论 ZNF365基因SNP rs10995251位点的多态性与我国中部湖北地区儿童支气管哮喘的易感性相关;且该位点的多态性可能影响患儿血清IgE水平。     

IL1R1基因多态性与儿童哮喘的相关性

目的 探讨我国中部地区儿童IL1R1 基因的两个SNP 位点(rs1558641 和rs949963)的多态性与哮喘易感性的相关性。方法 采用病例-对照的研究方法,选取来自于我国中部地区的208 例哮喘患儿(哮喘组)和223 例同时期体检正常的儿童(健康对照组)作为研究对象。利用限制性片段长度多态性分析(PCRRFLP)的方法检测IL1R1 基因两个SNP 位点(rs1558641 和rs949963)的多态性分布;酶联免疫吸附试验(ELISA)测定血清中IL1R1 的水平。结果 哮喘组患儿SNP 位点(rs1558641)的基因型及等位基因频率与健康对照组相比差异无统计学意义。而哮喘组患儿SNP 位点(rs949963)GG 基因型的比例显著高于健康对照组(P=0.031),且两组等位基因频率差异也有统计学意义(P=0.018)。哮喘组血清IL1R1 的水平明显高于健康对照组(P=0.011),且SNP 位点(rs949963)GG 基因型的患儿血清IL1R1 水平高于其他基因型(AA+AG)的患儿(P=0.028)。结论 IL1R1 基因SNP 位点(rs949963)的多态性与我国中部地区儿童哮喘的易感性相关,且该位点的多态性可能影响患儿血清中IL1R1 的表达水平。     

特发性矮小患儿胰岛素样生长因子受体基因单核苷酸多态性研究

目的：探讨胰岛素样生长因子-I受体（IGF-IR）基因单核苷酸多态性（SNP）与特发性矮小（ISS）的发病危险的相关性。方法：2008~2011年确诊为ISS患儿804例,正常对照组575例,用Snapshot法检测两组 IGF-IR基因相关位点SNP。结果：两组rs1976667基因型分布频率差异无统计学意义（P>0.05）;rs1976667 A等位基因分布频率较正常对照组高,差异有统计学意义（P<0.01)。结论：IGF-IR基因rs1976667位点等位基因A是ISS发病的危险因素。     

胰岛因子1基因2个SNP位点与儿童先天性心脏病的相关性研究

目的：探讨天津地区汉族儿童胰岛因子1（Islet1,ISL1）基因2个单核苷酸多态性（SNP）位点rs41268421、rs1017与先天性心脏病（CHD）的相关性。方法：应用聚合酶链反应（PCR）和基因测序技术对35例CHD患儿和30名除外CHD儿童的rs41268421、rs1017位点进行检测,分别比较两个SNP位点基因型频率和等位基因频率在两组的分布情况,并进行单体型分析。结果：SNP位点rs41268421存在GG、GT、TT 3种基因型,CHD组T等位基因频率及携带T等位基因的基因型（包括GT和TT）频率高于对照组（P<0.05）,携带T等位基因的儿童患CHD的危险是G等位基因的4.833倍;rs1017 位点存在AA、AT、TT 3种基因型,CHD组T等位基因频率及携带T等位基因的基因型（包括AT和TT）频率高于对照组（P<0.05）,携带T等位基因的儿童患CHD的危险是A等位基因的4.491倍;2个SNPs位点得出4种单体型,以TT型儿童发生CHD的危险性最高（OR=7.813）。结论：天津地区汉族儿童中ISL1基因单体型TT的出现很可能会增加CHD的患病风险。     

