Objective: To explore and establish the preparation system of human intestinal fluid transplantation (HIFT) and HIFT capsule, and to preliminarily apply it to clinic. Methods: Strict standards for donor screening and management were established. The nasojejunal tube was catheterized into the distal jejunum, and then it was connected with an improved disposable sterile negative pressure collection device for the collection of human intestinal fluid. After that, it was prepared into capsules by filtering, adding 10% glycerin protectant and freeze-drying method. The amount of living bacteria was used as the standard of therapeutic dose. The living bacteria amount in fluid is ≥ 5.0×108 /mL and the living bacteria proportion is ≥ 83%; the living bacteria amount in powder is ≥ 2.0×106 /g and the living bacteria proportion is ≥ 81%; The observational indicators included: (1) the basic information of the donor, the amount of living bacteria in the HIF and powder. (2) Preliminary analysis of the treatment for ASD, which combined HIFT capsule with standard FMT capsule, from February to December 2021 (Clinical trial Registration Number: ChiCTR2100043929). Evaluation criteria: Trypan blue staining method was used to detect the living bacteria amount in fluid and powder. The Autism Behavior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were used to evaluate the efficacy. Results: Compared with the parent donor, the standard donor was younger [(25.4±0.9) y vs. (30.7±3.2) y, t=-19.097, P=0.001] and had a lower body mass index [(19.7±0.5) kg/m2 vs. (20.8±1.3) kg/m2, t=-8.726, P=0.001], more in the living bacteria amount in powder [(7.47±1.52)×106/g vs. (5.03±1.38)×106/g, t=11.331, P=0.031], Chao index (205.4±6.8 vs. 194.2±7.2, t=10.415, P=0.001), and Shannon index (3.25±0.14 vs 2.72±0.27, t=19.465, P=0.001). The differences were statistically significant (all P<0.05). However, there were no significant differences in gender, drainage volume and total number of bacterial liquid colonies between the two groups (all P>0.05). Both the standard donor and the parent donor met the donor screening criteria, and the preparation fluid and powder met the treatment criteria. Eight patients received the treatment of HIFT combined with fecal microbiota transplantation (FMT). Preliminary statistical results showed that HIFT combined with FMT improved ABC and CARS at the 1st, 2nd, 3rd and 4th months. The differences were statistically significant (all P<0.05). No severe adverse reaction occurred. Conclusion: Based on the previous research on FMT preparation system and the clinical technology in our center, this study developed a high standard HIFT preparation system, and explored the clinical study of HIFT combined with FMT, in order to provide an innovative therapy for the treatment of diseases.
目的： 探索建立标准化人体小肠液移植（HIFT）及HIFT胶囊的制备体系，并分析其初步应用于孤独症谱系障碍的治疗效果。 方法： 收集2021年1月至3月于上海市第十人民医院招募的HIFT标准供体3人、孤独症谱系障碍患者亲体供体8人的临床资料。通过制定严格的供体筛选与管理标准，运用床旁经鼻空肠管置管术，将导管头端置入空肠远端，导管连接口处接改良一次性无菌负压收集装置行持续负压引流收集人体小肠液。再分别通过过滤、添加10%甘油保护剂及冻干等方法制成冻干粉HIFT胶囊。本中心初步拟定要求：移植以活菌量作为治疗剂量标准，HIFT每次移植的菌液活菌量需≥5.0×108个/ml且活菌比例≥83%，菌粉活菌量需≥2.0×106个/g且活菌比例≥81%。观察供体基本情况、供体中菌液及菌粉中菌落总数，并初步分析上述供体制备而成的HIFT胶囊联合标准FMT胶囊治疗（即全肠道菌群移植）治疗孤独症谱系障碍研究（临床试验注册号：ChiCTR2100043929）。 结果： 标准供体及亲体供体均符合供体筛选标准，所制菌液及菌粉符合治疗标准。与亲体供体比较，标准供体的菌粉菌落总数更多［（7.47±1.52）×106个/g比（5.03±1.38）×106个/g，t=11.331，P=0.031］、Chao指数更高（205.4±6.8比194.2±7.2，t=10.415，P=0.001）、Shannon指数更高（3.25±0.14比2.72±0.27，t=19.465，P=0.001），差异均具有统计学意义（均P<0.05）。有8例患者接受全肠道菌群移植治疗，初步统计结果显示，该疗法可改善孤独症谱系障碍患者症状，治疗后1、2、3、4个月时，孤独症行为评分及儿童孤独症评分均下降，差异均有统计学意义（均P<0.05）。本组无严重不良反应发生。 结论： 在FMT制备体系研究基础上，结合制定高标准HIFT制备体系，同时探索性开展HIFT联合FMT的全肠道菌群移植临床研究，有望成为微生态治疗的一项革新疗法。.