Unrelated umbilical cord blood transplantation with low-dose anti-thymocyte globulin for children with severe aplastic anemia: A case series.

OBJECTIVE: This study aimed to investigate the prognosis of unrelated umbilical cord blood transplantation (UCBT) using low-dose anti-thymocyte globulin (ATG) in children diagnosed with severe aplastic anemia (SAA). METHODS: This retrospective case series study was conducted involving pediatric SAA patients treated at the Capital Institute of Pediatrics from January 2020 to February 2023. All patients underwent a reduced-intensity conditioning (RIC) regimen alongside low-dose ATG. RESULTS: The study comprised nine patients (five males) with a median age of 5 years (range: 1.7 to 7 years). The median follow-up duration was 799 days (range: 367 to 1481 days), during which all patients survived. The median time interval from diagnosis to transplantation was 3 months (range: 1 to 9 months). The median dosage of ATG administered was 5 mg/kg (range: 2.5 to 7.5 mg/kg). The median durations for granulocyte and platelet engraftment were 15 days (range: 12 to 23 days) and 26 days (range: 12 to 41 days), respectively. Three patients experienced grade 2-4 acute graft-versus-host disease (aGVHD). Epstein-Barr virus (EBV) reactivation was observed in three patients, while cytomegalovirus (CMV) reactivation occurred in seven patients, with no cases of CMV disease or post-transplant lymphoproliferative disorder (PTLD). One patient experienced recurrence 15 months after transplantation due to influenza A infection. CONCLUSION: These findings indicate that SAA patients may attain a favorable prognosis following UCBT with a RIC regimen combined with low-dose ATG.

Human behavior in free search online shopping scenarios can be predicted from EEG activation using Hjorth parameters.

The present work investigates whether and how decisions in real-world online shopping scenarios can be predicted based on brain activation. Potential customers were asked to search through product pages on e-commerce platforms and decide, which products to buy, while their EEG signal was recorded. Machine learning algorithms were then trained to distinguish between EEG activation when viewing products that are later bought or put into the shopping card as opposed to products that are later discarded. We find that Hjorth parameters extracted from the raw EEG can be used to predict purchase choices to a high level of accuracy. Above-chance predictions based on Hjorth parameters are achieved via different standard machine learning methods with random forest models showing the best performance of above 80% prediction accuracy in both 2-class (bought or put into card vs. not bought) and 3-class (bought vs. put into card vs. not bought) classification. While conventional EEG signal analysis commonly employs frequency domain features such as alpha or theta power and phase, Hjorth parameters use time domain signals, which can be calculated rapidly with little computational cost. Given the presented evidence that Hjorth parameters are suitable for the prediction of complex behaviors, their potential and remaining challenges for implementation in real-time applications are discussed.

Clinical features and prognosis of pediatric acute lymphocytic leukemia with JAK-STAT pathway genetic abnormalities: a case series.

The objective of this study is to explore the clinical features and outcomes of pediatric patients with acute lymphoblastic leukemia (ALL) harboring JAK-STAT signaling pathway genetic abnormalities. This retrospective case series examined the clinical data of pediatric patients diagnosed with ALL harboring JAK-STAT pathway genetic abnormality at the Children's Hospital of the Capital Institute of Pediatrics between January 2016 and January 2022. Bone marrow next-generation sequencing was used to reveal the JAK pathway abnormalities. Descriptive statistics were used. From 432 children with ALL during the study period, eight had JAK-STAT pathway genetic abnormalities. Regarding immunotyping, there were four patients with common-B cell types and one with pre-B cell type. The three patients with T-ALL had early T-cell precursor(ETP) type, pre-T cell type, and T cell type. Gene mutations were more common than fusion genes. There was no central nervous system involvement in eight patients. All patients were considered at least at intermediate risk before treatments. Four patients underwent hematopoietic stem cell transplantation (HSCT). One child had a comprehensive relapse and died. The child had a severe infection and could not tolerate high-intensity chemotherapy. Another child relapsed 2 years after HSCT and died. Disease-free survival was achieved in six children. JAK-STAT pathway genetic abnormalities in pediatric Ph-like ALL are rare. Special attention should be paid to treatment-related complications, such as infection and combination therapy (chemotherapy, small molecule targeted drugs, immunotherapy, etc.) to reduce treatment-related death and improve long-term quality of life.

