Serum troponin I: a potential biomarker of hypoxic-ischemic encephalopathy in term newborns.

OBJECTIVE: This study was intended to evaluate the predictive values of serum procalcitonin (PCT), lactate, creatine kinase (CK-MB), and troponin I on the diagnosis and staging of neonatal hypoxic-ischemic encephalopathy (HIE). MATERIALS AND METHODS: We retrospectively retrieved data from electronic medical records at our children's hospital, and we included all term newborns admitted between December 2018 and June 2020 with features of perinatal asphyxia. Receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to measure and evaluate the predictive values of biomarkers. p values < 0.05 were set as statistical significance. RESULTS: A total of 201 neonates were included. They were grouped as control (n = 40), mild HIE (n = 105), moderate HIE (n = 36), and severe HIE (n = 20). Serum lactate, PCT, CK-MB, and troponin I levels in severe hypoxic-ischemic brain injury group were significantly higher than those in mild to moderate hypoxic-ischemic brain injury group and control group (p < 0.05). Based on ROC and AUC analysis, troponin I showed highest predictive ability with AUC of 0.904, and sensitivity and specificity of 95.00% and 87.50% respectively. CONCLUSION: Serum troponin I has a good predictive value for neonatal hypoxic-ischemic encephalopathy after perinatal asphyxia.

Identification of Novel Mutations in Chinese Infants With Citrullinemia.

Citrullinemia is a rare autosomal recessive disorder characterized by elevated concentrations of citrulline in the blood resulting from malfunction of the urea cycle. It is categorized into two types, types I and II, which are caused by argininosuccinate synthase 1 (ASS1), and citrin (SLC25A13) gene mutations, respectively. In this study, we performed genetic analysis on nine Chinese infants with citrullinemia using next-generation sequencing, which identified a novel mutation (p.Leu313Met) and a rare mutation (p.Thr323Ile, rs1250895424) of ASS1. We also found a novel splicing mutation of SLC25A13: c.1311 + 4_+7del. Functional analysis of the ASS1 missense mutations showed that both significantly impaired the enzyme activity of ASS1, with the p. Thr323Ile mutation clearly affecting the interaction between ASS1 and protein arginine methyltransferase 7 (PRMT7). These findings expand the mutational spectrum of ASS1 and SLC25A13, and further our understanding of the molecular genetic mechanism of citrullinemia in the Chinese population.

Risk factors and prediction score model for unplanned readmission among neonates with NRDS under one year of age: A retrospective cohort study.

Objective: This study aimed to analyze the risk factors and establish a prediction score model for unplanned readmission among neonates with neonatal respiratory distress syndrome (NRDS) for respiratory problems under one year of age. Methods: This retrospective cohort study enrolled 230 neonates with NRDS who were admitted between January 2020 and December 2020. The infants were classified into two subgroups based on whether they were readmitted for respiratory problems under one year of age: readmit group and non-readmit group. Readmission risk factors for NRDS were analyzed by logistic regression and a prediction score model was generated. Results: Among the 230 enrolled infants, 51 (22%) were readmitted, and 179 (78%) were not readmitted. In univariate analysis, compared with non-readmit group infants, readmit group infants had a significantly younger birth gestational age (31.9 ± 2.3 vs. 32.8 ± 2.5 weeks, p = 0.012), lower birth weight (1,713.7 ± 501.3 g vs. 1,946.8 ± 634.4 g, p = 0.007), older age at discharge (41.7 vs. 31.7 days, p = 0.012), higher proportion of necrotizing enterocolitis (NEC) (31% vs. 16%, p = 0.016), higher rate of blood transfusion (39% vs. 25%, p = 0.049), higher rate of postnatal dexamethasone (DEX) administration (28% vs. 9.5%, p = 0.001), and higher rate of home oxygen therapy (HOT) (57% vs. 34%, p = 0.003). Moreover, readmit group infants had significantly longer antibiotic days usage (12.0 vs. 10.0 days, p = 0.026) and a longer duration of hospital stay (41.0 vs. 31.0 days, p = 0.012) than non-readmit group infants. The multivariate logistic regression analysis showed that taking readmission as a target variable, postnatal DEX administration (OR: 2.689, 95% CI: 1.168-6.189, p = 0.020), HOT (OR: 2.071, 95% CI: 1.060-4.046, p = 0.033), and NEC (OR: 2.088, 95% CI: 0.995-4.380, p = 0.051) could be regarded as risk factors for readmission. A scoring model predicting readmission was administered with a positive predictive value of 0.651 (95% CI: 0.557-0.745, p = 0.002), with a sensitivity of 0.412 and a specificity of 0.888 at a cut-off of 3.5 points, which were evaluated on the receiver operating characteristic curve. Conclusions: Postnatal DEX administration, HOT, and NEC were risk factors for readmission of NRDS. NRDS infants with a predictive score of 3.5 points or more were at high risk for unplanned readmission.

