[Establishment of an Animal Model of Vesicoureteral Reflux Renal Injury through Partial Bladder Outlet Obstruction].

OBJECTIVE: To establish an animal model of reflux renal damage through bladder outlet obstruction. METHODS: Sixty male C57BL/6 mice aged 6-8 weeks were randomly assigned to a control group, a sham operation group, and a partial bladder outlet obstruction (PBOO) group, with 20 mice in each group. Laparotomy were performed on the PBOO mice under anesthesia in order to separate the bladder necks and to perform guided partial ligation of the bladder neck with a metal rod of 0.3 mm diameter. Mice in the sham operation group had laparotomy and had their bladder necks separated without ligation. The control group did not receive any treatment. 7 days after the surgery, 12 surviving mice were randomly selected from each group to observe the general changes of the bladder, ureter, renal pelvis and kidney. Retrograde urography was performed through the bladder. Kidney tissues were extracted for histopathological analysis. The expression levels of Vimentin, proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA) were examined with Western blot, immunohistochemistry and immunofluorescence staining tests, respectively. RESULTS: Compared with the control and sham operation group, the bladder, ureter, and renal pelvis of the mice in the PBOO group were significantly enlarged, vesicoureteral reflux was more obvious, the kidney volume and mass increased ( P<0.001), and renal parenchyma became thinner ( P<0.000 1). Histopathological staining showed glomerular atrophy, renal tubule expansion, tubulointerstitial inflammatory cell infiltration, glomerular basement membrane hyperplasia and obvious interstitial fibrosis. Western blot, immunofluorescence and immunohistochemistry staining showed that the expression levels of Vimentin, PCNA and α-SMA in kidney tissue were elevated ( P<0.000 1). CONCLUSION: After PBOO, the bladder, ureter, and kidney of the mice showed obvious morphological alteration and presented reflux renal fibrosis-like damage. This can be used as an animal model to study the pathological alteration mechanism and therapeutic measures of renal fibrosis caused by bladder outlet obstruction.

Migrant adolescents' behavioral problems compared to host adolescents and adolescents in their region of origin: a longitudinal study.

BACKGROUND: Since the 1990s, families from the ecologically hostile mountainous southern areas of Ningxia Province, China, have been migrating to the northern areas of the province. This study compared the prevalence of behavioral problems among migrant adolescents to those among host adolescents (adolescents from the northern areas) and adolescents in the region of origin (adolescents from the southern areas), to determine whether ecological migration is related to adolescent behavioral problems, and possible changes in such problems over time. METHODS: We used the Children and Adolescents Ecological Migration Survey on Mental Health, administered to 4805 students aged 12-16 years and their parents between 2012 and 2014 (W1), of whom 1753 students and their parents completed the follow-up between 2014 and 2017 (W2). Parents answered questions related to adolescent behavioral problems, main source of family income, parents' desire to reverse migrate, improved standard of living, and parents' educational attainment, while children completed the Eysenck Personality Questionnaire and a classroom environment questionnaire. RESULTS: The prevalence of behavioral problems among the migrant adolescents (28.04%) was significantly higher than among host adolescents (21.59%) or adolescents in the region of origin (24.37%; p < 0.001) at W1. After adjusting for gender and age, parents' work outside the home was the main source of family income (OR = 1.42, 95% CI = 1.13-1.78), and adolescents' learning burden (OR = 1.04, 95% CI = 1.01-1.06) in school negatively influenced behavioral problems. Strong student-teacher relationships (OR = 0.97,95% CI = 0.94-0.99) and parents who had no intention to move back to the original residence (OR = 0.70, 95% CI = 0.52-0.94) exerted a protective effect at W1; at W2, a protective effect was associated with improved living conditions (OR = 0.39-0.55, 95% CI = 0.25-0.84). The extent of behavioral problems among migrant adolescents significantly decreased after two years. CONCLUSION: Ecological migration will increase children's behavioral problems in the early stage, with various factors influencing the extent of these problems.

MiR-29a-3p Enhances the Viability of Rat Neuronal Cells that Injured by Oxygen-Glucose Deprivation/Reoxygenation Treatment Through Targeting TNFRSF1A and Regulating NF-κB Signaling Pathway.

