GLRaV-2 protein p24 suppresses host defenses by interaction with a RAV transcription factor from grapevine.

Grapevine leafroll-associated virus 2 (GLRaV-2) is a prevalent virus associated with grapevine leafroll disease, but the molecular mechanism underlying GLRaV-2 infection is largely unclear. Here, we report that 24-kDa protein (p24), an RNA-silencing suppressor (RSS) encoded by GLRaV-2, promotes GLRaV-2 accumulation via interaction with the B3 DNA-binding domain of grapevine (Vitis vinifera) RELATED TO ABSCISIC ACID INSENSITIVE3/VIVIPAROUS1 (VvRAV1), a transcription factor belonging to the APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) superfamily. Salicylic acid-inducible VvRAV1 positively regulates the grapevine pathogenesis-related protein 1 (VvPR1) gene by directly binding its promoter, indicating that VvRAV1 may function in the regulation of host basal defense responses. p24 hijacks VvRAV1 to the cytoplasm and employs the protein to sequester 21-nt double-stranded siRNA together, thereby enhancing its own RSS activity. Moreover, p24 enters the nucleus via interaction with VvRAV1 and weakens the latter's binding affinity to the VvPR1 promoter, leading to decreased expression of VvPR1. Our results provide a mechanism by which a viral RSS interferes with both the antiviral RNA silencing and the AP2/ERF-mediated defense responses via the targeting of one specific host factor.

[Sudden unexpected postnatal collapse in a neonate].

A healthy full-term female neonate, aged 3 days and born by vaginal delivery (with a 1-minute Apgar score of 10 and a 5-minute Apgar score of 10), had unexpected cardiac and respiratory arrests in the early morning on day 3 after birth and recovered to spontaneous breathing and heartbeat after a 10-minute resuscitation. The child had poor response and convulsion after resuscitation. Blood gas analysis showed metabolic acidosis, and amplitude-integrated EEG showed a burst-suppression pattern. She was diagnosed with sudden unexpected postnatal collapse but improved after hypothermia and symptomatic/supportive treatment. This article reports the first case of sudden unexpected postnatal collapse in China and summarizes related risk factors, pathophysiological mechanisms, and preventive and treatment measures of this disorder.

[Bioinformatic analysis of target gene prediction and related signaling pathways of miRNA-126*].

OBJECTIVE: To bioinformatically predict and analyze target genes of miRNA-126(*), with the aim of providing certain basis for related research about target genes and regulatory mechanism in the future. METHODS: The miRNA chip technology was applied to measure expression levels of miRNA-126(*) in 3 time points (embryo 16, 19 and 21 days) of fetal lung development. Then the target genes of miRNA-126(*) were screened through miRGen2.0 database. Subsequent bioinformatic analysis of these target genes was performed by Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes Pathway analysis (KEGG Pathway analysis). RESULTS: miRNA-126(*) manifested continuously upregulated expression with the lung development (from embryo 16 to 21 days). There were 422 predicted target genes in total, and the gene set mainly located in glucuronosyltransferase activity, transferase activity (GO molecular function), multicellular organismal development, developmental process (GO biology process) and intracellular part (GO cellular component). The KEGG Pathway analysis demonstrated that the gene set mostly located in RNA degradation (signal transduction pathway) and prion diseases (disease pathway). CONCLUSIONS: The results suggest that miRNA-126(*) plays a certain role in fetal lung development and provide a basis for lung development research in the future.

Is epinephrine still the drug of choice during cardiac arrest in the emergency department of the hospital? A meta-analysis.

Epinephrine is the first-line emergency drug for cardiac arrest and anaphylactic reactions but is reported to be associated with many challenges resulting in its under- or improper utilization. Therefore, in this meta-analysis, the efficacy and safety of epinephrine as a first-line cardiac emergency drug for both out-of-hospital and in-hospital patients was assessed. Pertinent articles were searched in central databases like PubMed, Scopus, and Web of Science, using appropriate keywords as per the PRISMA guidelines. Retrospective and prospective studies were included according to the predefined PICOS criteria. RevMan and MedCalc software were used and statistical parameters such as odds ratio and risk ratio were calculated. Twelve clinical trials with a total of 208,690 cardiac arrest patients from 2000 to 2022 were included, in accordance with the chosen inclusion criteria. In the present meta-analysis, a high odds ratio (OR) value of 3.67 (95 % CI 2.32-5.81) with a tau2 value of 0.64, a chi2 value of 12,446.86, df value of 11, I2 value of 100 %, Z-value 5.53, and a p-value < 0.00001 were reported. Similarly, the risk ratio of 1.89 (95 % CI 1.47-2.43) with a tau2 value of 0.19, chi2 value of 11,530.67, df value of 11, I2 value of 100 %, Z-value of 4.95, and p-value < 0.000001. The present meta-analysis strongly prefers epinephrine injection as the first cardiac emergency drug for both out-of-hospital and in-hospital patients during cardiac arrest.

