V-A ECMO for neonatal coxsackievirus B fulminant myocarditis: a case report and literature review.

Background: Neonatal (enteroviral) myocarditis (NM/NEM) is rare but unpredictable and devastating, with high mortality and morbidity. We report a case of neonatal coxsackievirus B (CVB) fulminant myocarditis successfully treated with veno-arterial extracorporeal membrane oxygenation (V-A ECMO). Case presentation: A previously healthy 7-day-old boy presented with fever for 4 days. Progressive cardiac dysfunction (weak heart sounds, hepatomegaly, pulmonary edema, ascites, and oliguria), decreased left ventricular ejection fraction (LVEF) and fractional shortening (FS), transient ventricular fibrillation, dramatically elevated creatine kinase-MB (405.8 U/L), cardiac troponin I (25.85 ng/ml), and N-terminal pro-brain natriuretic peptide (NT-proBNP > 35,000 ng/L), and positive blood CVB ribonucleic acid indicated neonatal CVB fulminating myocarditis. It was refractory to mechanical ventilation, fluid resuscitation, inotropes, corticosteroids, intravenous immunoglobulin, and diuretics during the first 4 days of hospitalization (DOH 1-4). The deterioration was suppressed by V-A ECMO in the next 5 days (DOH 5-9), despite the occurrence of bilateral grade III intraventricular hemorrhage on DOH 7. Within the first 4 days after ECMO decannulation (DOH 10-13), he continued to improve with withdrawal of mechanical ventilation, LVEF > 60%, and FS > 30%. In the subsequent 4 days (DOH 14-17), his LVEF and FS decreased to 52% and 25%, and further dropped to 37%-38% and 17% over the next 2 days (DOH 18-19), respectively. There was no other deterioration except for cardiomegaly and paroxysmal tachypnea. Through strengthening fluid restriction and diuresis, and improving cardiopulmonary function, he restabilized. Finally, notwithstanding NT-proBNP elevation (>35,000 ng/L), cardiomegaly, and low LVEF (40%-44%) and FS (18%-21%) levels, he was discharged on DOH 26 with oral medications discontinued within 3 weeks postdischarge. In nearly three years of follow-up, he was uneventful, with interventricular septum hyperechogenic foci and mild mitral/tricuspid regurgitation. Conclusions: Dynamic cardiac function monitoring via real-time echocardiography is useful for the diagnosis and treatment of NM/NEM. As a lifesaving therapy, ECMO may improve the survival rate of patients with NM/NEM. However, the "honeymoon period" after ECMO may cause the illusion of recovery. Regardless of whether the survivors of NM/NEM have undergone ECMO, close long-term follow-up is paramount to the prompt identification and intervention of abnormalities.

A meta-analysis of randomized controlled trials: combination of ketamine and propofol versus ketamine alone for procedural sedation and analgesia in children.

Although ketamine has been used for procedural sedation and analgesia, some researchers have assessed ketamine-propofol as a better alternative because of its reduced adverse events. The goal of this review was to compare adverse events between ketamine-propofol and ketamine for procedural sedation and analgesia in children. We searched the literature from their inception to May 2018 without the restriction of language. We included all randomized controlled trials comparing ketamine-propofol with ketamine for procedural sedation and analgesia in children. The meta-analysis was conducted using the Stata software. A total of six studies involving 693 individuals were included. Pooling of data showed that subjects with ketamine-propofol had similar incidence of respiratory adverse events compared to those with ketamine (RR 1.16, 95% CI 0.68-1.98). However, ketamine-propofol was effective in reducing cardiovascular adverse events compared to ketamine (RR 0.11, 95% CI 0.04-0.31). Ketamine-propofol was also effective in reducing psychomimetic adverse events compared to ketamine (RR 0.39, 95% CI 0.16-0.93). In regard to nausea and vomiting, ketamine-propofol was significantly effective (RR 0.43, 95% CI 0.25-0.74). In addition, we could not demonstrate differences in efficacious sedation between ketamine-propofol and ketamine. Although our study was not able to demonstrate differences in efficacious sedation between ketamine-propofol and ketamine, we confirmed that ketamine-propofol sedation had a lower frequency of adverse events compared to ketamine sedation in children.

Efficient Mining of Variants From Trios for Ventricular Septal Defect Association Study.