IL-23受体和IL-17单核苷酸多态性与汉族早产儿坏死性小肠结肠炎的关系

目的 探讨白细胞介素-23受体（IL-23R）基因rs10889677位点、IL-17A基因rs2275913位点和IL-17F基因rs763780位点单核苷酸多态性（SNP）与汉族早产儿坏死性小肠结肠炎（NEC）的关系。方法 前瞻性选取2017年1月至2019年1月新生儿重症监护病房收治的100例汉族NEC早产儿为研究对象,其中Ⅱ期63例,Ⅲ期37例;另选取与NEC患儿胎龄、性别匹配的100例早产儿作为对照。采用PCR法和Sanger测序法鉴定rs10889677、rs2275913、rs763780位点的SNP。采用非条件logistic回归分析基因多态性与NEC易感性和病情严重程度的关系。结果 rs10889677位点、rs2275913位点基因型和等位基因频率对NEC发病无影响（P > 0.05）;rs763780位点基因型对NEC发病无影响（P > 0.05）,但C等位基因携带者相对于T等位基因携带者的NEC发病风险为1.652倍（95% CI：1.052~2.695,P < 0.05）。TC+CC基因携带者相对于TT基因携带者的NEC发病风险为1.856倍（95% CI：1.045~3.201,P < 0.05）。TC+CC基因携带者相对于TT基因携带者的NEC Ⅲ期的发生风险为2.965倍（95% CI：1.052~6.330,P < 0.05）;C等位基因携带者相对于T等位基因携带者的NEC Ⅲ期的发生风险为2.363倍（95% CI：1.034~4.093,P < 0.05）。结论 IL-23R基因rs10889677位点和IL-17A基因rs2275913位点SNP与汉族早产儿的NEC易感性无关,IL-17F基因rs763780位点TC+CC基因型和C等位基因与NEC易感性和NEC病情严重程度有关。     

Toll样受体9基因多态性与儿童EB病毒感染关系的探讨

目的通过检测Toll样受体9(TLR9)基因rs187084、rs5743836、rs352139、rs352140 4个单核苷酸多态性(SNP)位点基因型分析其SNP与儿童EBV感染的相关性。方法收集2013年2月至2014年2月吉林大学第一医院无EBV感染及EBV感染后不同临床表现类型的患者临床资料及外周静脉血标本,其中EBV感染阴性者54例,传染性单核细胞增多症(IM)患者56例,轻微临床症状型EBV活动性感染患者69例。应用盐析法提取血标本DNA,PCR扩增包含所选SNP位点的目的 DNA片段,PCR产物直接测序,分析SNP位点基因型,比较不同组别间基因型及等位基因分布状况。结果 (1)在实验人群中,位点rs187084基因型有TT、TC、CC 3种,rs352139基因型有AA、AG、GG 3种,rs352140基因型有GG、GA、AA 3种,最小等位基因频率分别为39.7%,38.8%,41.9%。以上3位点在实验人群中具有多态性,且与相关数据库中公布的其在中国人群中分布状况相似。而位点rs5743836基因型有TT、TC 2种,TC基因型在所有样本中仅发现1例,表明此位点在实验人群中不存在多态性。(2)在研究组间,各位点基因型及等位基因频率组间分布差异无统计学意义(P均>0.05)。结论在纳入研究的179例人群中,TLR9基因位点rs5743836不存在多态性,rs187084、rs352139、rs3521403个位点存在多态性,且分布情况与相关数据库中类似,未发现此3位点的基因多态性与儿童EBV感染及疾病的严重程度有相关性。     

IL-19基因多态性与儿童乙型肝炎病毒易感性研究

目的 探讨白介素19（IL-19）基因的单核苷酸多态性（SNP）与儿童乙型肝炎病毒（HBV）易感性的关系。方法 采用病例对照研究,收集HBsAg阳性儿童136例（病例组）,HBsAg阴性的健康儿童297例（对照组）,应用PCR 聚合酶链反应和DNA测序法进行基因分型。结果 IL-19基因的rs1798位点的基因型在病例组和对照组人群中频率分布差异有统计学意义,病例组的CG 基因型的比例显著高于对照组（P < 0.05）;IL-19基因的rs2243191位点的基因型和等位基因在HBV感染高危儿童感染组和未感染组人群中频率分布差异有统计学意义,感染组TC、CC基因型以及C等位基因的比例显著高于未感染组（P < 0.05）。结论 IL-19 基因SNP 位点rs1798的基因多态性可能与儿童乙型肝炎易感性相关;rs2243191的基因多态性可能与HBV感染高危儿童突破感染易感性相关。     

Infant formulas supplemented with prebiotics: intestinal microbiota and immune responses

