Down-regulation of microRNA-126-5p contributes to overexpression of VEGFA in lipopolysaccharide-induced acute lung injury.

OBJECTIVES: To investigate the role of microRNA-126-5p (miR-126-5p) in acute lung injury induced by bronchial instillation of lipopolysaccharide (LPS), and to explore the potential target(s) of miR-126-5p in acute lung injury. RESULTS: In the mice with LPS-induced acute lung injury, the level of miR-126-5p in the pulmonary tissues was decreased by 41 % whilst pulmonary vascular endothelial growth factor-A (VEGFA) doubled in its mRNA content and increased threefold in its protein level. Similar results were observed in the alveolar type II (ATII) cells treated with LPS. By using luciferase reporter assay, we found that miR-126-5p inhibited VEGFA expression by targeting its 3'-untranslated region. In addition, overexpression of miR-126-5p attenuated LPS-induced reduction of epithelial sodium channel and aquaporin 1 in ATII cells CONCLUSIONS: MiR-126-5p was down-regulated in LPS-induced acute lung injury in mice. Thus overexpression of miR-126-5p may alleviate acute lung injury by down-regulating VEGFA.

Comparison of burst and tonic spinal cord stimulation on spinal neural processing in an animal model.

OBJECTIVES: Spinal cord stimulation (SCS) using bursts of pulses suppressed neuropathic pain as well or better than tonic stimulation and limited the incidences of parasthesias. The present translational study explored possible differences in mechanisms of burst and tonic SCS on nociceptive spinal networks and/or the gracile nucleus supraspinal relay. MATERIALS AND METHODS: Visceromotor reflexes (VMRs, a nociceptive response) or extracellular activity of either L6-S2 spinal neurons or gracile nucleus neurons were recorded during noxious somatic stimulation (pinching) and visceral stimulation (colorectal distension [CRD]) in anesthetized rats. A stimulating (unipolar, ball) electrode at L2-L3 delivered 40 Hz burst or tonic SCS at different intensities relative to motor threshold (MT). RESULTS: Average MTs for burst SCS were significantly lower than for tonic SCS. Burst SCS reduced the VMR more than tonic SCS. After high-intensity SCS (90% MT), spinal neuronal responses to CRD and pinch were reduced similarly for burst and tonic SCS. At low-intensity SCS (60% MT), only burst SCS significantly decreased the nociceptive somatic response. Tonic but not burst SCS significantly increased spontaneous activity of neurons in the gracile nucleus. CONCLUSION: Based on the clinically relevant burst versus tonic parameters used in this study, burst SCS is more efficacious than tonic SCS in attenuating visceral nociception. Burst and tonic SCS also suppress lumbosacral neuronal responses to noxious somatic and visceral stimuli; however, burst SCS has a greater inhibitory effect on the neuronal response to noxious somatic stimuli than to noxious visceral stimuli. Reduced or abolished paresthesia in patients may be due in part to burst SCS not increasing spontaneous activity of neurons in the gracile nucleus.

Is constant current or constant voltage spinal cord stimulation superior for the suppression of nociceptive visceral and somatic stimuli? A rat model.

OBJECTIVES: This study compares the effects of constant current (CC) and constant voltage (CV) spinal cord stimulation (SCS) at various frequencies and intensities on standard nociceptive measurements in rats, the visceromotor reflex (VMR) and neuronal activity, during noxious visceral and somatic stimuli. MATERIALS AND METHODS: Abdominal muscle electromyographic activity changes were measured to indicate VMR, and extracellular activity of L6-S2 spinal neurons was recorded during somatic (pinching) and noxious visceral stimulation (colorectal distension [CRD], 60 mmHg) in anesthetized rats. A stimulating (unipolar ball) electrode at L2-L3 delivered CC- or CV-SCS at varied frequencies and intensities. RESULTS: CC-SCS reduced VMR evoked by CRD significantly more than CV-SCS (p < 0.05). For neuronal activity, high-frequency CC-SCS (40 and 100 Hz) and CV-SCS (100 Hz) effectively reduced intraspinal somatic nociceptive transmission more than low-frequency SCS (2 Hz). No significant differences were observed between the effects of CC- and CV-SCS on spontaneous activity and nociceptive responses of spinal neurons to noxious CRD following short- (five to ten minutes) or long-term (20-30 min) SCS. CONCLUSIONS: Although high-frequency CC- and CV-SCS may be more useful for the management of somatic pain, CC-SCS may be more effective for treating complex pain systems like visceral hypersensitivity.

Methylation of SPRED1: A New Target in Acute Myeloid Leukemia.

