Targeted Linked-Read Sequencing for Direct Haplotype Phasing of Parental GJB2/SLC26A4 Alleles: A Universal and Dependable Noninvasive Prenatal Diagnosis Method Applied to Autosomal Recessive Nonsyndromic Hearing Loss in At-Risk Families.

Noninvasive prenatal diagnosis (NIPD) for autosomal recessive nonsyndromic hearing loss (ARNSHL) has been rarely reported until recent years. However, the previous method could not be performed on challenging genome loci (eg, copy number variations, deletions, inversions, or gene recombinants) or on families without proband genotype. Here, this study assesses the performance of relative haplotype dosage analysis (RHDO)-based NIPD for identifying fetal genotyping in pregnancies at risk of ARNSHL. Fifty couples carrying pathogenic variants associated with ARNSHL in either GJB2 or SLC26A4 were recruited. The RHDO-based targeted linked-read sequencing combined with whole gene coverage probes was used to genotype the fetal cell-free DNA of 49 families who met the quality control standard. Fetal amniocyte samples were genotyped using invasive prenatal diagnosis (IPD) to assess the performance of NIPD. The NIPD results showed 100% (49/49) concordance with those obtained through IPD. Two families with copy number variation and recombination were also successfully identified. Sufficient specific informative single-nucleotide polymorphisms for haplotyping, as well as the fetal cell-free DNA concentration and sequencing depth, are prerequisites for RHDO-based NIPD. This method has the merits of covering the entire genes of GJB2 and SLC26A4, qualifying for copy number variation and recombination analysis with remarkable sensitivity and specificity. Therefore, it has clinical potential as an alternative to traditional IPD for ARNSHL.

Impact of Saccharomyces boulardii on jaundice in premature infants undergoing phototherapy

Abstract Objectives To evaluate the therapeutic effect of Saccharomyces boulardii supplementation on jaundice in premature infants undergoing phototherapy. Methods In this article, the authors reviewed 100 hospitalized jaundiced premature infants under 35 weeks of gestational age. All infants were assigned to a control group (n= 45) and a treatment group (n= 55) randomly. The infants in the treatment group received S. boulardii supplementation by undergoing phototherapy and the infants in the control group were only treated by phototherapy. The total serum bilirubin levels were detected before and at the end of phototherapy, and transcutaneous bilirubin levels were measured on the 1st, 4th, 8th and 15th day of treatment. The duration of jaundice resolution and phototherapy, stool frequency, and characteristics were compared after phototherapy. Results The duration of jaundice resolution and phototherapy were shortened. Total serum bilirubin level was lower than the control group at the end of phototherapy (p < 0.05). Transcutaneous bilirubin levels decreased more significantly on the 8th and 15th day of treatment (p < 0.05), while there were no significant differences on the post-treatment 1st and 4th day (p > 0.05). In addition, bowel movements including stool frequency and Bristol Stool Form Scale ratings of stools also improved after treatment. Conclusions S. boulardii in combination with phototherapy is effective and safe in reducing bilirubin levels and duration of phototherapy, accelerating jaundice resolution in premature infants with jaundice. The procedure also provided an ideal therapeutic effect of diarrhea induced by phototherapy to promote compliance and maternal-infant bonding.

High glucose and high lipid induced mitochondrial dysfunction in JEG-3 cells through oxidative stress.

Few studies focused on the roles of high glucose combined with high lipid in placental development or fetal growth. This study was designed to investigate the roles of high glucose combined with high lipid in mitochondrial dysfunction of JEG-3 cells. We determined the cellular proliferation and apoptosis, superoxide dismutase (SOD) activity, concentration of malondialdehyde (MDA), and lactic acid dehydrogenase in control group, high glucose group, high lipid group, and high glucose and high lipid group, together with the mitochondrial dysfunction, Nrf2, HO-1, SMAC, and cytochrome C (Cyt-C) expression. Significant decrease of SOD and significant elevation of MDA was seen in high glucose and high lipid group compared with the other three groups. There was significant decrease in mitochondrial SMAC and Cyt-C in high glucose group, high lipid group, and high glucose and high lipid group compared with those of control group. Nrf2 and HO-1 protein expression in high glucose combined with high lipid group showed significant decrease compared with that of high lipid group or high glucose group. We speculated that combination of high glucose and high lipid induced oxidative stress in JEG-3 cells, and Nrf2/ARE pathway may be related to this process.

