Antimicrobial peptide LL37 and regulatory T cell associated with late-onset sepsis in very preterm infants.

Stem cell therapy may prevent late-onset sepsis (LOS) via antimicrobial peptide LL37 secretion and regulatory T cell (Treg) regulation. The early prediction of LOS is still a challenge. This study evaluated whether immunological state of LL37 or Tregs precedes LOS. We firstly analyzed the LL37 level, Treg proportion, and LOS incidence in very preterm infants treated with autologous cord blood mononuclear cells (ACBMNCs) in our previous trial. Then, we constructed a prediction model and built validation cohort. We found ACBMNC intervention reduced the incidence of LOS from 27.3% to 6.9% (p = 0.021). LL37 and Treg abundances were higher in the ACBMNCs group. The nomogram demonstrated that early-life Treg and LL37 characteristics were closely associated with LOS (area under the curve, AUC 0.936), with implications for early prediction and timely clinical management. This composite model was also helpful to evaluate the beneficial effect of ACBMNCs intervention on LOS, thus promoting translational research.

Umbilical cord blood cell characteristics in very preterm neonates for autologous cell therapy of preterm-associated complications.

BACKGROUND: There are emerging clinical evidence for umbilical cord blood mononuclear cells (UCBMNCs) intervention to improve preterm complications. The first critical step in cell therapy is to obtain high-quality cells. This retrospective study aimed to investigate the quantity and quality of UCBMNCs from very preterm infants (VPIs) for the purpose of autologous cell therapy in prevention and treatment of preterm complications. METHODS: Very preterm infants (VPIs) born in Guangdong Women and Children Hospital from January 1, 2017, to December 8, 2022, from whom cord blood was successfully collected and separated for public or private banking, were enrolled. The UCBMNCs characters from route cord blood tests performed in cord blood bank, impact of perinatal factors on UCBMNCs, the relationship between UCBMNCs characteristics and preterm outcomes, and the correlation of UCBMNCs characteristics and peripheral blood cells in VPIs were analyzed. RESULTS: Totally, 89 VPIs underwent UCB collection and processing successfully. The median cell number post processing was 2.6 × 108. To infuse a dose of 5 × 107 cells/kg, only 3.4% of infants required a volume of more than 20 mL/kg, which exceeded the maximum safe volume limit for VPIs. However, when infusing 10 × 107 cells/kg, 25.8% of infants required a volume of more than 20 ml/kg volume. Antenatal glucocorticoids use and preeclampsia was associated with lower original UCBMNCs concentration. Both CD34+ hematopoietic stem cells (HSC) frequency and colony forming unit - granulocyte and macrophage (CFU-GM) number correlated negatively with gestational age (GA). UCBMNCs characters had no significant effect on preterm outcomes, whereas a significant positive correlation was observed between UCBMNCs concentration and total white blood cell, neutrophil, lymphocyte and PLT counts in peripheral blood. CONCLUSION: UCBMNCs collected from VPIs was feasible for autologous cell therapy in improving preterm complications. Setting the infusion dose of 5 × 107 cells/kg guaranteed a safe infusion volume in more than 95% of the targeted infants. UCBMNCs characters did not affect preterm complications; however, the effect of UCBMNCs concentration on peripheral blood classification count should be considered when evaluating the immunomodulation of UCBMNCs transfusion.

Prevention for moderate or severe BPD with intravenous infusion of autologous cord blood mononuclear cells in very preterm infants-a prospective non-randomized placebo-controlled trial and two-year follow up outcomes.