Sprouty-related, EVH1 domain-containing protein 1 (SPRED1) has been identified as a novel tumor suppressor gene in acute myeloid leukemia (AML). Previous studies showed that SPRED1 methylation levels were significantly increased in AML patients, making it an interesting candidate for further investigations. To confirm the association of SPRED1 methylation, clinical parameters, and known molecular prognosticators and to identify the impact of methylation level on treatment outcome, we conducted this study in a larger cohort of 75 AML patients. Significantly increased methylation levels of SPRED1 were detected at four of ten CpG units by quantitative high-resolution mass spectrometry-based approach (MassARRAY) in AML patients. Whereas overall survival (OS) and relapse-free survival (RFS) showed no statistical difference between hypermethylation and hypomethylation subgroups, the relationship between methylation level and treatment response was indicated in paired samples from pre- and post-induction. To determine the possible mechanism of SPRED1 methylation in AML, we performed in vitro experiments using THP-1 cells, as the latter showed the highest methylation level (determined by utilizing bisulfite modification) among the three AML cell lines we tested. When treated with 5-AZA and lentivirus transfection, upregulated SPRED1 expression, decreased cell proliferation, increased cell differentiation and apoptosis, and inactivated phosphorylated extracellular signal-regulated kinase (p-ERK) were detected in THP-1 cells. These results show that demethylation of SPRED1 can inhibit the proliferation of AML cells and promote their differentiation and apoptosis, possibly by the ERK pathway. The hypermethylation of SPRED1 is a potential therapeutic target for AML.

Ketamine and propofol in combination induce neuroapoptosis and down-regulate the expression of N-methyl-D-aspartate glutamate receptor NR2B subunit in rat forebrain culture.

BACKGROUND AND OBJECTIVE: Ketamine has always been used in combination with propofol in paediatric patients. Ketamine interacts with N-methyl-D-aspartate glutamate receptor to exert its biologic actions. The NMDA receptor NR2B subunit is expressed at nearly adult level during forebrain development in the cortex and considered as the major type of functional NMDA receptor in the central nervous system. This study aimed to investigate whether ketamine or ketamine in combination with propofol induces apoptosis and regulates the expression level of NMDA receptor NR2B subunit in rat forebrain culture. METHODS: Rat primary forebrain cultures were exposed to different concentrations of ketamine (1 microM, 10 microM, 20 microM) or 20 microM ketamine plus 5 microM propofol on the 6th day for 12 h. Cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Caspase-3 activity was measured and apoptotic morphology was examined by Hoechst dye staining. The expression of NMDA receptor NR2B subunit at mRNA and protein level was determined by qPCR and Western blot, respectively. RESULTS: Ketamine (10 microM and 20 microM) or 20 microM ketamine plus 5 microM propofol resulted in a significant decrease in cell viability, and a significant increase in caspase-3 activity and apoptosis of primary rat forebrain culture. The expression of NMDA receptor NR2B subunit at both mRNA and protein levels were down-regulated by administration of ketamine (1 microM, 10 microM and 20 microM) and 20 microM ketamine plus 5 microM propofol. CONCLUSION: Ketamine (10 microM or 20 microM) alone or in combination with propofol (20 microM ketamine plus 5 microM propofol) induces neuroapoptosis and down-regulates of NMDA receptor NR2B subunit in rat forebrain culture.

Autologous transplantation of adipose-derived stromal cells combined with sevoflurane ameliorates acute lung injury induced by cecal ligation and puncture in rats.

Adipose-derived stromal cells (ADSCs) have excellent capacities for regeneration and tissue protection, while sevoflurane, as a requisite component of surgical procedures, has shown therapeutic benefit in animal models of sepsis. This study therefore determined if the combination of sevoflurane and ADSCs exerted additional protective effects against acute lung injury (ALI) induced by cecal ligation and puncture (CLP) in rats. The animals were randomized into five groups: (sham operation (group I), CLP followed by mechanical ventilation (group II), CLP plus sevoflurane at 0.5 minimum alveolar concentration (group III), CLP plus intravenous autologous 5 × 106 ADSCs (group IV), and CLP plus sevoflurane and ADSCs (group V). Levels of the pro-inflammatory cytokines tumor necrosis factor-α, transforming growth factor-ß1, interleukin-1ß and interleukin-6 were significantly increased in CLP rats. Moreover, epithelial sodium channel expression levels and activities of Na/K-ATPase and alveolar fluid clearance were significantly reduced in CLP-induced ALI rats. ADSCs improved all these parameters, and these effects were further enhanced by the addition of sevoflurane. In conclusion, combined treatment with ADSCs and sevoflurane is superior to either ADSCs or sevoflurane therapy alone for preventing ALI. This beneficial effect may be partly due to improved alveolar fluid clearance by the paracrine or systemic production of keratinocyte growth factor and via anti-inflammatory properties.

Large Tracheocutaneous Fistula Successfully Treated With Bronchoscopic Intervention and Flap Grafting: A Case Report and Literature Review.