OBJECTIVE: We attempt to investigate the role of TNFRSF1A and its underlying mechanism in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced injury in rat pheochromocytoma PC12 cells. METHODS: Public datasets GSE61616 and GSE106680 were downloaded from GEO database. PC12 cells were used to construct OGD/R models. QRT-PCR and western blot were implemented to test the relative mRNA and protein levels, respectively. The miRNA online prediction website TargetScan was used to predict TNFRSF1A upstream regulated miRNAs, which were then confirmed by luciferase reporter assay. The changes in cell viability and apoptosis were evaluated using cell counting kit 8 (CCK-8), lactose dehydrogenase (LDH), and flow cytometry assays. RESULTS: Bioinformatics analysis demonstrated that the expression of TNFRSF1A was upregulated in CI/RI and middle cerebral artery occlusion models compared with control, respectively. And a significant upregulation was also observed in OGD/R-damaged PC12 cells. Depletion of TNFRSF1A can notably enhance the cells proliferation after OGD/R treatment, while enlargement of TNFRSF1A presented the opposite outcomes. Moreover, miR-29a-3p was shown to be the upstream regulatory miRNA of TNFRSF1A. The levels of TNFRSF1A were inversely mediated by miR-29a-3p. Overexpression of miR-29a-3p can raise the cell viability, decrease the LDH activity, and reduce the apoptotic ratio in OGD/R-treated cells. Besides, TNFRSF1A can attenuate the protective effect of miR-29a-3p on OGD/R-treated cells. Furthermore, miR-29a-3p mimic inhibited, while overexpression of TNFRSF1A promoted the activation of NF-κB signaling pathway, and TNFRSF1A can attenuate the suppressive effect of miR-29a-3p on the NF-κB pathway. CONCLUSION: Our research illustrated that the potential regulatory role of miR-29a-3p/TNFRSF1A axis in neurons cells suffered from OGD/R, and their effects on NF-κB signaling pathway, providing a possible bio-target for protecting cells from OGD/R damage .

Association between Serum Fructosamine and Kidney Function in Nondiabetic Individuals without Chronic Kidney Disease.

BACKGROUND: Serum fructosamine (SF) has been considered to be an indicator that estimates glycemic control in patients with diabetes mellitus (DM). There is increasing evidence that SF concentration and oxidative stress are significantly elevated in patients with chronic kidney disease (CKD). However, the data about SF and its association with kidney function are lacking in nondiabetic individuals without CKD. We included 1891 nondiabetic individuals who had not been diagnosed with CKD to determine the association between SF and kidney function. MATERIAL AND METHODS: We conducted a retrospective analysis on the basis of the biochemistry database in nondiabetic individuals without CKD. RESULTS: When eligible participants were stratified in accordance with SF quartiles, from the bottom to the top quartile of SF, a significant decrease of estimated glomerular filtration rate (GFR) was observed in baseline data. SF concentration was negatively associated with estimated GFR (r=-0.066, P=0.004) in the Pearson correlation analysis. Estimated GFR was associated with SF levels independently of glucose (GLU), total cholesterol (TC), triglyceride (TG), and total protein (TP) in multivariable logistic regression analysis (OR=0.984; CI 95% 0.977-0.991; P<0.001). CONCLUSIONS: We suggest that mild elevation of SF concentration is associated with estimated GFR in nondiabetic individuals without CKD. These findings indicate that SF may underlie CKD in nondiabetic individuals.

[Clinical features and risk factors of co-morbid tic disorder in children with attention deficit hyperactivity disorder].

OBJECTIVE: To study the clinical features and risk factors of co-morbid tic disorder (TD) in children with attention deficit hyperactivity disorder (ADHD). METHODS: A total of 312 children with ADHD were involved in this study. Subtypes of co-morbid TD, incidences of TD in different subtypes of ADHD (ADHD-I, ADHD-HI and ADHD-C) were observed. Thirteen potential factors influencing the comorbidity rate of TD in ADHD were evaluated by univariate analysis and multiple logistic regression analysis. RESULTS: Forty-two of 312 children with ADHD suffered from co-morbid TD (13.5%). Comorbidity rate of TD in children with ADHD-C (24.1%) was significantly higher than in those with ADHD-HI (10.9%) and ADHD-I (8.8%) (P<0.05). There were 21 cases (50.0%) of transient TD, 12 cases (28.6%) of chronic TD, and 9 cases (21.4%) of Tourette syndrome. The univariate analysis revealed 6 factors associated with comorbidity: addiction to mobile phone or computer games, poor eating habits, infection, improper family education, poor relationship between parents and poor relationship with schoolmates. Multiple logistic analysis revealed two independent risk factors for comorbidity: improper family education (OR=7.000, P<0.05) and infection (OR=2.564, P<0.05). CONCLUSIONS: The incidence of co-morbid TD in children with ADHD is influenced by many factors, and early interventions should be performed based on the main risk factors.