It is well known that the type of feeding influences the composition of the gut microflora after birth. Human milk favours the growth of a 'bifidus flora' which, according to several evidences, may activate the immune system and defend from pathogens. Breast milk oligosaccharides, which are involved in many functional effects both at local and systemic level, are thought to stimulate the growth of health promoting microbes, such as bifidobacteria, and may ultimately influence the immune system. In accordance with this current working hypothesis, dietary modulation of the gut microbiota to obtain a 'bifidus flora' also in bottle-fed infants may be a useful way to stimulate immunological functions and to harbour a biological barrier against pathogens. In several clinical trials prebiotic oligosaccharides have been used to mimic the beneficial effects of breast milk oligosaccharides. A mixture of oligosaccharides has shown its efficacy in stimulating the establishment of a 'bifidus flora', with stools closer to those found in breast-fed infants. Several experimental data also indicate that oligosaccharides might modulate the immune system and contribute to the improvement of the protective properties of infant formulas.     

Neutral oligosaccharides in colostrum in relation to maternal allergy and allergy development in children up to 18 months of age

Several recent studies have demonstrated a relationship between the composition of the gut microbiota in infancy and subsequent development of allergic disease. Human milk is the major food in infancy and may thus profoundly influence the composition of the gut flora. Oligosaccharides in breast milk survive the passage through the stomach and are utilized by the gut microbiota. As the relationship between breast feeding and childhood allergy is controversial we hypothesized that the composition of oligosaccharides in breast milk might explain the controversy. Nine of the most abundant neutral oligosaccharides in human milk were analysed in colostrum samples from allergic and non-allergic women and related to subsequent development of allergy in their children. The carbohydrate fraction of the colostrum was separated by gel permeation chromatography and neutral oligosaccharides, tri- to hexasaccharides were collected. Neutral oligosaccharides were analysed with high-performance liquid chromatography. There was a large variation in the concentration of neutral oligosaccharides in colostrum, which could not be explained by the allergic status of the women. Allergic children consumed higher amounts of neutral oligosaccharides in total, although not significantly (p = 0.12). When different oligosaccharides were analysed separately, there was no significant difference in consumption between the infants who developed atopic allergy later (n = 9) and infants who did not (n = 11). Thus, the amount of neutral oligosaccharides in colostrum does not directly correlate with maternal allergy, nor with allergy development in children up to 18 months of age.     

Oligosaccharides might stimulate calcium absorption in formula-fed preterm infants

Among other components of human milk, oligosaccharides might contribute to the high efficiency of calcium absorption of breastfed infants. In adults, it can be shown that dietary oligosaccharides can improve calcium absorption. The present analysis was performed to evaluate a possible influence of dietary oligosaccharides on parameters of calcium metabolism in preterm infants. The concentrations of calcium and phosphorus in plasma and in spot urine samples as well as the plasma activity of the alkaline phosphatase were measured in preterm infants fed either a standard formula ( n = 15) or a formula supplemented with dietary oligosaccharides ( n = 15) at the end of a 4-wk feeding period.
Conclusions : There was no influence of the different diets on the plasma concentration of calcium and phosphorus or on the plasma activity of alkaline phosphatase. In urine, there was a tendency towards higher calcium concentrations in the group fed the supplemented formula compared concentrations in the group fed the standard formula. The concentrations of phosphate were not significantly different and, as a consequence, there was a tendency towards a higher Ca/P molar ratio in the group fed the supplemented formula. The data indicate that the calcium absorption might be influenced by the dietary oligosaccharides. Thus, the possible effect of dietary oligosaccharides on calcium homeostasis should be included in the discussion concerning the consequences of the use of dietary oligosaccharides in preterm infant nutrition.     

