Correlation between receptor-interacting protein 140 expression and directed differentiation of human embryonic stem cells into neural stem cells

Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.

Overexpression or knock-down of runt-related transcription factor 1 affects BCR-ABL-induced proliferation and migration in vitro and leukemogenesis in vivo in mice

Background Runt-related transcription factor 1 （Runxl） plays a crucial role in hematogenesis and its dysfunction may contribute to leukemogenesis. However, it is not clear whether or not abnormal expression of Runxl will induce leukemia and how the change of Runxl expression level could affect BCR-ABL-induced leukemogenesis. In the present study, we aimed to analyze if abnormal expression of Runxl in BaF3 cells alone would induce teukemogenesis. And we also wanted to know if abnormal expression of Runxl in leukemic cells would affect leukemogenesis. Furthermore, we investigated whether overexpression or knock-down of Runxl in BaF3 cells would induce leukemogenesis. Methods Plasmids containing full-length Runxl cDNA were transduced into BaF3 cells and BaF3-P185wt cells （BCR-ABL transformed BaF3 cells） by electroporation. Plasmids containing a short hairpin RNA of Runxl were transduced into BaF3 cells and BaF3-P185wt cells by electroporation. Runxl expression level was quantified by Western blotting and quantitative real-time PCR. The effects of overexpression or knock-down of Runxl on proliferation, apoptosis and migration of cells were detected in vitro. Then, using MSCV-P185wt-EGFP as a control, we transplanted MSCV-P185wt-Runx1 cells or MSCV-P185wt-shRNA cells into Balb/c mice through tail vein and observed tumorgenesis of the different phenotypes. Results In vitro analysis revealed that overexpression of Runxl in P185wt cells could inhibit cell proliferation and slow down cell migration; while knock-down of Runxl could promote cell proliferation and speed up cell migration. In vivo analysis indicated that mice transplanted with MSCV-P185wt-Runx1 survived longer than controls. In contrast, mice transplanted with MSCV-P185wt-shRNA survived shorter than the control group. Gross pathological analysis revealed that the MSCV-P185wt-Runx1 group had less severe splenomegaly and hepatomegaly compared to the control group, and the MSCV-P185wt-shRNA group had more severe splenomegaly and hepatomegaly. No splenomegaly or hepatomegaly was detected in mice transplanted with MSCV-BaF3-Runxl cells or MSCV-BaF3-shRNA cells. Both the mice of MSCV-BaF3-Runxl group and MSCV-BaF3-shRNA group were healthy with no sign of leukemia for up to three months. Conclusions Overexpression or knock-down of Runxl gene in BaF3 cells alone could not induce leukemogenesis. However, in BaF3-P185wt cells, alteration of Runxl expression could affect BCR-ABL-induced proliferation and migration in vitro and leukemoaenesis in vivo.

Highly efficient gene editing and single cell analysis of hematopoietic stem/progenitor cells from X-linked sideroblastic anemia patients

Dear Editor,X-linked sideroblastic anemia(XLSA),which is the most common genetic form of congenital sideroblastic anemia,is typically characterized by reduced heme synthesis and the presence of bone marrow(BM)ring sideroblasts containing pathologic iron deposits in the mitochondria.

Delivery room resuscitation and short-term outcomes of extremely preterm and extremely low birth weight infants: a multicenter survey in North China

Background:Delivery room resuscitation assists preterm infants,especially extremely preterm infants(EPI)and extremely low birth weight infants(ELBWI),in breathing support,while it potentially exerts a negative impact on the lungs and outcomes of preterm infants.This study aimed to assess delivery room resuscitation and discharge outcomes of EPI and ELBWI in China.Methods:The clinical data of EPI(gestational age[GA]<28 weeks)and ELBWI(birth weight[BW]<1000 g),admitted within 72 h of birth in 33 neonatal intensive care units from five provinces and cities in North China between 2017 and 2018,were analyzed.The primary outcomes were delivery room resuscitation and risk factors for delivery room intubation(DRI).The secondary outcomes were survival rates,incidence of bronchopulmonary dysplasia(BPD),and risk factors for BPD.Results:A cohort of 952 preterm infants were enrolled.The incidence of DRI,chest compressions,and administration of epinephrine was 55.9%(532/952),12.5%(119/952),and 7.0%(67/952),respectively.Multivariate analysis revealed that the risk factors for DRI were GA<28 weeks(odds ratio[OR],3.147;95%confidence interval[CI],2.082–4.755),BW<1000 g(OR,2.240;95%CI,1.606–3.125),and antepartum infection(OR,1.429;95%CI,1.044–1.956).The survival rate was 65.9%(627/952)and was dependent on GA.The rate of BPD was 29.3%(181/627).Multivariate analysis showed that the risk factors for BPD were male(OR,1.603;95% CI,1.061–2.424),DRI(OR,2.094;95% CI,1.328–3.303),respiratory distress syndrome exposed to≥2 doses of pulmonary surfactants(PS;OR,2.700;95%CI,1.679–4.343),and mechanical ventilation≥7 days(OR,4.358;95% CI,2.777–6.837).However,a larger BW(OR,0.998;95% CI,0.996–0.999),antenatal steroid(OR,0.577;95%CI,0.379–0.880),and PS use in the delivery room(OR,0.273;95%CI,0.160–0.467)were preventive factors for BPD(all P<0.05).Conclusion:Improving delivery room resuscitation and management of respiratory complications are imperative during early management of the health of EPI and ELBWI.