Background: Bronchopulmonary dysplasia (BPD) is the primary severe complication of preterm birth. Severe BPD was associated with higher risks of mortality, more postnatal growth failure, long term respiratory and neurological developmental retardation. Inflammation plays a central role in alveolar simplification and dysregulated vascularization of BPD. There is no effective treatment to improve BPD severity in clinical practice. Our previous clinical study showed autologous cord blood mononuclear cells (ACBMNCs) infusion could reduce the respiratory support duration safely and potential improved BPD severity. Abundant preclinical studies have reported the immunomodulation effect as an important mechanism underlying the beneficial results of stem cell therapies in preventing and treating BPD. However, clinical studies assessing the immunomodulatory effect after stem cells therapy were rare. This study was to investigate the effect of ACBMNCs infusion soon after birth on prevention for severe BPD and long term outcomes in very preterm neonates. The immune cells and inflammatory biomarkers were detected to investigate the underlying immunomodulatory mechanisms. Methods: This single-center, prospective, investigator-initiated, non-randomized trial with blinded outcome assessment aimed to assess the effect of a single intravenous infusion of ACBMNCs in preventing severe BPD (moderate or severe BPD at 36 weeks of gestational age or discharge home) in surviving very preterm neonates less than 32 gestational weeks. Patients admitted to the Neonatal Intensive Care Unit (NICU) of Guangdong Women and Children Hospital from July 01, 2018 to January 1, 2020 were assigned to receiving a targeted dosage of 5 × 107 cells/kg ACBMNC or normal saline intravenously within 24 h after enrollment. Incidence of moderate or severe BPD in survivors were investigated as the primary short term outcome. Growth, respiratory and neurological development were assessed as long term outcomes at corrected age of 18-24 month-old. Immune cells and inflammatory biomarkers were detected for potential mechanism investigation. The trial was registered at ClinicalTrials.gov (NCT02999373). Findings: Six-two infants were enrolled, of which 29 were enrolled to intervention group, 33 to control group. Moderate or severe BPD in survivors significantly decreased in intervention group (adjusted p = 0.021). The number of patients needed to treat to gain one moderate or severe BPD-free survival was 5 (95% confidence interval: 3-20). Survivors in the intervention group had a significantly higher chance to be extubated than infants in the control group (adjusted p = 0.018). There was no statistical significant difference in total BPD incidence (adjusted p = 0.106) or mortality (p = 1.000). Incidence of developmental delay reduced in intervention group in long term follow-up (adjusted p = 0.047). Specific immune cells including proportion of T cells (p = 0.04) and CD4+ T cells in lymphocytes (p = 0.03), and CD4+ CD25+ forkhead box protein 3 (FoxP3)+ regulatory T cells in CD4+ T cells increased significantly after ACBMNCs intervention (p ï¼œ 0.001). Anti-inflammatory factor IL-10 was higher (p = 0.03), while pro-inflammatory factor such as TNF-a (p = 0.03) and C reactive protein (p ï¼œ 0.001) level was lower in intervention group than in control group after intervention. Interpretation: ACBMNCs could prevent moderate or severe BPD in surviving very premature neonates and might improve neurodevelopmental outcomes in long term. An immunomodulatory effect of MNCs contributed to the improvement of BPD severity. Funding: This work was supported by National Key R&D Program of China (2021YFC2701700), National Natural Science Foundation of China (82101817, 82171714, 8187060625), Guangzhou science and technology program (202102080104).

Wnt5a-Flt1 activation contributes to preterm altered cerebral angiogenesis after prenatal inflammation.

OBJECTIVE: Intraventricular hemorrhage (IVH) causes morbidity and mortality in preterm infants and prenatal exposure to inflammation contributes to brain injury. Moreover, prenatal exposure to severe inflammation increases the risk of IVH in preterm neonates. The current study investigated whether intrauterine exposure to inflammation affects cerebral angiogenesis and its underlying mechanisms. METHODS: Wnt5a, flt1, and vascular endothelial growth factor (VEGF)-A levels in cord blood serum (stored in a bio-bank) of the enrolled patients were measured via enzyme-linked immunosorbent assay. A preterm prenatal inflammation exposure model was established in rats by intraperitoneal injection intraperitoneally during pregnancy. Angiogenesis of cerebral tissue was analyzed using immunohistochemistry. Wnt5a, flt1, and VEGF-A expression levels were measured via immunohistochemistry, immunofluorescence, or western blotting. The correlation between Wnt5a and flt1 expression and the cerebral vessel area was also analyzed. RESULTS: The Wnt5a and flt1 levels in the cord blood serum were significantly higher in the amnionitis group than in the non-amnionitis group. The VEGF-A level in the cord blood serum was significantly lower in the amnionitis group. In the rat model, preterm rats in the prenatal inflammation group exhibited increased microglial cell infiltration and decreased vessel area and diameter in the cerebral tissue compared to the control group. Wnt5a was located in microglial cells, and Wnt5a and flt1 expression in brain tissue significantly increased after prenatal lipopolysaccharide (LPS) exposure. VEGF-A expression declined after prenatal LPS exposure. The cerebral vessel area was negatively correlated with Wnt5a and flt1 expression. CONCLUSION: Disordered cerebral angiogenesis is associated with increased Wnt5a-Flt1 activation in microglial cells after exposure to intrauterine inflammation.