Impact of the COVID-19 pandemic on neonatal admissions in a tertiary children's hospital in southwest China: An interrupted time-series study.

BACKGROUND: The unprecedented coronavirus disease 2019 (COVID-19) pandemic has caused millions of infections worldwide and represents a significant challenge facing modern health care systems. This study was conducted to investigate the impact of lockdown measures in a tertiary Children's Hospital in southwest China, which might be used to predict long-term effects related to health-seeking behavior of parents/caregivers. METHODS: This study included newborns enrolled over a span of 86 weeks between January 4, 2019, and August 27, 2020. We designated two time periods for analysis purposes: a stable pre-COVID period(55 weeks between January 4, 2019, and January 23, 2020) and a COVID-impacted period (31 weeks between January 24, 2020, and August 27, 2020). An interrupted time-series analysis was employed to compare changes and trends in hospital admissions and disease spectra before and after the period of nonpharmaceutical interventions (NPIs). Furthermore, this study was conducted to evaluate whether the health-seeking behavior of parents/caregivers was influenced by pandemic factors. RESULTS: Overall, 16,640 infants were admitted to the neonatology department during the pre-COVID period (n = 12,082) and the COVID-impacted period (n = 4,558). The per week neonatal admissions consistently decreased following the first days of NPIs (January 24, 2020). The average weekly admission rates of 220/week pre-COVID period and 147/week COVID-impacted period. There was an evident decrease in the volume of admissions for all disease spectra after the intervention, whereas the decrease of patients complaining about pathological jaundice-related conditions was statistically significant (p<0.05). In the COVID-impacted period, the percentage of patients who suffered from respiratory system diseases, neonatal encephalopathy, and infectious diseases decreased, while the percentage of pathological jaundice-related conditions and gastrointestinal system diseases increased. The neonatal mortality rates (NMRs) increased by 8.7% during the COVID-impacted period compared with the pre-COVID period. CONCLUSIONS: In summary, there was a significant decline in neonatal admissions in a tertiary care hospital during the COVID-19 Pandemic and the associated NPIs. Additionally, this situation had a remarkable impact on disease spectra and health-seeking behavior of parents/caregivers. We, therefore, advise continuing follow-ups and monitoring the main health indicators in vulnerable populations affected by this Pandemic over time.

Inflammatory stress exacerbates lipid-mediated renal injury in ApoE/CD36/SRA triple knockout mice.

