Enterocolitis, Necrotizing ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Infant, Newborn ; Intestinal Mucosa ; metabolism ; pathology ; Male ; Platelet Activating Factor ; metabolism

BACKGROUNDReactive oxygen species (ROS) may play both physiological and pathophysiological roles. Transcription factor NF-E2-related factor 2 (Nrf2) regulates antioxidant response element (ARE)-mediated genes expression and coordinates induction of chemoprotective proteins in response to physical and chemical stresses. The exact role of Nrf2 in cellular responses to different levels of oxidative stresses remains unknown.

METHODSRat pulmonary microvascular endothelial cells were cultured and treated with 0 mmol/L, 0.125 mmol/L, 0.25 mmol/L, 0.5 mmol/L, 1.0 mmol/L and 2.0 mmol/L hydrogen peroxide solution for 2 hours. Nrf2 gene expression was assayed by reverse transcription-PCR, Nrf2-ARE binding activity was assayed with electrophoretic mobility shift assay (EMSA), and localization of Nrf2 was detected with immunohistochemistry.

RESULTSLow and moderate (0.125 mmol/L, 0.25 mmol/L and 0.5 mmol/L) doses hydrogen peroxide exposure of rat pulmonary microvascular endothelial cells led to the nuclear accumulation of Nrf2, increased activity of transcription regulation and up-regulation of ARE-medicated gene expression. In contrast, high doses of hydrogen peroxide (1 mmol/L, 2 mmol/L) exposure of the cells led to the nuclear exclusion of Nrf2, decreased activity transcription regulation and down-regulation of ARE-mediated gene expression.

CONCLUSIONLow and moderate doses of hydrogen peroxide play protective roles by increasing transcription activity of Nrf2, whereas high- dose hydrogen peroxide plays a deleterious role by decreasing transcription activity of Nrf2.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cells, Cultured ; Electrophoretic Mobility Shift Assay ; Gene Expression ; drug effects ; Hydrogen Peroxide ; pharmacology ; Immunohistochemistry ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

This study was aimed to investigate whether endothelium-specific deletion of PTEN can affect hemangioblast development in the AGM region of mouse embryos. Based on Cre/loxP system, the Tie2CrePten(loxp/loxp) and Tie2CrePten(loxp/wt) mouse embryos were obtained. The genotype was identified by PCR. After treated with type I collagenase, the AGM region was dispersed into single-cell suspension, and then was cultured in blast colony-forming cell (BL-CFC) media. The number of BL-CFC was counted 4 or 5 days later. The hematopoietic capacity of BL-CFC was detected in methylcellulose culture system and the endothelial potential was assessed by tube-like structure formation on Matrigel. The results showed that the number of BL-CFC in AGM region of Tie2CrePten(loxp/loxp) mouse embryo decreased as compared with Tie2CrePten(loxp/wt) embryo. Whereas the hematopoietic capacity of mutant BL-CFC was enhanced, the endothelial potential, as evaluated by tube-like structure formation in vitro, was significantly reduced. It is concluded that the endothelial PTEN is capable of exerting regulatory functions on both the numbers and the dual potential of hemangioblast in mouse AGM region.
Animals ; Cell Differentiation ; Cells, Cultured ; Hemangioblasts ; Hematopoietic Stem Cells ; cytology ; Mice ; PTEN Phosphohydrolase ; genetics

OBJECTIVETo explore the association between chromosomal disequilibrium and chemoresistance/chemosensitivity in non-small cell lung cancer (NSCLC) using comparative genomic hybridization (CGH).

METHODSGenomic DNA samples were prepared from the tumor tissues in paraffin-embedded sections derived from 88 patients with advanced NSCLC (18 with chemosensitivity and 16 with chemoresistance). The DNAs were first amplified by a degenerate oligonucleotide prime-polymerase chain reaction protocol and then labeled with fluorescence as probes for CGH analyses. The correlations of the resulting chromosomal imbalances with the chemo-sensitivity and other pathological features of the patients were analyzed.

RESULTSA total of 640 abnormal chromosome regions including 96.12% gains and 3.88% losses were detected in 88 specimens. The results indicated that the most frequently gained chromosome regions were 19p13.1-13.3 (39/88, 44.12%), followed by 9q12-q22 (26/88, 29.41%), 22q12-q13 (26/88, 29.41%), and Xq (29/88, 32.35%). The total number of abnormal regions related with chemo-sensitivity was 188( 182 gains and 6 losses), while the number of the abnormal regions linked to the chemoresistance was 452 (431 gains and 21 losses) (P=0.005). Gains of 14p12-p13 and 19p were significantly correlated with the chemosensitivity of the NSCLC (P=0.006). Gains of 1q12-q22, 10q25-q26, 5p15.1-p15.3, 19q13.2-13.4, 20p11.2-p12, 21q22, and Xp 21-p22.1 were also significantly correlated with the chemoresistance (P]0.005, 0.029, 0.039, 0.029, 0.039, 0.016, and 0.006, respectively). No correlation between the chromosome abnormalities and other clinical features was observed.

