Novel Pathogenic Mutation of PNPLA1 Identified in Autosomal Recessive Congenital Ichthyosis: A Case Report.

Autosomal recessive congenital ichthyosis (ARCI) is characterized by being born as collodion babies, hyperkeratosis, and skin scaling. We described a collodion baby at birth with mild ectropion, eclabium, and syndactyly. Whole exome sequencing showed a compound heterozygous variant c.[56C>A], p.(Ser19X) and c.[100G>A], p.(Ala34Thr) in the PNPLA1 gene [NM_001145717; exon 1]. The protein encoded by PNPLA1 acts as a unique transacylase that specifically transfers linoleic acid from triglyceride to ω-hydroxy fatty acid in ceramide, thus giving rise to ω-O-acylceramide, a particular class of sphingolipids that is essential for skin barrier function. The variant was located in the patatin core domain of PNPLA1 and resulted in a truncated protein which could disrupt the function of the protein. This case report highlights a novel compound heterozygous mutation in PNPLA1 identified in a Chinese child.

Novel Pathogenic Mutation of PNPLA1 Identified in Autosomal Recessive Congenital Ichthyosis: A Case Report / 中国医学科学杂志(英文版)

Autosomal recessive congenital ichthyosis (ARCI) is characterized by being born as collodion babies, hyperkeratosis, and skin scaling. We described a collodion baby at birth with mild ectropion, eclabium, and syndactyly. Whole exome sequencing showed a compound heterozygous variant c.[56C>A], p.(Ser19X) and c.[100G>A], p.(Ala34Thr) in the PNPLA1 gene [NM_001145717; exon 1]. The protein encoded by PNPLA1 acts as a unique transacylase that specifically transfers linoleic acid from triglyceride to ω-hydroxy fatty acid in ceramide, thus giving rise to ω-O-acylceramide, a particular class of sphingolipids that is essential for skin barrier function. The variant was located in the patatin core domain of PNPLA1 and resulted in a truncated protein which could disrupt the function of the protein. This case report highlights a novel compound heterozygous mutation in PNPLA1 identified in a Chinese child.

Silencing of long non­coding antisense RNA brain­derived neurotrophic factor attenuates hypoxia/ischemia­induced neonatal brain injury.

Hypoxic/ischemic (HI) brain damage (HIBD) is a major cause of acute neonatal brain injury, leading to high mortality and serious neurological deficits. The antisense RNA of brain­derived neurotrophic factor (BDNF­AS) is transcribed from the opposite strand of the BDNF gene. The aim of the present study was to investigate the role of BDNF­AS in HI­induced neuronal cell injury in vivo and in vitro. Reverse transcription­quantitative PCR (RT­qPCR) assays indicated that BDNF­AS expression was significantly upregulated in HI­injured neonatal brains and hippocampal neurons. However, BDNF expression was downregulated in HI­injured neonatal brains and hippocampal neurons. Cell Counting Kit­8 assays, Hoechst staining, calcein­AM/PI staining, immunostaining, water maze tests and rotarod tests demonstrated that BDNF­AS silencing protected against hypoxia­induced primary hippocampal neuron injury in vitro and HI­induced brain injury in vivo. Mechanistically, RT­qPCR assays and western blotting indicated that BDNF­AS silencing led to increased expression of BDNF and activated the BDNF­mediated signaling pathway, as demonstrated by increased expression levels of BDNF, phosphorylated­Akt and phosphorylated­tropomyosin receptor kinase B. Collectively, the present study provides important insights into the pathogenesis of HIBD, and it was indicated that BDNF­AS silencing may be a promising approach for the treatment of neonatal HIBD.

[Safety and efficacy of caffeine use started at different time in preterm infants: a multicenter study in Jiangsu Province, China].

OBJECTIVE: To study the efficacy and safety of caffeine used in the early (≤72 hours after birth) and late (>72 hours after birth) stage in preterm infants with a gestational age of ≤31 weeks. METHODS: A retrospective analysis was performed for 640 preterm infants (with a gestational age of ≤31 weeks) who were admitted to the neonatal intensive care unit of eight hospitals in Jiangsu Province, China. Of the 640 preterm infants, 510 were given caffeine in the early stage (≤72 hours after birth; early use group) and 130 were given caffeine in the late stage (>72 hours after birth; late use group). The clinical data were compared between the two groups. RESULTS: There were no significant differences in birth weight, Apgar score, sex, gestational age, and age on admission between the two groups (P>0.05). Compared with the late use group, the early use group had a significantly younger age at the beginning and withdrawal of caffeine treatment (P<0.05) and a significantly shorter duration of caffeine treatment (P<0.05). There was no significant difference in respiratory support on admission between the two groups (P>0.05). Compared with the late use group, the early use group had significantly lower incidence rate of apnea (P<0.05) and significantly shorter oxygen supply time and length of hospital stay (P<0.05). There were no significant differences between the two groups in the incidence rates of neonatal intracranial hemorrhage, periventricular leukomalacia, necrotizing enterocolitis, retinopathy of prematurity, and patent ductus arteriosus at discharge and NBNA score at the corrected gestational age of 40 weeks (P>0.05). However, significant differences were found in the incidence of bronchopulmonary dysplasia and the rate of home oxygen therapy, but there was no significant difference in the mortality rate between the two groups (P>0.05). CONCLUSIONS: Early use of caffeine can shorten the duration of caffeine treatment, oxygen supply time, and length of hospital stay, with little adverse effect, in preterm infants with a gestational age of ≤31 weeks.