Ventricular septal defect (VSD) is a fatal congenital heart disease showing severe consequence in affected infants. Early diagnosis plays an important role, particularly through genetic variants. Existing panel-based approaches of variants mining suffer from shortage of large panels, costly sequencing, and missing rare variants. Although a trio-based method alleviates these limitations to some extent, it is agnostic to novel mutations and computational intensive. Considering these limitations, we are studying a novel variants mining algorithm from trio-based sequencing data and apply it on a VSD trio to identify associated mutations. Our approach starts with irrelevant k-mer filtering from sequences of a trio via a newly conceived coupled Bloom Filter, then corrects sequencing errors by using a statistical approach and extends kept k-mers into long sequences. These extended sequences are used as input for variants needed. Later, the obtained variants are comprehensively analyzed against existing databases to mine VSD-related mutations. Experiments show that our trio-based algorithm narrows down candidate coding genes and lncRNAs by about 10- and 5-folds comparing with single sequence-based approaches, respectively. Meanwhile, our algorithm is 10 times faster and 2 magnitudes memory-frugal compared with existing state-of-the-art approach. By applying our approach to a VSD trio, we fish out an unreported gene-CD80, a combination of two genes-MYBPC3 and TRDN and a lncRNA-NONHSAT096266.2, which are highly likely to be VSD-related.

[Effect of nitric oxide on the preterm neonatal rabbit ductus arteriosus cystathionine-γ-lyase/hydrogen sulfide pathway].

OBJECTIVE: To investigate the impact of sodium nitroprusside (a nitric oxide donor) in the ductus arteriosus in preterm rabbits on hydrogen sulfide (H(2)S)-cystathionine-γ-lyase (CSE) system. METHODS: For 16 Japanese white rabbits pregnant for 21 days were randomly divided into four groups, each of the following groups had 4 rabbits: control group, intraperitoneal injection of sodium nitroprusside 1 mg/kg, 2.5 mg/kg, and 5.0 mg/kg groups. The rabbits in control group had a peritoneal puncture with a simple hollow needle, and those in the other groups were given corresponding dose of intraperitoneal injection of sodium nitroprusside at gestational age 23 and 25 days, respectively. At gestational age 26 days the fetuses of the pregnant rabbits were removed surgically, and 28 fetal rabbits were obtained from the control group, 27 from the sodium nitroprusside small dose group, 29 from the medium dose group, and 26 from the large dose group. The fetal heart blood sample of 1 ml was taken from each fetus, and immediately after sampling the arterial ductal tissues were dissected. Fetal rabbit plasma proteins hydrogen sulfide content was determined by using de-protein method, and real time quantitative RT-PCR was used for determination of arterial tissue CSE gene and western-blotting was used for measuring protein expression of CSE. RESULTS: In control group hydrogen sulfide content of fetal rabbits plasma (55.68 ± 6.57) µmol/L and arterial tissue CSE mRNA expression was 1.07 ± 4.12; the parameters in intraperitoneal injection of sodium nitroprusside group 1 mg/kg were (60.02 ± 6.09) µmol/L and 3.46 ± 0.18; in intraperitoneal injection of sodium nitroprusside group 2.5 mg/kg, were (64.71 ± 7.12) µmol/L and 10.95 ± 0.22; and in intraperitoneal injection of sodium nitroprusside group 1 mg/kg were (70.63 ± 8.07) µmol/L and 19.56 ± 0.17. Comparison between small dose group and control group, medium dose group and small dose group, high dose group and medium dose group showed that the above data were significantly different P < 0.05, with the injection of sodium nitroprusside CSE protein expression increased gradually with increasing doses. CONCLUSION: Sodium nitroprusside showed an enhancing effect on preterm CSE-H(2)S system in rabbit ductus arteriosus in a certain range of concentration in a dose-dependent manner.

Homozygous disruption of the Tip60 gene causes early embryonic lethality.

Tat-interactive protein 60 (Tip60) is a member of the MYST family, proteins of which are related by an atypical histone acetyltransferase (HAT) domain. Although Tip60 has been implicated in cellular activities including DNA repair, apoptosis, and transcriptional regulation, its function during embryonic development is unknown. We ablated the Tip60 gene (Htatip) from the mouse by replacing exons 1-9 with a neomycin resistance cassette. Development and reproduction of wild-type and heterozygous animals were normal. However, homozygous ablation of the Tip60 gene caused embryolethality near the blastocyst stage of development, as evidenced by inability of cells in Tip60-null blastocysts to hatch and survive in culture. Monitoring cell proliferation and death by detecting EdU-substituted DNA and TUNEL labeling revealed suppression of cell proliferation concomitant with increased cell death as Tip60-null cells attempted to hatch from blastocysts. These findings indicate that Tip60 is essential for cellular survival during the blastocyst-gastrula transition of embryogenesis.

Mouse epidermal neural crest stem cell (EPI-NCSC) cultures.

