Since the outbreak of novel coronavirus infection pneumonia in Wuhan City, China, in late 2019, such cases have been gradually reported in other parts of China and abroad. Children have become susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) because of their immature immune function. As the outbreak has progressed, more cases of novel coronavirus infection/pneumonia in children have been reported. Compared with adults, the impact of SARS-CoV-2 infection in children is less severe, with a lower incidence and susceptibility in children, which results in fewer children being tested, thereby underestimating the actual number of infections. Therefore, strengthening the diagnosis of the disease is particularly important for children, and early and clear diagnosis can determine treatment strategies and reduce the harm caused by the disease to children. According to the Novel Coronavirus Infection Pneumonia Diagnosis and Treatment Standards ( trial version 7 ) issued by National Health Committee and the latest diagnosis and treatment strategies for novel coronavirus infection pneumonia in children, this review summarizes current strategies on diagnosis and treatment of SARS-CoV-2 infection in children.

AIMS:The aim of this study was to investigate the molecular mechanism underlying preterm white matter injury (WMI) via the identification and functional analysis of differentially expressed long non-coding RNAs (lncRNAs) and mRNAs.MAIN METHODS:A neonatal rat model of preterm WMI was established by ligating the common carotid artery and hypoxia induction. RNA sequencing was performed to analyze gene expression profiles of brain samples. Gene Ontology (GO) and Kyoto Encyclopedia of Genes (KEGG) analyses were performed to evaluate functions of target mRNAs. A co-expression network was generated to explore regulatory mechanisms.KEY FINDINGS:In total, 210 lncRNAs and 619 mRNAs were differentially expressed between the preterm WMI group and the sham group. Based on GO and KEGG analyses, enriched pathways included the apoptotic signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway, natural killer cell-mediated cytotoxicity pathway, and the autophagy pathway.SIGNIFICANCE:Differentially expressed lncRNAs and mRNAs in the brain tissues of preterm WMI model were identified, and the biological processes were closely associated with the development of preterm WMI, thus being considered potential targets for future studies.

Necrotizing enterocolitis (NEC) is one of the most common and devastating gastrointestinal diseases in preterm newborns, and its underlying mechanisms remain unclear. Non-coding RNAs (ncRNAs) play critical roles in intestinal diseases; however, little is known about their roles in the development of NEC. To gain a deeper understanding of the pathophysiological mechanism of NEC, long non-coding RNAs (lncRNAs), microRNAs (miRNAs) and mRNAs were detected in an NEC rat model. In total, 1820 lncRNAs, 118 miRNAs and 929 mRNAs were differentially expressed in NEC group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these molecules were enriched in apoptosis, autophagic cell death, TLR4 signaling pathway, Notch signaling pathway, and mTOR signaling pathway. These pathways are thought to be closely associated with NEC. Furthermore, a lncRNA-miRNA interaction network was constructed, and four of the novel, differentially expressed lncRNAs with large changes were randomly verified using quantitative polymerase chain reaction (qPCR). The GO and KEGG pathway analysis of these four lncRNAs showed that they were associated with the negative regulation of TLR4 signaling pathway and Notch signaling pathway. In conclusion, our study revealed that these differentially expressed lncRNAs may participate in the development of NEC via interactions with miRNAs and may serve as possible biomarkers and target genes in NEC.

Idiopathic nephrotic syndrome (INS) is a disease involving injury to podocytes in the glomerular filtration barrier, and its specific causes have not been elucidated. Transfer RNA-derived fragments (tRFs), products of precise tRNA cleavage, have been indicated to play critical roles in various diseases. Currently, there is no relevant research on the role of tRFs in INS. This study intends to explore the changes in and importance of tRFs during podocyte injury in vitro and to further analyze the potential mechanism of INS. Differentially expressed tRFs in the adriamycin-treated group were identified by high-throughput sequencing and further verified by quantitative RT-PCR. In total, 203 tRFs with significant differential expression were identified, namely, 102 upregulated tRFs and 101 downregulated tRFs (q<0.05, ∣log2FC∣≥2). In particular, AS-tDR-008924, AS-tDR-011690, tDR-003634, AS-tDR-013354, tDR-011031, AS-tDR-001008, and AS-tDR-007319 were predicted to be involved in podocyte injury by targeting the Gpr, Wnt, Rac1, and other genes. Furthermore, gene ontology analysis showed that these differential tRFs were strongly associated with podocyte injury processes such as protein binding, cell adhesion, synapses, the actin cytoskeleton, and insulin-activate receptor activity. KEGG pathway analysis predicted that they participated in the PI3K-Akt signaling pathway, Wnt signaling pathway, and Ras signaling pathway. It was reported that these pathways contribute to podocyte injury. In conclusion, our study revealed that changes in the expression levels of tRFs might be involved in INS. Seven of the differentially expressed tRFs might play important roles in the process of podocyte injury and are worthy of further study.