Impact of Saccharomyces boulardii on jaundice in premature infants undergoing phototherapy.

OBJECTIVES: To evaluate the therapeutic effect of Saccharomyces boulardii supplementation on jaundice in premature infants undergoing phototherapy. METHODS: In this article, the authors reviewed 100 hospitalized jaundiced premature infants under 35 weeks of gestational age. All infants were assigned to a control group (n = 45) and a treatment group (n = 55) randomly. The infants in the treatment group received S. boulardii supplementation by undergoing phototherapy and the infants in the control group were only treated by phototherapy. The total serum bilirubin levels were detected before and at the end of phototherapy, and transcutaneous bilirubin levels were measured on the 1st, 4th, 8th and 15th day of treatment. The duration of jaundice resolution and phototherapy, stool frequency, and characteristics were compared after phototherapy. RESULTS: The duration of jaundice resolution and phototherapy were shortened. Total serum bilirubin level was lower than the control group at the end of phototherapy (p < 0.05). Transcutaneous bilirubin levels decreased more significantly on the 8th and 15th day of treatment (p < 0.05), while there were no significant differences on the post-treatment 1st and 4th day (p > 0.05). In addition, bowel movements including stool frequency and Bristol Stool Form Scale ratings of stools also improved after treatment. CONCLUSIONS: S. boulardii in combination with phototherapy is effective and safe in reducing bilirubin levels and duration of phototherapy, accelerating jaundice resolution in premature infants with jaundice. The procedure also provided an ideal therapeutic effect of diarrhea induced by phototherapy to promote compliance and maternal-infant bonding.

Genetic Analysis of the LOXHD1 Gene in Chinese Patients With Non-Syndromic Hearing Loss.

Non-syndromic hearing loss (NSHL) is a common neurosensory disease with an extreme genetic heterogeneity which has been linked to variants in over 120 genes. The LOXHD1 gene (DFNB77), encoding lipoxygenase homology domain 1, is a rare hearing loss gene found in several populations. To evaluate the importance of LOXHD1 variants in Chinese patients with NSHL, we performed genetic analysis on LOXHD1 in 2,901 sporadic Chinese patients to identify the aspect and frequency of LOXHD1 causative variants. Next-generation sequencing using a custom gene panel of HL was conducted on 2,641 unrelated patients and whole-exome sequencing on the remaining 260 patients. A total of 33 likely causative variants were identified in 21 patients, including 20 novel variants and 13 previously reported pathogenic variants. Each of the 20 novel variants was evaluated according to ACMG criteria. These findings showed that causative variants in LOXHD1 were found in about 0.72% (21/2,901) of Chinese NSHL patients. This study is by far the largest number of novel variants identified in this gene expanding the range of pathogenic variants in LOXHD1, and suggests that variants in this gene occur relatively commonly in Chinese NSHL patients. This extensive investigation of LOXHD1 in Chinese NSHL patients proposed six recurrent LOXHD1 variants. These findings may assist in both molecular diagnosis and genetic counseling.

Hearing Phenotypes of Patients with Hearing Loss Homozygous for the GJB2 c.235delc Mutation.