Clinical outcomes of individualized busulfan-dosing in hematopoietic stem cell transplantation in Chinese children undergoing with therapeutic drug monitoring.

To identify relationships between busulfan (Bu) exposure and outcomes of a cohort pediatric patients receiving hematopoietic stem cell transplantation (HSCT), along with a targeted busulfan-based conditioning regimen. We retrospectively evaluated targeted busulfan concentrations in 53 pediatric patients (age 0.4-16 years) who received busulfan 4 times daily according to recommended weight-based doses in a single-center analysis between 2018 and 2020. In this trial, individual busulfan pharmacokinetics were performed following dose 5 of the conditioning regimen. Twenty four of 53 patients (45.3%) studies did not require dose adjustments. Equal number of patients (24/53) required one dose adjustments while two-dose adjustment applied for 5 of 53 (9.4%). Twenty-one percent of the patients exhibited ll-lV aGVHD. The incidence of veno-occlusive disease (VOD) was in 3.8% of the 53 patients, while incidence of hemorrhagic cystitis (II-III) reached to 9.7%. Engraftment was successful in 98% of the 53 patients with relapse in 2% of cases. The probability of overall survival and disease-free survival at day 100 was 96% and 94%, respectively. In conclusion, therapeutic drug monitoring (TDM) and individualization of Bu dosage are essential to improve the efficacy and safety of busulfan-based regimen in Chinese pediatric HSCT recipients.

Neural Signature of Buying Decisions in Real-World Online Shopping Scenarios - An Exploratory Electroencephalography Study Series.

The neural underpinnings of decision-making are critical to understanding and predicting human behavior. However, findings from decision neuroscience are limited in their practical applicability due to the gap between experimental decision-making paradigms and real-world choices. The present manuscript investigates the neural markers of buying decisions in a fully natural purchase setting: participants are asked to use their favorite online shopping applications to buy common goods they are currently in need of. Their electroencephalography (EEG) is recorded while they view the product page for each item. EEG responses to pages for products that are eventually bought are compared to those that are discarded. Study 1 repeats this procedure in three batches with different participants, product types, and time periods. In an explorative analysis, two neural markers for buying compared to no-buying decisions are discovered over all three batches: frontal alpha asymmetry peak and frontal theta power peak. Occipital alpha power at alpha asymmetry peaks differs in only one of the three batches. No further significant markers are found. Study 2 compares the natural product search to a design in which subjects are told which product pages to view. In both settings, the frontal alpha asymmetry peak is increased for buying decisions. Frontal theta peak increase is replicated only when subjects search through product pages by themselves. The present study series represents an attempt to find neural markers of real-world decisions in a fully natural environment and explore how those markers can change due to small adjustments for the sake of experimental control. Limitations and practical applicability of the real-world approach to studying decision-making are discussed.

A correlation analysis of miRNA­34a and its predicted target genes in leukemia.

microRNA­34a (miRNA­34a) plays an important role in the pathogenesis of leukemia. This study aimed to explore its role in the proliferation of HL­60 cells and the correlation with some of its predicted target genes: the cyclin­dependent kinase 4 (CDK4), the oncogene MYB and the silent information regulator 1 (SIRT1). We first analyzed the expression of miR­34a, CDK4, MYB and SIRT1 in peripheral blood samples from acute leukemia (AL) patients and healthy controls, and conducted a correlation analysis. HL­60 cells were then transfected with miR­34a and control 'scramble̓ miRNA, and quantitative RT­PCR and western blotting were used to analyze the effects of the interfering sequence in HL­60 cells. The expression of miR­34a was significantly reduced in AL patients compared to healthy controls (P<0.01), and negatively correlated with the expression of CDK4 and MYB. Sub­group analysis revealed that the expression of MYB was significantly lower in AL children <3 years old compared to those >3 years. Following the transfection of HL­60 cells with miR­34a, the mRNA level of CDK4, MYB and SIRT1 decreased by 53.2, 43.3 and 33.5%, respectively, compared to the control, similarly to the respective changes in protein levels. This study showed that the expression of miR­34a negatively correlates with the expression of CDK4 and MYB in pediatric patients with acute leukemia. miRNA­34a downregulates the expression of the CDK4, MYB and SIRT1 genes in vitro; it may thus represent a novel therapeutic target for acute leukemia.