Both lipids and inflammation play important roles in the progression of kidney disease. This study was designed to investigate whether inflammation exacerbates lipid accumulation via LDL receptors (LDLr), thereby causing renal injury in C57BL/6J mice, apolipoprotein E (ApoE) knockout (KO) mice, and ApoE/CD36/scavenger receptor A triple KO mice. The mice were given a subcutaneous casein injection to induce inflammatory stress. After 14 wk, terminal blood samples were taken for renal function, lipid profiles, amyloid A (SAA), and IL-6 assays. Lipid accumulation in kidneys was visualized by oil red O staining. Fibrogenic molecule expression in kidneys was examined. There was a significant increase in serum SAA and IL-6 in the all casein-injected mice compared with respective controls. Casein injection reduced serum total cholesterol, LDL cholesterol, and HDL cholesterol and caused lipid accumulation in kidneys from three types of mice. The expression of LDLr and its regulatory proteins sterol-responsive element-binding protein (SREBP) 2 and SREBP cleavage-activating protein (SCAP) were upregulated in inflamed mice compared with controls. Casein injection induced renal fibrosis accompanied by increased expression of fibrogenic molecules in the triple KO mice. These data imply that inflammation exacerbates lipid accumulation in the kidney by diverting lipid from the plasma to the kidney via the SCAP-SREBP2-LDLr pathway and causing renal injury. Low blood cholesterol levels, resulting from inflammation, may be associated with high risk for chronic renal fibrosis.

Comparative efficacy of anti-epileptic drugs for neonatal seizures: A network meta-analysis.

BACKGROUND: Anti-epileptic drugs have different effects on neonatal seizures, and new agents have been widely used in recent years. Meanwhile, significant differences still exist in the treatment for neonatal seizures, whether in choice of drug or in duration of treatment. And with the increase in options for treatment, the best choice of second-line treatment has not been recommended. METHODS: The MEDLINE, the Cochrane Library, Web of Science, Embase and clinicaltrials.gov databases were searched (January 1, 1960 to October 20, 2020). Randomized controlled trials (RCTs) or observational investigations studying anti-epileptic drugs for neonatal seizures were selected. And then we conducted a network meta-analysis and examined comparative efficacy of the first-line and second-line anti-epileptic drugs for neonatal seizures. RESULTS: Data were extracted from 11 included studies by 2 independent investigators. Random effects models were used to estimate odds ratios (ORs). We performed direct meta-analyses with a random effects model and network meta-analyses for first-line and second-line drugs. Five published RCTs and 6 observational investigations with 1333 patients and 6 interventions contributed to the analysis. CONCLUSION: We recommend phenobarbital as the first-line drug for neonatal seizures. In addition, there is a tendency for levetiracetam to be an effective second-line treatment for neonatal seizures after failure of first-line drugs.

[Inflammation enhances the accumulation of lipid in ApoE/SRA/CD36 KO mice liver].

OBJECTIVE: To investigate if inflammatory stress enhances liver lipid accumulation via SREBPs mediated dysregulation of low density protein receptor (LDLr) expression in apolipoprotein E, scavenger receptors class A and CD36 triple knockout (ApoE/SRA/CD36 KO) mice. METHODS: 16 Male ApoE/SRA/CD36 KO mice were subcutaneously injected with 0.5 ml 10% casein or PBS. The mice were fed a Western diet (Harlan, TD88137) containing 21% fat and 0.15% of cholesterol for 14 weeks. Animals were sacrificed and blood samples were collected. The serum amyloid A (SAA), IL-6, total cholesterol (TC), LDL and high density protein (HDL) were assayed. The lipid accumulation in liver was evaluated by Oil Red O staining. The mRNA and protein expression of SREBP-2, SREBPs cleavage activating protein (SCAP) and LDLr were analyzed by Real-Time Polymerase Chain Reaction (RT-PCR) and immunohistochemistry staining. RESULTS: Blood levels of SAA [(26.60+/-3.24) ng/ml vs (14.35+/-1.73) ng/ml, P < 0.01] and IL-6 [(36.37+/-2.20) pg/ml vs (18.02+/-4.87) pg/ml, P < 0.01] were higher, while TC [(7.72+/-1.70) mmol/L vs (13.23+/-3.61)mmol/L, P less than 0.01], LDL-cholesterol [(2.94+/-0.44) mmol/L vs (9.28+/-3.66) mmol/L, P less than 0.01] and HDL cholesterol [(2.24+/-0.63) mmol/L vs (4.13+/-0.42) mmol/L, P less than 0.01] were lower in inflamed mice compared to controls. ORO staining showed that lipid accumulation in the liver was more extensive in inflamed group despite lower blood lipid levels. Meanwhile, Real Time PCR data showed inflammation induced the expression of LDLr (4.56 fold), SCAP (3.14 fold) and SREBP-2 (14.72 fold) in liver. Immunohistochemical staining also indicated increased proteins expression in the liver, which was consistent with mRNA data. CONCLUSIONS: Inflammation causes lipid accumulation in liver via disrupting SREBP-2 and LDLr expression.