Background and aim Human milk has potential protective effects against bronchopulmonary dysplasia (BPD). However, studies on the association between the dose of human milk and BPD in China are limited. This study aimed to evaluate the dose-dependent effects of human milk on BPD and other neonatal morbidities in very low birth weight (VLBW) infants. Methods This retrospective cohort study of preterm infants was conducted on preterm infants of gestational age ≤ 34 weeks and birth weight < 1500 g admitted to the multicenter clinical research database for breastfeeding quality improvement in Jiangsu province. The multivariate analysis was performed to compare the effect outcomes of daily graded doses [1–24 mL/(kg · day), 25–49 mL/(kg · day), and ≥ 50 mL/(kg · day) of body weight] of human milk on neonatal outcomes throughout the first 4 weeks of life versus a reference group receiving no human milk. The models were adjusted for potential confounding variables. Results Of 964 included infants, 279 (28.9%) received exclusive preterm formula, 128 (13.3%) received 1–24 ml/(kg · day), 139 (14.4%) received 25–49 ml/(kg · day), and 418 (43.4%) received ≥50 ml/(kg · day) human milk for the first 4 weeks of life. Compared with infants receiving exclusive formula, those receiving the highest volume of human milk daily [≥50 mL/(kg · day)] had lower incidences of BPD [27.5% in ≥50 mL/(kg · day) vs 40.1% in 0 mL/(kg · day) human milk, P = 0.001)], moderate and severe BPD [8.9% in ≥50 mL/(kg · day) vs 16.1% in 0 mL/(kg · day), P = 0.004], necrotizing enterocolitis [NEC; 3.8% in ≥50 mL/(kg · day) vs 10.8% in 0 mL/(kg · day), P = 0.001], late-onset sepsis [LOS; 9.3% in ≥50 mL/(kg · day) vs 19.7% in 0 mL/(kg · day), P <0.01], and extrauterine growth retardation [EUGR; 38.5% in ≥50 mL/(kg · day) vs 57.6% in 0 mL/(kg · day), P <0.01)]. The logistic regression indicated that those receiving ≥50 ml/kg · day human milk had lower odds of BPD [adjusted odds ratio (AOR) 0.453; 95% confidence interval (CI): 0.309, 0.666], moderate and severe BPD (AOR 0.430; 95% CI: 0.249, 0.742), NEC (AOR 0.314; 95% CI: 0.162, 0. 607), LOS (AOR 0.420; 95% CI: 0.263, 0.673), and EUGR (AOR 0.685; 95% CI: 0.479, 0.979). Conclusions A daily threshold amount of ≥50 ml/(kg · day) human milk in the first 4 weeks of life was associated with lower incidence of BPD as well as NEC, LOS, and EUGR in VLBW infants. Trial registration ClinicalTrials.gov Identifier: NCT03453502 . Registration date: March 5, 2018. This study was retrospectively registered.

Background Human breast milk has potential protective effects against bronchopulmonary dysplasia (BPD). However, limited multicenter research has been reported on the association between the dose of maternal milk and BPD in China. In this study, we aimed to evaluate the dose effects of maternal milk on BPD and other neonatal morbidities in very low birth weight (VLBW) infants. Methods We conducted a retrospective cohort study of preterm infants of gestational age ≤ 34 weeks and birth weight < 1500 g admitted to the multicenter clinical research database for breastfeeding quality improvement in Jiangsu province. Multivariate analysis was performed to compare the effect on neonatal outcomes of daily graded doses of maternal milk throughout the first 4 weeks of life versus a reference group receiving no maternal milk. Models were adjusted for potential confounding variables. Results Of 964 included infants, 279 (28.9%) received exclusive preterm formula, another 128 (13.3%) received 1–24 ml/kg, 139 (14.4%) received 25–49 ml/kg, and 418 (43.4%) received ≥ 50 ml/kg maternal milk for the first 4 weeks of life. Compared with infants receiving exclusive formula, those receiving the highest volume of maternal milk daily (≥ 50 ml/kg) had lower incidences of BPD (27.5% in ≥ 50 ml/kg maternal milk vs. 40.1% in formula), moderate and severe BPD (8.9% in ≥ 50 ml/kg maternal milk vs. 16.1% in formula), necrotizing enterocolitis (NEC; 3.8% in ≥ 50 ml/kg maternal milk vs. 10.8% in formula), late-onset sepsis (LOS; 9.3% in ≥ 50 ml/kg maternal milk vs. 19.7% in formula), and extrauterine growth retardation (EUGR; 38.5% in ≥ 50 ml/kg maternal milk vs. 57.6% in formula). Logistic regression indicated that those receiving ≥ 50 ml/kg/day maternal milk had lower odds of BPD (adjusted odds ratio [AOR] 0.453; 95% confidence interval [CI]: 0.309, 0.666), moderate and severe BPD (AOR 0.430; 95% CI: 0.249, 0.742), NEC (AOR 0.314; 95% CI: 0.162, 0. 607), LOS (AOR 0.420; 95% CI: 0.263, 0.673), and EUGR (AOR 0.685; 95% CI: 0.479, 0.979). Conclusions A daily threshold amount of ≥ 50 ml/kg maternal milk in the first 4 weeks of life was associated with lower incidence of BPD as well as NEC, LOS, and EUGR in VLBW infants. Trial registration: ClinicalTrials.gov Identifier: NCT03453502