Defective anchoring of JNK1 in the cytoplasm by MKK7 in Jurkat cells is associated with resistance to Fas-mediated apoptosis.

The c-Jun N-terminal protein kinase (JNK) plays a context-dependent role in tumorigenesis. Stress-induced redistribution of JNK from the cytoplasm to the nucleus has been demonstrated as essential for stress-induced cell death. However, accumulation of basal JNK activity in the nucleus has frequently been seen in tumor cells. Our previous report revealed aberrant nuclear entry of JNK protein in Jurkat human leukemic T-cells even without JNK hyperactivation. Because inhibition of JNK activity, especially JNK1 activity, in Jurkat cells results in augmented Fas-mediated apoptosis, it is possible that aberrant subcellular localization of JNK, especially the JNK1 isoform, contributes to the resistance to Fas-mediated apoptosis. Here we report that MKK7 works as a cytoplasmic anchoring protein for JNK1 in various types of cells, including human peripheral blood mononuclear cell (PBMC) T-cells, but exhibits aberrant nuclear entry in Jurkat cells. Ectopic expression of a JNK1 mutant defective of nuclear entry or a nuclear JNK inhibitor leads to impaired UV-induced apoptosis in both PBMC T- and Jurkat cells. The same treatment shows no effect on Fas-mediated apoptosis of PBMC T-cells but sensitizes Jurkat cells to Fas-mediated apoptosis. Taken together, our work suggests that aberrant subcellular organization of the JNK pathway might render certain tumor cells resistant to Fas-mediated apoptosis.

Silencing of the MYCN gene by siRNA delivered by folate receptor-targeted liposomes in LA-N-5 cells.

INTRODUCTION: MYCN amplification is highly associated with malignancy and correlates with poor prognosis in patients with neuroblastoma. MATERIALS AND METHODS: We developed a novel liposome-MYCN siRNA-folic acid complex, and the transfection efficacy was measured in LA-N-5 cells by cy-3 fluorescence density in each microgram of protein from the transfected cell lysate. MYCN expression and cell growth were studied with quantitative RT-PCR and MTT assays, and the expression of MYCN protein was studied with Western blot, respectively. An SCID mouse model with subcutaneous LA-N-5 xenografted tumor was established. The animals were divided into four groups (n = 5) and they were peritoneally injected with liposome-encapsulated MYCN siRNA (siRNA 125 µg/kg/day), lipid-encapsulated control siRNA, MYCN siRNA, or liposome only, respectively, for 5 consecutive days. The animals were killed 24 h after the last injection, and the expression of MYCN mRNA in tumor tissue was detected by RT-PCR. RESULTS: Our results are as follows: the transfect efficacy reached 1808.5 ± 140.2 pg siRNA/µg protein in LA-N-5 lysates after treatment with 100 nmol/L MYCN siRNA encapsulated with lipid, and fluorescence could be visualized in 92% of LA-N-5 cells after transfection. At 72 h post-transfection, MYCN mRNA expression in LA-N-5 cells was downregulated by 79.2%, MYCN protein was downregulated by 71.3% and cell growth was inhibited by 66.2%, as measured by MTT assay. In the in vivo study, MYCN mRNA expression was knocked down 53.1% in tumor tissues with injection of liposome-encapsulated MYCN siRNA as compared to control siRNA. CONCLUSION: These results suggest that targeted delivery of MYCN siRNA by folate receptor-targeted lipid vesicles into LA-N-5 cells is efficacious and capable of suppressing MYCN mRNA expression both in vitro and in vivo.

