We have previously reported on the potential pathogenic role of neutrophils in biliary atresia (BA). Herein, we aimed to delineate the role of CD177+ neutrophils in the pathogenesis of BA. Immune cells from the livers of mice with rhesus rotavirus-induced BA were analysed. Single-cell RNA-sequencing was performed to specifically analyse Gr-1⁺ (Ly6C/Ly6G⁺) cells in the liver. Gene expression profiles of CD177⁺ cells were analysed using the Smart-Seq RNA-sequencing method, and the pathogenesis of BA was examined in Cd177^-/- mice. Neutrophil extracellular trap (NET) inhibitors were used to determine the role of CD177⁺ cell-derived NETs in BA-associated bile duct damage, and a pilot clinical study evaluated the potential effects of N-acetylcysteine on NET release in BA. Increased levels of Gr-1⁺ cells were observed in the livers of mice with rhesus rotavirus-induced BA. RNA-sequencing analysis revealed that CD177⁺ cells were the main population of Gr-1⁺ cells and expressed elevated levels of both interferon-stimulated and neutrophil degranulation genes. Cd177^-/- BALB/c mice exhibited delayed disease onset and reduced morbidity and mortality. High numbers of mitochondria were detected in CD177⁺ cells derived from mice with BA; these cells were associated with increased levels of reactive oxygen species and increased NET formation, which induced the apoptosis of biliary epithelial cells in cocultures. In a pilot clinical study, the administration of N-acetylcysteine to patients with BA reduced CD177⁺ cell numbers and reactive oxygen species levels, indicating a potential beneficial effect. Our data indicate that CD177⁺ cells play an important role in the initiation of BA pathogenesis via NET formation. The pilot study of N-acetylcysteine treatment in patients with BA was registered on the Chinese Clinical Trial Registry (ChiCTR2000040505). Neutrophils (a type of innate immune cell, i.e. an immune cell that doesn't target a specific antigen) are thought to play a role in the development of biliary atresia (a rare but potentially lethal condition of the bile ducts that occurs in infants). Herein, we found that neutrophils expressing a particular protein (CD177) played an important role in bile duct damage by releasing a special structure (NET) that can trap and kill pathogens but that can also cause severe tissue damage. A pilot study in patients with biliary atresia showed that inhibiting NETs could have a beneficial effect.

Biliary atresia (BA) is a multifactorial pathogenic disease with possible genetic components. As a member of membrane skeletal proteins in the liver and bile ducts, a haplotype composed by five single nucleotide polymorphisms (SNPs) on adducin 3 (ADD3) has been identified as associated with BA. However, limited study was designed to further elaborate the mutual relationship amongst those replicated SNPs to disease. We selected three susceptibility SNPs in ADD3 and conducted a replication study using 510 BA cases and 1473 controls to evaluate the individual function of the SNPs and further stratified the potential roles with disease and its subclinical features. Two SNPs in ADD3 were replicated as associated with BA (1.60E-04 ≤ P≤1.70E-04, 1.33 ≤ odds ratio (OR) ≤ 1.58 for rs17095355, 2.10E-04 ≤ P≤5.30E-04, 1.26 ≤ OR ≤ 1.57 for rs2501577). Though we failed to replicate the individual association of rs10509906 to disease, the intragenic epistatic effect between rs10509906 and rs2501577 was suggested as exhibiting susceptibility to BA, further cross-validated by multifactor dimensionality reduction (MDR) (P=0.068, OR = 1.37), which may explain extra hidden heritability of ADD3 to BA. Furthermore, through subclinical stratification, we also observed the association of risk to disease mainly came from the female patients.

Background: To analyze the early outcome of thoracoscopic repair (TR) of neonatal congenital diaphragmatic hernia (CDH) with standardized indications. Methods: Clinical data of 14 neonates with CDH who underwent TR from September 2013 to August 2014 were retrospectively analyzed. Selection criteria: weight beyond 2.0 kg; liver in the abdomen; no intestinal obstruction; no severe cardiopulmonary anomalies; not required high-frequency oscillatory ventilation or extracorporeal membrane oxygenation. Timing of surgery: mean arterial blood pressure normal for gestational age; preductal saturation levels of 85%–95% on fractional inspired oxygen below 50%; lactate below 3 mmol/L; urine output more than 2 mL/kg/h. Fourteen cases in the historical control group who underwent open repair (OR) with the same physiological status were reviewed for comparison. Results: Demographic features were similar between the TR group and OR group. The TR group had a higher intraoperative mean PaCO₂ (48 ± 8 mmHg versus 39 ± 6 mmHg, P = .0024) and mean arterial pH (7.30 ± 0.06 versus 7.39 ± 0.06, P = .0005), but no differences in lactate (0.93 ± 0.16 mmol/L versus 0.98 ± 0.14 mmol/L, P = .3869). Longer operation time was found in the TR group (116 ± 27 minutes versus 74 ± 25 minutes, P = .0002). No recurrence was observed in groups within the first year of life follow-up. Conclusions: With selection criteria and timing, TR of CDH in neonates can be performed safely and successfully.