Diabetic retinopathy identification based on multi-source-free domain adaptation.

AIM: To address the challenges of data labeling difficulties, data privacy, and necessary large amount of labeled data for deep learning methods in diabetic retinopathy (DR) identification, the aim of this study is to develop a source-free domain adaptation (SFDA) method for efficient and effective DR identification from unlabeled data. METHODS: A multi-SFDA method was proposed for DR identification. This method integrates multiple source models, which are trained from the same source domain, to generate synthetic pseudo labels for the unlabeled target domain. Besides, a softmax-consistence minimization term is utilized to minimize the intra-class distances between the source and target domains and maximize the inter-class distances. Validation is performed using three color fundus photograph datasets (APTOS2019, DDR, and EyePACS). RESULTS: The proposed model was evaluated and provided promising results with respectively 0.8917 and 0.9795 F1-scores on referable and normal/abnormal DR identification tasks. It demonstrated effective DR identification through minimizing intra-class distances and maximizing inter-class distances between source and target domains. CONCLUSION: The multi-SFDA method provides an effective approach to overcome the challenges in DR identification. The method not only addresses difficulties in data labeling and privacy issues, but also reduces the need for large amounts of labeled data required by deep learning methods, making it a practical tool for early detection and preservation of vision in diabetic patients.

[Analysis of the pregnancy outcomes of 13 723 tubes of sperm specimens from the sperm bank].

OBJECTIVE: To investigate the safety of assisted reproductive technology (ART) with donated sperm from the sperm bank and the differences in the pregnancy outcomes of different means of promoting pregnancy. METHODS: We analyzed and compared the feedback data on promoting pregnancy with donated sperm from the sperm bank by artificial insemination by donor (AID), in vitro fertilization (IVF), and intracytoplasm sperm injection (ICSI). RESULTS: Totally, 13 723 tubes of sperm specimens were used for ART. The number of specimens used differed in different clinical reproductive centers, some using 1 tube and others using 2 tubes per cycle. The 13 723 tubes were used for a total of 7 743 cycles. Among the 7 123 cycles of AID, there were 1 415 clinical pregnancies (19.87%), 1 221 normal births (86.29%), 169 abortions (11.94%), 6 cases of birth defects (0.43%), 19 ectopic pregnancies (1.34%), and 0 sexually transmitted infection. Among the 571 cycles of IVF, there were 367 clinical pregnancies (64.27%), 330 normal births (89.92%), 35 abortions (9.54%), 0 birth defect, 2 ectopic pregnancies (0.54%), and 0 sexually transmitted infection. Among the 49 cycles of ICSI, there were 28 clinical pregnancies (57.14%), 25 normal births (89.29%), 3 abortions (10.71%), 0 birth defect, 0 ectopic pregnancy, and 0 sexually transmitted infection. There were statistically significant differences in the rate of clinical pregnancy among AID, IVF and ICSI (P < 0.05), but not between IVF and ICSI (P > 0.05), nor were there any significant differences in the rates of abortion, birth defects and ectopic pregnancy among AID, IVF and ICSI (P > 0.05). CONCLUSION: None of the recipients of the donated sperm from the sperm bank was infected with sexually transmitted diseases. AID, IVF and ICSI showed no significant differences from natural conception in the rates of abortion, birth defects and ectopic pregnancy. ART with donated sperm from the sperm bank is safe. IVF and ICSI are associated with a higher rate of pregnancy than AID, though the latter costs less than the former two.

[Effect of methylation inhibitor on EphB4 gene expression, proliferation and apoptosis in CEM cells].