All-trans retinoic acid is capable of inducing folate receptor ß expression in KG-1 cells.

The high expression of folate receptor (FR) on cancer cells might be a potential target for cancer therapy. In this study, the FR-ß expression and the modulation effect of all-trans retinoic acid (ATRA) in a number of cancer cell lines were analyzed. The gateway of ATRA activity on FR-ß expression was further studied by a panel of retinoid activators and inhibitors. The results revealed that ATRA was capable of upregulating the expression of FR-ß protein in KG-1 cells in a dosage-dependent manner, not in KG-1a, NB4, HL60, 293, L1210, JAR, and Hela cells. FR-ß mRNA expression in KG-1 cells was higher when ATRA was present in culture medium at 10⁻6 mol/L for 5 days, and it went down to baseline when ATRA was removed from the medium, vice versa. The upregulation of FR-ß expression in KG-1 cells by ATRA was not associated with cell proliferation and differentiation. In addition, activators of retinoid acid receptor (RAR)α and RARγ, CD336, and CD2781 also induced FR-ß expression. The induction of FR-ß expression by CD336 could be inhibited by RARγ antagonist CD2665; RARß agonist CD-417 and CD-2314 as well as retinoid X receptor (RXR) agonist LG100364 could not induce FR-ß expression. These results indicate that ATRA within a certain range of concentration could reversibly induce the expression of FR-ß in a dosage- and cell type-dependent manner, and its action in KG-1 cells might be associated with the signal transduction of retinoid receptor RARα and RARγ, rather than RARß and RXRs.

Selective unresponsiveness to the inhibition of p38 MAPK activation by cAMP helps L929 fibroblastoma cells escape TNF-alpha-induced cell death.

BACKGROUND: The cyclic AMP (cAMP) signaling pathway has been reported to either promote or suppress cell death, in a cell context-dependent manner. Our previous study has shown that the induction of dynein light chain (DLC) by cAMP response element-binding protein (CREB) is required for cAMP-mediated inhibition of mitogen-activated protein kinase (MAPK) p38 activation in fibroblasts, which leads to suppression of NF-kappaB activity and promotion of tumor necrosis factor-alpha (TNF-alpha)-induced cell death. However, it remains unknown whether this regulation is also applicable to fibroblastoma cells. METHODS: Intracellular cAMP was determined in L929 fibroblastoma cells after treatment of the cells with various cAMP elevation agents. Effects of cAMP in the presence or absence of the RNA synthesis inhibitor actinomycin D or small interfering RNAs (siRNAs) against CREB on TNF-alpha-induced cell death in L929 cells were measured by propidium iodide (PI) staining and subsequent flow cytomety. The activation of p38 and c-Jun N-terminal protein kinase (JNK), another member of MAPK superfamily, was analyzed by immunoblotting. JNK selective inhibitor D-JNKi1 and p38 selective inhibitor SB203580 were included to examine the roles of JNK and p38 in this process. The expression of DLC or other mediators of cAMP was analyzed by immunoblotting. After ectopic expression of DLC with a transfection marker GFP, effects of cAMP on TNF-alpha-induced cell death in GFP+ cells were measured by PI staining and subsequent flow cytomety. RESULTS: Elevation of cAMP suppressed TNF-alpha-induced necrotic cell death in L929 fibroblastoma cells via CREB-mediated transcription. The pro-survival role of cAMP was associated with selective unresponsiveness of L929 cells to the inhibition of p38 activation by cAMP, even though cAMP significantly inhibited the activation of JNK under the same conditions. Further exploration revealed that the induction of DLC, the major mediator of p38 inhibition by cAMP, was impaired in L929 cells. Enforced inhibition of p38 activation by using p38 specific inhibitor or ectopic expression of DLC reversed the protection of L929 cells by cAMP from TNF-alpha-induced cell death. CONCLUSION: These data suggest that the lack of a pro-apoptotic pathway in tumor cells leads to a net survival effect of cAMP.