[Role of miRNA-126/miRNA-126* in the fetal lung development of rats].

OBJECTIVE: To investigate the expression and role of miRNA-126/miRNA-126(*) in the fetal lung development of rats. METHODS: Twelve pregnant Sprague-Dawley rats were randomly divided into 3 groups and the fetal rats were removed at 16, 19 and 21 days of gestation respectively. Hematoxylin and eosin staining was performed to observe lung morphology of fetal rats. Then microRNA (miRNA) microarray was used to study the expression patterns of miRNA-126/miRNA-126(*) in fetal lungs at the three time points. And miRNA-126(*) was selected for further study by real-time PCR. RESULTS: There was no evident difference in the expression of miRNA-126 among the three groups, however the expression level of miRNA-126(*) increased gradually as the fetal lung developed. The real-time PCR result further showed that expression of miRNA-126(*) increased gradually with lung development, displaying significant differences among the three groups (P<0.05). CONCLUSIONS: miRNA-126(*) may play an important role in development of the fetal lung in rats.

Elizabethkingia anophelis: An Important Emerging Cause of Neonatal Sepsis and Meningitis in China.

Elizabethkingia anophelis, originally isolated from the midgut of Anopheles gambiae in 2011, is an important cause of sepsis in adults and children and meningitis in newborns, with several reported outbreaks worldwide. Accumulating molecular biological and whole-genome sequencing (WGS) evidence suggests that E. anophelis is the major human pathogen belonging to the genus Elizabethkingia. The source of infection, routes of transmission and pathogenicity of E. anophelis are unclear and should be better understood as the bacterium is capable of causing sepsis and meningitis in newborns, with complications and high mortality rates. Here, we describe two healthy neonates who developed meningitis caused by Elizabethkingia infection. Initial conventional laboratory results revealed that the pathogen was E. meningoseptica; metagenomic findings later confirmed it as E. anophelis. We also summarize reported E. anophelis infections among newborns in China and elsewhere and describe the clinical, pathogenic and genetic characteristics of this bacillus.

Clinical features and outcomes of congenital chylothorax: a single tertiary medical center experience in China.

OBJECTIVE: Congenital chylothorax (CC) is an uncommon congenital disease. The objective of this study was to analyze the clinical features, treatment, and outcome of infants with CC in a Chinese tertiary medical center. METHODS: CC was defined as a non-traumatic pleural effusion with ≥ 80% lymphocytes detected before birth or within 28 days after birth. Clinical data were collected in CC infants discharged from June 2017 to March 2021. RESULTS: A total of 24 CC infants were discharged during the study period, accounting for 67% of congenital pleural effusions. The median gestational age at birth was 36+4 weeks (range 29+5-41 weeks) and the birth weight was 3025 g (range 1850-4250 g). Twenty-one infants were diagnosed antenatally. The median gestational age at the time of diagnosis was 30+3 weeks (range 24-36+6 weeks). Nine infants presented with hydrops fetalis; 18 were bilateral. Prenatal interventions were performed in 13 fetuses. Nine infants (38%) had birth asphyxia. Compared with the infants without hydrops fetalis, the infants with CC and hydrops fetalis had lower Apgar scores at 1 and 5 min (P < 0.05) and a lower gestational age at birth (P < 0.05). Postnatally, 17 infants required continuous pleural drainage for 10 days (range 2-30 days). Analysis of the pleural effusion showed a higher cell count, lymphocyte fraction, and protein content after enteral feeding (P < 0.05). Fifteen infants required mechanical ventilation; 9 did not require any respiratory support. Ten infants received a delayed feeding strategy and 17 received a medium-chain triglyceride (MCT) formula. Only 1 infant received octreotide therapy. Twenty-one infants survived and 3 died. The main cause of death was pulmonary dysplasia. The duration of hospital stay in survivors was 21.5 days (range 10-43) days. For infants with CC and hydrops fetalis, prenatal therapy shortened the duration of pleural drainage and the length of hospital stay (P < 0.05). CONCLUSION: CC is the most common cause of congenital pleural effusions. The poor prognosis is mainly associated with prematurity, hydrops fetalis, and pulmonary dysplasia. Prenatal intervention may improve the outcome of infants with hydrops fetalis.