Autologous cord blood mononuclear cell infusion for the prevention of bronchopulmonary dysplasia in very preterm monozygotic twins: A study protocol for a randomized, placebo-controlled, double-blinded multicenter trial.

Background: Preterm-associated complications remain the main cause of neonatal death. Survivors face the challenges of short- and long-term complications. Among all complications, bronchopulmonary dysplasia (BPD) remains the first important cause of neonatal mortality and morbidity. Current treatment does not address this main preterm complication. Cord blood is regarded as a convenient source of stem cells. The paracrine bioactive factors of stem cells contribute to tissue repair and immune modulation. Our clinical studies and those of others have shown that cord blood cell infusion is both safe and possibly effective in the prevention and treatment of BPD. The therapeutic use of cord blood has emerged as a promising therapy. However, the genetic heterogeneity between control and intervention groups may reduce the comparability especially among small sample trials. The purpose of this study protocol is to investigate the effects of autologous cord blood mononuclear cell (ACBMNC) infusion on the prevention of BPD in very preterm monozygotic twins of less than 32 gestation weeks. Methods: In this prospective, randomized, placebo-controlled, double-blinded multicenter clinical trial, 60 pairs of monozygotic twin preterm neonates of less than 32 weeks admitted to the Neonatal Intensive Care Unit are randomly assigned to receive intravenous ACBMNC infusion (targeted at 5 × 107 cells/kg) or placebo (normal saline) within 24 h after birth in a 1:1 ratio. The primary outcome will be survival without BPD at 36 weeks of postmenstrual age. The secondary outcomes will include the mortality rate, BPD severity, other common preterm complication rates, respiratory support duration, length and cost of hospitalization, and long-term respiratory and neurodevelopmental outcomes during a 2-year follow-up. Furthermore, we will perform single-cell RNA sequencing for cord blood cells and blood cells 3-10 days after intervention and detect whether reactive oxygen species and inflammatory cytokines are present. Conclusion: This will be the first randomized, placebo-controlled, double-blinded trial to evaluate the efficacy of ACBMNC infusion to prevent BPD in monozygotic twin premature infants and investigate the underlying protective mechanisms. The results of this trial will provide valuable clinical evidence for translational application of cord blood cell therapy in very preterm infants.Trial registration: ClinicalTrials.gov, NCT05087498, registered 10/09/2021, https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S000BAD7&selectaction=Edit&uid=U0002PLA&ts=2&cx=qvyylv.

The correlation between chorioamnionitis-induced abnormal placental microvessel and platelet metabolism of preterm infants / 中华实用儿科临床杂志