Long non-coding RNA Snhg3 protects against hypoxia/ischemia-induced neonatal brain injury.

Hypoxic-ischemic brain damage (HIBD) is a major cause of morbidity and mortality in the preterm and term infant. However, the precise mechanism of HIBD remains largely elusive. As a newly discovered long non-coding RNA, small nucleolar RNA host gene 3 (Snhg3) has shown its important roles in cell apoptosis, proliferation, and disease development. In this study, we determined the role of Snhg3 in the pathogenesis of HIBD. Snhg3 expression was significantly down-regulated in the neonatal brain and primary hippocampal cells response to hypoxic/ischemic stress. Snhg3 overexpression protected against hypoxic/ischemic-induced brain injury in vivo and hippocampal cell injury in vitro. Snhg3 acted as the sponge of miR-196 in the hippocampal cells by regulating the expression of miR-196 target genes, XIAP and CAAP1. Moreover, Snhg3 overexpression decreased brain infarct size and ameliorated hypoxic-ischemic neonatal brain damage. This study suggests that Snhg3 is a potential target for the treatment of HIBD.

GAS5 silencing protects against hypoxia/ischemia-induced neonatal brain injury.

Hypoxic/ischemic brain damage (HIBD) leads to high neonatal mortality and severe neurologic morbidity. However, the molecular mechanism of HIBD in the neonatal infant is still elusive. Long non-coding RNAs are shown as important regulators of brain development and many neurological diseases. Here, we determined the role of long noncoding RNA-GAS5 in HIBD. GAS5 expression was significantly up-regulated in hypoxic/ischemic-injured neonatal brain and hippocampal neurons. GAS5 silencing protected against hypoxic/ischemic-induced brain injury in vivo and primary hippocampal neuron injury in vitro. Mechanistically, GAS5 regulated hippocampal neuron function by sponging miR-23a. Intracerebroventricular injection of GAS5 shRNA significantly decreased brain GAS5 expression, reduced brain infarct size, and improved neurological function recovery. Collectively, this study suggests a promising therapeutic approach of GAS5 inhibition in the treatment of neonatal HIBD.

Application effect of extensively hydrolyzed milk protein formula and follow-up in preterm children with a gestational age of less than 34 weeks: study protocol for a randomized controlled trial.

BACKGROUND: The average incidence of preterm birth in the world is up to 11.1 %, and deaths of preterm children account for more than 50 % of neonatal deaths. Gastrointestinal function of preterm children with a gestational age less than 34 weeks is immaturely developed. For preterm children who can only be fed with formula due to their mothers' sickness, choosing a suitable formula can not only meet the high nutritional needs of preterm children, but also solve their low gastrointestinal tolerability, and is thus very important. METHODS/DESIGN: The study is a prospective, randomized, single-blind and controlled clinical trial. Preterm children with a gestational age less than 34 weeks meeting the inclusion criteria who cannot be breastfed will be included. To demonstrate the application effect of extensively hydrolyzed milk protein formula on the target population, preterm children will be randomized into two groups, 185 subjects in each group. The observation group will be fed with extensively hydrolyzed milk protein (100 % whey protein) formula, while the control group will be fed with preterm children's formula until the children are discharged from the neonatal intensive care unit (NICU). All the formula involved in this study will be from Dumex. After discharge, both groups will be uniformly fed with formula for 0 to 6-month-old infants. For statistical analysis, a chi-square test and Student's t test will be applied using SAS 9.4. DISCUSSION: This will be the first randomized controlled clinical study with long-term observation of the growth and development of preterm children during the NICU stay and at 3-month follow-up after discharge from the NICU. Results from this study will be used to determine whether the extensively hydrolyzed formula is more suitable for the low gastrointestinal tolerability of preterm children, and also whether feeding preterm children who are fed with such formula during the NICU stay with ordinary infant formula after discharge from the NICU would affect the normal growth and development of preterm children in the early stage of their lives. TRIAL REGISTRATION: This study was registered with the Chinese Clinical Trial Registry ( http://www.chictr.org.cn/ ) with number ChiCTR-IOR-14005696 , on December 22, 2014.

[Effects of maternal deficiency of folic acid during pregnancy on pulmonary development and SP-A expression in newborn rats].

OBJECTIVE: This study examined the effects of maternal deficiency of folic acid during pregnancy on pulmonary development and protein A (SP-A) expression in newborn rats in order to explore the possible mechanism of lung developmental disorders. METHODS: Thirty-six adult Sprague-Dawley female rats were randomly assigned into two groups: control and study (n=18). The study and the control groups were fed with fodder containing folic acid or not respectively. Two weeks later, the female rats in the two groups copulated with normal male rats. Newborn rats were sacrificed at 1, 7 and 14 days after birth (8 pups at each time point). Lung sections were stained with hematoxylin and eosin for histological examination. SP-A expression of protein and mRNA were determined by immunohistochemistry and real-time quantitative RT-PCR, respectively. RESULTS: The newborn rats from the study group showed damaged lung tissue structures. The mean optical density of type II cells with positive expression of SP-A decreased significantly from 1 to 14 days in newborn rats of the study group compared with the control newborn rats (P<0.05). The real-time quantitative RT-PCR showed that the expression of lung SP-A mRNA also decreased significantly from 1 to 14 days in newborn rats of the study group compared with control newborn rats (P<0.05). CONCLUSIONS: Maternal deficiency of folic acid during pregnancy can decrease the expression of SP-A in lung tissues of newborn rats, which might lead to the disorder of lung development maturation.

[Expression of mRNA of vascular endothelial growth factor in a rat model of hyperoxia-induced retinopathy].

OBJECTIVE: To examine the expression of mRNA of vascular endothelial growth factor (VEGF) in a rat model of hyperoxia-induced retinopathy and to investigate the role of VEGF in the process of neovascularization in retinopathy. METHODS: One hundred fifty one-day-old neonatal Sprague-Dawley rats were randomly divided into hyperoxia-induced retinopathy and normal control groups. The rats in the retinopathy group were exposed to (80 +/- 2)% oxygen for 7 days and then replaced by room air. The rats in the control group were exposed to room air all the time of the experiment. The morphologic changes of retinal vessels were estimated by observing the vascular pattern in adenosine diphosphate ase (ADPase) stained retina flat mounts. The newborn vessels were quantified by haematoxylin and eosin staining. Reversal transcription-polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA expression. RESULTS: In the retinopathy group at 7 days of age, most of central radial vessels became constricted and blocked, and central perfusion decreased obviously. After switching to room air exposure for 7 days (14 days of age), noticeable retinal neovascularization appeared. The expression of VEGF mRNA in the retinopathy group at 7 days of hyperoxia exposure was noticeably lower than in the control group, and increased gradually after switching to room air exposure. At 9 and 14 days of age, the expression of VEGF mRNA in the retinopathy group was noticeably higher than in the control group. The expression of retinal VEGF mRNA in the retinopathy group increased before neovascularization occurred, and decreased with regression of new vessels. CONCLUSIONS: Hyperoxia exposure may decrease the transcription of VEGF mRNA and the growth of retinal blood vessels. The relative hypoxia after hyperxia withdrawal can up-regulate the transcription of VEGF mRNA, resulting in a significant retinal neovascularization. The abnormal expression of VEGF in the retina may play an important role in the development of neovascularization in retinopathy.

[Study on effect of radix astragali on injury of cerebral cortex in neonatal rats after hypoxia/ischemia brain damage].

OBJECTIVE: To study the mechanism of injury of cortical nerve cell in the newborn with hypoxia/ischemia brain damage (HIBD), and the neuroprotective effect of Radix Astragali (RA). METHODS: Neonatal HIBD model rats were established and divided into the sham group, the model group and the RA group. Brain of rats obtained at different time points after HIBD to conduct histopathological examination, neuron death rate count, as well as determination of caspase-3 (cysteinyl aspartate-specific proteinase-3) protein mRNA expression in cerebral cortex by immunohistochemistry, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) respectively. RESULTS: In the model group, caspase-3 mRNA and protein showed an increase at 6 hrs, reached the peak at 24 hrs, and decreased at 48 hrs after HIBD, on the 5th and 7th day restored to baseline level. After being treated by RA, the neuron death rate of ligated side was obviously reduced, caspase-3 mRNA and protein expression peak value decreased by 45% (mRNA) and 40% - 43% (protein). CONCLUSION: RA shows markedly neuron protection in immature brain cortex after HIBD, which is related with the inhibition on caspase-3 expression.

[Neuroprotective effects of Astragulus membranaceus on hypoxia-ischemia brain damage in neonatal rat hippocampus].

OBJECTIVE: To study neuroprotective effects of astragulus membraneaceus on a neonatal rat hippocampus of hypoxia-ischemia brain damage (HIBD). METHOD: The neonatal hypoxia-ischemia model was established with 7-day-old rat pups. Brain injury was examined by neuron death rate in the hippocampal CA1 area. Caspase-3 (cysteinyl aspartate-specific proteinase) mRNA expression in ipsilateral hippocampal was measured by half-quantitative reverse transcription and polymerization chain reaction (RT-PCR). 90-day-old rats were used in tri-equal-arm maze to observe discrimination learning ability. Sham, model and astragulus-membraneaceus treated groups were set up. RESULT: In model group, caspase-3 mRNA showed an increase at 6h, with maximum arrivimg at 24 h - 48 h after HI. In astragulus-membraneaceus treated group, neurons death rate and caspase-3 mRNA were significantly reduced by astragulus membraneaceus, and discrimination learning ability of developed rats were improved obviously. CONCLUSION: Astragulus membraneaceus has a strong protective effect on neuronal damage in the immature rat hippocampus, which is ralated reducing caspase-3 expression.