Mannitol Improves Intraoperative Brain Relaxation in Patients With a Midline Shift Undergoing Supratentorial Tumor Surgery: A Randomized Controlled Trial.

BACKGROUND: Mannitol is widely used to reduce brain tissue swelling and improve brain relaxation during neurosurgery. However, the optimal dosage for patients with midline shift undergoing supratentorial tumor resection remains unclear. METHODS: In this randomized, controlled double-blinded study, 204 patients with preoperative midline shift who underwent elective supratentorial brain tumor surgery were equally allocated to receive placebo or 0.7, 1.0, or 1.4 g/kg mannitol infusion. The primary outcome was the proportion of satisfactory brain relaxation. RESULTS: Demographics and baseline characteristics were similar among the 4 groups. Trend analysis showed that mannitol infusion increased satisfactory brain relaxation (P<0.0001), relaxed dural tension (P<0.0001) and adequate surgical exposure (P<0.0001), and decreased the requirement for rescue therapy for brain swelling (P<0.0005), all in a dose-dependent manner. Tumor size (odds ratio [OR]: 0.99 per 1 mm, 95% confidence interval [CI]: 0.989-0.998, P=0.004), peritumoral edema classification (OR: 0.60, 95% CI: 0.37-0.97; P=0.038) as well as mannitol dose (OR: 2.81, 95% CI: 1.97-4.02, P<0.0001) were significantly associated with satisfactory brain relaxation. An increased risk of moderate to severe postoperative cerebral edema was found in the group receiving 1.4 g/kg mannitol (P=0.025) in a dose-dependent manner (P=0.018). CONCLUSIONS: An optimal mannitol infusion dosage of 1.0 g/kg is recommended to improve brain relaxation with lower risk of moderate to severe postoperative cerebral edema in patients with midline shift undergoing supratentorial tumor resections. The effect of mannitol on brain relaxation is affected by tumor size and severity of peritumoral edema, rather than by midline shift.

Expression profiles of long non-coding RNAs during fetal lung development.

With advances in neonatology, a greater percentage of premature infants now survive and consequently, diseases of lung development, including bronchopulmonary dysplasia and neonatal respiratory distress syndrome, have become more common. However, few studies have addressed the association between fetal lung development and long non-coding RNA (lncRNA). In the present study, right lung tissue samples of fetuses at different gestational ages were collected within 2 h of the induction of labor in order to observe morphological discrepancies. An Affymetrix Human GeneChip was used to identify differentially expressed lncRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed. A total of 687 lncRNAs were identified to be differentially expressed among three groups of fetal lung tissue samples corresponding to the three embryonic periods. A total of 34 significantly upregulated and 12 significantly downregulated lncRNAs (fold-change, ≥1.5; P<0.05) were detected at different time points (embryonic weeks 7-16, 16-25 and 25-28) of fetal lung development and compared with healthy tissues Expression changes in lncRNAs n340848, n387037, n336823 and ENST00000445168 were validated by reverse transcription-quantitative PCR and the results were consistent with the GeneChip results. These novel identified lncRNAs may have roles in fetal lung development and the results of the present study may lay the foundation for subsequent in-depth studies into lncRNAs in fetal lung development and subsequent clarification of the pathogenesis of neonatal pulmonary diseases.

Knockout of microRNA­26a promotes lung development and pulmonary surfactant synthesis.