Receptor-interacting Protein 140 Overexpression Promotes Neuro-2a Neuronal Differentiation by ERK1/2 Signaling

Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of genes in trisomic cells.Receptor-interacting protein 140 （RIP 140） is significantly upregulated in DS brains,suggesting its involvement in DS CNS development abnormalities.However,the role of RIP140 in neuronal differentiation is still not clear.The current study aimed to investigate the effect of RIP 140 overexpression on the differentiation of neuro-2a （N2a） neuroblastoma cells,in vitro.Methods：Stably RIP 140-overexpressing N2a （N2a-RIP140） cells were used as a neurodevelopmental model,and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot.Retinoic acid （RA） was used to stimulate N2a differentiation.Combining the expression of Tuj 1 at the mRNA and protein levels,the percentage of cells baring neurites,and the number of neurites per cell body was semi-quantified to determine the effect of RIP 140 on differentiation of N2a cells.Furthermore,western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP 140 induces differentiation of N2a cells.Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.Results：Compared to untransfected N2a cells RIP140 expression in N2a-RIP 140 cells was remarkably upregulated at both the mRNA and protein levels.N2a-RIP 140 cells had a significantly increased percentage of cells baring neurites,and numbers of neurites per cell,as compared to N2a cells,in the absence and presence of RA （P 〈 0.05）.In addition,Tuj l,a neuronal biomarker,was strongly upregulated in N2a-RIP140 cells （P 〈 0.05） and phosphorylated ERK1/2 （p-ERK1/2） levels in N2a-RIP140 cells were dramatically increased,while differentiation was inhibited by the ERK 1/2-specific inhibitor U0126.Conclusions：RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.

Oxcarbazepine oral suspension in pediatric patients with partial seizures and/or generalized tonic-clonic seizures: a multi-center, single arm, observational study in China

Background:To assess efficacy and safety of oxcarbazepine (OXC) oral suspension in pediatric patients aged 2-16 years with partial seizures (PS) and/or generalized tonic-clonic seizures (GTCS) in real-world clinical practice in China.Methods:This 26-week,single arm,multicenter and observational study recruited patients aged 2-16 years with PS or GTCS suitable for OXC oral suspension treatment.Enrolled patients received OXC oral suspension treatment for 26 weeks.Primary endpoints included mean seizure frequency at the end of the treatment and mean seizure frequency reduction at the end of the treatment vs.baseline.Secondary efficacy-related endpoints and safety parameters were also assessed.Results:Nine hundred and eighty-seven pediatric patients were enrolled and 912 (92.4%) completed the study.The mean seizure frequencies at baseline and the end of week 26 were 13.40±64.92 and 1.62±19.47 times/month,respectively.The mean seizure frequency reduction was 10.03±63.67 times/month and the mean seizure frequency reduction percentage was 90.02%±5127.0% (P<0.0001).After 26 weeks of treatment,82.36%,7.24% and 3.86% of the patients became controlled,significantly improved and improved,respectively.Adverse events (AEs) were reported in 74 (7.65%) patients.Rash was the most common AE.The efficacy of OXC was not affected by seizure types,age or gender.Conclusion:This study confirms the efficacy and good safety profile of OXC oral suspension in Chinese pediatric patients aged 2-16 years with PS and/or GTCS.