Affinity chromatographic identification and quantitation of blood group A-active oligosaccharides in human milk and feces of breast-fed infants

The finding of large quantities of blood group A-active oligosaccharides in the feces of a blood group A breast-fed infant motivated a search for the origin of these compounds. Using an affinity chromatographic technique, the nature of A-active oligosaccharides in human milk is demonstrated. The amounts of A-active tetrasaccharide (A-tetra) and the Lewis b-active lacto-N-difucohexaose I (LND-I) varied between 19-375 mg/L for A-tetra and 14-710 mg/L for LND-I. Using the same technique, the amounts of A-tetra and LND-I in milk samples from five women of different blood groups were compared with those in the feces of their breast-fed infants. The A-tetra was present only in feces from infants of blood group A or AB mothers and the amount per 24 h corresponded roughly to that in a I-L portion of milk. One of the milk samples was also analyzed for the presence of larger A-active oligosaccharides (A-pentasaccharide, A-hexasaccharide, and A-heptasaccharide). Their amounts were much less as compared to the amounts present in feces. These results indicate that milk is a possible source for the smallest A-tetrasaccharide found in the feces of breast-fed infants, while the larger A-active oligosaccharides might be the result of an intestinal metabolic modification.     

Oligosaccharides might stimulate calcium absorption in formula-fed preterm infants

Among other components of human milk, oligosaccharides might contribute to the high efficiency of calcium absorption of breastfed infants. In adults, it can be shown that dietary oligosaccharides can improve calcium absorption. The present analysis was performed to evaluate a possible influence of dietary oligosaccharides on parameters of calcium metabolism in preterm infants. The concentrations of calcium and phosphorus in plasma and in spot urine samples as well as the plasma activity of the alkaline phosphatase were measured in preterm infants fed either a standard formula (n = 15) or a formula supplemented with dietary oligosaccharides (n = 15) at the end of a 4-wk feeding period. CONCLUSIONS: There was no influence of the different diets on the plasma concentration of calcium and phosphorus or on the plasma activity of alkaline phosphatase. In urine, there was a tendency towards higher calcium concentrations in the group fed the supplemented formula compared concentrations in the group fed the standard formula. The concentrations of phosphate were not significantly different and, as a consequence, there was a tendency towards a higher Ca/P molar ratio in the group fed the supplemented formula. The data indicate that the calcium absorption might be influenced by the dietary oligosaccharides. Thus, the possible effect of dietary oligosaccharides on calcium homeostasis should be included in the discussion concerning the consequences of the use of dietary oligosaccharides in preterm infant nutrition.     

Oligosaccharides in colostrum of Italian and Burkinabe women

Human milk contains a large number of compounds to provide nutrition and defense for the newborn. Among these, oligosaccharides are present in concentrations up to 12 g/L, and their composition varies during lactation. Colostrum from 53 Burkinabe women were collected at the maternity department of St Camille Medical Centre in Ouagadougou (Burkina Faso, West Africa). Colostrum from 50 Italian women were collected at the maternity department of St Bambino Hospital in Catania (Catania, East Sicily, Italy). All mothers spontaneously delivered at term. Italian mothers received an injection of the ergot derivative ergotamine after delivery. Ergotamine, notoriously, delays breastfeeding initiation up to 2 to 3 days. Chromatographic separation of colostrum from both Burkinabe and Italian women showed a progressive appearance of oligosaccharides in the first 3 days. Burkinabe women showed high concentrations of 2-fucosyllactose and lower concentrations of lacto-N-fucopentaose I. By contrast, Italian women showed inverted behaviour. A comparable percentage of the secretor genotype for the Lewis blood group phenotype in both Burkinabe and Italian women was found. According to the different ethnicity, different milk oligosaccharide profiles were documented in the present study. 2-Fucosyllactose in milk should be biologically significant for Burkinabe infants because of the high levels found in their mothers' colostrum after the second day of lactation.     

Prebiotic carbohydrates in human milk and formulas

Human milk oligosaccharides play an important role, as prebiotic soluble fibres, in the postnatal development of the intestnial flora. Infant formulas are virtually free of prebiotic oligosaccharides. As a consequence, formula-fed infants develop an intestinal flora significantly different to the flora of breastfed infants. Due to the complexity of human milk oligosaccharides, it is necessary to use alternative sources of prebiotic ingredients as components of infant formulas. The present review summarizes the data of experimental research and clinical studies with a prebiotic mixture containing 90% short-chain galacto-oligosaccharides and 10% long-chain fructo-oligosacchrides are summarized. The data demonstrate that, with this prebiotic mixture, the growth of bifidobacteria and lactobacilli can be stimulated, the faecal pH can be decreased, and the presence of pathogens can be reduced to levels similar to those of breastfed infants. Thus, prebiotic oligosaccharides such as the studied mixture provide beneficial effects for formula-fed infants.     