Objective@#To observe the effect of chorioamnionitis on placental microvessel and platelet metabolism in premature and the correlation between them.@*Methods@#With clinical randomized controlled trial (RCT), the cases were matched by 11 according to gestational ages and divided into 2 groups according to the placental pathology results: chorioamnionitis group and control group, 32 cases in each group.Dates were obtained for preterm infants (gestational age<37 weeks) admitted to the Department of Neonatology at Guangdong Women and Children Hospital, born between June and December 2016.The platelet parameter [platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT)], birth weight, thrombocytopenia, hemorrhage complication, miscrovascular density (MVD) in placenta, platelet activating factors (CD62p, CD63) and thrombopoietin (TPO) in preterm infants were recorded and compared.@*Results@#In chorioamnionitis group, the infant birth weight[(1.90±0.41) kg vs.(2.31±0.62) kg] and the PLT in 72 hours [<24 h (197.97±63.43)×109/L vs.(266.34±69.92)×109/L; 24-72 h (202.28±29.70)×109/L vs.(256.38±69.96)×109/L] were significantly lower compared with the control group, and the differences were statistically significant(all P<0.05). The incidence of early thrombocytopenia(37.50% vs.9.38%), intracranial hemorrhage(40.62% vs.15.63%), MPV [(8.73±0.89) fL vs.(8.27±0.64) fL] and PDW[(59.46±5.90)% vs.(55.20±5.37)%] in 24 hours were significantly higher in chorioamnionitis group, and the differences were statistically significant (all P<0.05). The placental MVD in chorioamnionitis group significantly decreased[(9.08±1.35)% vs.(12.89±1.36)%, P<0.05 ]. The level of CD62p, CD63 and TPO in umbilical cord blood were significantly higher in chorioamnionitis group[(25.37±5.20)% vs.(10.35±2.94)%, (9.49±1.58)% vs.(4.04±1.21)%, (271.08±197.22) μg/L vs.(141.87±78.10) μg/L, all P<0.05]. The placental MVD was positively associated with PLT (r=0.74, P<0.05) and negatively associated with CD62p, CD63 and TPO among infants with chorioamnionitis (r=-0.64, -0.44, -0.44, all P<0.05).@*Conclusions@#The chorioamnionitis may decrease the MVD in placenta and activate platelet in fetal circulation, damaged placental microvessel may activate platelet further.

The correlation betWeen chorioamnionitis＿induced abnormal placental microvessel and platelet metabolism of preterm infants / 中华实用儿科临床杂志

Objective To observe the effect of chorioamnionitis on placental microvessel and platelet metabo-lism in premature and the correlation between them. Methods With clinical randomized controlled trial( RCT),the cases were matched by 1: 1 according to gestational ages and divided into 2 groups according to the placental pathology results:chorioamnionitis group and control group,32 cases in each group. Dates were obtained for preterm infants(ges-tational age<37 weeks)admitted to the Department of Neonatology at Guangdong Women and Children Hospital,born between June and December 2016. The platelet parameter[platelet count(PLT),mean platelet volume(MPV),plate-let distribution width( PDW),plateletcrit( PCT)],birth weight,thrombocytopenia,hemorrhage complication,miscro-vascular density(MVD)in placenta,platelet activating factors( CD62p ,CD63 )and thrombopoietin( TPO)in preterm infants were recorded and compared. Results In chorioamnionitis group,the infant birth weight[(1. 90 ± 0. 41)kg vs. (2. 31 ± 0. 62)kg]and the PLT in 72 hours[<24 h(197. 97 ± 63. 43)×109/L vs.(266. 34 ± 69. 92)×109/L;24-72 h(202. 28 ± 29. 70)×109/L vs.(256. 38 ± 69. 96)×109/L]were significantly lower compared with the con-trol group,and the differences were statistically significant( all P <0. 05). The incidence of early thrombocytopenia (37. 50% vs. 9. 38% ),intracranial hemorrhage(40. 62% vs. 15. 63% ),MPV[(8. 73 ± 0. 89)fL vs.(8. 27 ± 0. 64)fL] and PDW[(59. 46 ± 5. 90)% vs.(55. 20 ± 5. 37)% ]in 24 hours were significantly higher in chorioamnionitis group, and the differences were statistically significant(all P<0. 05). The placental MVD in chorioamnionitis group signifi-cantly decreased[(9. 08 ± 1. 35)% vs.(12. 89 ± 1. 36)% ,P<0. 05 ]. The level of CD62p ,CD63 and TPO in umbilical cord blood were significantly higher in chorioamnionitis group[(25. 37 ± 5. 20)% vs.(10. 35 ± 2. 94)% ,(9. 49 ± 1. 58)% vs.(4. 04 ± 1. 21)% ,(271. 08 ± 197. 22)μg/L vs.(141. 87 ± 78. 10)μg/L,all P<0. 05]. The placental MVD was positively associated with PLT( r=0. 74,P<0. 05)and negatively associated with CD62p ,CD63 and TPO among infants with chorioamnionitis(r= -0. 64,-0. 44,-0. 44,all P<0. 05). Conclusions The chorioamnionitis may decrease the MVD in placenta and activate platelet in fetal circulation,damaged placental microvessel may activate platelet further.

