Objective: To investigate the influence of family integrated care (FICare) on the intestinal microbiome of preterm infants in neonatal intensive care unit (NICU). Methods: This was a prospective observational pilot study. A total of 44 preterm infants (23 boys, 52%) admitted to NICU of the Third Xiangya Hospital of Central South University from July, 2015 to June, 2017 were enrolled and divided into FICare, non-FICare groups. Totally 20 term infants (11 boys, 55%) were enrolled into control group, who were sent to the Pediatric Healthcare Clinic for regular health check on postnatal 28-31 days. All infants were free from probiotics after birth and on full enteral feeding. Clinical data of all infants were collected. Two fresh stool specimens of infants in FICare group were collected after 2 weeks of FICare implementation, without use of antibiotics during the prior 1 week. Stool specimens of infants in non-FICare group were collected at the meantime;while for the infants in control group, stool samples were collected at 4 weeks of age. All specimens were stored in-80 ℃ freezer, subsequently investigated by 16 S rRNA sequencing. The results were filtered by paired-end reads software based on RNA overlapping-splicing and tags calculation. Operational taxonomic units (OTU) were analyzed for intestinal microbiome richness. Intestinal microbiome diversity was measured with Shannon index. One-way ANOVA or Kruskal-Wallis H statistic analysis or Chi-square test was used for statistical analysis. Results: There were no significant differences among FICare, non-FICare and control groups in male proportion (52% (11/21) vs. 52% (12/23) vs. 55% (11/20), χ(2)=0.041, P=0.980), in-born ratio (90% (19/21) vs. 87% (20/23) vs. 85% (17/20), χ(2)=0.000, P=1.000), and percentage of infants with Apgar scores<7 at 5 minutes after birth (14% (3/21) vs. 9% (2/23) vs. 5% (1/20), χ(2)=0.120, P=0.729). Similarly, no significant differences were found between FICare and non-FICare groups in terms of gestational age ((29.7±1.8) vs. (29.9±1.7) weeks, t=0.378, P=0.707), birth weight ((1 266±310) vs. (1 326±318) g, t=0.631, P=0.531), median age of initiating feeds (4 vs. 4 days old, Z=0.666, P=0.505), and median age of achieving feeding volume of 120 ml/(kg·d)(13 vs. 11 days old, Z=1.014, P=0.310). However, the breast-feeding rate in FICare group (18/21, 86%) was significantly higher than that in non-FICare group (8/23, 35%) (t=11.780, P=0.001). The medium Shannon index was 0.72 (0.27,2.66), 0.61 (0.18,1.83), and 0.52 (0.08,1.71) in control, FICare, and non-FICare groups, respectively, without significant difference (H=1.823, P=0.402). The domain flora was Lactobacillus Firmicutes in all three groups, which was of the highest percentage in FICare group (71.6±5.4)%, followed by control group (65.4±6.6)% and non-FICare group (55.6±8.8)%, with a significant difference (F=27.919, P=0.000). Conclusions: FICare can improve the richness and diversity of intestinal microbiome, stimulate the establishment of flora close to those of normal breast-feeding infants in preemies in NICU, making its establishment being more similar to normal term breast-feeding infants. This effect might be caused by the increased skin-to-skin contact and increased fresh breast-milk-feeding in FICare.
目的： 探讨家庭参与式综合管理（FICare）对新生儿重症监护病房（NICU）住院早产儿肠道菌群的影响。 方法： 采用前瞻性、观察性研究，研究对象为2015年7月至2017年6月在中南大学湘雅三医院NICU住院的早产儿44例（其中男23例，占52%）分为FICare组和非FICare组，同期儿童保健门诊满月体检的足月儿20名（其中男11名，占55%）作为对照组。所有新生儿均为全肠内喂养且出生后未接触肠道益生菌，收集临床数据。FICare组于在FICare超过2周且未接触抗菌药物1周时留取双份新鲜粪便，非FICare组与FICare组在相对应的时间点、对照组在新生儿4周龄时留取双份新鲜粪便，-80 ℃冰冻保存，标本统一进行高通量16 S rRNA肠道菌群高通量测序。应用Paired End Reads肠道菌群RNA读取软件对数据经过质量过滤，统计各样品操作分类单元（OTU）的丰度信息，以Shannon指数作为肠道菌群多样性的指标。采用单因素方差分析或非参数检验的Kruskal-Wallis H法及连续校正χ(2)检验或似然比χ(2)检验进行组间统计学比较。 结果： FICare组、非FICare组和对照组的男婴比例[52%（11/21）比52%（12/23）比55%（11/20）]、本院出生比例[90%（19/21）比87%（20/23）比85%（17/20）]及5 min Apgar评分<7分比例[14%（3/21）比9%（2/23）比5%（1/20）]的差异均无统计学意义（χ(2)=0.041、0.000、0.120，P=0.980、1.000、0.729）。FICare组与非FICare组的出生胎龄、出生体重、中位开奶日龄、奶量达120 ml/（kg·d）中位日龄差异均无统计学意义[（29.7±1.8）比（29.9±1.7）周、（1 266±310）比（1 326±318）g、4比4日龄、13比11日龄，t=0.378、0.631，Z=0.666、1.014，P=0.707、0.531、0.505、0.310]。FICare组与非FICare组的母乳喂养率分别为86%（18/21）比35%（8/23），差异有统计学意义（t=11.780，P=0.001）。对照组、FICare组、非FICare组新生儿的肠道菌群多样性Shannon指数分别为0.72（0.27，2.66）、0.61（0.18，1.83）、0.52（0.08，1.71），差异无统计学意义（H=1.823，P=0.402）。FICare组、非FICare组和对照组新生儿肠道菌群以厚壁菌门的乳酸杆菌属为主，乳酸杆菌属占比分别为（71.6±5.4）%、（55.6±8.8）%和（65.4±6.6）%，差异有统计学意义（F=27.919，P=0.000）。 结论： FICare可提高NICU住院早产儿的肠道菌群丰富度和多样性，促进肠道菌群的建立，使其更接近于正常足月母乳喂养儿肠道菌群分布，但该作用可能是FICare为母子皮肤接触、新鲜亲母抱喂提供了更多机会而导致的。.
Keywords: Family integrated care; High-throughput nucleotide sequencing; Infant, premature; Intensive care units, neonatal; Intestinal microbiome.