Targeting TLR2/Rac1/cdc42/JNK Pathway to Reveal That Ruxolitinib Promotes Thrombocytopoiesis.

BACKGROUND: Thrombocytopenia has long been considered an important complication of chemotherapy and radiotherapy, which severely limits the effectiveness of cancer treatment and the overall survival of patients. However, clinical treatment options are extremely limited so far. Ruxolitinib is a potential candidate. METHODS: The impact of ruxolitinib on the differentiation and maturation of K562 and Meg-01 cells megakaryocytes (MKs) was examined by flow cytometry, Giemsa and Phalloidin staining. A mouse model of radiation-injured thrombocytopenia (RIT) was employed to evaluate the action of ruxolitinib on thrombocytopoiesis. Network pharmacology, molecular docking, drug affinity responsive target stability assay (DARTS), RNA sequencing, protein blotting and immunofluorescence analysis were applied to explore the targets and mechanisms of action of ruxolitinib. RESULTS: Ruxolitinib can stimulate MK differentiation and maturation in a dose-dependent manner and accelerates recovery of MKs and thrombocytopoiesis in RIT mice. Biological targeting analysis showed that ruxolitinib binds directly to Toll Like Receptor 2 (TLR2) to activate Rac1/cdc42/JNK, and this action was shown to be blocked by C29, a specific inhibitor of TLR2. CONCLUSIONS: Ruxolitinib was first identified to facilitate MK differentiation and thrombocytopoiesis, which may alleviate RIT. The potential mechanism of ruxolitinib was to promote MK differentiation via activating the Rac1/cdc42/JNK pathway through binding to TLR2.

A Novel Antithrombocytopenia Agent, Rhizoma cibotii, Promotes Megakaryopoiesis and Thrombopoiesis through the PI3K/AKT, MEK/ERK, and JAK2/STAT3 Signaling Pathways.

BACKGROUND: Cibotii rhizoma (CR) is a famous traditional Chinese medicine (TCM) used to treat bleeding, rheumatism, lumbago, etc. However, its therapeutic effects and mechanism against thrombocytopenia are still unknown so far. In the study, we investigated the effects of aqueous extracts of Cibotii rhizoma (AECRs) against thrombocytopenia and its molecular mechanism. METHODS: Giemsa staining, phalloidin staining, and flow cytometry were performed to measure the effect of AECRs on the megakaryocyte differentiation in K562 and Meg-01 cells. A radiation-induced thrombocytopenia mouse model was constructed to assess the therapeutic actions of AECRs on thrombocytopenia. Network pharmacology and experimental verification were carried out to clarify its mechanism against thrombocytopenia. RESULTS: AECRs promoted megakaryocyte differentiation in K562 and Meg-01 cells and accelerated platelet recovery and megakaryopoiesis with no systemic toxicity in radiation-induced thrombocytopenia mice. The PI3K/AKT, MEK/ERK, and JAK2/STAT3 signaling pathways contributed to AECR-induced megakaryocyte differentiation. The suppression of the above signaling pathways by their inhibitors blocked AERC-induced megakaryocyte differentiation. CONCLUSIONS: AECRs can promote megakaryopoiesis and thrombopoiesis through activating PI3K/AKT, MEK/ERK, and JAK2/STAT3 signaling pathways, which has the potential to treat radiation-induced thrombocytopenia in the clinic.

Associations of Maternal Fructosamine before Delivery in Gestational Diabetes Mellitus Pregnancies with Neonatal Glucometabolic Disorders.