OBJECTIVE: To study the regulation of methylation inhibitor 5-aza-2'-deoxycytidine on transcription of EphB4 gene and effects on the proliferation and apoptosis of human acute lymphocyte leukemia cell line CEM. METHODS: Bisulfite sequencing PCR was used to detect CpG island methylation density in EphB4 promoter. The expression of EphB4 mRNA and protein was determined by Q-PCR and Western blot. MTS assay and flow cytometry were used to detect the apoptosis of CEM cells after treatment with different concentrations of 5-aza-2'-deoxycytidine (1.0, 2.5 and 5 µmol/L). RESULTS: Methylation of EphB4 gene promoter was detected in CEM cells (31.4%). The methylation level of EphB4 gene was down-regulated after treatment with various concentrations of 5-aza-2'-deoxycytidine. The EphB4 mRNA and protein expression in CEM cells increased after 5-aza-2'-deoxycytidine treatment. 5-Aza-2'-deoxycytidine significantly inhibited the cell growth in dose and time dependent manners. Early apoptosis rates of CEM cells increased from 4.1% to 24.8% 96 hrs after 5-aza-2'-deoxycytidine treatment. CEM cells in G1 phase decreased from 62.4% to 46.8%, cells in G2 phase increased from 2.1% to 16.2%, and CEM cells were arrested in G2 phase after treatment with 5 µmol/L 5-aza-2'-deoxycytidine for 96 hrs. CONCLUSIONS: 5-Aza-2'-deoxycytidine, an inhibitor of specific methylation transferase, can induce expression of the silent EphB4 gene in CEM cells, inhibit the proliferation of leukemia cells and induce cell apoptosis.

[Genetic polymorphism of GST gene in children with infectious mononucleosis and acute lymphocytic leukemia].

OBJECTIVE: To study the relationship between glutathione S-transferase genes GSTT1 and GSTM1 polymorphisms and the susceptibility to infectious mononucleosis (IM) and acute lymphocytic leukemia (ALL) in children. METHODS: The case-control study involved 106 children with IM, 41 children with ALL and a control group of 100 children with non-hematologic and nontumorous diseases. The genetic polymorphisms of GSTT1 and GSTM1 were detected with multiplex polymerase chain reaction (PCR). Distribution of the genotypes in the children was analyzed. RESULTS: The frequency of GSTT1 null genotype in children with IM was significantly higher than in the control group (P<0.05). The risk of IM in children carrying GSTT1 null genotype was 2.186 times higher than in those carrying GSTT1 non-null genotype. The children carrying both GSTT1 and GSTM1 null genotype had a higher risk of suffering from IM compared to those carrying only one of the null genotypes (OR=4.937). The frequency of GSTM1 null genotype in children with ALL was significantly higher than in the control group (P<0.05). The risk of ALL in children carrying GSTM1 null genotype was 2.242 times higher than in those in carrying GSTT1 non-null genotype. Children carrying both GSTT1 and GSTM1 null genotype had a higher risk of suffering from ALL compared with those carrying only one of the null genotypes (OR=8.552). CONCLUSIONS: Children carrying GSTT1 or GSTM1 null genotype have a high risk of suffering from IM or ALL. Still more increased susceptibility to IM or ALL may occur in children who carry both GSTT1 and GSTM1 null genotype. GSTT1 and GSTM1 might play a potential role in the pathogenesis of both IM and ALL.

The novel HLA-B*35:501 allele, identified by sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-B*35:501 differs from HLA-B*35:02:01:01 by three nucleotides in exon 2.

[Effects of eye acupuncture on motor evoked potential and somatosensory evoked potential in patients with incomplete spinal cord injury based on neuroelectrophysiological technology].