Normal formation and function of the lungs are essential for the transition of the fetus to an air­breathing environment at birth. The synthesis of pulmonary surfactant (PS), which is produced by type II alveolar epithelial cells (AECIIs), is required for proper lung development. Previous in vitro studies have suggested that PS synthesis is regulated by microRNA (miR)­26a in fetal rat AECIIs. The present study explored the potential role of miR­26a in lung development and PS synthesis by using a miR­26a­1/miR­26a­2 double knockout mouse model. Hematoxylin and eosin staining and transmission electron microscopy were used to observe the morphology of fetal lungs. Reverse transcription­quantitative polymerase chain reaction and western blot analysis were performed to examine the mRNA and protein levels of surfactant­associated proteins. The results demonstrated that the lung formation in the knockout mice was more mature, and that there were more mature lamellar bodies inside AECIIs in miR­26a knockout mice at late stages of lung development. The findings further demonstrated that knockout of miR­26a increased surfactant­associated mRNA and protein expression levels. The results indicated that knockout of miR­26a promotes lung development and PS synthesis.

Pulmonary surfactant synthesis in miRNA-26a-1/miRNA-26a-2 double knockout mice generated using the CRISPR/Cas9 system.

Pulmonary surfactant (PS), which is synthesized by type II alveolar epithelial cells (AECIIs), maintains alveolar integrity by reducing surface tension. Many premature neonates who lack adequate PS are predisposed to developing respiratory distress syndrome (RDS), one of the leading causes of neonatal morbidity and mortality. PS synthesis is influenced and regulated by various factors, including microRNAs. Previous in vitro studies have shown that PS synthesis is regulated by miR-26a in fetal rat AECIIs. This study aimed to investigate the role of miR-26a in PS synthesis in vivo. To obtain a miR-26a-1/miR-26a-2 double knockout mouse model, we used the clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9 (CRISPR/Cas9) system, an important genome editing technology. Real-time PCR was performed to determine the miR-26a levels in various organs, as well as the mRNA levels of surfactant-associated proteins. Moreover, AECIIs and surfactant-associated proteins in lung tissues were analyzed by hematoxylin-eosin staining and immunohistochemistry. Homozygous offspring of miR-26a-1/miR-26a-2 double knockout mice generated using the CRISPR/Cas9 system were successfully obtained, and PS synthesis and the number of AECIIs were significantly increased in the miR-26a knockout mice. These results indicate that miR-26a plays an important role in PS synthesis in AECIIs.

Infantile Pompe disease: A case report and review of the Chinese literature.

Pompe disease, also known as glycogen storage disease type II, is caused by acid maltase deficiency, and can lead to lysosomal glycogen storage. The primal manifestations may be observed in children and adults, and also in infants. In general, the clinical spectrum in infants is more progressive and lethal than that in older patients. This case report describes the case of a newborn who was found to have cardiac hypertrophy, hepatomegaly and elevated serum enzyme levels, which was characterized by an aspartate aminotransferase level of 95 U/l, lactate dehydrogenase level of 778 U/l and creatine kinase level of 1,299 U/l. On the basis of the clinical signs and laboratory results, dried blood spots from the baby were tested to determine the acid α-glucosidase (GAA) activity, and the result confirmed that the GAA activity was only 0.10 pmol/punch/h (normal reference range, 2.88-89.02 pmol/punch/h) at pH 3.8, which was clearly lower than the normal range, leading to a diagnosis of Pompe disease. Pompe disease is incurable, and before the introduction of enzyme replacement therapy (ERT), pain relief was the main treatment. Recognizing this disease earlier and starting ERT in infants prior to the development of clinical symptoms is likely to improve the quality of life of patients.

Expression profile of plasma microRNAs in premature infants with respiratory distress syndrome.

As well-known regulators of gene expression, microRNAs (miRNAs) are important not only in cell proliferation and differentiation, but also in tumorigenesis and organ development. It has been estimated that miRNAs may be responsible for regulating the expression of almost one third of the human genome. Simultaneously, with advances in neonatal care in the clinic, an increased number of premature infants are being saved and, thus, respiratory distress syndrome (RDS) has become more common. However, previous non-miRNA studies have suggested their connection with RDS. In the present study, a miRNA microarray, including >1,891 capture probes was used to compared the expression profiles of plasma miRNAs between RDS and control groups. miRNAs, which were observed to have consistent fold-changes (fold-change ≥ 1.3) between the two groups were selected and validated using reverse transcription-quantitative polymerase chain reaction. As a result, 171 differentially expressed miRNAs were identified, including two upregulated and seven downregulated miRNAs. Of these miRNAs, four were selected as having higher fold-changes between the two groups. This is the first time, to the best of our knowledge, that these nine miRNAs have been reported in RDS. It was hypothesized that these novel miRNAs may be important in RDS, and may provide meaningful biomarkers for the diagnosis of RDS.