Congenital tuberculosis after in vitro fertilization: suggestion for tuberculosis tests in infertile women in developing countries.

Congenital tuberculosis (CTB) after in vitro fertilization (IVF) is a major new problem in developing countries. Only 16 cases of CTB after IVF have been reported, and no tuberculosis (TB) tests were performed before IVF in these cases. However, on the basis of data in the literature and from the World Health Organization, the incidence of CTB has been substantially underestimated. We describe two cases of CTB after IVF in detail in our center and provide new insight into the important issue of controlling TB vertical transmission in developing countries. Performing an early diagnosis of CTB, mostly based on evidence of maternal infection and a high index of clinical suspicion, is a challenge. However, most mothers have no symptoms of TB infection during prepartum, and infertility may be the only symptom. Infertility caused by genital TB is common in countries with a high TB burden, and IVF is considered to be an effective treatment to improve their fertility. Therefore, this may lead to more CTB cases without thorough TB tests before IVF. We suggest that thorough TB tests should be conducted in infertile women before IVF to prevent CTB.

Predictive effect of neonatal morbidities on the poor outcomes at 12 months corrected age in very low birth weight premature infants / 中华儿科杂志

Objective@#To investigate the prognostic effect of neonatal morbidities on poor outcomes at 12 months corrected age in very low birth weight (VLBW) premature infants .@*Method@#From November 2013 to October 2014, a multi-center retrospective study was conducted in 8 tertiary Maternal and Children′s hospitals in Guangdong, Hunan and Fujian. The premature infants survived to a postmenstrual age (PMA) of 36 weeks with birth weight less than 1 500 g and without congenital diseases were included, and divided into two groups according to poor outcomes. The birth weight, gestational age, morbidities and poor outcomes (death, cerebral palsy, cognitive delay, et al) were recorded. Data were analyzed with Chi-square test to investigate the relationship between morbidities and poor outcomes. And the predictive effect of the top three morbidities were analyzed by Logistic regression analysis.@*Result@#Total of 834 VLBW premature infants (473 boys and 361 girls) finished the follow-up, whose average gestational age and birth weight were (30.6±1.8) weeks and (1 189±159)g. The incidences of BPD, severe ROP, NEC, brain injury and sepsis were 207 (24.8%), 119 (14.3%), 58 (7.0%), 281 (33.7%) and 124 (14.9%), respectively. There were significant differences between the two groups in the incidences of BPD, severe ROP, NEC, brain injury and sepsis(χ2=42.10, 47.20, 4.81, 44.28, 18.63, all P<0.01), which had significant correlation with poor outcomes at 12 months corrected age. The three top morbidities were severe ROP, BPD and brain injury(OR=3.82, 2.90, 2.80). Combined morbidities with BPD, severe ROP and brain injury correlated with higher risk of poor outcomes (one morbidity, OR=3.14, β=1.15; two morbidities, OR=7.31, β=1.99; three morbidities, OR=22.41, β=3.11; all P<0.01).@*Conclusion@#BPD, severe ROP, NEC, brain injury and sepsis were the risk factors of poor outcomes at 12 months corrected age in VLBW infants. And the more combined morbidities with severe ROP, BPD and brain injury, the higher risk of poor outcomes in this population. Trial registration Clinical Trails, NCT03104946.