Background: The offspring of pregnant women with gestational diabetes mellitus (GDM) are vulnerable to be glucometabolic disorders. However, to date, few current studies focused on the associations of maternal accumulated glucose exposure before delivery with neonatal glucometabolic disorders and large for gestational age (LGA) infants. This study is aimed at exploring the associations of maternal fructosamine (FMN) before delivery in GDM pregnant women with neonatal glucometabolic disorders in the first 3 days of life and LGA infants. Methods: The study subjects were the GDM pregnant women, who gave birth in our hospital from September 1, 2018 to January 31, 2021, and their newborns. The maternal FMN adjusted by serum albumin (FMNALB) before delivery was selected as exposure factors. A multivariate logistical regression model was used to calculate the odds ratios (OR) for neonatal glucometabolic disorders, hypoglycemia needing intervention (<2.6 mmol/L), and glucose intolerance (>7.0 mmol/L) in the first 3 days and LGA infants. Results: In GDM pregnant women, the newborns in the maternal FMNALB ≥ 75th percentile (≥5.89 mmol/g) group had higher risks in neonatal glucometabolic disorders (aOR 2.50, 95% CI 1.34-4.65, P = 0.004) and hypoglycemia (aOR 2.18, 95% CI 1.16-4.10, P = 0.016). However, FMNALB ≥ 75th percentile seemed to be not predictive of the glucose intolerance (aOR 1.76, 95% CI 0.82-3.79, P = 0.149) and LGA (aOR 1.56, 95% CI 0.81-3.02, P = 0.185). Further, in the sensitivity analysis, the newborns in the maternal FMNALB ≥ 90th percentile (≥6.40 mmol/g) group also had higher risks in neonatal glucometabolic disorders (aOR 5.70, 95% CI 2.18-14.89, P < 0.001) and hypoglycemia (aOR 3.72, 95% CI 1.48-9.31, P = 0.005). Conclusions: The maternal FMNALB before delivery in GDM pregnant women was a useful biomarker to identify the offspring with high risk of neonatal glucometabolic disorders. However, the association between maternal FMNALB and the risk of LGA infants was not so strong.

Associations between Neonatal Serum Bilirubin and Childhood Obesity in Term Infants.

Inverse correlations between serum bilirubin level and obesity had been reported in adults. We aimed to investigate the associations between neonatal hyperbilirubinemia and childhood obesity. Data was obtained from the U.S. Collaborative Perinatal Project (CPP), a multicenter study from 1959 to 1976. Data of serum bilirubin in term newborns were used to observe the association with obesity at age of 7 years. Logistic regression models were performed to calculate adjusted odds ratios (aORs) for obesity. For children from the same mother sharing similar factors, Generalized Estimating Equation (GEE) model was used to correct for intracluster correlation. Relative to newborns with total serum bilirubin (TSB) < 3 mg/dl, there are lower risks for obesity in those with 3 mg/dl ≤ TSB < 6 mg/dl (aOR 0.91; 95%CI 0.81, 1.02), 6 mg/dl ≤ TSB < 9 mg/dl (aOR 0.88; 95%CI 0.78, 0.99), 9 mg/dl ≤ TSB<13 mg/dl (aOR 0.83; 95%CI 0.71, 0.98). By stratifying for subtypes of bilirubin, the inverse correlations only existed in exposure to unconjugated bilirubin. By using the GEE model correcting for intracluster correlations, the results are consistent. In summary, exposure to bilirubin up to 13 mg/dl is inversely associated with obesity at the age of 7 years in term infants.

Associations between neonatal serum bilirubin and childhood hypertension.

Mild hyperbilirubinemia is inversely associated with cardiometabolic diseases in adults. The aim of this study was to evaluate the association between neonatal serum bilirubin levels and childhood hypertension. Data were obtained from the U.S. Collaborative Perinatal Project conducted at 12 U.S. medical centers from 1959 to 1965. This multicenter study recruited participants before phototherapy was routinely used, thereby excluding the influence of phototherapy. In 37,544 newborns (31,819 term and 5,725 preterm births), a generalized linear model and a logistic regression model were used to calculate the linear coefficients and adjusted odds ratios (ORs) of blood pressure and hypertension at 7 years of age based on neonatal serum bilirubin levels. No significant correlation was observed between serum bilirubin at 48 hours after birth and blood pressure at the age of 7 years in the whole study population and in the subgroup of term infants. In preterm infants, a lower total serum bilirubin and unconjugated bilirubin of 3 mg/dl were associated with a higher systolic blood pressure of 62 mmHg (0.38-0.86, p <0.001) and 0.70 mmHg (0.10-1.30, p <0.05) respectively. Relative to a total serum bilirubin level <3 mg/dl among preterm infants, total serum bilirubin levels of 3-6 mg/dl (adjusted OR 1.36; 95% CI: 0.98-1.89), 6-9 mg/dl (adjusted OR 1.35; 95% CI: 0.98-1.85), 9-12 mg/dl (adjusted OR 1.55; 95% CI: 1.10-2.19), and ≥12 mg/dl (adjusted OR 1.42; 95% CI: 1.01-2.00) were associated with higher risks of hypertension. After stratifying for the subtypes of bilirubin, the associations only existed for unconjugated bilirubin. In addition, consistent findings existed when using maximum neonatal serum bilirubin as an exposure factor. Neonatal serum bilirubin levels are positively associated with childhood blood pressure/hypertension in preterm infants. Our findings may shed some light on the role of bilirubin in the prevention of hypertension.