Molecular characterization and expression analysis of the autophagic gene beclin 1 from the purse red common carp (Cyprinus carpio) exposed to cadmium.

Beclin 1, the mammalian orthologue of yeast Atg6, has a central role in autophagy, which has been linked to diverse biological processes including immunity, development, tumor suppression, lifespan extension, etc. However, the relevant study about Beclin 1 is rare in fish compared with mammals. In this study, we isolated Beclin 1 gene from the kidney tissue of common carp (Cyprinus carpio) using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of cloned Beclin 1 comprised 447 amino acids, which showed approximately 80.7% identity and 88.9% similarity to human Beclin 1. It possessed a typical Bcl-2 homology domain 3 (BH3) and an evolutionarily conserved domain (ECD). Phylogenetic analysis demonstrated that common carp Beclin 1 formed a clade with zebrafish Beclin 1. To explore the relationship between Beclin 1 and cadmium (Cd)-induced injury, a Cd exposure experiment was conducted. The result showed that Cd content was significantly increased in a dose-dependent manner in kidney after Cd exposure. Swelling and vacuolation of renal tubular epithelial cells, and glomerular hyalinization were observed. Renal leukocyte infiltration was diffusely distributed in the interstitial tissue. Real-time quantitative RT-PCR analysis revealed that the mRNA transcript level of Beclin 1 was markedly up-regulated in a dose-dependent and time-dependent manner after exposure to Cd. Similarly, Western blot analysis indicated that its protein level was significantly elevated in a dose-dependent manner after Cd treatment. All the results indicate that the common carp Beclin 1 gene may play a regulatory role against Cd toxicity.

Molecular cloning, immunohistochemical localization, characterization and expression analysis of caspase-9 from the purse red common carp (Cyprinus carpio) exposed to cadmium.

Caspase-9, the essential initiator caspase is believed to play a central role in mitochondria-mediated apoptosis signaling. In this study, we isolated the caspase-9 gene from common carp, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of caspase-9, composed of 436 amino acids, showed approximately 47.6% identity and 64.7% similarity to human caspase-9. It also possessed a conserved caspase-associated recruitment domain (CARD), a large subunit and a small subunit. Phylogenetic analysis clearly demonstrated that caspase-9 formed a clade with cyprinid fish caspase-9. Real-time quantitative PCR analysis revealed that caspase-9 transcripts were not significantly increased in kidney after exposure to cadmium (Cd). Whereas caspase-9 cleaved fragments were detected using Western blot analysis with the same Cd treatment condition. Furthermore, the result of immunohistochemical detection showed immunoreactivities were predominantly limited to the cytoplasm of renal tubular epithelial cells and no remarkable changes of immunopositive staining were observed after Cd treatment. Accordingly, the results signify that caspase-9 may play an essential role in Cd induced apoptosis.

Cadmium triggers kidney cell apoptosis of purse red common carp (Cyprinus carpio) without caspase-8 activation.