OBJECTIVE: To observe the effects of eye acupuncture on motor evoked potential (MEP) and somatosensory evoked potential (SEP) in the patients with incomplete spinal cord injury so as to evaluate its clinical efficacy. METHODS: According to the random number table, 90 patients were divided into exercise therapy group, eye acupuncture group and eye acupuncture combined exercise therapy group (combined treatment group), 30 cases in each. In the exercise therapy group, patients were treated with the routine exercise and occupational therapy. Patients of the eye acupuncture group were treated with eye acupuncture at upper jiao region, lower jiao region, liver region and kidney region bilaterally. Patients of the combined treatment group were given the routine exercise and occupational therapy combined with eye acupuncture. All the treatments were conducted once daily, 7 days as one treatment course for 4 treatment courses. Before treatment and 4 weeks after treatment, the motor function, light touch sensation and pinprick sensation, injury grade and clinical efficacy were assessed separately, using the criteria developed by the American Spinal Injury Association. The modified Barthel index(MBI) was adopted to evaluate the activities of daily livings. By monitoring SEP and MEP, the neurophysiological conditions were assessed for spinal cord injury. RESULTS: The total effective rate was 56.7% (17/30), 66.7% (20/30) and 90.0% (27/30) in the exercise therapy group, the eye acupuncture group and the combined treatment group, respectively. The total effective rate in the combined treatment group was higher than those in the other two groups (P<0.05). Compared with those before treatment, the scores of motor function, light tough sensation and pinprick sensation were all increased after treatment in three groups (P<0.05), MBI score was increased in both the exercise therapy group and the combined treatment group (P<0.05), and the latency of SEP (N11, N20, N23, P38) and the Cortical (hand region), Csp, Cortical (leg region) and Lsp of MEP were all shortened in the three groups separately (P<0.05). After treatment, compared with the exercise therapy group, the score of motor function was increased (P<0.05), MBI score decreased (P<0.05) and MEP latency shortened (P<0.05) in the eye acupuncture group. After treatment, compared with the exercise therapy group and the eye acupuncture group, the scores of motor function, light touch sensation and pinprick sensation, as well as MBI score were all increased (P<0.05), and the latency of SEP (N11,N20,N23,P38) and MEP shortened (P<0.05) in the combined treatment group. CONCLUSION: In treatment of incomplete spinal cord injury, eye acupuncture combined with exercise therapy can significantly increase the excitability of sensory and motor nerve conduction in the spinal cord and cerebral cortex of patients, effectively promote the recovery of patients' motor and sensory function and improve the activities of daily living.

The Regulatory Network and Role of the circRNA-miRNA-mRNA ceRNA Network in the Progression and the Immune Response of Wilms Tumor Based on RNA-Seq.

Circular RNA (circRNA), which is a newly discovered non-coding RNA, has been documented to play important roles in miRNA sponges, and the dysregulation of which is involved in cancer development. However, circRNA expression profiles and their role in initiation and progression of Wilms tumor (WT) remain largely unclear at present. Here, we used paired WT samples and high-throughput RNA sequencing to identify differentially expressed circRNAs (DE-circRs) and mRNAs (DE-mRs). A total of 314 DE-circRs and 1612 DE-mRs were identified. The expression of a subset of differentially expressed genes was validated by qRT-PCR. A complete circRNA-miRNA-mRNA network was then constructed based on the common miRNA targets of DE-circRs and DE-mRs identified by miRanda prediction tool. The Gene set enrichment analysis (GSEA) indicated that several signaling pathways involving targeted DE-mRs within the ceRNA network were associated with cell cycle and immune response, which implies their participation in WT development to some extent. Subsequently, these targeted DE-mRs were subjected to implement PPI analysis and to identify 10 hub genes. Four hub genes were closely related to the survival of WT patients. We then filtered prognosis-related hub genes by Cox regression and least absolute shrinkage and selection operator (LASSO) regression analysis to construct a prognosis-related risk score system based on a three-gene signature, which showed good discrimination and predictive ability for WT patient survival. Additionally, we analyzed the mutational landscape of these genes and the associations between their expression levels and those of immune checkpoint molecules and further demonstrated their potential impact on the efficacy of immunotherapy. qRT-PCR and western blotting (WB) analysis were used to validate key differentially expressed molecules at the RNA and protein levels, respectively. Besides these, we selected a key circRNA, circEYA1, for function validation. Overall, the current study presents the full-scale expression profiles of circRNAs and the circRNA-related ceRNA network in WT for the first time, deepening our understanding of the roles and downstream regulatory mechanisms of circRNAs in WT development and progression. We further constructed a useful immune-related prognostic signature, which could improve clinical outcome prediction and guide individualized treatment.

Programmed intermittent epidural bolus in parturients: A meta-analysis of randomized controlled trials.