Association between neonatal serum bilirubin and childhood obesity in preterm infants.

BACKGROUND: Serum bilirubin levels are inversely associated with obesity in adults. We are interested in whether neonatal jaundice is associated with childhood obesity in preterm infants. METHODS: Data were obtained from the US Collaborative Perinatal Project. Neonatal bilirubin levels were used as exposure factors for obesity at age 7 years. Logistic regression models were used to control for potential confounders and calculate odds ratios (ORs). A generalized estimating equation (GEE) model was used to correct for intracluster correlation coefficient. SAS was used for all statistical analyses. RESULTS: In the study subjects, 865 of 5019 preterm infants were obese at age 7 years. While neonatal total serum bilirubin (TSB) rose 1 mg/dl, body mass index (BMI) increased 0.03 kg/m2 (95% confidence interval (CI) 0.02, 0.04). Compared with infants with TSB <3 mg/dl, the ORs (95% CIs) for obesity in infants with 3 mg/dl≤ TSB <6 mg/dl, 6 mg/dl≤ TSB <9 mg/dl, 9 mg/dl≤ TSB <12 mg/dl and TSB ≥12 mg/dl were, respectively, 1.18 (0.87, 1.59), 1.25 (0.93, 1.67), 1.52 (1.11, 2.09), and 1.67 (1.22, 2.07). By using subtypes of bilirubin as exposure factors and the GEE model to correct for intracluster correlation coefficient, similar trends of associations were observed. CONCLUSION: Neonatal bilirubin levels have positive trends of associations with childhood obesity in preterm infants.

Gender Differences in Infant Mortality and Neonatal Morbidity in Mixed-Gender Twins.

In the present study, we aimed to explore gender differences in infant mortality and neonatal morbidity in mixed-gender twin pairs. Data were obtained from the US National Center for Health Statistics Linked Birth-Infant Death Cohort. A total of 108,038 pairs of mixed-gender twins were included in this analysis. Among the mixed-gender twins, no significant difference in the odds of fetal mortality between male twins (1.05%) and female co-twins (1.04%). However, male twins were at increased odds of neonatal mortality (adjusted OR 1.59; 95% CI 1.37, 1.85) and overall infant mortality (adjusted OR 1.43; 95% CI 1.27, 1.61) relative to their female co-twins. Congenital abnormalities (adjusted OR 1.38; 95% CI 1.27, 1.50) were identified significantly more frequently in male than female twins. Moreover, increased odds of having low 5-minute Apgar score (<7) (adjusted OR 1.15; 95% CI 1.05, 1.26), assistant ventilation >30 minutes (adjusted OR 1.31; 95% CI 1.17, 1.47), and respiratory distress syndrome (adjusted OR 1.45; 95% CI 1.26, 1.66) were identified in male twins relative to their female counterparts. The results of our study indicated that in mixed-gender twin pairs, the odds of infant mortality and neonatal morbidity were higher in male twins than their female co-twins.

Creating a placental inflammatory composite index that has a high prognostic relevance to child morbidity.

AIM: Selecting pathologic measures of placental inflammation that affect pregnancy and childhood health is largely empirical. We aimed to systematically select several core inflammation-related placental measures to construct a novel placental inflammatory evaluation criterion with a high prognostic relevance to child morbidity. METHODS: We used data from the US Collaborative Perinatal Project (1959-1976), a longitudinal birth cohort study that recruited women during pregnancy and followed the children until 7 years of age. Bootstrap resampling, least absolute shrinkage and selection operator, and receiver-operator curve were used to select placental pathologic measures that were closely related to child morbidity to form a placental inflammatory composite index. RESULTS: Twenty-six candidate placental inflammation-related measures were ranked based on their close association with adverse neonatal outcomes. The top five placental measures were: (i) neutrophilic infiltration in umbilical artery; (ii) placental weight-birthweight ratio; (iii) necrosis in decidua capsularis; (iv) bacterial colony in epithelium of amnion; and (v) opacity of membranes and fetal surface. Several composite indexes were constructed. A five-measure composite index that had the highest prognostic relevance was chosen. Compared with subjects without any of the five abnormal measures, those with any lesion ranging from 1 to 5 had a 1.2- to 4.6-fold risk of adverse child outcomes, respectively. CONCLUSION: Our composite index is simple, evidence-based, and has predictive value for child morbidity. It may be used as a novel placental inflammatory evaluation criterion.