Dietary prebiotic oligosaccharides are detectable in the faeces of formula-fed infants

Human milk oligosaccharides are not digested during intestinal passage and can be detected in stools. In this study it was investigated whether a prebiotic mixture of low-molecular-weight galacto-oligosaccharides (GOS) and high-molecular-weight fructo-oligosaccharides (FOS) can be detected in stool samples of formula-fed infants. The test formula was supplemented with 0.8 g/dl oligosaccharides (GOS+FOS). In the control formula, maltodextrins were used as placebo. Fecal flora was assessed at the beginning (day 1) and at the end of a 28-d feeding period (day 2). At day 2 the content of galacto- and fructo-oligosaccharides in the stool samples were measured. On study day 1, the number of bifidobacteria was not different among the groups (supplemented group: 7.7 (6.2) CFU/g; placebo group: 8.0 (6.0) CFU/g). At the end of the 28-d feeding period, the number of bifidobacteria was significantly higher in the group fed the supplemented formula when compared to placebo (supplemented group: 9.8 (0.7) CFU/g stool; placebo group: 7.1 (4.7) CFU/g stool; p <0.001). In all infants fed the supplemented formula, GOS and FOS could be identified in the stool samples. That was not the case in infants fed the non-supplemented formula.
Conclusion: The present data confirm the bifidogenicity of oligosaccharides and indicate that dietary galacto-oligosaccharides and long chain fructo-oligosaccharides remain during the whole passage in the lumen of the gastrointestinal tract, similarly to human milk oligosaccharides.     

Dietary prebiotic oligosaccharides are detectable in the faeces of formula-fed infants

Human milk oligosaccharides are not digested during intestinal passage and can be detected in stools. In this study it was investigated whether a prebiotic mixture of low-molecular-weight galacto-oligosaccharides (GOS) and high-molecular-weight fructo-oligosaccharides (FOS) can be detected in stool samples of formula-fed infants. The test formula was supplemented with 0.8 g/dl oligosaccharides (GOS+FOS). In the control formula, maltodextrins were used as placebo. Fecal flora was assessed at the beginning (day 1) and at the end of a 28-d feeding period (day 2). At day 2 the content of galacto- and fructo-oligosaccharides in the stool samples were measured. On study day 1, the number of bifidobacteria was not different among the groups (supplemented group: 7.7 (6.2) CFU/g; placebo group: 8.0 (6.0) CFU/g). At the end of the 28-d feeding period, the number of bifidobacteria was significantly higher in the group fed the supplemented formula when compared to placebo (supplemented group: 9.8 (0.7) CFU/g stool; placebo group: 7.1 (4.7) CFU/g stool; p<0.001). In all infants fed the supplemented formula, GOS and FOS could be identified in the stool samples. That was not the case in infants fed the non-supplemented formula. CONCLUSION: The present data confirm the bifidogenicity of oligosaccharides and indicate that dietary galacto-oligosaccharides and long chain fructo-oligosaccharides remain during the whole passage in the lumen of the gastrointestinal tract, similarly to human milk oligosaccharides.     

Prebiotic carbohydrates in human milk and formulas

Human milk oligosaccharides play an important role, as prebiotic soluble fibres, in the postnatal development of the intestinal flora. Infant formulas are virtually free of prebiotic oligosaccharides. As a consequence, formula-fed infants develop an intestinal flora significantly different to the flora of breastfed infants. Due to the complexity of human milk oligosaccharides, it is necessary to use alternative sources of prebiotic ingredients as components of infant formulas. The present review summarizes the data of experimental research and clinical studies with a prebiotic mixture containing 90% short-chain galacto-oligosaccharides and 10% long-chain fructo-oligosacchrides are summarized. The data demonstrate that, with this prebiotic mixture, the growth of bifidobacteria and lactobacilli can be stimulated, the faecal pH can be decreased, and the presence of pathogens can be reduced to levels similar to those of breastfed infants. Thus, prebiotic oligosaccharides such as the studied mixture provide beneficial effects for formula-fed infants.