Oxidative stress and exaggerated inflammatory response play crucial roles in necrotizing enterocolitis (NEC). Although Saccharomyces boulardii (S. boulardii) participates in many different biological activities, it remains unknown whether S. boulardii inhibits cell death induced by NEC. Furthermore, mechanisms against oxidative stress levels have not yet been elucidated. The present study established an experimental mouse model of NEC obtained after artificial feeding + hypoxia/re-oxygenation cooling stimulation + lipopolysaccharide for 3 days. Hematoxylin and eosin (HE) staining and morphologic studies were performed to define the model. After S. boulardii treatment, weights and survivor numbers were significantly increased, while intestinal tissue damage was reduced. Necroptosis and apoptosis were inhibited by S. boulardii. Protein and mRNA expression levels of caspase-3, caspase-8, and RIP3 further confirmed the protection of S. boulardii. Furthermore, S. boulardii increased super oxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) levels, while promoting clearing of reactive oxygen species (ROS) and malondialdehyde (M DA). The current study provides the first evidence that S. boulardii exerts anti-apoptosis and necroptosis effects in NEC intestinal tissues through increasing ROS consumption.

AimsThe purpose of this study was to investigate the pathophysiology and discover novel predictors of neonatal respiratory distress syndrome (NRDS) from a peptidomics perspective.Main methodsComparative profiling of umbilical cord blood from NRDS and control patients was performed by liquid chromatography tandem mass spectrometry technology. The underlying biological functions of the differentially expressed peptides (DEPs) were predicted by Gene Ontology (GO) and KEGG pathway analyses. The interactions of DEPs and their precursor proteins were explored by ingenuity pathway analysis (IPA). The sources and stability of DEPs were determined by online databases, including UniProt, SMART and ProtParam tool.Key findingsA total of 251 DEPs were identified, of which 139 peptides were upregulated, and 112 peptides were downregulated (fold change ≥2.0, P < 0.05). These DEPs were predicted to be associated with respiratory failure, atelectasis, and morphogenesis of endothelial cells. These processes indicated that DEPs may play a role in NRDS. Among them, eleven stable DEPs might be used as preclinical biomarkers.SignificanceOur findings improve our understanding of NRDS and facilitate the discovery of candidate diagnostic biomarkers for NRDS from the perspective of peptidomics.

microRNAs (miRNAs) have been demonstrated to contribute to tumor progression and metastasis, and have been proposed to be key regulators of diverse biological processes. In this study, we report that miR-4295 is deregulated in bladder cancer tissues and cell lines. To characterize the role of miR-4295 in bladder cancer cells, we performed functional assays. The overexpression of miR-4295 significantly promoted bladder cancer cell proliferation, colony formation, and migration. Moreover, its downregulation induced cell cycle arrest and apoptosis of bladder cancer cells. Furthermore, a luciferase reporter assay and rescue experiment indicated that miR-4295 directly targets BTG1 by binding its 3'UTR. In conclusion, these results demonstrate that miR-4295 acts as an oncogene and may be a potential biomarker for bladder cancer diagnosis and treatment.