The expression profile of microRNAs in wistar rats with lipopolysaccharide-induced periventricular leukomalacia.

Over the recent decades, with numbers of premature infants being cured, clinical diseases on brain damage like periventricular leukomalacia (PVL) have become much more common. Meanwhile, since the discovery of first miRNA lin-4, an increasing number of important studies about this small RNA have been performed not only in the normal organ development but also in the pathogenic mechanism of diseases. However, throughout the past several years, there have been rare miRNA researches discussing the connection between the PVL and miRNA. In view of this situation, we constructed an animal model of PVL induced by lipopolysaccharide (LPS) and performed a miRNA microarray which was repeated three times to profile the expression of microRNAs (miRNAs) between two groups (PVL group versus control group). Then, miRNAs with notable fold changes (fold change >1.5) were found; some of them were further validated by real-time PCR. As a result, 104 differentially expressed miRNAs were identified using the microarray, including 64 upregulated and 40 downregulated miRNAs. Then, five miRNAs of them were selected, characterized by consistent trend in expression in all three microarrays. Among these five miRNAs (miRNA-451, miRNA-200b, miRNA-29a, miRNA-21, and miRNA-138), we subsequently selected miRNA-451 and miRNA-200b for real-time PCR because they possess the highest fold changes. Finally, the results of PCR are basically in accord with the microarray. We guess these new identified miRNAs may play an important role in the pathogenesis of PVL and may provide certain pathophysiological basis for the future research of related diseases in preterm infants.

[Clinical evaluation of the effects of morphine in mechanical ventilation of neonates].

OBJECTIVE: To evaluate the effects of morphine infusion analgesia on behavioural and neuroendocrine stress response and short term outcome in ventilated neonates. METHODS: A randomized, double-blind clinical trial was conducted between August 2010 and April 2011 at the neonatal intensive care unit of Nanjing Children's Hospital Affiliated to Nanjing Medical University. A total of 46 ventilated preterm infants (≥ 32 weeks) and term infants were divided into 2 groups at random. Twenty-two infants in test group received a loading dose (100 µg/kg) of morphine (> 1 h) followed by a continuous infusion [10 µg/(kg·h)] for (70.05 ± 29.05) h, and 24 infants in control group received 5% glucose with the same infusion rate. (1) The ventilatory parameters [respiratory rate (R), frequence (f), peak inspiratory pressure (PIP), positive end expiratory pressure (PEEP), fraction of inspired oxygen (FiO2)], mean blood pressure (MBP) and heart rate (HR) before treatment, at 30 min, 2 h, 6 h, 12 h, 24 h, 48 h after treatment between two groups were compared. (2) Pain was measured by two assessment tools [neonatal pain, agitation and sedation scale (N-PASS) and COMFORT scale] at the same periods. (3) The ventilation duration, the time from withdrawal to extubation, the total oxygen-inhaled time, the side effects and the clinical outcomes [e.g., pulmonary hemorrhage, air leak, patent ductus arteriosus (PDA), necrotizing enterocolitis (NEC), intraventricular hemorrhage (IVH)] between two groups were compared. RESULTS: (1) There were no significant differences in the different ventilatory parameters before and after treatment between two groups at different periods (P > 0.05). There was no significant difference in the average blood pressure of two groups at different periods, but the heart rate reduced at 24 - 48 h after treatment in test group with significant difference as compared to control group (t = -2.152 and -2.513, P < 0.05). (2) The N-PASS score and COMFORT score in test group were lower than that in control group at different time points 2 h after treatment (P < 0.05), especially 12 h after treatment (P < 0.01). (3) There were no significant differences in the ventilation duration, the time from withdrawal to extubation and the total oxygen time between two groups, and also in side effects, the incidence of IVH, white matter damage and the clinical outcomes. CONCLUSION: Continuous infusion of morphine could relieve pain in ventilated neonates, reduce the stress response and promote the human-machine coordination, but the medication did not show any effects on neurobehavioral development and short term outcome.