Caspase-8, the essential initiator caspase, is believed to play a pivotal role in death receptor-mediated apoptotic pathway. It also participates in mitochondria-mediated apoptosis via cleavage of proapoptotic Bid in mammals. However, its role in fish remains elusive in Cadmium-induced apoptotic pathway. In this study, we isolated the caspase-8 gene from common carp, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of caspase-8 comprised 475 amino acids, which showed approximately 64.1% identity and 79.8% similarity to zebrafish (Danio rerio) caspase-8, possessed two conserved death effector domains, a large subunit and a small subunit. Phylogenetic analysis demonstrated that caspase-8 formed a clade with zebrafish caspase-8. In kidney, cadmium (Cd) exposure triggered apoptosis and increased caspase-3 and -9 activities, whereas it did not affect caspase-8 activity. Real-time quantitative PCR analysis revealed that caspase-8 transcriptional level was not significantly increased in kidney after exposure to Cd. Using Western blot analysis, no caspase-8 cleaved fragment was detected and no significant alteration of procaspase-8 level was found with the same Cd-treated condition. Moreover, the immunopositive staining was predominantly limited to the cytoplasm of renal tubular epithelial cells and no remarkable changes of immunoreactivities were observed using immunohistochemical detection after Cd treatment. The results reveal that Cd can trigger apoptosis, while it cannot activate caspase-8 in purse red common carp.

Cadmium induces liver cell apoptosis through caspase-3A activation in purse red common carp (Cyprinus carpio).

Caspase-3, the essential effector caspase, plays a pivotal role during caspase-dependent apoptosis. In this study, we isolated and characterized caspase-3A gene from common carp. The common carp caspase-3A comprising 273 amino acids showed 71.8% sequence similarity and 59.3% sequence identity to human caspase-3. It exhibited an evolutionarily conserved structure of mammalian caspase-3 genes, including a pro-domain, a large subunit, a small subunit and other motifs such as the pentapeptide active-site motif (QACRG) and the putative cleavage sites at the aspartic acids. Phylogenetic analysis demonstrated that common carp caspase-3A formed a clade with cyprinid fish caspase-3. To assess whether caspase-3A is involved in cadmium (Cd)-induced cell apoptosis in common carp, a Cd exposure experiment was performed. TUNEL analysis showed that Cd triggered liver cell apoptosis; caspase-3A activity was markedly increased; its proenzyme level was significantly decreased, and the levels of its cleaved forms were markedly increased. However, real-time quantitative PCR analysis revealed that the mRNA transcript level of caspase-3A was not significantly elevated. Immunoreactivities were observed in the cytoplasm of hepatocytes by immunohistochemical detection. The findings indicates that Cd can trigger liver cell apoptosis through the activation of caspase-3A. Caspase-3A may play an essential role in Cd-induced apoptosis.

[Monocyte chemotactic protein-1 gene polymorphism and monocyte chemotactic protein-1 expression in Chongqing Han children with tuberculosis].

OBJECTIVE: The aims of this study were to evaluate whether the presence of -2518A/G polymorphism in the distal regulatory region of the monocyte chemotactic protein-1 (MCP-1) was associated with tuberculosis (TB) in Chongqing Han population and to find whether it has a significant impact on the pediatric patient. METHOD: One hundred children [ < or = 15 years old, mean age (7.3+/-4.6) years, 53 male, 47 female] and one hundred adults [51 male, 49 female, age (44.6+/-13.5) years with TB] and 200 healthy controls of comparable age were screened for genotype by PCR-sequence-specific primer (SSP) method. MCP-1 levels in the sera were detected by ELISA. RESULT: (1) TB patients and controls showed different single nucleotide polymorphism (SNP) distribution patterns (58%, 36%). MCP-1 alleles -2518G was associated with increased TB susceptibility (P<0.01). (2) The -2518 GG genotypes was associated with increased TB susceptibility (32% in TB patients and 13% in non-TB controls respectively, P<0.01). (3) The odds of developing TB in genotypes GG were higher than those in homozygous AA, and the risk was higher in children than in adult (7.0-fold in children and 5.1-fold in adults, respectively). (4) Cases of homozygous GG had the highest plasma levels of MCP-1, which increased the likelihood of developing TB. Furthermore, higher levels were observed in children than in adults. CONCLUSION: These findings suggest that persons bearing the MCP-1 genotype GG produce high concentrations of MCP-1, which increases the risk of active TB infection in Chongqing Han people. These findings are more significant in child patients than in adult patients with TB.