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the physiological property to generate a wide array of differentiated cell types, including neurons, nerve supporting cells, smooth muscle cells, bone/cartilage cells and melanocytes. EPI-NCSC are easily accessible in the hairy skin and can be isolated as a highly pure population of stem cells. This video provides a detailed protocol for preparing mouse EPI-NCSC cultures from whisker follicles. The whisker pad of an adult mouse is removed, and whisker follicles dissected. The follicles are then cut longitudinally and subsequently transversely above and below the bulge region. The bulge is removed from the collagen capsule and placed in a culture plate. EPI-NCSC start to emigrate from the bulge explants 3 to 4 days later.

Epidermal neural crest stem cells (EPI-NCSC) and pluripotency.

This article serves three purposes. We summarize current knowledge of the origin and characteristics of EPI-NCSC, review their application in a mouse model of spinal cord injury, and we present new data that highlight aspects of pluripotency of EPI-NCSC. EPI-NCSC are multipotent stem cells, which are derived from the embryonic neural crest and are located in the bulge of hair follicles. EPI-NCSC can undergo self-renewal and they are able to generate all major neural crest derivatives, including neurons, nerve supporting cells, smooth muscle cells, bone/cartilage cells and melanocytes. Despite their ectodermal origin, neural crest cells can also generate cell types that typically are derived from mesoderm. We were therefore interested in exploring aspects of EPI-NCSC pluripotency. We here show that EPI-NCSC can fuse with adult skeletal muscle fibers and that incorporated EPI-NCSC nuclei are functional. Furthermore, we show that adult skeletal muscle represents an environment conducive to long-term survival of neurogenic EPI-NCSC. Genes used to create induced pluripotent stem (iPS) cells are present in our EPI-NCSC longSAGE gene expression library. Here we have corroborated this notion by real-time PCR. Our results show similarities in the expression of Myc, Klf4, Sox2 and Lin28 genes between EPI-NCSC and embryonic stem cells (ESC). In contrast there were major differences in Nanog and Pou5f1 (Oct-4) expression levels between EPI-NCSC and ESC, possibly explaining why EPI-NCSC are not tumorigenic. Overall, as embryonic remnants in an adult location EPI-NCSC show several attractive characteristics for future cell replacement therapy and/or biomedical engineering: Due to their ability to migrate, EPI-NCSC can be isolated as a highly pure population of multipotent stem cells by minimally-invasive procedures. The cells can be expanded in vitro into millions of stem cells/progenitors and they share some characteristics with pluripotent stem cells without being tumorigenic. Since the patients' own EPI-NCSC could be used for autologous transplantation, this would avoid graft rejection.

Microsurgical treatment of Nelson's syndrome.

OBJECTIVE: To discuss the etiology, diagnostic criteria and treatment of Nelson's syndrome. METHODS: Twenty-three patients with Nelson's syndrome who were treated in our department over the last 19 years were analyzed retrospectively. Removal of adenoma by the transsphenoidal approach was done in 21 patients and by transfrontal craniotomy in 2. The follow-up period ranged from six months to nine years. RESULTS: The incidence of Nelson's syndrome was 7.7% in a series of 300 patients with Cushing's disease treated by microsurgery in the same period. Hyperpigmentation was relieved and adrenocorticotropic hormone (ACTH) levels decreased in all patients after tumor excision. Eight patients with visual disturbance improved after surgery. The curative and remission rates were 56.5% and 26.1%, respectively. CONCLUSIONS: Transsphenoidal microsurgical removal of pituitary ACTH adenoma is the first choice in the prevention and treatment of Nelson's syndrome. Regular follow-up examinations should be performed over a long time.

[A novel gene involved in blood glucose regulation].

OBJECTIVE: To clone a novel gene relative to blood glucose regulation. METHODS: Rat modes of autonomous regulation of blood glucose was made by intra jugular vein to right atrium injection of high concentration of glucose solution, and the control rats were injected with 0.9%NaCl both before skeletal muscles were separated for gene analysis. The differentially expressed fragments were identified by differential display technology (DDRT-PCR). After slot blot and Northern blot analysis, the artificial positive fragments were excluded and the true EST (expression sequence tag) differentially expressed was obtained. These positive EST were used as probes to screen cDNA library of rat skeletal muscle. RESULTS: A novel full-length cDNA, named as Fang-2 was obtained. GenBank Accession No. was AF399874. Fang-2 was found rat homologue of human troponin T by blast software (NCBI). It shared 78% identical nucleotides, which showed the family proteins were conservative. After high concentration of glucose stimulation of rats, the expression of Fang-2 was down-regulated. CONCLUSIONS: A novel gene relative to blood glucose regulation was cloned from rat skeletal muscle. The gene can regulate blood glucose level by effect certain mechanisms unknown yet with down-regulation expression.

[Serial analysis of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues].

OBJECTIVE: To observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues. METHODS: Using serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI. RESULTS: A total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively. CONCLUSIONS: Genes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes.