Objective To explore the efficacy and safety of bedside continuous blood purification (CBP) in the treatment of neonatal multiple organ failure (MOF).Methods Totally 6 newborn infants of MOF were hospitalized in department of neonatology in our hospital from June 2011 to June 2013.These 6 cases of clinical data were retrospectively analyzed,6 neonates were treated with CBP combined with conventional treatment.The model for CBP was continuous veno-venous hemodialysis filtration (CVVHDF),blood flow velocity was 3 to 5 ml/(kg· min),replacement fluid dose was 20 to 30 ml/(kg· h),dialysis fluid dose was 15 to 25 rnl/(min· m2).The clinical outcome measures included,blood pressure,blood pH,K+,Na+,blood urea nitrogen,creatinine,urine volume,PaO2/FiO2 and epinephrine intravenous dose,respectively before CBP treatment,6 h,12 h,24 h,48 h after CBP treatment and the end of CBP treatment.The efficacy of CBP treatment was evaluated in neonatal MOF.Results Gestational age of 6 neonates with MOF was 33 to 41 weeks,2 to 19 days old,2.25 to 3.36 kg birth weight.Primary disease was 4 cases of neonatal septicemia(1 case with congenital hereditary metabolic disease),2 cases of severe neonatal asphyxia.All 6 cases of venous catheter were smoothly done.CBP treatment persisted for 49 to 106 hours.Compared with before CVVHDF treatment,blood K+,blood urea nitrogen,creatinine significantly decreased at 12 h after CVVHDF treatment [(5.32 ± 1.84) mmol/L vs.(9.81 ±3.61) mmol/L,(9.0 ±3.4) mmol/L vs.(12.8 ±6.1) mmol/L,(99 ± 16) μmol/L vs.(176 ±25) μmol/L,P ＜0.05],and reached the normal range at 24 h after treatment,urine volume significantly increased at 24 h after treatment (P ＜ 0.05).PaO2/FiO2 reached 200 mmHg (1 mmHg =0.133 kPa) at 6 h after treatment and more than 300 mmHg at 24 h after treatment(P ＜0.05).Fifty percent of epinephrine intravenous dose were down-regulation at 12 h after treatment and stopped using epinephrine at 48 h after treatment.CBP treatment of 6 cases showed effective.Conclusion Application of bedside CBP treatment in neonatal MOF is safe,can effectively help neonates with MOF to skip over renal failure stage.

Between May 2009 and February 2012,60 patients scheduled for laparoscopic appendectomy were randomly assigned to receive either transumbilical single-port laparoscopic appendectomy (TUSPLA) or three-port laparoscopic appendectomy (TPLA).The operative duration was significantly longer in TUSPLA than TPLA cases.No statistically significant differences existed in hospital stay,visual analog scale (VAS) score,analgesia requirement or complication rate.The abdominal scar was inconspicuous for TUSPLA.It suggested that TUSPLA with conventional instrumentation is technically feasible and safe with scarless healing.

Objective To explore the efficacy and safety of mild hypothermia (MH) in treating the infants with moderate-to-severe neonatal hypoxic-ischemic encephalopathy(HIE),and to make a follow-up of the nerve motor development of the infants at 18 months old after discharge.Methods Totally 61 neonates with moderate-to-severe HIE in Neonatal Intensive Care Unit (NICU) from Jan.2007 to Dec.2013 were retrospectively analyzed.According to before and after MH therapeutic apparatus was used by NICU of Shanghai Children's Hospital,61 neonates of HIE were divided into 2 groups,the conventional treatment group(25 cases) and MH treatment group(36 cases).The patients in both groups were measured respectively by using the amplitude integrated electroencephalography (aEEG) before MH treatment and at 72 hours after M H treatment,by neonatal behavioral neurological assessment(NBNA) on the 28th day after birth,and by adopting Bayley Scales of Infant Development at 18 months old.The adverse reactions,serious disability cases and deaths of MH treatment were recorded.Results Compared with the conventional treatment group,aEEG recording before treatment showed no statistically significant differences in MH treatment group [maximum voltage:(22.4 ±3.1) μV vs(18.6 ±2.5) μV,maximum voltage:(8.2 ±2.6)μV vs(6.5 ±1.9) μV,t =1.264,0.852,all P ＞ 0.05].However,aEEG recording at 72 h after treatment showed statistically significant differences in MH treatment group [maximum voltage:(24.1 ± 3.2) μV vs (30.6 ± 2.8) μV,maximum voltage:(9.7 ± 3.4) μV vs (13.3 ± 2.2) μV,t =6.376,4.257,all P ＜ 0.05].Severe disability cases [24.0％ (6/25 cases) vs 5.6％ (2/36 cases),x2 =4.405,P ＜ 0.05] and deaths [16.0％ (4/25 cases) vs 0 (0/36 case),x2 =6.1 64,P ＜ 0.05] in MH treatment group were significantly decreased,and there was significantly difference in NBNA on the 28th day after birth[(35.9 ± 2.1) vs(39.1-± 1.6),t =3.361,P ＜ 0.05],and scales of neurobehavioral evaluation through follow-up of 18 months old [mental development index (MDI):(85.2 ± 10.7) vs (96.5-± 13.1),t =7.839,P ＜ 0.05].Very few neonates had apnea,coagulation dysfunction,arrhythmia and other adverse reactions in MH treatment course.Conclusions MH treating moderate-to-severe HIE is safe and effective.MH is effective in reducing death and major disabilities in neonates with moderate-to-severe HIE and without significant side effects.MH can obviously improve the development of nervous system disorders in 0-18 months infants,and can significantly improve these infants' Bayley developmental scale neurobehavioral scores.

Objective:To investigate the clinical and molecular genetic characteristics of 6q24-related transient neonatal diabetes mellitus (6q24-TNDM).Methods:The clinical data of two neonates with 6q24-TNDM admitted to Shanghai Children's Hospital, Shanghai Jiao Tong University in 2017, were retrospectively collected. The methylation levels of 16 cytidine-phosphate-guanosine (CpG) sites from the methylated differentially modified region (DMR) in 6q24 were quantitatively analyzed by pyrosequencing.Results:Case 1, aged 5 d, was born at 37 +4 gestational weeks due to fetal growth restriction, and case 2 was 11-days old and born at 38 +2 gestational weeks. Both infants were male and small for age. They were born through a cesarean section. The birth weight of case 1 and case 2 were 2 340 g and 2 600 g, respectively. They were admitted due to hyperglycemia with blood glucose of 12.95 and 8.00 mmol/L on admission, respectively. Physical examination showed slightly poor skin elasticity and thin subcutaneous fat. Laboratory examination revealed lower serum insulin (<1.39 and 3.94 pmol/L) and peptide C (0.05 and 0.14 nmol/L) levels, positive results of urine glucose, negative tests for urine ketone, serum anti-glutamic acid decarboxylase antibody, anti-insulin antibody, and islet cell antibody in both cases. Normal size of the pancreas was observed by ultrasonography. The infants were improved and were discharged after subcutaneous insulin infusion for more than two weeks. The treatment was discontinued at 69 d and 42 d postnatally for case 1 and case 2. Prenatal diagnosis of the two infants showed normal karyotypes and uniparental disomy of chromosome 6 indicated by single nucleotide polymorphism chip. No pathogenic mutations were detected by next-generation sequencing after admission. The methylation levels of 16 CpG sites in DMR of 6q24 in the two cases, which were quantitatively analyzed by pyrosequencing, were lower than 10% (normal value in healthy matched controls: 40%), indicating an obvious hypomethylation. Conclusions:For children with TNDM who are small for gestational age at birth, presenting hyperglycemia with decreased serum insulin and C-peptide levels, pyrosequencing can be used to quantitatively analyze the methylation levels of CpG sites in 6q24 DMR, which can quickly and directly assist in the diagnosis of 6q24-TNDM, thereby contributing to the treatment and prognosis assessment.

Objective: To explore the abnormal changes in neuroelectric activity in the primary visual cortex of rats deprived of vision in one eye and to investigate the regulatory effect of acupuncture in the sensitive period on the abnormal coding and conduction of electrical signals of rats' optic neurons.Methods: Sixty 14-day-old Sprague-Dawley rats were randomly divided into a blank group, a model group, an early-stage acupuncture group, a middle-stage acupuncture group, and a late-stage acupuncture group, with 12 rats in each group. Rats in every group except the blank group received right eyelid suturing to create a monocular deprivation model in the sensitive period of visual development (from the day rats open their eyes to the 45th day after their birth). Rats in the three acupuncture groups started to undergo acupuncture respectively on the 3rd, 12th, and 21st days after the model replication was done, with each group receiving nine-day treatment. The activity level of the neuroelectrical signal of the primary visual cortex in each group, including the latency and amplitude of P100 wave, average discharge frequency and amplitude of neurons, the power spectral density (PSD), and interspike interval (ISI), were measured by neuroelectric evaluation technology after the acupuncture treatment was finished. Results: Compared with the blank group, the latency of P100 wave in the visual center of vision-deprived eyes was significantly prolonged, and the amplitude was significantly reduced (P<0.05); the average discharge frequency and amplitude of the neurons in the visual cortex also decreased significantly (P<0.05); PSD decreased and ISI was prolonged significantly (P<0.05). Compared with the model group, the abnormal electrical activity of optic neurons in the three acupuncture groups ameliorated, the latency of P100 shortened, and the amplitude of P100 increased (P<0.05), the discharge frequency and amplitude increased significantly (P<0.05), the PSD reduced, and the ISI shortened (P<0.05). In addition, among the three acupuncture groups, the early-stage acupuncture group had the best effect on various indicators. Conclusion: Abnormal electrophysiological activity is significant in the visual center of vision-deprived rats, and acupuncture treatment in the sensitive period of visual development can enhance the bioelectrical activity of visual nerve cells, improve the efficiency of optic nerve conduction, and regulate the inhibition and retardation of visual response caused by visual deprivation.