Hereditary hearing loss is characterized by remarkable phenotypic heterogeneity. Patients with the same pathogenic mutations may exhibit various hearing loss phenotypes. In the Chinese population, the c.235delC mutation is the most common pathogenic mutation of GJB2 and is closely related to hereditary recessive hearing loss. Here, we investigated the hearing phenotypes of patients with hearing loss associated with the homozygous c.235delC mutation, paying special attention to asymmetric interaural hearing loss. A total of 244 patients with the GJB2 c.235delC homozygous mutation encountered from 2007 to 2015 were enrolled. The severity of hearing loss was scaled with the American Speech-Language-Hearing Association (ASHA). Auditory phenotypes were analyzed, and three types of interaural asymmetry were defined based on audiograms: Type A (asymmetry of hearing loss severity), Type B (asymmetry of audiogram shape), and Type C (Type A plus Type B). Of the 488 ears (244 cases) examined, 71.93% (351) presented with profound hearing loss, 14.34% (70) with severe hearing loss, and 9.43% (46) with moderate to severe hearing loss. The most common audiogram shapes were descending (31.15%) and flat (24.18%). A total of 156 (63.93%) of the 244 patients exhibited asymmetric interaural hearing loss in terms of severity and/or audiogram shape. Type A was evident in 14 of these cases, Type B in 106, and Type C in 36. In addition, 211 of 312 ears (67.63%) in the interaural hearing asymmetry group showed profound hearing loss, and 59 (18.91%) exhibited severe hearing loss, with the most common audiogram shapes being flat (27.88%) and descending (22.12%). By contrast, in the interaural hearing symmetry group, profound hearing loss was observed in 140 ears (79.55%), and the most common audiograms were descending (46.59%) and residual (21.59%). Hearing loss associated with the GJB2 c.235delC homozygous mutation shows diverse phenotypes, and a considerable proportion of patients show bilateral hearing loss asymmetry.

Prelingual Sensorineural Hearing Loss Caused by a Novel GJB2 Dominant Mutation in a Chinese Family.

BACKGROUND: GJB2 mutation is the most common cause of genetic deafness. Many pathogenic variations have already been identified, and thus, fewer and fewer novel pathogenic variations remain to be identified. Here, we describe a novel pathogenic variation associated with dominant hereditary deafness in a Chinese family. METHODS: In this study, we examined four generations of a Chinese family (M127) with hearing loss. Temporal CT scan, complete physical examination (including skin and hair), and audiological tests were performed. Targeted next-generation and Sanger sequencing were used to identify pathogenic mutations in affected individuals. RESULTS: All patients exhibited prelingual nonsyndromic sensorineural hearing loss, with severity ranging from moderate to severe. A novel dominant pathogenic variant c.205T > C (p.Phe69Leu) was identified in all patients in this family. CONCLUSIONS: c.205T > C (p.Phe69Leu) was identified as a novel dominant pathogenic variant of GJB2 associated with prelingual nonsyndromic sensorineural hearing loss.GJB2 mutation is the most common cause of genetic deafness. Many pathogenic variations have already been identified, and thus, fewer and fewer novel pathogenic variations remain to be identified. Here, we describe a novel pathogenic variation associated with dominant hereditary deafness in a Chinese family.

Prepregnancy maternal diabetes combined with obesity impairs placental mitochondrial function involving Nrf2/ARE pathway and detrimentally alters metabolism of offspring.