CONCLUSIONSThe specific gains and losses of chromosome region is correlated with platinum-based first-line chemotherapy in NSCLC patients,as confirmed by CGH detection. This finding is useful for further identifying the chemosensitivity-related functional genes, predicting clinical effectiveness, and achieve individualized treatment in the future.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Chromosome Aberrations ; Comparative Genomic Hybridization ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Karyotyping ; Lung Neoplasms ; drug therapy ; genetics ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo study the diagnostic value of determinations of serum acute phase reaction protein, such as complement 3 (C3), complement 4 (C4), prealbumin (PA) and C-reactive protein (CRP), etc., in patients with severe acute respiratory syndrome (SARS).

METHODSSerum levels of C3, C4, PA and CRP were determined by turbidimetry in 54 cases of SARS, 20 of other pneumonia and 30 normal persons.

RESULTSSerum concentrations of C3, C4, CRP and PA were (1.18 +/- 0.42) g/L, (1.15 +/- 0.56) g/L, (10.52 +/- 8.77) mg/L and (107 +/- 54) mg/L in SARS patients, (1.30 +/- 0.46) g/L, (0.57 +/- 0.31) g/L, (0.88 +/- 0.43) mg/L and (291 +/- 76) mg/L in patients with other pneumonia, and (1.11 +/- 0.56) g/L, (0.38 +/- 0.26) g/L, (0.42 +/- 0.26) mg/L and (376 +/- 74) mg/L in normal persons, respectively. Serum level of PA was significantly lower and levels of C4 and CRP significantly in patients with SARS higher than those in patients with other pneumonia and normal persons (P < 0.01). There was no significant difference in serum level of C3 between the three groups (P > 0.05).

CONCLUSIONDetermination of serum level of C4, CRP and PA in suspected patients is beneficial to early differential diagnosis for SARS.

Acute-Phase Proteins ; analysis ; Acute-Phase Reaction ; blood ; Adult ; Aged ; C-Reactive Protein ; analysis ; Complement C3 ; analysis ; Complement C4 ; analysis ; Female ; Humans ; Male ; Middle Aged ; Prealbumin ; analysis ; Severe Acute Respiratory Syndrome ; blood

The fusion protein (F) gene of Newcastle disease virus was amplified by polymerase chain reaction (PCR) from the recombinant plasmid pVAX1-F, and subcloned into eukaryotic expression vector pmcDNA3. 1+. The F gene was identified by sequencing. The recombinant plasmid was transformed into attenuated Salmonella typhimurium SL7207, and the recombinant was designated as SL7207 (pmcDNA3. 1-F). In vitro and in vivo experiments showed that the plasmid stability of pmcDNA3. 1-F was apparently higher than that of pcDNA3. 1-F in SL7207. In order to compare the immune response induced by these two re combinant bacteria, BALB/c mice were immunized orally with them at the dosage of 2 x 10(9) CFU respectively. Both SL7207(pcDNA3. 1-F) and SL7207(pmcDNA3. 1-F) initiated F-specific serum and mucosal antibodies in immunized mice. Furthermore, 4-day-old SPF chickens were immunized with SL7207(pcDNA3. 1-F) and SL7207(pmcDNA3. 1-F) at the dosage of 5 x 10(9) CFU and boosted two weeks later with the same dosage. Humoral and intestinal mucosal immune responses were observed and their levels were significantly higher than that of negative and positive controls. The result of protective efficacy showed that the chickens immunized with SL7207(pmcDNA3. 1-F) had the protective rate of 70.0%, higher than that of the SL7207 (pcDNA3. 1-F) with 50.0%. In summary, the DNA vaccine delivered by attenuated Salmonella typhimurium has good immunogenicity. A novel mucosal DNA vaccine has been developed and could be useful for controlling the infection and epidemic of Newcastle disease in the poultry.
Animals ; Chickens ; Female ; Immunization ; Mice ; Mice, Inbred BALB C ; Newcastle disease virus ; immunology ; Plasmids ; Salmonella typhimurium ; genetics ; Vaccines, Attenuated ; immunology ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

BACKGROUNDIbutilide has been commonly used for pharmacologic cardioversion of atrial fibrillation and flutter in clinical settings. The objective of this study was to investigate the effects of ibutilide on the defibrillation threshold (DFT), restitution properties, dispersion of refractoriness and activation patterns during ventricular fibrillation (VF).

METHODSIbutilide was administrated intravenously in six open-chest beagles. Before and after the drug administration, 20-second episodes of VF were electrically induced and recorded with a 10×10 unipolar electrode plaque sutured on the lateral epicardium of the left ventricle. DFT and VF activation patterns, including type of epicardial activation maps, VF cycle length (VF-CL), conduction velocity, wavelength (WL) and reentry incidence, were measured. Restitution properties and dispersion of refractoriness were estimated from activation recovery intervals (ARI) during pacing.