BACKGROUND: To evaluate the efficacy and safety of programmed intermittent epidural bolus (PIEB) in parturients. METHODS: The PubMed, Embase, and the Cochrane Library (from inception to July 2021) were searched for identification of randomized placebo-controlled trials in which PIEB was applied in parturients. The outcomes were the effect of analgesia, satisfaction score, mode of delivery, duration of labor, neonatal condition, and adverse events. The pooled odds ratios (OR), weighted mean difference (WMD), and 95% confidence intervals (CIs) were calculated using random- and fixed-effects models. RESULTS: PIEB was found to be associated with decreased total consumption of ropivacaine (WMD = -15.83, 95% CI: -19.06 to -12.60, P < .00001; I2 = 61%; P for heterogeneity = .04), total consumption of sufentanil (WMD = -4.93, 95% CI: -6.87 to 2.98, P < .00001; I2 = 68%; P for heterogeneity = .05), numbers of patients who require patient-controlled epidural analgesia bolus (OR = 0.27, 95% CI: 0.14-0.51, P < .0001; I2 = 65%; P for heterogeneity = .01), the number of attempts (WMD = -4.12, 95% CI: -7.21 to -1.04, P = .009; I2 = 100%; P for heterogeneity < .00001), rate of breakthrough pain (OR = 0.47, 95% CI: 0.28-0.80, P = .005; I2 = 47%; P for heterogeneity = .09). Eight studies focus on the duration of analgesia. After by meta-analysis, we found that the pain visual analogue scale (VAS) score at 30 minutes, 2 hours, 4 hours, and 5 hours in PIEB group was significantly lower when compared with control group, (WMD = -0.15, 95% CI: -0.26 to -0.04, P = .006; I2 = 0%; P for heterogeneity = .64), (WMD = -0.79, 95% CI: -1.32 to 0.25, P = .004; I2 = 97%; P for heterogeneity < .00001), (WMD = -1.00, 95% CI: -1.08 to -0.91, P < .00001; I2 = 0%; P for heterogeneity = .67), (WMD = -1.81, 95% CI: -3.23 to -0.39, P = .01; I2 = 98%; P for heterogeneity < .00001), respectively. Nineteen studies discussed the mode of delivery between 2 groups. The results suggest that the rate of normal delivery is significantly higher in PIEB group compared with control group (OR = 1.37, 95% CI: 1.08-1.75, P = .01). The time of first and second stage of labor are significantly shorter in PIEB group compared with control group, the result is (WMD = -10.52, 95% CI: -14.74 to 4.76, P < .00001; I2 = 0%; P for heterogeneity = .86), (WMD = -1.48, 95% CI: -2.26 to -0.69, P = .0002; I2 = 35%; P for heterogeneity = .10), respectively. Thirteen studies concerned the satisfaction score of patients. The satisfaction score of patients in the PIEB group was significantly higher when compared with control group (WMD = 0.91, 95% CI: 0.42-1.39, P = .0003; I2 = 98%; P for heterogeneity < .00001). The Apgar score at 1, 5 minutes in PIEB group are significantly higher (WMD = 0.07, 95% CI: 0.02-0.13 P = .007; I2 = 55%; P for heterogeneity = .04), (WMD = -0.08, 95% CI: -0.12 to -0.05, P < .00001; I2 = 21%; P for heterogeneity = .27), respectively. CONCLUSIONS: PIEB is a good alternative for labor analgesia with better analgesic effect, maternal and infant outcome.

A novel cuproptosis-related subtypes and gene signature associates with immunophenotype and predicts prognosis accurately in neuroblastoma.

Background: Neuroblastoma (NB) is the most frequent solid tumor in pediatrics, which accounts for roughly 15% of cancer-related mortality in children. NB exhibited genetic, morphologic, and clinical heterogeneity, which limited the efficacy of available therapeutic approaches. Recently, a new term 'cuproptosis' has been used to denote a unique biological process triggered by the action of copper. In this instance, selectively inducing copper death is likely to successfully overcome the limitations of conventional anticancer drugs. However, there is still a gap regarding the role of cuproptosis in cancer, especially in pediatric neuroblastoma. Methods: We characterized the specific expression of cuproptosis-related genes (CRGs) in NB samples based on publicly available mRNA expression profile data. Consensus clustering and Lasso-Cox regression analysis were applied for CRGs in three independent cohorts. ESTIMATE and Xcell algorithm was utilized to visualize TME score and immune cell subpopulations' relative abundances. Tumor Immune Dysfunction and Exclusion (TIDE) score was used to predict tumor response to immune checkpoint inhibitors. To decipher the underlying mechanism, GSVA was applied to explore enriched pathways associated with cuproptosis signature and Connectivity map (CMap) analysis for drug exploration. Finally, qPCR verified the expression levels of risk-genes in NB cell lines. In addition, PDHA1 was screened and further validated by immunofluorescence in human clinical samples and loss-of-function assays. Results: We initially classified NB patients according to CRGs and identified two cuproptosis-related subtypes that were associated with prognosis and immunophenotype. After this, a cuproptosis-related prognostic model was constructed and validated by LASSO regression in three independent cohorts. This model can accurately predict prognosis, immune infiltration, and immunotherapy responses. These genes also showed differential expression in various characteristic groups of all three datasets and NB cell lines. Loss-of-function experiments indicated that PDHA1 silencing significantly suppressed the proliferation, migration, and invasion, in turn, promoted cell cycle arrest at the S phase and apoptosis of NB cells. Conclusions: Taken together, this study may shed light on new research areas for NB patients from the cuproptosis perspective.