Haploidentical hematopoietic stem cell transplantation may improve long-term survival for children with high-risk T-cell acute lymphoblastic leukemia in first complete remission

Background: The role of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with high-risk (HR) T-cell acute lymphoblastic leukemia (T-ALL) in first complete remission (CR1) is still under evaluation. Moreover, relapse is the main factor affecting survival. This study aimed to explore the effect of allo-HSCT (especially haploidentical HSCT [haplo-HSCT]) on improving survival and reducing relapse for HR childhood T-ALL in CR1 and the prognostic factors of childhood T-ALL in order to identify who could benefit from HSCT.Methods: A total of 74 newly diagnosed pediatric T-ALL patients between January 1, 2012 and June 30, 2018 were enrolled in this retrospective study. Patients were stratified into the low-risk chemotherapy cohort (n = 16), HR chemotherapy cohort (n = 31), and HR transplant cohort (n = 27). Characteristics, survival outcomes, and prognostic factors of all patients were then analyzed.Results: Patient prognosis in the HR chemotherapy cohort was significantly worse than that in the low-risk chemotherapy cohort (5-year overall survival [OS]: 58.5%vs. 100%,P = 0.003;5-year event-free survival [EFS]: 54.1%vs. 83.4%,P = 0.010;5-year cumulative incidence of relapse [CIR]: 45.2%vs. 6.3%,P = 0.011). In HR patients, allo-HSCT improved the 5-year EFS and CIR compared to that of chemotherapy (5-year EFS: 80.1%vs. 54.1%,P = 0.041;5-year CIR: 11.6%vs. 45.2%,P = 0.006). The 5-year OS was higher in the HR transplant cohort than that in the HR chemotherapy cohort (81.0%vs. 58.5%,P = 0.084). Minimal residual disease re-emergence was an independent risk factor for 5-year OS, EFS, and CIR;age ≥10 years was an independent risk factor for OS and EFS;and high white blood cell count was an independent risk factor for EFS and CIR.Conclusion: Allo-HSCT, especially haplo-HSCT, could effectively reduce relapse of children with HR T-ALL in CR1.

Perinatal risk factors and neonatal complications in discordant twins admitted to the neonatal intensive care unit

Background Many studies have shown a relationship between birth weight discordance and adverse perinatal outcomes. This study aimed to investigate the perinatal risk factors and neonatal complications of discordant twins who are admitted to the neonatal intensive care unit. Methods A total of 87 sets of twins were enrolled in this retrospective study, of which 22 sets were discordant twins and 65 sets were concordant twins. Binary Logistic regression analysis was used to identify the risk factors associated with the occurrence of discordant twins. The common neonatal complications of discordant twins were also investigated. Results Multivariate analysis showed that the use of assisted reproductive techniques, pregnancy-induced hypertension, and unequal placental sharing were risk factors for the occurrence of discordant twins. The incidence of small for gestational age infants and very low birth weight infants of discordant twins was significantly higher, while the birth weight of discordant twins was significantly lower than those of concordant twins. The duration of hospitalization of discordant twins was longer than that of concordant twins. The incidence of several neonatal complications, such as neonatal respiratory distress syndrome and intracranial hemorrhage, was higher in discordant twins than that in concordant twins. The percentage of those requiring pulmonary surfactant and mechanical ventilation was significantly higher in discordant twins than that in concordant twins. Conclusions Use of assisted reproductive techniques, pregnancy-induced hypertension, and unequal placental sharing are perinatal risk factors of discordant twins who are admitted to the neonatal intensive care unit. These infants are also much more likely to suffer from various neonatal complications, especially respiratory and central nervous system diseases. It is important to prevent the occurrence of discordant twins by decreasing these risk factors and timely treatment should be given to discordant twins.

Chinese expert recommendations on ketogenic diet therapy for super-refractory status epilepticus

Super-refractory status epilepticus(SRSE)is a serious and life-threatening neurological condition.Ketogenic diet(KD)is a diet characterized by high fat,low carbohydrate,and moderate protein.As KD shows effectiveness in controlling seizures in more than half of SRSE patients,it can be a treatment option for SRSE.Currently,KD treatment for SRSE is based on personal experience and observational evidence has been published.In the context of a lack of a validated guideline,we convened a multicenter expert panel within the China Association Against Epilepsy(CAAE)Ketogenic Diet Commission to work out the Chinese expert recommendations on KD for SRSE.We summarize and discuss the latest clinical practice of KD for SRSE in critical care settings.Recommendations are given on patient selection,the timing of KD,diet implementation,and follow-up.More research data are needed in this area to support better clinical practice.