Journal of Pediatric Diseases

Abstract To observe the expression of neuropilin-1 (NRP-1) induced by paired-box gene 2 (PAX2) during the process of epithelial-to-mesenchymal transition (EMT) and renal fibrosis in unilateral ureteral obstruction (UUO) model in rats, and explore the mechanism of EMT induced by PAX2. Methods : The recombinant lentivirus expression vector for PAX2 was constructed and transfected into rat normal renal tubular epithelial cell line (NRK52E). The experimental cells were divided into three groups: transfection group, empty vector group, and normal group. E-cadherin and α-SMA were detected by western blot and real-time PCR. Expression of NRP-1 was detected by western blot, real-time PCR, and immunofluorescence. Sixty male Wistar rats were randomly divided into two groups: the sham-operation group (n=30) underwent left ureteral dissection, the UUO group (n=30) underwent left ureteral ligation. Post-operation on days 3, 7, 14, 21 and 28, 6 rats from each of the groups were sacrificed and the obstructed kidneys were dissected out. The histopathological changes were observed by hematoxylin-eosin and Masson staining. E-cadherin and α-SMA were detected by western blot and immunohistochemistry. Expression of NRP-1 and PAX2 were determined by western blot, immunohistochemistry, and real-time PCR. Results : Expression of NRP-1 mRNA and protein and α-SMA protein increased (P<0.05) while E-cadherin protein expression decreased (P<0.05) in the transfection group as compared to the empty vector group in vitro. In the UUO group, fibrosis was obvious, and there was decreased expression of E-cadherin protein (P<0.05) and increased expression of α-SMA protein and NRP-1 mRNA and protein (P<0.05) in comparison to the sham group. Conclusion : NRP-1 maybe mediate PAX2-induced EMT in renal tubular epithelial cells and renal fibrosis.

Abstract Objective Autosomal recessive polycystic kidney disease (ARPKD) is a rare inherited renal cystic disease involving multiple organs. It is caused by mutations in the PKHD1 gene. Here, we investigate the gene mutations in a family affected by ARPKD. Methods Genomic DNA was extracted from peripheral blood leukocytes obtained from the subjects, by means of targeted gene capture and next generation sequencing technologies for mutation screening, and were confirmed by Sanger sequencing. Results Two heterozygous mutations of PKHD1 , c.6890T>C (p.Ile2297Thr) and c.11215C>T (p.Arg3739Trp), located in exons 43 and 62, respectively, were identified in the patient. Furthermore, the father and mother were revealed to be carriers of heterozygous c.6890T>C (p.Ile2297Thr) and c.11215C>T (p.Arg3739Trp) mutations, respectively. Mutation of c.11215C>T (p.Arg3739Trp) has been found in the ARPKD Mutation Database ( http://www.humgen.rwth-aachen.de ) but mutation of c.6890T>C (p.Ile2297Thr) has not been reported. Conclusions Compound heterozygous PKHD1 mutations were elucidated to be the molecular basis of ARPKD in this patient. The newly identified c.6890T>C (p.Ile2297Thr) mutation in the patient expands the mutation spectrum of the PKHD1 gene. Targeted gene capture and next generation sequencing are suitable for genetic diagnosis of single-gene inherited diseases like ARPKD, in which the pathogenic gene is large.

Abstract Objective : To improve the efficiency of lumbar intervertebral discs protrusion by determining the ultrasound biomarkers. Materials and Methods : The study included 68 patients with the disc protrusion and 65 healthy adolescents with normal lumbar intervertebral discs and neurologic status aged 16-18 years. Ultrasonography (USG) was performed at the level of disks L1-L2, L2-L3, L3-L4, L4-L5, L5-S1 in longitudinal and transverse projections. In longitudinal section was measured height of lumbar vertebrae and intervertebral discs, in axial section – the sagittal sizes of intervertebral discs and spinal canal, width of spinal nerve canals, thickness of the yellow ligament. Results : In 31 (45,6±6,0%) cases the protrusion was paramedian, in 16 (23,5±5,1%) – posterolateral, in 13 (19,1±4,8) – median and in 8 (11,8±3,9%) – circular types. The paramedian protrusion was significantly more frequently recorded than the posterolateral (P<0,05), median and circular (p <0.001) types. In 7 (10,3±3,7% ) cases the protrusion was localized at the level of L2-L3, in 13 (19,1 ±4,8%) – L3-L4, in 27 (39,7±5,9%) - at the level of L4-L5 and in 21 (30,9±5,6%) – at the level of L5-S1 respectively. There were no significant differences in the frequency of occurrence between L2-L3 and L3-L4, as well as L4-L5 and L5-S1. The lower located lumbar discs were significantly more affected than the upper ones (P<0,05; P<0,001). Conclusions: In adolescents in the lumbar spine, paramedian protrusion are most commonly found, which are most often localized at the level of both L4-L5 and L5-S1. The greatest  narrowing and deformation of the spinal nerve canal is observed by posterolateral and paramedian protrusion. The greatest thickness of the yellow ligament, radiculopathy  is observed at level of L5-S1 protrusion. 