Tracheocutaneous fistula (TCF) is the most common related post-operative complication after tracheotomy. Treatments such as surgery and flap grafting are usually applied to close TCFs. We report a case of a large TCF with an area of ~3.0 cm × 1.0 cm. Here, we describe a relatively convenient approach for the management of a patient with a large TCF. In our treatment strategy, a coverd tracheal stent was used to cover the defect by bronchoscopy, the bronchial defect was closed with a local turnover flap, the structure was reinforced with biodegradable material (RapidSorb Plate 2.0), and then transplantation of a deltopectoral flap was performed. It is worth noting that the patency of the trachea was maintained during the whole surgery course. No recurrence or complications occurred after the 12-month follow-up. The large TCF was successfully treated with bronchoscopic intervention, biodegradable material and flap grafting, and without cartilage grafting.

SPRED1 Is Downregulated and a Prognostic Biomarker in Adult Acute Myeloid Leukemia.

We report herein that Sprouty-Related EVH1 Domain-Containing Protein1 (SPRED1) is downregulated and a prognostic biomarker in adult acute myeloid leukemia (AML). We determined mRNA levels of SPRED1 in the bone marrow mononuclear cells from adult patients, including 113 AMLs and 22 acute lymphoblastic leukemias (ALLs), as well as in 37 healthy control subjects. Significantly decreased SPRED1 mRNA expression was found in AML patients comparing to those in ALL patients and healthy controls, which was confirmed by immunocytochemistry analysis of SPRED1 protein and ELISA measurement of serum SPRED1 level. Further analysis demonstrated that SPRED1 expression was significantly higher for most patients at complete remission after induction treatment than at diagnosis. Moreover, SPRED1 expression was significantly downregulated in M2 and M3 types. Non-acute promyelocytic leukemia (non-APL) patients with decreased SPRED1 had significantly lower 2-year progression-free survival and event-free survival rates. In vitro, ectopic overexpression of SPRED1 leads to a decrease of extracellular signal-regulated kinase (ERK) phosphorylation, induction of apoptosis and reduction of proliferation of THP-1 cells. Our findings suggest SPRED1 is not only a predictor of treatment response, but also an independent prognostic factor for non-APL, and targeting Ras- Mitogen-activated protein kinase (MAPK) signaling may be a promising strategy for the treatment of AML with downregulation of SPRED1.

Multiple extramedullary plasmacytomas of the trachea and pharyngeal tissue: a case report and literature review.

Extramedullary plasmacytoma (EMP) is an infrequent form of plasma cell dyscrasia that presents as a mass of monoclonal plasma cells located in extramedullary soft tissues with no skeletal component. EMP constitutes ~4% of all plasma cell neoplasms and occurs mostly in the upper respiratory tract. Here, we report a rare case of multiple EMPs involving the trachea, laryngopharynx, and oropharynx, which caused symptoms of airway obstruction as the only clinical manifestation. The patient was diagnosed by histopathology of the tissue from bronchoscopic resection and successfully managed with bronchoscopic intervention to treat lesions in the trachea and radiotherapy combined with surgical resection to treat lesions in the pharynx. There was no recurrence after 14 months of follow-up. Endoscopic intervention plays a key role in the rapid diagnosis and treatment of EMP involving the central airways.

Identification of novel genomic aberrations in AML-M5 in a level of array CGH.

To assess the possible existence of unbalanced chromosomal abnormalities and delineate the characterization of copy number alterations (CNAs) of acute myeloid leukemia-M5 (AML-M5), R-banding karyotype, oligonucelotide array CGH and FISH were performed in 24 patients with AML-M5. A total of 117 CNAs with size ranging from 0.004 to 146.263 Mb was recognized in 12 of 24 cases, involving all chromosomes other than chromosome 1, 4, X and Y. Cryptic CNAs with size less than 5 Mb accounted for 59.8% of all the CNAs. 12 recurrent chromosomal alterations were mapped. Seven out of them were described in the previous AML studies and five were new candidate AML-M5 associated CNAs, including gains of 3q26.2-qter and 13q31.3 as well as losses of 2q24.2, 8p12 and 14q32. Amplication of 3q26.2-qter was the sole large recurrent chromosomal anomaly and the pathogenic mechanism in AML-M5 was possibly different from the classical recurrent 3q21q26 abnormality in AML. As a tumor suppressor gene, FOXN3, was singled out from the small recurrent CNA of 14q32, however, it is proved that deletion of FOXN3 is a common marker of myeloid leukemia rather than a specific marker for AML-M5 subtype. Moreover, the concurrent amplication of MLL and deletion of CDKN2A were noted and it might be associated with AML-M5. The number of CNA did not show a significant association with clinico-biological parameters and CR number of the 22 patients received chemotherapy. This study provided the evidence that array CGH served as a complementary platform for routine cytogenetic analysis to identify those cryptic alterations in the patients with AML-M5. As a subtype of AML, AML-M5 carries both common recurrent CNAs and unique CNAs, which may harbor novel oncogenes or tumor suppressor genes. Clarifying the role of these genes will contribute to the understanding of leukemogenic network of AML-M5.