Immune-related gene signature associates with immune landscape and predicts prognosis accurately in patients with Wilms tumour.

Wilms tumour (WT) is the most common kidney malignancy in children. Chemoresistance is the leading cause of tumour recurrence and poses a substantial therapeutic challenge. Increasing evidence has underscored the role of the tumour immune microenvironment (TIM) in cancers and the potential for immunotherapy to improve prognosis. There remain no reliable molecular markers for reflecting the immune landscape and predicting patient survival in WT. Here, we examine differences in gene expression by high-throughput RNA sequencing, focused on differentially expressed immune-related genes (IRGs) based on the ImmPort database. Via univariate Cox regression analysis and Lasso-penalized Cox regression analysis, IRGs were screened out to establish an immune signature. Kaplan-Meier curves, time-related ROC analysis, univariate and multivariate Cox regression studies, and nomograms were used to evaluate the accuracy and prognostic significance of this signature. Furthermore, we found that the immune signature could reflect the immune status and the immune cell infiltration character played in the tumour microenvironment (TME) and showed significant association with immune checkpoint molecules, suggesting that the poor outcome may be partially explained by its immunosuppressive TME. Remarkably, TIDE, a computational method to model tumour immune evasion mechanisms, showed that this signature holds great potential for predicting immunotherapy responses in the TARGET-wt cohort. To decipher the underlying mechanism, GSEA was applied to explore enriched pathways and biological processes associated with immunophenotyping and Connectivity map (CMap) along with DeSigN analysis for drug exploration. Finally, four candidate immune genes were selected, and their expression levels in WT cell lines were monitored via qRT-PCR. Meanwhile, we validated the function of a critical gene, NRP2. Taken together, we established a novel immune signature that may serve as an effective prognostic signature and predictive biomarker for immunotherapy response in WT patients. This study may give light on therapeutic strategies for WT patients from an immunological viewpoint.

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.

Photonic ultrawideband signal generator using an optically injected chaotic semiconductor laser.

We propose and demonstrate a method to generate ultrawideband (UWB) signals in the optical domain based on the chaotic dynamics of an optically injected semiconductor laser with optical feedback. The chaotic-UWB pulses with a fractional bandwidth of 116% and central frequency of 6.88 GHz are experimentally generated by controlling the injection strength and frequency detuning of the chaotic laser. The spectrum of the UWB signals is in full compliance with the Federal Communications Commission spectral mask, and the experimental results are qualitatively consistent with the simulated results.

A therapeutic strategy for choroidal neovascularization based on recruitment of mesenchymal stem cells to the sites of lesions.

Choroidal neovascularization (CNV) is a common cause of severe and irreversible visual loss; however, the treatment of CNV has been hindered by its complex and poorly understood pathogenesis. It has been postulated that bone marrow (BM)-derived cells (BMCs) contribute to CNV, but little is known about the role of mesenchymal stem cells (MSCs) in CNV and their therapeutic potential for CNV treatment. We found that BM-derived MSCs transplanted by intravenous injection into laser-induced CNV mouse models were specifically recruited into CNV lesions, where they differentiated into multiple cell types and participated in the development of neovascularization, without stagnation in other organs. By taking advantage of this recruitment potential, engineered MSCs were used to produce the antiangiogenic pigment epithelial-derived factor (PEDF) at the CNV sites, thereby inhibiting the growth of CNVs and stimulating regressive features. Further studies indicated that the effect may be mediated, at least partly, by retinal pigment epithelial (RPE) cells, which function as important regulators for CNV development. These results suggest that MSCs contribute to CNV and could serve as delivery vehicles of antiangiogenic agents for the treatment of a range of CNV-associated diseases.

Monocyte/macrophages promote vasculogenesis in choroidal neovascularization in mice by stimulating SDF-1 expression in RPE cells.