RESULTSCompared to baseline, ibutilide markedly decreased the DFT by 31% ((491 ± 14) V vs. (337 ± 59) V, P < 0.01). The drug significantly reduced the maximal slope of the restitution curve (1.34 ± 0.08 vs. 0.76 ± 0.06, P < 0.01) and its epicardial dispersion (0.36 ± 0.09 vs. 0.21 ± 0.06, coefficient of variation, P = 0.03). The dispersion of refractoriness was enhanced at the pacing cycle length of 300 ms to 160 ms by ibutilide. The drug significantly increased the VF-CL ((96 ± 19) ms vs. (112 ± 20) ms, P < 0.01) and the WL ((41 ± 9) mm vs. (52 ± 14) mm, P = 0.02) during VF, and reduced the reentry incidence by 25% (0.08 ± 0.02 vs. 0.06 ± 0.02, P < 0.01). In the epicardial activation maps, ibutilide significantly reduced the percentage of more complex activation maps during VF.

CONCLUSIONSIntravenous ibutilide significantly decreased the DFT. It might be due to reduction of activation pattern complexity during VF.

Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Dogs ; Pericardium ; drug effects ; Sulfonamides ; therapeutic use ; Ventricular Fibrillation ; drug therapy ; physiopathology

This study was purposed to investigate the effect of RUNX1 on transcription activity of WNT5A promoter in mouse bone marrow derived mesenchymal stem cells (MSC), and to explore the mechanism by which bone marrow environments regulate MSC. RT-PCR was used to detect the expression of RUNX1 in MSC isolated from mouse bone marrow and cultured in vitro; the chromatin immunoprecipitation (ChIP) was used to investigate the direct in vivo interaction between the RUNX1 and WNT5A promoter; retrovirus system was utilized to introduce the RUNX1 gene into MSC to detect the regulation of RUNX1 on the transcription activity of WNT5A promoter. The results showed that mouse bone marrow derived MSC was positive for Oil Red O, van Kossa and toluidine blue staining respectively and RUNX1 expressed in MSC. WNT5A promoter could be bound by RUNX1, and the expression level of WNT5A was enhanced with the increase of RUNX1. It is concluded that RUNX1 expresses in mouse bone marrow derived MSC, WNT5A is a direct target gene of RUNX1 and its transcriptional activity is regulated by RUNX1.
Animals ; Bone Marrow Cells ; metabolism ; Cell Differentiation ; Cells, Cultured ; Chromatin Immunoprecipitation ; Core Binding Factor Alpha 2 Subunit ; genetics ; Mesenchymal Stromal Cells ; metabolism ; Mice ; Mice, Inbred C57BL ; Transcription, Genetic ; Wnt Proteins ; genetics ; Wnt-5a Protein

Ischemia-reperfusion injury (IRI) is a major complication in liver transplantation (LT) and it is closely related to the recovery of grafts' function. Researches has verified that both innate and adaptive immune system are involved in the development of IRI and Kupffer cell (KC), the resident macrophages in the liver, play a pivotal role both in triggering and sustaining the sterile inflammation. Damage-associated molecular patterns (DAMPs), released by the initial dead cell because of the ischemia insult, firstly activate the KC through pattern recognition receptors (PRRs) such as toll-like receptors. Activated KCs is the dominant players in the IRI as it can secret various pro-inflammatory cytokines to exacerbate the injury and recruit other types of immune cells from the circulation. On the other hand, KCs can also serve in a contrary way to ameliorate IRI by upregulating the anti-inflammatory factors. Moreover, new standpoint has been put forward that KCs and macrophages from the circulation may function in different way to influence the inflammation. Managements towards KCs are expected to be the effective way to improve the IRI.
Cytokines ; Hand ; Immune System ; Inflammation ; Ischemia* ; Kupffer Cells ; Liver Transplantation* ; Liver* ; Macrophages* ; Receptors, Pattern Recognition ; Reperfusion Injury* ; Reperfusion* ; Toll-Like Receptors

Objective To evaluate the incidence, prognosis and influencing factors of lower cranial nerve palsy after microvascular decompression (MVD) in patients with hemifacial spasm (HFS). Methods Clinical data of 1033 patients with HFS, admitted to our hospital from May 2014 to August 2016, were retrospectively analyzed. The incidence of lower cranial nerve palsy after procedure and prognosis of these patients were summarized; and influencing factors were analyzed by statistical method. Results In 1033 patients with HFS, 10 patients (0.97％) had lower cranial nerve palsy after procedure: 6 patients suffered hoarseness, 2 patients suffered dysphasia, and 2 patients were with both symptoms; the symptoms in 6 patients completely recovered within one month, 2 patients within one-3 months, and 2 patients within 3-6 months. No permanent low cranial nerve palsy occurred. There were no statistical significances in age, gender, clinical course, sides, or the types of offending vessels between the lower cranial nerve palsy patients (n=10) and the non-palsy patients (n=1023)(P>0.05). Conclusion Lower cranial nerve palsy is a rare complication after MVD, and type of responsible vessels is not a influencing factor in this complication