The aim of this research is to explore the effect of miR-200b-3p targeting DNMT3A on the proliferation and apoptosis of osteoarthritis (OA) cartilage cells. Quantitative RT-PCR was performed to analyse the expression of miR-200b-3p, DNMT3A, MMP1, MMP3, MMP9, MMP13 and COL II in normal and OA cartilage tissues. The dual-luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR-200b-3p and DNMT3A. We also constructed eukaryotic expression vector to overexpress miR-200b-3p and DNMT3A. We detected the expression level of MMPs and COL II in stable transfected cartilage cells using RT-PCR and Western blot. Cell proliferation and apoptosis were evaluated using the MTS, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR-200b-3p targeting DNMT3A on the proliferation and apoptosis of OA cartilage cells. The results of RT-PCR indicated that both miR-200b-3p and COL II were down-regulated in OA cartilage tissues, while the expression of DNMT3A and MMPs was up-regulated in OA cartilage tissues. The expressions of DNMT3A, MMPs and COL II detected by Western blot showed the same trend of the results of RT-PCR. The dual-luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR-200b-3p and DNMT3A. In overexpressed miR-200b-3p cartilage cells, DNMT3A and MMPs were significantly down-regulated, COL II was significantly up-regulated, cell viability was enhanced and apoptosis rate was decreased (P < 0.05). In overexpressed DNM3T cartilage cells, MMPs were significantly up-regulated, COL II was significantly down-regulated, cell viability was weakened and apoptosis rate was increased (P < 0.05). MiR-200b-3p inhibited the secretion of MMPs, promoted the synthesis of COL II and enhanced the growth and proliferation of OA cartilage cells through inhibiting the expression of DNMT3A.

As a member of membrane-spanning 4A family, transmembrane protein 176A (TMEM176A) has been recently reported to be overexpressed in various cancers. However, the biological functions of TMEM176A in nonsmall cell lung cancer (NSCLC) remain unclear. In this study, we investigated the expression of TMEM176A in publicly available Oncomine database and found it was upregulated in lung cancer tissues compared with normal lung tissues. Similar results were also found in fresh primary NSCLC tumor tissues and corresponding normal tissues using Western blot analysis.Then the expression of TMEM176A was efficiently silenced in human NSCLC cell lines, A549 and 95D by RNA interference. Knockdown of TMEM176A obviously inhibited cell growth and colony formation, induced cell arrest at G0/G1 phase and promoted cell apoptosis in NSCLC cells, as determined by MTT, clonogenic assay and flow cytometry assays. Mechanistically, inhibition of TMEM176A significantly reduced the expression of CDK4 and Cyclin D1, but increased the expression of caspase-3, PARP cleavage and bad. In conclusion, these results firstly demonstrated that TMEM176A might serve as a potential oncogene that involved in the cell proliferation of NSCLC.

Objective: To investigate the effects and mechanisms of p38 signaling pathway in pentagastrin-regulated cell proliferation of colorectal carcinoma cell line HT-29. Methods: HT-29 cell line of colorectal carcinoma was in vitro incubated and divided into the control group, pentagastrin group, proglumide group, and pentagastrin + proglumide group. MTT reduction assay was performed to detect the proliferation status of HT-29 cell line and determine the optimal dosage of pentagastrin and proglumide. Annexin V-fluorescein isothiocyanate flow cytometry was used to detect the proliferation index (PI) and apoptosis rate (AR) of HT-29 cells. Reverse transcriptase polymerase chain reaction was performed to detect the mRNA expression of the pentagastrin receptor/cholecystokinin-B receptor (CCK-BR) and p38. The protein and phosphorylation levels of p38 were estimated by western blotting. Results: RT-PCR detection showed that CCK-BR mRNA was expressed in the HT-29 cell line. Pentagatrin improved HT-29 cell proliferation in dosage of 6.25 - 100 mg/L, and the optimal dosage of pentagastrin was 25.0 mg/L. Proglumide had no significant effect on the proliferation of HT-29 cells, but significantly inhibited the proliferation of HT-29 cells stimulated by pentagastrin when the dosage of proglumide was 8.0 - 128.0 mg/L, and the optimal dosage was 32.0 mg/L. The AR in the pentagastrin group was significantly lower than that in the control group and in the pentagastrin + proglumide group. The PI in the pentagastrin group was significantly higher than that in the control group and in the pentagastrin + proglumide group. P38 phosphorylation level in the pentagastrin group was significantly lower than that in the control group, and in the pentagastrin + proglumide group. There were no significant differences in the mRNA and protein expression of p38 in the control, pentagastrin, proglumide and pentagastrin + proglumide groups. Conclusion: Pentagastrin can improve proliferation of the CRC cell line HT-29 and inhibit apoptosis via the p38 signal transduction pathway. This mechanism may be associated with suppressed p38 protein phosphorylation level due to inhibition of proglumide, a gastrin receptor antagonist.