Abstract Introduction:  dietary therapy plays an important role in keeping good blood glucose level in patients with type 1 diabetes (T1D).  We evaluated the influence of carbohydrate counting  (CC) on auxological parameters and metabolic management in children and adolescents with T1D. Methods and Materials: we enrolled   all pediatric patients with T1DM followed up in our outpatient Pediatrics Diabetology Unit at IRCCS Policlinic San Matteo of Pavia, between June 2016 and June 2017. In all subjects the diagnosis was made or confirmed at least 6 months before this study started. All subjects were treated with a basal bolus insulin regimen by means of multiple daily injections or continuous subcutaneous insulin infusion with pump. Subjects were classified as “adherent” or “not adherent” to CC; auxological (weight, height, BMI score and BMI-z score) and metabolic parameters (glycated hemoglobin (HbA1c), insulin needs/24h, insulin to carbohydrates ratio (ICR), insulin-sensitivity factor (ISF)) were recorded Results:  Of all the patients enrolled (n=134; 75M/ 59F; mean age 12.6±5.1 years) diagnosed with T1DM, 21 (15.7%) were adherent to CC (16 of them were on pump insulin therapy, 5 on multiple injection insulin therapy). HbA1c was not statistically different in the two groups (p=0.46 and p=0.6, respectively). Patients adherent to CC tended to show higher BMIs (p=0.005) as well as BMI z-score (p=0.04) and overweight (p=0.03) than the non adherent group. the non adherent group showed lower ICR (p=0.04) and ISF (p=0.04), significantly related to the BMI (p<0.001). Conclusion:  CC represents a good alternative to the classic nutritional treatment in children and adolescents diagnosed with T1DM. Moreover, CC may influence weight and metabolic management, so it is necessary to be appropriately trained on the adequate dietary recommendations and the right counting method. 

Abstract A male newborn admitted in the Neonatal Intensive Care Unit due to dyspnea and cyanosis. The baby was intubated due to tachypnea. No murmurs were heard on auscultation.The ultrasound of the fetal heart before birth showed cardiac malformations. Chest X-ray showed : Increased pulmonary vascular markings and cardiomegaly. The abdominal X-ray showed normal liver, spleen and intestine. Electrocardiogram showed Sinus rhythm and tachycardia. On the first day after birth, two-dimensional echocardiography demonstrated marked hypertrophy of both ventricles (the posterior wall of the left ventricle was 33mm thick). The baby was started on treatment with low flow oxygen support, digoxin and captoril to enhance myocardial contractility, creatine phosphate for myocardial nutrition , furosemide diuretic to reduced load, enhance feeding, monitor bilirubin, prevent neonatal jaundice, and close attention was paid to the disease changes. The baby was stable and was discharged from the hospital. After 20days of discharge, the baby was admitted again with complains of shortness of breath and cyanosis after 20 days of discharge. The heart beat was low on auscultation with alternating tachycardia and bradycardia, with an occasional gallop rhythm. The baby was kept on ventilator assisted ventilation with the required parameters and necessary investigations were performed. On repeating the two-dimensional echocardiography, the left ventricular posterior wall and the ventricular septum was increased compared to the previous echocardiography. A mutation on Chromosome 18, c.1921G>A was detected on gene mutational analysis. Recently, some genetic studies have shown that mutations in chromosomes 1, 11, 14 and 15, and mutations in sarcomere proteins genes are autosomal dominant