Metabolic disorders usually increase the level of reactive oxygen species (ROS) and damage mitochondrial function. The placenta supplies nutrients and hormonal signals to the fetus for regulating fetal metabolism, and is also prone to injury by oxidants. The aim of this study was to determine the effect of pre-existing maternal type 2 diabetes mellitus (DM) combined with obesity on placental mitochondrial function and metabolism disorders of offspring. The study included 96 pregnant women. The women were put into the following groups: healthy women (control, n=24), women with DM (DM, n=24), women with obesity (OB, n=24) and women with both DM and obesity (DM+OB, n=24). The ROS level, mitochondrial content, and the mitochondrial respiratory complex activities of the placenta were measured in the four groups. The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) was detected by immunofluorescence staining and western blotting. In addition, serum levels of insulin, glucose, leptin, nonesterified fatty acid (NEFA), adiponectin and triglycerides of their offspring were also measured. Maternal DM combined with obesity markedly increased ROS level, reduced mitochondrial DNA (mtDNA) content and mitochondrial respiratory complex I, II-III activities in placenta compared to the placenta from the control group and the DM group. Maternal DM combined with obesity significantly decreased Nrf2 and HO-1 expression. Furthermore, maternal DM combined with obesity influenced the glucose and lipid metabolism in their offspring. In conclusion, women with both DM and obesity detrimentally alter placenta function in oxidative stress regulation, and the Nrf2/ARE (antioxidant responsive element) pathway is involved. This may increase metabolic disturbance susceptibility in their offspring.

Prognosis of psychomotor and mental development in premature infants by early cranial ultrasound.

BACKGROUND: It is of high incidence of brain injuries in premature infants, so it is necessary to diagnose and treat the brain injury early for neonatal clinical practice. We are aimed to investigate the relationship between early postnatal cranial ultrasonography and psychomotor and mental development in prematrue infants at the age of 12 months. METHODS: Two-hundred and eight premature infants were selected and underwent follow-up from January, 2007 to November, 2012. Cranial ultrasonography was performed on them. The developmental outcomes of these premature infants at the age of 12 months were assessed by the psychomotor developmental index (PDI) scale and mental development index (MDI). The relationship between ultrasonic gray-scale value and PDI and MDI was analyzed. RESULTS: The worse prognosis for psychomotor and mental development was associated with the gestational age, Apgar score(1 min), gender, chorioamnionitis, duration of mechanical ventilation and duration of mechanic ventilation. The differences between the prognosis of psychomotor and mental development, and peri-intraventricular hemorrhage (PIVH) and periventricular white matter damage (PWMD), were statistically significant (P<0.05). There were also significant differences between the early postnatal ultrasonic gray-scale value and prognoses of both psychomotor development and mental development (P<0.05). There were negative correlations between ultrasonic gray-scale and both PDI and MDI (r=-0.753, P<0.05; r=-0.764, P<0.05). CONCLUSIONS: The early postnatal cranial ultrasonography can assist to predict the prognosis of psychomotor and mental development for premature infants. The higher grade of PIVH and PWMD was associated with the worse prognosis of psychomotor and mental development.

Dynamic changes of cerebral-specific proteins in full-term newborns with hypoxic-ischemic encephalopathy.

The aim of this study was to observe the dynamic changes of serum brain-derived neurotrophic factor (BDNF), S-100B, and Tau proteins levels in full-term newborns with hypoxic-ischemic encephalopathy (HIE) and to discuss their significance in brain damage. Serum samples of 28 full-term newborns diagnosed with HIE and 20 controls were obtained in the first 24 h of life. Another serum samples were also taken, respectively, at 3 and 7 days of life in HIE group. The concentrations of BDNF, S-100B, and Tau proteins were measured by the enzyme-linked immunosorbent assay method. Mean concentrations of BDNF, S-100B, and Tau proteins among different time period and in different grades of HIE group were calculated and compared. Compared with the control group, serum BDNF and proteins S-100B levels in HIE group were significantly elevated in 24 h after birth (P < 0.05) and their concentrations were also significantly higher among patients with mod-severe HIE compared to those with mild HIE at 24 h and 7 days after asphyxia (P < 0.05). Regardless of whether mod-severe HIE or mild HIE, there were no significant difference of serum BDNF and proteins S-100B among the three different time periods. There was no difference in Tau protein levels between HIE group and control group, also no difference between mod-severe HIE group and mild HIE group. BDNF and proteins S-100B are up-regulated early in asphyxia neonates with HIE; and the released amount of BDNF and proteins S-100B from nerve center system correlate with the extent of encephalopathy.