BACKGROUND: Monocyte-macrophages play important roles in choroidal neovascularization (CNV); however, the mechanism is unclear. This study investigated the effects of monocyte depletion on laser-induced CNV in mice, especially the involvement of bone marrow-derived cells (BMCs) and underlying molecular mechanisms. METHODS: Clodronate-liposomes (lip) were used to deplete monocytes and their effect on retinal pigmental epithelium (RPE) cells, endothelial cells, and BMCs was analyzed. Green fluorescent protein (GFP)-chimeric mice were developed by transplanting bone marrow cells from GFP transgenic mice to C57BL/6 J mice. CNV was induced by laser photocoagulation. Chimeric mice were intravenously treated with clodronate-lip, PBS-lip or PBS, 1 day before and after lasering. Histopathological and choroidal flatmount analysis were performed to measure CNV severity and BMCs recruitment. BMCs expression of endothelial cell marker CD31 and vascular smooth muscle cell marker α-SMA in CNV were detected by immunofluorescence. Expression of stromal cell-derived factor-1 (SDF-1) protein in vivo was detected by immunofluorescence as well as ELISA assay. SDF-1 was also examined by RT-PCR and ELISA in a human monocytes-RPE cells co-culturing system. RESULTS: No valid evidence for the toxicity of clodronate-lip was found. Depletion led to significant inhibition of CNV and BMCs recruitment into laser spots on days 3 and 14, reduced BMC expression of CD31 and α-SMA on day 14, and decreased expression of SDF-1 in vivo on day 3. SDF-1 was mostly within and around the RPE cells in the laser lesion. SDF-1 was dramatically up-regulated in RPE cells after co-culturing with monocytes. CONCLUSIONS: Monocytes may promote experimental CNV, especially BMC contribution in mice, by promoting SDF-1 production in RPE cells.

[Effects of cerebellar fastigial nucleus electrical stimulation on telomerase reverse transcriptase expression and mitochondrial apoptotic pathway in rats with focal cerebral ischemia and reperfusion].

OBJECTIVE: To study the effects of cerebellar fastigial nucleus (FN) electrical stimulation on telomerase reverse transcriptase expression and mitochondrial apoptotic pathway in rats with focal cerebral ischemia and reperfusion. METHODS: A total of 100 adult male Wistar rats were randomly divided into 3 groups: sham operation group, modeling group (2-hour cerebral ischemia, followed by 24, 48 & 72-hour reperfusion) and FN-stimulating group (electrical stimulation of FN for 1-hour one day before 2-hour cerebral ischemia, followed by 24, 48 & 72-hour reperfusion). HE (hematoxylin and eosin) and TTC (triphenyl tetrazolium chloride) staining were used to observe the morphological changes in rat brain and measure the ischemic lesion volumes. The expressions of TERT (telomerase reverse transcriptase) and Bax were detected by immunohistochemical methods and apoptotic cells by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling). The co-expression of TERT and Bax was detected by immunofluorescence double-labeling plus laser confocal microscopy. RESULTS: The morphological changes in rat brain were less greater in the FN-stimulating group than those in the modeling group. And the size of the cerebral infarct was significantly smaller in the FN-stimulating group (78.1 ± 2.9, 83.1 ± 4.5, 83.7 ± 4.8) than that in the modeling group (120.9 ± 8.2, 137.0 ± 4.2, 141.1 ± 3.3) (P < 0.05) at all reperfusion time points. As compared with the modeling group (16.1 ± 2.7, 16.9 ± 2.4, 11.6 ± 3.5), the FN-stimulating group (31.1 ± 3.5, 30.0 ± 3.4, 18.9 ± 3.3) had a significantly larger number of TERT-positive cells (P < 0.05) and a significantly reduced number of TUNEL-positive cells (49.6 ± 2.8, 67.0 ± 3.7, 46.8 ± 3.2 vs 40.2 ± 3.1, 54.8 ± 2.8, 37.3 ± 2.4) (P < 0.05). The number of Bax-positive cells at different reperfusion time points in the FN-stimulating group was not significantly different from those in the modeling group (P > 0.05). TERT partially co-localized with Bax in the cytoplasm. The number of double-labeled cells was significantly higher in the FN-stimulating group than that in the modeling group (14.1 ± 1.3, 12.9 ± 2.4, 9.0 ± 2.0 vs 8.2 ± 1.1, 6.3 ± 2.4, 6.0 ± 2.9) (P < 0.05). CONCLUSION: The expression of TERT significantly increases after a stimulation of FN. TERT may bind to Bax and inhibit Bax-mediated apoptosis by suppressing the mitochondrial relocalization of Bax from cytosol.