OBJECTIVETo carry out rapid genetic diagnosis for a child affected with citrullinemia type Ⅰ.

METHODSPeripheral venous blood samples were obtained from the two-day-old child and his parents as well as 100 healthy controls. Serum ammonia and citrulline was determined by biochemical test and tandem mass spectrometry. Sixteen pairs of primers were designed for high-resolution melting (HRM) analysis of all exons and adjacent intronic sequences of the ASS1 gene in the proband, parents and healthy controls. Suspected mutations were confirmed by DNA sequencing, while the mRNA transcripts of the ASS1 gene were determined by reverse transcription (RT)-PCR. Functional impact of the mutation sites was predicted with PolyPhen-2 and SIFT Blink software.

RESULTSBlood ammonia and citrulline of the proband have respectively reached 286 μmol/L and 487.69 μmol/L, which far superseded the normal values. HRM analysis and DNA sequencing have identified in the child a homozygous c.380G>A (p.R127Q) mutation in exon 6 of the ASS1 gene, in addition with a homozygous IVS8+60G>A substitution in intron 8, while his parents were heterozygous carriers for both mutations. RT-PCR assay indicated that the IVS8+60G>A mutation did not result in abnormal splicing of the ASS1 gene transcripts. Bioinformatic analysis suggested that the site for p.R127Q was conserved among 45 species of vertebrates and may play a crucial role in citrulline metabolism.

CONCLUSIONThe severe urea cycle disorder in the proband was probably due to the compound homozygous R127Q and IVS8+60G>A mutations of the ASS1 gene.

Objective:To study the early predictive value of urine neutrophil gelatinase-associated lipoprotein (NGAL) and kidney injury molecule-1 (KIM-1) for acute kidney injury (AKI) in neonates with severe asphyxia.Method:From January 2019 to June 2020, neonates with severe asphyxia admitted to our hospital within 6 hours after birth were enrolled in the study. The dynamic changes of urine NGAL and KIM-1 at admission, 24 h, 48 h and 1 w after birth were examined. Neonates were assigned into AKI group and non-AKI group according to the clinical practice guidelines for AKI issued by KDIGO (Kidney Disease: Improving Global Outcome). The sensitivity and specificity of NGAL and KIM-1 predicting AKI at different time points were evaluated using ROC curve and area under curve (AUC).Result:According to the diagnostic criteria of neonatal AKI, 9 cases were in the AKI group and 42 cases in the non-AKI group, and the incidence of AKI was 17.6%. Urine NGAL was significantly increased in AKI group at admission and 24 h after birth compared with the non-AKI group [(115.6±75.5) ng/ml vs. (49.8±29.0) ng/ml, (90.7±35.6) ng/ml vs. (55.6±30.7) ng/ml] ( P<0.05). No significant differences existed at 48 h and 1 w after birth between the two groups. At 24 h after birth, urine KIM-1 in the AKI group was significantly higher than the non-AKI group [(808.3±555.3) pg/ml vs. (318.4±234.0) pg/ml, P<0.05] and no significant differences existed between the two groups at admission, 48 h and 1 w after birth. The AUC of NGAL predicting AKI at admission, 24 h, 48 h and 1w after birth were 0.804 (95% CI 0.573~1.000), 0.792 (95% CI 0.580~1.000), 0.732 (95% CI 0.517~0.947) and 0.551(95% CI 0.371~0.730), respectively. The AUC of KIM-1 predicted AKI at admission, 24 h, 48 h and 1 w after birth was 0.860 (95% CI 0.676~1.000), 0.824 (95% CI 0.655~0.993), 0.768 (95% CI 0.622~0.914), 0.622 (95% CI 0.392~0.852), respectively. Conclusion:At admission, 24 h and 48 h after birth, urine NGAL and KIM-1, as kidney injury markers, may predict the occurrence of AKI after severe neonatal asphyxia.